• Environmental Engineering Research
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• v.20 no.1
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• pp.51-57
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• 2015

As expected, the expression of denitrifying genes in a Typha wetland (relatively stagnant compared to other ponds), showing higher nitrogen removal efficiency in summer, was affected by temperature. The abundance and gene transcripts of nitrate reductase (narG), nitrite reductase (nirS), nitric oxide reductase (norB), and nitrous oxide reductase (nosZ) genes in seasonal sediment samples taken from the Acorus and Typha ponds of free surface flow constructed wetlands were investigated using quantitative polymerase chain reaction (Q-PCR) and quantitative reverse transcription PCR (Q-RT-PCR). Denitrifying gene copy numbers ($10^5-10^8$ genes $g^{-1}$ sediment) were found to be higher than transcript numbers-($10^3-10^7$ transcripts $g^{-1}$ sediment) of the Acorus and Typha ponds, in both seasons. Transcript numbers of the four functional genes were significantly higher for Typha sediments, in the warm than in the cold season, potentially indicating greater bacterial activity, during the relatively warm season than the cold season. In contrast, copy numbers and expression of denitrifying genes of Acorus did not provide a strong correlation between the different seasons.

• 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.40 no.3
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• pp.244-247
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• 2004

To investigate the effects of elevated $CO_2$ on the denitrifying bacterial community structure in a wetland soil, dynamics of bacterial community structure was explored in an artificial wetland ecosystem with one of three plant species (T. latifolia, S. lacustris, and 1. effusus) under two levels of $CO_2$(370 ppm or 740 ppm) after 110day incubation. For the analysis of bacterial community structure, functional genes such as nitrite reductase genes (nirS) were PCR-amplified followed by cloning of PCR products and screening by restriction fragment length polymorphism (RFLP). nirS gene fragments were amplified in all analyzed soil samples. Species richness estimated by the number of distinct phylotypes were 83 and 95 in the ambient $CO_2$ treatment and the elevated treatment, respectively. Two phylotypes (type 1 and type 2) were dominant in both of the treatments. Elevated $CO_2$ treatment increased species richness of denitrifying as well as changed a large proportion of denitrifier phylotypes compared to those of the ambient treatment. Overall, the data in this study suggested that the denitrifying communities in the wetland soil are diverse and that the richness of denitrifying bacterial community might be affected by elevated $CO_2$ treatment.

• Journal of Microbiology and Biotechnology
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• v.24 no.6
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• pp.843-853
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• 2014

In the context of artificial groundwater recharge, a reactive soil column at pilot-scale (4.5 m depth and 3 m in diameter) fed by treated wastewater was designed to evaluate soil filtration ability. Here, as a part of this project, the impact of treated wastewater filtration on soil bacterial communities and the soil's biological ability for wastewater treatment as well as the relevance of the use of multi-bioindicators were studied as a function of depth and time. Biomass; bacterial 16S rRNA gene diversity fingerprints; potential nitrifying, denitrifying, and sulfate-reducing activities; and functional gene (amo, nir, nar, and dsr) detection were analyzed to highlight the real and potential microbial activity and diversity within the soil column. These bioindicators show that topsoil (0 to 20 cm depth) was the more active and the more impacted by treated wastewater filtration. Nitrification was the main activity in the pilot. No sulfate-reducing activity or dsr genes were detected during the first 6 months of wastewater application. Denitrification was also absent, but genes of denitrifying bacteria were detected, suggesting that the denitrifying process may occur rapidly if adequate chemical conditions are favored within the soil column. Results also underline that a dry period (20 days without any wastewater supply) significantly impacted soil bacterial diversity, leading to a decrease of enzyme activities and biomass. Finally, our work shows that treated wastewater filtration leads to a modification of the bacterial genetic and functional structures in topsoil.

• Microbiology and Biotechnology Letters
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• v.48 no.1
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• pp.72-78
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• 2020

The dynamics of nitrogen compounds and RNA-based functional genes were characterized in the nitrification-denitrification coupling process. For the removal of residual ammonium, intermittent aeration was introduced in the denitrification reactor. N₂O production was not observed in both reactors. In both reactors, the nitrifying genes (achaeal-amoA, bacterial-amoA and hor) and denitrifying genes (narG, nirK, norB and nosZ) had a copy number of 3.92 × 10²-7.25 × 10⁵ and 2.85 × 10²-3.06 × 10⁴ per ng of DNA, respectively. These results suggest that denitrification and nitrification reactions occur in both the nitrification and denitrification reactors, respectively. Therefore, the coupling process is a promising one for the conversion of ammonium to nitrogen without generating N₂O.

• Journal of Microbiology and Biotechnology
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• v.33 no.4
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• pp.471-484
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• 2023

Compost is widely used as an organic additive to improve the bioremediation of diesel-contaminated soil. In this study, the effects of compost amendment on the remediation performance, functional genes, and bacterial community are evaluated during the bioremediation of diesel-contaminated soils with various ratios of compost (0-20%, w/w). The study reveals that the diesel removal efficiency, soil enzyme (dehydrogenase and urease) activity, soil CH₄ oxidation potential, and soil N₂O reduction potential have a positive correlation with the compost amendment (p < 0.05). The ratios of denitrifying genes (nosZI, cnorB and qnorB) to 16S rRNA genes each show a positive correlation with compost amendment, whereas the ratio of the CH₄-oxidizing gene (pmoA) to the 16S rRNA genes shows a negative correlation. Interestingly, the genera Acidibacter, Blastochloris, Erythrobacter, Hyphomicrobium, Marinobacter, Parvibaculum, Pseudoxanthomonas, and Terrimonas are strongly associated with diesel degradation, and have a strong positive correlation with soil CH₄ oxidation potential. Meanwhile, the genera Atopostipes, Bacillus, Halomonas, Oblitimonas, Pusillimonas, Truepera, and Wenahouziangella are found to be strongly associated with soil N₂O reduction potential. These results provide useful data for developing technologies that improve diesel removal efficiency while minimizing greenhouse gas emissions in the bioremediation process of diesel-contaminated soil.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.11
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• pp.1651-1658
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• 2013

Nitrate contamination in ground and surface water is an increasingly serious environmental problem and only a few bacterial strains have been identified that have the ability to remove nitrogen pollutants from wastewater under thermophilic conditions. We therefore isolated thermophilic facultative bacterial strains from wood chips that had been composted with swine manure under aerated high temperature conditions so as to identify strains with denitrifying ability. Nine different colonies were screened and 3 long rod-shaped bacterial strains designated as SG-01, SG-02, and SG-03 were selected. The strain SG-01 could be differentiated from SG-02 and SG-03 on the basis of the method that it used for sugar utilization. The 16S rRNA genes of this strain also had high sequence similarity with Geobacillus thermodenitrificans $465^T$ (99.6%). The optimal growth temperatures ($55^{\circ}C$), pH values (pH 7.0), and NaCl concentrations (1%) required for the growth of strain SG-01 were established. This strain reduced 1.18 mM nitrate and 1.45 mM nitrite in LB broth after 48 h of incubation. These results suggest that the G. thermodenitrificans SG-01 strain may be useful in the removal of nitrates and nitrites from wastewater generated as a result of livestock farming.

• Korean Journal of Microbiology
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• v.53 no.1
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• pp.58-60
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• 2017

Halioglobus sp. RR3-57 was isolated from a biofilter of a seawater recirculating aquaculture system and its complete genome sequence was obtained using the PacBio RS II platform. Two circular contigs were assembled and considered as a chromosome and a plasmid (size of 4,847,776 bp and 155,799 bp, and G+C content of 57.5% and 53.2%, respectively). Genomic analysis showed RR3-57 had 18 denitrification-related genes and an incomplete prophage.

• Journal of Korean Society of Environmental Engineers
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• v.28 no.7
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• pp.752-756
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• 2006

The independent anoxic reactor was introduced in biological aerated filters as the regulation of water quality requirement, especially total nitrogen, had been strengthened. The process studied in this work was upflow $Biobead^{(R)}$ process which was used commercial invented for removal of organic materials and nitrification. For the purpose of evaluating the independent anoxic reactor, PCR-DGGE, of the molecular biological methods, was performed. Two types of nitrite reductase genes were selected. One is nirS represented cytocrome $cd_1$ nitrite reductase gene and the other is nirK represented Cu-containing nitrite reductase gene. Denitrifier community in the independent anoxic reactor was analyzed with PCR-DGGE using these two denitrifying functional genes. As the result of the PCR, only nirS gene was detected between nirS and nirK. With the result of the DGGE, specific bands became strong, as the operating days were longer, nitrate loading rate was increased. otherwise those of the initial activated sludge showed various bands. In the consequence of the sequence of DGGE bands, various denitrifiers were sequenced in the initial activated sludge, while specific denitrifiers like alcaligenes faecalis were predominant in the anoxic reactor. Consequently, introduction of the independent anoxic reactor made it possible to achieve 96% denitrification efficiency, and was proper for the modification of BAF process.