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

• Korean Journal of Microbiology
• /
• v.40 no.2
• /
• pp.127-132
• /
• 2004

We isolated 50 strains from sludge of wastewater treatment aeration tank using selective medium for Thio-bacillus sp. by membrane filtration method. They were identified as Thiobacillus neapolitanus (7), T. tepidarius (4), T. dientrificans (5), T. versutus (23), T. intermedius (2) and T. perometabolis (9). We selected Thiobacillus versutus strain KT51, which had the highest removal efficiency (100％) of hydrogen sulfide and the highest removal efficiency (85％) of dimethyl sulfide for 30 min in screen test. Also Thiobacillus versutus strain KT81 had the highest removal efficiency (26％) of dimethyl disulfide for 30 min in screen test. In applification of lab-scale reactor (closed-biological treatment) using Thiobacillus versutus strain KT51, results were 99.8％ (＜0.02 ppm) removal efficiency of hydrogen sulfide for 15 min.

• Journal of Soil and Groundwater Environment
• /
• v.11 no.2
• /
• pp.38-44
• /
• 2006

Two sulfur-based column reactors inoculated with a bacterial consortium containing autotrophic denitrifiers were operated for 100 and 500 days, respectively and nitrate removal efficiency and the adaptation of microbial communities in the columns were monitored with column depths and time. For better understanding the adaptation phenomenon, molecular techniques including 16S rDNA sequencing and DGGE analysis were employed. Although both columns showed about 99% of nitrate removal efficiency heterotrophic denitrifiers such as Cenibacterium arsenioxidans and Geothrix fermentans were found to a significant portion at the initial stage of the 100-day reactor operation. However, as operation time increased, an autotrophic denitrifier Thiobacillus denitrificans became a dominant bacterial species throughout the column. A similar trend was also observed in the 500-day column. In addition, nitrate removal efficiencies were different with column depths and thus bacterial species with different metabolic activities were found at the corresponding depths. Especially, T. denitrificans was successfully adapted and colonized at the bottom parts of the columns where most nitrate was reduced.

• Korean Journal of Soil Science and Fertilizer
• /
• v.39 no.6
• /
• pp.409-414
• /
• 2006

Sulfur-oxidizing bacteria were enumerated and isolated from a steady-state anaerobic master culture reactor (MCR) operated for over six months under a semi-continuous mode and nitrate-limiting conditions using thiosulfate as an electron donor. Most are Gram-negative bacteria, which have sizes up to 12 m. Strains AD1 and AD2 were isolated from the plate count agar (PCA), and strains BD1 and BD2 from the solid thiosulfate/nitrate medium. Based on the morphological, physiological, FAME and 16S rDNA sequence analyses, the two dominant strains, AD1 and AD2, were identified as Paracoccus denitrificans and Paracoccus versutus (formerly Thiobacillus versutus), respectively. From the physiological results, glucose was assimilated by both strains AD1 and AD2. Heterotrophic growth of strains AD1 and AD2 could be a more efficient way of obtaining a greater amount of biomass for use as an inoculum. Even though facultative autotrophic bacteria grow under heterotrophic conditions, autotrophic denitrification would not be reduced.

• Journal of Soil and Groundwater Environment
• /
• v.14 no.3
• /
• pp.40-46
• /
• 2009

The applicability of a well-type autotrophic sulfur-oxidizing reactive barrier (L $\times$ W $\times$ D = $3m\;{\times}\;4\;m\;{\times}\;2\;m$) as a long-term treatment option for nitrate removal in groundwater was evaluated. Pilot-scale (L $\times$ W $\times$ D = $8m\;{\times}\;4\;m\;{\times}\;2\;m$) flow-tank experiments were conducted to examine remedial efficacy of the well-type reactive barrier. A total of 80 kg sulfur granules as an electron donor and Thiobacillus denitrificans as an active bacterial species were prepared. Thiobacillus denitrificans was successfully colonized on the surface of the sulfur granules and the microflora transformed nitrate with removal efficiency of ~12% (0.07 mM) for 11 days, ~24% (1.3 mM) for 18 days, ~45% (2.4 mM) for 32 days, and ~52% (2.8 mM) for 60 days. Sulfur granules attached to Thiobacillus denitrificans were used to construct the well-type reactive barrier comprising three discrete barriers installed at 1-m interval downstream. Average initial nitrate concentrations were 181 mg/L for the first 28 days and 281 mg/L for the next 14 days. For the 181 mg/L (2.9 mM) plume, nitrate concentrations decreased by ~2% (0.06 mM), ~9% (0.27 mM), and ~15% (0.44 mM) after $1^{st}$, $2^{nd}$, and $3^{rd}$ barriers, respectively. For the 281 mg/L (4.5 mM) plume, nitrate concentrations decreased by ~1% (0.02 mM), ~6% (0.27 mM), and ~8% (0.37 mM) after $1^{st}$, $2^{nd}$, and $3^{rd}$ barriers, respectively. Nitrate plume was flowed through the flow-tank for 49 days by supplying $1.24\;m^3/d$ of nitrate solution. During nitrate treatment, flow velocity (0.44 m/d), pH (6.7 to 8.3), and DO (0.9~2.8 mg/L) showed little variations. Incomplete destruction of nitrate plume was attributed to the lack of retention time, rarely transverse dispersion, and inhibiting the activity of denitrification enzymes caused by relatively high DO concentrations. For field applications, it should be considered increments of retention time, modification of well placements, and intrinsic DO concentration.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
• /
• 2000.11a
• /
• pp.209-212
• /
• 2000

As a bench-scale study, transformation of nitrate to nitrogen gas under anoxic condition was determined by using autotrophic denitrifiers containing Thiobacillus denitrificans and elemental sulfur as an electron donor. The research objective is to measure the basic kinetic parameters of autotrophic denitrification reaction on the removal efficiency of nitrate. The results showed that nitrate was almost completely transformed to nitrite in the first 4 days of column operation. After 2 days of accumulation of nitrite, its concentration slowly decreased and the compound was detected less than 0.5 mg/L in 14 days. In the experiment, sulfate concentration in the effluent was the 70~90 mg-S/L and the pH was maintained around pH 7.5. When nitrate concentration of bank filtrate in the real field is considered, this sulfate concentration seems to be acceptable. At 17 cm from the bottom of the column, the effluent showed the highest nitrite concentration, and nitrate concentration decreased rapidly to the Point of 33 cm from the bottom. The results suggest that an appropriate thickness of permeable reactive barriers is about 30 cm.

• Proceedings of the Korea Water Resources Association Conference
• /
• 2007.05a
• /
• pp.46-52
• /
• 2007

As flue gas desulfurization (FGD) wastewater contains high concentrations of nitrate and is very low in organic carbon, the feasibility of nitrate removal by autotrophic denitrification using Thiobacillus denitrificans was studied. This autotrophic bacteria oxidizes elemental sulfur to sulfate while reducing nitrate to elemental nitrogen gas, thereby eliminating the need for addition of organic compounds such as methanol. Owing to the unusually high concentrations of dissolved salts $(Ca^{2+},\;Mg^{2+},\;Na^+,\;K^+,\;B^+,\;SO_4^{2-},\;Cl^-,\;F^-,)$ in the FGD wastewater, extensive laboratory-scale and pilot-scale tests were carried out in sulfur-limestone reactors (1) to determine the effect of salinity on autotrophic denitrification, (2) to evaluate the use of limestone for pH control and as source of inorganic carbon for microbial growth, and, (3) to find the optimum environmental and operational conditions for autotrophic denitrification of FGD wastewater. The experimental results demonstrated that (1) autotrophic denitrification is not inhibited up to 1.8 mol total dissolved salt content; (2) inorganic carbon and inorganic phosphorus must be present in sufficiently high concentrations; (3) limestone can supply effective buffering capacity and inorganic carbon; (4) the high calcium concentration may interfere with pH control, phosphorus solubility and limestone dissolution, hence requiring pretreatment of the FGD wastewater; and, 5) under optimum conditions, complete autotrophic denitrification of FGD wastewater was obtained in a sulfur-limestone packed bed reactor with a sulfur:limestone volume ratio of 2:1 for volumetric loading rates up to 400g $NO_{3^-}N/m^3.d$. The interesting interactions between autotrophic denitrification, pH, alkalinity, and the unusually high calcium and boron content of the FGD wastewater are highlighted. The engineering significance of the results is discussed.