Resumo Efluentes de um tanque de peixes de cultivo de tilápia do Nilo (Oreochromis niloticus) foram tratados em um sistema de dois reatores anaeróbios de leito fixo paralelos e seguido por um reator agitado suspenso sob ambiente anóxico para remoção de sólidos, matéria orgânica e nitrogênio. A estrutura da comunidade microbiana foi comparada entre os reatores e uma cultura enriquecida de anammox originada de lodo anaeróbio por PCR (Reação em Cadeia de Polimerase) e DGGE (Eletroforese em Gel de Gradiente Desnaturante). A análise de DGGE revelou três grandes aglomerados microbianos, a saber, a biomassa do inóculo, a biomassa nos reatores e a cultura de anammox enriquecida. A similaridade entre a comunidade microbiana recuperada dos reatores anaeróbios e a comunidade anammox foi considerada baixa (32,7%), sugerindo diferenciação da comunidade bacteriana em função da seleção de grupos específicos de anammox. Embora o reator anóxico tenha piorado a qualidade do efluente, os resultados indicam o potencial de remoção de matéria orgânica e amônia em um único reator anaeróbio.

Abstract Effluent from a fish tank with Nile tilapia (Oreochromis niloticus) was treated in a system of two parallel fixed-bed anaerobic reactors followed by a suspended stirred, anoxic reactor for solids, organic matter and nitrogen removal. Microbial community structure was compared between the reactors and an enriched anammox culture originated from anaerobic sludge by using the PCR (Polymerase Chain Reaction) and DGGE (Denaturing Gradient Gel Electrophoresis) techniques. DGGE analysis revealed three large microbial clusters, namely, the biomass in the inoculum, the biomass growing in the reactors, and the enriched anammox culture. The similarity between the microbial community recovered from the anaerobic reactors and the anammox community was considered low (32.7%), suggesting differentiation of the bacterial community as a function of the selection of specific anammox groups. The last step in the anoxic reactor had worsened the effluent quality. The potential of organic matter and ammonia removal in a single anaerobic reactor was demonstrated.

Abstract Mangroves are ecosystems located in the transition zone between land and sea, characterized by periodic flooding that confer to its unique characteristics. Little is known about the transformation of nutrients that occur during the organic matter degradation in this system. In this study, we monitor the nitrogen transformations in soils from three mangroves with distinct levels of contamination using labeled 15NO3-. We also screened the mangroves metagenomes for the presence of genes that encode enzymes involved in denitrification (nirS, nirK, nosZ, norB and narG), anaerobic oxidation of ammonia (anammox) (hh, hao and hzo) and dissimilatory nitrate reduction to ammonium (DNRA) (nrfA). The transformations of 15NO3- indicated the balance of denitrification over anammox and DNRA in all three mangroves, with lower rates of processes in the mangrove affected by oil contamination. The metagenomic analysis detected 56 sequences related to denitrification, 19 with anammox and 6 with DNRA. Genes related with denitrification were phylogenetically distributed among several groups of bacteria (mainly Gammaproteobacteria). Anammox and DNRA related sequences were affiliated with Planctomycetes and Gammaproteobacteria, respectively. Thus, metagenomic and functional approaches supported the description of denitrification, anammox and DNRA rates in mangrove soils, and identified the major bacterial groups involved in these processes.

ABSTRACT The aim of this study was to monitor the stabilization of a sequential batch reactor (SBR) regarding the partial nitritation of poultry slaughterhouse wastewater, aiming at preparing it for a reactor with bacteria that promote anaerobic ammonia oxidation (anammox). A cylindrical reactor with a useful volume of 3.5 L, mechanical agitation (50 rpm), and suspended biomass was used in the experiment. The strategies adopted for nitrite-oxidizing bacteria (NOB) inhibition were cycle time (CT: 24, 16, 6, 5, 4 h and real-time monitoring), free ammonia accumulation, alkalinity restriction and low concentrations of dissolved oxygen (DO). The ammonification process, observed at all experimental phases, negatively influenced the control of partial nitritation process. Alkalinity restriction under low cycle times (4 and 5 h) conditions, together with low dissolved oxygen concentration, was a key factor for nitrite-oxidizing bacteria inhibition. Stability of partial nitritation was only reached by real-time monitoring, being the cycles stopped when 50% N–NH4+ and 50% N–NO2− were present in the reactor.

ABSTRACT This study evaluates the influence of initial nitrogen concentration, temperature and stirring speed on specific anammox activity (SAA). The biomass was tested in single batch reactors with different initial nitrogen concentrations (Assay 1) ranging from 60 to 140 mg Ntotal/L in equimolar ratio (NO2--N/NH4+-N) and in another test to 67.3 mg NH4+-N/L and 92.2 mg NO2--N/L (close to anammox stoichiometric ratio). The anammox biomass was also tested in single batch at different temperatures (from 20 to 37° C) to determine the short-term effects on SAA (Assay 2). In the third assay the stirring speed ranged from 50 to 150 rpm in a sequencing batch reactor (SBR) at 37 ºC. SAA was affected by the stoichiometric molar ratio but not by equimolar initial concentrations. The maximum specific anammox activities were 26.2 mg NH4+-N/g VSS.h in the single batch reactor at 37 ºC with NO2--N/NH4+-N stoichiometric ratio and 33.5 mg NH4+-N/g VSS.h in the SBR at 37 ºC and 50 rpm. The NO2--N/NH4+-N molar ratio affected specific anammox activity, and SAA showed to be more hindered by low increases of stirring speed than reported in the literature.

O aumento da escala de produção agropecuária e agroindustrial traz, como consequência, a necessidade por alternativas para a mitigação dos impactos ambientais. As cadeias produtivas de proteína animal (ex.: suinocultura, bovinocultura e avicultura) produzem efluentes com elevada concentração de compostos nitrogenados. Neste contexto, o processo ANAMMOX (do inglês Anaerobic Ammonium Oxidation) se constitui em uma nova estratégia de alto desempenho visando remover compostos nitrogenados de águas residuárias, agropecuárias e agroindustriais, sob condições quimiolitoautotróficas. Este artigo apresenta uma revisão bibliográfica de trabalhos publicados nos últimos 15 anos sobre estudos deste processo, discutindo sua rota metabólica, demonstrando os micro-organismos envolvidos e os parâmetros de controle do processo, além de estudos desenvolvidos no Brasil e possíveis aplicações.

The increase in agriculture and agro-industrial scale production brings the necessity for alternatives to mitigate environmental impacts. The animal protein production chains (e.g. swine, cattle and poultry) produce effluents with high concentration of nitrogen compounds. In this context, the ANAMMOX (Anaerobic Ammonium Oxidation) process is a new strategy for nitrogen removal in a high rate from agricultural and agro-industrial wastewaters under chemolithoautotrophic conditions. This paper presents a literature review of papers published in the last 15 years about the ANAMMOX process, discussing their metabolic pathway, microorganisms, main control parameters as well as studies conducted in Brazil and application possibilities.

A elevada carga de nitrogênio presente em efluentes da suinocultura é um dos maiores desafios de manejo da atividade. O processo Anammox surge como boa alternativa para a remoção biológica desse nutriente. Este trabalho teve como objetivo aclimatar diferentes amostras de lodos de sistemas de tratamento de efluentes da suinocultura com vistas ao estabelecimento do processo Anammox. Dois inóculos foram coletados na Embrapa Suínos e Aves, Concórdia - SC, um dos quais no fundo de uma lagoa anaeróbia inativa (Inóculo A) e o outro, em um tanque de aeração (Inóculo B). Ambos foram aclimatados até cessar o consumo de N-NO3- e, posteriormente, inoculados em dois reatores (Reator A - Inóculo A e Reator B - Inóculo B). O Reator A apresentou atividade, após aproximadamente 110 dias de operação, enquanto o Reator B precisou de aproximadamente 170 dias, diferença de tempo de partida que pode ser explicada pelas diferentes condições ambientais a que cada lodo estava submetido. As análises de FISH e PCR comprovaram a presença de microrganismos com atividade Anammox, mostrando que lodos de sistemas de tratamento de efluentes da suinocultura podem ser boa fonte de inóculo para o desenvolvimento do processo Anammox.

The high load of nitrogen present in swine wastewater is one of the biggest management challenges of the activity. The Anammox process emerges as a good alternative for biological removal of nitrogen. This study aims to acclimate sludge collected from swine effluent treatment systems to establish the Anammox process. Two sludge samples were collected at Embrapa Swine and Poultry, Concordia - SC, Brazil, one from the bottom of an inactive anaerobic pond (inoculum A) and another from an aeration tank (inoculum B). Both were acclimated until the depletion of NO3-N, being subsequently inoculated in two reactors (Reactor A - Inoculum A and Reactor B - Inoculum B). The Reactor A showed activity after 110 days of operation, while the Reactor B needed 170 days. The difference in the start-up time could be explained by the different environmental conditions to which each sludge was submitted. FISH and PCR analyses confirmed the presence of microorganisms with Anammox activity, demonstrating that the sludge of swine wastewater treatment systems is a good source of inoculum for the development of the Anammox process.

The present study aimed to describe the bacterial community present at an anaerobic up flow bioreactor with ANAMMOX activity, inoculated with the sludge from the anaerobic pond of a swine slurry treatment system. The description was based on the molecular DNA techniques using primers for amplification of complete 16S rRNA gene and also new primers to amplify smaller fragments from 16S rRNA. During the bioreactor operation time, the bacterial community changed significantly, increasing the nitrogen removal efficiency, reaching after 500 days a removal rate of 94%. The complete PCR amplification of 16S rRNA gene generated 17 clones, where three presented similarity with Candidatus Jettenia asiatica (97%), twelve with Janthinobacterium (99%) and two with uncultured clones. The PCR amplification of 436 base pairs had generated 12 clones, of which eight presented 96-100% similarity with Candidatus Anammoxoglobus propionicus, Planctomycete KSU-1 and one with Pseudomonas sp. (99%) and three with uncultured clones.

Bactérias anaeróbias oxidadoras de amônia (bactérias Anammox, do inglês anaerobic ammonium oxidizing bacteria) foram enriquecidas em reator em batelada sequencial (RBS), a partir de lodo proveniente de um sistema convencional de lodos ativados tratando esgoto doméstico de Belo Horizonte (MG). Após três meses de cultivo, atividade Anammox foi detectada no sistema pelo consumo de quantidades estequiométricas de NO2- e NH4+. Análises de hibridação in situ fluorescente (FISH, do inglês fluorescent in situ hybridization) confirmaram a presença de bactérias Anammox, provavelmente Candidatus Brocadia anammoxidans, e revelaram que estas representavam 53% do total de células (após 6 meses de cultivo). O desempenho do reator ao longo dos sete meses de operação demonstrou remoção quase que total de nitrito, baseada em concentração afluente de 61 a 95 mg N-NO2-/L. A eficiência máxima de remoção de amônia alcançada foi de 95%, a partir de concentração afluente de 55 a 82 mg N-NH4+/L.

Anaerobic ammonium-oxidizing (Anammox) bacteria were enriched from sludge collected at a conventional activated sludge system treating domestic wastewater of Belo Horizonte(MG), Brazil, employing a sequencing batch reactor (SBR). After three months of cultivation, Anammox activity was detected in the system by the consumption of stoichiometric amounts of NO2- and NH4+. Fluorescent in situ hybridization (FISH) results revealed the presence of Anammox bacteria (probably Candidatus Brocadia anammoxidans) and showed that they accounted for 53% of the total bacterial population (after 6 months of cultivation). The reactor performance during the seven months of operation showed a near perfect removal of nitrite, based on the influent NO2--N concentration of 61-95 mg/L. The maximum ammonia removal efficiency was 95% from the influent N-NH4+ concentration of 55-82 mg/L.

Ammonium oxidation was thought to be an exclusively aerobic process; however, as recently described in the literature, it is also possible under anaerobic conditions and this process was named ANAMMOX. This work describes the operation of a system consisting of a denitrifying reactor coupled to a nitrifying reactor used for removal of nitrogen from slaughterhouse wastewater. During operation of the denitrifying reactor an average nitrogen ammonium removal rate of 50 mg/Ld was observed. This biomass was used to seed a second reactor, operated in repeated fed batch mode, fed with synthetic medium specific to the growth of bacteria responsible for the ANAMMOX process. The nitrogen loading rate varied between 33 and 67 mgN/Ld and average nitrogen removal was 95% and 40%, respectively. Results of fluorescence in situ hybridization (FISH) confirmed the presence of anammox-like microorganisms in the enriched biomass.