• Journal of Soil and Groundwater Environment
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• v.27 no.3
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• pp.41-49
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• 2022

This study utilized citric acid as a floating agent in biological denitrification process and assessed its role under different carbon supplying conditions. Several microcosm tests including citric acid active (CAA), precipitating tablet release active (PTRA) and floating tablet release active (FTRA) were conducted to evaluate nitrate denitrification efficacy. In CAA reactors, nitrate removal was accompanied by the formation of denitrification by-products such as nitrite and nitrous oxide, with the extent of nitrate removal being proportional to citric acid concentration. These results suggest that citric acid induced heterotrophic biological denitrification. PTRA reactor that incorporated CAA and the same electron donor showed a similar denitrification efficiency to CAA reactor. FTRA reactor, which contained the same amount of fumarate as PTRA, enhanced denitrification by 7% as compared to the PTRA reactor. The overall results of this work indicate that surplus citric acid can be efficiently utilized in heterotrophic denitrification.

• Journal of Microbiology and Biotechnology
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• v.18 no.11
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• pp.1826-1835
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• 2008

An autotrophic denitrification reactor (ADR-l) and a heterotrophic denitrification reactor (HDR-2) were operated to remove nitrate and nitrite in an anoxic environment in raw sewage. The $NO_3$-N removal rate of ADR-l was shown to range from 52.8% to 78.7%, which was higher than the $NO_3$-N removal rate of HDR-2. Specific denitrification rates (SDNR) of ADR-l and HDR-2 were 3.0 to 4.0 and 1.1 to $1.2\;mgNO_3$-N/gVSS/h, respectively. From results of restriction fragment length polymorphism (RFLP) of the 16S rRNA gene, Aquaspirillum metamorphum, Alcaligenes defragrans, and Azoarcus sp. were $\beta$-Proteobacteria that are affiliated with denitritying bacteria in the ADR-l. Specifically, Thiobacillus denitrificans was detected as an autotrophic denitrification bacteria. In HDR-2, the $\beta$-Proteobacteria such as Denitritying-Fe-oxidizing bacteria, Alcaligenes defragrans, Acidovorax sp., Azoarcus denitrificans, and Aquaspirillum metamorphum were the main bacteria related to denitrifying bacteria. The $\beta$-and $\alpha$-Proteobacteria were the important bacterial groups in ADR-l, whereas the $\beta$-Proteobacteria were the main bacterial group in HDR-2 based on results of fluorescent in situ hybridization (FISH). The number of Thiobacillus denitrificans increased in ADR-l during the operation period but not in HRD-2. Overall, the data presented here demonstrate that many heterotrophic denitritying bacteria coexisted with autotrophic denitrifying bacteria such as Thiobacillus denitrificans for nitrate removal in ADR-l. On the other hand, only heterotrophic denitritying bacteria were identified as dominant bacterial groups in HDR-2. Our research may provide a foundation for the complete nitrate removal in raw sewage of low-COD concentration under anoxic condition without any external organic carbon or the requirement of post-treatment.

• Journal of Microbiology and Biotechnology
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• v.22 no.8
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• pp.1155-1160
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• 2012

Sulfur-utilizing autotrophic denitrification relies on an inorganic carbon source to reduce the nitrate by producing sulfuric acid as an end product and can be used for the treatment of wastewaters containing high levels of nitrates. In this study, sulfur-denitrifying bacteria were used in anoxic batch tests with sulfur as the electron donor and nitrate as the electron acceptor. Various medium components were tested under different conditions. Sulfur denitrification can drop the medium pH by producing acid, thus stopping the process half way. To control this mechanism, a 2:1 ratio of sulfur to oyster shell powder was used. Oyster shell powder addition to a sulfur-denitrifying reactor completely removed the nitrate. Using 50, 100, and 200 g of sulfur particles, reaction rate constants of 5.33, 6.29, and $7.96mg^{1/2}/l^{1/2}{\cdot}h$ were obtained, respectively; and using 200 g of sulfur particles showed the highest nitrate removal rates. For different sulfur particle sizes ranging from small (0.85-2.0 mm), medium (2.0-4.0 mm), and large (4.0-4.75 mm), reaction rate constants of 31.56, 10.88, and $6.23mg^{1/2}/l^{1/2}{\cdot}h$ were calculated. The fastest nitrate removal rate was observed for the smallest particle size. Addition of chemical oxygen demand (COD), methanol as the external carbon source, with the autotrophic denitrification in sufficiently alkaline conditions, created a balance between heterotrophic denitrification (which raises the pH) and sulfur-utilizing autotrophic denitrification, which lowers the pH.

• 한국환경농학회:학술대회논문집
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• 2011.07a
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• pp.296-296
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• 2011

• Environmental Engineering Research
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• v.24 no.3
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• pp.443-448
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• 2019

The present study was conducted to compare the voltage generation in two-chamber microbial fuel cells (MFCs) with a biocathode where nitrate and oxygen are used as a terminal electron acceptors (TEA) and to investigate the nitrogen removal and the electrochemical characteristics depending on the separators of the MFCs for denitrification. The maximum power density in a biocathode MFC using an anion exchange membrane (AEM) was approximately 40% lower with the use of nitrate as a TEA than when using oxygen. The MFC for denitrification using an AEM allows acetate ($CH_3COO^-$) as a substrate and nitrate ($NO_3{^-}$) as a TEA to be transported to the opposite sides of the chamber through the AEM. Therefore, heterotrophic denitrification and electrochemical denitrification occurred simultaneously at the anode and the cathode, resulting in a higher COD and nitrate removal rate and a lower maximum power density. The MFC for the denitrification using a cation exchange membrane (CEM) does not allow the transport of acetate and nitrate. Therefore, as oxidation of organics and electrochemical denitrification occurred at the anode and at the cathode, respectively, the MFC using a CEM showed a higher coulomb efficiency, a lower COD and nitrate removal rate in comparison with the MFC using an AEM.

• Korean Journal of Microbiology
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• v.52 no.2
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• pp.166-174
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• 2016

In order to find an efficient bacterial strain that can carry out nitrification and denitrification simultaneously, we isolated many heterotrophic nitrifying bacteria from wastewater treatment plant. One of isolates NS13 showed high removal rate of ammonium and was identified as Alcaligenes faecalis by analysis of its 16S rDNA sequence, carbon source utilization and fatty acids composition. This bacterium could remove over 99% of ammonium in a heterotrophic medium containing 140 mg/L of ammonium at pH 6-9, $25-37^{\circ}C$ and 0-4% of salt concentrations within 2 days. It showed even higher ammonium removal at higher initial ammonium concentration in the medium. A. faecalis NS13 could also reduce nitrate and nitrous oxide by nitrate reductase and nitrous oxide reductase, respectively, which was confirmed by detection of nitrate reductase gene, napA, and nitrous oxide reducase gene, nosZ, by PCR. One of metabolic intermediate of denitrification, $N_2O$ was detected from headspace of bacterial culture. Based on analysis of all nitrogen compounds in the bacterial culture, 42.8% of initial nitrogen seemed to be lost as nitrogen gas, and 46.4% of nitrogen was assimilated into bacterial biomass which can be removed as sludge in treatment processes. This bacterium was speculated to perform heterotrophic nitrification and aerobic denitrification simultaneously, and may be utilized for N removal in wastewater treatment processes.

• Journal of Soil and Groundwater Environment
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• v.15 no.4
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• pp.30-38
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• 2010

This study was conducted to determine the potential applicability of slowly released molasses (SRM) to treat nitratecontaminated groundwater. SRM was made by dispersing molasses in hydroxy propyl methyl cellulose-silicamicrocrystalline cellulose matrix. Column test indicated that SRM could continuously release molasses with slowly decreasing release rates of $64.6mg-COD/L{\cdot}h$ up to 65 hrs, $12.1mg-COD/L{\cdot}h$ up to 215 hrs, and $4.4mg-COD/L{\cdot}h$up to 361 hrs. A batch test using an isolated indigenous heterotrophic denitrifier Pseudomonas sp. KY1 having nitrite reductase (nirK) and liquid molasses demonstrated that the bacterium decreased 100 mg-N/L of nitrate to less than 10 mg-N/L at the C/N ratio of 10/1 in 48 hours. In a Pseudomonas sp. KY1-attached Ottawa sand column which continuously received molasses from a SRM-containing reservoir, the bacterium successfully removed nitrate from 20 mg-N/L to 3 mg-N/L during the 361 hours of column operation. The results showed the possibility that SRM can be used as a reliable, longterm extra carbon source for indigenous heterotrophic denitrifiers.

• Journal of Environmental Science International
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• v.21 no.8
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• pp.989-996
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• 2012

Nitrate contamination of water environments can create serious problems such as eutrophication of rivers. Conventional biological processes for nitrate removal by heterotrophic denitrification often need additional organic substrates as carbon sources and electron donors. We tried to accelerate biological denitrification by using bioelectrochemical reactor (BER) in which electrode works as an electron donor. Denitrification activity of 8 environmental samples from various sediments, soils, groundwaters, and sludges were tested to establish an efficient enrichment culture for BER. The established enrichment culture from a soil sample showed stable denitrification activity without any nitrite accumulation. Microbial community analysis by using PCR-DGGE method revealed that dominant denitrifiers in the enrichment culture were Pantoea sp., Cronobacter sakazakii, and Castellaniella defragrans. Denitrification rate ($0.08kg/m^3{\cdot}day$) of the enrichment culture in BER with electrode poised at -0.5 V (vs Ag/AgCl) was higher than that ($2.1{\times}10^{-2}kg/m^3{\cdot}day$) of BER without any poised potential. This results suggested that biological denitrification would be improved by supplying potential throughout electrode in BER. Further research using BER without any organic substrate addition is needed to apply this system for bioremediation of water and wastewater contaminated by nitrate.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 2004.04a
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• pp.77-82
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• 2004

Quantifying rates of microbial processes under subsurface conditions is difficult, and is most commonly approximated by laboratory studies using aquifer materials. In this study a single-well, 'push-pull' test method is adapted for the in situ determination of denitrification rates in groundwater aquifers. The rates of stepwise reduction of nitrate to nitrite, nitrous oxide, and molecular nitrogen were determined by performing a series of push-pull tests at an experimental well field of Korea University. A single Transport Test, one Biostimulation Test, and four Activity Tests were conducted for this study. Transport tests are conducted to evaluate the mobility of solutes used in subsequent tests. These included bromide (a conservative tracer), fumarate (a carbon and/or source), and nitrate (an electron acceptor). At this site, extraction phase breakthrough curves for all solutes were similar, indicating apparent conservative transport of the solutes prior to biostimulation. Biostimulation tests were conducted to stimulate the activity of indigenous heterotrophic denitrifyinc microorganisms. Biostimulation was detected by the simultaneous production of carbon dioxide and nitrite after each injection. Activity tests were conducted to quantify rates of nitrate, nitrite, and nitrous oxide reduction. Estimated zero-order degradation rates decreased in the order nitrate '||'&'||'gt; nitrite '||'&'||'gt; nitrous oxide. The series of push-pull tests developed and field tested in this study should prove useful for conducting rapid, low-cost feasibi1ity assessments for in situ denitrification in nitrate-contaminated aquifers.

• Journal of Korean Society of Environmental Engineers
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• v.35 no.9
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• pp.654-659
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• 2013

This study was conducted to identify an optimal ratio of carbon to nitrogen (C/N ratio) for denitrification of nitrate using molasses as an external carbon source. A series of batch and column tests was conducted using an indigenous bacterium Pseudomonas sp. KY1 isolated from a nitrate-contaminated soil. For the initial nitrate-nitrogen concentration of 100 mg-N/L, batch test results indicated that C/N ratio of 3/1 was the optimal ratio with a relatively high pseudo-first-order reaction constant of $0.0263hr^{-1}$. At C/N ratio of 3/1, more than 80% of nitrate-nitrogen concentration of 100 mg-N/L was removed in 100 hrs. Results of column tests with a flow velocity of 0.3 mL/min also indicated that the C/N ratio of 3/1 was optimal for denitrification with minimizing remaining molasses concentrations. After 172 hrs of column operation (35 pore volumes) with an influent nitrate-nitrogen concentration of 100 mg-N/L, the effluent met the drinking water standard (i.e., 10 mg $NO_3$-N/L).