Evaluating the potential of effluent extracts from pulp and paper mills in Canada, Brazil, and New Zealand to affect fish reproduction: Estrogenic effects in fish (original) (raw)
Related papers
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.
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.
Aquatic Toxicology, 2010
Wood extractives present in pulp and paper mill effluents may cause reproductive disturbances in fish. A chronic-exposure toxicity experiment using immature rainbow trout (Oncorhynchus mykiss) was conducted in order to assess the endocrine disrupting effects of two Chilean pulp and paper mill specific extracts (solid phase extraction, SPE) obtained from primary and secondary treated effluents. The (anti)estrogenic potencies and toxicity of the wood extractives regularly present in pulp mill effluent such as dehydroabietic acid (DHAA), -sitosterol (BS), and model estrogen 17-estradiol (E2) were evaluated by analysis of plasma vitellogenin (VTG) levels, gonadal somatic index (GSI) and liver ethoxyresorufin-O-deethylase (EROD) activity, respectively. The protocol involved the use of multiple intra-peritoneal injections (1 injection every 7 days for a total exposure period of 28 days). Analysis of variance/covariance, demonstrated no differences associated with fish gender other than GSI. The phytosterol BS, E2 and both pulp mill effluent extracts showed significant inductions of EROD and increased VTG levels after 4, 7, 14, 21 and 28 of exposure. While fish injected with secondary treated effluent extract showed a delayed induction in VTG levels compared to primary effluent injected fish, no effects on VTG and EROD levels were observed in DHAA injected fish. Moreover simultaneous injection of DHAA + E2 reduced the VTG levels found in E2 injected fish, indicating a potential indirect anti-estrogenic effect of this resin acid. The results of this study indicate that Chilean pulp and paper mill effluent extracts are estrogenic in rainbow trout males and females.
Environmental science & technology, 2012
This study investigates factors affecting reproduction in fish exposed to pulp and paper mill effluents by comparing effluents from countries with varying levels of documented effects. To explore the hypothesis of wood as a common source of endocrine disrupting compounds, feedstocks from each country were analyzed. Analyses included in vitro assays for androgenic activity (binding to goldfish testis androgen receptors), estrogenic activity (yeast estrogen screen), and neurotransmitter enzyme inhibition (monoamine oxidase and glutamic acid decarboxylase). Chemical analyses included conventional extractives, known androgens, and gas chromatograph index (GCI) profiles. All effluents and wood contained androgenic activity, particularly in nonpolar fractions, although known androgens were undetected. Effluents with low suspended solids, having undergone conventional biotreatment had lower androgenic activities. Estrogenic activity was only associated with Brazilian effluents and undetected in wood. All effluents and wood inhibited neurotransmitter enzymes, predominantly in polar fractions. Kraft elemental chlorine free mills were associated with the greatest neurotransmitter inhibition. Effluent and wood GCI profiles were correlated with androgenic activity and neurotransmitter enzyme inhibition. Differences in feedstock bioactivities were not reflected in effluents, implying mill factors mitigate bioactive wood components. No differences in bioactivities could be discerned on the basis of country of origin, thus we predict effluents in regions lacking monitoring would affect fish reproduction and therefore recommend implementing such programs.
Aquatic Toxicology, 2010
We evaluated plasma testosterone (T) and 17-estradiol (E2) levels and ovarian aromatase CYP19a gene expression following a single intraperitoneal injection of Chilean pulp and paper mill effluent extracts into juvenile triploid rainbow trout. Fish injected with untreated effluent extracts had increased plasma T after 4 days, while plasma E2 concentration was increased in fish injected with both primary and secondary treated effluent extracts at the same sampling period. Ovarian CYP19a gene expression as measured by qRT-PCR was significantly induced in fish injected with the untreated, primary and secondary treated pulp and paper mill effluent extracts. Similar induction of CYP19a expression was found in fish injected with the androgens androstenedione (ADD) and T. A Principal Component Analysis (PCA) was conducted in order to identify structure in relationships between all measured variables and identifying which factors were most responsible for the variance observed within the plasma steroid levels, upregulation of ovarian CYP19a gene expression and the final estrogenic effect of increased plasma VTG levels. This analysis indicated a cluster correlation between plasma T levels and CYP19a gene expression (Factor 1, explaining 27.2% of total variance), a cluster including condition factor and liver somatic index (Factor 2, explaining 17.3%) and an additional cluster including plasma E2 and vitellogenin levels (Factor 3, explaining an additional 15.8%). The present results indicate that Chilean pulp and paper mill effluent extracts cause estrogenic effects in triploid rainbow trout. These effects could be related to the compounds present in the effluent that act as estrogen receptor agonists, or that induce changes leading to increased amounts of endogenous estrogens, reflected by increased E2 levels and induced aromatase expression/activity.
Aquatic Toxicology, 2015
Pine and Eucalyptus pulp and paper mill effluents Bioassay In situ Onchorhynchus mykiss Intersex Vitellogenin EROD a b s t r a c t Pulp and paper mill effluents (PPMEs) have been shown to increase gonad size, cause early maturation, and disrupt hormone functions in native and non-native Chilean fish. In this study, we assessed reproductive (plasma vitellogenin; VTG, gonad development) and metabolic (ethoxyresorufin-O-deethylase activity; EROD) end points, relative liver size (LSI) and condition factor (K) of juvenile female and male rainbow trout exposed to effluents. Unlike previous studies, which have focus either on the specific effects of effluent on fish in laboratory exposures or biotic population statuses downstream of discharge sites, we simultaneously assessed the impacts of PPMES on trout using two approaches: (1) laboratory exposures of tertiary treated PPME produced from processing Eucalyptus globulus or Pinus radiata; and (2) in situ bioassay downstream of the combined discharge of the same pulp mill. Despite an increase in the average gonadosomatic index (GSI) in exposed fish, no statistical differences in gonad size between exposed and unexposed individuals was detected. However, both female and male fish exposed to effluents showed significantly higher concentrations of plasma VTG, so more in fish exposed to Eucalyptus-based effluent when compared to Pinus PPME. In addition, male fish showed intersex characteristics in all exposure assays (Eucaliptus and Pinus) and, despite the low concentration of effluent in the river (<1% [v/v]), similar responses were observed in the caged fish. Finally, EROD activity was induced in both in situ exposures and laboratory assays at the higher PPME concentration (60-85% PPME). This study confirms estrogenic effects in Chilean fish exposed to PPME and the necessity for biological effects monitoring in addition to the assessment of physical-chemical endpoints as required in current government regulations.
Exposure of Reproductively Maturing Rainbow Trout to a New Zealand Pulp and Paper Mill Effluent
Ecotoxicology and Environmental Safety, 2002
Long-term studies on the reproductive 5tness of 5sh under controlled exposure conditions are necessary to address some of the controversy surrounding the 5eld-based studies of pulp and paper e8uent e4ects. This study undertook e8uent exposures of 2# age rainbow trout that were approximately halfway through gonadal growth. Trout were exposed to a mixed thermomechanical/bleached kraft e8uent in 12,000-L 6ow-through exposure tanks at an environmental research facility located at a pulp and paper mill in Kawerau, New Zealand. Trout were exposed to either upstream river water or 10% e8uent in upstream river water and were maintained at a ration of 0.7% of body wet weight during the experiment. Results of the 2-month study indicated that trout survival was not signi5cantly di4erent between e8uent-exposed tanks and reference tanks. There was extensive growth during the exposure but no di4erences were found due to e8uent exposure. Gonadal development was not signi5cantly di4erent between treatments. Steroid hormone concentrations in males and females were not a4ected by e8uent exposure. The e8uent showed no potential to be estrogenic as indicated by a lack of vitellogenin induction in male trout. Other physiological indicators of energy storage and utilization also showed no signi5cant di4erences. Modest induction of hepatic 7-ethoxyresoru5n-O-deethylase (2.5-fold) was the only detectable biological e4ect of the exposure. Biliary concentration of e8uent-related compounds were typical of pulp mill e8uent exposure and further suggested that the source of phytosterols was in fact dietary and not e8uent-derived.
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, 2005
Four pulp and paper mills discharge their effluents in the same section of the Biobio River in central southern Chile. Pulp mill effluents are a very complex mixture with characteristics that depend on the type of raw material, the process technology, and the effluent treatment. To investigate the effect of pulp mill effluent discharges, immature Oncorhynchus mykiss were exposed to river sediments in the laboratory for 29 d. Three sampling areas were defined in a spatial gradient in the river: Preimpact, impact, and postimpact zones relative to the pulp and paper mill discharge areas. Ethoxyresorufin-O-deethylase activities were significantly higher in fish exposed to impact and postimpact sediments when compared to those exposed to preimpact sediments, and higher levels of vitellogenin were observed in the plasma of female fish exposed to impact and postimpact sediments. Histological analysis of the gonadal tissue showed an induction of gonadal maturation in fish exposed to sediment coming from the impact and postimpact zones (oocytes in a vitellogenic state). No site differences were observed in erythrocytes, although differences were noted in the leukocytes in the exposure areas. Finally, the biomarker approach showed evidence that the sediment associated with pulp mill effluent discharges produces some effects in fish under laboratory conditions.