Endocrine disruption in western mosquitofish from open and closed aquatic ecosystems polluted by swine farm wastewaters

Swine farm wastewaters (SFWs) are generally discharged either directly into nearby rivers or into fish ponds as a source of fertilizer/food for aquaculture in China. SFWs contain various contaminants including steroid hormones. However, there is an extreme paucity of data on their effects in fish populations. Here we investigated the endocrine disrupting effects of SFWs in G. affinis from 2 rivers (7 sites) and 2 fish ponds (2 sites) receiving SFWs and a reference site in Guangdong Province, China. In this study, a total number of 3078 adult western mosquitofish (Gambusia affinis) were collected and the sex ratio was determined. In addition, secondary sexual characteristics were examined and the transcriptional levels of target genes were analyzed. The results showed the mosquitofish populations had a significant increase in male-to-female ratio from 7 sites (including 2 fish ponds) among the 9 sampling sites. The hemal spines of females were masculinized at most sites while the hemal spines of males were feminized at approximately half of the sites (including 2 fish ponds). Significant reduction in vitellogenin (Vtg) mRNA expression was observed in females from 2 sites (including RS7) while elevated Vtg mRNA expression was noticed in males from 2 sites along the rivers (including RS7). Redundancy analysis showed that androgens in the water samples were closely related with male-to-female ratio in the mosquitofish populations and the masculinized hemal spines of females. The findings from this study demonstrated that discharge of SFWs could result in occurrence of both masculinized females and feminized males in mosquitofish population.