CNR Environment and Health Inter-departmental Project: present knowledge and prospects for future research

Consiglio Nazionale delle Ricerche
Dipartimento Terra e Ambiente
CNR Environment and Health
Inter-departmental Project: present
knowledge and prospects for future
CNR Environment and Health Inter-departmental Project
Fabrizio Bianchi, Liliana Cori, Pier Francesco Moretti
Editorial Office
Anna Russo, Tiziana Siciliano
Composition and Copy Editing
Luigi Mazari Villanova
Web release
Daniela Beatrici (
Print: Edizioni Nuova Cultura
Publisher: Consiglio Nazionale delle Ricerche - Roma
© 2010, Consiglio Nazionale delle Ricerche
All rights reserved
ISBN 978-88-8080-113-9
Cover image courtesy of Alfonso Mascia, “Flamingo Nursery at Saline
Contivecchi”, Stagno di Santa Gilla, Cagliari, Italy
PIAS logo designer: Luca Duè, Pisa, Italy
The National Research Council (CNR) is the major public research organisation in Italy;
its duty is to carry out, promote, spread, transfer and improve research activities in the
main sectors of knowledge growth and of its applications for the scientific, technological,
economic and social development of the Country.
To this end, CNR activities are organised into macro areas of interdisciplinary scientific
and technological research covering several sectors: biotechnology, medicine, materials,
environment andthe territory, information and communications, advanced production
systems, judicial and socio-economic sciences, classical studies and arts.
Increasing knowledge about the complex interaction between environment and health is
a major objective and is recognized as a worldwide challenge that the Italian National
Research Council (CNR) wishes to take up.
Our effort aims at transferring the knowledge developed by our research institutes on
environment and health issues to our society, providing local, national and international
governments with priorities to be applied in social policies.
In the EH sector, CNR carries out institutional R&D activities in coordination with
the Ministry of University and Research and, apart from these institutional activities,
CNR and the Italian Ministry of the Environment have signed numerous agreements
and contracts of project implementation, consultancy and service activities. CNR is also
involved in programmes and projects of the Ministry of Health.
The Department of Earth and Environment (DTA) includes thirteen research Institutes
involved in research, surveys, technologies and knowledge transfer in a wide range of
Earth sciences addressing atmospheric, terrestrial and aquatic issues. The mission of DTA
is to gather knowledge and predict the behaviour of the earth system and its resources to
contribute to a sustainable future for the planet and mankind.
The Department of Medicine includes twelve research Institutes performing basic,
clinical and epidemiological research on many types of diseases as well as on disease
prevention also through the interaction with National Health System structures. Tasks of
the Department of Medicine include also innovation and technology transfer in medicine
as well as education and training of personnel on health-related themes.
A total of thirty-seven Institutes, including some belonging at the Departments of
Agriculture and Food, Materials and Devices, Energy and Transportation and ICT, are
involved in research on EH interface.
In 2006 the CNR started a multi-disciplinary program aiming at promoting and coordinate
collaborative research and joint actions on environment and health. In mid 2007, following
priorities set also by the European Union, the CNR activated an Inter-departmental Project
called Environment and Health (Italian acronym: PIAS CNR).
CNR Environment and Health Inter-departmental Project
The Environment and Health Project has the overall objective of promoting an integrated
research in these two scientific sectors, and the specific objective of developing the
knowledge on sources of pollution and the consequent negative effects on health, the
instruments and methods to analyse the interaction between environment and health, the
instruments and methods to be used in managing complex situations.
As operative instruments, activities were identified in collaboration with CNR researchers,
to strengthen competences and to facilitate the participation in international networks.
CNR is working to promote and transfer the results of research to the production system
and to decision makers; to promote the experience of Health Impact Assessment on
programmes, plans and policies; to contribute to developing Environment and Health
monitoring systems; to test and validate environment and health indicators suggested by
the EU and the WHO, supported by suitable information systems; to test and validate the
use of geographic Information System (GIS) for a joint evaluation of territorial health and
environmental data and to improve instruments and methods for risk reporting through
recommendations and guidelines.
The PIAS CNR project is based upon the multidisciplinary activity of researchers of CNR
and other public bodies, including the WHO, the Italian National Institute of Health,
Italian Universities, the National Health System, the Ministry of the Environment and
Territory, the Ministry of Health, the Regional Agency for Environmental Protection and
Epidemiological Observatories.
The present publication includes overviews, results and suggestions produced by the
PIAS working groups on effect of pollutants in the soil, water and soil monitoring for
the protection of the environment and human health, the role of atmospheric pollution in
terms of its harmful effects on health, Human bio-monitoring, Environment and Health
surveillance systems, Monitoring contaminants in the food chain and their impact on
human health. Two Pilot Studies on “Endocrine disruptors and their effects on health”
and “Ultrafine particles and cardiopulmonary effects” are in progress.
A PIAS conclusive international workshop is planned for late 2010; this will represent
a chance for the CNR Institutes as well as for collaborative Institutions for sharing
experiences and conclusions between research groups and plan future research.
Giuseppe Cavarretta
& Gianluigi Condorelli
CNR Department of
Earth and Environment
CNR Department of Medicine
Fabrizio Bianchi, Liliana Cori, Pier Francesco Moretti
The fate of pollutants in soil
G. Petruzzelli, F. Gorini, B. Pezzarossa, F. Pedron
1. Introduction
2. State of the art and interactions between basic and applied research
3. Pilot Study: Influence of soil characteristics on the mobility of contaminants in the
industrial area of Gela (Sicily)
4. CNR Specific expertise: qualified teams and external collaborations
5. Future perspectives and developments
Water and Soil Monitoring for the Protection of Environment and
Human Health
M. Rusconi, S. Polesello
1. Introduction
2. Working methodology in working group 2 of the CNR Environment and Health InterDepartmental Project (PIAS)
3. State of the art of CNR activities
4. Emerging issues
5. Future perspectives and developments
Role of Atmospheric Pollution on Harmful Health Effects
A. Pietrodangelo, M. Bencardino, A. Cecinato, S. Decesari, C. Perrino, F. Sprovieri,
N. Pirrone and M.C. Facchini
1. Introduction
2. Air Pollution and Health
3. Inorganic air pollutants
4. Organic air pollutants
5. Particles in the ultrafine (UFP) and nano-size fractions
6. Future perspectives and developments in the framework of the “pilot study for the
assesment of health effects of the chemical composition of ultrafine and fine particles in
Italy” project
Human Biomonitoring
E. Leoni, A. I. Scovassi
CNR Environment and Health Inter-departmental Project
1. Introduction
2. From basic to applied research
3. The CNR specific expertise: qualified teams and external collaborations
4. Relevant findings
5. Future perspectives and developments
Environmental Health Surveillance Systems
E.A.L. Gianicolo, A. Bruni, M. Serinelli
1. Introduction
2. Environmental and health surveillance: state of the art
3. Environmental and health surveillance experiences
4. Epidemiological and environmental characterization of Brindisi (experimental site)
5. Proposal for an enviromental health surveillance system
6. Conclusions
Monitoring Contaminants in Food Chain and their Impact on Human
A. Mupo, F. Boscaino, G. Cavazzini, A. Giaretta, V. Longo, P. Russo, A. Siani, R.
Siciliano, I. Tedesco, E. Tosti, G.L. Russo
1. Background
2. State of the art and interaction between basic and applied research
2.1 Food quality control
3. CNR specific expertise: qualified teams, external collaborations and funding
4. Highlights
5. Future Perspectives: the effects of low-dose exposure to dietary contaminants in
children and young adults: a working hypothesis
6. Conclusion
The pilot study on Endocrine Disruptors
D.G. Mita
1 Introduction
2. State of the art: international and national initiatives
3. Ground and content of the pilot study
4. Expected results
Pilot study for the assessment of health effects of the chemical composition of ultrafine and fine particles in Italy
M.C. Facchini, F. Cibella, S. Baldacci, F. Sprovieri
1. Background
2. Objectives
CNR Environment and Health Inter-departmental Project:
present knowledge and prospects for future research
The World Health Organization states that the environment influences our health in many
ways through the exposure to physical, chemical and biological risks, and through the
changes that our behavior operates in response to those factors. This is what is commonly
addressed as the “Environment and Health issue” (EH), which has gained an increasing
attention at local, national and international level.
There is a growing political support to the implementation of concrete actions, as well as an
ever increasing knowledge of these issues. The need has been recognized to jointly tackle
these issues with the aim of producing more detailed and focused studies and planning
actions to reduce the negative impacts of some human activities on the environment.
Several international initiatives addressed this issue in the last years.
The environment and health issue in the European Union
Since the turn of the century, when the Lisbon Strategy was launched “to stimulate growth
and create more and better jobs, while making the economy greener and more innovative”,
the European Union has been playing a leading role in addressing the Environment
and Health issue. Following the Lisbon Strategy, the 2001 Gothenburg Council
established sustainable development as an overall objective for the EU development,
taking responsibility to ensure that future generations will not be forced to live in worse
conditions that the present one. An European Environment and Health Strategy has been
promoted in 2004, paving the way to the coordinated initiatives and joint efforts of the
enlarged European Union, and an Action Plan was produced to implement it.
In 2004, the European Commission issued the Environment and Health Action Plan
designed to combine information on environment, the ecosystem and human health so to
assess with greater effectiveness the overall impact of environment on human health.
The ultimate goal of the EU strategy is to set a cause-effect framework on environment
and health and provide the necessary information for the drafting of a EU policy on
pollution sources and the impact pathways of stress factors on health. The Action Plan
is based on some fundamental aspects: 1) to understand the relation between pollution
sources and health effects through the development of environment and health indicators
and integrated monitoring systems to assess human exposure and bio-monitoring
systems; 2) to enhance research and thus identify potential hazards to human health and
develop methodologies to analyse the environment-health relation; 3) to better inform the
population and improve risk communication systems.
In the European Union, several other initiatives have been active in the environment and
health sector: the Sixth Environmental Action Programme 2002-2010 contains specific
indicators measuring the reduction of health risks posed by the environment; the Public
Health Programme 2008-2013 adopts specific measures to reduce environment-related
health risks; the 6th and 7th Research and Development Framework Programme contains a
section on environment and health research initiatives. A number of other laws regarding
environment, energy production, infrastructures, territorial management, agriculture,
CNR Environment and Health Inter-departmental Project
have a direct impact on health: for this reason a unified Strategy and a surveillance system
is necessary and welcomed at the EU level.
The Environment and health issue for the United Nations and the World Health
One of the initiatives that can be considered as a milestone in the environment and health
sector is the list of the eight benchmarks provided by the United Nations Development
Programme in 2000, to be fulfilled within 2015: the Millennium Development Goals.(1)
All these Goals have in fact crucial implications for both the environment and health.
The setting up of a monitoring skill, with relevant indicators to be checked over the time
by each country Member of the UN, shall also be considered as an added value to this
On environment and health, however, the World Health Organisation (WHO) is the
leading international actor, whose paramount mission has always been to raise awareness
on health issues in all international Fora. In this context, it is important to point out
that the WHO uses a definition of the environment that is somewhat broader that the one
utilised by the European Commission in its Environment and Health Strategy; the WHO
definition includes socio-economic factors such as poverty and the lack of infrastructures,
whereas the Commission focuses on chemical and biological pollution. It seems desirable
that the EU new Constitutional Treaty could fill this gap and change the current scope of
its Articles 152 and 174.
In 1989, the WHO Office for Europe promoted the Environment and Health Process,
‘to raise awareness and start collaboration between sectors, particularly the health and
environment sectors’. The First Ministerial Conference on Environment and Health was
held in Frankfurt in 1989, gathering together the relevant public institutions of the 52
countries of the WHO European Region. The Second Conference was held in Helsinki in
1994, the Third in London in 1999.
The most recent Conference, held in Budapest in June 2004, launched the Children’s
Environment and Health Action Plan for Europe to improve the protection of future
generations. During this Conference, the European Commission presented the EU
Environment and Health Strategy and a specific Action Plan to implement it.
The Fourth Ministerial Conference on Environment and Health in Budapest 2004
The National Research Council of Italy (Italian acronym: CNR) was part of the Italian
delegation, led by the Ministry of the Environment, at the Budapest Conference. The
Italian representatives, in partnership with the Regional Environmental Centre for Central
and Eastern Europe (REC), proposed and organised awareness-raising initiatives - called
‘breathing days’ and focused on air pollution - in some Budapest schools, to underline the
active role that younger citizens could play in driving positive change.
CNR also participated in the workshop “Environment and Health in EU Structural Funds
Technical Assistance”, organised as a side event during the WHO Conference. The
workshop was a first opportunity to discuss the recent Italian developments with partners
from the large WHO-EU Region and to present all the coordination efforts made by Italy
to an international audience. Experiences developed in the 52 countries are undoubtedly
different and their analysis can be useful to plan future developments and actions.
The Authorities responsible of the use of the EU Structural Funds Technical Assistance
in Objective-1 Italian Regions (areas lagging behind in their development), gave to
the environment and health issue a relevance in the 2000-2006 planning period. CNR
worked together with the Ministries of the Environment and Health and the National
Health Institute, to establish an network of experts to analyse the specific issues of each
territory and address crucial environmental and health factors.
In Italy, the core competencies in environment and health are given to Regional
Authorities, and during the mentioned 2000-2006 planning period the European Union
allocated funds to the Ministry of the Environment for the establishment of Regional
Environmental Authorities to perform surveillance and monitoring activities; and to the
Ministry of Health to assist Regional Epidemiological Observatories.
In the framework of their technical assistance activities in seven Italian regions, the
Ministry of the Environment and the Ministry of Health joined their efforts to support
Regional Agencies for the Protection of the Environment and Regional Epidemiological
Observatories. The research on highly polluted sites has been selected as the first issue
to be developed. Polluted sites are in fact considered a priority in the agendas of regional
bodies and for the public opinion, and therefore these bodies had to be trained to develop
their environmental epidemiology competences, methodologies and tools. The technical
assistance of the two Ministries helped to implement activities and promoted a culture of
collaboration among the different regional bodies in charge of environment and health
protection, where the environmental epidemiology had the role of supplying a common
framework to experiment and develop such cooperation.
The Italian National Research Council (CNR)
The Italian National Research Council (CNR) competence on both the environment and
health is well established. The activities of the Department of Earth and Environment
(Italian acronym: DTA) are carried out by 13 research Institutes. DTA delivers independent
research, survey, technologies and knowledge transfer in the earth and environmental
sciences to advance knowledge on the planet as a complex and interacting system.
Research activities cover the full range of the Earth science, including atmospheric,
terrestrial and aquatic issues. The mission of DTA is to gather knowledge and predict the
behaviour of the earth system and its resources to help design a sustainable future for the
planet and mankind.
The Department of Medicine includes twelve institutes performing clinical and
epidemiological researches, both autonomously and by interacting with the National Health
System structures. Research and health activities include clinical and epidemiological
studies in cardiology, pneumology, oncology, neurology, immunology, infectious diseases,
genetics and molecular medicine. Innovation and technology transfer in medicine and
Education and the training of personnel and NHS are the other main activity areas.
The CNR report on areas presenting environmental hazards for health (2006)
In late 2006, the President of CNR and two Directors of Departments were asked to refer
to a Chamber of Deputies Public Hearing in the framework of the “discovery investigation
to evaluate the environmental impact of urban pollution, waste disposal and high risk
areas”. On that occasion, CNR delivered a complete report called “State of knowledge
CNR Environment and Health Inter-departmental Project
review on environment and health in high risk areas in Italy”.
The report includes the research activities carried out by the CNR Institutes in the 54
areas identified as Reclamation Sites of National Interest,(Italian acronym: RSNI) directly
managed by the Ministry of the Environment for the remediation activities. The research
activities were developed by 17 CNR Institutes, belonging to the following Departments:
Earth and Environment, Medicine, Materials and Devices, Molecular Design, Information
Communication Technology, Agrofood.
To give an idea of the size of this problem, it suffices to say that the population affected
by the impact of polluted areas is estimated in 6.4 million (but this number increases
up to 8.6 million if all the residents in the Municipalities of Milan and Turin, whose
territories are however only partly affected by RSNI, are included), living in the 54
RSNI that stretch themselves over the territory of 311 municipalities. These blunt figures
are important to assess the size (or – to help make a guess of the impact) of health adverse
effects that can be caused by substances recognised as hazardous and released in the
In fact, even modest risk increases, when acting on such large figures, can determine
very serious impacts in terms of the development of diseases, symptoms of indisposition,
mortality rates. Besides, since the residents of polluted areas are not equally exposed
and include socio-economically vulnerable people, or more susceptible people due to
a genetic predisposition or co-morbidities, the impact assessments are crucial. The size
of the exposed population and the risk intensity are the main criteria to assess the health
impact and, more in general, the impact on the public health service: in terms of possible
scenarios, the two extremes are represented by a situation in which very aggressive factors
act on limited groups of people (particularly exposed workers or small communities), or
by situations where risk factors have a weak action but act on a large number of exposed
The areas included in the RSNI sites are former industrial areas, caves, landfills; in those
cases the remediation procedures are long lasting and often very difficult to undertake from
a technical viewpoint; in the case of productive areas, remediation can be implemented
when an economic advantage exists, a change in production is required or strong pressure
from the public and administrators is exerted.
CNR activities covered a wide range of research sectors: monitoring, pollution
characterisation, remediation, epidemiology, planning.
The identification of pollution is the first step, carried out through different monitoring
devices in soil, sub-soil, fresh and sea water, indoor and outdoor air.
The pollution characterisation requires the use of devices such as a radioactive tracer
and mass spectrometry. In sea water environment, the characterisation is based upon
the assessment of the presence and conditions of the fish population, micro-organisms,
phytoplankton, sediments.
As concerns emissions into the atmosphere, the research includes studies on climate and
modelling, and studies on the relationship between indoor and outdoor pollution.
The removal of pollutants urges on the analyses of the remediation potential for water
and soil pollution, including bioremediation, phytoremediation and soil washing.
Emerging fields of research are represented by electromagnetic remote sensing,
electromagnetic diagnostics, passive remote sensing in optics, and methodologies for
automatic interpretation and integration in Geographic Information Systems (GIS),
Global Positioning System (GPS) and georeferencing systems.
Indicators to monitor environmental health effects need a wide range of data searching,
analysis and testing; epidemiological research can help to identify the existence of
environmental pressures, also using environmental data and GIS. Personal exposure can
be monitored using portable personal devices.
Health Impact Assessment procedures and experiences are finally presented in the Report
as a tool to be used for evaluation and public participation building in highly polluted
The Environment and Health Inter-departmental Project of CNR-PIAS
The CNR experience described above paved the way to build a multi-disciplinary
programme, to promote and coordinate collaborative research and joint actions on
environment and health.
Following the said report, the Earth and Environment and the Medicine Departments
launched a call for project ideas and proposals on environment and health research
activities to the whole network of CNR Institutes, taking as reference international
priorities and recent European developments.
130 proposals from 37 Institutes (from the Departments of Earth and Environment,
Medicine, Materials and Devices, Molecular Design, Information Communication
Technology, Agrofood, Energy and Transport) were submitted in few months, with a cost
estimation of approximately 30 million Euro.
In mid 2007, a project named Environment and Health Inter-departmental Project (Italian
acronym: PIAS-CNR) was presented to the CNR Board.
The actions and objectives planned in the PIAS have been selected taking into account the
EU and WHO Action Plans, the work presently carried out by European experts groups,
the indications found in national strategic documents and plans, the excellence of CNR
and the scientific and institutional collaborations that could be involved. The deliverables
to be accomplished in the framework of PIAS have also been planned accordingly to an
evolving real financial support.
The Environment and Health Project has the overall objective of promoting an integrated
research between these two scientific sectors, and of developing, in particular:
- knowledge of pollution sources and their consequent negative effects on health
- instruments and methods to analyse environment and health interactions
- instruments and methods to be used in managing complex situations.
On the base of the allocation of available funds to PIAS, the activities have been divided
into: start-up phase (phase 1) and research development phase (phase 2).
They will fulfil the following specific objectives:
1. To promote collaboration activities on environment and health among CNR
1a. to establish a database of the running projects promoted or implemented by CNR
Departments or in collaboration with other Institutions;
1b. to promote the creation of think thanks and project development groups;
CNR Environment and Health Inter-departmental Project
1c. to promote training workshops.
2. To facilitate the participation of CNR researchers in international experts’ Working
Groups on environment and health:
2a. to help disseminate information and to support the CNR participation in EU
Commission Working Tables on Environment and Health Strategy of the 7th
Research Framework Programme;
2b. to help disseminate information and to support the CNR participation in the WHO
work on Environment and Health, preparatory to the realisation of the Fifth Interministerial Conference on Environment and Health to be held in Italy in 2009.
3. To support the access to the funding schemes of the EU 7th Research Framework
Programme on Environment and Health:
3a. to help disseminate information and project drafting to get access to the EU 7th
Research Framework Programme on Environment and Health;
3b. to help disseminate information and project drafting to get access to other
international and national funding schemes, including Structural Funds 20072013.
4. To develop the present CNR competences on Environment and Health Research:
4a. To strengthen the research by selecting pilot projects on issues where collaborations
and competences of the various Institutes are more advanced and consolidated, and
whose results are considered as feasible with the available financial resources.
In the research development phase, the following scientific objectives must be achieved:
• to promote the transfer of research results to the production system;
• to promote the transfer of research results to the decision makers;
• to promote the experience of Health Impact Assessment, HIA, on present planning
instruments (including the Strategic Environmental Assessment, SEA) and the
production of methodological handbooks;
• to produce transferable results for the development of Environment and Health
supervision and monitoring systems;
• to test and validate environment and health indicators suggested by EU and the WHO,
supported by suitable information systems;
• to test and validate the use of georeferentiation tools (GIS) for a joint evaluation of
territorial health and environmental data;
• to test and validate instruments and methods of risk reporting in different frameworks
and to produce guidelines.
Project development
PIAS will be implemented through accurate and exhaustive examinations, development
of methods and field applications. It will take advantage of the network of scientific
partner institutes and a network of sites or areas in which studies and field research
have been carried out or are presently underway. The sample areas are selected as those
which show the most relevant health and environment impact issues (urban, rural, waste
disposal, reclamation areas, etc.).
This project is based upon multidisciplinary Working Packages (WP) and Organisation
Units (OU) in which CNR researchers, mainly from the Medicine and the Earth and
Environment Departments participate. Researchers from other Institutes with which CNR
collaborates (for instance, the WHO, the National Institute of Health (ISS), the
National Institute for Occupational Safety and Prevention (ISPELS), the Institute for
Environmental Protection and Research (ISPRA), the ENEA; Universities, the National
Health System, the Ministry of the Environment and Territory, the Ministry of Health, some
Regional Agencies for Environment Protection, ARPAs and Regional Epidemiological
Observatories are also involved in the PIAS.
The project is formed by two methodological modules for knowledge development (WP1,
WP2), an operative module representing the “engine” of the project (WP3), an interface
module to transfer technology innovation and industrial development results (WP4) and
an interface module to transfer communication, information and training results (WP5).
The OUs can operate in just one WP or in two or more WPs.
(Research areas, WP division, work orders, etc.)
Overall project structure and working packages (WP)
The relevance of the issue and the multidisciplinary research involved in the study of
mechanisms and methods as well as in technology application and development, make
this a project of primary interest not only for a national research council, but also for the
Ministries, such as the Research, the Health and the Environment ones, as well as for
other public and private Institutions.
Research Areas
1. Studies on environmental fate, biological perturbation mechanisms and health (WP1);
it is the basic module for the development of the necessary knowledge to produce
results to be tested in WP3.
1.1. Environmental quality assessment
• Lifecycle and the environmental fate of pollutants
• Chemical, physical and toxicological characterization of sub-systems
• Risks for ecosystems and human and animal health
• Detection of risk thresholds and effective health control levels
CNR Environment and Health Inter-departmental Project
• Exposure assessment and identification of exposed people
2. Analytical and methodological tools to meet the new environment and health challenges
(WP2); this is the basic module to develop the necessary methodologies and tools to
carry out the tests foreseen in WP3.
2.1. Monitoring and assessment tools
2.1.1 Environment-health indicators
• Bio-indicators of environmental quality, with reference to health risks;
• Bio-markers of exposure, physiological reaction, early damage of people’s
biological tissues, substances indicating health risk;
• Indicators of health consequences, diseases or conditions indicating adverse
effects associated to known or suspected environmental risk exposure
• Assessment indicators of actions/programmes/measures impacting the
primary interest effect classes.
2.1.2. Health impact assessment, cost-benefit analysis and other evaluation and
decision-making tools.
2.2. Measuring, supervision and recovery tools
2.2.1 Assessment and mitigation of health-impacting environmental effects;
2.2.2 Monitoring on environmental health indicators, ex-ante and ex-post
2.2.3 Instruments to measure, control and recover/improve the environment
and health. With particular reference to instruments measuring the exposure
dose (individual and environmental dosimetry) especially in the case of
combined exposure to different potentially hazardous agents.
3. Field surveys (WP3)
This is the core module, the “engine” of the project, oriented towards the field
experimentation of the outputs of the previous two working packages (WP1 and WP2),
focused on the development of knowledge and methodologies, and the preparation of
useful results for technological development and communication management as foreseen
in the subsequent packages (WP4 and WP5).
Another objective of the WP3 is the feed-back through knowledge development packages,
in particular the WP2, to produce materials, methods and tools for the development of
application research and to provide inputs to the WP4 and WP5.
The WP3 main objective is the assessment of the exposure, risks and effects on areas
and sites of primary interest for environmental health impact (urban, rural, industrial,
waste processing, reclamation areas, etc.). This package is structured in two different,
parallel and interconnected activity sectors dealing with knowledge implementation and
the experimentation of specific methods on the environment-health relation, starting
from the study of hazardous environmental factors or from the study of environmentally
sensitive diseases.
3.1 Environmental contamination that produces health effects
• inhaled fine particulate matter (PM10), (PM2.5) and ultrafine or nano particles
• heavy metals
• asbestos
• radon
dioxins and PCB (polychlorinated biphenyl)
endocrine disruptors
organic pollutants
electromagnetic fields, with particular reference to extremely low frequency
electromagnetic fields (ELF); mobile telephone systems and occupational
• relations between external and internal air pollutants
3.2 Environmentally sensitive diseases
• respiratory diseases, in vulnerable groups (children, senior citizens) and in
urban population
• bronchial asthma associated to the interaction of climatic and oxidizing pollutant
factors (NOX, O3)
• neurological development diseases
• adverse reproductive events and child and paediatric tumours
• effects of endocrine disruptors
• cardiovascular diseases as a consequence of air pollution exposure
• tumours and other diseases in adults as a consequence of environmental
exposure in uterus and early life
• genetic sensibility and gene-environment interaction
4. Technological and industrial development (WP4)
This is a cross-interface module for all project activities, used to provide technological
development for the outputs from previous modules.
4.1 Environmental monitoring
4.2 Reclamations
4.3 Biotechnologies
5. Information, communication and training (WP5)
This is a cross-interface module for project activities, used to communicate the outputs
from previous modules
5.1 Risk perception methods and tools
5.2 Risk communication methods and tools
5.3 Governance strengthening instruments (including regulatory suggestions)
5.4 Instruments to promote public accountability
5.5 Information and training methods and tools
PIAS implementation
PIAS implementation is closely linked to the funds available at each stage: the first phase,
now completed, allowed to explore the potentialities of a collaborative work, and to
conceive project developments, as described in the following chapters.
The implementation is based on a voluntary project agreement among the CNR Institutes
interested to work together on the topics identified as relevant. Working Groups and Pilot
Studies Groups have been identified after the call for project ideas and proposals.
Six Working Groups (WGs) were defined to study this complex chain, starting from the
environmental pollution to the emergence of illness. Several CNR Institutes are involved
in each WG, together with University and Public Bodies, where relevant.
WG1 – The fate of pollutants in soil (the chain of pollution from emission to human
CNR Environment and Health Inter-departmental Project
WG2 - Water and soil monitoring for the protection of environment and human health
(specific monitoring for the pollutants identified as dangerous for health)
WG3 - Role of atmospheric pollution on harmful health effects (the relation of outdoor
with indoor pollution, the molecular mechanisms of disease promotion)
WG4 - Human biomonitoring (biomarkers of exposure and early damage, the
relations among epidemiological studies, research in toxicology, in vivo and in vitro
WG5 – Environment and health surveillance systems (to develop a protocol for high risk
hot spots and selected areas)
WG6 – Monitoring contaminants in food chain and their impact on human health (the
pollution chain from the emission to the food chain, the pollution of animal feed)
Two Pilot Studies(PSs) are in progress. They focus on well-established issues, and are
in charge of drafting a feasibility project to apply for EU-FP7 or other call for proposals.
Several CNR Institutes are involved in each PS, together with the University and Public
Bodies, where relevant.
The Pilot Studies deals with the following challenging fields:
PS1 – Endocrine Disruptors and health effects
PS2 – Ultrafine particles and cardiopulmonary effects
WGs deliverables include: a workshop to put together experts and building capacities;
intermediate documents and working reports to share and discuss; a middle term national
workshop for WGs and PSGs together; a summary document including all the activities
and prospects for future research (this publication); a conclusive document including
the operational proposals and the pre-feasibility projects addressed to the CNR Board of
Directors; an international workshop, to be organised in late 2010.
One young researcher was hired as assistant for each WG, whereas PSG are operating to
advance research and define pre-feasibility projects.
The national PIAS workshop to compare and discuss the advancements after one year
of activity was held in June 2009 and it is now available on line, with video and audio
streaming at
National projects and international collaborations
The National Institute of Health (ISS) coordinates a Strategic Environment and Health
Programme, now in its second year of activity. The Programme is divided into six projects
for a total of 41 units and covers the health impact associated with living in polluted
sites, in areas affected by waste disposal/incineration facilities, and the exposure to air
pollution in urban areas. CNR researchers, together with other Institutions, participate in
the Programme implementation. The Programme included the following Projects: The role
of ultrafine particles in the pathogenic mechanisms of cardio-respiratory effects produced
by an urban pollution; Possible health effects of waste disposal in populations living
near disposal/incineration plants and comparative evaluation of the applied technologies;
Short-term effects of air pollution in urban areas: effects of gases and fine and ultrafine
particles, pollution-temperature interaction, individual susceptibility; long term effects of
air pollution: cohort studies in adults and children; Meteo-climatic conditions and health:
definition and identification of risk, effects measurement, the evaluation of intervention
effectiveness on epidemiologically relevant pathologies; Health risk in polluted sites:
exposure estimation, bio-monitoring, epidemiological characterisation.
The Institute for Environmental Protection and Research (ISPRA) is involved in several
activities concerning the environment and health, such as the dissemination of information
on environmental monitoring, novel risks and the promotion of Health in All Policies.
ISPRA is also part of the ERA-ENVHEALTH network.
The ERA-ENVHEALTH is one of the European Union network initiatives of the ERANET ‘family’. The project started in September 2008, and put together managers from 16
environment and health research programmes from 10 countries with the coordination of
AFSSET, the French Agency for Environmental and Occupational Health Safety. ERAENVHEALTH strategic objectives are: to establish a network of programme managers
and financers to share information and expertise on research; to define opportunities
for research cooperation and coordination; to identify priority areas for multinational
research; to develop coherent joint activities at the EU level; to implement joint multinational calls for specific research proposals on environment and health; to provide policy
support for the implementation of the Environment and Health Action Plan (2004-2010)
and other EU policies concerning the environment and health issue. The originality of
the ERA-ENVHEALTH resides in the promotion of a trans-national joint call in order to
experiment joint funding and to fully assess the implementation. The ERA-ENVHEALTH
fosters the use of environment and health research results to support policy development,
and supports the early identification of critical issues having a public impact.
The NRC and the Institute for Environmental Protection and Research are the Italian
partners of the Project, whereas the Environmental Protection Agency of the Tuscany
region (ARPAT) and the Regional Agency for Prevention and the Environment of the
Emilia-Romagna region (ARPA-ER) are Consultative organisations and collaborate to
spread information and to seek the participation of the scientific community.
The Environment and Health Inter-departmental Project of CNR (PIAS) promotes
interdisciplinary research on the interaction between environment and health.
PIAS established a network for collaboration and join activities among researchers coming
from different research institutes, that traditionally operate separately in environment
sciences or in health disciplines, and to bridge the gap towards an interdisciplinary
The project implementation confirmed the soundness of the original design. PIAS, with
its advances in knowledge and its results, represents now a valuable and durable platform
to start a broader research programme focused on field investigations and basic research
on environment and health, including studies on mechanisms, research on methods and
tools, risk communication and innovation for technology transfer.
The successful establishment of the PIAS Working Groups can be now considered as the
framework to evaluate several proposals and action plans to be implemented.
Its documented high scientific level, coordination ability and resources attraction,
competitive attitude, capacity building and networking have made it possible to complete
the priority-setting stage.
The consolidation and strengthening of the existing collaborations in the European and
international networks is one of the main goals of the PIAS Project to guarantee a top
level research and to offer advanced competences and instruments at the national level.
CNR Environment and Health Inter-departmental Project
The fate of pollutants in soil (WG1)
Since the soil has several functions directly related to human health, such as the production
of food as well as a filter action for groundwater, the preservation of its functionality
from any possible threat caused by human and natural events is clearly important. The
fate of contaminants in soil has to be addressed in order to evaluate the potential exposure
of people, taking into account the complexity of pathways, the interactions with soil
surfaces, the changes in the chemical and biological conditions of soil environment. The
study of soil environment can thus provide a basis for the assessment of human exposure
and health adverse effects. Research activities such as the dietary uptake of vegetables
grown in polluted soils, accidental soil ingestion, bio-accessibility and bioavailability
must be analysed. The case study of Gela (Sicily) industrial allowed a progress in our
knowledge of specific contamination sources (such as landfills or industrial sites) and
vulnerable groups, the latter studied on the basis of their place of residence, work activity
or dietary habits. Pollution pathways are strongly influenced by the chemical and physical
nature of soil, by its equilibrium in a thermodynamically open multiphase system. The
identification and understanding of the mechanisms linking soil quality and health is
proposed through an integrated approach.
Seven CNR Institutes plus one University Department are directly involved in the PIAS
Working Group.
Water and soil monitoring for the protection of the environment and human health
Current water and soil monitoring programmes are based on the sampling and laboratory
analysis of chemical and microbiological variables. Different methods to measure effects,
directly applied on living organisms, both at individual and at community level, have
been integrated into monitoring plans. Emerging problems are related to new classes of
pollutants, not yet regulated by legislation. The CNR Institutes carry out research on several
emerging environmental issues, such as engineered nanoparticles and perfluorinated
compounds in water environments, that have been chosen as case studies. An innovative
monitoring approach, ranging from the measurement of the effects to the identification
of causal molecular agents, is discussed. “Toxicity Identification and Evaluation” (TIE)
and “Effect Directed Analysis” (EDA) monitoring procedures were reviewed. In parallel
to “traditional” in-vitro tests, the development of the “omics” disciplines is fundamental
to study the relationships between the genome or protein structure and the activity and
biological effects of exogenous agents. Methods to identify the activity of dioxin-like
compounds as the cause of a specific adverse effect on river organisms are presented
and discussed. Applications and perspectives of investigative monitoring were examined
(in the case of an unknown agent or source for toxic or other biological effects) and of
screening (for risk assessment of specific pollution sources). All the above mentioned
activities are crucial for an effective management of the territory to safeguard human and
environmental health.
Seven CNR Institutes are directly involved in the PIAS Working Group.
The role of atmospheric pollution on harmful health effects (WG3)
Gaseous and particulate species in outdoor and indoor air play a key role in increasing
the morbidity or mortality observed in many clinical studies. The knowledge of the
main toxicity patterns of atmospheric pollutants needs to be improved, especially as
concerns particulate species. This is mainly due to the varying size-distribution, chemical
composition and different mechanisms of toxicity of fine and ultrafine particles (UFPs).
Recent findings on toxicity routes attributable to gases and particulate matter (PM)
species (i.e. the water-soluble organic fraction (WSOC) studied for the strong oxidative
potential to biological tissues) are reviewed. Toxicity routes are discussed to hypothesize
the relationships among sources, diffusion pathways, receptor sites and susceptible
populations. Strategic aspects are underlined, to be further developed in the ‘Feasibility
study for the assessment of the health effects of the chemical composition of ultrafine
particles’, presently in progress. The nature and role of aerosol particles and gaseous
mixtures are major research issues, due to their potential hazard for human health; the
connection of the toxicological and epidemiological impacts of atmospheric particulate
matter to its chemical composition is of paramount importance to assess effective pollution
abatement strategies. During a number of field experiments, state-of-the-art instruments
have been used for aerosol characterization.
Two CNR Institutes are directly involved in the PIAS Working Group.
Human Biomonitoring (WG4)
Human Biomonitoring (HBM) aims at identifying biomarkers useful to measure
environmental exposure, at monitoring its biological effects and the causal relationship
with pathological conditions, and at defining, where possible, the genetic susceptibility of
the overall population. The search of reliable biomarkers, i.e. objectively measured and
validated as health or disease indicators, requires the expertise of scientists with different
specializations, able to tackle increasingly complex problems through a multidisciplinary
approach. The PIAS work-package aims at promoting a scientific strategy to develop
and validate effect, exposure and susceptibility biomarkers. The chapter presents a
knowledge review, both in the basic and applied research, as well as future perspectives
and developments. Two main HBM objectives have been examined: i) the determination
of the levels of toxicants in biological fluids in the overall population; ii) the search
for new exposure, effect and susceptibility biomarkers. Four crucial points have been
tackled: the management of environment and health issues through a multidisciplinary
approach, the combination of medical tools with a biological approach based on
biochemistry, biophysics, cell and molecular biology, bioinformatics, molecular genetics
and genomics; the validation of conventional/new exposure biomarkers; the validation
of conventional/new effect biomarkers; the identification of genetic susceptibility
markers in the Italian population. The purposes of modern HBM have expanded beyond
their origin in occupational medicine to cover a wide variety of diagnostic procedures
and assessments of environmental pollution, leading to the identification of potentially
hazardous exposure before the evidence of adverse health effects. The definition of
exposure limits to minimize the likelihood of significant health outcome appears as the
final goal.
Four CNR Institutes plus one National Public Research Institute (ISPESL), one Scientific
CNR Environment and Health Inter-departmental Project
Foundation and one Hospital are directly involved in the PIAS Working Group.
Environment and health surveillance systems (WG5)
An integrated environmental and health surveillance system is the systematic, ongoing
collection and analysis of information related to disease and the environment (indicators)
and its dissemination to individuals and institutions. It is a scientific tool for the
implementation and evaluation of policies aimed at preventing, controlling and protecting
health and the environment. Different analytical approaches to classify environmental
and health indicators have been examined and discussed. A protocol to be tested in areas
with different environmental risks has been developed, in order to monitor environment
and health indicators and to provide useful tools for primary prevention programmes and
communication. The goal is to select a set of environmental and health indicators to be
assessed for their utility and availability in time and space.
Five CNR Institutes plus two Departments of the National Institute of Health, two Regional
Environment Protection Agencies and one Local Health Unit are directly involved in the
PIAS Working Group.
Monitoring the contaminants in the food chain and their impact on human health
A growing attention is paid in Europe to food safety and to the relation between diet and
consumer’s health. Changes in lifestyle, modification in food production and distribution
determine the eating habits of Western populations. Data from the annual report of the
European Commission Rapid Alert System for Food and Feed (RASFF), summarizing
notifications on food contaminations occurred in different countries, are useful to plan
efficient food control programmes. In this context, the PIAS working group studied how
specific classes of environmental contaminants (e.g. pesticides, metals, dioxins) may
affect human health through the food chain. Issues of major interest in this sector are, for
instance, the determination of heavy metals and dioxins in food matrixes and biological
samples; the existing experimental models to assess the harmful effects of contaminants
on human reproduction; the role played by the cytochrome P450 in the xenobiotics
metabolism. Finally, a research programme based on a holistic approach has been defined.
The proposal, whose target is the young population, aims at identifying the cause-effect
relationship between the presence of contaminants in the diet, their accumulation in
humans and the risk of developing chronic diseases. A discussion on the bioavailability
and adaptive response is presented, to suggest a possible functional link (at molecular
level) between the onset of specific diseases and the concentrations of contaminants
measured in food. An integrated approach to assess the impact of food contamination
on human health can increase our scientific knowledge and build consumers’ trust and
Three CNR Institutes plus one Public Research Agency are directly involved in the PIAS
Working Group.
The Pilot Study on ‘Endocrine Disruptors and health effects’ (PS1)
The pilot study focuses on the relationships among exposure to endocrine disruptors
and some selected diseases and the identification of how the environment and the diet
synergistically operate in promoting some severe pathologies in wildlife and humans.
Several experimental studies reported that also very low doses of endocrine disruptors can
affect the endocrine system, causing diseases and altering the development of mammalian
(humans included) and non-mammalian species. Cancer, cardiovascular risk, modulation
of adrenal, gonad and thyroid functions, and endometriosis are some of the diseases that
cause alarm in the citizens, associated to the exposure to endocrine disruptors. Research
activity focuses on three lines: i) the possible relationship among the levels of toxic
pollutants in biological fluids and the risk or occurrence of cardiovascular diseases, as
well as the alteration of thyroid, gonad and adrenal functions in the population of Gela
(an high environmental risk area); ii) in vivo experimental studies on endometriosis using
mice exposed to Bisphenol A, iii) measurements of endocrine disruptors concentration in
fish used for human consumption in the selected area.
Three CNR Institutes in collaboration with the Italian Endometriosis Foundation are
directly involved in the PIAS Working Group.
Pilot Study on ‘Ultrafine air particle and cardiopulmonary effects’ (PS2)
It aims at combining the results of two advanced activities in the identification of the
composition of atmospheric ultrafine particles and those of the health studies exploring
short-term effects of air pollutants exposure in subjects with selected diseases. Five workpackages focus on the improvement of knowledge in: i) the chemical composition of
ultrafine particles and their variability in urban and rural sites in Italy, based on available
multi-stage impact data and on initial measurements using Aerosol Mass Spectrometers,
ii) the methodologies to measure the oxidative potential of the water-soluble organic
fraction (WSOC) of the aerosol, iii) the short-term effects of exposure to air pollutants
in subjects with pre-existent arrhythmia, iv) the short-term effects of exposure to air
pollutants in subjects with pre-existent lung diseases, finally v) results are to be evaluated
to design an integrated Italian research activity for projects to be presented in the
framework of the available EU projects call for proposal.
Four CNR Institutes plus one University Department, one Regional Environment
Protection Agency and one Local Health Unit are directly involved in the PIAS Working
On the whole, eighty-six researchers belonging to nineteen CNR Institutes and other
twelve Public Research Bodies are directly involved in the PIAS project and have been
collaborating at the preparation of the present publication.
Fabrizio Bianchi
PIAS Coordinator, CNR - Institute of Clinical Physiology
[email protected]
Liliana Cori
PIAS Scientific Support, CNR - Institute of Clinical Physiology
[email protected]
Pier Francesco Moretti
CNR - Department of Earth and Environment
[email protected]
CNR Environment and Health Inter-departmental Project
The fate of pollutants in soil
G. Petruzzelli, F. Gorini, B. Pezzarossa, F. Pedron
CNR, Institute of the Ecosystem Studies (ISE), Pisa (Italy)
[email protected]
Different disciplines must be urgently put together to understand environmentally related diseases, and
to develop strategies capable of reducing the negative effects of pollution on human health. Within this
framework, the importance of soils and their characteristics on human health is receiving a growing interest
as the essentiality of soil for human life becomes increasingly clearer.
Understanding the transport and transformation of pollutants from the source of origin to the final receptors
(environmental ecosystems and humans), through different soil typologies, is of paramount importance.
On the basis of the duration, frequency and intensity of exposure, it is also important to evaluate which
concentrations are necessary to produce biological alterations in living organisms, leading to the onset of a
One of the main objectives of the Environment and Health Inter-departmental Project, PIAS-CNR, was to
highlight the close links between environmental matrices and human health. Within this framework, Working
Group 1 (WG1) focused attention on the fate of contaminants in the environment, particularly on the soil
ecosystem. The WG1 proposal aims to go beyond total diet studies and to understand mechanisms and
processes by which contaminants enter the food chain and influence to various extents nutrition and the
health of humans.
The integration of different disciplines can
help to overcome the compartmentalization
of increasingly specialized scientific
knowledge. This is urgently needed in
order to understand environmentally
related diseases, and to develop strategies
capable of reducing the negative effects
of pollution on human health. Within this
framework, the importance of soils and
their characteristics on human health has
been growing in interest as the essentiality
of soil for human life becomes increasingly
clearer (38, 46, 75).
Soil functions, such as food production,
are directly related to human health,. The
action of filters on groundwater highlights
the need to preserve soil given that its
efficiency can be reduced by human
activity and natural events (1, 108).
The fate of contaminants in soil is
important in terms of evaluating their
possible exposure to humans. Another
significant element is the complexity of
pathways determined by emission sources,
interactions with soil surfaces, and changes
over time in the chemical and biological
conditions in the environment where the
soil is located (Fig. 1).
Soil is defined as the top layer of the
earth’s crust and is made up of mineral
particles, organic matter, water, air and
living organisms. Soil is a multiphasic and
CNR Environment and Health Inter-departmental Project
Figure 1. Soil – health relationships
extremely dynamic system, with numerous
functions: it is the main producer of
biomass and raw material, it supports
biodiversity development (habitat, species,
etc.), it provides the main source of
carbon, and plays a fundamental role in
human activities and in the survival of the
The formation and regeneration of soil
are extremely slow and thus soil is
considered as a non renewable source.
(compaction, salinization, contamination,
impermeabilization, decrease in organic
matter, reduction in biodiversity), and
natural phenomena (erosion, flooding,
landslides). The resulting effects, however,
can be aggravated by human activities,
such as agricultural practices, industrial
activities, tourism, urban and industrial
development, and town and country
planning (1). These processes can lead to
a decrease in soil fertility, a loss of carbon
and biodiversity, a reduced ability to retain
water, an alteration in the gas and nutrient
cycles, and a less efficient degradation
of contaminants. Soil degradation has a
direct effect on water and air quality and
on climate changes. It can also influence
human health and present a danger in
terms of food safety (9, 33, 58).
Data analysis shows that soil degradation in
Europe may cost 38 billion euros per year.
The Thematic Strategy of the European
Union for soil protection (2006) proposes
guidelines aimed at protecting soil and
maintaining its ecological, economic, social
and cultural role (30, 31). The Strategy is
contained in the Plan of Environmental
Action of the European Community,
adopted July 2002 and valid until 2012. Its
priorities include climate change, nature
and biodiversity, the environment and
health, natural resources, and waste (32).
Co-ordinated action at a European level is
necessary in terms of the consequences of
soil degradation on other issues related to
the environment or food safety. Before this
directive was approved, soil had never been
the focus of European protection measures,
and soil protection was related only to
regulations concerning environmental
protection or other strategic fields, such as
agriculture and rural development.
The Strategy is a legislative bill which
permits the sustainable protection and
use of soil, integrates soil protection
within national and European politics,
and raises public awareness. According
to the directive, member states have to
take measures to avoid soil contamination
with dangerous substances and plan an
inventory of contaminated sites.
The fate of pollutants in soil
When chemical concentrations present a
risk for human health or the environment,
the directive calls on member states to
remediate the polluted lands, with the
aim of removing, controlling, edging or
reducing the pollutants. Similar strategies
are also followed at an international level
(25, 53, 74, 98, 99, 110, 132, 133, 134, 135,
139, 140, 141, 142, 143).
2.1 Soil and human health
Soil has always been vital to humans and
fundamental to human health since it is the
main resource for food production. The
link between the continuously increasing
world population and the ability of soil to
sustain that growth was the topic of Thomas
Maltus’s 1798 essay. The maintenance of
suitable nutritional food sources is an old
problem that is still present today. Soil is
not the only element that affects the food
supply, but it is an extremely important
resource needed in overcoming this
complex issue. In developing countries
characterized by a high rate of soil
degradation, the lack of safety regarding
an adequate dietary intake is a relevant
problem. Worldwide food production and
demand is likely to increase, making it
crucial to manage and conserve soil. One
of the the E.U objectives is to protect soils
against erosion and pollution since longterm productivity is likely to be affected
by soil degradation resulting in a relevant
reduction in yields on agricultural land,
not only in developing countries (77).
Soils may influence human health also
in other different ways. Ingestion of soil
may result in significant exposure to
toxic substances. Children are the object
of special interest, since soil adhered to
fingers may be inadvertently swallowed by
bringing the hands to the mouth, especially
during outdoor activities (68).
One of the most common effects of soil
ingestion is the alteration of the mineral
content and nutrient balance in individuals.
Ingested clays, due to the acidic
environment of the stomach, release the
elements contained within them through
the mechanism of cation exchange (82).
Concerning the toxicity of the contaminants
to which humans can be exposed through
the ingestion of soil, lead is a major
concern and focus of study. Children
are subject to a greater risk because lead
acts as a neurotoxin, with particularly
serious effects on the development of the
nervous system during childhood (96).
Soil ingestion plays an important role in
the risk assessment of contaminated sites,
where also soil inhalation is considered of
particular concern.
In the early 1980s studies revealed that
most of the soil dust inhaled by humans
is trapped and then swallowed, passing
through the gastro-intestinal tract.
However, a portion of this dust is trapped
inside the lungs, where it can progressively
lead to bronchitis, pneumoconiosis and
cancer of the lungs. The reaction of the
lungs to dust obviously depends on the
kind and the amounts of the dust inhaled
(137). Particles with a size comparable to
those of clay and originating from wind
erosion of the soil, once inhaled, can
settle in the pulmonary alveoli, causing
progressive inflammation in the lungs.
Further damages arise following inhalation
of particles coated with toxic substances
and also of the biotic components of soil
(20, 135). The fungus aspergillus present
in the soil is the biggest killer along with
the human AIDS virus, causing lung
infections following immunosuppression
(113). An infectious disease known as
“desert fever” is caused by inhaling spores
of the fungus Coccidioides immitis. In the
U.S, it is estimated that each year between
50,000 and 100,000 people are affected
CNR Environment and Health Inter-departmental Project
by symptoms of Coccidioidomycosis (7).
Tetanus is the most common desease to
potentially affect people who come into
contact with soil. This disease is due to
a toxin produced by Clostridium tetani
spores of anaerobic microorganisms. The
bacteria, present in the surface layer of soils
as well as in human and animal secretions,
are especially abundant in cultivated and
fertilized fields. Hookworm, characterized
by multiple clinical manifestations
including anemia, is a disease also
caused by skin contact with the soil and
whose hexogene agent is detectable in the
nematodes Ancylostoma duodenale and
Necator americanus. The survival of the
hookworm larvae in the soil is favored in
moist, sandy, crumbly environments and
at temperatures between 24 and 32 °C.
Infection can occur by oral ingestion of
contaminated food and, presumably, direct
ingestion of soil. The disease has a high
incidence in the rural areas of the tropics,
with higher occurrences in children (63).
Other diseases are ascribable to soil
characteristics. Podoconiosis has been
correlated with soils containing particles
of colloidal size derived from wethering of
basaltic rocks that are able to penetrate the
epidermis intact (119).
Soil quality largely determines ground
and surface water quality. In Bangladesh
drinking water is contaminated with
arsenic at concentrations of up to 1000 μg/l.
The consumption of contaminated water
led to the spread of disease and death, with
typical epidermal lesions (105). Considering
the sources of As and the mechanisms
that result in groundwater pollution, it is
possible that Fe hydroxides present in the
sediments are reduced by the activity of
microorganisms, favoring the release of As
absorbed in groundwater (92).
When the soil ability to retain organic
compounds by sorption processes is
reduced, groundwater pollution by organics
of industrial origin is a widespread problem.
The same happens for pesticides that can
penetrate into the soil through different
routes, such as root systems, leaves and the
decomposition of plant and animal tissue.
Other pesiticides are directly applied to
the soil and can be released from soil to
surface water and groundwater (120).
Many of these compounds are considered
endocrine disruptors and have severe
health implications.
Further contamination derived from
agriculture practices is the release of
the NO3- anion from soil. This ion has a
high solubility in aqueous environments,
and being negatively charged is poorly
absorbed by most soil surfaces. The
excessive presence of nutrients causes
algal bloom with serious consequences
on the whole aquatic ecosystem. Toxins
produced by rapidly growing cyanobacteria
can cause numerous human disorders,
pneumonia, allergic reactions, and liver
diseases including cancer (12). Among the
consequences of ingestion of nitrate ions
is childhood methemoglobinemia, which
severely damages the ability of hemoglobin
to carry oxygen in the blood (73).
The use of antibiotics applied to agricultural
crops for the control of plant diseases, and
their addition to animal feed have given
rise to several problems of immediate and
practical importance. The inactivation
of antibiotics in soil may be determined
by: intrinsic chemical instability of
the antibiotic molecule; adsorption on
soil clay minerals and organic matter;
antibiotics are a heterogeneous group
of compounds, varying greatly in their
chemical structure and reactivity, and soils
are not homogeneous, no generalizations
regarding the stability and biological
The fate of pollutants in soil
effects of antibiotics in soil are possible.
Although some antibiotics in soil are
unstable chemically, and many are
degraded microbiologically, it appears that
several antibiotics persist in some soils for
a time sufficient to produce harmful effect.
Soil bacteria are considered to be a source
of new resistance mechanisms to clinically
used antibiotics. In Europe, the livestock
industry consumes thousands of tons of
antibiotics per year. The application of cattle
manure to soil might be a relevant source
of antibiotics. The bacteria populations
resistant to antibiotics in soils are lower in
unmanured soils than in feedlot soils.
In the U.S, the presence in groundwater
of antibiotics such as tetracycline, added
to feed to promote livestock growth, may
present a possible means of determining
antibiotic resistance in humans. The
analysis of soil and ground water samples
from reserves close to farms have shown
that the bacteria are identical to those
in the gastrointestinal tract of animals,
and contain genes that are resistant to
antibiotics (27). This study suggests that
genes are transferred from bacteria of
the gastrointestinal tract of cattle to other
ecosystems. Since in the U.S. about 40%
of the water used for civilian consumption
comes from groundwater (and this value
has been gradually increasing), the
presence of antibiotics can lead to serious
consequences for human health (136). In
the EU, livestock consumes approximately
5000 tones of antibiotics each year.
Though there is no set limit on the use
of medicines in agriculture, veterinary
authorities have ruled that any compound
that can be accumulated at concentrations
higher than 7.5 g per hectare must undergo
environmental impact studies (114).
The deposition of feces in the soil from
humans and animals may potentially
contaminate fresh water sources with
bacteria, protozoa and viruses (122).
For example, Escherichia coli 0157 is a
virulent pathogen that in humans gives
rise to a broad spectrum of symptoms,
including hemorrhagic colitis (83). Cattle
are the main reservoir of the bacterium,
which, once reaching the ground, remains
there for several months. The most
common causes of infection from E. coli
0157 are associated with the consumption
of contaminated meat and dairy products,
although infection in humans may also
occur due to the contamination of soil and
drinking water.
A noteworthy amount of metals has
been released into the environment by
anthropogenic activities, in particular by
industrial processes and persist in the soil
due to their non biodegradability. Heavy
metal pollution is responsible for many
negative consequences both for human
health and the environment (17, 59, 61, 72).
Most heavy metals are considered essential
micronutrients and each of them requires
an adequate daily intake. However trace
elements are toxic if there are excessive
amounts of them in the human body, and
they have adverse physiological effects
at relatively low concentrations. Soil
ingestion represents a direct route for
the elements to humans. The transfer
of many elements from soil through
the food chain is an important although
indirect mean of exposure. Consequently,
deficiencies, excesses or imbalances of
inorganic elements from food sources
may have important consequences. An
inadequate intake of microelements is
recognized as an important contributor
to the global burden of disease through
increased rates of illness and death from
infectious diseases, and of disability such
as mental impairment (16). An increase
in the concentrations of microelements in
soil derived from weathering processes
CNR Environment and Health Inter-departmental Project
of the parent rock material or by human
activities such as industrialization, mining,
agricultural practices, and urbanization,
can cause an excessive release of elements
in the food chain and can have implications
on human health. The itai-itai syndrome is
probably the best known example of metal
contamination in the soil that has some
implications for human health through
the ingestion of contaminated food. It
developed in Japan in the 1950s, and it
is caused by food, especially rice, and
drinking water contaminated by Cd (35,
97, 111).
As a consequence, all legislations
concerning soil strictly regulate the soil
cadmium content to avoid its accumulation
in agricultural crops. However, Cd
accumulation in plants is determined by
the available fractions of metals in soil
rather than their total content. Although
the soils may contain high concentrations
of metals or organic contaminants, factors
such as pH, clay content, and organic
matter impact on speciation, mobility and
bioavailability of pollutants, influencing.
the amount absorbed by animals and
humans (131).
2.2 The fate of contaminants in soil
Soil contamination occurs through
either point source or diffuse pollution;
the main difference between the two
types of contamination lies in how the
contaminants are transferred to the soil.
Point sources, such as manufacturers,
landfills, incinerators, use soil as a
support and are linked to the activities that
necessarily transfer pollutants into the soil
(64). Diffuse sources are associated with
natural phenomena (long range transport,
atmospheric deposition, sedimentation
by surface water), with agricultural
practices, with recycling and inadequate
waste treatments. The most dangerous
contaminants in soil are, in general,
persistent organic pollutants (POPs) and
inorganic pollutants, above all heavy
metals. Persistent organic pollutants have
an anthropic origin and are characterized
by high lipoaffinity, semivolatility and
resistance to degradation. In the case of
heavy metals, that cannot be degraded or
destroyed, the presence in the soil could
be due to natural processes, for example
the formation of soil, and to anthropogenic
activities. Some are important essential
elements (Cu, Fe, Mn, Zn, Co), if present in
optimal concentration ranges, while others
(Hg, Pb, Cd) are potentially toxic elements
(19, 78, 80, 81, 84, 109, 130).
2.2.1 The nature and behavior of inorganic
Heavy metals are one of the numerous
classes of substances that can reach critical
levels in terms of human health, food safety,
soil fertility and ecological risks (80, 126).
Heavy metals are common contaminants
in the soil and bioaccumulate, thus their
concentration in the organism increases
over time compared to the level measured
in the environment. This is because the
absorption rate is higher than the excretion
rate in the organism (128).
The distribution of heavy metals between
the solid phase and the soil solution is
considered to be the key factor when
assessing the environmental consequences
of the accumulation of metals in the soil
(2, 69).
A physical and chemical analysis along with
an analysis of the soil profile is essential
for assessing the soil as a barrier against
inorganic contaminants, particularly heavy
metals (121). The retention of heavy metals
in the solid phase of the soil is dependent
primarily on the pH, and is linked to clay
minerals, humic substances, iron oxides
and hydroxides, and manganese found in
The fate of pollutants in soil
the soil, which all control the attenuation
effect even on anionic forms (116).
The retention and release process of
heavy metals includes precipitation and
decomposition, ionic exchange, and
adsorption and desorption.
The precipitation/release reactions may
involve discrete solid phases or solid
phases, which are absorbed onto the soil
surface. The ion-exchange reactions derive
from an exchange between an ionic species
in the soil solution and an ionic species
retained in sites with permanent charge
on the soil surface. The absorption and
desorption processes can affect all ionic or
molecular species and generally concern
absorbent sites with a pH-dependent
charge. These surfaces are iron, aluminum
and manganese oxides and hydroxides,
clay minerals and humic substances.
pH is the most important parameter
governing concentrations of metals in
soil solutions that regulate precipitation–
dissolution phenomena. Metal solubility
tends to decrease at a higher pH. In
alkaline conditions the precipitation of
solid phases diminishes the concentration
of metal ions in solutions and the reverse
happens with a lower pH. pH values
also regulate specific adsorption and
complexation processes. The sorption of
metals is often directly proportional to soil
pH due to the competition of H+ (and Al3+)
ions for adsorption sites, however this
competition may be reduced by specific
adsorption. Metal hydrolysis at higher pH
values also promotes the adsorption of the
resulting metal hydroxo complexes, which
beyond a threshold pH level (which is
specific for each metal) drastically reduce
the concentration of metal ions in the soil
solution. At low pH levels, on the other
hand, sorption processes are reduced due
to the acid catalysed dissolution of oxides
and their sorption sites, whereas the
complexation by organic matter tends to
decrease with increasing acidity.
Clay content
Ion exchange and specific adsorption are
the mechanisms by which clay minerals
adsorb metal ions. This is done through
the adsorption of hydroxyl ions followed
by the attachment of the metal ion to the
clay by linking to the adsorbed hydroxyl
ions or directly to sites created by proton
removal. Highly selective sorption occurs
at the mineral edges. However notable
differences exist among clay minerals in
their ability to retain heavy metals which
are more strongly adsorbed by kaolinite
than montmorillonite. This is probably due
to a higher amount of weakly acidic edge
sites on kaolinite surfaces. In expandable
clays (vermiculite and smectite) the
sorption processes essentially involve the
inter-layer spaces, and are greater than in
non-expandable clays such as kaolinite.
The importance of clay minerals, and of
soil texture in determining the distribution
of heavy metals between the solid and
the liquid phases of soil has direct
consequences on the metal bioavailability
of plants. For the same total concentration
it is well known that heavy metals are
more soluble and plant available in sandy
soil than in clay soil.
Organic matter content
The organic matter content of soils is
often small compared to clay. However,
the organic fraction has a great influence
on metal mobility and bioavailability due
to the tendency of metals to bind with
humic compounds in both the solid and
solution phases in soil. The formation of
soluble complexes with organic matter, in
particular the fulvic fraction, is responsible
CNR Environment and Health Inter-departmental Project
for increasing the metal content of soil
solutions. However higher molecular weight
humic acids can greatly reduce heavy
metal bioavailability due to the strength
of the linkages. Both complexation and
adsorption mechanisms are involved in the
linking of metals by organic matter thus
including inner sphere reactions and ion
exchange. Negatively-charged functional
groups (phenol, carboxyl, amino groups
etc.) are essential in metals retained by
organic matter. The increase in these
functional groups during humification
produces an increase in the stability of
metal organic complexes, which also show
a greater stability at higher pH values.
Cation exchange capacity
The density of negative charges on the
surfaces of soil colloids defines the CEC
of soil. This capacity is governed by the
type of clay and amount of organic colloids
present in the soil. Montmorillonitic
type clays have a higher net electrical
charge than kaolinitic type clays;
consequently, they have a higher cation
exchange capacity. Soils containing a high
percentage of organic matter also tend
to have high cation exchange capacities.
The surface negative charges may be pH
dependent or permanent, and to maintain
electroneutrality they are reversibly
balanced by equal amounts of cations
from the soil solution. Weak electrostatic
bonds link cations to soil surfaces, and
heavy metals can easily substitute alkaline
cations on these surfaces by exchange
reactions. Moreover, specific adsorption
promotes the retention of heavy metals,
also by partially covalent bonds, although
major alkaline cations are present in soil
solutions at much greater concentrations.
Redox potential
Reduction-oxidation reactions in soils are
controlled by the aqueous free electron
activity pE often expressed as Eh redox
potential. High levels of Eh are encountered
in dry, well aerated soils, while soils with
a high content of organic matter or subject
to waterlogging tend to have low Eh
values. Low Eh values generally promote
the solubility of heavy metals. This can
be ascribed to the dissolution of Fe–Mn
oxyhydroxides under reducing conditions
resulting in the release of adsorbed metals.
However under anaerobic conditions, the
solubility of metals could decrease when
sulphides are formed from sulphates.
Differences in individual metal behaviour
and soil characteristics result in conflicting
reports regarding the effects of redox
conditions on metal solubility.
Iron and manganese oxides
Hydrous Fe and Mn oxides, are particularly
effective in influencing metal solubility in
relatively oxidising conditions. They are
important in reducing metal concentrations
in soil solution by both specific adsorption
reactions and precipitation. Although
Mn oxides are typically less abundant in
soils than Fe oxides, they are particularly
involved in sorption reactions with heavy
metals. Mn oxides also adsorb heavy
metals more strongly, thus reducing
their mobility. This action is particularly
important in contaminated soils. Specific
adsorption of metals by hydrous oxides
follows the preferential order: Pb > Cu >>
Zn > Cd.
Other factors
There are a number of other factors
which may affect the solubility of metals
in soils. Temperature, which influences
the decomposition of organic matter, can
modify the mobilisation of organo-metal
complexes and consequently plant uptake.
An increase in the ionic strength of soil
The fate of pollutants in soil
solutions reduces the sorption of heavy
metals by soil surfaces due to the increased
competition from alkaline metals. Similar
effects also derive from the simultaneous
presence in soil solutions of many heavy
metals which compete for the same
sorption sites. This results in an increase in
mobility in contaminated soils due to the
saturation of adsorption sites. The living
phase of soil is also of great importance
in determining metal solubility, which is
dependent to some extent both on microbial
and root activity. In the rhizosphere, plants
can increase metal mobility by increasing
their solubility. This happens following
the release in the exudates both of protons
which increase the acidity, and organic
substances which act as complexing
agents. Microbial biomass may promote
the removal of heavy metals from soil
solutions by precipitation as sulphides and
by sorption processes on new available
surfaces characterized by organic
functional groups.
2.2.2 Organic pollutants
Among the many organic compounds
present in soil, the most dangerous are the
“persistent organic pollutants” that derive,
in general, from anthropic activity, are extremely persistent in the environment and
are transported for long distances (5, 28,
55, 66, 71). In specific environmental conditions they bioaccumulate and biomagnify, reaching considerable concentrations
that represent a threat for human health
and ecosystems. Of the twelve groups of
persistent organic pollutants, the following
three are acknowledged internationally:
polychlorinated biphenyls (PCBs), polychlorinated dibenzodioxins (PCDDs) and
polychlorinated dibenzofurans (PCDFs).
PCBs are high hydrophobic extremely
stable compounds and have very good
dielectric and thermostability properties;
these characteristics led to the diffusion
of PCB for industrial and civil use (8).
After accidental ingestion or due to their
presence in food compounds, PCBs are
absorbed through the gastrointestinal
tract, and then accumulate in body fats as a
consequence of their hydrophobicity (85).
The International Agency for Research
on Cancer (IARC) has classified PCBs as
potential carcinogenic agents for humans:
experimental tests suggest, in fact, that
these compounds may increase the risk of
skin, liver and brain cancer (24). In order to
protect human health and the preservation
of the environment, the European
Community banned the commercial use of
PCBs in 1990. However, these persistent
compounds are still present both in natural
soils, owing to long-distance transport,
and in soils that have been contaminated
by specific industrial activities (13).
PCDDs and PCDFs, which are generally
known as “dioxins” (118), are the undesired
by-products of chemical and combustion
processes and are also produced from
natural events, such as accidental fires and
volcanic eruptions. The dioxins are a group
of 210 chlorine-containing chemicals,
17 of which have a toxicological interest
owing to their carcinogenic potential and
their effects on reproductive, endocrine
and immune systems (48). Owing to their
high persistence in the environment, they
remain in soil, which become pollutant
reservoirs (117). In humans, the main
route of exposure to dioxins is through
food, which represents 90% of the total
exposure (51, 87).
EDCs. Over the last few years there has
been an increasing interest in identifying
the long-term damage to reproduction and
development; xenobiotics with potential
endocrine activities or endocrine disrupter
chemicals (EDCs) have been identified
as the main possible risk factors (47).
CNR Environment and Health Inter-departmental Project
Endocrine disrupters are a heterogeneous
group of persistent organic and inorganic
pollutants including dioxins, PCBs,
pesticides, and industrial compounds.
They are characterized by their potential to
affect the correct functions of the endocrine
system, especially the homeostasis of sexual
and thyroid hormones (10, 29, 125). These
molecules may enter the soil environment
by agricultural practices or industrial
waste disposal. The risks derived from
EDCs are determined by the distribution
of these compounds among the soil phases.
Depending on the chemical properties of
the molecules, EDCs can be either strongly
retained by solid soil phases, or leached to
deeper layers. Their mobility is largely
determined by adsorption – desorption
processes on solid soil phases.
A probable role of endocrine disrupters is
attributable to polybrominated byphenyls
(PBDEs), a class of manufactured
chemicals structurally similar to PCBs,
which were used in the past as flame
retardants (15, 40). Even though most
PBDEs were banned within the European
Union in 2006, studies have revealed that
PBDE levels have increased both in the
environment and in human tissues and
body fluids (34, 37, 50, 60).
Pesticides are a class of compounds used
to kill harmful organisms, especially in
agriculture. However many are also toxic
for other organisms, including humans
(93). The presence and bioavailability of
pesticides in soil can adversely impact soil
quality with related consequences on water
and air quality. Soil characteristics regulate
the processes that affect the behavior of
pesticides such as adsorption, degradation,
volatilization adsorption by crops. Pesticide
adsorption to soil depends on both the
chemical properties of the pesticide and
properties of the soil, in particular organic
matter. Organochlorinated pesticides have
been used for many decades and one of
their main features is their high persistence
in soil and transfer into the food chain,
with the consequence of well known toxic
effects in biota (67).
Behavior of organic contaminants
Organic molecules in soil are a carbon
source for microorganisms. Therefore, the
conditions that influence the breakdown
of organics by microflora should be
considered. Microflora are not always able
to attack organic molecules and digest them
completely, but often only partially break
them down. This results in compounds that
are even more toxic than the initial ones.
The intrinsic toxicity and health risks
following the ingestion of organic
compounds are well known, both natural
compounds and those deriving from
productive processes. On the other
hand, there is less information about the
potential contamination, caused by organic
compounds present in the soil, on the food
chain (plants-animals-humans). Organic
compounds should be evaluated in terms
of their chemical properties and their
relative absorption potential by plants, but
also in terms of the influence that the soil
has on them. In fact, these compounds
can be volatized, absorbed and therefore
immobilized, or transported along the soil
profile even to underground water.
The most important chemical properties
of organic molecules are those dealing
with their absorption in the food chain:
the distribution coefficient (octanol/water
(Kow), the Henry constant, solubility,
half-life, and the bioconcentration factor
The behavior of an organic contaminant
in the soil depends on the interactions that
are established with the solid, liquid and
gas phases of the soil, and with the living
phase. These relations give rise to the major
The fate of pollutants in soil
phenomena that rule the fate of the organic
contaminants concerning adsorption,
biotic and abiotic decay, leaching and
Adsorption and desorption
The adsorption processes of organic
compounds on the active surfaces of the
soil are particularly important because
they delay mobilization and leaching of
organic contaminants.
The distribution of the contaminants
between the liquid and solid phase of the
soil can be synthetically described by
the distribution coefficient Kd, which in
turn can be expressed as a function of
organic carbon (Koc) and of Kow. The
compounds that have high levels of Kow
and low solubility will be mostly retained
by the soil surfaces and be less available to
environmental processes.
Biodegradation is the most important
mechanism for the removal of organic
compounds in the soil. Degradation by the
microbial flora can increase the solubility
and therefore the availability of recalcitrant
compounds for microorganisms in the
soil. The chemical characteristics of
each specific compound affect the time
required for biodegradation. Various
parameters have been identified that could
be correlated with the degradation period.
For example, the half-life of polycyclic
aromatic hydrocarbons, PCBs and dioxins
are all related to the Kow. This coefficient
is also related to the leaching process and to
the persistence of contaminants in the soil.
In fact, compounds characterized by a log
Kow > 4.0 rarely mobilize. Therefore the
same compounds mentioned above, as well
as several organochlorinated pesticides
are very persistent and have a very low
leaching potential. Monocyclic aromatic
hydrocarbons, some chlorobenzenes, short
chain aliphatic compounds and phenols,
on the other hand, degrade rapidly and are
more easily leached from the soil.
Photolysis, hydrolysis and oxidation
(abiotic degradation) also contribute to the
disappearance of some organic compounds.
These reactions mostly affect compounds
with simple molecular structures, such
as phenols and some polycyclic aromatic
hydrocarbons (PAHs) with less than four
benzene rings. Volatilization also affects
volatile substances, which are generally
characterized by a reduced molecular
2.2.3 Soil - animal transfer
Depending on the grazing practice, the
season and the diet, both organic and
inorganic compounds present in soil
with a high level of contamination are
potentially transferable to animals. In
fact, contaminated soil is usually ingested
directly during grazing. The average
amount of soil ingested by most cattle
is around 6% of total removal (d.w, dry
weight), i.e. for a typical daily bovine
consumption of 15 kg d.w. it can reach 0.9
kg of contaminated soil per day. Assuming
that organic compounds in polluted soils
are present in concentrations from 0.1 to
10 mg/kg, the amount ingested can vary
from 30 to 3000 mg/year
Bioconcentration processes are particularly
important for persistent and non-polar
compounds (low solubility and with high
Kow). The BFC bioconcentration factors
related to diet, however, are difficult to
quantify experimentally. Nonetheless,
some models based on the daily intake
of organic compounds can provide an
estimate of their possible presence in meat
or milk.
When ingested these compounds can pass
through the gastrointestinal membrane,
CNR Environment and Health Inter-departmental Project
enter the blood or lymphatic system or into
some organs in relation to the lipid content
and, depending on the compound (PCBs,
dioxins, hexachlorobenzene), may have
long half-lives.
Other compounds such as polycyclic
aromatic hydrocarbons are not particularly
absorbed, but can be partially degraded
with the consequent formation of very
dangerous intermediate products.
Among foodstuffs, milk is especially
sensitive to organic compounds which
undergo considerable changes in the
concentration of organic compounds even
in response to short term changes.
2.2.4. Concluding remarks
More research is needed in order to evaluate
if the pollutant levels in the environment
threaten human health, considering the
most susceptible situations (proximity to
contamination sources such as landfills or
contaminated sites) and the most exposed
people, e.g. owing to work activities or
diet (6, 23, 54, 57). One very important
aspect is an understanding of the transport
and transformation of pollutants from
the source of origin to the final receptors
(environmental ecosystems and humans),
through the different soil typologies
(64, 103, 106, 129)). On the basis of the
duration, frequency and intensity of
exposure, it is also important to evaluate
the concentrations necessary to produce
biological alterations in living organisms,
until the onset of a pathology.
Diet represents the principal means
through which not only chemicals, but
also micro-organisms and mycotoxins,
reach humans (52). Research is needed on
the bioavailability mechanisms of organic
and inorganic pollutants in soil, and the
survival and persistence quantification
of pathogen agents in the environment.
The detection of microbic interactions
is also essential, together with further
knowledge of emerging pathologies and
molecular toxicology. Finally, studies need
to be performed on the long-term effects
following chronic exposure to low levels
of individual or mixed chemicals, as well
as the consequences of exposure to high
concentrations of natural elements.
3.1 Soil characteristics of the Gela site
Gela is a town in Sicily located in an area
of important industrial activity, which has
caused over the course of time significant
contamination of the environment. In fact,
it is so polluted that the Italian government
has designated it as an area (in Italian
known as “Site of National Interest: SIN”)
that is subject to specific regulations in
terms of remediation. The Gela site lends
itself to a new interpretation of pollution
that is no longer confined to a contaminated
site, but which has instead spread to a
wider environmental area. The Institute
of the Ecosystem Studies (CNR-ISE, Pisa)
have tried to highlight how important
soil characteristics are in defining the
contamination pathways of pollutants, and
how complicated it is to establish general
relations since each type of soil has specific
characteristics that differentiates it from
other types.
Given the complexity of the matrix,
which can contain many organic and
inorganic contaminants, the definition
of a potential hazard based on soil
characteristics, is limited. It is only
thorough a characterization of the soil
of the industrial area of Gela affected by
pollution that it would it be possible to
give a more accurate response. However,
The fate of pollutants in soil
given the amount of data produced during
the characterization phase of the site, we
planned to assess the characteristics of the
soils derived from analysis certificates,
based on the analytes determined. We also
wanted to evaluate which soil parameters
were missing, which are useful to detect
contamination pathways.
As mentioned previously, the potential
hazards of a contaminant present in the soil
and its risks to human health, particularly
through the exposure pathways from the
soil through the food chain and finally to
man, can be better defined if we know the
chemical and physical properties of the
soils in which contaminants are present (1,
2, 22).
We examined the documentation collected
at the Italian Ministry of the Environment
on Gela site to check whether, in addition
to the characteristic parameters of
contamination (pollutants concentration),
there were also parameters describing
the characteristics of the soils. This was
because these factors would help to predict
the environmental mobility and potential
bioavailability of the contaminants
The parameters that were observed and
that are most frequently reported are
pH, cation exchange capacity (CEC) and
organic carbon (C). These quantities are
fundamental to understand what types of
soil are in the areas concerned and how
they interact with the contaminants in
the soil. These data are shown in Figures
2, 3 and 4. However, there are no data on
texture, which is of paramount importance.
Such data could possibly be obtained,
even if only partially, from the geological
description of any probing that might have
been done.
Given the significant amounts of data on
the characterization of soils within the
various industrial areas, some conclusions
can be drawn with regard to heavy metals.
However, based on the data reported on
the certificates, it is not possible to make
concrete hypotheses regarding organic
contaminants. Among the parameters
identified, pH is particularly constant and
depends on the nature of the mineralogical
substrate from which the soil originates.
In fact, it is the most important parameter
that governs the concentration of inorganic
elements in soil solutions. In the soils from
the Gela area there should both be a limited
mobility of heavy metals, which move like
ions with positive charges (Cd, Zn, Cu,
Pb), and a limited bioavailability of such
metals. The same should hold true for
mercury, however the probable presence
of high concentrations of chloride ions
can greatly facilitate the mobilization
of the element and its diffusion in the
environment. Metals that move like ions on
the other hand, with a negative charge such
as As, may be more easily mobilized, and
could become a significant problem in the
whole area. The other parameter reported
in the characterization analysis of soil is
the cation exchange capacity. This quantity
expresses the charge density on the surfaces
of soil colloids. It varied considerably
from one industry to another which meant
it was impossible to identify a uniform
retention capacity of metals in different
parts of the Gela area under investigation.
The importance of pH and CSC is not as
significant in organic compounds as it is in
inorganic contaminants. These parameters
have a limited influence on the mobility
of nonionic organic compounds, which
are much more influenced by the content
of organic matter. Organic matter in the
soils of this area appears to be quite low
although there are considerable differences
between one area and another. Variability
in the values of organic matter and the
lack of soil texture characteristics are of
CNR Environment and Health Inter-departmental Project
primary importance for understanding the
behavior of organic contaminants. This
highlights the need to integrate data from
the characterization of the site with the
features of the soils, which could be partly
drawn from land use maps at a provincial
or regional scale. The characterization
of environmental matrixes can be a key
issue for population exposure estimates,
integrated with data on health and
epidemiology. As far as soil is concerned,
in addition to the commonly used ways
of evaluating a contaminated site, such
as skin contact and direct ingestion, it is
also necessary to take into account the
characteristics of this environmental
matrix affected by the contamination. Soil
properties determine the movement of
pollutants and their passage into the food
chain (1). This leads to an understanding
of low-dosage effects, which are prolonged
over time and which are often forgotten
in decontamination strategies dictated by
the need to solve immediate and acute
problems resulting from pollution.
By understanding the specific soil
characteristics of the area, if possible along
with a related food analysis, uncertainty
in the exposure calculations may be
reduced and a relationship can be defined
between the sources and targets of the
In order to evaluate the capacity of the
soil to interact with different types of
pollutants, it is necessary to consider
this environmental matrix as a threephase system. In general, the solid phase
constitutes 50% of the soil, while the other
half is made up of a porous space which, in
a good quality soil, contains half water and
half air. The solid phase, the degradation of
the parent rock, contains organic materials
(humic substances) that are concentrated in
the upper layers. It also contains inorganic
materials which at a certain depth become
the exclusive constituents of this phase.
The liquid phase is made up of water
that forms a “soil solution”. This solution
contains dissolved substances and can
dissolve other substances from the solid
phase. The soil solution reaches the roots
and gets into pores. This is the principle
means of transport of all of the substances,
including the pollutants. The gaseous
phase of the soil is made up of air, which
on the surface layer is richer in carbon
dioxide because of the high quantities of
organic material.
The flow of air into and out of the soil
is essential for plant growth and for the
decomposition processes of animal and
plant residues, as well as all materials of
an organic nature.
Mercury is particularly important in the
Gela site. Mercury, like other metals in the
soil, may be present in a dissolved form
as a free ion, or absorbed non-specifically
by weak electrostatic bonds, specifically
absorbed by covalent bonds, made more
complex by organic matter, or precipitated
in its solid phase in the form of carbonate,
hydroxide or sulfide (18). Depending
also on redox conditions, mercury can
exist in three valence states, Hg0, Hg and
Hg2+. Its bivalent form is generally highly
reactive with dissolved ligands, and is
highly soluble in water. It very often forms
complexes with Cl-, OH-, S2- and with
sulfur-containing functional groups of
organic compounds and NH3.
Mercury also forms complexes of
moderate stability with Br- and I- and some
nitrogenous R-NH2-type binding agents.
The factors that control the speciation
of the metal in solutions are pH, ionic
strength, redox potential, a concentration
of dissolved organic matter (DOM),
and dissolved ions such as oxygen and
The maximum solubility of mercury occurs
The fate of pollutants in soil
in an oxygenated environment (Eh 350-400
mV) which is the typical condition found
in soil. The principle forms that are found
in soil are Hg(OH)2 and HgCl2. With these
ions, mercury can form soluble complexes
that are environmentally significant
because they are very mobile. On the other
hand, in anoxic environments these ions
form stable and insoluble sulfides.
Methylmercury, CH3Hg+, and dimethylmercury, (CH3)2Hg, are also formed in the soil
(49, 90) but they constitute on average less than
2% of the mercury present in the soil. Even at
low concentrations, these compounds can cause
serious bioaccumulation problems (101).
CH3Hg+ is synthesized by microbe activity
(bacteria and fungi) both aerobically
and anaerobically. It is soluble in water
and forms different compounds such as
CH3HgCl, CH3HgOH and CH3HgSH.
The anion that binds itself is particularly
important because it determines the
biological uptake. The speciation of the
CH3Hg+ ion is similar to Hg2+ and therefore
the parameters that influence it are the
same: pH, DOM and ionic strength.
Mercury is mobilized in the soil through the
formation of soluble inorganic compounds
which include HgCl2 and Hg(OH)2. The
degree of mobility of these complexes
depends on the type of charge and on the
chemical and physical characteristics of the
soils in the area. The presence of chloride
ions makes the metal highly mobile for the
formation of very soluble complexes.
In the presence of high amounts of organic
substances, a process that is equally
important is the formation of organic
complexes of bivalent mercury due to the
high affinity of the Hg(II) ion and of its
inorganic compounds for the functional
groups containing sulfur. A part of bivalent
mercury can be complexed by soluble
humic substances, such as fulvic acids, and
therefore may be present in the liquid phase
of the soil. Mercury loss due to ground
runoff is still very small compared to the
total percentage, so that in contaminated
soils like those in the Gela area the metal
can be expected to be released for a very
long period, thus affecting human health
for many years.
While historically mercury has been the
element of greatest concern in the area
of Gela, the soil characteristics are more
favorable to provide conditions for a great
mobility and bioavailability of arsenic. Like
other metals arsenic toxicity depends on
the chemical form of the element, organic
compounds being much less toxic than
inorganic ones. The main forms of arsenic
in soil are arsenate and arsenite which
are highly toxic, because their molecular
similitude to phosphate can interfere with
the functions of many proteins. USEPA
defined arsenic as a human carcinogenic
Soil arsenic may influence human health
by soil dust respiration, soil ingestion and
consumption of contaminated water (3).
Moreover arsenic may enter the food chain
via crops and vegetables grown in polluted
soil (62).
The total content of arsenic in soil is
not a reliable indicator of the potential
hazards for health and environment. Its
mobility and bioavailability are largely
determined by soil characteristics (104).
The retention of arsenic in the solid
phase depends on soil pH, mineralogical
composition and competing ions in soil
solution. In aerobic soils, sorption on
metal oxides is the main process that
regulates arsenic bioavailability. Arsenate
is linked to amorphous Fe and Al oxides,
by the formation of inner sphere surface
complexes, while arsenite forms inner
sphere and outer sphere complexes, the
latter specifically with Al oxides.
In the soil from the area of Gela, the trend
CNR Environment and Health Inter-departmental Project
of arsenic mobility is inverse to that of
mercury. Soil pH determines the negative
potential of mineral surfaces, which
increases with increasing pH. The net effect
is a decrease in sorption processes in the
solid phase of soil. The transport of arsenic
in soil is controlled by sorption/release
processes and the alkaline conditions of
these soils together with the oxidation –
reduction potential, promoting an increase
in the mobility of arsenate ions. Movement
of the contaminant is determined by a pore
space diffusion coupled with a sorption on
solid phases which can be described by a
Freundlich type equation. The solubility of
arsenic can be described by a distribution
coefficient Kd of the divalent arsenate ion
which is directly dependent on soil pH
according to the equation
Log10 Kd = log10 (As soluble/H2AsO- 4) = a
+ b pH
Where Kd is the solid solution distribution
coefficient, for the arsenate ions Assol the
amount released in solution and H2AsO- 4
the free ion activity (127). From this
equation potential bioavailable arsenic
substantially increases with increasing pH.
In the environmental conditions of soils in
the area of Gela there is a high probability
of the existence of soluble arsenic forms.
The soil characteristics, would seem to
indicate that the hazards deriving from this
element could be even higher than those
deriving from mercury contamination.
Environmental issues regarding metals
such as mercury, arsenic (11, 14, 70, 91) are
strictly linked to soil characteristics in that
immobilization or potential bioavailability
is regulated by parameters that are specific
to the soil (pH, clay content, organic
matter, and cation exchange capacity).
These determine the chemical and
physical conditions that may give rise to
precipitation or solubilization resulting
in an increase in bioavailability and/or
leaching, with a danger of polluting the
3.2 Contamination pathways of organic
In an area with a high degree of pollution
such as the Gela site, the main pathways
of contamination affecting the soil are, in
addition to skin contact and direct ingestion
of soil, absorption by roots, transfer to
the edible part of plants, and direct soil
ingestion by animals during grazing.
The absorption by plants of organic
compounds present in soils is influenced
by the physical and chemical properties
of the compound, by the type of soil and
by the characteristics of the plant. It can
occur both by radical absorption and by
subsequent translocation in the aerial
part, both by leaf absorption of volatile
compounds and contaminated dust.
These issues vary in importance depending
on the compound in question. Hydrophobic
substances (PCB) can be absorbed on the
root surface and remain bound to the lipid
of membranes. This can create serious
problems in some species such as carrots,
which have an ectoderm rich in lipids. Plant
absorption is a complex phenomenon based
either on an active process specific to each
compound, or a passive process in which
organic contaminants are transported by
the transpiration water of the plant.
There are several indicators for predicting
the transferability of organic compounds
from the soil to the plant. For example,
compounds with a log Kow between 1
and 2 are those most likely to be moved
by the aerial part of the plants. Substances
with a half-life of less than 10 days will
tend to disappear from the soil before
being absorbed by the plants, while the
most persistent ones can get into the plant
The fate of pollutants in soil
nutrition processes. The most volatile
compounds with Henry’s constant > of 10-4
tend to evaporate from the soil and given
that they are not absorbed by the roots they
may contaminate the plants through the
leaves by volatilization. Of course, this is a
very schematic approach that can be used
for an initial screening of polluted soils in
order to understand what the immediate
dangers are.
The passage of an organic contaminant
from the soil to the food chain can be
described as a series of consecutive
partition reactions between the solid and
liquid phase of the soil, between the soil
solution and the roots, and between the
roots and the aerial part of the plant. This
series of reactions is influenced by the
characteristics of organic compounds, in
particular by the partition coefficient of
octanol/water, so that compounds with a
low Kow value can be moved more easily
into the aerial part of the plants. On the
other hand, substances with a high Kow
value (PAHs, PCBs, PCDD/F) are adsorbed
by the soil and, if partially uptaken by the
plants, they remain in the root system
(2). Generally, these compounds are not
absorbed, but there can be an accumulation
of some compounds in root crops (confined
to the outer parts of the roots that are
removed before consumption).
Some of the more volatile compounds
may enter the leaves, especially through
the stomata, by atmospheric deposition or
by absorption of the molecule in its vapor
state. For semi-volatile compounds with
a high Kow, translocation from the root
system may be minimal, so the absorption
in the vapor state can become an important
source of leaf contamination. Compounds
with a high lipophilic nature and high
volatility may be present with significant
concentrations in the leaves.
Inside the plant, some substances may
be metabolized in a short period, others
(PAHs, PCBs) much less, though they may
be partially degraded at specific sites. For
example, some nitrobenzene compounds
are degraded in the roots, while some
aromatic chlorinated compounds are
metabolized only in the leaves.
Metabolism takes place depending on the
structure of the chemical contaminant
and the type of plant. For example, when
degradation increases, it decreases the
number of chlorine atoms, and the process
is often only partial with the formation of
The amount of halogenated organic
compounds that can be absorbed by the
plant, including hexachlorobenzene,
a contaminant of interest in the Gela
area, depends on water solubility, the
concentration and organic matter content
in the soil. The immediate risks stem from
whether the plant is able to metabolize
or eliminate the compound before being
harvested, and whether the compound is
transferred to the edible part.
Hexachlorobenzene can result from
various industrial processes. It is very
stable and not particularly reactive, since
it is involved in the adsorption phenomena
at the soil surface, which influence the
volatilization and leaching processes as
well as its preponderance to biological and
chemical degradation or uptake by plants.
Since it is a non-ionic compound, it is subject
to the adsorption process involving Van der
Vaals forces, and it is closely linked to the
content of organic matter, particularly in
soils with a low clay content. Unlike other
chlorobenzenes it has a log Kow > 5.3 and
is therefore difficult to assimilate by the
plant since it is substantially immobilized
by adsorption processes in the soil.
The molecular structure is such that the
leaching process should be quite limited.
However, depending on the characteristics
CNR Environment and Health Inter-departmental Project
of the soil texture, the compound may be
found in groundwater, transported through
the larger pores in the soils, or in soils that
have a tendency to form deep shrinkage
The chemical stability of hexachlorobenzene
makes it particularly persistent in soil and
resistant to biodegradation, with a half-life
of more than 1500 days. This compound
has a remarkable permanency in the
atmosphere with the possible formation
of hydroxyl radicals and a half-life of
two years, which could reach the soil as
a result of precipitation and atmospheric
The principal biodegradation mechanism
is oxidation, which leads to the formation
of hydroxylated aromatic compounds,
followed by the breaking of the benzene
Hexachlorobenzene tends to accumulate
in the roots of plants and remains bound to
lipids of the membranes and the cell walls
with less possibility of translocation due to
its low solubility.
The potential toxicity of hexachlorobenzene
for animals is largely linked to the risk
of direct ingestion of soil by animals
during grazing or through fodder feeding.
Hexachlorobenzene is characterized by
a high volatility that can be an important
pollution pathway, through leaf absorption
of fodder and consequently by animals.
3.3 Conclusions
Soil is a complex system that has allowed
life on earth to exist and facilitated the
birth of agriculture. In addition to being
the most important source of essential
nutrients, it is also a source of pollutants
that reach humans through the food chain
and diet (1). Given that soil quality is vitally
important for our health, it is surprising that
this issue has been studied so little by the
scientific community (86). This probably
stems from the fact that identifying and
understanding the mechanisms linking soil
quality and health, through the intake of
agricultural products or processed foods,
requires detailed and multidisciplinary
expertise which is difficult to coordinate
(42, 43, 44, 45, 46).
An innovative solution is to overcome the
compartmentalization of environmental
aspects and consider a continuum that
goes from the presence of a substance in
the soil, to its transfer into the food chain
with the consequent health effects (94). The
main transfer pathways of substances from
soil to humans have been studied almost
exclusively within contaminated sites. It
is assumed in rather simplistic terms, that
there is a direct correlation between the
concentration in the soil of a given element
(or substance) and its absorption by man
(39). However, what really needs to be
investigated is how, in a broader context
the transfer of contaminants from soil to
humans follows quite complex pathways
(76). These pathways are determined by
the chemical and physical nature of soil
(115) characterized by physical, chemical
and biological equilibriums in a multiphase
system that is thermodynamically open.
4.1 CNR Institutes
The Institute of the Ecosystem Studies of
Pisa (CNR-ISE, Pisa) is involved in the
study of the quality of soil and of shallow
and ground waters. This is because
they play an essential role in life cycles,
ecosystems and our quality of life.
The study of soil quality related to human
health has not been studied much by the
scientific community. The understanding
of mechanisms that link soil quality and
human health through food ingestion needs
The fate of pollutants in soil
Figure 2. pH variability in soils sampled at the Gela site.
Figure 3. CEC variability in soils sampled at the Gela site.
coordinated and multidisciplinary skills. which can increase the risk of pathologies
Such mechanisms are not easy to identify is determined on the basis of duration,
and to address to projects involving a frequency and intensity of exposure.
varied skill set.
The transfer of substances from soil to
CNR-ISE’s research is aimed at studying humans follows complex pathways, in
the mechanisms of transport and the relation to the physical and chemical
transformation of contaminants from properties of the soil and to the
sources to soil. The dose of contaminants characteristics of the biological receptor.
needed to determine biological alterations, The transport of contaminants needs to be
CNR Environment and Health Inter-departmental Project
Figure 4. Organic carbon variability in soils sampled at the Gela site.
tackled starting from the highest critical Possible applications of this research
• To identify contaminant pathways in
state areas.
contaminated sites and surrounding
The studies at CNR-ISE focus on the
areas and the effects of pollutants in
effects of chronic exposure to low levels
soil on dietary uptake.
of contaminants, either individual or in
mixtures, and on the consequences of • To evaluate of the transport of
contaminants from sources to target via
exposure to high doses of contaminants
soil-plant system.
naturally present in the environment.
In this particular field CNR-ISE is The Institute of Biophysics of Genoa
carrying out studies on the bioaccessibility (CNR-IBF, Genoa) has a considerable
of contaminants (i.e. heavy metals and experience in electrophysiology and
selenium) in soil in relation to their ion channel biophysics in nervous and
bioavailability. Bioavailability is the endocrine culture cells, investigated
capacity of a contaminant to interact by patch-recording and voltage-clamp
with the biological world and involves the techniques, and intracellular calcium
remediation of contaminated soil using dynamics, studied by fluorescent probes. In
recent years, these skills have been applied
green technologies.
Within this field, the CNR-ISE group has to the study of heavy metal accumulation
organized national congresses on Soil and toxicity in mammalian cells and the
Quality, Food and Health in cooperation modulation of neurotrasmitter-gated ion
with the Institute of Clinical Physiology channels by metal ions in primary neuronal
(CNR-IFC), University of Bari and the cultures and in recombinant receptors
Local Operative Division of Gorizia of expressed in heterologous systems
CRA-RPS. The congresses were funded (frog oocytes and/or mammalian cells).
by the Italian Ministry of Agriculture and The group has additional expertises in
Forestry and the third edition will be held molecular and cellular biology, including
PCR and RT-PCR, in vitro transcription
in 2010.
The fate of pollutants in soil
and the functional expression of wild type
and mutated protein clones in Xenopus
oocytes, cell culture and mammalian cell
transfection. They also study the effect
of acute and chronic treatment with
heavy metals (Pb, Cd, and others) on cell
survival and the maturation of neurons in
culture by functional and viability tests
and apoptosis/necrosis measurements.
Recent work has characterized some Cd
and Pb permeation pathways through the
neuronal membrane and has identified the
location of specific binding sites on the
NMDA receptor channel for Pb and Ni.
The CNR-IBF group in Genoa also
studies the molecular and cellular
astrocyticneuronal interactions in physiological and
pathophysiological conditions. This is done
using calcium imaging, immunoblotting
and electrophysiological techniques.
They research the features and roles of
P2X7 purinoceptors on primary cultures
of neonatal and adult astrocytes, in
secondary cultures stably transfected with
rat P2X7 or expressing truncated P2X7
receptor, cocultures of neuron/glia and on
purified nerve terminals (synaptosomes)
and astroglial fraction (gliasomes).
Polybrominated diphenyl ethers (PBDEs)
are persistent organic pollutants present
in the food chain and in human blood and
milk. Exposure to PBDEs during pregnancy
and lactation leads to signal pathway
alteration and apoptotic neuronal death.
Such events could play different roles
depending on the developmental stage of
the central nervous system. Scarce reports
are available on specific models, allowing
dissection of diverse mechanisms involved
and temporal sequences of prenatal or
neonatal exposure to PBDEs. Relative
contributions of neurons and glia, and
their bi-directional communication in the
control of glutamate synaptic level and in
excitotoxicity triggering and execution,
are not clearly defined. Knowledge of
the efflux mechanism of the excitatory
aminoacids and of how they regulate in the
early and late phases of exposure to PBDEs,
could lead to the possibility of regulating
extracellular excitatory aminoacid levels
in different neonatal PBDEs phases. As
intracellular Ca2+ accumulation seems
to be a prerequisite for neuron damage
cascade, the dampening of Ca2+ influx
through ionotropic glutamate or purinergic
receptors (e.g. P2X7) could significantly
reduce neuronal damage. We therefore
plan to investigate the glutamate efflux
and cellular Ca2+ levels in physiological
conditions and during PBDE exposition
in vitro models of the neonatal and adult
Parallel studies will be conducted to
investigate whether the amino acid release
from astroglial cells can be modulated by
endogenous signaling molecules through
the regulation of swelling-activated Clchannels. The following experimental
models (from the cerebral cortex of
neonatal and 60 days-old rats) will be
used: i) isolated purified nerve terminals
(neuron model) and gliasome (astroglial
fraction unpolluted by nerve endings,
model for astrocytes ex vivo) from the
cerebral cortex of neonatal and adult rats
after food exposition to PBDEs insult. ii)
in vitro cortical neurons and astrocytes.
A functional and pharmacological
characterization will be carried on these
models out of ionotropic glutamate and
purine receptors by studying the “release”
of glutamate (or [3H]D-aspartate),
and intracellular Ca2+ transients with
fluorescence methods (Fura-2).
Possible applications of this research
• Identification and validation of cellular
models (cultured cells) to establish
CNR Environment and Health Inter-departmental Project
significant alternatives to in vivo animal
tests in toxicology.
• Characterization of metal binding
sites on neurotransmitter receptors
and other ion channels for designing
selective ligands to be used in clinical
• Implementation of biosensors to
appraise the bioavailable fraction of
toxic metals and to establish the factor
of correlated biological risk.
Knowledge of the modes for controlling
extracellular glutamate accumulation and
cellular Ca2+ overload and their relationship
with swelling-activated Cl-channels in
neonatal and adult brains exposed to
PBDE insult would contribute to a rational
therapeutic strategy to neuroprotection
with a multipharmacological approach.
In vitro neonatal models, using controlled
experimental conditions and the dissection
of pollutant mediated neurotransmitter
efflux modes are only at a very early stage
of development. Furthermore, little is
known about the control of the excitatory
neurotransmitter efflux from nerve
terminals and the role of neuronal and glial
counterparts in the control of glutamatergic
transmission and of glutamate level at
glutamatergic synapses in the developing
and adult brain.
The Institute of Biophysics of Pisa
(CNR-IBF, Pisa) is involved in the
quantification of prospective toxicity in
aquatic environments in order to evaluate
the relative risk of the introduction of
unknown contaminants. Water organisms
can be contaminated directly or indirectly.
The former occurs by contact or ingestion
of the substance dissolved in water,
whereas the latter happens when the
contaminant is accumulated in the food
chain. Chemicals present in sewage of
industrial or agricultural origin are liable
to contaminate soil, superficial water as
well as groundwater aquifers and thereby
represent a risk for all water use: drinking
water as well as bathing, irrigation and
breeding water.
Contaminants may be of a chemical or
biological origin. The former mainly consist
of substances with slow degradation rates
and which are therefore easily accumulated
in the soil and aquifers and thus in the first
levels of the trophic chain – photosynthetic
organisms, plants and algae. Chemical
residua, such as pesticides, herbicides
and heavy metals, in animal and human
tissues undergo a biological magnification
process. Their toxicity, often consolidated
by a prolonged presence, represents
an important health risk. Among the
contaminants of a biological origin, those
relevant to health issues are algal toxins:
these may be released in the aquatic
environment and have a toxic effect on
Possible applications of this research
The research group aims to create
“early warning” monitoring systems
on marine, fluvial and basin waters
using microspectroscopy and digital
microscopy. In fact, in vivo and in situ
microspectroscopic and microfluorimetric
measures performed on the photosynthetic
compartments of algae present in
waters contaminated by either organic
compounds and/or heavy metals, reveal
the quantitative effects of the pollutants
on the chlorophyll:carotenoid ratio and
on photosynthetic efficiency. All this
information indicates the quality of
the water and which of the microalgae
analysed may be used either as biosensors
or bioremediation, Digital microscopic
measures, on the other hand, obtained with
optic microscopy techniques and image
processing are able to identify and classify
algal species (even to identify a single
occurrence), to determine water quality
The fate of pollutants in soil
and to recognise those species producing
toxins dangerous to human health.
The Institute of Agro-environmental and
Forest Biology of Rome (CNR-IBAF,
Rome) has a mushroom germ plasm bank.
Their research focuses on the:
- collection and characterization of
the wild germ plasm from different
- use of mushrooms for recycling
agricultural and agroindustrial waste;
- degradation of lignocellulosic materials
for animal feeding;
- biotechnologies for environmental
applications of fungi.
To ascertain the quality and the safety of
both the substrates and the fruiting bodies,
analyses on the presence of xenobiotic
substances, in particular the presence
of heavy metals have been performed,.
Moreover, in the field of the alternative
use of mushrooms, technologies have been
developed aimed at:
- obtaining polysaccharides through
extraction in fruitbodies and in mycelia;
fungal polysaccharides, especially
chitin and chitosan, are widely applied
in pharmacopeia, cosmetics, diets and
environmental applications;
- using mycelium biomass for wastewater
depuration. Several batch studies have
been performed to test the ability of
mushrooms to adsorb heavy metals.
Fungal biomass loaded PVA was then
used in columns for the depuration of
water containing heavy metals.
- using
mycoremediation, in the degradation
of phenols, antimicrobial tetracyclines,
polycyclic aromatic hydrocarbons and
heterocyclic compounds.
Possible applications of this research
Mushrooms can be considered as:
a) living organisms or
b) food.
In case a) mushrooms can be used for
mycoremediation in water depuration
or in organic molecule degradation. It
would be interesting to study the role of
polysaccharides present in the cell walls in
heavy metal adsorption and the degradation
of toxic compounds.
In case b) it is important to consider that
mushrooms are able to adsorb heavy metals
in the soil in which they live or in the growth
substrates used for their cultivation. High
concentrations of metals are toxic and
inhibit growth and fructification, but with
different responses for different metals. It
would be interesting to evaluate the dose
that permits mycelium expansion and
carpophore formation but which may still
be dangerous if the mushroom is used as
human food.
The Institute of Methodologies for
Environmental Analysis (CNR-IMAA,
Potenza) has considerable expertise in the
study of mineralogical and geochemical
risks to human health as well as the use
of geo-materials for therapeutic treatments
(such as pelotherapy and pharmacology).
In recent years CNR-IMAA has applied
mineralogical and geochemical information
in order to:
- investigate the presence of potential
toxic elements in waters and rocks
outcropping in some areas at risk and
also to study the mobility of some
chemical elements due to rock-water
interaction processes;
- identify
factors (temperature, water quality,
trace elements, etc.) affecting the
biominerals present in the human
body (in particular in kidney stones
and bones) and their mineralogical and
chemical composition.
Possible applications of this research
• The identification of lithologic
pollution due to rock-water interaction
processes could lead to the production
CNR Environment and Health Inter-departmental Project
of geochemical maps which could be
considered as tools to protect human
• The chemical, mineralogical, petrological and textural study, carried out
with integrated techniques, can be
used to collect useful information on
the processes of neo-formation and the
transformation of both pathological and
non-pathological biominerals present in
the human body (stones, osteoporotic
bones, teeth etc.).
• The methodological approach involved
(epidemiology and geo-environmental
features) may enable information to
be gathered that would be useful for
preventing and treating some diseases.
• The identification of a procedure for
characterizing and highlighting the use
of mineral sources in paleotherapy and
The research of the Institute for the
Dynamics of Environmental Processes,
(CNR-IDPA, Venice) is focused on studying
the environmental processes, especially
the mechanisms of transport and transfer
of organic and inorganic pollutants, both at
local-regional and global levels.
To understand the accumulation and
transfer processes along the trophic web, it
is essential to study the behaviour of trace
elements and persistent organic pollutants
(POPs) at a chemical and biological level.
In addition their chemistry needs to be
considered in concentration terms, as
well as variations in the chemical species
in which the elements can be present.
In order to completely understand the
processes and mechanisms that control
the involvement of metals and organic
compounds at various organisation levels
of the ecosystem and the interaction levels
between the various trace elements, it
is important to closely examine their
absorption and transport along the trophic
web. The interaction of an element with
other parts of a system depends on its
chemical form, so it is very important to
study the speciation of trace elements and
their effects on the interactions between
various biotic or abiotic compartments
of the environment in depth. As for trace
elements, the same can be said for persistent
organic pollutants (POPs), which include
polychlorobiphenyls (PCBs), polyaromatic
hydrocarbons (PAHs), dioxins and
hexachloro-cyclohexanes. In addition
knowledge of the concentrations of the
diverse congeners in the environment plays
a key role towards a better understanding
of the transport along the trophic web and
subsequently the bioconcentration and the
biomagnification in biota.
Several studies have been carried out
in highly polluted areas and in pristine
environments, such as the Venice Lagoon,
the Ross Sea in the Southern Ocean,
Morocco, Vietnam, Mexico, etc. Different
environmental matrices were sampled
(seawater, sediments, biota, etc.) and the
concentrations of organic micropollutants
and of trace elements were assayed;
furthermore, the speciation of trace
elements were studied. It is known that
several organic pollutants, such as PCBs,
can bioaccumulate within the trophic web,
at a level directly related to environmental
levels, and levels within an organism’s diet.
Therefore for an accurate risk assessment,
all the information on congener levels in
the biota and the environment has been
integrated with the WHO Toxic Equivalent
Quantities (TEQs).
The Venice Lagoon represents a particular
ecosystem, a transition between two very
different environments: the Adriatic Sea
and a drainage basin. The variety of inputs
(fresh waters, including run-off from
agricultural soil and contaminated industrial
sites, seawater, industrial and urban wastes
The fate of pollutants in soil
and the input of pollutants via aerosol)
deeply affect the environment, which is
also characterised by a high biodiversity
and high productivity determined by
the input from the drainage basin. Thus,
taking into account the particular features
of this environment, the Venice Lagoon is
among the areas protected by the European
Framework Directive (22nd December
2000, aka Water Framework). According
to the Water framework, in-depth studies
on the environment are required at a
morphological and ecological level. These
should study the impact of environmental
change taking into account any socioeconomic impacts. Of primary importance
are methodological studies to develop
guidelines to evaluate environmental risks
that operate not just at a technical or legal
level but also at a scientific level.
Research also focuses on monitoring
stress biomarkers in indicator organisms
specifically chosen for each environment
under study. The environmental monitoring
of biomarkers and bioindicators could
provide fundamental information. This
would contribute towards an improvement
of direct measurements, such as chemical
measurements by looking at the speciation
of potentially toxic elements. Thus for a
correct evaluation of environmental risks,
analyses need to take a holistic approach.
Biomonitoring and chemical measurements
need to be integrated, taking into account
the diversity and similarities between
organisms and between organisms and
their environment. This would contribute
towards as complete a vision as possible of
all the possible transport routes, and all the
possible exposure and assimilation modes,
as well as bioaccumulation and toxicity
The contamination of waters and sediments
in coastal areas and harbours is due to
a wide range of organic (POPs, such as
PCBs, PAHs, etc.) and inorganic pollutants
(trace elements, such as mercury (Hg), lead
(Pb), chromium (Cr), etc.). In these areas
sediments may be a significant sink and/
or source of these pollutants. Taking into
account the necessity to dredge sediments
in order to keep navigation channels open,
remediation and environmental recovery
are of great consequence in harbour
areas. In fact, the management of dredged
sediments is crucial for the growth of the
port of Venice, due to increasing sea traffic
and foreign trade. In view of the dredging of
many millions of cubic meters of sediment
according to the ‘Piano di Recupero
Morfologico’, CNR-IDPA carried out a
sediments remediation project (RISED,
Azione Biotech III, Regione Veneto),
in collaboration with the Venice Port
Authority. The main aim of this project was
to assess an innovative washing procedure
for dredged sediments. The goal was to
been environmentally friendly and suitable
for the variety of organic and inorganic
pollutants, by exploiting the properties of
natural organic substances.
Activities, future applications and
Analyses of organic micropollutants are
carried out in the CNR-IDPA laboratory,
by gas chromatography coupled with
high resolution mass spectrometry (GCHRMS). In addition to the traditional
extraction systems, there is a pressurised
solvent extraction system (One-PSE,
Applied Separations), an automated sample
purification system (Power-Prep, FMS)
and an automated system for reducing
the sample volume (Turbovap, Zymark)
in the laboratory of the institute. Two
gas chromatographs (HP 6890 Series),
with autosamplers, coupled with EI-MS
detectors (one is a high resolution detector
coupled with a double focus magnetic
CNR Environment and Health Inter-departmental Project
sector Thermo Finnigan Mat XP 95,
the other is a low resolution quadrupole
detector HP 5973) enable several classes of
organic pollutants at trace and ultra trace
levels (PCBs, PAHs, dioxins, emerging
hazardous and/or priority substances, etc.)
to be determined in different and complex
Furthermore, an important future aim is
to establish a risk assessment for POPs
(persistent organic pollutants), which
includes information on toxicity and on the
accessibility and availability towards biota
in a very efficient and pliant way. For a
correct evaluation of environmental risks,
an assay for stress biomarkers, specifically
for the exposure to organic contaminants is
being applied to different species of biota.
A class 100 Clean room minimizes any
contamination of samples to be analysed
for trace elements. Three differently
equipped ICP-MS instruments, namely a
Thermo Finnigan Element 2, coupled with
an autosampler, an Agilent Technologies
7500i coupled with an autosampler, and
an Agilent Technologies 7500cx equipped
with a collision cell, enable the analysis
of elements at trace and ultra trace
concentration levels in different matrices
(seawater, sediments, air, food, different
species of biota). In order to better
understand the bioavailability and the bioaccessibility of trace elements, the study of
geo-speciation in sediments is carried out
using sequential extraction and analysis
by ICP-MS. Since the bioavailability of
trace elements depends on speciation,
it is essential that analytical methods
are available to determine or predict the
bioavailable fraction of a metal. Thus, the
speciation of trace elements such as arsenic
(As) and mercury (Hg) is studied using
various methods. For a correct evaluation
of environmental risks, an assay for the
stress biomarkers for the exposure to
different species of trace elements is being
applied to different species of biota.
The use of new technologies for
environmental monitoring is fundamental
when planning environmental recovery
scenarios in order to appropriately manage
particular environments. The results
obtained in the RISED project were very
promising (both organic and inorganic
pollutants appreciably decreased after the
treatment), due to the holistic approach
used for the various classes of pollutants.
Furthermore, dredged sediments are no
longer harmfully toxic waste, so they
may be a very important resource for
recovering the lagoon landscape. This
study also underlines the importance of
speciation, since according to the most
recent frameworks on risk assessment,
it is essential to know the bioavailability
and bioaccessibility of pollutants in order
to plan the most suitable remediation
project. Future research includes the
possible application of various natural
organic substances and the synergy of
sediment washing with other remediation
techniques, such as bioremediation or
Throughout the world there is a
growing awareness of environmental
management, which is associated with
careful environmental risk management.
The Biotic Ligand Model (BLM) is an
important instrument for assessing the
risk posed by trace elements. It can be
used to determine the bioavailability of
a trace element, and the sensitivity of
organisms to it as a function of its aquatic
chemistry. Thus this a tool for estimating
site specific toxicity factors of the elements
under question. Future research needs to
apply the best “tool-box” according to
the Water Framework, so as to establish
the environmental risk assessment from a
scientific, technical and legislative point of
The fate of pollutants in soil
The Methodological Chemistry Institute
has carried out studies concerning the
performance of chromatographic and
electrophoretic methods for the analysis of
different classes of organic pollutants and
their degradation products in water, soil
and plants and milk. The following classes
of compounds have been considered:
- Pesticides (among which are some
EDCs such as hexachlorocyclohexane,
fenvalerate, linuron, atrazine)
- Phenols
- VOCs ( Benzene, Toluene, Xylene ,
aliphatic ketones and alcohol in goat
In recent bioremediation experiments
soil, plants and rhyzosphere has been
studied, together with the degradation
of such compounds by bacterial strains
inoculated in poplar plants in a greenhouse
The degradation of PAHs and phenols by
ligninolytic fungi and the interaction of
PAH metabolites with humic acids have
also been studied.
Possible applications of this research
Interaction of organic pollutants and
their metabolites with humic matter and
dissolved organic matter in soil and water.
Identification of biodegradation compounds
of organic pollutants in soil by bacteria and
fungi and uptake by plants.
Transfer of VOC from the environment to
mammalian and milk contamination
4.2 External collaborations
The Department of Biology and Chemistry
of Agro-Forestry and Environment at the
University of Bari (DIBCA, Bari) is mainly
involved in various studies related to soil
chemistry and biochemistry, such as:
- the monitoring, conservation and
improvement of soil fertility in order to
maintain soil quality and increase crop
- the role of soil in complex
biogeochemical cycles also disturbed
by anthropogenic impacts (spillings,
amendments, disposal, reclamations,
- the processes and techniques of soil and
sediment decontaminations from heavy
metals and various xenobiotics;
- the physical, chemical and biological
indicators for soil health and quality
in response to internal and external
anthropogenic activities;
- the environmental risks deriving from
the agricultural use of geneticallymodified organisms (GMO).
These objectives are achieved through
a complex theoretical and experimental
chemical, physical and biological
phenomena and processes occurring in
the soil-plant-xenobiotics system. With
given environmental conditions, this
consequently leads to a correct and rational
use of natural soil resources balancing soil
productivity and soil protection.
Possible applications of this research
Monitoring heavy metal and organic
pollutants through the food chain,
especially in the soil-plant system:
relationships between pollutants and soil
organic components and related fractions;
degradation and retention phenomena of
conventional and organic pesticides in
various soil systems.
Evaluation of the physico-chemical
transformation of organic components in
relation to organic waste amendments.
Millions of chemicals are released into the
CNR Environment and Health Inter-departmental Project
environment, and end up in the soil; the
impact of most of them on human health
is still not fully known. There are also
naturally occurring amounts of potentially
toxic substances in the soil whose fate in
the terrestrial environment is still poorly
known. The behavior of contaminants in
soil is related to both the contaminants
and the characteristics of the soil. The
soil properties regulate the distribution of
a substance among the soil phases (solid,
liquid, gaseous) and thus determine the
retention, the release and the migration of
each contaminant (26, 62, 102, 107).
Pollution is also regulated by a time factor,
which influences the availability from the
source emission to the final target. Human
exposure to soil pollution is therefore
time-dependent both directly and through
secondary transfers such as the food
Unlike our knowledge of the exposure
of soil to water and air pollutants, our
understanding of the effects of soil
contamination is still in its early stages, due
to the numerous reactions that take place
in the soil (sorption, release, degradation,
ageing). These reactions modify the
bioavailability of contaminants, which is
dependent on the specific characteristics
of each particular soil. All these reactions
are also influenced by natural weathering
processes, which contribute to the
transport and the erosive migration of
As a result, and despite several legislations
regarding soil, models of pollutant
behaviors are very poorly defined,
especially in terms of the terrestrial food
chain. The transfer of contaminants from
soil to the food chain requires a detailed
knowledge of the complex reactions, that
influence their bioaccessibility and these
reactions are completely different from
those defining the source of exposure.
A study of the fate of contaminants in the
soil can thus provide a reasonable estimate
of exposure. This can then be used as a
basis on which to evaluate adverse effects
on human health (4, 56, 79, 94). There is
still a great uncertainty regarding this issue
and the need to tackle the following issues
has been recognized at an international
- the dietary uptake from vegetables grown
in polluted soils;
- accidental soil ingestion;
- bioaccessibility and bioavailability.
Most attention is now focused on the
last issue, since the bioavailability of
contaminants in soil (65, 71) is quite
different from that deriving from the
toxicological experiments on which the risk
assessment is founded. Therefore only a
deeper understanding of the characteristics
of the soils that regulate the chemical and
biological reactions of contaminants will
contribute to decreasing the uncertainty in
risk assessments (36, 41, 79, 112), and the
consequences of soil pollution on human
One of the main objectives of PIAS-CNR
project “Environment and Health” was
to highlight the close links between
environmental matrices and human
health. Within this framework, Working
Group 1 (WG 1) focused attention on the
fate of contaminants in the environment,
particularly on the soil ecosystem. The
fate of contaminants in the soil depends
not only on the original chemical form
of the contaminants, but also on the
specific characteristics of the soil. The
ability to accurately determine the
effects of contaminants on individual
species, populations, communities and
ecosystems is hampered by an uncertainty
in the quantification of receptor exposure
pathways. Laboratory and field studies
have shown that hazards for human health
did not derive from the total concentration
of a contaminant in the soil, but from the
The fate of pollutants in soil
fraction that is biologically available for that
population at that specific time and with
certain soil conditions. It is now common
knowledge that total concentrations are not
useful to explain the effects of contaminants.
These effects may differ from soil to soil
depending on soil characteristics and
environmental conditions. However one
of the main shortcomings of the present
procedures to evaluate the risks for human
health is the inadequacy, or total absence,
of incorporating the bioavailability of
Half of a century ago, total diet studies were
initiated in response to concerns regarding
the loss of food quality. Nowadays the list
of foods analyzed is continuously updated
and through the database of U.S. Food and
Drug Administration (FDA) it is possible
to know the content of pesticides, heavy
metals, dioxins and other contaminants in
many foods and beverages. These studies
represent a suitable tool for monitoring
dietary intake both in industrial and in
developing countries. Many studies have
also been carried out in Italy and it appears
that most of the elements in the Italian total
diet derived from plant foods (88).
The WG 1 proposal aims to go beyond total
diet studies and to understand mechanisms
and processes by which contaminants
enter the food chain and influence to
various extents nutrition and the health of
The general objective of this proposal
concerns the establishment of a knowledge
network among all the PIAS Working
Groups. This will at the same time
enable each WG to promote and conduct
research strategies in medical, especially
epidemiological circles. However, it is
necessary to devote the same attention
to soil pollution as has been previously
given to air and water pollutions. In this
respect, bioavailability processes assume
an essential role to efficiently describe
the occurrence of contaminants in food,
the effects of soil quality on the quality
of food products, and the implications for
human health.
The working path of the WG 1 Proposal
project starts with the study of contaminant
bioavailability in soil. Bioavailability can
be defined as the degree to which chemicals
present in the soil matrix may be absorbed
or metabolised by human or ecological
receptors or are available for interaction
with biological systems.
In addition, bioavailability depends
on time. Due to ageing, contaminants
binding to the soil may become stronger,
consequently reducing the effects on the
environment. On the other hand, due to
natural or anthropogenic changes in soil
factors (e.g. pH) contaminants may become
more available. The task of soil chemistry
is to define the available fractions, the
potentially available fractions, and the
non-available fractions of contaminants in
different environmental conditions.
The proposal is to study contaminant
bioavailability in three geographical
areas characterized by soils of different
origins where it is possible to find either
areas with a high degree of pollution
due to contamination sources (known as
“Site of National Interest: SIN”), areas
characterized by natural high levels of
metals (Cr, As, Hg), and areas characterized
by the absence of point source pollution.
The general aim of this proposal is to
identify the transfer of contaminants from
soil to the food chain and to evaluate the
possibility that in highly polluted areas,
soil might not be able to exert its essential
role as a filter for ground waters.
The outcome of this part of the project is
also to produce a model or to examine the
possible use of an existing model (such
as the Dutch CSOIL model) to evaluate
the routes through which individuals
CNR Environment and Health Inter-departmental Project
are exposed to a given pollutant. The
main routes to consider are: ingestion of
soil by children, inhalation of soil dust,
and the consumption of vegetables. This
evaluation must be carried out by inserting
bioavailability concepts in the models to
obtain reliable data, which would also be
useful in deriving the concentration of a
contaminant in a specific soil in relation to
human health.
All the research centers mentioned
in Chapter 4 will provide their own
contribution throughout the whole
pathway of the project (Figure 5) under the
coordination of CNR-ISE (Pisa).
The first step is to detect the pollutant
entity (structure, agent) in soil and
water. CNR-ISE (Pisa), will provide an
identification of contaminant pathways
and an evaluation of the transport of
contaminants from sources to target via
soil-plant systems. DIBCA (Bari) will
provide the monitoring of heavy metal and
organic pollutants through the food chain,
as well as the study of the relationships
between pollutants and organic soil
components and related fractions. Through
the identification of geo-environmental
pollution due to rock-water interaction
processes, CNR-IMAA (Potenza) will
create geochemical maps for use as tools
to protect human health.
The second step is to understand
the mechanisms which influence the
transport of chemicals in the soil-plant
system. Using microspectroscopy and
digital microscopy, CNR-IBF (Pisa) will
examine the transport, diffusion and
accumulation processes of pollutants
from the accumulating matrix (soil and
water) to plants and animals. CNR-IDPA
(Venice) will focus on environmental
processes, especially the transport and
transfer of organic (polychlorobyphenils,
polyaromatic hydrocarbons, dioxins and
hexachloro-cyclohexanes) and inorganic
(trace elements, such as mercury, lead,
chromium, etc.) pollutants. CNR-IDPA
(Venice) is involved in the analysis of
organic micropollutants, using gaschromatography coupled with highresolution mass spectrometry and elements
at trace and ultra trace concentration levels
in different matrices with three differently
equipped ICP-MS instruments. CNR-IMC
(Rome) will focus on the analysis of
different classes of organic pollutants
and on the identification of compounds
subjected to biodegradation in soil by
bacteria and fungi, and to uptake by
plants. CNR-IBAF (Rome) investigates
the ability of mushrooms to absorb heavy
metals in soil. Mushrooms can be used
for mycoremediation in water depuration
or in organic molecule degradation.
The third step is the study of pollutant
bioavailability in order to plan the most
suitable remediation strategy. ISE-Pisa will
study the bioaccessibility of contaminants
in soil in relation to their bioavailability.
CNR-IDPA (Venice) will use analytical
methods to determine or predict the
bioavailable fraction of a metal.
The fourth step is an investigation on the
transfer of pollutants from soil to humans
through the food chain. CNR-IBF (Genoa)
will focus on the characterization of
metal binding sites in neurotransmitter
receptors and other ion channels for the
design of selective ligands to be used in
clinical pharmacology. CNR-IBF (Genoa)
will also study the implementation
of biosensors in order to appraise the
bioavailable fraction of toxic elements and
to establish the correlated biological risk
factors. A comprehension of the control
mechanisms of neurotransmitter receptors
and ion channels in the neonatal and adult
brain exposed to a specific class of organic
pollutants (polybrominated diphenyl
The fate of pollutants in soil
Figure 5 – Project proposal
ethers) would allow a rational therapeutic
strategy in a multipharmacological
approach to neuroprotection. CNR-IDPA
(Venice) will focus on the absorption and
transport of metals and organic compounds
along the trophic web. Knowledge of the
concentration of the different congeners
in the environment plays a key role in
biomagnification in biota. The chemical,
mineralogical, petrological and textural
study, carried out with CNR-IMAA
(Potenza) integrated techniques, will
enable useful information to be gathered
on the processes of neo-formation and
transformation of both pathological and
non-pathological biominerals present in
the human body (stones, osteoporotic
bones, teeth etc.). The methodological
approach involved (using epidemiological
and geo-environmental features) may
provide useful information for preventing
and treating some diseases.
All the previous steps will be completed by
an integration of epidemiological studies
carried out by CNR-IFC (Pisa).
With the contribution of WG1 participants:
F. Bianchi (IFC), W. R. L. Cairns (IDPA), P.
Cescon (IDPA), F. Corami (IDPA), L. Cori
(IFC), E. Galli (IBAF), P. Gualtieri (IBF),
C. Marchetti (IBF), T. Miano (Univ. Bari
DIBCA), M. Nobile (IBF), R. Piazza (IDPA),
C. M. Polcaro (IMC), V. Summa (IMAA)
environmental health, heavy
organics, soil quality.
Abrahams P.W. Soils: their implications
to human health. The Science of the Total
Environment 2002; 291: 1-32.
Adriano D.C. Trace elements in the
terrestrial environment. 2nd edn. New
CNR Environment and Health Inter-departmental Project
York: Springer-Verlag, 2001.
Akter KF, Owens G, Davey DE, Naidu
R. Arsenic speciation and toxicity in
biological systems. Rev. Environ. Contam.
Toxicol. 2005; 184:97-149.
Alexander M. Aging, bioavailability, and
overestimation of risk from environmental
pollutants. Environ. Sci. Technol. 2000;
34: 4259-4265.
Armitage J. and Gobas F. A terrestrial
food chain bioaccumulation model
for POPs. Environmental Science and
Technology 2007; 41: 4019-4025.
Ashford NA. and Miller CS. Chemical
exposures: low levels and high stakes.
New York: Van Nostrand Reinhold,
ASTDHPPHE, 2001. Association of
State and Territorial Directors of Health
Promotion and Public Health Education
web-site. Available at: http://www.
ATSDR. Agency for Toxic Substances and
Disease Registry. Toxicological Profile for
Polychlorinated Byphenils (PCBs). http://
Baht RV. and Moy GG. Monitoring
and assessment of dietary exposure to
chemical contamination. World Health
Stat. Q. 1997; 50:132-149.
Baskin LS, Himes L, Colborn T.
Hypospadias and endocrine disruption:
is there a connection? Environ. Health
Perspect. 2001; 109: 1175-1182.
Basu A, Mahata J, Gupta S, Giri A.K.
Genetic toxicology of a paradoxical
human carcinogen, arsenic: a review.
Mutat. Res. 2001; 488: 171-194.
Bell SG. and Todd GA. Detection, analysis
and risk assessment of cyanobacterial
toxins. In: Hester R.E, Harrison R.M,
editors. Agricultural Chemicals and the
Environment. Issues in Environmental
Science and Technology 5. Cambridge:
The Royal Society of Chemistry, 1996. p.
Beyer A. and Biziuk M. Environmental
fate and global distribution of
polychlorinated biphenyls. Rev. Environ.
Contam. Toxicol. 2009; 201: 137-158.
14. Bhattacharya P, Welch H, Stollenwerk
K.G, McLaughlin M.J, Bundschuh J,
Panaullah G. Arsenic in the environment:
Biology and Chemistry. 2007; 379: 109120.
15. Bimbaum LS. and Staskal DF. Brominated
flame retardants: cause for concern?
Environ. Health Perspect. 2004; 112:
16. Black R. Micronutrient deficiency –
an underlying cause of morbidity and
mortality. World Health Organ. 2003; 81
17. Bodar CW, Pronk ME, Sijm DT. The
European Union risk assessment on zinc
and zinc compounds: the process and the
facts. Integr. Environ. Assess. Manage.
2006; 1: 301-319.
18. Boening DW. Ecological effects,
transport, and fate of mercury: a general
review. Chemosphere 2000; 40: 13351351.
19. Boguszeweska A. and Pasternak K.
process of the human organism. Ann.
Univ. Mariae Curie Sklodowska Med.
2004; 59: 524-527.
20. Brady NC. and Weil RR. The nature and
properties of soils. 12th edition. New
Jersey: Prentice Hall, 1999. (881 pp.)
21. Brand E, Otte JPA, Lijzen RIVM. CSOIL:
an exposure for human risk assessment of
soil contamination. A model description.
Rapport 711701054.
22. Bro-Rasmussen F. Contamination by
persistent chemicals in food chain and
human health. Sci. Total Environ. 1996;
188: S45-60.
23. Brunekreef
epidemiology and risk assessment.
Toxicology Letters 2008; 180: 118-122.
24. Carpenter D.O. Polychlorinated biphenyls
and human health. J. Occup. and Med.
Environ. Health 1998; 11: 291-303.
25. CCME. Recommended Canadian Soil
Quality Guidelines. Winnipeg: Canadian
Council of Ministers of the Environment,
1997. (185 pp.)
26. Centers
Environmental Public Health Indicators.
The fate of pollutants in soil
National Center for Environmental Health,
Division of Environmental Hazards and
Health Effects, Atlanta, 2003.
Chee-Sanford JC, Aminov RJ, Krapac
IJ, Garrigues-JeanJean N, Mackie RI.
Occurrence and diversity of tetracycline
resistance genes in lagoons and
groundwater underlying two swine
production facilities. Appl. Environ.
Microbiol. 2001; 67: 1494-1502.
Christensen F.M. Pharmaceuticals in the
environment: a human risk? Regulatory
Toxicology and Pharmacology. 1998; 28:
Colborn T, vom Saal FS, Soto AM.
Developmental effects of endocrinedisrupting chemicals in wildlife and
humans. Environ. Health Perspect. 1993;
101: 378-384.
ComEC (Commission of the European
Communities). Proposal for a Directive
of the European Parliament and of the
Council establishing a framework for the
protection of soil and amending Directive
2004/35/EC. COM(2006)232 final, 22
September. Brussels: Commission of the
European Communities, 2006.
ComEC (Commission of the European
Communities). Thematic strategy for
soil protection. COM (2006) 231 final, 22
September. Brussels: Commission of the
European Communities, 2006.
Communities. A European environment
and health strategy. Coomunication
from the Commission to the Council, the
European Parliament, and the European
Economic and Social Commettee.
[COM (2003) 338 final]. 2003. http://
Cooks JT, Frank DA, Levenson SM et
al. Child food insecurity increases risks
posed by household food insecurity
to young children’s health. Journal of
Nutrition 2006; 136: 1073-1076.
Costa LG, Giordano G, Tagliaferri S,
Caglieri A, Mutti A. Polybrominated
diphenyl ether (PBDE) flame retardants:
environmental contamination, human
body burden and potential adverse health
effects. Acta Biomed. 2008; 79;:172-183.
Crounse RG, Pories WJ, Bray JT, Mauger
RL. Geochemistry and man: health and
disease. 2. Elements possibly essential,
those toxic and others. In: Thornton
I, editor. Applied Environmental
Geochemistry. London: Academic Press,
1983. p. 309-333.
Currie S. Applying the precautionary
principle: an overview. http://www. SNIFFER
(Scotland and Northern Ireland Forum for
Enviromental Research), 2005.
Darnerud PO, Eriksen GS, Johannesson T,
Larsen PB, Viluksela M. Polybrominated
diphenyl ethers: occurrence, dietary
exposure and toxicology. Environ. Health
Perspect. 2001; 109: 49-68.
De Rosa CT, Pohl HR, Williams M,
Ademoyero AA, Chou CHSJ, Jones
DE. Public health implications of
environmental exposures. Environ.
Health Perspect. 1998; 106: 369-378.
de Vries W, Römkens PF, Schütz G.
Critical soil concentrations of cadmium,
lead and mercury in view of health effects
on humans and animals. Rev. Environ.
Contam. Toxicol. 2007; 191: 91-130.
de Wit CA. An overview of brominated
flame retardants in the environment.
Chemosphere. 2002; 46: 583-624.
Dearwent SM, Mumtaz MM, Godfrey
G, Sinks T, Falk H. Health effects of
hazardous waste. Ann. N.Y. Acad. Sci.
2006; 1076: 439-448.
DEFRA & Environment Agency.
Assessment Model (CLEA): Technical
Basis and Algorithms. Department for the
Environment, Food and Rural Affairs and
The Environment Agency, Bristol, 2002
DEFRA (Department for Environment,
Food and Rural Affairs). Contaminants
in soil. Collation of toxicological data and
intake values for humans. CLR9, Bristol,
UK. Department for the Environment,
Food and Rural Affairs and the
Environment Agency, 2002.
DEFRA (Department for Environment,
CNR Environment and Health Inter-departmental Project
Food and Rural Affairs). Sources and
impacts of past, current and future
contamination of soil. Appendix 1: Heavy
Metals. Defra Project Code: SP0547.
London; Defra, 2006.
DEFRA (Department for Environment,
Food and Rural Affairs). Total diet
study – aluminium, arsenic, cadmium,
chromium, copper, lead, mercury, nickel,
selenium, tin and zinc. The Stationary
Office, London, 1999.
contaminated land. A proportionate
approach. Soil guideline values the way
forward. Defra, 2006. http://www.defra.
Di Diego ML, Eggert JA, Pruitt RH,
Larcom L. Unmasking the truth behind
endocrine disrupters. Nurse Pract, 2005;
30: 54-59.
Dickson LC. and Buzik SC. Health risks
of “dioxins”: a review of environmental
and toxicological considerations. Vet.
Hum. Toxicol. 1993; 35: 68-77.
Díez S. Human health effects of
methylmercury exposure. Rev. Environ.
Contam. Toxicol. 2009; 198: 111-132.
Domingo JL. Polychlorinated diphenyl
ethers (PCDEs): environmental levels,
toxicity, and human exposure. A review
of the published literature. Environ Int.
2006; 32: 121-127.
Domingo JL. and Bocio A. Levels of
PCDD/PCDFs and PCBs in edible marine
species and human intake: a literature
review. Environ. Int. 2007; 33: 397-405.
Dudka S and Miller WP. Accumulation
of potentially toxic elements in plants
ant their transfer to human food chain. J.
Environ. Sci. Health. 1999; 34: 681-708.
EPA. Enviromental Protection Agency
Toxic Release Inventory, 2008. http://
Falk-Filipsson A, Hanberg A, Victorin
K, Warholm M, Wallen M. Assessment
factors – applications in health risk
assessment of chemicals. Environmental
Research 2007; 104: 108-127.
Fattore E, Fanelli R, La Vecchia C.
Persistent organic pollutants in food:
public health implications. J. Epidemiol.
Community Health 2002; 56: 831-832.
Ferguson CC. Assessing human health
risks from exposure to contaminated land.
Land Contam. Reclam. 1993; 4: 159-170.
Floyd P. Future perspective s on risk
assessment of chemicals. In Issues in
environment and technology (Vol. 22,
pp. pp. 45-64). London: Royal Society of
Chemistry, 2006.
Food and Nutrition Board, Dietary
reference intakes (DRIs). Recommended
intakes for individuals. Institute of
Medicine, National Academy of Sciences,
Fotakis G. and Timbrell JA. Role of trace
elements in cadmium chloride uptake in
hepatoma cell lines. Toxicol. Lett. 2006;
164: 97-103.
Frederiksen M, Vorkamp K, Thomsen
M, Knudsen L.E. Human internal and
external exposure to PBDEs – a review of
levels and sources. Int. J. Hyg. Environ.
Health 2009; 212: 109-134.
Gaetke LM. and Chow CK, 2003. Copper
toxicity, oxidative stress, and antioxidant
nutrients. Toxicology. 2003; 189;:147-163.
Garelick H, Jones H, Dybowska A,
Valsami-Jones E. Arsenic pollution
sources. Rev. Environ. Contam. Toxicol.
2008; 197: 17-60.
Gilles HM. and Ball PAJ, editors.
Hookworms infections. Amsterdam:
Elsevier 1991. (253 pp.)
Green E, Short SD, Stutt E, Harrison
PTC. Protecting environmental quality
and human health: strategies for
harmonization. Sci. Total Environ. 2000;
256: 205-213.
Grøn C and Andersen L. Human Bioaccessibility of Heavy Metals and
PAH from Soil. Miljøproject Nr. 840,
Milyøministeriet, Copenhagen, Denmark,
Halling-Sorensen B, Nielsen SN, Lanzky
PF, Inger-slev F, Lutzhoft HCH, Jorgensen
SE. Occurrence, fate and effects of
pharmaceuticals substances in the
environment – a review. Chemosphere,
The fate of pollutants in soil
1998; 36: 357-395.
67. Hamilton D, Ambrus A, Dieterle R et al.
Pesticide residues in food: acute dietary
exposure. Pest. Manag. Sci. 2004; 60:
68. Hawley JK. Assessment of health risk
from exposure to contaminated soil. Risk
Anal. 1985; 5: 289-302.
69. He ZL, Yang XE, and Stoffella PJ. Trace
elements in agroecosystems and impacts
on the environment. J. Trace Elem. Med.
Biol. 2005; 19: 125-140.
70. Heikens A, Panaullah GM, Meharg AA.
Arsenic behaviour from groundwater
and soil to crops: impacts to agriculture
and food safety. Rev. Environ. Contam.
Toxicol. 2007 : 189; 43-87.
71. Henschel KP, Wenzel A, Diedrich M,
Fliedner A. Environmental hazard
assessment of pharmaceuticals. Regul.
Toxil. Pharmacol. 1997; 25: 220-225.
72. Hernandez-Ochoa I, Garcia-Vargas
G, Lopez-Carrillo et al. Low lead
environmental exposure alters semen
quality and sperm chromatin condensation
in northern Mexico. Reprod. Toxicol.
2005; 20: 221-228.
73. Hill MJ. Nitrate toxicity: myth or reality?
Br. J. Nutr. 1999; 81: 343-344.
74. Holgate G. The new contaminated land
regime: Part IIA of the Environmental
Protection Act 1990. Land Contamination
and Reclamation 2000; 8: 117-132.
75. Hough RL. Soil and human health:
J. Soil Sci. 2007; 58; 1200-1212.
h t t p : // l n w e b18 .w o r l d b a n k . o r g /
E S S D /e n v e x t . n s f /41 B y d o c N a m e /
76. Hursthouse A. and Kowalczyk G.
Transport and dynamics of toxic
pollutants in the natural environment and
their effect on human health: research
gaps and challenge. Environ. Geochem.
Health 2009; 31: 165-187.
77. Hyams E. Soils and civilization. London:
Murray, 1976. (312 pp.)
78. Hyman M.H. The impact of mercury
on human health and the environment.
Altern. Ther. Health Med. 2004; 10: 70-
79. IRIS. Integrated Risk Information
System-database, US Environmental
Protection Agency, 2003
80. Järup L. Hazards of heavy metal
contamination. Br. Med. Bull. 2003; 68:
81. Järup L, Berglund M, Elinder CG,
Nordberg G, Vahter M. Health Effects
of cadmium exposure – a review of the
literature and a risk estimate. Scandinavian
Journal of Work Environment and Health
1998: 24: 1-51.
82. Johns T. and Duquette M. Detoxification
and mineral supplementation as functions
of geophagy. Am. J. Clin. Nutr. 1991; 53;
83. Jones DL. Potential health risks associated
with the persistence of Escherichia coli
0157 in agricultural environments. Soil
Use Mange. 1999; 15: 76-83.
84. Kazantis G. Mercury exposure and early
effects: an overview. Med. Lav. 2002;
93: 139-147.
85. La Rocca C. and Mantovani A. From
environment to food: the case of PCB.
Ann. Ist. Super. Sanità 2006; 42: 410416.
86. Lee C. Environmental justice: building
a unified vision of health and the
environment. Environ. Health Perspect.
2002; 110 (Suppl. 2): 141-144.
87. Liem AK, Fürst P, Rappe C. Exposure
of populations to dioxins and related
compounds. Food Addit. Compounds
2000; 17: 241-259.
88. Lombardi-Boccia
Cappelloni M, Di Lullo G, Lucarini M.
Total diet study: daily intakes of minerals
and trace elements in Italy. British J. Nutr.
2002; 90: 1117-1121.
89. Luthy RG. Bioavailability of Contaminants
in Soils and Sediments: Processes,
Tools, and Applications. (Ed. National
Research Council US, Committee on
Bioavailability of Contaminants in Soils
and Sediment), The National Academies
Press, Washington, DC, USA, 2003.
90. Mahaffey KR. Methymercury: a new look
at the risks. Public Health Rep. 1999; 114:
CNR Environment and Health Inter-departmental Project
396-399; 402-413.
91. Mandal BK. and Suzuki KT. Arsenic
around the world: a review. Talanta, 2002;
58: 201-235.
92. McArthur JM, Ravenscroft P, Safiulla S,
Thirlwall M.F. Arsenic in groundwater:
testing pollution mechanisms for
sedimentary aquifers in Bangladesh.
Water Resou. Res. 2001; 37: 109-117.
93. McKinlay R, Plant JA, Bell JNB.
Calculating human exposure to endocrine
disrupting pesticides via agricultural
and non-agricultural exposure routes.
Sciences of the Total Environment 2008;
398: 1-12.
94. McLaren L, and Hawe P. Ecological
perspectives in health research. J.
Epidemiol. Community Health 200; 59;
95. McMichael AJ, and Beaglehole R. The
changing global context of public health.
Lancet 2000; 356: 459-499.
96. Mielke HW, Gonzales C.R, Smith MK,
Mielke PV. The urban environment and
children’s health: soils as an integrator of
lead, zinc and cadmium in New Orleans,
Louisiana. USA Environ. Res. 1999; 81 :
97. Millis PR, Ramsey PH, John EA.
Heterogeneity of cadmium concentration
in soil as a source of uncertainty in plant
uptake and its implications for human
health risk assessment. Sci. Total Environ.
2004; 326: 49-53.
98. Morris G, and Robertson R. Environmental health and the health improvement
challenge: a report commissioned by the
Royal Environmental Health Institute of
Scotland. Royal Environmental Health
Institute of Scotland, 2003.
99. Morris G. Determining priorities
in developing and delivering future
environment health services. Environ.
Health Int. 2002; 4: 10-14.
100. Morris GP, Beck SA, Hanlon P, Robertson
P. Getting strategic about the environment
and health. Public Health 2006; 120: 889907.
101. NAS. National Academy of Sciences.
Toxicological effects of methylmercury.
Washington (DC), 2000.
102. National Center for Environmental
Health. National Report on Human
Exposure to Environmental Chemicals.
CDC, 2003. Available at (http://www.
103. Needleman C. Applied epidemiology
and environmental health: emerging
controversies. Am. J. Infect. Control.
1997; 25: 262-274.
104. NEPI. Assessing the Bioavailability
of Metals in Soils for Use in Human
Health Risk Assessments (Ed: National
Environmental Policy Institute, NEPI),
Washington, DC, USA, 2000.
105. Nickson R, McArthur J, Burgess W,
Ahmed KM. Arsenic poisoning of
Bangladesh groundwater. Nature. 1998;
395: 338
106. Northridge ME, Stover GN, Rosenthal
JE, Sherard D. Environmental equity and
health: understanding complexity and
moving forward. Am. J. Public Health
2003; 93: 209-214.
107. O’Neill MS, Jerrett M, Kawachi I, Levy
JL, Cohen AJ. Health, wealth, and air
pollution: advancing theory and methods.
Environ. Health Perspect. 2003; 111;
108. Oliver MA. Soil and human health: a
review. European Journal of Soil Science
1997; 48: 573-592.
109. Olsson IM, Eriksson J, Oborn I,
Skerfving S, Oskarsson A. Cadmium in
food production systems: a health risk for
sensitive population groups. Ambio. 2005;
34: 344-351.
110. Organization for Economic Co-operation
and Development. OECD environment
programme 2005-2006. Organization
Development, 2005. http://www.oecd.
111. Pan J, Plant JA, Voulvoulis N, Oates CJ,
Ihlenfeld C. Cadmium levels in Europe:
implications for human health. Environ.
Geochem. Health 2009; 172: 1145-1149.
112. Paustenbach DJ. Human and ecological
risk assessment: theory and practice. New
York: John Wiley and Sons, 2002.
The fate of pollutants in soil
113. Pearce F. The cause of reef health
problems may be blowing in the wind.
New. Sci. 1999; 163; 22.
114. Pearce F. Farmers’ free for all: Europe
loosens curbs on animal drugs in the soil.
New Sci. 2000; 165: 20.
115. Pedron F and Petruzzelli G. L’influenza
delle caratteristiche dei suoli sulla mobilità
dei contaminanti e il passaggio nella catena
alimentare. Epidemiologia&prevenzione
2009; 33: 45-56.
116. Petruzzelli G and Pedron F. 2007.
Meccanismi di biodisponibilità nel suolo
di contaminanti ambientali persistenti. In:
Comba.P, Bianchi F, Iavarone I, Pirastu R.
(Ed) Impatto sulla salute dei siti inquinati
metodi e strumenti per la ricerca e le
valutazioni . Roma: Istituto Superiore di
Sanità; 2007 (Rapporti ISTISAN 07/50)
117. Pohl H, DeRosa C, Holler J. Public health
assessment for dioxins exposure from
soil. Chemosphere 1995; 95: 2437-2454.
118. Pollitt F. Polychlorinated dibenzodioxins
and polychlorinated dibenzofurans.
Regul. Toxicol. Pharmacol. 1999; 30:
119. Price EW. Non-filarial elephantiasis –
confirmed as a geo-chemical disease,
and renamed “podoconiosis”. Trop. Doct.
1988; 26; 151-153.
120. Ritter WF. Pesticide contamination of
groundwater in the United States – a
review. J. Environ. Sci. Health B. 1990;
25: 1-29.
121. Robson M. Methodologies for assessing
exposure to metals: human host factors.
Ecotoxicol. Environ. Saf. 2003; 56 : 104109.
122. Rose JB. Emerging issues for the
microbiology of drinking water. Water
Eng. Manage. July. 1990; 23: 26-29.
123. Rupert LH, Neil B, Scott DY et al.
Assessing potential risk of heavy metal
exposure from consumption of homeproduced vegetables by urban populations.
Environ. Health Perspect. 2004; 112: 215221.
124. Schlatter C. Environmental pollution and
human health. Sci. Total Environ. 1994;
143: 93-101.
125. Schmidt CW. The lowdown on low-dose
endocrine disrupters. Environ. Health
Perspect. 2001; 109.
126. Sharma RK. and Agrawal M. Biological
effects of heavy metals: an overview. J.
Environ. Biol. 2005; 26 : 301-313.
127. Shelmerdine PA, Black CR, McGrath SP,
Young .D. Modelling phytoremediation
by the iperaccumulating fern, Pteris
vittata, of soils historically contaminated
with arsenic. Environ. Pollution. 2009;
157: 1589-1596.
128. Sridhara Chary N, Kamala CT, Suman
Raj S.D. Assessing risk of heavy metals
from consuming food grown on sewage
irrigated soils and food chain transfer.
Ecotoxicology and Food Safety 2008;
129. Steinemann A. Human exposure, health
hazards, and environmental regulations.
Environ. Impact Assess. Review 2004;
24: 695-710.
130. Tchounwou PB, Ayensu WK, Ninashvili
N, Sutton D. Environmental exposure
to mercury and its toxipathologic
implications for human health. Environ.
Toxicol. 2003; 18: 149-175.
131. Thornton I and Webb JS. Geochemistry
and health in the United Kingdom. Phil.
Trans. R. Soc. Lond. B. 1979; 288: 151168.
132. UNEP, UNICEF, WHO, Children in the
new millennium: environmental impact
on health. 2002.
133. US Department of Health and Human
Services, 2004. Healthy People 2010.
134. US Environmental Protection Agency.
Draft Report on the Environment,
2004. Available at:
135. US Environmental Protection Agency.
Particle pollution and your health, 2005.
Available at:
136. USGS. United States Geological Survey
web-site. Available at: http://water.usgs.
CNR Environment and Health Inter-departmental Project
137. Wagner JC. The pneumoconioses due to
mineral dusts. J. Geol. Soc. Lond. 1980;
137: 537-545.
138. Wilcox BA. Ecosystem health in
proactive: emerging areas of application
in environment and human health.
Ecosystem Health 2001; 7: 317-14.
139. Woodruff TJ, Axelrad DA, Kyle AD.
America’s children and the environment:
measures for contaminants, exposures
and diseases. EPA 240-R-03-2001. US
Environmental Protection Agency, Office
of Policy, Economics and Innovation and
Office of Children’s Health Protection,
Washington, DC, 2003.
140. World Bank. Making sustainable
commitments: an environmental strategy
for the World Bank. World Bank, 2001.
141. World Health Organization. Development
of environment and health indicators of
European Union countries: results of a
pilot study. World Health Organization
Regional Office for Europe, 2004. http://
142. World Health Organization. Environmental health indicators for Europe: a
pilot indicator-based report. www.euro.
World Health Organization Regional
Office for Europe. 2004.
143. World Health Organization. Health
and the environment in the WHO
European region: situation and policy
at the beginning of the 21th century.
EUR/04/5946267/BD/5 World Health
Organization, 2004. http://www.euro.
Water and Soil Monitoring for the Protection
of Environment and Human Health
M. Rusconi, S. Polesello
CNR, Water Research Institute (IRSA), Milan, Italy
[email protected]
The first pillar of the protection of the environment and also, as a positive consequence, of the human
community living in this environment, is the establishment of protective monitoring programs. Current
monitoring programs for water and soil are based on sampling and laboratory analysis of chemical and
microbiological variables. Parallel to this traditional approach, methods to measure effects directly on living
organisms, at both individual and community level, have been integrated into monitoring plans. The present
paper reviews the state of the art of the research activities in the field of water and soil monitoring carried out
by the Italian National Research Council (CNR) Institutes: this review is the outcome of a survey conducted
by the Working Group 2 established in the framework of the CNR Environment and Health Inter-departmental
Project , PIAS CNR. Emerging problems, such as the presence of nanoparticles and perfluorinated compounds
in the environment, and future developments of the monitoring techniques are also discussed.
The protection of the environment
and human health from toxic agents is
traditionally based on the selection of a
list of potentially dangerous substances
or agents and the statement of the
corresponding emission limit values or
quality standards.
Methodologies to establish quality
standards are based on physicochemical
and toxicological data which are normally
collected and organized in a risk assessment
document. The prioritization procedure
used to establish the list of pollutants is
based on the knowledge of the toxicological
properties and data on use and production
amounts. In order to be validated, all these
procedures need a large amount of data
which are actually not available for the
millions of synthetic molecules prepared
and brought into the market.
As a consequence, the approach, based on
emission limit values at the discharge and
on environmental quality standards in the
receptor compartment, could not be really
protective for the environment.
A step forward in the legislation on water
quality protection was the publication of
the Water Framework Directive (WFD,
Directive 2000/60/EC) that introduced
the concept of water bodies protection:
it moved from a regulation based on
emission control to one that is based on
the protection of the ecological quality of
receiving water bodies. On the assumption
that the repression of discharge by
imposing emission limit is not sufficient
to protect the environment, it is crucial to
verify whether the receiving body is able
to support activities which are imposed on
it, keeping as much intact as possible the
ecological community that resides there.
The aim of the WFD is to achieve a good
quality status from the ecological point
of view, namely to ensure that all bodies
CNR Environment and Health Inter-departmental Project
could support a biodiverse ecological
In parallel with this innovative approach
based on ecological classification, in order
to control the chemical pollution, WFD
establishes a priority list of compounds
which, for production volumes and/or use
and hazardous characteristics in terms of
toxicity, persistence and bioaccumulation,
pose a risk to the aquatic ecosystem or
to human health. The pollution control
is based on a combined approach which
sets limits on emissions (left to Member
States) and maximum allowable levels in
the receiving water body, expressed as
environmental quality standards (EQS)
which are fixed in a recently adopted EC
Directive (105/2008/EC).
The use of living organisms and their
community as monitoring tools has many
advantages. Organisms, living in the
environment under study, are constantly
exposed to the physical, biological and
chemical influences of that environment.
significant quantities of compounds even
if exposed to very low concentrations in
the environment.
It is nevertheless difficult to correlate
the measured adverse effects on the
ecological community with the presence
of specific classes of chemical compounds
because also the hydromorphological and
physicochemical alteration of the natural
habitat influences the structure of the
community. In order to detect sub-lethal
effects, single living organisms are the
best indicators of environmental alteration:
if chemical tests only detect “known”
substances, the measuring of effects on
organisms by appropriate biomarkers can
highlight not only the biological response to
unknown substances but can also evidence
the synergistic effects that may be caused
by a mix of different substances.
Integrated monitoring systems are the most
effective tool to highlight the interactions
among substances by pointing out the
responses at different levels: responses
which can be so lethal that they affect the
composition of the community, or sub
-lethal responses that act on the bodymetabolic-physiological models or interact
with genetic transmission mechanisms.
Low-cost or highly innovative technologies
have been developed for the application in
field/in situ (spectroscopic or sensing), in
order to achieve a rapid characterization
of the site with a high spatial/temporal
resolution; in alternative to chemical or
physicochemical monitoring systems,
techniques based on the measurement of
biological response have been proposed,
as they can evaluate the possible risk,
even in the absence of a direct chemical
measurement of a particular pollutant. The
role of biomarkers, possibly integrated
in a biosensor system, is crucial for the
development of an early warning system
which could prevent adverse effects on the
ecological community and human health.
The working group 2 (WG2) on “Monitoring
Systems for Soil and Water”, established
in the framework of the PIAS-CNR
Project, has defined its field of interest in
the development and use of monitoring
techniques, technologies or innovative
methods for soil and water monitoring,
where a situation of environmental
pollution represents a potential risk for
human health.
After a preliminary review of literature
focused on the various techniques
developed for the monitoring of soils and
waters, WG2 researchers were invited
Water and Soil Monitoring for the Protection of Environment and Human Health
to put together their scientific expertise
and produce a state of the art report of
the activities regarding the monitoring of
environmental impacts which could be
relevant for human health. Assuming the
central role of the integrated approach,
a review of the different professional
profiles present in the CNR Institutes was
carried out, to facilitate the creation of a
multidisciplinary group of researchers and
the sharing of different expertise ranging
from instrumental analytical chemistry to
ecological assessment.
Ecological risk assessment (ERA)
has been defined as ‘‘the practice of
determining the nature and likelihood
of effects of anthropogenic actions on
animals, plants, and the environment’’
(1). A correct analysis of the complex
interactions between the pollution caused
by humans and the environment requires
the application of a multidisciplinary
approach and the determination of different
parameters, that can describe the exposure
levels and convert them into individual
warning situations (2).
A clear example of this operating method
is described by the Triad approach used in
the ecological risk assessment of sediments
(2,3) and allows the investigation of the
possible negative effects of toxic chemicals
at different levels of biological organization,
from single organism to population and/or
community level (2). The Triad paradigm
enables the assessment of potentially
hazardous effects on ecosystems by
simultaneously considering chemical
concentration, bioavailability of pollutants
and the ecotoxicological profile of the
environmental matrix under observation.
The latter is usually determined by a set
of ecotoxicological tests as well as by
monitoring possible ecological alterations,
quantified by changes in different structural
and functional community attributes. This
integrated approach on different levels of
monitoring should be adopted to provide
full details of the impact on both the water
and the soil in the site of interest. Methods
that are relevant at different levels of
specificity are needed because some
parameters (such as biomarkers) describe
effects at suborganism levels of biological
organization (4) and different phases
of stress syndrome evolution in model
organisms (5,6). On the contrary, other
ecotoxicological endpoints (e.g. survival
and reproduction) indicate possible direct
effects at population level (7). Chemical
analyses reveal the presence of potentially
dangerous substances in soils, but cannot
be used to quantify bioavailable fractions
(8) that play a more relevant role in
threatening ecosystem integrity (9-11).
Finally, a direct evaluation of community
structure and functionality should clearly
detect overall environmental effects on
ecosystems (12) .
As everybody knows, biomarkers have
been defined as sublethal responses to
environmental chemicals at different levels
of biological organization (e.g, molecular,
cellular, tissutal, physiological, behavioral,
of organisms) which evidence an alteration
respect to the natural status (13,14).
Toxic effects caused by exposure to
environmental pollutants can alter
endpoints at different levels of biological
organization (4,15) (Fig. 1).
In particular, the classical biological
tools applied in ERA (i.e, bioassays and
ecological parameters) are able to highlight
the impairments from the organism to
the population–community level. This
analytical system cannot, however, be used
to investigate early effects in organisms
exposed to pollutants (i.e, from early
CNR Environment and Health Inter-departmental Project
Figure 1: Different biological toxic effects
From Dagnino et al. (15).
sublethal stress syndrome to the onset of
reduced survival). The investigation of
the initial phases of biological impairment
plays a crucial role in determining the
vulnerability level reached by the biotic
resources in those cases where no evident
changes in the traditional, high level
endpoints are detected. Therefore, in order
to complete the analysis of the spectrum
of possible biological effects induced
by environmental contamination, the
alterations in sublethal endpoints measured
on sentinel organisms can be evaluated
(6). In spite of the high sensitivity of these
types of parameters, it must be stressed
that it is possible to clearly infer the stress
syndrome degree in organisms exposed to
toxic chemicals by the application of ad hoc
models. This is done by using a complete
battery of biomarkers at different levels
of biological organization (i.e, molecule,
cell, tissue, organ, organism) on model
organisms (5) .
Therefore an extensive monitoring requires
induced by environmental contamination.
different skills, ranging from purely
analytical capacities for the identification
of compounds present in the matrix
under investigation, up to bio-molecular
techniques to assess effects of pollutants
on the gene expression.
In the following paragraphs, a survey on
the activities of CNR institutes will be
presented, covering many of the skills
described above and needed for the
establishment of an effective monitoring
project. The covered field of expertise
ranges from advanced chemical analysis,
in laboratory as well as field research,
to traditional ecotoxicological assays,
biomarker assessment in exposed and
natural organisms, up to studies about the
alterations of structure and function of
ecological communities.
3.1 Laboratory chemical analysis
The off-site instrumental techniques
are characterized by manual sample
collection and transfer in centralized
Water and Soil Monitoring for the Protection of Environment and Human Health
laboratory units where advanced analytical
equipment is available. Many chemical
monitoring activities with instrumental
methods are currently on-going in CNR
Institutes, also in response to the Water
Framework Directive (WFD, 2000/60/EC)
Several groups are involved in monitoring
metals in Italian soil, surface and
groundwater. Metals present in soil and
waters can accumulate in the trophic chain
and represent a risk for the final consumers.
The Tuscany region in Central Italy, due
to geological and historical reasons (past
mining activities present in the territory
since the Etruscan era), is particularly
impacted by metals and many CNR groups
are working on metals contamination in
this area.
Trace metal concentrations were monitored
in some urban soils of three medium
sized towns of coastal Tuscany. (16) Soil
samples were collected in roadsides, urban
agricultural soils (allotments), playgrounds
and public parks. The analysis included
total metal content (Pb, Cu, Zn, Ni, Cd),
and sequential extraction. Lead reached the
highest levels in the soils and was higher
near roads. In urban agricultural soils and
in allotments Cu was present in noticeable
quantities (300 mg/kg). The presence of Cu
in urban soils seems to be typical of soils
used for a long period as agricultural land,
especially vineyards in the area covered
by this study. Sequential extractions were
performed to evaluate the mobility of the
metals and to better understand the impact
of the anthropogenic activity on urban
Mercury contamination in the Cecina river
basin (Tuscany, Italy) has been studied by
Scerbo et al. (17). Mercury was measured
in waters, sediments and fish of the river
and its most important tributaries. In fish
samples the organometallic metabolites of
mercury were also determined. Particularly
high concentrations were found in the
sediments of the S. Marta channel flowing
into the Cecina, where a chloro-alkali
plant discharges its wastes, and high levels
were still detectable 31 km downstream
from the confluence, bioaccumulating also
in fish species.
Italy, and particularly Tuscany, is strongly
interested by boron contamination
because of the presence in its territory of
active volcanism, geothermal activity and
mineralized areas. The compliance with
the EU normative is technically complex
and economically very expensive. The
limit of 1 mg/l imposed by the European
Union for boron in drinkable waters
(98/93/EC) is based on the “precautionary
principle”, considered that the effect of
boron on humans is at present poor and
investigations on waters (δD, δ O, δ11B,
δ13C, δ15N, 87Sr/87Sr) and soils (δ11B,
Sr/87Sr) were carried out in Southern
Tuscany where boron anomalies occur and
the assumption of this element through
drinking water or agricultural products
could have an adverse effect on the health
of the local population (18-21).
concentrations of trace metals in soil is
crucial to highlight the possible presence
of contamination. These abnormalities are
identified on the basis of the knowledge of
natural concentrations, expressed in terms
of “background” (natural background
levels) or “baseline” (currently found
contents). The criteria for the determination
of these “natural” concentrations have
been the subject of intense international
debate for many years. It is therefore
necessary to evaluate the potential of other
pollution indicators from diffuse sources,
complementary to the existing ones (soils
CNR Environment and Health Inter-departmental Project
and sediments). In order to achieve this,
the properly standardized use of higher
plants offers a promising tool to establish
a precise date for the event and to assess
the spatial extension of the contamination
In parallel to soil and groundwater studies,
presence, distribution and accumulation
of metals in sea areas were investigated:
samples of Mytilus galloprovincialis
were collected monthly during the July
1999-June 2000 period from two mussel
culture areas influenced by urban and
industrial wastes (26). These stations,
subject to different environmental impact
conditions, are located in the coastal area
of Taranto Gulf (Ionian Sea, Southern
Italy). Metals (Cd, Cu, Pb, Zn, Fe and As)
were determined by atomic absorption
spectrophotometry (AAS) in the whole
soft tissue of mussels. Seasonal changes
in metal concentrations were observed.
Metals exhibited maximum values in
later winter-early spring, followed by a
progressive decrease during summer.
Metal concentrations were similar to
those detected in other Italian coastal
zones, and indicate that the seafood under
investigation poses no hazard to human
health because metal content is within
the permissible range established for safe
consumption by humans.
For many years the presence of toxic
inorganic fibrous particles such as asbestos
in drinking waters has been of great
concern for their direct impact on human
The assessment of the diffusion of inorganic
fibrous particles in the environment is
performed through detection, identification
and quantification of mineral inorganic
particles present in animal and human
tissues and biological fluids from impacted
areas. In these same areas, a comparison is
carried out among the types and amounts
of particles detected in the biological
samples and the fibers present in aerosol
and mineral outcrop. An evaluation of the
biological effects of some of the fibrous
mineral phases present in the rocks is also
performed (27-30).
In the field of the advanced methodologies
for the monitoring of organic micropollutants in water, the CNR Water
Research Institute (CNR-IRSA) plays
a well acknowledged role. Besides the
determination on “classical” persistent and
priority organic pollutants (chlorinated
pesticides, PCB, PAH, alkylphenols),
advanced analytical methods based on
LC-MS technique are under development
for the determination of metabolites and
emerging pollutants in various surface,
drinking and ground waters in Italy (3133).
Emerging substances are those compounds
or groups of compounds produced or
used in significant quantities but which
are not currently restricted by regulation
due to the lack of information about their
effective environmental diffusion and
toxicity. In this category, many substances
are polar substances such as perfluorinated
compounds, PPCP (pharmaceutical and
personal care products), estrogens. The
CNR-IRSA research group is also the
national focal point of an international
network, NORMAN (Network of Reference
Laboratories for Emerging substances)
and the one responsible for the substance
prioritization procedures at European level
under the Water Framework Directive
Common Implementation Strategy.
3.2 Biomarker-bioassay
Bioassays are typically used to measure
the effects of some substance on a living
They can be classified on the basis of
the type of response, by discriminating
Water and Soil Monitoring for the Protection of Environment and Human Health
the end-point level: a high response
level is associated to the survival rate
or reproduction inhibition, while a low
response level is connected to sublethal
effects that are revealed by specific
physiological or genomic alterations.
Among the former type, the most
used ecotoxicological tests are those
based on the measurement of EC50
(Effect Concentration) or LC50 (Lethal
Concentration), i.e. the concentrations
which exert an effect on 50% of the
organisms under test. The latter type of
bioassays is based on the measurement
of specific biomarkers: experimental tests
evaluate effects that are not lethal for the
organism, like a change in a metabolic
protein or a behavioral modification.
As indicated by Dagnino et al. (15), the
separate evaluation of different levels of
response helps to avoid a misinterpretation
of the ecotoxicological results.
At the CNR Marine Science Institute
(CNR-ISMAR), studies are carried out
about the health condition assessment of
marine invertebrates and vertebrates in
relation to environmental stressors. (3436) Moreover, there is a great interest
about the deployment of biochemical,
histo-cytochemical and histopathological
biomarkers as early warning systems in
coastal marine environments monitoring
that is the object of the research at the CNR
Institute of Biomedicine and Molecular
Immunology (CNR-IBIM) Cell Stress
and Environment Research Unit (37-46).
This research group has contributed to
the identification of cellular and molecular
stress markers in the marine environment
by using the sea urchin as a model system.
This is a common organism in our shores
and has a great ecological and commercial
importance. Laboratory as well as
field experiments showed that, when
responding to chemical (heavy/essential
metals) or physical (temperature, acidity,
ionizing radiations) stressors, the adult
immuno-competent cells and embryos or
larvae of the sea urchin express specific
markers, better known as stress proteins,
including heat shock proteins (hsp70)
(39-41), metallothionein, (38) and acetylcholinesterase (37). Other studies have
shown that environmental stress causes
DNA damage in the form of broken singleand double-strand (44), and variations in the
levels of other stress and apoptotic markers
in response to exposures to heavy/essential
metals (cadmium/manganese) and/or UVB/X radiation (42,43). Results at cellular
and molecular (proteins and mRNA)
levels from laboratory exposures were
compared to those obtained using samples
collected in field studies carried out in
the Mediterranean and Northern seas, in
order to bridge together field ecology and
laboratory-oriented molecular toxicology
(45,46). Most of the markers tested were
sensitive to the stress conditions used.
The results of the research support the
suitability of sea urchin cells and embryos
as valid tools to bio-monitor the effects of
physical and chemical stress on marine
aquatic ecosystems.
At the CNR Biophysics Institute
(CNR-IBF) studies on the effects of
environmental pollutants in eukaryotic
microorganisms are ongoing; specific type
of physiological, cellular and molecular
level responses have been identified in
the presence of environmental pollutants
(47-52). This kind of studies is a valuable
complement to chemical analysis, as it
can provide valuable information on the
potential toxicity to the organs, in order to
detect the first symptoms of exposure. The
effects studied include:
- the change in the intracellular pool
of non-protein thiols (glutathione and
phytochelatins) in phytoplankton algae
CNR Environment and Health Inter-departmental Project
to be used as a biomarker of heavy
metals bioavailability.
- the variation of photosynthetic
microorganisms in aquatic and
motility in response to exposure to
environmental pollutants.
- short-term genotoxicity tests on cell
cultures of specific strains of yeast used
as a model system.
An integrated approach of biological assays
performed with different microorganisms
can be applied to water and sediment
elutriate collected in impacted coastal
and inland areas, such as estuaries, ports,
urban areas.
In vivo measures by micro-spectroscopy
and micro-spectrofluorimetry of the
photosynthetic compartment of planktonic
species, present in water contaminated
by organic compounds and/or heavy
metals, can determine the effects of
these contaminants on the composition
of pigments, the ratio chlorophyll:
carotenoids, photosynthetic efficiency
(53-56). Toxic algal species identification
and quantification could be done using
techniques of optical microscopy, and/or
fluorescence for taxonomic recognition
with image processing techniques.
The Venice Lagoon (Northern Italy) is an
attractive area of study due to its historical
interest and ecological fragility. A spatial
and temporal survey at three sites located
in the “canals” of Venice city centre and
at a reference site in the Lagoon was
undertaken to evaluate stress effects on
mussels sampled in Venice urban area,
where raw sewage is discharged without
treatment (57). A battery of biomarkers
(metallothionein, micronuclei, condition
index and survival in air) was used
to evaluate the stress condition of the
animals. At the same time, an alkali-labile
phosphate assay (ALP) was performed in
mussel hemolymph to find a biomarker
of the estrogenic effect for this species.
Biomarker results showed an impairment
of the general health condition in the
mussels coming from the urban area, in
agreement with the chemical analysis.
Another study (58) surveyed the water
quality in Venice urban areas in connection
with the discharge of untreated sewage
directly into canals, in addition to the
pollutant load already present in these
areas. One way of estimating the impact
of these chemicals is the monitoring of the
local fauna. In the search for good water
quality indicators in Venice urban area,
two physiological indices for mussels
(Mytilus galloprovincialis) - survival
in air and condition index - have been
evaluated. Native mussels and also those
transplanted into the urban area showed
reduced survivability in air and decreased
condition index values, indicating a less
healthy status in animals collected from
the urban canals. Data are discussed in
relation with pollutant bioaccumulation.
Coastal environments are highly variable
on a daily scale. In these environments,
benthic foraminifera, a class of marine
Protista, can be used as bioindicator (59).
These organisms can define the extent of
similar environmental conditions (biotopes)
through the study of the structure of their
assemblage (presence – absence - relative
dominance). The monitoring results show
the capacity of the benthic foraminifera
to monitor the changes occurring in
unstable environments and to indicate
the evolutionary trends of transitional
The use of molecular techniques such as
PCR assay (Polimerase Chain Reaction)
for the determination of the genotoxic
effects induced by the pollutants in the
monitoring programs is rather recent.
This tool was employed in an assessment
Water and Soil Monitoring for the Protection of Environment and Human Health
study on recharged aquifers (60): this
practice presents advantages for integrated
water management in the anthropogenic
cycle, namely, advanced treatment of
reclaimed water and additional dilution
of pollutants due to mixing with natural
groundwater. Nevertheless, this practice
represents a health and environmental
hazard because of the presence of
chemical contaminants. In this study, the
groundwater recharge systems in Torreele,
Belgium, Sabadell, Spain, and Nardo, Italy,
were investigated for fecal-contamination
indicators, bacterial pathogens, and
antibiotic resistance genes over the period
of one year. Real-time quantitative PCR
assays for Helicobacter pylori, Yersinia
enterocolitica, and Mycobacterium avium
subsp. paratuberculosis, human pathogens
with long-time survival capacity in water,
and for the resistance genes (ermB, mecA,
blaSHV-5, ampC, tetO, and vanA) were
adapted or developed for water samples
characterized by different impacts.
The resistance genes and pathogen
concentrations were determined at five
or six sampling points for each recharge
system. The three aquifer recharge
systems demonstrated different capacities
for removal of fecal contaminants and
antibiotic resistance genes.
A targeting species-specific PCR assay
was combined with a filter system to
collect phytoplankton cells and
spatial and temporal series as part of the
Mediterranean Sea EU project Strategy
(61). The application of PCR allowed a rapid
detection of several harmful dinoflagellate
species and genera, including Alexandrium
spp. Field samples were concentrated
on filter membranes, total DNA was
extracted from mixed phytoplankton
populations and PCR assays were carried
out with specific primers. Qualitative
PCR results were compared with light and
epifluorescence microscopic examinations.
Results indicated that this molecular
assay was able to detect harmful target
cells at concentrations undetectable by
microscopy. The application of this filter
PCR assay to seawater samples showed to
be a sensitive and rapid procedure for the
routine monitoring of coastal waters.
3.3 Ecological community studies and
The classification of the quality status
of a water body through the study of
the resident ecological community at
different tiers is currently widely diffused
in European monitoring programs also
thanks to the impulse received by WFD.
The effect of stressors on the ecological
community is evaluated both at structure
and function levels. Macrophytes, diatoms,
macrobenthos and fish are the mostly used
ecological quality indicators.
Among the other biological components
of the ecological community in both water
bodies and soils, special attention should
be paid to the microbiological community
for its functional role, ubiquitous presence
and possible direct impact on human
In fact, the study on the natural capacity
of bacterial communities to remove
xenobiotics (pesticides, pharmaceuticals,
biocides) in soil and water could have a very
important implication for human health
protection. The CNR-IRSA is currently
studying natural bacterial communities
from contaminated sites: specific bacteria
strains that, after repeated exposure
to xenobiotic, adapted themselves and
became able to remove pollutants through
metabolic and/or cometabolic processes,
were isolated and identified (62-67). This
“self-purification” ability can be used for
“recovery strategies” (i.e. bioremediation)
CNR Environment and Health Inter-departmental Project
of contaminated sites.
The knowledge of the microbiological
quality of coastal waters and marine
organisms (fish, crustaceans and molluscs)
(68-70), that is fundamental to assess the
sanitary and ecological risk in a coastal
zone, is a central research issue for the CNR
Institute of Coastal Marine Environment
A recent work by Caruso et al. (71-75) is
focused on the use of bacterial indicators
to assess the anthropogenic pressures over
coastal aquatic environments. Selected
bacterial species (Escherichia coli,
Enterococcus spp.) or related parameters
allow to track the occurrence and evolution
of bacterial pollution, and to prevent
human health risks caused by the use of
polluted waters.
Up-to-date standard procedures for
bacterial pathogens determination and
identification are necessary, due to the
limitations of the usual culture methods
(long response times, low specificity). In
recent years, research efforts have been
devoted to the improvement of technical
equipment (automatic multiple samplers)
and methodologies for the assessment
of seawater microbiological quality
particularly addressing the detection
and enumeration of Escherichia coli or
Enterococcus spp. in seawater as faecal
pollution indicators. Rapid methods such
as the fluorescent antibody method and the
β-glucuronidase assay have been developed
and optimized to monitor bathing waters,
allowing the quantitative measurement of
target bacterial molecules and accurate
phenomena. Combined fluorescencestaining protocols have also been set up, in
order to detect bacteria which may present
a pathogenic potential. Data obtained by
these new analytical procedures encourage
the use of E. coli or related parameters
as successful tools for early warning
of seawater bacterial pollution and for
the screening of polluted coastal areas;
therefore they offer interesting perspectives
to prevent waterborne diseases.
coastal waters is usually estimated by
determination of faecal indicator bacteria.
However, bacterial species possibly
pathogen for humans could occur also
as microorganisms indigenous to coastal
marine environments (e.g. Vibrio spp.).
Since their concentration is related to
the temperature of coastal waters, and
unrelated to classical faecal indicators,
monitoring and control of this bacterial
group is needed to plan preventive
measures for human health protection.
(70,76-78) The Vibrio genus is widespread
in coastal waters and includes more than 63
species. The most well-known species is V.
cholerae, which causes cholera epidemics
worldwide. In addition to V. cholerae,
many other Vibrio spp. are recognized
as potentially human pathogens causing
3 major syndromes of clinical illness:
gastroenteritis, wound infections, and
septicemia. Epidemiologic data suggest
that the majority of these infections are
foodborne disease and associated with
raw or undercooked shellfish. In wild and
cultivate shellfish the bacterial density
may reach high concentration and potential
toxic effect for humans. For this bacterial
group the standard microbiological criteria
used to assess water quality have to be
The analysis of antibiotic resistance
patterns (ARPs) of faecal indicator
bacteria allows to detect the presence and
persistence in the environment of genes
linked to antibiotic resistance. Research
is in progress to characterize the ARPs
of enterococci, as they are emerging
pathogenic bacteria, and of E. coli for
Water and Soil Monitoring for the Protection of Environment and Human Health
their capacity of acquiring antibiotic
resistance and spreading their resistance to
other species such as Salmonella, Shigella,
Yersinia, Vibrio etc. (79-81).
Studies on E. coli and enterococci have
particular epidemiological and ecological
relevance because these micro-organisms
can occupy multiples niches, including
humans, mammals, birds, reptiles and
3.4 Advanced technologies and early
warning systems
Advanced technologies have been applied to the
monitoring of soil and water for the protection
of human and environment health.
Among those, interest is growing for the
application of innovative microscopy
techniques, such as scanning probe
microscopy (AFM) and scanning near-field
optical (SNOM) to the environmental field.
These techniques exploit the interaction of
a functional tip scanned over the surface of
a sample (solid or adhesive on the substrate)
to reconstruct the morphology of the
sample and, simultaneously, achieve superresolution maps of other properties of the
sample (i.e. local optical and fluorescence
properties, maps of surface friction and
nano-mechanical properties, magnetic
domains, etc.). These microscopes are
very useful in the study of nanostructured
materials, surfaces and interfaces (i.e.
surfaces), and analysis of nanoscale
biosystems that can be investigated
at the cellular level, subcellular and
macromolecular aggregates. In particular,
biosystems can be studied in physiological
conditions and can describe temporal,
evolution (morphometry). (82-88) In
addition to the development of innovative
tools, expertise has been gained over the
years in the study of intra-cells interactions
in culture and exposed to environmental
agents (drugs, electromagnetic fields,
heavy metals, UV etc.) or nanostructured
agents (nanoparticles or nanotubes) and in
the study of complex phenomena such as
cellular aging or apoptosis.
An important research field is the
integration of advanced technologies in
measuring systems that can be used for
on-line or in-situ monitoring. The final
goal is to make available some devices
that can act as an early warning system
of sudden alteration in the environmental
quality status.
In the CNR-IRSA, Liquid Chromatography
with Mass Spectrometric detection (LCMS) has been integrated with an automatic
sampling and preparation station into
an on-line monitoring station for the
determination of polar organic substances
in drinking water (89). This system can
be used in potable water treatment plants
for the control of influent and effluent. In
this way, a control on the incoming water
quality and efficiency of the treatment
procedure should be achieved. The
operating limitations of this station are
linked to the total cost of the equipment, that
is still too high for a massive deployment,
the reduced frequency of sampling and the
need for highly qualified professionals for
the frequent maintenance required.
Spectroscopic techniques are suitable to
be integrated in small portable or fixed
station devices for in-situ monitoring by
optical fiber detection.
The concentration of several pollutants,
usually present in industrial waste
waters, could be predicted by the neural
network data processing of absorption
and fluorescence measurements in the
visible spectral range. Proper network
tuning provides quantitative analysis of
many pollutants with sub-ppm resolution.
CNR Environment and Health Inter-departmental Project
Compact optical fiber instrumentation for
absorption spectroscopy and an innovative
flowcell for fluorescence measurements
enable cost-effective, in situ, nonstop
monitoring of waste waters (90).
In the framework of the development of new
methods for measuring and monitoring soil
pollution, the use of magnetic susceptibility
as a proxy variable for monitoring heavy
metals in soils has been explored (91).
Magnetic measurements are carried out
by using a magnetic susceptibility meter
with two different probes for in situ field
surveys. The relationships between heavy
metal levels and magnetic susceptibility
values of soil samples were assessed.
Results suggest that a careful check of the
experimental procedure plays a crucial
role for using magnetic susceptibility
measurements in situ monitoring of heavy
some tools capable of providing, over a
relatively short time, integrated responses
regarding the levels of contamination and
the ecological consequences on different
compartments of the concerned ecosystems.
The biosensor “tool”, that responds
precisely to this necessity, consists of a
biological active element - from an isolated
enzyme to a whole organism - immobilized
on a transducer system (sensor) for the
selective and reversible determination of
the presence and/or the concentration of
certain chemical molecules in a sample.
In fact selectivity and sensitivity, together
with the possibility to have a portable tool
are the main advantages of biosensors.
A compact and portable sensing device
that combines the production and detection
of hydrogen peroxide in a single flow assay
has been proposed for herbicide detection
in water (92). The principle on which the
biosensor is based is that herbicides, under
illumination, can inhibit the photosystem
II electron transfer. Photosynthetic
membranes isolated from higher plants
and photosynthetic micro-organisms,
immobilized and stabilized, can serve as
a biorecognition element for a biosensor.
The inhibition of photosystem II causes
a reduced photoinduced production of
hydrogen peroxide, which can be measured
by a chemiluminescence reaction with
luminol and the enzyme horseradish
Systems that use biological responses to
detect environmental quality changes in
continuous (on-line monitoring) in a simple,
quick and economical way can be used as
Biological Early Warning System. In this
field, the possible use of an electrochemical
growth signal transduction of a natural
biofilm (microecosystem) on a suitable
metallic substrate, was recently investigated
with the aim of revealing, in real time, any
alteration of the normal development of
the microbial community induced by the
presence of toxic substances in the aquatic
environment (93-95). The prototype of
the innovative patented biosensor shows
that the biological electrochemical signal
is significantly inhibited in the short term
(minutes-hours) by known concentrations
of a series of reference toxics.
Among the most advanced early warning
systems, based on in vivo response of
organisms to toxic agents, a Swimming
Behavior Recorder System - able to
measure the swimming behavior of
marine invertebrates larvae exposed to
toxic substances and/or environmental
matrix under controlled conditions - has
been presented (96-99). The methodology
for the detection of alteration in swimming
uses a prototype system to analyze
video graphics, capable of automatically
recording the aquatic organisms swimming
speed, and providing, on the basis of the
alteration of this parameter compared with
Water and Soil Monitoring for the Protection of Environment and Human Health
a control, two toxicological endpoints:
immobilization (acute) and alteration of
swimming speed (sub-lethal).
3.5 Auxiliary techniques for monitoring
Geophysical techniques have been used
for several years to measure hydraulic
parameters in the monitoring and
control of groundwater contamination.
The most common techniques are the
electromagnetic ones that are more
sensitive to any changes in physical
parameters (i.e. electrical conductivity) of
soils and sub-soils caused by the presence
of a particular contaminant in the water
tablet or soil porosity. Therefore, the use
of geophysics monitoring systems should
be able to follow over time and space
areas the evolution of a particular case of
pollution, such as, for example, a leakage
from a dumping site. The study and
application of these non-invasive and lowcost technologies, integrated with the more
traditional ones (sampling and diagnostic
studies), will lead to a knowledge of the
land and the environment, in order to
provide a better safeguard level and to plan
remediation procedures.
In the Hydrogeosite laboratory of the
CNR Institute of Methodologies for
Environmental Analysis (CNR-IMAA) a
simulation plant has been built to perform
hydrogeophysical experiments for the
integrated study of contaminated soils and
subsoils with the aim of creating a standard
methodology for practical intervention
The activity carried out by the CNR
Research Institute for Geo-hydrological
Protection (CNR-IRPI) in the field
of erosion and hydro-meteorological
monitoring is of fundamental importance
for the protection of the water bodies and
their basin.
The literature review highlights that the
monitoring focus is still mainly addressed
to some classes of well studied molecules,
such as trace metals and persistent organic
pollutants, which represent a typology of
pollution that emerged several decades
ago, but for which there is still concern
because of their persistence in soil and
water sediments and their capacity to
accumulate in the trophic chain.
However there are many relatively new
classes of pollutants, which we daily
deal with, but are not still regulated by
legislation. For these classes, fundamental
research and appropriate monitoring plans
are needed in order to understand the
environmental distribution and fate, which
are necessary to assess the effective risks
for ecosystem and humans. CNR Institutes
are carrying out research on many of these
emerging environmental issues and, among
them, two classes can be chosen as case
studies, i.e. the engineered nanoparticles
and the perfluorinated compounds. In the
following sections a short critical review
of the knowledge gaps will be presented in
order to list future research needs for these
two emerging classes of compounds.
4.1 Engineered nanoparticles
Anthropogenic nanoparticles (engineered
nanoparticles; ENPs) are used in
nanotechnology to create products used
in various fields such as agriculture,
electronics, biomedicine, manufacturing,
pharmaceuticals cosmetics industry. The
use of nanomaterials containing ENPs is
expected to continuously increase in the
near future. The nanometer size range
(0-100 nm) means that nanomaterials
exhibit properties and functions other than
those owned by the same materials with
larger diameter and these properties are
CNR Environment and Health Inter-departmental Project
Figure 2: ENPs’ environmental fate (figure adapted from National Institute for Resources
and Environment, Japan
attributable to the increase of the surface the methods for their determination and
area ratio and number per unit mass of study in contaminated environments are
ENPs, which lead to increased chemical not yet standardized.
reactivity, greater strength and electrical The ENPs’ environmental fate is extremely
conductivity and, potentially, a more complex and the processes regulating ENPs’
distribution in the different compartment are
pronounced biological activity.
The extensive use of anthropogenic still not exhaustively investigated (Fig. 2).
There are physico-chemical processes that can
nanomaterials in large consumption
affect their potential environmental toxicity
products means that their transport, use (solubility, aggregation, absorption, interaction
and discharge is a potential new source of with other toxic substances). The study of
pollution in domestic sewage and industrial ENPs physico-chemical behavior influenced
discharges, resulting in a diffused pollution by abiotic factors in different matrices is
of surface waters and transitional/coastal fundamental to simulate scenarios in laboratory
areas by ENPs. ENPs can be transferred to experiments.
humans through food and several studies According to Moore, some open issues can be
(103-120) demonstrate their potential risk suggested for future research projects(121):
• Which is the hydrodynamic behaviour of
for human health.
Today there are still many uncertainties
• Do they behave like larger natural
about the environmental fate and toxicity
of ENPs in aquatic environments because • How do they associate with larger sediment
Water and Soil Monitoring for the Protection of Environment and Human Health
Figure 3: Different levels of biological organization in a laboratory mesocosm (by Dr.
Giovanni Pavanello, ISMAR).
and natural colloidal particulates?
international research institutions.
• Do they bind lipophilic organic and metal For the evaluation model, different classes
pollutants, and what are the routes of of anthropogenic ENPs representative
nanoparticle uptake into biota?
of potential future scenarios of use and
• Do ENPs-linked chemical pollutants show
impact in the aquatic environment should
enhanced toxicity?
be chosen. Their interaction with various
• Are the particle size and surface properties
significant factors in determining toxicity compartments of aquatic ecosystems
(water and sediment), in varying chemical
in aquatic organisms?
• What is the implication of ENPs exposure and physical conditions (pH, temperature,
for organisms health and ecosystem ion
concentration) should be determined.
• Can modelled fluxes and predicted impacts For each ENPs’ selected class a
of ENPs help to provide an explanatory standardized protocol for measurement
framework for their environmental must be developed for each different
behaviour and possible impacts?
environmental matricx (water, sediment
All these gaps could be filled by planning and biota), including all the possible
an appropriate laboratory experimental chemical and physical interactions between
model which should involve different ENPs and the different matrices. One
expertises available in the CNR Institutes, approach could be a careful and critical
with the cooperation of other national and
CNR Environment and Health Inter-departmental Project
analysis/testing of ENPs’ “detection”
methods already used in biomedical
field (Environmental Scanning Electron
Gun-Environmental Scanning Electron
Transmission Electron Microscope-STEM,
Transmission Electron Microscope-TEM,
Scanning Electron Microscope-SEM,
Atomic Force Microscope-AFM, Scanning
Laser Scanning Microscope-CLSM and
MultiPhoton Microscope-MPM) in order
to adapt them to the specific peculiarities
of the environmental matrices.
A crucial issue for a reliable risk
assessment on ecosystems is the selection
and standardization of ecotoxicological
bioassays at different levels of biological
organization (molecules, cells, organisms),
which must be representative of different
aquatic environments and ENPs categories
selected as references standards. The
selection of model species should be large
enough to ensure a good representation
of the different compartments of the most
representative reference ecosystems.
Although it is reasonable and practical
to predict the potential environmental
risk on the basis of bioassays with single
species models and/or battery of them,
it is equally important to investigate
how these supramolecular entities
interact at ecosystem level. For this
purpose, investigation protocols and
exposure methods using realistic ENPs
concentrations for various environments
should be developed using laboratory micro
and mesocosms containing sediment,
microorganisms, algae, invertebrates
and vertebrates, with different levels of
ecosystem complexity. (Fig. 3).
The mesocosm study can also supply
information on the accumulation and
eventual biomagnification along the trophic
chain. These data, together with human
dietary and environmental exposure
studies, are needed to get a reliable risk
assessment for human health.
4.2 Perfluorinated compounds in the water
Perfluorinated alkyl substances (PFASs)
are fluorinated man-made chemicals with
unique molecular properties, chemical and
thermal stability, water- and fat-repellent
properties that make them and their
derivatives useful in different industrial
and household applications. They are
widely employed for impregnation of
textiles and paper, for cleaning aids, for fire
fighting foams, for metal surface treatment
and in the production of fluoropolymers.
Among PFASs, perfluorooctanoic (PFOA)
and perfluorooctanesulfonic (PFOS) acids
are two persistent and bioaccumulative
end-stage metabolites of particular concern
for human health and the environment
due to their potential toxicological effects
on animal and also human endocrine
system; to date, there is no evidence of
biodegradation of these chemicals.
Recent concerns with the toxic effects
of PFOS and PFOA began in the early
2000s when the 3M Company, U.S.A, the
major perfluorochemical manufacturer,
decided to phase out the production of
PFOS related products. However, there are
still a number of industries, such as the
semiconductor etc, that still use PFOS in
their production (122) and manufacturers
that use and produce PFOA and its related
products for consumer usage. Evidence is
accumulating regarding their persistence in
the environment, potential for long-range
transport, tendency to bioaccumulate, and
potential toxicological effects (123-125).
Due to their physico-chemical properties,
these chemicals leaked into the water, they
accumulate in surface waters, and water
Water and Soil Monitoring for the Protection of Environment and Human Health
is the major reservoir of perfluorinated
compounds (PFCs) in the environment,
as well as the most important medium
for their transport (126,127). Unlike other
classical persistent organic pollutants
(POPs), the anphiphilic PFCs have not
been shown to accumulate preferentially
in adipose tissue. They rather bind to
blood proteins and accumulate in the liver
of exposed organisms (128). Recent studies
have indicated that PFOS can biomagnify
in higher trophic organisms through the
aquatic food chain (129,130).
PFOA is mainly used as adjuvant in
the production of fluoropolymers such
as polifluorotetraethylene and similar
products used in clothing production,
cosmetics and for non-stick cookware
coatings. It was estimated that aqueous and
gaseous emissions of PFOA or their salts
originated in this production represent the
majority (about 80%) of environmental
release (126). It is also used (but in small
amount) in industrial and consumer
products (varnishes, inks, paper, floor
polishes, cleaning formulation etc.) which
represent some direct human exposure
routes and diffusive environmental
sources not normally considered. PFOA
may also form as a degradation product
of fluorotelomer alcohols found in a wide
range of household and consumer products
like hair shampoo, rug cleaners, and food
paper products. These are volatile and
can be carried for long distances by air
currents (131).
In a European study on the major EU rivers
carried out by the Perforce consortium
(PERFORCE project), river Po watershed
was identified as the dominant source of
PFOA in Europe. River Po accounted for
two thirds of the total PFOA discharge
of the rivers studied (127). The PFOA
concentration (200 ng/L) in the water of
the Po river collected at the basin closure
(Pontelagoscuro near Ferrara) was from 10
to 200 times higher than those measured
in the other European rivers. This result
suggested is the presence of an industrial
source of PFOA in the Po watershed.
Recent survey of perfluorocarboxylic and
perfluorosulfonic acids concentrations
in the Po watershed (33,132) confirmed
the data of the PERFORCE project, and
measured a PFOA concentration of 60174 ng/l at Pontelagoscuro (FE). All
tributaries, except river Tanaro, showed
the PFOA levels typical of a diffuse
pollution; on the contrary, the highest
PFOA concentrations were determined in
river Tanaro (1270 ng/L) and in river Po,
downstream the confluence of the Tanaro
river (60-337 ng/L), suggesting the presence
of a point source for this compound that can
be reasonably attributed to an industrial
Teflon production site in the Alessandria
In the study carried out at the CNR-IRSA
(33) an assessment of the contamination
by perfluoroalkyl acids (PFAAs) in
surface, urban and industrialized waste
waters and drinking waters in the river
Po basin was carried out. An HPLC
method with ion trap mass spectrometry
detection was developed for the analysis
of perfluorinated carboxylates (from C5 to
C10) and perfluorinated sulfonates (C4 and
C8) with a LOQ of 2 ng/L. Water sample
extraction was performed on weak anion
exchange (WAX) cartridges.
In this work three kinds of waste waters,
from textile industry discharge, urban
wastewater treatment plant (WWTP)
and mix urban-industrial WWTP, were
analyzed because the industrial activity
is considered the main source of PFOA
and other homologues. PFOS was found
at lower concentrations than PFOA, both
in industrial and urban waste waters, This
is probably due to the phase out of the
CNR Environment and Health Inter-departmental Project
Figure 4. Perfluorinated concentration in
tap waters. LOQ = Limit of quantification
(2 ng/L); NJDEP = New Jersey Department
of Environmental Protection guidance
level (40 ng/L)
PFOS and PFOS-related compounds in the
electrochemical production since 2002.
Three sources of tap waters (Lecco produced from Lake of Lecco water;
Milano - produced from ground water;
Ferrara - partially produced from the
Po river) were also analyzed. The only
in drinking water above the limit of
quantification, was PFOA in the tap water
of Ferrara which produces drinking water
mixing ground and surface waters from
river Po (Fig. 4).
To date, there are no international water
guidelines for PFOA in drinking water
but some single countries set up their own
water guidelines. The strictest standard,
(40 ng/L), issued by New Jersey in the
USA, was not overcome by Ferrara tap
waters. Nevertheless it is necessary to
underline that this sampling campaign
on drinking waters does not have any
statistical representativeness, but it is only
a baseline survey which needs further in
depth enquiry.
Preliminary results highlighted the
presence, in the Po river watershed, of point
and diffused sources of perfluorinated
compounds. PFAAs were measured
in drinking waters produced from
contaminated surface waters, revealing
the risk for human consumption.
These preliminary campaigns, carried
out in different periods and hydrological
regimes, gave only a first look at the
distribution of perfluorinated compounds
in the Po watershed but no data have
been collected for the rest of Northern
Italy whose waters discharge in the High
Adriatic Sea. The available data do not
allow to estimate the risk of contamination
for the aquatic species present in the
transitional and coastal areas.
Furthermore the presence of intensive
aquaculture activities in these areas
could result in a source of exposure also
for humans which, at the present state of
knowledge, can not be envisaged.
Therefore, the overall goal of the IRSA
research group will be to assess the
environmental and health risks posed by
the PFCs contamination in the river Po
basin and in other river basins and coastal
areas of the Northern Adriatic sea. In order
to reach these objectives, the following
detailed actions should be carried out:
• a comprehensive survey of the river Po
basin , the coastal areas and lagoons of
the High Adriatic Sea in order to identify
critical areas and hot spots for PFC impact
o Monitoring the river Po basin with
downstream sampling of the main
o Evaluation of PFCs distribution in surface
and groundwater around fluorochemical
industrial sites in Northern Italy;
o Monitoring of mollusc aquaculture areas
in hot spot areas such as the Po Delta and
Northern Adriatic Sea coast
o Monitoring of raw and treated drinking
water drawn from the Po river or
groundwater impacted by industrial
• intensive monitoring of the critical areas
in different hydrological regimes for the
assessment of contamination and transport
Water and Soil Monitoring for the Protection of Environment and Human Health
• determination of PFCs levels in edible
marine and lagoon bivalves, farmed or
caught in the critical areas, in order to
o PFCs
sites, using two edible species with
different geographical distribution (site
o PFCs accumulation in two different
bivalves, addressing the possible existence
of different patterns of exposure (clams
in the sediment/mussels in the water
column) and of bioaccumulation (species
• Estimation of PFCs risk exposure for
humans as a result of the consumption
of contaminated mollusks and drinking
The term “emerging pollutants” means
a continuously updated list of the most
diverse molecules produced, used and
eventually diffused in the environment.
Such list includes synthetic surfactants,
endocrine disruptors, pharmaceutical
substances, perfluorinated compounds,
industrial additives, new generation
pesticides, etc, all substances which are not
included in the lists of regulated priority
pollutants and in ordinary monitoring
plans. Considering also that the toxicity
and the effects of certain new generation
compounds are often not known or fully
understood, it is evident how crucial it is
to develop monitoring systems suitable to
detect the possible responses to unknown
toxic agents. In fact it is impossible to
measure all the compounds present in
a certain environmental compartment
and we need to turn over our traditional
approach in monitoring: first we measure
the effects and then we look for the
molecular agents which should be the
cause of the effect itself. This revolution in
the monitoring approach will be probably
the most outstanding innovation in the
next future.
At present, the procedures identified by
the words “Toxicity Identification and
Evaluation” (TIE) or “Effect Directed
Analysis” (EDA) represent the state of the
art in this field and for that reason we will
present a short review on this topic in this
last section.
The first identification process of
“guilty” substances and its verification
of the observed effects caused by these
compounds dates back to 1983 (133).
Subsequently taken over by the United
States Environmental Protection Agency
(US EPA), this process took the name of
Toxicity Identification Evaluation (TIE).
The classic TIE is divided into 3 phases.
During the first phase (Fig. 5a), groups
of active compounds are sequentially
removed from the water sample with
chemical treatment, as long as the toxic
effect of the solution on the biological
model system does not disappear. In the
second phase substances are tentatively
identified by the treatment that decreases
the toxic response of the sample and the
classes of chemicals identified are related
to the biological responses. Finally the
activity of each identified substance or
substances mixture is confirmed using the
same biological model.
The Effect Directed Analysis (EDA)
procedure, which is the subject of EU
research projects in the framework of WFD
implementation, is the conceptual and
technical evolution of the TIE approach.
Like TIE, EDA is based on the use of the
response of a biological reference system
in combination with sample fractionation
and the chemical analysis of individual
fractions for identification of hazardous
compounds in complex mixtures present in
the environment. Figure 5b schematically
CNR Environment and Health Inter-departmental Project
Figure 5: a-An example of TIE’s first phase. b-scheme of EDA process.
describes the process which starts from the
observation of a “macroscopic” effect on
the biotic component in the investigation
site. Through the use of a chromatographic
separation systems, the mixture is divided
into its individual components. Each
fraction is tested for activity on in-vitro
model systems, the nature of the compound
is defined through analytical detection
and recognition systems to determine the
substance responsible for the effect.
Sample fractionation into individual
classes of compounds is achieved using
chromatographic columns with different
stationary phases based on the principle that
each compound has a specific affinity with
the stationary phase column at a certain
mobile phase composition. Therefore
different types of serially connected
chromatographic columns allow a sample
basic components separation efficiency
greater than that obtained using a single
column, as in simpler TIE procedures
(134,135 ).
The identification and confirmation
of compounds in each fraction is
achieved by using high-resolution mass
spectrometry techniques, hyphenated
with chromatographic systems; liquid
chromatography (HPLC) is used for the
separation and identification of polar
compounds and metabolites which are
more difficult to analyze but are much more
biologically active than classic persistent
organic pollutants.
In addition to a more rigorous procedure
and a greater degree of complexity and
precision in the sample fractionation
compared to TIE, EDA introduces the
use of in-vitro bioassays that allow a high
number of spatial and temporal replicates
and provide the capacity to run the tests in
batteries, so getting a complete screening
of the toxic activities of the sample fraction.
As examples of in vitro tests used in EDA
approach we can mention Yeast estrogen
screen (YES), Yeast androgen screen
(YAS), for the evaluation of endocrine
disruption activity (136), bioluminescence
tests such as CALUX, Microtox and
Mutatox, assays on fish hepatocytes for
physiological and accumulation studies:
these are all sensitive, specific and rapid
tests which allow to provide a full view on
the biological activity of the compounds
under investigation.
In parallel to those “classic” in-vitro
tests, in recent years, the development of
the “omics” disciplines helped to expand
knowledge about the effects of many
Water and Soil Monitoring for the Protection of Environment and Human Health
toxic compounds: toxicogenomics and
toxicoproteomics study the relationships
between genome (or protein) structure
and activity and the biological effects of
exogenous agents. This means that if a
compound shows, for example, a mutagenic
effect, the magnitude of the effect can be
determined by the gene expression analysis
of an exposed individual.
As an example of the effectiveness of
the integrated monitoring and analysis
approach, we can mention the study
of Keiter et al. (137) who adopted an
integrated TRIAD and EDA method to
identify the activity of so-called dioxinlike compounds (i.e. molecules whose
steric and electronic structure is similar to
those structural characteristics of dioxins
which exert effects on biological systems)
as the cause of a specific adverse effect on
river organisms.
The EDA procedure is rather complex
and time consuming and, of course, it is
not suitable to meet the demand of fast
and reliable techniques for early warning
systems. Nevertheless, if integrated
into routine monitoring programs as a
complementary technique for investigative
monitoring, it will be absolutely necessary
in those situations where the toxic or other
biological effect cannot be attributed to
a known agent or source. Alternatively it
could be used as a screening method to
assess the risk of a particular pollution
source that is crucial for an effective
management of the territory with the
aim of human health and environmental
With the contribution of WG2 participants: A.
Barra Caracciolo (IRSA), E. Belluso (IGG)
G. Caruso (IAMC), L. Da Ros (ISMAR),
M. Faimali (ISMAR), M. Girasole (ISM),
R. La Ferla (IAMC), M. Mancuso (IAMC),
G. Mascolo (IRSA), V. Matranga (IBIM),
L.S. Monticelli (IAMC), E. Morelli (IBF), P.
Gualtieri (IBF), M. Pennisi (IGG), M. Polemio
(IRPI), E. Rizzo (IMAA), E. Sacchi (IGG), R.
Zaccone (IAMC).
With the contribution of: A. Baldi (Fondazione
Italiana Endometriosi); E. Fommei (Pisa
University and CNR-IFC), G. Iervasi, A.
Pierini (CNR-IFC); S. Maffei, C. Vassalle
(Fondazione “G. Monasterio” CNR-Regione
Toscana, Pisa); C. Roscioli, S.Valsecchi, L.
Viganò, D. Vignati (CNR-IRSA, Brugherio).
Keywords: water and soil monitoring,
biomarker, bioassay, early warning system,
emerging substances.
SETAC Society of Environmental
toxicology and chemistry. Technical
issue paper: Ecological risk assessment.
Pensacola (FL): SETAC 1997: 4p.
Chapman PM. The sediment quality
Triad approach to determining pollutioninduced degradation. Sci Total Environ
Long ER, Chapman PM. A Sediment
Quality Triad: Measures of sediment
contamination, toxicity and infaunal
community composition in Puget Sound.
Mar Poll Bull 1985;16:405–415.
Spurgeon DJ, Ricketts H, Svendsen C,
Morgan AJ, Kille P. Hierarchical responses
of soil invertebrates (earthworms) to
toxic metal stress. Environ Sci Technol
Dagnino A, Allen JI, Moore MN, Broeg
K, Canesi L, Viarengo A. Development
of an expert system for the integration
of biomarker responses in mussels into
an animal health index. Biomarkers
Viarengo A, Gastaldi L, Dagnino A,
Capri F, Torrielli S, Pons G. An expert
system assessing pollutant-induced stress
syndrome in the earthworm Eisenia
andrei. Proceedings of the Society of
Environmental Toxicology and Chemistry
(SETAC) 17th Annual Meeting; 2007
May 20–24; Porto, Portugal. Pensacola
CNR Environment and Health Inter-departmental Project
(FL): SETAC. 184 p.
Spurgeon DJ, Hopkin SP, Jones DT.
Effects of cadmium, copper, lead and zinc
on growth, reproduction and survival of
the earthworm Eisenia fetida (Savigny):
Assessing the environmental impact
of point-source metal contamination in
terrestrial ecosystem. Environ Pollut
Dickerson RL, Hooper MJ, Gard NW,
Cobb GP, Kendall RJ. Toxicological
foundations of ecological risk assessment:
Biomarker development and interpretation
based on laboratory and wildlife species.
Environ Health Perspect 1994;102:65–69.
Ehlers LJ, Luthy RG. Contaminant
bioavailability in soil and sediment.
Environ Sci Technol 2003; 37:295A–
Semple KT, Doick KJ, Jones KC,
Burauel P, Craven A, Harms H. Defining
of contaminated soil and sediment
is complicated. Environ Sci Technol
Semenzin E, Temminghoff EJM,
Marcomini A. Improving ecological risk
assessment by including bioavailability
into species sensitivity distributions: An
example for plants exposed to nickel in
soil. Environ Pollut 2007;148:642–647.
Crumbling DM, Lynch K, Howe R,
Groenjes C, Shockley J, Keith L et al.
Managing uncertainty in environmental
McCarthy JF, Shugart LR. Biomarkers
of environmental contamination. Chelsea
Mich.: Lewis Publisher;1990.
Hagger JA, Jones MB, Leonard DRP,
Owen R, Galloway TS. Biomarkers and
integrated environmental risk assessment:
are more questions than answers? Integr
Environ Assess Manag 2006;2:312–329.
Dagnino A, Sforzini S, Dondero F,
Fenoglio S, Bona E, Jensen J et al. A
‘‘Weight-of-Evidence’’ Approach for the
Integration of Environmental ‘‘Triad’’
Data to Assess Ecological Risk and
Biological Vulnerability. Integrated
Management 2008;4(3):314-326.
Bretzel F, Calderisi M. Metal
contamination in urban soils of coastal
Tuscany (Italy) Environmental Monitoring
And Assessment 2006;118(1-3): 319-335.
Scerbo R, Ristori T, Stefanini B, De
Ranieri S, Barghigiani C. Mercury
assessment and evaluation of its
impact on fish in the Cecina river basin
(Tuscany, Italy). Environmental Pollution
Bianchini G, Pennisi M, Cioni R, Muti A,
Cerbai N, Kloppmann W. Hydrochemistry
of the high-boron groundwaters of
the Cornia aquifer (Tuscany, Italy).
Geothermics 2005;34:297-319.
Gonfiantini R and Pennisi M. The
behaviour of boron isotopes in natural
waters and in water-rock interaction. J
Geochem. Exploration 2006;88:114-117.
Pennisi M, Gonfiantini R, Grassi S,
Squarci P. The utilization of boron and
strontium isotopes for the assessment
of boron contamination of the Cecina
River alluvial aquifer (central-western
Tuscany, Italy). Applied Geochemistry
Pennisi M, Bianchini G, Muti A,
Klopmann W. Behaviour of boron and
strontium isotopes in groundwateraquifer interactions in the Cornia Plain
(Tuscany, Italy). Applied Geochemistry
Facchinelli A, Magnoni M, Perrone U,
Sacchi E. Post-depositional processes in
lake sediments traced by heavy metals
and radionuclides: a case study from Lake
Sirio (Ivrea, Northern Italy). Materials
and Geoenvironment 2005;52:31-33.
Sacchi E, Brenna S, Fornelli I, Genot S,
Sale VM, Azzolina L et al. Analisi del
contenuto in rame ed altri metalli nei suoli
agricoli lombardi.Quaderni della Ricerca
2007:61,Regione Lombardia,111.
Scarciglia F, Sacchi E, Angelone M,
Apollaro C, Armiento G, Barca D et
al. Caratteri geochimici, isotopici e
mineralogici dei suoli di Muravera. In:
Ottonello G. (Ed.) Geochemical Baselines
Water and Soil Monitoring for the Protection of Environment and Human Health
of Italy, Pisa, Pacini Editore 2007;87-147.
25. Sacchi E, Brenna S, Fornelli Genot S,
Setti M, Sale VM et al. A regional survey
on heavy metals content in cultivated
soils from Lombardy (Italy): results from
the RAMET project. Int. Symp. “Consoil
2008”, Milan (Italy), 2008 3-6 June 2008,
26. Cardellicchio N, Buccolieri A, Di Leo
A, Giandomenico S, Spada L. Levels of
metals in reared mussels from Taranto
Gulf (Ionian Sea, Southern Italy). Food
Chemistry 2008;107,(2):890-896.
27. Belluso E, Fornero E, Cairo S, Albertazzi
G, Rinaudo C. The application of microRaman spectroscopy to distinguish
carlosturanite from serpentine-group
28. Cardile V, Lombardo L, Belluso E,
Panico A, Capella S, Balazy M. Toxicity
and carcinogenicity mechanisms of
fibrous antigorite. International Journal
of Environmental Research and Public
Health 2007;4:1-9.
29. Cardile V, Lombardo L, Belluso E,
Panico AM, Renis M, Gianfagna A et al.
Fluoro-edenite Fibers Induce Expression
of Hsp70 and Inflammatory Response.
International Journal of Environmental
Research and Public Health 2007;20:195202.
30. Fornero E, Belluso E, Capella S, Bellis D.
Environmental exposure to asbestos and
other inorganic fibres using animal lung
model. Science of the Total Environment
2009; 407(3):1010-1018.
31. Detomaso A, Mascolo G, Lopez
A. Characterization of carbofuran
photodegradation by-products by liquid
time-of-flight/mass spectrometry. Rapid
Communication in Mass Spectrometry
32. Cavalli S, Polesello S, Saccani G.
Determination of Acrylamide in Drinking
Water by Large-Volume Direct Injection
and ICE-MS detection. Journal of
Chromatography A 2004;1039:155-159.
33. Valsecchi S, Polesello S. The search of
sources of perfluoroalkyl acids (PFAAs)
in Northern Italian waters, 19th Annual
Meeting of Setac Europe “Protecting
ecosystem health: facing the challenge
of a globally changing environment”.
Gőteborg, Sweden. 2009 May 31- June4.
Marin MG, Boscolo R, Cella A,
Degetto S, Da Ros L. Field validation
of autometallographical black silver
deposit (BSD) extent in three bivalve
species from the lagoon of Venice,
Italy (Mytilus galloprovincialis, Tapes
philippinarum, Scapharca inequivalvis)
for metal bioavailability assessment. Sci
Tot Environ 2006;371:156-167.
Da Ros L, Moschino V, Guerzoni S,
Halldòrsson HP. Lysosomal responses
and metallothionein induction in the blue
mussel Mytilus edulis from the south-west
coast of Iceland. Environ International
Neston N, Romano S, Moschino V, Mauri
M, Da Ros L. Chemical analysis and
biomarkers in mussels and fish as tools
for evaluating presence and effects of
microorganic pollutants and trace metals
in the lagoon of Venice, Italy. Mar Poll
Bull 2007;55: 469-484.
Angelini C, Amaroli A, Falugi C, Di Bella
G, Matranga V. Acetylcholinesterase
activity is affected by stress conditions
in Paracentrotus lividus coelomocytes.
Marine Biology 2003;143(4):623-628.
Russo R, Bonaventura R, Zito F, Schroder
HC, Muller I, Muller WEG et al. Stress to
cadmium monitored by metallothionein
gene induction in Paracentrotus lividus
embryos. Cell Stress & Chaperones
Bonaventura R, Poma V, Costa C,
Matranga V. UVB radiation prevents
skeleton growth and stimulates the
expression of stress markers in sea urchin
embryos. Biochem Biophys Res Commun
Bonaventura R, Poma V, Russo R, Zito F,
Matranga V. Effects of UV-B radiation on
the development and hsp 70 expression
in sea urchin cleavage embryos. Marine
Biology 2006;149:79-86.
CNR Environment and Health Inter-departmental Project
41. Matranga V, Pinsino A, Celi M, Natoli
A, Bonaventura R, Schröder HC et al.
Monitoring chemical and physical stress
using sea urchin immune cells. Prog Mol
Subcell Biol 2005;39:85-110.
42. Matranga V, Zito F, Costa C, Bonaventura
R, Giarrusso S, Celi F. Embryonic
development and skeletogenic gene
expression affected by X-rays in the
Mediterranean sea urchin Paracentrotus
lividus. Ecotoxicology [Epub ahead of
print]Online: DOI 10.1007/s10646-0090444-9. 2009 Nov 27.
43. Pinsino A, Matranga V, Trinchella F,
Roccheri MC. Sea urchin embryos as
an in vivo model for the assessment of
manganese toxicity: developmental and
stress response effects. Ecotoxicology
[Epub ahead of print]Online: DOI 10.1007/
s10646-009-0432-0. 2009 Nov 1.
44. Schröder HC, Di Bella G, Janipour N,
Bonaventura R, Russo R, Müller WE
et al. DNA damage and developmental
defects after exposure to UV and heavy
metals in sea urchin cells and embryos
compared to other invertebrates. Prog
Mol Subcell Biol. 2005;39:111-37.
45. Filosto S, Roccheri MC, Bonaventura
R, Matranga V. Environmentally
relevant cadmium concentrations affect
development and induce apoptosis of
Paracentrotus lividus larvae cultured
in vitro. Cell Biol Toxicol 2008;24: 603610.
46. Pinsino A, Della Torre C, Sammarini
V, Bonaventura R, Amato E, Matranga
V. Sea urchin coelomocytes as a novel
cellular biosensor of environmental stress:
a field study in the Tremiti Island Marine
Protected Area, Southern Adriatic Sea,
Italy. Cell Biol Toxicol 2008; 24: 541-552
47. Morelli E, Scarano G. Copper-induced
changes of non-protein thiols and
antioxidant enzymes in the marine
microalga Phaeodactylum tricornutum.
Plant Sci 2004;167:289–296.
48. Loreti V, Toncelli D, Morelli E, Scarano
G, Bettmer J. Biosynthesis of Cd-bound
Phytochelatins (PCs) by Phaeodactylum
tricornutum and their Speciation via
Size-Exclusion Chromatography (SEC)
and Ion-Pairing Chromatography (IPC)
coupled to ICP-MS. Anal Bioanal Chem
Morelli E, Mascherpa MC, Scarano
G. Biosynthesis of phytochelatins and
arsenic accumulation in the marine
microalga Phaeodactylum tricornutum in
response to arsenate exposure. BioMetals
Bramanti E, Toncelli D, Morelli E,
Lampugnani L, Zamboni R, Miller KE et
al. Determination and characterization of
phytochelatins by liquid chromatography
coupled with on line chemical vapour
generation and atomic fluorescence
spectrometric detection. J Chromatogr
Morelli E, Fantozzi L. Phytochelatins in
the Diatom Phaeodactylum tricornutum
Bohlin: An evaluation of their use as
biomarkers of metal exposure in marine
waters. Bull Environ Contam Toxicol
Morelli E, Marangi ML, Fantozzi L. A
phytochelatin-based bioassay in marine
diatoms useful for the assessment of
bioavailability of heavy metals released
by polluted sediments. Environ Int 2009
(in press).
Evangelista V, Barsanti L, Passatelli
Microspectroscopy of the Photosynthetic
Compartment of Algae.
Photobiol 2006;82:1039-1046.
Evangelista V, Evangelisti M, Barsanti
L, Frassanito AM, Passarelli V,
Gualtieri P. A polychromator - based
microspectrophotometer”. Int J Biol Sci
Rodriguez MC, Barsanti L, Passarelli V,
Evangelista V, Conforti V, Gualtieri P.
Effects of chromium on photosynthetic
and photoreceptive apparatus of the
Environmental Research 2007;105:234-9.
Barsanti L, Coltelli P, Evangelista V,
Passarelli V, Rassanito AM, Gualtieri
P. Low resolution characterization
of the 3D structure of the Euglena
Water and Soil Monitoring for the Protection of Environment and Human Health
gracilis photoreceptor. Biochemical and
Biophysical Research Communications
Pampanin DM, Marangon I, Volpato E,
Campesan G, Nasci C. Stress biomarkers
and alkali-labile phosphate level in
mussels (Mytilus galloprovincialis)
collected in the urban area of Venice
(Venice Lagoon, Italy). Environmental
Pollution 2005;136(1):103-107.
Pampanin DM, Volpato E, Marangon I,
Nasci C. Physiological measurements
from native and transplanted mussel
(Mytilus galloprovincialis) in the canals
of Venice. Survival in air and condition
index. Comparative Biochemistry and
Physiology A-Molecular & Integrative
Physiology 2005;140(1):41-52.
Albani A, Serandrei-Barbero R, Donnici
S. Foraminifera as ecological indicators
in the Lagoon of Venice, Italy. Ecological
Indicators 2007;7(2):239-253.
Bockelmann U, Dorries HH, AyusoGabella MN, de Marcay MS, Tandoi
V, Levantesi C et al. Quantitative PCR
Monitoring of Antibiotic Resistance
Genes and Bacterial Pathogens in
Three European Artificial Groundwater
Penna A, Fusco G, Bertozzini E,
Giacobbe MG, Vila M, Galluzzi L et al.
Monitoring of Alexandrium species in
the Mediterranean Sea using a combined
filter system-PCR assay detection method.
African Journal of Marine Science
Accinelli C, Barra Caracciolo A, Grenni
P, Giuliano G, Vicari A. Degradation of
the antiviral drug oseltamivir carboxylate
in surface water. In: Environmental Fate
and Ecological effects of pesticides. Del
Re AAM, Capri E, Fragoulis G, Trevisan
M Eds, La Goliardica Pavese 2007:401407.
Grenni P, Barra Caracciolo A, Saccà
ML, Falconi F, Ciccoli R, Ubaldi C et
al. Natural attenuation capability of an
agricultural soil to degrade terbuthylazine.
In: Environmental Fate and Ecological
effects of pesticides. Del Re AAM, Capri
E, Fragoulis G, Trevisan M Eds, La
Goliardica Pavese 2007:601-606.
Martín M, Gibello A, Martínez-Iñigo
MJ, Lobo MC, Nande M, Garbi C et
al. Proposal for a Natural Attenuation
Coefficient for simazine- contaminated
soils based on fluorescence in situ
hybridization. Chemosphere 2008;71:703710.
Singer A, Howar M, Johnson A, Accinelli
C, Bird S, Boucard T et al. Meeting
report: risk assessment of Tamiflu® use
under pandemic conditions - Report
from an interdisciplinary Workshop.
Environmental Health Perspectives
2008;116(11):1563 -1567.
Grenni P, Barra Caracciolo A, RodríguezCruz MS, Sánchez-Martín MJ. Changes
in the microbial activity in a soil amended
with oak and pine residues and treated
with linuron herbicide. Applied Soil
Ecology 2009;41:2-7.
Grenni P, Gibello A, Barra Caracciolo
A, Fajardo C, Nande M, Sacca ML et
al. A new fluorescent oligonucleotide
probe for in situ detection of s-triazinedegrading Rhodococcus wratislaviensis
in contaminated groundwater and soil
samples. Water Research 2009;accepted
Mancuso M, Avendaño-Herrera R,
Magariños B, Zaccone R, Toranzo AE.
Evaluation of different DNA-based
fingerprinting methods for typing
Photobacterium damselae subsp.piscicida.
Biol Res 2007;40(1):85-92.
Zaccone R, Mancuso M. First report on
antibodies response of Seriola dumerilii
(Risso 1810) challenged with Listonella
Immunology 2008;25(5):689-692.
Leonardi M, Azzaro F, Azzaro M,
Caruso G, Mancuso M, Monticelli LS et
al. Multidisciplinary study of the Cape
Peloro brackish area (Messina, Italy):
characterisation of trophic conditions,
microbial abundance and activities.
In: Marine Ecology: an evolutionary
perspective (S.Z.N.). M.C. Gambi and
CNR Environment and Health Inter-departmental Project
Levin eds; 2009. p.33-42.
71. Caruso G, Zappalà G, Crisafi E.
Monitoring bacterial pollution in coastal
waters: recent advances in technologies
and rapid methods”. 4th International
Conference on Marine Waste Water
Discharges and Marine Environment,
Antalya (Turkey), 2006 November 6-10,
Abstracts, 333-334.
72. Zappalà G, Caruso G, Azzaro F, Crisafi
E. Marine environment monitoring in
coastal Sicilian waters. In: Brebbia CA
editor. Water Pollution VIII: Modelling,
Monitoring and Management, Bologna,
2006 September 4-6;95:337-346. WIT
Press, Southampton (UK).
73. Caruso G, Crisafi E, Caruso R, Zappalà
G. Advances in marine bacterial
pollution monitoring. In: Environmental
Microbiology Research Trends, G. V.
Kurladze editor, Chapter 10, pp. 273-287,
NOVA Publishers: Hauppauge, NY. USA;
74. Caruso G, Monticelli LS, Caruso
R, Bergamasco A. Development of
a fluorescent antibody method for
the detection and enumeration of
Enterococcus faecium and its potential for
coastal aquatic environment monitoring.
Marine Pollution Bulletin 2008;56:318324.
75. Caruso G, Zappalà G, Maimone G,
Azzaro F, Raffa F, Caruso R. Assessment
of the abundance of actively respiring
cells and dead cells within the total
bacterioplankton of the Strait of
Messina waters. In: Brebbia CA editor,
Environmental Problems in Coastal
Regions VII, The New Forest (UK), 2008
May 19-21. Proceedings, pp.15-24. WIT
Press, Southampton (UK).
76. Zaccone R, Azzaro M, Azzaro F, Caruso
G, Giacobbe MG, Mancuso M et al. First
microbiological data from the lagoon area
of Cape Peloro (Messina). workshop 10
-131 Geoitalia 2007 Settembre 10-12.
77. Zaccone R, Azzaro M, Azzaro F, Caruso
G, Mancuso M, Monticelli LS et al.
Multidisciplinary study of Cape Peloro
brackish area (Messina, Italy) 43 EMBS
Azzorre 2008 September 8-12; P3.42.
78. Maimone G, Caruso G, La Ferla R,
Mancuso M, Zaccone R. Variabilità della
popolazione batterica in un ecosistema
salmastro della Sicilia. II Workshop
annuale VECTOR, 2009 February 25–
26; Roma.
79. Bergamasco A, Azzaro F, Caruso G,
Crisafi E, Decembrini F, Monticelli
LS et al. Tecniche e metodologie per la
valutazione dello stato ecologico delle
acque costiere e di transizione: risultati,
strategie e prospettive. 1° Forum Istituto
per l’Ambiente Marino Costiero, Giardini
Naxos (ME), 6-9 maggio 2007.
80. Caruso G, Zaccone R, Monticelli LS,
La Cono V, Crisafi E. Impatto antropico
su aree marine costiere: metodologie
innovative per il controllo igienicosanitario delle acque. 1° Forum Istituto
per l’Ambiente Marino Costiero, Giardini
Naxos (ME), 6-9 maggio 2007.
81. Caruso G, Monticelli LS, Caruso
R, Bergamasco A. Development of
a fluorescent antibody method for
the detection and enumeration of
Enterococcus faecium and its potential for
coastal aquatic environment monitoring.
Marine Pollution Bulletin 2008;56:318324.
82. Longo G, Girasole M, Cricenti A. A novel
tapping SNOM: Instrument description
and performances. Phys Stat Sol B
83. Moretti PF, Maras A, Palomba E, Girasole
M, Pompeo G, Longo G. Detection of
nanostructured metal in meteorites:
implications for the reddening of asteroids.
Astrophys J Lett 2005;634,L117-120.
84. Cattaruzza F, Cricenti A, Flamini
A, Girasole M, Longo G, Prosperi
T, et al. Controlled loading of
onto unoxydised crystalline silicon;
fluorescence-based determination of the
surface coverage and of the hybridisation
efficiency; parallel imaging of the
process by AFM. Nucleic Acid Research
85. Colonna S, Pompeo G, Girasole M,
Water and Soil Monitoring for the Protection of Environment and Human Health
Gazzoli D, Pettiti I, Valigi M. Thermallyinduced morphological transition in WOx
deposited on a ZrO2(100) substrate. Surf
Sci 2007;601:1389–1393.
Girasole M, Cricenti A, Generosi R,
Longo G, Pompeo G, Cotesta S et al.
Revealed by Atomic Force/Lateral Force
Microscopy After Doping of Human
Pancreatic Cells With Cd, Zn or Pb. Micr.
Res. Tech. 2007;70:912-917
Girasole M, Pompeo G, Cricenti A,
Congiu-Castellano A, Andreola F,
Serafino A, et al. Roughness of the
Plasma Membrane as an Independent
Morphological Parameter to Study RBCs:
a Quantitative Atomic Force Microscopy
Investigation. Biochim Biophys ActaBiomembranes 2007; 1768,1268–1276.
Longo G, Girasole M, Cricenti A.
Implementation of a bimorph-based
aperture tapping-SNOM with an incubator
to study the evolution of cultured living
cells. J Microscopy 2008;229:433-439.
Brumaru C, Polesello S, Valsecchi S. LCMS-Ion Trap determination of natural
and synthetic estrogens in drinking
water, 1st thematic workshop of the EU
project NORMAN – Chemical Analysis
of Emerging Pollutants; 2006 November
27-28; Maò, Menorca (Balearic island),
Spain; 2006. p. 101.
Kuzniz T, Halot D, Mignani AG, Ciaccheri
L, Kalli K, Tur M et al. Instrumentation for
the monitoring of toxic pollutants in water
resources by means of neural network
analysis of absorption and fluorescence
spectra. Sensor and Actuators B-Chemical
D’Emilio M, Chianese D, Coppola R,
Macchiato M, Ragosta M. Magnetic
susceptibility measurements as proxy
method to monitor soil pollution:
development of experimental protocols for
field surveys. Environmental Monitoring
and Assessment 2007;125(1-3):137-146.
Varsamis DG, Touloupakis E, Morlacchi
P, Ghanotaskis FD, Giardi MT, Cullen DC.
Development of a photosystem II-based
optical microfluidic sensor for herbicide
detection. Talanta 2008;77(1):42-47.
93. Garaventa F, Corrà C, Di Fino A,
Modugno S, Mollica A, Pavanello G, et
al. Automated systems to monitor marine
pollution, from eco-toxicological kit to online Biological Early Warning Systems: an
integrated approach. III Bilateral Seminar
Italy-Japan on: Physical and Chemical
Impacts on Marine Organisms - Seeking
sustainability and postgenomics; Nagoya
(Japan); 24-27 November 2008.
94. Faimali M, Chelossi E, Garaventa F,
Corrà C, Greco G, Mollica A. Evolution of
oxygen reduction current and biofilm on
stainless steels cathodically polarised in
natural aerated seawater. Electrochimica
Acta 2008;54(1)148-15.
95. Pavanello G, Pittore M, Mollica A,
Mollica A, Cappello M, Capparelli E et al.
Sviluppo di un biosensore elettrochimico
per la tossicità delle acque (beta), Biologia
Marina Mediterranea 2009;in press.
96. Faimali M, Garaventa F, Piazza V, Greco
G, Corrà C, D’Amico G. Mortality,
settlement inhibition and swimming speed
alteration of larvae of Balanus amphitrite
as acute, chronic and behavioural
end-point for laboratory toxicological
bioassays. Biologia Marina Mediterranea
97. Garaventa F, Corrà C, Di Fino AL,
Modugno S, Mollica A, Mollica A et al.
Automated systems to monitor marine
pollution, from eco-toxicological kit to online Biological Early Warning Systems: an
integrated approach. III Bilateral Seminar
Italy-Japan on: Physical and Chemical
Impacts on Marine Organisms - Seeking
sustainability and postgenomics – Nagoya
(Japan), 24-27 November 2008.
98. Meneghetti F, Garaventa F, Bon D, Di Fino
A, Gambardella C, Faimali M et al. Use
of marine invertebrates behavioural endpoints to evaluate the toxicity of coastal
sediment. International Expert Meeting
on The Impacts of Human Activities at
Sea, on The Coast and in Its Hinterland
on The Northern Adriatic’s Biodiversity –
Piran (SI), October 7th – 8th, 2008.
99. Gambardella C, Di Fino A, Garaventa
CNR Environment and Health Inter-departmental Project
F, Pittore M, Faimali M. Alterazione
del nuoto larvale di organismi marini
come end-point sub-letale in biosaggi
ecotossicologici. Biol Mar Medit 2009; in
100. Suski B, Rizzo E, Revil A. A sandbox
experiment of self-potential signals
associated with a pumping test. Vadose
Zone Journal 2005;3(4):1193-1199.
101. Bavusi M, Lapenna V, Rizzo E.
Electromagnetic methods to characterize
the Savoia di Lucania waste dump
(Southern Italy). Environmental Geology
102. Straface S, Fallico C, Troisi S, Rizzo E,
Revil A. Estimating of the transmissivities
of a real aquifer using a Self Potential
signals associated with a pumping test.
Ground Water 2007;45(4):420-428.
103. Oberdòrster G, Gelein RM, Ferin J, Weiss
B. Association of particulate air pollution
and acute mortality: involvement
of ultrafine particles? InhalToxicol.
105. Marconi A. Particelle fini, ultrafini e
nanoparticelle in ambiente di vita e di
lavoro: possibili effetti sanitari e misura
dell’esposizione inalatoria. G Ital Med
Lav Erg 2006;28: 258-65.
106. Long TC, Saleh N, Tilton RD, Lowry
GV, Veronesi B. Titanium dioxide
(P25) produces reactive oxygen species
in immortalized brain microglia
(BV2): implications for nanoparticle
neurotoxicity. Environmental Science
and Technology 2006;40:4346-52.
107. Holsapple MR, Farland WH, Landry
TD, Monteiro-Riviere NA, Carter JM,
Walker NJ et al. Research strategies for
safety evaluation on nanomaterials, part
II: toxicological and safety evaluation
on nanomaterials, current challenges
and data needs. Toxicological Sciences.
2005;88: 12-7.
108. Magrini A, Bergamaschi A, Bergamaschi
E. Nanotubi di carbonio (Ntc) e
nanoparticelle (Np): interazione con
i sistemi biologici con particolare
riferimento all’apparato respiratorio. G
Ital Med Lav Eng. 2006;28:266-9.
109. Oberdòrster G, Maynard A, Donaldson
K, Castranova V, Fitzpathck J, Ausman
K et al. Principles for characterizing
the potential human health effects from
exposure to nanomaterials: elements of
a screening strategy. Part Fibre Toxicol.
110. Oberdòrster G, Sharp Z, Atudorei V,
Elder A, Gelein R, Kreyling W et al.
Translocation of inhaled ultrafine particles
to the brain. Inhal Toxicol. 2004;16:43745.
111. Lockman RR, Mumper RJ, Khan MA,
Alien DD. Nanoparticle technology for
drug delivery across the blood-brain
barrier. Drug Dev Ind Pharm. 2002;28:112.
112. Shvedova AA, Castranova V, Kisin
ER, Schwegler-Berry D, Murray AR,
Gandelsman VZ et al. Exposure to
carbon nanotube material: assessment
of nanotube cytotoxicity using human
keratinocyte cells. J Toxicol Environ
Health A 2003;66:1909-26.
113. Rahman Q, Lohani M, Dopp E, Pemsel
H, Jonas L, Weiss DG et al. Evidence
that ultrafine titanium dioxide induces
micronuclei and apoptosis in Syrian
hamster embryo fibroblasts. Environ
Health Perspect. 2002;110:797-800.
114. Hansen T, Clermont G, Alves A, Eloy
R, Brochhausen C, Boutrand JP et
al. Biological tollerance of different
materials in bulk and nanoparticulate
form in a rat model: Sarcoma development
by nanoparticles. J.R. Cos. Interface
115. Ballestri M, Baraldi A, Gatti AM, Furci
L, Bagni A, Loria P et al. Liver and
kidney foreign bodies granulomatosis in
a patient with malocclusion, bruxism and
worn dental prosthesis. Gastroenterology
116. Hansen T, Clermont G, Alves A, Eloy
R, Brochhausen C, Boutrand JP et al.
Biological tollerance of different materials
in bulk and nanoparticulate form in a
rat model: Sarcoma development by
nanoparticles. J.R. Cos. Interface 2006;
Water and Soil Monitoring for the Protection of Environment and Human Health
117. Gatti AM, Kirkpatrick J, Gambarelli
A, Capitani F, Hansen T, Heloy R et al.
ESEM evaluation of muscle/nanoparticles
interface in a rat model. Mater Sci Mater
Med 2008;19(4):1515-22.
118. Gatti AM, Balestri M, Bagni A.
Granulomatosis associated to procelain
wear debris. American Journal of
Dentistry 2002;15(6):369-372.
119. Gatti AM, Tossini D, Gambarelli A,
Montanari S, Capitani F. Investigation
of the Presence of Inorganic Micro- and
Nanosized Contaminants in Bread and
Biscuits by Environmental Scanning
Electron Microscopy. Crit Rev Food Sci
Nutr. 2009;49(3):275-82.
120. Guildford AL, Poletti T, Osbourne
LH, Di Cerbo A, Gatti AM, Santin
M. Nanoparticles of a different source
induce different patterns of activation in
key biochemical and cellular components
of the host response. J R Soc Interface
121. Bregoli L, Chiarini F, Gambarelli
A, Sighinolfi G, Gatti AM, Santi P
et al. Toxicity of antimony trioxide
nanoparticles on human hematopoietic
progenitor cells and comparison to cell
lines. Toxicology 2009;262(2):121-9.
122. Moore MN. Do nanoparticles present
ecotoxicological risks for the health of
the aquatic environment? Environment
International 2006;32:967– 976.
123. Tang CY, Shang Fu Q, Robertson
AP, Croddle CS, Leckie JO. Use of
reverse osmosis membranes to remove
perfluorooctane sulfonate (PFOS) from
semiconductor wastewater. Environ Sci
Technol 2006;40:7343-7349.
124. Hekster F, Laane RWPM, de Voogt P.
Environmental and toxicity effects of
perfluoroalkylated substances. Rev.
Environ. Contam. Toxicol. 2003;179:99–
125. Andersen ME, Butenhoff JL, Chang
A-C, Farrar DG, Kennedy GL, Lau C
et al. Perfluoroalkyl acids and related
chemistries – Toxicokinetics and modes
of action. Toxicological Sciences 2008;
126. Condor JM, Hoke RA, de Wolf W,
Russell MH, Buck RC. Are PFCAs
bioaccumulative? A critical review and
comparison with regulatory criteria and
persistent lipophilic compounds. Environ
Sci Technol 2008;42:995–1003.
127. Prevedouros K, Cousins IT, Buck RC,
Korzeniowski SH. Sources, Fate and
Transport of Perfluorocarboxylates.
Environ Sci Technol 2006;40:32-44.
128. McLachlan MS, Holmstrom KE, Reth
M, Berger U. Riverine Discharge of
Perfluorinated Carboxylates from the
European Continent. Environ Sci Technol
129. Jone PD, Hu W, de Coen W, Newsted JL,
Giesy JP. Binding of perfluorinated fatty
acids to serum proteins. Environ. Toxicol.
Chem. 2003;22:2639-2649.
130. Martin JW, Whittle DM, Muir DCG,
Mabury SA. Perfluorinated contaminats
in a food web from Lake Ontario. Environ
Sci Technol 2004;38:5379-5385.
131. Kannan K, Tao L, Sinclair E, Pastva
SD, Jude DJ, Giesy JP. Perfluorinated
compounds in aquatic organisms at
various trophic levels in a Great Lake
food chain. Arch Environ Contam Toxicol
2005; 48:559-566.
132. Ellis DA, Martin JW, De Silva AO,
Mabury SA, Hurley MD, Sulbaek A
et al. Degradation of fluorotelomer
alcohols: a likely atmospheric source of
perfluorinated carboxylic acids. Environ
Sci Technol 2004;38:3316-3321.
133. Loos R, Locoro G, Huber T, Wollgast J,
Christoph E, de Jager AM et al. Analysis
of perfluorooctanoate (PFOA) and
other perfluorinated compounds (PFCs)
in the River Po watershed in N-Italy.
Chemosphere 2008;71:306–313.
134. Samailoff MR, Bell J, Birkholz DA,
Webster GRB, Arnott EG, Pulak R et al.
Environmental Science and Technology
135. Brack W, Klamer HJC, Lopez M, Barcelo
D. Effect-directed analysis of key toxicants
in European river basin a review. Env Sci
Pollut Res 2007;14(1):30-38.
CNR Environment and Health Inter-departmental Project
136. Brack W, Schmitt-Jansen M, Machala
M, Brix R, Barcelo D, Schymenski E et
al. How to confirm identified toxicants
in effect-directed analysis. Anal Bioanal
Chem 2008;390:1959-1973.
137. Viganò L, Mandich A, Benfenati E,
Bertolotti R, Bottero S, Porazzi E et
al. Investigating the estrogenic risk
along the river Po and its intermediate
section. Arch. Environ. Contam. Toxicol.
138. Keiter S, Grund S, Van Bavel B, Hagberg
J, Engwall M, Kammann U et al. Activities
and identification of aryl Hydrocarbon
receptor agonists in sediment from the
Danube river. Analytical and Bioanalytical
Chemistry 2008;390:2009-2019.
Role of Atmospheric Pollution on Harmful
Health Effects
A. Pietrodangeloa, M. Bencardinoa, A. Cecinatoa, S.
Decesarib, C. Perrinoa, F. Sprovieria, N. Pirronea and
M.C. Facchinib
a. CNR, Institute of Atmospheric Pollution Research (IIA) Monterotondo
St. (Roma), Italy
b. CNR, Institute of atmospheric sciences and climate (ISAC), Bologna, Italy
[email protected]
Gaseous and particulate matter in ambient and indoor air has a key role on the increased morbidity or mortality
observed in many clinical studies. Knowledge of the main toxicity patterns of atmospheric pollutants is
still at an initial stage, especially as concerns particulate matter. This is mainly due to the varying sizedistributions and chemical composition of PM10 and PM2.5 and to the many-sided toxicity mechanisms
of ultrafine particles (UFPs). In this paper, recent findings on toxicity routes attributable to PM matter (i.e.
the water-soluble organic fraction (WSOC), studied for the strong oxidative potential to biological tissues),
to UFPs and to gases, are reviewed. Toxicity routes are discussed as evidence or hypothetical relationships
between sources, diffusion paths, receptor sites and susceptible populations. Finally, strategic points are
underlined which will be further developed in the “Pilot study for the assessment of health effects of the
chemical composition of ultrafine and fine particles in Italy” project.
Adverse health effects of atmospheric
pollutants have been well documented in
Europe and in other parts of the world.
These include many diseases and an
estimated reduction of a year or more in life
expectancy for people living in European
cities. There is also evidence of increased
infant mortality in highly polluted areas.
Concerns about these health effects have
led to the implementation of regulations
to reduce harmful air pollutants emissions
and their precursors at international,
national, regional and local levels. Further
measures – while necessary to further
reduce the health effects of air pollution
– are becoming increasingly expensive.
There is thus a growing need for accurate
information on the health effect of air
pollution to plan scientific, effective and well
targeted strategies and reduce these effects.
In July 2002 the European Parliament
and the Council adopted the Decision
1600/2002/EC on the Sixth Community
Environment Action Programme (Sixth
EAP). This Programme sets out the key
environmental objectives to be attained in
the European Community, one of which
(Article 2) is to establish “. a high level
of quality of life and social well being
for citizens by providing an environment
where the level of pollution does not give
rise to harmful effects on human health
…”(1). The activities of the European
Commission to implement the Sixth EAP
currently take place within the Clean Air
for Europe (CAFE) programme (2). This
programme, launched in early 2001, aims
at developing long-term, strategic and
integrated policy advice to protect against
significant negative effects of air pollution
on human health and the environment.
The World Health Organization (WHO)
in support to the CAFE process, provided
updated information on health effects
CNR Environment and Health Inter-departmental Project
of air pollutants establishing the project
“Systematic Review of Health Aspects
of Air Quality in Europe” (3) in the
course of which the current state of
knowledge concerning health impacts
of air pollution has been reviewed. The
body of evidence of air pollution effects
on health at the pollution levels currently
common in Europe has been considerably
strengthened by the contribution of
both epidemiological and toxicological
studies. The latter provide new insights
into possible mechanisms to analyse the
hazardous effects of air pollutants on human
health and complement the large body of
epidemiological evidence, showing, for
example, consistent associations between
daily variations in air pollution and some
health outcomes. Exposure to ambient air
pollution has been linked to a number of
different health outcomes, starting from
modest transient changes in the respiratory
tract and impaired pulmonary function, to
restricted activity/reduced performance,
emergency room visits and hospital
admissions and mortality. There is also
increasing evidence of air pollution adverse
effects not only on the respiratory system,
but also on the cardiovascular system.
This evidence stems from studies on both
acute and chronic exposure. Short-term
epidemiological studies suggested that
a number of sources are associated with
health effects, especially motor vehicle
emissions, and also coal combustion.
These sources produce primary as well
as secondary particles, both of which
have been associated with adverse
health effects. If long-term exposure to a
specific pollutant is linked to some health
effects, cohort studies provide a basis to
estimate chronic diseases and lifespan
reduction in a given population. This is
the case for mortality linked to PM longterm exposure. An expert group led by
Task Force on Health Aspects of Long
Range Transboundary Air Pollution –
recommended the use of risk coefficients
from the American Cancer Society (ACS)
study (4) to estimate the effects of chronic
exposure to particulate matter (PM) on
life expectancy in Europe. This study is
the largest cohort study published in the
scientific literature on the association
between mortality and exposure to PM
in air, and has involved 550,000 persons
between 1982 and 1998. The risk estimates
from this study were also used in the WHO
Global Burden of Disease project (5). This
project estimated that exposure to fine
PM in outdoor air leads to about 100,000
deaths and 725,000 years of life lost each
year in Europe. It is clear that there is a
significant health risk associated with PM.
It is also clear that there is a yet not known
safe threshold for exposure but that there
appears to be a linear relationship between
exposure and risk. In addition, it has not yet
been possible to identify with confidence
which PM chemical constituents are
primarily responsible for the different
health effects. Therefore, even though
the evidence on the relationship between
exposure to different air pollutants and
health effects has increased considerably
over the past few years, there are still
large uncertainties and important gaps in
knowledge. These gaps can be reduced
only by targeted scientific research. Areas
in which such research is urgently needed
include exposure assessment, dosimetry,
toxicity of different components, biological
mechanisms of effects, susceptible groups
and individual susceptibility (taking into
account gene–environment interactions),
effects of mixtures versus single substances,
and effects of long-term exposure to air
pollution. The “Systematic Review” clearly
demonstrated the need to set up a more
Role of Atmospheric Pollution on Harmful Health Effects
comprehensive air pollution and health
monitoring and surveillance programme
in different European cities. Air pollutants
to be monitored include coarse PM, PM2.5,
PM1, ultrafine particles, PM chemical
composition, including elemental and
organic carbon, and gases such as ozone,
nitrogen dioxide and sulphur dioxide. The
value of black smoke and ultrafine particles
as indicators of traffic-related air pollution
should also be evaluated. Furthermore,
periodic surveillance of health effects
requires better standardization of
routinely collected health outcome data.
The “Systematic Review” also showed the
need of a system to maintain the literature
database and develop meta-analysis to
monitor research findings, summarize
the literature on health effects and health
impact assessment.
Ambient air pollution consists of a
highly variable, complex mixture of
different substances, which may occur
in the gas, liquid or solid phase. Several
hundred different components have been
found in the troposphere, many of them
potentially harmful to human health and
the environment. The main sources of air
pollution are transport, power generation,
industry, agriculture, and heating. All these
sectors release a variety of air pollutants
– sulphur dioxide, nitrogen oxides,
ammonia, volatile organic substances,
and particulate matter – many of which
interact with others to form new pollutants.
These are eventually deposited and have a
whole range of effects on human health,
biodiversity, buildings, crops and forests.
Air pollution results in several hundreds of
thousands of premature deaths in Europe
each year, increased hospital admissions,
extra medication, and the loss of millions
of working days. The health costs for the
European Union are huge. The pollutants
of highest concern for human health are
airborne particulates and ozone – indeed
no safe levels have yet been identified
for either of them. Nevertheless, the
“Systematic Review” focused on three
pollutants: particulate matter (PM), ozone
and nitrogen dioxide, as requested by
the CAFE Steering Group. This is not to
imply that other substances do not pose a
considerable threat to human health and the
environment at the current levels present
in Europe. It should also be mentioned
that PM itself is a complex mixture of
solid and liquid constituents, including
inorganic salts such as nitrates, sulphates
and ammonium and a large number of
carbonaceous species (elemental carbon
and organic carbon). Thus PM implicitly
covers a number of different chemical
pollutants emitted by various sources. The
term ‘particulate matter’ (PM) is used to
describe airborne solid particles and/or
droplets. These particles may vary in size,
composition and origin. Several different
indicators have been used to characterize
ambient PM. Classification by size is quite
common because size governs the transport
and removal of particles from the air and
their deposition within the respiratory
system, and is at least partly associated
with the chemical composition and
sources of particles. Based on size, urban
PM tends to be divided into three main
groups: coarse, fine and ultrafine particles.
The border between the coarse and fine
particles usually lies between 1 μm and
2.5 μm, but is usually fixed by convention
at 2.5 μm in aerodynamic diameter (PM2.5)
for measurement purposes. The border
between fine and ultrafine particles lies
at about 0.1 μm. PM10 is used to describe
particles with an aerodynamic diameter
smaller than 10 μm. The particles
CNR Environment and Health Inter-departmental Project
Figure 1. Deposition probability of inhaled
particles in the respiratory tract according
to particle size.
contained in the PM10 size fraction may
reach the upper part of the airways and
lung. Fig. 1 shows schematically where
particles are deposited in the respiratory
tract, depending on their size.
Smaller particles (in particular PM2.5)
penetrate more deeply into the lung and
may reach the alveolar region. Ultrafine
particles contribute only slightly to PM10
mass but may be important from a health
point of view because of their large
numbers and high surface area. They are
produced in large numbers by combustion
(especially internal combustion) engines.
As reported, (3) the most severe effects in
terms of the overall health burden include
a significant reduction, by a year or more,
in average life expectancy linked to the
long-term exposure to high levels of air
pollution due to fine PM. Many studies
have found that fine particles (usually
measured as PM2.5) have serious effects on
health, such as increased mortality rates
and emergency hospital admissions for
cardiovascular and respiratory reasons.
Thus there is good reason to reduce
exposure to such particles. Coarse particles
(usually defined as the difference between
PM10 and PM2.5) seem to have effects
on, for example, hospital admissions for
respiratory illnesses, but their effect on
mortality is less clear. Nevertheless, there
is sufficient concern to consider reducing
exposure to coarse particles as well as to
fine particles. Similarly, ultrafine particles
are different in composition, and probably
to some extent in effect, from fine and
coarse particles. Nevertheless, their effect
on human health have been insufficiently
studied to permit a quantitative evaluation
of health risks due to exposure to such
As stated above, PM in ambient air has
various sources. In targeting control
measures, it would be important to know
if PM from some sources or of a specific
composition gave rise to special health
concern due to their high toxicity. The few
epidemiological studies that have addressed
this important issue specifically suggest
that combustion sources are particularly
important for health. Toxicological studies
have also pointed to primary combustionderived particles as having a higher toxic
potential. These particles are often rich in
transition metals and organic compounds,
and also have a relatively high surface
area. By contrast, several other single
components of the PM mixture (e.g.
ammonium salts, chlorides, sulphates,
nitrates and wind-blown dust such as
silicate clays) have been shown to have a
lower toxicity in laboratory studies.
Despite these differences found among
the constituents studied in laboratory, it
is currently not possible to quantify the
contributions of the different sources and
different PM components on the health
effects caused by exposure to ambient
2.1. Modelling approach on health impact
Health impact assessment allows to
quantify the effects of exposure to an
environmental hazard. It plays a central
Role of Atmospheric Pollution on Harmful Health Effects
role in assessing the potential health effects
of different policies and measures, thereby
providing a basis for decision-making. A
detailed knowledge of several factors is
required for any such assessment. Crucial
information on exposure to air pollutants
is provided by an integrated approach
on ambient air quality monitoring and
modelling study.
Air quality modelling, particularly the
Integrated Assessment Modelling (IAM),
is important in linking pollution levels
to emission sources and integrating
epidemiological studies, information
about the formation and dispersion of fine
particles in the atmosphere, assessment
of current and future levels of emissions
of fine particles and their precursors. In
the frame of the UN-ECE Convention on
Long-Range Transboundary Air Pollution
(CLRTAP), and in the context of the
Community Environmental policies of
the EU Commission, the RAINS-Europe
model provides one of the most relevant
examples of successful application of
Integrated Assessment Modelling (IAM).
The RAINS model (6), developed at
the International Institute for Applied
Systems Analysis (IIASA), considers
emissions of SO2, NOx, PM10, PM2.5,
VOC and NH3, provides deposition and
concentration maps and addresses threats
to human health posed by fine particulates
(7). The assessment of fine particle health
impacts is implemented through the Life
Expectancy Reduction indicator (LER),
defined as months lost attributable to
PM2.5 concentrations. Awaiting further
refinements in the scientific disciplines,
the quantitative implementation should be
considered as preliminary and needs to
be revised as soon as more substantiated
scientific information becomes available.
The Task Force on Health of the United
Nations Economic Commission, when
conducting the in-depth review of the
RAINS approach for modelling health
impacts of fine particles (TFH, 2003),
noted “that some data suggested that
different components that contributed
to PM2.5 mass might not be equally
hazardous. In particular, the discussion
focused on the role of the secondary
inorganic aerosols (including nitrates and
sulphates). It concluded that, due to the
absence of compelling toxicological data
about the active different PM components
of a complex mixture, it was not possible
to quantify the relative health impact
importance of the main PM components at
this stage”. Therefore, it was recommended
Figure 2. Changes in EU life expectancy loss in 2000 and in the interim objective in 2020
(Strategy) (9).
CNR Environment and Health Inter-departmental Project
to relate health impacts to total mass of
PM2.5 until more specific evidence becomes
available (8). The methodology used in the
RAINS model, at European and national
scale, to estimate losses in life expectancy
due to air pollution represents an initial
implementation assessing the implications
of present and future European policies to
control exposure to particulate matter. In
the Figure 2, an example of the changes
in life expectancy loss in EU in 2000 and
in the interim objective in 2020 (9) are
reported. The impact assessment of the
different policies is based on the analysis
of a set of technological measurements
with the RAINS model related to various
emission reduction scenarios. The
ambition level of the Strategy is based on a
set of specific measurements which would
need to be undertaken at Community and
Member State level.
In the recent years, some European
countries such as, among others, Italy,
have tackled the issue of implementing
the RAINS model at national level,
introducing higher spatial resolution in
similar models, pursuing the ultimate
objective of a more adequate response to
the need of evaluating, at national level,
cost-effective policy measures to reduce
air pollutant emissions, and consequently,
the pressure on environment and human
health. As a result, the RAINS-Italy model
(10) as the national version of the RAINSEurope model was defined considering as
emission source areas either the nation as a
whole or the 20 administrative Regions. In
a recent work (11), the RAINS-Italy model
Figure 3. Losses in average Life Expectancy (months) attributable to PM2.5 concentrations
at 2010: a) CLE scenario; b) difference (months) between Air Quality Management Plan
scenario and CLE scenario (11).
Role of Atmospheric Pollution on Harmful Health Effects
was used to assess the emission reduction
strategies followed in the Regional Air
Quality Management Plans (AQMPs) to
meet environmental quality targets by
means of Technical and Non-Technical
Measures. Regarding health impacts
(Figure 3a), the most important Italian
metropolitan and industrial areas show
an average Life Expectancy loss ranging
between 12 to 23 months in the 2010-CLE
scenario. This higher resolution map shows
a better definition of the hot spots present
in the urban areas of Turin, Milan, Rome
and Naples, as well as in the industrial sites
of eastern Sicily and Taranto, in the Apulia
region (12). The above mentioned study
showed that if compared to the 2010 CLE
scenario, the 2010 AQMP scenario reduces
PM10, NOx and SO2 emission by 2.8%, 2.4%
and 0.5%, respectively. Regions with a
more effective AQMP reach higher PM10
yearly average concentrations reductions,
with peaks of 7.5% in northern and central
Italy, even if this is not sufficient to assure
the compliance with air quality standard
in 2010. Similarly the improvement in the
average Life Expectancy loss indicator
(Figure 3b) is of 1 month only in Lombardy
2.2. Indoor vs outdoor air pollution
As it is the case for other air pollutants, the
total exposure of an individual to suspended
particulate matter (of whatever size) is
the result of contributions from the two
microenvironments, outdoor air and indoor
air. The indoor air compartment can be
further subdivided into homes, restaurants,
car, buses and aircraft, workplaces etc.
Consequently, in studies to detect and
quantify the health effects of particles,
attention must be paid that exposure is
characterised adequately. Generally there
are two different ways to obtain such
characterization. One is by measuring
total air exposure using personal sampling:
the persons under study are provided each
with a personal sampler that they have to
carry on them or position as close to them
as possible for 24 hours consecutively.
Since this is cumbersome for a study
participant, the following alternative
can be used: total exposure is modelled
taking into account the time spent in the
various microenvironments (indoors and
outdoors) and the concentrations observed
in these microenvironments. Personal
sampling provides a concentration level
that represents the integration of all the
concentration levels in all compartments
visited by the studied person during the
24-h (or longer) measurement period
and, thus, it cannot detect the individual
contribution of any compartment. In
contrast, the modelling process using the
combination of the pollutant concentrations
in the different microenvironments and
the time spent therein permits to assess
the contribution of total exposure to each
of these microenvironments. This kind
of source apportionment can be of great
help to decide what measurements should
receive priority in controlling pollutant
concentrations. A recent publication on
exposure to PM2.5 describing the results
of a model approach (13) stated that the
had the greatest influence on total
exposure to PM2.5, compared to the other
microenvironments considered, namely
outdoor and non-residential indoor (office,
school, store, restaurant, bar, in-vehicle). It
turned out that the outdoor compartment
was responsible for a direct contribution of
about 5% on average. Another 35% was due
to an indirect contribution via infiltration of
outdoor air into indoor spaces. Thus, about
60% of the total exposure to PM2.5 could
not be influenced by control measurements
taken to reduce outdoor air PM2.5 levels.
CNR Environment and Health Inter-departmental Project
3.1 Gaseous matter
Air pollution by inorganic gaseous matter
was dealt with since the first major
pollution events (i.e. the Great Smog
of London, 1952; etc.) put on evidence
the strict relationship between levels of
chemical species in the ambient (and
indoor) air their harmful effects on
health and ecosystems. Among inorganic
gases, carbon monoxide (CO) is one of the
most common air pollutants. It has a low
reactivity and a low water solubility and
it is mainly released into the atmosphere
as a product of incomplete combustion.
CO is not only directly released in the air,
but can also originate from the chemical
reactions of organic air pollutants, such
as methane. Its latency in the atmosphere
is about three months. Since at moderate
latitudes air masses travel for months and
since the CO formation from organic air
pollutants takes place everywhere in the
atmosphere, a global background level
of CO exists, ranging between 0.05 and
0.15 ppmv (0.06 and 0.17 mg/m3) (14). It
is estimated that about one-third of CO,
including that derived from hydrocarbon
oxidation, originates from natural sources.
CO levels in busy city streets are higher
than those present near highways, since
the amount of CO emissions per kilometre
strongly decreases with vehicle speed and
also because ventilation in city streets
is less. Ambient CO levels are usually
highest in winter, because cold engines
release much more CO than hot engines
and also because the atmosphere tends
to be more stable than in summer. It has
to be reminded, however, that usually
CO ambient levels do not exceed neither
WHO guidelines for health protection
nor the limits of the EU directives on air
quality. Although CO is hardly removed
from the air in atmospheric transport at
continental level, long range transport
does not lead to concentrations of concern
at both rural and urban background level.
Also at points of high traffic in large cities,
levels exceeding legislation are only
occasionally observed. Industrial areas
may be affected by large CO emissions;
however, when these emissions are released
through high chimneys, local ambient
concentrations show poor increases and
do not pose risks for human health14. CO
toxicity patterns are linked to its reaction
with haemoglobin in the human blood to
form carboxyhaemoglobin (COHb). The
affinity of haemoglobin for CO is 200250 times higher than for oxygen, and as
a result this binding reduces the oxygencarrying capacity of the blood and impairs
the release of oxygen to extra vascular
tissues. The most important variables
determining the COHb level are CO in
inhaled air, duration of exposure and lung
ventilation. Physical exercise accelerates
the CO uptake process.
The formation of COHb is a reversible
process; however, the half-life elimination
of COHb is much longer in the foetus than
in the pregnant mother. The effects of CO
exposure on cardiovascular disease have
been studied for a long time (15). However,
only limited information is available about
the possible cardiac effects of gaseous
pollutants at concentrations close to those
present in ambient air. Apart from hazards
due to high CO concentrations, other
health effects seem to originate from the
association between CO and other gaseous
and particulate matter, especially exposure
to exhausts from motor engines. Although
attention has recently been focused on the
cardiovascular effects of PM, few studies
show evidence of the relationship between
some cardiovascular diseases and the
exposure of different populations to road
Role of Atmospheric Pollution on Harmful Health Effects
traffic exhausts. For example, experimental
studies have demonstrated mild cardiac
effects from both sulphur dioxide (SO2)
and ozone (O3)(16). Other studies, where
personal exposure to different pollutants
has been investigated, have suggested that
the estimated cardiac effects attributed to
gases, including SO2, are actually effects
of other pollutants, specifically PM (17).
At this stage of knowledge, however, it
is difficult to differentiate between the
effects of PM and those of gases because
people are normally exposed to both types
of pollutants at the same time. Because
of these uncertainties, it seems prudent
to further investigate both the effects that
low concentrations of gaseous pollutants,
alone or in combination with PM, might
have on cardiovascular diseases, and
the possibility that the associations with
gaseous pollutants may actually reflect the
effects of PM or some component that is
not currently being studied for its health
effects (16).
Anthropogenic sulphur dioxide (SO2)
results from the combustion of sulphurcontaining fossil fuels (mainly coal and
heavy oils) and the smelting of sulphur
containing ores. Over the past years,
however, there has been a net tendency
towards emission reduction in Countries
where low-sulphur fuels and emission
control measures have been adopted. In
addition, the source pattern has changed
and moved from small multiple sources
(domestic, commercial, industrial) to
large single sources releasing SO2 from
tall stacks. Volcanoes and oceans are the
major natural sources of SO2. After being
released in the atmosphere, sulphur dioxide
is further oxidized to sulphate (SO4=) and
sulphuric acid forming an aerosol often
associated with other pollutants in droplets
or solid particles having a wide range of
sizes. SO2 and its oxidation products are
removed from the atmosphere by wet
and dry deposition. Nowadays, it is also
recognized that sulphate aerosols play
an important cooling role in the radiative
climate of the Earth through the phenomena
of sunlight scattering in cloud free air and
as cloud condensation nuclei. Sulphur
dioxide is an irritant and when inhaled at
high concentrations may cause breathing
difficulties in people exposed to it. People
suffering from asthma and chronic lung
disease may be especially susceptible to
the adverse effects of sulphur dioxide.
Nevertheless adverse effects from high
concentrations of SO2 have been observed
both on healthy people and asthma patients
Oxidized nitrogen compounds (NO2, NOx,
NOy) and ozone (O3) join common patterns
in atmospheric formation chemistry,
environmental fate and adverse effects on
health and the ecosystem. NO is directly
released by all combustion processes; once
in the atmosphere, it reacts with oxygen
and a number of other inorganic (e.g. O3,
OH radical, halogens) and organic (VOCs)
gases to form NO2, NO3, HONO, HNO3,
PAN, nitro – PAH and other organic
and halogen nitrates, in the gaseous or
particulate phase. Ozone and oxidized
nitrogen compounds are strongly oxidant
and this aspect mainly characterizes their
harmful health action. In particular, the
oxidizing potential of these compounds
is commonly referred to as “odd oxygen”
(Ox) or “odd nitrogen” (NOx), i.e families
of chemical compounds that interconvert
rapidly among themselves on time scales
that are shorter than those necessary to
form or destroy the family . Another family
is that defined as “NOz”, which refers to
the sum of NOx oxidation products (19).
NOx = NO + NO2 [1]
CNR Environment and Health Inter-departmental Project
Ox = Σ (O(3P)
+ O(1D) + O3 + NO2) [2]
NOz = Σ (HNO3 + HNO4 + NO3 + 2NO2O5
+ PAN + other organic nitrate + halogen
nitrate + particulate nitrate) [3]
Unlike some other compounds whose
formation rates vary directly with the
emissions of their precursors, O3 differs
in that its production changes nonlinearly
with the concentrations of precursors.
At the low NOx concentrations found in
most environments ranging from remote
continental areas to rural and suburban
areas, the O3 net production increases
with the increasing of NOx. At the high
NOx concentrations found in downtown
metropolitan areas especially near busy
streets and roadways and in power plants,
there is a net destruction of O3 by titration
reaction with NO. Between these two
regimes is a transition stage in which O3
shows only a weak dependence on NOx
concentrations. In the high NOx regime,
NO2 scavenges OH radicals which would
otherwise oxidize VOCs to produce peroxy
radicals, which in turn would oxidize NO
into NO2. In the low NOx regime, VOC
oxidation generates, or at least does not
consume, free radicals, and O3 production
varies accordingly. Sometimes the terms
‘VOC-limited’ and ‘NOx-limited’ are
used to describe these two regimes; also,
the terms NOx-limited and NOx-saturated
are used. The chemistry of OH radicals,
that are responsible of the initiation of
hydrocarbons oxidation, shows a behaviour
similar to that of O3 with respect to NOx
concentrations (19). These considerations
introduce a high degree of uncertainty
into attempts to relate changes in O3
concentrations to precursors emissions. It
should also be noted at the outset that in
a NOx-limited (or NOx-sensitive) regime,
O3 formation is not insensitive to radical
production or the flux of solar UV photons,
but O3 formation is more sensitive to NOx.
For example, global tropospheric O3 is
sensitive to CH4 concentrations even if the
troposphere is predominantly NOx-limited.
To get information about the O3-NOx-VOCs
relationships and sensitivity, the ratio
of summed VOC to NOx concentrations
determining whether conditions are NOxsensitive or VOC sensitive is not sufficient
to describe O3 formation, since other
factors - i.e. the effect of biogenic VOCs
(which are not present in urban centres
in early morning) - and some important
individual differences in VOCs ability
to generate free radicals, have to be
considered. The difference between NOxlimited and NOx-saturated regimes is also
reflected in measurements of hydrogen
peroxide (H2O2), another strong oxidant of
ambient air. H2O2 formation takes place by
self-reaction of photo chemically generated
HO2 radicals, so that there is large seasonal
variation in H2O2 concentrations, and
values in excess of 1 ppb are mainly limited
to summer months, when photochemistry
is more active (20). Hydrogen peroxide is
produced in abundance only when O3 is
produced under NOx-limited conditions.
The transition from NOx - limited to NOx
- saturated conditions is highly space and
time dependent. In the upper troposphere,
responses to NOx additions from commercial
aircraft have been found that are very
similar to those in the lower troposphere.
Moreover, the complex interplay between
chemical and meteorological processes
gives rise to uncertainties in understanding
ozone formation. This is especially true for
regions of complex topography. In coastal
regions around the Mediterranean Basin,
for instance, the combination of mountain
and sea breeze re-circulations significantly
affects ozone phenomenology. Ozone
can also have very specific distributions
Role of Atmospheric Pollution on Harmful Health Effects
in mountain areas, and observed
concentrations differ significantly between
mountain peaks and valleys (20). Nitropolycyclic aromatic hydrocarbons (nitroPAHs) are generated from incomplete
combustion processes through PAHs
electrophilic reactions in the presence
of NO2 (21). Among combustion sources,
diesel emissions have been identified as the
major source of nitro-PAHs in ambient air.
Direct emissions of nitro-PAHs in PM vary
with the type of fuel, vehicle maintenance,
and ambient conditions (22). In addition
to being directly released, nitro-PAHs can
also be formed from both PAHs gaseous
and heterogeneous reactions with gaseous
nitrogenous pollutants in the atmosphere.
After formation, nitro-PAHs with low
vapour pressures (such as 2NF and 2NP)
immediately migrate to particles under
ambient conditions; therefore harmful
effects related to nitro – PAHs are better
investigated in the organic fraction of
particulate matter. An extended discussion
on this topic is reported in par. 3.2.
Also in indoor environments NO2 plays
a key role in adverse health effects. It is
indeed produced by NO reactions with
ozone or peroxy radicals generated
by indoor air chemistry involving O3
and unsaturated hydrocarbons such as
terpenes found in air fresheners and other
household products (23). Nevertheless
indoor NO2 is also contributed by indoor
– outdoor air exchange. The relationship
between personal NO2 exposure and
ambient NO2 can be modified by the
indoor environment. For example, during
the infiltration processes, ambient NO2
can be lost through penetration and decay
(chemical and physical processes) in the
indoor environment, and the concentration
of indoor ambient NO2 is not just the
ambient NO2 concentration but the product
of the ambient NO2 concentration and the
infiltration factor (Finf, or α if people spend
100% of their time indoor). Indoor NO2
is removed by gas phase reactions with
ozone and assorted free radicals and by
surface promoted hydrolysis and reduction
reactions. The concentration of indoor
NO2 also affects PAN decomposition.
These processes are important not only
because they influence the indoor NO2
concentrations to which humans are
exposed, but also because some products
of indoor chemistry may confound
attempts to examine associations between
NO2 and health. As a matter of fact, NO2
is an oxidant and lipid peroxidation is
believed to be a major molecular event
responsible for its toxicity. As a result,
there has been considerable attention paid
to NO2 effect on the antioxidant defence
system in the epithelial lining fluid and
in pulmonary cells. Repeated exposure to
indoor NO2 at concentrations ranging from
0.04 to 33 ppm has been shown to alter
low molecular weight antioxidants such
as glutathione, vitamin E, and vitamin
C, as well as some enzymes involved in
cell oxidant homeostasis. NO2 effects
on structural proteins of the lungs have
raised concern because elastic recoil is
lost after exposure. It has been observed
that the latter increases collagen synthesis.
This, in turn, shows increases in total
lung collagen which, if sufficient, could
result in pulmonary fibrosis after longer
periods of exposure. Such correlation has
yet to be confirmed by in vivo studies
involving NO2 exposure; nevertheless
some evidence shown in animal studies
about asthma, emphysema and other lung
diseases. Similarly to O3, NO2 is absorbed
throughout the lower respiratory tract, but
the major delivery site is the centriacinar
region, i.e, the junction between the
conducting and respiratory airways in
humans and animals (21).
CNR Environment and Health Inter-departmental Project
Ozone is a strong oxidant, and as such
can react with a wide range of cellular
components and biological materials:
damage can occur to all parts of the
respiratory tract. The time pattern of
these changes in the respiratory system,
as determined in laboratory animals as
well as in epidemiological investigations,
is complex. During the first few days of
exposure, inflammation occurs and then
persists at an attenuated level. At the same
time, epithelial hyperplasia progresses,
and reaches a plateau after about one week
of exposure. When the exposure ceases,
these effects slowly disappear. In contrast
to this, interstitial fibrosis increases slowly
and can persist even when exposure
ceases. In a large number of controlled
human studies, significant impairment of
pulmonary function has been reported.
Field studies in children, adolescents,
and young adults have indicated that
pulmonary function decrements, similarly
to those observed in controlled studies, can
occur as a result of short term exposure
to ozone concentrations in the range of
120-240 μg/m3 and higher. In comparison
with adults, children have a higher intake
of ozone and other air pollutants. This
is due to a higher basal metabolic rate,
resulting in a higher breathing volume per
minute and a higher breathing frequency.
Furthermore, their respiratory tract is still
under development until the age of six and
a half, and it is therefore more susceptible
to the inflammatory effects of ozone.
Children’s immune systems are not yet
fully developed and are generally under
bigger stress. For these and other reasons,
children are at higher risk when exposed
to ambient ozone concentrations. Hospital
admissions for respiratory causes and
exacerbation of asthma are observed both
in exposures to ambient ozone (and copollutants) and in controlled exposures to O3
alone. Other groups at risk are those people
exercising outdoors during evening hours
or whenever ozone concentrations tend to
be highest (e.g. in photochemical smog
events). Due to the irritant nature of ozone,
capable of inducing airway inflammation
and bronchoconstriction, asthma patients
are deemed to be at enhanced risk from
exposure to ozone and photochemical
smog, because inflamed airways contribute
to the pathogenesis and exacerbation of the
disease and to morbidity and mortality for
asthma. Results from recent epidemiologic
studies have suggested that ozone might
have serious cardiovascular effect (24
and references therein). Although a large
number of toxicity animal studies have
been performed on respiratory and other
effects of NO2, O3 and other gaseous
pollutants on metabolic and physiological
functions (body weight, hepatic, renal,
brain, etc.), results are often affected
by serious limitations, due to both the
necessary animal-to-human extrapolation
of concentration-response data and the fact
that controlled exposures to a single pollutant
alone provide incomplete information.
Human clinical studies attempt to recreate
in laboratory the atmospheric conditions
of ambient pollutant atmospheres, paying
great attention to concentrations, duration,
timing, and other conditions which may
impact responses. These studies allow
the measurement of health symptoms
and physiological markers resulting from
air breathing. This carefully controlled
environment allows researchers to identify
responses to individual pollutants,
relationships, to examine interactions
among pollutants, and to study the effects of
other variables such as exercise, humidity,
or temperature. Susceptible populations,
including individuals with acute and
chronic respiratory and cardiovascular
Role of Atmospheric Pollution on Harmful Health Effects
diseases, can participate with appropriate
limitations based on subject comfort and
protection from risk. Endpoint assessment
has traditionally included symptoms and
pulmonary function, but more recently
a variety of markers of pulmonary,
systemic, and cardiovascular function
have been used to assess pollutant effects.
It is reasonable to consider, however, that
human clinical studies have limitations.
For practical and ethical reasons, studies
must be limited to relatively small groups,
to short durations of exposure, and to
pollutant concentrations that are expected
to produce only mild and transient
responses. Findings from the short-term
exposures in clinical studies may provide
limited insight about the health effects of
chronic or repeated exposures. Moreover,
the choice of previous- and after-exposure
time lags for the observation of health
effects is critical in assessing the role of a
pollutant in toxicity events. Many studies
have shown that NO2 has a fairly consistent,
immediate effect on health outcomes,
including respiratory hospitalizations and
mortality. Several studies also observed
significant NO2 effects over longer
cumulative lag periods, suggesting that in
addition to single-day lags, multiday lags
should be investigated to fully capture a
delayed NO2 effect on health outcomes.
Finally it should be kept in mind that,
although many biochemical changes are
not necessarily toxic manifestations of the
pollutant per se, such changes may anyway
impact the metabolism and toxicity of
other chemicals in humans and animal
species (21).
The EU regulates the main harmful
inorganic gaseous pollutants by the EC
legislation of the Air Quality Framework
(25). Other legally binding Protocols have
been established since the 1979 Geneva
Convention on Long Range Transboundary
Air Pollution (LRTAP) (26). Guideline
levels aimed at health and environment
protection have been set by WHO and
other institutions, too, to be used for
impact assessment. The first edition of the
WHO “Air quality guidelines for Europe
(AQG)” was published in 1987 (27). To
determine critical or guideline levels,
quantitative relationships between the
pollutant exposure and its studied effect are
needed. However, any such relationships
have a certain degree of uncertainty, and
the data necessary to produce them are
often scarce. Therefore, the establishment
of guideline values, such as levels at
which acute (or chronic) effects on public
health or ecosystems are likely to be not
relevantly harmful, impose the support of
biological, clinical and epidemiological
evidence, often not available or inadequate.
Different legal tools aimed at protecting
and improving the health and quality of
ecosystems from air pollution have been
used in recent years. The 1992 fifth action
programme of the European Commission
(EAP) on the environment recommended
“the establishment of long-term air quality
objectives” for many inorganic gaseous
pollutants (CO, NO2 and NOx, SO2, O3).
The list of key requirements, also includes
the need for “studies to analyze the effects
[on health ant ecosystems] of the combined
action of various pollutants or sources
of pollution and the effect of climate
on the activity of the various pollutants
examined”. Under the 5th EAP the Air
Quality Framework was established,
within which the 96/62/EC Directive and
the following four Daughter Directives
have been adopted. This law establishes
limits and threshold values for SO2, NO2
and NOx under the 99/30/EC, CO under
the 2000/69/EC and O3 under the 2002/03/
EC for EU Member States. In the 6th EAP,
further steps have been taken toward health
CNR Environment and Health Inter-departmental Project
/ environment protection by the Clean Air
for Europe (CAFE) programme. The CAFE
is conceived as a process based on technical
analysis and policy development to achieve
the adoption of a Thematic Strategy
on Air Pollution. The major elements
of the CAFE programme are outlined
in Communication COM(2001)245 (2).
The programme, launched in early 2001,
aims at the development of a long-term,
strategic and integrated policy advice to
protect against the significant negative
effects of air pollution on human health
and the ecosystem. Within this process,
the 2008/50/EC Directive on ambient air
quality and cleaner air for Europe has been
adopted and will enter into force as from
11 June 2010, when the Directives 96/62/
EC, 1999/30/EC, 2000/69/EC and 2002/3/
EC shall be repealed.
3.2 Composition and size distribution of
the inorganic fraction of suspended PM
3.2.1 Inorganic fraction of suspended PM
The history of air pollution is very long,
and since its very first occurrence - smoke
from heating and cooking activities in
prehistoric dwellings - particles have been
addressed as one of the most important
issues. Pollution from combustion sources
and specifically suspended particles
have been responsible for the most
relevant pollution disasters (e.g. Mause
Valley, Belgium, 1930, the Big Smoke,
London, 1952), which led to increasing
efforts towards pollution monitoring,
the understanding of main pollution
processes, political awareness and, finally,
At European level, pollution from particulate
matter (PM) has been first addressed by
the First Daugther Directive (1999/30/EC)
to the Air Quality Framework Directive
(1996/62/EC); recently, a new Directive
(2008/50/EC, published in June 2008)
summarised most of the existing legislation
on ambient air and introduced some new
requirements. As far as PM is concerned,
the First Daugther Directive addressed only
PM10, setting limits for its annual average
concentration (40 μg/m3) and the number
of exceedances (35 per year) of the 50 μg/
m3 daily concentration limit. Air quality
limits also for PM2.5 were introduced
only by the recent Directive 2008/50/EC
(25 μg/m3, with a 20% margin of tolerance
that will be reduced to zero on 1st January
2015). In addition to the measurement of
PM mass concentration, Directive 2008/50/
EC also includes the measurement of PM2.5
chemical composition in background
sites, listing a number of components that
must necessarily be determined in each
PM2.5 sample (sulphate, nitrate, chloride,
sodium, ammonium, potassium, calcium,
magnesium). This new issue is related to
the increasing awareness of the complexity
of this “pollutant”, which is a mixture of
thousands of different chemical species,
each one with its own properties and
possibly its own environmental and health
Unlike gaseous pollutants, where the
concentration is generally sufficient to
define the system, for the atmospheric
aerosol many parameters have to be
defined. Physical parameters include the
geometric and aerodynamic diameter,
shape (spheres, fibres, etc.), phase
(solid, liquid, mixture of both), density,
electrical charge, hygroscopicity etc. and
are necessary to understand particles
behaviour in the atmosphere as well as
inside the respiratory system. The most
complex issue in aerosol characterisation,
however, is its chemical composition,
which includes a variety of components,
whose determination requires a variety of
analytical techniques.
Role of Atmospheric Pollution on Harmful Health Effects
The knowledge of health effects caused
by inhalation of atmospheric particles has
been improved a lot during the last decade
and there is no doubt that particles can be
harmful to human health. PM is associated
with a wide variety of both acute and
chronic cardiovascular and respiratory
effects. Acute effects include increased
hospital admittance for respiratory disease
or premature mortality for cardiovascular
disease, while chronic effects include a
number of diseases leading to longevity
reduction. The increase in respiratory and
cardiovascular morbidity and mortality
is in the order of a few percent for a PM
increase of 10 μg/m3 (28-34).
The study of the link between particulate
matter and health is extremely complex
and poses many problems, including
the difficulties in assessing the role of
particle size and particle composition,
in quantifying the real exposure and
understanding the biological mechanisms
that are responsible for the effects, in
evaluating the impact of the different
sources and, last but not least, in detecting
the real concentration and composition of
the atmospheric aerosol.
The size of atmospheric particles varies
among five orders of magnitude, from a few
nanometres to hundreds of micrometers.
The size of the aerosol influences its lifetime
in the atmosphere (and thus the spatial
range of influence of any single source)
as well as its pathway inside the human
body. Basically, the atmospheric aerosol
consists of three modes, which are closely
linked to their formation mechanism: the
coarse mode, predominantly mechanically
generated (e.g. by erosion and by resuspension), the accumulation mode,
produced by condensation from vapours
and coagulation from smaller particles,
and the nucleation mode, which includes
particles smaller than 0.1 μm originating
from combustion processes (e.g. vehicle
exhausts, biomass burning). Natural
aerosol, originating from the sea and the
soil, is mostly in the coarse mode and
is generally considered as less harmful
than anthropogenic aerosol, generated by
combustion sources, found mainly in the
fine mode and able to penetrate deeply
in the respiratory tree. The harmful role
of nanoparticles, able to reach the alveoli
and to be directly transported inside the
body cells, is still a matter of debate. The
chemical composition of an atmospheric
particle depends on its source as well as
on its “story” from the time of its emission
or formation to the time when it reaches
the receptor (e.g. the human body). Some
particles are directly released from their
source into the atmosphere (primary PM),
but the characterisation of any emission
source is quite complex, as it generally
changes with time and operative conditions.
In addition, once formed, particles
often undergo chemical and physical
transformations and for this reason what
is measured at the receptor may be also
very different from what is released at the
source. Even more difficult to trace are the
other particles formed in the atmosphere
as a result of chemical reactions between
gaseous compounds or gas-to-particle
conversion; of particular relevance, in this
framework, is the oxidation of biogenically
released VOCs.
Information about PM sources can be
obtained by analysing their chemical
composition. Only a limited number of
compounds constitute more than 1% of the
overall PM mass: a few metals (Al, Si, Fe), the
main anions and cations (chloride, nitrate,
sulphate, carbonate, sodium, ammonium,
potassium, magnesium and calcium),
elemental carbon and organic material.
This last category is the most important,
as it generally represents 20 – 60% of PM,
CNR Environment and Health Inter-departmental Project
but, unlike the other main components,
it is not a single chemical compound but
it is constituted by many hundreds of
compounds, none of which constitute more
than 1% of the total PM mass. Although
the determination of the listed compounds
is in most cases sufficient to obtain the
mass closure (i.e the sum of the single
components equals the gravimetric mass),
the determination of micro-components is
generally necessary to obtain a picture of
PM sources and effects (35-45). Although
quite complex, the determination of most
inorganic PM components has been one of
the targets of field research during the last
10-20 years. By determining inorganic PM
macro-components it is possible to trace
natural sources (sea-spray, desert dust,
local crustal components) and to measure
the contribution of secondary compounds
(ammonium sulphate and ammonium
nitrate); inorganic micro-components, on
the contrary, may be of help in determining
the contribution of anthropogenic sources,
e.g. dust re-suspension and industrial
sources (46-48). Much less understood
and quantified is the organic fraction, as
the chemical analysis is generally able
to identify no more than 15-20% of total
organic mass.
Once we are able to measure PM
concentration and to determine its chemical
composition, we need to clarify the link
between concentration and exposure. This
is a critical point in the scientific studies
about health effects, as the reference PM
values are generally those measured by
local Protection Agencies, i.e. outdoor
values sometimes taken at traffic hotspots,
while people generally spend most of their
time in indoor environments, including
homes, working places and vehicles, and
only a small part of their time outdoors.
Considerable work is still needed to develop
models able to simulate the behaviour of
individuals in indoor microenvironments.
Also, we need information about the
composition of indoor PM, that may
greatly differ from the composition of
outdoor particles (49-56). For example, in
indoor environments we may be exposed to
much more particles produced by peculiar
sources such as domestic wood burning or
cooking than to particles emitted by traffic
As a consequence of the many difficulties
arising when relating PM concentration
to the results of epidemiological studies,
the scientific community is now trying to
find a relationship between health effects
and individual chemical components.
This attempt requires the availability
of long time series of PM composition
study and is still in its infancy. The other
pathway to elucidate the link between PM
and health effects is the study of the PM
toxicological effects, that is the specific
mechanisms that lead PM to cause the
observed health impacts. These studies
include animal models, human exposure
during occupational activities and
experimental exposures. The mechanisms
of PM effects on human health are still
quite uncertain, but given the variety and
degree of observed health associations, it
is likely that more than one of them are
involved. Basically, particles entering
the tissue cells may cause inflammation;
researchers increasingly find that reactive
oxygen compounds (in the PM or produced
by stimulated cells) play a role. Because of
the presence of particles in the respiratory
tract, changes in the respiratory function
may occur. Particles in the blood may
increase viscosity, causing thrombosis or
myocardial infarct. Of course, individuals
with pre-existing deficiencies of the
cardiovascular or respiratory system may
suffer more severe effects (56-61).
It is clear that a more integrated approach
Role of Atmospheric Pollution on Harmful Health Effects
is needed to get insights into PM health
effects. In particular, the link between
health effect and PM component and size
and the biological specific mechanism
of its action requires further combined
interdisciplinary studies.
3.2.2 Transition (heavy) metals
In contrast to gaseous specific compounds
such as benzene or carbon monoxide,
the assessment of metal and metalloid
compounds in ambient air is complicated
by the fact that different species with
considerably differing toxicity and/or
carcinogenic potency may be encountered.
Therefore, to fully evaluate the health
effects, it is important to know which
compounds do occur in the environment
or at least which compounds form the
main constituents. In ambient air, metals,
metalloids and their compounds are
mainly encountered as part of particulate
matter. They may be present in the non
soluble, non stoichiometric mixture phase
(for example as spinels) or as soluble ionic
compounds (salts). To a lesser extent and
under certain environmental conditions,
gaseous forms (i.e, organometallic
compounds) may or may not be adsorbed
by particles. In respect to their effects on
the environment and on human health,
these compounds can be characterized by
other parameters, such as water solubility
(extended to solubility in biological fluids),
particle size distribution, morphology
and specific surface area, and chemical
heterogeneity of their particles (for
example, a metal compound encapsulated
in another aerosol or surface enrichment of
volatile compounds), or the concentration
of metals and metalloids in the particles
ultimately contacting target tissues in the
human body. All parameters mentioned
will influence bioavailability and possible
effects. In addition, metal and metalloid
containing substances can undergo various
chemical and physical transformations
in the atmosphere on their way from the
source to a possible receptor. For example,
As (III) compounds may be oxidized to
As (V). Unfortunately, analytical methods
normally identify only the elements present
in atmospheric particles, since a specific
analysis is extremely difficult in the
concentration range occurring in ambient
air (typically several ng/m³). In addition,
the state of oxidation may change during
sampling. Consequently, information on
the concentration of different compounds
in ambient air is very limited at present.
Another possibility to gain some insight
into them is to analyze which compounds
are emitted by the most important
natural (i.e, weathering processes) and
anthropogenic sources.
Some metals are naturally found in the
body and are essential to human health.
Iron, for example, prevents anaemia, and
zinc is a cofactor in over 100 enzyme
reactions. They normally occur at low
concentrations and are known as trace
metals. In high doses, they may be toxic to
the body or produce deficiencies in other
trace metals; for example, high levels
of zinc can result in copper deficiency,
another metal required by the body. Heavy
metals (HMs) (or toxic metals) are trace
metals with a density at least five times that
of water. As such, they are stable elements
(meaning they cannot be metabolized by
the body) and bio-accumulative (passed
up the food chain to humans). These
include: mercury (Hg), nickel (Ni),
lead (Pb), arsenic (As), cadmium (Cd),
aluminium (Al), platinum (Pt), and copper
(Cu) (the metallic form versus the ionic
form required by the body). Heavy metals
have no function in the body and can
be highly toxic. Once liberated into the
environment through air, drinking water,
CNR Environment and Health Inter-departmental Project
food, or countless human-made chemicals
and products, heavy metals are taken into
the body via inhalation, ingestion, and
skin absorption. If heavy metals enter and
accumulate in body tissues faster than the
body’s detoxification pathways can dispose
of them, a gradual build-up of these toxins
will occur. High concentration exposure is
not necessary to produce a state of toxicity
in the body, as heavy metals accumulate in
body tissues and, over time, can reach toxic
concentration levels. Human exposure to
heavy metals has risen dramatically in the
last 50 years as a result of an exponential
increase in the use of heavy metals in
industrial processes and products. Today,
chronic exposure comes from mercuryamalgam dental fillings, lead in paint and
tap water, chemical residues in processed
foods, and “personal care” products
(cosmetics, shampoo and other hair
products, mouthwash, toothpaste, soap).
The effects of Heavy Metal toxicity studies
confirm that heavy metals can directly
influence behaviour by impairing mental
and neurological functions, influencing
neurotransmitter production and utilization,
and altering numerous metabolic body
processes. Systems in which toxic
metal elements can induce impairment
and dysfunction include the blood and
cardiovascular system, detoxification
pathways (colon, liver, kidneys, skin),
endocrine (hormonal)system, energy
gastrointestinal, immune, nervous (central
and peripheral), reproductive, and urinary
systems. Breathing heavy metal particles,
even at levels well below those considered
nontoxic, can have serious health effects.
Virtually all aspects of animal and
human immune system functions are
compromised by the inhalation of heavy
metal particulates. In addition, toxic metals
can increase allergic reactions, cause
genetic mutation, compete with “good”
trace metals for biochemical bond sites,
and act as antibiotics, killing both harmful
and beneficial bacteria. For the most toxic
HMs, atmospheric concentrations for Pb,
As, Cd, Ni and Hg in ambient air have
been regulated by European Commission
directives (Directive 1999/30/EC from 22
April 1999 for Pb; Directive 2004/107/EC
from 15 December 2004 for As, Cd, Ni
and Hg). For Pb, an annual limit value of
0.5 g m-3, entered into force 1.1.2005, has
been set. The pertaining As, Cd and Ni
Target Values are, otherwise, reported in
Table 3.1.
Table 3.1 Target values for As, Cd and Ni.
Target Value (1)
6 ng m-3
5 ng m-3
20 ng m-3
For the total content in the PM10 fraction averaged
over a calendar year
Specifically, from chronic arsenic
exposure, the greatest dangers are lung
and skin cancers and gradual poisoning,
most frequently derived from living near
metal smelting plants or arsenic factories.
Arsenic toxicity has been recognized
for centuries, and hair shows significant
correlation with its intake. As can be
released to the atmosphere from metal
transformation, fuel combustion and the
use of pesticides.
In the air, As exists predominantly absorbed
on particles, and is usually present as a
mixture of arsenate (As(+V)) and arsenite
(As(+III)), except in areas of arsenic
pesticide application or biotic activities,
where organic species are predominant
(27,62). Recent data display a wide range
of As concentrations in atmospheric
particulate matter, for samples collected
Role of Atmospheric Pollution on Harmful Health Effects
at various sites in Spain (ranging from
13 to 144 mg kg-1 for a rural area and an
industrialised area, respectively) (63). All
studies performed in the Mediterranean
basin (64-65) agree with an enrichment of
As in the atmosphere as shown by (66).
Cadmium is an element that is naturally
found in the earth’s crust. Cadmium is often
found as part of small particles present in
air. Cadmium has many uses in industry
and consumer products, mainly batteries,
pigments, metal coatings, and plastics.
Cadmium can enter the environment in
several ways. It can enter the air from
the burning of coal and household waste,
and metal mining and refining processes.
Cadmium attached to small particles may
get into the air and travel a long way before
coming down to earth as dust or in rain or
snow. Cadmium does not break down in the
environment but can change into different
forms. Most cadmium stays where it enters
the environment for a long time. Cadmium
has no known good effects on health.
Breathing air with very high levels of
cadmium severely damages the lungs and
can cause death. Breathing lower levels
for years leads to a build-up of cadmium
in the kidneys that can cause kidney
disease. Other effects that may occur after
breathing cadmium for a long time are
lung damage and fragile bones. Workers
who inhale cadmium for a long time may
have an increased chance of getting lung
cancer. The greatest danger from chronic
nickel exposure is lung, nasal, or larynx
cancers, and gradual poisoning from
accidental or chronic low-level exposure,
the risk of which is greatest for those living
near metal smelting plants, solid waste
incinerators, or old nickel refineries. Nickel
combined with other elements is naturally
found in the earth’s crust, in all soils, and
it is also released from volcanoes. Nickel
is the 24th most abundant element, and in
the environment it is found primarily in
the form of oxides or sulphides. Nickel
is also found in meteorites and in lumps
of minerals on the bottom of the ocean,
and it is known as sea floor nodules. The
earth’s core is believed to contain large
amounts of nickel. Nickel is released into
the atmosphere during nickel mining and
by industries that convert scrap or new
nickel into alloys or nickel compounds
or by industries that use nickel and its
compounds. It is also released into the
atmosphere by oil-burning power plants,
coal-burning power plants, and trash
incinerators. The nickel that comes out
power plants’ stacks is attached to small
particles of dust that settle to the ground
or are transported in the air by rain. It
will usually take many days for nickel to
be removed from the air. If the nickel is
attached to very small particles, removal
can take longer than a month. Given nickel’s
ability to cause contact dermatitis, and its
observed perturbation of immunoglobulin
levels, elevated hair levels may serve as an
indicator of possible immune dysfunction,
as well as a potentially useful marker of
cardiovascular problems.
Pb is one of the most released HMs in the
Mediterranean atmosphere. Traffic remains
the main source at global scale (67), but its
relative importance varies from region to
region. In Europe, the phasing out of alkylleads in gasoline resulted in a decrease
in atmospheric lead concentrations. As a
result, industrial emissions (lead smelting
and steelworks) became predominant
in Europe and discernable from traffic
emissions at continental scale (68). The
annual emission of Pb in the Mediterranean
region has been estimated to be about
1.1 104 Mg yr-1 in 2005 by (69). Lead is
a known neurotoxin (it kills brain cells),
and excessive blood lead levels in children
have been linked to learning disabilities,
CNR Environment and Health Inter-departmental Project
attention deficit disorder, hyperactivity
syndromes, and reduced intelligence and
school achievement scores.
3.2.3. Mercury (Hg) and impact on human
Although mercury is an element found in
nature and as such it will always be present
in the environment, human activities have
significantly increased global atmospheric
mercury deposition since pre-industrial
times. A significant increase in mercury
emissions in the atmosphere occurred
during the industrial revolution due to fossil
fuel combustion and other human activities.
Mercury is today a severe and chronic
pollution problem in the environment.
It is released into the atmosphere from
a variety of anthropogenic (i.e, cement
production, waste incineration, power
generation facilities, smelters) (70-71) and
natural sources (i.e, volcanoes, crustal
and vegetation degassing, oceans) (7274) in different chemical and physical
forms.(75). In the troposphere, the most
important forms are gaseous elemental
mercury (Hg0), divalent reactive gaseous
mercury, Hg(II), which consists of various
oxidised compounds, and particle-bound
Hg (Hg-p), which consists of various
Hg compounds. It should be noted that
information on the speciation/fractionation
of these different chemical and physical
forms is largely operationally defined.
Conversions between these different
forms provide the basis of Hg’s complex
distribution pattern on local, regional
and global scale. Hg cycles in different
environmental compartments depends
on the rate of different chemical and
physical mechanisms (i.e, dry deposition,
wet scavenging) and meteorological
conditions as well as on the anthropogenic
variables that affect its fate in the global
environment. Experimental field data and
model estimates indicate that anthropogenic
mercury emissions are at least as great as
those from natural sources, and contribute
to the global atmospheric pool. A threefold
increase of mercury deposition since preindustrial times was in fact observed from
the analysis of lake sediments, ice cores
and peat deposits in both hemispheres (7479). Recent studies have highlighted that
in fast developing countries (i.e, China,
India) mercury emissions are rapidly
and dramatically increasing due mainly
to a sharp increase in energy production
from coal combustion. Recent emission
estimates highlighted that the Asian
emissions are considered to have a global
Evidence shows that an increase in
ambient air levels of mercury is linked
to an increased load of toxic mercury in
ecosystems (80). The atmospheric input
of this element in aquatic and terrestrial
ecosystems is driven by particle dry
deposition and wet scavenging by
precipitation mechanisms (81-83). The
most important from a toxicological point
of view are the metallic forms. In fact,
the impact of mercury on human health
and the environment depends on several
mechanisms, which, in turn, depend on
the toxicokinetic of its major chemical
forms present in different environmental
media including elemental mercury
(Hg0), inorganic mercury (i.e, HgCl2) and
organic mercury (i.e, methylmercury).
This toxicokinetic mechanisms include
absorption, distribution, metabolism and
excretion. Therefore, according to the
relevant chemical form of mercury, the
combination of these mechanisms will
determine the risk associated to the human
exposure. For instance, the absorption of
Hg0 vapour occurs rapidly through the
lungs, but it is poorly absorbed from the
gastrointestinal tract.
Role of Atmospheric Pollution on Harmful Health Effects
Once absorbed, elemental mercury is
readily distributed throughout the body,
it crosses both placental and blood-brain
barriers (84-86) Elemental mercury is
oxidised to inorganic divalent mercury by
the hydrogen peroxidase-catalase pathway,
present in most tissues. The distribution
of absorbed elemental mercury is limited
by the oxidation of elemental mercury
into mercuric ion as the mercuric ion has
a limited ability to cross the placental
and blood-brain barriers. Once elemental
mercury crosses these barriers and is
oxidised to mercuric ion, its return to
the general circulation is impeded, and
mercury can be retained in brain tissue.
The elimination of elemental mercury
occurs via urine, faeces, exhaled air, sweat
and saliva. The excretion pattern depends
on the extent to which elemental mercury
has been oxidised to mercuric mercury
(87-90). Absorption of inorganic mercury
through the gastrointestinal tract varies
with the particular mercuric salt involved
and decreases with its increasing solubility
and can reach even 20% (91). Available
data indicate that absorption of mercuric
chloride from the gastrointestinal tract
results from an electrostatic interaction
with the brush border membrane and
limited passive diffusion. Increases in
intestinal pH, high doses of mercuric
chloride causing a corrosive action, a milk
diet and increases in pinocytotic activity
in the gastrointestinal tract have all been
associated with increased absorption of
inorganic mercury. Inorganic mercury has
a limited capacity of penetrating bloodbrain and placental barriers. There is some
evidence indicating that mercuric mercury
in the body following oral exposure can be
reduced to elemental mercury and excreted
via exhaled air. Because of the relatively
poor absorption of orally administered
inorganic mercury, most ingested doses in
humans are excreted through the faeces.
Methylmercury is rapidly and extensively
absorbed through the gastrointestinal tract
(92). Absorption information following
inhalation exposure is limited. Epidemic
of mercury poisoning following high-dose
exposures to methylmercury in Japan and
Iraq demonstrated that neurotoxicity is
the most worrisome health effect when
methylmercury exposure occurs to the
developing foetus. Dietary methylmercury
is almost completely absorbed into
the blood and distributed to all tissues
including the brain. It also readily passes
through the placenta to the foetus and
foetal brain. Methylmercury in the body is
considered to be stable and it is only slowly
demethylated to form mercuric mercury
in rats. It has a relatively long biological
half-life in humans (44-80 days) and it is
excreted through faeces, breast milk and
4.1 Volatile organic compounds
The atmosphere is formed by a restricted
number of macro-components, namely
gaseous nitrogen, oxygen, argon, carbon
dioxide, solid particulate matter and water;
the latter exists as vapour, or in liquid
and solid forms. Thousands of microcomponents are dispersed in the gas phase
and/or participate to aerosol composition
. Although occurring often at very low
levels (even below one part-per-trillion),
they nevertheless intervene in the
physics and chemistry of the atmosphere,
heavily influencing our life. Among the
micro-components, a key role is played
by hydrocarbons and their derivatives,
cumulatively called “volatile organic
compounds” (VOC), which are in gas form.
Congeners of VOC occur in particulates
as adsorbed on soot or dissolved in water
CNR Environment and Health Inter-departmental Project
drops and crystals.
In its chemical structure, VOC include: i)
aliphatic hydrocarbons (linear, branched
and cyclic); ii) arenes having at least one
aromatic group (Ar), namely benzene,
alkylbenzenes and some polyaromatics
(naphthalene); iii) alcohols (ROH) and
ethers (ROR1); iv) carbonyls, comprising
aldehydes (RCHO) and ketones (RCR1O),
v) carboxy-acids (e.g. formic, HCOOH) and
phenols (ArOH); vi) organic halogenides;
vii) nitrogen, sulphur and phosphorus
compounds; ix) heterocyclic and x) mixed
functionality types (94-96).
Widely varying in concentration, structure
and properties, different classifications
have been proposed for VOC. One worth
of mention distinguishes four groups of
substances according to major aftermaths
induced onto the environment, and their
chemical formulas. They are:
i) gases promoting the Earth warming
(greenhouse effect VOC); ii) compounds
responsible for the stratospheric ozone hole;
iii) hydrocarbons promoting (or involved
in) the tropospheric ozone and secondary
particulate generation (photochemical
smog); and iv) toxic compounds. At this
regard, it seems worth to remark that:
- The global Earth warming has been
overall associated with carbon dioxide
and water vapour. Nevertheless, it is well
known that other compounds promote
this phenomenon, e.g. methane, nitrogen
protoxide, sulphur hexafluoride, and
chlorofluorocarbons (freons or CFC)
(97-98). The worldwide use of these
substances and/or their release into the
environment as by-products of human
activities caused a strong increase in
their respective atmospheric loads,
and long, expensive and concerted
actions must be launched to control and
remediate global warming.
- The “ozone hole” first observed on
stratosphere Antarctica has been
associated with CFC, which capture
the solar light and start a reaction chain
with those transforming molecular
oxygen (O2) into ozone (O3). The variety
and intensity of CFC use, combined
with their long lifetime, caused their
accumulation in the air and transport
by winds in remote regions. This
phenomenon leads to an increased ultraviolet radiation that reaches the ground,
causing in particular an increase in
skin tumours. This is the reason why
CFC have been banned and replaced
by other chemicals characterized by
shorter lifetimes; nevertheless, they
have lower heat capacities and higher
prices, so their true use gains place
with difficulty (99-100).
- At ground level, ozone represents a
sanitary risk for humans and causes
damages to crops and materials, thanks
to its strong oxidant potency. Ozone
is primarily generated in reactions
involving oxygen, nitrogen oxide
and dioxide, and “active” sunlight (l
<430 nm) (93). The natural ozone
background regulated and limited by
them. Nevertheless, this equilibrium
is modified by VOC that trigger
processes leading to the formation
of ozone without NO2 consumption.
Thus, the ozone concentration can
largely increase. A lot of other oxygencontaining compounds are generated
in the form of molecules (carbonyls,
carboxy-acids, phenols, epoxides) and
free radicals (OH, RO, RO2, HO2);
when oxidized, hydrocarbons easily
condensate giving raise to nuclei around
which nano-particles are formed. Not
all hydrocarbons participate equally
to the photochemical smog formation.
Methane, short-chain alkanes and
benzene are quite non-reactive,
Role of Atmospheric Pollution on Harmful Health Effects
similarly to acids and ketones. By
contrast, alkenes and alkylbenzene
have high photochemical ozone
formation potentials (101). A special
role is played by isoprene and terpenes
(biogenic hydrocarbons), which are
very reactive vs. OH and NO3 radicals
as well as vs. O3; they can induce high
airborne concentrations of O3 in rural
areas (forests, crops). Through nanoparticle and oxidant formation, reactive
VOC indirectly affect the air quality.
Some VOC have been recognized as
toxicants for their acute and/or chronic
effects and have been included in the
priority list of pollutants (102). For
instance, benzene is known as tumour
promoter, similarly to many halogensubstituted hydrocarbons (e.g. bromoform,
methyl tetrachloride). Numerous VOC are
carcinogenic (butadiene, diazomethane),
mutagenic (chloroform) or induce
cough, skin, eye and noose sensitization,
throat irritation (aldehydes, organic
halogenides), faint, loss of knowledge
(tetrachloroethylene, methanol, xilene),
diarrhoea, liver and kidney damages
(aniline); some of them are poisonous
(ethylene oxide, camphor, methanol,
monomethyl mercury), or psycho-active
inducing euphoria, depression, headache
(methyl bromide, ethanol). Acids, bases
and strong solvents are also caustic
(trichoroacetic acid; dimethylamine;
methanol, acetone, chlorobenzene).
As far as the above mentioned features
of VOC have been clarified, dedicated
legislations have been issued to preserve
the environment and health (103-112).
Regulations refer to industrial and car
emissions, power and heat production,
agriculture, waste management, materials,
food, open air, indoor and work places.
4.2. Organic particulate matter
The organic fraction of particulates is well
known, yet much remains not understood.
Thousands of chemical substances have
been identified in airborne and emission
particles. They include n-alkanes and
non-polar aliphatic or alkylbenzenes,
polynuclear aromatic hydrocarbons (PAH)
and the corresponding alkyl-, nitro-,
amino-, carbonyl-, oxy-, sulphur- and azaderivatives, halogen- and phosphoruscontaining pesticides, phthalate esters,
acids and phenols, alcohols and nitriles
(113-115). Most of these substances
show scarce hydro-affinity, whilst shortchain, mixed functionality acids are
water soluble. Finally, polymers and
macromolecules, often of biogenic origin,
contribute to the bulk (116). Several
studies have been carried out to elucidate
chemical composition, with two main
objectives in mind: to draw an indirect
toxicity parameterization of emissions
or environment, to solve the biogenic/
anthropogenic origin of particulates
and evaluate the relative contribution
of their sources (117-121). Finally, the
detection of reactive compounds and their
corresponding by-products has allowed to
put in evidence the oxidation capacity of
the atmosphere, deriving from both the
presence of oxidants (ozone, free radicals,
nitrogen oxides) and light (122). This
seems particularly important in the case
of reactions involving gaseous substances
that are converted into particles (secondary
pollution), and whenever the degradation
products are much more toxic than their
parent compounds (116). Despite extensive
and prolonged efforts made in this field,
the complete characterization of organic
fraction of particulates is far from being
reached. It has been demonstrated that
the organic fraction accounts for 10 to
over 80% of total airborne particulates.
CNR Environment and Health Inter-departmental Project
This variability widely depends not
only on the environmental contour
investigated (locality, emission sources
impact, orography, meteorology, indoor or
outdoor), but also on the approach adopted
to measure it. In particular, very different
results are found if the sole “organic solvent
extractable fraction” is considered, the
water soluble fraction is taken in account,
or optical automatic methods are adopted.
Otherwise, the WSOC of aerosol contains
many different compounds that, to date,
are poorly characterized. Experimental
evidence suggest that these compounds
are at least partly responsible for the main
oxidizing and toxicant properties of urban
and non-urban aerosols (56,116). Unlike
elemental carbon, which is generally
associated with the presence of humans
(the sole exceptions consist of forest
spontaneous fire and volcano emissions),
the organic fraction of particulates has a
twin origin, i.e. biogenic and anthropogenic.
That can be easily explained through
an example. Linear alkanes globally
account for a few percents of organic
particulate matter. Their group presents
two well distinct composition behaviours.
The saw-tooth distribution dominated
by long-chain odd homologues (C29H60,
C31H64 or C33H68) is typical of biogenic
sources (e.g. high vegetation), whilst the
mono-modal (or bell shaped) distribution
characterizes anthropogenic emission
like motor vehicles; in this case, the
maximum centred at C19H40÷C25H52. With
the exception perhaps of macromolecules
and acid compounds having many
hydrophilic groups (carboxyl-, hydroxyl-,
carbonyl-, epoxy-), organic substances are,
individually, micro-components of soot
accounting for parts per million down to
parts per trillion of the particulate mass.
Together, they form a mixture often
adhering to the surface, while the particle
core is made of elemental carbon and
inorganic elements. The polarity and size
of molecules influence the hydro-affinity
of particles, their growth capacity and
then their time life in the atmosphere.
Most organic compounds are neutral,
although exhibiting different polarities;
basic species are a few (they include
aromatic amines and aza-PAH), whilst
a lot are acidic (phenols, carboxy-acids),
even when lypophilic. This variety is of
environmental concern, since its mediumand long-term toxicity is strictly dependent
on the polarity features of these elements.
Studies conducted in Italy and abroad have
demonstrated that organic fraction is a
major contributor to particulate toxicity. In
Table 5.2. UFPs/NPs natural and
anthropogenic sources.
Intentional (NPs)
Controlled size and
shape, designed for
Forest fires
Power plants
metal oxides,
carbon, polymers
(hot lava)
-wires, -needles,
-tubes, -shells,
-rings, -platelets
Jet engines
Untreated, coated
applied to many
products: cosmetics,
medical, fabrics,
electronics, optics,
displays, etc.)
Metal fumes
welding, etc.)
Polymer fumes
Other fumes
Heated surfaces
Electric motors
Role of Atmospheric Pollution on Harmful Health Effects
fact, up to 90% of total carcinogenic and/or
mutagenic potency of particulates results
to be associated to the corresponding
organic extracts, although the substrate
can act as promoter of synergic effects.
Two main aftermaths can be detected, the
former acute (cyto-toxicity, causing cell
death) and the latter chronic (cell damage,
inducing carcinogenicity, mutagenicity,
teratogenicity). Both of them generally
appear as associated to neutral polar and
acidic compounds taken as pools, and
a number of very strong toxicants have
been identified among them, including
nitro-lactones and dioxins (123-124). By
contrast, non-polar components are not
toxic, although can exalt the potencies
of active species. Despite that, the true
contribution provided by each primary
toxicant to environmental toxicity cannot
be evaluated, since the interaction with the
matrix is neglected in terms of synergic/
antagonistic actions, and with respect to
net exposition of humans. At this regard,
two points must be taken in account.
First, due to combination of its nature
and airborne concentration, to ambient
conditions (temperature and humidity),
to soot concentration and characteristics,
each organic species partitions between
gas and particulate (125). Thus, their
interaction with human body is variable.
Secondly, the total load of organic
compounds is distributed among the
particle size fractions, with the general
tendency to accumulate into the ultra-fine
and fine particles (126-127). It is worth
remarking that the strongest toxicants (e.g.
PAH, chlorinated hydrocarbons, nitrocompounds) accumulate into the ultrafine and fine particles, while components
with less pronounced potencies are
more equally distributed. Very polar
components, deriving from the oxidation
of gaseous emissions, act as condensation
nuclei, triggering the formation of nanoparticles. These latter are considered, as
a consequence of their number and size,
the main cause of air toxicity and an
emerging “hot issue” for environmental
safety preservation. Thus, the association
of organic species to them has heavy
environmental consequences.
The importance of the organic component
of particulates is well depicted by
legislation issued to preserve health, and
especially the health of workers (103106, 128-130). In fact, a set of organics is
listed by WHO (UNEP) among the most
important toxicants; these include a dozen
of persistent pesticides, polychlorinated
dioxins, furans and biphenyls. On the
other hand, PAH are quoted in European
Union Directives concerning air quality or
integrated pollution prevention and control.
The target value of 1 nanogram per cubic
metre of air, calculated as yearly average,
has been established for benzo(a)pyrene,
kept as an index of aerosol carcinogenicity.
European and Italian legislations require
to measure also other seven relevant
carcinogenic PAH. Toxicants like dioxins
and furans are also quoted somewhere,
however non target or limit values are
In terms of future actions, certainly the role
played by organics in particle generation
and accumulation, in troposphere
properties (e.g. radiance, heat absorption,
albedo, water condensation, global
warming), human health and environmental
preservation has a key importance. Concern
would be ascribed also to “new” pollutants
like polyfluorinated acids, pesticides and
plastic surrogates, psychotropic substances
and poly-functionalized species (oxyacids,
polycarboxylic acids, which are suspected
to affect our world.
CNR Environment and Health Inter-departmental Project
4.3. Research in organic air pollutants:
the past, the present and the future
Organic substances are at the core of a
series of studies conducted by CNR with
the aim of clarifying the features and
dynamics of the environment, in the frame
of international and national programmes.
Special attention has been paid to the
composition of the atmosphere, the effects
induced by anthropogenic activities and
by natural events and sources, the kinetics
of pollutants in the presence of oxidants
and light, the assessment of mobile and
stationary emissions and pollution sources,
particulate generation and properties, and
sanitary relevance of pollution. According
to recent developments, some items seem
to deserve better analysis: the clean (green)
energy generation, the chemistry of radicals,
the air/sea, air/ice and air/soil interactions,
the influence on meteorology and climate,
the strict relationships between chemical
composition and toxicity. As concerns
this latter, interesting issues have been
identified in nanoparticles, water-soluble
organics, organo-metallic compounds
and psychotropic substances. Finally, the
economic, social and legislative aspects
cannot be neglected in view of life quality
5.1 Sources and formation of ultrafine
Ultrafine particles are the dominant
contributors of particle numbers in the
PM2.5 fraction. Particles with dimensions
< 100 nm are defined as nano-sized and
ultrafine particles (NSPs, UFPs), according
to their manufactured or environmental /
biological origin. In particular, NSPs are
called ‘ultrafine’ particles (UFPs) also
by toxicologists, ‘Aitken and nucleation
mode’ particles by atmospheric scientists
and ‘engineered nanostructured materials’
by materials scientists(132). The simplest
examples of naturally occurring ultrafine
particles are those found in the biological
tissues of organisms. For example, biogenic
magnetite is a natural NSP found in many
animal species. Other nanosized materials,
including fullerenes, are naturally
originated from combustion processes
such as forest fires and volcanoes. Sources
of ambient UFPs are either primary or
secondary. In urban environments, the
dominant contributors of primary UFPs
are combustion products found in motor
vehicle exhausts, which are usually
black carbonaceous soots with particle
dimensions 200–300 nm. Atmospheric
oxidation of gas-phase primary exhaust
species can produce lower vapor pressure
compounds that readily condense onto
existing particles and produce secondary
mass. Particle composition and size can
also evolve due to interaction/reaction of
gas- and liquid- or solid-phase species
at the particle surface or in the bulk
solution, as well as through coagulation
of existing particles. In some conditions,
photochemical oxidation of gas-phase
species can directly produce new UFP
(133). Most atmospheric UFPs are usually
<50 nm and evidence has been found
that the particle count distribution peaks
at 20–30 nm at roadsides with heavy
traffic134. Diesel-exhaust particles (DEP)
are major components of PM2.5. Although
installing a diesel particle filter (DPF) can
reduce the number of larger particles in
the exhaust, nanoparticles are produced
during DFP regeneration. Environmental
or atmospheric UFPs contain semivolatile
alkanes that originate from fuels and
lubricants (135). In particular, primary
exhaust emissions of particles consist
Role of Atmospheric Pollution on Harmful Health Effects
on the type of product. In table 5.1 (136)
main differences between environmental
and manufactured UFPs are reported.
≤50 nm (≤100 nm:
1–100 nm (biomedical
nanoparticles can be larger
than 100 nm)
Two or three
dimensions are on
the nanoscale
Three dimensions are
on the nanoscale (nanoobjects with one and two
nanoscale dimensions
are called nanoplates and
nanofibers, respectively)
Table 5.1. Differences between
environmental and manufactured
Nano- : particles, spheres,
tubes, rods, fibers, wires,
ropes, sheets, eggs,
liposomes, dendrimers,
mainly in a mixture of elemental carbon
and organic compounds, and traces of
heavy metals and sulphur. Tire wear also
contains carbonaceous materials, while
brake wear is rich in heavy metals136.
Moreover, particle hygroscopic growth
measurement techniques have been used
to show that ultrafine particles may show
two separate modes of ‘‘less’’ and ‘‘more’’
hygroscopic particles (137). The less
hygroscopic particles are those that exhibit
little or no growth when exposed to a high
relative humidity (RH) (typically 80–90%)
and are thought to be mainly composed
of hydrophobic chemicals such as waterinsoluble organic compounds and soot, and
must be attributed to primary emissions
from traffic and other combustion sources.
More hygroscopic particles grow by a
larger factor (e.g, 1.38–1.69 for 35–265
nm particles at 90% RH) and have been
shown to contain inorganic chemicals such
as nitrate, sulphate, sodium and potassium
and sometimes organic carbon as well. The
more hygroscopic particles can result from
the conversion of gaseous compounds into
particles, or from the modification and
oxidation of pre-existing particles. Such
chemical and physico-chemical processes,
named “secondary”, may occur even
far from the emission sources, and may
influence the concentrations of ultrafine
and fine particles outside urban areas. It has
been shown that the nucleation of ultrafine
particles from condensation of reactive
gases is responsible for the increase of
ultrafine particle number concentrations in
many rural areas, including polluted and
natural sites. The hygroscopic behaviour
of the aerosol is an important quality, as
this will determine how they interact with
clouds (138), which, in turn, affects the
lifetime of the particles in the atmosphere.
Conversely, components of manufactured or
engineered nanoparticles vary, depending
Carbon soot,
(alkanes), heavy
metals, sulfur
Carbon (fullerene,
nanotube), metal oxide
(TiO2, ZnO), CdSe,
metalloids, transition
metals, polymers
More recently, even smaller particles in the
nucleation mode with peak diameters around
4 nm have been observed. Humans have
been exposed to airborne NSPs throughout
all their evolutionary stages. Nevertheless,
such exposure has increased dramatically
over the last century due to anthropogenic
sources. These can be classified as
unintentionally or intentionally produced,
depending on weather UFPs represent a
sub-product or the major product coming
from an anthropogenic source. Although
the mass of UFPs in ambient air is very
low, approaching only 0.5–2.0 μg/m3 at
background levels, it can increase severalfold during high pollution episodes or on
highway. In urban areas motor vehicle
particle emissions are a dominant pollution
source, where more than 80% of particle
number concentrations are found in the
CNR Environment and Health Inter-departmental Project
ultrafine size range. However, very little
information can be obtained about particle
number from particle mass measurements,
and as current air quality standards are
mass and not particle number-based, this
means that the greater proportion of motor
vehicle particle number emissions are not
controlled or regulated (141). The latter
case is represented by nanotechnologies.
Main natural, unintentional and intentional
sources of NSPs are reported in table 5.2
5.2 Role of UFPs and nano-particles as
atmospheric toxicants
Inhalation of particulate matter leads to
pulmonary inflammation and reduction in
lung function (142) with secondary systemic
effects or, after translocation from the lung
into the circulation, to direct toxic effects
on cardiovascular function (143) and on the
coagulation pathway thus contributing to
the onset of coronary events (144). Through
the induction of cellular oxidative stress and
proinflammatory pathways (144), particulate
matter augments the development and
progression of atherosclerosis (145).
The main factor of these adverse health
effects seems to be combustion-derived
nanoparticles that incorporate reactive
organic and transition metal components.
An important source of these particles is
new diesel cars with oxidizing converters,
such as modern taxis in North Europe.
Many epidemiological, human clinical,
and animal studies showed that ultrafine
particles (UFPs) penetrate deeply into the
lungs initiating an inflammatory response
leading to respiratory diseases and may
be absorbed directly into the circulating
blood, causing cardiovascular diseases146.
Recent studies highlighted the importance
of identification of susceptible subpopulations and mechanisms of involved
effects. Several chronic clinical conditions
are good candidates to define the susceptible
population to the acute effects of UFP,
while elevated levels of oxidatively altered
biomolecules are important intermediate
endpoints that may be useful markers in
hazard characterization of particulates.
Overall, despite the increasing amount
of data provided by both laboratory and
field studies, the nature of the fraction
of aerosol particles responsible for
health effects is still a matter of debate.
The issue is of importance, because
the different constituents of the aerosol
exhibit distinct sources and emission/
formation processes (147-148). Therefore,
linking toxicological and epidemiological
impacts of atmospheric particulate matter
to chemical composition is a key for the
evaluation of effective pollution abatement
strategies (149-150).
The potential role of UFPs as strong
toxicant species of ambient air derives also
from the following considerations (151):
1. smaller particles have a greater total
surface area per unit of mass than
larger particles; thus, for a given mass,
smaller particles may present a larger
surface area for interacting with airway
tissue or for transporting toxic material
associated with the particle surface into
the airways.
2. In vitro studies suggest that ultrafine
particles may not be as effectively
phagocytosed (ie, ingested for removal)
as larger particles by cells of the innate
immune response.
3. On the basis of size, models predict
that a higher proportion of ultrafine
particles of ~ 20 nm than of larger
particles reach the air-exchanging
alveolar region of the lung. On the basis
of mass, however, more larger particles
than smaller particles reach this lung
4. When particles have been instilled intra-
Role of Atmospheric Pollution on Harmful Health Effects
tracheally into animals, on the basis
of mass, ultrafine particles were more
effective than fine particles in inducing
airway inflammatory responses.
5. In some recent studies ultrafine particles
appeared to move rapidly out of the
airways and into the circulation.
Similarly to gases, when inhaled, specific
sizes of UFPs are efficiently carried by
diffusional mechanisms in all regions of
the respiratory tract. The greater surface
area per mass compared with larger-sized
particles of the same chemistry renders
NSPs more active biologically. This activity
includes a potential for inflammatory and
pro-oxidant, but also antioxidant, activity,
which can explain early findings showing
mixed results in terms of toxicity of
NSPs to environmentally relevant species
(150). It has not been well investigated
whether nanoparticles are responsible for
pulmonary and extrapulmonary health
effects of PM2.5, although the fine particles
are reportedly associated with mortality
from cardiovascular diseases (149).
Nanoparticles can permeate through tissue
walls, translocate to other tissues from the
deposition sites, and cause cardiovascular
dysfunction. However, we do not have a
clear answer as to how far nanoparticles
have a more distinctive toxicological
aspect and are more toxic than larger
particles (151).
concentrations of NSPs per given mass
will likely be of toxicant significance when
these particles interact with cells and subcell components. Likewise, their increased
surface area per unit mass can be of key
importance. The small size facilitates
uptake into cells and transcytosis across
epithelial and endothelial cells into the blood
and lymph circulation, to reach potentially
sensitive target sites such as bone marrow,
lymph nodes, spleen, and heart. Access to
the central nervous system and ganglia via
translocation along axons and dendrites
of neurons has also been observed. NSPs
penetrating the skin distribute via uptake
into lymphatic channels. Endocytosis and
biokinetics are largely dependent on NSP
surface chemistry (coating) and in vivo
surface modifications (151).
The importance of surface area becomes
evident when considering that surface
atoms or molecules play a dominant role
in determining bulk properties; the ratio
of surface to total atoms or molecules
increases exponentially with decreasing
particle size, as reported in table 4.3
Table 5.3. Particle number and particle
surface area per 10 μg/m3 airborne
Particle no.
surface area
There are many debates about the dosemetric which best describes the toxicity
of manufactured nanoparticles. The most
commonly accepted dose metric is probably
the surface area. Particle shape (e.g. fibrous
or spherical), chemical composition, and
the chemistry of the particle surface,
including the zeta-potential, are also
important factors that determine the
toxicity of nanoparticles. It has been
reported that the carcinogenic potency and
toxicity of asbestos (151) largely depend
on fiber length. Fibrous titanium dioxide
particles have been shown to be much more
cytotoxic than spherical nanosize titanium
dioxide particles to alveolar macrophages
(149). Special attention should be paid to
CNR Environment and Health Inter-departmental Project
fibrous nanoparticles, because fiber length
may be predominant metric determining
the toxicity of biopersistent fibrous
5.3 Innovative techniques of chemical–
physical characterization of the UFP
A major limitation of traditional impaction
and membrane technologies is that the
detection limits of the laboratory analysis
instrumentation necessitate the collection
of a large enough mass of sample for
analysis, so temporal resolution is limited
to greater than several hours for ambient
samples. Also, the sample may be affected
by evaporation or condensation of semivolatile components during or after
sample collection and chemical reactions
may take place within the sample itself
or between oxidants in the sample gas
and the collected particles, affecting
the results (148). The size resolution of
these instruments is generally poor to
moderate because of the limitations of
aerodynamic particle separation and
the need to increase sampling times and
analyze a larger number of samples with
increasing size resolution. Nevertheless,
examples exist of nano-size impactors
(e.g. NanoMoudi by MSP Corporation,
USA) and thermophoretic precipitators
(152) which allow single particle collection
aimed at analysis of UFPs chemical
composition, i.e. by scanning transmission
electron microscopy (STEM).
As a matter of fact, anyway, the size
distribution of ultrafine particles is
expected to evolve rapidly in urban air
and such knowledge is essential to the
evaluation of human exposure. Differential
mobility analyzers (DMA) and other
particle evolution / growth measurement
techniques have been used since recent
years to evaluate the behaviour of airborne
UFPs in the ambient air, especially at
urban sites. In example (153), combined
the use of the scanning mobility particle
spectrometer (SMPS), electrical low
pressure impactor (ELPI) and TEM
techniques to characterize the evolution
of the particle size, morphology and
composition distribution during dispersion
of traffic – related emissions at Birmingham
(UK). The ELPI (Dekati, Finland)
measures real-time particle concentration
and size distribution and the SMPS (TSI,
USA) measures particle size distributions
with a high resolution. Combined use of
these two instruments enhances the quality
of particle size distribution measurements.
Measurement size range of the ELPI is 3010000 nm, 50% cut size and the SMPS is
9.6-352 nm. The analytical problem posed
by the wide range of atmospheric PM sizes
is dynamic range. A 1 nm diameter particle
might have a mass of 10-21 g (zeptograms),
whereas a 10 mm particle a mass of 10 ng.
Reliable sampling and accurate chemical
composition determination of a single
nanogram particle is a tough analytical
challenge; for a zeptogram particle, it is
nearly impossible. In addition to PM size,
composition and mass loading, many
other physical and chemical properties
are of interest, and should be preferably
measured simultaneously. Research on
climate changes, i.e, needs to correlate size
and chemical composition with a particle’s
ability to scatter and absorb radiation from
the infrared to the near ultraviolet; health
scientists hypothesize that a particle’s
surface area and surface composition may
be a key to understanding how it interacts
with lung tissue to affect pulmonary
functions and transfer chemicals into the
blood stream (152); finally, the physical
phase (liquid or solid), surface area, and
surface composition can strongly affect
the interaction of atmospheric trace gases
with airborne particles, impacting the
chemical composition of both the gaseous
Role of Atmospheric Pollution on Harmful Health Effects
and condensed phase components of the
Real-time instruments that measure
physical properties such as particle number
densities, mass loadings, and particle
mobility or aerodynamic size distributions
have been available since recent years.
However, real-time instruments that
characterize the chemical composition of
atmospheric PM, ideally as a function of
particle size, are a more recent development
(153). Some near real-time PM chemical
composition instruments, operating with
measurement cycles of 10–60 min, can
characterize the average PM content of
one or more key PM constituents for
an ensemble of particles in a size range
defined by the sample collection system.
Examples include the particle into-liquid
sampler (PILS) that utilizes automated
ion chromatography to quantify average
major anion or cation PM content or an
automated carbon analyzer to determine
the water-soluble organic carbon (OC),
instruments based on particle collection
followed by thermal decomposition and gas
phase chemiluminescence or absorption
spectroscopy allow for semi-continuous
measurements of sulfate and nitrate (153
and references therein). However, the
universal nature of mass spectrometric
detection for atomic and molecular
species makes this technique eligible
as most comprehensive and sensitive
to characterize the chemical content of
atmospheric PM. Over the past decade,
several research groups have made major
strides in adapting mass spectrometric
techniques to meet this challenge and
three major directions of evolution of mass
detection techniques can be currently
identified. One major theme involves the
use of lasers to both vaporize and ionize
individual atmospheric particles sampled
into a mass spectrometer’s source region.
This class of instruments focuses on single
particle measurements. A second class of
aerosol mass instruments uses thermal
vaporization of individual or collected
particles followed by various ionization
techniques. Separation of the vaporization
and ionization steps enables quantitative
detection of PM chemical composition and
mass loading. In addition, the simplicity
of thermal vaporization allows the use
of a variety of ionization techniques that
will produce less sample fragmentation
than traditional electron impact (EI)
methods, such as chemical ionization
techniques(154). However, the most widely
used technique is thermal vaporization
aerosol mass spectrometer (AMS), which
was designed and developed at Aerodyne
Research, Inc. (ARI). The initial version
of the ARI AMS was designed to measure
the real-time non-refractory (NR)
chemical speciation and mass loading of
fine aerosol particles with aerodynamic
diameters between <50 and 1000 nm as a
function of particle size155. The original
ARI AMS utilizes a quadrupole mass
spectrometer (Q) with EI ionization and
produces ensemble average data of particle
properties. Later versions employ time-offlight (ToF) mass spectrometers and can
produce complete mass spectral data for
single particles (153).
Despite the increasing amount of data
provided by both laboratory and field
studies, the nature and role of aerosol
particles responsible for health effects, as
well as of gaseous mixtures especially in
urban areas, is still a matter of debate. In
the former case the issue is of importance
also because the different constituents of
CNR Environment and Health Inter-departmental Project
the aerosol exhibit distinct sources and
emission/formation processes. Therefore,
linking toxicological and epidemiological
impacts of atmospheric particulate matter
to chemical composition is a key for the
evaluation of effective pollution abatement
Results of size-segregated aerosol
chemical analyses for Italian stations have
already been published during the last
ten years and are available in the peerreviewed literature and in project reports
(40). These data generally refer to sparse
measurements employing multi-stage
impactors in both urban (e.g, Bologna,
Catania, Rome) and rural/background
sites (e.g, Monte Cimone); information
on the inorganic and organic composition
of ultrafine to coarse particles have been
retrieved by chemical determinations of
size-segregated fractions.
Nevertheless, an increasing series of data
on the aerosol chemical composition
and size-distribution has been provided
by short-term intensive field studies
performed in the frame of national and
European research projects(46). During
instrumentation has been deployed for
aerosol characterization.
In this direction, the “Pilot study for
the assessment of health effects of the
chemical composition of ultrafine and
fine particles in Italy” project, recently
approved by CNR, will combine the results
of two advanced activities in the field of
atmospheric ultrafine particles composition
and their toxicological properties, carried
out by CNR-ISAC and CNR-IIA, with
two new advanced health studies carried
out by CNR-IFC and CNR-IBIM, aimed
at exploring short-term effects due to air
pollutants exposure in subjects with preexistent arrhythmia and lung diseases.
As far as CNR-ISAC and CNR-IIA are
concerned, in the project a collection of all
these data will be performed and a data-base
of size-resolved chemical compositions
of the aerosol classified according to site
characterization and sampling period
will be provided as outcome. The databases emerging from the above activities
will be further integrated and the results
evaluated to derive conclusions on the
available knowledge on the size-segregated
chemical composition of the aerosol in the
different environments explored. In this
view, a further step forward will be also
to identify systematic behaviours in the
contributions of inorganic and organic
chemical constituents as a function of
particles size, and depending on site
classification (urban, sub-urban, rural,
marine, high-altitude, etc.), as well as to
provide a summary of the constituents
of ultrafine particles. This will help
interpreting clinical and epidemiological
observations under an enhanced awareness
of the behaviour of particulate pollutants
in different environments. Finally, a
comparison with published results of
analogous measurements performed in
other European countries will be carried
out, to identify singularities to be further
investigated in the future. New directions
of research in the field of understanding
impact routes of ambient and indoor air on
health relate to selected species showing
a precise toxicant action and to their
size-segregated behaviour in aerosols. In
this view, research activities will be run
concerning either the characterization
or the toxicant and reactive behaviour
of the water-soluble organic fraction of
PM. Besides transition metals and PAHs,
on which some peer-reviewed literature
already exist, the chemical analysis of fine
particulate samples has shown that even
in urban areas the water-soluble fraction
of the aerosol contains large amounts of
Role of Atmospheric Pollution on Harmful Health Effects
poorly-characterized organic compounds
(WSOC, “water-soluble organic carbon”),
in contrast to the paradigm of many
toxicological studies which attributes
the organic-soluble and water-soluble
fractions of the aerosol to organic and
On the contrary, recent findings point
to WSOC as a major agent for aerosol
toxicity and oxidizing properties (56,116).
Although a number of bioassays have been
adopted in previous studies to provide
fast and sensitive measurement of the
aerosol chemical reactivity, mechanistic
pathways for toxicity were not established.
Relevant bioassays will be thus tested,
for the scopes of evaluating the oxidative
potential of airborne aerosol in humans
and animals. Among possible bioassays,
those will be selected which are sensitive
to reactive oxygen species, like superoxide
and hydrogen peroxide. The latter species,
indeed, can be produced in biological
liquids and tissues by organic compounds
via redox reactions. Such tests include
for instance those based on dithiothreitol
(DTT) consumption rate, or employing
dichlorofluorescin (DCFH) (56,156). The
key importance of testing these methods
to provide, i.e, an optimal application to
the analysis of the water-soluble organic
extracts of ambient aerosol samples is
a matter of evidence. Finally, a stateof-the-art analytical technique like the
Aerosol Mass Spectrometry (AMS) will
be employed for the quantitative mass
determination of UFPs. This is at date
an obligate step towards enhancing the
knowledge about responsibilities of the
atmospheric pollution on health impacts.
Indeed, the AMS is currently the only
technique providing unique information
on the short-term processes controlling the
concentration and composition of ultrafine
particles and their interaction with larger
particles. Moreover, the data analysis of
the emerging results from AMS will allow
comparison with the more consolidated
outcomes from available measurements by
multi-stage impaction methods.
In summary, by examining the priorities
for the evaluation of upcoming research
activities of CNR for linking atmospheric
aerosols composition and properties to
their health effects, at least two specific
key issues can already be addressed and
dedicated to a) ultrafine particles and b)
Keywords: PM, UFP, WSOC, AMS, oxidizing
Decision No. 1600/2002/EC of the
European Parliament and of the Council
of 22 July 2002 laying down the Sixth
Programme. Official Journal of the
European Communities, 2002;242:1–15.
The Clean Air for Europe (CAFE)
programme: towards a thematic strategy
for air quality. Brussels, European
Commission, 2001 (COM(2001)245).
WHO Regional Office for Europe. Health
aspects of air pollution with particulate
matter, ozone and nitrogen dioxide: report
on a WHO working group, 2003 Jan 13-15;
Bonn, Germany. WHO Regional Office
for Europe Available from: URL: http://
Pope CA. Lung cancer, cardiopulmonary
mortality, and long-term exposure to fine
particulate air pollution. J Am Med Assoc
Burden of Disease project. 2004 May 20.
w w w3.w h o. i n t / w h o s i s / m e n u
Amann M, Cofala J, Heyes C, Klimont
Z, Schöpp W. The RAINS model: a tool
for assessing regional emission control
strategies in Europe. Pollut Atmos 1999
CNR Environment and Health Inter-departmental Project
Amann M, Johansson M, Lükewille A,
Schöpp W, ApSimon H, Warren R, et al.
An integrated assessment model for fine
particulate matter in Europe. Water Air
Soil Pollut 2001;130:223–228.
Amann M, Cofala J, Heyes C, Klimont
Z, Mechler R, Posch M, et al. RAINS
REVIEW 2004. The RAINS Model.
Documentation of the Model Approach
Prepared for the RAINS Peer Review
2004; IIASA, Laxenburg, Austria.
Amman M, Berttok I, Cofala J, Gyarfas
F, Heyes C, Klimont Z, et al. Baseline
Scenarios for the Clean Air for Europe
Istitute for Applied Systems Analysis:
2005; Laxemburg, Austria.
Vialetto G, Contaldi M, De Lauretis
R, Lelli M, Mazzotta V, Pignatelli T.
Emission scenarios of air pollutants in
Italy using integrated assessment models.
Pollut Atmos 2005;185,71–78.
D’Elia I, Bencardino M, Ciancarella L,
Contaldi M, Vialetto G. Technical and
Non Technical Measures for Air Pollution
Emission Reduction: the Integrated
Assessment of the Regional Air Quality
Management Plans through the Italian
National Model. Atmos Environ
Pignatelli T, Bencardino M, Ciancarella
L, D’Elia I, Racalbuto S, Vialetto G
et al. Comparative and qualitative
analysis of impact scenarios developed
by RAINS_Europe and RAINS_Italy,
in the perspective of downscaling. In:
Anderssen RS, Braddock RD, Newham
LTH, editors. MODSIM09. Proceedings
of the 18th World IMACS Congress and
International Congress on Modelling
and Simulation. 2009 Jul 13-17; Cairns,
Australia. 2321-7.
Burke JM, Zufall MJ, Özkaynak H. A
population exposure model for particulate
matter: case study results for PM2.5
in Philadelphia, PA. J Exposure Anal
Environ Epidemiol 2001;11:470-489.
Position Paper on Ambient Air
Pollution: Carbon Monoxide. European
Commission, DG Environment, 1999.
16. Raub JA, Mathieu-Nolf M, Hampson NB,
Thom SR. Carbon monoxide poisoning —
a public health perspective. Toxicology ,
2000, 145, 12000, 1 – 14.
17. Peters A, Dockery DW, et al. Particulate
Air Pollution and Nonfatal Cardiac
Events. Health Effects Institute (HEI)
Report, 2005, Vol. 124.
18. Sarnat HB. CNS malformations: Gene
locations of known human mutations.
European Journal of Paediatric Neurology,
2005, 9, 6, 427-431.
19. Position Paper on Ambient Air
Pollution: Carbon Monoxide. European
Commission, DG Environment, 1997.
20. EPA (United States Environmental
Protection Agency). Integrated Science
Assessment for Oxides of Nitrogen —
Health Criteria. Annexes. Report. 2008,
21. Position Paper on Ozone. Ad-Hoc Working
Group on Ozone Directive and Reduction
Strategy Development of the European
Commission, DG Environment, 1999.
22. HEI Air Toxics Review Panel. MobileSource Air Toxics: A Critical Review of
the Literature on Exposure and Health
Effects. Health Effects Institute (HEI)
Report, 2008, Vol. 16.
23. Zielinska B. Atmospheric transformation
of diesel emissions. Experimental and
Toxicologic Pathology, 2005, 57, 1, 3142.
24. Carslaw N. A new detailed chemical
model for indoor air pollution. Atmos.
Environ. 2007, 41, 6, 1164-1179.
25. Martuzzi M, Mitis F, Iavarone I et al.
Health impact of PM10 and Ozone in
13 italian cities. WHO (World Health
Organization Regional Office for Europe),
ISBN 92 890 2293 0 WHOLIS number
26. Council Directive 96/62/EC of 27
September 1996 on ambient air quality
assessment and management. Official
Journal L 296 , 21/11/1996 P. 0055 –
27. UNECE Convention on Long-range
Transboundary Air Pollution (LRTAP).
Role of Atmospheric Pollution on Harmful Health Effects
World Health Organization (WHO).
Health aspects of air pollution. Report
E83080, 2004.
Forastiere F, Stafoggia M, Picciotto S,
Bellander T, D’Ippoliti D, Lanki T, von
Klot S, Nyberg F, Paatero P, Peters A,
Pekkanen J, Sunyer J, Perucci CA. A
case-crossover analysis of out-of-hospital
coronary deaths and air pollution in
Rome, Italy. Am J Respir Crit Care Med
2005; 172; 1549-1555.
Von Klot S, Peters A, Aalto P et al.
Health Effects of Particles on Susceptible
Subpopulations (HEAPSS) Study Group.
Ambient air pollution is associated
with increased risk of hospital cardiac
readmissions of myocardial infarction
survivors in five European cities.
Circulation, 2005; 112; 3073-3079.
Laden F, Neas, L.M, Dockery, D.W,
Schwartz J. Association of fine particulate
matter from different sources with daily
mortality in six US cities. Envir. Health.
Persp, 2000; 108; 941-947.
McCreanor J, Cullinan P, Nieuwenhuijsen
M.J et al. Respiratory effects of exposure
to diesel traffic in persons with asthma.
N. Engl. J. Med, 2007; 357; 2348-2358.
Lippmann M, Frampton M, Schwartz J
et al. The U.S. Environmental Protection
Agency Particulate Matter Health Effects
Research Centers Program: A Midcourse
Report of Status, Progress, and Plans.
Envir. Health. Persp. 2003; 111; 10741092
Russell A.G and Brunekreef B. A focus
on particulate matter and health. Environ.
Sci. Technol. 2009; 43; 4620 – 4625.
Samet J.M, Dominici F, Curriero F.C,
Coursac I, Zeger S.L. Fine particulate air
pollution and mortality in 20 US cities,
1987-1994. New Eng. J. Medicine, 2000;
343; 1742-1799.
Astolfi M.L, Canepari S, Catrambone M,
Perrino C and Pietrodangelo A. Improved
characterisation of inorganic components
in airborne particulate matter. Environ.
Chem. Letters, 2006; 3; 186-191.
37. Baltensperger U, Dommen J, Alfarra
M.R et al. Combined determination of
the chemical composition and of health
effects of secondary organic aerosols: The
POLYSOA project. Journal of Aerosol
Medicine and Pulmonary Drug Delivery,
2008; 21; 145 – 154.
38. Canepari, S, Perrino, C, Olivieri, F,
Astolfi, M. L. Characterisation of the
traffic sources of PM through sizesegregated sampling, sequential leaching
and ICP analysis. Atmos Environ, 2008;
42; 8161-8175.
39. Chan, Y.C, Simpson, R.W, McTainsh,
G.H. and Vowles, P.D. Characterisation
of chemical species in PM2.5 and
PM10 aerosols in Brisbane, Australia.
Atmospheric Environment 1997; 31;
40. Fabiani R, De Bartolomeo A, Rosignoli
P, Morozzi G, Cecinato A, Balducci
characterization of airborne total
suspended particulate and PM10
organic extracts. Polycyclic Aromatic
Compounds, 2008; 28; 486-499.
41. Matta, E, Facchini M.C, Decesari S,
Mircea M, Cavalli F, Fuzzi S, Putaud
J-.P,. Dell’Acqua A. Mass closure on
the chemical species in size-segregated
atmospheric aerosol collected in a urban
area of the Po Valley, Italy. Atmos. Chem.
Phys. 2003; 3; 623 - 637.
42. Perrino C, Canepari S, Cardarelli E,
Catrambone M, Sargolini, T. Inorganic
costituents of urban air pollution in the
Lazio region (Central Italy). Environ.
Monit. Assess. 2007; 128; 133-151.
43. Perrino C, Canepari S, Catrambone
M, Dalla Torre S, Rantica E, Sargolini
T. Influence of natural events on
the concentration and composition
of atmospheric particulate matter.
Atmospheric Environment 2009; 43;
44. Putaud, J.P, Raes F, Van Dingenen R. et al.
A European aerosol phenomenology-2 :
chemical characteristics of particulate
matter at kerbside, urban, rural and
background sites in Europe. Atmospheric
CNR Environment and Health Inter-departmental Project
Environment, 2004; 38; 2579-2595.
45. Querol, X, Alasturey A, Ruiz C.R, et al.
Speciation and origin of PM10 and PM2.5
in selected European cities. Atmospheric
Environment. 2004;38; 6547-6555.
46. Vecchi R, Marcazzan G, Valli G. A
study on nighttime–daytime PM10
concentration and elemental composition
in relation to atmospheric dispersion in the
urban area of Milan (Italy). Atmospheric
Environment. 2007; 41; 2136–44.
47. Canepari, S, Pietrodangelo A, Perrino C,
Astolfi M.L, Marzo M.L. Enhancement
of source traceability of atmospheric
PM by elemental chemical fractionation.
Atmospheric Environment, 2009; 43;
48. Snyder, D. C, Schauer, J. J, Gross, D. S,
Turner, J. R. Estimating the Contribution
of Point Sources to Atmospheric Metals
using Single-Particle Mass Spectrometry.
Atmospheric Environment, 2009; 43;
49. Viana M,. Kuhlbusch T.A.J, Querol X, et
al. Source apportionment of particulate
matter in Europe: A review of methods
and results. Aerosol Science 2008; 39;
50. Branis M, Safranek J, Hytychova
A. Exposure of children to airborne
particulate matter of different size
fractions during indoor physical education
at school. Building and Environment,
2009; 44; 1246–1252.
51. Chan A.T, Chung M.W. Indoor–outdoor
air quality relationships in vehicle: effect
of driving environment and ventilation
modes. Atmospheric Environment, 2003;
37; 3795-3808.
52. Chao C.Y.H, Tung T.C. An empirical model
for outdoor contaminant transmission into
residential buildings and experimental
verification. Atmospheric Environment,
2001; 35; 1585-1596.
53. Poupard O, Blondeau P, Iordache V,
Allard F. Statistical analysis of parameters
influencing the relationship between
outdoor and indoor air quality in schools.
Atmospheric Environment, 2005; 39;
54. Rudel R.A, Perovich L.J. Endocrine
disrupting chemicals in indoor and
outdoor air. Atmospheric Environment,
2009; 43; 170-181.
55. Saliba N.A, Atallah M, Al-Kadamany G.
Levels and indoor–outdoor relationships
of PM10 and soluble inorganic ions in
Beirut, Lebanon. Atmospheric Research,
2009; 92; 131-137.
56. Tippayawong N, Khuntong P, CNitatwichit
C, Khunatorn Y, Tantakitt C. Indoor/
outdoor relationships of size-resolved
particle concentrations in naturally
ventilated school environments. Building
and Environment, 2009; 44; 188–197.
57. Biswas S, Verma V, Schauer J, Cassee
F, Cho A, and Sioutas C. Oxidative
potential of semi-volatile and non volatile
particulate matter (PM) from heavy-duty
vehicles retrofitted with emission control
technologies. Environ. Sci. Technol,
2009; 43; 3905 – 3912.
58. Binkovà B, Vesely D, Veselà D,
Jelinek R, Sram R.J. Genotoxicity and
embryotoxicity of urban air particulate
matter collected during winter and
summer period in two different districts
of the Czech Republic. Mutation Research
Genetic Toxicology and Environmental
Mutagenesis 1999; 440; 45-58.
59. Calderón-Garcidueñas L, Solt A.C,
Henríquez-Roldán C, Torres-Jardón R,
Nuse B, Herritt L, Villarreal-Calderón R,
Osnaya N, Stone I, García R, Brooks D.M,
González-Maciel A, Reynoso-Robles R,
Delgado-Chávez R, Reed W. Long-term
air pollution exposure is associated with
neuroinflammation, an altered innate
immune response, disruption of the
blood-brain barrier, ultrafine particulate
deposition, and accumulation of amyloid
beta-42 and alpha-synuclein in children
and young adults. Toxicol Pathol, 2008;
36; 289-310.
60. Geller M.D, Ntziachristos L, Mamakos
A, Samaras Z, Schmitz D. A. , Froines
J.R, Sioutas C,. Physicochemical and
redox characteristics of particulate matter
(PM) emitted from gasoline and diesel
passenger cars. Atmospheric Environment
Role of Atmospheric Pollution on Harmful Health Effects
2006; 40; 6988-7004.
61. Risom L, Møller P, Loft S. Oxidative
stress-induced DNA damage by particulate
air pollution. Mutation Research, 2005;
59; 119–137.
62. Rückerl R, Ibald-Mulli A, Koenig W,
Schneider A, Woelke G, Cyrys J, Heinrich
J, Marder V, Frampton M, Wichmann H.E,
Peters A. Air pollution and markers of
inflammation and coagulation in patients
with coronary heart disease. Am J Respir
Crit Care Med, 2006 15; 173; 432-441.
63. Duker A.A, Carranza E.J.M, Hale M.:
Arsenic geochemistry and health. Environ
Int 2005;31: 631641.
64. Sanchez-Rodas D, Sanchez de la
Campa A.M, de la Rosa J, et al. Arsenic
speciation of atmospheric particulate
matter (PM10) in an industrialised urban
site in southwestern Spain. Chemosphere
65. Thomaidis N.S,. Bakeas E.B, Siskos
P.A. Characterization of lead, cadmium,
arsenic and nickel in PM2.5 particles
in the Athens atmosphere, Greece.
Chemosphere 2003;52:959966.
66. Kanias G.D, Viras L.G, Grimanis A.P.
Source identification of trace elements
emitted into Athens atmosphere, Relation
between trace elements and tropospheric
ozone. J Radioanal Nucl Chem
67. Güllü G, Dogan G, Tuncel G.
Atmospheric trace elements and major
ion concentrations over the eastern
Mediterranean Sea: Identification of
anthropogenic source regions. Atmos
Environ 2005;39:63766387.
68. Pacyna J, Pacyna E.G. An assessment
of global and regional emissions of
trace metals to the atmosphere from
Environ Res 2001;9:269298.
69. Flament P, Bertho M.L, Deboudt K,
Véron A, Puskaric E. European isotopic
signatures for lead in atmospheric
aerosols: a source apportionment based
upon 206Pb/207Pb ratios. Sci Total Environ
70. Pirrone N, Costa P, Pacyna J.M. Past,
current and projected atmospheric
emissions of trace elements in the
Mediterranean region. Water Sci Technol
Pirrone N, Allegrini I, Keeler G.J, Nriagu
J.O, Rossmann R, Robbins J.A. Historical
and Depositions in North America
Compared to Mercury Accumulations
in Sedimentary Records. Atmos Environ
Pirrone N, Costa P, Pacyna J. Past,
Current and Projected Emissions of Trace
Elements in the Mediterranean Basin.
Water Sci Technol 1999;39:1-7.
Lindqvist O, Johansson K, Aastrup M,
Andersson A, Bringmark L, Hovsenius G,
et al. Mercury in the Swedish environment
- recent research on causes, consequences
and corrective methods. Water Air Soil
Pollut 1991;55.
Pirrone N, Costa P, Pacyna J.M, Ferrara
R. Atmospheric mercury emissions
from anthropogenic sources in the
Mediterranean Region. Atmos Environ
Sprovieri F, Hedgecock I.M, Pirrone N. An
Investigation of the Origins of Reactive
Gaseous Mercury in the Mediterranean
Marine Boundary Layer Atmos Chem
Phys Discuss 2009;9:24815-24846.
Pacyna E.G, Pacyna J.M, Pirrone N.
European emissions of atmospheric
mercury from anthropogenic sources in
1995. Atmos Environ 2001;35:2987-2996.
Bindler R, Renberg I, Appleby P.G,
Anderson N.J, Rose N.L. Mercury
accumulation rates and spatial patterns
in lake sediments from west Greenland: a
coast to ice margin transect. Environ Sci
Technol 2001;35:736–741.
Biester H, Kilian R, Franzen C, Woda
C, Mangini A, Schöler H.F. Elevated
mercury accumulation in a peat bog
of the Magellanic Moorlands, Chile
(538S)—An anthropogenic signal from
the Southern Hemisphere. Earth Planet
Sci Lett 2002;201:609–620.
Lamborg C.H, Fitzgerald W.F, Damman
A.W.H,. Benoit J.M, Balcom P.H,
CNR Environment and Health Inter-departmental Project
Engstrom D.R. Modern and historic
atmospheric mercury fluxes in both
hemispheres: global and regional mercury
cycling implications. Global Biogeochem
Cycles 2002;16:104.
Lindberg S, Bullock R, Ebinghaus R, et al.
A synthesis of progress and uncertainties
in attributing the sources of mercury in
deposition. Acta Biomate 2007;36(1):1932.
US-EPA (U.S. Environmental Protection
Agency). Mercury Report to Congress,
Vol VI: Characterization of Human Health
and Wildlife Risks from anthropogenic
Mercury Emissions in the United States.
EPA-452/R-97-001f. Washington. DC.
U.S. Environmental Protection Agency,
Schroeder W.H, Munthe J. Atmospheric
Mercury – An Overview. Atmos Environ
Sprovieri F, Pirrone N, Gärdfeldt K,
Sommar J. Mercury speciation in the
marine boundary layer along a 6000 km
cruise path around the Mediterranean
Sea. Atmos Environ 2003; 37:63-71.
Pirrone N, Ferrara R, Hedgecock I.M, et
al. Dynamic Processes of Mercury Over
the Mediterranean Region: results from
the Mediterranean Atmospheric Mercury
Cycle System (MAMCS) project. Atmos
Environ 2003;37(1):21-39.
Bornmann G, Henke G, Alfes H,
Mollmann H. Intestinal absorption of
metallic mercury (in German) Arch
Toxicol 1970;26:203-209.
Hursh J.B. Partition coefficients of
mercury (203-Hg vapor between air
and biological fluids. J Appl Toxicol
Berlin M, Friberg L, Nordberg G.F, Vouk
V.B. editors, Mercury In: Handbook on
the toxicology of Metals, pp. 387-445, 2nd
ed. New York: Elsevier; 1986.
WHO. Environmental health criteria
101: Methylmercury. World Health
Organisation Geneva, International
Programme on Chemical Safety, 1990.
WHO. Environmental health criteria
118: Inorganic mercury. World Health
Organisation Geneva, International
Programme on Chemical Safety 1991.
WHO. Joint FAO/WHO expert committee
on food additives. Fifty-third meeting,
1999 Jun 1-10, Rome, Italy. Summary and
conclusions. Available at.
U.S. EPA (1997) Mercury Report to
Congress. U.S. Environmental Protection
Research Triangle Park, NC.
Clarkson T.W. The toxicology of mercury.
Crit Rev Clin Lab Sci 1997;34:369-403.
Aberg B, Ekman L, Falk R, Greitz U,
Persson G, Snihs J.O. Metabolism of
methylmercury (203 Hg) compounds in
man. Arch Environ Health 1969;19:478484.
Finlayson-Pitts B.J, Pitts J.N. Jr. Chemistry
of the upper and lower atmosphere. San
Diego CA (USA), Academic Press, 2000.
Pankow J.F, Luo W, Bender D.A, Isabelle
L.M, Hollingsworth J.S, Chen C, Asher
W.E, Zogorski J.S. Concentrations and
co-occurrence correlations of 88 volatile
organic compounds (VOCs) in the ambient
air of 13 semi-rural to urban locations in
the United States. Atmos. Environ. 2003;
37; 5023-5046.
Ciccioli P, Cecinato A, Brancaleoni E,
Frattoni M. Identification and quantitative
evaluation of C4-C14 volatile organic
compounds in some urban, suburban and
forest sites in Italy. Fresenius Envir. Bull.
1992; 1; 73-78.
Ciccioli P, Brancaleoni E, Cecinato
A, Sparapani R. Identification and
compounds in forest areas of Northern
and Southern Europe and a remote site of
the Himalaya region by high-resolution
gas chromatography - mass spectrometry.
J. Chromatog. 1993; 643; 55-69.
Campbell N.J, McCulloch A. Climate
change implications of manufacturing
refrigerants: a Calculation of ‘production’
energy contents of some common
refrigerants. Process Safety and Environ.
Protect. 1998; 76; 239-244.
Role of Atmospheric Pollution on Harmful Health Effects
100. Highwood E.J, Shine K.P, Hurley M.D,
Wallington T.J. Estimation of direct
radiative forcing due to non-methane
hydrocarbons. Atmos. Environ. 1999; 33;
101. Rodriguez J.M. Probing stratospheric
ozone. Science 1993; 261; 1128-1129.
102. Rowland F.S. Atmospheric chemistry:
causes and effects. J. Mar. Technol. Soc.
1991; 25; 12-18.
103. Derwent R.G, Jenkin M.E, Saunders
S.M. Photochemical ozone creation
potentials for a large number of reactive
hydrocarbons under European conditions.
Atmos. Environ. 1996; 30; 181-199.
104. Croute F, Poinsot .J, Gaubin Y, Beau B,
Simon V, Murat J.C, Soleilhavoup J.P.
Volatile organic compounds cytotoxicity
and expression of HSP72, HSP90 and
GRP78 stress proteins in cultured human
cells. Biochim. Biophys. Acta (BBA)/
Molec. Cell Res. 2002; 1591; 147-155.
105. European Parliament and Council.
Directive 96/62/EC of 27 September 1996
on Ambient Air Quality Assessment and
Management. Strasbourg, 1996.
106. European Parliament and Council.
Directive 2000/69/EC of 16 November
2000 relating to Limit Values for Benzene
and Carbon Monoxide in Ambient air.
Strasbourg, 2000.
107. European Parliament and Council.
Directive 2001/80/EC of 23 October 2001
on Limitation of Emissions of Certain
Pollutants into the Air from Large
Combustion Plants. Strasbourg, 2001.
108. European Parliament and Council.
Directive 2001/81/EC of 23 October 2001
on National Emission Ceilings for Certain
Atmospheric Pollutants. Strasbourg,
109. European Parliament and Council.
Directive 2002/3/EC of 12 February
2002 Relating to Ozone in Ambient Air.
Strasbourg, 2002.
110. European Parliament and Council.
Directive 2002/45/EC of 25 June 2002
amending the Council Directive 76/769/
EEC relating to Restrictions on the
Marketing and Use of Certain Dangerous
Substances and Preparations (short-chain
chlorinated paraffins). Strasbourg, 2002.
European Parliament and Council
(2003). Directive 2003/87/EC of the of
13 October 2003 establishing a Scheme
for Greenhouse Gas Emission Allowance
Trading within the Community and
amending Council Directive 96/61/EC.
Strasbourg, 2003.
European Parliament and Council.
Commission Decision 2004/297/EC of
19 March 2004 concerning guidance
for implementation of Directive 2002/3/
EC of the European Parliament and the
Council relating to Ozone in Ambient
Air. Strasbourg, 2004.
European Parliament and Council.
Directive 2004/101/EC of 27 October
2004 amending Directive 2003/87/EC
establishing a Scheme for Greenhouse
Gas Emission Allowance Trading
within the Community, in Respect of the
Kyoto Protocol’s Project Mechanisms.
Strasbourg, 2004.
Italian Ministry of the Environment.
Ministry Decree of 25 November 1994
relative to Updating of the technical rules
concerning the concentration limits and
the precautionary and warning levels
regarding the atmospheric pollutants
in the urban areas and indications for
the measure of some pollutants referred
in the Ministerial released on April
15th. Gazzetta Ufficiale Italiana della
Repubblica Italiana, Suppl. No. 290, 13
December 1994. Rome, 1994.
Alves C, Pio C, Duarte A. Composition of
extractable organic matter of air particles
from rural and urban Portuguese areas.
Atmos. Environ. 2001; 35; 5485-5496.
Simoneit B.T.R. Organic matter in the
troposphere – III – Characterization
and sources of petroleum and pyrogenic
residues in aerosols over the western
United States. Atmos. Environ.1984; 18;
Simoneit B.R.T, Mazurek M.A. Organic
matter of the troposphere-II. Natural
background of biogenic lipid matter in
aerosols over the rural Western United
CNR Environment and Health Inter-departmental Project
States. Atmos. Environ. 1982; 16; 21392159.
118. Baltensperger U, Dommen J, Alfarra
M.R, et al. Combined determination of
the chemical composition and of health
effects of secondary organic aerosols:
The POLYSOA project. J. Aerosol Med.
Pulmonary Drug Deliv. 2008; 21; 145 –
119. Binkovà B, Vesely D, Veselà D, Jelinek R,
Sram R.J. Genotoxicity and embryotoxicity
of urban air particulate matter collected
during winter and summer period in two
different districts of the Czech Republic.
Mutat. Res. Genetic Toxicol. Environ.
Mutagen. 1999; 440; 45-58.
120. Fabiani R, De Bartolomeo A, Rosignoli
P, Morozzi G, Cecinato A, Balducci
characterization of airborne total
suspended particulate and PM10 organic
extracts. Polycyc. Aro. Comp. 2008; 28;
121. Hannigan M.P, Cass G.R, Penman B.W,
et al. Bioassay-directed chemical analysis
of Los Angeles airborne particulate
matter using a human cell mutagenicity
assay. Environ. Sci.Technol. 1998; 32;
122. Cass G.R. Organic molecular tracers for
particulate air pollution sources. Trends
in Analyt. Chem. 1998; 17; 356-366.
123. Kavouras I.G, Khalili N.R, Scheff P.A,
Holsen T.M. PAH source fingerprints for
coke ovens, diesel and gasoline engines,
highway tunnels and wood combustion
emissions. Atmos. Environ. 1995; 29;
124. Kamens R.M, Guo Z, Fulcher J.N,
Bell D.A. The influence of humidity,
sunlight, and temperature on the daytime
decay of polyaromatic hydrocarbons on
atmospheric soot particles. Environ. Sci.
Technol. 1988; 22; 103–108.
125. International Agency for Research on
Cancer IARC. Polynuclear aromatic
environmental and experimental data.
Monographs on the evaluation of
carcinogenic risk of chemicals to humans,
vol. 32. Lyon (F), IARC, 1983.
126. Claxton L.D, Matthews P.P, Warren S.H.
The genotoxicity of ambient outdoor air, a
review: Salmonella mutagenicity. Mutat.
Res. 2004; 567, 347–399.
127. Turpin B. J, Saxena P, Andrews E.
Measuring and simulating particulate
organics in the atmosphere: problems
and prospects. Atmos. Environ. 2000; 34;
128. Chrysikou L.P, Samara C.A. Seasonal
variation of the size distribution of urban
particulate matter and associated organic
pollutants in the ambient air. Atmos.
Environ. 2009; 43; 4557-4569.
129. Tremblay R.T, Riemer D, Zika R.G.
Organic composition of PM2.5 and
size-segregated aerosols and their
sources during the 2002 Bay Regional
Atmospheric Chemistry Experiment
(BRACE), Florida, USA. Atmos. Environ.
2007; 41; 4323-4335.
130. European Parliament and Council.
Directive 2004/107/EC of 15 December
2004 relating to Arsenic, Cadmium,
Mercury, Nickel and Polycyclic Aromatic
Hydrocarbons in Ambient Air. Strasbourg,
131. European Parliament and Council.
Directive 2008/1/EC of 15 January
2008 concerning Integrated Pollution
Prevention and Control. Strasbourg,
132. European Parliament and Council.
Directive 2008/50/EC of 21 May 2008 on
Ambient Air Quality and Cleaner Air for
Europe. Strasbourg, 2008.
133. Italian Ministry of the Environment. Law
by Decree No. 152 of 3 August 2007 on
“Implementation of the 2004/107/CE
Directive concerning arsenic, cadmium,
mercury, nickel and polycyclic aromatic
hydrocarbons in ambient air”. Gazzetta
Ufficiale della Repubblica Italiana No.
213, 13 September 2007, Suppl. No. 194.
Rome, 2007.
134. NNI (National Nanotechnology Initiative).
2004. What Is Nanotechnology?
Role of Atmospheric Pollution on Harmful Health Effects
135. Kulmala H, Vehkamaki H, Petaja T,
Dal Maso M, Lauri A, Kerminen V.-M,
Birmili W, McMurry P.H. Formation and
growth rates of ultrafine atmospheric
particles: a review of observations. J.
Aerosol Sci, 2004, 35, 143-176.
136. Fushimi A, Hasegawa S, Takahashi
K, Fujitani Y, Tanabe K, Kobayashi S.
Atmospheric fate of nuclei-mode particles
estimated from the number concentrations
and chemical composition of particles
measured at roadside and background
sites. Atmos Environ. 2008;42:949–59.
137. Hirano S, Nitta H, Moriguchi Y,
Kobayashi S, Kondo Y, Tanabe K,
et al. Nanoparticles in emissions and
atmospheric environment: now and
future. J Nanopart Res. 2003;5:311–21.
138. Hirano S. A current overview of health
effect research on nanoparticles. Environ
Health Prev Med (2009) 14:223–225.
139. Zhou J, Swietlicki E, Hansson H.C, Artaxo
P. Aerosol particle size distribution and
hygroscopic growth in the Amazonian
rain forest. J. Aerosol Sci, 1999, 30, S163S164.
140. Bower K.N, Beswick K.M, Burgess R.;
Stromberg I.M, Gallagher M.W. Aerosol,
trace gas and thermal gradient structure of
urban conurbations measured by aircraft.
J. Aerosol Sci, 2000, 31, 114-115.
141. Ramanathan, R. A note on the use
of the analytic hierarchy process for
environmental impact assessment. J.
Environ. Management, 2001, 63, 27-35.
142. U.S. EPA. 2004. Air Quality Criteria for
Particulate Matter. Vol 3. 600/P-95-001cF.
Washington DC:U.S. Environmental
Protection Agency, Office of Research
and Development
143. Oberdörster G, Oberdörster E. and
Oberdörster J, 2005. Nanotoxicology: an
emerging discipline evolving from studies
of ultrafine particles. Review. Environ.
Health Perspect, 113 (7), 823 – 839.
144. McCreanor J, Cullinan P, Nieuwenhuijsen
M.J, Stewart-Evans J, Malliarou E, Jarup
L, Harrington R, Svartengren M. Han
I.K, Ohman-Strickland P, Chung K.F,
Zhang J. Respiratory effects of exposure
to diesel traffic in persons with asthma. N
Engl J Med; 2007;357:2348-2358.
145. Andersen Z.J, Loft S, Ketzel M, Stage M,
Scheike T, Hermansen M.N, Bisgaard H.
Ambient air pollution triggers wheezing
symptoms in infants. Thorax; 2008;
146. Rückerl R, Ibald-Mulli A, Koenig W,
Schneider A, Woelke G, Cyrys J, Heinrich
J, Marder V, Frampton M, Wichmann H.E,
Peters A. Air pollution and markers of
inflammation and coagulation in patients
with coronary heart disease. Am J Respir
Crit Care Med 2006; 15;173:432-441.
147. Calderón-Garcidueñas L, Solt A.C,
Henríquez-Roldán C, et al. Long-term
air pollution exposure is associated with
neuroinflammation, an altered innate
immune response, disruption of the
blood-brain barrier, ultrafine particulate
deposition, and accumulation of amyloid
beta-42 and alpha-synuclein in children
and young adults. Toxicol Pathol,
148. Forastiere F, Stafoggia M, Picciotto S, et
al. A case-crossover analysis of out-ofhospital coronary deaths and air pollution
in Rome, Italy. Am J Respir Crit Care
149. Viana M,. Kuhlbusch T.A.J, Querol X,
Alastuey A, Harrison R.M, Hopke P.K,
et al. Source apportionment of particulate
matter in Europe: A review of methods
and results. Aerosol Science, 2008;39,
827 – 849.
150. Chow J.C, Watson J.G, Kuhns H,
Etyemezian V. et al. Source profiles for
industrial, mobile, and area sources in
the Big Bend Regional Aerosol Visibility
and Observational study. Chemosphere,
2004;54 (2), 185 – 208.
151. Morozzi G, Mastrandrea V, Trotta F. et al.
Chemical characterization and biological
properties of airborne particulate matter.
Aerobiologia, 1992, 8, 451-457.
152. Fabiani R, De Bartolomeo A, Rosignoli
P, Morozzi G. et al. Chemical and
toxicological characterization of airborne
total suspended particulate and PM10
organic extracts. Polycyclic Aromatic
CNR Environment and Health Inter-departmental Project
Compounds, 2008, 28, 486-499
153. Peters A, Dockery D.W, et al. Particulate
Air Pollution and Nonfatal Cardiac
Events. Health Effects Institute (HEI)
Report, 2005, Vol. 124.
154. Maynard A.D. The development of a new
thermophoretic precipitator for scanning
transmission electron microscope analysis
of ultrafine aerosol particles. Aerosol Sci.
Technol, 1995, 23, 521-533.
155. Canagaratna M.R, Jayne J.T, Jimenez
J.L, et al. Chemical and microphysical
characterization of ambient aerosols with
the aerodyne aerosol mass spectrometer.
Mass Spectrometry Reviews, 2007, 26,
185– 222.
156. Ferge T, Muhlberger F, Zimmermann
R. 2005. Application of infrared laser
desorption vacuum-UV single-photon
ionization mass spectrometry for analysis
of organic compounds from particulate
matter filter samples. Anal Chem
157. Jayne J.T, Leard D.C, Zhang X, Davidovits
P, Smith K.A, Kolb C.E. Worsnop DR.
2000. Development of an aerosol mass
spectrometer for size and composition
analysis of submicron particles. Aerosol
Sci Technol 33:49–70
158. Venkatachari P, Hopke P.K, Grover B.
D. et al, 2005. Measurement of particlebound reactive oxygen species in
Rubidoux aerosols. J. Atmos. Chem. 50,
49 – 58.
Human Biomonitoring
E. Leonia,b, A. I. Scovassia
a. CNR, Institute of Molecular Genetics (IGM), Pavia, Italy
b. Salvatore Maugeri Foundation, Pavia
[email protected]
Human biomonitoring is the discipline devoted to the identification of biomarkers useful to measure
environmental exposure, to monitor its biological effects and causal relationship with pathological conditions,
and to possibly define the genetic susceptibility of the general population. The search for reliable biomarkers,
i.e. characteristics that are objectively measured and validated as health or disease requires the expertise of
scientists with diversified specializations, who are able to tackle problems of ever increasing complexity
using complementary approaches. Within the frame of the PIAS-CNR project “Environment and Health”, we
have constituted a Biomonitoring network, composed by highly qualified scientists from CNR, and external
teams. This action aims at promoting a scientific strategy to develop and validate biomarkers of effect,
exposure and susceptibility.
1.1 Background
Chemicals are present in the air, ingested
food and water, at the workplace as well
as at home; the exposure to them can
occur through inhalation, cutaneous
contact, and ingestion. The evaluation and
measurement of the impact of chemicals
on human health is achieved by “Human
Biomonitoring” (HBM), defined as the
“systematic standardized measurement of
substance or its metabolites in body fluids
(blood and urine) of exposed persons”
(1-4). The different steps of HBM ranging
from the exposure step to the health
impairment (and eventually disease
occurrence) are schematized in Figure 1.
The estimation of the dose really taken up
after the exposure (internal dose) can be
addressed by qualitative and quantitative
assays able to detect chemicals and/or
metabolites in biological fluids; this action
provides basic information to identify
exposure biomarkers.
The HBM survey is further extended
to the analysis of biochemical and
biological effects: as illustrated in
Figure 1, biochemical effects could be
monitored through the reaction of reactive
substances (or their metabolites) with
biological macromolecules, such as DNA
and proteins. The study of biological
effects induced by chemicals implies
cellular, cytogenetic, genetic, biochemical,
metabolic and immunological approaches.
This strategy aims at defining a panel
of effect biomarkers. Furthermore, it is
widely recognized that biomarkers of
susceptibility may influence each element
in the exposure-to-disease paradigm
(Figure 1). This class of biomarkers
refers to individual factors (e.g. genetic or
metabolic) that influence the sensitivity
to hazardous compounds. Markers that
are measurable at low exposure or dose,
have the greatest potential utility to
prevent disease, while later markers are
CNR Environment and Health Inter-departmental Project
Figure 1: The exposure-to-disease paradigm.
most closely related to the endpoint of risk about the link between blood mercury and
dioxin and fish diet (6). However, to better
exposure, that is disease development (5).
As discussed in the special issue of the exploit these results, a harmonised strategy
Bulletin Epidémiologique Hebdomadaire allowing comparisons between countries
(6), HBM can help set priorities for public is required.
health and regulatory follow-up. In fact, The selection of chemicals allowed to
periodic measurement of biomarkers in enter a biomonitoring program is very
the population reveals how body burdens complex. The Centre for Disease Control
of chemicals vary from season to season, and Prevention (CDC) has identified a
year to year and decade to decade; by number of important variables able to
comparing the results over the time it is influence the so-called process, including
possible to evaluate the trend of people’s the evidence of exposure, the presence
exposure to environmental chemicals. and significance of health effects after a
Large biomarker studies can highlight given level of exposure, the development
exposure differences among racial, of assays for accurately measuring
geographic or socioeconomic groups. As biological concentrations of the chemical
specified in the above document (6), it is agent, available specimens, in particular,
urgent to define the association between the blood and/or urine samples, and costlevel distribution of chemicals in humans and effectiveness.
geographic/social/demographic parameters, As reported in the literature, many
in order to generate exposure maps and to chemicals released into the environment can
promote better-targeted risk assessment disturb the development of the endocrine
and risk management actions. A number of system and of the organs that respond to
examples demonstrated that a policy change endocrine signals and can interfere with
can reduce people’s exposure to pollutants, the correct functioning of endocrine
e.g. the decision of way out leaded petrol organs. Such chemicals are generally
because of its established association with named “Endocrine Disruptors” (EDs) (7).
blood lead, the information campaigns EDs causing abnormalities and impaired
Human Biomonitoring
reproductive performance in some species,
are associated with changes in immunity
behaviour and skeletal deformities and are
responsible for apparent changes observed
in human health patterns over recent
decades, including an increased incidence
of certain types of hormone-sensitive
cancers (7,8).
1.2 State of the art
The exposure of the general population
to xenobiotics through different routes
is a matter of growing concern. To face
this problem, the European Commission
adopted in 2003 a Strategy on Environment
and Health (9). Then, the EU launched
the “2004-2010 Environment and Health
Action Plan”, designed to give the EU
the scientifically grounded information
to reduce the adverse health impacts of
certain environmental factors and to
endorse better cooperation among actors in
the environment, health and research fields
(10,11). The final goal of the Action Plan
is to promote and integrate environment
and health information, and to identify
emerging issues, reviewing and adjusting
risk reduction policies and improving
communication. Of course, this action
implies the development of a HMB policy
aiming at monitoring activities in human
beings, using biomarkers, which focus on
environmental exposures, diseases and/or
disorders and genetic susceptibility, and
their potential relationships (12).
The mid-term review of the action plan
pointed out the difficulty of collecting
and comparing HBM data from different
countries, given that implemented
methodologies and sample collection
protocols differ (13). In 2007, the EU Council
invited the Commission to ensure adequate
funding for the EU pilot project on HBM,
to pay attention to the existing regulatory
frameworks and, most important thing, to
demonstrate the added value of HBM as
policy tool and support to public health
interventions (14). Despite the big effort
of the EU action, experiences of HBM
programmes at country level reveal the
need for harmonisation. In this respect, last
December, in Brussels, 35 partners coming
from 27 European countries and including
governments, research institutes, the
Health and Environment Alliance (HEAL)
and the European Chemical Industry
Council (CEFIC) established a consortium
to perform human biomonitoring at
European scale (COPHES) (15). This
consortium aims at developing a functional
framework that contributes to the
definition, organization, and management
of a coherent approach towards HBM
in Europe, including strategies for data
interpretation and integration with
environmental and health data. A further
priority of COPHES is to regulate data
and to provide key information to all
stakeholders from the public to the policy
markers. The WHO 5th Environment and
Health Ministerial Conference (Parma,
March 10-12, 2010) is the next milestone
in the European environment and health
process, now in its twentieth year. Focused
on protecting children’s health in a
changing environment, the Conference
will drive Europe’s agenda on emerging
environmental health challenges for the
years to come (16).
HBM aims at estimating people’s exposure
to pollutants, thus providing information
about possible health effects and options
of policy measures to reduce exposure.
Persistent organic pollutants (POPs) are
chemical substances that persist in the
environment, bioaccumulate through the
food, and pose a risk of causing adverse
effects to human health and the environment.
Priority POPs include pesticides (such
CNR Environment and Health Inter-departmental Project
as DDT), industrial chemicals (such as
polychlorinated biphenyls, PCBs) and
unintentional by-products of industrial
processes such as dioxins (PCDDs) and
furans (PCDFs). Pesticides, PCBs, PCDDs,
PCDFs, and other emerging substances
have been detected in foodstuffs and
are potentially toxic to human health. In
addition to the diet, indoor and outdoor air
pollutants are possible sources of health
risk; however, available data are scarce and
inconclusive. POPs are transported across
international boundaries far from their
sources, even to regions where they have
never been used or produced. For most of
these chemicals, we simply do not know
how they pass through the environment,
whether they are accumulated, dispersed
or transformed, and how they affect living
organisms at different concentrations (17).
Consequently, POPs pose a threat to the
environment and to human health (18).
Among POPs, some trace elements (TEs)
e.g. cadmium, arsenic, lead, mercury
and nickel, are very dangerous for health
because they tend to bioaccumulate,
thus enhancing their concentration in a
biological organism over the time. TEs
may enter the human body through food,
water, air, or absorption through the skin
when they come in contact with humans
in agriculture and in manufacturing,
pharmaceutical, industrial, or residential
settings. Human exposure to TEs has
risen dramatically in the last 50 years as
a result of an exponential increase in their
use in industrial processes and products.
In general, TEs are systemic toxins
with specific neurotoxic, nephrotoxic,
and teratogenic effects; they can
induce impairment and dysfunction in
excretive organs (colon, liver, kidneys,
skin), endocrine and energy production
pathways, enzymatic, gastrointestinal,
immune, nervous, reproductive, and
urinary apparatus (19).
A special class of toxicants is represented by
Endocrine Disruptors (EDs). In 1991, at the
Wingspread Conference, expert scientists
at a work session on endocrine-disruptors
concluded that “Many compounds
introduced into the environment by human
activity are capable of disrupting the
endocrine system of animals, including
fish, wildlife, and humans. Endocrine
disruption can be profound because of the
crucial role hormones play in controlling
development” (20). EDs interfere with
the normal function of endocrine system
and can exert adverse effects on the
reproductive and other, indirectly related,
physiological systems.
EDs are natural or synthetic compounds,
derived primarily from anthropogenic
activities, ubiquitous in the environment.
Many of them are resistant to
biodegradation, due to their structural
stability, and persist in the environment.
EDs present in the environment include
a variety of potent human and veterinary
pharmaceutical products, personal care
products, nutraceuticals and phytosterols.
Substantial research has been carried
out on the mechanisms and effects of
EDs. Nevertheless it is still unclear to
what extent and/or in what situations and
population subgroups EDs may represent
a significant, long-term health risk.
To monitor the potential environmental
and health impacts of EDs, the EU adopted
a Communication to the Council and
European Parliament on a Community
Strategy for EDs in December 1999 (21). In
the proposal for a new policy for chemicals
(Registration, Evaluation, Authorisation
and Restriction of Chemicals, REACH),
EDs are covered by the authorisation
procedure for substances of very high
concern. Despite the joint efforts of many
organizations, there is still a need for agreed
Human Biomonitoring
test methods that can confirm whether or
not “identified candidates” (more than 500
compounds) are real EDs. As stated also
by the American EPA (Environmental
Protection Agency), validated methods
able to evaluate specific effects of EDs
are still being developed (22). Remarkably,
studies on environmental/occupational
exposure to EDs and reproductive risks
have been recently carried out, aiming at
dissecting the impact of these chemicals
on fertility. Exposure in utero at critical
developmental periods may modify the
normal path of reproductive and genitourinary development (23,24), and induce
hypospadia that is the most frequent
genital malformations in the male newborn
and results from an abnormal penile and
urethral development (25).
HBM fits with the scope of translational
research given that the expected results
include the validation of conventional/
new effect and exposure biomarkers,
the elucidation of the biomolecular
mechanisms of action of selected toxicants,
the identification of genetic susceptibility
markers in the Italian population, and the
definition of the criteria leading to the
evaluation of real exposure to toxicants.
In the early 1960s, powerful analytical
techniques allowed to measure very low
concentrations of chemical substances in
biological tissues caused by environmental
exposure. Due to the improvement of
these techniques essentially through
the effort of basic research, it is now
possible to detect very low concentrations
of agents (parts per trillion and parts
per quadrillion) with a high degree of
accuracy and precision. This general
consideration implies that measurement
procedures need a continuous validation
and that up-to-dating of basic knowledge
of biological effects of chemicals could
help in the development of new procedures
of risk assessment evaluation. The analysis
of benzene exposure, for instance, has
taken advantage from the old evidence
that trans,trans-muconic acid represents
the urinary metabolite, thus prompting
scientists to develop an ad hoc assay (26).
Many disciplines acquired a growing
and growing relevance in biomonitoring,
e.g. biophysics, whose contributions have
gone well beyond the mere application of
physical techniques to the study of living
systems. Biophysics plays a crucial role
in the development of new methodologies
and establishes closer links with other
frontier areas of the biological and medical
sciences (structure-function relations in
biological molecules, molecular biology,
evolution has widened the range of skills
required by the individual researcher and
has increased the need for teams with
diversified specializations.
Given that biomonitoring is not confined
to the exposure to toxicants but covers also
the identification of effect and susceptibility
markers, the final goal of HBM is to define a
toxicogenomic approach, considered as an
integration of genomics (transcriptomics,
proteomics and metabolomics) and
toxicology. This scientific field investigates
how the genome is involved in responses
to environmental stressors and toxicants.
It combines studies of mRNA expression,
cell and tissue-wide protein expression
and metabolomics, to understand the
role of gene-environment interactions in
disease. One of the important aspects of
toxicogenomic research is the development
and application of bioinformatics tools and
databases in order to facilitate the analysis,
mining, visualizing and sharing of the
vast amount of biological information
CNR Environment and Health Inter-departmental Project
being generated in this field. This rapidly
growing area promises to have a large
impact on many other scientific and
medical disciplines as scientists could
now generate complete descriptions of
how components of biological systems
work together in response to various
stresses, drugs, or toxicants. Of course,
this approach requires the joint effort of a
panel of experts, who accumulated a solid
experience through basic research, and can
transfer it to health applications. This is
the case of the teams involved in the PIASHBM project, which are characterized by
complementary expertise and capability to
cope basic and translational research.
3.1 CNR Institutes
The coordination of the HBM network
has been entrusted to the Institute of
Molecular Genetics (CNR-IGM, Pavia).
CNR-IGM research activity covers a
wide range of biological, biochemical
and genetic topics, and ensures the
participation of most CNR-IGM scientists
to the HBM project. This feature, i.e. the
potential commitment of a whole CNR
Institute to the Environment and Health
Inter-departmental project PIAS, has
rendered CNR-IGM especially suitable for
the coordination of the HBM group. The
HBM coordinator (Giuseppe Biamonti,
the present Director of CNR-IGM, then
replaced by A. Ivana Scovassi) was in
charge to identify CNR Institutes with the
expertise required to join CNR-IGM in
order to establish a PIAS-HBM network
based mainly on the analysis of the effects
of EDs (see below).
CNR-IGM is mainly devoted to basic
research on the control of cell proliferation,
DNA replication and apoptosis in
human cells; viral replication; analysis
of hereditary genetic disorders with
defects in DNA damage repair pathways,
chromosome X-linked diseases and
muscular dystrophies; post-transcriptional
regulation of gene expression during cell
response to stress treatments and tumor
progression, and analysis of the genetic
structure of human populations. Longlasting interactions and collaborations
within the CNR-IGM team members
are attested by joint peer-reviewed
publications. The scientific excellence
is completed by a qualified translational
research in collaboration with SMEs,
which originated a number of patents.
CNR-IGM researchers are active in
the development of new techniques,
protocols and instruments as well as in the
identification and characterization of new
compounds with therapeutic properties.
Considerable effort is put in the training of
undergraduate, graduate and post-doctoral
students. A group at IGM was recently
investigating the biological effects of toxic
metals and demonstrated the activation of
stress response mechanisms after cadmium
administration (27-29).
The Institute of Biophysics (CNR-IBF,
Genoa) covers a wide range of research
fields, sharing as a common feature the
application of typical methodologies and
techniques of the physical sciences to
develop interdisciplinary approaches to
the study of the structure and functions
of biological systems. A relevant interest
concerns physico-chemical investigations
of the impact of anthropic and nonanthropic environmental factors on
ecosystems. CNR-IBF devotes much effort
to the training of young people for research
in the fields of Biology and Biophysics, in
close collaboration with local universities.
The team involved in PIAS-HBM has a solid
Human Biomonitoring
research experience in electrophysiology
and ion channel biophysics in nervous
and endocrine culture cells investigated
by patch-recording and voltage-clamp
techniques, and intracellular calcium
dynamics, monitored by fluorescent
probes. The group studies heavy metal
accumulation and toxicity in mammalian
cells and modulation of neurotrasmittergated ion channels. The effect of acute and
chronic treatment with toxic metals (lead,
cadmium and nickel) on cell survival and
maturation of neurons in culture is currently
analyzed by functional and viability tests
and apoptosis/necrosis measurements (3036).
The Institute of Biomedicine and
Molecular Immunology (CNR-IBIM,
Palermo) is involved in research activity,
technological transfer and training in the
following areas: molecular, cellular and
morphological study of early embryo
development and mechanisms involved in
the differentiation and in the degenerative
mechanisms of eukaryotic cells; molecular
study of proteins involved in the allergic
reaction; synthesis and characterization
of bioactive molecules; pathophysiology
of the cardio-respiratory system, lung
diseases, systemic hypertension and
transplantation; bioeffects of magnetic
fields; epidemiology.
An CNR-IBIM team developed the
concept that sea urchin embryos as a new
friendly model for ecotoxicological studies
(37-41). Indeed, marine organisms are
highly sensitive to many environmental
pressures, and consequently, the analysis
of their bio-molecular responses to
different stress agents is very important
for the understanding of putative
repair mechanisms and for application
in environmental studies. Sea urchin
represents a simple though significant
model system where to test: 1) the impact on
the biology of development in association
with gene expression on embryos, and 2)
the effects on gene expression and DNA
damage on adult immuno-competent cells,
which are contained in the coelomic cavity
of the adult sea urchin, generically called
coelomocytes, studied since many decades,
but only recently used as bio-indicators of
stress. Due to the capability to respond
to injuries, host invasion and cytotoxic
agents, coelomocytes have been regarded
as the immune effectors of the sea urchin.
Another CNR-IBIM group investigates
the impact of environmental substances
both on hormone metabolism and nervous
system, focusing on the identification
of molecular mechanisms at the basis of
neurodegenerative diseases, included some
retinopathies. The group carries out in
vitro studies to elucidate the effect of target
chemicals on neurosteroidogenic pathways
and the interactions of these chemicals
with ER and AR receptors. On the other
hand, in vivo analyses using animal models
(mice) could be helpful in elucidating the
effects in age-related neurodegenerative
disorders. RT-PCR, receptor binding assay,
recombinant mammalian and yeast cell
based transcription assay, western blot,
immunocitochemical and histochemical,
proteomic and functional genomics are
currently used.
The Institute of Clinical Physiology
(CNR-IFC, Pisa), the largest biomedical
institute of the CNR’s Department of
Medicine, whose mission is “Innovation for
better patient care”, is involved in the study
of systemic, neuroendocrine and metabolic
factors implicated in many diseases.
Molecular medicine, clinical biology, and
clinical biochemistry are devoted to the
study of experimental physiology, and
CNR Environment and Health Inter-departmental Project
to the relevant diagnosis and treatment.
CNR-IFC is a leader institute in the field of
clinical and environmental epidemiology,
population registers, and research on health
services. CNR-IFC researchers established
a network of scientific collaborations with
many Italian and international Institutions.
CNR-IFC was the first public body in Italy
to achieve the status of pharmaceutical
developer (“Officina farmaceutica”) with
its own production site for injectable sterile
radiodrugs with Good Medical Practice
certification. This achievement was made
possible through close collaboration with a
leader Company in the field of biomedical
technology and diagnostic imaging, which
signed a contract for the production and
distribution of radiodrugs for diagnosis in
Positron Emission Tomography.
The Epidemiology unit works on
epidemiological surveillance, air pollution
and health, waste and health and is involved
in the national strategic programme
“Environment and Health”, funded by the
Ministry of Health and coordinated by
ISS. Human biomonitoring represents the
priority research area of this unit (42,43).
IFC has a long-standing experience in
the field of surveillance and research on
congenital malformations, as coordinator
of the EUROCAT and ICBDSR “Tuscany
Registry of Birth Defects”. To investigate
the correlation between EDs exposure and
reproductive dysfunctions, malformations
such as hypospadia or cryptorchidism,
reduced sperm counts, testicular cancer
and endometriosis, are studied (44).
3.2 External collaborations
Each CNR team collaborates with a number
of Italian and international Institutions,
including Universities, the Scientific
Institute for Research, Hospitaization
and Health Care (IRCCS), the National
Institute of Health (ISS), the World Health
Organization (WHO) and SMEs operating
in the biomedical/biotechnology field.
To increase the competences of the CNR
Biomonitoring team, external collaborators,
i.e. Salvatore Maugeri Foundation (FSM,
Pavia), Perrino Hospital (Brindisi) and
National Institute for Occupational Safety
and Prevention (ISPESL, Rome) have been
added to the CNR network on the basis of
their pre-existing cooperations with CNR
Institutes and taking into account their
complementary expertise.
FSM is a leading institution in the field of
the occupational health and prevention,
including biological and environmental
monitoring of exposure, reference values
setting and prevention of occupational
risks. The prevention of occupational risks
activity is supported by the Laboratory
for Environmental and Toxicological
Testing, which has been working for
decades on environmental monitoring,
with respect to occupational exposure to
both organic and inorganic substances
(risk assessing and sampling techniques),
and on the development of new biomarkers
and analytical techniques aiming at
the evaluation of occupational and
environmental exposure to xenobiotics.
This laboratory is fully equipped to
develop and validate analytical methods
for the determination of trace elements
(ICP-MS), organochlorinated compounds
such as PCB and DDTs (HRGC-MS),
phthalate metabolites (HPLC-MS/MS),
and other emerging substances in biological
and environmental matrices, including
foodstuffs. The Unit has recently validated
reliable methods for the determination of
EDs in biological fluids and in foodstuffs
ISPESL is a technical-scientific body in the
National Health Service and reports to the
Ministry of Health as regards all aspects of
Human Biomonitoring
Table 1: PIAS Human biomonitoring network.
Group leader
Patrizia Guarneri
Impact of EDs on steroid hormone metabolism
and neurodegenerative conditions
Valeria Matranga
IBF-CNR, Genova
(Materiali e Dispositivi)
Validation of biosensor methods for the analysis
of the effects of stress conditions on marine
Carla Marchetti
Gianfranco Prestipino
Identification of the mechanisms of action of
toxic metals in mammalian nervous cells
Anna Pierini
Fabrizio Bianchi
Epidemiological studies on the impact of
chemicals on human health
IGM-CNR, Pavia
Scienze della Vita)
Ivana Scovassi
Identification of effect biomarkers (in vitro and
in vivo approaches)
Fondazione Salvatore Maugeri,
Claudio Minoia
Biomonitoring of toxicants and their metabolites
in biological fluids from exposed people; total
diet studies
Ospedale Perrino, Brindisi
Giuseppe Latini
Biomonitoring of phthaltes: impact on infants
and human fertility
Elena Sturchio
Model systems: C. elegans; DNA biosensors;
miRNA platform
IBIM-CNR, Palermo
Scienze della Vita)
occupational safety, health and prevention.
Among the various activities carried out by
ISPESL, of interest are research, analysis,
experimentation and drafting of criteria
and methodologies for the prevention
of accidents and professional diseases,
identification of safety criteria, prevention
against chemical, physical and biological
exposure risks at work, standardization
of test to evaluate occupational safety and
health risk assessment.
The ISPESL Department of Production
Plants and Anthropic Settlements (DIPIA),
carries out research, experimentation,
consultation, assistance to the enterprises,
proposal of rules, laboratory controls,
standardization of methods and procedures
of evaluation, analysis of the systems for
purposes of safety and environmental
compatibility connected to the interaction
between the production premises and
the external environment. DIPIA is also
concerned with the complex problems
arising from biotechnologies, particularly
with the safety and risk assessment of
contained use of genetically modified
microorganisms and the evaluation of
genetically modified organisms safety
and their traceability in food and feed
products. DIPIA staff carries out many
projects supported by Ministry of Health
on eco-genotoxicity studies, biomonitoring
of polluted sites with toxicity testing, and
molecular analysis; it focuses on indicators
to detect the state of environmental health
after a release of pollutants from “fallout”. To develop potential biomarkers of
susceptibility, toxicity tests and plant and
animal models are employed, including the
nematode C. elegans, where toxicant effects
on phenotype, reproduction, apoptosis and
micro RNA profiles are tested. About this
CNR Environment and Health Inter-departmental Project
latter point, the analysis of miRNAs for
the detection of early indicators in various
diseases is a distinctive feature of the
PIAS-HBM team.
The Director of the Neonatology unit at the
Perrino Hospital (Brindisi) has recently
joined the EU pool of experts of risk
assessment. His main scientific interest
is the study of the effect of phthalates,
to whom general population is exposed
through consumer products, as well as
diet and medical treatments. In fact,
animal studies showing the existence of an
association between some phthalates and
testicular toxicity have generated public
and scientific concern about the potential
adverse effects of environmental changes
on male reproductive health. In addition,
prenatal exposure to phthalates seems
to play a relevant role in determining
these adverse effects given that human
exposure has been demonstrated to
begin during the intrauterine life. A link
between antenatal phthalate exposure and
abnormal fetal development exists, thus
justifying the need of therapeutic tools to
fight the adverse effect of this exposure.
Numerous maternal lipophilic compounds
are eliminated into milk during lactation,
their concentrations reflecting fetal in
utero exposure. The reported effects of the
exposure to phthalates through breast milk
in infants confirm that human milk may
represent an additional potential source
of phthalate exposure in a population at
increased risk (49-51). This research is made
in collaboration with CNR Institutes.
The members of the “PIAS-HBM network”
are listed in Table 1, which reports also
the specific (and unique) expertise of each
The CNR-IBF team has depicted the
molecular mechanisms of nickel and lead
toxicity in neural cells. Focusing on the
inhibition of N-methyl-D-aspartate receptor
(NR) channel in a voltage-dependent
manner, the group recently identified
specific heavy metal interaction with NR
channels (30) and defined the relevance of
NR composition in modulating the effect
of toxic metals (31). The survey of lead
effects represented the first structural work
addressing the location of lead interaction
site on NMDA receptor channel, thus
providing original electrophysiological
data (30); the new results obtained for
nickel allowed the characterization of
its involvement in synaptic currents and
transmission (31).
At CNR-IBIM, it has been established
the suitability of sea urchin as a sentinel
organism for the assessment of the
macro-zoobenthos health state in biomonitoring programmes. A recent survey
of sea metal contamination around
Pianosa and Caprara Islands revealed that
sea urchin coelomocytes might be used as
biosensors of environmental stress (52,53).
This observation further supports the
evidence that sea urchin as well as marine
invertebrates are useful as bioindicators of
environmental stress.
The SEBIOMAG project represents a good
example of an integrated approach to HBM.
SEBIOMAG (Studio Epidemiologico
Biomonitoraggio Area di Gela) was
promoted by World Health Organization,
(CNR-IFC), and carried out in collaboration
with the Laboratory for Environmental and
Toxicological Testing at FSM (Pavia). The
project was conceived as the biomonitoring
Human Biomonitoring
Table 2: Published monographs of relevant EDs.
HBM teams
Bisphenol A
Minoia C, Leoni E, Turci R, Signorini S,
Moccaldi A, Imbriani M
[55] G Ital Med Lav Erg
2008; 30:214-24.
Minoia C, Leoni E, Sottani C, Biamonti G,
Signorini S, Imbriani M
[56] G Ital Med Lav Erg
2008; 30:309-23.
Sturchio E, Minoia, Zanellato M, Masotti A,
Leoni E, Sottani C, Biamonti G, Ronchi A,
Casorri L, Signorini S, Imbriani M
[57] G Ital Med Lav Erg
2009; 31:5-32.
[58] G Ital Med Lav Erg
2009.; 31:325-70.
Turci R, Minoia C, Leoni E, Sturchio E, Boccia P,
Meconi C, Zanellato M, Signorini S, Benzoni I,
Mantovani A, La Rocca C, Bianchi F, Imbriani M
of the exposure of target population living
in Gela (Sicily). The study consisted of the
evaluation of toxicant levels in biological
fluids, focusing on trace elements
(antimony, arsenic, beryllium, cadmium,
mercury, lead, copper, selenium, thallium,
vanadium) and organochlorine compounds
(PCB, aldrin, dieldrin, DDT, chloroesane,
chlorobenzene). Trace elements and
organochlorine compounds were measured
by inductively coupled plasmadynamic
reaction cell-mass spectrometry (ICPDRC-MS) and gas chromatography
coupled to mass spectrometry (GCMS), respectively. The target population
consisted in 184 subjects aged 20-44 years
living in Gela (116), Niscemi (39) and
Butera (29). The analysis revealed that
the levels for organochlorine compounds,
antimony, selenium, thallium, beryllium
and vanadium were comparable to control
population. The most relevant finding was
the peculiar profile of widespread exposure
to arsenic recorded in blood, plasma and
urine samples, where the values were
higher than reference values of nonexposed subjects. In conclusion, the results
underlined a widespread exposure to
arsenic, and recommended the evaluation
of different forms of arsenic (inorganic
arsenic, arsenobetaine, arsenocholine) to
better understand the potential sources of
exposure (environmental, seafood intake)
in this crucial area. The results of this
study have been presented in Gela, on July
16, 2009, at the SEBIOMAG meeting.
Remarkably, this analysis represents
an added value to a previous survey
conducted on the same geographical area
and providing environmental data about
contaminants present in water, earth and
air. These data are reported in the special
issue of the journal Epidemiology &
Prevention devoted to “Environment and
health in Gela (Sicily): present knowledge
and prospects for future studies” (54). The
joint effort to evaluate the environmental
contamination of an industrial area,
and the consequent exposure of the
general population, is an example of an
efficient strategy to combine clinical and
research competences for improving the
experimental approaches useful to depict
the effects of environmental risk on human
CNR Environment and Health Inter-departmental Project
Of note, some members of the network
cooperate for preparing toxicological
information profiles of specific chemicals.
Based on a huge amount of literature data,
these reviews provide relevant scientific
information actually on EDs, including
bisphenol A, perfluoroalkyl agents (i.e.
PFOS/PFOA), trace elements (i.e. arsenic)
and dioxins (i.e. PCDDs) (55-58). The
papers, listed in Table 2, are published on the
“Giornale Italiano di Medicina del Lavoro
e Ergonomia”, that is the official Journal
of FSM. Each paper includes general
aspects of exposure source, chemical
and physical properties, metabolism,
toxicological and carcinogenic potential,
food intake and diet exposure, mechanism
of action, genetic susceptibility, analytical
procedures and general population levels.
This body of information represents a
very useful tool not only for clinicians but
also for researchers in the environmental
and occupational context. In fact, even
if extensive toxicity data for a chemical
are available, they are almost always in
a form that is difficult to combine with
biomonitoring-generated exposure values
to assess risk (15).
An example of scientific literature that aims
at making attractive and understandable
to the general public a so complex topic
as Environment&Health is represented
by the recent book “Ambiente e salute:
una relazione a rischio. Riflessioni tra
etica, epidemiologia e comunicazione”
by the scientists from the CNR-IFC
Epidemiology unit (59). The book uses
a number of case studies to define the
concept of epidemiology, population
complexity, scientific communication, and
future perspectives in risk assessment.
The hot topic “Environment&Health” is
discussed at different levels, that is ethics,
epidemiology and communication also in
a recent publication (60).
In the last years there has been a call
for increased epidemiological and
experimental research to substantiate
the disruptive effects of environmental
chemicals in humans. This is currently
recognized as an important issue of concern
in the protection of human environmental
Human biomonitoring is an important tool
to evaluate the internal exposure through
the environment and to provide early
warning indicators for possible long-term
adverse effects. Biomonitoring has two
main goals: i) determination of the levels of
toxicants in biological fluids from a general
population; ii) search for new exposure,
effect and susceptibility biomarkers.
The here described PIAS-HBM network
combines solid clinical expertise with
advanced research work. With an adequate
funding, it could act in a coordinated way
to address the following crucial points:
1. To handle the Environment&Health
problem with a multidisciplinary
approach, combining medical tools
with a biological approach based on
biochemistry, biophysics, cell and
molecular biology, bioinformatics,
molecular genetics and genomics. This
goal could be achieved through a strict
cooperation between the different
members of the network, sharing the
respective competences and working
in tight association.
2. To validate conventional/new exposure
biomarkers. The growing number of
chemicals potentially toxic for human
health implies a continuous effort to
develop new specific assays and to
Human Biomonitoring
validate them; in parallel, conventional
procedures have to be periodically
checked for their efficacy by different
laboratories. As for exposure markers,
it could be of interest to share biological
samples collected at Hospitals and
IRCCS in order to increase the
number of selected biomarkers. As
a new perspective, it could be useful
to evaluate the real exposure of the
general population through the diet,
which is considered to be the main
source of body burden of several
contaminants. This approach, based
on a validated protocol that estimates
the dietary intake of compounds,
could contribute to face the lack of
data on the real exposure to several
contaminants and to develop ad hoc
protocols to measure their levels in
food. Quantification of the risk by
the ingestion of pollutants in food is
complex and depends on many factors
(species, diet composition, duration
of exposure, efficiency of pollutant
absorption, subsequent metabolism,
sensitivity of target organs and stage
of development). While the effects
of high doses of single chemicals are
proven, dietary exposure generally
involves prolonged, low-level exposure
to a large number of compounds,
each of which has different chemical
characteristics, different biological
effects and is present at varying
3. To validate conventional/new effect
biomarkers. The search for effect
biomarkers implies a panel of
approaches, spanning from cellular
to molecular biology, focusing on
the effects of chemicals in terms of
cell proliferation, cell death, signal
transduction, and neurodegeneration.
The PIAS-HBM team possesses the
ability to focus on the dissection of the
biochemical steps of basic biological
processes, such as DNA replication
and repair, transcription, translation
and post-translation. In vivo assays
on cell lines of different origin (e.g.
neuroendocrines, neurals, fibroblasts,
keratinocytes, transformed) may
allow the identification of the effect
of chemicals on different cellular
(adhesion, cell cycle, proliferation
and death, motility) and biochemical
(DNA damage, electrophysiology,
metabolism, neurotoxicity, ionic
transport, replication and repair,
oxidative stress) parameters. Also the
analysis of the impact of chemicals
on different classes of receptors (e.g.
ER alpha/beta, AR, PR, GR, ThRs,
retinoic acid, aryl-AhR, pregnaneX-R) is appealing. The measurement
of the activity of a number of reference
enzymes (e.g. acetylase, 5-alfa
reductase, aromatase, kinase, DNA
polymerase, phosphorylase, ligase,
nuclease, PARP, protease, telomerase,
topoisomerase, reverse transcriptase,
sulfatase, sulfotransferase) represents
an original tool to elucidate the
biomolecular mechanisms of action
of selected toxicants and to identify
new effect biomarkers to be tested
in the population through validated
4. To identify genetic susceptibility
markers in the Italian population. The
search for susceptibility markers in the
Italian population is a priority issue.
In addition to cytogenetic and genetic
assays, new molecular biology tools
such as microRNA and epigenetic
regulation of gene expression are
useful to obtain predictive markers
of noxious effects of pollutants
CNR Environment and Health Inter-departmental Project
correlated to the genetic background
of individuals. The final goal is to
identify toxicogenomic markers in the
Italian population.
In addition to the above action plan, which
is strictly based on the specific know-how
of the single teams, other hints emphasize
the power of our network:
a) Human biomonitoring is directed to
human beings. However, basic research
on organisms other than humans could
speed up the process of biomarker
identification and limit the use of
human material. The investigations at
ISPESL aiming at validating C. elegans
as model organism for the analysis
of the impact of toxicants on living
organisms could be very useful. In fact,
the fast growth and reproduction time
of this animal as well as the exhaustive
knowledge of this genome renders C.
elegans an extremely easy model. As
underlined in the 2002 Nobel prize
award (61), the identification of key
genes regulating organ development
and programmed cell death in C.
elegans was instrumental to the
knowledge that corresponding genes
exist in higher species, including
man. Similarly, the body of evidence
accumulated at CNR-IBIM on the use
of sea urchin as biosensor could have
more applications than the actual ones.
Since the publication of the sequence
of this marine invertebrate organism
(62), new experimental procedures
have been developed to test the effect
of chemicals both on whole marine
organisms and on coelomocytes. This
is a new and original tool.
b) It is widely agreed that exposure in utero
to toxicants may modify the normal
path of reproductive development
and cause genital malformations
(hypospadia, cryptorchidism). In the
same scientific area, it has been shown
that prenatal exposure to pollutants
and/or the exposure through the
maternal milk have adverse effect
of infant health. The incidence
of congenital malformations may
represent an early biological indicator
for environmental and/or occupational
exposure to contaminants. The search
for factors affecting the maternal-fetal
environment is currently addressed at
CNR-IFC and Perrino Hospital.
c) A correlation between neurological
disorders and environmental risk
has been found. The expertise of
CNR-IBF and IBIM in the field of the
study of the effects of pollutants on the
nervous system strongly supports an
in-depth examination of the problem
of genetic and environmental factors
that modulate the occurrence of such
d) There is an increasing need for
epidemiology studies, to develop
a strategy of protection of human
environmental health. Within our
PIAS-HBM network, the expertise
of CNR-IFC guarantees a correct
approach to the problem.
e) The compilation of reviews on the
toxicological profile of pollutants
is extremely useful to the scientific
community. Further work is required
to increase the number of considered
chemicals; to this purpose, the
involvement of all the members of the
PIAS-HBM team is desirable. This
activity will facilitate clinicians and
In conclusion, it is clear that in order
to investigate the relationship between
environment and health, the most
Human Biomonitoring
important prerequisite is the availability
of an appropriate panel of biomarkers
as indicators of individual exposure to
environmental chemicals. This attempt
requires a cross-interaction among
toxicology, cellular and molecular
biology in order to identify a potent body
of biomarkers of exposure, effect and
susceptibility. In this respect, our PIASHBM network fits with the purposes of
modern HBM, which has expanded beyond
its origin in occupational medicine to cover
a wide variety of diagnostic procedures and
assessments of environmental pollution,
leading to the identification of potentially
hazardous exposure before adverse health
effects appear and to establish exposure
limits for minimizing the likelihood
of significant health risk methods. The
network we established can be enlarged
to other Italian teams providing additional
competences, thus ensuring a multifaceted
research approach, possibly under the
aegis of CNR.
The research of the groups belonging to the
PIAS-HBM network is funded by different
granting agencies. CNR-IGM activity is
globally supported by several international
grants from the European Framework
Programmes, as well as from private and
public national agencies (Italian Ministry
of Health, MIUR, ISS, AIRC, Telethon,
Cariplo, Regione Lombardia). Special
funds from CNR have been allowed to
CNR-IGM to coordinate the HBM group
within the frame of the PIAS project.
The granted sum has covered a one-year
research fellow, and the organization of
and attendance to the PIAS-CNR Seminar
“Endocrine disruptors for EnvironmentHealth evaluation” (Rome, March 16, 2009),
to the kick-off HBM meeting (Rome, March
17, 2009), and to the PIAS-CNR Workshop
“Advancement and perspectives” held in
Rome on June 18, 2009. FSM is mainly
granted by Italian Ministry of Health.
On the behalf of Research Agreements,
CNR-IBF carries out several projects in
collaboration with the University of Science
and Technology of China, a group in
Poland, and the Institute de Biotecnologia/
UNAM Cuernavaca, Mexico. CNR-IBIM
activity in the biomonitoring domain is
supported by various grants, including
the Bilateral Italy (CNR)-Japan (JSPS)
Seminar on “Physical and Chemical
Impacts on Marine Organisms”. CNR-IFC
has institutional liaisons for exploiting
innovation, with Regione Toscana, local
industry, ISS and WHO. Among the
financed Environment&Health projects, of
interest are the following: “Epidemiological
study of biomonitoring” in Campania
Region (SEBIOREC) funded by ISS;
“Epidemiological study of biomonitoring
in Gela Area” (SEBIOMAG) funded
by WHO. ISPESL received a financial
support from Ministry of Health to carry
out projects in the field of “Prediction,
prevention and protection of human health”,
and “Development of advanced biosensors
for environmental monitoring”.
That each member of the PIAS-HBM
team has been/is granted to work in the
risk assessment field is an evidence of the
qualification of the scientists belonging
to the network. However, to establish a
coordinated and multidisciplinary work
strategy, a global grant from a public
agency, covering the expenses of an
integrated project, is absolutely required.
We would like to thank the colleagues
who shared with us their expertise in the
Environment&Health field and accepted
to participate in the PIAS-HBM team.
In particular, we kindly acknowledge the
active collaboration of Patrizia Guarneri,
Giuseppe Latini, Carla Marchetti, Valeria
CNR Environment and Health Inter-departmental Project
Matranga, Claudio Minoia, Anna Pierini,
Gianfranco Prestipino and Elena Sturchio.
We are indebted to the PIAS coordinator,
Fabrizio Bianchi, for his continuous and
invaluable support.
Keywords biomarkers, biomonitoring, EDs,
epidemiology, pollutants.
DeCaprio AP. Biomarkers: coming of
age for environmental health and risk
assessment. Environ Sci Tech 1997;
2. Available
3. Metcalf SW, Orloff KG. Biomarkers of
exposure in community setting. J Toxicol
Environ Health 2004; 67:715-26.
4. Budnik LT, Baur X. The assessment of
environmental and occupational exposure
to hazardous substances by biomonitoring.
Dtsch Arztebl Int 2009; 106:91-7.
5. Angerer J, Ewers U, Wilhelm M. Human
biomonitoring: state of the art. Int J Hyg
Environ Health 2007; 210:201-28.
6. Available from: URL:http://www.invs.
7. Colborn T, vom Saal FS, Soto AM.
Developmental effects of endocrinedisrupting chemicals in wildlife and
humans. Env Health Perspect 1993;
8. Crisp TM, Clegg ED, Cooper RL et al.
Environmental endocrine disruption:
an effects assessment and analysis. Env
Health Perspect 1998; 106 Suppl 1:11-56.
9. Available from: URL:
10. Available from: URL:
11. Available from: URL:
12. Casteleyn L, Tongelen BV, Fatima Reis M,
Polcher A, Joas R. Human biomonitoring:
Towards more integrated approaches in
Europe. Int J Hyg Environ Health 2007;
210: 199-200.
Available from: URL:http://eur-lex.
eu / LexUr iSer v/ LexUr iSer v.
Available from: URL: http://ec.europa.
eu /resea rch /envi ron ment /pdf / hbmconference-highlights_conclusions_
Available from: URL:
Available from: URL:http://www.euro.
Available from: URL:http://www.eea.
Available from: URL:
Järup L. Hazards of heavy metal
contamination. Br Med Bull 2003; 68:16782.
Hotchkiss AK, Rider CV, Blystone CR
et al. Fifteen years after “Wingspread”environmental endocrine disrupters and
human and wildlife health: where we are
today and where we need to go. Toxicol
Sci 2008; 105:235-59.
Communities. Community strategy for
endocrine disruptors. COM 1999; 0706:131.
Available from: URL:http://www.epa.
Foster WG, Neal MS, Han MS, Dominguez
MM. Environmental contaminants and
human infertility: hypothesis or cause for
concern? J Toxicol Environ Health B Crit
Rev 2008; 11:162-76.
Small CM, DeCaro JJ, Terrell ML et al.
Maternal exposure to a brominated flame
retardant and genitourinary conditions
in male offspring. Env Health Perspect
2009; 117:1175-9.
Wang MH, Baskin LS. Endocrine
disruptors, genital development, and
hypospadias. J Androl 2008; 29:499-505.
Scherer G, Renner T, Megel M. Analysis
and evaluation of trans, trans-muconic
acid as a biomarker for benzene exposure.
J Chromatogr B Biomed Sci Appl 1998;
Human Biomonitoring
27. Chiodi I, Biggiogera M, Denegri M et
al. Structure and dynamics of hnRNPlabelled nuclear bodies induced by stress
treatments. J Cell Sci 2000; 113:4043-53.
28. Denegri M, Chiodi I, Corioni M,
Cobianchi F, Riva S, Biamonti G. Stressinduced nuclear bodies are sites of
accumulation of pre-mRNA processing
factors. Mol Biol Cell 2001; 12:3502-14.
29. Valgardsdottir R, Chiodi I, Giordano M
et al. Transcription of Satellite III noncoding RNAs is a general stress response
in human cells. Nucleic Acids Res 2008;
30. Gavazzo P, Guida P, Zanardi I, Marchetti
C. Molecular determinants of multiple
effects of Nickel on NMDA receptor
channels. Neurotox Res 2009; 15:38-48.
31. Gavazzo P, Zanardi I, BaranowskaBosiacka I, Marchetti C. Molecular
determinants of Pb2+ interaction with
NMDA receptor channels. Neurochem
Int 2008; 52:329-37.
32. Gavazzo P, Mazzolini M, Tedesco M,
Marchetti C. Nickel differentially affects
NMDA receptor channels in developing
cultured rat neurons. Brain Res 2006;
33. Marchetti C, Gavazzo P. NMDA receptors
as targets of heavy metal interaction and
toxicity. Neurotox Res 2005; 8:245-58.
34. Cupello A, Esposito A, Marchetti
C, Pellistri F, Robello M. Calcium
accumulation in neurites and cell bodies
of rat cerebellar granule cells in culture:
effects on GABAA receptors function.
Amino Acids 2005; 28:177-82.
35. Prestipino G, Corzo G, Romeo S et al.
Scorpion toxins that block transient
currents (IA) of rat cerebellum granular
cells. Toxicol Letters 2009; 187:1-9.
36. Romeo S, Corzo G, Vasile A, Satake H,
Prestipino G, Possani LD. A positive
charge at the N-terminal segment of
Discrepin increases the blocking effect
of K+ channels responsible for IA currents
in cerebellum granular cells. Biochim
Biophys Acta 2008; 1780:750-5.
37. Russo R, Bonaventura R, Zito F et al. Stress
to cadmium monitored by metallothionein
gene induction in Paracentrotus lividus
embryos. Cell Stress Chaperones 2003;
Bonaventura R, Poma V, Costa C,
Matranga V. UVB prevents skeleton
growth and stimulates the expression of
stress markers in sea urchin embryos.
Biophys Biochem Res Comm 2005;
Schröder HC, Di Bella G, Janipour N et al.
DNA damage and developmental defects
after exposure to UV and heavy metals
in sea urchin cells and embryos compared
to other invertebrates. Prog Mol Subcell
Biol 2005; 39:111-37.
Pinsino A, Della Torre C, Sammarini
V, Bonaventura R, Amato E, Matranga
V. Sea urchin coelomocytes as a novel
cellular biosensor of environmental stress:
a field study in the Tremiti Island Marine
Protected Area, Southern Adriatic Sea,
Italy. Cell Biol Toxicol 2008; 24:541-52.
Yokota Y, Matranga V. Physical and
chemical impacts on marine organisms.
Marine Biol 2006; 149:1-5.
Bianchi F. From descriptive studies
towards epidemiologic surveillance.
Epidemiol Prev 2009; 33:127-32.
Linzalone N, Bianchi F. Human
biomonitoring to define occupational
exposure and health risks in waste
incinerator plants. Int J Environment and
Health 2009; 3: 87-105.
Dolk H, Vrijheid M, Scott JES et al.
Towards the Effective Surveillance of
Hypospadias. Environ Health Perspect
2004; 112:398-402.
Turci R, Balducci C, Brambilla
G et al. A simple and fast method
for the determination of selected
organohalogenated compounds in serum
samples from the general population.
Toxicol Lett. 2010; 192:66-71.
Turconi G, Minoia C, Ronchi A, Roggi C.
Dietary exposure estimates of twenty-one
trace elements from a Total Diet Study
carried out in Pavia, Northern Italy. Br J
Nutr 2009; 101:1200-8.
Turci R, Finozzi E, Catenacci G,
Marinaccio A, Balducci C, Minoia C.
CNR Environment and Health Inter-departmental Project
Reference values of coplanar and noncoplanar PCBs in serum samples from
two italian population groups. Toxicol
Lett 2006; 162:250-5.
Turci R, Turconi G, Comizzoli S, Roggi
C, Minoia C. Assessment of dietary intake
of polychlorinated biphenyls from a total
diet study conducted in Pavia, Northern
Italy. Food Addit Contam 2006; 23:91938.
Latini G, Del Vecchio A, Massaro M,
Verrotti A, De Felice C. In utero exposure
to phthalates and fetal development. Curr
Med Chem 2006; 13:2527-34.
Latini G, Del Vecchio A, Massaro
M, Verrotti A, De Felice C. Phthalate
exposure and male infertility. Toxicology
2006; 226:90-8.
Latini G, Wittassek M, Del Vecchio
A, Presta G, De Felice C, Angerer J.
Lactational exposure to phthalates
in Southern Italy. Environ Int 2009;
Pinsino A, Della Torre C, Sammarini
V, Bonaventura R, Amato E, Matranga
V. Sea urchin coelomocytes as a novel
cellular biosensor of environmental
stress: a field study in the Tremiti Island
Marine Protected Area Southern Adriatic
Sea, Italy. Cell Biol Toxicol 2008; 24:54152.
Matranga V, Yokota Y. Responses
of marine organisms to physical and
chemical impacts. Cell Biol Toxicol 2008;
Musmeci L, Bianchi F, Carere M, Cori L.
Environment and health in Gela (Sicily):
present knowledge and prospects for
future studies. Epidemiol Prev 2009; 33
Suppl 1:1-159.
Minoia C, Leoni E, Turci R, Signorini S,
Moccaldi A, Imbriani M. Bisphenol A. G
Ital Med Lav Ergon 2008; 30:214-24.
Minoia C, Leoni E, Sottani C, Biamonti
G, Signorini S, Imbriani M. Interferenti
Endocrini: Schede Monografiche 2. PFOS
e PFOA. G Ital Med Lav Ergon 2008;
Sturchio E, Minoia C, Zanellato M
et al. Interferenti Endocrini: Schede
monografiche 3. Arsenico. G Ital Med
Lav Ergon 2009; 31:5-32.
Turci R, Minoia C, Leoni E et al.
monografiche 4. PCDD:policlorodibenzop-diossine. G Ital Med Lav Ergon 2009;
Battaglia F, Bianchi F, Cori L. Ambiente e
salute: una relazione a rischio. Riflessioni
tra etica, epidemiologia e comunicazione.
Rome: Il Pensiero Scientifico Editore;
URL:http:// prizes/medicine/
Sea Urchin Genome Sequencing
Consortium, Sodergren E, Weinstock
GM et al. The genome of the sea urchin
Strongylocentrotus purpuratus. Science
2006; 314:941-52.
Environmental Health Surveillance Systems
E.A.L. Gianicoloa, A. Brunia, M. Serinellib
a. CNR, Institute of Clinical Physiology (IFC), Lecce, Italy
b. Regional Environmental Protection Agency, Puglia, Bari
[email protected]
An integrated environmental health surveillance system is the systematic, ongoing collection and analysis
of information related to disease and environment, and its dissemination to individuals and institutions.
This type of system provides scientific evidence and tools for implementing and evaluating policies aimed
at preventing, controlling and protecting health and the environment. An integrated environmental health
surveillance system can be realized by setting up environmental and health indicators. Indicators are useful
for understanding the spatial and temporal trends of environmental parameters and related health effects,
both acute and chronic.
In recent years, much attention has been focused on surveillance systems, and, in particular, on developing
methods for combining and integrating information in order to better understand these phenomena. Several
analytical approaches have been proposed for classifying environmental and health indicators: Thacker’s
model; the DPSEEA framework (WHO) which represents an evolution of the PSR model (OECD) and the
DPSIR framework (European Environmental Agency).
As part of the Environment and Health Inter-departmental Project (PIAS-CNR), the Working Group on
Environmental Health Surveillance Systems has been tasked with the development of a protocol to be tested
in areas with different environmental risks, in order to monitor environment and health indicators and to
provide useful tools for primary prevention programs and communication. The goal is to select a set of
environmental and health indicators to be assessed against their utility and availability in time and space.
Environmental effects on health are
associated with many different factors:
environmental degradation, such as
air, water and soil pollution, and food
contamination; global environmental
problems, such as the reduction of
biodiversity, the degradation of the
ecosystem through deforestation, global
warming, ozone layer depletion and
contamination by persistent organic
chemicals, waste cycle mismanagement
and industrial disasters.
As part of the Environment and Health
Inter-departmental Project (PIAS-CNR),
the Working Group on Environmental
Health Surveillance Systems has been
charged with developing a protocol to be
tested in areas with different environmental
risks, in order to monitor environment and
health indicators and to provide useful
tools for primary prevention programs
and communication. The analysis of the
scientific literature on this subject, and the
study of the most advanced international
experiences are the basis on which a
protocol can be defined and tested in areas
with different degrees of environmental
degradation. In Italy, the Apulia region,
a territory with three areas at high risk of
environmental crisis (Brindisi, Taranto
and Manfredonia),
is an interesting
experimental site.
CNR Environment and Health Inter-departmental Project
The areas chosen to test the surveillance
system require epidemiological and
environmental characterization, that can
be performed using available statistical
information or conducting specific
These elements, based on a conceptual
model (Thacker, DPSEEA or DPSIR)
integrated with established international
experience and shared with local and
national stakeholders, are the platform on
which to build an information system that
can provide information on environmenthealth interaction and lead to preventive
and communication actions.
This chapter aims at providing a summary
of our knowledge on this subject, with
particular regard to several international
experiences that are generally considered
as more advanced.
Major conceptual models in the field of
environment and health indicators will be
2.1 Definitions and purpose
Public health surveillance is “the
ongoing systematic collection, analysis,
and interpretation of outcome-specific
data, closely integrated with the timely
dissemination of these data to those
responsible for preventing and controlling
disease or injury“ (1). The modern
definition of surveillance, which at the
beginning included only transmittable
diseases (2), currently refers to chronic
and acute diseases, reproductive health,
work, domestic and road accidents,
environmental and occupational risk, and
behavior (3).
In 1988, the Centers for Disease Control
and Prevention (CDC) of the United
States defined environmental health
surveillance as ”the ongoing, systematic
collection, analysis, and interpretation
of health data essential to the planning,
implementation, and evaluation of public
health practice, closely integrated with
the timely dissemination of these data to
those who need to know. The final link of
the surveillance chain is the application
of these data to prevention and control. A
surveillance system includes a functional
capacity for data collection, analysis, and
dissemination linked to public health
programs.” (4).
Most surveillance systems are usually
implemented in order to:
- Provide estimates on the size of a health
- Investigate emerging health problems
and epidemics;
- Document the distribution and diffusion
of health events on a given territory and
in specific populations;
- Provide the basis for epidemiological
research and clinical trials;
- Describe the natural history of a
- Monitor trends in risk factors related to
specific diseases;
- Identify changes in health practices;
- Monitor the spatial and temporal
variation of the occurrence of diseases
and risk groups;
- Evaluate programs for prevention and
disease control (1).
In recent years, attention has been
increasingly focused on the need to improve
environment and health monitoring
systems by developing methods designed
to combine information from different
information-systems and to support an
integrated knowledge of phenomena.
An environmental health surveillance
system must be able to assess, analyze and
disseminate the information necessary to
properly plan policy-makers‘ actions in
Environmental Health Surveillance Systems
Figure 1. the process by which an environmental agent produces an adverse effect and
the corresponding types of public health surveillance.
health care.
Environmental indicators (quality of the
environment, environmental contamination
and results of specific monitoring) and
health indicators (e.g, indicators of
morbidity, mortality and reproductive
health) which can be obtained by current
health information flows, from a pathology
registry or from specific surveys, are the
bases of an integrated environmental health
surveillance system.
Environmental and health indicators provide
a quantitative summary of the phenomenon
under study and are useful to understand
the spatial and temporal patterns of healthimpacting environmental parameters ,
acute and chronic health effects and social
and demographic factors.
2.2 Conceptual models
Several models have been proposed to
provide a conceptual synthesis of the
monitoring of environmental and health
problems. These include
- The Thacker model (5);
- DPSIR and DPSEEA model (6, 7).
They emphasize the role of social and
environmental macro-determinants and
consider exposure to be a central event in
environmental causes and in the occurrence
of disease. Therefore, exposure is the
key element in environmental and health
surveillance. In fact, epidemiological
surveillance systems have developed,
from disease surveillance to surveillance
of collective risk factors (8).
2.3 The Thacker model
The Thacker model (5) proposes three
different kinds of surveillance (Fig. 1):
- Hazard surveillance;
- Exposure surveillance;
- Outcome surveillance.
Thacker defines hazard surveillance as
the “assessment of the occurrence of,
distribution of, and the secular trends in
levels of hazards (toxic chemical agents,
physical agents, biomechanical stressors,
as well as biological agents) responsible
for disease and injury” (9).
CNR Environment and Health Inter-departmental Project
Figure 2. The DPSIR model
Data on risk factors can be derived from
the amount of hazardous agents produced,
sold, used or released, or from the
concentrations of these agents in various
environmental matrices (air, food, soil and
dust, water) (10).
Exposure surveillance is the monitoring
of individual members of the population
to assess the presence of an environmental
agent or its clinically unapparent (e.g,
subclinical or preclinical) effects. (5, 11).
The definition of target groups for
surveillance in areas with documented or
presumed environmental pressure is one of
the most critical points in the researcher’s
decision-making chain, since it depends
on many factors and considerations.
Actually, exposure is defined as the
relationship between the environment
(external factors) and the individual
(internal factors) as a result of inhalation,
ingestion, dermal contact, or via fetus or
The need to establish a relationship
between environmental monitoring and
health-related policies and actions led to the
addition of a fourth monitoring category
regarding the assessment of policy options
2.4 The DPSIR and DPSEEA
The methodology of the
Impacts, Responses) allows us to
organize environment-health indicators
implemented in an environmental health
surveillance system (12) (Fig. 2).
The DPSIR framework is a system to
analyze the Driving forces responsible
for change, the resulting environmental
Pressures on the State of the environment,
the Impact of changes on environmental
quality, and Society’s Response to these
In this model the determinants (or sources,
e.g. Agriculture, industry, transport,
settlements, animal husbandry, mining)
identify the factors influencing the
environmental conditions as sources
on which to act. They are useful to
identify relationships between the factors
responsible for pressure and the pressure
Pressures (e.g. emissions of pollutants,
waste, noise emission , vibration and
radiation) identify the direct effects
of increased human activities (i.e. the
variables responsible for the degradation)
and are useful to quantify the causes of
environmental change.
States (e.g. quality of air, water,
soil, vegetation, fauna, ecosystems,
landscape, physical agents, public health)
represent environmental quality and the
environmental resources that should be
protected. They are useful to evaluate
environmental conditions in terms of
degree of impairment.
Impact refers to the effects of a pressure:
they are major changes in the environment
compared to a state-based condition, taken
as a reference.
Responses (laws, plans, rules) are actions
taken to address the impact, and take
different forms depending on the level of
the model on which to act (e.g. demands
of structural determinants, interventions
prescriptive or technology, etc.) (13).
Environmental Health Surveillance Systems
Each of the areas identified above can be
summarised by using specific indicators, as
resulting from current environmental and
health data, environmental bio-monitoring
and biomarkers of human exposure.
The model comes from the general concept
when applied to specific environmental
areas such as environmental matrices that
define the real component within which
chemical, physical and biological agents act.
Matrices are generally identified as air,
water, soil, waste, physical agents, and
foods. To better address the effect of the
human exposure to environmental factors,
the World Health Organization (WHO)
extended DPSIR to the DPSEEA model
(Determinant, Pressures, State, Exposure,
Effect, Action).
The introduction of health effects
evaluation involves a refinement of the
DPSIR conceptual model, translating the
concept of impact into ”exposure“ and
”effect“ and the Responses into ”actions”.
In the DPSEEA model, according to
the classical epidemiological model,
Determinants and Pressures are recognized
as determinants of disease, distinguished in
individual and contextual determinants.
State, Exposure and Effect represent the
extent of the problem (respectively, in
terms of emissions, exposure and health
effects) (13).
The number of indicators relating to the
areas outlined above is very high. There
is abundant literature on the selection of
indicators useful to describe the state of
the environment and health (7, 14).
The indicators include both environmental
and health indicators for which the
environmental hazards and health effects
is already established.
Wills and Briggs (15) define two categories
of indicators:
- Health-related environmental indicators
- Environment-related health indicators
The first relates to environmental
conditions that suggest potential harmful
health effects; the latter relates to health
outcomes that suggest an environmental
cause or a contribution from environmental
In environmental impact assessment
studies, an indirect measure of the level of
exposure (e.g. concentrations of pollutants
or emissions) is used as an environmental
indicator (16).
In studies of health impact assessment
of environmental pollution, indicators
that describe health outcomes caused by
exposure to polluted matrices are used.
To determine which diseases are related
to the environment (e.g. infant mortality,
mortality from respiratory causes), it is
necessary to carry out studies on risk
assessment from exposure (16).
In general, indicators should have specific
- Validity,
representativeness of data;
- Availability of data, their systematic
measurement in time and space (not
separated from their representation);
- Usefulness, i.e. the indicator has to be
oriented to the action.
Integrated environment and health
surveillance systems have recently been
developed at international level.
The CDCs, the Californian Policy
Research Centre (CPRC), WHO-Europe,
and the Institut National de Santé Publique
du Québec have produced comprehensive
reports on the strategy to implement an
for environment and health monitoring
CNR Environment and Health Inter-departmental Project
The strengths and weaknesses of these
systems are summarized in Table 1 (10).
These three systems differ as for their
state of the art and completeness, but may
represent a good basis to initiate a program
of environmental and health surveillance
in Italy or in specific areas of the country.
3.1 USA
In the United States, the CDC ”Pew
Environmental Health Commission“
report first defined the purpose of a
surveillance system (17). This document
underlined the gaps in our knowledge of
environmental medicine and recommended
implementing a national environmental
health surveillance system.
In 2002, CDC in collaboration with
the Environmental Protection Agency
(US-EPA) and the National Aeronautics
and Space Administration and Member
Partners the development of this system
was started (18).
The goal of the system is to ”monitor
environmental hazards and disease trends,
advance research on possible linkages
between environmental hazards and
disease, develop, implement, and evaluate
regulatory and public health actions to
prevent or control environment-related
diseases“ (19).
In 2002, the State of California initiated
the first environmental health surveillance
system (20). The ”Strategies for Establishing
an Environmental Health Surveillance
System“ report led the early development
of this system.
It defined the objectives and usefulness of
developing and planning an environmental
health surveillance system, estimating its
costs, defining diseases, environmental
hazards and exposures to be monitored
and describing the related political, ethical
and legal issues (10).
In summary, the program of the CDC has
three objectives:
- to
infrastructure, such as the use of GIS
for mapping the use of pesticides and
the concentrations of pollutants in
urban areas;
- to improve data availability and use;
- to promote the translation of knowledge
into policy actions.
In this surveillance system, environmental
and health indicators (Environmental
Public Health Indicator, EPHI) are divided
into the following four categories (21):
- Hazard indicators: Conditions or
activities that identify the potential for
exposure to a contaminant or hazardous
- Exposure indicators: Biological markers
in tissue or fluid that identify the
presence of a substance or combination
of substances that could harm an
- Health effect indicators: diseases or
conditions that identify an adverse
effect from exposure to a known or
suspected environmental hazard.
- Intervention indicator: Programs or
official policies that minimize or prevent
an environmental hazard, exposure, or
health effect.
Indicators are also divided into three
a) pathways or sources (e.g, air, water);
b) agents (e.g. lead, pesticides);
c) events
environmental disasters).
Topics may also overlap due to the
complexity of environmental and public
health laws and programs. However, an
indicator is generally included under only
one topic, although it may be relevant to
Environmental Health Surveillance Systems
Table 1. CDC, EU and Quebec environmental health tracking systems: strengths and
Quebec (Institut national de santé publique du Québec 2006) 2004)
European Union (WHO Europe
Centers for Disease Prevention and Control (CDC
1. Partnership with federal, state
and local government agencies,
academic and community
groups, healthcare organizations
2. Strong stakeholder input
3. Pilot projects well
1. Includes upstream driving
2. Includes home, work and
ambient exposures
3. Includes population exposure
and health impact assessment
(air quality, noise)
4. Linked to health-based policy
action programs (NEHAPs)
5. Developing a children’s
environment and health
indicator set
1. Common surveillance with
occupational and infectious
diseases within Ministry of
Health and Social Services
2. Annual reporting
3. Research in environmental
health surveillance since 1997
with Geomatics for Informed
Decisions National Centre of
Excellence (GEOIDE NCE)
4. Strong public health
surveillance mandate in 2001
Public Health Law
5. Stable funding
6. Strong Quebec Public Health
Institute [Institut national de
santé publique du Québec
1. Varying levels of
state readiness
2. Early in
• First national
report, 2008
• Network launch
1. Diverse data
systems across EU
2. Gaps in survey
and biomonitoring
3. Still to define
outputs (printed
reports and Webbased data)
1. Not all indicators
2. Gaps in data for
some proposed
Air, ambient (outdoor)
Air, indoor
Lead (Pb)
Sentinel events
Sun and ultraviolet
Toxics and waste
Water, ambient
Water, drinking
Indicator Types
Health effect
160 indicators proposed in:
Air quality
Traffic accidents
Water and sanitation
Food safety
Chemical emergencies
Twenty-six of 41 indicators reported.
Environmental Indicators:
Recreational water quality (beaches)
Drinking water quality
Boil-water advisories
Waste water treatment
Air pollution
Environmental tobacco smoke exposure
Health-Based Indicators:
Carbon monoxide and other poisonings
notification rates
Allergic rhinitis prevalence
Cancers of interest for environmental health
Hospitalization/mortality rates for diagnoses
linked to environmental hazards
Proposed Indicators:
Indoor air
Climate change (mortality for heat waves,
morbidity and mortality linked to extreme
weather events)
CNR Environment and Health Inter-departmental Project
3.2 Canada
In the state of Quebec, the Ministry of
Health and Social Services has established
environmental hazards, occupational
health and infectious diseases. The
system is based on 26 of the 41 indicators
proposed by a panel of experts (17 refer
to environmental data, 9 to health data)
(Table 1).
3.3 European Union
The ECOEHIS project (European
Community Health and Environment
Information System), conducted under
the leadership of the WHO-European
Centre for Environment and Health (22),
has developed environmental health
(EH) indicators as part of the European
Community Health Indicators (ECHI),
which would serve as tools to aid in the
- To measure the health impact of
selected environmental risk factors,
their determinants and trends therein,
throughout the Community;
- To facilitate planning, monitoring and
evaluation of Community programs
and actions;
- To provide Member States and
information to make comparisons and
evaluate their policies (22).
The core set of environmental health
indicators has been developed within the
DPSEEA framework and focuses on the
population’s exposure to environmental
hazards, their health effects, and policy
actions to prevent illnesses, injuries and
Based on feasibility and usefulness testing
and after approval by the EU Member
States, the indicators were to be delivered
according to the evidence, data and
methodological limitations, in one of three
- ready
implementation (these indicators are
recommended as ‘core’ European
Community Health Indicators)
- ready, but not feasible for immediate
implementation (these indicators are
recommended for WHO projects such
- desirable but requiring further
developmental work (these indicators
The Institute for Environmental Protection
and Research (ISPRA) acting as National
Focal Point (NFP) for Italy, has coordinated
the national feasibility studies of these
indicators. Indicators refer to the following
- Air;
- Housing and health;
- Noise and health;
- Traffic accidents;
- Water and sanitation;
- Chemical Emergencies ;
- Radiation.
Based on the pilot project conducted in Italy,
ISPRA-APAT has classified the indicators
according to availability, data quality and
feasibility of their implementation for
three environmental sources (air, water,
soil) and for each of the five categories of
the DPSEEA framework.
Some indicators are not calculated due to
the unavailability of data or the gaps in the
informative flows.
The selected indicators are considered
of national importance, both in terms of
comparability and of data quality. They
need to be implemented and adapted at
local scale for surveillance in areas with
different environmental risks.
Environmental Health Surveillance Systems
3.4 Italy
In 2001, as part of the Italian Association
of Epidemiology, the Environmental
Epidemiology Group (GEA) was established
(23) in order to coordinate, organize and
take over environmental epidemiology and
risk assessment activities throughout the
country (24).
The environmental protection agencies that
have joined the group are: ISPRA, Regional
Environmental Protection Agency (ARPA)
Marche, ARPA Piemonte, ARPA Emilia
Romagna, ARPA Tuscany, ARPA Veneto,
ARPA Campania, ARPA Friuli Venezia
Giulia, ARPA Umbria, ARPA Lombardia,
APPA Bolzano , ARPA Basilicata, ARTA
Abruzzo, ARPA Liguria, ARPA Puglia,
ARPA Sicily, ARPA Sardegna.
As part of the GEA, the following four
subgroups have been formed:
- Group 1. Definition of guidelines for
environmental epidemiology studies
in small areas, in order to collect
information and experiences reported
in the literature for epidemiology
studies in small areas.
- Group 2. Integration between essential
health care levels (LEA) and the
Essential Levels of Environmental
Protection (LETA), in order to send
proposals to the Ministry of Health
and the Environment on LEA/ LETA
related issues.
- Group 3. Realization of a reference
epidemiology studies, in order to
promote organizational proposals
to create a network of experts on
environment and health.
- Group 4. Environmental and health
indicators at the local level (IAS).
It is based on the formulation of a
proposal for the definition and testing
of environment and health indicators at
local level (25).
These groups have produced ideas and
documents contained in the acts of the
second national workshop Portonovo
(Ancona) (26).
In the province of Brindisi, an area at “high
risk of environmental crisis” has been
identified by the Italian Ministry of the
Environment (Law n. 305 of 1989), due to
the presence of numerous industrial sites.
They produce a remarkable environmental
impact and cause serious alterations
of every type to the equilibrium of the
environment, as well as adverse effects on
the health of the population.
In fact, in the province of Brindisi and
particularly in the southern area of the main
town, on the Adriatic sea, many sources
of air pollutants with high environmental
impact are located near the urban area.
Next to the petrochemical area (built in
1959), various industries have grown up
over the years: three fossil-fuel power
plants, among them one of the largest
in Europe (Federico II Enel); several
chemical, pharmaceutical, metallurgical
and manufacturing industries; an airport;
an harbour, mainly for passenger traffic to
In 2002, in seven municipalities including
Brindisi, the State Forestry Service has
discovered 15 illegal dumps (covering an
area of 127,278 m2).
The Federico II Enel plant has the highest
record of CO2 emissions in Italy, and in
the area designed as Reclamation Sites
of National Interest (RSNI) there is a
significant concentration of particulate
matter as underlined by emissions and
concentrations recorded by the Region of
Puglia (CORINE-AIR).
CNR Environment and Health Inter-departmental Project
Southwest of Brindisi there is the province
of Taranto, whose industrial area includes
steel factories, a refinery and a cement
factory and proximity to this border
could be a further source of exposure to
environmental pollutants.
The town of Brindisi can be selected
as a site to test the surveillance system.
This requires epidemiological and
environmental characterization
4.1 Epidemiological characterization
Between 1990 and 1994, the World
Health Organization has conducted
an epidemiological study in four
municipalities (Carovigno, Torchiarolo,
S. Pietro V. and Brindisi) located in the
RSNI of Brindisi. Significant excesses of
mortality from all causes, all cancers, lung
cancers, respiratory and ischemic diseases
were observed both for males and females.
In particular, an elevated value of mortality
from melanoma was reported (27).
In Brindisi, in the male population,
excesses of mortality from all causes
and from all cancers were detected,
while in the female population excesses
were found for digestive system and for
psychiatric causes. Different mortality
patterns by gender are likely to be caused
by professional exposure.
A recent descriptive geographical study
of the province of Brindisi estimates the
mortality among residents in the twenty
municipalities of the province aggregated
in four geographic areas: the one at “high
risk” including the main town, and the
areas located north, west and south of the
Brindisi RSNI area (28).
The analysis was run by gender, specific
causes of death, and by two 10-yearperiods between 1981 and 2001. Results for
RSNI area confirmed the previous WHO
analysis, while ither excesses for specific
causes were observed in the remaining
In the province of Brindisi, excess mortality
due to cardiovascular disease and cancers
is higher than regional levels.
The analysis restructed to working age
groups (34-64 years), showed higher
rates of mortality than those reported for
cardiovascular mortality, among men as
well as women; excess mortality for cancer
of the prostate and for trauma was higher
in men, wheras women show a higher
mortality rate for the cancer of the central
nervous system. In addition, for Brindisi
Municipality, excess mortality for pleural
mesothelioma was reported also among
Table 2 shows the results of death incidence
analyses from all cancers and from specific
causes during the period 1999-2001, also
compared with the data of the whole
province. (the data source is the Jonico-
Table 2. Standardized rates of cancer incidence (x 100,000 inhabitants) and IC 95%.
Risk area of Brindisi
Province of Brindisi
CI 95%
CI 95%
CI 95%
All cancer sites
Urinary Bladder
Soft Tissue
Source:RTJS 1999-2001
Environmental Health Surveillance Systems
Table 3: emissions in atmosphere in Puglia
and in each of the five provinces – 2006.
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Source: INES registry
Salentino Tumor Register- RTJS).
In 2004 a case-control study was published
to investigate mortality from cancer in
the areas near Brindisi petrochemical
In period 1996-1997, a moderate excess of
mortality, from lung and bladder cancer
and lymphohematopoietic system was
observed in the population residing in an
area within 2 km from the centroide of
the petrochemical site, compared to the
population residing outside 5 km (29).
A case-crossover study has recently been
conducted to investigate the association
between daily mortality and hospital
admissions data, on the one side, and
the daily concentration of atmospheric
(PM10 and NO2) pollutants, on the other.
The study population included residents
in Brindisi city who died or hospitalized
for several diseases during the period
2003-2006 (30).
This study found strong and consistent
associations between outdoor air pollution
(coming from both industrial emissions
and urban traffic) and short-term increases
in both mortality and morbidity.
In particular, PM10 was associated with
mortality from all natural causes. The risk
was more pronounced for cardiovascular
hospitalization for cerebrovascular diseases
was statistically significant for PM10 among
females and elderly over 75 years old.
In specific population groups, increased
mortality and hospital admissions have
been associated with NO2 (31).
4.2 Environmental characterization
AIR characterization
The Italian Pollutant Emissions Register
(INES) registry can be used to gather
information about emission in water
and air coming from the facilities under
CNR Environment and Health Inter-departmental Project
Table 4: emissions in atmosphere in Brindisi by industrial complex – 2006
Meysure Unite
Kg / year
Mg / y
ENIPOWER S.P.A. - Brindisi
Mg / y
ENIPOWER S.P.A. - Brindisi
Mg / y
POWER PLANT Federico II (BR South)
Mg / y
Power plant Brindisi
Mg / y
Mg / y
Kg / y
Mg / y
ENIPOWER S.P.A. - Brindisi
Mg / y
ENIPOWER S.P.A. - Brindisi
Mg / y
POWER PLANT Federico II (BR South)
Mg / y
Power Plant Brindisi
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
Mg / y
C 6H 6
POWER PLANT Federico II/Brindisi
POWER PLANT Federico II/Brindisi
Source: INES registry
The Integrated Pollution Prevention and
Control (IPPC) Directive (2008/1/CE).
The emission concentrations are available
on the site:
In 2006, the Apulia region had the highest
emissions of all the pollutants considered,
against national data. The emissions could
be attributed mostly to the provinces of
Taranto and Brindisi (table 3).
CO2, NOx and SOx emissions - typical of
energy production - and benzene (C6H6)
emissions from the chemical sector mostly
originate from the province of Brindisi
(Tab. 3-4) .
WATER characterization
Table 5 shows the list of pollutants with
threshold values for each specific issue and
polluting industrial complex.
Environmental Health Surveillance Systems
Table 5: emissions in water in Brindisi from industry – 2006
Kg / year
Thresold value
Kg / year
Zn and compounds
ENIPOWER S.P.A. - Brindisi/Brindisi
As and compounds
POWER PLANT Federico II/Brindisi
Cd and compounds
POWER PLANT Federico II/Brindisi
Cu and compounds
POWER PLANT Federico II/Brindisi
Hg and compounds
POWER PLANT Federico II/Brindisi
Ni and compounds
POWER PLANT Federico II/Brindisi
Pb and compounds
POWER PLANT Federico II/Brindisi
POWER PLANT Federico II/Brindisi
Source: INES registry
SOIL characterization
In April 2008, ARPA-Puglia has published
a study on pollutants’ concentration in soil
near the industrial area of Brindisi.(31)
Soil samples were collected at five depth
strata (0-1, 1-2, 2-3, 3-4, 4-5) in 23 samplig
areas near the Federico II ENEL power
Concentrations of arsenic, beryl, tin,
cobalt, Chlordane, Vanadium, DDE, DDD
and DDT were measured.
The results show excesses of arsenic,
beryl, cobalt, tin, DDD in the sampling
points between 0 and 1 meter; excess of
beryllium and DDE in the top soil (layer
between 0-15 cm).
The arsenic concentrations in the five
sampling points (0 and 1 m) is between 24
mg/m3 e 53 mg/m3 (the threshold value in
residential area is 20 mg/m3). The arsenic
concentrations in 11 samples range from
a minimum of 2.2 mg/m3 to a maximum
of 3.8 mg/m3. In a top soil point the
concentration is 2.1 mg/m3 compared to
a limit value of 2 mg/m3 for residential
Cobalt values are always higher than
the threshold value with concentrations
ranging from 30 to 39 mg/m3 (the threshold
value in residential areas is 20 mg/m3).
Tin concentrations in four sampling points
(0 and 1 m) range from 1.2 mg/m3 to 5 mg/
m3 (compared to a threshold value of 1 in
residential area).
DDD values in the only one sample point
(0 and 1 m) are between 1.2 mg/m3 and
5 mg/m3 (the threshold is 0.1 mg/m3).
The concentration (0.63 mg/m3) exceeded
the threshold (0.1 mg/m3) in one on the two
samples (0-1 meter).
CNR Environment and Health Inter-departmental Project
A systematic proposal for sentinel events
regarding the environment and health
(ESAS) was drawn up by a consensus
conference organized by the Agency for
Toxic Substances and Disease Registry
(ATSDR) in early 1990s (32).
There are two types of events:
Type 1) Acute conditions as sentinel
indicators of environmental pollution, as
defined by Rutstein (33):
– Intoxication by pesticides, metals or
other substances present in refuse sites,
such as lead and carbon monoxide,
with particular attention to the most
vulnerable groups, such as children;
– Some tumors, such as pleural
cancer and hepatic angiosarcoma,
which although characterized by long
periods of induction-latency have
been associated with a high degree of
specificity for exposure to chemical or
physical agents;
– Precocious puberty as an indicator of
exposure to endocrine disruptors, such
as many pesticides, industrial products,
and food additives;
– Hemoglobinemia as a classic indicator
of intoxication;
– Neuropathy from exposure to toxic
chemical agents (such as methylmercury
causing Minamata disease).
Type II) Unusual models of disease
incidence or conditions identified by means
of surveillance:
– Bladder cancer, especially in nonsmokers and in the absence of
occupational hazards;
– Lung cancer in non-smokers;
– Liver cancer in non-drinkers;
– Rare tumors having a proven association
with environmental exposure, such
leukemia, acute leukemia in children
and granulocytic leukemia in adults;
Asthma in children, especially shortness
of breath in children in absence of
allergies and passive smoke exposure;
New environmentally-correlated rare
diseases such as eosinophilia-myalgia
syndrome, toxic oil syndrome, and
Kawasaki disease;
Measures of biological markers, such
as the concentration in biological fluids
of persistent organic pollutants (POPs);
DNA or hemoglobin adducts.
In environmental health surveillance,
especially in the case of small areas,
unusual events, low exposures, there is
a high statistical probability that many
warnings will be revealed by chance (due to
the effect of multiple testing). On the other
side, in case of alarm signals coming from
outside the surveillance system there is a
typical risk of an a posteriori selection.
No signals should ever be considered
conclusive, since investigation should
always be focused on statisticalprobabilistic confirmation and search for
the cause. No sign should be neglected
since it must be considered anomalous
until proven otherwise.
The adoption of a health surveillance
protocol for a polluted site is a suitable
tool to provide an answer to the legitimate
concerns of citizens regarding their
environment and the consequent health
impacts. The other crucial aspect of such
a surveillance system is the ability to
transfer reliable information and suitable
recommendation to decision-makers
in order to allow them to carry out and
evaluate political choices and programs on
the basis of scientific evidences.
Environmental Health Surveillance Systems
The environmental and epidemiological
characterization of Brindisi represents
the first step to set up environment-health
indicators, which represent the core of a
site-specific surveillance system.
With the contribution of WG5 participants: M.
Amadori (CNR-IGG), F. Bianchi (CNR-IFC),
(CNR-IFC), M.Cervino (CNR-ISAC), F.Cibella
(IBIM), P.Comba (ISS), L.Cori (CNR-IFC),
G. Latini (Local health Authority Brindisi),
P.Lauriola (ARPA-Emilia Romagna), A.
Mantovani (ISS), G.Petruzzelli (CNR-ISE), M.
Portaluri (Local health Authority Brindisi),
M.A.Vigotti (CNR-IFC).
Keywords: surveillance system, Brindisi,
environmental risk, health and environment
Thacker SB, Stroup DF, Rothenberg RB.
Public health surveillance for chronic
conditions: A scientific basis for decisions.
Statistics in Medicine 1995;14(5-7):629641.
Langmiur AD. The surveillance of
communicable disease of national
importance. N Engl J Med 1963;268:182192.
Thacker SB, Berkelman RL. Public
health surveillance in the united states.
Epidemiol Rev 1988;10(1):164-190.
Environmental epidemiology: A textbook
on study methods and public health
application. WHO 1999;WHO/SDE/
Thacker S, Stroup D, Parrish R, Anderson
HA. Surveillance in environmental public
health: issues, systems, and sources. Am
J Public Health 1996;86(5):633-638.
Organization for economic development
Environmental Indicators. 2007.
Corvalan CF, Kjellstrom T, Smith KR.
Health, Environment and Sustainable
and Indicators to Promote Action.
Epidemiology 1999;10(5):656–60.
Barcellos C, Quiterio LAD. Environmental
surveillance in health in brazil’s unified
health system. Revista De Saude Publica
Kyle AM, Balmes JR, Buffler PA, Lee
LP. Integrating Research, Surveillance,
and Practice in Environmental Public
Health Tracking. Environ Health Perspect
Abelsohn A, Frank J, Eyles J.
Environmental Public Health Tracking/
Surveillance in Canada: A Commentary.
Healthcare Policy / Politiques de Santé
Bianchi F. Prospettive di sorveglianza
ambiente e salute. Rapporti ISTISAN
European Environment Agency. EEA core
set of indicators - Guide. EEA Technical
report 2005;1.
Salute e ambiente – Quaderno di
Epidemiologia Ambientale. Osservatorio
Regionale Epidemiologico e per le
Politiche Sociali della Val d’Aosta. 2006.
Centers for Disease Prevention and
Control (CDC). Using indicators
for Environmental Public Health
Surveillance. Environmental Public
Health Indicators Project 2006.
Wills JT, Briggs DJ. Developing indicators
for environment and health. World Health
Stat Q. 1995;48(2):155-63.
Wcislo E, Dutkiewicz T, Konczalik
environmental hazards and health
effects in the industrial cities of upper
Silesia, Poland. Environ Health Perspect
Pew Environmental Health Commission.
America’s Environmental Health Gap:
Why the Country Needs a Nationwide
Health Tracking Network. 2000.
McGeehin MA, Qualters JR, Niskar
AS. National Environmental Public
Health Tracking Program: Bridging the
Information Gap. Environ Health Perspect
CNR Environment and Health Inter-departmental Project
19. Centers for Disease Prevention and
Control (CDC). National Environmental
Public Health Tracking Program
“Background”. 2006.
20. California Policy Research Centre.
Environmental Health Surveillance
System in California. A Report of the
SB702 Expert Working Group. 2004.
21. Centers for Disease Control and
Prevention. Environmental Public Health
Indicators. 2006:40 pp.
22. World Health Organization-European
Centre for environment and health.
Development of Environment and Health
Indicators for European Union Countries
- ECOEHIS. Final report 2004:655 pp.
23. Beccastrini S. Gli avvenimenti che hanno
preceduto la nascita del GEA e la stesura
del documento «Ambiente e salute»,
frutto del secondo convegno di Portonovo.
Epidemiol Prev 2005;29(3-4):139-140.
24. Documento conclusivo del Secondo
Seminario Nazionale “Integrazione
Ambiente e Salute”, Portonovo di Ancona
10 giugno 2005. Epidemiol Prev 2006
25. Aggiornamenti sulle attività del Gruppo
di lavoro GEA-AIE. Epidemiol Prev 2006
26. Mariottini M, Lauriola P. Secondo
ambiente e salute”. Portonovo di Ancona
Esperienze, proposte e dibattito per uno
sviluppo collaborativo della rete integrata
Servizio sanitario nazionale-Sistema
delle agenzie ambientali. Epidemiol Prev.
2007 Jan-Feb;(1 Suppl 2):1-78. Italian. No
abstract available. 2007;31(1 Suppl 2):1-78.
27. Martuzzi M, Mitis F, Biggeri A, Terracini
B, Bertollini R. Environment and health
status of the population in areas with
high risk of environmental crisis in Italy.
Epidemiol Prev 2002;26(6 Suppl):1-53.
28. 28 Gianicolo E A L, Serinelli M, Vigotti
M A; Portaluri M Mortalità nei comuni
della Provincia di Brindisi, 1981-2001.
Epidemiol Prev 2008;32(1):49-57.
29. Belli S, Benedetti M, Comba P,
Lagravinese D, Martucci V, Martuzzi
M, et al. Case-control study on cancer
risk associated to residence in the
neighbourhood of a petrochemical plant.
Eur J Epidemiol 2004;19(1):49-54.
30. Serinelli M, Gianicolo E A L, Cervino
M, Mangia C, Portaluri M, Vigotti
M A. Acute effects of air pollution in
Brindisi (Italy): a case-crossover analysis.
Epidemiol Prev. In press.
31. Valutazione dei rischi – Attività
agrotecniche area industriale di Brindisi,
fonte .
32. Shy C, Greenberg R, Winn D. Sentinel
Health Events of Environmental
Contamination: A Consensus Statement.
Environ Health Perpect 1994; 102 (3):
33. Rutstein DD, Mullan RJ, Frazier TM,
Halperin WE, Melius JM, Sesito JP.
Sentinel health events (occupational): a
basis for physician recognition and public
health surveillance. Am J Public Health
1983; 73 (9): 1054-1062.
Monitoring Contaminants in Food Chain
and their Impact on Human Health
A. Mupoa, F. Boscainoa, G. Cavazzinib, A. Giarettab, V.
Longoc, P. Russoa, A. Siania, R. Sicilianoa, I. Tedescoa, E.
Tostid, G.L. Russoa
a. CNR, Institute of Food Sciences (ISA), Avellino, Italy
b. CNR, Institute of Geoscience and Georesources (IGG), Padova, Italy
c. CNR, Institute of Biology and Agricultural Biotechnology (IBBA), Pisa, Italy
d. Zoological Station Anton Dohrn, Naples, Italy
[email protected]
In recent years a great attention has been focused in Europe on the importance of food safety and the relation
between diet and health. Moreover, worldwide changes in population lifestyle, together with modifications
in food processing, production and distribution contributed to the eating habits of Western populations and to
their reaction to recent public health emergencies. As an example, the real or alleged dioxin contamination that
affected several industrialized countries has increased the interest of Authorities, producers and consumers
on topics such as food safety and risks for human health deriving from contaminated food. The annual report
of the European Commission Rapid Alert System for Food and Feed (RASFF) summarizes notifications on
food contaminations occurred in different countries. Data analyses provided a useful tool to develop future
efficient programs for food control. In this context, a working group of the PIAS project studied how specific
classes of environmental contaminants (e.g, pesticides, metals, dioxins) may affect human health through the
food chain. Their results are presented in this Chapter. A special section has been dedicated to highlight issues
of major interest in this field, such as the determination of heavy metals and dioxins in food matrixes and
biological samples; experimental models to assess the harmful effect of contaminants on human reproduction;
the role of cytochrome P450 in xenobiotics metabolism. The last section of this Chapter proposes a research
programme aimed at integrating aspects already faced in current literature as independent issues, but rarely
considered in a holistic approach. The competences needed to pursue this goal are covered by the Italian
National Research Council or by the involvement of other Italian or international institutions. The final
proposal targets the youth and intends to determine the cause-effect relationship between the presence of
contaminants in the diet, their accumulation in humans and the risk of chronic diseases. Key issues, such
as bioavailability and adaptive response (hormesis), will be explored using suitable experimental models to
suggest a functional link, at molecular level, between the onset of specific diseases and the concentrations of
contaminants measured in food.
1.1 Food safety: focusing on chemical
The introduction of genetically modified
food and food incidents in Europe raised
the public interest in food safety. An
integrated approach to face this problem
requires a strong cooperation by the food
industry, food distributors, the scientific
community, governments, managers and
local administrators in order to build
consumers’ trust and confidence. The food
safety certification is achieved assessing
the potentially health adverse effects of
food contamination. Three main food
contamination groups can be identified: i.
physical; ii. microbiological; iii. chemical.
Physical contaminations are due to the
presence of extraneous bodies in food
(plastic, woods, glass and others) as the
results, for example, of food packaging
and/or transformation and/or storage.
The substances present in those materials
are not for human consumption, but
CNR Environment and Health Inter-departmental Project
when in contact with food they migrate
into it and risk of being ingested (for
example, the perfluorinated chemicals
used in greaseproof packaging for fast
foods). Microbiological contamination
refers to the presence of one or more
natural biological agents, such as various
bacteria, yeasts, mould, fungi, protozoa
or their toxins and by-products, which
can adulterate food properties and safety.
Microbiological agents are responsible
for “food diseases” such as food borne
infections and intoxications (Botulinum,
Listeria, Hepatitis A) and epidemic
episodes (e.g, Salmonella enteritidis).
Chemical contaminants or xenobiotics
can originate from many different sources
and include heavy metals, pesticides,
additives, dioxins and PCBs. Nowadays,
chemical contaminants are a major concern
for food safety because of the increased
role of man-made chemicals due to our
modern lifestyles. In fact, despite the fact
that the large use of chemicals improved
the quality of our lives, many of them
have been reported to produce an adverse
impact on human populations, animals and
plants continuously exposed to a cocktail
of potentially hazardous chemicals (2-4).
However, in humans and animals, diet is
predominant route of many dangerous
chemicals. Food is a crucial link in the
chain of events starting from chemical
manufacturing and ending with their
presence in human blood, tissues and
organs. (5). The worldwide observation
of such contaminants in food shows the
global scale of chemical contamination.
To assess the impact of such substances
on food safety, the following question
must be answered: Can the quantity and
bioavailabilty of an unwanted chemical
in food provide a real risk to human
health? In the past, just the presence of
a hazardous chemicals, whatever their
concentration or weight, was considered as
unsafe and adverse to health. The presence
of a chemical depends on the sensitivity of
the instruments used to assess it. Analysis
with an increased sensitivity and different
techniques showed that some chemicals,
previously stated as not present, were
instead only undetected.
This implies the detection itself is not
necessarily representing a risk: a new
approach is needed to provide a riskbased evaluation of the potential exposure,
hazard, and toxicity of chemicals detected
at a low-level (6).
For this reason, a threshold is needed for
non toxic chemical compounds. In past
decades, scientists developed different
models to address this issue, concluding
that the potential human health risk posed
by a chemical substance depends on its
inherent toxicity and exposure, including
route, dose and duration. In the case of
substances found in food, at least two
elements must be considered. The first is
their concentration level in various foods,
assessed through chemical analysis. The
second element is the consumed quantity
of contaminated food. Bioavailability
must be also considered as the capability
of a dietary chemical to be absorbed and
metabolized. Bioavailability is commonly
assessed by measuring the amount of
the ingested chemical that gets into the
systemic circulation, since in most cases
the specific targets and the time required to
determine an effect on health are unknown.
Thus, although bioavailability is critical in
assessing the potential benefits or risks of
a compound, it can only be studied on a
comparative basis (see section 5).
It is clear that great effort is devoted to
improve risk assessment and to develop
common methods to be used to guarantee
food quality and to protect consumers’
Monitoring Contaminants in Food Chain and their Impact on Human Health
health. Such assessments are often based
on very limited scientific information and
a complete and exhaustive data set to be
used to provide definitive conclusions on
chemical concerns in food does not exist
at the moment. In this context, results from
innovative research programmes are the
main sources of information to establish
a common food policy and widen the
existing data set.
1.2 Contamination chain: the food link
substances present in the environment
where food is grown, harvested,
transported, stored, packaged, processed,
and consumed. This “food chain route”
of contamination implies the presence of
different food contamination levels (Fig.
1), an important element to be evaluated
in food safety. For example, after they
are released into the environment (soil,
air, water) chemical contaminants can
enter plants and animals at the bottom of
the food chain, to be then consumed by
animals, going up in the same chain. The
chemicals contained in animals and plants
can enter human bodies through the diet.
This concept is even more important for
persistent chemicals (biomagnification) and
accumulated chemicals (bioaccumulation)
(e.g, pesticides, dioxins or heavy metals).
These compounds are called Persistent
Organic Pollutants (POPs) and include all
those substances not rapidly degraded that
keep their harmful capability towards both
the environment and human health.
These substances have the following
properties: i. resistance to degradation; ii.
Long time persistence into the environment;
iii. toxicity for humans, animals and plants;
iv. accumulation in living organisms.
Bioaccumulation implies that the
compound is lipid soluble and, in the
absence of an adequate metabolic pathway
able to eliminate it from the organism,
tends to accumulate in the trophic chain.
For example, polychlorinated biphenyls
(PCB) are very stable organic compounds;
they are highly persistent and present in
air, soil and water; they are lipid soluble
and bioaccumulate in animal fat, in meat
and in the liver, and are transferred into
milk and eggs. More than 90% of human
exposure to PCBs derives from food of
animal origin (7).
Figure 1. “The Food Chain Route”
CNR Environment and Health Inter-departmental Project
Biomagnification is the process by which
a compound increases its concentration
along the food chain. Heavy metals can be
bioconcentrated along the trophic chain.
These substances can be involuntarily
ingested with food and drinks and,
once absorbed, they are distributed in
tissues and organs, persisting for years
or decades in some storage sites such
as liver, bones and kidneys. Inorganic
mercury, for example, can be converted
by water micro organisms into the organic
methyl mercury compound, which is then
biomagnificated in higher links of the food
chain. Fish, especially tuna or swordfish,
can concentrate methyl mercury at high
levels (8).
1.3 Chemical contamination and human
A major concern about food contaminants
is their possible adverse effects on human
health. Reports on human illnesses caused
by food toxic contaminants began several
centuries B.C, and since then numerous
episodes of food diseases have been
continuously reported (9).
In recent years, many of these chemicals
present in food have been detected in
the blood, tissues and organs of children
and adults. POPs are responsible for
nervous systems syndromes, disruption
of infant brain development, immunepathologies,
abnormalities, cardiovascular diseases,
cancers, diabetes and obesity, and some
of them can act as endocrine disrupting
agents. The effect on human health can be
classified according to: i. acute exposure
(early effects); ii. chronic exposure (long
term); iii. foetal and infants exposure.
Acute exposure implies the exposure to
a massive dose of the contaminant and its
negative effects on health are immediate
(e.g, milk contaminated with melamine).
Chronic exposure implies a long term
contact with the contaminants before the
disease is manifested (e.g, heavy metals).
An issue that has recently become a
priority is related to the negative effects on
normal development of foetus and infants
exposed to contaminants through the food
chain (e.g, POPs). According to the U.S.
Environmental Protection Agency (EPA)
Toxic Substances Control Act list (10), there
are more than 75’000 known chemicals in
the environment, many of which may enter
the food chain. Due to the complexity
and the huge amount of information, this
section will be focused on some specific
compounds (including some heavy metals,
dioxins and pesticides) to evaluate their
impact on human health.
1.3.1 Heavy metals: lead, mercury, arsenic
and cadmium
Metals are natural elements that have been
used in human industry and products since
millennia due to their chemical and physical
properties. Metals can be easily dispersed in
the environment, in soil, water and air and can
be very toxic even at relatively low levels of
exposure; moreover they can accumulate in
specific tissues of the human body.
The U.S. Agency for Toxic Substances and
Disease Registry (ATSDR) produced a
complete list of the hazards present in toxic
waste sites according to their prevalence
and the severity of their toxicity: “heavy
metals” (lead, mercury, arsenic, and
cadmium) are at the top of this list (8).
The lead found in food is present as salt or
oxide and only a small fraction is adsorbed
by humans (up to 10%). Lead toxicity can
be acute or chronic. Acute intoxications
are unusual but are responsible for gastrointestinal,
(anemia) and nervous system (convulsions)
symptoms. Chronic exposure is generally
manifested with anemia, which depends
on the direct toxic effect of lead on red
Monitoring Contaminants in Food Chain and their Impact on Human Health
blood cells and bone marrow. It can cause
toxic effects also on the nervous system
(hyperkinesia, parlysis) and renal failure.
Everyone can be affected from lead
toxicity, but infants and fetuses are more
vulnerable to lead exposure and can suffer
serious damage to the development of their
nervous system and learning disabilities.
Mercury is a chemical element widely used
in scientific equipment (e.g. thermometers,
barometers). However, mercurous and
mercuric mercury can form inorganic
and organic compounds with other
chemicals and can be readily absorbed
through ingestion. At high levels, mercury
poisoning is responsible for injuries to the
lungs and the neurologic system. At lower
levels, mercury poisoning is responsible for
erethism (tremor of the hands, excitability,
memory loss, insomnia, timidity, and
sometimes delirium).
Exposure to low doses of mercury is of
great concern for its effects on the nervous
system development in fetuses and infants.
In 1955 after the disaster in Minamata Bay,
Japan, local doctors and medical officials
noticed for a long time an abnormally
high frequency of cerebral palsy and
other child disorders in children born in
the area (congenital Minamata disease).
Moreover, studies in the Faroe Islands
have demonstrated that, even at much
lower levels, mercury exposure of pregnant
women, through dietary intake of fish and
whale meat, is associated with decrements
in motor function, language, memory, and
neural transmission in their offspring (1112). Organic mercury, the form of mercury
bioconcentrated in fish and whale meat,
readily crosses the placenta and appears in
breast milk.
Exposure to arsenic originates from
anthropic industrial activities and the use
of products such as wood preservatives,
pesticides, herbicides, fungicides, and
paints. In some areas of the world, arsenic
is also a natural contaminant of water.
Moreover, arsenic can accumulate in
seafood. Once absorbed into the body,
arsenic undergoes some accumulation
in soft tissue organs such as the liver,
spleen, kidneys, and lungs, but the major
long-term storage site for arsenic is
keratin-rich tissues, such as skin, hair
and nails. Acute arsenic poisoning is
infamous for its lethality, since arsenic
destroys the integrity of blood vessels
and gastrointestinal tissue and its effect
on the heart and brain are huge. Chronic
exposure to lower levels of arsenic results
in somewhat unusual patterns of skin
hyperpigmentation, peripheral nerve
damage, diabetes, and blood vessel damage
(13). Chronic arsenic exposure also causes
a high risk of developing a number of
cancers, in particular skin, liver lung,
bladder, kidney and colon cancers.
Cadmium pollution (e.g, the emissions
from cadmium smelters or industrial
emissions and the introduction of
cadmium into sewage sludge, fertilizers,
and groundwater) can result in significant
human exposure through the ingestion
of contaminated foodstuff, especially
grains, cereals, and leafy vegetables. Once
ingested, cadmium is adsorbed in the
gastro-intestinal tract and accumulates
in liver and kidneys. Acute high-dose
exposures can cause severe respiratory
irritation. Lower levels of exposure are
worrisome mainly for their kidney toxicity.
Even without causing kidney failure,
cadmium effect on kidneys can have
metabolic and pathologic consequences. In
particular, the loss of calcium caused by the
effect of cadmium on the kidneys can be
severe enough to lead to bone weakening
(osteoporosis, osteomalacia) (8).
CNR Environment and Health Inter-departmental Project
1.3.2 Pesticides
Pesticides are a class of chemical
compounds used in agriculture to fight
parasites and other organisms dangerous
for plants, animals and humans. They are
divided into different classes of molecules
according to their properties as inorganic,
natural organic and synthetic organics.
Synthetic organic pesticides are the most
used and include DDT, DDE, aldrin, dieldrin
and others. DDT (diclorofenicloroetan)
was synthesized in 1940 and used in
agriculture against many insects. Due to its
high toxicity and high persistence, the use
of DDT is now banned in most countries.
The major concern on the use of pesticides
is related to their carcinogenic effect,
their activity as endocrine disruptors and
their neurotoxic effects. Epidemiological
studies on workers (agriculture) in
contact with these substances showed an
increased risk for their health safety (14).
Scientific evidence showed that many
pesticides used today have neurotoxic
activity. For example, commonly used
organophosphorous pesticides can inhibit
the acetylcholinesterase (AChE) function,
the enzyme that degrades the acetylcholine
neurotransmitter in the central and
peripheral nervous system. Acetylcholine
can then accumulate in the nervous system
producing an unwanted nervous response
that can be responsible for paralysis, muscle
debility, convulsions and, sometimes,
death (15). Moreover, the use of some
fungicides (mancozeb, maneb), that are
rapidly metabolized in the organism and
in the environment, can generate a highly
toxic product, etilentiourea (ETU), that
interferes with the thyroid functions and
can induce malformations in fetuses when
exposed to high doses (16).
In Europe, the legislation on the use of
pesticides is complex and articulated.
Very recently, the European Commission
officially adopted and published a new
regulation setting in motion major changes
in how plant protection products are placed
on the market and how they are used in
practice. Essentially, the new regulation
will forbid some ‘active substances’ in
pesticides. In particular, the European
Parliament says, the legislation seeks to
outlaw highly toxic chemicals, such as
those which cause cancer (17).
1.3.3 Polychlorinated biphenils (PCB)
Polychlorinated biphenyls (PCB) cover a
group of 209 different congeners, classified
according to their number and position
of their chlorine atom substituents. Most
important, PCB are highly persistent,
are globally circulated by atmospheric
transport and therefore are present in all
environmental media. Due to their lipid
solubility and the absence of adequate
metabolic pathways in the organisms,
PCB tend to bioaccumulate along the
trophic chains. As a consequence, PCB
are major components of POPs together
with polychloro-dibenzodioxins and
(7). In general, human exposure occurs
trough the diet, particularly through the
ingestion of meat, fish, milk and other
dairy products, while in industrial areas
showing dioxin emissions the inhaled
component has a greater importance.
There is a lot of concern on the negative
role of dioxins on human health. Dioxins
toxicity has been related to different types
of cancer, endocrine interference, deficit in
the immune response and developmental
defects in fetuses. However, studies on
children indicate that the exposure of
the general population to low levels of
polychlorinated PCDD/Fs does not result in
any clinical evidence of disease, although
accidental exposure to high levels either
before or after birth have led to a number of
Monitoring Contaminants in Food Chain and their Impact on Human Health
developmental defects (18): Experimental
data indicate that the endocrine and
reproductive effects of dioxins should be
among the most important effects in animal
and humans (19,20). Nowadays, the debate
is still ongoing on the real toxic effect of
dioxins after low-level exposure, an issue
that needs to be further investigated.
2.1 Food quality control
Quality is defined as any of the features
that make something of a degree of
excellence or superiority (21). The word
“quality” is differently used in food
science and technology referring to a
complex concept which includes, on one
side, characteristics related to nutritional,
microbiological and chemical properties
as evaluated from food experts (22) and,
on the other side, the sensory quality of a
food defined as the attributes of the food
which make it agreeable to the person who
eats it (the consumer). The latter involves
positive factors like color, flavor and
texture (23). Quality control is the sum of
all those controllable factors that ultimately
positively or negatively influence the
quality of the finished product, e.g, selection
of raw materials, processing, packaging,
storage and distribution methods. All
along the supply chain, food is exposed to
numerous hazards. To prevent or mitigate
most of them, the risk factors present at
each phase of the supply chain must be
known and an effective and comprehensive
quality system must be in place. The aim
of quality control is to achieve good and
consistent quality standards compatible
with the market for which the product is
designed. Food quality control implies the
control of different food processing steps
to prevent the adulteration of the final
Some of the most important steps that need
to be evaluated in food quality control are:
i. agricultural materials / ingredients; ii.
processing / engineering; iii. additives; iv.
packaging; v. finished product inspection
(24). Problems may arise in some of those
phases, having a negative impact on the
finished product: they are critical points
The first CP in food control concerns soil
quality. In fact, food can be contaminated
at a very early stage in the food chain and
the contamination propagates all the way
along. Soil quality can be evaluated in two
distinct ways: i. as an inherent characteristic
of a soil; ii. as the “health” condition of
the soil (25). The former includes some
parameters that reflect the potential of a
soil to perform a specific function, (i.e.:
plant growth and production, quality of the
plants and fruits, soil natural resources).
The latter includes agricultural practices
that can adulterate ground functions and
composition, such as manure, the use of
fertilizers and pesticides, but also manmade chemicals or other contamination
that usually arises from direct industrial
waste discharge into the soil, percolation of
contaminated water or wind contamination.
The most common chemicals involved
are PCBs, solvents, lead and other heavy
metals. Soil quality control is performed
by environmental scientists in compliance
with generic guidelines that include field
measurement, also using computer models,
to evaluate the minimal acceptable level
of a substance and eventually determine
the clean up options for the contaminated
In the “food processing” industry,
raw materials are the main source of
contamination. Stores and warehouses
often make a large use of a wide range of
CNR Environment and Health Inter-departmental Project
raw materials. Every product has one (or
several) dominant raw material on which
the quality of the finished product mainly
depends (26). Raw materials control is
another CP to ensure food quality and to
be performed it requires the use of specific
sampling. The formulation of the sampling
type and test applied must reflect in the
finished product and must be fast, simple
and suited to the purpose. These tests can
be chemical, physical, bacteriological or
organoleptic and are usually performed
in those specific laboratories that can
authorise the factory to use the raw
Finally, food packaging control is needed
in order to protect consumers from the
package to foodstuff migration of harmful
substances. Nowadays, packaging is an
essential element in food manufacturing
processes because it gives food more
safety and a longer shelf life. In Europe, the
Commission of European Communities
(CEC) controls and establishes directives
for the use of plastic packaging materials.
In general, these directives are based on
analytical test methods to establish the
limits of plastic-package migration into
food. These analytical procedures are
used: i. to identify the potential migrants
and their toxicity; ii. To identify the factors
responsible for migration: iii. To estimate
the intake of food contaminants; iv. To
determine the level of contaminants in the
packaging materials and in the food they
have been in contact with (27).
After the manufacturing process, food
quality can not be modified. Thus finished
products examination can only grant
acceptance to materials reaching the
desired standard or rejection to materials
failing to reach this standard.
Food quality control is a concept which
evolves as experience and knowledge in
the field grows. In the modern world, all
food processing undergoes quality control,
often based on discoveries derived from
basic research in the field. From these
observations, it can be argued that in
the future there will be a possibility to
generate a unique control model and, using
modern data processing methods, obtain a
continuous monitoring of the events during
all the phases of the food production flow.
2.2 Dietary exposure to contaminants:
total diet studies
From the information collected so far, it is
clear that there is a general concern on food
quality, contaminations, safety and effects
on human health. Moreover, there is the
need to put together all information coming
from different sources (government, food
scientists, local agencies and others) in
order to define the best approach to prevent
food diseases and to establish general rules
for a “better food”. A strong contribute in
this direction comes from both basic and
applied research. There are three key steps
that must be considered when defining a
scientific approach to food contamination
as proposed in Thacker’s model (Table 1)
Table 1: Thacker’s model
Pesticides, Heavy
metals, PCBs
Risk/source of
Risk evaluation
Effects on human
First, the contaminant must be identified;
then, the source through which the
contaminant could reach the consumers
must be identified; finally. the adverse
Monitoring Contaminants in Food Chain and their Impact on Human Health
effect (hazard) of the contaminant must
be determined in order to prevent and/
or protect exposed individuals. In this
context, the present paragraph will analyze
some examples coming from the scientific
literature regarding elegant approaches
applied in different countries to estimate
human exposure to food contaminants
trough the diet (Total Diet Study). In
particular, the case of dioxins will be
considered. This topic concept will be also
discussed in section 5 from a different
point of view.
Bilau and co-workers (29) have recently
carried out an important study on three
age groups of the Flemish population,
adolescents (14-15 years), mothers (18-44
years) and adults (50-65 years) to determine
the intake of dioxin-like compounds via
animal fats or other sources, namely dairy
products, added fats, fish and seafood.
The study was performed by assessing
the dietary intake of all the participants
to the study using a semi quantitative
food frequency questionnaire (30). The
questionnaire has been used to estimate
the daily consumption of fat-containing
food items for each participant and,
based on their dietary habits, the intake
of fat from the different sources (meat,
fish, dairy products) was determined.
Contaminant concentration (dioxin and
dioxin-like compounds) was measured
in food items coming from the Flemish
market, via the chemical-activated
luciferase gene expression (CALUX)
bioassay (31). To estimate the dietary intake
of dioxin-like compounds in the studied
population a simple approach distribution
was used, combining a point estimate
for contaminants concentration with the
distribution of individual consumption
data (32). The result of the studies shows
that a large part of the three study groups
exceeded the weekly “safe” intake of
dioxin-like compounds and also that this
intake decreased with age. Moreover, in
the Flemish population fish and seafood
resulted as the main source of dietary
intake of dioxin-contaminants.
In another study, the same approach was
used for Swedish children and adults
showing that children are a vulnerable
group with a daily over-intake of dioxins
from food commodities in particular from
fish. For this reason, the authors suggest
that it should be useful to perform agespecific dietary intake assessments to
protect highly exposed individuals (33).
Similar studies have also been conducted in
other European countries with comparing
results (see section 5).
However, a key aspect emerging from this
scientific work is that in most of the studies
only two of the three steps foreseen by the
Thacker’s model have been considered: the
identification of the contaminant or hazard
is fulfilled (e.g, dioxins in foodstuff)
and the diet is identified as the source of
contamination. It lacks the proof of concept
that this low-dose exposure deriving from
food and assessed by the Total Diet Study
is really responsible for effects on human
health (see below on section 5).
In many cases, there is a general
assumption that the mere presence of the
contaminant will affect or be harmful for
consumer safety, now or in the future. It is
clear that casual exposure to high doses of
contaminants generally represents a threat
for human health (e.g, dioxin exposure
in Seveso population). However, the low
exposure impact of some contaminants,
such as dioxins, is still debated. An
interesting example on this specific
point comes from the Food and Drug
Administration (FDA) website regarding
questions and answers about dioxins: Q:
“What levels of dietary dioxin exposure
cause adverse health effects in humans?”;
CNR Environment and Health Inter-departmental Project
A: “Known incidents of high dioxin levels
in humans have resulted from accidental
exposures that are not typical with dietary
exposures. Despite a large body of research
and data collection, there are numerous
questions and uncertainties regarding
scientific data on and analysis of dioxin
risk. These uncertainties are unlikely to
be resolved in the near future” (www.fda.
3.1 General overview on food agency: an
eye on Europe and Italy
“There are certain things only a government
can do. And one of those things is ensuring
that the food we eat is safe and does not
cause us harm.” (President of United States
of America, Barack Obama).
Food always had a strong influence on
daily life and production/consumption
of food is central to any modern society.
For this reason, at the heart of any foodrelated topic there is the need to consider
the citizen/consumer as the final “user”
of the total food/feed chain and the one
who needs to be protected from any risk
of disease. In Europe, the main agency
that controls risk assessment regarding
food and feed safety is the European Food
Safety Authority (EFSA): “EFSA aim is to
provide appropriate, consistent, accurate
and timely communications on food
safety issues to all stakeholders and the
public at large, based on the Authority’s
risk assessments and scientific expertise”
( The main mandate
of EFSA is related to risk assessment and
risk communication. Risk assessment is
a specialized field of applied science that
involves the analysis of scientific data and
studies in order to evaluate risks associated
with certain hazards. This implies scientific
data collection and analysis on a wide
variety of hazards (e.g, pesticides, PCBs,
microbiological agents and others) to gather
information on dangers posed from these
substances, to develop general methods to
assign a date risk for the consumer. One
of the key responsibilities of EFSA is to
communicate food and feed safety advice
to its principal clients, stakeholders and the
public in a timely, clear and helpful way,
in order to help bridge the gap between
science and the consumer.
It is clear that due to the complexity of
this issue, and, what is more, the different
sources of information ranging from
local to international agencies, a key step
concerning food safety is the possibility to
exchange information among controlling
agencies. Nowadays, the Rapid Alert
System for Food and Feed (RASFF) in
Europe represents a powerful tool to
exchange data about measures taken in
response to serious risks detected in food
or feed. There is a very simple principle
at the basis of the RASFF system:
“Whenever a member of the network has
any information relating to the existence
of a serious direct or indirect risk to
human health deriving from food or feed,
this information is immediately notified
to the Commission under the RASFF.
The Commission immediately transmits
this information to the members of the
network” (34) (Fig. 2) .
In Italy, the main actions concerning
food quality and control derive from
government agencies, local offices and/
or private companies. Most information
coming from the government is
released by the Ministero delle politiche
agricole alimentari e forestali (www., and the National
Institute of Health ( At local
level, control and communication roles
are mainly played by the Environment
Monitoring Contaminants in Food Chain and their Impact on Human Health
Protection Regional Agencies (ARPA), the
Local Health Authorities (ASL), the National
Agrifood Informative System (SIAN) and
others, often working in collaboration with
the local police. All these agencies develop
specific actions and projects to understand,
prevent, control and reduce the risk related
to food contamination.
Several institutes of the National Research
Council ( are involved in these
tasks from different points of view: i. to
develop new methods and strategies for
analysis; ii. To apply for national and
international research projects in the
field; iii. To establish collaborations and
consultation with official agencies devoted
to control activities. In this context, one of
the actions in the PIAS project has been to
better categorize data in the field of “food
chain contamination and effects on human
health” originated from the scientific work
performed by CNR research groups, an
issue that will be further described in the
following section.
3.2 CNR research activities: results from
PIAS questionnaire
The CNR is a public organisation
promoting, transferring, communicating
and enhancing scientific research in
different fields to improve the country’s
technological, economic and social
activities. The organization is divided into
eleven Departments, also in accordance
with the research work performed at CNR.
All the relevant projects developed at CNR
can be viewed surfing the CNR web sites
Due to the huge amount of information and
the many different scientific topics that are
part of the research activities performed in
the CNR, sometimes it could be very hard
to gain data on the field of interest. In this
view, one of the aims of the PIAS project was
to clarify, and eventually harmonize, the
activities of the different scientists in CNR
trough communication, data exchange and
eventually collaboration. In our survey,
out field of interest was the “monitoring
of environmental contaminants in the food
chain and their impact on human health.”
To acquire information on the activities
related to this topic at CNR, we elaborated
a questionnaire which was send to the
main Departments and Institutes involved
Table 2. Summary of the information
obtained from the PIAS questionnaire
Biochemistry, GCMS, HPLC-DAD,
analytical chemistry,
Gas chromatography,
molecular biology, cell
biology, bioinformatics,
biomarkers, massspectrometry
POPs, pesticides,
heavy metals, organic
compounds, toxins,
Pasta, bread, milk,
cereal, fruit, vegetables,
disease, inflammation,
fitness, cancer,
disease, toxinfections,
lung disease, genetic
disease, immune
CNR Environment and Health Inter-departmental Project
in our field of interest. Key points in the
schedule were represented by the group
composition, expertise, methodologies
applied, type of contaminant/food studied,
type of pathologies analyzed and main
projects developed by the research groups.
The data elaborated from the questionnaire
are summarized in Table 2.
From Table 2, we can assume that CNR
possesses specific expertise originated
by the different groups working on the
indicated contaminants, present in food
matrices reported in Table 2 by using
specific methodologies spanning from
cell biology to mass spectrometry. This
approach allows the performance of a risk
assessment related to specific diseases. It
is clear that the data reported in Table 2
probably represent only a fraction of the
real competence present at CNR. This
underestimation is probably due to an
incomplete feedback received from the
questionnaire we send. However, the
data collected show the existence of the
capabilities required to fulfill Thacker’s
model, which are already part of CNR
scientists’ cultural background. This is a
key point, because we can speculate that, in
a near future, it will be possible to develop
a global, collaborative project in the field
of food quality and safety related to the
presence of environmental contaminants
by merging the different competences
coming from the different CNR groups.
Heavy metals in food: traditional
and innovative detection methods
Metals are constituents of the human
body and some of them are fundamental
for body growth, metabolic reactions and
catalysis mechanisms. For this reason they
are considered as essential constituents
for life, and are distinguished in ‘major’
or ‘minor’ (trace), depending on their
concentration levels (35-39).
However, the role that some metals play
in the human body is not completely
understood. Some metals have been
recognized to be certainly dangerous for
human health. Cd, Hg, Pb, As, Ni, Al, Cu
may cause illness, aging and even genetic
defects, and mankind is today exposed to
the highest levels of these metals due to
their use in industry, to the unrestricted
burning of coal, natural gas and petroleum,
and to the incineration of waste materials
worldwide (40-48).
The term heavy metals is often used in
current literature to indicate the toxic
metals as a group, although heavy refers
to mass (it should be actually used only
for elements the atomic weight of which is
higher than 200 such as mercury, thallium,
lead and bismuth) and mass does not seem
directly related to toxicity (49,50).
Plants contamination occurs when heavy
metals are present in the soil where they
are grown, and animals that are fed with
these plants are also contaminated (51,52).
The quantities of the different elements
in soil generally vary from place to place,
and the amounts absorbed by plants and
retained in their tissues can also show
large variations (15,53-60). Therefore,
there can be considerable variations
in concentrations and also in isotopic
composition of metals even within the
same class of food, depending on its
geographical origins and other factors.
For these reasons, concentration and/
or isotopic ratios of metals can be used,
sometimes with success, as indicators of
food provenance (61-71).
Changes in concentrations and isotopic
composition of metals in food may be
not confined to primary, geographicallyrelated variations. They also may be due
to food manufacture, and in some cases,
Monitoring Contaminants in Food Chain and their Impact on Human Health
Figure 2. Schematic representation of the information flow of the RASFF
trace of metals can cause undesirable preceded by exchange chromatography)
changes in food during cooking and/or (109-112), with ICP-MS (113,114) and with
storage (72-75). All these factors may have GC-MS (115).
consequences on the consumers; therefore, In some of these techniques (AAS-F/GF,
many government authorities have specific ICP-AES/MS, ID-TIMS/ICP-MS), samples
rules for food manufacture.
are reduced to a perfectly homogeneous
In the last two decades, analytical solution before the instrumental analysis,
techniques and instrumentation to with the exception of liquids, such as
determine concentrations of metals in beverages (including water) which may
foods have been improved.(76-80). A only require dilution. In most foods,
limited number of methods are mainly organic matter must be removed by
used in this field and they include: oxidation because it would interfere with
Atomic Absorption Spectrophotometry the analytical process, either by the use of
(AAS) (81-86), Spectrofluorimetry, Gas oxidising acids in a wet digestion or by dry
Spectrometry ashing in the presence of air or pure oxygen.
(GC-MS) (87,88), Inductively Coupled Virtually, all organic matrices of food can
Plasma-Atomic Emission Spectrometry undergo these two different preparation
(ICP-AES) (80,83,89-91), Inductively procedures and the choice depends mainly
Coupled Plasma-Mass spectrometry (ICP- on the metal(s) to be determined. Dry
MS) (83,89,92-108), Isotope Dilution ashing technique at 550°C causes Hg, Sn
combined with Thermal Ionization Source and As loss by evaporation. Thus, for these
Mass Spectrometry (ID-TIMS) (usually elements sample digestion with HNO3
CNR Environment and Health Inter-departmental Project
and H2O2 in closed PTFE vials within a
microwave system is the typical preferred
AAS is a very versatile technique which
does not need a particular laboratory and
instrumental condition, and AAS-GF
may allow determinations at ultra-trace
concentration levels (<1 ppm). However, in
this technique, only one element at a time
can be determined, and, due to relatively
low operative temperatures, determination
of the refractory metals (e.g, REE, Sc and
Y) is substantially precluded (83).
Due to definitely higher temperature in
the torch, ICP-AES/MS is a powerful tool
to determine all the metals, including the
refractory ones, combining high sensibility
with considerable accuracy and fastness
(80,83,89-108). In particular, ICP-MS, by
rapid determination of isotope ratios of
elements, can be combined with isotope
dilution technique in multi-collector
increasing the precision and the accuracy
of the determinations (113,114,122,123).
At present, ICP-MS technique has
considerably extended its capabilities,
by combining with different separation
procedures, as chromatography and
electrophoresis, and by developing of
methods of sample introduction, such as
flow injection, thermal vaporisation and
laser ablation (92,95-100,106,124-126).
Unlike the AAS system, however, the ICP
systems require quite rigorous laboratory
and instrumental conditions to improve
the stability of emission (78).
LA-ICP-MS is a recent, very effective
method to determine metals in food
samples. Since substantial part of the
sample preparation is avoided (there is
no preparation except that food is dried
at 110°C), when compared with classical
ICP-MS, this technique is faster, and
possible contamination effects due to
reagents is greatly limited. Moreover, it
allows accurate mapping of the analyzed
sample. However, due to kinetic effects in
laser ablation and/or in sample transfer to
ICP-MS system, elemental fractionation
is generated, so that results obtained by
LA-ICP-MS technique must be considered
semi-quantitative, unless the effects
of fractionation can be corrected by
instrumental calibration with adequate
standards (101,127,128).
Isotope dilution technique may be of very
high sensitivity (< 1 ppb), depending on
the isotopes and on their enrichment in
the tracer (spike) which is mixed to the
sample that must be analyzed. Moreover,
precision of the isotope dilution method
in determining the concentration of a
metal is related to the uncertainty which
afflicts the isotopic ratio of the element
which is measured for the technique.
This uncertainty is amplified by a sort of
magnification factor, the value of which
depends on sample/spike weight ratio.
Over- or underspiking of sample must
be carefully avoided because they may
determine magnification factor values
which are significantly higher than 1 (109112).
ID-MC-ICP-MS is greatly faster than
ID-MC-TIMS but the disadvantage is
the definitely lower precision both in
measuring isotopic ratios and in measuring
concentrations, so that ID-TIMS technique,
when possible, can be considered the most
effective method (61,122,129,130).
If metals must be determined at ultratrace
concentration levels (<1 ppm), particular
care should be taken in avoiding any
possible source of contamination during
sample preparation. At present, inductively
coupled plasma multi-collector mass
spectrometry and laser-ablation inductively
coupled plasma multi-collector mass
spectrometry, because of their dynamic
Monitoring Contaminants in Food Chain and their Impact on Human Health
range and capability for multi-element
analysis, are the most valuable methods
for the analysis of trace elements (78,83,
Investment and operational costs for ICPMS technique are however high, and are
not justified if a limited number of elements
must be determined for a limited number
of samples. In this case, AAS technique
or related will be preferred. Instead, for
the analysis of several elements in a large
numbers of samples the ICP-MS technique
is economically the most advantageous.
For this reason this technique is mainly
used in the most important government
analytical centres, where a large number
of elements in a variety of food matrices
are routinely determined (78,131,132).
4.2 Monitoring dioxins in food and in
biological matrices by high resolution
mass spectrometry
In recent years, based on a growing body
of evidence, there is an increasing concern
about the possible health threat posed
by substances present in environment,
food and consumer products termed
endocrine-disruptors (EDs) and defined as
“exogenous substances that cause adverse
health effects in an intact organism or
in its progeny, consequent to changes in
endocrine function” (133,134).
The group of molecules identified
as EDs is highly heterogeneous and
includes synthetic chemicals used as
industrial solvents/lubricants and their
by-products [polychlorinated biphenyls
(PBBs), dioxins], plastics [bisphenol
A (BPA)], plasticizers (phthalates),
pesticides [methoxychlor, chlorpyrifos,
dichlorodiphenylt r ichloroethane
(DDT)], fungicides (vinclozolin), and
pharmaceutical agents [diethylstilbestrol
(DES)]. Moreover, some naturally
occurring compounds, present in plants
and termed phytoestrogens, have been
found to posses estrogenic properties.
The majority of phytoestrogens belongs
to the large group of flavonoids. EDs
have long environmental half-life
resulting in a continue increase of their
global concentration in the environment;
furthermore, they have very low water
solubility and extremely high lipid
solubility, leading to their bioaccumulation
in adipose tissue. Although several
studies have definitively assess the toxic
properties of those polluting compounds,
conclusive evidences are still lacking on
the effect of low doses exposition and on
the synergistic effect of complex mixtures
of compounds. Different studies have been
performed in Germany(135), Belgium
(136), Sweden (33) and Japan (137), in
order to evaluate the body burden levels of
PCDDs/PCDFs and DL-PCBs on general
population. However, similar studies have
not been performed on general population
in Italy. The only data available up to now
for Italian population are those regarding
Seveso population who experienced the
highest levels of TCDD exposure known
in a residential population (138,139).
Concerning the chemical properties,
PCDDs, PCDFs and PCBs constitute a
group of 419 persistent environmental
chemicals. Only 17 congeners among
PCDDs and PCDFs and 12 congeners
among DL-PCBs cause toxic responses
similar to those caused by 2,3,7,8tetrachlorodibenzo-p-dioxin
the most toxic congener within these
groups of compounds. PCDDs, PCDFs,
and PCBs exist in environmental and
biological samples as complex mixtures
of various congeners with different rates
of degradation due to their different
solubility and volatility. Therefore, the
relative concentration of congeners differ
CNR Environment and Health Inter-departmental Project
across trophic levels and the composition
of these mixtures is often very different
from the one originally released into the
environment. The complex nature of these
chemicals complicates the health risk
evaluation for humans. In order to facilitate
risk assessment and regulatory control of
exposure to these mixtures the concept of
toxic equivalency factors (TEFs) has been
developed. TEF values are used to calculate
the toxic equivalent (TEQ) concentrations
in various matrices (animal tissues, soil,
sediment and water). TEFs and TEQs
are used for risk characterization and
management purposes (140,141).
In the frame of PIAS project, in a
tight collaboration with the other
groups participating to the project, we
propose to monitor the concentration
of polychlorinated dibenzo-p-dioxins
(PCDDs), dibenzofurans (PCDFs) and
dioxin like polychlorinated biphenyls
biphenyls (DL-PCBs) in blood samples
from non occupationally exposed subjects
as well as in selected food matrices,
typical of the Mediterranean diets such
as milk, mozzarella cheese, meat and
fish, which, being particularly rich in the
lipids, bio accumulate such molecules. For
the different matrices, the concentration
of some dioxins congeners (PCDDs,
PCDFs, PCBs) will be determined by
isotope dilution high resolution gas
chromatography/high resolution mass
spectrometry (HRGC/HRMS) (142-144).
Experimentally, after adding PCDD/F
and PCB congeners isotopically labelled
with the isotope 13Carbon, the samples
will be submitted to specific extraction
and gel clean-up steps. PCDD/F/PCB
concentration will be reported as pg/g fat,
pg WHO-TEQ/ g fat. The measurements
will be performed by means of HRGC/
HRMS: the mixtures will be separated
on a Gas Chromatographer, using a
DB-5 capillary column, coupled on line
with a high resolution double focusing
mass spectrometer. The Proteomic
and Biomolecular Mass Spectrometry
Center is equipped with a Autospec NT
instrument (Waters) specifically suitable
for the analysis of dioxins and dioxin-like
compounds, having a resolution higher
that 10.000 FWHM. The use of high
resolution capillary gas chromatography
and highly selective MS conditions (select
ion monitoring, resolution > 10.000,
accurate m/z assignment to 0.001 Da)
greatly reduces the potential for coextracted compounds to interfere with the
measurements of those analytes. Moreover,
the use of PCDD/F and PCB congeners
isotopically labelled allow the accurate
calculation of the analytes concentration.
However, it should be underlined that the
determination of dioxins concentration in
blood and food samples requires long and
laborious analytical procedures, high cost
of analysis (about 300 €/sample) as well
as the use of dedicated instruments and
highly expert operators.
The proposed biomonitoring will provide
results of relevant importance for the
estimation of the toxic human burden due
to both the environmental exposure and the
food chain. Information on whether and
what extent chemical substances are really
taken up from the environment (internal
dose) are of fundamental importance for
the evaluation of the related risk for human
health and to elucidate the effect of low
dose exposure.
4.3 Biomarkers to determine dietary
exposure to xenobiotics: the case of
cytochrome P450
The human cytochrome P450 (CYP)
superfamily, containing 57 genes (145),
contributes to the metabolism of a
variety of xenobiotics including drugs,
Monitoring Contaminants in Food Chain and their Impact on Human Health
carcinogens, constituents of food including
chemicals present as pollutants (146). The
resultant increases in polarity usually
facilitate excretion and are considered to
be a detoxification process, but in some
cases foreign compounds are converted
to products with much greater toxicity
(147). Chemicals present in the diet may
be metabolized by CYPs to non-toxic
metabolites and excreted, however the
formation of toxic metabolites is possible
(148). It was reported that xenobiotics may be
substrates, inhibitors or inducers of CYPs.
Natural products present in cruciferous
vegetables have been shown to selectively
up-regulate CYP1A1 and CYP1A2
isozymes on chronic ingestion (148). On
the other hand, several natural products
selectively inhibit mono-oxygenation,
especially in the intestine, and may lead
to increased bioavailability and reduced
metabolism of dietary components (149).
CYP1A is important as it is involved in
bioactivation of ubiquitous environmental
contaminants such as polychlorinated
dibenzo p-dioxines and in the past much
concern has been focused on the induction
of CYP1A as sensitive bioindicator for the
exposure of fish to these contaminants
in the marine environment (150). In this
context, we demonstrated that CYP1A can
be a useful probe for the exposure of adult
sea bass and frog to polycyclic aromatic
hydrocarbons (151). β-Naphtoflavone, a
typical polycyclic aromatic hydrocarbons
resulted in an induction in the liver
of CYP1A and the induction was
manifested by: i. immunoblot analysis
using anti-rat CYP1A1; ii. an increase
in CYP1A-mediated methoxyresorufinO-demethylase and ethoxyresorufin-Odeethylase activities.
We also demonstrated that the CYP2Alike inhibition can be used as biomarker
of exposure of herbicides, such as
dichlobenil. Expression of CYP2E1 in
human circulating lymphocytes has
raised clinical interest because it has been
proposed as a potential non-invasive bioassay determination of CYP2E1 expression
and activity in vivo (152). An elevation of
CYP2E1 has been reported in lymphocytes
from poorly controlled diabetic patients
(153). Considering that cytochrome P450
can be induced by several xenobiotics,
we can suppose that components of the
diet, mainly those present as pollutants or
additives, can modulate the cytochrome
P450 isoforms. For this reason we can use
this system as biomarker to assess dietary
exposure to xenobiotics. The studies
could be performed using animal models,
by the administration of extract of food
to evaluate modulation of CYPs. This
aspect can also be investigated in humans
using lymphocytes to assess if some CYP
isoforms are induced and/or inhibited
following ingestion of contaminants
present in the diet.
4.4 Marine invertebrate as model to assay
the effect of xenobiotics on reproduction
In the last decade, the international
scientific community has become
increasingly concerned that exposure
to low levels of synthetic chemicals or
xenobiotics may disturb hormone function
in man and animals (so-called endocrine
disruptors – Medical Research Council
(UK), 1995; Danish Environmental
Protection agency, 1995). During the
past 50 years, large quantities of diverse
xenobiotics have been released into the
environment as a consequence of efforts
to increase agricultural productivity and
as a result of modern manufacturing
processes and their by-products. These
chemicals include herbicides, pesticides,
fungicides, plasticizers, polystirenes,
PCBs, polychlorinated dibenzodioxidins,
CNR Environment and Health Inter-departmental Project
alkylphenolic compounds (154), organotins,
and more specifically tributyltins (TBT),
used for its biocide properties as the
active agent in antifouling paints (155).
There is increasing evidence that these
xenobiotics in the environment may
disrupt the endocrine systems of aquatic
life and wildlife. In addition, EDs and
other food-contaminating environmental
pollutants represent a high risk factors
in animal reproduction (156). Such
chemicals are receiving more and more
attention, particularly because several
compounds not specifically designed to
possess endocrine activity have been
shown to possess unexpected hormonal
activity in a wide variety of organisms.
Reproductive hormone-receptor systems
appear to be especially vulnerable; in fact,
some EDs can interfere with the normal
mechanisms of steroid hormone action and
with the embryonic development of the
male and female reproductive systems of
wildlife and experimental animals which
in turn may affect normal reproductive
functions in adulthood (154). It has been
demonstrated that xenobiotics acting
through steroid-dependent mechanisms,
interact with estrogen receptors, androgen
receptors, or with certain steroid binding
proteins (ABP, SHBG). The endocrine and
reproductive effects of EDs are believed to
be due to their ability of: i. mimicking the
effects of hormones; ii. altering the pattern
of synthesis and metabolism of hormones;
iii. antagonizing the effects of hormones; iv.
modifying hormone-receptor levels (157).
In general, the magnitude of the cellular
response to hormones is dependent upon
the number of receptors occupied by the
hormone which in turn is related to hormone
concentration. Therefore, EDs could
potentially alter endocrine functions by
influencing the concentration of hormones
through changes in the rates of their
secretion or metabolism, or by interfering
with hormone action at the receptor or
at other sites along the hormone signal
transduction pathway. It is well known
that many aspects of the reproduction
in vertebrates are under the control of
hormones and sex steroids and a great
deal of evidence has been accumulating,
showing that it may also be the case in
invertebrates and fish (158-161). Several
types of sex steroids have been detected in
various species of invertebrates (162-164).
Estradiol-17β and progesterone have been
found in the tissue and hemolymph of the
American lobster (165,166). In Paeneus
monodon estradiol-17β and progesterone
levels in the hemolymph, ovaries and
hepatopancreas were related to the ovarian
stage of development (167). Injections of
progesterone and 17αOH progesterone
induce ovarian maturation in Metapeneus
ensis (168) and stimulate vitellogenin
secretion in Paeneus japonicas (169)
Estrogens stimulate vitellogenin synthesis
in Macrobrachium rosenbergii and
in Paeneus monodon (167,170). In the
female of Pandalus kessleri the level of
estradiol coincides with vitellogenin in
the hemolymph (166). In P. monodon
estrogen treatment during vitellogenesis
may suppress molting, while stimulating
vitellogenin production (167). Sex steroid
hormones (androgens, progesterone,
estradiol-17β) and 3 β-hydroxysteroid
deydrogenase, a key enzyme in
steroidogenesis, have been reported in
the gonad of the male of the cephalopod
Octopus vulgaris (171-174).
Many animal models are suitable for
comparative studies with mammalian
models in particular the marine invertebrate
Ciona intestinalis (ascidians) share many
common biological mechanisms with
vertebrates (175). The effect of compounds
deriving from marine diatoms have been
Monitoring Contaminants in Food Chain and their Impact on Human Health
already investigated showing an influence
at molecular level on the initial mechanism
of fertilization. This effect seems also
to influence the following embryo
development (176). Similarly to ascidians,
also the mollusk Octopus vulgaris
share basic biological mechanisms with
mammals. The germinal vesicle breakdown
which is the first event in oocyte maturation
appears to be supported by an ion current
activity of specific L-type calcium
channels occurring also in ascidians and
mammals (177-179). At present, a study is
in progress on the effects of four different
heavy metals lead, cadmium, zinc and
copper on the ion currents present on
the plasma membrane of the oocytes
and on the electrical events involved in
the processes of maturation fertilization
and embryo development of the ascidian
Ciona intestinalis. Data obtained show
an inhibition of either plasma membrane
currents and the first events of fertilization.
These results suggest a plausible negative
impact of the xenobiotics on the early
events of reproduction in model animals.
environmental chemicals to exert toxicity
on human health and reproductive fitness
remains largely speculative, evidence are
accumulating that multiple stressors from
contaminated environment may adversely
affect populations of marine animals and
mammals such as humans by interfering
with similar known processes of the
reproductive process (58,180).
Specific goals of PIAS are to propose new
projects at national and European level
which may fill some of the gaps in the
literature regarding the complex interaction
between contaminants and human health.
This working group identified the need
to determine a real cause-effect relation
between level of contaminants in the diet,
their “real” presence in selected human
populations and their effect on health. In
current scientific literature, this problem
has been successfully approached with
excellent studies where measurements of
contaminants present in the environment
and bio-accumulated in the food chain have
been linked to individual consumption
extrapolating, from these data, the human
intake in specific age groups. As already
mentioned above (section 2.2), a significant
example comes from the study of Bilau
and co-workers (29) who report data on the
dietary exposure to dioxin-like compounds
in adolescents, their mothers and adults,
a result of the Flemish Environment and
Health Study (www.milieu-en-gezondheid.
be). They demonstrated that in the selected
aged groups, the median (95th percentile)
estimated daily intakes of dioxin-like
contaminants were 2.24 (4.61), 2.09
(4.26), and 1.74 (3.53) pg CALUXTEQ
kg-1bwd-1 for, respectively, adolescents,
mothers and adults. These values exceed
the tolerable weekly intake (TWI) of 14
pg WHO-TEQ kg-1bww-1, as derived by
the Scientific Committee on Food (181).
The relative validity and reproducibility of
this experimental approach was assessed
by the same authors in a different study
(182). Here, they concluded that the
food frequency questionnaire designed
to estimate the intake of dioxin-like
contaminants represents a valuable tool
for ranking individuals in the study
population on the basis of estimated intake
of dioxin-like contaminants. However,
absolute intakes should be estimated
without correction factors and interpreted
with caution. In a similar study carried
CNR Environment and Health Inter-departmental Project
out by a Swedish group within the EU
funded CASCADE Network of Excellence
(Contract N. FOOD-CT-2003-506319) the
dietary intake of dioxin-like pollutants
was investigated in children and young
adults (33). The results showed that among
the selected Swedish population, boys
and girls up to the age of ten years had
a median TEQ intake that exceeded the
tolerable daily intake (TDI) of 2 pg TEQ/
kg body weight. Dairy and fish products
were the main sources of exposure. In fact,
the individuals most highly exposed were
characterized by a high consumption of fish.
Also in this case, exposure was estimated
matching the concentration data of dioxinlike compound in food commodities
(meat, fish, dairy products, egg, edible fats
and other foodstuff) with food intake data.
Similar studies on dioxins exposure via
food were performed in several countries,
generally showing that estimated dietary
intake is above the recommended TDI level
ranging from roughly 2–6 pg TEQ/kg bw/
day (183-192). On the opposite, an Italian
study established that the mean value of
dioxins measured in food of animal origin
by isotope dilution method was 0.144 ±
0.266 pg-TEQ/g (range: 0.003–1.655 pgTEQ/g). The average daily food intake was
obtained from national data collected by the
National Institute of Nutrition, and from a
cohort study on diet and cancer including
40,000 Italian subjects. The conclusion
was that the major contribution to dioxins
intake with food comes from cow milk
and fish consumption and were below the
limits set by the European legislation (193).
Apparently, the adherence to the limits
established by EC (194) was confirmed
in parallel studies carried out in different
Countries such as Germany (195), Finland
(196), Japan (197) and Spain (198).
As discussed by others (193), many of
the studies cited suffer from the same
limitations: i. the amount of dioxins,
or other contaminants, intake with diet
was estimated from national surveys or
epidemiological studies, without measuring
dioxins content of a certain food and the
individual intake of that foodstuff; ii.
data were obtained through a monitoring
program, not as part of a research project;
this means that the aim of the monitoring
was not to study human exposure through
food, but to assess food content of dioxins
and other residues; iii. dioxin content was
lacking for certain food consumed within
the population, making the analyses
Based on this preamble, we proposed
within the PIAS project a large, multicenter
and multidisciplinary study with an
extended follow-up which will take into
account the missing information existing
in the Literature. The target population
will be represented by children and young
adults. In fact, they constitute a vulnerable
group and previous studies suggest that
it is essential to perform age-specific
dietary intake assessments to more
carefully consider, in the risk management
processes, sensitive and/or highly exposed
individuals in the population.
The general objective of this proposal is:
to address the healthy status of a young
population determining the concentration
of xenobiotics which come to humans
through the food chain.
The proposal consists in two phases:
Phase I
The working hypothesis is illustrated in
Fig. 3. The experimental aim is to study
populations of children, adolescents and
adults living in various Italian regions,
including Campania (South Italy)
establishing biological banks (mainly blood
and urine samples). These individuals
will be selected from large Italian cohorts
that are at least in part already available
Monitoring Contaminants in Food Chain and their Impact on Human Health
from ongoing European projects involving
members of PIAS working groups. The
Italian study sample should be composed
by about 2000 individuals with males and
females equally represented. Additional
cohorts with similar features from
other Countries potentially interested to
participate will be enrolled in the study
to obtain comparable information at
European level. At baseline the following
variables will be measured: in complete
anthropometry including body composition
including selected hormones; physical
activity tests; medical history, behavioral
and socio-economic questionnaires; foodfrequency questionnaire and repeated 24h
dietary record. All these variables will be
measured again in the same population in
the follow-up survey.
Biological samples, preferentially blood
and urines, freshly collected or, where
possible, already available if conveniently
stored for the expected analyses, will
be employed to measure the presence of
those contaminants whose presence was
independently and previously verified
in food commodities taken directly
from or through the food chain. A
careful evaluation of different types of
contaminants/xenobiotics to be analyzed
in the present project is actually under
scrutiny from members of PIAS working
groups. The selection will certainly include
compounds belonging to the following
categories: pesticides, dioxins and dioxinlike molecules and heavy metals. For the
choice, two main criteria will be followed:
i. presence of these compounds in the diet
of the selected individuals; ii. availability
of official methods to detect them in
biological samples and foods.
At the end of this phase of the study,
we will relate the level of contaminants
present in foods and biological samples
with epidemiological data from the
populations under study (dietary habits,
health status) to determine the potential
association between concentrations of
selected contaminants and health effects.
General methods. In order to assess the
exposure in children and young adults,
the individuals will be stratified by gender
and age. Individuals with incomplete
information on body weight or food
consumption will be excluded. The 100
most commonly consumed food items
will be collected and analyzed by standard
methodology to assess the presence and
concentration of selected contaminants.
Food items will be obtained from producers
or purchased from different stores in the
cities where the cohorts will be recruited.
Accordingly to reports periodically
published by EC (199-201), the food
groups chosen for the study will be: i. fish,
dairy products, egg, edible fats and other
fat-containing products for the presence of
dioxins, dioxin-like molecules and selected
metals; ii. cereals, fruits, vegetables,
beverages, vegetable soups and sugar for the
presence of pesticides, biocides and heavy
metals. Exposure to different xenobiotics
based on consumption of various food
items by each individual will be expressed
accordingly to international units establish
for each specific contaminants. Data will be
analyzed by standard statistical methods.
In particular, dioxins concentration in
different biological samples and in food
groups will be associated with selected
health outcomes after adjustment for age
and gender. The dioxin levels in different
food groups (fish, meat, dairy products, egg,
edible fats, other fat-containing products,
fruits, vegetables, cereals, etc.) will be
compared and related to the individual
intake of each food item to assess the
principal sources of exposure both at the
individual and at the population level. The
CNR Environment and Health Inter-departmental Project
proposed sample size of 2000 individuals is
large enough to allow statistical power for
different series of analysis. For instance, it
will allow to detect a 5% difference in the
amount of ingested contaminants between
groups, at alpha =0.05 and beta=80%.
Phase II
Information obtained from doses of
contaminants in biological samples
and above the threshold established
by international health agencies will
represent the starting point for phase II
of the proposal devoted to determine
whether and how, from a molecular point
of view, exposure to these xenobiotics may
interfere with the normal physiological
state of the cell/organism resulting in
pathological conditions in adults. As
schematically represented in Fig. 3, phase
II will take advantage of different cellular
and animal experimental models suitable
to address cause-effect relation in specific
chronic diseases potentially associated to
exposure and/or accumulation of dietary
contaminants. Decision about the diseases
on which to focus our attention will strictly
depend upon results obtained after phase I.
In choosing the research groups to assign
these specific tasks, we will primarily
consider expertise and competence
within CNR, as resulted from the census
questionnaires settled down during the
course of PIAS program (see section 3.2
above). Phase II will also takes advantage
of the work and competence deriving from
other working groups within PIAS (e.g,
endocrine disruptors). As an example of
activity performed during phase II of the
project, great importance will be devoted
to assess the potential effect of xenobiotic
exposure to reproductive fitness and
development (see section 4.4) and to the
role of cytochrome P450 in metabolizing
xenobiotics (see section 4.3).
Key issues to be addressed in order to
correctly evaluate and interpret data
deriving from the experimental models
employed in phase II concern: i. genetic
background; ii. adaptive response/
metabolism of different xenobiotics.
Risk estimates routinely reflect numerous
sources of both uncertainty (which describes
the range of plausible risk estimates arising
because of limitations in knowledge) and
variability (which describes the range of
risks arising because of true differences).
Among them, and besides age and gender,
genetic differences among members of
the population may play a relevant role.
Since the majority of the study on dioxins
have been conducted using genetically
homogeneous inbred mice to characterize
the risk, their conclusions should be
taken very cautiously when applied to the
genetically variable human population.
Although well-designed occupational and
environmental epidemiological studies
can yield useful information on human
population variability, relatively little
quantitative information is available
about the potential impact on genetic
polymorphisms in the human population
that might give rise to differences in
susceptibility to the toxic effects of dioxins,
and DLCs. As an example of candidate
gene, the Aryl hydrocarbon receptor (AhR
or AHR) is a cytosolic transcription factor
able to bind to chemicals such as TCDD,
leading to changes in gene transcription.
A state of the art revision of the literature
will be done in the course of this project
to identify candidate genes or biological
pathways to be explored with genetic
studies. The panels of SNPs in selected
genes will be genotyped in the laboratories
of ISA-CNR, using up-to-date genotyping
technologies. At the current status, allelic
discrimination will be performed by
TaqMan® genotyping assay. The use of
Monitoring Contaminants in Food Chain and their Impact on Human Health
other techniques like SNPs array will
be considered taking into account the
number of samples/SNPs to be evaluated
and the cost of the assay at the time of
genotyping. The genotyped SNPs will be
uploaded in a central database and linked
to the phenotype data. To minimize the
population stratification bias, a potential
source of false positive associations in
genetic population studies (202), genetic
analyses will be restricted to individuals
of Caucasian origin. Additionally, it is
likely that all the cohorts will belong
to single countries, with the majority
of participants coming from specific
delimited geographical areas, thus further
reducing the risk of population admixture.
Hormesis is a biphasic dose-response
phenomenon characterized by a low-dose
stimulation and a high-dose inhibition
resulting in a U- or inverted U-shaped
dose response (203). The phenomenon of
biphasic dose-response relationship has
received considerable attention over the
past few years (204). A good example on
the application of hormesis phenomenon
to human health derives from exposure
to heavy metals, such as lead, cadmium,
mercury and arsenic. Cadmium is a potent
carcinogen in a number of tissues, and is
classified by IARC as a human carcinogen
(205). Reactive oxygen species (ROS) are
often implicated in cadmium toxicology,
either in a variety of cell culture systems
(206-209), or in intact animals through all
routes of exposure (210-213). However,
in contrast to acute toxicity, the roles of
ROS in chronic cadmium toxicity and
carcinogenesis have been controversial
depending on experimental conditions.
On the other hand, administration of
cadmium to animals at low levels for
one year increases hepatic and renal
glutathione levels, without elevations
in tissue lipid peroxidation levels (214).
A biphasic ROS response to cadmium
exposure through the drinking water has
also been proposed. ROS and ROS-related
gene expression occur right after cadmium
exposure, but return to normal levels after
8 weeks of exposure (215). A further
example comes from chromium (VI),
a well-known mutagen and carcinogen
that produces ROS during formation of
reactive chromium intermediates (216218) and induces oxidative stress (219).
ROS are known to be generated in various
cell types, such as K562 leukemic cells,
J774A.1 murine macrophages (220) and
human epithelial like L-41 cells (221) when
acutely exposed to chromium (VI). A
potential adaptive response were obtained
when immortalized rat osteoblasts (FFC
cell line) and U937 were exposed to 0.050.5 micromolar chromium (VI) for 4 weeks
(222). In addition proteomic analysis of
both FFC and U937 cells exposed to 0.5
micromolar chromium (VI) resulted in a
differential time dependent regulation of
glycolytic, stress and cytoskeletal proteins
that play an essential role in normal cellular
functioning such as energy metabolism,
cell signaling and proliferation (223).
Hormetic responses to xenobiotic
exposure likely occurring as a result of
overcompensation by the homeostatic
control systems operating in biological
organisms have been discussed in excellent
reviews which commented on the economic
implications of hormesis (203,224-231).
Bioavailability refers to the extent to
which humans and ecological receptors
are exposed to contaminants from soil,
or sediment directly or through the
food chain (an extensive and excellent
review on this topic has been published
by the Committee on Bioavailability of
Contaminants in Soils and Sediments of
the National Research Council)(232). Our
interest in determining the bioavailability
CNR Environment and Health Inter-departmental Project
of potentially contaminants adsorbed with
the diet is related to their risk assessment
for human health. However, the concept
of bioavailability has recently been
exploited by hazardous waste industry as
an important consideration in deciding
how much waste can be left in place
without creating additional risk if these
contaminants are not bioavailable (232).
In terms of effects on human physiology,
once absorbed, contaminants may be
metabolized, excreted, or they may cause
toxic effects. Several levels of uncertainty
are associated with bioavailability,
including: i. limited knowledge about how
biota modify bioavailability of chemicals
which come into contact with digestive
systems, and whether information
obtained for one species is representative
of others; ii. the effect of food processing
(cocking, pressing, etc.) which makes
extremely difficult to precisely calculate
the daily intake of xenobiotics, despite
the nominal values determined in the
single food component; iii. the synergic
or antagonistic effect on absorption and
metabolism of xenobiotics by other food
components, such as polyphenols. In this
contest, literature reports examples in both
directions: the protective effect of oral
resveratrol on the sub-acute toxic effects
of TCDD in C57BL/6J mice (233), or the
confounding activity of contaminating
metals which may interfere with the
regulated absorption, distribution, and
excretion kinetics of essential metals (234),
although this study conclude that food
contaminations with metals are too low
to have an impact on the bioavailability
of essential metals. In general, a higher
priority could be given to studies exploring
combinations of nutrients, xenobiotics and
food contaminants, at realistic intestinal
concentrations, with hazardous or
beneficial impacts on human health using
high throughput in vitro tools (235).
“We are what we eat,” says Ayurveda, the
ancient Indian science of life. This sentence
immediately clarifies the perception of
how food can influence our lives and the
relevance of key issues as food safety
on human health. In this Chapter, we
reviewed critical aspects concerning food
contamination and its impact on health.
Our analyses showed that several gaps
and uncertainties remain, despite the great
concern for “food integrity” and the variety
of scientific contributions from different
fields, such as environmental science, food
chemistry, human epidemiology. These
limits cannot be bypassed only with the
efforts of the scientific community, but
the contribution of Governments and
international environmental and healthy
agencies is mandatory to face and rapidly
solve these deficiencies. This is a critical
issue: in evaluating food safety, no gaps
are allowed since bad or incomplete
information may result in an enormous
hazard for consumer’s safety and for the
negative consequences that the presence
of a contamination in the food chain may
generate to the social and economical life
of a region.
To correctly approach the issue of the
negative influence (if any) of chemical
contaminants reaching people throughout
diet, we underlined the need of an
integrated approach which considers key
factors, such as the genetic background of
exposed subjects, adaptive responses, age
dependent accumulation and bioavailability
of specific compounds. Very recently,
the publication of the new European
regulation concerning the use of pesticides
in agriculture (17), raised the problem of
the relation existing between exposure to
Monitoring Contaminants in Food Chain and their Impact on Human Health
Figure 3: Schematic model of research program proposed within PIAS
low dose contaminants and their potential
harmful effects in developing chronic
pathologies. Several validated experimental
models exist to study the consequences on
human health of acute exposure to elevated
concentration of chemical contaminants.
On the opposite, a general consensus is
still far to be reached on how to assess the
effect of prolonged, low-doses exposure
to environmental contaminants. Perhaps,
a lessons may derive from studies in the
filed of radiation exposure.
In this context, PIAS proposed a large,
multidisciplinary research project aimed to
fill, at least in part, the lack of information
existing in the field. The realization of
this study cannot be obtained solely by
expertise already present within the CNR,
but requires the strong contribution of
national and European groups with proved
experience in the numerous and different
fields considered by the proposal (Fig. 3).
The realization of an integrated approach
to assess the impact of food contamination
on human health represents, in our view,
the only correct way to increase scientific
knowledge and build trust and confidence
in the consumers’ beliefs.
KEYWORDS: heavy metals, pesticides,
dioxins, xenobiotics, food chain, human
health, bioavailability, hormesis.
Gove PB. Webster’s third new international
dictionary. Springfield, Mass, MerriamWebster Inc, 1993.
Kogevinas M. Human health effects
of dioxins: cancer, reproductive and
endocrine system effects. Hum Reprod
Update 2001; 7: 331-339.
Stefanidou M, Maravelias C, Spiliopoulou
CNR Environment and Health Inter-departmental Project
C. Human exposure to endocrine disruptors
and breast milk. Endocr Metab Immune
Disord Drug Targets 2009; 9: 269-276.
WWF-UK. ContamiNATION, the results
of WWF’s biomonitoring survey 2003.
Hayashi Y. Scientific basis for risk
analysis of food-related substances with
particular reference to health effects on
children. J Toxicol Sci 2009; 34 Suppl 2:
IFT Expert Report. Making Decisions
about the Risks of Chemicals in Foods
with Limited Scientific Information.
Comprehensive reviews in food science
and food safety 2009; 8: 269-303.
La Rocca C, Mantovani A. From
environment to food: the case of PCB.
Ann Ist Super Sanità 2006; 42: 410-416.
Hu H. Human health and heavy metals
exposure. MIT press, 2002.
Rangan C, Barceloux D. Food
contamination. Hoboken, NJ, John Wiley
& Sons, 2008.
Environmental Protection Agency. www.
Grandjean P, Weihe P, White RF, Debes
F. Cognitive performance of children
prenatally exposed to “safe” levels of
methylmercury. Environ Res 1998; 77:
Murata K, Weihe P, Araki S, BudtzJorgensen E, Grandjean P. Evoked
potentials in Faroese children prenatally
exposed to methylmercury. Neurotoxicol
Teratol 1999; 21: 471-472.
Col M, Col C, Soran A, Sayli BS, Ozturk
S. Arsenic-related Bowen’s disease,
palmar keratosis, and skin cancer. Environ
Health Perspect 1999; 107: 687-689.
Villarejo D, McCurdy SA. The California
Agricultural Workers Health Survey. J
Agric Saf Health 2008; 14: 135-146.
Balali-Mood M, Balali-Mood K.
Neurotoxic disorders of organophosphorus
compounds and their managements. Arch
Iran Med 2008; 11: 65-89.
Steenland K, Cedillo L, Tucker J et
al. Thyroid hormones and cytogenetic
outcomes in backpack sprayers using
fungicides in Mexico. Environ Health
Perspect 1997; 105: 1126-1130.
Commission E. Regulation (EC) No
1107/2009 of the European Parliament
and of the Council of 21 October 2009
concerning the placing of plant protection
products on the market and repealing
Council Directives 79/117/EEC and
91/414/EEC. 2009.
Baccarelli A, Pesatori AC, Masten SA et
al. Aryl-hydrocarbon receptor-dependent
pathway and toxic effects of TCDD in
humans: a population-based study in
Seveso, Italy. Toxicol Lett 2004; 149: 287293.
Adamsson A, Simanainen U, Viluksela
M, Paranko J, Toppari J. The effects of
2,3,7,8-tetrachlorodibenzo-p-dioxin on
foetal male rat steroidogenesis. Int J
Androl 2009; 32: 575-585.
Cao Y, Winneke G, Wilhelm M et al.
Environmental exposure to dioxins and
polychlorinated biphenyls reduce levels
of gonadal hormones in newborns: results
from the Duisburg cohort study. Int J Hyg
Environ Health 2008; 211: 30-39.
Amerine M, Pangborn R, Roessler E.
Principles of Sensory Evaluation of
Foods. Academic press, 1965, pp.
Cardello A. Food quality: relativity,
context and consumer expectations. Food
quality and preferences 1965; 6: 163-170.
Adu-Amankwa P. Quality and process
control in the food industry. The Ghana
Engineer; 1999; 1999.
Nin J. New technology for food systems
and security. Asia Pac J Clin Nutr 2009;
18: 546-548.
Karlen D, Mausbach M, Doran J, Cline R,
Harris R, Schuman G. Soil quality: a concept,
definition and framework for evaluation. Soil
Sci Soc Am J 1997; 61: 4-10.
Henry J. Processing, manufacturing, uses
and labelling of fats in the food supply.
Ann Nutr Metab 2009; 55: 273-300.
Lau OW, Wong SK. Contamination in food
from packaging material. J Chromatogr
A 2000; 882: 255-270.
Thacker SB, Stroup DF, Parrish
RG, Anderson HA. Surveillance in
Monitoring Contaminants in Food Chain and their Impact on Human Health
environmental public health: issues,
systems, and sources. Am J Public Health
1996; 86: 633-638.
Bilau M, Matthys C, Baeyens W et
al. Dietary exposure to dioxin-like
compounds in three age groups: results
from the Flemish environment and health
study. Chemosphere 2008; 70: 584-592.
Willett WC. Future directions in
the development of food-frequency
questionnaires. Am J Clin Nutr 1994; 59:
Vanderperren H, Van Wouwe N, Behets
S, Windal I, Van Overmeire I, Fontaine
A. TEQ-value determinations of animal
feed; emphasis on the CALUX bioassay
validation. Talanta 2004; 63: 1277-1280.
Lambe J. The use of food consumption
data in assessments of exposure to food
chemicals including the application of
probabilistic modelling. Proc Nutr Soc
2002; 61: 11-18.
Bergkvist C, Oberg M, Appelgren M et
al. Exposure to dioxin-like pollutants via
different food commodities in Swedish
children and young adults. Food Chem
Toxicol 2008; 46; 3360-3367.
Rapid Alert System for Food and Feed
Beaton GH. Criteria of an adequate diet.
Philadelphia, Lea and Febiger, 1994.
Smith JC, Jr, Anderson RA, Ferretti R et
al. Evaluation of published data pertaining
to mineral composition of human tissue.
Fed Proc 1981; 40: 2120-2125.
Versiek J, Cornelis R. Normal levels of
trace elements in human blood plasma
and serum. Analytica chimica acta 1980;
116: 217-254.
WHO. Diet nutrition and prevention
of chronic disease. Technical Report
Series No. 797. Geneva: World Health
Organization; 1990.
Wolf WR. Biological reference materials:
availability, uses, and need for variation
of nutrient measurement. New York,
John Wiley, 1985.
Food and Nutrition Board Recommended
Dietary Allowances. Washington D.C, 1989.
41. Barnes DG, Dourson M. Reference dose
(RfD): description and use in health risk
assessments. Regul Toxicol Pharmacol
1988; 8: 471-486.
42. Black AL. Setting acceptance levels of
contaminants. Proceedings of the Nutrition
Society of Australia 1992; 17: 36-41.
43. Bolger PM, Yess NJ, Gunderson EL,
Troxell TC, Carrington CD. Identification
and reduction of sources of dietary lead
in the United States. Food Addit Contam
1996; 13: 53-60.
44. Hatchcock J. Safety evaluation of
vitamins and minerals. Chichester (UK),
John Wiley, 1998.
45. Hathcock JN. Safety limits for nutrient
intakes: concepts and data requirements.
Nutr Rev 1993; 51:278-285.
46. McLaughlin MJ, Parker DR, Clarke JM.
Metals and micronutrients – food safety
issues. Field Crops Research 1999; 60:
47. Solgaard P, Arkrog A, Fenger J, Flyger H,
Graabaek AM. Lead in Danish food-stuffs.
Evidence of decreasing concentrations.
Dan Med Bull 1979; 26: 179-182.
48. Ybanez N, Montoro R. Trace element
food toxicology: an old and ever-growing
discipline. Crit Rev Food Sci Nutr 1996;
36: 299-320.
49. Baldwin DR, Marshall WJ. Heavy metal
poisoning and its laboratory investigation.
Ann Clin Biochem 1999; 36: 267-300.
50. Russel LH. Heavy metals in foods of
animal origin. New York, 1978.
51. Alloway B. Heavy metals in soil. London,
52. Lisk DJ. Trace metals in soils, plants and
animals. Advances in Agronomy 1972;
24: 267-320.
53. Berrow MI, Webber J. The use of sewage
sludge in agriculture. Journal of the
Science of Food and Agriculture 1972;
23: 93-100.
54. Cox PA. The elements on Earth: Inorganic
Chemistry in the Environment. Oxford,
Oxford University press, 1995.
55. McBride MB. Environmental Chemistry
of Soils. Oxford, Oxford University
press, 1994.
CNR Environment and Health Inter-departmental Project
56. Panteeva SV, Gladkochoub DP, Donskaya
TV, Markova VV, G.P. S. Determination
of 24 trace elements in felsic rocks
by inductively coupled plasma mass
spectrometry after lithium metaborate
fusion. Spectrochimica Acta B 2003; 58:
57. Plant J, Smith D, Williams L.
Environmmental geochemistry at the
global scale. Journ geol Soc of London
2000; 157: 837-849.
58. Potts PJ. Geoanalysis: Past, Present and
Future. Analyst 1997; 122: 1179-1186.
59. Sparks DL. Environmental Soil Chemistry.
San Diego, Academic press, 1995.
60. Strenstrom T, Vahter M. Heavy metals
in sewage sludge for use on agricultural
soils. Ambio 1974; 3: 91-92.
61. Bennett-Chambers M, Davies P, Knott
B. Cadmium in aquatic ecosystems in
Western Australia. A legacy of nutrientdeficient soils. Journal of Environmental
Management 1999; 57: 283-295.
62. Fortunato G, Mumic K, Wunderli
S, Pillonel L, Bosset JO, Gremaud
G. Application of strontium isotope
abundance ratios measured by MC-ICPMS for food authentication. Journal of
Analytical Atomic Spectroscopy 2004;
19: 227-234.
63. Kelly S, Heaton K, Hoogewerff J. Tracing
the geographical origin of food: the
application of multi-element and multiisotope analysis. Trends in Food Science
and Technology 2005; 16: 555-567.
64. Kornexl BE, Werner T, Rossmann A,
Schmidt HL. Measurements of stable
isotope abundances in milk and milk
ingredients – a possible tool for origin
assignment and quality control. Z Lebensm
Unters Forsch A 1997; 205: 19-24.
65. Manca G, Camin F, Coloru GC et al.
Characterization of the geographical
origin of Pecorino Sardo cheese by casein
stable isotope ((13)c/(12)c and (15)n/(14)n)
ratios and free amino acid ratios. J Agric
Food Chem 2001; 49;:1404-1409.
66. Manca G, Franco MA, Versini G, Camin F,
Rossmann A, Tola A. Correlation between
multielement stable isotope ratio and
geographical origin in Peretta cows’ milk
cheese. J Dairy Sci 2006; 89: 831-839.
Pillonel L, Badertscher R, Casey M, Meyer
J, Rossmann A, al S-CHe. Geographic
origin of European Emmenthal cheese:
Characterisation and descriptive statistics.
International Dairy Journal 2005; 15: 547556.
Pillonel L, Badertscher R, Froidevaux P,
Haberhauer G, Holzl S, Horn Pea. Stable
isotope ratios, major, trace and radioactive
elements in emmental cheeses of different
origins. Lebensmittel-Wissenschaft undTechnologie 2003; 36: 615-623.
Varo P, Koivistoinen P. Mineral element
composition of Finnish food.XII General
discussion and nutritional evaluation.
Acta Agricultural Sacndinavica 1980; 22:
Williams CH, David DJ. Heavy metals
in Australian soils. Australian Journal of
Soil Research 1973; 11: 43-50.
Booth CK, Reilly C, Farmakalidis E.
Mineral composition of Australian readyto-eat breakfast cereals. Journal of Food
composition and Analysis 1996; 9: 135-147.
Borocz-Szabo M. Effects of metals
on sensory qualities of food. Acta
Alimentaria 1980; 9: 341-356.
Kanner J. Oxidative processes in meat
and products: quality implications. Meat
Science 1994; 36: 169-189.
Phillips LG, Barbano DM. The influence
of fat substitutes based on protein and
titanium dioxide an the sensory properties
of low fat milks. Journal of Dairy Science
1997; 80.
Semwal AD, Murthy MCN, S.S A. Metal
contents in some of the processed foods
and their effects on the storage stability of
precooked dehydrated flaked Bengalgram
Dahl. Journal of Fodd Science and
Technology – Mysore 1995; 32: 386-390.
Barnes KW. A streamlined approach to the
determination of trace elements in food.
Atomic Spectroscopy 1998; 19: 31-39.
Blyth AW. Foods: their Composition and
Analysis: A Manual for the Use of Analytical
Chemists and Others. London, 1986.
Brown RJC, Milton MJT. Analytical
Monitoring Contaminants in Food Chain and their Impact on Human Health
techniques for trace element analysis: an
overview. Trends in analytical Chemistry
2005; 24: 266-274.
Caroli S. The determination of chemical
elements in food. Applications for atomic and
mass spectrometry. Hoboken (NJ), 2007.
Sorin M, Cosnier A. Application of
ICP-OES to the Analysis of Food and
Agriculture: turkey, pork, hay and
soy samples. ICP atomic emission
spectroscopy Application note 38.
Akter KA, Chen Z, Smith L, Davey D,
Naidu R. Speciation of arsenic in ground
water samples: a comparative study of
Talanta 2005; 68: 406-415.
Herrera MC, Luque de Castro MD.
Dynamic approach based on iterative
change of the flow direction for
microwave-assisted leaching of cadmium
and lead from plant prior to GF-AAS. J
Anal At Spectrom 2002; 378: 1376-1381.
Hirano S, Suzuki KT. Exposure,
metabolism, and toxicity of rare earths
and related compounds. Environ Health
Perspect 1996; 104 Suppl 1: 85-95.
Priego-Capote F, Luque de Castro MD.
Dynamic ultrasound-assisted leaching of
essential macro and micronutrient metal
elements from animal feeds prior to flame
atomic absorption spectrometry. Anal
Bioanal Chem 2004; 378: 1376-1381.
Xiu_Ping Y, Yan L, Yan J. A flow
injection on-line displacement/sorption
preconcentration and separation technique
coupled with flame atomic absorption
spectrometry for the determination of
trace copper in complicated matrices.
JAnalAt Spectrom 2002; 17: 610-615.
Yebra mC, Carro N, Enriquez MF,
Moreno-Ciad A, Garcia A. Field sample
preconcentration of copper in sea water
using chelating minicolumns subsequently
incorporated on a flow-injection-flame
atomic absorption spectrometry system.
Analyst 2001; 126: 933-937.
Gomez-Ariza JL, Garcia-Barrera T,
Lorenzo F, Bernal V, Villegas MJ,
Oliveira V. Use of mass spectrometry
techniques for the characterization of
metal bound to proteins (metallomics) in
biological systems. Rev Anal Chim Acta
2004; 524: 15-22.
Kosters J, Diaz-Bonea RA, PlanerFriedrich B, Rothweiler B, Hirner AV.
Identification of organic arsenic, tin,
antimony and tellurium compounds in
environmental samples by GC-MS. J Mol
Structure 2003; 661-662: 347-356.
Grosser AZ, Neubauer K, Thompson L,
Davidowski L. A Comparison of ICPOES and ICP-MS for the Determination
of Metals in Food. Advanstar Publication,
Grotti M, Magi E, Frache R. Multivariate
investigation of matrix effects in
inductively couplet plasma atomic
emission spectrometry using pneumatic
or ultrasonic nebulization. J Anal At
Spectrom 2000; 15: 89-95.
Karami H, Mousavi MF, Yamini Y,
Shamsipur M. On-line preconcentration
and simultaneous determination of heavy
metal ions by inductively couplet plasmaatomic emission spectrometry. Anal
Chim Acta 2004; 509: 89-94.
Beauchemin D, Kyser K, Chipley
D. Inductively coupled plasma mass
spectrometry with on-line leaching:
a method to assess the mobility and
fractionation of elements. Anal Chem
2002; 74: 3924-3928.
Becker JS, Sela H, Dobrowolska J, Zoriy
M, Becker JS. Recent application on
isotope ratio measurements by ICP-MS
and LA-ICP-MS on biological samples
and single particles. Int J Mass Spectrom
2008; 270: 1-7.
Bosnak C, Pruszkowski E, Neubauer K.
The Analysis of Food Substances by ICPMS. Advanstar Publication, 2008.
Dabrio M, Rodriguez AR, Bordin G et
al. Study of complexing properties of the
α and βmetallothioneins domains with
cadmium and/or zinc using electrospray
ionisation mass spectrometry. Anal Chim
Acta 2001; 435: 319-330.
Leopold I, Gut her D. Investigation of
the binding properties of heavy-metalpeptide complexes in plant cell cultures
CNR Environment and Health Inter-departmental Project
using HPLC-ICP-MS. Fresenius J Anal
Chem 1997; 359: 364-370.
97. McSheehy S, Mester Z. The speciation
of natural tissues by electrospray massspectrometry. II: bioinduced ligands and
environmental contaminants. Trends
Anal Chem 2003; 22: 311-326.
98. Nischwitz V, Michalke B, Kettrup A.
Investigation on extraction procedures for
Pt species from spiked road dust samples
using HPLC-ICP-MS detection. Anal
Chim Acta 2004; 521: 87-98.
99. Rivero Martino FA, Fernandez-Sanchez
ML, Sanz-Medel A. Multi-elemental
fractionation in milk whey by size
exclusion chromatography coupled on
line to ICP-MS. J Anal At Spectrom 2002;
17: 1271-1277.
100. Rottman L, Heumann KG. Determination
of heavy metal interactions with dissolved
organic materials in natural aquatic system
by coupling a high-performance liquid
chromatography system wiyh an inductively
coupled plasma mass spectrometer. Anal
Chem 1994; 66: 3709-3715.
101. Sahan Y, Basoglu F, Gucer S. ICP-MS
analysis of a series of metals (namely:
Mg, Cr, Co, Ni, Fe, Cu, Zn, Sn, Cd and
Pb) in black and green olive samples from
Bursa, Turkey. Food Chem 2007; 105:
102. Shiraishi K. Multi-element analyusis of
18 food groups using semi-quantitative
ICP-MS. J Radioanalytical Nuclear
Chemistry 1998; 238: 67-73.
103. Skelly Frame EM, Uzgris EE. Gadolinium
determination in tissue samples by
inductively coupled plasma mass
spectrometry and inductively coupled
plasma atomic emission spectrometry
in evaluation of the action of magnetic
resonance imaging contrast agent.
Analyst 1998; 123: 675-680.
104. Swami K, Judd CD, Orsini J, Yang KX,
Husain L. Microwave assisted digestion
of atmospheric aerosol samples followed
by inductively coupled plasma mass
spectrometry determination of trace
elements. Fresenius J Anal Chem 2001;
369. 63-70.
105. Urvoas A, Amekraz B, Moulin C, Le
Clainche L, Stocklin R, Moutiez M. Analysis
of the metal-binding selectivity of the
metallochaperone CopZ from Enterococcus
hirae by electrospray ionization mass
spectrometry. Rapid Commun Mass
Spectrom 2003; 17: 1889-1896.
106. Vanhaecke F, Saverwyns S, Wannemacker
G, Moens L, Dams R. Comparision of the
application of higher mass resolution and
cool plasma conditions to avoid spectral
interference in Cr (III)/Cr (IV) speciation
by means of high-performance liquid
mass spectrometry. Anal Chim Acta
2000; 419: 55-64.
107. Wilbur S, Yamanaka M. Simple, Rapid
Analysis of Trace Metals in Foods Using
the Agilent 7700x ICP-MS. Agilent
Technologies Inc, 2009.
108. Wrobel K, Kannamkumarath SS, Wrobel
K, Caruso JA. Hydrolysis of proteins
with methanesulfonic acid for improved
HPLC-ICP-MS determination of selenomethionine in yeast and nuts. Anal
Bioanal Chem 2003; 375: 133-138.
109. Taniguchi S, Shionoya I, Toyama O,
Hayakawa T. Micro-Analysis of Lithium
by Isotope dilution method. Studies on
Mass Spectroscopy 1962; 108-109.
110. Waidmann E, Hilpert K, Stoeppler M.
Thallium determination in reference
materials by Isotope Dilution Mass
Spectrometry (IDMS) using thermal
ionization. Fresenius J Anal Chem 1990;
338: 572-574.
111. Wieser ME, DeLaeter JR. Molybdenum
concentrations measured in eleven
USGS geochemical reference material by
Isotope Dilution Thermal Ionization Mass
Spectrometry. Geostandards Newsletter
2000; 275-279.
112. Yagi M, Masumoto K. Determination
of Strontium in Biological Materials by
Charged Particle Activation Analysis
using the Stable-Isotope Dilution Method.
Cyric Annual Report 1983.
113. Ciceri E, Recchia S, Dossi C, Yang L,
Sturgeon RE. Validation of an isotope
dilution, ICP-MS method based on internal
Monitoring Contaminants in Food Chain and their Impact on Human Health
mass bias correction for the determination
of trace concentrations of Hg in sediment
cores. Talanta 2008; 74: 642-647.
114. Lee SH, Suh JK, Lee SH. Determination
of mercury in tuna fish tissue using isotope
dilution-inductively coupled plasma mass
spectrometry. Microchemical J 2005; 80:
115. Veillon C, Patterson KY, Rubin MA,
Moser-Veillon PB. Determination of
Natural and Isotopically Enriched
Chromium in Urine by Isotope Dilution
Gas Chromatography/Mass Spectrometry.
Anal Chem 1994; 66: 856-860.
116. Al-Harahsheh M, Kingman SW.
Microwave assisted leaching - a review.
Hydrometallurgy 2004; 73: 189-203.
117. Alvarez MB, Malla ME, Batistoni DA.
Comparative assessment of two sequential
chemical extraction schemes for the
fractionation of cadmium, chromium,
lead and zinc in surface coastal sediments.
Fresenius J Anal Chem 2001; 369: 81-90.
118. Greenberg RR, Kingston HM, Watters
RL, Pratt KW. Dissolution problems with
botanical reference materials. Fresenius J
Anal Chem 1990; 338: 394-398.
119. Hullebusch ED, Utomo S, Zandvoort MH,
PN LL. Comparison of three sequential
extraction procedures to describe metal
fractionation in anaerobic granular
sludges. Talanta 2005; 65: 549-558.
120. Peakall D, Burger J. Methodologies for
assessing exposure to metals: speciation,
bioavailability of metals, and ecological
host factors. Ecotoxicol Environ Saf
2003; 56: 110-121.
121. Perez Cid
Fernandez Albores BA,
Fernandez Gomez Falque Lopez E.
Metal fractionation in olive oil and urban
sewage sludges using the three-stage
BCR sequential extraction method and
microwave single extractions. Analyst
2001; 126: 1304-1311.
122. Heumann KG. Isotope-dilution ICPMS for trace element determination and
speciation: from a reference method to
a routine method? Anal Bioanal Chem
2004; 378: 318-329.
123. Watters RLJ, Eberhardt KR, Beary ES,
Fassett JD. Protocol for Isotope Dilution
using inductively coupled plasmamass spectrometry (ICP-MS) for the
determination of inorganic elements.
Metrologia 1997; 34: 87-96.
124. Chu M, Beauchemin D. Simple method to
assess the maximum bio-accessibility of
elements from food using flow injection
and inductively coupled plasma mass
spectrometry. J Anal At Spectrom 2004;
19: 1213-1216.
125. Hansen EH, Wang J. Implementation of
suitable flow injection/sequential injectionsample
schemes for determination of trace
metal concentration using detection
by electrothermal atomic absorption
spectrometry and inductively coupled
plasma mass spectrometry. Anal Chim
Acta 2002; 467: 3-12.
126. Winefordner JD, Gornushkin IB, Correl
T, Gibb E, Smith BW, Omenetto N.
Comparing several atomic spectrometric
methods to the super stars: special
emphasis on laser induced breakdown
spectrometry, LIBS, a future super star. J
Anal At Spectrom 2004; 19: 1061-1083.
127. Jackson BP, Hopkins WA, Baionno
J. Laser ablation-ICP-MS analysis of
dissected tissue: a conservation-minded
approach to assessing contaminant
exposure. Environ Sci Technol 2003; 37:
128. Raith A, Hutton RC. Quantitation
methods using laser ablation ICP-MS.
Part 1: analysis of powders. Fresenius J
Anal Chem 1994; 350: 242-246.
129. Fortunato G, Wunderli S. Evaluation of
the combined measurement uncertainty
in isotope dilution by MC-ICP-MS. Anal
Bioanal Chem 2003; 377: 111-116.
130. Waight T, Baker J, Peate D. Sr isotope
ratio measurements by double-focusing
MC-ICPMS: techniques, observations
and pitfalls. Int J Mass Spectrom 2002;
131. Vandecasteele C, Block CB. Modern
Methods for Trace Element Determination.
Chichester, John Wiley, 1997.
132. Reilly C. Metal contamination of food. Its
CNR Environment and Health Inter-departmental Project
significance for food quality and human
health. Oxford, Blackwell Science, 2002.
133. Swedenborg E, Ruegg J, Makela S,
Pongratz I. Endocrine disruptive
chemicals: mechanisms of action and
involvement in metabolic disorders. J
Mol Endocrinol 2009; 43: 1-10.
134. Yang M, Park MS, Lee HS. Endocrine
disrupting chemicals: human exposure
and health risks. J Environ Sci Health C
Environ Carcinog Ecotoxicol Rev 2006;
24: 183-224.
135. Fromme H, Albrecht M, Boehmer S et
al. Intake and body burden of dioxin-like
compounds in Germany: the INES study.
Chemosphere 2009; 76: 1457-1463.
136. Covaci A, Koppen G, Van Cleuvenbergen
R et al. Persistent organochlorine
pollutants in human serum of 50-65 years
old women in the Flanders Environmental
and Health Study (FLEHS). Part 2:
Correlations among PCBs, PCDD/
PCDFs and the use of predictive markers.
Chemosphere 2002; 48: 827-832.
137. Iida T, Todaka T, Hirakawa H et al.
Concentration and distribution of dioxins
and related compounds in human tissues.
Chemosphere 2007; 67: S263-271.
138. Landi MT, Needham LL, Lucier G,
Mocarelli P, Bertazzi PA, Caporaso N.
Concentrations of dioxin 20 years after
Seveso. Lancet 1997; 349: 1811.
139. Mocarelli P, Needham LL, Marocchi A
et al. Serum concentrations of 2,3,7,8tetrachlorodibenzo-p-dioxin and test results
from selected residents of Seveso, Italy. J
Toxicol Environ Health 1991; 32: 357-366.
140. Van den Berg M, Birnbaum L, Bosveld
A, . Toxic equivalency factors (TEFs) for
PCBs, PCDDs, PCDFs for humans and
wildlife. Environ Health Perspect 1998;
106: 775-792.
141. WHO. Consultation on assessment of healt
risk of dioxins; re-evaluation of the tolerable
daily intake (TDI). Food Additives and
Contaminats 1998; 17: 223-240.
142. Cerna M, Kratenova J, Zejglicova K et
al. Levels of PCDDs, PCDFs, and PCBs
in the blood of the non-occupationally
exposed residents living in the vicinity of
a chemical plant in the Czech Republic.
Chemosphere 2007; 67: S238-246.
143. Nakamura T, Nakai K, Matsumura T,
Suzuki S, Saito Y, Satoh H. Determination
of dioxins and polychlorinated biphenyls
in breast milk, maternal blood and cord
blood from residents of Tohoku, Japan.
Sci Total Environ 2008; 394: 39-51.
144. Santelli F, Boscaino F, Cautela D,
Castaldo D, Malorni A. Determination
of polychlorinated dibenzo-p-dioxins
(PCDDs), polychlorinated dibenzo-pfurans (PCDFs) and polychlorinated
biphenyls (PCBs) in buffalo milk and
mozzarella cheese. Eur Food Res Technol
2006; 223: 51-56.
145. Nelson
CytochromeP450html 2003.
146. Gonzalez FJ, Nebert DW. Evolution of
the P450 gene superfamily: animal-plant
‘warfare’, molecular drive and human
genetic differences in drug oxidation.
Trends Genet 1990; 6: 182-186.
147. Guengerich FP, Shimada T. Oxidation
of toxic and carcinogenic chemicals by
human cytochrome P-450 enzymes.
Chem Res Toxicol 1991; 4: 391-407.
148. Zhou S, Koh HL, Gao Y, Gong ZY, Lee
EJ. Herbal bioactivation: the good, the bad
and the ugly. Life Sci 2004; 74: 935-968.
149. James MO, Sacco JC, Faux LR. Effects
of Food Natural Products on the
Biotransformation of PCBs. Environ
Toxicol Pharmacol 2008; 25: 211-217.
150. Goksoyr A, Forlin L. The cytochrome
P450 system in fish, aquatic toxicology
and environmental monitoring. Aquat
Toxicol 1992; 22: 287-312.
151. Longo V, Marini S, Salvetti A, Angelucci
S, Bucci S, Gervasi PG. Effects of betanaphthoflavone,
dichlobenil on the drug-metabolizing
system of liver and nasal mucosa of
Italian water frogs. Aquat Toxicol 2004;
69: 259-270.
152. Raucy JL, Schultz ED, Wester MR et al.
Human lymphocyte cytochrome P450 2E1,
a putative marker for alcohol-mediated
changes in hepatic chlorzoxazone activity.
Monitoring Contaminants in Food Chain and their Impact on Human Health
Drug Metab Dispos 1997; 25: 1429-1435.
153. Pucci L, Chirulli V, Marini S et al.
Expression and activity of CYP2E1 in
circulating lymphocytes are not altered
in diabetic individuals. Pharmacol Res
2005; 51: 561-565.
154. Danzo BJ. The effects of environmental
hormones on reproduction. Cell Mol Life
Sci 1998; 54: 1249-1264.
155. Alzieu C. Environmental impact of TBT:
the French experience. Sci Total Environ
2000; 258: 99-102.
156. Rhind SM. Endocrine disruptors and
other food-contaminating environmental
pollutants as risk factors in animal
reproduction. Reprod Domest Anim
2008; 43 Suppl 2: 15-22.
157. Soto AM, Sonnenschein C, Chung KL,
Fernandez MF, Olea N, Serrano FO. The
E-SCREEN assay as a tool to identify
estrogens: an update on estrogenic
environmental pollutants. Environ Health
Perspect 1995; 103 Suppl 7: 113-122.
158. Engelman F. Invertebrates: hormoneregulated
Epple A, Scanes CG, Stentson MH
(eds). Perspectives in comparative
Ottawa, Canada,
Academic Press, 1994, pp 36-40.
159. Huberman A. Shrimp endocrinology. A
review. Aquaculture 2000; 191;:191-208.
160. Rempel MA, Schlenk D. Effects
antiandrogens on endocrine function,
gene regulation, and health in fish. Int
Rev Cell Mol Biol 2008; 267: 207-252.
161. Tosti E, Di Cosmo A, Cuomo A, Di
Cristo C, Gragnaniello G. Progesterone
induces activation in Octopus vulgaris
spermatozoa. Mol Reprod Dev 2001; 59:
162. De Loof A, De Clerk A. Vertebrate-type
steroids in Arthropods: Identification,
concentrations and possible functions. In:
Ponchet LM (ed). Advances in Invertebrate
Amsterdam, Elsevier
Science Publications, 1986, pp 117–123.
163. Sandor T, : pp . Steroids in invertebrates.
In: Clark WH, Jr,, Adams TS (eds).
Advances in Invertebrate Reproduction,
Amsterdam, New York, Amsterdam,
North Holland, Inc, 1980, pp 81–96.
164. Voogt PA, Oudejans RCHM, Broertjes JJS.
Steroids and reproduction in starfish. In:
Engels W (ed). Advances in Invertebrate
Amsterdam, Elsevier
Science Publishers, 1984, pp 151-161.
165. Couch EF, Hagino N, Lee JW.
Changes in estradiol and progesterone
immunoreactivity in tissues of the lobster
(Homarus americanus) with developing
and immature ovaries. Comp Biochem
Physiol A 1987; 87: 765–770.
166. Quinitio ET, Yamauchi K, Hara A, Fuji
A. Profiles of progesterone- and estradiollike substances in the hemolymph of
female Pandalus kessleri during an
annual reproductive cycle. Gen Comp
Endocrinol 1991; 81: 343-348.
167. Quinitio ET, Hara A, Yamauchi K, Nakao
S. Changes in the steroid hormone and
vitellogenin levels during the gametogenic
cycle of the giant tiger shrimp, Penaeus
monodon. Comp Bochem Physiol 1994;
109C: 21-26.
168. Yano I, Chinzei Y. Ovary is the site of
vitellogenin synthesis in Kuruma prawn
Penaeus japonicus. Comp Biochem
Physiol 1985; 86B: 213–218.
169. Yano I. Effect of 17b-hydroxyprogesterone on vitellogenin secretion
in kuruma prawn, Penaeus japonicus.
Aquaculture 1987; 61: 49–57.
170. Ghosh D, Ray AK. 17 beta-Hydroxysteroid
dehydrogenase activity of ovary and
hepatopancreas of freshwater prawn,
Macrobrachium rosenbergii: relation to
ovarian condition and estrogen treatment.
Gen Comp Endocrinol 1993; 89: 248-254.
171. D’Aniello A, Di Cosmo A, Di Cristo
C, Assisi L, Botte V, Di Fiore MM.
Occurrence of sex steroid hormones and
their binding proteins in Octopus vulgaris
lam. Biochem Biophys Res Commun
1996; 227: 782-788.
172. Di Cosmo A, Di Cristo C, Paolucci M.
Sex steroid hormone fluctuations and
morphological changes of the reproductive
system of the female of Octopus vulgaris
throughout the annual cycle. J Exp Zool
CNR Environment and Health Inter-departmental Project
2001; 289: 33-47.
173. Di Cosmo A, Di Cristo C, Paolucci
M. A estradiol-17beta receptor in the
reproductive system of the female of
Octopus vulgaris: characterization and
immunolocalization. Mol Reprod Dev
2002; 61: 367-375.
174. Di Cosmo A, Paolucci M, Di Cristo C,
Botte V, Ciarcia G. Progesterone receptor
in the reproductive system of the female
of Octopus vulgaris: characterization and
immunolocalization. Mol Reprod Dev
1998; 50: 451-460.
175. Delsuc F, Brinkmann H, Chourrout
D, Philippe H. Tunicates and not
cephalochordates are the closest living
relatives of vertebrates. Nature 2006; 439:
176. Tosti E, Romano G, Buttino I, Cuomo A,
Ianora A, Miralto A. Bioactive aldehydes
from diatoms block the fertilization
current in ascidian oocytes. Mol Reprod
Dev 2003; 66: 72-80.
177. Cuomo A, Di Cristo C, Paolucci M, Di
Cosmo A, Tosti E. Calcium currents
correlate with oocyte maturation during
the reproductive cycle in Octopus
vulgaris. J Exp Zool A Comp Exp Biol
2005; 303: 193-202.
178. Cuomo A, Silvestre F, De Santis R, Tosti
E. Ca2+ and Na+ current patterns during
oocyte maturation, fertilization, and early
developmental stages of Ciona intestinalis.
Mol Reprod Dev 2006; 73: 501-511.
179. Tosti E, Boni R, Cuomo A. Ca(2+)
current activity decreases during meiotic
progression in bovine oocytes. Am J
Physiol Cell Physiol 2000; 279: C17951800.
180. Hsieh MH, Breyer BN, Eisenberg
ML, Baskin LS. Associations among
hypospadias, cryptorchidism, anogenital
distance, and endocrine disruption. Curr
Urol Rep 2008; 9: 137-142.
181. Scientific Committee on Food. Opinion of
the Scientific Committee on Food on the
Risk Assessment of Dioxins and Dioxinlike PCBs in Food. Brussels, Belgium;
2001. Report No.: CS/CNTM/DIOXIN/20
182. Bilau M, Matthys C, Bellemans M, De
Neve M, Willems JL, De Henauw S.
Reproducibility and relative validity
of a semi-quantitative food frequency
questionnaire designed for assessing
the intake of dioxin-like contaminants.
Environ Res 2008; 108: 327-333.
183. Bocio A, Domingo JL, Falco G, Llobet
JM. Concentrations of PCDD/PCDFs
and PCBs in fish and seafood from the
Catalan (Spain) market: estimated human
intake. Environ Int 2007; 33: 170-175.
184. Charnley G, Doull J. Human exposure
to dioxins from food, 1999-2002. Food
Chem Toxicol 2005; 43: 671-679.
185. Domingo JL, Schuhmacher M, Granero
S, Llobet JM. PCDDs and PCDFs in
food samples from Catalonia, Spain.
An assessment of dietary intake.
Chemosphere 1999; 38: 3517-3528.
186. Fattore E, Fanelli R, Turrini A, di
Domenico A. Current dietary exposure
polychlorodibenzofurans, and dioxin-like
polychlorobiphenyls in Italy. Mol Nutr
Food Res 2006; 50: 915-921.
187. Liem AK, Furst P, Rappe C. Exposure
of populations to dioxins and related
compounds. Food Addit Contam 2000;
17: 241-259.
188. Patandin S, Dagnelie PC, Mulder PG et
al. Dietary exposure to polychlorinated
biphenyls and dioxins from infancy until
adulthood: A comparison between breastfeeding, toddler, and long-term exposure.
Environ Health Perspect 1999; 107: 4551.
189. Schecter A, Cramer P, Boggess K et al.
Intake of dioxins and related compounds
from food in the U.S. population. J Toxicol
Environ Health A 2001; 63: 1-18.
190. Tard A, Gallotti S, Leblanc JC, Volatier
JL. Dioxins, furans and dioxin-like PCBs:
occurrence in food and dietary intake in
France. Food Addit Contam 2007; 24:
191. Weijs PJ, Bakker MI, Korver KR, van Goor
Ghanaviztchi K, van Wijnen JH. Dioxin
and dioxin-like PCB exposure of nonbreastfed Dutch infants. Chemosphere
Monitoring Contaminants in Food Chain and their Impact on Human Health
2006; 64: 1521-1525.
192. Wittsiepe J, Schrey P, Wilhelm M. Dietary
intake of PCDD/F by small children with
different food consumption measured by
the duplicate method. Chemosphere 2001;
43: 881-887.
193. Taioli E, Marabelli R, Scortichini G et al.
Human exposure to dioxins through diet in
Italy. Chemosphere 2005; 61: 1672-1676.
194. European
Regulation 2375/2001 of 29 November
2001 amendingCommission Regulation
466/2001 setting maximum levels for
certain contaminants in foodstuffs.
Official Journal L 321 2001; 1-5.
195. Mayer R. PCDD/F levels in food and
canteen meals from southern Germany.
Chemosphere 2001; 43: 857-860.
196. Kiviranta H, Hallikainen A, Ovaskainen
ML, Kumpulainen J, Vartiainen T.
Dietary intakes of polychlorinated
dibenzo-p-dioxins, dibenzofurans and
polychlorinated biphenyls in Finland.
Food Addit Contam 2001; 18: 945-953.
197. Tsutsumi T, Yanagi T, Nakamura M et al.
Update of daily intake of PCDDs, PCDFs,
and dioxin-like PCBs from food in Japan.
Chemosphere 2001; 45: 1129-1137.
198. Llobet JM, Domingo JL, Bocio A, Casas
C, Teixido A, Muller L. Human exposure
to dioxins through the diet in Catalonia,
Spain: carcinogenic and non-carcinogenic
risk. Chemosphere 2003; 50: 1193-1200.
199. European Commission. Annual EU-wide
Pesticide Residues Monitoring Report 2006. 2008; 1-5.
200. European Commission. COMMISSION
REGULATION (EC) No 1881/2006.
Setting maximum levels for certain
contaminants in foodstuffs. 2006.
201. European Commission. Commission
Regulation (EC) No 333/2007 of 28
March 2007 laying down the methods
of sampling and analysis for the official
control of the levels of lead, cadmium,
mercury, inorganic tin, 3-MCPD and
benzo(a)pyrene in foodstuffs (Text with
EEA relevance ). 2007.
202. Wacholder S, Rothman N, Caporaso N.
Population Stratification in Epidemiologic
Studies of Common Genetic Variants and
Cancer: Quantification of Bias J Natl
Cancer Inst 2000; 92: 1151–1158.
203. Calabrese EJ. Should hormesis be the
default model in risk assessment? Hum
Exp Toxicol 2005; 24: 243.
204. Kaiser J. Hormesis. A healthful dab of
radiation? Science 2003; 302: 378.
205. Waalkes MP. Cadmium carcinogenesis.
Mutat Res 2003; 533: 107-120.
206. Hart BA, Lee CH, Shukla GS et al.
Characterization of cadmium-induced
apoptosis in rat lung epithelial cells:
evidence for the participation of oxidant
stress. Toxicology 1999; 133: 43-58.
207. He X, Chen MG, Ma Q. Activation of
Nrf2 in defense against cadmium-induced
oxidative stress. Chem Res Toxicol 2008;
21: 1375-1383.
208. Liu F, Jan KY. DNA damage in arseniteand cadmium-treated bovine aortic
endothelial cells. Free Radic Biol Med
2000; 28: 55-63.
209. Liu J, Kershaw WC, Klaassen CD. Rat
primary hepatocyte cultures are a good
model for examining metallothioneininduced tolerance to cadmium toxicity. In
Vitro Cell Dev Biol 1990; 26: 75-79.
210. Amara S, Abdelmelek H, Garrel C et al.
Preventive effect of zinc against cadmiuminduced oxidative stress in the rat testis. J
Reprod Dev 2008; 54: 129-134.
211. Kayama F, Yoshida T, Elwell MR, Luster
MI. Role of tumor necrosis factor-alpha in
cadmium-induced hepatotoxicity. Toxicol
Appl Pharmacol 1995; 131: 224-234.
212. Manca D, Ricard AC, Tra HV, Chevalier
G. Relation between lipid peroxidation
and inflammation in the pulmonary
toxicity of cadmium. Arch Toxicol 1994;
68: 364-369.
213. Yamano T, DeCicco LA, Rikans LE.
Attenuation of cadmium-induced liver
injury in senescent male fischer 344 rats:
role of Kupffer cells and inflammatory
cytokines. Toxicol Appl Pharmacol 2000;
162: 68-75.
214. Kamiyama T, Miyakawa H, Li JP et al.
Effects of one-year cadmium exposure
on livers and kidneys and their relation
CNR Environment and Health Inter-departmental Project
to glutathione levels. Res Commun Mol
Pathol Pharmacol 1995; 88: 177-186.
215. Thijssen S, Cuypers A, Maringwa J et
al. Low cadmium exposure triggers a
biphasic oxidative stress response in mice
kidneys. Toxicology 2007; 236: 29-41.
216. Kawanishi S, Hiraku Y, Murata M,
Oikawa S. The role of metals in sitespecific DNA damage with reference
to carcinogenesis. Free Radic Biol Med
2002; 32: 822-832.
217. Shi X, Chiu A, Chen CT, Halliwell B,
Castranova V, Vallyathan V. Reduction
of chromium(VI) and its relationship to
carcinogenesis. J Toxicol Environ Health
B Crit Rev 1999; 2: 87-104.
218. Standeven AM, Wetterhahn KE. Possible
role of glutathione in chromium(VI)
metabolism and toxicity in rats. Pharmacol
Toxicol 1991; 68: 469-476.
219. Bagchi D, Vuchetich PJ, Bagchi M et al.
Induction of oxidative stress by chronic
administration of sodium dichromate
[chromium VI] and cadmium chloride
[cadmium II] to rats. Free Radic Biol Med
1997; 22: 471-478.
220. Bagchi D, Stohs SJ, Downs BW, Bagchi
M, Preuss HG. Cytotoxicity and oxidative
mechanisms of different forms of
chromium. Toxicology 2002; 180; 5-22.
221. Asatiani N, Sapojnikova N, Abuladze M
et al. Effects of Cr(VI) long-term and lowdose action on mammalian antioxidant
enzymes (an in vitro study). J Inorg
Biochem 2004; 98: 490-496.
222. Raghunathan VK, Tettey JN, Ellis EM,
Grant MH. Comparative chronic in
vitro toxicity of hexavalent chromium
to osteoblasts and monocytes. J Biomed
Mater Res A 2009; 88: 543-550.
223. Raghunathan VK, Grant MH, Ellis
EM. Changes in protein expression
associated with chronic in vitro exposure
of hexavalent chromium to osteoblasts
and monocytes: A proteomic approach. J
Biomed Mater Res A 2009.
224. Iavicoli I, Fontana L, Bergamaschi A. The
effects of metals as endocrine disruptors.
J Toxicol Environ Health B Crit Rev 2009;
12: 206-223.
225. Smith VK, Evans MF. Econonic
implications of hormesis: some additional
thoughts. Hum Exp Toxicol 2004; 23:
285-287; discussion 303-285.
226. Calabrese EJ, Baldwin LA. Inorganics
and hormesis. Crit Rev Toxicol 2003; 33;
227. Hammitt JK. Economic implications of
hormesis. Hum Exp Toxicol 2004; 23:
267-278; discussion 279-280, 303-265.
228. Renn O. Hormesis and risk communication.
Hum Exp Toxicol 2003; 22;:3-24.
229. Rodricks JV. Hormesis and toxicological
risk assessment. Toxicol Sci 2003; 71:
230. Zhang Q, Pi J, Woods CG, Andersen ME.
Phase I to II cross-induction of xenobiotic
metabolizing enzymes: a feedforward
control mechanism for potential hormetic
responses. Toxicol Appl Pharmacol 2009;
237: 345-356.
231. Mattson MP. Hormesis and disease
resistance: activation of cellular stress
response pathways. Hum Exp Toxicol
2008; 27: 155-162.
232. Committee on Bioavailability of
Contaminants in Soils and Sediments
of the National Research Council.
Bioavailability of Contaminants in Soils
and Sediments: Processes, Tools, and
Applications. Washington, DC, U.S.A,
The National Academies Press, 2003.
233. Ishida T, Takeda T, Koga T et al.
Attenuation of 2,3,7,8-tetrachlorodibenzop-dioxin toxicity by resveratrol: a
comparative study with different routes
of administration. Biol Pharm Bull 2009;
32: 876-881.
234. Schumann K, Elsenhans B. The impact of
food contaminants on the bioavailability
of trace metals. J Trace Elem Med Biol
2002; 16: 139-144.
235. Sergent T, Ribonnet L, Kolosova A
et al. Molecular and cellular effects
of food contaminants and secondary
plant components and their plausible
interactions at the intestinal level. Food
Chem Toxicol 2008; 46: 813-841.
The pilot study on Endocrine Disruptors
D.G. Mita
CNR, Institute of Genetics and Biophysiscs Adriano Buzzati Traverso (IGB), Naples, Italy
[email protected]
Endocrine disruptors have been described as “exogenous chemical substances or mixtures that alter the
structure or function(s) of the endocrine system and cause adverse effects at the level of the organism, its
progeny, populations, or subpopulations” (EPA, 1998).
Several experimental studies reported that also very low doses of endocrine disruptors can affect the endocrine
system causing diseases and altering the development of mammalian (humans included) and non-mammalian
species. Among the diseases associated with the exposure to endocrine disruptors cancer, cardiovascular risk,
modulation of adrenal, gonad and thyroid functions, and endometriosis are those that mainly catch the public
concern considering their social cost.
This paper describes the research activity planned in the pilot study on Endocrine Disruptors granted by CNR
in the general contest of the Environment and Health Inter-departmental Project (PIAS-CNR).
Over the past 50 years, some chemical
pollutants, such as pesticides, flame
retardants, alkylphenols, polychlorinated
biphenyls, phthalates and metals have
been released into the environment in an
increasing way. Some of these substances,
owing to their ability of interfering with
hormonal activity, are called “Endocrine
Disruptors” (EDs). According to the
definition proposed by the Agency for
Environmental Protection (EPA) of the
United States, the Endocrine Disruptors
are “Exogenous agents that interfere
with synthesis, release, transport,
binding, action or elimination of natural
hormones responsible for the maintenance
of homeostasis and the regulation
of developmental processes and/or
behavioural problems”. The effects of
these compounds on endocrine functions
in animals, and hence in humans, result in
an increase of the incidence of endocrinerelated cancers, increased risk of
cardiovascular diseases, reduced fertility
and in the alteration of development
processes. Endocrine disruptors reach
living organisms through air, soil and water.
The major route of transmission, however,
remains the aquatic environment, where
these substances bioaccumulate through
the food chain. Even at very small doses
EDs perform their harmful activity (1).
The concerns regarding the exposure
to EDs are mainly due to: 1) the adverse
effects observed in some wild animals,
fish and ecosystems; 2) the increase
of some human diseases related to the
endocrine system; and 3) the alterations
of the endocrine functions observed in
laboratory animals after exposure to
some environmental chemical pollutants.
Already in 1996, the USA-EPA identified
CNR Environment and Health Inter-departmental Project
the endocrine disruptors as one of six
priority areas of research.
Human health effects associated with
the presence of environmental endocrine
disruptors have been recognized throughout
foetal development, loss of reproductive
capacity, changes in sexual behaviour, and
onset of cardiovascular diseases (through
obesity) and endometriosis. In addition,
it has been observed an excessive cell
proliferation and carcinogenesis as well
as effects on the neurological and immune
The Endocrine Disruptors are one of
the major topic of the International and
European research on risk assessment in
food and environmental safety. The major
international agencies have proposed to
study the problems associated with the
exposure to Endocrine Disruptors from
different points of view. Just to give an
example, the International Program for
Chemical Safety (IPCS) of the World
Health Organization in 2002 published
the Global Report Assessment on the
endocrine disruptors knowledge (http://
whose main objective was the critical
review of the scientific evidence of the
association between exposure to Endocrine
Disruptors and the damage to human
health or ecosystems.
Moreover, the Organization for Economic
Co-operation and Development (OECD)
has dedicated its attention mainly to
the development and harmonization of
strategies to identify Endocrine Disruptors
and characterize their effects on humans
and ecosystems with the establishment
of the Working Group of Endocrine
Disruptors Testing and Assessment (http://
www.oecd. org/document/62/0,2340,en_2
Europe has not underestimated the
problem of EDS. The first definition of the
problem took place during the European
Workshop on “The Impact of Endocrine
Disruptors on Human Health and Wildlife
(Weybridge 2-4/12/1996). Ten years later
there was a new European Workshop on
the “Impacts of Endocrine Disruptors”
(Helsinki, 8-10/11/2006).
Resources devoted to the research on
Endocrine Disruptors in the last three
European Research Framework Programs
have been more significant. Among
the major projects, we must remember:
INUENDO, ANEMONE, the cluster of
CREDO projects and the CASCADE
network of excellence.
In Italy, some public research institutions,
such as ISPESL and ISS, supported the
research on endocrine disruptors with
funds from the Ministry of Heath. A recent
survey, organized by the Interuniversity
Consortium INBB and the ISS, evidenced
the existence of more than one hundred
research groups actively working in this
field. The CNR addressed this issue and
several initiatives were promoted in joint
action between the Department of Earth
and Environment and the Department
of Medicine. The most important is
the Environment and Health Interdepartmental Project, PIAS-CNR, under
the responsibility of Dr. Fabrizio Bianchi.
The final goal of PIAS is to understand
the links between pollution sources and
their effects on human health, since, as
previously reported, the environment
directly or indirectly affects human health.
The prevention of environmental origin
diseases requires a number of actions
either on attitudes and lifestyles or on
laws and other institutional measures
The pilot study on Endocrine Disruptors
designed to guarantee the safety of the
population exposed to environmental
hazards. This is the goal of the pilot project
on “Endocrine Disruptors”, activated by
CNR within the scope of action of PIAS,
and concisely described in the following
pages. This study represents the logical
conclusion of a series of PIAS initiatives
taken in this scientific area. The project
sees the participation of three research
units belonging to three CNR Institutes,
namely the Institute of Clinical Physiology
(CNR-IFC, Pisa), the Institute of Genetics
and Biophysics (CNR-IGB, Naples), and
the Water Research Institute (CNR-IRSA,
Many research activities, even very good
ones, have been excluded from this project
solely for the scarcity of available funds.
We hope that the results that will be
produced from this project may serve as a
basis and stimulus for future initiatives on
this research field.
The project is divided into three
experimental lines, that at first glance
might seem unrelated, but that in reality
are converging into a single objective:
the study of the epidemiological and
experimental links between some social
diseases and the exposure to endocrine
disruptors. These social diseases are the
cardiovascular risk, which is among the
principal causes of mortality in Italy, and
endometriosis, which affects about 15% of
women worldwide.
To better analyze the epidemiological link
between exposure to endocrine disruptors
and the above mentioned diseases,
the population of a territorial district
recognized as highly polluted: Gela (Sicily,
Italy) has been chosen as to be studied
from an epidemiological point of view.
Gela is sadly known for its pollution since
air, soil and water are polluted by high
concentrations of Endocrine Disruptors
(with estrogenic or androgenic or arylic
activity) and heavy metals. Perhaps this is
why Gela is characterized by a higher level
of malformations and cancer, kidney and
cardiovascular diseases, as well as diseases
of the reproductive tract and thyroid with
respect to the national average. So Gela
is the ideal place to verify the existence
of a direct link between “Environmental
Pollution and Health”. Since food is the
principal mean by which EDs reach man,
and since the population under study lives
in a coastal-marine area, attention will
be paid to determine the concentration
of some EDs in some fish types of
larger consumption by the indigenous
population. Finally, in vivo experiments
of EDS prenatal exposure in mice will be
carried out in order to verify the possible
occurrence of endometriosis.
We will now describe the experimental
approach planned for each of the three
research lines.
3.1 Line 1: Endocrine Disrupters and
cardiovascular risk, occurrence of
endometriosis, modulation of adrenal,
gonad and thyroid functions in Gela
One hundred adult voluntaries of both
sexes and resident in the Gela area will
be recruited with the aim of studying the
possible relationship between the levels of
toxic pollutants in their biological fluids
and the risk or occurrence of cardiovascular
diseases, as well as alterations of thyroid,
gonad and adrenal functions, using
exposure biomarkers and responses to
a specific questionnaire. In particular,
personal, medical history, lifestyle,
environmental and professional exposure,
will be collected together with the weight,
CNR Environment and Health Inter-departmental Project
height, waist circumference and blood
pressure of each recruited person. Blood
analysis will include: blood count, αPTT,
PT, fibrinogen, PCRD, total cholesterol,
HDL, LDL, triglycerides, blood glucose,
uric acid, creatinine, BUN. Hormones
dosage will be carried out testing for LH,
FSH, PRL, P, E2, DHEA-S, total and free
testosterone, 4α androstenediol, Cortisol
ACTH, Δ4-androstenedione, 17α-OH
Progesterone, TSH, FT3, FT4 , aldosterone,
PRA. A 24 hours urine collection is
planned for the determination of creatinine
clearance, electrolytes, and cortisol.
Women recruited into the study for
indirect signs of endometriosis will fill
in an additional questionnaire on their
pregnancy, offspring and ovarian cycle,
and will also undergo blood sample to
determine peripheral blood markers
of endometriosis such as leukocytes,
macrophages, TNF1alfa, CD3, CD 25, IL1,
On the basis of the number of
adults recruited and
the analysed
biological samples, the research can
improve knowledge about the possible
correlation between the exposure to
some environmental pollutants, typical
of industrial and urban areas, and health
outcomes, with particular reference to
cardiovascular diseases, different forms of
cancer and endometriosis.
3.2 Line 2: Endometriosis and Endocrine
Disruptors: an in vivo experimental study
Endometriosis is among the diseases
supposed to be associated with exposure
to EDs. Endometriosis is a recurrent
and benign gynaecological disorder
characterized by the presence of
endometrial tissue outside the uterine
cavity. Endometriosis tissues are found
on the peritoneal surface in the female
pelvis, on the ovaries, on the recto-vaginal
septum, rarely in the pericardium, pleura,
and even in the brain (2). Recent statistics
report a prevalence of 6-10% among Italian
women, but in patients with pain and/or
infertility the prevalence rises to 35-60%
(3). Several epidemiological data link the
occurrence of endometriosis with exposure
to various types of endocrine disruptors
(4). Reproductive effects were found in
monkeys, mice or rats, exposed during
foetal life to polychlorinated biphenyls and
dioxins (5-7). The possible occurrence of
endometriosis in animals exposed during
the foetal period to Bisphenol A (BPA), one
of the most abundant endocrine disruptors
in the environment, is still unknown.
The aim of this research line is to
experimentally verify the onset of
endometriosis in the offspring of mothers
exposed to endocrine disruptors during
the prenatal and perinatal life. As reported
above, this link has been sufficiently studied
in higher animals and in mice or rats, but
only in connection to dioxin or dioxinlike compounds exposure (5-7), never in
connection to BPA. BPA, on the contrary,
has been used to verify the adverse effects
on male fertility. There are many scientific
papers on BPA estrogenic action (8-12). It
has been reported that BPA exposure causes
diseases in the developing foetus. It was
also found that low levels of BPA exposure
during foetal development, for instance,
induces earlier puberty (9) and affects the
prostate size (13). For this research we will
use BALB-C mice exposed to BPA from the
beginning of gestation, during lactation and
in their early stage of life. The dose-response
dependence will be determined together
with: a) the morphological and functional
changes in the uterus; b) the presence of
endometrial tissue outside the uterus in
exposed offspring; c) BPA concentrations
in some target tissues: muscle, brain, liver,
The pilot study on Endocrine Disruptors
The involvement in this research of the
Italian Endometriosis Foundation will
ensure the transfer of the results to the
medical community. The Foundation
will also permit the access to its national
registers in order to allow the comparison
between the epidemiological data of the
national population and those of the Gela
3.3 Line 3: Determination of Endocrine
Disruptors concentration in fish of wide
consumption coming from an area of high
environmental risk
It is well documented that in more heavily
populated areas or industrial places,
contamination by endocrine disruptors
affects not only the system of surface
and profound waters (14-17) and the
surrounding lands, but also the health of
the animals and plants there living (18,19).
It must be remembered that in some animal
species living in these environments
serious diseases and malformations,
attributable to contamination by substances
that affect the hormone system, have been
found (20,21). Many of these substances
are characterized by high lipophlicity and
resistance to degradation. This means that
many endocrine disruptors accumulate
in living organisms and increase their
concentration along the food chain in the
ecosystem (21,22). Fish consumption is
one of the major routes by which endocrine
disruptors reach the humans (23,24).
In order to answer to this concern, the aim
of this research activity is to determine
the concentrations of some pollutants,
known for their ability to interfere with
the endocrine system, in fish of wide
consumption and catch in the Gela Sea.
The content of Bisphenol A, Octylphenol
and Nonylphenol and two ethoxylates of
Nonylphenol ( mono and diethoxylated)
will be determined. The concentration
levels of polychlorinated biphenyls
arsenic, cadmium and mercury will be
also determined.
From the results that will be reached
during this research activity we hope
to obtain indications on: 1)- the link
between exposure to endocrine disruptors
and some social diseases; and 2)- how
the environment and the diet operate
synergetically in promoting some severe
pathologies in wildlife and humans.
We hope also to goad our legislator into
achieving a greater consciousness on the
danger of these invisible killers so that
they can take useful prevention initiatives.
With the contribution of: A. Baldi (Fondazione
Italiana Endometriosi); E. Fommei (Pisa
University and IFC), G. Iervasi, A. Pierini
(IFC); S. Maffei, C.Vassalle (Fondazione “G.
Monasterio” CNR-Regione Toscana, Pisa); C.
Roscioli, S.Valsecchi, L. Viganò, D. Vignati
(IRSA, Brugherio).
Endometriosis, Cardiovascular Risk, Fish’s
Contamination, Bisphenol A.
vom Saal FS & Hughes C. An extensive
new literature concerning low-dose
effects of bisphenol A shows the need for
a new risk assessment. Environmental
Health Perspectives (2005)113:926-933.
Giudice LC & Kao LC. Endometriosis.
The Lancet (2004) 364: 1789-1799.
Baldi A, Campioni M & Signorile PG.
Endometriosis: pathogenesis, diagnosis,
therapy and association with cancer.
Oncology Reports (2008) 19: 843-846.
Anger DL & Foster WG. The link
between environmental toxicant exposure
and endometriosis. (2008) Frontiers
CNR Environment and Health Inter-departmental Project
Bioscience 13:1578-1593.
Newbold RR, Jefferson WN & PadillaBanks E. Long-term adverse effects of
neonatal exposure to bisphenol A on the
murine female reproductive tract (2007)
Reproductive Toxicology 24: 253-258.
Foster WG. Endocrine toxicants including
2,3,7,8-terachlorodiben zo -p - dioxin
(TCDD) and dioxin-like chemicals and
endometriosis: is there a link? (2008)
J Toxicol Environ Health B Crit Rev.
Rier S & Foster WG. Environmental
dioxins and endometriosis (2003) Semin.
Reprod Med. 21:145-54.
Palanza, PL, Howdeshell KL, Parmigiani
S & vom Saal FS. Exposure to a low
dose of bisphenol A during fetal life or
in adulthood alters maternal behavior
inmice (2002). Environmental Health
Perspectives. 110: 415-22.
Markey CM, Luque EH, Muñoz de
Toro M, Sonnenschein C & Soto AM.
In utero exposure to bisphenol A alters
the development and tissue organization
of the mouse mammary gland (2001).
Biology of Reproduction 65: 1215-1223.
Sakaue M, Ohsako S, Ishimura R,
Kurosawa S, Kurohmaru M, Hayashi
Y, Aoki Y, Yonemoto J, & Tohyama C.
Bisphenol-A affects spermatogenesis in
the adult rat even at a low dose (2001) J.
Occup. Health. 43:185-190.
Howdeshell K, Hotchkiss AK, Thayer
KA, Vandenbergh JG & vom Saal FS.
(1999). Exposure to bisphenol A advances
puberty. Nature 401: 762-764.
Takahashi O & Oishi S. Disposition
of orally administered 2,2-Bis(4hydroxyphenyl)propane (Bisphenol A) in
pregnant rats and the placental transfer
to fetuses (2000) Environmental Health
Perspectives 108:931-935.
Ramos JG, Varayoud J, Sonnenschein C,
Soto AM, Muñoz de Toro M & Luque
EH. Prenatal exposure to low doses
of bisphenol A alters the periductal
stroma and glandular cell function in
the rat ventral prostate (2001) Biology of
Reproduction 65: 1271-1277.
14. Furuichi T, Kannan K, Giesy JP &
Masunaga S. Contribution of known
endocrine disrupting substances to the
estrogenic activity in Tama River water
samples from Japan using instrumental
analysis and in vitro reporter gene assay
(2004) Water Research 38: 491–501.
15. Patrolecco L, Capri S, De Angelis S,
Pagnotta R, Polesello S & Valsecchi
S. Partition of nonylphenol and related
compounds among different aquatic
compartments in Tiber river (central Italy)
(2006). Water, Air, and Soil Pollution 172:
16. Salste L, Leskinen P, Virta M, Kronberg
L. Determination of estrogens and
estrogenic activity in wastewater
effluent by chemical analysis and the
bioluminescent yeast assay (2007) The
Science of the Total Environment 378:
17. Viganò L, Benfenati E, van Cauwenberge
A, Eidem JK, Erratico C, Goksøyr A,
Kloas W, Maggioni S, Mandich A &
Urbatzka R. Estrogenicity profile and
estrogenic compounds determined in river
sediments by chemical analysis, ELISA
and yeast assays (2008) Chemosphere 73:
18. Latorre A, Lacorte S & Barcelo D.
Presence of nonylphenol, octyphenol
and bisphenol a in two aquifers close to
agricultural, industrial and urban areas.
(2003) Chromatographia 57: 111–116.
19. Hewitt M & Servos M. An overview of
substances present in Canadian aquatic
environments associated with endocrine
disruption. (2001) Water Quality Research
Journal of Canada 36:191–213.
20. Viganò L, Farkas A, Guzzella L, Roscioli
C & Erratico C. The accumulation levels
of PAHs, PCBs and DDTs are related in
an inverse way to the size of a benthic
amphipod (Echinogammarus stammeri
Karaman) in the River Po (2007) The
Science of the Total Environment 373:
21. Naert C, Van Peteghem C, Kupper
J, Jenni L & Naegeli H. Distribution
of polychlorinated biphenyls and
The pilot study on Endocrine Disruptors
polybrominated diphenyl ethers in
birds of prey from Switzerland (2007)
Chemosphere 68: 977-987.
22. Hinck JE, Blazer VS, Denslow ND,
Echols KR, Gale RW, Wieser C, May
TW, Ellersieck M, Coyle JJ & Tillitt DE.
Chemical contaminants, health indicators,
and reproductive biomarker responses
in fish from rivers in the Southeastern
United States (2008) The Science of the
Total Environment 390: 538-557.
23. Dougherty CP, Holtz SH, Reinert JC,
Panyacosit L, Axelrad DA & Wood TJ.
Dietary exposures to food contaminants
across the United States (2000)
Environmental Research 84:170-185.
24. Turyk ME, Persky VW, Imm P, Knobeloch
L, Chatterton Jr. P & Anderson HA.
Hormone disruption by PBDEs in adult
male sport fish consumers. Environmental
Health Perspectives (2008) 116: 16351641.
CNR Environment and Health Inter-departmental Project
Pilot study for the assessment of health
effects of the chemical composition of
ultrafine and fine particles in Italy
M.C. Facchinia, F. Cibellab, S. Baldaccic, F. Sprovierid
a. CNR, Institute of atmospheric sciences and climate (ISAC), Bologna, Italy
b. CNR, Institute of biomedicine and molecular immunology (IBIM), Palermo, Italy
c. CNR, Institute of Clinical Physiology (IFC) Pisa, Italy
d. CNR, Institute of Atmospheric Pollution Research (IIA) Monterotondo St. (Roma), Italy
[email protected]
Endocrine disruptors have been described as “exogenous chemical substances or mixtures that alter the
structure or function(s) of the endocrine system and cause adverse effects at the level of the organism, its
progeny, populations, or subpopulations” (EPA, 1998).
Several experimental studies reported that also very low doses of endocrine disruptors can affect the endocrine
system causing diseases and altering the development of mammalian (humans included) and non-mammalian
species. Among the diseases associated with the exposure to endocrine disruptors cancer, cardiovascular risk,
modulation of adrenal, gonad and thyroid functions, and endometriosis are those that mainly catch the public
concern considering their social cost.
This paper describes the research activity planned in the pilot project on Endocrine Disruptors granted by
CNR in the general contest of the Environment and Health Inter-departmental Project (PIAS).
A number of epidemiological studies have
shown a correlation between fine particle
concentration and increased mortality or
morbidity. At the same time, due to the
complex chemical composition and varying
size-distributions of PM10 and PM2.5, a clear
explanation of the mechanisms underlying
the toxic effects of atmospheric particulate
matter is still elusive (1).
Inhalation of particulate matter leads to
pulmonary inflammation and reduction in
lung function (2) with secondary systemic
effects or, after translocation from the lung
into the circulation, to direct toxic effects
on cardiovascular function (3) and on the
coagulation pathway thus contributing to
the onset of coronary events (4). Through
the induction of cellular oxidative stress and
proinflammatory pathways (4), particulate
matter augments the development and
progression of atherosclerosis (5). The
main factor of these adverse health
effects seems to be combustion-derived
nanoparticles that incorporate reactive
organic and transition metal components.
An important source of these particles is
new diesel cars with oxidizing converters,
such as modern taxis in North Europe.
Many epidemiological, human clinical,
and animal studies showed that ultrafine
particles (UFPs) penetrate deeply into the
lungs initiating an inflammatory response
leading to respiratory diseases and may
be absorbed directly into the circulating
blood, causing cardiovascular diseases (6).
Recent studies highlighted the importance
of identifying susceptible sub-populations
and mechanisms of involved effects.
Several chronic clinical conditions are
good candidates to define the population
susceptible to UFP acute effects, while
elevated levels of oxidatively altered
biomolecules are important intermediate
endpoints that may be useful markers in
CNR Environment and Health Inter-departmental Project
hazard characterization of particulates
(7). At present, UFPs are not usually
monitored by air quality stations. Thus,
current epidemiological studies have to
rely on PM10 data.
Previous studies have pointed to
chemical species occurring in trace
amounts, having known carcinogenic and
mutagenic effects like PAHs (8,9,10) or
“heavy” metals (11). Others have focused
on the peculiar properties of ultrafine
particles (with a diameter below 0.1 μm)
to penetrate biological membranes (12).
Overall, despite the increasing amount of
data provided by both laboratory and field
studies, the nature of the aerosol particles
fraction inducing health effects is still a
matter of debate. This issue is important,
because the different aerosol constituents
exhibit distinct sources and emission/
formation processes (13,14). Therefore,
linking toxicological and epidemiological
impacts of atmospheric particulate matter
to their chemical composition is a key to
evaluate effective pollution abatement
strategies (15,16).
The existing networks of stations
concentration, usually PM10 or PM2.5 mass,
are not designed to provide chemical
composition and size-distribution data.
In Italy, only at the two EMEP stations of
Ispra (VA) and Montelibretti (RM), the
chemical composition of PM10 and PM2.5
is routinely measured. At the same time,
an increasing series of data on the aerosol
chemical composition and size-distribution
have been provided by short-term
intensive field studies performed in the
frame of national and European research
projects (17). During these experiments,
state-of-the-art instrumentation has been
deployed for aerosol characterization.
For instance, multi-stage impactors were
used to provide size-resolved chemical
composition data, down to the ultrafine
or quasi-ultrafine size range. At the
same time, the chemical analysis of fine
particulate samples has shown that even
in urban areas the water-soluble fraction
of the aerosol contains large amounts of
poorly characterized organic compounds
(WSOC, “water-soluble organic carbon”),
in contrast to the paradigm of many
toxicological studies which attributes the
organic-soluble and water-soluble fractions
of the aerosol to organic and inorganic
compounds, respectively. On the contrary,
recent findings point to WSOC as a major
agent for aerosol toxicity and oxidizing
properties (18,19).
In summary, by
examining the priorities for the evaluation
of upcoming research activities of the
Italian National Research Council (CNR)
to link atmospheric aerosols composition
and properties to their health effects, at
least two specific key issues can already
be addressed and dedicated to a) ultrafine
particles and b) WSOC.
This pilot study will combine the results
of two advanced activities in the field of
atmospheric ultrafine particles composition
and their toxicological properties, carried
out by CNR-ISAC (CNR Institute of
Atmospheric Sciences and Climate) and
CNR-IIA (CNR Institute of Atmospheric
Pollution Research) (WP1, WP2) with two
new advanced health studies carried out
by CNR-IFC (CNR Institute of Clinical
Physiology) and CNR-IBIM (CNR
Institute of Biomedicine and Molecular
Immunology) (WP3, WP4) aimed at
exploring short-term effects due to air
pollutants exposure in subjects with preexistent arrhythmia and lung diseases.
The results of the specific advanced
environmental and health activities will be
evaluated and integrated in WP5 with the
final aim of designing an integrated Italian
Health effects of the chemical composition of ultrafine and fine particles in Italy
research activity for future projects to be
presented in the frameworks of regional
and national projects funded by European
Union Structural Funds (PON, POR) or
EU Research Funds (EC-FP7).
1. To collect and compile the available
chemical composition data of fine and
ultrafine particles in urban and rural
sites in Italy;
2. To test the oxidative potential of
organic compounds in the watersoluble fraction of submicron aerosol;
3. To evaluate the feasibility of performing
epidemiological studies assessing
short-term effects of exposure to air
pollutants in subjects with pre-existent
arrhythmia in Italy;
4. To evaluate the feasibility of
epidemiological studies assessing
short-term effects of exposure to air
pollutants in subjects with pre-existent
lung diseases in Italy;
5. To provide the background knowledge
to design an integrated research project
aimed at assessing the effects of fine
and ultrafine particles on human health
(to be presented within the framework
of PON, POR and EU-FP7).
WP1 Assessment of the chemical
composition of ultrafine particles and its
variability in urban and rural sites in Italy
based on available multi-stage impactor
data and initial measurements using
Aerosol Mass Spectrometers (AMS).
WP2 Evaluation of methodologies to
measure the oxidative potential of the
water-soluble organic fraction (WSOC) of
the aerosol. (CNR-ISAC, CNR-IIA)
WP3 A pilot study to assess short-term
effects of exposure to air pollutants in
subjects with pre-existent arrhythmia.
WP4 A pilot study to assess short-term
effects of exposure to air pollutants in
subjects with pre-existent lung diseases in
Italy. (CNR-IBIM)
WP5 Critical evaluation of the current
proposal results and design of a common
experimental strategy for an integrated
future project on the health effects of
fine and ultrafine particles (CNR-ISAC,
With the contribution of PS2 participants: F.
Bianchi (CNR-IFC), S. Decesari (CNR-ISAC),
G. Viegi (CNR-IBIM), R. Sicari (CNR-IFC).
Keywords: Ultrafine particles (UFPs),
water-soluble organic carbon (WSOC),
cardiopulmonary diseases.
Russell AG, Brunekreef B. A focus on
particulate matter and health. Environ.
Sci. Technol. 2009; (4)3; (4)620 – (4)625.
McCreanor J, Cullinan P, Nieuwenhuijsen
MJ, Stewart-Evans J, Malliarou E, Jarup
L, Harrington R, Svartengren M, Han IK,
Ohman-Strickland P, Chung KF, Zhang J.
Respiratory effects of exposure to diesel
traffic in persons with asthma. N Engl J
Med. 2007; 357:23(4)8-2358.
Andersen ZJ, Wahlin P, RaaschouNielsen O, Ketzel M, Scheike T, Loft
S, 2008. Size distribution and total
number concentration of ultrafine and
accumulation mode particles and hospital
admissions in children and the elderly in
Copenhagen, Denmark. Occup Environ
Rückerl R, Ibald-Mulli A, Koenig W,
Schneider A, Woelke G, Cyrys J, Heinrich
J, Marder V, Frampton M, Wichmann HE,
Peters A. Air pollution and markers of
inflammation and coagulation in patients
with coronary heart disease. Am J Respir
CNR Environment and Health Inter-departmental Project
Crit Care Med. 2006; 15;173:(4)32-(4)
5. Calderón-Garcidueñas L, Solt AC,
Henríquez-Roldán C, Torres-Jardón R,
Nuse B, Herritt L, Villarreal-Calderón R,
Osnaya N, Stone I, García R, Brooks DM,
González-Maciel A, Reynoso-Robles R,
Delgado-Chávez R, Reed W. Long-term
air pollution exposure is associated with
neuroinflammation, an altered innate
immune response, disruption of the
blood-brain barrier, ultrafine particulate
deposition, and accumulation of amyloid
beta-(4)2 and alpha-synuclein in children
and young adults. Toxicol Pathol. 2008;
6. Forastiere F, Stafoggia M, Picciotto S,
Bellander T, D’Ippoliti D, Lanki T, von
Klot S, Nyberg F, Paatero P, Peters A,
Pekkanen J, Sunyer J, Perucci CA. A
case-crossover analysis of out-of-hospital
coronary deaths and air pollution in
Rome, Italy. Am J Respir Crit Care Med.
2005; 172:15(4)9-1555
7. Møller P, Jacobsen NR, Folkmann JK,
Danielsen PH, Mikkelsen L, Hemmingsen
JG, Vesterdal LK, Forchhammer L, Wallin
H, Loft S. Role of oxidative damage in
toxicity of particulates. Free Radic Res.
8. International Agency fo;r Research
on Cancer IARC (1983). Polynuclear
aromatic compounds. Part I. Chemical,
environmental and experimental data.
Monographs on the evaluation of
carcinogenic risk of chemicals to humans,
vol. 32. (Lyon, IARC, 1983).
9. de Raat W.K, J.P. Boers, G.L. Bakker,
F.A. de Meijere, A. Hooijmeier, P.H.M.
Lohman, G.R. Mohn, 199(4). Contribution
of PAH and some their nitrated derivatives
to the mutagenicity of airborne particles
and coal fly ash. The Science of the Total
Environment , 53, 7-28.
10. Binkovà B, Vesely C, Veselà C, Jelinek R,
Sram RJ. Genotoxicity and embryotoxicity
of urban air particulate matter collected
during winter and summer period in two
different districts of the Czech Republic.
Mutation Research, 2009; (4)(4)0, (4)5192
11. Lin CC, Chen SJ, Huang KL, Hwang
WI, Chien G.P, Lin W.Y. Characteristics
of Metals in Nano/Ultrafine/Fine/Coarse
Particles Collected Beside a Heavily
Trafficked Road. Environ. Sci. Technol.
2005; 39 (21), 8113 – 8122.
12. Oberdörster G, Oberdörster E.and
Oberdörster J. Nanotoxicology: an
emerging discipline evolving from
studies of ultrafine particles. Review.
Environmental Health Perspectives.
2005; 113 (7), 823 – 839.
13. Viana M, Kuhlbusch TAJ, Querol X,
Alastuey A, Harrison RM, Hopke PK et
al. Source apportionment of particulate
matter in Europe: A review of methods
and results. Aerosol Science ; 39, 827 –
14. Chow JC, Watson JG, Kuhns H,
Etyemezian V et al. Source profiles for
industrial, mobile, and area sources in
the Big Bend Regional Aerosol Visibility
and Observational study. Chemosphere.
2004; 5(4), 185 – 208.
15. Morozzi G, Mastrandrea V, Trotta
F, Tonti A, Scardazza F, Cenci E.
Chemical characterization and biological
properties of airborne particulate matter.
Aerobiologia. 2005; 8, (4)51-(4)57.
16. Fabiani R, De Bartolomeo A, Rosignoli
P, Morozzi G, Cecinato A, Balducci
characterization of airborne total
suspended particulate and PM10
organic extracts. Polycyclic Aromatic
Compounds. 2008; 28, (4)86-(4)99.
17. Canepari S, Pietrodangelo A, Perrino C,
Astolfi ML, Marzo ML. Enhancement of
source traceability of atmospheric PM by
elemental chemical fractionation. Atmos.
Environ. 2009 (4)3, (4)75(4) – (4)765.
18. Baltensperger U, Dommen J, Alfarra
MR, Duplissy J, Gaeggeler K, Metzger
A, Facchini MC, Decesari S, Finessi
E, Reinnig C, Schott M, Warnke J,
Hoffmann T, Klatzer B, Puxbaum H,
Geiser M, Savi M, Lang D, Kalberer M,
Geiser T. Combined determination of
the chemical composition and of health
Health effects of the chemical composition of ultrafine and fine particles in Italy
effects of secondary organic aerosols: The
POLYSOA project. Journal of Aerosol
Medicine and Pulmonary Drug Delivery.
2008; 21, 1(4)5 – 15(4).
19. Biswas S, Verma V, Schauer J, Cassee
F, Cho A and Sioutas C. Oxidative
potential of semi-volatile and non volatile
particulate matter (PM) from heavy-duty
vehicles retrofitted with emission control
technologies. Environ. Sci. Technol.
2009; (4)3, 3905 – 3912.
CNR Environment and Health Inter-departmental Project