WP.4 BIOMONITORING IN THE NORTH ATLANTIC SEA
(Dr.Odd-Ketil Andersen)
General objectives:
In the North Atlantic region, some of the core validated biomarkers have been used in environmental monitoring in some cases, but there is large differences in practice between the countries in the region, and there is a need to validate and inter-calibrate more biomarkers to build the necessary suites of biomarkers to be used in both exposure and effect screening exercises.
In this work-package we will focus on development of multiple biomarker suites that can be applied on multiple species with different life strategies in an ecosystem to detect chemical stressors that might affect the function of the ecosystem. It is important that these suites can be applied in different biota with different environmental toxins.
This project will use multiple biomarkers to detect the effects of exposure to various known contaminants present in gradients between a point source and background levels. The sensitivity of detection will be assessed at defined study sites, using suitable key species and different life strategies. This will be performed both at sites where a single type of contaminant dominates in the recipient, and at sites with several contaminants causing a combined effect response in the organisms.
Sites:
The species to be studied are selected because they are widely distributed, economically important, easily sampled and already benefit from knowledge of biomarker responses. In addition it will be important to identify new sensitive species and species with different life strategies in the ecosystem that can be monitored, in which case other species than those listed below will be tested.
The following species will be studied at every site: cod (Gadus morhua), wrasse (Symphodus melops), shore crab (Carcinus maenas), and common mussel (Mytilus edulis). The following species will be studied when present or on selected sites: eelpout (Zoarces viviparus), edible crab (Canser paragus), baltic clam (Macoma balthica).
Methods:
A wide range of biomarkers will be applied and several new ones tested, also biomarkers from the research workpackage WP1. The ones listed will be performed on the relevant species and sex in this study, with the aim to come up with robust suites of biomarkers for the different contamination types. The biomarker responses will be compared against traditional methods as sex, morphometric indices, histopathology, contaminant concentration/bodyburden analyses, benthic community structure, basic hydrography (temperature, salinity, oxygen).
The following list contains both validated core biomarkers and biomarkers to be tested against validated biomarker responses. In addition to this, new biomarkers will be developed and tested throughout the program. Acetylcholinesterase (AChE), CYP 1A levels and EROD activity, PAH bile metabolites, Glutathione metabolism, Oxidative stress enzymes, Multidrug resistance (MDR), Metallothionein induction.
Immunocompetence: macrophage activity and cytotoxic test, Apoptosis, Neutral lipid accumulation, Hydrophobic/aromatic DNA adducts, Micronuclei (MN).
Lysosomal stability:
histochemical approach and Neutral Red assay, Hearth rate monitoring.
Endocrine disorders:
aromatase, vitellogenin (Vtg-yok protein), ZRP (Zona Radiata Protein), hormone shifts and embryo sex ratio, Early life stage tests.
TASK 1: Intercalibration exercises on core biomarkers
Objectives:
- Exchange knowledge between the various partners to discuss common protocols and standardization of methods.
- Methods should be discussed under the responsability of one or several scientific leaders who are considered as the specialists in the field of each biomarker.
- Close relation with the BEQUALM project whose purpose is to develop control procedures and quality assurance for marine biological effects measurements.
- Intercalibration exercise will be carried out using validated and standardized procedures on :Acetylcholinesterase inhibition (AChE), Lysosomal activity, EROD activity, metallothionein content, PAH bile metabolites.
- Same samples will be shared by several laboratories working on the same biomarkers in order to facilitate intercalibration excercises including laboratories involved in the two other biomonitoring Programes (WP2-WP4).
Methods:
The organization of a Workshop will allow different analytical methods and techniques that concern core biomarkers to be compared and standardized under the scientific responsability of a leader per biomarker.
A part of the workshop will be devoted to the specific requirements of sampling in terms of minimum number of samples, conditions of storage etc…
TASK 2: Site survey : preparation, collection of samples and background information
Objectives:
The site responsible partner shall be responsible for the logistics of the field survey and the catching of the necessary biological material in a shape suited for the biomarker analyses to be performed. The preparation includes necessary laboratory accommodations for sampling and on site measurements, and preparation of samples for storage and transport to the respective laboratories. It is assumed that most sites will have three sampling stations in a gradient from the source of pollution and one reference station.
Not all the biomarkers will be sampled both winter and autumn. The winter sampling is mainly for the early life stage tests.
TASK 3: Measurements of biomarkers (core and research) on the target species
Objectives:
Testing of multiple biomarkers on multiple species with different life strategies in the ecosystem at different sites. Same samples will be shared by several laboratories working on the same biomarkers in order to facilitate intercalibration exercises including laboratories involved in the two other biomonitoring Programes (WP2-WP4).
Methods :
Different biomarkers a) core (established) and b) additional (those which needs to be validated in the field) will be studied in primary target species on every site while those in secondary target species will be studied only in selected areas. Each partner is responsible for delivering results and evaluation of the results accord to table WP4. The results will be presented in a format for common data banking.
TASK 4: Measurement of supporting parameters : chemical contaminants in fish and mussel, physiological parameters
Objectives:
- Assessment of the biological effects of contamination on biota in selected Baltic sites of suspected contamination.
- Collection of morphometric indices and physical parameters during sampling (TASK 2)
Methodology:
Specimens of the target species will be collected for tissue contaminant analyses, although only during two sampling occasions. The following common contaminant groups will be addressed: (1) PAH compounds, (2) pesticides (DDT and derivatives), (3) PCB congeners, and (4) heavy metals.
Basically, only existing data on sediment and water contamination will be used for description of the degree and type of contamination at each study site.
TASK 5: Data Handling
Objectives:
- Adjust formats of data obtained from different participants to uniform standard. Distribution of database among potential users. Data storage for future use and documentation.
- Collect data on biomarkers and chemicals obtained during the site experiments (WP2, WP3 and WP4) and to transfer these data to the data base operated by P2.
- Apply common procedures developed by WP5 to achieve a synthetic and quantitative assessment of the health status of marine organisms.
- Refine and apply graphical methods to facilitate between-site and between-survey comparisons.
Methods:
Results are sent to partner 25 as disks and are converted to uniform format. Data are compiled to a database in MS Access format, and distributed as ready for use disks.
The common procedures proposed by WP5 to provide synthetic indexes will be applied to WP2 data and the sensitiveness and robustness of the chosen indices assessed with regard to the Baltic Sea specificity. In situ sources of variation of biomarker responses will be assessed in order to validate sampling strategies in the field. Results of the first surveys can then be used to optimize strategies under the known constraints.
Constraints on sampling effort allocation, s.a. cruise time vs. within-site number of sampling stations, could forbid the use of multivariate analysis on a regional basis. Other ways of summarizing ecotoxicological information and of confronting this information to chemical measurements will be investigated. In particular the Integrated Biomarker Response (IBR), based on star plot area computations, will continue to be developed and refined.
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