Assessment of environmental endocrine disruption with plasma concentration of vitellogenin in Koi carp of rivers and wetland sites.

卵黄蛋白原(VTG)是一种雌性特有的血浆蛋白，是卵生脊椎动物卵黄蛋白的主要前体。该蛋白质可以作为环境内分泌干扰的有用生物标记物。为评价河流湿地锦鲤(Cyprinus Carpio)血浆VTG浓度对环境内分泌的干扰程度，将鲤鱼VTG纯化为匀浆，提纯多克隆抗血清，建立了测定血浆VTG浓度的单次放射免疫扩散法和相应的酶联免疫吸附试验(EL ISA)。在Ojiya市Shinano河捕获的24只雄鱼中有2只和在新泻市Shinano河捕获的15只雄鱼中有5只血浆Vtg浓度略高(&gt；1μg/ml)。暴露于污水处理厂污水10-14天的幼公鱼血浆Vtg水平高于未暴露的雄鱼，表明流出物中含有生物活性的雌激素化合物。GC/MS分析表明，出水中的雌二醇-17-β是提高锦鲤血浆VTG水平的主要化合物之一。

Vitellogenin (Vtg) is a female specific plasma protein and the major precursor of egg yolk protein in oviparous vertebrates. The protein can serve as a useful biomarker for environmental endocrine disruption. The present study was carried out to assess environmental endocrine disruption with plasma Vtg concentration in Koi carp (Cyprinus carpio) of rivers and wetland sites.We purified Koi carp Vtg to homogeneity, raised specific polyclonal antiserum to the purified Koi carp Vtg and established a single radial immunodiffusion assay and a homologous enzyme-linked immunosorbent assay (ELISA) to measure plasma Vtg concentration.To assess environmental endocrine disruption, wild Koi carp were collected in the wetland site Sakata and Toyanogata and the Shinano River. Two of 24 males captured in the Shinano River in the Ojiya City area and 5 of 15 males captured in the Shinano River in Niigata City area possessed slightly higher plasma Vtg concentrations (>1 μg/ml).Plasma Vtg levels in juvenile male carp, which were exposed to effluent from sewage treatment plant for 10-14 days, were higher than those in the unexposed male fish, suggesting that the effluent contained estrogenic compounds at a biological active level. The analyses of the effluent with GC/MS revealed that estradiol- 17β in the effluent is one of the main compounds to increase plasma Vtg level in Koi carp.