• 한국물환경학회지
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• 제36권3호
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• pp.220-228
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• 2020

Anaerobic ammonium oxidation (AMX) is a cost-efficient biological nitrogen removal process. The coexistence of ammonia-oxidizing bacteria (AOB) in an AMX reactor is an interesting research topic as a nitrogen-related bacterial consortium. In this study, a sequencing batch reactor for AMX (AMX-SBR) was operated with a conventional activated sludge. The AOB in an AMX bioreactor were identified and quantified using terminal restriction fragment length polymorphism (T-RFLP) and real-time qPCR. A T-RFLP assay based on the ammonia monooxygenase subunit A (amoA) gene sequences showed the presence of Nitrosomonas europaea-like AOB in the AMX-SBR. A phylogenetic tree based on the sequenced amoA gene showed that AOB were affiliated with the Nitrosomonas europaea/mobilis cluster. Throughout the enrichment period, the AOB population was stable with predominant Nitrosomonas europaea-like AOB. Two OTUs of amoA_SBR_JJY_20 (FJ577843) and amoA_SBR_JJY_9 (FJ577849) are similar to the clones from AMX-related environments. Real-time qPCR was used to quantify AOB populations over time. Interestingly, the exponential growth of AOB populations was observed during the substrate inhibition of the AMX bacteria. The specific growth rate of AOB under anaerobic conditions was only 0.111 d^-1. The growth property of Nitrosomonas europaea-like AOB may provide fundamental information about the metabolic relationship between the AMX bacteria and AOB.

• 한국환경농학회지
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• 제39권1호
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• pp.26-35
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• 2020

BACKGROUND: Microbes that govern a unique biochemical process of oxidizing ammonia into dinitrogen gas, such as anaerobic ammonium oxidation (anammox) have been reported to play a pivotal role in agricultural soils and in oceanic environments. However, limited information for anammox bacterial abundance and distribution in the terrestrial habitats has been known. METHODS AND RESULTS: Phylogenetic and next-generation sequencing analyses of bacterial 16S rRNA gene were performed to examine potential anammox bacteria in paddy soils. Through clone libraries constructed by using the anammox bacteria-specific primers, some clones showed sequence similarities with Planctomycetes (87% to 99%) and anammox bacteria (94% to 95%). Microbial community analysis for the paddy soils by using Illumina Miseq sequencing of 16S rRNA gene at phylum level was dominated by unclassified Bacteria at 33.2 ± 7.6%, followed by Chloroflexi at 20.4 ± 2.0% and Acidobacteria at 17.0 ± 6.5%. Planctomycetes that anammox bacteria are belonged to was 1.5% (± 0.3) on average from the two paddy soils. CONCLUSION: We suggest evidence of anammox bacteria in the paddy soil. In addition to the relatively well-known microbial processes for nitrogen-cycle, anammox can be a potential contributor on the cycle in terrestrial environments such as paddy soils.

• 상하수도학회지
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• 제32권5호
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• pp.371-379
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• 2018

The lack of seed sludges for Ammonium Oxidizing Bacteria (AOB) and slow-growing ANaerobic AMMonium OXidation (ANAMMOX) bacteria is one of the major problem for large-scale application. In this study, $24m^3$ of single-stage SBR (Sequencing Batch Reactor) was operated to remove nitrogen from reject water using AOB and ANAMMOX bacteria cultivated from activated sludge in the field. The ANAMMOX activity was found after 44 days of cultivation in the ANAMMOX cultivation reactor, and then $0.66kg\;N/m^3/d$ of the nitrogen removal rate was achieved at $0.78kg\;N/m^3/d$ of the nitrogen loading rate at 153 days of cultivation. The AOB cultivation reactor showed $0.2kg\;N/m^3/d$ of nitrite production rate at $0.4kg\;N/m^3/d$ of nitrogen loading rate after 36 days of operation. The cultivated ANAMMOX bacteria and AOB was mixed into the single-stage SBR. The feed distribution was applied to remove total nitrogen stably in the single-stage SBR. The nitrogen removal rate in the single-stage SBR was gradually enhanced with an increase of specific activities of both AOB and ANAMMOX bacteria by showing $0.49kg\;N/m^3/d$ of the nitrogen removal rate at $0.56kg\;N/m^3/d$ of the nitrogen loading rate at 54 days of operation.

• Journal of Microbiology and Biotechnology
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• 제27권7호
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• pp.1288-1299
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• 2017

Composting is widely used to transform waste into valuable agricultural organic fertilizer. Anaerobic ammonium-oxidizing (anammox) bacteria play an important role in the global nitrogen cycle, but their role in composting remains poorly understood. In the present study, the community structure, diversity, and abundance of anammox bacteria were analyzed using cloning and sequencing methods by targeting the 16S rRNA gene and the hydrazine oxidase gene (hzo) in samples isolated from compost produced from cow manure and rice straw. A total of 25 operational taxonomic units were classified based on 16S rRNA gene clone libraries, and 14 operational taxonomic units were classified based on hzo gene clone libraries. The phylogenetic tree analysis of the 16S rRNA gene and deduced HZO protein sequences from the corresponding encoding genes indicated that the majority of the obtained clones were related to the known anammox bacteria Candidatus "Brocadia," Candidatus "Kuenenia," and Candidatus "Scalindua." The abundances of anammox bacteria were determined by quantitative PCR, and between $2.13{\times}10^5$ and $1.15{\times}10^6$ 16S rRNA gene copies per gram of compost were found. This study provides the first demonstration of the existence of anammox bacteria with limited diversity in cow manure composting.

• 한국물환경학회지
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• 제29권3호
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• pp.377-382
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• 2013

Enriching anammox bacteria (AMX) in a lab-scale granular sequencing batch reactor using local digester centrate, we observed the significant enrichment of the filamentous-like bacterial population. These bacteria were revealed as novel bacterial species (termed CHL) belonging to Chlorobi/Bacteroidetes phyla via Denaturing Gradient Gel Electrophoresis (DGGE). Further, niche differentiation of AMX and CHL quantification was observed in granule and filament biomass, suggesting AMX was dominant in the granule and CHL was dominant in the filament. Therefore, it was confirmed the structural role of CHL was indeed to aid the granule formation of the AMX. In parallel, the physiological role of CHL was suspected to degrade biopolymers in the digester centrate using nitrate as an electron acceptor.

• Journal of Microbiology and Biotechnology
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• 제22권7호
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• pp.1005-1014
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• 2012

The understanding of the relationship between ammonia-oxidizing bacteria (AOB) communities in activated sludge and the operational treatment parameters supports the control of the treatment of ammonia-rich wastewater. The modifications of treatment parameters by alteration of the number and length of aerobic and anaerobic stages in the sequencing batch reactor (SBR) working cycle may influence the efficiency of ammonium oxidation and induce changes in the AOB community. Therefore, in the research, the impact of an SBR cycle mode with alternating aeration/mixing conditions (7 h/1 h vs. 4 h/5.5 h) and volumetric exchange rate (n) on AOB abundance and diversity in activated sludge during the treatment of anaerobic sludge digester supernatant at limited oxygen concentration in the aeration stage (0.7 mg $O_2/l$) was assessed. AOB diversity expressed by the Shannon-Wiener index (H') was determined by the cycle mode. At aeration/mixing stage lengths of 7 h/1 h, H' averaged $2.48{\pm}0.17$, while at 4 h/5.5 h it was $2.35{\pm}0.16$. At the given mode, AOB diversity decreased with increasing n. The cycle mode did not affect AOB abundance; however, a higher AOB abundance in activated sludge was promoted by decreasing the volumetric exchange rate. The sequences clustering with Nitrosospira sp. NpAV revealed the uniqueness of the AOB community and the simultaneously lower ability of adaptation of Nitrosospira sp. to the operational parameters applied in comparison with Nitrosomonas sp.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2005년도 International Meeting of the Microbiological Society of Korea
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• pp.169-169
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• 2005

• 상하수도학회지
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• 제28권2호
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• pp.195-206
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• 2014

Immobilization of anaerobic ammonium oxidizing bacteria has been studied to enhance the biomass retention of the slowly growing bacteria and the process stability. The purpose of this study was to compare the nitrogen removal efficiency of granular and immobilized anammox bacteria with poly vinyl alcohol and alginate. The specific anammox activity of the granular, homoginized and immobilized anammox bacteria were $0.016{\pm}0.0002gN/gVSS/d$, $0.011{\pm}0.001gN/gVSS/d$ and $0.007{\pm}0.0005gN/gVSS/d$, respectively. Although the activity decreased to 43.7 % of the original one due to low pH and $O_2$ exposure during the homogination and the immobilization, it was rapidly recovered within 7 days in the following continuous culture. When synthetic T-N concentrations of 100, 200, 400, 800 mg/L were fed, the immobilized anammox bacteria showed higher nitrogen removal efficiencies at all operational conditions than those of granular anammox bacteria. When the sludge retention time was shorten below 30.7 days and the reject water was fed, the nitrite removal efficiency of the granular anammox bacteria dropped to 8 % of the initial value, while that of the immobilized anammox bacteria was maintained over 95 % of the initial one. The immobilization with poly vinyl alcohol and alginate would be a feasible method to improve the performance and stability of the anammox process.

• Journal of Microbiology and Biotechnology
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• 제27권10호
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• pp.1798-1807
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• 2017

The differentiations in nitrogen-converting activity and microbial community structure between granular size fractions in a continuous completely autotrophic nitrogen removal over nitrite (CANON) reactor, having a superior specific nitrogen removal rate of $0.24g/(g\;VSS{\cdot}h)$, were investigated by batch tests and high-throughput pyrosequencing analysis, respectively. Results revealed that a high dissolved oxygen concentration (>1.8 mg/l) could result in efficient nitrite accumulation with small granules (0.2-0.6 mm in diameter), because aerobic ammonium-oxidizing bacteria (genus Nitrosomonas) predominated therein. Meanwhile, intermediate size granules (1.4-2.0 mm in diameter) showed the highest nitrogen removal activity of $40.4mg/(g\;VSS{\cdot}h)$ under sufficient oxygen supply, corresponding to the relative abundance ratio of aerobic to anaerobic ammonium-oxidizing bacteria (genus Candidatus Kuenenia) of 5.7. Additionally, a dual substrate competition for oxygen and nitrite would be considered as the main mechanism for repression of nitrite-oxidizing bacteria, and the few Nitrospira spp. did not remarkably affect the overall performance of the reactor. Because all the granular size fractions could accomplish the CANON process independently under oxygen limiting conditions, maintaining a diversity of granular size would facilitate the stability of the suspended growth CANON system.

• 한국물환경학회지
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• 제38권4호
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• pp.171-176
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• 2022

Recently, an autotrophic single-stage nitrogen removal (ASSNR) process based on the anaerobic ammonium oxidation (ANAMMOX) reaction has been proven as an economical ammonia treatment. It is highly evident that double-layered gel beads are a promising alternative to the natural biofilm for ASSNR because of the high mechanical strength of poly(vinyl alcohol) (PVA)/alginate structure and efficient protection of ANAMMOX bacteria from dissolved oxygen (DO) due to the thick outer layer. However, the thick outer layer results in severe mass transport limitation and consequent lowered bacterial activity. Therefore, the effects of the thickness of the outer layer on the overall reaction rate were tested in the biofilm model using AQUASIM for ammonia-oxidizing bacteria (AOB), nitrite-oxidizing bacteria (NOB) and ANAMMOX bacteria. A thickness of 0.5~1.0 mm is preferred for the maximum total nitrogen (TN) removal. In addition, a DO of 0.5 mg/L resulted in the best total nitrogen removal. A higher DO induces NOB activity and consequent lower TN removal efficiency. The optimal density of AO B and NO B density was 1~10% for a 10% ANAMMOX bacterial in the double-layered PVA/alginate gel beads. The real effects of operating parameters of the thickness of the outer layer, DO and concentrations of biomass balance should be intensively investigated in the controlled experiments in batch and continuous modes.