Superfeminization as an effect of bisphenol A in \u3ci\u3eMarisa cornuarietis\u3c/i\u3e & Response from Forbes et al. to Oehlmann et al.

[Oehlmann et al.]: Recently, we had the opportunity to read two articles in volume 66 of Ecotoxicology and Environmental Safety. Forbes et al. (2007a, , 2007b) declare “to explore the reproducibility of prior work” showing that bisphenol A (BPA) induces superfeminization in the freshwater snail Marisa cornuarietis (Oehlmann et al., 2000, 2006; Schulte- Oehlmann et al., 2001). Based on the outcome of a toxicity test with the same species, the authors conclude that their results “do not support previous claims of enhanced reproduction in M. cornuarietis in response to exposure to BPA.” We take issue with the declaration of exploring the reproducibility of our challenged work and the validity of the conclusions made by Forbes et al. (2007a, 2007b). Furthermore, we feel the toxicity test is flawed because its experimental design and the selected exposure conditions result in an irresponsiveness of test animals to the tested compound. ... [Forbes et al.]:We thank the editor for giving us the opportunity to respond to Oehlmann et al.’s comments (Oehlmann et al., 2008) on our papers (Forbes et al., 2007a,b). In response to the criticisms of our studies we would first like to emphasize that it was not our aim to exactly repeat the experiments of Oehlmann and colleagues, but rather to produce robust and reproducible results that were statistically valid and that therefore could be used in the risk assessment of bisphenol A (BPA). We are disturbed by the highly speculative suggestion by Oehlmann et al. that we may have studied a different ‘cryptic’ species of Marisa cornuarietis, as there is no evidence to support such suggestion. Furthermore, the snails used in our studies were collected from a documented pristine field site and identified to species by Dr. Sharon File-Emperador from University of Puerto Rico, a recognized snail expert. They were reared for a known number of generations under tightly controlled and documented conditions. In contrast, the snails used in Oehlmann et al.’s studies (e.g., Oehlmann et al., 2006) came from the Dusseldorf Zoo (original site of field collection not indicated) and were periodically supplemented with snails from Florida (location not indicated). There is likewise very little information in the published studies about the culture and husbandry conditions of Oehlmann et al.’s snails, whereas a major portion of our research aimed to identify appropriate husbandry and culture conditions for M. cornuarietis. It is on the basis of such detailed study that our toxicity tests with BPA were conducted

Does Bisphenol A Induce Superfeminization in \u3ci\u3eMarisa cornuarietis\u3c/i\u3e? Part II: Toxicity Test Results and Requirements for Statistical Power Analyses

This study presents results of the effects of bisphenol A (BPA) on adult egg production, egg hatchability, egg development rates and juvenile growth rates in the freshwater gastropod, Marisa cornuarietis. We observed no adult mortality, substantial inter-snail variability in reproductive output, and no effects of BPA on reproduction during 12 weeks of exposure to 0, 0.1, 1.0, 16, 160 or 640 μg/L BPA. We observed no effects of BPA on egg hatchability or timing of egg hatching. Juveniles showed good growth in the control and all treatments, and there were no significant effects of BPA on this endpoint. Our results do not support previous claims of enhanced reproduction in Marisa cornuarietis in response to exposure to BPA. Statistical power analysis indicated high levels of inter-snail variability in the measured endpoints and highlighted the need for sufficient replication when testing treatment effects on reproduction in M. cornuarietis with adequate power

Superfeminization as an effect of bisphenol A in \u3ci\u3eMarisa cornuarietis\u3c/i\u3e & Response from Forbes et al. to Oehlmann et al.

[Oehlmann et al.]: Recently, we had the opportunity to read two articles in volume 66 of Ecotoxicology and Environmental Safety. Forbes et al. (2007a, , 2007b) declare “to explore the reproducibility of prior work” showing that bisphenol A (BPA) induces superfeminization in the freshwater snail Marisa cornuarietis (Oehlmann et al., 2000, 2006; Schulte- Oehlmann et al., 2001). Based on the outcome of a toxicity test with the same species, the authors conclude that their results “do not support previous claims of enhanced reproduction in M. cornuarietis in response to exposure to BPA.” We take issue with the declaration of exploring the reproducibility of our challenged work and the validity of the conclusions made by Forbes et al. (2007a, 2007b). Furthermore, we feel the toxicity test is flawed because its experimental design and the selected exposure conditions result in an irresponsiveness of test animals to the tested compound. ... [Forbes et al.]:We thank the editor for giving us the opportunity to respond to Oehlmann et al.’s comments (Oehlmann et al., 2008) on our papers (Forbes et al., 2007a,b). In response to the criticisms of our studies we would first like to emphasize that it was not our aim to exactly repeat the experiments of Oehlmann and colleagues, but rather to produce robust and reproducible results that were statistically valid and that therefore could be used in the risk assessment of bisphenol A (BPA). We are disturbed by the highly speculative suggestion by Oehlmann et al. that we may have studied a different ‘cryptic’ species of Marisa cornuarietis, as there is no evidence to support such suggestion. Furthermore, the snails used in our studies were collected from a documented pristine field site and identified to species by Dr. Sharon File-Emperador from University of Puerto Rico, a recognized snail expert. They were reared for a known number of generations under tightly controlled and documented conditions. In contrast, the snails used in Oehlmann et al.’s studies (e.g., Oehlmann et al., 2006) came from the Dusseldorf Zoo (original site of field collection not indicated) and were periodically supplemented with snails from Florida (location not indicated). There is likewise very little information in the published studies about the culture and husbandry conditions of Oehlmann et al.’s snails, whereas a major portion of our research aimed to identify appropriate husbandry and culture conditions for M. cornuarietis. It is on the basis of such detailed study that our toxicity tests with BPA were conducted