Section 1: PROJECT IDENTIFICATION
Information to be provided for project identification

NOT CONFIDENTIAL

Title of the project

ENVIRONMENTAL AGENTS SUSCEPTIBILITY ASSESSMENT UTILISING EXISTING AND NOVEL BIOMARKERS AS RAPID NON-INVASIVE TESTING METHODS

Acronym of the project: EASYRING

Type of contract: RTD

Total project cost: (in euro)

2351,056€

Contract number:

Duration: (in months)

EU contribution: (in euro)

QLK4-2002-02286

36 Months

1890,209€

Commencement date:

01/01/2003

Period covered by the p. report:

1 January 2003 – 31 December 2004

PROJECT COORDINATOR

Name:

Alberta Mandich

Title:

Dr

Address: V.le Benedetto XV, 5 16132 Genoa, Italy

Telephone:

390103538046

Telefax:

+390103538047

E-mail address:

mandich@unige.it

Key words: (5 maximum - Please include specific keywords that best describe the project.).

Endocrine disrupters, Biomarkers, Risk assessment, in vitro, dipstick

World wide web address: (the project’s www address )

www.easyring.org

List of participants
Provide all partners’ details including their legal status in the contract i.e.,contractor, assistant contractor (to which contractor?).

P1 and Coordinator Department of Environmental Experimental and Applied Biology (DI.BI.S.A.A.)
Team leader: Dr. Alberta MANDICH
Universityof Genoa, Viale Benedetto XV, 5 - 16132 Genoa - Italy
mandich@unige.it

P2 Department of Molecular Biology (UiB)
Team leader: Dr Bjørn-Einar GRØSVIK
University of Bergen, P.O. Box 7800, NO-5020Bergen - Norway
bjorn-einar.grosvik@mbi.uib.no

P3 Biosense Laboratories AS (BIOSENSo)
Team leader: Prof. Anders GOKSØYR
Thormøhlensgate, 55 NO-5008 Bergen - Norway
Anders@biosense.no

P4 Department of Inland Fisheries (FWB.IGB)
Team leader: Prof. Werner KLOAS
Leibniz - Institute of Freshwater, Ecology and Inland Fisheries, Müggelseedamm 310, 12587 Berlin -Germany
werner.kloas@igb-berlin.de

P5 Water Research Institute National Council of Research(IRSA-CNR)
Team leader: Dr. Luigi VIGANO’
Via della Mornera, 25, 20047 Brugherio (Milan) - Italy
vigano@.irsa.rm.cnr.it
Subcontractor:
Dr. Gaetano Gentili
GRAIA Via Repubblica, 2, 21020 Varano Borghi (Varese) - Italy
graia@libero.it
Status: Private Institution

P6 Université de Mons-Hainaut (UMH)
Team leader: Prof. Henri ALEXANDRE
Faculté de Médicine et de Pharmacie, Service de Biologie et Embryologie, Avenue du Champ de Mars, 7
7000 Mons - Belgium
henri.alexandre@umh.ac.be
Subcontractor:
Dr. Anne Van Cauwenberge
Institut Provincial d’Hygiène et
de Bactériologie
Boulevard Sainctelette, 55
B-7000 Mons - Belgium
anne.vancauwenberge@hainaut.be
Status: Public (governmental) Institution

P7 Liverpool John Moores University (JMU)
Team leader: Dr. Mark CRONIN
School of Pharmacy and Chemistry, Byrom Street, Liverpool – L33AF, United Kingdom
M.T.Cronin@livjm.ac.uk

P8 Mario Negri Institute (NEGRI)
Team leader: Dr. Emilio BENFENATI
Laboratory of Environmental Chemistry and Toxicology, Via Eritrea, 62, 20157 Milan - Italy
benfenati@marionegri.it
Subcontractor:
Dr. Giuseppina Gini
Dipartimento di Elettronica e Informazione Politecnico di Milano, Via Ponzio, 34/5, 20133 Milan - Italy
gini@elet.polimi.it
Status: Public (governmental) Institution

Section 2: Project Progress Report
(2 pages maximum.. Use short sentences. Be factual. Avoid technical terms as much as possible)

NOT CONFIDENTIAL

Objectives:

The major objectives and achievements expected for EASYRING are:

• Rapid, non-invasive test development for the identification of specific biomarkers for endocrine disrupting chemicals in the mucus of aquatic species.
• Novel functional biomarker(s) for the identification of endocrine disrupting chemicals.
• Production and validation of new enzyme linked immunosorbent assays (ELISAs) for the detection of new biomarkers in plasma, mucus and tissues of aquatic species.
• Improvement of in vitro test for the screening of endocrine disrupting chemicals.
• Development of analytical methods in GC-MS and LC-MS for selected endocrine disrupting chemicals in environmental water and biota.
• Short/long term and low level exposure effects of selected chemicals and mixture in low vertebrates.
• Short/long term and low level exposure effects of selected chemicals and mixture, through food and water chain, in small mammals.
• QSARs development for the prediction of chemicals able to elicit endocrine disruption.
• QAARs development to extrapolate the responses of the different experimental models.

Results and Milestones:

• Establishment of retinol binding protein and transthyretin as new biomarker in Xenopus (M1)
• Identification of established biomarker proteins (Vtg/) in mucus of exposed fish(M8)
• Partial cloning of DMRT1-like gene of carp and related real time-PCR analytic protocol (M13).
• Development of assays on pre-implantation embryos (M21).
• Collection of data for table of anomalies in early development stages of studied species, caused by long-term exposures to selected chemicals and their mixtures
• Collection of data for table of morphological alterations caused by selected chemicals and mixtures after short and long-term exposures
• Collection of data for table of biochemical alterations caused by selected chemicals and mixtures after short and long-term exposures Table of biochemical alterations caused by selected chemicals and mixtures after short and long-term exposures
• The development of models (QSARs) for the prediction of the endocrine disruption capability of compounds. Comparison of different methodologies for the prediction of activity (M27)
• Website

Benefits and Beneficiaries:

• Improvement of the standard validated tests for risk assessment (beneficiaries: policy makers, scientist, national and regional environmental agencies, environmental quality monitoring agencies and laboratories)
• Development of new non invasive test methods (beneficiaries: policy makers, scientist, national and regional environmental agencies, environmental quality monitoring agencies and laboratories)
• Improvement of in vitro test systems with alternative screening and testing protocols (beneficiaries: scientist, environmental quality monitoring agencies and laboratories)
• Improvement of chemical analytical method and their application (beneficiaries: scientist, environmental quality monitoring agencies and laboratories)
• Improvement of knowledge on Cytotoxic and teratogenic effects of endocrine disrupters/modulators and quantification of exposures in mammals (cause-effects relationships investigation) (beneficiaries: policy makers, national and regional environmental and health agencies, scientist).

Future Actions (if applicable):

For the future, activities will be performed following the original timetable foreseen in the technical annex. Even though, due to the problems arisen during the first year of work especially related to biomarker genesis, the Consortium set up strategies in order to solve the problem and close the gap.

As requested by the referees the Consortium has been set up a dissemination plan. The choice will be to prefer interdisciplinary article produced among partners, in order to obtain a wider view point. So these “macro argument” should be more comprehensive at different scientific approaches. Scientific groups will be engaged to increase the dissemination of the emergent data within the end of 2005.

The third year activities in field will include specific studies on the target species on the two sampling points, located upstream and downstream the LambroRiver confluence. In particular, the activities carrying on are aimed to: study the diet composition of the target species through the analysis of the stomach contents, determine the linear growth rate through the scalimetric method that allows the age determination and determine the absolute and relative fecundity through the count of eggs present in ovary sub-samples. The prototype of dipstick for the detection of Vtg in the mucus of fish will be checked on-site in the field studies planned during these surveys in Spring 2005.

Regarding modeling starting by month 30 the database will be formally closed. At this time will be contain quantitative and qualitative data from various endpoints. In addition it will include a selection of physico-chemical properties and descriptors that have been found to be important in the modelling process. The database will be placed on the Project internet page. The possibility of converting the database using an internet mark-up language (XML) in collaboration with the UK EPSRC project “Pythia” (University of Bradford) will be explored.

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