Hybrid ELISAs for vitellogenins of the endangered copper redhorse Moxostoma hubbsi and the shorthead redhorse Moxostoma macrolepidotum (Cypriniformes, catostomidae)

Abstract

The shorthead redhorse (sr) Moxostoma macrolepidotum is endemic in North America, while in Canada the copper redhorse (cr) Moxostoma hubbsi is classified as endangered. A commercial heterologous carp vitellogenin (VTG) sandwich ELISA underestimates VTGs in these species. Hybrid ELISAs, using anti-carp VTG antibodies and purified sr- and cr-VTGs as standards, were validated to measure VTGs in plasma and surface mucus of redhorse species. The hybrid ELISAs showed good precision, sensitivity and accuracy. Following induction with β-estradiol 3-benzoate (E2B), levels of plasma VTG (mean±SEM) in male sr increased from 2.7±1.5 μg/ml to 16.5±2.2 mg/ml. In immature cr, plasma VTG increased to 70.1±13.8 mg/ml after induction, from an initial value of 37.7±51.0 μg/ml. Levels of VTG in mucus from E2B-injected fish were highly correlated ( p<0.0001) with levels in plasma, but not in a non-induced mature female. Hybrid assays are a good compromise between homologous and heterologous assays, especially for small-bodied or endangered fish species. The performance of the hybrid assays strongly suggests they could be used to determine exposure of these species to estrogenic contaminants and contribute to restoration efforts of the copper redhorse.