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.2005 Oct;113(10):1299-307.
doi: 10.1289/ehp.7921.

Assessing the sensitivity of different life stages for sexual disruption in roach (Rutilus rutilus) exposed to effluents from wastewater treatment works

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Assessing the sensitivity of different life stages for sexual disruption in roach (Rutilus rutilus) exposed to effluents from wastewater treatment works

Katherine E Liney et al. Environ Health Perspect.2005 Oct.

Abstract

Surveys of U.K. rivers have shown a high incidence of sexual disruption in populations of wild roach (Rutilus rutilus) living downstream from wastewater treatment works (WwTW), and the degree of intersex (gonads containing both male and female structural characteristics) has been correlated with the concentration of effluent in those rivers. In this study, we investigated feminized responses to two estrogenic WwTWs in roach exposed for periods during life stages of germ cell division (early life and the postspawning period). Roach were exposed as embryos from fertilization up to 300 days posthatch (dph; to include the period of gonadal sex differentiation) or as postspawning adult males, and including fish that had received previous estrogen exposure, for either 60 or 120 days when the annual event of germ cell proliferation occurs. Both effluents induced vitellogenin synthesis in both life stages studied, and the magnitude of the vitellogenic responses paralleled the effluent content of steroid estrogens. Feminization of the reproductive ducts occurred in male fish in a concentration-dependent manner when the exposure occurred during early life, but we found no effects on the reproductive ducts in adult males. Depuration studies (maintenance of fish in clean water after exposure to WwTW effluent) confirmed that the feminization of the reproductive duct was permanent. We found no evidence of ovotestis development in fish that had no previous estrogen exposure for any of the treatments. In wild adult roach that had previously received exposure to estrogen and were intersex, the degree of intersex increased during the study period, but this was not related to the immediate effluent exposure, suggesting a previously determined programming of ovotestis formation.

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Figures

Figure 1
Figure 1
Experimental design for the effluent exposure studies. Bullets indicate sampling points for analytical chemistry (collections of 7-day composite samples of effluent). Blue arrows indicate sampling points for biological analyses. (A) ELS study in which fertilized roach eggs were deployed into graded concentrations of WwTW effluent in mesocosm systems at both sites and maintained until 200 dph (WwTW A) and 300 dph (WwTW B). At both sites at 60 dph (July 2001), 60 fish from each effluent concentration and from controls were transferred to clean water for depuration and were sampled at 300 dph. (B) Postspawning roach studies in which sexually maturing male roach that had previously received exposure to estrogen were exposed to effluent from WwTW A for 4 months and WwTW B for 2 months (adult exposure 2), and another group of males with no previous exposure to estrogen was exposed to effluent from WwTW B for 2 months (adult exposure 1).
Figure 2
Figure 2
Measured concentrations of steroidal estrogens (A) and alkylphenolic chemicals (B) in full-strength (100%) effluent at both study sites during the experiments.
Figure 3
Figure 3
ELS study. (A ) Mean measured concentrations of VTG in whole-body homogenates of fish exposed from fertilization to 200 dph and 300 dph. (B) Percentage of fish with male germ cells and “female-like” reproductive duct exposed to WwTW effluents from fertilization to 200 dph and 300 dph. (C) Percentage of fish with male germ cells and “female-like” ovarian cavities after exposure to WwTW effluent from fertilization to 60 dph followed by depuration in clean water to 300 dph. *p < 0.05, and **p < 0.001 compared with controls.
Figure 4
Figure 4
Gonadal histology of ELS roach after exposure to 100% effluent. (A) Control female. (B) Control male. (C ) Effluent-exposed male. Abbreviations: M, mesentery; O, oogonia; OC, ovarian cavity; PO, primary oocytes; S, spermatogonia; SD, sperm duct. Large arrows indicate points of attachment of the gonad to the mesentery.
Figure 5
Figure 5
Pre/postspawning studies showing effects of exposure to WwTW effluents on (A) condition factor (K ) and (B) GSI. (C ) VTG concentrations in blood plasma in adult male roach. (D) Intersex index of adult fish in exposure 2. *p < 0.05 compared with control.
Figure 6
Figure 6
Pre/postspawning study (adult exposures 1 and 2) showing gonadal histopathology of male and intersex roach. (A andB) Gonads of males with no previous exposure to estrogen. In July (A), the testis was filled with spermatozoa (SZ), and cysts of spermatogonia A (SGA) and spermatogonia B (SGB) were also visible. In September (B), testes of male roach were normal, containing cysts of SGA, SGB, and spermatocytes (SC); there were no obvious differences between the testes of effluent-exposed fish and river water controls. (C–E ) Gonads of males with previous exposure to estrogen. In July, the testis was filled with SZ (C ), and some males were intersex (D). The testis contained SZ, oogonia (O), primary oocytes (PO), and larger oocytes in the cortical alveolus stage (CA). In September (E ), the gonads of these intersex fish contained cysts of SGA, SGB, and SC, together with PO.
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References

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