Abstract
The level of vitellogenin-like (Vg-like) proteins in the hemolymph of the soft-shell clam (Mya arenaria) was indirectly determined using an alkali-labile phosphate (ALP) measurement assay. Biochemical characterization of hemolymph from actively reproducing clams showed the presence of Zn, Ca and of substantial amounts of ALP, all of which are indicative of fish serum vitellogenin, reported to be a Zn and Ca containing glycolipophosphoprotein. ALP assays undertaken on different phosphate-containing compounds (phosphate-buffer saline, diphospholipids and 12,000×g supernatants of Vg-rich trout ovary homogenates) showed little interference by the first two substrates and confirmed the specificity of the ALP assay as an indirect method of determining Vg-like proteins in clam hemolymph. Moreover, clams injected with 17β-estradiol and two known xenoestrogens (nonylphenol and pentachlorophenol) significantly induced hemolymph ALP levels. Field work revealed significant differences in hemolymph ALP levels from clams collected at specific intertidal stations of the Saguenay Fjord during temporal (May to October) and spatial (June) surveys in 1997, which appear to indicate that (anti-)estrogenic chemicals may be present in the Saguenay Fjord. Our preliminary results thus show that this simple and rapid ALP assay holds promise as a biomarker of estrogen disruption with molluscan shellfish as bioindicators. ©1999 John Wiley & Sons, Inc. Environ Toxicol 14: 455–465, 1999
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