We investigated water quality at an advanced water reclamation plant and three conventional wastewater treatment plants using an "ecotoxicity toolbox" consisting of three complementary analyses (chemical analysis, in vitro bioanalysis and in situ biological monitoring), with a focus on endocrine disruption. The in vitro bioassays were chosen to provide an appropriately wide coverage of biological effects relevant to managed aquifer recharge and environmental discharge of treated wastewater, and included bioassays for bacterial toxicity (Microtox), genotoxicity (umuC), photosynthesis inhibition (Max-I-PAM) and endocrine effects (E-SCREEN and AR-CALUX). Chemical analysis of hormones and pesticides using LCMSMS was performed in parallel to correlate standard analytical methods with the in vitro assessment. For two plants with surface water discharge into open drains, further field work was carried out to examine in situ effects using mosquitofish (Gambusia holbrooki) as a bioindicator species for possible endocrine effects. The results show considerable cytotoxicity, phytotoxicity, estrogenicity and androgenicity in raw sewage, all of which were significantly reduced by conventional wastewater treatment. No biological response was detected to RO water, suggesting that reverse osmosis is a significant barrier to biologically active compounds. Chemical analysis and in situ monitoring revealed trends consistent with the in vitro results: chemical analysis confirmed the removal trends observed by the bioanalytical tools, and in situ sampling did not reveal any evidence of endocrine disruption specifically due to discharge of treated wastewater (although other sources may be present). Biomarkers of exposure (in vitro) and effect (in vivo or in situ) are complementary and together provide information with a high level of ecological relevance. This study illustrates the utility of combining multiple lines of evidence in the assessment of water quality.

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Reproductive parameters and accumulation of chlorophenolic compounds by white sucker (Catostomus commersoni) populations exposed to bleached kraft mill effluent (BKME) were studied in the St. Maurice River, Quebec. Compared with unexposed populations from the same waterway, exposed white sucker showed obvious effects of chemical exposure as far as 100 km downstream of the mill, as revealed by a strong induction of ethoxyresorufin O-deethylase (EROD) activity. In males, circulating plasma testosterone levels were the same at all sites, while 11 -ketotestosterone levels were significantly lower at exposed stations. In females, testosterone and 17β-estradiol levels were significantly reduced at the two exposed stations relative to the reference station. Despite different hormone levels during sexual maturation, gonad weight as a proportion of carcass weight was similar at all sites for both sexes during early gonadal development. The relationship between carcass weight and fecundity was more variable at the station immediately downstream of the mill. Biochemical and physiological parameters measured in this study do not allow us to clearly relate perturbations in plasma steroid levels to impaired reproduction as measured by gonad weight and fecundity.

Induction of hepatic ethoxyresorufin‐O‐deethylase (EROD) activity and accumulation of chlorophenolic compounds typical of bleached‐kraft mill effluent (BKME) in fish sampled downstream of a pulp mill on the St. Maurice River, Québec, Canada, provided evidence of chemical exposure to BKME. In comparison, fish sampled over the same distances and in similar habitats in a noncontaminated reference river, the Gatineau River, demonstrated low EROD activity and contamination levels. Accelerated growth of white suckers occurred between 2 and 10 years of age in both rivers at downstream stations relative to upstream stations, suggesting the existence of gradients of nutrient enrichment independent of BKME contamination. The impact of BKME exposure was expressed as reduced investment in reproduction, as revealed by greater length at maturity, reduced gonad size, and more variable fecundity. These effects were not obvious in simple upstream‐downstream comparisons, but became evident when fish from the uncontaminated Gatineau River showed increased gonadal development and reduced age and size at maturity in response to enhanced growth rates.

Immature Atlantic salmon (Salmo salar) were exposed to water accommodated fraction (WAF) of Bass Strait crude oil or to Corexit 9527-dispersed crude oil for 6 days, followed by a depuration period of 29 days. Serum sorbitol dehydrogenase (SDH) levels, indicator of liver damages, remained low during the experiment. Hepatic EROD activity was induced within 2 days following the onset of the exposure in both treatments, and persisted for 2-4 and 4-6 days after transfer to clean sea water in the WAF and dispersed oil treatment, respectively. Naphthalene-type metabolites, determined by fixed-wavelength fluorescence detection, appeared in the bile of the fish with 2 days' delay compared to EROD induction. In both treatments, EROD activity induction and levels of naphthalene-type metabolites in the bile were significantly related. The biliary levels of naphthalene-type metabolites were over 15 times higher in fish exposed to dispersed crude oil relative to fish exposed to the WAF of Bass Strait crude oil. BaP-type metabolites appeared only in the bile of the fish exposed to the WAF, possibly due to BaP-type compounds remaining associated with the dispersant in the water column or to an inhibition of Phase II detoxification enzymes by the dispersant. Bile metabolites as determined by fixed-wavelength fluorescence and EROD induction appear to be sensitive and complementary biomarkers of exposure to PAH.