Abstract-The androgenic potential of a New Zealand pulp and paper mill effluent was measured by applying a combinatioll of in vitro and in vivo bioassays with mosquitofish (Gambusia a!finis) and goldfish (Carassius aurarus). The in vivo method assessed the rate of gonopodial development (masculinization) and alterations from normal reproductive behavior in adult female mosqllitofish exposed for 21 d to lIntreated 01' secondary-treated pulp mill effluent. A second in vivo mosquitofish exposure tested the effect of gl ass-fiber (type C) fi Itration of secondary-treated effIlient on rates of expression of the same endpoints. Extractable organics analyses of effluents and extracts thereof were conducted. Mosqllitofish demonstrated significant masculinization on exposure to either treated 01' untreated effluent; the frequency of gonopodial development was reduced with efiluent secondary-treatment. Male mating behavior was observed in the masculinized adult females. Glass-fiber (type F) filtration of the treated effluent eliminated the masclilinizing effect, suggesting that the bioactive compounds were associated with the suspended solids. The in vitra method measlired the binding of compounds within a treated thermomechanicallbleached kraft cffluent extract to androgen receptors contained in goldfish testis cytosol. Exposure to extracts of either the particulate (glass-fiber filtered) 01' the dissolved organic fraction of the efflllent produced significant binding (as indicated by the displacement of radiolabeled testosterone) to the androgen reeeptor in goldfish gonadal tissue. Thus, the dissolved organics extract of the treated effluent contained compounds androgenic to goldfish in vitro but not to mosquitofish in vivo. The combined in vitra and in vivo data suggest that the effluent in question could exert effects on the reproductive physiology of fishes through an androgenic mechanism. The androgenic compounds androstenedione and testosterone were not detected in the extracts used far the in vitro component of this study.
The androgenic potential of a New Zealand pulp and paper mill effluent was measured by applying a combination of in vitro and in vivo bioassays with mosquitofish (Gambusia affinis) and goldfish (Carassius auratus). The in vivo method assessed the rate of gonopodial development (masculinization) and alterations from normal reproductive behavior in adult female mosquitofish exposed for 21 d to untreated or secondary-treated pulp mill effluent. A second in vivo mosquitofish exposure tested the effect of glass-fiber (type C) filtration of secondary-treated effluent on rates of expression of the same endpoints. Extractable organics analyses of effluents and extracts thereof were conducted. Mosquitofish demonstrated significant masculinization on exposure to either treated or untreated effluent; the frequency of gonopodial development was reduced with effluent secondary-treatment. Male mating behavior was observed in the masculinized adult females. Glass-fiber (type F) filtration of the treated effluent eliminated the masculinizing effect, suggesting that the bioactive compounds were associated with the suspended solids. The in vitro method measured the binding of compounds within a treated thermomechanical/bleached kraft effluent extract to androgen receptors contained in goldfish testis cytosol. Exposure to extracts of either the particulate (glass-fiber filtered) or the dissolved organic fraction of the effluent produced significant binding (as indicated by the displacement of radiolabeled testosterone) to the androgen receptor in goldfish gonadal tissue. Thus, the dissolved organics extract of the treated effluent contained compounds androgenic to goldfish in vitro but not to mosquitofish in vivo. The combined in vitro and in vivo data suggest that the effluent in question could exert effects on the reproductive physiology of fishes through an androgenic mechanism. The androgenic compounds androstenedione and testosterone were not detected in the extracts used for the in vitro component of this study.
The androgenic potential of a New Zealand pulp mill effluent (PME) and a Canadian PME was assessed along with a New Zealand sewage treatment plant effluent (STP) using a combination of in vivo and in vitro methods. The in vitro methods included: (1) a fish-based androgen receptor binding assay, (2) a fish-based aromatase inhibition activity assay, and (3) an analysis of gonadal sex steroid levels in exposed female mosquitofish ( Gambusia affinis ) ovaries by radioimmunoassay. The in vivo method included a quantifiable analysis of anal fin ray length for female mosquitofish exposed to the effluents. Effluent extracts for the Canadian PME and New Zealand STP were found to have low in vitro androgenic potential compared to upstream reference extracts. All effluent extracts (Canadian PME, New Zealand PME and STP) showed a low degree of in vitro aromatase inhibition potential compared to upstream reference extracts. In vivo analysis showed no androgenic potential of the New Zealand PME and STP. The in vitro androgen receptor assay and in vivo mosquitofish bioassay did show androgenic responses for androstenedione (AD) and 1,4-androsta-diene-3,17-dione (ADD), which are two products of the microbial conversion of β-sitosterol (a plant sterol commonly found in PME) by Mycobacterium smegmatis . Also, the potential of the mosquitofish bioassay to determine anti-androgenic effects in effluents was demonstrated.
The androgenic potential of a New Zealand pulp mill effluent (PME) and a Canadian PME was assessed along with a New Zealand sewage treatment plant effluent (STP) using a combination of in vivo and in vitro methods. The in vitro methods included: (1) a fish-based androgen receptor binding assay, (2) a fish-based aromatase inhibition activity assay, and (3) an analysis of gonadal sex steroid levels in exposed female mosquitofish ( Gambusia affinis ) ovaries by radioimmunoassay. The in vivo method included a quantifiable analysis of anal fin ray length for female mosquitofish exposed to the effluents. Effluent extracts for the Canadian PME and New Zealand STP were found to have low in vitro androgenic potential compared to upstream reference extracts. All effluent extracts (Canadian PME, New Zealand PME and STP) showed a low degree of in vitro aromatase inhibition potential compared to upstream reference extracts. In vivo analysis showed no androgenic potential of the New Zealand PME and STP. The in vitro androgen receptor assay and in vivo mosquitofish bioassay did show androgenic responses for androstenedione (AD) and 1,4-androsta-diene-3,17-dione (ADD), which are two products of the microbial conversion of β-sitosterol (a plant sterol commonly found in PME) by Mycobacterium smegmatis . Also, the potential of the mosquitofish bioassay to determine anti-androgenic effects in effluents was demonstrated.
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