Genotoksikologijos laboratorija

Genotoxicology Laboratory research trend.                                                                                        

The main research trend is assessment of environmental genotoxicity and cytotoxicity effects in aquatic organisms and evaluation genotoxicity risk levels in different European marine ecosystems.

The main objectives:

• Studies of environmental genotoxicity and cytotoxicity effects in various fish and bivalve species inhabiting different marine ecosystems. The genotoxicity (induction of micronuclei, nuclear buds and nucleoplasmic bridges) and cytotoxicity (induction of fragmented, apoptotic, bi-nucleated, 8-shaped, blebed cells) levels were evaluated in 8 fish species from 35 study stations in the North Sea, in 135 stations located in the Baltic Sea and 10 stations in the Atlantic Ocean, in 13 stations of the Scottish coast, also in bivalve mollusks collected from 58 stations located in the Baltic Sea and in Norwegian Sea.  In the southern regions of the Baltic Sea and North Sea, the monitoring of geno-cytotoxicity was performed in 2001-2012. Marine Water Column Monitoring approach was developed through the caging exposures of fish and mussels in the Norwegian oil fields in the North Sea, also caging of mussels in the Bornholm zone of dumped chemical munitions and in contaminated sites of the Gulf of Bothnia in the Baltic Sea.
• Experimental assessment of geno-cytotoxicity effects caused by different chemical compounds in aquatic organisms. Development of environmental genotoxicity approach through laboratory exposures was performed for the validation micronuclei and other nuclear abnormalities assays as monitoring tools. Experimental exposures applied in halibut, long rough dab and turbot treatments by using different exposure time and concentrations of Norwegian and arctic crude oil and in mussel exposures to crude oil, different heavy metals, PAHs and their mixtures.
• Analysis of contaminants biodegradation by microorganism communities in digestive tract of different fish and mollusk species. Functional and biochemical activity of bacteria was determined in marine and freshwater fish, bivalve and crustacean digestive tract and identification of PAH-degradation bacteria using DNA sequencing methods was carried out.
• Genotoxicity risk assessment in different regions of studied marine ecosystems. There were established the reference and background genotoxicity levels and using the GIS format of the background responses, the scales of low, increased, moderate, high and extremely high responses in different fish and bivalve species was elaborated. Genotoxicity risk assessment was done in fish from different regions of the Baltic Sea, North Sea and Atlantic Ocean. As an outcome, the largest data base on marine genotoxicity response and risk levels was created; time series data in the GIS format are available for studied fish and bivalve species. Special attention was committing to the genotoxicity risk assessment in the chemical munitions dumpsites.
• Development of genotoxicity assays for the monitoring of contaminant effects in marine ecosystems and elaboration indicators for integrated monitoring of contamination and biological effects regarding implementation of EU Marine Strategy Framework Directive in HELCOM and OSPAR zones. Using accumulated database on marine genotoxicity studies in our laboratory, the background documents on micronucleus (MN) assay as a tool for assessing cytogenetic/DNA damage in marine organisms was fulfilled for the HELCOM (Baltic Sea) and OSPAR (North Sea and Mediterranean Sea) areas. Wide-scale and long-term studies of environmental genotoxicity in our laboratory significantly stimulated the elaboration of MN assay and including the approach into the list of core indicators implementing Marine Strategy Framework Directive across the EU countries.

Laboratory is leading the long-time research programme 1.2.5. Investigations of hazardous substances distribution in the ecosystems, regularities and mechanisms of ecotoxic and biological effects under the impact of anthropogenic stress.

Outline of laboratory achievements                                                                     

At first time there were described the regularities of geno-cytotoxicity formation in different native fish and molluscs species and their various tissues, evaluated relationships between geno-cytototoxicity responses and pollution by hazardous substances, biological or environmental variables in studies marine ecosystems. At first time the data on environmental genotoxicity in studied marine ecosystems were presented in GIS format and created 4- and 5-grade scales for the genotoxicity risk assessment in marine ecosystems. Increased genotoxicity risk identified during the Nord Stream pipeline and cable St. Petersburg-Kaliningrad construction, implementation of large mine clearance programme in the Baltic Sea, as well as in zones of the chemical munitions dumpsites, extensive navigation, oil and gas extraction fields, nuclear power plants, estuaries of the large European rivers. There was clarified geno-cytotoxicity formation under the influence of increased pollution and in recovery process.

In multiple experimental exposures (performed mainly in IRIS, Norway), the micronucleus test and induction of the other nuclear abnormalities were validated being sensitive biomarkers,  proved the utility of used tools and provided evidences on genetic risk of various compounds, particularly PAHs, mixtures of alkylphenols and produced water discharged from the offshore oil platforms.

Genotoxicology Laboratory of the Nature Research Centre is owner of the largest scientific database in Europe on marine genotoxicity studies, which was used in creating the background ICES OSPAR and HELCOM documents for the development an integrated approach to the monitoring of chemical contaminants and biological effects and assessment criteria for Descriptor 8 of Good Environmental Status under EU Marine Strategy Framework Directive. The scientific database was used in the EU Parliament in discussion of the Nord Stream project implementation and monitoring of environmental impacts, also Regulation of the European Parliament and of the Council “On safety of offshore oil and gas prospection, exploration and production activities”, Brussels, 2011/0309/COM. The outcomes of our studies were presented during an official state visit of the Lithuanian President to the Finland and were applied in development second stage of the EU BONUS programme.  

International collaboration, traineeships etc.                                               

The laboratory collaborative efforts cover scientific institutions practically from the all nine countries around the Baltic Sea and also include Norway, United Kingdom and France. The wide-scale environmental genotoxicity studies in the North Sea and Atlantic (over 60 study sites) and multiple laboratory validation studies were carried out at IRIS (Norway) experimental facilities together with scientific partners from the other European countries. In 2008-2012 there are published 12 joint publications with co-authors from Norway, Sweden, Finland, Iceland, Estonia, Germany, Italy in international pre-reviewed journals with impact factor and the other publications with co-authors from Denmark, Latvia, Russia.

Traineeships – during the implementation of the BONUS BEAST project we have fulfilled the standard operations for the analysis of micronuclei in marine fish and bivalve molluscs. Following the SOPs, performance is related to dr. Halldora Skarphedinsdottir, Stockholm University and to Jennifer Ipse, vTI Institute of Fisheries Ecology, Germany. On the other hand, our scientists were trained during the Walther Herwig sampling surveys on histopathology and external fish disease biomarker studies. Consultancy on micronuclei test application in integrated monitoring and GES evaluation for the implementing EU WSFD was done for scientists from the other countries.

Consultancy of EU and Lithuanian Parliament members, Ministry of Foreign Affairs of Lithuania.

The outcomes of our studies were useful for the International Scientific Advisory Board (The Hague) activities in preparation and implementation of the UN General Assembly Resolution on Cooperative Measures to Assess and Increase Awareness of Environmental Effects Related to Waste Originating from Sea-Dumped Chemical Munitions A/RES/65/149.