Effects of test media on reproduction in Potamopyrgus antipodarum and of pre-exposure population densities on sensitivity to cadmium in a reproduction test.

Molluscan species can be affected by various anthropogenic substances. Yet, these effects are disregarded in chemical risk assessment as molluscs are unrepresented in standard OECD guidelines. The project "validation of a mollusc reproduction test" (Federal Environment Agency, code 371165417) deals with the development of a test method with the mudsnail Potamopyrgus antipodarum for OECD purposes. In this context, the influence on reproduction of both, different media and varying snail density, has been observed in independent experiments. Further, the impact of density on the outcome of subsequent cadmium (Cd) toxicity in a test has been investigated to refine the existing methodology. First, adult snails were kept in different test media for 12 weeks. Second, snail density was increased for 4 weeks to induce stress. Snails from each density scenario were used for another 4 weeks in a reproduction test at an equal density with 12 μg Cd/L, respectively. Significant differences in reproduction between medium groups were noted after 4 and 8, but not 12, weeks. Further, reproduction was significantly altered by snail density in the beakers but after subsequent 4 weeks at a constant density, no differences were observed between control groups. Cd reduced reproduction and this effect increased with snail density in the pre-exposure period, demonstrating that a previous stress factor may result in increased sensitivity to chemicals and underlines the need for more standardized breeding conditions to minimize effect variations. Based on the outcome of this study, an acclimatization period of 12 weeks must be guaranteed for specimens transferred to another medium. Further, 4 weeks of acclimatization are necessary after density stress. An additional 12 weeks density experiment showed that medium volume in each replicate can be decreased by half to save on chemicals, water and space during tests.