Development of an ELISA for Vitellogenin to assess estrogen exposure in the subantarctic fish Patagonotothen tessellata (Perciformes, Nototheniidae).

FERREIRA, M.F.; GENOVESE, G; FERNANDEZ D; LO NOSTRO, FABIANA; KATSIADAKI, IVANA

Montpellier

Congreso; 14th International Congress on the Biology of Fish (ICBF); 2020

ICBF

The presence of many persistent organic pollutants has been reported in water, sediments and in vertebrates from Antarctic and sub-Antarctic areas;however, information about their potential impact on fish physiology is still scarce. The southernmost city of the world, Ushuaia (Tierra del Fuego,Argentina) and its bay are not the exception to the decline of environmental quality. The black southern cod, Patagonotothen tessellata, belongs to theprevailing suborder of the coastal ichthyofauna (Notothenioidei). In the present work we purified vitellogenin (VTG), a well stablished biomarker ofestrogenic exposure, and develop an immunoassay (ELISA) in order to quantify this protein in plasma and mucus samples. Males sampled in Tierra delFuego National Park received intraperitoneal injections (10 ug/g) with 17b-estradiol every 3 days during 21 days. Size exclusion chromatography wasperformed using a Superdex 200 HR cromatograph column followed by an ion exchange column. Purity of the fraction was determined analyzed bysodium duodecyl sulfate-polyacrylamide gel electrophoresis and the purified VTG fraction was used for rabbit immunization and ELISA standards. Nontreatedmales, estrogenized males as well as reproductively active females were used to validate the method, which showed a high degree of sensitivityand specificity of the assay. This method represents a new tool to evidence estrogenic exposure in Beagle Channel fish.