The combination of moving bed biofilm reactors and membrane bioreactors (MBBR-MR) can compensate for the drawbacks of both of these systems and further increase their acceptance and application in wastewater treatment industries. Despite the potential benefits of a MBBR-MR technology there has only been limited study of this configuration. The present study consisted of an overall assessment of the performance of a combined MBBR-MR system under high and low loading rates. Since colloidal matter in mixed liquor suspended solid (MLSS) is considered as one of the important contributors to membrane fouling, pre-treatment of membrane feed by coagulation was investigated for improving membrane performance. The performance of the MBBR-MR was assessed based on its chemical oxygen demand (COD) removal efficiency and membrane fouling mechanisms. The study was carried out using pilot-scale MBBR and bench-scale batch membrane filtration setups (Millipore Inc. Bedford, MA). The pilot MBBR had a working volume of 1.8 m3 and a 30% carrier fill fraction. The MBBR was operated with loading rates of 160 ± 44 g/m2/d (hydraulic residence time (HRT) of 4.6 h) and 223 g/m2/d (HRT of 2.6 h). The MBBR feed was obtained from a starch recovery line in a potato chip processing factory. The carriers were mixed by coarse bubble aeration and the dissolved oxygen (DO) was maintained above 2 mg/l. Preliminary jar test trials (based on turbidity removal) were performed in order to obtain an optimal dosage of coagulants for subsequent ultrafiltration (UF) tests. The efficiency of three coagulants (alum, ferric chloride and a blend of polyaluminum chloride and polyamine) was evaluated. The membranes were composed of polyethersulfone (PES) and had a pore size of 0.05 microns. The results of this study indicate that the combination of MBBR with membrane filtration can be operated at relatively high loading rates to yield a constant high quality permeate that is suitable for water reuse purposes. Fouling of the membrane by the wastewater was found to be substantially reduced by treatment with the MBBR. The reversible and irreversible fouling of the MBBR effluent were 56 and 63%, respectively, of that observed with the raw wastewater. The MBBR Loading-rate was found to affect treatment efficiency of the MBBR-MR and membrane performance. Operation under the elevated loading-rate conditions HRT = 2.6 hours) resulted in an increase in the irreversible fouling of the membranes (60% on average). The addition of all the coagulants in this study was found to decrease the fouling of the membrane. However, the extent of the pre-coagulation effect on membrane fouling was found to strongly depend on the type and dosage of the coagulant and the MBBR effluent characteristics. All the coagulants were effective in decreasing membrane fouling at their optimal dosages which was determined in preliminary jar tests. Ferric chloride performed the best as a pretreatment coagulant compared to alum (Aluminum sulfate) and the coagulant blend with reductions in both reversible and irreversible fouling (43-86% and 51-71%, respectively) and increased consistency (in decreasing fouling) as compared to the other coagulants. Alum had no effect on irreversible fouling and the coagulant blend significantly increased irreversible fouling in some trials (up to 196% or by a factor of 3 when overdosed). Additionally, alum and the blend were, on average, 29% and 7%, less effective than ferric chloride in reducing reversible fouling under the conditions and dosages tested.

The combination of moving bed biofilm reactors and membrane bioreactors (MBBR-MR) can compensate for the drawbacks of both of these systems and further increase their acceptance and application in wastewater treatment industries. Despite the potential benefits of a MBBR-MR technology there has only been limited study of this configuration. The present study consisted of an overall assessment of the performance of a combined MBBR-MR system under high and low loading rates. Since colloidal matter in mixed liquor suspended solid (MLSS) is considered as one of the important contributors to membrane fouling, pre-treatment of membrane feed by coagulation was investigated for improving membrane performance. The performance of the MBBR-MR was assessed based on its chemical oxygen demand (COD) removal efficiency and membrane fouling mechanisms. The study was carried out using pilot-scale MBBR and bench-scale batch membrane filtration setups (Millipore Inc. Bedford, MA). The pilot MBBR had a working volume of 1.8 m3 and a 30% carrier fill fraction. The MBBR was operated with loading rates of 160 ± 44 g/m2/d (hydraulic residence time (HRT) of 4.6 h) and 223 g/m2/d (HRT of 2.6 h). The MBBR feed was obtained from a starch recovery line in a potato chip processing factory. The carriers were mixed by coarse bubble aeration and the dissolved oxygen (DO) was maintained above 2 mg/l. Preliminary jar test trials (based on turbidity removal) were performed in order to obtain an optimal dosage of coagulants for subsequent ultrafiltration (UF) tests. The efficiency of three coagulants (alum, ferric chloride and a blend of polyaluminum chloride and polyamine) was evaluated. The membranes were composed of polyethersulfone (PES) and had a pore size of 0.05 microns. The results of this study indicate that the combination of MBBR with membrane filtration can be operated at relatively high loading rates to yield a constant high quality permeate that is suitable for water reuse purposes. Fouling of the membrane by the wastewater was found to be substantially reduced by treatment with the MBBR. The reversible and irreversible fouling of the MBBR effluent were 56 and 63%, respectively, of that observed with the raw wastewater. The MBBR Loading-rate was found to affect treatment efficiency of the MBBR-MR and membrane performance. Operation under the elevated loading-rate conditions HRT = 2.6 hours) resulted in an increase in the irreversible fouling of the membranes (60% on average). The addition of all the coagulants in this study was found to decrease the fouling of the membrane. However, the extent of the pre-coagulation effect on membrane fouling was found to strongly depend on the type and dosage of the coagulant and the MBBR effluent characteristics. All the coagulants were effective in decreasing membrane fouling at their optimal dosages which was determined in preliminary jar tests. Ferric chloride performed the best as a pretreatment coagulant compared to alum (Aluminum sulfate) and the coagulant blend with reductions in both reversible and irreversible fouling (43-86% and 51-71%, respectively) and increased consistency (in decreasing fouling) as compared to the other coagulants. Alum had no effect on irreversible fouling and the coagulant blend significantly increased irreversible fouling in some trials (up to 196% or by a factor of 3 when overdosed). Additionally, alum and the blend were, on average, 29% and 7%, less effective than ferric chloride in reducing reversible fouling under the conditions and dosages tested.

There is an urgent need to develop ammonia removal treatment systems for municipal and industrial wastewater treatment due to the increasingly stringent ammonia effluent discharge regulations implemented by Canada, the United States, and the European Union. The objective of this dissertation is to develop new understanding and advance the current design and operation of total ammonia nitrogen (TAN) removal via the moving bed biofilm reactor technology (MBBR) for municipal and industrial wastewaters. The first specific objective is to develop a passive, low operationally intensive, efficient and robust design strategy for municipal wastewater treatment to achieve partial nitritation (PN) as a pre-treatment to anammox treatment without using control strategies such as operating at low dissolved oxygen, or the use of inhibitors. This first objective includes developing new knowledge of the biofilm, biomass and microbiome of attached growth PN systems. The second specific objective is to investigate the impact of defining a maximum biofilm thickness, via bio-carrier design, to enhance the effects of free nitrous acid inhibition for PN of municipal wastewaters. The third objective is to investigate the effect of influent copper concentration on nitrifying MBBR systems over long-term operations, to demonstrate the feasibility of the nitrifying MBBR as a solution for TAN removal from gold mining wastewaters. The results pertaining to the first objective, achieved via a study investigating the operation of a nitrifying moving bed biofilm reactor at elevated TAN surface area loading rates (SALRs) of 3, 4, 5, and 6.5 g TAN/m²∙d with the aim of achieving passive PN, demonstrates that operating at a TAN SALR value of 6.5 g TAN/m²∙d can achieve PN without restricting dissolved oxygen or using inhibitors. Operating at a TAN SALR value of 6.5 g TAN/m²∙d achieves a TAN surface area removal rate (SARR) of 3.5 g TAN/m²∙d, and a nitrite accumulation of 99.8% of the oxidized TAN, demonstrating the suppression of nitrite oxidizing bacteria (NOB) activity, while achieving elevated TAN SARR values. At the molecular-scale, there is a statistically significant change in the ammonia oxidizing bacteria (AOB) to NOB ratio from 1:2.6 to 8.7:1 as the TAN SALR increases from 3 to 6.5 g TAN/m²∙d; however, even at a TAN SALR value of 6.5 g TAN/m²∙d there is an NOB abundance of approximately 2%; thus demonstrating that NOB remain present in the biofilm, while their activity is suppressed by operation at elevated TAN SALR values. Furthermore, this system was shown to achieve stable PN consistently for over a period of 10 months of operation, demonstrating a robust, passive, low operational strategy for attached growth PN. The second objective of this dissertation is addressed through a study that compared the carrier design of defined maximal biofilm thickness (z-prototype carrier) to undefined maximal biofilm thickness (chip-prototype carrier) for PN via free nitrous acid inhibition of tertiary, low carbon, municipal wastewaters. The study demonstrates that defined maximal biofilm thickness is a preferred design choice to achieve attached growth PN. The chip-prototype carrier shows biofilm thicknesses and biofilm mass values that are ten-fold higher than the z-prototype carrier, which is shown to contribute to the impact of free nitrous acid on AOB and NOB activities. The z-prototype carrier shows PN is achieved after 3 hours of exposure to free nitrous acid while the chip-prototype carrier does not achieve PN within this same time of exposure. Therefore, the defined maximal biofilm thickness carrier is identified in this research as the preferred design option to achieve attached growth PN for municipal, low carbon, tertiary wastewater treatment. The results of the third objective, achieved via a study investigating the effects of influent copper concentrations on nitrifying MBBR during long term operations to gold mining wastewaters, demonstrates that there is no AOB inhibition in attached growth systems exposed to 0.1, 0.3, 0.45, and 0.6 mg Cu/L for long exposure times. A trend of increasing nitrite accumulation with increasing influent copper concentrations is shown, indicating that NOB inhibition occurs at influent copper concentrations of 0.3 mg Cu/L and greater, with the greatest NOB inhibition observed with an influent copper concentration of 0.6 mg/L. There is no statistically significant difference in biofilm characteristics at the copper concentrations tested; however, there is a trend of increasing biofilm thickness and biofilm roughness with increasing copper concentrations. This study demonstrates the resilience of the nitrifying biofilm to copper inhibition and demonstrates that the nitrifying MBBR is a promising system for removing TAN in mining wastewater in the presence of copper.

Introdução sistemas de tratamento de esgotos, quando aplicados ao tratamento de lixiviado de aterro sanitário, não têm apresentado bons resultados. Por conta disso, faz-se necessário o desenvolvimento de alternativas para o tratamento de lixiviado. Objetivo avaliar o pós-tratamento de lixiviado de aterro sanitário por wetland construído de fluxo subsuperficial horizontal e reator biológico de leito móvel (RBLM). Materiais e métodos foram realizados dois experimentos com tratamento de lixiviado após remoção de amônia por dessorção. No experimento 1 utilizou-se wetlands construídos de fluxo horizontal em escala de bancada (volume total de 30,8 L), plantados com Cyperus papyrus, Heliconia psittacorum e Gynerium sagittatum, e um controle sem vegetação, preenchidos com pedrisco calcário. O sistema foi alimentado com lixiviado diluído em água em diferentes proporções (entre 10 por cento e 30 por cento ) com concentração média de DQO entre 336 e 750 mg.L-1 e nitrogênio amoniacal (N-NH4) entre 47 e 199 mg.L-1. A operação ocorreu em três etapas: (1ª) alimentação contínua e TDH médio entre 2,7 e 5,3 d; (2ª) regime de ciclos de recirculação do efluente, para avaliar o efeito do aumento de TDH para 21 dias; (3ª) alimentação contínua de duas unidades wetlands em série, visando elevar o TDH (entre 8,1 e 9,9 d) sem recirculação. O experimento 2 foi implantado em escala piloto, com uma unidade de dessorção de amônia seguida de um Reator Biológico de Leito Móvel (RBLM) e um biofiltro anóxico. O RBLM (volume efetivo de 380 L) foi preenchido em 50 por cento com anel pall. Resultados e discussão No experimento 1, obteve-se baixa remoção de DQO com médias inferiores a 40 por cento e concentração final entre 270 e 750 mg.L-1. No entanto, verificou-se remoção de N-NH4, principalmente na 2ª e 3ª etapas, com médias de remoção entre 43 por cento e 81 por cento , resultando em concentração de 20 a 223 mg.L-1, com influência do TDH. No experimento 2, problemas técnicos durante a instalação e operação inicial e interdição do campus da USP-Leste prejudicaram a adaptação do reator e impediram o seu monitoramento por tempo suficiente para geração de dados consistentes, sendo interrompido na fase inicial. Conclusões A baixa eficiência para DQO provavelmente está relacionada à recalcitrância do lixiviado, ou devido à inibição dos microrganismos por toxicidade. Quanto ao N-NH4, infere-se que o aumento do TDH proporcionou maior tempo de difusão de oxigênio para o sistema wetland, suprindo a demanda das bactérias heterotróficas, o que possibilitou utilização do oxigênio excedente pelas nitrificantes, culminando na oxidação do N-NH4. / Introduction sewage treatment systems, when applied to the treatment of landfill leachate, have not shown good results. Thus it is necessary to develop alternatives for the treatment of leachate. Objective To assess the post-treatment of landfill leachate by horizontal subsurface flow constructed wetland and moving bed biofilm reactor (MBBR). Materials and methods Two experiments were conducted with treatment of the leachate after removal of ammonia by stripping. In experiment 1, horizontal flow constructed wetlands, at bench scale (total volume of 30.8 L), planted with Cyperus papyrus, Heliconia psittacorum and Gynerium sagittatum, and a control without vegetation, were filled with calcareous gravel. The system was fed with leachate diluted in water with different proportions (between 10 per cent and 30 per cent ) with mean COD concentration of between 336 and 750 mg.L-1 and ammonia nitrogen (NH4-N) between 47 and 199 mg L-1. The operation was divided in three stages: (1st) continuous feeding with mean HRT between 2.7 and 5.3 d; (2nd) regime of effluent recirculation to evaluate the effect of increasing the HRT to 21 days; (3rd) continuous feeding with two wetlands units in series, aimed HRT increasing (between 8.1 and 9.9 d) without recirculation. Experiment 2 was implemented on a pilot scale, with a unit of ammonia stripping followed by MBBR and an anoxic biofilter. The MBBR (380 L working volume) was filled 50 per cent with pall ring. Results and discussion In experiment 1, the COD removal was low, with averages below 40 per cent , with effluent concentration between 270 and 750 mg.L-1. However, there was removal of NH4-N, especially in the 2nd and 3rd stages, with average removal between 43 per cent and 81 per cent , resulting in concentration between 20 and 223 mg.L-1, with HRT influence. In experiment 2, technical problems during the installation and initial operation and interdiction of the campus of USP-Leste disrupted the reactor acclimation and precluded the monitoring for sufficient time to generate consistent data for discussion. Conclusions The low efficiency for COD is probably related to the recalcitrance of the leachate, or due to inhibition of microorganisms by toxicity. For NH4-N, it is inferred that the increase of the HRT provided greater time for diffusion of oxygen to the wetland system, meeting the demand of heterotrophic bacteria, which allowed the use of excess oxygen by nitrifiers, culminating in the oxidation of NH4-N.

Introdução sistemas de tratamento de esgotos, quando aplicados ao tratamento de lixiviado de aterro sanitário, não têm apresentado bons resultados. Por conta disso, faz-se necessário o desenvolvimento de alternativas para o tratamento de lixiviado. Objetivo avaliar o pós-tratamento de lixiviado de aterro sanitário por wetland construído de fluxo subsuperficial horizontal e reator biológico de leito móvel (RBLM). Materiais e métodos foram realizados dois experimentos com tratamento de lixiviado após remoção de amônia por dessorção. No experimento 1 utilizou-se wetlands construídos de fluxo horizontal em escala de bancada (volume total de 30,8 L), plantados com Cyperus papyrus, Heliconia psittacorum e Gynerium sagittatum, e um controle sem vegetação, preenchidos com pedrisco calcário. O sistema foi alimentado com lixiviado diluído em água em diferentes proporções (entre 10 por cento e 30 por cento ) com concentração média de DQO entre 336 e 750 mg.L-1 e nitrogênio amoniacal (N-NH4) entre 47 e 199 mg.L-1. A operação ocorreu em três etapas: (1ª) alimentação contínua e TDH médio entre 2,7 e 5,3 d; (2ª) regime de ciclos de recirculação do efluente, para avaliar o efeito do aumento de TDH para 21 dias; (3ª) alimentação contínua de duas unidades wetlands em série, visando elevar o TDH (entre 8,1 e 9,9 d) sem recirculação. O experimento 2 foi implantado em escala piloto, com uma unidade de dessorção de amônia seguida de um Reator Biológico de Leito Móvel (RBLM) e um biofiltro anóxico. O RBLM (volume efetivo de 380 L) foi preenchido em 50 por cento com anel pall. Resultados e discussão No experimento 1, obteve-se baixa remoção de DQO com médias inferiores a 40 por cento e concentração final entre 270 e 750 mg.L-1. No entanto, verificou-se remoção de N-NH4, principalmente na 2ª e 3ª etapas, com médias de remoção entre 43 por cento e 81 por cento , resultando em concentração de 20 a 223 mg.L-1, com influência do TDH. No experimento 2, problemas técnicos durante a instalação e operação inicial e interdição do campus da USP-Leste prejudicaram a adaptação do reator e impediram o seu monitoramento por tempo suficiente para geração de dados consistentes, sendo interrompido na fase inicial. Conclusões A baixa eficiência para DQO provavelmente está relacionada à recalcitrância do lixiviado, ou devido à inibição dos microrganismos por toxicidade. Quanto ao N-NH4, infere-se que o aumento do TDH proporcionou maior tempo de difusão de oxigênio para o sistema wetland, suprindo a demanda das bactérias heterotróficas, o que possibilitou utilização do oxigênio excedente pelas nitrificantes, culminando na oxidação do N-NH4. / Introduction sewage treatment systems, when applied to the treatment of landfill leachate, have not shown good results. Thus it is necessary to develop alternatives for the treatment of leachate. Objective To assess the post-treatment of landfill leachate by horizontal subsurface flow constructed wetland and moving bed biofilm reactor (MBBR). Materials and methods Two experiments were conducted with treatment of the leachate after removal of ammonia by stripping. In experiment 1, horizontal flow constructed wetlands, at bench scale (total volume of 30.8 L), planted with Cyperus papyrus, Heliconia psittacorum and Gynerium sagittatum, and a control without vegetation, were filled with calcareous gravel. The system was fed with leachate diluted in water with different proportions (between 10 per cent and 30 per cent ) with mean COD concentration of between 336 and 750 mg.L-1 and ammonia nitrogen (NH4-N) between 47 and 199 mg L-1. The operation was divided in three stages: (1st) continuous feeding with mean HRT between 2.7 and 5.3 d; (2nd) regime of effluent recirculation to evaluate the effect of increasing the HRT to 21 days; (3rd) continuous feeding with two wetlands units in series, aimed HRT increasing (between 8.1 and 9.9 d) without recirculation. Experiment 2 was implemented on a pilot scale, with a unit of ammonia stripping followed by MBBR and an anoxic biofilter. The MBBR (380 L working volume) was filled 50 per cent with pall ring. Results and discussion In experiment 1, the COD removal was low, with averages below 40 per cent , with effluent concentration between 270 and 750 mg.L-1. However, there was removal of NH4-N, especially in the 2nd and 3rd stages, with average removal between 43 per cent and 81 per cent , resulting in concentration between 20 and 223 mg.L-1, with HRT influence. In experiment 2, technical problems during the installation and initial operation and interdiction of the campus of USP-Leste disrupted the reactor acclimation and precluded the monitoring for sufficient time to generate consistent data for discussion. Conclusions The low efficiency for COD is probably related to the recalcitrance of the leachate, or due to inhibition of microorganisms by toxicity. For NH4-N, it is inferred that the increase of the HRT provided greater time for diffusion of oxygen to the wetland system, meeting the demand of heterotrophic bacteria, which allowed the use of excess oxygen by nitrifiers, culminating in the oxidation of NH4-N.

This master thesis is based on own studies. A four-month study was performed at Hammarby Sjostad Research Station, which is located in Stockholm. One-stage deammonification process was evaluated in two different system configurations in pilot plant scale. The theoretical background for this thesis works is presented in the first part and where is presented negative impacts of nitrogen compounds in environment and requirements for purified wastewater in European Union (Sweden and Poland). In the next part of the thesis the nitrogen cycle is described and with focus on biological reactions for nitrogen removal. Especially, nitrification/denitrification and anammox processes are described with special focus on parameters affecting the anammox process performance and its advantages and disadvantages of using this process. Experimental results from the four-month study and evaluation of the microbial activity are described in the last part.

Norrvatten produces and delivers drinking water to approximately 700 000 people in the northern part of the Stockholm region in Sweden. In their water treatment plant Görvälnverket, water from Lake Mälaren is purified. During the purification, sludge is produced. To be able to use the sludge as landfilling material, it must be dewatered. Polyacrylamide is used as a flocculant for this purpose. However, in the reject water leaving the sludge and discharged into Lake Mälaren, acrylamide monomers are often left as a rest product from the manufacturing of polyacrylamide. The problem is that acrylamide is toxic to living organisms.  The aim of this master thesis was to evaluate proposed water treatment techniques to reduce acrylamide in the reject water leaving Görvälnverket and reach Norrvatten´s goal of an acrylamide concentration below 0.10 µg/l. The water treatment techniques evaluated were ozonation, moving bed biofilm reactor and trickling filter. Along with that, oxidative stress and genotoxicity from the ozone were analysed. Also, the ability of Lake Mälaren to degrade acrylamide was evaluated. The ozone tests were performed in a pilot plant in a laboratory at IVL Swedish Environmental Research Institute (IVL Svenska Miljöinstitutet), the moving bed biofilm reactor- and the trickling filter test were performed in pilot plants at Görvälnverket and the degradation tests in water from Lake Mälaren were performed at Görvälnverket as well.  The results showed that ozonation of the reject water could reduce acrylamide in the reject water and fulfil Norrvatten´s goal of below 0.10 µg/l acrylamide. When an ozone dose of 0.70 mg/l was applied to the reject water with an acrylamide concentration of 2.5 µg/l, more than 98 % of the acrylamide was reduced. Furthermore, no oxidative stress or genotoxicity seemed to be generated from the ozonation. The moving bed biofilm reactor and the trickling filter did reduce the acrylamide by 80.77 % and 94.7 % respectively and the results suggested that they could be used to reach Norrvatten´s goal. Finally, the results indicated that water from Lake Mälaren could degrade acrylamide at a temperature of 15 °C in 4 days and in 8 °C in 13 days and reach the goal. / Norrvatten producerar och levererar dricksvatten till cirka 700 000 människor i norra delen av Region Stockholm i Sverige. I deras vattenverk Görvälnverket renas vatten från Mälaren. När sjövattnet renas produceras slam. För att slammet ska kunna användas som landfyllnadsmaterial måste det avvattnas. Polyakrylamid används som flockningsmedel i det syftet. I rejektvattnet som lämnar slammet och släpps ut i Mälaren finns ofta akrylamid monomerer kvar som en restprodukt från framställning av polyakrylamid. Problemet med detta är att akrylamid är toxiskt för levande organismer.  Syftet med detta masterexamensarbete var att evaluera föreslagna vattenreningstekniker för att reducera akrylamid i rejektvattnet som lämnar Görvälnverket och inte överstiga en akrylamidkoncentration på 0,10 µg/l som är Norrvattens mål. Vattenreningsteknikerna som evaluerades var ozonering, biofilmreaktor med rörlig bädd och biobädd. Utöver det var oxidativ stress samt genotoxicitet från ozoneringen analyserad. Dessutom var nedbrytningsförmågan av akrylamid i Mälaren testad. Ozonerings försöken utfördes i en pilotanläggning i ett laboratorium hos IVL Svenska Miljöinstitutet, testerna med biofilmreaktor med rörlig bädd och biofilter utfördes i pilotanläggningar på Görvälnverket och nedbrytningstesterna i Mälarvatten utfördes också på Görvälnverket.  Resultaten visade på att ozonering av rejektvattnet kan reducera akrylamid och uppfylla Norrvattens mål om en akrylamidkoncentration på mindre än 0,10  µg/l. När en ozondos på 0,70 mg/l applicerades i rejektvattnet med en akrylamidkoncentration på 2,5 µg/l reducerades mer än 98 % av akrylamiden. Vidare tycktes inte ozoneringen bidra till oxidativ stress eller genotoxicitet. Biofilmreaktorn med rörlig bädd och biofiltret reducerade akrylamid med 80,77 % respektive 94,7 %.  Resultaten visade att dessa tekniker skulle kunna användas för att nå Norrvattens mål. Utöver detta visade studien att vatten från Mälaren kan bryta ner akrylamid i 15 °C på 4 dagar och i 8 °C på 13 dagar och nå målet.

In conventional wastewater treatment plants, methanol, ethanol, and acetate are used as carbon source for the denitrification process in the biological nutrient removal. However, growing concern regarding economical costs and carbon footprints from the fossil-based production of these chemicals have forced the companies to look for other alternatives. VFAs have shown a great potential in replacing the conventionally used carbon sources. If implemented this will result in lower chemical cost and a drastic decrease in carbon footprint as well as striving WWTPs towards sustainable development. In this work denitrification has been analysed using different variations of VFAs such as fermented potato protein liquor, food waste and chicken manure VFA. This was done using a basic laboratory setup of a denitrification reactor which used basic stirring agitation and nitrogen purging to ensure anoxic conditions. Nutrients and excess sCOD were added to ensure the highest denitrification rates. The denitrifying biomass was collected at Gryaab AB in the form of k1-carriers making this process a MBBR. The most influential characteristic of the VFAs is the distribution of the acids in the VFA effluent. Butyric acid along with caproic acid showed the best potential for efficient denitrification. The possibility of concentration of VFA effluent showed a high potential when using a nanofiltration system. A C/N ratio of 4.5 conventionally used when methanol is added showed to be the most optimal condition for VFA addition. The combination of VFAs together with conventional used carbon sources showed the best potential in denitrification efficiency proving to be as good or even better than pure synthetic ones. VFAs effluents showed the best potential in removing the intermediate nitrite from the wastewater at high rates. Overall, VFAs shown a great potential for replacing conventionally used carbon sources, demonstrating the potential of substitution, which if implemented will result in lower carbon footprint and a strive towards sustainable development.

PhD. (Department of Biotechnology, Faculty of Applied and Computer Sciences), Vaal University of Technology. / South Africa utilises more antiretroviral (ARV) compounds per capita than any other nation in the fight against Human Immune Deficiency Virus (HIV) or acquired immunodeficiency syndrome (AIDS). Considering the main entrance pathways of antiviral drugs into the urban water cycle, excretions via urine or faeces from treated individuals play a dominant role. Due to the limited efficiency of conventional biological treatment (activated sludge), ARVs were detected in South African wastewater treatment plant effluents and surface waters. This poses a threat to aquatic environments due to the toxicity of ARVs and can be a potential contributor to ARV resistance due to persistent low level ARV exposure in the general population. This study investigated the efficacy of a moving bed biofilm reactor (MBBR) for ctybtri8nthe elimination of five ARV compounds i.e., tenofovir, emtricitabine, nevirapine, ritonavir and efavirenz from synthetic wastewater. Furthermore, the study also looked at the shift in microbial community compositions of biofilms in the MBBR due to exposure to the ARV compounds. Lastly, the ecotoxicity of the MBBR’s influent and effluent along with the actual ARV compounds were examined. The capacity of ARV degradation by the MBBR was investigated by spiking synthetic wastewater influent with 10 μg/L of five ARV compounds. Actual removal during treatment was assessed by sampling the inlets and outlets of the reactor. A targeted solid phase extraction method with Ultra High Pressure Liquid Chromatography coupled to quadrupole time of flight mass spectrometry (LC-MS/MS) was used to quantify the five ARV compounds. Microbial diversity (alpha-diversity) of seeded sludge from a full-scale municipal WWTP and biofilm samples from a laboratory scale MBBR system during pre- and post-introduction of ARV compounds was investigated by Illumina sequencing of the 16S rRNA gene. Ecological toxicity of the MBBR’s influent and effluent along with the five ARV compounds was determined using the Vibrio fischeri, Daphnia magna and Selenastrum capricornutum toxicity test kits and measured as EC50. After MBBR treatment; Nevirapine, Tenofovir, Efavirenz, Ritonavir and Emtricitabine all showed marked reduction in concentration between the influent and effluent of the MBBR. On average, the percentage removed for Nevirapine, Tenofovir, Efavirenz, Ritonavir and Emtricitabine was 62.31%, 74.18%, 93.62%, 94.18% and 94.87% respectively. Microbial diversity results demonstrated that the introduction of antiretroviral drugs affects the bacterial community composition and diversity considerably. For instance, Nitrosomonas, Nitrospira and Alicycliphilus were found to be higher in post introduction of ARV compounds biofilm samples than in biofilm samples before the introduction of ARV compounds. The EC50 for Tenofovir, Emtricitabine, Nevirapine, Ritonavir and Efavirenz were 82.5, 41.7, 39.3, 60.3 and 0.21 mg/L respectively for S. capricornutum; 81.3, 50.7, 49, 87.1 and 0.43 mg/L respectively for D. magna; and 73.5, 55.1, 41.3, 83.6 and 0.55 mg/L respectively for V. fischeri. The EC50 of the influent and effluent were found to be above 100% concentration, therefore they could not be specifically determined. The ecotoxicity results show that ARV compounds are potentially toxic to the environment, with efavirenz being more toxic than the other four ARV compounds tested. Since there were no toxic effects observed from the effluent, it can be assumed that mineralisation has occurred, or the transformation products are of less or equal toxicity to the influent (because the influent did not show any toxic effects to the model organisms tested).