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Evaluation of biochemical responses in Palearctic and Lake Baikal endemic amphipod species exposed to toxicity stress

REG-5 Mesoscale models of hydrothermal processes and anthropogenic admixture transferring in atmosphere and hydrosphere. Evaluation of biochemical responses in Palearctic and Lake Baikal endemic amphipod species exposed to toxicity stress. Marina Protopopova , Darya Bedulina

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Evaluation of biochemical responses in Palearctic and Lake Baikal endemic amphipod species exposed to toxicity stress

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  1. REG-5 Mesoscale models of hydrothermal processes and anthropogenic admixture transferring in atmosphere and hydrosphere Evaluation of biochemical responses in Palearctic and Lake Baikal endemic amphipod species exposed to toxicity stress Marina Protopopova, Darya Bedulina Irkutsk State University, Irkutsk Laboratory of biological systems adaptation Bolshie Koty, Baikal, Russia June 25-29, 2007

  2. The evaluation of antioxidant enzymes and sHSP as possible biomarkers for using in the toxicological studies of Baikal endemic species The aim of research

  3. The objects of research Freshwater species of amphipods (Crustacea, Amphipoda): Baikal species E. cyaneus (Dyb.) E. verrucosus (Gerstf) Palearctic species Gammarus lacustris (Sars) Gammarus lacustrisSars

  4. The biomarkers of stress Antioxidant enzymes: • peroxidase (POD) • catalase (CAT) Heat-shock proteins (HSPs): • small Heat-shock proteins (sHSP) Toxic stress solution of cadmium chloride (CdCl2)

  5. Methods Measurement of enzyme activities Spectrophotometricmethod POD activity was measured with guaiacol as the substrate at 436 nm, pH 5 according to Bergmeyer (1983) CAT activity was measured with hydrogen peroxide as the substrate at 240 nm and a pH of 7 according to Aebi (1984).

  6. Methods Measurement of sHSPs content SDS- electrophoresis Western-blotting

  7. Results PODactivities in the three amphipod species exposed to 0,05 mgL-1CdCl2

  8. Results CAT activities in the three amphipod species exposed to 0,05 mgL-1CdCl2

  9. Results Western blot analysis on sHSPs of the amphypods after 24 hours exposition in different CdCl2 solutions line 1 – control, line 2 - 0,05 mgL-1, line 3- 0,5 mgL-1, line 4 – 5 mgL-1. • E. cyaneus, • E. verrucosus, • C.G. lacustris

  10. The chaperon function ofsHSPs stress The refolded protein chain The native protein chain The denaturated protein chain

  11. Conclusion • CdCl2 causes significant effects of the peroxidase. • CdCl2 causes induction of the dose-dependent synthesis of small heat shock proteins. • All measured parameters varied interspecifically and depended on the ability of the species to resist cadmium chloride toxicity. The synthesis of small heat shock proteins and peroxidase activity, could be successfully used as biomarkers for the further studies in amphipod species including endemics from Lake Baikal.

  12. The publication:Evaluation of biochemical responses in Palearctic and Lake Baikal' endemic amphipod species exposed to CdCl2 // Ecotoxicology and Environmental Safety, 2007 (in press)The grants of Russian Foundation for Basic Research:№ 06-04-48099-а, 5-04-97239-baikal, 05-04-97263-baikal

  13. Thank you

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