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Metal bioavailability in aquatic organisms is known to be influenced by various water chemistry parameters. The present study examined the influence of alkalinity and natural organic matter (NOM) on tissue-specific metal accumulation and reproductive performance of fathead minnows (Pimephales promelas) during environmentally relevant chronic exposures to a metal mine effluent (MME). Sodium bicarbonate (NaHCO3) or NOM (as commercial humic acid) were added to a Canadian MME [45 percent process water effluent (PWE)] in order to evaluate whether increases in alkalinity (3–4 fold) or NOM (~1.5–3mg/L dissolved organic carbon) would reduce metal accumulation and mitigate reproductive toxicity in fathead minnows during a 21-day multi-trophic exposure. Eleven metals (barium, boron, cobalt, copper, lithium, manganese, molybdenum, nickel, rubidium, selenium, and strontium) were elevated in the 45 percent PWE relative to the reference water. Exposure to the unmodified 45 percent PWE resulted in a decrease of fathead minnow egg production (~300 fewer eggs/pair) relative to the unmodified reference water, over the 21-day exposure period. Water chemistry modifications produced a modest decrease in free ion activity of some metals (as shown by MINTEQ, Version 3) in the 45 percent PWE exposure water, but did not alter the metal burden in the treatment-matched larval Chironomus dilutus (the food source of fish during exposure). The tissue-specific metal accumulation increased in fish exposed to the 45 percent PWE relative to the reference water, irrespective of water chemistry modifications, and the tissue metal concentrations were found to be similar between fish in the unmodified and modified 45 percent PWE (higher alkalinity or NOM) treatments. Interestingly however, increased alkalinity and NOM markedly improved fish egg production both in the reference water (~500 and ~590 additional eggs/pair, respectively) and 45 percent PWE treatments (~570 and ~260 additional eggs/pair, respectively), although fecundity over 21 day exposure consistently remained lower in the 45 percent PWE treatment groups relative to the treatment-matched reference groups. Collectively, these findings suggest that metal accumulation caused by chronic 45 percent PWE exposure cannot solely explain the reproductive toxicity in fish, and decrease in food availability (decrease in C. dilutus abundance in 45 percent PWE exposures) might have played a role. In addition, it appears that NaHCO3 or humic acid mitigated reproductive toxicity in fish exposed to 45 percent PWE by their direct beneficial effects on the physiological status of fish. [Copyright &y& Elsevier]

Abstract: Metal mine effluents can impact fish in the receiving environment via both direct effects from exposure as well as indirect effects via food web. The main objective of the present study was to assess whether an indirect effect such as reduced food (prey) availability could influence metal accumulation and reproductive capacity in fish during chronic exposure to a metal mine effluent. Breeding pairs of fathead minnows (Pimephales promelas) were exposed to either reference water (RW) or an environmentally relevant metal mine effluent [45 percent process water effluent (PWE)] for 21 days and fed either low food quantities [LF (a daily ration of 6–10 percent body weight)] or normal food quantities [NF (a daily ration of 20–30 percent body weight)] in artificial stream systems. Fish in RW treatments were fed Chironomus dilutus larvae cultured in RW (Treatments: RW-NF or RW-LF), while fish in PWE treatments were fed C. dilutus larvae cultured in PWE (Treatments: PWE-NF or PWE-LF). Tissue-specific (gill, liver, gonad and carcass) metal accumulation, egg production, and morphometric parameters in fish were analyzed. Fathead minnows that were exposed to LF rations had significantly smaller body, gonad and liver sizes, and were in a relatively poor condition compared to fathead minnows exposed to NF rations, regardless of the treatment water type (RW or PWE) (two-way ANOVA; p<0.05). Although elevated concentrations of copper, nickel, rubidium, selenium, and thallium were recorded in C. dilutus cultured in PWE, only the concentrations of rubidium, selenium and thallium increased in tissues of fish in PWE treatments. Interestingly though, despite the greater abundance of metal-contaminated food in the PWE-NF treatment, tissue metal accumulation pattern were almost similar between the PWE-NF and PWE-LF treatments, except for higher liver barium, cobalt and manganese concentrations in the latter treatment. This indicated that a higher food ration could help reduce the tissue burden of at least some metals and thereby ameliorate the toxicity of metal-mine effluents in fish. More importantly, cumulative egg production in fish was found to be lowest in the PWE-LF treatment, whereas fish egg production in the PWE-NF treatment was not impacted. Overall, these findings suggest that decreased food abundance could have a greater impact than metal accumulation in target tissues on the reproductive capacity of fish inhabiting metal-mine effluent receiving environments. [Copyright &y& Elsevier]

This study illustrates the use of a mesocosm approach for assessing the independent effects of three treated metal mine effluents (MME) discharging into a common receiving environment and regulated under the same regulation. A field-based, multi-trophic artificial stream study was conducted in August 2008 to assess the effects of three metal mining effluents on fathead minnow (Pimephales promelas) in a 21-day reproduction bioassay. The nature of the approach allowed for assessment of both dietary and waterborne exposure pathways. Elements (e.g. Se, Co, Cl, Cu, Fe) were analyzed in several media (water, sediments) and tissues (biofilm, Chironomus dilutus, female fathead minnow (FHM) body, ovary, liver, gills). Significant increases in metal and micronutrient concentrations were observed in the water and biofilm tissues in all MME treatments [20% surface water effluent (SWE), 30% mine water effluent (MWE), and 45% process water effluent (PWE)], compared to reference. However, copper was the only element to significantly increase in the sediments when exposed to PWE. Co and Ni increased significantly in C. dilutus tissues in SWE (1.4- and 1.5-fold, respectively), Cu and Se also increased in chironomid tissues in PWE (5.2- and 3.3-fold, respectively); however, no significant increases in metals or micronutrients occurred in chironomid tissues when exposed to MWE compared to reference. There were no significant increases in metal concentrations in female FHM tissues (body, liver, gonads, gills) in any of the treatments suggesting that metals were either not bioavailable, lost from the females via the eggs, or naturally regulated through homeostatic mechanisms. Cumulative number of eggs per female per day increased significantly (∼127%) after exposure to SWE and decreased significantly (∼33%) after exposure to PWE when compared to reference. Mean total number of days to hatch was reduced in PWE compared to reference. This study shows the importance of isolating treatment streams in cumulative discharge environments to assess aquatic effects due to the different nature of the effluents. [Copyright &y& Elsevier]

Juvenile fathead minnows (Pimephales promelas) were exposed from 10 to 100 days post-hatch (dph) to metal mining effluent under simulated summer (20°C and 16/8 light/dark) or winter (4°C and 8/16 light/dark) conditions in order to test the winter stress syndrome hypothesis. The condition factor of fish at 100dph was similar between seasons, and was greater in the summer effluent exposure group compared to the summer control. Whole body triglycerides were lower in the effluent exposure group compared to control under winter conditions, but not summer. Whole body burdens of several trace metals, notably Cu, Rb and Se, were greater in the effluent exposed fish at 100dph and displayed seasonal differences in bioaccumulation. Fathead minnows were also exposed from 10 to 100dph to graded concentrations of ammonia (0.02–0.40mg unionized NH3/L) under summer conditions in order to investigate potential effects of this component of the effluent. There was no effect of ammonia exposure on growth parameters, but an increase in whole body triglycerides was observed at the highest exposure. The seasonal differences in bioaccumulation and biological responses suggest that winter conditions may be an important modifying factor in aquatic toxicological studies. [Copyright &y& Elsevier]

Abstract: Junction Creek in Sudbury, Ontario, Canada receives effluent from three metal mining effluents (MMEs), as well as urban run-off and municipal sewage treatment plant (STP) discharges. The present study examined organismal and sub-organismal end-points in prespawning fathead minnow (FHM; Pimephales promelas) and creek chub (CC; Semotilus atromaculatus) collected in May 2004 from Junction Creek. Metal body burdens of Cd, Cu, Rb, Se, and Sr increased in a gradient down Junction Creek in both species. In general, male fish had lower body weights downstream of effluents, while females were unchanged or larger compared to reference fish. Livers were larger in FHM of both sexes downstream of STP and in CC downstream of two MMEs compared to reference fish, while gonad weight was unchanged. The most consistent finding was delayed development, increased cell death and increased eosinophilia in FHM testes collected downstream of all effluents, while female FHM and both sexes of CC gonads were relatively unaffected. In conclusion, the results of the current study in Junction Creek suggest that male FHM reproduction may be sensitive to multiple effluents, but failed to clearly establish whether MME or STP effluents were causative agents over urban inputs or historical contamination. [Copyright &y& Elsevier]

Past studies at a bleached kraft pulp mill located in Saint John, NB, Canada have shown that chemical recovery condensates significantly depress circulating and gonadal steroids in mummichog (Fundulus heteroclitus), an endemic fish species. In the present study, compounds were extracted from the condensates, and a toxicity identification evaluation (TIE) was conducted to characterize the hormonally active substances (HASs) in the condensates. Extracts were fractionated by high performance liquid chromatography (HPLC) and mummichog were exposed to the fractions in a 7-day bioassay. Plasma testosterone was measured for each sex following exposure. Responses in fish exposed to the whole extract at 1% v/v were not as pronounced as had been observed previously (female plasma testosterone was reduced by 16% in the current study compared to 75% previously). Dose–response experiments showed an exposure concentration of 4% v/v was required to elicit significant plasma steroid reductions. Despite these responses, individual condensate fractions actually increased steroid levels in mummichog, which suggests that multiple HASs may need to act synergistically or additively to elicit effects, and if separated, the compounds may have different hormonal activity. The HASs in question caused a reduction in male gonad size at 4% v/v, and have sex-dependent mechanisms of action (males were more responsive to exposure than females). [Copyright &y& Elsevier]