Assessment of in situ estrogenic effects in the Black chinned Tilapia (Sarotherodon melanotheron) using vitellogenin measurement by ELISA method

Abstract : The aim of this study was to develop an enzyme linked immunosorbent assay (ELISA) to quantify vitellogenin in the Black chinned Tilapia (Sarotherodon melanotheron). Tilapia vitellogenin T-vtg) was first purified from plasma samples of estradiol-induced fish using a two-step chromatographic method. The T-vtg was characterized with an estimated molecular weight of 482 kDa under native conditions. Polyclonal antibodies against T-vtg were raised in rabbits and the specificity of the antibodies to recognized T-vtg was verified by Western blotting and indirect ELISA experiments on blood samples. By using these antibodies and purified T-vtg, a competitive ELISA was set-up and validated. This assay appears sensitive, with a detection limit of 1.81 ng/ml, and specific, as shown by the competition curves, obtained by serially diluting blood of females, that were parallel to the T-vtg standard curves. The ability of the T-vtg ELISA to quantify vtg induction was achieved by in situ experiment. For this purpose, adults males and females tilapia were collected in 8 West African sites and circulating vtg concentration was measured. The present study shows that the developed T-vtg ELISA is a useful tool to assess the effect of estrogenic contamination in wild Black chinned Tilapia.
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Conference papers
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Submitted on : Wednesday, April 2, 2014 - 3:52:44 PM
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  • HAL Id : ineris-00970787, version 1
  • INERIS : EN-2011-113

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Awa Ndiaye, Olivier Palluel, Jean Durand, François Brion, Jean-Marc Porcher, et al.. Assessment of in situ estrogenic effects in the Black chinned Tilapia (Sarotherodon melanotheron) using vitellogenin measurement by ELISA method. 21. SETAC Europe annual meeting "Ecosystem protection in a sustainable world : a challenge for science and regulation", May 2011, Milan, Italy. ⟨ineris-00970787⟩

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