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Evaluation of Growth Inhibition for Microcystis aeruginosa with Ultrasonic Irradiation Time (초음파 조사시간에 따른 Microcystis aeruginosa의 성장억제 평가)

  • Kang, Eun Byeol;Joo, Jin Chul;Jang, So Ye;Go, Hyeon Woo;Park, Jung Su;Jeong, Moo Il;Lee, Dong Ho
    • Ecology and Resilient Infrastructure
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    • v.9 no.3
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    • pp.183-193
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    • 2022
  • The growth inhibitory effect of Microcystis aeruginosa according to the ultrasonic irradiation time was evaluated using a large algae sample volume (10 L) for various ultrasonic irradiation times (0.5, 1, 1.5, 2, 2.5 and 3 hr) at a laboratory scale. Based on the analysis of Chl-a and cell number of M. aerginosa, algae growth inhibition was observed with the decrease in Chl-a and cell number in all experimental groups after the ultrasonic irradiation. For the experimental group (T_B, T_C, T_D) with an ultrasonic irradiation time of less than 2 hours, rapid regrowth of algae was observed after growth inhibition, but the experimental group (T_E, T_F, T_G) with an irradiation time of more than 2 hours successfully inhibited algal growth lasting one or two more days. Based on the comparison of the recovery time to initial cell number the experimental group (T_B, T_C, T_D) took less than 20 days whereas the experimental group (T_E, T_F, T_G) took about 30 days. Correspondingly, the experimental group showed a high first order decay rate (𝜅) in proportion to the ultrasonic irradiation time during the growth inhibition period. Additionally, the specific growth rates (𝜇) during regrowth in the experimental group with irradiation time of more than 2 hours were relatively low compared to those in the experimental group with less than 2 hours. Therefore, ultrasonic irradiation for more than 2 hours is required for long-term (30 days) inhibition of algal growth in stagnant waters. However, the appropriate ultrasonic irradiation time for algae growth inhibition should be determined according to various field conditions such as the volume of stagnant water, water depth, flow rate, algae concentration, etc. Finally, damages to the algal cell surface and cell membrane were clearly observed, and both destruction and disturbance of gas vesicles of M. aeruginosa in the experimental group were discovered, indicating the growth inhibitory effect of Microcystis aeruginosa according to the ultrasonic irradiation time was confirmed.

Influence of a chemical additive on the reduction of highly concentrated ammonium nitrogen(NH4+-N) in pig wastewater (양돈 폐수로부터 고농도 암모니아성 질소의 감소를 위한 화학적 첨가제의 영향)

  • Su Ho Bae;Eun Kim;Keon Sang Ryoo
    • Korean Journal of Environmental Biology
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    • v.40 no.3
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    • pp.267-274
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    • 2022
  • Excess nitrogen (N) flowing from livestock manure to water systems poses a serious threat to the natural environment. Thus, livestock wastewater management has recently drawn attention to this related field. This study first attempted to obtain the optimal conditions for the further volatilization of NH3 gas generated from pig wastewater by adjusting the amount of injected magnesia (MgO). At 0.8 wt.% of MgO (by pig wastewater weight), the volatility rate of NH3 increased to 75.5% after a day of aeration compared to untreated samples (pig wastewater itself). This phenomenon was attributed to increases in the pH of pig wastewater as MgO dissolved in it, increasing the volatilization efficiency of NH3. The initial pH of pig wastewater was 8.4, and the pH was 9.2 when MgO was added up to 0.8 wt.%. Second, the residual ammonia nitrogen (NH4+-N) in pig wastewater was removed by precipitation in the form of struvite (NH4MgPO4·6H2O) by adjusting the pH after adding MgO and H3PO4. Struvite produced in the pig wastewater was identified by field emission scanning electron microscopy (FE-SEM) and X-ray diffraction (XRD) analysis. White precipitates began to form at pH 6, and the higher the pH, the lower the concentration of NH4+-N in pig wastewater. Of the total 86.1% of NH4+-N removed, 62.4% was achieved at pH 6, which was the highest removal rate. Furthermore, how struvite changes with pH was investigated. Under conditions of pH 11 or higher, the synthesized struvite was completely decomposed. The yield of struvite in the precipitate was determined to be between 68% and 84% through a variety of analyses.

Selection and appropriate culture conditions of antagonistic bacterium Bacillus altitudinis HC7 against button mushroom cobweb disease caused by Cladobotryum mycophilum (양송이버섯 솜털곰팡이병균(Cladobotryum mycophilum)에 대한 길항미생물 Bacillus altitudinis HC7의 선발 및 적정 배양조건)

  • Chan-Jung Lee;Hye-Sung Park;Seong-Yeon Jo;Gi-Hong An;Ja-Yun Kim;Kang-Hyo Lee
    • Journal of Mushroom
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    • v.22 no.2
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    • pp.60-66
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    • 2024
  • This study was conducted to selection and investigate appropriate conditions for mass production of antagonistic microbes to control cobweb disease caused by Cladobotryum mycophilum. A grampositive bacterium was isolated from spent substrate of Agaricus bisporus and showed significant antagonistic activity against Cladobotryum mycophilum. The bacterium was identified as Bacillus altitudinis. based on the cultural, biochemical and physiological characteristics, and 16S rRNA sequence. The isolate is saprophytic, but not parasitic nor pathogenic to cultivated mushroom whereas it showed strong inhibitory effects against C. mycophilum cells in vitro. The control efficacy of B. altitudinis HC7 against cobweb disease of C. mycophilum was up to 78.2% on Agaricus bisporus. The suppressive bacterium may be useful for the development of biocontrol system. To define the appropriate conditions for the mass production of the Bacillus altitudinis HC7, we have investigated appropriate culture conditions and effects of various nutrient source on the bacterial growth. The appropriate initial pH and temperature were determined as pH 6.0 and 30℃, respectively. The appropriate concentration of medium elements for the growth of pathogen inhibitor bacterium(Bacillus altitudinis HC7) was determined as follows: 3.0% soluble startch, 10% soytone, 1.0% (NH4)2HPO4, 1.0 mmol KCl, and 0.5% L-asparagine.

Establishment of Test Conditions and Interlaboratory Comparison Study of Neuro-2a Assay for Saxitoxin Detection (Saxitoxin 검출을 위한 Neuro-2a 시험법 조건 확립 및 실험실 간 변동성 비교 연구)

  • Youngjin Kim;Jooree Seo;Jun Kim;Jeong-In Park;Jong Hee Kim;Hyun Park;Young-Seok Han;Youn-Jung Kim
    • Journal of Marine Life Science
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    • v.9 no.1
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    • pp.9-21
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    • 2024
  • Paralytic shellfish poisoning (PSP) including Saxitoxin (STX) is caused by harmful algae, and poisoning occurs when the contaminated seafood is consumed. The mouse bioassay (MBA), a standard test method for detecting PSP, is being sanctioned in many countries due to its low detection limit and the animal concerns. An alternative to the MBA is the Neuro-2a cell-based assay. This study aimed to establish various test conditions for Neuro-2a assay, including cell density, culture conditions, and STX treatment conditions, to suit the domestic laboratory environment. As a result, the initial cell density was set to 40,000 cells/well and the incubation time to 24 hours. Additionally, the concentration of Ouabain and Veratridine (O/V) was set to 500/50 μM, at which most cells died. In this study, we identified eight concentrations of STX, ranging from 368 to 47,056 fg/μl, which produced an S-shaped dose-response curve when treated with O/V. Through inter-laboratory variability comparison of the Neuro-2a assay, we established five Quality Control Criteria to verify the appropriateness of the experiments and six Data Criteria (Top and Bottom OD, EC50, EC20, Hill slop, and R2 of graph) to determine the reliability of the experimental data. The Neuro-2a assay conducted under the established conditions showed an EC50 value of approximately 1,800~3,500 fg/μl. The intra- & inter-lab variability comparison results showed that the coefficients of variation (CVs) for the Quality Control and Data values ranged from 1.98% to 29.15%, confirming the reproducibility of the experiments. This study presented Quality Control Criteria and Data Criteria to assess the appropriateness of the experiments and confirmed the excellent repeatability and reproducibility of the Neuro-2a assay. To apply the Neuro-2a assay as an alternative method for detecting PSP in domestic seafood, it is essential to establish a toxin extraction method from seafood and toxin quantification methods, and perform correlation analysis with MBA and instrumental analysis methods.

Analysis of the Reduction Effect of Combined Treatment with UV-C and Organic Acid to Reduce Aspergillus ochraceus and Rhodotorula mucilaginosa Contamination (Aspergillus ochraceus와 Rhodotorula mucilaginosa 저감을 위한 자외선과 유기산 복합처리 효과 분석)

  • Eun-Seon Lee;Jong-Hui Kim;Bu-Min Kim;Mi-Hwa Oh
    • Journal of Food Hygiene and Safety
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    • v.39 no.1
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    • pp.54-60
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    • 2024
  • This study investigated the effectiveness of using pathogens and aqueous acids to reduce the Aspergillus ochraceus and Rhodotorula mucilaginosa contamination in livestock production environments. For this study, 1 mL of each bacterial suspension (107-108 spores/mL) was inoculated on a knife surface, dried at 37℃, and used under each treatment condition. First, to investigate the effect of organic acids, acetic, lactic, and citric acids were used. Subsequently, to select the appropriate concentration, they were prepared at concentrations of 0.5, 1, 2, 3, 4, and 5%, respectively. Accordingly, to further maximize the effect of organic acid treatment, we combined the treatment with ultraviolet light. The two strains showed a significant difference (P<0.05) compared to the initial strain, with a greater than 90% decrease in the concentrations of all organic acids. Consequently, acetic and lactic acids decreased by approximately 5 and 2 log colony forming unit (CFU)/cm2, respectively, when treated with ultraviolet light (360 mJ/cm2); however, citric acid decreased by less than 1 log CFU/cm2. However, when manufactured with 4% acetic acid, a severe malodor was emitted, making it difficult for workers to use it in a production environment. Accordingly, the optimal treatment conditions for organic acid and ultraviolet light for application were selected as follows: immersion in a 4% lactic acid solution for 1 minute and then, sterilization with ultraviolet light at 360 mJ/cm2. Finally, when a pork meat sample was cut with a knife that was finally washed with lactic acid and treated with ultraviolet light, the low level of inoculum transferred from the cleaned knife to the surface of the sample was not detected. In conclusion, using this established method can prevent cross-contamination of the surface of the meat during processing.

The Factors Controlling the Formation of Spring Population of Acartia hongi (Copepoda: Calanoida) in Incheon Coastal Water, Korea (인천 연안에서 요각류 Acartia hongi 춘계 개체군 형성의 영향 요인)

  • Yoo J.K.;Youn S.H.;Choi J.K.
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.11 no.3
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    • pp.108-116
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    • 2006
  • To investigate the factors controlling the spring population of Acartia hongi, egg production, hatching time of egg and predation pressure were measured. Egg production was maintained the superior position between winter and spring. Egg production was positively correlated with not only water temperature when water temperature was below $7^{\circ}C$ but also chlorophyll-a concentration when it was from $7^{\circ}C$ to $21^{\circ}C$. A regressive equation of development time$(D_e,\;day)$ of eggs derived from water temperature$(T,\;^{\circ}C)$ was obtained as $D_e=18.9(T-0.4)^{-1.0}$, showing longer development time at water temperature below $5^{\circ}C$. In Cross Correlation Analysis(CCA) to examine the time-lag relationship among abundances of developmental stages of A. hongi, egg production rate calculated by multiplying population egg production rate by hatching time showed more significant correlation with nauplii abundance than population egg production rate. Therefore, it suggests that hatching time is also recognized by a factor controlling formation of population and especially, in winter, high abundance of egg derived from high daily egg production rate and delayed hatch by low water temperature become the origin of initial spring population of nauplii. Egg predation by Noctiluca scintillans, suggesting a negative factor in formation of A. hongi spring population, was observed. During spring, A. hongi eggs were found in $2.9\sim21.1%$ of individuals of N. scintillans. It was deduced that $1.2\sim49.5%$ of the eggs produced by A. hongi was preyed on by N. scintillans. In conclusion, the factors controlling spring population of A. hongi were regarded as high egg production by winter generation, the delayed development time of egg by low temperature, and egg predation of N. scintillans.

Bloom-forming Cyanobacteria in Yongdam Lake (1) Nutrient limitation in a Laboratory Strain of a Nitrogen-fixing Cyanobacterium, Anabaena spiroides v. crassa (용담호 녹조현상의 원인 남세균 연구 (1) 질소고정 남세균 Anabaena spiroides v. crassa 종주와 영양염 제한)

  • Park, Jong-Woo;Kim, Young-Geel;Heo, Woo-Myung;Kim, Bom-Chul;Yih, Won-Ho
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.11 no.4
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    • pp.158-164
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    • 2006
  • Yongdam Lake is the fifth largest artificial lake in Korea newly formed by the first impounding the Yongdam Multi-purpose Dam on December, 2002. Yongdam Lake, with her total water storage of 820 million M/T, is located at the roof-top region of the streams flowing into the just-constructed new Saemankeum Lake. Seasonal succession of phytoplakton in Yongdam Lake might affect cyanobacterial blooms in Saemankeum Lake by inoculating seasonal dominants. During 2002-2003 when the first impounding after the construction of Yongdam Multi-purpose Dam was still undergoing, summer cyanobacterial blooms by Anabaena, Microcystis, and Aphanizomenon were observed. Among these three, filamentous Anabaena is well known to have its species with $N_2-fixing$ ability and special cells such as heterocysts and akinetes as well as the vegetative cells. We established a clonal culture of Anabaena spiroides v. crasse (KNU-YD0310) from the live water samples collected at the bloom site of Yongdam Lake. The N- and P-nutrient requirement of the KNU-YD0310 was explored by the experimental cultivation of the laboratory strain. Ratio of heterocysts to vegetative cells increased as N-deficiency extended with its maximum at $N_2-fixing$ condition. The strain KNU-YD0310 exhibited considerable growth under N-limiting conditions while its growth was proportional to the initial phosphate-P concentration under P-deficient conditions. Under P-limiting conditions akinete density increased, which could be interpreted as an adaptation strategy to survive severe environment by transforming into resting stage. The above eco-physiological characteristics of Anabaena spiroides v. crassa might be useful as an ecological criterion in controlling cyanobacterial blooms at Shaemankeum Lake in near future.

A Study on the Conversion to Feed Stuff from Cellulosic Biomass (섬유질자원(纖維質資源)의 사료(飼料) 전환(轉換))

  • Lee, Ke-Ho;Sung, Chang-Geun;Chung, Kyu-Ok
    • Applied Biological Chemistry
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    • v.27
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    • pp.29-46
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    • 1984
  • To utilize several species of hard wood as raw materials of feed products, fermentation characteristics of cellulosic substrates to single cell protein was investigated, and results were summarized as follows. Among the microorganisms investigated, Tricoderma viride was selected as one of the most cellulolytic. Mixed culture of fungi did not show a synergistic effect on cellulose degradation. When the fungi were cultured at $28^{\circ}C$ for 7 days in a medium containing wheat bran 25 g, cellulose 0.25 g, proteose peptone 0.025 g and tween 800.025 g, cellulotic activities on carboxy methyl cellulose and filter paper reached maximum at 12 hr. The alkali treatment resulted in increased degradation of substrate from 13 to 18% when treated with enzymes for 12h, and reducing sugar formation increased with decreased size of substrates. Glucose was a very good feedback inhibitor of the enzyme from T.viride than that of xylose. When the substrate was rehydrolyzed, hydrolysis rate was 31% to reducing sugars within 12 hr. Quantative anlysis with HPLC showed the ratio of glucose to xylose in sugar syrups as 1.77 to 1. For the purpose of producing cellulosic-single cell protein from the sawdust of mulberry tree, 15 strains of xylose-assimilating yeast were isolated from 42 samples of rotten woods and compost soils and examined for their ability to utilize xylose. Then three strains were selected by their strong xylose-assimilating activities. The cultivative condition, the growth characteristics, and protein and nucleic acid productivities of three strains were investigated. The results obtained were, 1. Wood hydrolysate of mulberry tree was assimilated by 5 strains of CHS-2, CHS-3, ST-40, CHS-12 and CHS-13. 2. The optimum initial pH and temperature for the growth of strain CHS-13 were 4.4 and $30^{\circ}C$. 3. The specific growth rate of strain CHS-13 was $0.23h^{-1}$ and generation time was 3.01 hrs at the optimum condition. 4. CHS-13 strain assimilated 81 % of sugar in wood hydrolysate. 5. CHS-13 strain was identified as Candida guilliermondii var. guilliermondii 6. When the CHS-13 strain was cultured in the wood hydrolysate containing yeast extract, L-protein content was increased with yeast extract concentration. 7. The L-protein and nucleic acid yields from wood hydrolysate were 0.73 mg/ml and $4.92{\times}10^{-2}\;mg/ml$ respectively. 8. An optimal nucleic acid content of CHS-13 strain was observed in the medium containing 0.2% of yeast extract.

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Abundance of Harmful Algae, Cochlodinium polykrikoides, Gyrodinium impudicum and Gymnodinium catenatum in the Coastal Area of South Sea of Korea and Their Effects of Temperature, Salinity, Irradiance and Nutrient on the Growth in Culture (남해안 연안에서 적조생물, Cochlodinium polykikoides, Gyrodinium impudicum, Gymnodinium catenatum의 출현상황과 온도, 염분, 조도 및 영양염류에 따른 성장특성)

  • LEE Chang Kyu;KIM Hyung Chul;LEE Sam-Geun;JUNG Chang Su;KIM Hak Gyoon;LIM Wol Ae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.34 no.5
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    • pp.536-544
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    • 2001
  • Three harmful algal bloom species with similar morphology, Cochlodinium polykrikoides, Gyodinium impudicum and Gymodinium catenatum have damaged to aquatic animals or human health by either making massive blooms or intoxication of shellfishes in a food chain. Eco-physiological and hydrodynamic studies on the harmful algae offer useful informations in the understanding their bloom mechanism by giving promising data for the prediction and modelling of harmful algal blooms event. Thus, we studied the abundance of these species in the coastal area of South Sea of Korea and their effects of temperature, salinity, irradiance and nutrient on the growth for the isolates. The timing for initial appearance of the three species around the coastal area of Namhaedo, Narodo and Wando was between Bate July and late August in 1999 when water temperature ranged from $22.8^{\circ}C\;to\;26.5^{\circ}C$ Vegetative cells of C. polykrikoides and G. impudicum were abundant until late September when water temperature had been dropped to less than $23^{\circ}C$. By contrast, vegetative cell of G. catenatum disappeared before early September, showing shorter period of abundance than the other two species in the South Sea. Both G. impudicum and G. catenatum revealed comparatively low density with a maximal cell density of 3,460 cells/L and 440 cells/L, respectively without making any bloom, while C. polykrikoides made massive blooms with a maximal cell density more than $40\times10^6$cells/L, The three species showed a better growth at the relatively higher water temperature ranging from 22 to $28^{\circ}C$ with their maximal growth rate at $25^{\circ}C$ in culture, which almost corresponded with the water temperature during the outbreak of C. polykrikoides in the coastal area of South Sea. Also, they all showed a relatively higher growth at the salinity from 30 to $35\%$. Specially, G. impudicum showed the euryhalic characteristics among the species, On the other hand, growth rate of G. catenatum decreased sharply with the increase of water temperature at the experimental ranges more than $35\%$. The higher of light intensities showed the better growth rates for the three species, Moreover, C. polykrikoides and G. impudirum continued their exponential growth even at 7,500 lux, the highest level of light intensity in the experiment, Therefore, It is assumed that C. polykrikoides has a physiological capability to adapt and utilize higher irradiance resulting in the higher growth rate without any photo inhibition response at the sea surface where there is usually strong irradiance during its blooming season. Although C. poiykikoides and G. impudicum continued their linear growth with the increase of nitrate ($NO_3^-$) and ammonium ($NH_4^-$) concentrations at less than the $40{\mu}M$, they didn't show any significant differences in growth rates with the increase of nitrate and ammonium concentrations at more than $40{\mu}M$, signifying that the nitrogen critical point for the growth of the two species stands between 13.5 and $40{\mu}M$. Also, even though both of the two species continued their linear growth with the increase of phosphate ($PO_4^{2-}$) concentrations at less than the $4.05{\mu}M$, there were no any significant differences in growth rates with the increase of phosphate concentrations at more than $4.05{\mu}M$, signifying that the phosphate critical point for the growth of the two species stands between 1.35 and $4.05{\mu}M$. On the other hand, C. polykrikoides has made blooms at the oligotrophic environment near Narodo and Namhaedo where the concentration of DIN and DIP are less than 1.2 and $0.3{\mu}M$, respectively. We attributed this phenomenon to its own ecological characteristics of diel vertical migration through which C. polykrikoides could uptake enough nutrients from the deep sea water near bottom during the night time irrespective of the lower nutrient pools in the surface water.

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Pilot-scale Applications of a Well-type Reactive Barrier using Autotrophic Sulfur-oxidizers for Nitrate Removal (독립영양 황탈질 미생물을 이용한 관정형 반응벽체의 현장적용성 연구)

  • Lee, Byung-Sun;Um, Jae-Yeon;Lee, Kyu-Yeon;Moon, Hee-Sun;Kim, Yang-Bin;Woo, Nam-C.;Lee, Jong-Min;Nam, Kyoung-Phile
    • Journal of Soil and Groundwater Environment
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    • v.14 no.3
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    • pp.40-46
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    • 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.