• Title/Summary/Keyword: 16S rRNA bacterial identification

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A Method for Comparing Multiple Bacterial Community Structures from 16S rDNA Clone Library Sequences

  • Hur, Inae;Chun, Jongsik
    • Journal of Microbiology
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    • v.42 no.1
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    • pp.9-13
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    • 2004
  • Culture-independent approaches, based on 16S rDNA sequences, are extensively used in modern microbial ecology. Sequencing of the clone library generated from environmental DNA has advantages over fingerprint-based methods, such as denaturing gradient gel electrophoresis, as it provides precise identification and quantification of the phylotypes present in samples. However, to date, no method exists for comparing multiple bacterial community structures using clone library sequences. In this study, an automated method to achieve this has been developed, by applying pair wise alignment, hierarchical clustering and principle component analysis. The method has been demonstrated to be successful in comparing samples from various environments. The program, named CommCluster, was written in JAVA, and is now freely available, at http://chunlab.snu.ac.kr/commcluster/.

Comparison of Airborne Bacterial Communities from a Hog Farm and Spray Field

  • Arfken, Ann M.;Song, Bongkeun;Sung, Jung-Suk
    • Journal of Microbiology and Biotechnology
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    • v.25 no.5
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    • pp.709-717
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    • 2015
  • Airborne bacteria from hog farms may have detrimental impacts on human health, particularly in terms of antibiotic resistance and pathogen zoonosis. Despite human health risks, very little is known about the composition and diversity of airborne bacteria from hog farms and hog-related spray fields. We used pyrosequencing analysis of 16S rRNA genes to compare airborne bacterial communities in a North Carolina hog farm and lagoon spray field. In addition, we isolated and identified antibiotic-resistant bacteria from both air samples. Based on 16S rRNA gene pyrosequence analysis, Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria were the dominant phyla in airborne bacterial communities from both hog farm and spray field sites. Within the Firmicutes genera, Clostridium spp. were more abundant in the hog farm, whereas Staphylococcus spp. were higher in the spray field. The presence of opportunitic pathogens, including several Staphylococcus species and Propionibacterium acnes, was detected in both bioaerosol communities based on phylogenetic analysis. The isolation and identification of antibiotic-resistant bacteria from air samples also showed similar results with dominance of Actinobacteria and Proteobacteria in both hog farm and spray field air. Thus, the existence of opportunistic pathogens and antibiotic resistant bacteria in airborne communities evidences potential health risks to farmers and other residents from swine bioaerosol exposure.

Description of 17 unrecorded bacterial species isolated from freshwater showing antibiotic resistance in Korea

  • Baek, Kiwoon;Kim, Eui-Jin;Han, Ji-Hye;Choi, Ahyoung
    • Korean Journal of Environmental Biology
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    • v.38 no.2
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    • pp.289-298
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    • 2020
  • As part of the research program "2018 Rapid screening and identification of freshwater microorganisms using MALDI-TOF/MS library" freshwater samples were collected from a branch of the Nakdong River. Almost 300 antibiotic-resistant bacterial strains were isolated from freshwater samples and subsequently identified by 16S rRNA gene sequencing. Seventeen strains among the isolates shared high 16S rRNA gene sequence similarity (>99.0%) with known species that were not previously recorded in Korea, and each of the isolates also formed a robust phylogenetic clade with the closest species. These species were phylogenetically diverse, belonging to four phyla, seven classes, 10 orders, and 13 genera. At the genus and class level, the previously unrecorded species belonged to Rhodovarius, Xanthobacter, and Shinella of the class Alphaproteobacteria; Ottowia, Simplicispira, and Zoogloea of Betaproteobacteria; Pseudomonas, Acinetobacter, and Shewanella of Gammaproteobacteria; Arcobacter of Epsilonproteobacteria; Sphingobacterium of Sphingobacteriia; Trichococcus of Bacilli; and Leucobacter of Actinobacteria. The previously unrecorded species were further characterized by examining their gram-staining, colony and cell morphology, biochemical properties, and phylogenetic position.

Molecular identification of coagulase-negative staphylococci by rpoB sequence typing (rpoB 염기서열 분석을 이용한 응고효소 음성 포도알세균 분자 동정)

  • Seong, Won-Jin;Kim, Danil;Kim, Eun-Kyung;Ko, Dae-Sung;Ro, Younghye;Kim, Jae-Hong;Kwon, Hyuk-Joon
    • Korean Journal of Veterinary Research
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    • v.58 no.1
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    • pp.51-55
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    • 2018
  • Bovine mastitis (BM) has resulted in enormous economic loss in the dairy industry and coagulase-negative staphylococci (CNS) have caused subclinical BM. Although VITEK 2 GP ID card (VITEK 2) has been used for CNS identification, the probability of identification varies. The rpoB sequence typing (RSTing) method has been used for molecular diagnosis and epidemiology of bacterial infections. In this study, we undertook RSTing of CNS and compared the results with those of VITEK2 and 16S rRNA gene sequencing. As compared VITEK2, the molecular-based methods were more reliable for species identification; moreover, RSTing provided more molecular epidemiological information than that from 16S rRNA gene sequencing.

Identification and genetic characterization of bacterial isolates causing brown blotch on cultivated mushrooms in Korea

  • 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.37-47
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    • 2024
  • Fluorescent bacteria were isolated from sporocarps that browned into various mushrooms during survey at places of the production in Korea. We examined the pathogenicity, biodiversity, and genetic characteristics of the 19 strains identified as Pseudomonas tolaasii by sequence analysis of 16S rRNA and White Line Assay. The results emphasize the importance of rpoB gene system, fatty acid profiles, specific and sensitive PCR assays, and lipopeptide detection for the identification of P. tolaasii. As a result of these various analyses, 17 strains (CHM03~CHM19) were identified as P. tolaasii. The phylogenetic analysis based on the 16S rRNA gene showed that all strains were clustered closest to P. tolaasii lineage, two strains (CHM01, CHM02) were not identified as P. tolaasii and have completely different genetic characteristics as a result of fatty acids profile, specific and sensitive PCR, lipopetide detection, rpoB sequence and REP-PCR analysis. Pathogenicity tests showed 17 strains produce severe brown discolouration symptoms to button mushrooms and watersoaking of sporophore tissue within three days after inoculation. But two strains did not produce discolouration symptoms. Therefore, these two strains will be further investigated for correct species identification by different biological and molecular characteristics.

Pathogenic bacteria causing rot in commercial soybean sprout cultivation

  • Yun, Sung-Chul;Kim, Yong-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.48 no.2
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    • pp.113-119
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    • 2003
  • Soybean sprout pathogenic bacteria were isolated from the large, deep containers of a commercial factory. Over a period of one year, 40 pathogenic-like bacteria were isolated among a total of 732 isolates. In addition to bacteria previously reported to be associated with rotting, such as Pseudomonas putida and Erwinia carotovora, several other genera were also identified: Acinetobacter spp., Chryseobacterium spp., Klebsiella sp., Pantoea agglomerans, Bacillus sp. Fatty acid methyl ester (FAME) analysis using the Microbial ID (MIDI) system, and 16s rRNA sequence analysis, yielded identical results, confirming the identities of these microorganisms. Several types of selective media were not good for identification and determination of population structure in commercial environments, as colony type was not specific to the genus. There was no dominant bacterium, and we were not able to find the main bacterium responsible for soybean spout rot. Even though we did not identify a major target for controlling rot or screening for resistant cultivars, the results of this study indicated that bacterial rot of soybean sprout is endemic. In addition, it emerged that factory epidemics in summer are not caused by the bacteria isolated in this study.

Characterization of Denitrifying and Dissimilatory Nitrate Reduction to Ammonium Bacteria Isolated from Mud Crab Culture Environment

  • Hastuti, Yuni Puji;Rusmana, Iman;Nirmala, Kukuh;Affandi, Ridwan;Fatma, Yuli Siti
    • Microbiology and Biotechnology Letters
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    • v.49 no.3
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    • pp.432-439
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    • 2021
  • Microbial community plays important roles in the culture environment of mud crab Scylla serrata. One of the environmental management efforts for the cultivation of S.serrata is by stabilizing microorganisms involved in nitrogen cycle process. The availability of dissolved inorganic nitrogen in its culture environment under a recirculating system closely relates to the nitrogen cycle, which involves both anaerobic and aerobic bacterial activities. Anaerobically, there are two major nitrogen compound degradation processes, i.e., denitrification and dissimilatory nitrate reduction to ammonium (DNRA). This study aimed to identify denitrifying and DNRA bacteria isolated from the recirculating cultivation of S. serrata. The water samples were collected from anaerobic filters called close filter system, which is anaerobically conditioned with the addition of varying physical filter materials in the recirculating mud crab cultures. The results showed that three denitrifying bacterial isolates and seven DNRA bacterial isolates were successfully identified. The phylogenetic analysis based on 16S rRNA gene of the denitrifying bacteria revealed that HIB_7a had the closest similarity to Stenotrophomonas daejeonensis strain MJ03. Meanwhile, DNRA bacterial isolate of HIB_92 showed a 100% similarity to Bacillus sonorensis strain N3, Bacillus vallismortis strain VITS-17, Bacillus tequlensis strain TY5, Geobacillus sp. strain DB24, Bacillus subtilis strain A1, and Bacillus mojavensis strain SSRAI21. This study provides basic information denitrifying and DNRA bacterial isolates identity which might have the potential to be applied as probiotics in aquaculture systems in order to maintain optimal environmental conditions.

Characterization of Two Urease-Producing and Calcifying Bacillus spp. Isolated from Cement

  • Achal, Varenyam;Mukherjee, Abhijit;Reddy, M. Sudhakara
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1571-1576
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    • 2010
  • Two bacterial strains designated as CT2 and CT5 were isolated from highly alkaline cement samples using the enrichment culture technique. On the basis of various physiological tests and 16S rRNA sequence analysis, the bacteria were identified as Bacillus species. The urease production was 575.87 U/ml and 670.71 U/ml for CT2 and CT5, respectively. Calcite constituted 27.6% and 31% of the total weight of sand samples plugged by CT2 and CT5, respectively. Scanning electron micrography analysis revealed the direct involvement of these isolates in calcite precipitation. This is the first report of the isolation and identification of Bacillus species from cement. Based on the ability of these bacteria to tolerate the extreme environment of cement, they have potential to be used in remediating the cracks and fissures in various building or concrete structures.

Identification of 12 radiation-resistant bacterial species in the phylum Proteobacteria new to Korea

  • Han, Joo Hyun;Maeng, Soohyun;Park, Yuna;Lee, Sang Eun;Lee, Byoung-Hee;Lee, Ki-eun;Kim, Myung Kyum
    • Journal of Species Research
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    • v.9 no.2
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    • pp.85-104
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    • 2020
  • In 2019, after a comprehensive investigation of indigenous prokaryotic species in Korea, a total of 12 bacterial strains assigned to the phylum Proteobacteria were isolated from soil. With the high 16S rRNA gene sequence similarity (>98.8%) and formation of a robust phylogenetic clade with the closest species, it was determined that each strain belonged to independent, predefined bacterial species. This study identified two species in the family Burkholderiaceae, one species in the family Comamonadaceae, two species in the family Oxalobacteraceae, one species in the family Micrococcaceae, one species in the family Bradyrhizobiaceae, one species in the family Methylobacteriaceae, one species in the family Rhizobiaceae, one species in the family Rhodocyclaceae, and one species in the family Sphingomonadaceae. There is no official report about these 12 species in Korea, so are described as unreported bacterial species in Korea in this study. Gram reaction, basic biochemical characteristics, colony, and cell morphology are also described in the species description section.

Identification of a Cellulase Producing Marine Bacillus sp. GC-1 and GC-4 Isolated from Coastal Seawater of Jeju Island (제주 연안의 해수로부터 분리한 Cellulase 생산균 Bacillus sp. GC-1과 GC-4의 동정)

  • Chi, Won-Jae;Park, Da-Yeon;Temuujin, Uyangaa;Lee, Jong-Yeol;Chang, Yong-Keun;Hong, Soon-Kwang
    • Microbiology and Biotechnology Letters
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    • v.39 no.2
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    • pp.97-103
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    • 2011
  • Two Gram positive bacterial strains, designated strain GC-1 and GC-4, were isolated from coastal seawater near Jeju Island in the Republic of Korea. The two strains were identified as members of the genus Bacillus, based on 16S rRNA gene sequencing and data for physiological characteristics analyses. A subtle difference in physiological and genotypical characteristics has led us to designate the strains GC-1 and GC-4. The strain GC-1 showed a 99.91% similarity in 16S rRNA gene sequencing with B. tequiliensis and B. subtilis subsp. inaquosorum and the strain GC-4 showed a 100% similarity in 16S rRNA gene sequencing with those of B. altitudinis, B. stratosphericus, and B. aerophilus. However, both strains exhibited different physiological and genotypical characteristics in many aspects from those of their phylogenetically closest neighbors listed above, which implies that genus Bacillus has diversified into various species during its evolutionary process.