• Title/Summary/Keyword: 16s rDNA

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Detection and genetic characterization of Lawsonia intracellularis from swine in Korea

  • Chu, Jia-Qi;Hu, Xu-Min;Kim, Myung-Cheol;Park, Chang-Sik;Jun, Moo-Hyung
    • Korean Journal of Veterinary Service
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    • v.33 no.3
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    • pp.223-231
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    • 2010
  • A total of 191 samples collected from the commercial swine farms located in Chungnam province were investigated by PCR to estimate the prevalence of Lawsonia (L.) intracellularis infection. In the group of the pigs with proliferative enteritis, 14 (93.3%) of 15 intestinal samples and 12 (80.0%) of 15 feces were positive in PCR. In contrast, a relatively low positive rate (18.0%, 29 of 161 samples) was determined in the group of normal healthy pigs. The group of pigs over 120 days showed the highest positive rates (26.8%, 15 of 56 samples). In the comparison of the sequences of 210bp for species specific fragments and 301bp for outer membrane protein, the isolates (L1. L2) showed almost 100% identity with the reference L. intracellularis (L08049, USA). For the sequences of partial 16s rDNA, the homologies among the 5 isolates (L1-L5) were 97.4% to 99.3%, and those of 5 sequences (L1-L5) versus 5 overseas reference strains of L. intracellularis ranged from 98.6% to 99.8%. In the comparison of the nucleotide sequences among 5 isolates and other species in Desulfovibrionales showed 82.4 to 99.5% identities. The 5 isolates shared relatively low identities (76.9% to 84.4%) with the species of alpha-proteobacteria. In phylogenetic analysis based on the 16s rDNA sequences, all of the 5 isolates (L1-L5) were located in the same branch with the strains of L. intracellularis that were previously isolated from the pigs in USA and China. Seven strains of Desulfovibrio sp. were clustered in the neighboring branches, whereas alpha and gamma Proteobacteria showed distant relationship with L. intracellularis strains. The present findings suggest that L. intracellularis infection is endemic in the swine farms in the regions, and that the domestic isolates maintained very limited genetic variation.

Taxonomic Characteristics of Nitrogen-Fixing Oligotrophic Bacteria from Forest Soil (산림토양으로부터 분리한 저영양성-질소고정세균의 분류학적 특성)

  • 황경숙
    • Korean Journal of Microbiology
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    • v.37 no.2
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    • pp.114-119
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    • 2001
  • Many isolates from different forest soil layers did not show appreciable growth on full strength of the conventional nutrient broth (NB medium) but grow on its 100-fold dilution (DNB medium). These isolates were divided into four types according to organic nutrient concentration in the growth medium from $1^{-1}\;to\;10^{-4}$dilution of normal NB medium. Oligotrophic bacteria were type II and type IV which grew in $10^{-4}$ dilution of NB (1 mg C/l) medium. Sixty strains were isolated for obligate oligotrophic bacteria. Chemotaxonomic and phylogenetic characteristics of eleven isolates of acetylene-reducing (nitrogen-fixing) oligotrophic bacteria from forest soil were investigated. They showed similar characteristics: the cellular fatty acid mainly consisted of straight-chain unsaturated $C_{18:1}$ (60-84% of total fatty acids). Ubiquinone Q-10 and a high guanine plus-cytosine content(61-64 mol%) were found. Eleven isolates of nitrogen-fixing oligotrophic bacteria were found to be closely related by full 16S rDNA sequence simility and many common taxonomic traits. Analysis of full 16S rDNA sequences of eleven isolates indicated that they were more closely related to Bradyrhizobium (similarity values: 98.1-98.8%), Agromonas, Nitrobacter, and Afipia.

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Analysis of Microbial Community During the Anaerobic Dechlorination of Perchloroethylene and Trichloroethylene (Perchloroethylene과 Trichloroethylene의 혐기적 탈염소화 및 미생물 군집 분석)

  • Lee Jae-Won;Kim Byung-Hyuk;Ahn Chi-Yong;Kim Hee-Sik;Yoon Byung-Dae;Oh Hee-Mock
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.281-286
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    • 2005
  • In this study, the anaerobic enrichment cultivation was performed with the sediments and the dredged soils from the cities of Ulsan, Masan, Yeosu, Gwangyang, Ansan and Seongnam. Acetate as an electron donor and PCE (perchloroethylene) or TCE (trichloroethylene) as an electron acceptor were injected into the serum bottle with an anaerobic medium. After the incubation of 12 weeks, the removal efficiency of PCE was highest at $70\%$ in the treatment with the sediment of Ulsan. Also, the bacterial community structure was analyzed by D-DGGE (double denatured gradient gel electrophoresis) through PCR-based 16S rDNA approaches. The dominant species id the anaerobic enrichment were found to belong to the genus of Desulfovibrio.

Lysinabacillus fusiformis and Paenibacillus alvei Obtained from the Internal of NasutitermesTermites Revealed Their Ability as Antagonist of Plant Pathogenic Fungi

  • Fitriana, Yuyun;Tampubolon, Desi Apriani Teresa;Suharjo, Radix;Lestari, Puji;Swibawa, I Gede
    • The Plant Pathology Journal
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    • v.38 no.5
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    • pp.449-460
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    • 2022
  • This study was performed to reveal phenotypic characters and identity of symbiont bacteria of Nasutitermes as well as investigate their potential as antagonist of plant pathogenic fungi. Isolation of the symbiont bacteria was carried out from inside the heads and the bodies of soldier and worker termite which were collected from 3 locations of nests. Identification was performed using phenotypic test and sequence of 16S ribosomal DNA (16S rDNA). Antagonistic capability was investigated in the laboratory against 3 phytopathogenic fungi i.e., Phytophthora capsici, Ganoderma boninense, and Rigidoporus microporus. Totally, 39 bacterial isolates were obtained from inside the heads and the bodies of Nasutitermes. All the isolates showed capability to inhibit growth of P. capsici, however, 34 isolates showed capability to inhibit growth of G. boninense and 32 isolates showed capability to inhibit growth of R. microporus. Two bacterial strains (IK3.1P and 1B1.2P) which showed the highest percentage of inhibition were further identified based on their sequence of 16S rDNA. The result showed that 1K3.1P strain was placed in the group of type strain and reference strains of Lysinibacillus fusiformis meanwhile 1B1.2P strain was grouped within type strain and reference strains Paenibacillus alvei. The result of this study supply valuable information on the role of symbiont bacteria of Nasutitermes, which may support the development of the control method of the three above-mentioned phytopathogenic fungi.

Genomic DNA Extracted from Ancient Antarctic Glacier Ice for Molecular Analyses on the Indigenous Microbial Communities

  • Lee, Sang-Hoon;Bidle, Kay;Falkowski, Paul;Marchant, David
    • Ocean and Polar Research
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    • v.27 no.2
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    • pp.205-214
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    • 2005
  • From ancient Antarctic glacier ice, we extracted total genomic DNA that was suitable for prokaryotic 16S rDNA gene cloning and sequencing, and bacterial artificial chromosome (BAC) library and end-sequencing. The ice samples were from the Dry Valley region. Age dating by $^{40}Ar/^{39}Ar$ analysis on the volcanic ashes deposited in situ indicated the ice samples are minimum 100,000-300,000 yr (sample DLE) and 8 million years (sample EME) old. Further assay proved the ice survived freeze-thaw cycles or other re-working processes. EME, which was from a small lobe of the basal Taylor glacier, is the oldest known ice on Earth. Microorganisms, preserved frozen in glacier ice and isolated from the rest of the world over a geological time scale, can provide valuable data or insight for the diversity, distribution, survival strategy, and evolutionary relationships to the extant relatives. From the 16S gene cloning study, we detected no PCR amplicons with Archaea-specific primers, however we found many phylotypes belonging to Bacteria divisions, such as Actinobacteria, Acidobacteria, Proteobacteria $({\alpha},\;{\beta},\;and\;{\gamma})$, Firmicutes, and Cytophaga-Flavobacterium-Bacteroid$. BAC cloning and sequencing revealed protein codings highly identical to phenylacetic acid degradation protein paaA, chromosome segregation ATPases, or cold shock protein B of present day bacteria. Throughput sequencing of the BAC clones is underway. Viable and culturable cells were recovered from the DLE sample, and characterized by their 16S rDNA sequences. Further investigation on the survivorship and functional genes from the past should help unveil the evolution of life on Earth, or elsewhere, if any.

Study on Rumen Cellulolytic Bacterial Attachment and Fermentation Dependent on Initial pH by cPCR (cPCR 기법을 이용한 초기배양 pH에 의한 반추위 섬유소 분해 박테리아의 부착 및 발효에 관한 연구)

  • Kim, M.S.;Sung, H.G.;Kim, H.J.;Lee, Sang-S.;Chang, J.S.;Ha, J.K.
    • Journal of Animal Science and Technology
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    • v.47 no.4
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    • pp.615-624
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    • 2005
  • The cPCR technique was used to monitor rumen fermentation and attachment of Fibrobacter succinogenes to cellulose at different pH in the in vitro culture medium. The target fragments of 16S rDNA(445 bp) were amplified from genomic DNA of F. succinogenes with specific primers and internal controls(205 bp) were constructed. Cell counts were estimated from the amounts of genomic DNA, which was calculated from cPCR results. F. succinogenes in pH 6.8 and 6.2 showed apparently higher attachment than in pH 5.8 during all incubation time. There were some difference between pH 6.8 and 6.2 in the degree of attachment, but the different was not significant (P>0.05). Cellulose degradation increased in process of incubation time and the increasing rate was higher when initial pH was higher. The pH in culture medium decreased regardless of initial pH in course of incubation time. After 24 h of incubation, medium pH was dropped by 0.24, 0.58 and 0.16 units from original medium pH 6.8, 6.2 and 5.8, respectively. More gas was produced at higher initial pH in the same manner as in cellulose degradation. In summery, Initial pH of rumen culture in vitro significantly influenced cellulose digestion, gas production, pH change and bacterial attachment. Especially, low pH(5.8) resulted in much lower bacterial attachment and fiber digestion compared to higher medium pH.

Archaeal Diversity in Tidal Flat Sediment as Revealed by 16S rDNA Analysis

  • Kim Bong Soo;Oh Huyn Myung;Kan Ho Jeong;Chun Jong Sik
    • Journal of Microbiology
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    • v.43 no.2
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    • pp.144-151
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    • 2005
  • During the past ten years, Archaea have been recognized as a widespread and significant component of marine picoplankton assemblages. More recently, the presence of novel archaeal phylogenetic lineages has been discovered in coastal marine environments, freshwater lakes, polar seas, and deep-sea hydrothermal vents. Therefore, we conducted an investigation into the archaeal community existing in tidal flat sediment collected from Ganghwa Island, Korea. Phylogenetic analysis of archaeal 16S rDNA amplified directly from tidal flat sediment DNA revealed the presence of two major lineages, belonging to the Crenarchaeota ($53.9\%$) and Euryarchaeota ($46.1\%$) phyla. A total of 102 clones were then sequenced and analyzed by comprehensive phylogenetic analysis. The sequences determined in our samples were found to be closely related to the sequences of clones which had been previously obtained from a variety of marine environments. Archaeal clones exhibited higher similarities ($83.25 - 100\%$) to sequences..from other environments in the public database than did those ($75.22 - 98.46\%$) of previously reported bacterial clones obtained from tidal flat sediment. The results of our study suggest that the archaeal community in tidal flat sediment is remarkably diverse.

Relative Effect of Glyphosate on Glyphosate-Tolerant Maize Rhizobacterial Communities is Not Altered by Soil Properties

  • Barriuso, Jorge;Mellado, Rafael P.
    • Journal of Microbiology and Biotechnology
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    • v.22 no.2
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    • pp.159-165
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    • 2012
  • The rhizobacterial composition varies according to the soil properties. To test if the effect of herbicides on the rhizobacterial communities of genetically modified NK603 glyphosate-tolerant maize varies according to different soil locations, a comparison was made between the effects of glyphosate (Roundup Plus), a post-emergence applied herbicide, and a pre-emergence applied herbicide (GTZ) versus untreated soil. The potential effect was monitored by direct amplification, cloning, and sequencing of the soil DNA encoding 16S rRNA, and high-throughput DNA pyrosequencing of the bacterial DNA coding for the 16S rRNA hypervariable V6 region. The results obtained using three different methods to analyze the herbicide effect on the rhizobacterial communities of genetically modified NK603 maize were comparable to those previously obtained when glyphosate-tolerant maize was grown in soil with different characteristics. Both herbicides decreased the bacterial diversity in the rhizosphere, with Actinobacteria being the taxonomic group most affected. The results suggest that both herbicides affected the structure of the maize rhizobacterial community, but glyphosate was environmentally less aggressive.

Comparative Study of Soil Bacterial Populations in Human Remains and Soil from Keundokgol Site at Buyeo (부여 큰독골 유적 출토 인골 조직 및 외부 토양의 세균 군집의 비교연구)

  • Kim, Yun-ji;Kim, Sue-hoon;Kwon, Eun-sil;Cho, Eun-min;Kang, So-yeong
    • Korean Journal of Heritage: History & Science
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    • v.47 no.4
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    • pp.92-105
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    • 2014
  • Microbial characteristics of bacterial population were investigated in human remains and soil inside the bones in excavated grave no.4 and no.5 at Keundokgol site, Osu-ri, Buyeo. Phylogenetic characteristics of bacterial populations were analyzed by direct extracting of ancient DNA. In this study, based on the 16S rDNA sequences, in case of grave no.4, 319s from human remain were classified into 11 phyla, and 462s from soil were classified into 16 phyla. In case of grave no.5, 271s from human remain were classified into 10 phyla, and 497s from soil were classified into 11 phyla. Especially, Actinobacteria phylogenetic group are dominant group of bacterial populations in grave no.4 and no.5. Also, most of these were analyzed uncultured group. Thus, the discovery of a diversely microbial community and uncultured group was thought to be due to the specificity of the sample. Conclusively the general excavated human bones were contaminated with soil bacteria species their near around. This results contribute to preservation and management of ancient human bone from archaeological sites.

Identification of bacteria from the peri-implant sulcus of orthodontic mini-implants using 16S rDNA clone library (16S rDNA 클론 library 제작 및 핵산염기서열 결정을 통한 교정용 미니임플랜트 주위 열구의 세균 동정)

  • Lim, Sung-Hoon;Kim, Kwang-Won;Yoo, So-Young;Kook, Joong-Ki;Chang, Young-Il
    • The korean journal of orthodontics
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    • v.36 no.4
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    • pp.251-262
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    • 2006
  • Objective: The purpose of this study was to compare the bacterial flora at the peri-implant sulcus of the orthodontic mini-implant placed in the alveolar mucosa with the bacterial flora at the adjacent healthy gingival sulcus. Methods: Two plaque samples from 7 patients were collected by inserting paper points into the sulcus between the mini-implant and ligature wire connected to the mini-implant head and inflamed alveolar mucosa, and from the gingival sulcus of a healthy tooth adjacent to the mini-implant. Results: Using 16S rDNA clone library, the 24 kinds of bacteria including Haemophilus aphrophilus, Sphingomonas species, Capnocytophaga species, Prevotella melaninogenica, Lachnospiraceae species, Porphyromonas species, Neisseria flava were identified only from the sulcus around the mini-implant. These bacteria constituted only 9.2% of total clones, and the bacteria identified from both the sulcus around mini-implants and the gingival sulcus constituted 80.4% of total clones. Of these bacteria, clones of Prevotella species, Atopobium rimae, Veillonella species, Streptococcus intermedius/constellatus, Streptococcus salivarius were more frequently isolated from the peri-implant sulcus. Conclusion: This study suggests that a broad epidemiological study is needed to find causative bacteria which induce inflammation from the peri-implant sulcus.