• Title/Summary/Keyword: 16S rDNA analysis

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Identification of putative periodontal pathogens in Korean chronic periodontitis patients (한국인 만성 치주염 환자에서 치주질환 원인균의 동정)

  • Yun, Jeong-Ho;Park, Jung-Eun;Kim, Doo-Il;Lee, Syng-Ill;Choi, Seong-Ho;Cho, Kyoo-Sung;Lee, Dae-Sil
    • Journal of Periodontal and Implant Science
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    • v.38 no.2
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    • pp.143-152
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    • 2008
  • Purpose: Specific bacteria are believed to play an important role in chronic periodontitis. Although extensive microbial analyses have been performed from subgingival plaque samples of periodontitis patients, systemic analysis of subingival microbiota has not been carried out in a Korean population so far. The purpose of this study was to investigate the prevalence of 29 putative periodontal pathogens in Korean chronic periodontitis patients and evaluate which pathogens are more associated with Korean chronic periodontitis. Material and Methods: A total of 86 subgingival plaque samples were taken from 15 chronic periodontits(CP) patients and 13 periodontally healthy subjects in Korea. CP samples were obtained from the deepest periodontal pocket (>3 mm probing depth[PD]) and the most shallow periodontal probing site ($\leq$3 mm PD) in anterior tooth and posterior tooth, respectively, of each patient. Samples in healthy subjects were obtained from 1 anterior tooth and 1 posterior tooth. Polymerase chain reaction (PCR) of 16S ribosomal DNA (rDNA) of subgingival plaque bacteria was performed. Detection frequencies(% prevalence) of 29 putative periodontal pathogens were investigated as bacterium-positive sites/total sites. Results: With the exception of Olsenella profuse and Prevotella nigrescens, the sites of diseased patients generally showed higher prevalence than the healthy sites of healthy subjects for all bacteria analyzed. Tanerella forsythensis (B.forsythus), Campylobacter rectus, Filifactor alocis, Fusobacterium nucleatum, Porphyromonas endodontalis and Porphyromonas gingivalis were detected in more than 80% of sites with deep probing depths in CP patients. In comparison between the sites (deep or shallow PD) of CP patients and the healthy sites of healthy subjects, there was statistically significant difference(P<0.05) of prevalence in T.forsythensis (B.forsythus), C.rectus, Dialister invisus, F.alocis, P.gingivalis and Treponema denticola. Conclusion: Our results demonstrate that the four putative periodontal pathogens, T.forsythensis (B.forsythus), C.rectus, P.gingivalis and F.alocis are closely related with CP patients in the Korean population.

Effects of Pb and CO2 on Soil Microbial Community Associated with Pinus densiflora-Lab (소나무(Pinus densiflora) 생육토양의 미생물 군집에 미치는 납과 CO2의 영향)

  • Hong, Sun-Hwa;Kim, Sung-Hyun;Kang, Ho-Jeong;Ryu, Hee-Wook;Lee, Sang-Don;Lee, In-Sook;Cho, Kyung-Suk
    • Journal of Ecology and Environment
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    • v.29 no.6
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    • pp.551-558
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    • 2006
  • Effects of Pb and $CO_2$ on soil microbial community associated with Pinus densiflora were investigated using community level physiological profiles (CLPP) and 16S rDNA PCR-denaturing gradient gel electrophoresis (DGGE) methods. Two-years pine trees were planted in Pb-contaminated soils and uncontaminated soils, and cultivated for 3 months in the growth chamber where $CO_2$ concentration was controlled at 380 or 760 ppmv. The structure of microbial community was analyzed in 6 kinds of soil samples (CA-0M : $CO_2$ 380 ppmv + Pb 0 mg/kg + initial, CB-0M : $CO_2$ 380 ppmv + Pb 500 mg/kg + initial, CA-3M : $CO_2$ 380 ppmv + Pb 0 mg/kg + after 3 months, CB-3M : $CO_2$ 380 ppmv + Pb 500 mglkg + after 3 months, EA-3M : $CO_2$ 760 ppmv + Pb 0 mg/kg + after 3 months, EB-3M : $CO_2$ 760 ppmv + Pb 500 mg/kg + after 3 months). After 3 months, the substrate utilization in the uncontaminated soil samples (CA-3M vs EA-3M) was not significantly influenced by $CO_2$ concentrations. However, the substrate utilization in the Pb-contaminated soil samples (CB-3M vs EB-3M) was enhanced by the elevated $CO_2$ concentrations. The results of principal component analysis based on substrate utilization activities showed that the structure of microbial community structure in each soil sample was grouped by Pb-contamination. The similarities of DGGE fingerprints were 56.3 % between the uncontaminated soil samples (CA-3M vs EA-3M), and 71.4% between the Pb-contaminated soil samples (CB-3M vs. EB-3M). The similarities between the soil samples under $CO_2$ 380 ppmv (CA-3M vs CB-3M) and $CO_2$, 760 ppmv (EA-3M vs EB-3M) were 53.3% and 35.8%, respectively. These results suggested that the structure of microbial community associated with Pinus densiflora were sensitively specialized by Pb-contamination rather than $CO_2$ concentration.

Potential Probiotic Properties of Lactobacillus johnsonii IDCC 9203 Isolated from Infant Feces (유아 분변에서 분리한 Lactobacillus johnsonii IDCC 9203의 잠재적 프로바이오틱 특성)

  • Lee, Seung-Hun;Yang, Eun-Hee;Kwon, Hyuk-Sang;Kang, Jae-Hoon;Kang, Byung-Hwa
    • Microbiology and Biotechnology Letters
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    • v.36 no.2
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    • pp.121-127
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    • 2008
  • A strain IDCC 9203 isolated from infant feces was identified as Lactobacillus johnsonii on the basis of 16S rDNA sequence analysis. L. johnsonii IDCC 9203 was highly resistant to acid (MRS broth at pH 2.3) and bile (MRS broth with 0.3% oxgall). The antibacterial activities of L. johnsonii IDCC 9203 was examined against Salmonella typhimurium KCTC 2054. The growth of S. typhimurium KCTC 2054 was inhibited by the cell-free culture supernatant (at pH 4.0) of L. johnsonii IDCC 9203 as well as by the respective control (MRS broth at pH 4.0). Antimicrobial effect against S. typhimurium KCTC 2054 of L. johnsonii IDCC 9203 was probably due to the lactic acid. By an in vitro cell adhesion model, L. johnsonii IDCC 9203 preincubated or coincubated with Caco-2 cells reduced the adhesion of S. typhimurium KCTC 2054 to Caco-2 cells by 74% or 47.1%, respectively. Also in an in vivo model, L. johnsonii IDCC 9203 was colonized in mice intestines which were disrupted by ampicillin treatment. Its proliferation in the mice intestines reduced abnormal salmonella growth from $10^9CFU/g$ feces to $10^5CFU/g$ feces as an indigenous level. The results obtained in this study suggest that L. johnsonii IDCC 9203 may be a potential probiotic strain.

Characteristics of Ju-Back and Effect of Ju-Back Fertilizer on Growth of Crop Plants (주류생산 부산물인 주박의 특성 규명 및 주박이 작물생육에 미치는 영향)

  • Lee, Jung-Hoon;Park, Sung-Min;Park, Chi-Duck;Jung, Hyuck-Jun;Kim, Hyun-Soo;Yu, Tae-Shick
    • Journal of Life Science
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    • v.17 no.11
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    • pp.1562-1570
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    • 2007
  • This experiment was conducted to develop fertilizer which promotes plant growth as well as suppressing pathogenic fungi. The fertilizer was made from the mixture of Ju-Back (Korean rice wine cake) and indigenous rhizosphere-bacterium. The main ingredients of Ju-Back were investigated as 6.04% total nitrogen, 42.59% total carbohydrate, 1.01% available phosphate, 73.42% organic matter, 7.72% potassium oxide, 1.35% calcium oxide, 0.53% magnesium oxide. The enzyme activities of Ju-Back were estimated to be 980 units/g for ${\alpha}-amylase$, 300 units/g for glucoamylase, and 1800 units/g for acid pretense. Indigenous rhizosphere bacteria which produced antifungal agent were isolated from soil, and was selected KMU-13 strain which can antagonize against various plant pathogenic fungi (Botrytis cinerea KACC 40573, Sclerotinia sclerotiorum KACC 41065, Fusairum oxysporum KACC 40052, Pythium aphanidermatum KACC 40156, Phytophthora capsici KACC 40476 and Glomerella cingulata KACC 40299). KMU-13 strain was identified as Bacillus subtilis KMU-13 by biochemical and 16s rDNA analysis. The organic fertilizer was made as prototype which was composed 20% Ju-Back, 70% carrier, 9.7% microorganism cultivated solution, 0.3% trace-element. We also investigated an application of fertilizer using Ju-Back for cultivating lettuce (Lactuca sativar) which were grown in three soil conditions that had chemical fertilizer, barnyard manure, lime power, urea, potassium chloride and superphosphate as a control, the whole quantity (80 kg/10a) of posted fertilizer with the control and the half quantity (40 kg/10a) with the control. The growth characteristics were examined and analysed with several weeks interval from 3 weeks to 8 weeks on head length (cm), head width (cm/head), number of leaf and fresh weight (g/plant). The results are summarized as follows. The head width and fresh weight of lettuce were the highest at posted fertilizer 1 (whole quantity) was applied chemical, organic matter (Ju-Back) and carrier. The head length was the highest at posted fertilizer 2 (whole quantity) was applied Ju-Back only.

Characterization of typical Aeromonas salmonicida isolated from Sea-Chum Salmon (Oncorhynchus keta) (해수에 순치된 첨연어(Oncorhynchus keta)에서 분리된 정형 에로모나스 살모니시다(Aeromonas salmonicida)에 대한 특성 분석)

  • Jongwon Lim;Sungjae Ko;Youngjun Park;Do-il Ahn;Suhee Hong
    • Journal of fish pathology
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    • v.36 no.2
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    • pp.263-275
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    • 2023
  • Chum salmon (Oncorhynchus keta) is a species which returns to Korea for spawning and was produced as seed production at the Fisheries Resources Agency located in Uljin-gun, Gyeongsangbuk-do to preserve the species. However, farmed chum salmon showed symptoms of bacterial infection. Therefore, in this study, bacteria were isolated to identify the causative agent from chum salmon in October 2021. The isolated bacteria were identified based on the sequences of 16S rDNA, rpoD (RNA polymerase sigma factor σ70), and vapA (A-layer) genes. Also, salinity-growth curve, biochemical characterization, antibiotic susceptibility test, and pathogenicity analysis were performed in four strains. As a result, four isolated strains were identified as Aeromonas salmonicida subsp. salmonicida. Additionally, the bacterial strains showed a decrease in growth as the salt concentration increased in the medium. All of the isolated strains exhibited γ-hemolysis, and the same biochemical properties. In the antimicrobial susceptibility test, all strains showed an inhibition zone of 40 to 44 mm for oxolinic acid, flumequine, and florfenicol. Pathogenic factors were assessed by RT-PCR at the mRNA level, and found that the four strains expresses the outer membrane ring of T3SS (ascV), inner membrane ring of T3SS (ascC), vapA, enterotoxin (act), and lipase (lip) genes which are well known to significantly contribute to the pathogenicity of A. salmonicida. The results of this study can be used as basic data to prevent A. salmonicida subsp. salmonicida occurring in sea-chum salmon in the future.

Identification of electron beam-resistant bacteria in the microbial reduction of dried laver (Porphyra tenera) subjected to electron beam treatment (전자선 처리에 따른 마른 김(Porphyra tenera)의 미생물 저감화 효과와 저항성 세균의 동정)

  • Kim, You Jin;Oh, Hui Su;Kim, Min Ji;Kim, Jeong Hoon;Goh, Jae Baek;Choi, In Young;Park, Mi-Kyung
    • Food Science and Preservation
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    • v.23 no.1
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    • pp.139-143
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    • 2016
  • This study investigated the effect of electron beam (EB) treatment on the microbial reduction of dried laver (Porphyra tenera) and identified EB-resistant bacteria from the treated dried laver. After EB treatments of 4 kGy and 7 kGy, the numbers of total bacteria and EB-resistant bacteria were measured using tryptic soy agar and mannitol salt agar, respectively. The morphological and biochemical characteristics of each isolated EB-resistant bacteria were investigated and these bacteria were identified. Compared to the control ($1.5{\pm}0.2){\times}10^6CFU/g$, the total bacterial number was significantly decreased to ($5.4{\pm}0.5){\times}10^4CFU/g$ and ($1.1{\pm}0.6){\times}10^4CFU/g$ after EB treatments of 4 kGy and 7 kGy, respectively. With a higher EB dosage, the number of red colonies was almost same, whereas the number of yellow colonies was significantly decreased to ($3.3{\pm}1.2){\times}10^3CFU/g$ and 0 CFU/g for 4 kGy and 7 kGy, respectively. All red and yellow colonies were gram-positive cocci, catalase-positive, and resistant to 3% and 5% NaCl media. From the 16S rDNA sequence analysis, yellow and red colonies were identified as either Micrococcus flavus or M. luteus, with 99% similarity for the yellow colonies, and Deinococcus proteolyticus and D. piscis, with 99% and 97% similarity for the red colonies, respectively.

Expressed sequence tag analysis of Meretrix lusoria (Veneridae) in Korea (한국산 백합 (Meretrix lusoria) 의 전사체 분석)

  • Kang, Jung-Ha;Jeong, Ji Eun;Kim, Bong Seok;An, Chel-Min;Kang, Hyun-Sook;Kang, Se-Won;Hwang, Hee Ju;Han, Yeon Soo;Chae, Sung-Hwa;Ko, Hyun-Sook;Lee, Jun-Sang;Lee, Yong Seok
    • The Korean Journal of Malacology
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    • v.28 no.4
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    • pp.377-384
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    • 2012
  • The importance of biological resources has been gradually increasing, and mollusks have been utilized as main fishery resources in terrestrial ecosystems. But little is known about genomic and transcriptional analysis in mollusks. This is the first report on the transcriptomic profile of Meretrix lusoria. In this study, we constructed cDNA library and determined 542 of distinct EST sequences composed of 284 singletons and 95 contigs. At first, we identified 180 of EST sequences that have significant hits on protein sequences of the exclusive Mollusks database through BLASTX program and 343 of EST sequences that have significant hits on NCBI NR database. We also found that 211 of putative sequences through local BLAST (blastx, E < e-10) search against KOG database were classified into 16 functional categories. Some kinds of immune response related genes encoding allograft inflammatory factor 1 (AIF-1), B-cell translocation gene 1 (BTG1), C-type lectin A, thioester-containing protein and 26S proteasome regulatory complex were identified. To determine phylogenetic relationship, we identified partial sequences of four genes (COX1, COX2, 12S rRNA and NADH dehydrogenase) that significantly matched with the mitochondrial genomes of 3 species-Ml (Meretrix lusoria), Mp (Meretrix petechialis) and Mm (Meretrix meretrix). As a result, we found that there was a little bit of a difference between sequences of Korean isolates and other known isolates. This study will be useful to develop breeding technology and might also be helpful to establish a classification system.

Purification and Characterization of Bacillus subtilis JS-17 Collagenase. (Bacillus subtilis JS-17이 생산하는 Collagenase의 정제 및 특성)

  • Lim Kyoung-Suk;Son Shung-Hui;Kang Ho Young;Jun Hong-Ki
    • Journal of Life Science
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    • v.15 no.4 s.71
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    • pp.657-663
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    • 2005
  • Collagenases are generally defined as enzymes that are capable of degrading the polypeptide backbone of native collagen under conditions that do not denature the protein. An extracellular collagenase-producing bacterial strain was isolated from kimchi and identified to be Bacillus subtilis JS-17 through morphological, cultural, biochemical characteristics and 16S rDNA sequence analysis. Optimum culture condition of Bacillus subtilis JS-17 for the production of collagenase was $1.5\%$ fructose, $1\%$ yeast extract, $0.5\%\;K_2HPO_4,\;0.4\%\;KH_2PO_4,\;0.01\%\;MgSO_4\cdot7H_2O,\;0.01\%\; MnSO_4\cdot4H_2O,\;,0.1\%$ citrate and $0.1\%\;CaCl_2$. The production of collagenase was optimal at $30^{\circ}C$ for 72 hr. A collagenase was isolated from the culture filtrate of Bacillus subtilis JS-17. The enzyme was purified using Amberlite IRA-900 column chromatography, Sephacryl S-300 HR column chromatography and DEAE-Sephadex A-50 column chromatography The purified collagenase has an specific activity 192.1 units/mg. The molecular weight of the purified enzyme was estimated to be 28 kDa by SDS-PACE. The purified collagenase has $100\%$ activity up to $55^{\circ}C$.

A Major Locus for Quantitatively Measured Shank Skin Color Traits in Korean Native Chicken

  • Jin, S.;Lee, J.H.;Seo, D.W.;Cahyadi, M.;Choi, N.R.;Heo, K.N.;Jo, C.;Park, H.B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.11
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    • pp.1555-1561
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    • 2016
  • Shank skin color of Korean native chicken (KNC) shows large color variations. It varies from white, yellow, green, bluish or grey to black, whilst in the majority of European breeds the shanks are typically yellow-colored. Three shank skin color-related traits (i.e., lightness [$L^*$], redness [$a^*$], and yellowness [$b^*$]) were measured by a spectrophotometer in 585 progeny from 68 nuclear families in the KNC resource population. We performed genome scan linkage analysis to identify loci that affect quantitatively measured shank skin color traits in KNC. All these birds were genotyped with 167 DNA markers located throughout the 26 autosomes. The SOLAR program was used to conduct multipoint variance-component quantitative trait locus (QTL) analyses. We detected a major QTL that affects $b^*$ value (logarithm of odds [LOD] = 47.5, $p=1.60{\times}10^{-49}$) on GGA24 (GGA for Gallus gallus). At the same location, we also detected a QTL that influences $a^*$ value (LOD = 14.2, $p=6.14{\times}10^{-16}$). Additionally, beta-carotene dioxygenase 2 (BCDO2), the obvious positional candidate gene under the linkage peaks on GGA24, was investigated by the two association tests: i.e., measured genotype association (MGA) and quantitative transmission disequilibrium test (QTDT). Significant associations were detected between BCDO2 g.9367 A>C and $a^*$ ($P_{MGA}=1.69{\times}10^{-28}$; $P_{QTDT}=2.40{\times}10^{-25}$). The strongest associations were between BCDO2 g.9367 A>C and $b^*$ ($P_{MGA}=3.56{\times}10^{-66}$; $P_{QTDT}=1.68{\times}10^{-65}$). However, linkage analyses conditional on the single nucleotide polymorphism indicated that other functional variants should exist. Taken together, we demonstrate for the first time the linkage and association between the BCDO2 locus on GGA24 and quantitatively measured shank skin color traits in KNC.

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.