• Journal of Korean Society on Water Environment
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• v.31 no.4
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• pp.399-406
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• 2015

A sequencing batch reactor was operated under different pH conditions to see the influence of free ammonia (FA) and free nitrous acid (FNA) to microbial community on ammonium partial nitrification. Long-term influences of FA and FNA were evaluated by polymerase chain reaction-denaturing gradient gel electrophoresis and fluorescence in situ hybridization. Nitrite accumulation was successfully achieved at pH 8.2 and 6.3. The shifts in the microbial community were observed when influent ammonia concentration increased to 1 g $NH_4$-N/L at pH 8.2, and then when pH was dropped to 6.3. Both Nitrosomonas and Nitrosospira were selected during the startup of the reactor, and eventually became dominant members as ammonia-oxidizing bacteria. The results of molecular microbiological analysis strongly suggested that the composition of microbial community was changed according to the method used to control nitrite-oxidizing bacteria.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.5
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• pp.323-326
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• 2002

To detect whole ammonia-oxidizing bacteria in the activated sludge, group-specific primers targeting the 16S-rRNA gene of ammonia-oxidizing bacteria were used. The electrophoresis pattern of the PCR products seemed to produce a single band of approximately 1.0 k bp for the bacteria in activated sludge and Nitrosomonas europaea. No band was observed for nitrite-oxidizer Nitrobacter winogradskyi and heterotrophs such as Pseudomonas putida. Then direct measurement of the PCR product was made by fluorometry using the reagent Hoechist 33258, so that the fluorescent intensity was in proportional to the cell number of the sample up to 240. Total time required for the test was about 4 h including DNA extraction. The DNA fragments produced were cloned and their sequences showed high similarity to those of Nitrosomonas spp. This study showed the feasibility to detect ammonia-oxidizing bacteria and to esti-mate their population rapidly for the control of the nitrogen elimination process.

• Journal of Soil and Groundwater Environment
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• v.21 no.4
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• pp.50-59
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• 2016

Spatial and temporal changes in nitrification activities and distribution of microbial population of ammonia oxidizing bacteria (AOB) and ammonia oxidizing archaea (AOA) in paddy soils were investigated. Soil samples were collected in March and October 2015 from rice paddy with and without the presence of confined animal feeding operations. Incubation experiments and quantitative polymerase chain reaction showed that AOA's contribution to nitrification kinetics was much higher in locations where organic nitrogen in animal waste is expected to significantly contribute to overall nitrogen budget, and temporal variations in nitrification kinetics were much smaller for AOA than AOB. These differences were interpreted to indicate that different microbial responses of two microbial populations to the types and concentrations of nitrogen substrates were the main determining factors of nitrification processes in the paddy soils. The copy numbers of ammonium monooxygenase gene showed that AOA colonized the paddy soils in higher numbers than AOB with stable distribution while AOB showed variation especially in March. Although small in numbers, AOB population turned out to exert more influence on nitrification potential than AOA, which was attributed to higher fluctuation in AOB cell numbers and nitrification reaction rate per cells.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.282-285
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• 2001

The microbial community structure and in situ spatial distribution of ammonia oxidizing and nitrite oxidizing bacteria in nitrifying biofilm of an upflow biological aerated filter system were investigated. The reactor had been continuously operated under high free ammonia concentration and low DO concentration for nitrite accumulation more than 2 years before the experiment. Fluorescence in situ hybridization

• Environmental Engineering Research
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• v.21 no.2
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• pp.140-144
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• 2016

Owing to the kinetic differences in ammonia oxidation among ammonia-oxidizing microorganisms (AOM), there is no standard set of kinetic values that can be used as a representative set for nitrifying wastewater treatment plant (WWTP) design. As a result, this study clarified a link between the half-saturation constants for ammonia oxidation (Ks) and the dominant ammonia-oxidizing bacterial (AOB) groups in sludge from full-scale WWTPs and laboratory-scale nitrifying reactors. Quantitative polymerase chain reaction analyses revealed that AOB affiliated with the Nitrosomonas oligotropha cluster were the dominant AOM groups in the sludge taken from the low-ammonia-level WWTPs, while AOB associate with the Nitrosomonas europaea cluster comprised the majority of AOM groups in the sludge taken from the high-ammonia-level WWTPs and nitrifying reactors. A respirometric assay demonstrated that the ammonia Ks values for the high-ammonia-level WWTPs and nitrifying reactors were higher than those of the low-ammonia-level plants. Using the Ks values of available AOM cultures as a reference, the Ks values of the analyzed sludge were mainly influenced by the dominant AOB species. These findings implied that.different sets of kinetic values may be required for WWTPs with different dominant AOM species for more accurate WWTP design and operations.

• Journal of the Korea Organic Resources Recycling Association
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• v.8 no.2
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• pp.146-153
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• 2000

To elucidate succession of bacterial populations, especially nitrifying bacteria during the composting of cattle manure with apple pomace, fluorescent in situ hybridization(FISH) using rRNA targeted oligonucleotide probes were applied. The density of ammonia-oxidizing bacteria was ranged from $3,3{\times}10^6cells/g$ dw to $13,4{\times}10^6cells/g$ dw with the peak value after 26 composting days whereas that of nitrite-oxidizing bacteria varied between $6.0{\times}10^6cells/g$ dw and $17.2{\times}10^6cells/g$ dw with the peak value after 7 composting days. And the tendency that the numbers of nitrite-oxidizing bacteria were higher than those of ammonia-oxidizing bacteria, and the peak-time of their densities were the same as that of data determined by the ratio of ammonia-oxidizing bacteria and nitrite-oxidizing bacteria to eubacteria. The peak of ammonia-oxidizing bacteria followed the peak of nitrite-oxidizing bacteria, at the late phase of composting process could be probably caused by the depletion of volatile ammonia of composting materials. Besides these results indicate that FISH method is a useful tool for detection of slow growing nitrifying bacteria.

• 제어로봇시스템학회:학술대회논문집
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• 2000.10a
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• pp.287-287
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• 2000

As a method to control the bacteria number in adequate level, a real time control system based on microscope image processing measurement for the bacteria is adopted. For the experiment, Ammonia-oxidizing bacteria such as Acinetobacter sp. are used. This paper proposed hybrid method combined watershed algorithm with adaptive automatic thresholding method to enhance segmentation efficiency of overlapped image. Experiments was done to show the effectiveness of the proposed method compared to traditional Otsu's method, Otsu's method with adaptive automatic thresholding method and human visual method.

• Journal of Korean Society on Water Environment
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• v.24 no.2
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• pp.214-220
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• 2008

Nitrifying bacteria were detected by fluorescent in situ hybridization (FISH) method at 6 sampling sites with different eutrophication degree in the Nak-Dong River and their tributaries. And conventional physico-chemical parameters including $NH_4-N$, $NO_3-N$, and TN were determined concurrently. In rainy period (July), there was no noticeable difference between the number of ammonia/nitrite-oxidizing bacteria detected at each site except Sang-Ju and the ratio of nitrifying bacteria to total counts stained by DAPI varied in 6~33%. By contrast, in the dry period (October), both of bacterial population was increased differently and the ratio of nitrifying bacteria to total counts ranged more widely from 6% in heavily polluted water zone, Hwa-Won to 60% in upper tributary with high agricultural land use. Byung-Sung-Chun. In January, the numbers of ammonia-oxidizing bacteria was reduced up to one tenth, while those of nitrite-oxidizing bacteria was apparently increased maybe due to high DO and low DOC.

• Journal of Korean Society of Water and Wastewater
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• v.26 no.3
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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.

• Environmental Engineering Research
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• v.13 no.3
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• pp.125-130
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• 2008

Free ammonia ($NH_3$-N) inhibition of nitrite-oxidizing bacteria (NOB) has been widely studied for partial nitrification (or nitrite accumulation) and denitrification via nitrite ($NO_2^-$-N) as a low-cost treatment of ammonium containing wastewater. The literature on $NH_3$-N inhibition of NOB, however, shows disagreement about the threshold $NH_3$-N concentration and its degree of inhibition. In order to clarify the confusion, a simple and cheap respirometric method was devised to investigate the effect of free ammonia inhibition of NOB. Sludge samples from an autotrophic nitrifying reactor were exposed to various $NH_3$-N concentrations to measure the maximum specific nitrite oxidation rate ($\hat{K}_{NO}$) using a respirometer. NOB biomass was estimated from the yield values in the literature. Free ammonia inhibition of nitrite oxidizing bacteria was reversible and the specific nitrite oxidation rate ($K_{NO}$) decreased from 0.141 to 0.116, 0.100, 0.097 and 0.081 mg $NO_2^-$-N/mg NOB h, respectively, as the $NH_3$-N concentration increased from 0.0 to 1.0, 4.1, 9.7 and 22.9 mg/L. A nonlinear regression based on the noncompetitive inhibition mode gave an estimate of the Inhibition concentration ($K_I$) of free ammonia to be 21.3 mg $NH_3$-N/L. Previous studies gave $\hat{K}_{NO}$ of Nitrobacter and Nitrospira as 0.120 and 0.032 mg/mg VSS h. The free ammonia concentration which inhibits Nitrobacter was $30{\sim}50\;mg$ $NH_3$-N/L and Nitrospira was inhibited at $0.04{\sim}0.08\;mg$ $NH_3$-N/L. The results support the fact that Nitrobacter is the dominant NOB in the reactor. The variations in the reported values of free ammonia inhibition may be due to the different species of nitrite oxidizers present in the reactors. The respirometric method provides rapid and reliable analysis of the behavior and community of the nitrite oxidizing bacteria.