• 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

• 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.

• 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 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.

• 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.

• Journal of Korean Society on Water Environment
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• v.24 no.3
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• pp.383-390
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• 2008

Recently, the interests on economical nitrogen removal from wastewater are growing. As a method of the novel nitrogen removal technology, nitrogen removal via nitrite pathway by selective inhibition of free ammonia and free nitrous acid on nitrite oxidizing bacteria have been intensively studied. The inhibition effects of free ammonia and free nitrous acid are low when domestic wastewater is used, however, because of its relatively lower nitrogen concentration than the wastewater from industry and landfill, etc. In this study, a sequencing batch reactor (SBR) is proposed for nitrogen removal to investigate the effect of the low nitrogen concentration on nitrite accumulation. Nitrification efficiency reached almost 100% during the aerobic cycle and the maximum specific nitrification rate ($V_{max,nit}$) reached $17.8mg\;NH_4{^+}-N/g\;MLVSS{\bullet}h$. During the anoxic cycle, average denitrification efficiency reached 87% and the maximum specific denitrification rate ($V_{max,den}$) reached $9.8mg\;NO_3{^-}-N/g\;MLVSS{\bullet}h$. From the analysis the main reason of nitrite accumulation in the SBR was free nitrous acid rather than free ammonia. Nitrite accumulation increased with the decrease of organic content in the wastewater and the mechanism is not well understood yet. From the result of fluorescent in situ hybridization, the distribution of nitrite oxidizing bacteria was in equilibrium with ammonium oxidizing bacteria when nitrite accumulation did not occur.

• Journal of Environmental Science International
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• v.21 no.12
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• pp.1455-1462
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• 2012

The vertical distributions of nitrifying bacteria in aerobic fixed biofilm were investigated to evaluate the relationship between nitrification performance and microbial community at different HRT. Fluorescent in situ hybridization (FISH) and portable ion selective microelectrode system were adopted to analyze microbial communities and ions profiles according to the biofilm depth. Cilia media packed MLE (Modified Ludzack-Ettinger) like reactor composed of anoxic, aerobic I/II was operated with synthetic wastewater having COD 200 mg/L and $NH_4{^+}$-N mg/L at HRT of 6 hrs and 4 hrs. Total biofilm thickness of aerobic I, II reactor at 4 hrs condition was over two times than that of 6 hrs condition due to the sufficient substrate supply at 4 hrs condition (6 hrs; aerobic I 380 ${\mu}m$ and II 400 ${\mu}m$, 4 hrs; aerobic I 830 ${\mu}m$ and II 1040 ${\mu}m$). As deepen the biofilm detection point, the ratio of ammonia oxidizing bacteria (AOB) was decreased while the ratio of nitrite oxidizing bacteria (NOB) was maintained similar distribution at both HRT condition. The ratio of AOB was higher at 4 hrs than 6 hrs condition and $NH_4{^+}$-N removal efficiency was also higher at 4 hrs with 89.2% than 65.4% of 6 hrs. However, the ratio of NOB was decreased when HRT was reduced from 6 hrs to 4 hrs and $NO_2{^-}$-N accumulation was observed at 4 hrs condition. Therefore, it is considered that insufficient HRT condition could supply sufficient substrate and enrichment of AOB in all depth of fixed biofilm but cause decrease of NOB and nitrite accumulation.

• Korean Journal of Microbiology
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• v.44 no.1
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• pp.29-36
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• 2008

Nitrogen is one of the major pollutants that should be removed by wastewater treatment systems. Biological nitrogen removal (BNR) is a key technology in advanced wastewater treatment systems operated by bacterial populations. Nitrification is the first step of microbiological processes in BNR system. Ammonia is oxidized to nitrite by ammonia-oxidizing bacteria (AOB) and then nitrite is subsequently oxidized to nitrate by nitrite-oxidizing bacteria (NOB). The diversity of NOB in nitrification reactors of 3 BNR systems, Edited biological aerated filter system, Nutrient removal laboratory system, and the Rumination type sequencing batch reactor system, was investigated by terminal restriction fragment length polymorphism (T-RFLP) analysis of 16S rRNA genes. Cluster analysis of T-RF profiles showed that communities of Nitrobacter group in each system were different depending upon the process of systems. However, the clusters of Nitrospira group were divided by the habitat of aqueous and solid samples.

• Journal of Korean Society of Environmental Engineers
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• v.35 no.1
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• pp.57-63
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• 2013

A combined process consisted of a biofilm reactor and a continuously stirred-tank reactor (CSTR) was investigated for highly loaded ammonium wastewater treatment via nitrite accumulation. To enhance ammonium oxidizing bacteria over nitrite oxidizing bacteria on the surface of carriers, the biofilm reactor was operated at temperature of $35^{\circ}C$ for more than three months but the influent ammonium (500 mg-N/L) was partially oxidized to nitrite (240 mg-N/L). As pH was increased from 7.5 to 8.0, nitrite accumulation was fully achieved due to the inhibition of nitrite oxidizing bacteria under high free ammonia concentration. The biofilm reactor performance was severely deteriorated at the hydraulic retention time of 12 hr, at which incomplete nitrification of ammonia was observed. Various solubilization methods were applied to sewage sludge for enhancing its biodegradability and the combined method, alkaline followed by ultrasonic, gave the highest solubilization efficiency (58%); the solubilized solution was used as the external carbon source for denitrification reaction in CSTR. FISH analysis showed that the dominant microorganisms on the carriers were ammonium oxidizing bacteria such as Nitrosomonas spp. and Nitrospirar spp. but low amounts of nitrite oxidizing bacteria as Nitrobacter spp. was also detected.

• Journal of the Korea Organic Resources Recycling Association
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• v.18 no.1
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• pp.73-80
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• 2010

Two nitrite oxidizing bacteria, NOB1 and NOB2, were isolated from anaerobic digester liquer of food wastewater and analyzed for their growth characteristics and the ability to oxidize nitrite under different temperature, pH, and DO( dissolved oxygen) concentrations. Both of the isolated strains have shown the best growth at pH 7.0 and at $35^{\circ}C$, and also shown higher growth rate with the increasing dissolved oxygen concentrations. As the factors to restrict the growth of these strains, parameters such as pH and DO were found to be effective ones, by increasing (up to 9.0) or decreasing pH (up to 5.0), or lowing DO below 1.0 ppm. Especially, the ability to oxidize nitrite in both strains was about 50% lower in below 1.0 ppm of DO than above of 1.0 ppm. NOB2 was found to be two times greater in both the growth rate and the nitrite-oxidizing ability than NOB1.