• 한국지하수토양환경학회지:지하수토양환경
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• 제21권4호
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• pp.50-59
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• 2016

Spatial and temporal changes in nitrification activities and distribution of microbial population of ammonia oxidizing bacteria (AOB) and ammonia oxidizing archaea (AOA) in paddy soils were investigated. Soil samples were collected in March and October 2015 from rice paddy with and without the presence of confined animal feeding operations. Incubation experiments and quantitative polymerase chain reaction showed that AOA's contribution to nitrification kinetics was much higher in locations where organic nitrogen in animal waste is expected to significantly contribute to overall nitrogen budget, and temporal variations in nitrification kinetics were much smaller for AOA than AOB. These differences were interpreted to indicate that different microbial responses of two microbial populations to the types and concentrations of nitrogen substrates were the main determining factors of nitrification processes in the paddy soils. The copy numbers of ammonium monooxygenase gene showed that AOA colonized the paddy soils in higher numbers than AOB with stable distribution while AOB showed variation especially in March. Although small in numbers, AOB population turned out to exert more influence on nitrification potential than AOA, which was attributed to higher fluctuation in AOB cell numbers and nitrification reaction rate per cells.

• 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.

• 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.

• Applied Biological Chemistry
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• 제46권1호
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• pp.1-6
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• 2003

본 연구는 폐수 중의 질소 제거를 위한 생물학적 처리용 미생물 개발을 위한 목적으로 질소의 산화 능력이 뛰어난 균주를 분리하였다. 분리된 세균 중에서 질소 산화능과 생육 속도가 뛰어난 CH-N 균주를 선별하였으며, 생리, 생화학적 특성 조사에 의해 Bacillus sp로 추정되어 Bacillus sp. CH-N이라 명명하였다. 분리 균주는 0.5% glucose가 포함된 초기 pH가 7,0인 암모니아 및 아질산성 질소 함유 배지에서 30시간 배양 후 각각 85%와 90%의 암모니아성과 아질산성 질소의 감소율을 나타내었다. 폐수 및 생활하수에 분리 균주를 이용한 결과, 수질 속의 암모니아성 질소가 단시간에 크게 감소시키는 효과를 확인하였다. 균주를 고정시킨 담체의 질소산화 효과를 시험하고자 Bacillus sp. CH-N을 고정시킨 세라믹 담체를 이용한 결과, 배양 2일 후에는 암모니아성 질소가 전부 제거되었다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제7권5호
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• pp.323-326
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• 2002

To detect whole ammonia-oxidizing bacteria in the activated sludge, group-specific primers targeting the 16S-rRNA gene of ammonia-oxidizing bacteria were used. The electrophoresis pattern of the PCR products seemed to produce a single band of approximately 1.0 k bp for the bacteria in activated sludge and Nitrosomonas europaea. No band was observed for nitrite-oxidizer Nitrobacter winogradskyi and heterotrophs such as Pseudomonas putida. Then direct measurement of the PCR product was made by fluorometry using the reagent Hoechist 33258, so that the fluorescent intensity was in proportional to the cell number of the sample up to 240. Total time required for the test was about 4 h including DNA extraction. The DNA fragments produced were cloned and their sequences showed high similarity to those of Nitrosomonas spp. This study showed the feasibility to detect ammonia-oxidizing bacteria and to esti-mate their population rapidly for the control of the nitrogen elimination process.

• 한국물환경학회지
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• 제24권3호
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• pp.383-390
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• 2008

Recently, the interests on economical nitrogen removal from wastewater are growing. As a method of the novel nitrogen removal technology, nitrogen removal via nitrite pathway by selective inhibition of free ammonia and free nitrous acid on nitrite oxidizing bacteria have been intensively studied. The inhibition effects of free ammonia and free nitrous acid are low when domestic wastewater is used, however, because of its relatively lower nitrogen concentration than the wastewater from industry and landfill, etc. In this study, a sequencing batch reactor (SBR) is proposed for nitrogen removal to investigate the effect of the low nitrogen concentration on nitrite accumulation. Nitrification efficiency reached almost 100% during the aerobic cycle and the maximum specific nitrification rate ($V_{max,nit}$) reached $17.8mg\;NH_4{^+}-N/g\;MLVSS{\bullet}h$. During the anoxic cycle, average denitrification efficiency reached 87% and the maximum specific denitrification rate ($V_{max,den}$) reached $9.8mg\;NO_3{^-}-N/g\;MLVSS{\bullet}h$. From the analysis the main reason of nitrite accumulation in the SBR was free nitrous acid rather than free ammonia. Nitrite accumulation increased with the decrease of organic content in the wastewater and the mechanism is not well understood yet. From the result of fluorescent in situ hybridization, the distribution of nitrite oxidizing bacteria was in equilibrium with ammonium oxidizing bacteria when nitrite accumulation did not occur.

• 미생물학회지
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• 제44권1호
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• pp.29-36
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• 2008

질소는 하수처리과정에서 제거되어야 하는 주요 오염물질 중의 하나이며, 세균 군집을 이용한 고도처리 시스템에서 생물학적 질소제거는 중요한 기술이다. 질산화반응은 생물학적 질소제거 시스템의 첫 단계로 미생물에 의해 진행된다. 암모니아는 암모니아산화세균에 의해 아질산염으로 산화되며, 그 후에 아질산염은 아질산염 산화세균에 의해 질산염으로 산화된다. 실험실 규모의 생물학적 질소제거 시스템인 변형된 eBAF 시스템, Nutrient removal laboratory 시스템과 반추기법을 적용한 rSBR 시스템의 질산화반응조 시료에서 16S rRNA 유전자를 이용한 terminal restriction fragment length polymorphism (T-RFLP) 방법으로 아질산염 산화세균군집을 분석하였다. 제한효소로 형성된 단편의 클러스터분석에서 Nitrobacter 군집은 각각의 폐수처리 시스템에 따라 군집의 차이가 있음이 나타났다. 그러나 Nitrospira 군집의 클러스터분석에서는 액체와 담체의 서식지 환경 차이에 의해 군집이 구분되었다.

• 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.

• 한국환경농학회지
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• 제33권4호
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• pp.314-320
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• 2014

BACKGROUND: Run off from agricultural sites contaminates water bodies with nitrogen which is toxic and causes eutrophication when excessively accumulated. Hence, the interest in monitoring nitrogen toxicity in aquatic environment has been continuously increasing. METHODS AND RESULTS: To detect a real time toxicity of various nitrogen compounds, we applied biomonitoring method (biosensor) based on sulfur-oxidizing bacteria (SOB). The toxicity biomonitoring test was conducted in semi-continuous mode in a reactor filled with sulfur particles (2~4 mm diameter) under aerobic condition. Relative toxicity was simply determined by measuring the change in electrical conductivity (EC). Various nitrogenous compounds at different concentrations were evaluated as a potential toxic substance. Nitrite was found to be very toxic to SOB with a 90% inhibition even when the concentration as low as 3 mg/L. However, nitrate and ammonia have any inhibitory effect on SOB's activity. CONCLUSION: The biosensor based on SOB responded sensitively to nitrite even at substantially low concentrations. Therefore, it can be used as a reliable biological alarm system for rapid detection of contaminants due to its simplicity and sensitive nature.

• 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.