Enhanced nitrogen removal by bioelectrochemical coupling anammox and characteristics of microbial communities / 生物工程学报

To investigate the bioelectrochemical enhanced anaerobic ammonia oxidation (anammox) nitrogen removal process, a bioelectrochemical system with coupled anammox cathode was constructed using a dual-chamber microbial electrolysis cell (MEC). Specifically, a dark incubation batch experiment was conducted at 30 ℃ with different influent total nitrogen concentrations under an applied voltage of 0.2 V, and the enhanced denitrification mechanism was investigated by combining various characterization methods such as cyclic voltammetry, electrochemical impedance spectroscopy and high-throughput sequencing methods. The results showed that the total nitrogen removal rates of 96.9%±0.3%, 97.3%±0.4% and 99.0%±0.3% were obtained when the initial total nitrogen concentration was 200, 300 and 400 mg/L, respectively. In addition, the cathode electrode biofilm showed good electrochemical activity. High-throughput sequencing results showed that the applied voltage enriched other denitrifying functional groups, including Denitratisoma, Limnobacter, and ammonia oxidizing bacteria SM1A02 and Anaerolineaceae, Nitrosomonas europaea and Nitrospira, besides the anammox bacteria. These electrochemically active microorganisms comprised of ammonium oxidizing exoelectrogens (AOE) and denitrifying electrotrophs (DNE). Together with anammox bacteria Candidatus Brocadia, they constituted the microbial community structure of denitrification system. Enhanced direct interspecies electron transfer between AOE and DNE was the fundamental reason for the further improvement of the total nitrogen removal rate of the system.

Denitrifying phosphate accumulating organisms and its mechanism of nitrogen and phosphorus removal / 生物工程学报

Water eutrophication poses great threats to protection of water environment. Microbial remediation of water eutrophication has shown high efficiency, low consumption and no secondary pollution, thus becoming an important approach for ecological remediation. In recent years, researches on denitrifying phosphate accumulating organisms and their application in wastewater treatment processes have received increasing attention. Different from the traditional nitrogen and phosphorus removal process conducted by denitrifying bacteria and phosphate accumulating organisms, the denitrifying phosphate accumulating organisms can simultaneously remove nitrogen and phosphorus under alternated anaerobic and anoxic/aerobic conditions. It is worth noting that microorganisms capable of simultaneously removing nitrogen and phosphorus absolutely under aerobic conditions have been reported in recent years, but the mechanisms remain unclear. This review summarizes the species and characteristics of denitrifying phosphate accumulating organisms and the microorganisms capable of performing simultaneous nitrification-denitrification and phosphorous removal. Moreover, this review analyzes the relationship between nitrogen removal and phosphorus removal and the underlying mechanisms, discusses the challenges of denitrifying phosphorus removal, and prospects future research directions, with the aim to facilitate process improvement of denitrifying phosphate accumulating organisms.

Mechanism of trehalose-enhanced metabolism of heterotrophic nitrification-aerobic denitrification community under high-salt stress / 生物工程学报

Heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria are aerobic microorganisms that can remove nitrogen under high-salt conditions, but their performance in practical applications are not satisfactory. As a compatible solute, trehalose helps microorganisms to cope with high salt stress by participating in the regulation of cellular osmotic pressure, and plays an important role in promoting the nitrogen removal efficiency of microbial populations in the high-salt environment. We investigated the mechanism of exogenous-trehalose-enhanced metabolism of HN-AD community under high-salt stress by starting up a membrane aerobic biofilm reactor (MABR) to enrich HN-AD bacteria, and designed a C150 experimental group with 150 μmol/L trehalose addition and a C0 control group without trehalose. The reactor performance and the community structure showed that NH4+-N, total nitrogen (TN) and chemical oxygen demand (COD) removal efficiency were increased by 29.7%, 28.0% and 29.1%, respectively. The total relative abundance of salt-tolerant HN-AD bacteria (with Acinetobacter and Pseudofulvimonas as the dominant genus) in the C150 group reached 66.8%, an 18.2% increase compared with that of the C0 group. This demonstrated that trehalose addition promoted the enrichment of salt-tolerant HN-AD bacteria in the high-salt environment to enhance the nitrogen removal performance of the system. In-depth metabolomics analysis showed that the exogenous trehalose was utilized by microorganisms to improve proline synthesis to increase resistance to high-salt stress. By regulating the activity of cell proliferation signaling pathways (cGMP-PKG, PI3K-Akt), phospholipid metabolism pathway and aminoacyl-tRNA synthesis pathway, the abundances of phosphoethanolamine, which was one of the glycerophospholipid metabolites, and purine and pyrimidine were up-regulated to stimulate bacterial aggregation and cell proliferation to promote the growth of HN-AD bacteria in the high-salt environment. Meanwhile, the addition of trehalose accelerated the tricarboxylic acid (TCA) cycle, which might provide more electron donors and energy to the carbon and nitrogen metabolisms of HN-AD bacteria and promote the nitrogen removal performance of the system. These results may facilitate using HN-AD bacteria in the treatment of high-salt and high-nitrogen wastewater.

Research progress of combined anaerobic ammonium oxidation (ANAMMOX) process for nitrogen removal / 生物工程学报

Anaerobic ammonia oxidation (ANAMMOX) process is an efficient and low-cost biological nitrogen removal process. However, it still faces some challenges in mainstream applications due to the limitation of substrate types and nitrate accumulation. In recent years, the combined process of anammox has been widely studied to solve the above problems. In this paper, the combined processes of anammox developed in recent years are reviewed, and discussed from the process principle, advantages and disadvantages, influencing factors, process extensibility and the key bottlenecks existing in the promotion and application, as well as the relevant work of the subject group. Finally, we take an outlook on the development of the combined anaerobic ammonia oxidation process in municipal domestic wastewater treatment.

Craft Brewery Wastewater Treatment: a Fixed-Bed Single-Batch Reactor with Intermittent Aeration to Remove COD and TN

Abstract This study evaluated an intermittently aerated, fixed-bed, single-batch reactor, with mini BioBob© as biofilm media support, as an alternative treatment of craft brewery wastewater. In order to remove chemical oxygen demand (COD) and total nitrogen (TN), seven conditions were performed in a central composite experimental design (CCD) with different aeration times (1, 2 and 3 h in a 4 h cycle) and hydraulic retention times (HRT) (12, 16 and 20 h). The results showed that the removal of COD and TN were positively affected by increased aeration time and HRT. The condition that presented the best quality effluent was Condition No. 1 (20 h HRT and 3 h aeration), with 209 ± 28 mg COD L-1; 3.00 ± 0.15 mg TKN L-1 ; and 0.67 ± 0.11 mg NO3-N L-1. Kinetic assays showed that the highest values for the substrate removal rate constant, kCOD = 0.1774 h-1 were obtained with the longest aeration time (3 h). The most probable number (MPN) test showed a higher concentration of denitrifying bacteria (heterotrophic), 3.3 x 106, than for AOB and NOB bacteria (autotrophic), which were 4.9 x 103 and 2.7 x 103, respectively. Moreover, it was possible to verify that correcting the influent alkalinity with 7.14 mg CaCO3 for each 1 mg of TKN resulted in better process efficiency. It was concluded that COD and TN can be removed from craft brewery wastewater using an intermittently aerated, fixed-bed, single-batch reactor with mini Biobob© as biofilm media support.

Effects of Fe₃O₄ on the denitrification performance of Pseudomonas stutzeri / 生物工程学报

Biological denitrification is the most widely used technology for nitrate removal in wastewater treatment. Conventional denitrification requires long hydraulic retention time, and the nitrate removal efficiency in winter is low due to the low temperature. Therefore, it is expected to develop new approaches to enhance the denitrification process. In this paper, the effect of adding different concentrations of Fe₃O₄ nanoparticles on the denitrification catalyzed by Pseudomonas stutzeri was investigated. The maximum specific degradation rate of nitrate nitrogen improved from 18.0 h⁻¹ to 23.7 h⁻¹ when the concentration of Fe₃O₄ increased from 0 mg/L to 4 000 mg/L. Total proteins and intracellular iron content also increased along with increasing the concentration of Fe₃O₄. RT-qPCR and label-free proteomics analyses showed that the relative expression level of denitrifying genes napA, narJ, nirB, norR, nosZ of P. stutzeri increased by 55.7%, 24.9%, 24.5%, 36.5%, 120% upon addition of Fe₃O₄, and that of denitrifying reductase Nap, Nar, Nir, Nor, Nos increased by 85.0%, 147%, 16.5%, 47.1%, 95.9%, respectively. No significant difference was observed on the relative expression level of denitrifying genes and denitrifying reductases between the bacteria suspended and the bacteria adhered to Fe₃O₄. Interestingly, the relative expression level of electron transfer proteins of bacteria adhered to Fe₃O₄ was higher than that of the bacteria suspended. The results indicated that Fe₃O₄ promoted cell growth and metabolism through direct contact with bacteria, thereby improving the denitrification. These findings may provide theoretical support for the development of enhanced denitrification.

Advances in denitrification microorganisms and processes / 生物工程学报

Denitrification is an indispensable part of most sewage treatment systems. The biological denitrification process has attracted much attention in the past decades due to the advantages such as cost-effectiveness, process simplicity, and absence of secondary pollution. This review summarized the advances on biological denitrification processes in recent years according to the different physiological characteristics and denitrification mechanisms of denitrification microorganisms. The pros and cons of different biological denitrification processes developed based on nitrifying bacteria, denitrifying bacteria, and anaerobic ammonia-oxidizing bacteria were compared with the aim to identify the best strategy for denitrification in a complex wastewater environment. The rapid development of synthetic biology provides possibilities to develop highly-efficient denitrifying strains based on mechanistic understandings. Combined with the applications of automatic simulation to obtain the optimal denitrification conditions, cost-effective and highly-efficient denitrification processed can be envisioned in the foreseeable future.

Diversity and abundance of denitrifiers during cow manure composting / Diversidad y abundancia de desnitrificadores durante el compostaje del estiércol de vaca

Diversity and abundance of the denitrifying genes nirK, nirS and nosZ were investigated in cow manure compost using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and real-time quantitative PCR (qPCR), respectively. These three genes were detected in all the stages of the composting process. Phylogenetic analysis showed that the nirK gene was closely related to Rhizobiales, Burkholderiales, the nirS gene was closely related to Pseudomonadales and Burkholderiales, and the nosZ gene was closely related to Rhodospirillales, Rhizobiales, Pseudomonadales, and Alteromonadales. qPCR results showed that the abundance of these three genes (nirK, nirS and nosZ) reached the peak value in the late thermophilic stage of composting and abundance of the nirK gene was higher than that of the nosZ gene and the nirS gene. Redundancy analysis (RDA) showed that the diversity of the nirK and nirS genes was significantly correlated with ammonium (p < 0.05), the diversity of the nosZ gene was significantly correlated with pH (p < 0.05) and the abundance of the nirK nirS and nosZ genes was significantly correlated with temperature (p< 0.05).

La diversidad y la abundancia de los genes desnitrificadores nirK, nirS, nosZ en el compost de estiércol de vaca se investigaron por medio de la reacción en cadena de la polimerasa seguida de electroforesis en gel con gradiente de desnaturalización (PCR-DGGE) y por PCR cuantitativa (qPCR) en tiempo real, respectivamente. Estos 3 genes fueron detectados durante todas las fases del compostaje. El análisis filogenético mostró estrecha relación del gen nirK con Rhizobiales y Burkholderiales, del gen nirS con Pseudomonadales y Burkholderiales y del gen nosZ con Rhodospirillales, Rhizobiales, Pseudomonadales y Alteromonadales. Los resultados de la qPCR mostraron que la abundancia de los genes nirK, nirSy nosZ alcanzó el valor máximo en la fase termofílica tardía del compostaje, y que la abundancia del gen nirK era más elevada que los de los genes nosZ y nirS. El análisis de redundancia (RDA) mostró que la diversidad de los genes nirK y nirS estaba significativamente correlacionada con la concentración de amonio (p<0,05), mientras que la del gen nosZ estaba significativamente correlacionada con el pH (p<0,05). También mostró que la abundancia de los genes nirK, nirS y nosZ estaba significativamente correlacionada con la temperatura (p<0,05).

Advances in heterotrophic nitrification-aerobic denitrifying bacteria for nitrogen removal under extreme conditions / 生物工程学报

Heterotrophic nitrification-aerobic denitrification (HN-AD) is an enrichment and breakthrough theory of traditional autotrophic nitrification heterotrophic denitrification. Heterotrophic nitrification-aerobic denitrifiers with the feature of wide distribution, strong adaptability and unique metabolic mechanism have many special advantages, including fast-growing, rapid biodegradability and long lasting activity, which can rapidly remove ammonia nitrogen, nitrate nitrogen (NO₃⁻-N) and nitrite nitrogen (NO₂⁻-N) under aerobic conditions simultaneously. Therefore, HN-AD bacteria show the important potential for denitrification under extreme conditions with high-salt, low-temperature or high-ammonia nitrogen environment, and HN-AD bacteria attract extensive attention in the field of biological denitrification of wastewater. In this review, we first introduce the previously reported HN-AD bacterial species which have denitrification performance in the extreme environments and state their typical metabolic mechanism. Then, we systematically analyze the nitrogen removal characteristics and potential under extreme conditions. We also briefly describe the progress in the application of HN-AD bacterial. Finally, we outlook the application prospects and research directions of HN-AD denitrification technology.

Emisión de óxido nitroso, nitrificación, desnitrificación y mineralización de nitrógeno durante el cultivo del arroz en 2 suelos de Uruguay / Nitrous oxide emission, nitrification, denitrification and nitrogen mineralization during rice growing season in 2 soils from Uruguay

Los procesos microbianos como la mineralización, la nitrificación y la desnitrificación regulan la dinámica del nitrógeno en el suelo. Estos 2 últimos son los principales responsables de la emisión de óxido nitroso (N2O). En este trabajo se determinaron los flujos de N2O en momentos clave del ciclo de cultivo del arroz en 2 sitios que diferían principalmente en el contenido de materia orgánica del suelo (MO), en las localidades de Salto (mayor MO) y de Treinta y Tres. Dichos momentos clave fueron a la siembra, en macollaje, en primordio floral y a la madurez. También se determinó el potencial de mineralización neta de N y las actividades y los números más probables (NMP) de oxidantes de NH4+ y de desnitrificantes. El potencial de mineralización de N, así como la actividad y el NMP de oxidantes de NH4+, no variaron con el tipo de suelo. Sin embargo, la actividad y el NMP de desnitrificantes fueron mayores en el suelo con mayor contenido de MO, independiente de la etapa del cultivo. A su vez, en las etapas finales del ciclo del cultivo, el NMP de desnitrificantes aumentó coincidiendo con el mayor potencial de mineralización y el mayor contenido de N mineral del suelo. Solo se observó un incremento en el flujo de N2O en el suelo de Salto a la madurez del arroz y cuando el suelo ya había sido drenado (44,2g N-N2O/ha d, frente a 20,8g N-N2O/ha d en Treinta y Tres). Esta investigación señala la importancia de estudiar las emisiones en distintos tipos de suelos y de continuar la medición luego del drenaje del cultivo de arroz para la elaboración de los inventarios de gases de efecto invernadero.

Microbial processes such as mineralization, nitrification and denitrification regulate nitrogen dynamics in the soil. The last two processes may produce nitrous oxide (N2O). In this work N2O fluxes were quantified at four moments of the rice cycle, sowing, tillering, panicle initiation and maturity, in two sites that differed mainly in their soil organic matter (OM) content, Salto (higher OM) and Treinta y Tres. Potential net N mineralization, ammonium oxidation and denitrification as well as the most probable numbers (MPN) of ammonia oxidizers and denitrifiers were determined. Potential N mineralization did not vary with the soil type and increased at rice maturity. Neither ammonia oxidation potential nor MPN were different among the soils. However, the soil with higher OM exhibited higher activity and MPN of denitrifiers, irrespective of the rice stage. In turn, at the latest phases of the crop, the MPN of denitrifiers increased coinciding with the highest mineralization potential and mineral N content of the soil. Significant differences in N2O flux were observed in Salto, where the highest emissions were detected at rice maturity, after the soil was drained (44.2 vs 20.8g N-N2O/ha d in Treinta y Tres). This work shows the importance of considering the soil type and end-of-season drainage of the rice field to elaborate GHGs (greenhouse gases) inventories.

Novos reguladores de resposta envolvidos na virulência de Pseudomonas aeruginosa / New response regulators involved in Pseudomonas aeruginosa virulence

Os sistemas de sinalização de dois componentes são sistemas prevalentes em bactérias, permitindo a adaptação a diferentes condições ambientais. O sistema de dois componentes classicamente possui uma proteína histidina quinase, o primeiro componente, capaz de reconhecer o estímulo ambiental e fosforilar o regulador de resposta, o segundo componente. Pseudomonas aeruginosa é uma proteobactéria ubíqua, capaz de infectar hospedeiros filogeneticamente distintos. Esse patógeno oportunista apresenta um dos maiores conjuntos de sistemas de dois componentes em bactérias, que permite que ela sobreviva numa grande gama de ambientes, incluindo humanos. P. aeruginosa UCBPP-PA14 apresenta pelo menos 64 histidina quinases e 76 reguladores de resposta codificados em seu genoma. Diversos sistemas de dois componentes já foram correlacionados com a virulência, sendo o sistema GacSA o exemplo melhor caracterizado. Há poucos estudos sistemáticos sobre o envolvimendo dos reguladores de resposta na virulência de P. aeruginosa e os sinais que induzem a ativação dos reguladores de resposta precisam ser encontrados. Para identificar novos reguladores de resposta envolvidos na patogenicidade, infecções in vitro em macrófagos e in vivo em Drosophila melanogaster foram realizadas neste trabalho. Os macrófagos foram infectados com cada mutante dos reguladores de resposta ou com a linhagem selvagem, e a produção da citocina pró-inflamatória TNF-α e o clearance bacteriano foram determinados. Alternativamente, as moscas foram infectadas utilizando-se a estratégia de feeding e a sobrevivência foi verificada. Utilizando-se essas abordagens, a identificação de diversos reguladores de resposta com papel na virulência foi alcançada, além de se corfirmar o papel de reguladores de resposta já estudados. Um dos novos genes envolvidos em virulência, PA14_26570 (nomeado neste trabalho de atvR), codifica um regulador de resposta atípico com substituição no aspartato fosforilável para glutamato, o que usualmente induz um estado sempre ativo. Um mutante não polar em atvR foi construído e macrófagos infectados com a linhagem ΔatvR confirmaram um maior clearance bacteriano e maior produção de TNF-α em comparação aos macrófagos infectados com a linhagem selvagem. Para comprovar a participação de AtvR durante a patogênese, um modelo de pneumonia aguda em camundongos foi utilizado. Camundongos infectados com a linhagem ΔatvR apresentaram uma maior sobrevivência em comparação aos camundongos infectados com a linhagem selvagem. Além disso, os camundongos infectados com ΔatvR apresentaram menor carga bacteriana, aumento no recrutamento de neutrófilos ativados e aumento na produção de citocinas pró-inflamatórias (TNF-α e IFN-γ). Utilizando-se uma abordagem transcritômica (RNA-Seq), foi determindo diversos genes são regulados positivamente na linhagem superexpressando AtvR em relação à linhagem controle. Dentre esses, os clusters de respiração anaeróbia nar, nir, nor e nos estão incluídos. Esse resultado foi confirmado por qRT-PCR e análises fenotípicas, em que a linhagem ΔatvR apresentou menor crescimento e expressão da nitrato redutase durante condições de hipóxia em comparação à linhagem selvagem. Em suma, neste trabalho foi demonstrado que diversos reguladores de resposta são importantes para a virulência de P. aeruginosa em macrófagos in vitro e in vivo em Drosophila, além de caracterizar o regulador de resposta atípico AtvR, que regula a respiração anaeróbica por desnitrificação, permitindo que P. aeruginosa possa infectar e colonizar o hospedeiro com maior eficiência

Two-component systems are widespread in bacteria, allowing the adaptation to environmental changes. A two-component system is classically composed by a sensor kinase that phosphorylates a cognate response regulator. Pseudomonas aeruginosa is a ubiquitous proteobacterium able to cause disease in several hosts. This opportunistic pathogen presents one of the largest sets of two-component systems known in bacteria, which certainly contributes to its ability to thrive in a wide range of environmental settings, including humans. P. aeruginosa UCBPP-PA14 genome codes for at least 64 sensor kinases and 76 response regulators. Some response regulators are already known to be related to virulence, with the GacSA system as the best characterized. There are no systematic studies about the involvement of P. aeruginosa response regulators in virulence. Moreover, the input signal that triggers the response regulator activation is yet to be uncovered for most systems. To find new response regulators involved in virulence, in vitro infections werecarried out using macrophages. Briefly, the macrophages were infected with each response regulator mutant or the wild-type strain, the pro-inflammatory cytokine production (TNF-α) and the bacterial clearance were evaluated. Using this approach, we identified several response regulators involved in virulence, and we also confirmed the involvement of known response regulators in this process. One of the novel virulence-related response regulators, PA14_26570 (named here as AtvR), is an atypical response regulator with a substitution in the phosphorylable aspartate to glutamate, that usually leads to an always-on state. A non-polar mutant was constructed, and macrophage infection with ΔatvR confirmed an increased bacterial clearance as well as a higher TNF-α production as compared to the wild-type strain. To ascertain the role of AtvR during the pathogenic process, an acute pneumonia model was used. Mice infected with ΔatvR showed an increased survival as compared to mice infected with the wildtype strain. In addition, ΔatvR infected mice showed reduced bacterial burden, increased neutrophil recruitment and activation, as well as increased pro-inflammatory cytokine production (TNF-α and IFN-γ). Also, using a transcriptomic approach (RNASeq), we showed that several genes were upregulated in the strain overexpressing AtvR. These genes include the anaerobic respiration clusters nar, nir, nor and nos. This result was confirmed by qRT-PCR and phenotypic analysis, in which ΔatvR showed reduced growth and nitrate reductase expression during hypoxic conditions as compared to the wild-type strain. In conclusion, we have demonstrated that several response regulators are important for P. aeruginosa virulence in vitro. In addition, we further characterized the atypical response regulator AtvR, which regulates anaerobic respiration via denitrification, allowing this bacterium to infect and colonize the host more efficiently

Complete genome sequence of the aerobically denitrifying thermophilic bacterium Chelatococcus daeguensis TAD1

ABSTRACT Chelatococcus daeguensis TAD1 is a themophilic bacterium isolated from a biotrickling filter used to treat NOx in Ruiming Power Plant, located in Guangzhou, China, which shows an excellent aerobic denitrification activity at high temperature. The complete genome sequence of this strain was reported in the present study. Genes related to the aerobic denitrification were identified through whole genome analysis. This work will facilitate the mechanism of aerobic denitrification and provide evidence for its potential application in the nitrogen removal.

El impacto de la hidroquinona usada como un modelo de efector redox sobre la desnitrificación potencial, la actividad microbiana y las condiciones redox de un suelo cultivable / Impact of hydroquinone used as a redox effector model on potential denitrification, microbial activity and redox condition of a cultivable soil

In this microcosm study, we analyzed the effect produced by hydroquinone on the expression of soil biological denitrification, in relation to the redox state of the soil, both in terms of intensity factor (Eh′) and capacity factor (amount of oxidized or reduced compounds). The supplementation of an Argiudoll soil with hydroquinone decreased the soil apparent reduction potential (Eh′) and soil dehydrogenase activity (formazan production from tetrazolium chloride reduction; redox capacity factor), the relationship between both factors being highly significative, r = 0.99 (p < 0.001). The bacterial population (measured by colony forming units) increased, and the production of N2O was greater (p < 0.001) at 200 and 400 μg/g dry soil doses. Furthermore, there was an inverse relationship between soil dehydrogenase activity and the number of bacteria (r = −0.82; p < 0.05), increased denitrification activity and changes in the CO2/N2O ratio value. These results suggest that hydroquinone at supplemented doses modified the soil redox state and the functional structure of the microbial population. Acetate supplementation on soil with hydroquinone, to ensure the availability of an energy source for microbial development, confirmed the tendency of the results obtained with the supplementation of hydroquinone alone. The differences observed at increased doses of hydroquinone might be explained by differences on the hydroquinone redox species between treatments.

En este trabajo estudiamos, en condiciones de microcosmos, el efecto que produce la hidroquinona sobre la expresión de la desnitrificación en relación con el estado de óxido-reducción del suelo, en términos de factor de intensidad (Eh′) y de factor de capacidad (cantidad de compuestos oxidados o reducidos). La suplementación de un suelo argiudol con hidroquinona disminuyó el potencial de reducción aparente (Eh′) y la actividad deshidrogenasa (producción de formazán a partir de la reducción de cloruro de tetrazolio; factor de capacidad redox), la relación entre ambos factores fue altamente significativa, r = 0,99 (p < 0,001). La población bacteriana heterotrófica (medida como unidades formadoras de colonias) aumentó y la producción de N2O fue mayor (p < 0,001) con las dosis de 200 y 400 μg/g de suelo seco. Además se observó una relación inversa entre la producción de formazán y el número de bacterias (r = −0,82; p < 0,05), la actividad desnitrificadora aumentó y se produjeron cambios en el valor del cociente CO2/N2O. Estos resultados sugieren que la hidroquinona, en las dosis empleadas, modificó el estado redox del suelo y la estructura funcional de la población microbiana. La suplementación con acetato en el suelo con hidroquinona, a fin de asegurar la disponibilidad de una fuente de energía para el desarrollo bacteriano, confirmó la tendencia de los resultados obtenidos con la suplementación con hidroquinona solamente. Las diferencias observadas con el incremento en la dosis de hidroquinona podrían explicarse por las diferencias sobre las especies redox de la hidroquinona entre los tratamientos.

Application and obstacles of ANAMMOX process / 生物工程学报

Anaerobic ammonium oxidation (ANAMMOX), as its essential advantages of high efficiency and low cost, is a promising novel biological nitrogen elimination process with attractive application prospects. Over the past two decades, many processes based on the ANAMMOX reaction have been continuously studied and applied to practical engineering, with the perspective of reaching 100 full-scale installations in operation worldwide by 2014. Our review summarizes various forms of ANAMMOX processes, including partial nitritation-ANAMMOX, completely autotrophic nitrogen removal over nitrite, oxygen limited autotrophic nitrification and denitrification, denitrifying ammonium oxidation, aerobic deammonification, simultaneous partial nitrification, ANAMMOX and denitrification, single-stage nitrogen removal using ANAMMOX and partial nitritation. We also compare the operating conditions for one-stage and two-stage processes and summarize the obstacles and countermeasures in engineering application of ANAMMOX systems, such as moving bed biofilm reactor, sequencing batch reactor and granular sludge reactor. Finally, we discuss the future research and application direction, which should focus on the optimization of operating conditions and applicability of the process to the actual wastewater, especially on automated control and the impact of special wastewater composition on process performance.

Desempeño de un reactor biológico secuencial (RBS) en el tratamiento de aguas residuales domésticas / Performance of a Sequencing Batch Reactor (SBR) in the treatment of domestic sewage

Se estudió la remoción biológica de materia orgánica y nutrientes de un agua residual doméstica empleando un Reactor Biológico Secuencial (RBS) a escala piloto. El estudio fue dividido en cuatro fases en las que se modifico la carga orgánica y la duración de las etapas anaerobia, aerobia y anoxica que conforman cada ciclo de tratamiento, considerando edades de lodo de 10 y 7,5 días. Durante las Fases I y II se operó el sistema con bajos valores de carga másica: 0,364 y 0,220 kg.DQO/Kg.SSV.dia, mientras que durante las Fases III y IV se emplearon cargas mayores: 0,665 y 0,737 kg.DQO/Kg.SSV.dia respectivamente. Los resultados obtenidos muestran que las mayores eficiencias de remoción de materia orgánica en términos de DBO se alcanzaron durante la Fases III (91%) y IV (82%), con remoción de fósforo superior a 40%. En cuanto al proceso de nitrificación durante las Fases I y II se registraron tasas de 0,032 y 0,024 kg.N-NH3/kg.SSV.dia, esto debido al menor contenido de materia orgánica y a la baja relación DBO/NKT, mientras que durante las Fases III y IV estas fueron menores: 0,015 kg.N-NH3/kg.SSV.dia durante la Fase III y 0,020 kg.N-NH3/kg.SSV.dia en la Fase IV, sin embargo, fue en estas fases donde se alcanzaron los mayores niveles de desnitrificación durante la etapa anóxica, favorecido por una relación C/N adecuada, próxima de 4 kg.DBO/kg.N-NO3- y la presencia de un substrato de fácil biodegradación. Los resultados obtenidos muestran los RBS como una alternativa eficiente y viable en el tratamiento de aguas residuales domésticas.

It was studied the biological removal of organic matter and nutrients from domestic wastewater using a Sequential Biological Reactor (SBR) at pilot scale. The study was divided into four phases in which the organic load and the duration of anaerobic, aerobic and anoxic cycle were modified, considering sludge ages of 10 and 7.5 days. During Phases I and II the system was operated with low mass load values: 0.364 and 0.220 kg.COD/kg.VSS.day, while during Phases III and IV were used heavier loads: 0.665 and 0.737 kg.COD/Kg.VSS.day respectively. The results obtained show that the removal efficiencies of organic matter in terms of BOD were achieved during Phase III (91%) and IV (82%) with phosphorus removal exceeding 40%. As for the nitrification process during Phases I and II showed rates of 0.032 and 0.024 kg.N-NH3/kg.VSS.day, this due to lower organic matter content and low BOD/TKN, while during phases III and IV these were lower: 0.015 kg.N-NH3/kg.VSS.day during Phase III and 0.020 kg.N-NH3/kg.VSS.day in Phase IV, however, was in these phases which reached the highest levels of denitrification during the anoxic phase, favored by a C/N appropriate next of 4 kg.BOD/kg.N-NO3- and the presence of an easily biodegradable substrate. The results show the RBS as an efficient and viable process in the treatment of domestic wastewater.

Nitrificação e desnitrificação simultânea em reator com biomassa em suspensão e fluxo contínuo de esgoto / Simultaneous nitrification and denitrification in reactor with biomass in suspension and continuous flow of sewage

Recentemente foi desenvolvido na Europa o processo conhecido por Nitrificação e Desnitrificação Simultânea (NDS), em que o lodo ativado opera com tempo de retenção de sólidos suficientemente altos para que a nitrificação ocorra mesmo com baixa concentração de oxigênio dissolvido nos tanques de aeração, situação em que é possível a manutenção de condições anóxicas no interior dos flocos e a ocorrência da desnitrificação. Em países de clima tropical, muitas estações de tratamento de esgoto foram construídas ou estão sendo projetadas para operarem sob as condições da variante do processo de lodo ativado conhecida por aeração prolongada, reconhecidamente eficiente na remoção de matéria orgânica e na nitrificação do esgoto. Tais estações podem ser adaptadas para estas novas condições, de forma a se obter o benefício técnico da desnitrificação e o ganho econômico relativo à redução no consumo de energia elétrica para aeração. Este estudo avaliou as condições de remoção de nitrogênio via NDS em um sistema de lodo ativado com aeração prolongada alimentada em fluxo contínuo com diferentes idades do lodo 40, 30 e 20 dias, tratando esgoto sanitário. O processo de tratamento permaneceu estável durante todo o período de observação. Os resultados experimentais demonstram que é possível manter baixas concentrações de OD no tanque de aeração em uma faixa relativamente estreita (0,3-0,8 mg.L-1), e, consequentemente desenvolver os processos de nitrificação e desnitrificação sob as mesmas condições (NDS) com alta eficiência e estabilidade. A remoção de DQO e DBO foi acima de 90 por cento . Os valores de crescimento específico máximo (m20) das bactérias autotróficas nitritantes e nitratantes, resultaram muito próximos aos descritos na literatura, sendo para a idade do lodo de 20 dias os valores de 0,68 d-1 para as bactérias nitritantes e de 0,62 d-1 para as bactérias nitratantes. Os valores das taxas de utilização máxima de amônia foram maiores do que as obtidas com o nitrito e, pode-se observar que o acúmulo de nitrato devido à diminuição da amônia foi mais rápido do que o com nitrito. Desse modo, conclui-se que a nitrificação foi governada pelas bactérias autotróficas nitritantes. A taxa de utilização de nitrato foi maior tendo como substrato material rapidamente biodegradável. Confirmando que a disponibilidade de carbono tem papel fundamental nas taxas de desnitrificação no processo de NDS.

investigation on the nitrogen content of a petroleum refinery's wastewater and its removal by biological treatment

This investigation was performed on the biological removal of nitrogen from refinery's wastewater by the nitrification and denitrification process. In a petroleum refinery, removing of hydrocarbons is the main concern and nitrogen content is supposed to be negligible. The aim of this work was to search for nitrogen in Tehran Refinery wastewater and employing a biological technology to reduce this pollutant. Samplings were done in different points of wastewater treatment plant; influent to aeration unit, effluent of aeration unit and effluent of clarifiers. The results showed that despite of a high average removal efficiency of COD > 93%, the nitrogen removal during conventional activated sludge process was not efficient and sludge rising due to denitrification was observed within the clarifier. The analysis conducted in laboratory scale showed that a simultaneous nitrification and denitrification [SND] process could easily be realized in the same activated sludge plant by using the flocculating sludge and control of dissolved oxygen concentration. It was found that the higher MLSS value [10.0 g/L] and mixing rate [300 rpm] is effective in improving total nitrogen removal and overall SND performance. Our experimental results indicated that the SND process is very efficient for nitrogen removal from industrial wastewater

Isolation and characterization of a novel denitrifying bacterium with high nitrate removal: Pseudomonas stutzeri

The aim of this study was to isolate and characterize a high efficiency denitrifier bacterium for reducing nitrate in wastewater. Six denitrifier bacteria with nitrate removal activities were isolated from a petrochemical industry effluent with high salinity and high nitrogen concentrations without treatment. The isolated bacteria were tested for nitrate reomoval activity. One of the bacterium displayed the highest reduction of nitrate. The strain was preliminarily identified using biochemical tests and further identified based on similarity of PCR-16S rRNA using universal primers. Biochemical and molecular experiments showed that the best bacterium with high nitrate removal potential was Pseudomonas stutzeri, a member of the alpha subclass of the class Proteobacteria. The extent of nitrate removal efficiency was 99% at 200 mg/L NO[3] and the nitrite content of the effluent was in the prescribed limit. The experiments showed the ability of Pseudomonas stutzeri to rapidly remove nitrate under anoxic conditions. The strain showed to be potentially good candidate for biodenitrification of high nitrate solutions

Performance of moving bed biofilm reactors for biological nitrogen compounds removal from wastewater by partial nitrification-denitrification process

In this research, the continuously operated laboratory scale Kaldnes [k1] moving bed biofilm reactors [MBBRs] under partial nitrification-denitrification process were used for treatment of synthetic wastewater containing ammonium and glucose. The Anoxic and Aerobic reactors were filled to 40 and 50% [v/v] to attach and retain biomass with k1 biofilm carriers, respectively. The favorite internal recycle ratio and hydraulic residence time [HRT] to eliminate nitrogen compounds were 300% of inflow rate and 20 hours, respectively. Optimal dissolved oxygen [DO] was 1-1.5 mg/L in the aerobic reactor. No sludge was returned into the system and only an internal recycling was performed from aerobic to anoxic reactor. The results showed that the maximum and average specific nitrification rate [SNR] in the aerobic reactor were 49.4 and 16.6 g NOx-N/KgVSS.day, respectively and the maximum and average specific denitrification rate [SDNR] as 156.8 and 40.1gNOx-N/KgVSS.day in the anoxic reactor, respectively. The results also showed that it is possible to reach a stable partial nitrification with high ratio of NO2-N/NOx-N [80% to 85%] during high load ammonium and low DO concentration [<1.5 mg/L] in the aerobic reactor. During optimum conditions, the average removal efficiency of total nitrogen [TN], ammonia and soluble organic carbon [SCOD] occurred as 98.23%, 99.75% and 99.4%, respectively. This study showed that the partial nitrification/denitrification process in the moving bed biofilm reactors system has an acceptable performance for treatment of wastewater with high load of organic carbon and organic nitrogen compounds

Influence of metal ions and organic carbons on denitrification activity of the halotolerant bacterium, Paracoccus pantotrophus P16 a strain from shrimp pond

The effect of metal ions, ferric ion (Fe3+) and molybdenum ion (Mo6+) on the denitrification process of Paracoccus pantotrophus P16 grown under saline conditions was investigated. Results revealed that the dosages of added Fe3+ and Mo6+ significantly accelerated nitrate utilization and nitrite accumulation. Enzymatic studies revealed that the membrane-bound nitrate reductase and the periplasmic nitrite reductase had activities of 998 +/- 28 and 373 +/- 18 nmol (mg protein)-1 min-1, respectively after growing Paracoccus pantotrophus P16 in medium supplemented with 1.5 micron M Fe3+. If provided with 1.5 micron M Fe3+and 2.4 micron M Mo6+, the membrane-bound nitrate reductase activity increased to 6,223 +/- 502 nmol (mg protein)-1 min-1 and the periplasmic nitrite reductase was 344 +/- 20 nmol (mg protein)-1 min-1. The results indicated that an addition of Fe3+ and Mo6+ led to an overstimulation of nitrate reductase activity as compared with nitrite reductase activity. When glucose was supplied, the minimal ratio of carbon per nitrate (C/N) was 2.31 mg C/mg NO3--N with denitrification yield of 0.45 g NO3--N/g C. Addition of ethanol instead of glucose, the minimal ratio of C/N was 1.15 mg C/mg NO3--N with denitrification yield of 1.08 g NO3--N/g C.