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Fertilizer application and manure use practice in agriculture has become one of the most common sources of dissolved nitrogen species to both ground and surface waters. Nitrogen, released as nitrate (NO3-), ammonium (NH4+) and/or organic nitrogen (DON) is subject to a variety of transformation and attenuation processes in groundwater, including sorption, nitrification, denitrification, dissimilatory nitrate reduction to ammonium (DNRA), ammonification and anaerobic ammonium oxidation (anammox). Of these, only denitrification and anammox represent complete attenuation of nitrogen, releasing nitrogen gas (N2). This study examines the occurrence and mechanisms of nitrogen attenuation in groundwater affected by a manure lagoon. Lagoon effluent is in strong contrast to background water with elevated chemical constituents including NH4+ (mean = 121 mg N/L) and DON (218 mg N/L), which are transported through a fast moving groundwater flow system. The NH4+ rich plume interacts with NO3- rich background water at an interface ~3 m below ground surface. Over 100 m of groundwater transport from the source, total nitrogen (TN) was consistently reduced by 90% over two years of study. This reduction can be largely attributed to dilution (~ 80%), but the remaining 10% reflects a component of nitrogen loss due to attenuation, reflecting 32 mg N/L in attenuation and a TN degradation rate of 0.4 mg/L/day. Localized zones of nitrification and denitrification are evidenced by loss of NO3- accompanied by elevated N2O emissions. Anammox is implicated by localized enrichment of δ15N with according decreases in both NO3- and NH4+ at the plume-background interface and through corroborating microbiological study. Ammonification of DON along the flow path, something not observed in similar studies, is conjectured to have a confounding effect on a detailed isotopic investigation by introducing a second source of NH4+ that is depleted in δ15N-NH4+.

Decades of agricultural use of fertilizer and manure has resulted in nitrogen being the most common groundwater contaminant. Of the known processes for nitrogen attenuation, both denitrification and anammox produce a complete transformation of nitrogen species to dinitrogen gas (N2); however, denitrification is typically also associated with the release of N2O and CO2, both greenhouse gases. Anaerobic ammonium oxidation (Anammox), which has been recently discovered to be more prevalent in groundwater environments than previously thought, simultaneously removes NH4+ and nitrate (NO3-), does not require dissolved organic carbon (DOC), and does not produce greenhouse gas by-products. This study evaluates the natural occurrence of anammox in a manure lagoon plume, as well as the feasibility of enhancing anammox activity by mixing NH4+ rich groundwaters and NO3- rich groundwaters together. Fifteen experiments were undertaken with NH4+-N concentrations ranging between 5-100 mg/L, and a NO3--N ranging from 5-88 mg/L. These experiments suggest a nitrogen removal rate (based on NH4 + removal in anaerobic conditions) from anammox generally in the range of 0.1-0.2 mg/L/day. Based on an absence of dissolved oxygen (DO), and concomitant loss of NO3--N with associated 15N-NO3- enrichment (2.1-8.7‰ ) in 11 experiments, it is considered unlikely that nitrification was the cause of the NH4+ loss observed in these experiments. Concurrent 15NNH4+ enrichment of 4.1-11.5‰ was observed in these 11 experiments. Real-time quantitative polymerase chain reaction (qPCR) DNA analyses were used to show the presence of anammox bacteria and to demonstrate temporal population increases during the experiments (up to 16.3% anammox in total bacteria population) in the three experiments analyzed. Although anammoxrelated N removal rates were modest in these trials, such rates could be significant with respect to the multi-year residence times associated with most groundwater flow systems.

Domestic wastewater is the most abundant type of wastewater. Direct discharge of untreated domestic wastewater has environmental and public health risks due to the presence of organics, nutrients and pathogens. Application of anaerobic processes for the treatment of domestic sewage, which at present is largely treated by aerobic processes, has drawn considerable attention recently. Anaerobic processes can be applied for the removal of organic matter (methanogenesis) and nitrogen (anaerobic ammonium oxidation (Anammox)).The toxicity of fluoride to methanogenesis was investigated. The results indicate that acetoclastic were more susceptible to fluoride than hydrogenotrophic methanogens. The concentration of fluoride causing 50% inhibition (IC50) to acetoclastic ranged from 18.1 to 155.7 mg L-1 while for hydrogenotrophic methanogens was > 400.0 mg L-1.The feasibility of a combined system consisting of anaerobic up-flow anaerobic sludge blanket (UASB) followed by aerobic activated sludge (AS) reactor for removal of carbonaceous and nitrogenous contaminants from strong synthetic sewage (2.5 g chemical oxygen demand (COD) L-1) was also studied. The average combined removal of total COD, volatile fatty acids (VFA) and protein was higher than 89.0%, 99.0% and 97.0%; respectively. Extensive nitrification (96.0%) was observed when dissolved oxygen (DO) concentration was > 2.0 mg L-1. In contrast, only partial nitrification occurred when the AS received high organic loads and/or the DO level was below 2.0 mg L-1.The inhibitory effect of nitrite and nitrate on methanogenesis was evaluated. Methanogenic activity was inhibited by the presence of NOx- compounds (i.e., nitrite and nitrate). The inhibition imparted by nitrate was not due to the nitrate itself, but rather to its reduced intermediate, nitrite. The toxicity of NOx- to methanogens was found to be reversible after all the NOx- were reduced during denitrification.Moreover, the development of Anammox enrichment cultures was evaluated. Anammox cultures were successfully developed using sludge samples collected from municipal wastewater treatment plants (WWTPs) as inocula but not from methanogenic granular sludges. Return activated sludge (RAS) collected from WWTP operating for biological nitrogen removal had the highest intrinsic level of Anammox activity. RAS Anammox culture was developed rapidly within 40 days with a doubling time of 6.8 days.

This research focuses on anoxic ammonium oxidation (anammox). The anammox process for treating high ammonium and low organic carbon wastewater can reduce operational costs to a greater extent than the conventional autotrophic/heterotrophic treatment process can. The process has been widely researched because of its potential economic benefits. However, during long-term reactor operation, sudden reductions of nitrogen removal rates have been reported; maximum nitrogen removal rates in different reactor configurations could not approach values predicted based on mathematical modeling; and the crucial stability parameter, such as nitrite, did not have defined threshold concentration. It was hypothesised that free ammonia (FA) increase is the precursor of the instability of the anammox reactor. If it is true that nitrite up to about 200 mg N/L should stimulate nitrogen removal rate inside of the anammox reactor, when FA is kept below the inhibition threshold concentration. The research presented in the thesis argues that FA plays a larger role than has been previously considered in the instability of the anammox reactor. This study found FA inhibited nitrogen removal rates (NRR) at concentrations exceeding 2 mg N/L. In the pH range 7 to 8, the decrease in anammox activity was independent of pH and related only to the concentration of FA. Nitrite concentrations of up to 200 mg N/L did not negatively affect nitrogen removal rate. This study further found that low nitrite provided stable anammox reactor performance, but that high nitrite was not necessarily the cause for reactor destabilization. During the research high nitrogen removal rate was achieved when low FA was provided. During regular reactor operation at pH 6.5, the NRR at about 6.2 g N/Ld was archived. This value was never achieved before till this study was conducted. Conducted research showed controlling FA at low level is required to approach high rates in anammox reactors. Achieving high rates in anammox reactors allow significant reduction in reactor volume which saves resources. Further studies will be required to identify the FA effect on different microbial interactions, and that may provide more in-depth understanding of the nitrite and FA effect than observations based on NRR alone.

This research focuses on anoxic ammonium oxidation (anammox). The anammox process for treating high ammonium and low organic carbon wastewater can reduce operational costs to a greater extent than the conventional autotrophic/heterotrophic treatment process can. The process has been widely researched because of its potential economic benefits. However, during long-term reactor operation, sudden reductions of nitrogen removal rates have been reported; maximum nitrogen removal rates in different reactor configurations could not approach values predicted based on mathematical modeling; and the crucial stability parameter, such as nitrite, did not have defined threshold concentration. It was hypothesised that free ammonia (FA) increase is the precursor of the instability of the anammox reactor. If it is true that nitrite up to about 200 mg N/L should stimulate nitrogen removal rate inside of the anammox reactor, when FA is kept below the inhibition threshold concentration. The research presented in the thesis argues that FA plays a larger role than has been previously considered in the instability of the anammox reactor. This study found FA inhibited nitrogen removal rates (NRR) at concentrations exceeding 2 mg N/L. In the pH range 7 to 8, the decrease in anammox activity was independent of pH and related only to the concentration of FA. Nitrite concentrations of up to 200 mg N/L did not negatively affect nitrogen removal rate. This study further found that low nitrite provided stable anammox reactor performance, but that high nitrite was not necessarily the cause for reactor destabilization. During the research high nitrogen removal rate was achieved when low FA was provided. During regular reactor operation at pH 6.5, the NRR at about 6.2 g N/Ld was archived. This value was never achieved before till this study was conducted. Conducted research showed controlling FA at low level is required to approach high rates in anammox reactors. Achieving high rates in anammox reactors allow significant reduction in reactor volume which saves resources. Further studies will be required to identify the FA effect on different microbial interactions, and that may provide more in-depth understanding of the nitrite and FA effect than observations based on NRR alone.

Fertilizer application and manure use practice in agriculture has become one of the most common sources of dissolved nitrogen species to both ground and surface waters. Nitrogen, released as nitrate (NO3-), ammonium (NH4+) and/or organic nitrogen (DON) is subject to a variety of transformation and attenuation processes in groundwater, including sorption, nitrification, denitrification, dissimilatory nitrate reduction to ammonium (DNRA), ammonification and anaerobic ammonium oxidation (anammox). Of these, only denitrification and anammox represent complete attenuation of nitrogen, releasing nitrogen gas (N2). This study examines the occurrence and mechanisms of nitrogen attenuation in groundwater affected by a manure lagoon. Lagoon effluent is in strong contrast to background water with elevated chemical constituents including NH4+ (mean = 121 mg N/L) and DON (218 mg N/L), which are transported through a fast moving groundwater flow system. The NH4+ rich plume interacts with NO3- rich background water at an interface ~3 m below ground surface. Over 100 m of groundwater transport from the source, total nitrogen (TN) was consistently reduced by 90% over two years of study. This reduction can be largely attributed to dilution (~ 80%), but the remaining 10% reflects a component of nitrogen loss due to attenuation, reflecting 32 mg N/L in attenuation and a TN degradation rate of 0.4 mg/L/day. Localized zones of nitrification and denitrification are evidenced by loss of NO3- accompanied by elevated N2O emissions. Anammox is implicated by localized enrichment of δ15N with according decreases in both NO3- and NH4+ at the plume-background interface and through corroborating microbiological study. Ammonification of DON along the flow path, something not observed in similar studies, is conjectured to have a confounding effect on a detailed isotopic investigation by introducing a second source of NH4+ that is depleted in δ15N-NH4+.

Decades of agricultural use of fertilizer and manure has resulted in nitrogen being the most common groundwater contaminant. Of the known processes for nitrogen attenuation, both denitrification and anammox produce a complete transformation of nitrogen species to dinitrogen gas (N2); however, denitrification is typically also associated with the release of N2O and CO2, both greenhouse gases. Anaerobic ammonium oxidation (Anammox), which has been recently discovered to be more prevalent in groundwater environments than previously thought, simultaneously removes NH4+ and nitrate (NO3-), does not require dissolved organic carbon (DOC), and does not produce greenhouse gas by-products. This study evaluates the natural occurrence of anammox in a manure lagoon plume, as well as the feasibility of enhancing anammox activity by mixing NH4+ rich groundwaters and NO3- rich groundwaters together. Fifteen experiments were undertaken with NH4+-N concentrations ranging between 5-100 mg/L, and a NO3--N ranging from 5-88 mg/L. These experiments suggest a nitrogen removal rate (based on NH4 + removal in anaerobic conditions) from anammox generally in the range of 0.1-0.2 mg/L/day. Based on an absence of dissolved oxygen (DO), and concomitant loss of NO3--N with associated 15N-NO3- enrichment (2.1-8.7‰ ) in 11 experiments, it is considered unlikely that nitrification was the cause of the NH4+ loss observed in these experiments. Concurrent 15NNH4+ enrichment of 4.1-11.5‰ was observed in these 11 experiments. Real-time quantitative polymerase chain reaction (qPCR) DNA analyses were used to show the presence of anammox bacteria and to demonstrate temporal population increases during the experiments (up to 16.3% anammox in total bacteria population) in the three experiments analyzed. Although anammoxrelated N removal rates were modest in these trials, such rates could be significant with respect to the multi-year residence times associated with most groundwater flow systems.

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Tecnologia, Departamento de Engenharia Civil e Ambiental, Pós-Graduação em Tecnologia Ambiental e Recursos Hídricos, 2015. / Submitted by Fernanda Percia França (fernandafranca@bce.unb.br) on 2016-06-21T17:06:07Z No. of bitstreams: 1 2015_GenildaMariadeOliveira.pdf: 20025076 bytes, checksum: 8405d0acfdc683e08eae68da5444f386 (MD5) / Approved for entry into archive by Raquel Viana(raquelviana@bce.unb.br) on 2016-12-20T14:05:13Z (GMT) No. of bitstreams: 1 2015_GenildaMariadeOliveira.pdf: 20025076 bytes, checksum: 8405d0acfdc683e08eae68da5444f386 (MD5) / Made available in DSpace on 2016-12-20T14:05:13Z (GMT). No. of bitstreams: 1 2015_GenildaMariadeOliveira.pdf: 20025076 bytes, checksum: 8405d0acfdc683e08eae68da5444f386 (MD5) / O controle e tratamento de águas residuárias são os principais mecanismos para evitar o aporte de nitrogênio aos ecossistemas aquáticos. Esse nutriente em altas concentrações ocasiona problemas ambientais e de saúde pública. O processo anammox, realizado por bactérias anaeróbias que oxidam a amônia usando o nitrito como aceptor final de elétrons, apresenta-se com potencial promissor para a remoção de nitrogênio de águas residuárias, devido à redução dos custos comparado ao processo convencional de tratamento, composto pelas etapas de nitrificação e desnitrificação heterotrófica. No Brasil, deve-se buscar fontes de inóculos e estudar as condições para o estabelecimento do processo anammox para quando esse mecanismo de remoção de nitrogênio tornar-se uma alternativa de tratamento viável tecnicamente. Nesse sentido, foram objetivos desse trabalho fornecer informações sobre fontes de inóculos, enriquecimento de micro-organismos e ativação de biomassa para partida de sistemas em bateladas visando remoção biológica de amônia pelo processo anammox. Foram testadas 6 fontes de inóculos, oriundas de lodos de sistemas tratando efluentes de abatedouros de bovinos e suínos, sedimentos do Lago Paranoá e da lagoa de acumulação de chorume do aterro controlado Jóckey Clube de Brasília DF, em reatores operados por bateladas em escala de bancada, usando água residuária sintética acrescida de concentrações crescentes dos substratos amônia e nitrito. Durante 120 dias, os sistemas foram ajustados para promoção do enriquecimento da biomassa anammox. Posteriormente testaram-se os sistemas para avaliação da atividade anammox. Sugere-se a presença de micro-organismos anammox, verificados por PCR e por sequenciamento dos fragmentos amplificados dos genes para o RNAr 16S, nas biomassas provenientes do sedimento do lago Paranoá, Brasília DF, dos lodos das lagoas anaeróbia e facultativas tratando efluentes de abatedouros. Ocorreu o aumento da concentração de cópias de fragmentos do gene para RNAr 16S, avaliados por PCRq após os 120 dias, indicando o enriquecimento da biomassa anammox. As condições estabelecidas para os sistemas biológicos parecem selecionar o mesmo grupo de bactérias anammox, independentemente da origem da biomassa, possivelmente havendo substituição de dominância dos grupos de bactérias anammox inicialmente presentes. Os índices de similaridades, calculados por comparações entre as sequências de nucleotídeos obtidas nesse estudo com aquelas de candidatas espécies anammox depositadas no Genbank utilizando o BLAST, indicam que os micro-organismos presentes nos sistemas testados pertencem à Ordem Brocadiales. Limitação de nitrito pode interferir no sucesso do processo, uma vez que as bactérias anammox têm menor afinidade por esse substrato do que as nitratantes e heterotróficas desnitrificantes. Em todos os sistemas biológicos testados o metabolismo anammox não foi exclusivo, uma vez que não foram atingidos os índices estequiométricos esperados. Nos sistemas biológicos ocorreu a concomitância dos processos metabólicos anammox, nitritação, nitratação e desnitrificação heterotrófica. Os inóculos utilizados e/ou as condições estabelecidas nesse estudo não favoreceram a redução do tempo de partida nos sistemas biológicos, sendo necessários mais de 200 dias de operação para enriquecimento e estabilidade do processo anammox. / The control and treatment of wastewater are the main mechanisms to prevent the nitrogen supply to aquatic ecosystems. This nutrient in high concentrations causes environmental and public health problems. The anammox process, carried out by anaerobic bacteria that oxidize ammonia using nitrite as a final electron acceptor, is presented with promising potential for the removal of nitrogen from wastewater, due to its reduced costs compared to the conventional treatment process, which is composed of nitrification and heterotrophic denitrification stages. In Brazil, one must seek sources of inoculum and study the conditions for the establishment of anammox process when this nitrogen removal mechanism becomes a technically viable treatment alternative. In this sense, the objectives of this study were to provide information on sources of inoculum, enrichment of microorganisms and activation of biomass for the startup of batch systems targeting biological removal of ammonia by anammox process. Six sources of inoculum were tested, derived from sludge systems treating wastewater from cattle and pig slaughterhouses, sediments from Paranoa Lake and from the leachate accumulation lake of the controlled landfill Jockey Club of Brasilia DF, in reactors operated by batch in bench scale, using synthetic wastewater together with increasing concentrations of substrates ammonia and nitrite. For 120 days, the systems were adjusted for promoting the enrichment of anammox biomass. Afterwards, the systems for evaluation of anammox activity were tested. It is suggested the presence of anammox microorganisms, checked by PCR and sequencing of the amplified fragments of the genes for 16S rRNA from biomass in the lake Paranoa sediment, Brasilia DF, the sludge of anaerobic and facultative lagoons treating effluent from slaughterhouse. There was an increase in the concentration of copies of gene fragments for 16S rRNA, evaluated by PCRq after 120 days, indicating enrichment of anammox biomass. The conditions developed for the biological systems seem to select the same group of anammox bacteria, regardless of their biomass origin, possibly having replacement of the anammox bacteria groups initially present. The levels of similarity, calculated by comparing the nucleotide sequences obtained in this study with those of candidate species anammox deposited in Genbank using the BLAST, indicate that microorganisms present in the tested systems belong to the Order Brocadiales. Limitation of nitrite may interfere with the success of the process, since the anammox bacteria has lower affinity for this substrate than nitrite oxidiziers and heterotrophic denitrifiers. In all the biological systems that were tested, anammox metabolism was not unique, since the expected stoichiometric ratio was not achieved. In the biological systems, there was the concurrence of anammox metabolic processes, nitritation, nitration and heterotrophic denitrification. The inoculum used and / or the conditions established in this study did not favor the reduction of the startup time of biological systems, requiring more than 200 days of operation for enrichment and stability of the anammox process.

Esta pesquisa objetivou enriquecer biomassa capaz de realizar a oxidação anaeróbia do amônio (anammox) utilizando inóculo proveniente de reator nitrificante-desnitrificante, com a finalidade de estabelecer biofilme nitrificante-anammox em reator de leito fixo. O enriquecimento foi realizado em reator operado em bateladas sequenciais (RBS), com volume útil de 5 L e tempo de ciclo, inicialmente, de 56 h, e depois, sem tempo predeterminado (estratégias I e II). Após 89 dias de operação, 27,2 mg de \'N\'-\'N0 IND.2\'POT.-\'/L e 32,1 mg de \'N\'-\'NH IND.4\'POT.+\'/L foram consumidos concomitantemente. A estratégia III consistiu de batelada alimentada com ciclos de sete dias com afluente contendo 210 mg de cada composto nitrogenado. Na última estratégia (IV) a operação foi com ciclos de 24 h. Nessa etapa, a carga nitrogenada aplicada (CNA) foi aumentada de 155 g de \'N\' (\'N\'-\'N0 IND.2\'POT.-\' \'N\'-\'NH IND.4\'POT.+\')/\'M POT.3\' dia para 1.405,7 g de \'N\'/\'M POT.3\' dia com eficiências de conversão de nitrogênio de 91,7% e 98,0%, respectivamente. Essa biomassa foi inoculada em reator de leito fixo ascendente (RLF) visando estabelecimento da biomassa anammox em meio suporte (PEBD). Sob tais condições foi obtido eficiência de conversão de nitrogênio de 97,6% e carga nitrogenada removida média de 598,5 \'+ OU -\' 22,5 g \'N\'/\'M POT.3\' dia. Após estabelecimento de biomassa anammox, foi adicionado lodo ativado da indústria Volkswagen (São Carlos-SP) para formação de biofilme nitrificante-anammox. Nessa fase, a remoção de nitrogênio foi de 19,2% para CNA de 112,2 g \'N\'/\'M POT.3\' dia. A atividade anammox específica máxima foi 33,5 mg \'N\'-\'NH IND.4\'POT.+\'/g SSV h com a biomassa submetida à 50 rpm. Paralelamente ao processo de enriquecimento, foi verificada a influência de micronutrientes em condições nitrificantes em três quimiostatos, nas seguintes condições: Q1 alimentado com meio contendo solução de micronutrientes completa, Q2 alimentado sem solução de micronutrientes e Q3 alimentado com solução de micronutrientes sem o elemento Boro (quimiostato experimental). Nas três condições a estabilidade foi atingida com 11 dias de operação com conversão média de nitrogênio amoniacal de 99 \'+ OU -\' 1,5%, 94,6\'+ OU -\' 6,3% e 93,3\'+ OU -\' 7,3%, para Q1, Q2 e Q3, respectivamente, para 79 mg \'N\'-\'NH IND.4\'POT.+\'/L afluente. Após 450 dias de operação do RBS foi constatado semelhança do clones com Brocadia anammoxidans, Planctomycetes, Proteobacteria, Chlorobi, Nitrospira, filo Chloroflexi e ao filo candidato OP 11. A composição microbiana encontrada no RLF com 139 dias de operação (final da fase anammox) foi de 48% dos clones relacionados à B. anammoxidans, 4% relacionados à Planctomycetes não cultivados, 12% relacionados à Proteobacteria, 8% relacionados à Chlorobi, 24% relacionados à Nitrospira, 4% relacionados ao filo Chloroflexi. Pode-se concluir que a biomassa aderida em PEBD selecionou positivamente microrganismos anammox e Nitrospira e negativamente aos filamentos relacionados ao filo Chloroflexi. / This research aimed to enhance biomass capable of performing anaerobic ammonia oxidation (anammox) using inoculum from nitrifying-denitrifying reactor, with the goal of establishing nitrifying-anammox biofilm in fixed bed reactor. The enrichment was performed in sequencing batch reactor (SBR), with a volume of 5 L and cycle time, initially, 56 h, and then, without pre-set time (strategies I and II). After 89 operation days, 27.2 mg \'N\'-\'N0 IND.2\'POT.-\'/L and 32.1 mg \'N\'-\'NH IND.4\'POT.+\'/L were consumed concomitantly. The strategy consisted of fed batch III with seven days cycles with influent containing 210 mg of each nitrogen compound. The last strategy (IV) was with 24 h/cycle. At this strategy, the nitrogen applied load (NAL) was increased from 155 \'N\' (\'N\'-\'N0 IND.2\'POT.-\' + \'N\'-\'NH IND.4\'POT.+\')/\'M POT.3\' to 1405.7 g \'N\'/\'M POT.3\' day with conversion efficiencies of 91.7% nitrogen and 98.0%, respectively. This biomass was inoculated into fixed bed reactor up (FBR) in order to establish the anammox biomass in support medium (LDPE). Under such conditions was obtained nitrogen conversion efficiency of 97.6% and nitrogen load removed an average of 598.5 \'+ OU -\' 22.5 g \'N\'/\'M POT.3\' day. After establishment of anammox biomass it was added activated sludge - Volkswagen industry (São Carlos-SP) - for nitrifying-anammox biofilm. At that stage the removaI of nitrogen was 19.2% to 112.2 g CNA \'N\'/\'M POT.3\' day. Simultaneously to enrichment process, was verified the influence of micronutrients in nitrifying conditions in three chemostats, as follows: Q1 fed with medium containing micronutrients solution complete feeds without Q2 and Q3 micronutrients solution fed micronutrients solution without the element Boron (chemostat experiment). In the three conditions stability was achieved with 11 days of operation with average conversion of ammonia nitrogen of 99 \'+ OU -\' 1.5%, 94.6 \'+ OU -\' 6.3% and 93.3 \'+ OU -\' 7.3% for Q1, Q2 and Q3 respectively for 79 mg \'N\'-\'NH IND.4\'POT.+\'/L. After 450 days of operation of the RBS was found similarity of clones with Brocadia anammoxidans, Planctomycetes, Proteobacteria, Chlorobi, Nitrospira, Chloroflexi phyla and candidate phylum OP 11. The microbial composition found in the FBR with 139 days of operation (end of anammox phase) was 48% of clones related to B. anammoxidans, 4% related to uncultured Planctomycetes, Proteobacteria related to 12%, 8% related to Chlorobi, 24% related to Nitrospira, 4% related to the phylum Chloroflexi. It can be concluded that biomass adhered to LDPE selected anammox microorganisms and Nitrospira positively, and negatively to the filaments related to the Chloroflexi phylum.