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Robert Gale

Publications and source records attributed to Robert Gale.

3 recordsLinked to original sources

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.

Comparative Immunology Reports

Comparison of two estrogen chemically activated luciferase expression cell bioassays to liquid chromatography–mass spectrometry for quantifying estrone in water samples

Chemically activated luciferase expression (CALUX) cell bioassays are popular tools for assessing endocrine activity of chemicals such as certain environmental contaminants. Although activity equivalents can be obtained from CALUX analysis, directly comparing these equivalents to those obtained from analytical chemistry methods can be problematic because of the complexity of endocrine active pathways. We explored the suitability of two estrogen CALUX bioassays (the Organisation for Economic Co-operation and Development–approved VM7Luc4E2 cell bioassay and the VM7LucERβc9 cell bioassay) for quantitation of estrogen. Quadrupole-time of flight ultraperformance liquid chromatography–mass spectrometry (LC/MS) was selected as a comparative method. Regression analysis of measured estrone (E1) calibration samples showed all three methods to be highly predictive of nominal concentrations ( p ≤ 7.5 × 10 –51 ). Extracts of water sampled from laboratory dilutor tanks containing E1 at 0, 20, and 200 ng/L alone and in combination with atrazine were selected to test the quantitative capabilities of the CALUX assays. Process controls (0 and 100 ng E1/L) and a separate E1 standard (10 ng/ml, used to prepare the E1 process control) were also tested. Levels of E1 determined by LC/MS analysis and bioanalytical equivalents (ng E1/L) determined by CALUX analyses were comparable except in certain instances where the samples required dilution prior to CALUX analyses (e.g., the E1 process control and E1 standard). In those instances, measurements by CALUX were slightly but significantly decreased relative to LC/MS. Atrazine had no effect on the ability of either LC/MS or the CALUX bioassays to quantify E1. The present study illustrates the CALUX bioassays as successful in quantifying an estrogen in simple water samples and further characterizes their utility for screening. Environ Toxicol Chem 2023;42:333–339. Published 2022. This article is a U.S. Government work and is in the public domain in the USA.

Environmental Toxicology and Chemistry

Declines in reproductive condition of male largemouth bass (Micropterus salmoides) following seasonal exposure to estrogenic endocrine-disrupting compounds

Reproductive abnormalities, that could lead to possible effects at the population level, have been observed in wild fish throughout the United States, with high prevalence in largemouth bass (LMB; Micropterus salmoides ) and smallmouth bass ( Micropterus dolomieu ). Estrone (E1) and atrazine (ATR) are common environmental contaminants often associated with agricultural land use. 17alpha-ethinylestradiol (EE2) is a contaminant associated with wastewater treatment effluent, and a representative, well-studied estrogen commonly used for fish toxicity testing. Our objective was to assess whether early gonad recrudescence in adult fish was a period of sensitivity for alterations in reproductive condition and function. Adult male LMB were exposed from post-spawning to early gonad recrudescence to either a mixture of E1 (47.9 ng/L) + ATR (5.4 µg/L), or EE2 (2.4 ng/L) in outdoor experimental ponds. Gonad samples were collected from fish just prior to the start of exposure (July), at the end of the exposure period (December), the following spring just prior to spawning (April), and post spawning (May). Gonadosomatic index (GSI) was significantly reduced in E1 + ATR-exposed and EE2-exposed males compared to control at every post-exposure time point. Reduced sperm count and sperm motility were observed in the mixture treatment (E1 + ATR) compared to the control. Sperm motility was also reduced in the EE2 treatment. These data together indicate that estrogenic endocrine-disrupting compounds can lessen the reproductive condition of adult male LMB, and that effects of exposure during early gonad recrudescence can persist at least through the subsequent spawning cycle.

International Journal of Molecular Sciences