• 제목/요약/키워드: M. bovis

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Studies on Enzyme-linked Immunosorbent Assay(ELISA) for Detection of Antibody to Mycobacterium bovis in Serum and Milk (Enzyme-linked Immunosorbent Assay(ELISA)를 이용한 혈청 및 원유 중의 Mycobacterium bovis 항체 검출에 관한 연구)

  • 심항섭;국정희;박병옥;김성열;박유순
    • Korean Journal of Veterinary Service
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    • v.20 no.2
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    • pp.133-142
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    • 1997
  • In order to supplement a diagnostic method for detection of infectious cattle to bovine tuberculosis, performed ELISA for detection of antibody to if bovis in serum and milk. The diagnostic efficacy of the established ELISA was compared with test of the tuberculin skin test for bovine tuberculosis. The positive corresponding rate of serum ELISA and tuberculin skin test showed 84.3%, milk ELISA and tuberculin skin test showed 75.0%, milk ELISA and serum ELISA showed 75.0% respectively. Comparison of the serum and milk to tuberculin antibody concentration in tuberculin positive cattle, the milk contained 1/100-1/150 concentration compared serum tuberculin concentration. The established ELISA was considered efficient for detection of antibodies to M bovis in serum and milk.

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Detection of Mycobacterium bovis in the lymph node of tuberculin positive cattle by guanidium isothiocyanate/silica DNA extraction and polymerase chain reaction

  • Cho, Yun-Sang;Jung, Suk-Chan;Yoo, Han-Sang;Kim, Jong-Man
    • 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.

Rapid identification of bacterial pathogens related with bovine respitatory diseases by using PCR (PCR을 이용한 소 세균성 호흡기질병 원인체 신속동정)

  • Jung, Byeong-yeal
    • Korean Journal of Veterinary Research
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    • v.44 no.3
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    • pp.399-405
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    • 2004
  • Haemophilus somnus, Mycoplasma bovis and Pasteurella multocida were responsible for respiratory diseases in bovine. Methods for identifying these bacteria had poor sensitivity and specificity. In this paper, PCR assays were applied for rapid identification of H. somnus, M. bovis, P. multocida B:2 and P. multocida capsular types. The specific PCR products were amplified from H. somnus, but not from other bacteria. Ten-fold diluted H. somnus were mixed with P. multocida and then the mixed cultures were inoculated on agar plates. After incubation, PCR was performed with harvest from agar plates and could detect as few as 3.4 CFU/ml of H. somnus. The primers MboF and MboR produced an amplification product unique to M. bovis and sensitivity of PCR was as low as 100 pg of DNA. Only serotype B:2 of P. multocida, the causal agent of haemorrhagic septicemia in bovine, was specifically amplified in PCR among the 16 reference serotypes. The multiplex capsular PCR typing for P. multocida was produced the P. multocida specific product as well as the capsular serogroup-specific product. The present PCR assays should be useful for the rapid identification of bacterial pathogens from bovine respiratory diseases.

Clinical Bacteriological and Observations on Infectious Bovine Keratoconjunctivitis (IBK) in an Integrated Dairy Farm (대단위(大單位) 낙농목장(酪農牧場)에서의 소전염성결막각막염(傳染性結膜角膜炎)(IBK)의 임상적(臨床的) 및 세균학적(細菌學的) 관찰(觀察))

  • Seok, Ho-bong
    • Korean Journal of Veterinary Research
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    • v.25 no.1
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    • pp.53-59
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    • 1985
  • A group of 80 Holstein calves, many with clinical signs of infectious bovine keratoconjunctivitis in summer seasons, was assembled for bacteriologic and ophthalmologic studies at an integrated dairy farm. Observations were started from 1980 to 1984 but bacteriological study was only carried out from thor spring in 1981 and continued during the Autumn in 1981. Corneal lesions and drugs sensitivity were also observed and the results obtained were as follows: 1. Occurrences of IBK-affected cattle during the 5 years were predominent to calves in summer. 2. Among 142 eyes from 80 cattle, 53 isolates of Moraxella bovis(37.3%) were isolated from 37 cattle(46.3%). The prevalence of the infection for clinical eyes were higher than nonclinical eyes calves. 3. Miscellaneous organisms were found from age groups of 5 months to 9 months old. It were 8 hemolytic Neisseria spp., 3 Fungi, Mycoplasma sp. and 1 Pseudomonas sp., but 9 samples were unknown. 4. Prevalence of infection with M. bovis in bilateral clinical eyes were higher than unilateral eyes. 5. Incidence of corneal lesion was predominent in early stage as a watery tears(21.0%) and late stage as a leukomas of corneal opacity(22.8%). 6. The number of isolations of M. bovis and incidence of IBK varied from year to year; Higher incidence of IBK clinical signs were showed in calves from 5 months to 10 months old and the number of isolations of M. bovis was declined in order 8 months(100.0%), 4 months(75.0%), 5 months(66.6%) and 8 months(66.6%) old calves. 7. Chloramphenicol, Nitrofurantoin and Cephalothin were highly sensitive against M. bovis isolates and then Tetracycline, Neomycin, Erythromycin and Kanamycin were intermediate but low sensitive to Streptomycin, Colistin and Penicillin.

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Rapid diagnosis of bovine tuberculosis in slaughter cattle using PCR (PCR 기법을 이용한 도축 소의 결핵병 신속진단)

  • Koh, Ba-Ra-Da;Kim, Hyun-Joong;Park, Duk-Woong;Park, Seong-Do;Kim, Jae-Ik;Park, Jong-Tae;Kim, Yong-Hwan
    • Korean Journal of Veterinary Service
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    • v.30 no.3
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    • pp.393-406
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    • 2007
  • Bovine tuberculosis is an important zoonosis worldwide. Mycobacterium bovis, the causative agent of this disease in cattle, is also a pathogen for humans and several economically important animals. The cases of tuberculosis are reported in two cow found at slaughter house located in Gwangju city. Histopathologically, in the lymph nodes, granulomas consisted of large areas of necrosis surrounded by variable thick bands of cellular infiltrate containing macrophages, Langhans-type multinucleated giant cells and lymphocytes. Lesions in the lung followed the same developmental pattern as did lesions in the lymph nodes with some exceptions. With the acid-fast staining, numerous mycobacteria were revealed in the lung and lymph nodes. M bovis was confirmed as a causative agent in these cattle using bacterial isolation and PCR and restriction fragment length polymorphism method based on a unique 12.7 kb fragment insertion sequence from the Mycobacterium tuberculosis genome and the pncA polymorphism, The insertion element IS6110 and IS1081 were present M bovis isolated from lungs and lymph nodes of cattle using PCR assay. These cases are interesting and important in public health aspect that M bovis-infected cattle were found during a routine post-mortem inspection at slaughter house.

Studies on inoculation test of experimental animal and isolation and identificaton of Mycobacterium in tubercle and Iymph node of tuberculin(PPD) test positive dairy cattle. (Tuberculin(PPD)양성 반응우에 나타난 결핵 결절 및 림프절의 시험동물 접종 및 균분리 동정에 관한 연구)

  • Sung, Moung-Sook;Kim, Sin;Kim, Sang-Youn;Son, Jae-Kweon
    • Korean Journal of Veterinary Service
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    • v.20 no.2
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    • pp.205-215
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    • 1997
  • In this study, homogenized mixture of tubercle and pulmonary lymph node showed up tuberculin(PPD) positive dairy cattle was inoculated in experimental animal, and was cultured on medium(Lowenstein Jensen medium, 3% Ogawa). The results obtained through the survey were sumerized as follows ; 1. In experimental animal, goat and rabbits were decreased body weight(25∼28%) and died in 90 days with severe pathogenicity. Rats are slight pathogenicity. 2. Goat, rabbits and rats showed up severe lesions In lung, rabbits was also lesions other organs (kidney, appendix, ileocecum, and I lymph node). 3. Mycobacterium was grown between 5 weeks and 8 weeks on Lowenstein Jensen medium and 3% Ogawa medium. 4. Biochemical test of Mycobacterium cultured on medium was that niacin, nitrate reduction, tween 80 hydrolysis and catalase test were negative, but that urease test was positive. Therefore it was Mycobacterium bovis (M. bovis).

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Studies on the diagnosis of purified protein derivatives (PPD) tuberculin intradermal tuberculin test and ELISA to antibodies of Mycobacterium bovis (Purified protein derivatives(PPD) tuberculin 피내반응검사와 ELISA 항체가를 이용한 Mycobacterium bovis 진단에 관한 연구)

  • Chu, Keum-Suk;Cho, Bum-Jun;Cho, Young-Suk;Kang, Mi-Seon;Oh, Jin-Sik;Lee, Jeong-Won
    • Korean Journal of Veterinary Service
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    • v.32 no.2
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    • pp.125-129
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    • 2009
  • SBovine tuberculosis is a chronic bacterial disease of animals and humans caused by Mycobacterium bovis. Besides the classical intradermal tuberculin test, a number of blood and serum tests have been used. The purpose of this study was to establish seroprevalence of M. bovis. The sera were screened using the ELISA technique. A total seroprevalence of 65.8% in positive cattle, suspect 36.0%, negative 5.9% in TB-infected herds by PPD and dairy cattle is 3.0%, Hanwoo is 1.6% in TB-free herds. The deer of seroprevalence is 55.0% in TB-infected herd and 7.7% in TB-free herds.

A Case of Disseminated Mycobacterium bovis Infection after BCG Vaccination (Tokyo strain) in an Apparently Immunocompetent Infant (면역기능이 정상인 영아에서 동경주 BCG 백신 접종 후 발생한 파종성 Mycobacterium bovis 감염 1예)

  • Paik, Ji Yeun;Choi, Jae Hong;Kim, Min Kyung;Choi, Eun Hwa;Lee, Hoan Jong;Park, Kyoung Un
    • Pediatric Infection and Vaccine
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    • v.18 no.1
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    • pp.91-96
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    • 2011
  • Bacille Calmette-Gu$\acute{e}$rin (BCG) vaccine is a live attenuated vaccine derived from Mycobacterium bovis. Frequent complications after BCG vaccination are localized ulcer formation and regional lymphadenitis, but there could be rarely severe systemic reactions to BCG vaccine such as osteomyelitis and disseminated BCG infection. Although disseminated BCG infection can be complicated in infants with underlying immunodeficiency after BCG vaccination, it is very unlikely to develop in immunocompetent infants or children. We report a 13-month-old infant who presented with fever, skin nodules, and multiple enlarged lymph nodes 5 months following BCG vaccination. She was diagnosed with disseminated BCG infection by PCRconfirmed M. bovis BCG infection at ${\geq}$2 anatomical sites beyond the region of vaccination. The patient showed no obvious evidence of immunodeficiency as judged on the basis of previous disease history, plasma immunoglobulin levels, B and T lymphocytes counts in peripheral blood, DHR (dihydrorhodamine 123 fluorescence) test and HIV test. She started antituberculous treatment with isoniazid and rifampin, and now, apparently her symptoms have been improved.

Cloning and Expression of Mycobacterium bovis Secreted Protein MPB83 in Escherichia coli

  • Xiu-Yun, Jiang;Wang, Chun-Feng;Wang, Chun-Fang;Zhang, Peng-Ju;He, Zhao-Yang
    • BMB Reports
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    • v.39 no.1
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    • pp.22-25
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    • 2006
  • The gene encoding MPB83 from Mycobacterium bovis Vallee111 chromosomal DNA was amplified by using polymerase chain reaction (PCR) technique, and the PCR product was approximately 600bp DNA segment. Using T-A cloning technique, the PCR product was cloned into pGEM-T vector and the cloning plasmid pGEM-T-83 was constructed successfully. pGEM-T-83 and pET28a(+) were digested by BamHI and EcoRI double enzymes. The purified MPB83 gene was subcloned into the expression vector pET28a(+), and the prokaryotic expression vector pET28a-83 was constructed. Plasmid containing pET28a-83 was transformed into competence Escherichia coli BL21 (DE3). The bacterium was induced by isopropyl-$\beta$-D-thiogalactopyranoside (IPTG) and its lysates were loaded directly onto sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), approximately 26 kDa exogenous protein was observed on the SDS-PAGE. The protein was analyzed using Western-blotting. The results indicated that the protein was of antigenic activity of M. bovis. The results were expected to lay foundation for further studies on the subunit vaccine and DNA vaccine of MPB83 gene in their prevention against bovine tuberculosis.

The Scavenging Effect on Nitric Oxide in Ursi Fel ${\cdot}$ Bovis Calculus Herbal-acupuncture Solution (웅담합우황(熊膽合牛黃) 약침액(藥鍼液)의 Nitric Oxide에 대한 억제효과(抑制效果))

  • Woo, Chang-Hoon;Lee, Kyung-Min;Lee, Bong-Hyo;Lim, Seong-Chul;Jung, Tae-Young;Seo, Jung-Chul
    • Journal of Acupuncture Research
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    • v.24 no.2
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    • pp.221-229
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    • 2007
  • Oxidative damage to DNA, proteins, lipids and other molecules may contribute to the development of neurodegenerative disease. This study was designed to find out whether Ursi Fel ${\cdot}$ Bovis Calculus Herbal-Acupuncture Solution(UBHAS) can scavenge nitric oxide(NO) or not. NO concentration was estimated after treatment with Vit. C or 1, 10, $100{\mu}g/m{\ell}$ of UBHAS. There was no significant scavenging effect of UBHAS on NO in 1, $100{\mu}g/m{\ell}$ group. while significant scavenging in $l0{\mu}g/m{\ell}$ group after 2hrs. UBHAS showed significant scavenging effects on NO in 1, 10, $100{\mu}g/m{\ell}$ group after 6hrs. There was a significant scavenging effect in 10, $100{\mu}g/m{\ell}$ group. even though no significant in $1{\mu}g/m{\ell}$ group after 12, 24 hrs. This study showed that UBHAS might be used for aging related disease. These results suggest that UBHAS has scavenging effect on NO. however, further studies are required to investigate the antioxidative effects of it.

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