• Korean Journal of Environmental Agriculture
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• v.39 no.1
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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.

• Journal of Korean Society of Environmental Engineers
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• v.28 no.10
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• pp.1055-1064
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• 2006

Extremely slow growing anammox(anaerobic ammonium oxidation) bacteria were cultivated using a combination of UASB(Upflow Anaerobic Sludge Blanket) reactor seeded with anaerobic granular sludge and carbon-fiber cultivating reactor. After 180 days of continuous cultivation, average nitrogen removal rate showed 0.54 kg $N/m^3-day$ when 0.6 kg $N/m^3-day$ of nitrogen loading was applied. The black granule was changed to brown and red granule as continuous operation, and the red granule was highly dependant on the high anammox activity. Microbial community structure of red granule in the UASB reactor was analyzed by molecular methods such as gene cloning, phylogenetic tree analysis, and FISH(Fluorescence In Situ Hybridization) method. As a result of gene cloning and phylogenetic tree analysis, 5 kinds of phylum were found to be Planctomycetes, Proteobacteria, Acidobacteria, Chlorobi and Chloroflexi. 13 clones were matched to anammox bacteria among 51 clones in the red anammox granule. In-silico test which used cloning information and FISH probe of the AMX368 was conducted to detect the presence of anammox bacteria in the red anammox granule. As a result of in-silico test only one clone was exactly matched to AMX368 but 11 clones was mutated one base among 18 bases representing all 12 clones are anammox bacteria. A filamentous Chloroflexi might be related to the granulation of anammox bacteria. As a result of FISH analysis, anammox bacteria was abundant in the red anammox granule.

• Journal of Korean Society of Water and Wastewater
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• v.28 no.2
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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 Korean Society on Water Environment
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• v.34 no.1
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• pp.96-108
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• 2018

Large efforts have recently been made on research and development of sustainable and energy-efficient short-cut nitrogen removal processes owing to strong attention to the energy neutral/positive wastewater treatment system. Anaerobic ammonium oxidizing bacteria (anammox bacteria) have been highlighted since 1990's due to their unique advantages including 60% less energy consumption, nearly 100% reduction for carbon source requirement, and 80% less sludge production. Side-stream short-cut nitrogen removal using anammox bacteria and partial nitritation anammox (PN/A) has been well established, whereas substantial challenges remain to be addressed mainly due to undesired main-stream conditions for anammox bacteria. These include low temperature, low concentrations of ammonia, nitrite, free ammonia, free nitrous acid or a combination of those. In addition, an anammox side-stream nitrogen management is insufficient to reduce overall energy consumption for energy-neutral or energy positive water resource recovery facility (WRRF) and at the same time to comply with nitrogen discharge regulation. This implies the development of the successful main-stream anammox based technology will accelerate a conversion of current wastewater treatment plants to sustainable water and energy recovery facility. This study discusses the status of the research, key mechanisms & interactions of the protagonists in the main-stream PN/A, and control parameters and major challenges in process development.

• Journal of Korean Society of Water and Wastewater
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• v.32 no.5
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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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• v.27 no.7
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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.

• Journal of Korean Society of Environmental Engineers
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• v.31 no.10
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• pp.919-926
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• 2009

Enrichment of anaerobic ammonium oxidation (ANAMMOX) bacteria is the essential step for operating full-scale ANAMMOX bioreactor because adding a significant amount of seeding sludge is required to stabilize the ANAMMOX reactor. In this study, the enrichment of ANAMMOX bacteria from an activated sludge using sequencing batch reactor was conducted and verified by analyzing changes in the microbial community structure. ANAMMOX bacteria were successfully enriched for 70 days and the substrate removal efficiencies showed 98.5% and 90.7% for $NH_4\;^+$ and $NO_2\;^-$ in the activity test, respectively. The phylogenetic trees of Planctomycetes phylum showed that the diverse microbial community structure of an activated sludge was remarkably simplified after the enrichment. All 36 clones, obtained after the enrichment, were affiliated with ANAMMOX bacteria of Candidatus Brocadia (36%) and Candidatus Anammoxoglobus (64%) genera. The quantification using real-time quantitative PCR (RTQ-PCR) revea ed that the 16S rDNA concentration of ANAMMOX bacteria was 74.8% compared to the granular ANAMMOX sludge obtained from an upflow ANAMMOX sludge bed reactor which had been operated for more than one year. The results of molecular analysis supported that the enriched sludge could be used as a seeding sludge for a full-scale ANAMMOX bioreactor.

• Journal of Korean Society on Water Environment
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• v.22 no.2
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• pp.258-263
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• 2006

ANAMMOX (Anaerobic ammonium oxidation) reactor, which was cultivated ANAMMOX bacteria in mesophilic condition ($35^{\circ}C$), was operated to investigate the effects of temperature. In $20{\sim}30^{\circ}C$ of operation condition, which was assumed as field-temperature, total N removal and $NH_4-N$ removal rate were declined from about 2.50 and $1.27kg\;N/{m^3}_{reactor}-day$ (0.06 and 0.03 kg N/kgVSS/day) to 1.62 and $0.41kg\;N/{m^3}_{reactor}-day$ (0.04 and 0.01 kg N/kgVSS/day), In this range of temperature, ANAMMOX had very low activities but acid fermentation bacteria and denitrifiers, which were competitors of substrates, had high activities relatively. Though operation temperature was higher than inhibition condition for two months, ANAMMOX activities could not been recovered once they were inhibited by low temperature. This fact was resulted from very slow doubling time of ANAMMOX bacteria. This study shows that maintenance device of optimal temperature is necessary required in field application of ANAMMOX.

• Journal of Korean Society on Water Environment
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• v.38 no.4
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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.

• Membrane and Water Treatment
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• v.10 no.4
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• pp.265-275
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• 2019

This study aimed to develop a compact partial nitritation step by forming granules with high Ammonia-Oxidizing Bacteria (AOB) fraction using the Air-lift Granulation Reactor (AGR) and to evaluate the feasibility of treating reject water with high ammonium content by combination with the Anammox process. The partial nitritation using AGR was achieved at high nitrogen loading rate ($2.25{\pm}0.05kg\;N\;m-3\;d^{-1}$). The important factors for successful partial nitritation at high nitrogen loading rate were relatively high pH (7.5~8), resulting in high free ammonia concentration ($1{\sim}10mg\;FA\;L^{-1}$) and highly enriched AOB granules accounting for 25% of the total bacteria population in the reactor. After the establishment of stable partial nitritation, an effluent $NO_2{^-}-N/NH_4{^+}-N$ ratio of $1.2{\pm}0.05$ was achieved, which was then fed into the Anammox reactor. A high nitrogen removal rate of $2.0k\; N\;m^{-3}\;d^{-1}$ was successfully achieved in the Anammox reactor. By controlling the nitrogen loading rate at the partial nitritation using AGR, the influent concentration ratio ($NO_2{^-}-N/NH_4{^+}-N=1.2{\pm}0.05$) required for the Anammox was controlled, thereby minimizing the inhibition effect of residual nitrite.