• 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.

• Journal of Microbiology and Biotechnology
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• 제19권12호
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• pp.1656-1664
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• 2009

Bioaugmentation of bioreactors focuses on the removal of numerous organics, with little attention typically paid to the maintenance of high and stable nitrite accumulation in partial nitrification. In this study, a bioaugmented membrane bioreactor (MBR) inoculated with enriched ammonia-oxidizing bacteria (AOB) was developed, and the effects of dissolved oxygen (DO) and temperature on the stability of partial nitrification and microbial community structure, in particular on the nitrifying community, were evaluated. The results showed that DO and temperature played the most important roles in the stability of partial nitrification in the bioaugmented MBR. The optimal operation conditions were found at 2-3 mgDO/l and $30^{\circ}C$, achieving 95% ammonia oxidization efficiency and nitrite ratio ($NO_2^-/{NO_x}^-$) of 0.95. High DO (5-6 mg/l) and low temperature ($20^{\circ}C$) had negative impacts on nitrite accumulation, leading to nitrite ratio drop to 0.6. However, the nitrite ratio achieved in the bioaugmented MBR was higher than that in most previous literatures. Denaturing gradient gel electrophoresis (DGGE) and fluorescence in situ hybridization (FISH) were used to provide an insight into the microbial community. It showed that Nitrosomonas-like species as the only detected AOB remained predominant in the bioaugmented MBR all the time, and coexisted with numerous heterotrophic bacteria. The heterotrophic bacteria responsible for mineralizing soluble microbial products (SMP) produced by nitrifiers belonged to the Cytophaga-Flavobacterium-Bacteroides (CFB) group, and $\alpha$-, $\beta$-, and $\gamma$- Proteobacteria. The fraction of AOB ranging from 77% to 54% was much higher than that of nitrite-oxidizing bacteria (0.4-0.9%), which might be the primary cause for the high and stable nitrite accumulation in the bioaugmented MBR.

• 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.

• 대한환경공학회지
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• 제28권10호
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• pp.1024-1030
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• 2006

Genus Nitrospira와 Nitrobacter는 폐수 질산화 시스템의 대표적인 아질산 산화균으로 알려져 있다. Genus Nitrospira는 아질산 농도가 매우 낮은 조건에서도 이를 효율적으로 활용하는 K-strategists로 알려져 있는 반면에 Nitrobacter 종은 기질소비와 성장이 빠른 r-strategists로 알려져 왔다. 또한 유기물이나 용존산소도 아질산 산화균의 분포에 영향을 주는 것으로 알려져 있다. 본 연구에서는 아질산과 유기물, 용존산소가 복합적으로 작용하는 질산화 시스템에서 아질산 산화균의 분포와 경쟁에 어떻게 영향을 받는지를 검토하였다. 이를 위하여 실험실 규모의 질산화 생물반응기와 질산화-탈질을 수행하는 $A_2O$ 계열의 실제 폐수처리장에서 여러 항목과 두 종류의 아질산 산화균 분포를 측정, 비교하였다. 그 결과 아질산 농도는 평균 5 mg-N/L로 유지되며, 호기조건에서 유기물이 매우 낮게 유지되는 실험실 질산화 생물반응기는 Nitrobacter가, 호기-무산소 조건에서 질산화-탈질이 일어나고 아질산이 거의 없는 상태이며 유기물이 비교적 높게 유지되는 $A_2O$ 폐수처리장은 Nitrospira가 우점종으로 분포하였다. 이것은 여러 인자가 복합적으로 작용하는 상태에서는 아질산 산화균의 분포가 유기물과 용존산소 보다는 아질산 농도가 가장 중요한 인자임을 보여주며 기질 친화도가 낮지만 반응속도가 빠른 Nitrobacter가 r-strategist, genus Nitrospira는 기질친화도가 높은 K-strategist인 특성을 보임을 확인하였다.

• Journal of Microbiology and Biotechnology
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• 제18권8호
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• pp.1470-1474
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• 2008

Nitrospira is a dominant member of nitrite-oxidizing bacteria (NOB) in nitrifying bioreactors as well as in natural habitats. In this study, Nitrospira NOB were investigated in the two nitrifying reactors operated with high and low dissolved oxygen (DO) concentrations for a period of 300 days. Phylogenetic and terminal restriction fragment length polymorphism analyses based on 168 rRNA gene sequences revealed that the Nitrospira community compositions of the two reactors during the early period related to group 1 and half of the Nitrospira community composition shifted to group 2 in the high-DO reactor after day 179, although there was no significant change in the low-DO reactor. These results suggested that DO was an important factor affecting Nitrospira community compositions in the nitrifying reactors.

• 한국미생물·생명공학회지
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• 제47권2호
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• pp.296-302
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• 2019

In this study, the effects of ammonium concentration ($117.5-1155.0mg-N{\cdot}l^{-1}$), nitrite concentration ($0-50.0mg-N{\cdot}l^{-1}$), and temperature ($15-35^{\circ}C$) on nitrification performance and its functional genes (amoA-arc, amoA-bac, hao) in an enriched consortium inoculated with humic acid were determined. Notably, the maximum nitrification rate value was observed at $315mg-N{\cdot}l^{-1}$ of ammonium, but the highest functional gene copy numbers were obtained at $630mg-N{\cdot}l^{-1}$ of ammonium. No inhibition of the nitrification rate and functional gene copy numbers was observed via the added nitrites. The optimum temperature for maximum nitrification performance was observed to be $30^{\circ}C$. The amoA-bac copy numbers were also greater than those of amoA-arc under all test conditions. Notably, amoA-arc copy numbers and nitrification efficiency showed a positive relationship in network analysis. These results indicate that ammonium-oxidizing archaea and bacteria play important roles in the nitrification process.

• 생태와환경
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• 제37권4호통권109호
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• pp.363-367
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• 2004

낙동강 전 수계 8개 정점에서 분자기법인 FISH (Fluorescent in situ Hybridization)법으로 세균군집구조를 비교분석하였다. Eubacteria에 ${\alpha}\;{\cdot}\;{\beta}\;{\cdot}\;{\gamma}-subclasses$ proteobacter CF group 세균의 합이 총세군수에서 차지하는 비율이 9.3-42.5%에서 변화하였고 그 최고치는 히상류, 청량에서 나타났다. 각 세균그룹이 총세균수에서 차지하는 비율이 10% 미만이었으나 최상류에서의 CF그룹이 총세균수에서 차지하는 비율은 23%이었다. 또한 유기물질을 분해해서 빠른 성장을 한다는 ${\gamma}-subclasses$ proteobacteria 세균군이 예상과는 달리 유기오염정도가 높은 하류에 비해 상류에서 더 많이 검출되었다. 아울러 암모니아산화세균은 $2.7-18.0{\times}10^4$ cells $mL^{-1}$의 범위에서 변화하였고 하류에서 최저치를 그리고 최고치는 중류에서 보였다. 반면에 아질산산화세균의 경우, 전수계에 걸쳐 정점간의 별 차이 없이 $5.2-7.7{\times}10^4$ Cells $mL^{-1}$에서 변화하였으며 그들이 총세균수에서 차지하는 비율은 두세균군간의 차이없이 1.0-13.6%에서 변화하였다. 결론적으로 FISH법은 통상적으로 세균군집의 정량적인 분석에 사용되지만 그 결과는 본 연구결과에서 보는 바와 같이 수계 환경의 현황에 관한 좋은 정보를 제공해주기도 한다.

• 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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• 제22권6호
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• pp.1014-1021
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• 2006

SMMIAR (Submerged Moving Media Intermittent Aeration Reactor) Process is a very efficient system which remove ammonia to nitrogen gas in one reactor. In this study, we determined the diversity of ammonia oxidizing bacteria and denitrifying bacteria using specific PCR amplification and the clone library construction. An ammonia monooxygenase gene(amoA) was analyzed to investigate the diversity of nitrifiers. Most of amoA gene fragments (27/29, 93%) were same types and they are very similar (>99%) to the sequences of Nitrosomonas europaea and other clones isolated from anoxic ammonia oxidizing reactors. ANAMMOX related bacteria have not determined by specific PCR amplification. A nitrite reductase gene(nirK) was analyzed to investigate the diversity of denitrifying bacteria. About half (9/20, 45%) of denitrifiers were clustered with Rhodobacter and most of others were clustered with Mesorhizobium (6/20, 30%) and Rhizobium (3/20, 15%). All of these nirK gene clones were clustered in alpha-Proteobacteria and this result is coincide with other system which also operate nitrification and denitrification in one reactor. The molecular monitoring of the population of nitrifiers and denitrifiers would be helpful for the system stabilization and scale-up.

• 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.