• Title/Summary/Keyword: mycobacteria

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Isolation of Environmental Mycobacteria from Diverse Water Samples Using Cetylpyridinium Chloride

  • Choi, Yeon-Im;Jin, Hyun-Woo;Lee, Gyu-Sang;Kim, Jong-Bae;Song, In-Kenn;Kim, Young-Joon;Lee, Hye-Young
    • Biomedical Science Letters
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    • v.16 no.3
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    • pp.133-138
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    • 2010
  • Despite of the increasing importance of environmental mycobacteria, detection and identification of mycobacteria from environmental sources including water have been fraught with technical difficulties. Although, several protocols to optiruize isolation of mycobacteria from water sources have been reported, standard method has not yet been established. In this study, usefulness of cetylpyridinium chloride (CPC), a cationic quaternary ammonium compound, for the isolation of environmental mycobacteria from diverse water samples was evaluated. For this, water samples from diverse water sources such as effluent water, lake water, and underground water were collected, treated with diverse concentrations of CPC, and plated on the solid agar plates. Subsequently individual colonies grown on the plates were sequence analyzed for identification of each colony. In brief, the results from this study showed that the growth of mycobacteria was enhanced by use of CPC as a pre-treatment reagent to water samples by inhibiting growth of other non-mycobacteria in water. In fact, the effect of CPC to decontaminate non-mycobacteria for isolation of mycobacteria was better than 1~4% of NaOH, which is a routinely used decontaminating reagent widely employed for culturing mycobactera from sputum specimens. Therefore, the results from this study seems to support that the CPC pre-treatment may be useful for isolation of mycobacteria from diverse sources including clinical specimens which are often contaminated with other bacteria.

Synthesis of ADD and AD by Mycobacteria sp. in Microemulsions (Microemulsion에서 Mycobacteria를 이용한 ADD와 AD의 합성)

  • 이강민;김영득김희정박충웅
    • KSBB Journal
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    • v.7 no.3
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    • pp.161-165
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    • 1992
  • Water-insoluble sitosterol was biotransformed to AD(4-androstene-3, 17-dione) and ADD(1, 4-androstadiene-3, 17-dione) with crosslinked Mycobacteria sp. NRRL B-3683 in microemulsions. The Mycobacteria activity depended on the kinds of surfactants and water content. The activity was very high in cationic microemulsion (CTAB-buthanol-cyclohexane-water) and five times higher than that of buffer, but it showed a very low activity in anionic microemulsion. The Mycobacteria activity was increased as increasing as increasing water contents and in the microemulsion containing 20% water was doubled in the same microumulsion containing 5% water. We suggest that microemulsion is effective reaction medium for Mycobacteria bioconversion.

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Occurrence and Molecular Differentiation of Environmental Mycobacteria in Surface Waters

  • Lee, Eun-Sook;Lee, Mok-Young;Han, Sun-Hee;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • v.18 no.7
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    • pp.1207-1215
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    • 2008
  • To investigate the occurrence and species diversity of mycobacteria in waters, surface water samples were collected monthly from the Han River and tap water samples at the terminal sites of the distribution system. Mycobacteria in each water sample were isolated by decontamination using cetylpyridinium chloride (CPC) and cultivation on Middlebrook 7H10 agar, and then identified by polymerase chain reaction-restriction fragment length polymorphism analysis (PRA) and sequencing of the 65-kDa heat-shock protein gene (hsp65 gene). Mycobacteria were detected in 59% of the surface water samples and 26% of the tap water samples. Over half of the 158 isolates could not be identified by hsp65 PRA and gene sequencing, and several identification discrepancies were observed between the two methods. The most frequently isolated species was Mycobacterium gordonae in surface water and M. lentiflavum in tap water. M. avium complex (MAC), the most important pathogen among environmental mycobacteria, was detected in the surface water samples but not found in the tap water samples. The result demonstrated that water is an important environmental source of mycobacteria and the combined application of hsp65 PRA and sequencing was more reliable than hsp65 PRA alone to accurately identify mycobacteria present in water.

Identification of Mycobacteria Using Polymerase Chain Reaction and Sputum Sample (객담을 이용한 Mycobacteria의 검출과 중합효소 연쇄반응의 민감성 비교)

  • Jang, Hyung Seok
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.2
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    • pp.83-89
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    • 2015
  • Although Mycobacterium tuberculosis complex strains remain responsible for the majority of diseases caused by mycobacterial infections worldwide, the increase in HIV (human immuno deficiency virus) infections has allowed for the emergence of other non-tuberculous mycobacteria as clinically significant pathogens. M. tuberculosis was detected by two-tube nested polymerase chain reaction (PCR) and non-tuberculous mycobacteria was detected by PCR-restriction fragment length polymorphism (RFLP) with Msp I. Result of niacin test is equal to result of two-tube nested PCR after culture for M. tuberculosis. In this study, acid fast bacilli stain (AFB. stain) >2+ case, Detection of Mycobacteria is similar to result before culture and after culture. AFB. stain <1+ case, result of mycobacteria is distinguished. Conclusionly, these results suggest that identification of mycobacteria must go side by side both culture and PCR for more fast and accuracy.

Antimicrobial Peptides in Innate Immunity against Mycobacteria

  • Shin, Dong-Min;Jo, Eun-Kyeong
    • IMMUNE NETWORK
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    • v.11 no.5
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    • pp.245-252
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    • 2011
  • Antimicrobial peptides/proteins are ancient and naturally-occurring antibiotics in innate immune responses in a variety of organisms. Additionally, these peptides have been recognized as important signaling molecules in regulation of both innate and adaptive immunity. During mycobacterial infection, antimicrobial peptides including cathelicidin, defensin, and hepcidin have antimicrobial activities against mycobacteria, making them promising candidates for future drug development. Additionally, antimicrobial peptides act as immunomodulators in infectious and inflammatory conditions. Multiple crucial functions of cathelicidins in antimycobacterial immune defense have been characterized not only in terms of direct killing of mycobacteria but also as innate immune regulators, i.e., in secretion of cytokines and chemokines, and mediating autophagy activation. Defensin families are also important during mycobacterial infection and contribute to antimycobacterial defense and inhibition of mycobacterial growth both in vitro and in vivo. Hepcidin, although its role in mycobacterial infection has not yet been characterized, exerts antimycobacterial effects in activated macrophages. The present review focuses on recent efforts to elucidate the roles of host defense peptides in innate immunity to mycobacteria.

Thin-layer chromatography of mycobacterial lipids as an aid to classification - I. Reference strains (박층(薄層) 크로마토그라피(Thin-layer chromatography)에 의(依)한 항산균(抗酸菌)의 지방질(脂肪質)을 이용(利用)한 분류(分類) - I. 표준균주(標準菌株)에 대(對)하여)

  • Koh, Choon-Myung;Kim, Sung-Kwang;Choi, Tae-Kyung;Pyun, Woo-Sup
    • The Journal of the Korean Society for Microbiology
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    • v.8 no.1
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    • pp.33-36
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    • 1973
  • This study was carried out for the classification of mycobacteria using the thin-layer chromatography of mycobacterial lipids. Results as follows: Of the 12 strains of mycobacteria, the two spots on chromatogarphy were three strains of mycobacteria(BCG, M. tuberculosis($H_{37}J$) and M. ulcerans) and three spots on chromatography were two strains of mycobacteria(M. kansasii and M. balnei). The method of thin-layer chromatography of mycobacterial lipids was considered to capable for the use of classification of mycobacteria.

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Genetic Mutations of rpoB of Mycobacteria Resistance to Rifampin (Rifampin 내성 마이코박테리아의 rpoB 유전자 변이)

  • Kweon, Tae-Dong;Sa, Young-Hee;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2012.05a
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    • pp.913-915
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    • 2012
  • RNA polymerase beta subunit gene (rpoB) mutation of rifampin-resistant Mycobacteria was analyzed using nucleotide sequence of rpoB DNA (351 bp) containing rifampin resistant region, $rif^r$. For this purpose, we collected rifampin-resistant Mycobacteria that were identified by conventional culture method from Masan National Hospital and The Korean Institute of Tuberculosis and performed analysis of nucleotide sequence of rpoB of them. We found various mutations of rpoB linked rifampin resistant gene from rifampin-resistant Mycobacteria. From this study, we identified mutations of different codons from codons that have been reported recently.

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Subdivision of Opportunist Mycobacteria by the Difference of Pigment Production on Lowenstein-Jensen Medium Containing Crocin (Crocin첨가 결핵균 배지상의 색소형성에 의한 비정형 Mycobacteria의 분류)

  • Choi, Chul-Soon;Kim, Jae-Hak;Yoon, Yong-Dhuk;Lee, Hyun-Soo;Lee, Taek-Chu
    • The Journal of the Korean Society for Microbiology
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    • v.9 no.1
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    • pp.33-40
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    • 1974
  • On the basis of pigment production and growth rate on L-J medium containing crocin, differentiation of opportunist mycobacteria belonging to photochromogens, scotochromogens, nonchromogens and rapid grower has been investigated. Among photochromgens, positive pigmentation of M. kansasii was differentiated from negative strain of M. marinum, Scotochromogen M. aquae was positive whereas M. scrofulaceum was negative. Rapid grower M. fortuitum was positive at 3 days test whereas M. smegmatis was negative. Subdivision of opportunist mycobacteria into four groups on the basis of growth rate and pigment production on L J medium containing gardenia extraction appeared to be a valuable adjunct to the Runyon's classification for the rapid presumptive identification of opportunist mycobacteria of different clinical significance.

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Diverse Mutations of rpoB in Rifampin-Resistant Mycobacteria (Rifampin에 대한 내성 마이코박테리아에서 rpoB의 다양한 변이)

  • Kweon, Tae-Dong;Sa, Young-Hee;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2012.10a
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    • pp.991-993
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    • 2012
  • We analyzed RNA polymerase beta subunit gene (rpoB) mutation of rifampin-resistant Mycobacteria through analysis of nucleotide sequence of rpoB DNA (351 bp) containing rifampin resistant region, $rif^r$. For this study, we collected rifampin-resistant Mycobacteria that were identified by conventional culture methods from Masan National Hospital and The Korean Institute of Tuberculosis. We performed sequencing of DNA nucleotides and analyzed rpoB gene of those rifampin-resistant Mycobacteria. From this analysis, we invcestigated diverse mutations of rpoB gene included rifampin-resistant gene, which were not reported, from those rifampin-resistant Mycobacteria.

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Isolation and Identification of Opportunist Mycobacteria and Nocardia from Soil Specimens of School Ground in Seoul City (서울시내(布內) 초중고등학교(初中高等學校) 토양(土壤)으로부터 비정형(非定型) Mycobacteria와 Nocardia의 분리(分離) 및 생화학적(生化學的) 동정(同定))

  • Choi, Chul-Soon;Yang, Yong-Tae
    • The Journal of the Korean Society for Microbiology
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    • v.11 no.1
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    • pp.69-78
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    • 1976
  • There have been increasing reports of mycobacterioses in man and animals caused by "atypical" or "opportunist" mycobacteria. At the presnt, "opportunist mycobacterioses" are not generally responsive to antituberculosis drugs, and therefore, create considerable problems with regard to chemotherapy and control measures. In recent years studies have been made to isolate opportunist mycobacteria from soil, house-dusts and tap-water. It seemed quite interesting to define the extent of circumstantial presence of "opportunist" mycobacteria and nocardia in the soils of school-ground of primary schools and middle-high schools. This communication is the results of pilot study to isolate and identify "opportunist" mycobacteria and nocardia from 504 soil specimens of 72 schools in Seoul City. 1. Of a total of 59 isolates from 504 soil specimens tested, 32 strains were identified as opportunist mycobacteria and 27 strains as nocardia. 2. Isolation rates of opportunist mycobacteria by the areas(of specimen collection) were as follows: 36.4% in the southern area of Han-River, 33.3% in the central area, 22.7% in the outskirt area and 16.6% in the intermediate area. There observed no apparent difference in the isolation rates both-between the areas and between primary schools and middle-high schools. However, a significant difference was noted in the isolation rates between the places of soil sampling in a given school(P<0.05), i.e., the highest was the soil of refuge heaps(15.2%), and tap-water pole area(11.1%), the school-lavatory entrance(9.7%), the school gate entrance(5.5%), and iron-bar play ground(2.7%). The soil specimens from the center of school ground and from school building entrance yielded none of mycobacterial isolates. 3. Isolation rates of nocardia by the areas were as follows: 33.3% in the central area, 31.8% in the outskirt area, 27.3% in the southern ares of Han-River and 11.1% in the intermediate area. As in the case of mycobacteral isolates, there observed no apparent differences in the isolation rates both between the areas and between primary schools and middle-high schools, but a significant difference was noted between the places of soil sampling(P<0.05), i.e., the highest was the soil of school building entrance(15.2%), and of school gate entrance(6.9%), refuge heaps(5.5,%), iron-bar play ground(4.1%), the school-lavatory entrance(2.7%) and tap-water pole area(2.7%), respectively. The soil specimens from the center of, school ground yielded none of nocardia isolates. 4. Of the 32 strains of isolated mycobacteria. 15 strains were slow-growing mycobacteria and the remaining 17 strains belonged to the rapid growers. Of the 15 slow-growers. 4 strains were M. scrofulaceum-szulgai complex, 3 M. gordonae, 4 M. terrae-triviale complex, 2 M. avium-intracellulare-xenopi complex, and 2 unclassified schotochromogens. Of the 17 strains of rapid growers, 12 were M. diernhoferi, 2 M. fortuitum-peregrinum complex, 2 M. vaccae and one M. flavescens. 5. Of the 27 strains of nocardia isolated, 11 strains were N. transvalensis, 5 N. convoluta, 5 N. erythropolis, one N. vaccinii, one N. polychromogens-paraffinae complex and 4 untypable strains of orange-pigmented nocardia spp.

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