Evaluating the potential of effluents and wood feedstocks from pulp and paper mills in Brazil, Canada, and New Zealand to affect fish reproduction: chemical profiling and in vitro assessments (original) (raw)
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Environmental toxicology and chemistry, 2016
The authors examined the potential of pulp mill effluent from pulp-producing countries (Canada, Brazil, New Zealand) to affect fish reproduction. Specifically, the estrogenic effects in juvenile rainbow trout (Oncorhynchus mykiss) pulse-exposed to 11 different mill effluent extracts (intraperitoneal injections of solid-phase extraction-dichloromethane nonpolar fraction). The results indicated that effluent extracts were estrogenic in juvenile trout irrespective of the gender, as reflected by increasing level of plasma vitellogenin (VTG; Brazil > New Zealand > Canada). Despite the high variability observed among mills, differences in VTG levels were related to the type of mill process (kraft > elementary chlorine-free kraft > thermomechanical pulping). Moreover, effluent treatments did not appear to significantly decrease VTG induction. A consistent estrogenic effect was observed in those mills that process a combination of feedstocks (softwood and hardwood), with the hig...
Under the Canadian Environmental Effects Monitoring (EEM) program for pulp and paper effluents, the observation of a national response pattern of decreased gonad size and increased fish condition and liver size has triggered a centralized multiagency investigation of cause (IOC) of reproductive impacts in fishes. The purpose of the component of the IOC study presented here is to compare a number of fish bioassays for determining reproductive and reproductive-endocrine effects of a bleached kraft mill effluent. The bleached kraft mill chosen for this study had demonstrated the national response pattern in previous EEM cycles. The bioassays employed to examine reproduction were fathead minnow (Pimephales promelas) 5- and 21-d, mummichog (Fundulus heteroclitus) 25-d, and zebrafish (Danio rerio) 7-d tests, all of which had egg production as the primary reproductive endpoint. Additional bioassays examining reproductive-endocrine endpoints included a 7-d mummichog test, a 7- and a 21-d threespine stickleback (Gasterosteus aculeatus) test, a rainbow trout (Oncorhynchus mykiss) 7-d test, and in vitro sex steroid receptor and plasma protein binding bioassays. The zebrafish and fathead minnow reproductive tests showed significant suppression of egg production at the 100% effluent concentration. Endocrine data derived from the tests showed that this effluent did not impact steroidogenic endpoints at any concentration. Bioassays showed that this effluent i) was capable of eliciting cytochrome P4501A induction at as low as 10% vol/vol effluent, ii) was weakly androgenic at 10% vol/vol, and iii) showed no evidence of in vivo estrogenicity. These results were consistent with in vitro receptor binding assays showing a highly variable level of androgenic equivalents over six months of effluent testing, with little evidence of estrogenic activity. Bioassay results were consistent in that the overall conclusion was that this effluent has only a weak potential to cause reproductive impairment and would likely not do so at environmentally relevant concentrations. Field studies and a fathead minnow lifecycle study conducted concurrently were in agreement with reproductive bioassay results as white sucker exposed in the receiving environment no longer had significantly reduced gonadal development. Overall, this study provided evidence that the laboratory assays evaluated for various reproductive endpoints have potential application for future IOC work.
Environmental Toxicology and Chemistry, 2007
For the last 20 years, studies conducted in North America, Scandinavia, and New Zealand have shown that pulp and paper mill effluents affect fish reproduction. Despite the level of effort applied, few leads are available regarding the factors responsible. Effluents affect reproduction in multiple fish species, as evidenced by decreased gonad size, decreased circulating and gonadal production of reproductive steroids, altered expression of secondary sex characteristics, and decreased egg production. Several studies also have shown that effluent constituents are capable of accumulating in fish and binding to sex steroid receptors/ binding proteins. Studies aimed at isolating biologically active substances within the pulping and papermaking process have provided clues about their source, and work has progressed in identifying opportunities for in-mill treatment technologies. Following comparisons of manufacturing processes and fish responses before and after process changes, it can be concluded that effluent from all types of mill processes are capable of affecting fish reproduction and that any improvements could not be attributed to a specific process modification (because mills normally performed multiple modifications simultaneously). Improved reproductive performance in fish generally was associated with reduced use of molecular chlorine, improved condensate handling, and liquor spill control. Effluent biotreatment has been effective in reducing some effects, but biotreated effluents also have shown no difference or an exacerbation of effects. The role of biotreatment in relation to effects on fish reproduction remains unclear and needs to be resolved.
Environmental Toxicology and Chemistry, 2009
Endocrine disruption (ED) effects due to pulp and paper mill effluents extracts involving different industrial procedures and effluent treatments (nontreated, primary, and secondary treated) were evaluated using immature triploid rainbow trout in a pulse-exposure toxicity experiment. The protocol involved the use of intraperitoneal injection of mill extracts (solid-phase extraction [SPE]) corrected for individual fish weight and included several laboratory standards (steroidal hormones and phytosterols). Biological endpoints at two different levels of biological organization were analyzed (molecular and individual organism). Results indicated that nonsignificant changes were observed in the individual physiological indices represented by condition factor, liver somatic index, and gonad somatic index during the experiment. Significant induction of liver ethoxyresorufin-O-deethylase activity was observed between different effluent treatments and experimental controls. Significant endocrine-disrupting effects at the reproductive level were observed in all effluent treatments involving significant increments in plasma vitellogenin (VTG) levels. Fish exposed to untreated effluent extracts had significantly higher VTG levels compared to fish exposed to primary and secondary treatment effluent extracts, indicating a decrease of the estrogenic effect due to the effluent treatment. The present study has shown that for the Chilean pulp and paper mill SPE extracts evaluated, an endocrine disruption effect was induced in immature triploid rainbow, reaffirming the significant estrogenic effects demonstrated previously in laboratory and field experiments.
Since 1997, a number of approaches (artificial stream exposures, lab bioassays) have been used to identify waste-stream sources of contaminants at the Irving Pulp & Paper Ltd. (IPP) mill, in Saint John, Canada. These studies have shown that chemical recovery condensates have the greatest potential for reducing circulating and gonadal steroids in mummichog (Fundulus heteroclitus), an endemic fish species. A solid phase extraction technique was developed to isolate the hormonally-active substances from the condensates, and a toxicity identification evaluation (TIE) approach was used to gain a better understanding of the chemical characteristics of the active substances. The extract was fractionated by high performance liquid chromatography and the fractions were used in a seven-day bioassay. Mummichog were exposed in static aquaria with daily renewal to either the whole condensate extract, or one of the six fractions at 1 % v/v. Steroid reductions in recent exposures were not as pronounced as had been observed during studies which investigated the overall potency of the extract at the same concentration. As well, differences in responses were observed between the sexes. A dose-response experiment indicated that greater steroid reductions are elicited at 4% v/v in male mummichog. Therefore, the TIE was continued at 4%v/v, however, no steroid reductions were observed in any of the fractions. Some fractions induced increases in plasma testosterone, which had never been observed previously. Ongoing work is focusing on developing an understanding of the extent to which variability in the fish responses are linked to reproductive state and differences in responses between the sexes, while additional work is concentrating on variability in condensate constituents and the fractionation protocol.
Endocrine Disruptor Impacts on Fish From Chile: The Influence of Wastewaters
Frontiers in Endocrinology
Industrial wastewaters and urban discharges contain complex mixtures of chemicals capable of impacting reproductive performance in freshwater fish, called endocrine-disrupting compounds (EDCs). In Chile, the issue was highlighted by our group beginning over 15 years ago, by analyzing the impacts of pulp and paper mill effluents (PPME) in the Biobio, Itata, and Cruces River basins. All of the rivers studied are important freshwater ecosystems located in the Mediterranean region of Central Chile, each with a unique fish biodiversity. Sequentially, we developed a strategy based on laboratory assays, semicontrolled-field experiments (e.g., caging) and wild fish population assessments to explore the issue of reproductive impacts on both introduced and native fish in Chile. The integration of watershed, field, and laboratory studies was effective at understanding the endocrine responses in Chilean freshwater systems. The studies demonstrated that regardless of the type of treatment, pulp ...
Journal of Great Lakes Research, 1996
Our earlier work showed that exposure to bleached kraft pulp mill effluent resulted in a number of reproductive alterations in white sucker including depression of circulating sex steroid hormone levels.. Further studies demonstrated that reductions in the steroid biosynthetic capacity of the ovarian follicle were responsible for these changes. In this study an epidemiological approach was used to evaluate whether fish (white sucker and brown bullhead) exposed to different classes of organic compounds (non-chlorinated pulp mill effluent and polycyclic aromatic hydrocarbons) exhibit comparable alterations in reproductive function (reduced gonadal size, reductions in circulating steroid levels and depressed steroid biosynthetic capacity). The criteria ofprobability, strength of association, and consistency with respect to time were satisfied with respect to these reproductive alterations. The responses observed did not satisfy the criteria of specificity, or consistency with respect to different people and were indeterminate for the criteria of prediction, time order, consistency with respect to different places, species and contaminant as well as for coherence. In conclusion, changes in reproductive performance represents a common response in fish exposed to pulp mill effluent and other organic contaminants, but the biochemical basis for steroid hormone depression in fish associated with organic contaminant exposure is not consistent enough to allow generalizations at this time.
Reproductive steroid responses in fish exposed to pulp mill condensates
2006
Since 1997, a number of approaches (artificial stream exposures, lab bioassays) have been used to identify waste-stream sources of contaminants at the Irving Pulp & Paper Ltd. (IPP) mill, in Saint John, Canada. These studies have shown that chemical recovery condensates have the greatest potential for reducing circulating and gonadal steroids in mummichog (Fundulus heteroclitus), an endemic fish species. A solid phase extraction technique was developed to isolate the hormonally-active substances from the condensates, and a toxicity identification evaluation (TIE) approach was used to gain a better understanding of the chemical characteristics of the active substances. The extract was fractionated by high performance liquid chromatography and the fractions were used in a seven-day bioassay. Mummichog were exposed in static aquaria with daily renewal to either the whole condensate extract, or one of the six fractions at 1 % v/v. Steroid reductions in recent exposures were not as pronounced as had been observed during studies which investigated the overall potency of the extract at the same concentration. As well, differences in responses were observed between the sexes. A dose-response experiment indicated that greater steroid reductions are elicited at 4% v/v in male mummichog. Therefore, the TIE was continued at 4%v/v, however, no steroid reductions were observed in any of the fractions. Some fractions induced increases in plasma testosterone, which had never been observed previously. Ongoing work is focusing on developing an understanding of the extent to which variability in the fish responses are linked to reproductive state and differences in responses between the sexes, while additional work is concentrating on variability in condensate constituents and the fractionation protocol.