Disturbances in reproduction and expression of steroidogenic enzymes in aquatic invertebrates exposed to components of the herbicide Roundup

Abstract

Exposure of organisms to environmental contaminants is a growing concern. We have investigated the effects of the individual active ingredients of the herbicide Roundup (glyphosate and diquat dibromide [DD]) since Roundup causes alterations in reproduction, mortality, and development in the aquatic snail Lymnaea palustris. Snails chronically treated with elevated but ecologically relevant levels of DD exhibit reduction in fecundity ( p < 0.05), while fecundity in glyphosate-treated snails is comparable to or exceeds control levels. To investigate a possible mechanism for the reproductive disturbance, we monitored levels of steroid acute regulatory (StAR) protein in whole snails and observed a correlation in StAR protein decrease with treatment with Roundup, glyphosate, or DD. We detect StAR in organs where steroid biosynthesis occurs (ovotestis, brain, kidney); StAR protein is reduced following chronic exposure to Roundup, glyphosate, or DD ( p < 0.01). Estradiol and testosterone concentrations in hemolymph were measured by enzyme-linked immunosorbent assay following 3-week exposure of snails to 3.5 mg/L glyphosate or 140 µg/L DD. Testosterone levels decrease in DD-treated groups ( p < 0.05); a trend of lower testosterone is also observed in glyphosate-treated groups ( p > 0.05). Estradiol concentration is greater than or equal to control levels in glyphosate, but decreased in DD ( p < 0.05). Because of its role in the conversion of testosterone to estradiol, we monitored abundance of aromatase and observed a reduction ( p < 0.05) in DD-treated snails (consistent with the drop in fecundity and estradiol levels) and a comparable level to control in glyphosate-treated snails (consistent with their high fecundity and estradiol levels). Although the toxicity of commercially-available Roundup to aquatic animals may have many contributing factors including its inactive surfactant, the constituent of Roundup associated with the greatest reproductive disturbances and observed developmental abnormalities of offspring is DD. This study details the analysis of particular herbicide constituents and their effect on specific targets in the reproductive pathway.