• Animal cells and systems
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• v.4 no.3
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• pp.299-304
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• 2000

In the present paper, the immunostimulatory effects of the extracellular products (ECP) from Mycobacterium spp. and various adjuvants on the non-specific immune responses of Nile tilapia, Oreochromis nilotica, were examined. Nile tilapia were immunized by injecting ECP of Mycobacterium spp. (strain TB40, TB267 or the type strain Mycobacterium marinum) into their swim bladders. A variety of adjuvants like as Freund s complete adjuvant (FCA), Freund's incomplete adjuvant (FIA) and Titremax were similarly injected into additional groups of tilapia. The number of nitroblue tetrazolium (NBT)-positive cells observed in the swim bladder of the immunized fish was signigicantly increased by the fourth day post-immunization. By day 8, the numbers of NBT-positive cells were fewer in fish immunized with ECP from mycobacteria strains TB40 or TB267 than those immunized with ECP from M. marinum or fish injected with FCA or FIA. The level of Iysozyme activity detected in the serum of fish 40 alter immunization with ECP from various Mycobacterium spp. was also significantly higher than that found in the serum of the control fish. Head kidney macrophages showed enhanced reduction of NBT when cultured in vitro with 1 $\mu$ g/ml of ECP. Concentrations greater than this (10 or 100 $\mu$g/ml) were found to suppress the reduction of NBT by the macrophages. ECP from Mycobacterium spp. and the various adjuvants used in the study all appear to be good activators of the non-specific immune responses of Nile tilapia.

• Journal of Life Science
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• v.20 no.4
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• pp.578-583
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• 2010

It has been reported that extracts of globe thistle (Echinops spp.) and thistle (Circium spp., Carduus spp. and Onopordum spp.) have anti-bacterial and anti-fungal activities. Methanol extracts of Echinops setifer and Carduus spp. were used to test and see if the extracts of these plants could suppress growth of Mycobacterium smegmatis and Mycobacterium fortuitum. Although extract of Echinops setifer showed no anti-mycobacterial activities, extract of Carduus spp. showed inhibition zones when tested with filter discs. Genomic DNA was isolated from Carduus spp. and PCR was performed to clone a DNA fragment containing ITS1, 5.8S rRNA gene and ITS2. A 733-bp PCR product was obtained and its DNA sequence was reported to the GenBank (accession number GU188570). BLAST search of the obtained DNA sequence did not show a match with any DNA sequences in the Genbank. Carduus crispus and Carduus defloratus had the closest phylogenetic relationships to this plant.

• Biomedical Science Letters
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• v.16 no.1
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• pp.10-18
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• 2010

The present study was set to develop comprehensive system for assessing the safety of drinking water using PCR-reverse blot hybridization assay (REBA). The REBA developed in this study can detect waterborne pathogens such as Shigella spp., Salmonella spp., Citrobacter spp., Enterobacter spp., Klebsiella spp., Yersinia spp., Mycobacterium spp., Listeria spp. at the genus level, and Escherichia coli, Citrobacter freundii, Klebsiella pneumoniae, Pseudomonas aeruginosa, Yersinia enterocolitica, Y. pseudotuberculosis, Mycobacterium avium complex, M. marinum, Enterococcus faecalis, and Staphylococcus aureus at the species level, and E. coli O157:H7 at the strain level.

• Korean Journal of Veterinary Service
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• v.30 no.2
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• pp.233-241
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• 2007

Tuberculin positive cattle without gross tubercle lesions should be confirmed by the bacteriological examination to determine the state of the infection. To overcome the time-consuming and laborious identification by culture and biochemical tests, polymerase chain reaction (PCR) has been used to identify Mycobacterium bovis. Due to various lipids in the cell wall of Mycobacterium spp, novel methods of DNA extraction from Mycobacterium spp have been developed. In this study, a newly developed guanidium isothiocyanate/silica DNA extraction method was directly applied to specimens from the tuberculin positive cattle. DNAs were directly extracted from the lymph nodes and the major polymorphic tandem repeat (MPTR) and mycobacterial protein of BCG 70 (MPB70) were amplified using PCR. The DNA extraction method using guanidium isothiocyanate/silica was efficient and safe, and the MPTR and MPB70 primers were specific to M bovis. Therefore, MPTR and MPB70 PCRs will be useful for the detection of M bovis in the lymph node from skin-test positive cattle.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.12 no.8
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• pp.3517-3522
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• 2011

Contamination of public bath water by waterborne pathogens can cause disease outbreaks and contribute to background rates of disease. The aim of this study is to determine the prevalence of waterborne pathogens in public baths. A total of 30 water samples were collected from 30 different public baths in seoul, Korea. Pathogens in water samples were concentrated by 0.45 ${\mu}m$ nitrocellulose membrane filter, analyzed by both cultivation and polymerase chain reaction-reverse blot hybridization (PCR-REBA) of partial 16S rRNA gene. Various microorganisms including Escherichia coli and Shigella spp. were identified by microbiological cultivation. E. coli, Shigella spp., Salmonella spp., Pseudomonas spp. and Mycobacterium spp. were identified by PCR-REBA. Our results suggest that appropriate hygiene practice and continuous monitoring is needed for reducing health risk associated with public bath houses.

• Korean Journal of Veterinary Research
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• v.29 no.3
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• pp.283-289
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• 1989

In order to measure in vitro cell mediated immunity in the guinea pigs sensitized with the killed bacilli of Mycobacterium bovis ($AN_5$), M avium (serotype 2), M tuberculosis and M intracellulare (serotype 8), leukocyte adherence inhibition (LAI) test was established using the antigens of purified protein derivatives (PPD) tuberculin. By using LAI test, specificity of cell-mediated immune responses of the guinea pigs inoculated with various Mycobacterium spp was investigated, and comparison between values of LAI and skin test was also made to evaluate the specificity of the newly designed test. The results obtained throughout the experiments were summarized as follows; 1. The optimal concentration of PPD antigens for LAI test was 1 to 2mg per ml of medium. 2. When the leukocytes of guinea pigs sensitized with both M bovis($AN_5$) and M avium (serotype 2) for 2 to 8 weeks were incubated with homologous or heterologous PPD antigens, mean values of LAI test were $61.2{\pm}11.2$ and $65.6{\pm}5.1%$ in homologous PPD antigens respectively, while $30.0{\pm}3.7$ and $32.8{\pm}5.7%$ in heteNlogous PPD antigens, showing the prominently high value of LAI in the homologous syst,em (p<0.01). 3. When the leukocytes of guinea pigs sensitized with both M tuberculosis and M intracellulare (serotype 8) for 2 to 8 weeks were incubated with homologous and heterologous PPD antigens, mean values of LAI test were $67.9{\pm}2.9$ and $66.9{\pm}5.0%$ in homologous PPD antigens, while $27.4{\pm}7.4$ and $24.4{\pm}7.1%$ in heterologous PPD antigens, showing the prominently high value of LAI in the homologous system (p<0.01). 4. Comparing with the specificity of LAI and skin tests on the basis of the value obtained from the homologous system, deviation of reaction was revealed to be 49.5 to 100.2 in LAI test, and -15.9 to 52.0 in skin test.

• Parasites, Hosts and Diseases
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• v.45 no.1 s.141
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• pp.11-18
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• 2007

Recent in vitro studies have revealed that a certain Mycobacterium can survive and multiply within free-living amoebae. It is believed that protozoans function as host cells for the intracellular replication and evasion of Mycobacterium spp. under harmful conditions. In this study, we describe the isolation and characterization of a bacterium naturally observed within an amoeba isolate acquired from a contact lens storage case. The bacterium multi-plied within Acanthamoeba, but exerted no cytopathic effects on the amoeba during a 6-year amoebic culture. Trasnmission electron microscopy showed that the bacteria were randomly distributed within the cytoplasm of trophozoites and cysts of Acanthamoeba. On the basis of the results of 18S rRNA gene analysis, the amoeba was identified as A. lugdunensis. A 16S rRNA gene analysis placed this bacterium within the genus Mycobacterium. The bacterium evidenced positive reactivity for acid-fast and fluorescent acid-fast stains. The bacterium was capable of growth on the Middlebrook 7H11-Mycobacterium-specific agar. The identification and characterization of bacterial endosymbionts of free-living protozoa bears significant implications for our understanding of the ecology and the identification of other atypical mycobacterial pathogens.

• Korean Journal of Clinical Laboratory Science
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• v.51 no.2
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• pp.177-184
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• 2019

Mycobacteria grow slowly. Therefore, a solid medium should be used for eight weeks and a liquid medium for six weeks. The purpose of this study was to find the growth factors that can grow Mycobacterium rapidly and to help develop a solid medium for rapid identification. Three types of Mycobacterium growth factors were evaluated with 10 Mycobacteria by adding activated charcoal, defibrinated sheep blood, and L-ascorbic acid to $Difco^{TM}$ Mycobacteria 7H11 agar (Becton, Dickinson and Company, Sparks, MD, USA). The time to detection and the distinguishability of a colony were compared with that of the current method. In the rapidly growing Mycobacterium, the difference in detection time between the new media and conventional media confirmed that the new media was faster. M. kansasii and M. intracelluare grew faster in 7H11 C than in 7H11 medium. MTB grew faster than the other media in 7H11 C. This study confirmed that the two growth factors affect fast-growing Mycobacteria and slow-growing Mycobacteria. 7H11 C showed better distinguishability than the conventional media in all 10 Mycobacterium due to the color contrast. In particular, when the MTB was grown, the size of the colonies was larger than with other media, so visualization was easy.

• Korean Journal of Clinical Laboratory Science
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• v.43 no.3
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• pp.120-123
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• 2011

A total of 30 water samples were collected from 30 different public baths in Seoul, Korea. Contamination of public bath water by waterborne pathogens can cause disease outbreaks and contribute to increase background rates of disease. Pathogens in water was filtered by nitrocellulose membrane with $0.45{\mu}m$ pore size. The membrane filters were analyzed by both cultivation and polymerase chain reaction (PCR) amplification of partial 16S rRNA gene. Various microorganisms including 4 Escherichia coli/Shigella spp. 1 Salmonella spp. 3 Pseudomonas aeruginosa and 2 Mycobacterium spp. were identified by reverse blot hybridization assay (REBA). PCR-REBA was able to identify many bacterial genera in one assay. Our results suggest that appropriate hygiene practice and continuous monitoring is needed for reducing health risk associated with public bath houses.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1992.05a
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• pp.22-22
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• 1992

국내의 토양에서 Streptomyces spp.를 분리하였고 이 중 항균활성이 우수한 균주들을 선별하였다. 이들 균주들을 적절한 조건에서 배양하여 얻은 배양액을 이온 교한 수지와 흡착 크로마토그래피를 행하여 항생물질들을 분리하였다. 이들 항생물질들은 Bacillus subtilis, Pyricularia oryzae, Alternaria mail, Candida albicans, Mycobacterium smegmatis, Pseudomonas aeruginosa , 및 Escherichia coli에 강한 항균활성을 나타내었다. 각 항생물질들을 산으로 가수분해하고 흡착관 크로마토그래피로 가수분해 생성물을 분리하였다. 분리한 항생물질과 가수분해 생성물들의 $^1$IH-NMR, $13^{C}$ -NMR, FAB-Mass 스펙트럼을 분석하여 구조를 연구하였다.