• Journal of Microbiology and Biotechnology
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• v.25 no.10
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• pp.1670-1679
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• 2015

Two hollow fiber membrane biofilm reactors (HF-MBfRs) were operated for autotrophic nitrification and hydrogenotrophic denitrification for over 300 days. Oxygen and hydrogen were supplied through the hollow fiber membrane for nitrification and denitrification, respectively. During the period, the nitrogen was removed with the efficiency of 82-97% for ammonium and 87-97% for nitrate and with the nitrogen removal load of 0.09-0.26 kg NH₄⁺-N/m³/d and 0.10-0.21 kg NO₃^--N/m³/d, depending on hydraulic retention time variation by the two HF-MBfRs for autotrophic nitrification and hydrogenotrophic denitrification, respectively. Biofilms were collected from diverse topological positions in the reactors, each at different nitrogen loading rates, and the microbial communities were analyzed with partial 16S rRNA gene sequences in denaturing gradient gel electrophoresis (DGGE). Detected DGGE band sequences in the reactors were correlated with nitrification or denitrification. The profile of the DGGE bands depended on the NH₄⁺ or NO₃^- loading rate, but it was hard to find a major strain affecting the nitrogen removal efficiency. Nitrospira-related phylum was detected in all biofilm samples from the nitrification reactors. Paracoccus sp. and Aquaspirillum sp., which are an autohydrogenotrophic bacterium and an oligotrophic denitrifier, respectively, were observed in the denitrification reactors. The distribution of microbial communities was relatively stable at different nitrogen loading rates, and DGGE analysis based on 16S rRNA (341f /534r) could successfully detect nitrate-oxidizing and hydrogen-oxidizing bacteria but not ammonium-oxidizing bacteria in the HF-MBfRs.

• Journal of Korean Society of Water and Wastewater
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• v.10 no.3
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• pp.55-63
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• 1996

Nitrification reaction consists of two reactions: nitritification which oxidizes ammonia nitrogen to nitrite nitrogen and nitratification which oxidizes nitrite nitrogen to nitrate nitrogen. Each reaction is carried out by Nitrosomonas and Nitrobacter, respectively. The effective maximum growth rates for both bacteria have to be determined to design aeration tank whenever the aeration tanks have to nitrify ammonia nitrogen in influent. And these values are very important to use mathematical models such as IAWPRC model to simulate nitrification in activated sludge. There are several methods to determine these valves, however, the Fed-Batch experiments can determine these values within 72 hours. In this study, the mathematical equations and experimental procedures for Fed-Batch test are presented. Also, the experimental data and reported values are compared. The estimated mean values of maximum specific growth rates for Nitrosomonas and Nitrobacter are $0.5010day^{-1}$ and $0.6704day^{-1}$, respectively.

• 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.

• 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 Environmental Science International
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• v.14 no.9
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• pp.811-815
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• 2005

The activated sludge from the aeration basin of the Su-yeong municipal wastewater treatment plant which has operated by a standard activated sludge process in Busan, Korea was investigated during April 2004 and January 2005 with several bio-indicators. The number of bacteria and fungi per gram of dry weight of MLSS were estimated to be $3.1\times10^6\sim1.5\times10^8\;and\;l.1\times10^3\sim1.1\times10^5$ colony forming units, respectively, by the plate agar method. By cultivation-independent methods, such as 4',6-diamidino-2-phenylindole stain and fluorescence in situ hybridization, the ratio of eubacteria to the entire biomass was evaluated by more than $80\%$ (v/v). The ratio of ammonia-oxidizing bacteria and nitrite-oxidizing bacteria to the total eubacteria was detennined to be $7.0\sim9.8\%\;and\;3.3\sim6.2\%$ without heavy variation in spite of a period of relatively low temperature in the basin. It would be expected that the nitrification would occur or at least co-exist throughout the year in the sludge of many municipal WWTP with influents that contain the sufficient nitrogen sources although the WWTP does not have any specialized processes for the removal of nitrogen.

• Environmental Engineering Research
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• v.24 no.3
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• pp.376-381
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• 2019

In this study, a poly(vinyl) alcohol/sodium alginate (PVA/SA) mixture was used to fabricate core-shell structured gel beads for autotrophic single-stage nitrogen removal (ASNR) using aerobic and anaerobic ammonia-oxidizing bacteria (AAOB and AnAOB, respectively). For stable ASNR process, the mechanical strength and oxygen penetration depth of the shell layer entrapping the AAOB are critical properties. The shell layer was constructed by an interfacial gelling reaction yielding thickness in the range of 2.01-3.63 mm, and a high PVA concentration of 12.5% resulted in the best mechanical strength of the shell layer. It was found that oxygen penetrated the shell layer at different depths depending on the PVA concentration, oxygen concentration in the bulk phase, and free ammonia concentration. The oxygen penetration depth was around $1,000{\mu}m$ when 8.0 mg/L dissolved oxygen was supplied from the bulk phase. This study reveals that the shell layer effectively protects the AnAOB from oxygen inhibition under the aerobic conditions because of the respiratory activity of the AAOB.

• Journal of Korean Society on Water Environment
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• v.22 no.6
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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.

• 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.

• Korean Journal of Microbiology
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• v.42 no.1
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• pp.26-33
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• 2006

To understand the efficient process of biological nitrogen removal (BNR) system, the structure of bacterial communities in nitrification reactors was analyzed using PCR and terminal restriction fragment length poly morphism (I-RFLP) methods. In this study, we used an advanced treatment system with plotting media, Nutrient Removal Laboratory system, or the rumination type sequencing batch reactor (SBR) system. The terminal restriction fragments of ammonia-oxidizing bacteria (AOB) and other $\beta-proteobacteria$ were observed in all of three BNR systems. The nucleotide sequence analysis of terminal restriction fragments showed that Nitrosomonas and Nitrosolobus were major populations of AOB in SBR system, whereas uncultured $\beta-proteobacteria$ and Cardococcus australiensis were the predominant groups in other two BNR systems. Also the SBR system may be more efficient to enrich AOB. These results indicate that the different structure of bacterial community may be developed depending on the wastewater treatment systems, although the same influent is used.

• Korean Journal of Soil Science and Fertilizer
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• v.28 no.4
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• pp.356-362
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• 1995

This experiment was carried out to investigate the effects on the seasonal population change of microflora of long-term application of organic matters in Fluvio-Alluvial plain of Jeonbug series. As organic matters, rice straw and compost of 5 and 10ton/ha, which were applied with the different nitrogen fertilizer level of 0, 150kg/ha into the soil 15cm deep, respectively. A number of total aerobic bacteria were gradually increased from just after water-logging before rice transplanting to pancle formations stage, afterthat decreased at harvest. The other side, a number of actinomycetes, fungi and cellulose-decomposers were slightly fluctuated until panicle formation stage and increased at havesting stage. In general, microorganism numbers were higher in organic matter with long-term nitrogen fertilizer applied plot, while cellulose-decomposers were higher in only organic matter applied plot. The microorganisms of ammonia-oxidizing, nitrate-reducing and nitrite-oxidizing, and denitrifying bacteria showed the maximum number at harvest stage, at panicle formation stage and at early tillering stage, respectively, while that of ammonifying bacteria were variable if nitrogen fertilizer applied or not at the respective periods in nitrogen cycle under water-logging. These bacteria were numerous in the organic matter plots combined with nitrogen fertilizer, especially, denitrifying bacteria in rice straw, others no difference.