• Title/Summary/Keyword: 16S-rRNA

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Respiratory Microbiome in Children (소아의 호흡기 미생물군 유전체)

  • Kim, Dong Hyun
    • Pediatric Infection and Vaccine
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    • v.26 no.3
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    • pp.129-139
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    • 2019
  • The human respiratory tract hosts both pathogenic and commensal bacteria. The development of well-conserved 16S rRNA sequencing and culture-independent techniques has enabled many achievements in the study of the human microbiome. Microbial composition of the respiratory tract in early childhood has been shown to correlate to respiratory health in later stages of life. This review highlights current understandings of respiratory microbiota development in healthy children, examples of microbial interactions, impacts on the host immune system, and the relationship between respiratory tract microbiome and respiratory health.

Isolation and Identification of Vibrio Species Contaminated in Imported Frozen Seafoods (수입냉동 어패류에 오염되어 있는 Vibrio속 세균의 분리 및 동정)

  • 윤영준;김도연;이실한;이우윤;고영환;김승곤;김정완
    • Journal of Food Hygiene and Safety
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    • v.15 no.2
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    • pp.128-136
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    • 2000
  • Twenty-four Vibrio strains were isolated from imported frozen seafoods and identified according to their physiological and biochemical properties. They included two V cholerae non-01 sp., two V. diazotrophicus sp., one V. hollisae sp., five V. natriegens sp., eight V. fluvialis sp., and four V. nereis sp.. Two of them were not identified as Vibrio species. When these strains were tested using API-2OE kit fur identification, however, only the results for two V. cholerae and five of the V. fluvialis strains matched the results obtained previously. Due to the importance of detecting V cholerae from foods, phylogenetic identification of the strains was attempted based on restriction fragment length polymorphism (RFLP) of the 16S rDNAs amplified by PCR. The results suggested that the two strains had identical RFLP patterns which were more closely related to that of V. proteolyticus than V. cholerae. The problems associated with identification of pathogens originated from seafoods demand development of accurate and rapid identification methods.

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Molecular Identification of Vaginal Lactobacillus spp. Isolated from Korean Women

  • CHANG, CHUNG EUN;SYLVIA I. PAVLOVA;LIN TAO;EUN-KI KIM;SEUNG CHUL KIM;HYUN SHIK YUN;JAE-SEONG SO
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.312-317
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    • 2002
  • Indigenous lactobacilli were isolated from vaginas of Korean women for possible use in ecological treatment of bacterial vaginosis. Vaginal swab samples were obtained from a gynecological clinic and streaked on Rogosa SL agar plates to select the most predominant lactobacilli in each sample. The preliminary identification of the isolates as lactobacilli was based on microscopic observation of Gram-positive rod-shaped cell morphology. The initial characterization was performed on 108 isolates in terms of their cell surface hydrophobicity (CSH), antimicrobial activity, and hydrogen peroxide (H₂O₂) production capability, and 10 isolates were then selected for further molecular identification. For a rapid procedure to identify lactobacilli, polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analyses of the l6S rRNA genes were applied. The 10 selected lactobacilli and 9 different reference strains of Lactobacillus spp. were characterized by PCR-RFLP where the amplified l6S rDNA was digested with 7 different restriction endonucleases prior to analysis. DNA sequencing of the 16S rRNA gene of one particular isolate, KLB 46, that had been identified as L. crispatus by the PCR-RFLP analysis, further confirmed its identity as L. crispatus.

Distribution and Identification of Halophilic Bacteria in Solar Salts Produced during Entire Manufacturing Process (천일염 생산공정별 미생물 분포 조사 및 호염미생물 동정)

  • Na, Jong-Min;Kang, Min-Seung;Kim, Jin-Hyo;Jin, Yong-Xie;Je, Jeong-Hwan;Kim, Jung-Bong;Cho, Young-Sook;Kim, Jae-Hyun;Kim, So-Young
    • Microbiology and Biotechnology Letters
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    • v.39 no.2
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    • pp.133-139
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    • 2011
  • In this study, we determined the changes in microbial numbers in solar salts according to the manufacturing process and storage duration. The salt samples were harvested from salt farms in Shinan (area 2) and Yeonggwang (area 1). They were serially diluted ten-fold and then placed on 4 kinds of cultivable media (mannitol salt agar, eosin methylene blue, plate count agar, and trypticase soy agar). After incubation, we obtained 62 halophilic isolates from the salt samples. Coliform and general bacteria were not detected in all salt samples. By 16S rRNA sequencing analysis, we found 12 kinds of halophilic bacteria belonging to the genera Halobacillus, Halomonas, Bacillus, Idiomarina, Marinobacter, Pseudoalteromonas, Vibrio, Salinivibrio, Virgibacillus, Alteromonas, Staphylococcus and some un-known stains. In our study, we discovered two novel species that have a 16S rDNA sequence similarity below 97%.

Phylogenetic diversity and UV resistance analysis of radiation-resistant bacteria isolated from the water in Han River (한강물로부터 분리된 방사선 내성 세균들의 계통학적 다양성 및 UV 내성 분석)

  • Lee, Jae-Jin;Joo, Eun Sun;Lee, Do Hee;Jung, Hee-Young;Kim, Myung Kyum
    • Korean Journal of Microbiology
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    • v.52 no.1
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    • pp.65-73
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    • 2016
  • The aim of this study was to investigate the UV-resistance of radiation-resistant bacteria isolated from the water of Han River, South Korea. The water sample was irradiated with 3 kGy gamma radiation prior to isolation. Radiation-resistant bacterial strains were isolated by standard serial dilution method on R2A and 1/10 diluted R2A agar. The resulting purely isolated 60 cultures of bacteria were analysed for UV resistance and used in further studies. Based on the comparative analyses of 16S rRNA gene sequences, the bacterial isolates were divided into 3 phyla (4 genera): the phylum Deinococcus-Thermus (the genus Deinococcus) was 61.7%, Bacteroidetes (Hymenobacter and Spirosoma) was 23.4%, and Firmicutes (Exiguobacterium) was 15%. The results suggested that twenty-nine isolates are candidates new species belonging to Deinococcus, Hymenobacter, and Spirosoma, or other new genera. Nine bacterial strains were selected among the novel candidates and the UV-resistance analysis was conducted. All the candidate bacterial strains showed high UV resistance, similar to that of D. radiodurans R1.

Isolation and characterization of lactic acid bacteria for use as silage additives (사일리지 제조를 위한 유산균 탐색 및 특성연구)

  • Ro, Yu-Mi;Lee, Gwan-Hyeong;Park, InCheol;Kim, Wan-Gyu;Han, Byeong-Hak;You, Jaehong;Ahn, Jae-Hyung
    • Korean Journal of Microbiology
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    • v.52 no.4
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    • pp.444-454
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    • 2016
  • Sixteen lactic acid bacterial strains were isolated from silage and cow dung samples, and characterized to identify their potential as silage additives. They were identified as the members of the genera Lactobacillus, Enterococcus, and Weissella, and clustered into nine groups based on the sequences of the genes for 16S rRNA, RNA polymerase alpha subunit, 60-kDa heat shock protein, and phenylalanyl-tRNA synthase alpha subunit. Among them, the three strains which were genetically similar to L. plantarum showed the fastest growth and pH decrease in MRS and rye extract media, the highest numbers of available carbohydrates, and the widest ranges of pH, temperature, and salinity for growth. In addition, they showed no amplified DNA products in the PCR examination targeting the genes for the production of biogenic amines, and the MRS media where they had been cultured showed relatively high inhibition effect against the growth of silage-spoiling microorganisms, including fungi, yeast, and clostridia. The results suggest that these strains are good candidates for silage additives. However, the rye extract media where the lactic acid bacteria had been cultured had no effect on or stimulated the growth of the silage-spoiling microorganisms, and the causes must be established for the practical use of the lactic acid bacteria as silage additives.

Genomic Analysis of a Freshwater Actinobacterium, "Candidatus Limnosphaera aquatica" Strain IMCC26207, Isolated from Lake Soyang

  • Kim, Suhyun;Kang, Ilnam;Cho, Jang-Cheon
    • Journal of Microbiology and Biotechnology
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    • v.27 no.4
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    • pp.825-833
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    • 2017
  • Strain IMCC26207 was isolated from the surface layer of Lake Soyang in Korea by the dilutionto-extinction culturing method, using a liquid medium prepared with filtered and autoclaved lake water. The strain could neither be maintained in a synthetic medium other than natural freshwater medium nor grown on solid agar plates. Phylogenetic analysis of 16S rRNA gene sequences indicated that strain IMCC26207 formed a distinct lineage in the order Acidimicrobiales of the phylum Actinobacteria. The closest relative among the previously identified bacterial taxa was "Candidatus Microthrix parvicella" with 16S rRNA gene sequence similarity of 91.7%. Here, the draft genome sequence of strain IMCC26207, a freshwater actinobacterium, is reported with the description of the genome properties and annotation summary. The draft genome consisted of 10 contigs with a total size of 3,316,799 bp and an average G+C content of 57.3%. The IMCC26207 genome was predicted to contain 2,975 protein-coding genes and 51 non-coding RNA genes, including 45 tRNA genes. Approximately 76.8% of the protein coding genes could be assigned with a specific function. Annotation of the IMCC26207 genome showed several traits of adaptation to living in oligotrophic freshwater environments, such as phosphorus-limited condition. Comparative genomic analysis revealed that the genome of strain IMCC26207 was distinct from that of "Candidatus Microthrix" strains; therefore, we propose the name "Candidatus Limnosphaera aquatica" for this bacterium.

Isolation and Identification of Lactic Acid Bacteria Inhibiting Gastro-intestinal Pathogenic Bacteria of Domestic Animal. (가축 소화기 병원성 세균을 저해하는 유산균의 분리 및 동정)

  • Lee, Jae-Yeon;Hwang, Kyo-Yeol;Kim, Hyun-Soo;Kim, Geun;Sung, Soo-Il
    • Microbiology and Biotechnology Letters
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    • v.30 no.2
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    • pp.129-134
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    • 2002
  • To isolate probiotic lactic acid bacteria having superior inhibitory activities against animal gastro-intestinal pathogenic bacteria such as Salmonella gallinarum, Staphylococcus aureus and Escherichia coli, 130 strains were initially isolated from the small intestines of Korean native chickens and 7 lactic acid bacteria were finally selected. By using API CHL kit and 16S rRNA sequencing method, the selected lactic acid bacteria were found to be belonged to genus Lactobacillus except BD14 identified as Pediococcus pentosaceus. Especially, Lactobacillus pentosus K34 showed the highest resistancy to both of HCl and bile salt, as well as the highest inhibitory activities against S. gallinarum, S. aureus and E. coli. All the selected strains were sensitive to various antibiotics such as neomycin, erythromycin, cephalosporin, amoxicillin/clavulanic acid, ampicillin, oxytetracycline, but resistant to ciprofloxacin. All the selected strains except BL strain were resistant to colistin and streptomycin, and BD14, BD16, K34 strains were resistant to gentamicin.

Quantitative Analysis of Human- and Cow-Specific 16S rRNA Gene Markers for Assessment of Fecal Pollution in River Waters by Real-Time PCR

  • Jeong, Ju-Yong;Park, Hee-Deung;Lee, Kyong-Hee;Hwang, Jae-Hong;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.245-253
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    • 2010
  • The base sequences representing human- and cow-specific 168 rRNA gene markers identified in a T-RFLP analysis were recovered from clone libraries. The human- and cow-specific primers were designed from these sequences and their specificities were analyzed with fecal DNAs from human, cow, and pig. The AllBac primer set showed positive results for all human, cow, and pig samples, whereas the human-specific primer set showed positive result only for the human sample but not for the cow or pig samples. Likewise, the cow-specific primer set showed positive results only for the cow sample but not for the human or pig samples. Real-time PCR assay with these primers was developed for the identification and quantification of fecal pollution in the river water. The human- and cow-specific markers were detected in the order of 9 $\log_{10}$ copies per gram wet feces, which were two orders of magnitude lower than those of total Bacteroidales. For the river water samples, the human-specific marker was detected in $1.7-6.2\;\log_{10}$ copies/100 ml water, which was 2.4-4.9 orders of magnitude lower than those of total Bacteroidales. There was no significant correlation between total Bacteroidales and conventional fecal indicators, but there was a high correlation between Bacteroidales and the human-specific marker. This assay could reliably identify and quantify the fecal pollution sources, enabling effective measures in the watersheds and facilitating water quality management.

Development of Species-specific PCR Primers for Detecting Peptoniphilus mikwangii

  • Park, Soon-Nang;Lee, Junhyeok;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.42 no.3
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    • pp.143-147
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    • 2017
  • In a previous study, Peptoniphilus mikwangii was isolated from the human oral cavity as a new species. The purpose of this study was to develop P. mikwangii-specific PCR primers. The PCR primers were designed, based on the nucleotide sequence of 16S ribosomal RNA (16S rDNA). The specificity of the primers was tested using genomic DNAs of 3 strains of P. mikwangii and 27 strains (27 species) of non-P. mikwangii bacteria. The sensitivity of primers sensitivity was determined using PCR, with serial dilutions of the purified genomic DNAs (4 ng to 4 fg) of P. mikwangii KCOM $1628^T$. The data showed that P. mikwangii-specific qPCR primers (B134-F11/B134-R1 & B134-F5/B134-R5) could detect only P. mikwangii strains, and 400 fg or 40 fg of P. mikwangii genome DNA. These results suggest that PCR primers are useful in detecting P. mikwangii from the oral cavity.