• 한국물환경학회지
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• 제22권2호
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• pp.300-307
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• 2006

The combined SHARON (Single reactor system for High ammonium Removal Over Nitrite)-ANAMMOX (Anaerobic ammonium oxidation) reactor were operated in mesophilic condition ($35^{\circ}C$). In this study, microbial granulation and characteristics of SHARON and ANAMMOX sludges were investigated using settling test, Scanning Electron Microscopy (SEM) and Fluorescence In Situ Hybridization (FISH). In SHARON reactor, Aerobic granulation with diameter of 1.5~2.5 mm was accomplished but aerobic granulation was weaker than anaerobic granular sludge. Initial seed sludge of ANAMMOX reactor was used as attached media for biofilm growth. ANAMMOX sludge was more compact and rounder rather than seed sludge. Though ANAMMOX sludge has high activity, it has lower settling ability than the seed granule. The color of ANAMMOX sludge was changed from dark to redish brown granular with diameter of 1~2 mm. In FISH of ANAMMOX sludge, high fraction of Candidatus B. stuttgartiensis which paid great role of nitrogen conversion was detected. Also, FISH results reveals that ANAMMOX bacteria inhabit at inner parts near surface, having advantages in utilization of substrates and protection from oxygen inhibition.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권3호
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• pp.199-204
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• 2006

The purpose of this work was to evaluate the development of the anammox process by the use of granular sludge selected from a digestion reactor as a potential seed source in a lab-scale UASB (upflow anaerobic sludge blanket) reactor system. The reactor was operated for approximately 11 months and was fed by synthetic wastewater. After 200 days of feeding with $NH_4^+\;and\;NO_2^-$ as the main substrates, the biomass showed steady signs of ammonium consumption, resulting in over 60% of ammonium nitrogen removal. This report aims to present the results and to more closely examine what occurs after the onset of anammox activity, while the previous work described the start-up experiment and the presence of anammox bacteria in the enriched community using the fluorescence in situ hybridization (FISH) technique. By the last month of operation, the consumed $NO_2^--N/NH_4^+-N$ ratio in the UASB reactor was close to 1.32, the stoichiometric ratio of the anammox reaction. The obtained results from the influent-shutdown test suggested that nitrite concentration would be one key parameter that promotes the anammox reaction during the start-up enrichment of anammox bacteria from granular sludge. During the study period, the sludge color gradually changed from black to red-brownish.

• 상하수도학회지
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• 제26권3호
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• pp.399-408
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• 2012

The overall goal of this study was to characterize and quantify ammonia-oxidizing bacteria (AOB) in four different full-scale sequence batch reactor (SBR) wastewater treatment plants. Also, this study focused on assessing the occurrence of the alternative ammonia-oxidizing microbes such as anammox (anaerobic ammonia oxidation) bacteria (AMX) and ammonia-oxidizing archaea (AOA) in these systems. Based on total AOB numbers and the estimated cell density in the mixed liquor samples, AOB constituted 0.3 - 1.8% of the total bacterial population in the four WWTPs. Based on clone library, Nitrosomonas ureae-like AOB were dominant in plant A and B, while plant C and D had Nitrosomonas nitrosa-like AOB as major AOB group. The four different AMX primer sets targeting AMX 16S rRNA gene produced PCR amplicons distantly related to Chlamydia and Planctomycetales group bacteria. However, it was not clear these groups of bacteria perform anammox reaction in the SBR plants. Also, molecular evidence of AOA was found in one of the SBR plants, with a sequence located in the deep branch of the sediment creanarchaeota group.

• 한국물환경학회지
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• 제21권5호
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• pp.477-483
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• 2005

Anammox (Anaerobic Ammonium Oxidation) bacteria is recently discovered microorganism which can oxidize ammonium to nitrogen gas in the presence of nitrite under anaerobic conditions. The anammox process can save an energy for nitrification and need not require a carbon source for denitrification, however, the start-up periods takes a long time more than several months due to the long doubling time (approximately 11 days). In order to find the effects of seeding microorganisms, hydrazine, and nitrite concentration on the enhancement of the anammox activity, five kinds of microorganisms were selected. Among the several kinds of seeding microorganisms, the granule from acclimated microorganisms treating high concentration of ammonia nitrogen (A-1) and sludge from piggery wastewater treatment plant (A-2) were found to have a high anammox activity. In the case of A-1, the maximum nitrogen conversion rate represented 0.4 mg N/L-hr, and the amount of nitrite utilization was high compared to those of other seeding microorganisms. The A-4 represented a higher nitrogen conversion rate to 0.7 mg N/L-hr although the ammonium concentration in the serum bottle was high as 200 mg/L. Meanwhile, the anaerobic granule from UASB reactor treating distillery wastewater showed a low anammox activity due to the denitrification by the remained carbon sources in the granule. Hydrazine, intermediate product in anammox reaction, enhanced the anammox activity by representing 1.4 times of nitrogen gas was produced in the test bottle than that of control, when 0.4 mM of $N_2H_4$ was added to serum bottle which contains 5 mM of nitrite. The high concentration of nitrite (10 mM) resulted in the decrease of the anammox activity by showing lower production of nitrogen gas compared to that of 5 mM addition of nitrite concentration. As a result of FISH (Florescence In-Situ Hybridization) experiment, the Amx820 probe showed a more than 13% of anammox bacteria in a granule (A-1).

• 한국물환경학회지
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• 제23권3호
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• pp.397-402
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• 2007

Anammox reactor seeded with sewage sludge from RBC reactor and anaerobic granule from full-scale UASB reactor treating distillery wastewater was operated. Mixed granule and suspended sludge in the ammonium oxidizing process were taken and analyzed to investigate microbial community structure by molecular methods such as gene cloning and phylogenetic tree analysis after 250 days of continuous cultivation. The average nitrogen removal rate showed $0.9kg\;N/m^3-day$ after 250 days of continuous operation, then the maximum nitrogen removal rate showd $1.9kg\;N/m^3-day$ when $2.1kg\;N/m^3-day$ of nitrogen loading rate was applied. As results of gene cloning and phylogenetic tree analysis, Three kinds of phylum were found to be Proteobacteria, Acidobacteria and Planctomycetes (anammox bacteria) in mixed granule. Five kinds of phylum were found to be Proteobacteria, Chlorobi, Planctomycetes, Nitrospirae and Verrucomicrobia in suspended sludge. We found planctomycete KSU-1 and putative new anammox bacteria in the reactor. Microbial structure represented different consortia depending on the types of sludge in the anammox reactor.

• 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.

• Biotechnology and Bioprocess Engineering:BBE
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• 제9권5호
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• pp.345-351
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• 2004

We investigated the anaerobic ammonium oxidation (anammox) reaction in a lab-stale upflow anaerobic sludge blanket (UASB) reactor. Our aim was to detect and enrich the organisms responsible for the anammox reaction using a synthetic medium that contained low concentrations of substrates (ammonium and nitrite). The reactor was inoculated with granular sludge collected from a full-scale anaerobic digestor used for treating brewery wastewater The experiment was performed during 260 days under conditions of constant ammonium concentration ($50\;mg\;NH_4^+-N/L$) and different nitrite concentrations ($50{\~}150\;mg\;NO_2-N/L$). After 200 days, anammox activity was observed in the system. The microorganisms involved in this anammox reaction were identified as Candidatus B. Anammoxidans and K. Stuttgartiensis using fluorescence in situ hybridization (FISH ) method.

• 상하수도학회지
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• 제35권2호
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• pp.135-142
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• 2021

The activity of anaerobic ammonium oxidation (ANAMMOX) immobilized in synthetic media (Poly Ethylene Glycol, PEG) and granular form was evaluated comparatively to investigate the effect of influent nitrogen concentration and exposure of oxygen. In ANAMMOX granule reactor, when concentration of influent total nitrogen increased to 500mg/L, removal efficiency of ammonium, nitrite and nitrate were shown to 90.5±6.5, 96.6±4.9, and 93.2±6.1%, respectively. In the case of the PEG gel, it showed lower nitrogen removal performance, resulting in that the removal efficiency of ammonium, nitrite and nitrate were shown to 83.3±13.0, 96.4±6.1, and 90.3±7.5%, respectively. In second step, when exposed to oxygen, the nitrogen removal performance in the ANAMMOX granule reactor also remained stable, but the activity of PEG gel ANAMMOX was found to be inhibited. Consequently, the PEG gel ANAMMOX was a higher sensitivity than that of granular ANAMMOX with two variables applied in this study.

• Journal of Microbiology and Biotechnology
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• 제24권7호
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• pp.879-887
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• 2014

Bacteria that mediate the anaerobic oxidation of ammonium (anammox) have been detected in natural ecosystems, as well as various wastewater treatment systems. In this study, sludge from a particular landfill leachate anaerobic treatment system was selected as the incubation seed for anammox microorganism enrichment owing to its possible anammox activity. Transmission electron microscopy observation, denaturing gradient gel electrophoresis analysis, and cloning/sequencing techniques were applied to identify the diversity of anammox microorganisms throughout the incubation. During the early stage of operation, the diversity of anammox microorganisms was similar to the original complex microbes in the seed sludge. However, as incubation time increased, the anammox microorganism diversity within the system that was originally dominated by Candidatus (Ca.) Brocadia sp. was replaced by Ca. Anammoxoglobus propionicus. The domination of Ca. Anammoxoglobus propionicus produced a stable removal of ammonia (70 mg-N/l) and nitrite (90 mg-N/l), and the total nitrogen removal efficiency was maintained at nearly 95%. The fluorescence in situ hybridization results showed that Ca. Anammoxoglobus propionicus was successfully enriched from $1.8{\pm}0.6%$ initially to $65{\pm}5%$ after 481 days of operation. Therefore, the present results demonstrated the feasibility of enriching Ca. Anammoxoglobus propionicus from leachate sludge, even though the original cell count was extremely low. Application of this seldom found anammox organism could offer an alternative to current ammonia-nitrogen treatment.

• 대한환경공학회지
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• 제37권6호
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• pp.332-339
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• 2015

질소화합물은 부영양화 등 수질을 악화시키는 결과를 초래하므로 질소 제거는 수처리에 있어 가장 중요한 문제들 중 하나이다. 본 연구에서는 독립영양탈질 공정인 CANON (Completely Autotrophic Nitrogen-removal Over Nitrite)을 이용하여 암모니아성 질소 제거 효율을 평가하고, 미생물 군집 분석을 수행하였다. AOB (Ammonium Oxidizing Bacteria)와 ANAMMOX(ANaerobic AMMonium OXidation)균을 동시에 식종하고, $37^{\circ}C$에서 유입 암모니아성 질소농도 100 mg-N/L와 아질산성 질소 농도 100 mg-N/L 조건으로 운전한 결과, 성공적인 CANON 반응이 유도되었다. 유입수에서 아질산성 질소를 제외시키고 암모니아성 질소(100 mg-N/L)만을 공급하였을 때, DO농도 0.4 mg/L 이상에서는 CANON의 성능이 악화되었지만, DO농도를 0.3 mg/L으로 낮추자 71.3%의 총 질소제거효율을 나타내었다. 유입 암모니아성 질소 농도를 50 mg-N/L로 낮추었을 때, 질소 제거효율이 급격히 악화되었다. 그러나 유입농도를 다시 100 mg-N/L로 증가시키자 14일 만에 이전의 질소제거성능을 회복하였고, 이후 $76.1{\pm}4.9%$의 총 질소제거효율을 나타냈다. 온도를 상온($20{\pm}1^{\circ}C$) 조건으로 전환하자 초기에는 불안정한 CANON 반응이 일어났지만, 23일 이후에는 안정적인 총 질소제거효율($70.0{\pm}2.6$%)을 유지하였다. PCR-DGGE를 이용한 미생물군집 분석 결과, 식종원과 CANON의 미생물군집은 확연한 차이를 나타냈지만, CANON의 각 조건에 따른 미생물군집은 크게 다르지 않았다. 따라서 질소제거 성능의 악화는 미생물군집을 구성하는 미생물종의 변화에 기인하기 보다는 구성 미생물종들의 질소제거 활성의 저하에 기인하는 것으로 생각된다. 이러한 결과는 AOB와 ANAMMOX균을 식종하여 CANON 반응을 성공적으로 유도한다면, 이후 농도나 온도의 변화에도 안정적인 미생물군집을 유지할 수 있다는 것을 의미한다.