Exposure to 17α-ethynylestradiol causes dose and temporally dependent changes in intersex, females and vitellogenin production in the Sydney rock oyster

Although mounting evidence suggests exposure to estrogenic contaminants increases vitellogenin production in molluscs, demonstration of dose–response relationships and knowledge of the temporal nature of the vitellogenin response with continual exposure is currently lacking for biomarker utility. To address this knowledge gap, adult Sydney rock oysters, Saccostrea glomerata, were exposed to a range of environmentally relevant concentrations of 17α-ethynylestradiol (EE2) (0, 6.25, 12.5, 25 or 50 ng/l) in seawater under laboratory conditions. Vitellogenin induction and gonadal development was assessed following 4, 21 and 49 days exposure to EE2. Vitellogenin was found to increase in a dose dependent manner with EE2 exposure for females (4 and 49 days) and males (4 and 21 days). Histological examination of gonads revealed a number of individuals exhibited intersex (ovotestis) in 50 ng/l EE2 (after 21 days) and in 6.25 and 12.5 ng/l EE2 (after 49 days). Furthermore, a significant shift towards females was observed following 49 days exposure at 50 ng/l EE2 suggesting estrogenic exposure is capable of facilitating a progression for protandric males from male-intersex-female gametal status. Increases in female vitellogenin (4 days) were predictive of later increases in female developmental stages at 21 days and increases in oocyte area following 49 days. Male vitellogenin (4 days) was predictive of decreased male percentages and lower male developmental stages at 49 days. Vitellogenin in S. glomerata is a predictive biomarker of estrogenic exposure and effect if sampled soon after exposure and at the commencement of a gonadal development cycle.

[1]  R. Huggett,et al.  Biomarkers: Biochemical, Physiological, and Histological Markers of Anthropogenic Stress , 1992 .

[2]  F. Gagné,et al.  Determination of vitellogenin‐like properties in Mya arenaria hemolymph (Saguenay Fjord, Canada): A potential biomarker for endocrine disruption , 1999 .

[3]  C. Vane,et al.  Partitioning, bioavailability and effects of oestrogens and xeno-oestrogens in the aquatic environment , 2005, Journal of the Marine Biological Association of the United Kingdom.

[4]  F. Gagné,et al.  Evaluation of estrogenic effects of municipal effluents to the freshwater mussel Elliptio complanata. , 2001, Comparative biochemistry and physiology. Toxicology & pharmacology : CBP.

[5]  T. Roughley The life history of the Australian oyster (Ostrea commercialis) , 1933 .

[6]  D. Morritt,et al.  Long-term and transgenerational effects of nonylphenol exposure at a key stage in the development of Crassostrea gigas. Possible endocrine disruption? , 2003 .

[7]  G. Kraak,et al.  Functional characterization of estrogen receptor subtypes, ERα and ERβ, mediating vitellogenin production in the liver of rainbow trout , 2007 .

[8]  M. Madden,et al.  Biomarkers of Exposure , 1999 .

[9]  M. Ortiz-Zarragoitia,et al.  Biomarkers of Exposure and Reproduction-Related Effects in Mussels Exposed to Endocrine Disruptors , 2006, Archives of environmental contamination and toxicology.

[10]  M. Osada,et al.  Vitellogenin synthesis in the ovary of scallop, Patinopecten yessoensis: Control by estradiol-17 beta and the central nervous system. , 2003, Journal of experimental zoology. Part A, Comparative experimental biology.

[11]  Edwin J. Routledge,et al.  Identification of Estrogenic Chemicals in STW Effluent. 1. Chemical Fractionation and in Vitro Biological Screening , 1998 .

[12]  D. Howard,et al.  Histological techniques for marine bivalve mollusks , 1983 .

[13]  J. Rotchell,et al.  Laboratory exposure to 17beta-estradiol fails to induce vitellogenin and estrogen receptor gene expression in the marine invertebrate Mytilus edulis. , 2006, Aquatic toxicology.

[14]  G. Aherne,et al.  The relevance of the presence of certain synthetic steroids in the aquatic environment , 1989, The Journal of pharmacy and pharmacology.

[15]  M. Asif Hermaphroditism and sex reversal in the four common oviparous species of oysters from the coast of Karachi , 1979, Hydrobiologia.

[16]  F. Gagné,et al.  Health status of Mya arenaria bivalves collected from contaminated sites in Canada (Saguenay Fjord) and Denmark (Odense Fjord) during their reproductive period. , 2006, Ecotoxicology and environmental safety.

[17]  Yuji K. Takahashi,et al.  Oyster estrogen receptor: cDNA cloning and immunolocalization. , 2007, General and comparative endocrinology.

[18]  M. Servos,et al.  Behavior and occurrence of estrogens in municipal sewage treatment plants--I. Investigations in Germany, Canada and Brazil. , 1999, The Science of the total environment.

[19]  A. Hamza-Chaffai,et al.  Endocrine-related reproductive effects in molluscs. , 2008, Comparative biochemistry and physiology. Toxicology & pharmacology : CBP.

[20]  W. O'Connor,et al.  Effects of 4-nonylphenol and 17alpha-ethynylestradiol exposure in the Sydney rock oyster, Saccostrea glomerata: Vitellogenin induction and gonadal development. , 2008, Aquatic toxicology.

[21]  P. Beninger,et al.  The use of physiological condition indices in marine bivalve aquaculture , 1985 .

[22]  M. Marin,et al.  Can 17-β estradiol induce vitellogenin-like proteins in the clam Tapes philippinarum? , 2008, Environmental toxicology and pharmacology.

[23]  W. Langston,et al.  Feminisation of male clams Scrobicularia plana from estuaries in Southwest UK and its induction by endocrine-disrupting chemicals , 2007 .

[24]  Y. Osawa,et al.  Gonadal Estrogen Profile and Immunohistochemical Localization of Steroidogenic Enzymes in the Oyster and Scallop during Sexual Maturation , 1997 .

[25]  Yasuko Nakamura,et al.  Effect of Steroid on Oyster-III , 1968 .

[26]  M. Mulcahy,et al.  Gametogenesis of the oyster Crassostrea gigas in southern Ireland , 1999, Journal of the Marine Biological Association of the United Kingdom.

[27]  M. Marin,et al.  Can 4-nonylphenol induce vitellogenin-like proteins in the clam Tapes philippinarum? , 2005, Environmental research.

[28]  J. W. Thornton,et al.  The Octopus vulgaris estrogen receptor is a constitutive transcriptional activator: evolutionary and functional implications. , 2006, Endocrinology.

[29]  Tohru Suzuki,et al.  Changes in vitellin during oogenesis and effect of estradiol-17β on vitellogenesis in the Pacific oyster Crassostrea gigas , 1998 .

[30]  S. Papa,et al.  Morphological and biochemical characterization of mitochondria in Torpedo red blood cells. , 2001, Comparative Biochemistry and Physiology Part B Comparative Biochemistry.

[31]  P. Dinamani Reproductive cycle and gonadial changes in the New Zealand rock oyster Crassostrea glomerata , 1974 .

[32]  J. Nell,et al.  Studies on triploid oysters in Australia: XII. Gonad discolouration and meat condition of diploid and triploid Sydney rock oysters (Saccostrea commercialis) in five estuaries in New South Wales, Australia , 1999 .

[33]  F. Gagné,et al.  Alteration of the biochemical properties of female gonads and vitellins in the clam Mya arenaria at contaminated sites in the Saguenay Fjord. , 2002, Marine environmental research.

[34]  P. Shin,et al.  Induction, adaptation and recovery of biological responses: implications for environmental monitoring. , 2005, Marine pollution bulletin.

[35]  F. Gagné,et al.  Organic alkali‐labile phosphates in biological materials: A generic assay to detect vitellogenin in biological tissues , 2000 .

[36]  Peter Scanes,et al.  ‘Oyster watch’: Monitoring trace metal and organochlorine concentrations in Sydney's coastal waters , 1996 .

[37]  T. Kayano,et al.  Molecular Characterization of a cDNA Encoding Putative Vitellogenin from the Pacific Oyster Crassostrea gigas , 2003, Zoological science.

[38]  R. Lafont,et al.  Steroids in aquatic invertebrates , 2007, Ecotoxicology.

[39]  G. Maguire,et al.  Studies on triploid oysters in Australia. VI. Gonad development in diploid and triploid Sydney rock oysters Saccostrea commercialis (Iredale and Roughley) , 1996 .

[40]  T. Miyata,et al.  Tissue preferential expression of estrogen receptor gene in the marine snail, Thais clavigera. , 2006, General and comparative endocrinology.

[41]  Christian Blaise,et al.  Vitellogenin as a biomarker of exposure to estrogenic compounds in aquatic invertebrates: a review. , 2008, Environment international.

[42]  M. Marin,et al.  Effects of 4-nonylphenol exposure in mussels (Mytilus galloprovincialis) and crabs (Carcinus aestuarii) with particular emphasis on vitellogenin induction. , 2008, Marine pollution bulletin.

[43]  Sandra E. Shumway,et al.  Scallops : biology, ecology, and aquaculture , 1991 .

[44]  Ximing Guo,et al.  GENETIC DETERMINANTS OF PROTANDRIC SEX IN THE PACIFIC OYSTER, CRASSOSTREA GIGAS THUNBERG , 1998, Evolution; international journal of organic evolution.

[45]  Bent Halling-Sørensen,et al.  Fate of estrogens in a municipal sewage treatment plant. , 2003, Environmental science & technology.

[46]  J. Francis Statistica for Windows , 1995 .

[47]  D. Pampanin,et al.  Stress biomarkers and alkali-labile phosphate level in mussels (Mytilus galloprovincialis) collected in the urban area of Venice (Venice Lagoon, Italy). , 2005, Environmental pollution.

[48]  R. Pipe Ultrastructural and cytochemical study on interactions between nutrient storage cells and gametogenesis in the mussel Mytilus edulis , 1987 .

[49]  A. D. Vethaak,et al.  Analysis and occurrence of estrogenic hormones and their glucuronides in surface water and waste water in The Netherlands. , 1999, The Science of the total environment.

[50]  C. Gagnon,et al.  Exposure of caged mussels to metals in a primary-treated municipal wastewater plume. , 2006, Chemosphere.