A field study of hemolymph yolk protein levels in a bivalve (Unio tumidus) and future considerations for bivalve yolk protein as endocrine biomarker

Induction of yolk protein in male fish is a recognized biomarker for estrogenic exposure because the estrogen-dependent induction mechanism is well investigated and there is a clear difference in yolk protein levels of unexposed males and females. Attempts have been made to use induction of bivalve yolk protein as biomarker for estrogenic exposure. However, several biomarker validation criteria have not yet been investigated e.g. an in-depth understanding of the induction mechanism and background variability is needed and reliable detection assays are yet to be developed. To obtain background knowledge about yolk protein levels freshwater bivalves (Unio tumidus) were collected in an uncontaminated Danish lake over the course of a year (33 collection dates). The hemolymph yolk protein concentration of 569 individuals was determined by a species specific enzyme-linked immunosorbent assay (ELISA) and male and female gonadal development cycles were established. The average yolk protein levels of males and females collected at each sampling date overlapped in some periods; the male and female range was 66,946 - 169,692 ng/mL and 88,731 - 681,667 ng/mL, respectively. Because male and female hemolymph yolk protein levels overlap, great care should be taken if yolk protein induction in bivalve hemolymph is considered as endocrine biomarker.