• Title/Summary/Keyword: M. bovis

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Transcriptional and Mycolic Acid Profiling in Mycobacterium bovis BCG In Vitro Show an Effect for c-di-GMP and Overlap between Dormancy and Biofilms

  • Cruz, Miguel A. De la;Ares, Miguel A.;Rodriguez-Valverde, Diana;Vallejo-Cardona, Alba Adriana;Flores-Valdez, Mario Alberto;Nunez, Iris Denisse Cota;Aceves-Sanchez, Michel de Jesus;Lira-Chavez, Jonahtan;Rodriguez-Campos, Jacobo;Bravo-Madrigal, Jorge
    • Journal of Microbiology and Biotechnology
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    • v.30 no.6
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    • pp.811-821
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    • 2020
  • Mycobacterium tuberculosis produces mycolic acids which are relevant for persistence, recalcitrance to antibiotics and defiance to host immunity. c-di-GMP is a second messenger involved in transition from planktonic cells to biofilms, whose levels are controlled by diguanylate cyclases (DGC) and phosphodiesterases (PDE). The transcriptional regulator dosR, is involved in response to low oxygen, a condition likely happening to a subset of cells within biofilms. Here, we found that in M. bovis BCG, expression of both BCG1416c and BCG1419c genes, which code for a DGC and a PDE, respectively, decreased in both stationary phase and during biofilm production. The kasA, kasB, and fas genes, which are involved in mycolic acid biosynthesis, were induced in biofilm cultures, as was dosR, therefore suggesting an inverse correlation in their expression compared with that of genes involved in c-di-GMP metabolism. The relative abundance within trehalose dimycolate (TDM) of α-mycolates decreased during biofilm maturation, with methoxy mycolates increasing over time, and keto species remaining practically stable. Moreover, addition of synthetic c-di-GMP to mid-log phase BCG cultures reduced methoxy mycolates, increased keto species and practically did not affect α-mycolates, showing a differential effect of c-di-GMP on keto- and methoxy-mycolic acid metabolism.

Bovine Tuberculosis of Korean Native Cattle in an Abattoir (도축장 내의 한우에서 발견된 소 결핵 보고)

  • Jang, Seong-Jun;Do, Sun-Hee;Ki, Mi-Ran;Hong, Il-Hwa;Park, Jin-Kyu;Ji, Ae-Ri;Jeong, Kyu-Shik
    • Journal of Life Science
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    • v.19 no.12
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    • pp.1847-1850
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    • 2009
  • Bovine tuberculosis is generally detected postmortem because it is a chronic debilitating disease. Since tuberculosis is mainly found in the lungs, clinical signs including coughing, nasal discharge, and difficulty breathing can occur in severe instances. In the present study, specimens were collected from the heart, liver, kidney, lung, pleural cavities, lymph nodes and intestines of carcasses found in an abattoir. According to post-mortem examination and inspection of carcasses, tuberculosis lesions were varied in appearance and size. Tubercles of a white cream color were disseminated throughout the pleural cavity including the lymph nodes, lungs and mesentery containing pus. Gram and Ziehl-Neelsen's acid-fast staining for the lung and lymph nodes revealed a highly positive histochemical reaction. The acid-fast organisms were observed histologically in the lesions under a microscope. This report demonstrated the histopathology of bovine tuberculosis based on the histological findings of Mycobacterium bovis, which is a suspected causative agent.

Studies on the Tuberculin Production - (3) Studies on the Change of Tuberculin Reactivity when Repeated Tuberculin Injection (Tuberculin에 관한 연구(硏究) - (3) Tuberculin 반응(反應)을 반복(反復)할 때 일어나는 변화(變化)에 관한 관찰(觀察))

  • Kim, Jung-Kyu
    • The Journal of the Korean Society for Microbiology
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    • v.6 no.1
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    • pp.41-60
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    • 1971
  • The author prepared a tuberculin sample using synthetic medium for the test material in this experiment. For the test animals, guinea pigs, healthy calf sensitized with heat killed tubercle bacilli, naturally infected dairy cattle, goat, and calf were used. A reduced reactivity to tuberculin was observed when the second tuberculin test was done within 28 days after the first. The results obtained are summarized as follows. 1. The experimental HCSM tuberculin was shown the similar activities as the control products of standard tuberculin. 2. The potency of the experimental tuberculin preserved at $5^{\circ}C$ in refrigerator maintained the activity for three years. 3. By the intradermal test, the experimental tuberculin was proved to be specific to detect tuberculous dairy cattles and goats. 4. The appearance of skin reaction to tuberculin in naturally infected dairy cattles, goat and beef cattles was soft and diffuse swellings or circumscribed swellings accompanied by induration. 5. In the case of observing tuberculous dairy cattles, goats and beef cattles, as the criterion of the reaction to the caudal fold test, results were taken as positive when findings are either hard and circumscribed or soft and diffuse swellings of 3 mm at least in size. 6. The acid fast organisms isolated from tuberculin positive animals were identified mostly as M. tuberculosis var. bovis. 7. When the tuberculin test was repeated in the tuberculin positive animals, the reactivity decreased rapidly, lasting 4 weeks at least.

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Supraclavicular BCG Lymphadenitis Noted at 21 Months after BCG Vaccination Confirmed by a Molecular Method (분자생물학적 방법으로 확진한 접종 21개월 후에 발생한 BCG 쇄골상부 림프절염 1례)

  • Lee, Min Hyun;Chae, Moon-Hee;Park, Kyoung Un;Cho, Hye-Kyung
    • Pediatric Infection and Vaccine
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    • v.21 no.2
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    • pp.139-143
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    • 2014
  • Bacille Calmette-$Gu\acute{e}rin$ (BCG) lymphadenitis is the most common complication of BCG vaccination. It commonly occurs in infants aged <6 months involving ipsilateral axillary lymph nodes. We described BCG lymphadenitis in a 22-month-old boy presenting swelling of left supraclavicular lymph node that was confirmed by real-time polymerase chain reaction (PCR) and the multiplex PCR targeting the region of difference (RD).

Comparative Proteomic Analysis of Virulent Korean Mycobacterium tuberculosis K-strain with Other Mycobacteria Strain Following Infection of U-937 Macrophage

  • Ryoo, Sung-Weon;Park, Young-Kil;Park, Sue-Nie;Shim, Young-Soo;Liew, Hyun-Jeong;Kang, Seong-Man;Bai, Gill-Han
    • Journal of Microbiology
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    • v.45 no.3
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    • pp.268-271
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    • 2007
  • In Korea, the Mycobacterium tuberculosis K-strain is the most prevalent clinical isolates and belongs to the Beijing family. In this study, we conducted comparative porteomics of expressed proteins of clinical isolates of the K-strain with H37Rv, H37Ra as well as the vaccine strain of Mycobacterium bovis BCG following phagocytosis by the human monocytic cell line U-937. Proteins were analyzed by 2-D PAGE and MALDI-TOF-MS. Two proteins, Mb1363 (probable glycogen phosphorylase GlgP) and MT2656 (Haloalkane dehalogenase LinB) were most abundant after phagocytosis of M. tuberculosis K-strain. This approach provides a method to determine specific proteins that may have critical roles in tuberculosis pathogenesis.

Analysis of partial cDNA sequence from Theileria sergenti

  • Park, Jin-ho;Chae, Joon-seok;Kim, Dae-hyuk;Jang, Yong-suk;Kwon, Oh-deog;Lee, Joo-mook
    • Korean Journal of Veterinary Research
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    • v.39 no.4
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    • pp.797-805
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    • 1999
  • T sergenti cDNA library were constructed to get a more broad information about the structural, functional or antigenic properties of the proteins, and analyzes for their partial cDNA sequences and expression sequences tags(ESTg). The mRNA were purified from T sergenti isolates to identify the information of antigen gene, then first and second strand cDNA was synthesized. EcoR I adaptor ligation and Xho I enzyme restriction were used to the synthesized cDNA, and ligated into a Uni-ZAP XR vector. T sergenti cDNA library was constructed with packaging and amplification in vitro. Antibody screening was performed with constructed T sergenti cDNA library using antisera against T sergenti. Among those clones, eight phagemids were rescued from the recombinant in vivo excision with f1 helper phage. Using the analysis of endonuclease restriction and PCR, the recombinant cDNA were proved having a 0.5-3.0kb of inserts. The eight of partial cDNA clones' sequences were obtained and examined for their homology using BLASTN and BLASTX. The eight of sequenced clones were classified into three groups according to the basis of database searches. A total 3,045bp of partial cDNA sequence were determined from six clones. The putatively identified clones contain a cytochrome c gene, a heat shock protein gene, a cyclophilin gene, and a ribosomal protein gene. The unidentified clones have a homology to ATP-binding protein(mtrA) gene of S argillaceus, DNA-binding protein(DBP) gene of Pseudorabies virus 85kDa merozoite protein gene of B bovis, mRNA spm1 protein of T annulata and glycine-rich RNA-binding protein mRNA of O sativa etc.

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Pure Isolation and Identification of Babesia ovata by Morphological Characteristics and Complement Fixation Test in Korea (Babesia ovata의 순수분리와 형태학적 및 혈청학적 동정)

  • Suh, Myung-deuk
    • Korean Journal of Veterinary Research
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    • v.27 no.2
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    • pp.307-316
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    • 1987
  • This study was conducted to identify Babesia sp. strain isolated from the imported beef cattle, Aberdeen-angus heifers, in J farm of Jangsu Prefecture, Cheon-buk Province of Korea, morphologically, etiologically and serologically. Babesia sp. strain was purely isolated through the serial blood passages of three generations against splenectomized calves and one generation of blood passage against non-splenectomized calf(intact calf) by inhibiting the appearance of Theileria sergenti in the blood stream by means of injection of 20% oil pamaquine intramuscularly. In the splenectomized calves, the parasite multiplied markedly in blood stream soon after inoculation and parasitaemia was much severe. An elevated body temperature, anorexia, severe anaemia and icterus were observed clinically. Of three splenectomized calves, two were affected with haemoglobinuria and died. But in the non-splenectomized calf the clinical signs and parasitaemia were very mild. The means of the incubation period, the highest parasitaemia, the highest body temperature and the lowest PCV were 3.5 days, 41.1%, ${42^{\circ}C}$ and 9%, respectively, in the splenectomized calves. In calf erythrocytes Babesia sp. protozoa were polymorphic showing the round, oval, ameboid, piriform and paired piriform etc. The sizes of protozoa were $2.51{\sim}3.91{\times}1.32{\sim}2.19{{\mu}m}$ ($3.20{\times}1.60{\mu}m$). Serologically the isolated Babesia sp. were compared with other parasites, B. ovata, B. bigemina, B. bovis, T. sergenti, A. marginale and A. centrale by using the complement fixation(CF) test. As a result, the antibody titer between the homologous species were high. It was two tubes or more in the CF test. From the results obtained above the pure isolated Babesia species was identical with Babesia ovata Minami and Ishihara, 1980.

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Expression of the 38 kDa Protein of Mycobacterium tuberculosis in M. bovis BCG and Use in the Serodiagnosis of Tuberculosis

  • Cho, Sang-Nae;Kim, Hee-Jin;Lee, Hye-Young;Kim, Seung-Chul;Kim, Joo-Deuk
    • The Journal of the Korean Society for Microbiology
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    • v.34 no.6
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    • pp.555-559
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    • 1999
  • The 38 kDa protein of Mycobacterium tuberculosis, which was known previously as antigen 5, has been extensively used in the serodiagnosis of tuberculosis. In an attempt to develop and evaluate a serodiagnostic test using the antigen, we expressed the 38 kDa protein in BCG and its seroreactivity was compared to that expressed in Escherichia coli. The coding region of the 38 kDa protein was amplified by PCR, and the gene was cloned into a Mycobacterium-E. coli shuttle expression vector pYMC-his and pQE30 expression vector and expressed in BCG and E. coli, respectively. Both recombinant 38 kDa proteins showed strong seroreactivity against pooled serum from tuberculosis patients. There was no significant difference in seroreactivity between the two recombinant antigens in sera from the far advanced tuberculosis patients. However, of 25 tuberculosis patients graded as "minimal" by chest X-ray, 5 (20.0%) were seropositive by r38 kDa expressed in E. coli, while 8 (32.0%) by that expressed in BCG. Likewise, higher seroreactivity by r38 kDa expressed in BCG was found in sera from the moderately advanced tuberculosis. This study thus indicates that the recombinant 38 kDa expressed in BCG is more effective than that expressed in E. coli in detecting antibodies to the native 38 kDa protein of M. tuberculosis in sera from minimally affected tuberculosis patients.

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Evaluation of microbiological, cellular and risk factors associated with subclinical mastitis in female buffaloes

  • de Oliveira Moura, Emmanuella;do Nascimento Rangel, Adriano Henrique;de Melo, Maria Celeste Nunes;Borba, Luiz Henrique Fernandes;de Lima, Dorgival Morais Junior;Novaes, Luciano Patto;Urbano, Stela Antas;de Andrade Neto, Julio Cesar
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.9
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    • pp.1340-1349
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    • 2017
  • Objective: This study aimed to evaluate the microbiological and cellular milk profile for the diagnosis of subclinical mastitis in female buffaloes and to assess risk factors for predisposition of the disease. Methods: Analyses were carried out by standard plate count (SPC), identification of species and antibiotic resistance, somatic cell count (SCC), electrical electrical conductivity of milk (ECM), and lactoferrin content in milk. Teat cups were swabbed to evaluate risk factors, observing hyperkeratosis, milking vacuum pressure and cleanliness of the site. Hence, 30 female buffaloes were randomly selected (15 from a group in early lactation and 15 in late lactation). Results: The most common bacteria in the microbiological examination were Staphylococcus spp., Streptococcus spp. and Corynebacterium sp. In the antibiotic sensitivity test, 10 (58.82%) of the 17 antibiotics tested were sensitive to all isolates, and resistant bacteria were Streptococcus uberis, Streptococcus dysgalactiae, Streptococcus haemolyticus, and Escherichia coli. It was observed that positive samples in the microbiological examination showed total bacterial count between $9.10{\times}10^3$ to $6.94{\times}10^6$ colony forming units/mL, SCC between 42,000 to 4,320,000 cells/mL and ECM ranging from 1.85 to 7.40 mS/cm. It was also found that the teat cups had high microbial counts indicating poor hygiene, and even faults in the cleanliness of the animals' waiting room were observed. It is concluded that values of SCC above 537,000 cells/mL and ECM above 3.0 mS/mL are indications of mammary gland infection for this herd; however, the association of these values with a microbiological analysis is necessary to more accurately evaluate the health status of mammary glands with subclinical mastitis. Conclusion: Through phenotypic characterization of bacteria involved in the samples, the genera Staphylococcus spp., Streptococcus spp., and Corynebacterimum bovis were the most prevalent in this study. Faults in environment and equipment hygienization are factors that are directly associated with mastitis.

Prevalence of infectious agents in cattle reared in Ulleung island (울릉도 소의 전염성 병원체 감염률 조사)

  • Seo, Min-Goo;Do, Jae-Cheul;Ouh, In-Ohk;Coh, Min-Hee;Kim, Joong-Kew;Kim, Young-Hoan;Park, No-Chan;Kwak, Dong-Mi
    • Korean Journal of Veterinary Service
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    • v.34 no.4
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    • pp.303-311
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    • 2011
  • Prevalence of infectious agents, including Brucella abortus (BA), Mycobacterium bovis (MB), bovine leukemia virus (BLV), M. avium subsp. paratuberculosis (MP), Neospora caninum (NC) and Toxoplasma gondii (TG), was investigated in all the cattle raised in Ulleung island during 2007~2010. For BA, the prevalences in head and farm were 8.1% (44/545) and 5.5% (4/73) in 2007, all negative in 2008~2009, and 0.5% (4/774) and 1.7% (1/58) in 2010, respectively. For MB, no sample was positive by PPD or ELISA in 2007~2010. For BLV and MP, no sample was positive by ELISA in 2007~2009. For NC, seroprevalences in head and farm were 0.2% (1/545) and 1.4% (1/73), respectively, in 2007 and all negative in 2008~2009. For TG, seroprevalences in head and farm were 17.6% (97/552) and 54.8% (34/62) by ELISA in 2009. By regions, the seroprevalences of TG in Ulleung-eup, Seo-myeon and Buk-myeon were 26.0%, 9.8% and 16.7%, respectively, which had significant differences (P<0.0001). Tiger cattle were more resistant to TG infection than Hanwoo. The seroprevalence of TG in summer was higher than in autumn. The seroprevalence of TG in cows was higher than in oxen. The seroprevalence of TG in cattle was increased with age. In conclusion, this study indicates that the prevalences of six infectious diseases, except for TG which are widely spread, are relatively low in cattle reared in Ulleung island.