• Title/Summary/Keyword: rDNA sequencing

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Phylogenetic Analysis of Oligotrophic Bacteria Found in Potable Groundwater (음용 지하수중에 분포하는 저영양세균의 계통학적 해석)

  • ;Tomoyoshi Hashimoto
    • Korean Journal of Microbiology
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    • v.38 no.4
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    • pp.293-298
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    • 2002
  • In order to investigate the ecological aspect of bacteria on groundwater, water samples were collected from various regions. Total of 318 strains were isolated from diluted nutrient broth (DNB) agar medium, and investigated their growth pattern on nutrient broth (NB) medium. As a result, all the isolated strains were divided into two groups, NB and DNB organisms. Growth of DNB organisms were suppressed in full strength NB medium but not in DNB medium, which were called oligotrophic bacteria in this study. Proportion of DNB organisms occurred in the frequency of 50-98% in potable groundwaters (CW, CJ, DPG, CJG1), however, it was 23,46% in polluted site (TJ, NPG1). One hundred and two strains were identified as oligotrophic bacteria and their phylogenetic characteristics were determined by using 16S rDNA sequencing. Based on the phylogenetic analysis, they were found to fall into three major phylogenetic groups: belonging to the Proteobacteria $\alpha$-(49 strains), $\beta$-(50 strains), $\gamma$ -(3 strains) subdivisions. The phylogenetic analysis suggested that microbial diversity of potable groundwater is more complex than that obtained in the past investigation.

Microbial Community Profiling in cis- and trans-Dichloroethene Enrichment Systems Using Denaturing Gradient Gel Electrophoresis

  • Olaniran, Ademola O.;Stafford, William H.L.;Cowan, Don A.;Pillay, Dorsamy;Pillay, Balakrishna
    • Journal of Microbiology and Biotechnology
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    • v.17 no.4
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    • pp.560-570
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    • 2007
  • The effective and accurate assessment of the total microbial community diversity is one of the primary challenges in modem microbial ecology, especially for the detection and characterization of unculturable populations and populations with a low abundance. Accordingly, this study was undertaken to investigate the diversity of the microbial community during the biodegradation of cis- and trans-dichloroethenes in soil and wastewater enrichment cultures. Community profiling using PCR targeting the l6S rRNA gene and denaturing gradient gel electrophoresis (PCR-DGGE) revealed an alteration in the bacterial community profiles with time. Exposure to cis- and trans-dichloroethenes led to the disappearance of certain genospecies that were initially observed in the untreated samples. A cluster analysis of the bacterial DGGE community profiles at various sampling times during the degradation process indicated that the community profile became stable after day 10 of the enrichment. DNA sequencing and phylogenetic analysis of selected DGGE bands revealed that the genera Acinetobacter, Pseudomonas, Bacillus, Comamonas, and Arthrobacter, plus several other important uncultured bacterial phylotypes, dominated the enrichment cultures. Thus, the identified dominant phylotypes may play an important role in the degradation of cis- and trans-dichloroethenes.

Characterization of a bacterium isolated from diseased rock sea bream, Oplegnathus fasciatus, during low water temperature season in Jeju island (저수온기의 제주도 양식 돌돔, Oplegnathus fasciatus, 병어로부터 분리한 병원세균의 특성)

  • Go, Dae-Hui;Jin, Chang-Nam;Lee, Chang-Hun;Park, Geun-Tae;Heo, Mun-Su
    • Journal of fish pathology
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    • v.17 no.2
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    • pp.83-90
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    • 2004
  • The outbreak of haemorrhagic specticemia and lesions in dermis, frequently associated with keratitis have occured on rock sea bream (Oplegnathus fasciatus) reared in Jeju island. A bacterium isolated from the diseased fish identified as Pseudomonas sp. by morphologic, cultural, and biochemical tests. The bacterium was identified Pseudomonas anguilliseptica according to 16S rDNA sequence analysis. The highest growth rate of this strain was obtained at 20 to $\25^circ{C}$ although the disease occurred mostly below $16^\circ{C}$. The challenge test by ip injection revealed that this bacterium was pathogenic to rock sea bream. All experimental fish in challenged by ip injection died within 21 days but there was no death in control group. The bacterium was sensitive to Doxycycline, Chloramphenicol, Ciprofloxacin, Erythromycin, Flumequine, Nalidixic acid, Norfloxacin, and Ofloxacin.

Biochemical and Molecular Identification of Antibacterial Lactic Acid Bacteria Isolated from Kimchi (김치에서 항균활성 유산균의 분리 및 동정)

  • Kim, Soo-Young;Kim, Jong-Doo;Son, Ji-Soo;Lee, Si-Kyung;Park, Kab-Joo;Park, Myeong-Soo
    • Korean Journal of Food Science and Technology
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    • v.43 no.4
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    • pp.446-452
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    • 2011
  • Total 480 lactic acid-producing bacteria were isolated from five kinds of kimchi, and their antibacterial activity was tested against Salmonella enterica serovar Typhimurium, Bacillus subtilis, and Pseudomonas aeruginosa using an agar diffusion assay. Among them, 340 isolates showed a halo on MRS agar against one or more indicator strains, which were identified using multiplex PCR, an API 50CHL kit, and a 16S rDNA sequence analysis. As a result, 169 Lactobacillus plantarum, 20 Lactobacillus fermentum, two Lactobacillus paracasei ssp. paracasei, two Lactobacillus sp., and 15 Pediococcus sp. were identified. This may be the first report on the isolation of antibacterial Lactobacillus fermentum from kimchi.

Sphingobacterium sp. SW-09 Effectively Degrades Phenanthrene, a Polycyclic Aromatic Hydrocarbon, in a Soil Microcosm (Sphingobacterium sp. SW-09에 의한 토양환경에서의 다환 방향족탄화수소인 페난스렌의 분해)

  • Son, Seung-Woo;Chang, Hey-Won;Kim, Sung-Kuk;Chang, Jong-Soo
    • Journal of Life Science
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    • v.21 no.11
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    • pp.1511-1517
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    • 2011
  • We isolated a potent phenanthrene-degrading bacterium from oil-contaminated soils of Suzhou, China, and assessed the potential use of these bacteria for bioremediation of soils contaminated by polycyclic aromatic hydrocarbons (PAHs) in a microcosm. Based on 16S rDNA sequencing, we identified this bacteria as Sphigobacterium sp. SW-09. By PCR amplification, we also identified catechol 2,3-dioxygenase genes (nahH genes) mediating PAH degradation. Staphylococcus sp. KW-07, which has been identified in our previous study, showed potential for use in bioremediation of oil-contaminated soils. In this experiment, we compared the rate of phenanthrene-degradation between Staphylococcus sp. KW-07 and Sphingobacterium sp. SW-09 in a microcosm condition. Newly isolated Sphingobacterium sp. SW-09 showed a higher phenanthrene-degradation rate than that of Staphylococcus sp. KW-07 in soil microcosms. Together, our results suggest that the Sphingobacterim sp. SW-09 strain isolated from the Suzhou area may also be useful in bioremediation of PAH-contaminated soils.

Isolation and Characterization of a Marine Derived Bacterium Glaciecola sp. SL-12 Producing β-agarase (한천분해효소를 생산하는 해양유래 세균 Glaciecola sp. SL-12의 분리 및 특성)

  • Lee, Dong-Geun;Lee, Ok-Hee;Jang, Hyo-Jung;Jang, Min-Kyung;Yoo, Ki-Hwan;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.18 no.1
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    • pp.58-62
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    • 2008
  • A novel agar-degrading bacterium SL-12 was isolated from seashore of Kijang at Busan, Korea, and cultured in marine broth 2216 media. Isolated bacterium SL-12 was identified as Glaciecola genus by 16S rDNA sequencing with 98% identity. The optimum pH of the enzyme activity was 7.0 and the optimum temperature for the reaction was $30^{\circ}C$. The enzyme hydrolyzed neoagarohexaose to yield neoagarobiose as the main product, indicating that the enzyme is ${\beta}$-agarase. Thus, isolated bacterium and the enzyme would be useful for the industrial production of neoagarobiose.

Isolation and characterization of a marine bacterium Thalassomonas sp. SL-5 producing β-agarase (한천분해효소를 생산하는 해양세균 Thalassomonas sp. SL-5의 분리 및 특성)

  • Lee, Dong-Geun;Kim, Nam-Young;Jang, Min-Kyung;Lee, Ok-Hee;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.17 no.1 s.81
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    • pp.70-75
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    • 2007
  • A novel agar-degrading bacterium SL-5 was isolated from seashore of Homigot at Kyung-Buk province, and cultured in marine broth 2216 media. The bacterium SL-5 was identified as Thalassomonas genus by 16S rDNA sequencing with 96% identity. Growth rate was faster at $27^{\circ}C$ than at $37^{\circ}C$ and agarase was produced as growth-related. The optimum pH of the enzyme activity was 7.0 and the optimum temperature for the reaction was $40^{\circ}C$. Although the enzyme had no thermostability, the enzyme activity was remained over 80% at $60^{\circ}C$. The enzyme hydrolyzed neoagarohexaose to yield neoagarobiose as the main product, indicating that the enzyme is $\beta-agarase$. Thus, the enzyme would be useful for the industrial production of neoagarobiose.

Freeze-drying feces reduces illumina-derived artefacts on 16S rRNA-based microbial community analysis (Illumina를 이용한16S rRNA 기반 미생물생태분석에서 분변의 동결건조에 의한 인공적인 시퀀스 생성 감소효과)

  • Kim, Jungman;Unno, Tatsuya
    • Journal of Applied Biological Chemistry
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    • v.59 no.4
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    • pp.299-304
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    • 2016
  • When used for amplicon sequencing, Illumina platforms produce more than hundreds of sequence artefacts, which affects operational taxonomic units based analyses such as differential abundance and network analyses. Nevertheless it has become a major tool for fecal microbial community analysis. In addition, results from sequence-based fecal microbial community analysis vary depending on conditions of samples (i.e., freshness, time of storage and quantity). We investigated if freeze-drying samples could improve quality of sequence data. Our results showed reduced number of possible artefacts while maintaining overall microbial community structure. Therefore, freeze-drying feces prior to DNA extraction is recommended for Illumina-based microbial community analysis.

Effect of Temperature on Growth Rate and Protease Activity of Antarctic Microorganisms (극지 미생물들의 배양온도에 따른 성장률 및 protease activity 영향 연구)

  • Kim, Hyun-Do;Choi, Jong-Il
    • Microbiology and Biotechnology Letters
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    • v.42 no.3
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    • pp.293-296
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    • 2014
  • This study was conducted to investigate the effect of culture temperature on the growth rate and protease activity of Antarctic microorganisms. The Antarctic microorganisms PAMC 25641, 25614, 25719 and 25617 were obtained from the Polar and Alpine Microbial Collection (PAMC) at the Korea Polar Research Institute. These microorganisms were confirmed for the excretion of protease on a plate with skim milk. The identification of microorganisms was carried out using the 16S rDNA sequencing method. PAMC 25641 showed the highest protease activity among the subjects tested, and PAMC 25617 exhibited the highest growth rate. The growth rates of the microorganisms were not affected by temperature, except for PAMC 25617. However, protease activities were increased for all strains in a temperature dependent fashion. These results suggest the possible application of Antarctic microorganisms for the efficient production of low temperature proteases.

Screening of Bifidobacteria for the Development of Probiotics Inhibiting Intestinal Pathogenic Bacteria (장내 유해세균을 억제하는 양돈용 프로바이오틱스 개발을 위한 비피도박테리아 탐색)

  • Lee, Jaeyeon;Shin, Yungoh;Kim, Keun
    • Microbiology and Biotechnology Letters
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    • v.42 no.3
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    • pp.211-218
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    • 2014
  • In order to isolate probiotic lactic acid bacteria possessing high inhibitory activities against porcine and zoonotic pathogens, such as enterotoxigenic E. coli, Salmonella Typhimurium, and Clostridium perfringens, a total of 65 anaerobic strains were initially isolated from a variety of sources including cattle rumen fluids, chicken intestines and swine feces. Four Bifidobacterium strains were selected for their high anti-pathogenic bacterial activities. By using the 16S rDNA sequencing method, three B. boum strains and one B. thermophilum were identified. B. thermophilum demonstrated the best adhesive ability to epithelial cells of swine intestine among the isolates. Indeed, B. thermophilum was seen to have superior characteristics as a probiotic for swine, as judged by their high growth inhibitory activities against various pathogens, and high acid- and bile-tolerance.