• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.5
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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.

• Journal of Korean Society of Environmental Engineers
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• v.28 no.3
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• pp.300-307
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• 2006

Anaerobic ammonium oxidation(ANAMMOX) is a novel process fur treatment of piggery waste with strong nitrogen. In this study, we investigated acid fermentation of organic matter, denitrificatiot reduction of sulfur compounds and P crystalization by hydroxyapatite during the treatment of wastewater with high strength of ammonium and organic matters by ANAMMOX process. Also, functions of hydroxylamine and hydrazine as intermedeates of ANAMMOX process were tested. This study reveals that various complex-reactions with anaerobic ammonium oxidation of piggery waste are happened and hydroxylamine and hydrazine play an important role in ANAMMOX reaction.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.286-289
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• 2001

Anaerobic ammonium oxidation with nitrite to $N_2$(anammox) is a recently discovered microbial reaction with interesting potential for nitrogen removal from wastewater. Here we investigated the microbial community structure in the sequencing batch reactor(SBR) with an anammox activity. The SBR was optimized for the enrichment of a very slowly growing microbial community and showed that possibility of anaerobic ammonium oxidation. Fluorescence in situ hybridization(FISH) analysis revealed that anaerobic ammonium oxidizers were Candidatus Brocadia anammoxidans and Candidatus Kuenenia stuttgartiensis. Furthermore, Nitrosomol1as spp. of the ${\beta}$ -subclass of Proteobacteria was also present within the anaerobic SBR microorganisms.

• Journal of Korean Society of Water and Wastewater
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• v.35 no.2
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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.

• Environmental Engineering Research
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• v.11 no.4
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• pp.173-180
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• 2006

Autotrophic nitrogen removal and its microbial community from a laboratory scale upflow anaerobic sludge bed reactor were characterized with dynamic behavior of nitrogen removal and sequencing result of molecular technique (DNA extraction, PCR and amplification of 16S rDNA), respectively. In the experiment treating inorganic wastewater, the anaerobic granular sludge from a full-scale UASB reactor treating industrial wastewater was inoculated as seed biomass. The operating results revealed that an addition of hydroxylamine would result in lithoautotrophic ammonium oxidation to nitrite/nitrate, and also hydrazine would play an important role for the success of sustainable nitrogen removal process. Total N and ammonium removal of 48% and 92% was observed, corresponding to nitrogen conversion of 0.023 g N/L-d. The reddish brown-colored granular sludge with a diameter of $1{\sim}2\;mm$ was observed at the lower part of sludge bed. The microbial characterization suggests that an anoxic ammonium oxidizer and an anoxic denitrifying autotrophic nitrifier contribute mainly to the nitrogen removal in the reactor. The results revealed the feasibility on development of high performance lithoautotrophic nitrogen removal process with its microbial granulation.

• Journal of Korean Society on Water Environment
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• v.22 no.3
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• pp.482-491
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• 2006

The anaerobic ammonium oxidation (ANAMMOX) from substrates with various $NO_2-N$ and $NH_4-N$ concentationes, which were generated from piggery waste was accomplished by using anaerobic granular sludge as seeding sludge. As the result of operation, when $NO_2-N/NH_4-N$ ratios of ANAMMOX influent were 0.6~1.5, $NO_2-N/NH_4-N$ removal ratios were exhibited 1.19~2.07 (average 1.63). The higher influent $NO_2-N/NH_4-N$ ratios resulted in higher $NO_2-N/NH_4-N$ removal ratios by ANAMMOX. It means that $NO_2-N$ concentration is very important factor in ANAMMOX. Specific ammonium removal rate was constantly as $0.03{\sim}0.04gNH_4-N/g$ VSS-day at $35^{\circ}C$ while it was $0.01gNH_4-N/g$ VSS-day at $20{\sim}30^{\circ}C$. Thus, in order to reduce the effluent N concentration, either an increase of ANAMMOX reactor HRT or more biomass accumulation at the optimal temperature can be considered.

• Journal of Korean Society of Environmental Engineers
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• v.30 no.7
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• pp.700-704
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• 2008

Nitrite and free ammonia have been known as substrate inhibitors in anaerobic ammonium oxidation. To reduce inhibitory effect of these substrates, the NH$_3$-N/NO$_2$-N ratio in the influent could be properly controlled in anaerobic ammonium oxidation process. Five kinds of NH$_3$-N/NO$_2$-N ratio were assayed in batch to find optimum NH$_3$-N/NO$_2$-N ratio, curtailing substrate inhibition. As the results of batch test, the highest T-N removal efficiency of 88% was obtained at 1.00 : 1.30 of NH$_3$-N/NO$_2$-N ratio. In addition, rate constants for ammonium and nitrite in zero-order kinetics were found to be the highest values as 7.66 mg/L$\cdot$hr and 11.89 mg/L$\cdot$hr at 1.00 : 1.30 ratio, respectively. However, as for the specific anammox activity, the ratio of NH$_3$-N/NO$_2$-N ratio was recommended as 1 : 1.15 which can maintain the highest SAA during continuous operation and preclude the accumulation of nitrite in the reactor.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.3
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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.

• Journal of Korean Society on Water Environment
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• v.36 no.3
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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.

• Journal of Korean Society on Water Environment
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• v.22 no.4
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• pp.599-604
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• 2006

Nitrogen removal with the combined SHARON (Single reactor system for high ammonium removal over nitrite)ANAMMOX (Anaerobic ammonium oxidation) process using the effluent of ADEPT (Anaerobic digestion elutriated phased treatment) slurry reactor with very low C/N ratio for piggery waste treatment was investigated. For the preceding SHARON reactor, ammonium nitrogen loading and removal rate were $0.97kg\;NH_4-N/m^3_{reactor}/day$ and $0.68kg\;NH_4-N/m^3_{reactor}/day$ respectively. In steady state, bicarbonate alkalinity consumption for ammonium nitrogen converted to $NO_2-N$ or $NO_3-N$ was 8.4 gram per gram ammonium nitrogen. The successive ANAMMOX reactor was fed with the effluent from SHARON reactor. The loading and removal rate of the soluble nitrogen defined as the sum total of $NH_4-N$, $NO_2-N$ and $NO_3-N$ in ANAMMOX reactor were $1.36kg\;soluble\;N/m^3_{reactor}/day$ and $0.7kg\;soluble\;N/m^3_{reactor}/day$, respectively. The average $NO_2-N/NH_4-N$ removal ratio by ANAMMOX was 2.41. Fluorescence in situ hybridization (FISH) analysis verified that Candidatus Kuenenia stuttgartiensis were dominate, which means that they played an important role of nitrogen removal in ANAMMOX reactor.