To assess anthropogenic impacts of point-source and diffuse discharges on fish populations of the Waikato River, compare responses to different discharges and identify potential sentinel fish species, we sampled wild populations of brown bullhead catfish (Ameiurus nebulosus, (LeSueur, 1819)), shortfin eel (Anguilla australis Richardson, 1848), and common bully (Gobiomorphus cotidianus McDowall, 1975) in the Waikato River. Sites upstream and downstream of: geothermal; bleached kraft mill effluent (BKME); sewage and thermal point-source discharges were sampled. At each site, the population parameters, relative abundance, age structure and individual indices such as: condition factor; and organ (gonad, liver, and spleen) somatic weight ratios; and number and size of follicles per female were assessed. Indicators of fish residence and in some cases exposure to contaminants in discharges were analyzed. Bile chemistry of brown bullhead and shortfin eel was assayed, liver and muscle metal levels were analyzed for brown bullhead and shortfin eel respectively, and stable isotopes of C and N in common bully were measured. Bile, metal and isotopic signatures gave strong evidence that fish had been resident at sites for some time before sampling. Signatures of bile and metal contaminants showed contamination was localised to discharge areas. Gradients in stable isotopes in common bully showed evidence of changes in water sources and anthropogenic effects along the river. Biochemical variables, hepatic ethoxyresorufin-O-deethylase (EROD) and plasma steroids indicated exposure and response of brown bullhead and shortfin eel to pulp and paper contaminants at the BKME site. Physiological (blood) variables showed fish largely responded in a predictable way to elevated water temperatures at discharge sites at time of sampling, however total haemoglobin of brown bullhead and common bully blood failed to increase at the BKME site despite elevated temperatures and low dissolved oxygen. Growth rates, condition factor, age structure, and gonadosomatic index (GSI) suggest that discharges with significant heat or nutrients benefit brown bullhead despite physiological impairment at the BKME site. Shortfin eel individuals also benefited from heated water discharges. No consistent impacts on common bully health were obvious at individual discharge sites, or cumulatively along the river due to the gradual deterioration in water quality downstream. Common bully individuals also benefited from heat in discharges but lack of juveniles at sites where numerous juvenile brown bullhead were found, suggest that unlike brown bullhead populations, common bully populations were not responding with significant recruitment. Although I found little evidence of toxic effects of discharges on shortfin eel, caution is required in assessing the potential of contaminants to impact eel populations due to the life history of shortfin eel, and exploited nature of populations. For example, reproductive damage suffered by adult eels may not immediately manifest itself in the effected population due to temporal delays in gonadal maturation, and recruitment, and single panmictic populations supplementing recruitment of impacted populations. Distinct changes in population parameters at each of the paired sites and changes in individual variables showed that fish responded to discharges. The range of responses in species suggests different sensitivity to contaminants and amount of benefit which each species receives from heat in discharges. In these terms shortfin eel would be the most resistant, then brown bullhead and lastly common bully. Interpretation of population-level impacts at the geothermal and BKME discharge sites is made difficult due to benefits of additional heat. There is also the possibility that detection of sub-lethal or chronic effects on sensitive juvenile life-stages may be being hidden by compensatory density population responses. Responses and life history of common bully made them the preferred indicator species of the three species sampled, and supported overseas examples using small-bodied fish species as sentinels.

Previous research identified distinct genetic, life-history and reproductive differences between populations of common bully (Gobiomorphus cotidianus) upstream and downstream of a pulp and paper mill outfall on the Tarawera River in the Bay of Plenty, New Zealand. This thesis sought to investigate the distribution of amphidromous and non-amphidromous common bully in the Tarawera River by examining fish collected from upstream (37 km inland) and downstream (20 km inland) locations and comparing them to fish from similar inland locations (40 km and 17 km inland, respectively) in the nearby Rangitaiki River. Otolith microchemistry revealed life-history differences between upstream and downstream populations and stable isotope analysis ensured long-term site residency. Amphidromy dominated in the downstream river populations, while the disappearance of diadromous fish generally occurred with inland distance. A mixture of diadromous and non-diadromous fish were found in the upstream Rangitaiki, while a complete absence of diadromous recruits was found in the upstream Tarawera River. A reduction in oculoscapular canal structures also coincided with loss of diadromy in fish from both rivers. Temporal reproductive divergence was investigated through track annual trends in gonadosomatic index. The Tarawera River receives significant inputs from numerous industrial, municipal and natural sources, most notably from two pulp and paper mills. In the absence physical barriers in the Tarawera, it has been hypothesised that the lack of diadromous recruits in the upstream Tarawera River may be related to aquatic discharges in the downstream river. A behavioural study was performed to examine the hypothesis that pulp and paper mill effluent may be acting as a chemical barrier to fish migration within the river. A dual-choice chamber was employed to examine the responses of common bully exposed to a range of effluent concentrations (100, 50, 25, 12.5, 0% v/v). Fish exhibited significant avoidance responses when exposed to 100 and 50% effluent concentration, while no avoidance was observed at effluent concentrations below 50% This study demonstrated that common bully show a strong preference for river water when simultaneously exposed to effluent, albeit at environmentally unrealistic concentrations (i.e. greater than 15%), implicating potential for this effluent to act as a chemical barrier in the Tarawera River. Following the establishment of reproductive timing of common bully in the Tarawera River, a wild fish health assessment was undertaken to investigate the effects of long-term effluent exposure in situ. Adult common bully were sampled downstream of the mill influence and compared to an appropriate reference population from the downstream Rangitaiki River. Male and female fish from the Tarawera River demonstrated 6- to 9-fold greater ethoxyresorufin-O¬-deethylase (EROD) activity compared to reference fish, indicating exposure to organic contaminants in this river. Tarawera females showed some minor variation in hematological variables including decreased mean cell volume (MCV), mean cell haemoglobin (MCH) and increased total white blood cell count (WBCC) suggestive of an immune response. Slightly greater ovarian follicular steroid production in Tarawera fish potentially indicates some form of endocrine alteration. However, this response may also be related to differences in reproductive synchrony and gonadal development between the two fish populations.

Stable isotope probing (SIP) is an established technique that can be applied to identify the metabolically active micro-organisms within a microbial population. The SIP method utilises an isotopically-labelled substrate and PCR techniques to discern the members of a microbial community that incorporate the isotope into their DNA or RNA. The current literature gap around using 15N isotopes with RNA-SIP offers real potential and advantages for targeting and identifying active members from mixed communities involved in global biogeochemical nitrogen cycling. This study specifically investigated whether nitrogen based compounds can be used as substrates in RNA-SIP methodologies and whether they can in turn be used to probe mixed community environments known to be actively fixing nitrogen. The nitrogen-limited systems targeted represented an ideal opportunity to assess the suitability of 15N-RNA-SIP approaches due to their known high nitrogen fixation rates. Identifying these nitrogen-fixing bacteria could provide a better representation analysis of the community, leading to an improved prediction on how to manage and optimise the treatment performance of target waste systems and to exploit the unique bioconversion properties of these types of organisms. Initially, the project undertook methodological proof of concept by using a soluble nitrogen source, 15NH4Cl, to label the RNA of Novosphingobium nitrogenifigens and a mixed microbial community. Successful separation of the 14N- (control) and 15N-RNA was achieved for both pure and mixed communities using isopycnic caesium trifluoroacetate (CsTFA) gradients in an ultracentrifuge. The usefulness of this technique to identify active diazotrophs in real environmental samples was tested using a nitrogen-fixing community from a pulp and paper wastewater treatment system. After growing the mixed culture with 15N2 as the sole nitrogen source, the labelled RNA was extracted and fractionated using isopycnic centrifugation in CsTFA gradients. The community composition of the active nitrogen-fixing community in the 15N2 enriched fraction was analysed by establishing a 16S rRNA gene clone library containing over 200 members. These were analysed by comparison with published sequences and by phylogenetic analysis. It was found that the more isotopic label substrate incorporated, the further the buoyant density (BD) separation between 15N- and 14N-RNA. Novosphingobium nitrogenifigens gave an average BD shift of 0.03 + 0.004 g ml-1 (95.0 atom % 15N) with 15NH4Cl. For mixed communities the average BD shift was 0.02 + 0.004 g ml-1 (80.0 atom % 15N) with 15NH4Cl and 0.013 + 0.002 g ml-1 (32.6 atom % 15N) when using 15N2. Clone library analysis of 16S rRNA genes present in the enriched 15N-RNA fraction of the mixed community was shown to consist of a diverse population of bacteria as indicated by a Shannon Weaver index value of gt;2.8. Three dominant genera (Aeromonas, Pseudomonas and Bacillus) were identified by comparison with published sequences and phylogenetic analysis. Many other groups not known as archetypal nitrogen-fixing bacteria were also identified, demonstrating that 15N2-RNA-SIP provides a useful tool for the identification of important and previously unknown contributors to nitrogen fixation in a range of environments. Overall, this project has established that nitrogen based RNA-SIP is a powerful tool that can be used successfully and reproducibly with both pure and complex mixed microbial communities to study active diazotrophs in environmental samples.

<p>This work explores the usefulness of fish bile analysis in combination with biomarkers for identifying and evaluating new environmental contaminants in the aquatic environment. It illustrates how bile analysis can be used together with biomarkers to assess the causes of estrogenic effects, to identify chemicals in the aquatic environment that are taken up by fish, and to monitor environmental exposure.</p><p>In a first application, fish exposed to sewage treatment plant effluent were studied. Elevated levels of vitellogenin in the exposed fish demonstrated that estrogenic effects occurred. Several estrogen disrupting substances were identified in the fish bile, and analysis of water samples confirmed that these substances were present in the effluent. The synthetic estrogen 17a-ethinylestradiol, which is known to be present in sewage treatment plant effluent, was shown for the first time to be taken up by fish. Considering the reported potencies of the detected substances, it was concluded that 17a-ethinylestradiol was the major contributor to the estrogenic effects.</p><p>Chemical analysis of bile was used to identify rubber additives that were released from tires immersed in water. The bile of rainbow trout held in the water contained high levels of metabolites of PAHs and aromatic nitrogen compounds. Several biomarkers were also measured in the exposed fish, and EROD induction and oxidative stress were observed. Based on the bile analysis observations together with knowledge of toxicological mechanisms, it was postulated that the EROD induction was due to the PAHs, while aromatic nitrogen compounds caused the oxidative stress.</p><p>Resin acids in fish bile proved to be a good indicator of exposure in a chronic long-term study of rainbow trout exposed to effluent from a total chlorine free (TCF) pulp mill. Elevated levels of GST (gluthatione-S-transferase) and GR (gluthatione reductase) activity, and the presence of DNA adducts after a two month recovery period, indicated that compounds in the pulp mill effluents have persistent effects. In addition to characterising the exposure of the fish to the effluent, the analysis of the resin acids in the bile provided evidence of accidents in the pulp mill that the existing process monitoring system had not detected.</p><p>Resin acids in bile were also found to be a valuable indicator of exposure to pulp mill effluents for eelpout living in the Baltic Sea. A correlation between resin acid levels in bile and skewed sex ratios provided an important link in the chain of evidence that substances in the pulp mill effluents cause male bias of the eelpout embryos.</p><p>A particularly good example of the potential of bile analysis was the identification of a previously unknown environmental contaminant. A large peak was observed in the bile extracts of fish that had been exposed to sewage treatment plant effluent. This peak was identified as triclosan, which demonstrated its presence in sewage treatment plant effluent. Other work went on to show that it is a common contaminant of the aquatic environment. The ability of fish to concentrate contaminant metabolites in bile to levels very much higher than in the environment, and the comparatively low levels of analytic interferences, make bile a particularly attractive matrix to search for new, unknown organic pollutants</p>

The pulp and paper industry has been struggling to meet the new and stringent TRS (Total Reduced Sulphur) emission compliance standards established in recent years. However, a new approach by some regulatory bodies gives intricate operational parameters a new and important role in achieving environmental compliance. TRS compounds that cause the distinctive pulp mill odour, originate from sodium sulphide in white liquor used in the kraft pulping process. Up to 20% of TRS emissions could originate from the lime kiln and lime mud solids content is one of the operational parameters that could help reduce the TRS emissions from the lime kiln. Residual sodium sulphide in the lime mud that results in TRS gases, is dissolved in the moisture content of the mud. Although efficient lime mud washing can remove most of the residual sodium sulphide, the remaining moisture content of the mud still contains some sodium sulphide. Therefore, improved lime mud dewatering can be effective in reducing the TRS emissions from the lime kiln. Data presented in this study confirms that as the lime mud solids content increases, TRS emissions from the lime kiln decrease. Data analysis demonstrates a negative linear correlation at 5% significance level between TRS emissions and lime mud solids.

The pulp and paper industry has been struggling to meet the new and stringent TRS (Total Reduced Sulphur) emission compliance standards established in recent years. However, a new approach by some regulatory bodies gives intricate operational parameters a new and important role in achieving environmental compliance. TRS compounds that cause the distinctive pulp mill odour, originate from sodium sulphide in white liquor used in the kraft pulping process. Up to 20% of TRS emissions could originate from the lime kiln and lime mud solids content is one of the operational parameters that could help reduce the TRS emissions from the lime kiln. Residual sodium sulphide in the lime mud that results in TRS gases, is dissolved in the moisture content of the mud. Although efficient lime mud washing can remove most of the residual sodium sulphide, the remaining moisture content of the mud still contains some sodium sulphide. Therefore, improved lime mud dewatering can be effective in reducing the TRS emissions from the lime kiln. Data presented in this study confirms that as the lime mud solids content increases, TRS emissions from the lime kiln decrease. Data analysis demonstrates a negative linear correlation at 5% significance level between TRS emissions and lime mud solids.

The overall objective of the research conducted and described in this thesis was to develop an environmentally relevant bioassay to assess the effects of complex effluents on a sentinel fish species. A short-term fathead minnow (FHM) reproductive bioassay was utilized to assess the effects of industrial effluents on multiple levels of biological organization (sub-organismal to population endpoints). The FHM bioassay was tested in both lab and on-site investigations using an artificial stream system. The incorporation of trophic-transfer into the bioassay was also developed to quantify the importance of contaminated food as a source of exposure. This work was conducted in two key phases. Phase I focused on testing and developing the FHM bioassay, in the lab and on-site with pulp mill effluent (PME), to firstly document response patterns and, secondly, to conduct an investigation of cause study. Phase II focused on developing the trophic-transfer system to document responses to metal mine effluent (MME) in the lab and on-site in an artificial stream system. Development of the trophic-transfer system was also conducted during this phase to compare responses to standard water-only exposures. <p>In Phase I, exposure to PME in both the lab and field studies resulted in disruptions in egg production and spawning events. By focusing on identifying response patterns I was able to determine that the effects observed were indicative of an estrogenic response. I was also able to identify a process stream that was the potential cause of responses observed after exposure to final effluent. Isolation of this process stream will assist the mill in developing approaches for future mitigation. The results from this research will also provide additional data for the environmental effects monitoring (EEM) program for pulp and paper and investigation of cause studies on a national basis. <p> In Phase II, in both the field and laboratory investigations, significant decreases in reproductive output (egg production and spawning events) were observed in the water-only system exposures. Significant decreases in hatching success and increases in deformities were observed in the trophic-transfer system only, suggesting that the combination of both food and water was important in assessing the effects on the F1 generation. Overall, the responses in the trophic-transfer system were not comparable between the lab and field studies. In the lab study, significant decreases in reproductive output occurred, compared to the field study where significant increases in egg production and spawning events occurred. In addition, the effects on the F1 generation in the field study were not as severe as those observed in the lab investigation. It was concluded that the presence of reference water and the environment within the trophic-transfer system were responsible for this reduction in toxicity. <p> Phases I and II of this research have made significant contributions to artificial stream development within Canada for the assessment of industrial effluents and their effects on aquatic biota. The results from these studies have also demonstrated that environmentally relevant testing is essential if we are to accurately assess effects on aquatic biota. Future development and application of this bioassay should be towards developing a standardized approach for not only assessing the effects of industrial effluents in a comparative manner, but also in investigation of cause studies.

This research project originated from interest in assessing the potential of enzyme technology (particularly laccase-based systems) for the biomodification of sisal specialty fibres by using environmentally friendly processes. This doctoral work focused on two different research lines, namely: biobleaching and enzymatic functionalization of sisal pulp fibres. The study was started by assessing the use of natural, potentially cost-effective phenolic compounds as substitutes for expensive, potentially toxic laccase mediators. The tendency of natural phenols to either promote delignification or couple onto pulp was examined with a view to assessing their potential for either bleaching or functionalizing sisal fibres. In the biobleaching study, totally chlorine free (TCF) sequences were implemented in order to compare the efficiency of a selected natural mediator and a well-known synthetic mediator, both in the presence and absence of a xylanase pre-treatment. The effluents resulting from each stage in the sequence were analysed with a view to assessing the environmental impact of the laccase treatments ¿a scarcely explored aspect of biobleaching sequences. The xylanase stage proved highly efficient in reducing the HexA content of sisal fibres and in boosting the bleaching effect of the laccase treatments. The proposed TCF sequences provided high-cellulose sisal pulp with brightness above 80% ISO and a reduced HexA content; also, they exhibited improved performance and a reduced impact on effluent properties relative to the use of the synthetic mediator. Two different approaches to fibre functionalization were explored, namely: lignin modification (biografting) and cellulose modification (laccase¿TEMPO oxidation). Biografting of phenolic compounds was for the first time studied in sisal pulp. Covalent binding of the originally assayed phenolic compounds to sisal fibres during the laccase treatment was exposed by a novel analytical approach based on pyrolysis-GC/MS. The phenolic compound showing the highest tendency to couple to fibres was selected to investigate biografting under different reaction conditions and to evaluate the extent of phenol coupling via various pulp properties. Biografting efficiency was enhanced by refining the fibres prior to the enzyme treatment, which provided improved strength-related properties in the resulting paper. The use of the laccase-TEMPO system to oxidatively modify cellulose and improve strength-related properties in sisal pulp was for the first time evaluated as an environmentally friendly alternative to existing halide-based systems. The first part of this study revealed that the laccase¿TEMPO system considerably improved wet strength in sisal pulp by effect of the formation of a substantial amount of aldehyde groups in cellulose chains that facilitated inter-fibre bonding through hemiacetal linkages. The influence of process variables on various properties of the oxidized fibres and resulting paper was assessed by using a three-variable statistical plan. The conditions maximizing functionalization and the improvement in paper strength properties were used to design treatments of increased efficiency that exposed the potential of laccase¿TEMPO oxidation for biorefining pulp fibres. Analytical methods including pyrolysis-GC/MS, polyelectrolyte titration, conductimetric titration, carbohydrate determination by HPLC, fibre morphology analysis by SEM and thermogravimetry were used to both characterize the raw material and gain a better understanding of the reaction mechanisms behind the different laccase-based treatments. Some of the analyses were performed by collaborating research groups at IRNAS (Seville, Spain) and the Department of Chemical Engineering of the University of Huelva (Spain). Also, part of this doctoral work was conducted at the Institute of Paper Science and Technology of the Georgia Institute of Technology (Atlanta, USA).

Freshwater availability, allocation and quality are increasingly becoming a concern in Canada. Canada’s apparent availability of abundant freshwater is not what it was thought to be. With increasing competition for water sources and inadequate management systems water use and management approaches are being re-examined. While urban and municipal water uses are highly studied, Canadian industrial water use is not. Despite that industrial water use is understudied, the Canadian pulp and paper industry is a major user of water and contributes to quality water issues where mills are located. This thesis is divided into two equally important parts. First, this research seeks to understand the influences and prevailing conditions on the decision-making framework of pulp and paper mills. Second, it seeks to understand how the prevailing conditions affect the applicability of the water soft-path concept in the pulp and paper industry. It will contribute to the literature of Canadian industrial water management. This research specifically examines the applicability of the water soft-path concept under the stresses and realities of the systems of influence identified as market forces, policy and regulation, and technology faced by the Canadian pulp and paper industry. Corporate culture was an implicitly common thread that ran through these systems of influence. A variety of methods were used in this study including, a literature review conducted by themes, surveys, interviews, analysis of archival data and backcasting were used as the methodological approaches. The literature review was conducted by themes of water management, technology, market forces, regulation and corporate culture. Surveys were conducted to gain water use data from specific mills but a low response rate required a widening of the research boundaries. Interviews were conducted with government officials, industry representatives, and environmental non-governmental organizations. The interviews contributed to the boundary setting and understanding of the influences that impact decision-making for industry. The analysis of archival data was to better understand how water use in pulp and paper mills has changed through the years. Understandably the systems of influence (market forces, policy and regulation, and technology) work independently and together to create a complex environment in which decisions on water use in pulp and paper mills are made. The complexity of the decision-making framework is great and the barriers to water soft-path application difficult. Market forces are less capable of addressing environmental externalities such as water. Regulation and policy has yet to address water use in industry. Technology does provide an important opportunity for efficient water use and application of the pulp and paper industry. Ultimately, the Canadian pulp and paper industry is in a redefining moment where opportunity exists to create a new direction and approach to water use in the Canadian pulp and paper industry.

Hallsta paper mill uses large amounts of water during the pulp and paper making processes. The wastewater is treated in two separate activated sludge processes referred to as BIO 1 and BIO 2. The main aim is to reduce the organic substance content, measured as COD (Chemical Oxygen Demand) before the wastewater is released into the environment. Since the effluent produced is very rich in organic substances, the addition of nitrogen (N) and phosphorus (P) are necessary to give satisfactory treatment results. Problems with sludge bulking sometimes occur which lead to increased levels of suspended matter and nutrients in the effluent. Periods of high concentration of phosphorus is also of great concern. The aim of this study is to investigate the causality behind the sludge bulking, with the help of scientific literature and analysis of previously compiled data using multivariate data analysis. COD and nutrients present in the influent will also be surveyed to identify variations in concentration caused by the paper making process. The scientific literature points toward low nitrogen and phosphorus levels as the major causes of the bulking sludge, as this leads to increased growth of filamentous bacteria to the detriment of the preferred flocculating species. A full scale experiment was conducted in an attempt to evaluate sufficient dosage levels of N and P to reduce this problem. The experiment has produced stabile results with low percentages of suspended organic matter and nutrients. Evaluation of the previously collected data using Multivariate Data Analysis did not lead to any correlation between variables being found, other than the impact of sludge concentration on COD-reduction. No relevant explanation to periods of high phosphorus levels could be found neither in the scientific literature nor in the multivariate data analysis. Bleaching of the paper pulp, with hydrogen peroxide and lye, results in a subsequent increase in COD concentration in the wastewater. The study shows an increased COD in the water approximately 12 hours after the increased dosing of these bleaching agents. Also the amounts of easily degradable COD increases following increased dosage of hydrogen peroxide and lye. The content of easily accessible phosphorus in the influent represents a minimum of 25 % and 20 % of the amount theoretically required for BIO1 and BIO2 respectively. The influent levels of nitrogen represents 33 % and 100 % for BIO1 and BIO2 respectively. From this study it has been possible to deduce that a controlled dosage of nutrients in relation to the incoming level of COD, is of great importance to the efficient treatment of COD rich effluent in the systems under investigation at Hallsta Papermill. / Vid Hallsta Pappersbruk används stora mängder vatten vid tillverkningen av massa och papper. Det förorenade processvattnet renas i två separata anläggningar, BIO1 och BIO2, båda med biologisk rening av typen aktivt slam. Avloppsvattnets höga halter av kolföreningar, vilka analyseras som COD (Chemical Oxygen Demand), kräver extra tillsats av kväve och fosfor för att erhålla tillfredsställande reningsresultat. I den biologiska reningen uppstår periodvis problem med slamsvällning, något som resulterar i höga utsläpp av suspenderat material och organiskt bundna näringsämnen. Även perioder med höga fosforutsläpp förekommer. Syftet med examensarbetet är att genom litteraturstudier och analys av historiska data, med hjälp av multivariat dataanalys, utreda orsakerna till problemen. Även en kartläggning av variationerna i inkommande avloppsvatten med avseende på COD och lättillgängliga näringsämnen genomförs. Utifrån information från litteraturen tyder problemen med slamsvällning ofta på bristande tillgång på kväve och fosfor vilket bland annat kan resultera i kraftig tillväxt av filamentösa bakterier. För att komma till rätta med problemen startades ett styrt försök med beräknade optimala doseringskvoter. Försöket resulterade i stabil drift för båda anläggningarna med låga utsläppsnivåer av både suspenderat material och näringsämnen. Resultatet från utvärderingen av driftsdata med multivariat analys visar att slamhalten har betydelse för COD-reduktionen i BIO1. Någon förklaring till problemen med höga fosforutsläpp har inte framkommit vare sig ur litteraturstudie eller multivariat dataanalys. Vid alkalisk oxiderande blekning av termomekanisk pappersmassa ökar den totala COD-koncentrationen i avloppsvattnet i samband med höjd dosering av blekkemikalier. Studien visar att en ökad COD-koncentration kan observeras i vattnet in till BIO1 ca 12 timmar efter förhöjd dosering av blekkemikalier. Samtidigt ökar andelen lättnedbrytbart COD till följd av ökad blekningsgrad. Kartläggningen av inkommande vatten visar att mängden lättillgängligt kväve i avloppsvattnet in till biologierna motsvarar ca 33 % och 100 % av det teoretiskt beräknade behovet för BIO1 respektive BIO2. För fosfor uppgår motsvarande till minst 25 % respektive 20 % för BIO1 och BIO2. Under examensarbetets gång har det kunnat konstateras att dosering av närsalter i förhållande till inkommande mängd COD är av stor vikt för en väl fungerande rening av det COD-rika avloppsvattnet vid Hallsta Pappersbruk.