• 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 Korean Society of Water and Wastewater
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• v.26 no.4
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• pp.591-598
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• 2012

Denitrification is an important biological mechanism in wastewater treatment process because this process is technically to remove nitrogen from water to air. There have been lots of study about denitrification engineering and molecular biological research about denitrifying bacteria, respectively. However, combination of these researches was unusual and rare. This study is about the correlation between quantity of denitrifying bacteria and denitrification potential, and consists of NUR batch test as analysis method of denitrification potential and quantitative molecular analysis for denitrifying bacteria. Three reactors (A/O, MLE and A/O of nitrogen deficiency) are operated to get activated sludge with various denitrification potential. All samples which were acquired from reactors were measured denitrification potential by NUR test and NUiR test. Also, Real-time PCR was conducted for quantification of denitrifying bacteria composition in activated sludge. The various denitrification potentials were measured in the reactors. The denitrifiaction potential was the highest in MLE process and the reactor of the nitrogen deficiency showed the lowest. Genomic DNA of activated sludge was obtained and consequently, real-time PCRuse the primer sets of nirK and nirS　were conducted to quantify genes involving denitrification reductase production. As the result of real-time PCR, nirK gene showed more significant influence on denitrification potential comapred with nirS gene.

• Korean Journal of Microbiology
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• v.53 no.1
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• pp.9-19
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• 2017

We studied soil composition, $N_2O$ production, a number of denitrifying bacteria, community structure and T-RFLP patterns of denitrifying bacteria dependent on agricultural methods with the change of seasons. Analyses of the soil chemical composition revealed that total carbon and total organic carbon contents were 1.57% and 1.28% in the organic farming soil, 1.52% and 1.24% in the emptiness farming soil, and 1.40% and 0.95% in traditional farming soil, respectively. So, the amount of organic carbon was relatively high in the environment friendly farming soils than traditional farming soils. In case of $N_2O$ production, the amount of $N_2O$ production was high in May and November soils, but the rate of $N_2O$ production was fast in August soil. The average number of denitrifying bacteria were $1.32{\times}10^4MPN{\cdot}g^{-1}$ in the organic farming soil, $1.17{\times}10^4MPN{\cdot}g^{-1}$ in the emptiness farming soil, and $6.29{\times}10^3MPN{\cdot}g^{-1}$ in the traditional farming soil. It was confirmed that the environment friendly farming soil have a larger number of denitrifying bacteria than the traditional farming soil. As a result of the phylogenetic analyses, it was confirmed that six clusters were included in organic farming soil among total 10 clusters. And the result of PCA profile distribution of T-RFLP pattern on agricultural methods, the range of distribution showed wide in the organic farming method, relatively narrow in the conventional farming method, and middle in the emptiness farming method. Therefore, we could concluded that the distribution and the community structure of denitrifying bacteria were changed according to the agricultural methods and seasons.

• Korean Journal of Microbiology
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• v.38 no.2
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• pp.113-118
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• 2002

Laboratory experiments were aimed to evaluate the effect of nitrate as a electron acceptor during the biological phosphorus uptake and to investigate the microbial community. Anaerobic-anoxic sequencing batch reactor (SBR) compared the removal behaviour to anaerobic-oxic SBR, both SBRs maintained lower effluent quality with 1.0 mgp/1. Anaerobic-anoxic SBR was able to remove additional 5.0 to 7.0 mg (P+N)/ι than other biological nutrient removal (BM) system. Therefore, it was proposed that the anaerobic-anoxic SBR was more effective at weak sewage. From the results of the maicrobial community analysis, it can be inferred that denitrifying bacteria and polyphosphate accumulating bacteria coexist in anaerobic-anoxic SBR during stable condition for removing the nitrogen and phosphorus. Particularly, it was suggested that the Zoogloea ramigera in the $\beta$-subclass of proteobacteria and the Alcaligenes defragrans of the Rhodocyclus group in the $\beta$-subclass of proteobacteria played a major role for removing the nitrogen and phosphorus as dPAOs (denitrifying phosphate accumulating organisms).

• Journal of Korean Society on Water Environment
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• v.29 no.6
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• pp.777-781
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• 2013

Ion-exchange technology is one of the best for removing nitrate from drinking water. However, problems related to the disposal of spent brine from regeneration of exhausted resins must be overcome so that ion exchange can be applied more widely and economically, especially in small communities. In this background, a combined bio-regeneration and ion-exchange system was operated in order to prove that nitrate-laden resins could be bio-regenerated through direct contact with denitrifying bacteria. A nitrate-selective A520E resin was successfully regenerated by denitrifying bacteria. The bio-regeneration efficiency of nitrate-laden resins increased with the amount of flow passed through the ion-exchange column. When the fully exhausted resin was bio-regenerated for 5 days at the flowrate of 30 BV/hr and MLSS concentration of $125{\pm}25mg/L$, 97.5% of ion-exchange capacity was recovered. Measurement of nitrate concentrations in the column effluents also revealed that less than 5% of nitrate was eluted from the resin during 5 days of bio-regeneration. This result indicates that the main mechanism of bio-regeneration is the direct reduction of nitrate by denitrifying bacteria on the resin.

• Microbiology and Biotechnology Letters
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• v.39 no.3
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• pp.210-217
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• 2011

In an effort to isolate novel bacteria for the bioremediation of over-fertilized soils, we identified 135 denitrifying cells out of 3,471 oligotrophic bacteria pools (3.9%) using a denitrification medium supplemented with potassium nitrate as the sole nitrogen source. Soil samples were taken from ecologically well-conserved areas, including a mountain swamp around the demilitarized zone (Yongneup), two ecoparks (Upo and the Mujechi bog), and ten representative islands around the Korean peninsula (Jejudo, Daecheongdo, Socheongdo, Baekryeongdo, Ulrungdo, Dokdo, Geomundo, Hongdo, Huksando and Yeonpyeongdo). All of the 135 bacteria produced nitrogen gas from the denitrification medium, and were proved to be nitrate reductase positive by API-BioLog tests. Phylogenetic analysis using 16S rDNA sequences revealed that the 135 bacteria consisted of 44 different genera. Along with the most prominent, Proteobacteria (87.4%), we identified denitrifying bacteria from Firmicutes (9.4%), Actinobacteria (2.4%), and Bacteroidetes (0.8%). Physiological analyses of the 44 representative denitrifying bacteria, under various pH levels, growth temperatures and salt stresses, revealed 12 favorable denitrifying strains for soil bioremediation.

• Journal of Microbiology and Biotechnology
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• v.15 no.1
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• pp.14-21
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• 2005

By using PCR with nirS gene primers, three nirSharboring denitrifying bacteria (strain N6, strain N23, and strain R13) were newly isolated from activated sludge of a weak municipal wastewater treatment plant. Small-subunit rRNA gene-based analysis indicated that strain N6, strain N23, and strain R13 were closely related to Arthrobacter sp.,Staphylococcus sp., and Bacillus sp., respectively. In an attempt to identify their roles in biological nitrate and nitrite removal from sewage, we investigated their specific denitrification rates (SDNRs) for $NO_-^3$ - and $NO_-^2$ - in various cultures. All purecultures of each isolated nirS-harboring bacterial strain could remove $NO_-^3$ - and $NO_-^2$ - simultaneously in high efficiency, and the carbon requirements for $NO_-^3$ - removal of strain N6 and strain R13 were effectively low at 3.1 and 4.1 g COD/g $NO_3N$, respectively. In the case of mix-cultures of the strains (N6+N23, N6+R13, N23+R13, and N6+N23+R13), their SDNRs for $NO_-^3$ - were also effective, and their carbon requirements for $NO_-^3$ - removal were also effective at 3.0- 3.8 g COD/g NO3N. However, all tested mix-cultures accumulated $NO_-^2$ - in their culture media. On the other hand, the continuous culture of activated sludge mixed with strain N6 showed no significant increase of $NO_-^3$ - removal in comparison with strain N6's pure culture. These results suggest that nitrate and nitrite removal in biological wastewater treatment might be dependent on complicated bacterial interactions, including several effective denitrifying bacteria isolated in this study, rather than the specific bacterial types present and the number of bacterial types in activated sludge.

• Journal of Microbiology
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• v.43 no.5
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• pp.383-390
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• 2005

The diversity of the denitrifying bacterial populations in Daejeon Sewage Treatment Plant was examined using a culture-dependent approach. Of the three hundred and seventy six bacterial colonies selected randomly from agar plates, thirty-nine strains that showed denitrifying activity were selected and subjected to further analysis. According to the morphological and biochemical properties, the thirty nine isolates were divided into seven groups. This grouping was supported by an unweighted pair group method, using an arithmetic mean (UPGMA) analysis with fatty acid profiles. Restriction pattern analysis of 16S rDNA with four endonucleases (AluI, BstUI, MspI and RsaI) again revealed seven distinct groups, consistent with those defined from the morphological and biochemical properties and fatty acid profiles. Through the phylogenetic analysis using the 16S rDNA partial sequences, the main denitrifying microbial populations were found to be members of the phylum, Proteobacteria; in particular, classes Gammaproteobacteria (Aeromonas, Klebsiella and Enterobacter) and Betaproteobacteria (Acidovorax, Burkholderia and Comamonas), with Firmicutes, represented by Bacillus, also comprised a major group.

• Microbiology and Biotechnology Letters
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• v.49 no.3
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• pp.432-439
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• 2021

Microbial community plays important roles in the culture environment of mud crab Scylla serrata. One of the environmental management efforts for the cultivation of S.serrata is by stabilizing microorganisms involved in nitrogen cycle process. The availability of dissolved inorganic nitrogen in its culture environment under a recirculating system closely relates to the nitrogen cycle, which involves both anaerobic and aerobic bacterial activities. Anaerobically, there are two major nitrogen compound degradation processes, i.e., denitrification and dissimilatory nitrate reduction to ammonium (DNRA). This study aimed to identify denitrifying and DNRA bacteria isolated from the recirculating cultivation of S. serrata. The water samples were collected from anaerobic filters called close filter system, which is anaerobically conditioned with the addition of varying physical filter materials in the recirculating mud crab cultures. The results showed that three denitrifying bacterial isolates and seven DNRA bacterial isolates were successfully identified. The phylogenetic analysis based on 16S rRNA gene of the denitrifying bacteria revealed that HIB_7a had the closest similarity to Stenotrophomonas daejeonensis strain MJ03. Meanwhile, DNRA bacterial isolate of HIB_92 showed a 100% similarity to Bacillus sonorensis strain N3, Bacillus vallismortis strain VITS-17, Bacillus tequlensis strain TY5, Geobacillus sp. strain DB24, Bacillus subtilis strain A1, and Bacillus mojavensis strain SSRAI21. This study provides basic information denitrifying and DNRA bacterial isolates identity which might have the potential to be applied as probiotics in aquaculture systems in order to maintain optimal environmental conditions.

• Journal of Korean Society of Environmental Engineers
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• v.22 no.10
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• pp.1757-1763
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• 2000

To consider the nutrient removal characteristics of BNR process activating soil microorganisms under the influence of DPB and to clear the characteristics of DPB under anoxic condition was investigated in the this study. The batch tests were conducted using sludge sampled from the BNR process activating soil microorganisms during operation periods. The results of this study were summarized as follows: - The DPB(Denitrifying Phosphorus removing Bacteria) performing denitrification and phosphorus uptake in the anoxic phase plays an important role in removing nitrogen and phosphorus in the BNR process activating soil microorganisms. - The PUR(Phosphorus Uptake Rate) of DPB in the anoxic phase was to be about 50% of PUR in the aerobic phase. - The DPB in the BNR process turned out to be increasing nutrient removal efficiency of BNR process.