The Relationship between Organic Loading and Effects on Fish Reproduction for Pulp Mill Effluents across Canada.

This study builds upon the work of a multiagency consortium tasked with determining cost-effective solutions for the effects of pulp mill effluents on fish reproduction. A laboratory fathead minnow egg production test and chemical characterization tools were used to benchmark 81 effluents from 20 mills across Canada, representing the major pulping, bleaching, and effluent treatment technologies. For Kraft and mechanical pulp mills, effluents containing less than 20 mg/L BOD5 were found to have the greatest probability of having no effects. Organic loading, expressed as the total detected solvent-extractable components by gas chromatography/mass spectrometry (GC/MS), also correlated with decreased egg laying. Exceptions were found for specific Kraft, mechanical, and sulfite mills, suggesting yet unidentified causative agents are involved. Recycled fiber mill effluents, tested for the first time, were found to have little potential for reproductive effects despite large variations in BOD5 and GC/MS profiles. Effluent treatment systems across all production types were generally efficient, achieving a combined 82-98% BOD5 removal. Further reductions of final effluent organic loadings toward the target of less than 20 mg/L are recommended and can be realized through biotreatment optimization, the reduction of organic losses associated with production upsets and selecting best available technologies that reduce organic loadings to biotreatment.

A survey of Canadian mechanical pulp and paper mill effluents: insights concerning the potential to affect fish reproduction.

Building on breakthroughs recently made at kraft mills, a survey of mechanical pulp and paper mill effluents was undertaken to gain insights concerning potential effects on fish reproduction. Effluents from seven Canadian mills were characterized chemically for conventional parameters such as biochemical oxygen demand (BOD) and total suspended solids (TSS). Each sample was further subjected to solvent extraction followed by gas chromatographic separation for the determination of resin/fatty acids and for the estimation of a gas chromatography (GC) profile index. Each mill effluent was assessed for the potential to affect fish reproduction in the laboratory using a five day adult fathead minnow (Pimephales promelas) egg production bioassay with exposures to 100% effluent. The seven effluents were found to have substantial variation both in terms of chemical characterization and effects on fish reproduction. Temporal variations were also noted in effluent quality at mills sampled on different occasions. Similar to what has been observed for kraft mills, a general trend of greater reductions in egg production caused by effluents with greater BOD concentrations and GC profile indices was noted. Effluents with BOD > 25 mg/L and GC Profile indices >5.0 caused a complete cessation of egg production. At the same time, about half of the total effluents sampled had BOD < 25 mg/L and GC profile indices <5.0 and caused no significant effects on egg production, suggesting these values may be useful as effluent quality targets for mechanical pulp and paper mills. However, 3 out of 14 effluents sampled had BOD < 25 mg/L and GC profile indices <5.0 and caused significant reductions in egg production. The reason(s) for reproductive effects caused by such effluents is presently unclear. The effluent quality parameters considered in this study may require further refinement to address their utility in predicting the adverse reproductive effects induced by effluents from mechanical pulp and paper mills.

Effluent monitoring at a bleached kraft mill: directions for best management practices for eliminating effects on fish reproduction.

A long-term monitoring study was conducted on effluents from a bleached kraft pulp and paper mill located in Eastern Canada. The study was designed to gain insights into temporal effluent variability with respect to fish reproduction as it related to production upsets, mill restarts and conditions affecting biological treatment performance. Final effluent quality was monitored between February 2007 and May 2009 using biochemical and chemical oxygen demand, total suspended solids, resin and fatty acids, a gas chromatographic profiling index, and the presence of methyl substituted 2-cyclopentenones. Selected effluent samples were evaluated for effects on fish reproduction (egg production) using a shortened version of the adult fathead minnow reproductive test. The events relating to negative effects on fish reproduction were upsets of the pulping liquor recovery system resulting in black liquor losses, operational upsets of the hardwood line resulting in the loss of oxygen delignification filtrates, and conditions that reduced the performance of biological treatment (e.g., mill shutdown and low ambient temperatures). The reductions in egg production observed in fathead minnow were associated with biochemical oxygen demand values > 20 mg/L, GC profiling indices > 1.2 and the presence of methyl-substituted 2-cyclopentenones at concentrations > 100 µg/L. This study demonstrated the importance of both in-plant measures for controlling the loss of organics as well as the optimum operation of biological effluent treatment for eliminating effluent-related effects on fish reproduction (egg production) in the laboratory.

Altered reproduction in fish exposed to pulp and paper mill effluents: roles of individual compounds and mill operating conditions.

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.

Use of paired fathead minnow (Pimephales promelas) reproductive test. Part 2: source identification of biological effects at a bleached kraft pulp mill.

Reproductive effects of pulp mill effluents on fish continue to be reported in Canadian waters. Spawning delays, reduced gonad size, and altered levels of sex steroid hormones have been found in both sexes of various fish species exposed to effluents. We initiated a project to identify the source/cause of such effects. In part 1 of this two-part series, we exposed mature adult fathead minnow (FHM; Pimephales promelas) for 21 d to final treated effluent (1% and 100% v/v) from a bleached kraft pulp mill in Terrace Bay (ON, Canada). Results suggested pulp mill effluent from this mill affected reproductive indicators in FHM and effects were dependent on effluent concentration, duration of exposure, and method of data analysis. The main objective of this paper was to use the FHM assay to identify waste stream sources within the mill that affect reproductive indicators. Various process streams were selected, characterized with respect to effluent chemistry and acute toxicity, and a subset was tested on-site with the 21-d FHM bioassay. Results showed that both the combined mill effluent (before secondary treatment) and the combined alkaline stream (CALK) caused both decreased spawning events (approximately 55% for both streams) and decreased egg production (28 and 74%, respectively), and the CALK stream resulted in significant male ovipositor development. By comparing response patterns we were able to identify the CALK stream as a source of compounds at this mill affecting reproductive indicators in FHM and highlight this stream for further toxicity identification evaluation.

Use of paired fathead minnow (Pimephales promelas) reproductive test. Part 1: assessing biological effects of final bleached kraft pulp mill effluent using a mobile bioassay trailer system.

Reproductive effects have been recorded in wild fish in waters receiving pulp mill effluent (PME) since the mid to late 1980s. Laboratory assays with fathead minnow (FHM; Pimephales promelas) have been developed to better understand fish responses to PME. However, observations from laboratory studies have been variable, making it difficult to establish cause/effect relationships. A lack of environmental relevance in these laboratory studies may have contributed to the variability observed. The objectives of the present study were, first, to determine the effects of bleached kraft PME (BKME) on FHM under environmentally realistic conditions (i.e., ambient water and effluent quality) and, second, to determine the suitability of pair-breeding FHM to better link BKME-induced changes in indicators at the biochemical, individual, and population levels. A mobile bioassay trailer was situated on-site at a bleached kraft mill for 60 d, allowing supply of both ambient water (Lake Superior, Canada) and final BKME. The reproductive output of FHM was initially assessed for 21 d to obtain baseline data pre-exposure. At the end of the pre-exposure period, selected breeding pairs were exposed to final BKME (100% v/v and 1% v/v) for 21 d. Results demonstrated a stimulatory response pattern at 1% BKME (e.g., increased egg production) compared to control. In the 100% treatment, spawning events were reduced and fewer eggs were produced during the first two weeks of exposure. Exposure to 100% (v/v) BKME also resulted in ovipositor development in males and development of male secondary sex characteristics in females. Obtaining pre-exposure data and use of pair-breeding FHM in this assay gave a sensitive indication of effluent effects and allowed accurate comparisons of endpoints to be made.

Kraft mill effluent survey: progress toward best management practices for reducing effects on fish reproduction.

Pulp and paper mill effluents have been linked to effects on fish reproduction for more than 25 years. To date, the causes of these effects and remedial strategies have eluded investigators. Recent work has shown that the degree of reproductive effect caused by a mill effluent is related to the overall organic content. If verified, this could lead to breakthroughs for best management practices (BMPs). For this study, the effluents from seven kraft mills were assessed for their ability to reduce egg production in fathead minnow (Pimephales promelas) in the laboratory. At the same time, the effluents were analyzed for three parameters thought to be good indicators of organic losses: biochemical oxygen demand (BOD), methyl-substituted 2-cyclopentenones (cellulose degradation products), and a gas chromatographic (GC) profile index, which integrates the total area of the chromatographic peaks of solvent-extracted effluents using low-resolution mass spectrometry. The results showed that the degree to which the effluents reduced egg production increased with increased organic losses as characterized by BOD and the GC profile index. Therefore, these parameters could be used to guide BMPs at kraft mills according to specific targets: BOD < 20 mg/L and GC profile index equivalent to effluent with BOD of 20 mg/L. Such targets should be achievable by good in-plant control of organic losses and optimized effluent biotreatment systems.

Gene expression profiling of the fathead minnow (Pimephales promelas) neuroendocrine brain in response to pulp and paper mill effluents.

The toxicity of pulp and paper mill effluents (PPMEs) has been greatly decreased, yet some continue to negatively affect fish reproduction. We hypothesized that PPMEs are affecting the brain resulting in decreased reproductive performance. Our goal was to use gene expression profiling to test the hypothesis that PPMEs are having an effect on neural systems in the fathead minnow (FHM; Pimephales promelas) in vivo. Sexually mature male and female FHM were exposed to 100% final biotreated PPMEs from 5 different sources for 5 days. Using an oligo-array (15K genes) we examined the effect of PPMEs on gene expression in the hypothalamus of female fish. We validated selected genes (cholecystokinin, RevErbbeta2, and urotensin I) that were identified by microarray analysis using real-time reverse-transcriptase polymerase chain reaction (RT-PCR). We compared the FHM microarray dataset to multiple microarray datasets from experiments conducted with goldfish injected with different dopaminergic pharmaceuticals to examine whether PPMEs could be affecting the dopamine system. Exposure of FHM to PPMEs resulted in varying degrees of spawning inhibition. Microarray analysis revealed surprisingly few genes in the brain that were commonly affected by the different PPMEs. Real-time RT-PCR confirmed the changes in expression for cholecystokinin, RevErbbeta2, and urotensin I. Comparison of the FHM and goldfish microarray datasets suggest that some PPMEs may be acting on the dopamine system. We show that PPMEs are neuroactive in fish and may be acting through some of the pathways in a manner similar to dopamine.

A long-term, multitrophic level study to assess pulp and paper mill effluent effects on aquatic communities in four US receiving waters: background and status.

An industry-funded, long-term, receiving water study was initiated in 1998/1999 to adress questions about the potential effects of pulp and paper mill effluent discharges on US receiving waters. Although the study continues, the knowledge gained to date provides an opportunity to reflect on the study development process, its progress, and its outcomes. As a backdrop to a series of articles in this special issue describing study results, this article describes the process by which study information objectives were identified as well as the process by which the experimental design was developed. A review of past literature and research identified gaps in long-term population/community data about effluent effects and that, consequently, emerged as a primary information objective. The selected streams for study included 1) Codorus Creek (Pennsylvania, USA), 2) Leaf River (Mississippi, USA), 3) McKenzie River (Oregon, USA), and 4) Willamette River (Oregon) represent a blend of mill process types, coldwater and warmwater stream types, and a range of effluent concentrations. Measurements included numbers of periphyton, macroinvertebrate, and fish communities; the assessment of water and effluent quality; laboratory bioassays; and fish full-life-cycle assays. Information objectives included addressing natural variability and, consequently, the study included long-term temporal (>10 y) and watershed-scale spatial frameworks. Regional-scale ecological risk assessments were performed for each site that aided in placing each site in an ecological and regulatory context. An adaptive-management process is described that allowed for modifications over time as a result of lessons learned as the study progressed. Results from the initial 7 to 8 y of monitoring, as described in the series of articles in this special issue, provide a unique data set with respect to addressing point-source pulp and paper mill effluent discharge concerns and may serve as a template for others to use in developing monitoring or management programs to assess or address water quality conditions or concerns.