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Reproductive Consequences of Exposure to Waterborne Phytoestrogens in Male Fighting Fish Betta splendens

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Abstract

Phytoestrogens are plant compounds that can act as endocrine disruptors in vertebrates. Biologically active levels of phytoestrogens have been found in aquatic habitats near wood pulp and paper mills, biofuel manufacturing plants, sewage-treatment plants, and agricultural fields. Phytoestrogens are known to cause hormonal and gonadal changes in male fish, but few studies have connected these effects to outcomes relevant to reproductive success. In one experiment, we exposed sexually mature male fighting fish Betta splendens to environmentally relevant (1 μg L−1) and pharmacological concentrations (1000 μg L−1) of the phytoestrogen genistein as well as to a positive control of waterborne 17β-estradiol (E2; 1 μg L−1), and a negative control of untreated water. In a second experiment, we exposed male B. splendens to environmentally relevant concentrations (1 μg L−1) of genistein and β-sitosterol singly and in combination as well as to the positive and negative controls. All exposures were 21 days in duration. We measured sex-steroid hormone levels, gonadosomatic index (GSI), sperm concentration and motility, and fertilization success in these fish. We found that exposure to genistein did not affect circulating levels of the androgen 11-ketotestosterone or the estrogen E2 relative to negative-control fish. We also found that neither of the compounds nor their mixture affected GSI, sperm concentration or motility, or fertilization success in exposed fish relative to negative-control fish. However, fish exposed to phytoestrogens showed some evidence of fewer but more motile sperm than fish exposed to the positive control E2. We conclude that sexually mature male B. splendens are relatively immune to reproductive impairments from short-term exposure to waterborne phytoestrogens.

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Acknowledgments

We thank the following people for assistance in the laboratory and for the collection of pilot data that led to this manuscript: Tatiana Butler, Julian Damashek, Genelle Diaz-Silveira, Natalie Ferraiolo, Heather Leonard, Kathy Nieves-Puigdoller, and David Westwood. Thanks to Dante Vargas and M. Monica Giusti for conducting the HPLC analyses of our genistein water samples. Renae Brodie, Beth Jakob, Denise Pope, and two anonymous reviewers offered useful comments on an earlier version of this manuscript. Funding for this research was provided by the Webster Fund of the Department of Biology at Amherst College, the H. Axel Schupf ‘57 Fund for Intellectual Life at Amherst College, HHMI grant 52005107 to Amherst College, and National Science Foundation grant IOS-0725186 to E. D. C.

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  1. Department of Biology, Amherst College, Amherst, MA, 01002, USA

    Louise M. Stevenson, Alexandria C. Brown, Tracy M. Montgomery & Ethan D. Clotfelter

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  1. Louise M. Stevenson
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Correspondence to Ethan D. Clotfelter.

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Louise M. Stevenson and Alexandria C. Brown contributed equally to this work.

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Stevenson, L.M., Brown, A.C., Montgomery, T.M. et al. Reproductive Consequences of Exposure to Waterborne Phytoestrogens in Male Fighting Fish Betta splendens . Arch Environ Contam Toxicol 60, 501–510 (2011). https://doi.org/10.1007/s00244-010-9561-y

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