Immunomodulation in adult largemouth bass (Micropterus salmoides) exposed to a model estrogen or mixture of endocrine disrupting contaminants during early gonadal recrudescence

Disease outbreaks, skin lesions, fish kill events, and reproductive abnormalities have been observed in wild populations of Centrarchids in watersheds throughout the United States. Occurrence of synthetic and natural hormones from wastewater treatment plants and livestock operations, pesticides from agricultural land use, and phytoestrogens have been implicated as potential causes of these adverse effects. Our objective was to investigate possible immunomodulation in adult largemouth bass (Micropterus salmoides) in response to a seasonal exposure to environmentally relevant contaminants in outdoor experimental ponds. Exposures included 17α-ethinylestradiol (EE2; 3.6 ng/L) or a binary mixture of endocrine-active substances commonly detected in surface waters, estrone (E1; 85.6 ng/L) and atrazine (ATR; 5.4 µg/L). The 4-month exposure was conducted from July to November. Functional immune responses of anterior kidney-derived leukocytes were evaluated in December in the week following the end of the dosing period, and in the following April, four months after dosing ended and just prior to spawning. Concentrations of EE2 and E1 in the ponds fell below detectable levels in December, but detectable concentrations of ATR (2.9 µg/L) persisted at least through May. For each sampling time, anterior kidney leukocytes were isolated and grown in primary culture for the assessment of zymosan-stimulated respiratory burst and lectin-stimulated mitogenic responses. We observed seasonal differences in respiratory burst stimulation over time and treatment with a significantly greater response in April relative to December. Respiratory burst activity was also significantly greater in April for fish exposed to the E1+ATR relative to control. In April, prior to spawning, we observed a significantly dampened mitogenic response to PHAP (a T cell mitogen) and LPS (a B cell mitogen) in the EE2 treatment relative to control fish. There were no significant differences in mitogenic responses or respiratory burst between sexes. However, there was significantly higher alternative complement pathway hemolytic activity in males compared to females in both the control and E1+ATR treatment groups. Our results demonstrate that environmentally relevant concentrations of contaminants can alter immune function in a socioeconomically important fish species.