• Title/Summary/Keyword: 16S rRNA analysis

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The Fermentative Hydrogen Production in Trickling Bed Biofilter Filled with Hydrophilic-and Hydrophobic-Media (소수성 및 친수성 담체를 이용한 Trickling Bed Biofilter의 생물학적 수소생산)

  • Jeon, Byung-Seung;Lee, Sun-Mi;Kim, Yong-Hwan;Gu, Man-Bock;Chae, Hee-Jeong;Sang, Byoung-In
    • Journal of Hydrogen and New Energy
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    • v.17 no.4
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    • pp.379-388
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    • 2006
  • Two mesophilic trickling bed bioreactors filled with two different types of media, hydrophilic- and hydrophobic-cubes, were designed and conducted for hydrogen production under the anaerobic fermentation of sucrose. Each bioreactor consisted of the column packed with polymeric cubes and inoculated with heat-treated sludge obtained from anaerobic digestion tank. A defined medium containing sucrose was fed by the different hydraulic retention time(HRT), and recycle rate. Hydrogen concentrations in gas-phase were constant, averaging 40% of biogas throughout the operation. Hydrogen production rate was increased till $10.5\;L{\cdot}h^{-1}{\cdot}L^{-1}$ of bioreactor when influent sucrose concentrations and recycle rates were varied. At the same time, the hydrogen production rate with hydrophobic media application was higher than its hydrophilic media application. No methane was detected when the reactor was under a normal operation. The major fermentation by-products in the liquid effluent of the both trickling biofilters were acetate, butyrate and lactate. In order to run in the long term operation of both reactor filled with hydrophilic and hydrophobic media, biofilm accumulation on hydrophilic media and biogas produced should be controlled through some process such as periodical backwashing or gas-purging. Four sample were collected from each reactor on the opposite hydrogen production rate, and their bacterial communities were compared by terminal restriction fragment length polymorphism (T-RFLP) analysis of PCR products generated using bacterial 16s rRNA gene primers (8f and 926r). It was expressed a marked difference in bacterial communities of both reactors. The trickling bed bioreactor with hydrophobic media demonstrates the feasibility of the process to produce hydrogen gas. A likely application of this reactor technology can be hydrogen gas recovery from pre-treatment of high carbohydrate-containing wastewaters.

Antifouling Activity towards Mussel by Small-Molecule Compounds from a Strain of Vibrio alginolyticus Bacterium Associated with Sea Anemone Haliplanella sp.

  • Wang, Xiang;Huang, Yanqiu;Sheng, Yanqing;Su, Pei;Qiu, Yan;Ke, Caihuan;Feng, Danqing
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.460-470
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    • 2017
  • Mussels are major fouling organisms causing serious technical and economic problems. In this study, antifouling activity towards mussel was found in three compounds isolated from a marine bacterium associated with the sea anemone Haliplanella sp. This bacterial strain, called PE2, was identified as Vibrio alginolyticus using morphology, biochemical tests, and phylogenetic analysis based on sequences of 16S rRNA and four housekeeping genes (rpoD, gyrB, rctB, and toxR). Three small-molecule compounds (indole, 3-formylindole, and cyclo (Pro-Leu)) were purified from the ethyl acetate extract of V. alginolyticus PE2 using column chromatography techniques. They all significantly inhibited byssal thread production of the green mussel Perna viridis, with $EC_{50}$ values of $24.45{\mu}g/ml$ for indole, $50.07{\mu}g/ml$ for 3-formylindole, and $49.24{\mu}g/ml$ for cyclo (Pro-Leu). Previous research on the antifouling activity of metabolites from marine bacteria towards mussels is scarce. Indole, 3-formylindole and cyclo (Pro-Leu) also exhibited antifouling activity against settlement of the barnacle Balanus albicostatus ($EC_{50}$ values of 8.84, 0.43, and $11.35{\mu}g/ml$, respectively) and the marine bacterium Pseudomonas sp. ($EC_{50}$ values of 42.68, 69.68, and $39.05{\mu}g/ml$, respectively). These results suggested that the three compounds are potentially useful for environmentally friendly mussel control and/or the development of new antifouling additives that are effective against several biofoulers.

Selection of Lactic Acid Bacteria with Antibacterial Activity for Extension of Kimchi Shelf-life (김치의 저장성 향상을 위한 항균활성 우수 유산균 선발)

  • Choi, Hak-Jong;Kim, Yu Jin;Lee, Na Ra;Park, Hae Woong;Jang, Ja Young;Park, Sung-Hee;Kang, Miran;Kim, Hyun Ju;Lee, Jong-Hee;Lee, Jong-Hoon;Pyun, Yu-Ryang;Kim, Tae-Woon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.2
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    • pp.328-332
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    • 2014
  • A survey was conducted on the isolation of lactic acid bacteria with antibacterial activity to extend kimchi shelf-life. Antibacterial activity was tested against bacteria associated with acidification of kimchi, including Lactobacillus plantarum, Pediococcus pentosaceus, and Lactobacillus sakei, using agar-well diffusion assay. Two isolates from kimchi were identified as Lactococcus lactis subsp. lactis and Lactobacillus brevis by 16S rRNA gene sequence analysis and API 50 CHL assay, and they showed antibacterial effects against indicator strains. The isolates displayed acid tolerance at pH 3.5, salt tolerance in 5% NaCl, and growth at $4^{\circ}C$. These result imply that the selected strains might be used to extend kimchi shelf-life as a potential starter.

Comparative analysis of the microbial communities in raw milk produced in different regions of Korea

  • Kim, In Seon;Hur, Yoo Kyung;Kim, Eun Ji;Ahn, Young-Tae;Kim, Jong Geun;Choi, Yun-Jaie;Huh, Chul Sung
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.11
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    • pp.1643-1650
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    • 2017
  • Objective: The control of psychrotrophic bacteria causing milk spoilage and illness due to toxic compounds is an important issue in the dairy industry. In South Korea, Gangwon-do province is one of the coldest terrains in which eighty percent of the area is mountainous regions, and mainly plays an important role in the agriculture and dairy industries. The purposes of this study were to analyze the indigenous microbiota of raw milk in Gangwon-do and accurately investigate a putative microbial group causing deterioration in milk quality. Methods: We collected raw milk from the bulk tank of 18 dairy farms in the Hoengseong and Pyeongchang regions of Gangwon-do. Milk components were analyzed and the number of viable bacteria was confirmed. The V3 and V4 regions of 16S rRNA gene were amplified and sequenced on an Illumina Miseq platform. Sequences were then assigned to operational taxonomic units, followed by the selection of representative sequences using the QIIME software package. Results: The milk samples from Pyeongchang were higher in fat, protein, lactose, total solid, and solid non-fat, and bacterial cell counts were observed only for the Hoengseong samples. The phylum Proteobacteria was detected most frequently in both the Hoengseong and Pyeongchang samples, followed by the phyla Firmicutes and Actinobacteria. Notably, Corynebacterium, Pediococcus, Macrococcus, and Acinetobacter were significantly different from two regions. Conclusion: Although the predominant phylum in raw milk is same, the abundances of major genera in milk samples were different between Hoengseong and Pyeongchang. We assumed that these differences are caused by regional dissimilar farming environments such as soil, forage, and dairy farming equipment so that the quality of milk raw milk from Pyeongchang is higher than that of Hoengseong. These results could provide the crucial information for identifying the microbiota in raw milk of South Korea.

Molecular Epidemiology of Legionella pneumophila Isolated from Water Supply Systems in Seoul, Korea (서울시 수계시설에서 분리된 Legionella pneumophila의 분자역학적 특성)

  • Jeon, Su Jin;Jung, Ji Hun;Seung, Hyun Jung;Kim, Chang Kyu;Jin, Young Hee;Oh, Young Hee;Choi, Sung Min;Chae, Young Zoo
    • Journal of Environmental Health Sciences
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    • v.39 no.2
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    • pp.166-177
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    • 2013
  • Objectives: The genus Legionella is common in aquatic environments. Some species of Legionella are recognized as potential opportunistic pathogens for human, notably Legionella pneumophila that causes, Legionellosis. Thus, we investigated the contamination of Legionella pneumophila on water supply systems in Seoul, including cooling towers, public baths, hospitals and fountains. Methods: The existence of 16S rRNA and mip gene of L. pneumophila was confirmed in the genome of the isolated strains by PCR. Results: During the summer season of 2010 and 2011, Legionella pneumophila were detected from 163 samples (21.1%) out of 772 samples collected. Among the 163 strains of L. pneumophila, eighty one isolates belonged to serogroup 1 (57.4%), 23 isolates were serogroup 5 (16.3%), 21 isolates were serogroup 6 (14.9%), 8 isolates were serogroup 2 (5.79%), and 8 isolates were identified in serogroup 3 (5.7%). Through PFGE (pulsed-field gel electrophoresis) analysis using Sfi I, genetic types of L. pneumophila were classified into five (A to E) patterns by the band similarity with excess of 70% from public baths. Conclusions: The PFGE patterns of the serotypes showed a tendency for diversity of L. pneumophila. Our results suggest the existence of serological and genetic diversity among the L. pneumophila isolates.

Correlation analysis of muscle amino acid deposition and gut microbiota profile of broilers reared at different ambient temperatures

  • Yang, Yuting;Gao, Huan;Li, Xing;Cao, Zhenhui;Li, Meiquan;Liu, Jianping;Qiao, Yingying;Ma, Li;Zhao, Zhiyong;Pan, Hongbin
    • Animal Bioscience
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    • v.34 no.1
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    • pp.93-101
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    • 2021
  • Objective: Temperature could influence protein and amino acid deposition as well as gut microbiota profile and composition. However, the specific effects of ambient temperature on amino acids deposition and gut microbiota composition remain insufficiently understood. Methods: A total of 300 one-day-old Avian broilers were randomly divided into three groups and reared at high, medium, and low temperature (HT, MT, and LT), respectively. Breast muscle and fecal samples were collected for amino acid composition analysis and 16S rRNA gene sequence analysis. Results: Our data showed that compared to the MT group, there was a decrease of muscle leucine and tyrosine (p<0.05), as well as an increase of methionine in the HT group (p<0.05) and a decrease of serine in the LT group. Examination of microbiota shift revealed that at genus level, the relative abundance of Turicibacter and Parabacteroides was increased in the HT group (p<0.05) and that the relative abundances of Pandoraea, Achromobacter, Prevotella, Brevundimonas, and Stenotrophomonas in the LT group were higher than those in the MT group (p<0.05). In addition, there were substantial correlations between microbes and amino acids. In the HT group. Turicibacter was negatively correlated with aspartic acid and tyrosine, whereas Parabacteroides was positively correlated with methionine (p<0.05). In the LT group, there were multiple positive correlations between Achromobacter and arginine, isoleucine or tyrosine; between Prevotella and cysteine or phenylalanine; between Brevundimonas and cysteine; and between Stenotrophomonas and cysteine as well as a negative correlation between Stenotrophomonas and serine. Conclusion: Our findings demonstrated that amino acid content of breast muscle and intestinal microbiota profile was affected by different ambient temperatures. Under heat exposure, augmented abundance of Parabacteroides was correlated with elevated methionine. Low temperature treatment may affect muscle tyrosine content through the regulation of Achromobacter.

Characterization of Photobacterium sp. YW2207 isolated from rainbow trout (Oncorhynchus mykiss) raised in a fresh water farm in South Korea (국내 양식 무지개송어(Oncorhynchus mykiss)에서 분리된 Photobacterium sp. YW2207의 특성)

  • Hyunwoo Kim;Eunsup Lee;Sung Jun Lee;Haneul Kim;So-Ra Han;Tae-Jin Oh;Myoung Sug Kim;Soo-Jin Kim;Se Ryun Kwon
    • Journal of fish pathology
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    • v.36 no.2
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    • pp.251-261
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    • 2023
  • Photobacterium sp. YW2207 was isolated from rainbow trout raised in a fish farm located in Yeongwol-gun, Gangwon Province, South Korea. Based on 16S rRNA sequence analysis and phylogenetic analysis, it was confirmed that Photobacterium sp. YW2207 showed 100% similarity with Photobacterium piscicola and Photobacterium phosphoreum, and 94.6% similarity with P. damselae subsp. damselae. Biochemical analysis revealed that Photobacterium sp. YW2207 is a Gram-negative, motile bacterium with a cell size of 1.5~3×3~5 ㎛. The bacteria were cultured on nutrient agar, brain heart infusion agar, Muller-Hinton agar, tryptic soy agar, and thiosulfate citrate bile sucrose agar with NaCl concentrations ranging from 0 to 2.5%. The API50CHE and API20E tests indicated lower utilization capabilities compared to the P. damselae strains provided in the API database. Furthermore, unlike most Photobacterium species, Photobacterium sp. YW2207 presented negative for catalase test. Results from the flow cytometric measurement indicated that Photobacterium sp. YW2207 exhibited a more diverse distribution of cell sizes and had larger cell sizes compared with P. damselae subsp. damselae. Minimum inhibitory concentration tests showed that Photobacterium sp. YW2207 had low susceptibility to β-Lactam and aminoglycoside antibiotics, while having high susceptibility to tetracycline, doxycycline, and quinolone antibiotics. Pathogenicity on rainbow trout revealed that an immersion of 1×105 CFU/ml did not cause mortality or clinical symptoms.

Antifungal Activity of Bacillus sp. BCNU 2003 against the Human Pathogenic Fungi (인체 병원성 진균에 대한 Bacillus sp. BCNU 2003의 항진균 효과)

  • Choi, Hye-Jung;Yang, Uk-Hee;Kim, Ya-Ell;Choi, Yeon-Hee;Ahn, Cheol-Soo;Jeong, Young-Kee;Kim, Dong-Wan;Joo, Woo-Hong
    • Journal of Life Science
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    • v.20 no.2
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    • pp.269-274
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    • 2010
  • An antifungal antibiotic-producing strain, BCNU 2003, was isolated from forest soil in Korea. The morphological and physiological characters, and 16S rRNA sequences analysis of strain BCNU 2003 identified this strain as Bacillus genus. The Bacillus sp. BCNU 2003 showed strong antifungal activities against Aspergillus niger, Trichophyton mentagrophytes and Trichophyton rubrum with inhibition ranging from 62.05 to 63.49% by using dual culture technique. Bacillus sp. BCNU 2003 produced a maximum level of antifungal substances under aerobic incubation at 28oC and pH 6.5-7.2 for 6 days in LB broth. Ethyl acetate extract of the cultured broth showed strong antifungal activity and a broad antifungal spectrum against various pathogenic fungi. The minimum inhibitory concentration (MIC) values for its active extracts ranged between 0.0625 mg/ml and 1 mg/ml. In addition, Bacillus sp. BCNU 2003 was determined to have the ability to produce enzymes such as amylase, protease, gelatinase and catalase.

Characterization of Bacillus licheniformis SCK A08 with Antagonistic Property Against Bacillus cereus and Degrading Capacity of Biogenic Amines (Bacillus cereus에 대한 길항적 저해 작용과 biogenic amines 분해 능력을 지닌 Bacillus licheniformis SCK A08 균의 특성)

  • Lee, Eon Sil;Kim, Yong Sang;Ryu, Myeong Seon;Jeong, Do Yeon;Uhm, Tai Boong;Cho, Sung Ho
    • Journal of Food Hygiene and Safety
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    • v.29 no.1
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    • pp.40-46
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    • 2014
  • We have screened Bacillus strains suitable for the fermentation of soybean products with respect to the control of Bacillus cereus and the reduction of biogenic amines. Of 26 isolates, a strain named as the SCK A08 carried antimicrobial activity against B. cereus and Staphylococcus aureus, major food poisoning species in soybean products. PCR analysis revealed that the SCK A08 strain did not contain genes for Bacillus cereus toxins including nonhemolytic enterotoxin, hemolytic enterotoxin, cytotoxin K, cereulide and certrax. The SCK A08 strain could degrade histamine, tyramine, putrescine, and cadaverine by 67.41%, 76.59%, 57.32%, and 50.69%, respectively, during fermentation in cooked soybeans containing 0.5% (w/w) of each biogenic amine. The morphological and biochemical properties and phylogenetic relationships based on 16S rRNA gene sequences indicated that the isolate was most closely related to Bacillus licheniformis. Use of the strain SCK A08 would be a potential measure to overcome two hygienic problems that were frequently faced during manufacture of traditionally fermented soybean products.

The effect of antagonists produced by Paenibacillus polymyxa CK-1 on the growth of Trichoderma sp. (Paenibacillus polymyxa CK-1이 생산한 길항물질이 Trichoderma sp. 생육에 미치는 영향)

  • Lee, Sang-Won;Choi, Jin-Sang;Kim, Chul-Ho
    • Journal of Mushroom
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    • v.12 no.3
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    • pp.201-208
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    • 2014
  • The separation of the bacteria inhibiting Trichoderma sp. mold, the strain causing blue mold disease that occurs frequently when cultivating mushroom while carrying out the efficient fermentation of mushroom medium, from the growth was done. In about 200 strains isolated primarily from fungus garden samples, 6 strains were secondly isolated, which had fast growth rates and a clear zone on the plate medium of SM, AM, and CM. Among the 6 strains isolated, the C-1 strain showed high enzymatic activity of cellulase, amylase, and protease, and strong antibacterial activity for the T. virens and T. harzianum, selected finally. The selected C-1 strain was identified as Paenibacillus polymyxaby the result of the identification by Bergey's Manual of Systematic Bacteriology and the analysis of the nucleotide sequence of 16S rRNA, and named as P. polymyxa CK-1. In reviewing the growth conditions of the P. polymyxa CK-1 strain, the optimum cultivation temperature was $45^{\circ}C$, and the optimum pH for growth was in the range of 6.0~7.0. Appropriate incubation time of P. polymyxa CK-1 for the growth inhibition of the fungus T. virens and T. harzianum was 22 to 36 hours. And the fungal growth was not observed, even when leaving two molds inoculated on each petri dishes, which were treated with 24 hour culture solution of P. polymyxa CK-1 strain for 10 days. As a result of studying the thermal stability of the antagonists produced by the P. polymyxa CK-1 strain, no mycelial growth of the two fungi was observed in the test group treated for 20 minutes at $60^{\circ}C$ and $100^{\circ}C$, but mycelial growth was slightly observed in the test group treated for 20 minutes at $121^{\circ}C$. As aresult of reviewing the impact of the P. polymyxa CK-1 culture medium on mushroom mycelial growth, it showed no effect on a variety of mushroom mycelial growth including enoki mushroom and shiitake mushroom.