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Sample records for bovis bacillus calmette-guerin

  1. Prosthetic Joint Infection due to Mycobacterium bovis after Intravesical Instillation of Bacillus Calmette-Guerin (BCG

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    Eric Gomez

    2009-01-01

    Full Text Available Intravesical instillation of Bacillus Calmette-Guerin (BCG is a treatment to prevent recurrence of superficial urothelial bladder carcinoma. Complications after bladder instillation of BCG have been reported including locally invasive and systemic infections due to dissemination of Mycobacterium bovis from the bladder. We present an uncommon case and literature review of prosthetic joint infection due to M. bovis after intravesical BCG treatment of bladder cancer.

  2. Mycobacterium bovis-Bacillus Calmette-Guerin and asthma

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    SHEN Hua-hao; ZHANG Gen-sheng; WANG Ping-li

    2005-01-01

    @@ In recent decades, the prevalence of asthma over the world has risen steadily especially in high-income countries.1 The underlying mechanisms for this phenomenon remain largely unknow, but environmental factors are thought to play an important role. Some epidemiology studies2-8 suggested that the increase in asthma inversely correlated with a steady decline exposition to some human diseases, such as tuberculosis and influenza. One of the general features of these infectious agents is that they induce characteristic Th1 type immune response which suppresses the Th2 response predominate in individuals suffering from asthma. Bacille Calmette-Guerin(BCG) is a strong inducer of Th1 type immune response. This review will focus on recent findings related to the interaction between BCG and asthma, and the main mechanisms of BCG in modulating asthma development.

  3. Mutation in alkylhydroperoxidase D gene dramatically decreases persistence of Mycobacterium bovis bacillus calmette-guerin in infected macrophage

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    Farivar Taghi

    2008-07-01

    Full Text Available Background and Objectives: Mycobacterium tuberculosis is the leading cause of death from a single bacterial species in the world and is subjected to a highly oxidative environment in its host macrophage and consequently has evolved protective mechanisms against reactive oxygen and nitrogen intermediates. Alkyl hydroperoxidase D (AhpD is a molecule from these mycobacterial defense systems that has a dual function. It not only works with Alkyl hydroperoxidase C (AhpC in mycobacterial defense system against oxidative stress but also has a role in oxidation/reduction of succinyltransferase B (SucB, dihydrolipoamide dehydrogenase (LPD and AhpC. The present study was undertaken to find out the effects of inactivation of ahpD gene in the intra-macrophage persistence of resulted BCG mutant. Materials and Methods: We did allelic exchange mutagenesis in Mycobacterium bovis BCG and evaluate the effects of this mutagenesis in intracellular persistence of wild type BCG strains and ahpD mutant ones by comparing colony forming units (CFU in infected macrophage. Results: Our findings showed that after producing allelic exchange mutagenesis in ahpD gene of M.bovis BCG a sever decrease in the CFU′s of ahpD mutant BCG strains has been observed and intracellular persistence of ahpD mutant BCG strains decreased significantly. Conclusion: Mutagenesis in ahpD gene will cause significant decrease in intracellular survival of ahpD mutant strains than wild type M.bovis BCG strains and could leads to an inefficiency in pyruvate dehydrogenase pathway and could also impair impairs mycobacterial defense system against oxidative and nitrosative stress.

  4. Disseminated Bacillus Calmette Guerin Disease in a Twin Infant with Severe Combined Immunodeficiency Disease

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    Mittal, Hema; Faridi, MMA; Kumar, Pankaj; Aggarwal, Anju

    2014-01-01

    Fatal-disseminated Bacillus Calmette Guerin (BCG) disease is well known in infants with severe combined immunodeficiency after BCG vaccination. We report a 7 month male infant delivered as a product of in vitro fertilization and twin gestation that presented with fever, cough and multiple nodular skin lesions. A biopsy of skin lesions revealed the presence of acid fast bacilli. Mycobacterium bovis infection was confirmed by polymerase chain reaction (PCR) and molecular studies. Immunological profile confirmed the diagnosis of severe combined immunodeficiency. Only few reports of similar case exist in the literature. PMID:25191057

  5. Disseminated bacillus calmette guerin disease in a twin infant with severe combined immunodeficiency disease

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    Hema Mittal

    2014-01-01

    Full Text Available Fatal-disseminated Bacillus Calmette Guerin (BCG disease is well known in infants with severe combined immunodeficiency after BCG vaccination. We report a 7 month male infant delivered as a product of in vitro fertilization and twin gestation that presented with fever, cough and multiple nodular skin lesions. A biopsy of skin lesions revealed the presence of acid fast bacilli. Mycobacterium bovis infection was confirmed by polymerase chain reaction (PCR and molecular studies. Immunological profile confirmed the diagnosis of severe combined immunodeficiency. Only few reports of similar case exist in the literature.

  6. Combined thermo-chemotherapy for recurrent bladder cancer after bacillus Calmette-Guerin.

    NARCIS (Netherlands)

    Nativ, O.; Witjes, J.A.; Hendricksen, K.; Cohen, M.; Kedar, D.; Sidi, A.; Colombo, R.; Leibovitch, I.

    2009-01-01

    PURPOSE: Despite an initial adequate response many patients with nonmuscle invasive urothelial cell carcinoma of the bladder eventually have recurrence after intravesical bacillus Calmette-Guerin treatments. We evaluated the efficacy of combined bladder wall hyperthermia and intravesical mitomycin C

  7. Oral vaccination of guinea pigs with a Mycobacterium bovis bacillus Calmette-Guerin vaccine in a lipid matrix protects against aerosol infection with virulent M. bovis.

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    Clark, Simon; Cross, Martin L; Nadian, Allan; Vipond, Julia; Court, Pinar; Williams, Ann; Hewinson, R Glyn; Aldwell, Frank E; Chambers, Mark A

    2008-08-01

    Increased incidence of bovine tuberculosis (TB) in the United Kingdom caused by infection with Mycobacterium bovis is a cause of considerable economic loss to farmers and the government. The Eurasian badger (Meles meles) represents a wildlife source of recurrent M. bovis infections of cattle in the United Kingdom, and its vaccination against TB with M. bovis bacillus Calmette-Guérin (BCG) is an attractive disease control option. Delivery of BCG in oral bait holds the best prospect for vaccinating badgers over a wide geographical area. Using a guinea pig pulmonary challenge model, we evaluated the protective efficacy of candidate badger oral vaccines, based on broth-grown or ball-milled BCG, delivered either as aqueous suspensions or formulated in two lipids with differing fatty acid profiles (one being animal derived and the other being vegetable derived). Protection was determined in terms of increasing body weight after aerosol challenge with virulent M. bovis, reduced dissemination of M. bovis to the spleen, and, in the case of one oral formulation, restricted growth of M. bovis in the lungs. Only oral BCG formulated in lipid gave significant protection. These data point to the potential of the BCG-lipid formulation for further development as a tool for controlling tuberculosis in badgers.

  8. A novel recombinant Mycobacterium bovis bacillus Calmette-Guerin strain expressing human granulocyte macrophage colony-stimulating factor and Mycobacterium tuberculosis early secretory antigenic target 6 complex augments Th1 immunity

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    Xiaoling Yang; Lang Bao; Yihao Deng

    2011-01-01

    Since Mycobacterium bovis bacillus Calmette-Guerin strain (BCG) fails to protect adults from pulmonary tuberculosis (TB), there is an urgent need for developing a new vaccine. In this study, we constructed a novel recombinant BCG strain (rBCG) expressing human granulocyte macrophage colony-stimulating factor (GM-CSF) and the 6 kDa early secretory antigenic target (ESAT6) of Mycobacteriutn tuberculosis, named rBCG:GE (expressing GMCSFESAT6 complex), and evaluated the immunogenicity of the construct in BALB/c mice. Our results indicated that the rBCG:GE was able to induce higher titer of antibody than the conventional BCG, the rBCG:G (expressing GM-CSF)and the rBCG:E (expressing ESAT6). Moreover, the rBCG:GE also elicited a longer-lasting and stronger Thl cellular immune responses than the other groups, which was confirmed by the incremental proliferation of splenocytes, the increased percentages of CD4+ and CD8+ T cells of spleen, the elevated level of interferon-γ in splenocyte culture after tuberculin-purified protein derivative stimulation, and the increased concentration of GM-CSF in serum. The data presented here suggested the possibility that the recombinant BCG:GE might be a good vaccine candidate to TB.

  9. Treatment Options Available for Bacillus Calmette-Guerin Failure in Non-muscle-invasive Bladder Cancer

    NARCIS (Netherlands)

    Yates, D.R.; Brausi, M.A.; Catto, J.W.; Dalbagni, G.; Roupret, M.; Shariat, S.F.; Sylvester, R.J.; Witjes, J.A.; Zlotta, A.R.; Palou-Redorta, J.

    2012-01-01

    CONTEXT: Intravesical bacillus Calmette-Guerin (BCG) is a standard conservative treatment for patients with high-risk non-muscle-invasive bladder cancer (NMIBC). Many patients will experience recurrence or progression following BCG and are termed BCG failures. OBJECTIVE: To summarise the current tre

  10. Tuberculous Spondylitis following Intravesical Bacillus Calmette-Guerin for Bladder Cancer

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    Masashi Miyazaki

    2016-01-01

    Full Text Available We present a rare case of tuberculous spondylitis following intravesical Bacillus Calmette-Guerin (BCG therapy for bladder cancer. An 82-year-old man presented with low back pain. Past medical history revealed bladder cancer diagnosed and treated 16 months previously by intravesical BCG. Magnetic resonance imaging of the thoracic spine showed destruction of the T5 and T6 vertebrae and an epidural soft tissue mass with anterior dural sac compression. Due to the progression of vertebral destruction, posterior spinal segmental fusion was performed. Mycobacterium bovis (M. bovis was identified using multiplex polymerase chain reaction of surgical tissue specimens. The patient was started on an antituberculosis treatment regimen including isoniazid, rifampicin, and ethambutol. After surgery, his back pain resolved completely. At the latest examination, the patient was pain-free with no functional limitations or recurrent infection in clinical or imaging findings. Patients undergoing BCG therapy should be monitored for possible hematogenous spread of mycobacteria to the spine for months or even years after treatment.

  11. Effects of dietary glutamine supplementation on the body composition and protein status of early-weaned mice inoculated with Mycobacterium bovis Bacillus Calmette-Guerin.

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    Rogero, Marcelo Macedo; Borges, Maria Carolina; de Castro, Inar Alves; Pires, Ivanir S O; Borelli, Primavera; Tirapegui, Julio

    2011-09-01

    Glutamine, one of the most abundant amino acids found in maternal milk, favors protein anabolism. Early-weaned babies are deprived of this source of glutamine, in a period during which endogenous biosynthesis may be insufficient for tissue needs in states of metabolic stress, mainly during infections. The objective of this study was to verify the effects of dietary glutamine supplementation on the body composition and visceral protein status of early-weaned mice inoculated with Mycobacterium bovis Bacillus Calmette-Guérin (BCG). Mice were weaned early on their 14th day of life and seperated into two groups, one of which was fed a glutamine-free diet (n = 16) and the other a glutamine-supplemented diet (40 g/kg diet) (n = 16). At 21 days of age, some mice were intraperitoneally injected with BCG. Euthanasia was performed at the 28th day of age. BCG inoculation significantly reduced body weight (P glutamine supplementation resulted in a significant increase in serum IGF-1 (P = 0.019) and albumin (P = 0.025) concentration, muscle protein concentration (P = 0.035) and lipid content (P = 0.002) in the carcass. In conclusion, dietary glutamine supplementation had a positive influence on visceral protein status but did not affect body composition in early-weaned mice inoculated with BCG.

  12. Effects of Dietary Glutamine Supplementation on the Body Composition and Protein Status of Early-Weaned Mice Inoculated with Mycobacterium bovis Bacillus Calmette-Guerin

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    Ivanir S. O. Pires

    2011-08-01

    Full Text Available Glutamine, one of the most abundant amino acids found in maternal milk, favors protein anabolism. Early-weaned babies are deprived of this source of glutamine, in a period during which endogenous biosynthesis may be insufficient for tissue needs in states of metabolic stress, mainly during infections. The objective of this study was to verify the effects of dietary glutamine supplementation on the body composition and visceral protein status of early-weaned mice inoculated with Mycobacterium bovis Bacillus Calmette-Guérin (BCG. Mice were weaned early on their 14th day of life and seperated into two groups, one of which was fed a glutamine-free diet (n = 16 and the other a glutamine-supplemented diet (40 g/kg diet (n = 16. At 21 days of age, some mice were intraperitoneally injected with BCG. Euthanasia was performed at the 28th day of age. BCG inoculation significantly reduced body weight (P < 0.001, lean mass (P = 0.002, water (P = 0.006, protein (P = 0.007 and lipid content (P = 0.001 in the carcass. Dietary glutamine supplementation resulted in a significant increase in serum IGF-1 (P = 0.019 and albumin (P = 0.025 concentration, muscle protein concentration (P = 0.035 and lipid content (P = 0.002 in the carcass. In conclusion, dietary glutamine supplementation had a positive influence on visceral protein status but did not affect body composition in early-weaned mice inoculated with BCG.

  13. Effects of Dietary Glutamine Supplementation on the Body Composition and Protein Status of Early-Weaned Mice Inoculated with Mycobacterium bovis Bacillus Calmette-Guerin

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    Rogero, Marcelo Macedo; Borges, Maria Carolina; de Castro, Inar Alves; Pires, Ivanir S. O.; Borelli, Primavera; Tirapegui, Julio

    2011-01-01

    Glutamine, one of the most abundant amino acids found in maternal milk, favors protein anabolism. Early-weaned babies are deprived of this source of glutamine, in a period during which endogenous biosynthesis may be insufficient for tissue needs in states of metabolic stress, mainly during infections. The objective of this study was to verify the effects of dietary glutamine supplementation on the body composition and visceral protein status of early-weaned mice inoculated with Mycobacterium bovis Bacillus Calmette-Guérin (BCG). Mice were weaned early on their 14th day of life and seperated into two groups, one of which was fed a glutamine-free diet (n = 16) and the other a glutamine-supplemented diet (40 g/kg diet) (n = 16). At 21 days of age, some mice were intraperitoneally injected with BCG. Euthanasia was performed at the 28th day of age. BCG inoculation significantly reduced body weight (P < 0.001), lean mass (P = 0.002), water (P = 0.006), protein (P = 0.007) and lipid content (P = 0.001) in the carcass. Dietary glutamine supplementation resulted in a significant increase in serum IGF-1 (P = 0.019) and albumin (P = 0.025) concentration, muscle protein concentration (P = 0.035) and lipid content (P = 0.002) in the carcass. In conclusion, dietary glutamine supplementation had a positive influence on visceral protein status but did not affect body composition in early-weaned mice inoculated with BCG. PMID:22254124

  14. Improving Mycobacterium bovis bacillus Calmette-Guerin as a vaccine delivery vector for viral antigens by incorporation of glycolipid activators of NKT cells.

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    Manjunatha M Venkataswamy

    Full Text Available Recombinant Mycobacterium bovis bacillus Calmette-Guèrin (rBCG has been explored as a vector for vaccines against HIV because of its ability to induce long lasting humoral and cell mediated immune responses. To maximize the potential for rBCG vaccines to induce effective immunity against HIV, various strategies are being employed to improve its ability to prime CD8+ T cells, which play an important role in the control of HIV infections. In this study we adopted a previously described approach of incorporating glycolipids that activate CD1d-restricted natural killer T (NKT cells to enhance priming of CD8+ T cells by rBCG strains expressing an SIV Gag antigen (rBCG-SIV gag. We found that the incorporation of the synthetic NKT activating glycolipid α-galactosylceramide (α-GC into rBCG-SIV gag significantly enhanced CD8+ T cell responses against an immunodominant Gag epitope, compared to responses primed by unmodified rBCG-SIV gag. The abilities of structural analogues of α-GC to enhance CD8+ T cell responses to rBCG were compared in both wild type and partially humanized mice that express human CD1d molecules in place of mouse CD1d. These studies identified an α-GC analogue known as 7DW8-5, which has previously been used successfully as an adjuvant in non-human primates, as a promising compound for enhancing immunogenicity of antigens delivered by rBCG.vectors. Our findings support the incorporation of synthetic glycolipid activators of NKT cells as a novel approach to enhance the immunogenicity of rBCG-vectored antigens for induction of CD8+ T cell responses. The glycolipid adjuvant 7DW8-5 may be a promising candidate for advancing to non-human primate and human clinical studies for the development of HIV vaccines based on rBCG vectors.

  15. Persistence of Mycobacterium bovis Bacillus Calmette-Guerin (BCG) in White-tailed Deer (Odocoileus virginianus) After Oral or Parenteral Vaccination

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    Mycobacterium bovis is the cause of tuberculosis in cattle and a serious zoonotic pathogen, most commonly contracted through consumption of unpasteurized dairy products. To control this zoonosis, many countries have developed bovine tuberculosis eradication programs. Although relatively successful, ...

  16. Bacillus calmette-guerin cell wall cytoskeleton enhances colon cancer radiosensitivity through autophagy.

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    Yuk, Jae-Min; Shin, Dong-Min; Song, Kyoung-Sub; Lim, Kyu; Kim, Ki-Hye; Lee, Sang-Hee; Kim, Jin-Man; Lee, Ji-Sook; Paik, Tae-Hyun; Kim, Jun-Sang; Jo, Eun-Kyeong

    2010-01-01

    The cell wall skeleton of Mycobacterium bovis Bacillus Calmette-Guerin (BCG/CWS) is an effective antitumor immunotherapy agent. Here, we demonstrate that BCG/CWS has a radiosensitizing effect on colon cancer cells through the induction of autophagic cell death. Exposure of HCT116 colon cancer cells to BCG/CWS before ionizing radiation (IR) resulted in increased cell death in a caspase-independent manner. Treatment with BCG/CWS plus IR resulted in the induction of autophagy in colon cancer cells. Either the autophagy inhibitor 3-methyladenine or knockdown of beclin 1 or Atg7 significantly reduced tumor cell death induced by BCG/CWS plus IR, whereas the caspase inhibitor z-VAD-fmk failed to do so. BCG/CWS plus IR-mediated autophagy and cell death was mediated predominantly by the generation of reactive oxygen species (ROS). The c-Jun NH(2)-terminal kinase pathway functioned upstream of ROS generation in the induction of autophagy and cell death in HCT116 cells after co-treatment with BCG/CWS and IR. Furthermore, toll-like receptor (TLR) 2, and in part, TLR4, were responsible for BCG/CWS-induced radiosensitization. In vivo studies revealed that BCG/CWS-mediated radiosensitization of HCT116 xenograft growth is accompanied predominantly by autophagy. Our data suggest that BCG/CWS in combination with IR is a promising therapeutic strategy for enhancing radiation therapy in colon cancer cells through the induction of autophagy.

  17. Role of IL-10 in Urinary Bladder Carcinoma and Bacillus Calmette-Guerin Immunotherapy

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    Yi Luo

    2012-01-01

    Full Text Available Problem statement: Bladder cancer is a common urologic cancer and intravesical Mycobacterium bovis Bacillus Calmette-Guerin (BCG is the mainstay in the treatment of superficial bladder cancer. However, the current BCG therapy is not desirable with respect to its efficacy and side effects. Interleukin (IL-10, a T helper type (Th 2 cytokine, plays an important regulatory role in bladder cancer immunosurveillance and BCG immunotherapy. Therefore, blocking IL-10 activity could be beneficial for bladder cancer patients undergoing BCG therapy. Approach: Treatment with intravesical BCG in combination with systemic IL-10 monoclonal antibody (mAb specific for IL-10 neutralization or IL-10 receptor (IL-10R blockage has been evaluated in preclinical bladder cancer models. Results: Addition of anti-IL-10 neutralizing mAb or anti-IL-10R1 mAb enhances BCG induction of Th1 immune responses and anti-bladder cancer immunity. Conclusion/Recommendations: BCG immunotherapy of bladder cancer can be enhanced by addition of IL-10 blocking mAb. Future studies should aim to explore the mechanisms underlying the induction of enhanced antitumor immunity by BCG combination therapy and develop therapeutic regimens for clinical evaluation of the safety and efficacy of BCG combination therapy.

  18. FNAC of Bacillus- Calmette- Guerin lymphadenitis masquerading as Langerhans cell histiocytosis.

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    Gupta Nalini

    2004-12-01

    Full Text Available Abstract Bacillus Calmette Guerin (BCG lymphadenitis is a well known entity. Disseminated BCG infection usually presents as generalized lymphadenopathy, skin rash and hepatosplenomegaly and at times, can pose a diagnostic challenge to clinicians. There are only a few published studies on the cytological findings of BCG lymphadenitis. In this letter we report the fine needle aspiration cytology (FNAC of BCG lymphadenitis clinically masquerading as Langerhans cell histiocytosis (LCH. FNA smears showed sheets of foamy macrophages and many polymorphs in a dirty necrotic background with many macrophages as well as polymorphs showing negatively stained rod like structures within their cytoplasm. Zeihl Neelson stain revealed that these cells were heavily loaded with acid fast bacilli (AFB. In the index case, AFB were also seen within the cytoplasm of polymorphs, which has not been documented earlier in the literature.

  19. Cystectomy with orthotopic ileal neobladder reconstruction for treatment of bladder contracture after intravesical bacillus Calmette-Guerin therapy.

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    Vetorazzo Filho, José Eduardo; Bahia, Leandro Augusto Costa; Vedovato, Bruno César; Maron, Paulo Eduardo Goulart; Esteves, Paulo Ebert; Fernandes, Roni de Carvalho; Perez, Marjo Deninson Cardenuto

    2014-01-01

    Bladder cancer is an important health problem worldwide due to high prevalence rates and costs related to treatment. A reduction in recurrence rates has been observed since the introduction of adjuvant intravesical immunotherapy with bacillus Calmette-Guerin. There are mild complications that are easily solved by local measures and orientations. Bladder contracture, a rare and severe local complication, in some cases leading to disability, is observed primarily in patients in a maintenance program. In this article we reported the case of a male patient who underwent transurethral resection of the bladder because of a high-grade T1 urothelial carcinoma and developed this complication during treatment with bacillus Calmette-Guerin. For this reason he was submitted to cystoprostatectomy with orthotopic ileal neobladder reconstruction.

  20. Multiphase contrast-enhanced magnetic resonance imaging features of Bacillus Calmette-Guerin-induced granulomatous prostatitis in five patients

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    Kawada, Hiroshi; Kanematsu, Masayuki; Goshima, Satoshi; Kondo, Hiroshi; Watanabe, Haruo; Noda, Yoshifumi; Tanahashi, Yukichi; Kawai, Nobuyuki; Hoshi, Hiroaki [Gifu University Hospital, Gifu (Japan)

    2015-04-15

    To evaluate the multiphase contrast-enhanced magnetic resonance (MR) imaging features of Bacillus Calmette-Guerin (BCG)-induced granulomatous prostatitis (GP). Magnetic resonance images obtained from five patients with histopathologically proven BCG-induced GP were retrospectively analyzed for tumor location, size, signal intensity on T2-weighted images (T2WI) and diffusion-weighted images (DWI), apparent diffusion coefficient (ADC) value, and appearance on gadolinium-enhanced multiphase images. MR imaging findings were compared with histopathological findings. Bacillus Calmette-Guerin-induced GP (size range, 9-40 mm; mean, 21.2 mm) were identified in the peripheral zone in all patients. The T2WI showed lower signal intensity compared with the normal peripheral zone. The DWIs demonstrated high signal intensity and low ADC values (range, 0.44-0.68 x 10(-3) mm2/sec; mean, 0.56 x 10(-3) mm2/sec), which corresponded to GP. Gadolinium-enhanced multiphase MR imaging performed in five patients showed early and prolonged ring enhancement in all cases of GP. Granulomatous tissues with central caseation necrosis were identified histologically, which corresponded to ring enhancement and a central low intensity area on gadolinium-enhanced MR imaging. The findings on T2WI, DWI, and gadolinium-enhanced images became gradually obscured with time. Bacillus Calmette-Guerin-induced GP demonstrates early and prolonged ring enhancement on gadolinium-enhanced MR imaging which might be a key finding to differentiate it from prostate cancer.

  1. Primary tuberculosis of glans penis after intravesical Bacillus Calmette Guerin immunotherapy

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    V K Sharma

    2011-01-01

    Full Text Available A 55-year-old male with carcinoma in situ of urinary bladder was treated with weekly intravesical injections of Bacillus Calmette Guerin (BCG vaccine. Three days after the sixth injection, he developed low grade fever and multiple grouped punched out, 2-3 mm ulcers around meatus and corona glandis. In addition, multiple, firm, indurated, nontender papules and few deeper nodules were present on the proximal part of glans penis, along with bilateral enlarged, matted and nontender inguinal lymph nodes. There was no history suggestive of sexually transmitted diseases and high risk behavior. Chest X-ray was within normal limits, and Mantoux, Venereal Disease Research Laboratory (VDRL and HIV antibody tests were negative. The biopsy from the penile ulcer revealed epithelioid cell granuloma with Langhans giant cells. Fine needle aspiration cytology from the lymph node also revealed epithelioid cell granuloma and acid fast bacilli on Ziehl Neelsen′s stain. The tissue biopsy grew Mycobacterium tuberculosis. The BCG immunotherapy was stopped and patient was treated with four drug antitubercular therapy with isoniazid, rifampicin, ethambutol, and pyrazinamide in standard daily doses along with pyridoxine. The edema resolved and the ulcers started healing within 2 weeks, and at 6 weeks after starting antitubercular therapy almost complete healing occurred. To the best of our knowledge, we describe the first case of an Indian patient with BCG induced primary tuberculosis of penis after immunotherapy for carcinoma urinary bladder and review the previously described cases to increase awareness of this condition in dermatologists and venereologists.

  2. Discriminators of mouse bladder response to intravesical Bacillus Calmette-Guerin (BCG

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    Centola Michael

    2007-05-01

    Full Text Available Abstract Background Intravesical Bacillus Calmette-Guerin (BCG is an effective treatment for bladder superficial carcinoma and it is being tested in interstitial cystitis patients, but its precise mechanism of action remains poorly understood. It is not clear whether BCG induces the release of a unique set of cytokines apart from its pro-inflammatory effects. Therefore, we quantified bladder inflammatory responses and alterations in urinary cytokine protein induced by intravesical BCG and compared the results to non-specific pro-inflammatory stimuli (LPS and TNF-α. We went further to determine whether BCG treatment alters cytokine gene expression in the urinary bladder. Methods C57BL/6 female mice received four weekly instillations of BCG, LPS, or TNF-α. Morphometric analyses were conducted in bladders isolated from all groups and urine was collected for multiplex analysis of 18 cytokines. In addition, chromatin immune precipitation combined with real-time polymerase chain reaction assay (CHIP/Q-PCR was used to test whether intravesical BCG would alter bladder cytokine gene expression. Results Acute BCG instillation induced edema which was progressively replaced by an inflammatory infiltrate, composed primarily of neutrophils, in response to weekly administrations. Our morphological analysis suggests that these polymorphonuclear neutrophils are of prime importance for the bladder responses to BCG. Overall, the inflammation induced by BCG was higher than LPS or TNF-α treatment but the major difference observed was the unique granuloma formation in response to BCG. Among the cytokines measured, this study highlighted the importance of IL-1β, IL-2, IL-3, IL-4, IL-6, IL-10, IL-17, GM-CSF, KC, and Rantes as discriminators between generalized inflammation and BCG-specific inflammatory responses. CHIP/Q-PCR indicates that acute BCG instillation induced an up-regulation of IL-17A, IL-17B, and IL-17RA, whereas chronic BCG induced IL-17B, IL-17RA, and

  3. Routine Transurethral Biopsy of the Bladder is not Necessary to Evaluate the Response to Bacillus Calmette-guerin Therapy

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    Manabe,Daisuke

    2007-12-01

    Full Text Available We evaluated the need for transurethral biopsy at first follow-up after intravesical bacillus Calmette-Guerin (BCG therapy for superficial bladder cancer. The records of 84 patients with superficial bladder cancer who received a 6- or 8-week course of BCG were reviewed. Pathological results before BCG, cystoscopic findings, urinary cytology, and biopsy results for evaluation of BCG therapy were reviewed. All 19 patients with positive urinary cytology had evidence of positive bladder biopsy results. Fifty-three of 54 patients (98.1% with no visible recurrent tumor and negative urinary cytology demonstrated negative pathological results on bladder biopsy. When not found in conjunction with positive urinary cytology, erythematous mucosa on cystoscopy was not an indicator of tumor recurrence or residual cancer. In conclusion, routine transurethral biopsy of the bladder for evaluating the response to BCG intravesical therapy is not necessary in patients who have no visible tumor on cystoscopy and negative urinary cytology./

  4. Interferon-γ Added During Bacillus Calmette-Guerin Induced Dendritic Cell Maturation Stimulates Potent Th1 Immune Responses

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    Pestano Linda A

    2003-10-01

    Full Text Available Abstract Dendritic cells (DC are increasingly prepared in vitro for use in immunotherapy trials. Mature DC express high levels of surface molecules needed for T cell activation and are superior at antigen-presentation than immature DC. Bacillus Calmette-Guerin (BCG is one of several products known to induce DC maturation, and interferon (IFN-γ has been shown to enhance the activity of DC stimulated with certain maturation factors. In this study, we investigated the use of IFN-γ in combination with the powerful maturation agent, BCG. The treatment of immature DC with IFN-γ plus BCG led to the upregulation of CD54, CD80, and CD86 in comparison with BCG treatment alone. In MLR or recall immune responses, the addition of IFN-γ at the time of BCG-treatment did not increase the number of antigen-specific T cells but enhanced the development of IFN-γ-producing Th1 cells. In primary immune responses, on the other hand, BCG and IFN-γ co-treated DC stimulated higher proportions of specific T cells as well as IFN-γ secretion by these T cells. Thus the use of IFN-γ during BCG-induced DC maturation differentially affects the nature of recall versus naïve antigen-specific T-cell responses. IFN-γ co-treatment with BCG was found to induce IL-12 and, in some instances, inhibit IL-10 secretion by DC. These findings greatly enhance the potential of BCG-matured dendritic cells for use in cancer immunotherapy.

  5. Bacillus Calmette Guerin induces fibroblast activation both directly and through macrophages in a mouse bladder cancer model.

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    Catalina Lodillinsky

    Full Text Available BACKGROUND: Bacillus Calmette-Guerin (BCG is the most effective treatment for non-muscle invasive bladder cancer. However, a failure in the initial response or relapse within the first five years of treatment has been observed in 20% of patients. We have previously observed that in vivo administration of an inhibitor of nitric oxide improved the response to BCG of bladder tumor bearing mice. It was described that this effect was due to a replacement of tumor tissue by collagen depots. The aim of the present work was to clarify the mechanism involved in this process. METHODOLOGY/PRINCIPAL FINDINGS: We demonstrated that BCG induces NIH-3T3 fibroblast proliferation by activating the MAPK and PI3K signaling pathways and also differentiation determined by alpha-smooth muscle actin (alpha-SMA expression. In vivo, intratumoral inoculation of BCG also increased alpha-SMA and collagen expression. Oral administration of L-NAME enhanced the pro-fibrotic effect of BCG. Peritoneal macrophages obtained from MB49 tumor-bearing mice treated in vivo with combined treatment of BCG with L-NAME also enhanced fibroblast proliferation. We observed that FGF-2 is one of the factors released by BCG-activated macrophages that is able to induce fibroblast proliferation. The involvement of FGF-2 was evidenced using an anti-FGF2 antibody. At the same time, this macrophage population improved wound healing rate in normal mice and FGF-2 expression was also increased in these wounds. CONCLUSIONS/SIGNIFICANCE: Our findings suggest that fibroblasts are targeted by BCG both directly and through activated macrophages in an immunotherapy context of a bladder murine model. We also described, for the first time, that FGF-2 is involved in a dialog between fibroblasts and macrophages induced after BCG treatment. The fact that L-NAME administration improves the BCG effect on fibroblasts, NO inhibition, might represent a new approach to add to the conventional BCG therapy.

  6. Changes observed in multiparametric prostate magnetic resonance imaging characteristics correlate with histopathological development of chronic granulomatous prostatitis after intravesical Bacillus Calmette-Guerin therapy.

    Science.gov (United States)

    Logan, Jennifer K; Walton-Diaz, Annerleim; Rais-Bahrami, Soroush; Merino, Maria J; Turkbey, Baris; Choyke, Peter L; Pinto, Peter A

    2014-01-01

    Administration of Bacillus Calmette-Guerin (BCG) has been shown to cause granulomatous prostatitis, a rare inflammatory process that can be mistaken for prostate cancer. We present a case of a 78-year-old man on active surveillance for prostate cancer with a subsequent diagnosis of high-grade urothelial carcinoma. After intravesical BCG therapy, he developed chronic granulomatous prostatitis. We present serial magnetic resonance imaging and biopsy data demonstrating the time interval between BCG administration and the manifestation of chronic granulomatous prostatitis.

  7. Protective effect of melatonin against liver injury in mice induced by Bacillus Calmette-Guerin plus lipopolysaccharide

    Institute of Scientific and Technical Information of China (English)

    Hua Wang; Wei Wei; Yu-Xian Shen; Chen Dong; Ling-Ling Zhang; Ni-Ping Wang; Li Yue; Shu-Yun Xu

    2004-01-01

    AIM: To investigate the effects and mechanisms of melatonin on immunological liver injury in mice.METHODS: A model of liver injury was induced by tail veininjection of Bacillus Calmette Guerin (BCG) and lipopolysaccharide(LPS) in mice. Kupffer cells and hepatocytes were isolatedand cultured according to a modified two-step collagenaseperfusion technique. Levels of alanine aminotransferase(ALT), aspartate aminotransferase (AST) and nitric oxide(NO), content of malondiadehyde (MDA), activity of superoxidedismutase (SOD), were measured by biochemical methods.Tumor necrosis factor-α (TNF-α) activity was determinedby RIA. Interleukin (IL)-1 activity was measured by thymocyte proliferation bioassay. Hepatic tissue sections were stained with hematoxylin and eosin and examined under a lightmicroscope.RESULTS: Immunological liver injury induced by BCG+LPSwas successfully duplicated. Serum transaminase (ALT,AST) activities were significantly decreased by melatonin(0.25, 1.0, 4.0 mg/kg bm). Meanwhile, MDA content was decreased and SOD in liver homogenates was upregulated.Furthermore, pro-inflammatory mediators (TNF-α, IL-1, NO)in serum and liver homogenates were significantly reduced by melatonin. Histological examination demonstrated that melatonin could attenuate the area and extent of necrosis,reduce the immigration of inflammatory cells. In in vitro experiment, TNF-α was inhibited at the concentrations of10-8-10-6 mol/L of melatonin, while IL-1 production of Kupffer cells induced by LPS (5 μg/mL) was decreased only at theconcentration of 10-6 mol/L of melatonin, but no effect onNO production was observed. Immunological liver injury model in vitro was established by incubating hepatocyteswith BCG- and LPS-induced Kupffer cells. Activities of ALT,TNF-α, IL-1, and MDA in supernatant were significantlyincreased. Melatonin had little effect on the level of ALT,but reduced the content of TNF-α and MDA at concentrationsof 10-7-10-5 mol/L and decreased the content of IL-1

  8. Current clinical practice gaps in the treatment of intermediate- and high-risk non-muscle-invasive bladder cancer (NMIBC) with emphasis on the use of bacillus Calmette-Guerin (BCG): results of an international individual patient data survey (IPDS)

    NARCIS (Netherlands)

    Witjes, J.A.; Palou, J.; Soloway, M.; Lamm, D.; Kamat, A.M.; Brausi, M.; Persad, R.; Buckley, R.; Colombel, M.; Bohle, A.

    2013-01-01

    OBJECTIVES: To examine the management of intermediate- and high-risk non-muscle-invasive bladder cancer (NMIBC), particularly with regard to the use of bacillus Calmette-Guerin (BCG) therapy, in North America and Europe. To compare NMIBC management practices to European Association of Urology (EAU)

  9. The effect of age on the efficacy of maintenance bacillus calmette-guerin relative to maintenance epirubicin in patients with stage ta t1 urothelial bladder cancer: results from EORTC genito-urinary group study 30911

    NARCIS (Netherlands)

    Oddens, J.R.; Sylvester, R.J.; Brausi, M.A.; Kirkels, W.J.; Beek, C.; Andel, G. van; Reijke, T.M. de; Prescott, S.; Witjes, J.A.; Oosterlinck, W.

    2014-01-01

    BACKGROUND: Although maintenance bacillus Calmette-Guerin (BCG) is the recommended treatment in high-risk non-muscle-invasive bladder cancer (NMIBC), its efficacy in older patients is controversial. OBJECTIVE: To determine the effect of age on prognosis and treatment outcome in patients with stage T

  10. Bacillus calmette-guerin infection in NADPH oxidase deficiency: defective mycobacterial sequestration and granuloma formation.

    Directory of Open Access Journals (Sweden)

    Christine Deffert

    2014-09-01

    Full Text Available Patients with chronic granulomatous disease (CGD lack generation of reactive oxygen species (ROS through the phagocyte NADPH oxidase NOX2. CGD is an immune deficiency that leads to frequent infections with certain pathogens; this is well documented for S. aureus and A. fumigatus, but less clear for mycobacteria. We therefore performed an extensive literature search which yielded 297 cases of CGD patients with mycobacterial infections; M. bovis BCG was most commonly described (74%. The relationship between NOX2 deficiency and BCG infection however has never been studied in a mouse model. We therefore investigated BCG infection in three different mouse models of CGD: Ncf1 mutants in two different genetic backgrounds and Cybb knock-out mice. In addition, we investigated a macrophage-specific rescue (transgenic expression of Ncf1 under the control of the CD68 promoter. Wild-type mice did not develop severe disease upon BCG injection. In contrast, all three types of CGD mice were highly susceptible to BCG, as witnessed by a severe weight loss, development of hemorrhagic pneumonia, and a high mortality (∼ 50%. Rescue of NOX2 activity in macrophages restored BCG resistance, similar as seen in wild-type mice. Granulomas from mycobacteria-infected wild-type mice generated ROS, while granulomas from CGD mice did not. Bacterial load in CGD mice was only moderately increased, suggesting that it was not crucial for the observed phenotype. CGD mice responded with massively enhanced cytokine release (TNF-α, IFN-γ, IL-17 and IL-12 early after BCG infection, which might account for severity of the disease. Finally, in wild-type mice, macrophages formed clusters and restricted mycobacteria to granulomas, while macrophages and mycobacteria were diffusely distributed in lung tissue from CGD mice. Our results demonstrate that lack of the NADPH oxidase leads to a markedly increased severity of BCG infection through mechanisms including increased cytokine

  11. SAPHO syndrome with bacillus Calmette-Guerin (BCG) immunotherapy for bladder cancer.

    Science.gov (United States)

    Matsumaru, Katsuhiko; Nagai, Kazuki; Murakami, Takayuki; Andoh, Kazuo

    2010-08-31

    The authors describe a case of SAPHO syndrome with bacillus Calmette-Guérin (BCG) immunotherapy for bladder cancer. The patient had undergone transurethral resection (TUR) and was treated with BCG immunotherapy following TUR. Two years after treatment for bladder cancer, the patient had palmoplantar pustulosis, and in the past 1 month suffered from pain localised to the anterior chest wall. The bone scintigraphy showed a strong focal enrichment in the right chest wall, suggesting spondyloarthropathy rather than malignant disease. On the basis of clinical and scintigraphy findings, SAPHO syndrome was diagnosed. The patient was treated with topical therapy and non-steroidal anti-inflammatory drugs and symptoms improved. The authors suggest that SAPHO syndrome might be caused by an association with BCG immunotherapy.

  12. Molecular networks discriminating mouse bladder responses to intravesical bacillus Calmette-Guerin (BCG, LPS, and TNF-α

    Directory of Open Access Journals (Sweden)

    Dozmorov Igor

    2008-02-01

    Full Text Available Abstract Background Despite being a mainstay for treating superficial bladder carcinoma and a promising agent for interstitial cystitis, the precise mechanism of Bacillus Calmette-Guerin (BCG remains poorly understood. It is particularly unclear whether BCG is capable of altering gene expression in the bladder target organ beyond its well-recognized pro-inflammatory effects and how this relates to its therapeutic efficacy. The objective of this study was to determine differentially expressed genes in the mouse bladder following chronic intravesical BCG therapy and to compare the results to non-specific pro inflammatory stimuli (LPS and TNF-α. For this purpose, C57BL/6 female mice received four weekly instillations of BCG, LPS, or TNF-α. Seven days after the last instillation, the urothelium along with the submucosa was removed from detrusor muscle and the RNA was extracted from both layers for cDNA array experiments. Microarray results were normalized by a robust regression analysis and only genes with an expression above a conditional threshold of 0.001 (3SD above background were selected for analysis. Next, genes presenting a 3-fold ratio in regard to the control group were entered in Ingenuity Pathway Analysis (IPA for a comparative analysis in order to determine genes specifically regulated by BCG, TNF-α, and LPS. In addition, the transcriptome was precipitated with an antibody against RNA polymerase II and real-time polymerase chain reaction assay (Q-PCR was used to confirm some of the BCG-specific transcripts. Results Molecular networks of treatment-specific genes generated several hypotheses regarding the mode of action of BCG. BCG-specific genes involved small GTPases and BCG-specific networks overlapped with the following canonical signaling pathways: axonal guidance, B cell receptor, aryl hydrocarbon receptor, IL-6, PPAR, Wnt/β-catenin, and cAMP. In addition, a specific detrusor network expressed a high degree of overlap with the

  13. Therapy of Venezuelan patients with severe mucocutaneous or early lesions of diffuse cutaneous leishmaniasis with a vaccine containing pasteurized Leishmania promastigotes and bacillus Calmette-Guerin: preliminary report

    Directory of Open Access Journals (Sweden)

    Jacinto Convit

    2004-02-01

    Full Text Available Severe mucocutaneous (MCL and diffuse (DCL forms of American cutaneous leishmaniasis (ACL are infrequent in Venezuela. Chemotherapy produces only transitory remission in DCL, and occasional treatment failures are observed in MCL. We have evaluated therapy with an experimental vaccine in patients with severe leishmaniasis. Four patients with MCL and 3 with early DCL were treated with monthly intradermal injections of a vaccine containing promastigotes of Leishmania (Viannia braziliensis killed by pasteurization and viable Bacillus Calmette- Guerin. Clinical and immunological responses were evaluated. Integrity of protein constituents in extracts of pasteurized promastigotes was evaluated by gel electrophoresis. Complete remission of lesions occurred after 5-9 injections in patients with MCL or 7-10 injections in patients with early DCL. DCL patients developed positive skin reactions, average size 18.7 mm. All have been free of active lesions for at least 10 months. Adverse effects of the vaccine were limited to local reactivity to BCG at the injection sites and fever in 2 patients. Extracts of pasteurized and fresh promastigotes did not reveal differences in the integrity of protein components detectable by gel electrophoresis. Immunotherapy with this modified vaccine offers an effective, safe option for the treatment of patients who do not respond to immunotherapy with vaccine containing autoclaved parasites or to chemotherapy .

  14. Adaptive immunity in the colostrum-deprived calf: response to early vaccination with Mycobacterium bovis strain bacille Calmette Guerin and ovalbumin.

    Science.gov (United States)

    Nonnecke, B J; Waters, W R; Goff, J P; Foote, M R

    2012-01-01

    Responses of the newborn calf to vaccination are frequently characterized by marginal antibody (Ab) responses. The present study evaluated effects of colostrum ingestion on the adaptive immune response of the preruminant calf to early vaccination. Colostrum-fed (CF) and colostrum-deprived (CD) calves were vaccinated at 2 d of age with Mycobacterium bovis, Pasteur strain of bacille Calmette Guerin (BCG), and ovalbumin (OVA) to track development of the adaptive immune response during the first 8 wk of life. Dams were also vaccinated with BCG prepartum. At wk 0, serum IgG(1), IgG(2), IgA, and IgM were elevated in CF calves, with IgG(1) predominating. In these calves, IgG(2), IgA, and IgM concentrations decreased with age. The CD calves, in contrast, had very low or undetectable serum immunoglobulin concentrations at wk 0 followed by an age-related increase in IgG(1), IgG(2), and IgM concentrations, suggesting endogenous production of these immunoglobulin classes. Immunoblot and ELISA analyses of Ab response to BCG vaccination indicated that colostrum ingestion was associated with measurable serum anti-mycobacterial Ab in CF calves during the first month postpartum, with substantially lower levels at 7 wk of age. Although mycobacteria-specific Ab was undetectable in CD calves at wk 0, it was present at 4 and 7 wk of age, suggesting that these calves, unlike CF calves, were capable of generating an Ab response to BCG vaccination. Antibody responses of CF and CD calves to vaccination with OVA, an antigen not present in the natural environment of dairy cattle, were of comparable magnitude and characterized by a progressive increase in Ab levels from birth (wk 0) to 7 wk of age. The disparate Ab responses of CF calves to BCG and OVA suggest that maternal antigenic experience or exposure influences Ab responses of the colostrum-fed preruminant calf to early vaccination. Ex vivo, antigen [OVA and M. bovis-derived purified protein derivative (PPDb)]-induced IFN-γ and nitric

  15. Neonatal bacillus Calmette-Guerin vaccination inhibits de novo allergic inflammatory response in mice via alteration of CD4+CD25+T-regulatory cells

    Institute of Scientific and Technical Information of China (English)

    Qian LI; Hua-hao SHEN

    2009-01-01

    Aim: The hygiene hypothesis suggests a lack of bacterial infections would favor the development of allergic diseases. My-cobacterium bovis bacille Calmette-Guerin (BCG) infection can inhibit allergen-induced asthma reactions, but the underly-hag mechanism of this infection on the immunological responses is unclear. T-regulatory (Treg) cells are thought to play a role as a crucial immunoregulatory cells that are capable of regulating adaptive immune responses. We conducted this study to investigate whether the protective effect of the BCG vaccination on allergic pulmonary inflammation is associated with the alteration of CD4+CD25+ Treg cells in a murine asthma model and the mechanisms of Treg cells. Methods: Newborn C57BL/6 mice were vaccinated 3 times with BCG on d 0, 7, and 14 and subsequently sensitized and challenged with ovalbumin. Eosinophil infiltration was investigated. The frequencies of spleen CD4+CD25+ Treg cells and the expression of specific transcriptional factor Foxp3 were assayed. The cytotoxic lymphocyte associated antigen (CTLA)-4 expression and cytokine interleukin-10 (IL-10) and transforming growth factor-β (TGF-β) levels were measured. Results: We showed that treatment of mice with BCG inhibited de novo allergic inflammatory response in a mouse model of asthma. BCG treatments are associated with the increase of CD4+CD25+ Treg cells and Foxp3 expression, accompanied by an increased CTLA-4 expression and cytokine IL-10 and TGF-β levels (P<0.05). Conclusion: Neonatal BCG vaccinations ameliorate de novo local eosinophilic inflammation induced by allergen and in-crease the numbers of CD4+CD25+ Treg cells and Foxp3 expression. The cell-cell contact inhibition and regulatory cytokine production may be involved in the regulatory mechanism.

  16. Prime-boost vaccination with Bacillus Calmette Guerin and a recombinant adenovirus co-expressing CFP10, ESAT6, Ag85A and Ag85B of Mycobacterium tuberculosis induces robust antigen-specific immune responses in mice.

    Science.gov (United States)

    Li, Wu; Li, Min; Deng, Guangcun; Zhao, Liping; Liu, Xiaoming; Wang, Yujiong

    2015-08-01

    Tuberculosis (TB) remains to be a prevalent health issue worldwide. At present, Mycobacterium bovis Bacillus Calmette Guerin (BCG) is the singular anti-TB vaccine available for the prevention of disease in humans; however, this vaccine only provides limited protection against Mycobacterium tuberculosis (Mtb) infection. Therefore, the development of alternative vaccines and strategies for increasing the efficacy of vaccination against TB are urgently required. The present study aimed to evaluate the ability of a recombinant adenoviral vector (Ad5-CEAB) co-expressing 10-kDa culture filtrate protein, 6-kDa early-secreted antigenic target, antigen 85 (Ag85)A and Ag85B of Mtb to boost immune responses following primary vaccination with BCG in mice. The mice were first subcutaneously primed with BCG and boosted with two doses of Ad5-CEAB via an intranasal route. The immunological effects of Ad5-CEAB boosted mice primed with BCG were then evaluated using a series of immunological indexes. The results demonstrated that the prime-boost strategy induced a potent antigen-specific immune response, which was primarily characterized by an enhanced T cell response and increased production of cytokines, including interferon-γ, tumor necrosis factor-α and interleukin-2, in mice. In addition, this vaccination strategy was demonstrated to have an elevated humoral response with increased concentrations of antigen-specific bronchoalveolar lavage secretory immunoglobulin (Ig)A and serum IgG in mice compared with those primed with BCG alone. These data suggested that the regimen of subcutaneous BCG prime and mucosal Ad5-CEAB boost was a novel strategy for inducing a broad range of antigen-specific immune responses to Mtb antigens in vivo, which may provide a promising strategy for further development of adenoviral-based vaccine against Mtb infection.

  17. Clinical analysis of 23 cases of disseminated bacillus Calmette-Guerin disease%播散性卡介菌病23例分析

    Institute of Scientific and Technical Information of China (English)

    卢水华; 李涛; 席秀红; 郭建; 黄富礼; 张文宏; 刘旭晖; 范小勇

    2013-01-01

    Objective To explore the clinical characteristics,diagnosis,treatment and prognosis of disseminated bacillus Calmette-Guerin (BCG) disease.Methods A retrospective study was performed on 23 children diagnosed with disseminated BCG disease at Shanghai Public Health Clinical Center from March 2006 to June 2011.The diagnosis,clinical features,underlying immunodeficiency,treatment and prognosis were evaluated.Results Among the 23 children,13 were male and 10 were female.The age of patients was from 2 months to 5 years old when diagnosed (median age,14.5 months).Clinical manifestations included fever in 23 cases,night sweats in 15.Thoracic and abdominal computed tomography of the 23 cases showed infiltrations or miliary lesion of the lungs with hilar,mediastinal and abdominal lymphadenopathy.Tuberculin skin tests (TST) were positive in 60.9% (14/23),and interferon gamma release assay (IGRA) was positive in 23.1% (3/13).Among the 23 cases,14 cases were diagnosed with primary combined immunodeficiency disease.Co-infections were observed in 14 out of 23 patients.All patients were given 2-4 antituberculous agents,including 12 severe cases with additional linezolid,and 16 cases with thymosin.During follow-up,there were 7 deaths (30.4%).Conclusions Clinical diagnostic criteria of disseminated BCG disease needs to be advised.Disseminated BCG disease tends to occur in children with immunodeficiency.The prognosis is poor and treatment options is limited at present.%目的 探讨播散性卡介菌病的临床特点、诊断、治疗和预后.方法 收集2006年3月至2011年6月在上海市公共卫生临床中心诊断的播散性卡介菌病患儿23例,评价诊断标准,分析临床特点、免疫缺陷、治疗和预后.结果 23例患儿中,男13例,女10例.首诊年龄为2个月~5岁,中位年龄14.5个月.临床表现发热23例、盗汗15例.23例患儿均进行胸腹部CT检查,均发现肺内实质浸润或粟粒样病灶伴肺门、纵隔

  18. Transdifferentiation of Small Cell Carcinoma of the Urinary Bladder from Urothelial Carcinoma after Transurethral Resection of a Bladder Tumor, Intravesical Bacillus Calmette-Guerin Instillation, and Chemotherapy: A Case Report

    Directory of Open Access Journals (Sweden)

    Kento Morozumi

    2016-11-01

    Full Text Available A 73-year-old male underwent transurethral resection of a bladder tumor in August 2010 and April 2011. Pathological examination revealed urothelial carcinoma. After the surgery, chemotherapy and intravesical Bacillus Calmette-Guerin instillation were performed. In September 2014, he once again underwent transurethral resection of the bladder tumor for recurrence, and was again diagnosed with urothelial carcinoma, pT2, by pathological examination. After neoadjuvant chemotherapy, radical cystectomy for tumor recurrence was performed. Pathological examination at this time revealed small cell carcinoma, pT3N0. It is rare for urothelial carcinoma to change to small cell carcinoma, and the mechanism and cause of this change are still unknown. In this case report, we discuss what causes small cell carcinoma of the urinary bladder and review the literature regarding its origin.

  19. Suppurative supraclavicular bacille calmette-guerine lymphadenitis - A case report, awareness and management options

    OpenAIRE

    U S Udgaonkar; Patil, S S; Rekha, V.B.; Shah, S.

    2015-01-01

    Diagnosis of Bacille calmette-guerine (BCG) adenitis is clinical. Conventional laboratory tests do not differentiate BCG adenitis from tuberculous adenitis. We report a case of a 3-month-old healthy baby presenting with suppurative BCG adenitis. FNAC revealed AFB on ZN-Staining, later confirmed to be Mycobacterium bovis by multiplex PCR. The treatment of suppurative BCG adenitis is needle aspiration. Anti-tubercular treatment is unwarranted.

  20. Suppurative supraclavicular bacille calmette-guerine lymphadenitis - A case report, awareness and management options

    Directory of Open Access Journals (Sweden)

    U S Udgaonkar

    2015-01-01

    Full Text Available Diagnosis of Bacille calmette-guerine (BCG adenitis is clinical. Conventional laboratory tests do not differentiate BCG adenitis from tuberculous adenitis. We report a case of a 3-month-old healthy baby presenting with suppurative BCG adenitis. FNAC revealed AFB on ZN-Staining, later confirmed to be Mycobacterium bovis by multiplex PCR. The treatment of suppurative BCG adenitis is needle aspiration. Anti-tubercular treatment is unwarranted.

  1. Investigation of the quality of Bacillus Calmette-Guerin vaccination and its related factors in 1533 infants%1533名婴幼儿卡介苗接种质量及其相关影响因素研究

    Institute of Scientific and Technical Information of China (English)

    时文明; 史太平; 朋文佳; 顾昕

    2016-01-01

    Objective:To investigate the vaccination quality of Bacillus Calmette-Guerin ( BCG ) in infants, analyze its related impacting factors and improve the effects of vaccination. Methods:The results of tuberculin( PPD) test in 1533 infants vaccinated with BCG and its related factors from Institute of Preventive Vaccination of Changzhou were analyzed,which was used to evaluate the quality of BCG vaccination. Results:The BCG scar rate and PPD positive rate were 94. 59% and 96. 09%,respectively. The differences of PPD positive rate in different gender,household register,birthweight and age were not statistically significant(P>0. 05). The PPD positive rates in infants with 3 to 5 mm or more than 5 mm of BCG scar diameter were significantly higher than that in infants with less than 3 mm of BCG scar diameter(P 0.05).卡痕均径3~5 mm和>5 mm的婴幼儿PPD试验阳转率均明显高于卡痕<3 mm者(P<0.01).结论:常州市结核病防治研究所BCG接种质量较好,达到国家基础免疫标准.性别、户籍地、出生体质量、PPD试验年龄对BCG阳转率均无明显影响,重视BCG质量和BCG接种技术对提高接种效果作用显著.

  2. Study on pre-immunization with Bacillus Calmette-Guerin (BCG) on Candida albicans infection in mice%卡介苗免疫预防小鼠白色念珠菌感染的实验研究

    Institute of Scientific and Technical Information of China (English)

    李丽波; 王玉祥

    2009-01-01

    目的 探讨卡介苗(BCG)预先免疫对小鼠白色念珠菌感染的影响.方法 采用BCG或灭菌生理盐水皮内注射预先免疫2周,然后由尾静脉注射白色念珠菌进行攻击,观察小鼠死亡率及计算存活小鼠肾组织带菌量.结果 BCG免疫组存活时间长于生理盐水免疫组(P<0.01);存活小鼠肾组织菌落计数:BCG免疫组远少于省里盐水免疫组(P<0.01).结论 BCG对小鼠白色念珠菌感染具有很好的保护作用.%Objective To investigate the effect of preimmunization with Bacillus Calmette-Guerin (BCG) on Candida albicans infection in mice. Methods BCG or 0.9% sterile saline was injected intracutaneously to ICR mice for preimmunization for two weeks before inoculation of Candida albicans by vena caudalis injection, then mortality rate of mice was observed and Candida albicans in mice kidney was examined. Results Survival time in BCG preimmunization group was longer than that in 0.9% sterile saline group (P<0.01); colony count of Candida albicans in mice kidney in BCG preimmunization group was less than that in saline group (P < 0.01). Conclusions BCG pre-immunization has protective effect from Candida albicans infection in mice.

  3. Prophylaxis Effect of Polysaccharide Nucleic Acid Fraction of Bacillus Calmette Guerin on Condyloma Acuminatum Recurrence: a Meta-analysis%卡介菌多糖核酸预防尖锐湿疣复发的Meta分析

    Institute of Scientific and Technical Information of China (English)

    吴丽峰; 吴大兴; 普雄明

    2012-01-01

    Objective To demonstrate the prophylaxis effect of polysaecharide nucleic acid fraction of bacillus calmette guerin (BCG-PSN) on condyloma acuminatum recurrence. Methods Clinical randomized controlled trials about BCG-PSN in treating condyloma acuminatum which met the criteria were searched. A Meta-analysis were performed using Statal 1.0 software to compare the prophylaxis effect between BCG-PSN combined CO2 laser and single CO2 laser therapy on condyloma acuminatum recurrence. Results A total of 11 studies were included, the results of Meta-analysis showed that condyloma acuminatum recurrence rate was significantly lower on BCG-PSN combined CO2 laser group than on single CO2 laser therapy group at 3 & 6 mon-thes, the relative risk (RR) were 0.41 (95%CI,0.33-0.51) and 0.23 (95%CI,0.12-0.44), respectively. Conclusion Compared with single CO2 laser therapy, BCG-PSN combined CO2 laser is associated with a low recurrence rate, and is safe and well-tolerated.%目的 探讨卡介菌多糖核酸预防尖锐湿疣复发的效果.方法 检索符合标准的关于卡介菌多糖核酸治疗尖锐湿疣的随机对照试验,应用Stata11.0软件进行Meta分析,比较卡介菌多糖核酸联合CO2激光与单纯CO2激光治疗尖锐湿疣的复发率.结果 共纳入相关文献11篇,Meta分析结果显示在治疗后3个月及6个月,卡介菌多糖核酸联合CO2激光治疗尖锐湿疣的复发率均显著低于单纯CO2激光治疗组,相对危险度RR分别为0.41 (95%CI,0.33~0.51)和0.23 (95%CI,0.12~0.44).结论 卡介菌多糖核酸联合CO2激光治疗尖锐湿疣复发率低,安全好,且患者耐受性好.

  4. Altered Memory T-Cell Responses to Bacillus Calmette-Guerin and Tetanus Toxoid Vaccination and Altered Cytokine Responses to Polyclonal Stimulation in HIV-Exposed Uninfected Kenyan Infants.

    Science.gov (United States)

    Garcia-Knight, Miguel A; Nduati, Eunice; Hassan, Amin S; Gambo, Faith; Odera, Dennis; Etyang, Timothy J; Hajj, Nassim J; Berkley, James Alexander; Urban, Britta C; Rowland-Jones, Sarah L

    2015-01-01

    Implementation of successful prevention of mother-to-child transmission of HIV strategies has resulted in an increased population of HIV-exposed uninfected (HEU) infants. HEU infants have higher rates of morbidity and mortality than HIV-unexposed (HU) infants. Numerous factors may contribute to poor health in HEU infants including immunological alterations. The present study assessed T-cell phenotype and function in HEU infants with a focus on memory Th1 responses to vaccination. We compared cross-sectionally selected parameters at 3 and 12 months of age in HIV-exposed (n = 42) and HU (n = 28) Kenyan infants. We measured ex vivo activated and bulk memory CD4 and CD8 T-cells and regulatory T-cells by flow cytometry. In addition, we measured the magnitude, quality and memory phenotype of antigen-specific T-cell responses to Bacillus Calmette-Guerin and Tetanus Toxoid vaccine antigens, and the magnitude and quality of the T cell response following polyclonal stimulation with staphylococcal enterotoxin B. Finally, the influence of maternal disease markers on the immunological parameters measured was assessed in HEU infants. Few perturbations were detected in ex vivo T-cell subsets, though amongst HEU infants maternal HIV viral load positively correlated with CD8 T cell immune activation at 12 months. Conversely, we observed age-dependent differences in the magnitude and polyfunctionality of IL-2 and TNF-α responses to vaccine antigens particularly in Th1 cells. These changes mirrored those seen following polyclonal stimulation, where at 3 months, cytokine responses were higher in HEU infants compared to HU infants, and at 12 months, HEU infant cytokine responses were consistently lower than those seen in HU infants. Finally, reduced effector memory Th1 responses to vaccine antigens were observed in HEU infants at 3 and 12 months and higher central memory Th1 responses to M. tuberculosis antigens were observed at 3 months only. Long-term monitoring of vaccine efficacy

  5. p53 Status correlates with the risk of recurrence in non-muscle invasive bladder cancers treated with Bacillus Calmette-Guerin: a meta-analysis.

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    Xiaofeng Zhou

    Full Text Available Published studies have yielded inconsistent results on the relationship between p53 status and the prognosis of non-muscle invasive bladder cancer (NMIBC treated with Bacillus Calmette-Guérin (BCG intravesical therapy. Therefore, we performed a meta-analysis to evaluate the prognostic value of p53 in NMIBC treated with BCG.We systematically searched for relevant literature in PubMed, EMBASE, CNKI, and Chinese Wanfang databases. Hazard ratios (HRs with 95% confidence intervals (CIs were combined as the effect size (ES across studies for recurrence-free survival (RFS and progression-free survival (PFS.A total of 11 studies, consisting of 1,049 participants, met the criteria. Overall, there was no clear relationship between p53 status and RFS or PFS for NMIBC patients treated with BCG (HR: 1.40, 95% CI: 0.91-2.16; HR: 1.37, 95% CI: 0.90-2.09, respectively. Obvious heterogeneity was observed across the studies (I2 = 69.5%, P = 0.001; I2 = 44.7%, P = 0.081, respectively. In stratified analysis by region, p53 overexpression was a predictor of poor RFS in Asian populations (HR: 1.57, 95% CI: 1.08-2.27. In addition, after excluding the studies that possibly contributed to the heterogeneity by the Galbraith plot, the overall association for RFS became statistically significant (HR: 1.38 95% CI: 1.08-1.77 without evidence of heterogeneity (I2 = 0.0%, P = 0.499.This meta-analysis suggests that p53 overexpression in NMIBC patients treated with BCG may be associated with RFS, especially in Asian populations. Because of the heterogeneity and other limitations, further studies with rigid criteria and large populations are still warranted to confirm our findings.

  6. Polysaccharide Nucleic Acid of Bacillus Calmette Guerin Modulates Th1/Th2 Cytokine Gene Expression in Lipopolysaccharide-Induced Mastitis in Rats

    Institute of Scientific and Technical Information of China (English)

    MIAO Jin-feng; ZHANG Yuan-shu; HUANG Guo-qing; MA Hai-tian; ZOU Si-xiang; ZHU Yu-min

    2009-01-01

    The aim of this study was to evaluate, in rats, the changes in the T helper type 1 (Th 1)/Th2 radio in mammary glands after an intramammary infusion of lipopolysaccharide (LPS) and to characterize the moderating effects of the polysaecharide nucleic acid of Bacillus Calraette Guerin (BCG-PSN) on the mammary gland. In the control group, the levels of IL-2 and INF-γ, mRNA expression increased, whereas IL-4 mRNA expression decreased after LPS challenge. As a consequence, the INF-γ/IL-4 mRNA ratio was significantly higher at 3, 6, and 9 h post-infusion (PI) compared to the control value (O h; P<0.01).BCG-PSN increased mRNA expression of both INF-γ' and IL-4 before infusion of LPS. LPS challenge significantly the reduced Th1/Th2 eytokine ratio due to Th1 cytokine IFN-γ suppression and Th2 cytokine IL-4 upregulation compared with the control group. A significant reduction ofN-acety1-β-D-glucosaminidase (NAGase) was observed at 24 h PI in the BCG-PSN treatment group compared to the control group (P<0.05). Thus, it was demonstrated that level of BCG-PSN might change the Th1/Th2 ratio mainly by enhancing the Th2 immune response. This is the first report of a Th1/Th2 change induced by coliform mastitis and characterization of the effect of BCG-PSN on mammary gland inflammation. This study makes a better understanding of the mechanisms of coliform mastitis and provides a putative novel strategy for the prevention and/or treatment of mastitis.

  7. Re: Sequential Combination of Mitomycin C Plus Bacillus Calmette-Guerin (BCG Is More Effective but More Toxic Than BCG Alone in Patients with Non–Muscle-Invasive Bladder Cancer in Intermediate-and High-Risk Patients: Final Outcome of CUETO 93009, A Randomized Prospective Trial

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    Eduardo Solsona

    2015-03-01

    Full Text Available EAU Guideline recommendation in non-muscle invasive bladder cancer (NMIBC is that patients who have intermediate or high risk for recurrence and intermediate risk for progression should receive early single dose intravesical chemotherapy followed by maintenance or a minimum of 1 year of BCG. Intravesical Mitomycin C (MMC plus Bacillus Calmette-Guerin (BCG treatment schemes were studied. However, MMC+BCG were not found to be superior to BCG alone (1,2. In the present study, authors conducted a randomized prospective trial on combination of MMC+BCG (n=192 or BCG alone (n=190. EORTC definition of NMIBC intermediate and high-risk patientswere included in the study. Unlike previous reported studies, disease-free interval at 5 years for MMC+BCG was found to be significantly better (HR: 0.57; 95% CI, 0.39 -0,83; p=0.003 than BCG alone. In an interim analysis, excessive toxicity was observed in MMC+BCG than BCG alone group. Consequently MMC dose was reduced from 30 mg to 10 mg. However, toxicity remained higher in the MMC+BCG group. Especially in EORTC highrisk NMIBCs, MMC+BCG is better than BCG alone, but with worse toxicity. In conclusion, despite some limitations, the results of Solsona et al. provided a new potential bladder-sparing management alternative, but it has higher toxicity. Additional studies are required to confirm these findings and availability of a less toxic intravesical chemotherapeutic agent.

  8. Revisiting the structure of the anti-neoplastic glucans of Mycobacterium bovis Bacille Calmette-Guerin. Structural analysis of the extracellular and boiling water extract-derived glucans of the vaccine substrains.

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    Dinadayala, Premkumar; Lemassu, Anne; Granovski, Pierre; Cérantola, Stéphane; Winter, Nathalie; Daffé, Mamadou

    2004-03-26

    The attenuated strain of Mycobacterium bovis Bacille Calmette-Guérin (BCG), used worldwide to prevent tuberculosis and leprosy, is also clinically used as an immunotherapeutic agent against superficial bladder cancer. An anti-tumor polysaccharide has been isolated from the boiling water extract of the Tice substrain of BCG and tentatively characterized as consisting primarily of repeating units of 6-linked-glucosyl residues. Mycobacterium tuberculosis and other mycobacterial species produce a glycogen-like alpha-glucan composed of repeating units of 4-linked glucosyl residues substituted at some 6 positions by short oligoglucosyl units that also exhibits an anti-tumor activity. Therefore, the impression prevails that mycobacteria synthesize different types of anti-neoplastic glucans or, alternatively, the BCG substrains are singular in producing a unique type of glucan that may confer to them their immunotherapeutic property. The present study addresses this question through the comparative analysis of alpha-glucans purified from the extracellular materials and boiling water extracts of three vaccine substrains. The polysaccharides were purified, and their structural features were established by mono- and two-dimensional NMR spectroscopy and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of the enzymatic and chemical degradation products of the purified compounds. The glucans isolated by the two methods from the three substrains of BCG were shown to exhibit identical structural features shared with the glycogen-like alpha-glucan of M. tuberculosis and other mycobacteria. Incidentally, we observed an occasional release of dextrans from Sephadex columns that may explain the reported occurrence of 6-substituted alpha-glucans in mycobacteria.

  9. The effects of polysaccharide nucleic acid fractiion of bacillus calmette guerin and specific immunotherapy on pulmonary function and serum IgE in asthmatic children%卡介菌多糖核酸和特异性免疫治疗对哮喘儿童肺功能和血清IgE的影响

    Institute of Scientific and Technical Information of China (English)

    黄李平; 刘战军; 汪清

    2011-01-01

    目的 研究卡介菌多糖核酸(BCG-PNA)和特异性免疫治疗(SIT)对哮喘儿童肺功能和血清IgE的影响.方法 用BCG-PNA和SIT治疗42例哮喘儿童,治疗前后检测肺功能,用酶联免疫吸附试验检测血清IgE水平.结果 BCG-PNA、SIT能明显改善哮喘儿童肺功能FVC、FEV1、V25等指标,BCG-PNA还能降低血清IgE水平.两者联合治疗比SIT治疗降低血清IgE更明显.结论 BCG-PNA和SIT能改善哮喘儿童肺功能,两者联合治疗对降低血清IgE有协同作用.%Objective This research aimed to explore the effects of polysaccharide nucleic acid fraction of Bacillus calmette Guerin (BCG-PNA) and specific immunotherapy (SIT) on pulmonary function and serum IgE in asthmatic children. Methods Before and after treatments with BCG-PNA and SIT, pulmonary function of 40 asthmatic children was analyzed. The serum IgE was detected by ELISA. Results BCG-PNA and SIT could improve FVC, FEV1 and V25 of pulmonary function indexes. BCG-PNA decreased serum IgE. Combination of these two treatments reduced serum IgE compared with SIT. Conclusion Both BCG-PNA and SIT can improve pulmonary function of asthmatic children. Combination of them has stronger effect in reducing serum IgE.

  10. Immunoscintigraphy in the detection of tuberculosis with radiolabelled antibody fragment against Mycobacterium bovis bacillus Calmette-Guerin. A preliminary study in a rabbit model

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    Lee, J.D.; Park, C.Y.; Yoo, H.S.; Lee, J.T. (Yonsei Univ., Seoul (Korea, Republic of). Dept. of Diagnostic Radiology); Shin, K.H. (Yonsei Univ., Seoul (Korea, Republic of). Dept. of Orthopedic Surgery); Cho, S.N.; Shin, J.S. (Yonsei Univ., Seoul (Korea, Republic of). Dept. of Microbiology); Lee, M.G. (Yonsei Univ., Seoul (Korea, Republic of). Dept. of Dermatology); Yang, W.I. (Yonsei Univ., Seoul (Korea, Republic of). Dept. of Pathology); Awh, O.D. (Korea Atomic Energy Research Inst., Seoul (Korea, Republic of). Dept. of Reactor Isotopes)

    1992-12-01

    Immunoscintigraphy with radiolabelled monoclonal antibodies is widely used to detect solid tumours, but only a few trials have been carried out concerning the specific in vivo localization of an inflammatory process. The purpose of this study was to investigate the detectability of tuberculous foci utilizing this method with radiolabelled bacille Colmette-Guerin (BCG)-specific F(ab')[sub 2] in rabbits. All of the tuberculous lesions (n=8) were clearly visualized on serial scintigraphy for up to 48 h after injection of the antibody. Immunohistochemical and Ziel-Neelson staining of the tuberculous lesions confirmed the presence of the tuberculous antigens and bacilli. It failed to demonstrate any sustained retention of the BCG-specific antibody fragment in the control group with syphilitic orchitis (n=2). Therefore, the specific in vivo localization of tuberculosis is feasible by immunoscintigraphy. (orig.).

  11. Late-onset granulomatous prostatitis following intravesical bacille Calmette-Guerin therapy: case report.

    Science.gov (United States)

    Castillo Cádiz, Octavio; Villasenín Parrado, Lorena; Borgna Christie, Vincenzo; Gallegos Méndez, Iván; Martínez Corta, Virginia

    2016-06-20

    Bacille Calmette-Guerin intravesical treatment is the most effective treatment for reducing the recurrence of non-muscle-invasive urothelial carcinomas. This treatment can sometimes have side effects and serious complications. Granulomatous prostatitis is a common histological finding but it rarely has a clinical presentation. We report a case of a 75-year-old, type 2 diabetic, male patient who was diagnosed with urothelial in situ carcinoma, for which he began treatment with Bacille Calmette-Guerin instillations. Five years later the patient presented nocturia, pollakiuria, severe urgency, and intense and recurrent perineal pain associated with marked elevation of prostatic specific antigen. A prostatic biopsy was performed that showed a moderate to severe granulomatous prostatitis related to bacille Calmette-Guerin. The patient received full antituberculosis combination drugs with a favorable clinical response.

  12. Granulomas do pênis: uma complicação rara da terapia intravesical com Bacilo Calmette-Guérin Granulomas of the penis: a rare complication of intravesical therapy with Bacillus Calmette-Guerin

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    Sara Isabel Alcântara Lestre

    2011-08-01

    Full Text Available A imunoterapia com o Bacilo Calmette-Guérin é amplamente usada no tratamento e profilaxia da neoplasia urotelial superficial. As complicações associadas ao tratamento são comuns. Os autores relatam um caso de inflamação granulomatosa do pênis, associada à terapia intravesical com Bacilo Calmette-Guérin, com múltiplos nódulos eritematosos indolores localizados na glande. É também efetuada uma revisão da literatura. A balanopostite granulomatosa é uma complicação rara associada à imunoterapia com Bacilo Calmette-Guérin, com uma apresentação clinicamente heterogênea que pode dificultar o diagnóstico. O seu reconhecimento clínico é essencial para o início precoce de tuberculostáticos e interrupção de Bacilo Calmette-GuérinImmunotherapy with Bacillus Calmette-Guérin is widely used for treatment and prophylaxis of superficial urothelial cancer. Complications associated with Bacillus Calmette-Guérin treatment are common. The authors describe a case of granulomatous inflammation of the penis associated with intravesical Bacillus Calmette-Guérin therapy, presenting with multiple erythematous and painless nodules located on the glans. A review of the literature is also performed. Granulomatous balanoposthitis is a rare complication of Bacillus Calmette-Guérin immunotherapy, with heterogeneous clinical presentation, which can make the diagnosis difficult. Its clinical recognition is essential for early start of therapy with antitubercular agents and interruption of Bacillus Calmette-Guérin

  13. Disseminated Bacille Calmette-Guerin (BCG) disease in an infant with severe combined immunodeficiency.

    Science.gov (United States)

    Sohail, Shagufta; Afzal, Muhammad; Anwar, Vaqas; Shama, Quratulain

    2014-11-01

    Bacille Calmette-Guerin (BCG) vaccine is administered to all newborns in countries where tuberculosis is still endemic. It is a live attenuated vaccine and considered quite safe in immunocompetent children. Disseminated BCG disease is the most serious complication seen only in individuals with underlying primary or secondary immunodeficiencies. We report a case of disseminated BCG disease in an infant with Severe Combined Immunodeficiency (SCID) who received BCG administration prior to diagnosis of SCID.

  14. Late-onset granulomatous prostatitis following intravesical bacille Calmette-Guerin therapy: case report

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    Octavio Castillo Cádiz

    2016-06-01

    Full Text Available Resumen El bacilo de Calmette-Guerin es el tratamiento intravesical más efectivo para disminuir la recurrencia de los carcinomas uroteliales no-músculo-invasivos. La aplicación de este tratamiento en ocasiones puede presentar efectos secundarios y, excepcionalmente, complicaciones graves. La prostatitis granulomatosa es un hallazgo histológico frecuente pero una entidad rara desde el punto de vista clínico. Se presenta el caso de un paciente de 75 años, diabético tipo 2, que fue diagnosticado de carcinoma in situ vesical, para lo cual inició tratamiento con bacilo de Calmette-Guerin intravesical. El paciente consultó cinco años después por presentar cuadro de nicturia, frecuencia miccional aumentada, urgencia miccional grave y dolor perineal intenso y recurrente asociado a una curva de antígeno prostático específico con marcada elevación. Se le realizó biopsia prostática que mostró una prostatitis crónica granulomatosa de grado moderado a grave relacionada a bacilo de Calmette-Guerin. El paciente recibió esquema antituberculoso completo con buena respuesta clínica.

  15. 卡介菌多糖核酸与化疗药物联合治疗肺癌的实验研究%A Experimental Study on Treatment of Lung Cancer with Bacillus Calmette-Guerin Polysaccharides Nucleic Acid Combined with Chemotherapy

    Institute of Scientific and Technical Information of China (English)

    刘建新; 金龙玉; 洪朝; 黄碚; 周永生; 朱兵; 刘科颍

    2001-01-01

    Objective To investigate the mechanism of treatment of lung cancer with Bacillus Calmette-Guerin Polysaccharides nucleic acid (BCG-PSN) combined with Cyclophosphamide(Cy). Methods 130 mice were divided into three groups (A,B and C groups). Gruop A was further divided into four subgroups . Subgroup A-1 was given normal saline orally 0.25mg/10g*d-1; Subgroup A-2 was given levomisole PO. 0.25mg/10g*d-1; Subgroup A-3 was given BCG-PSN PO.0.325mg/10g*d-1 and subgroup A-4 was given BCG-PSN by intra-abdominal injection 0.25mg/10g*d-1. Group B was given identical drugs with group A, after fifth day given 20% sheep red blood cell 0.2ml by intra-abdominal injection for five days . Gruop C divided into five subgroups;Subgroup C1-4 were given same drugs with group A,then given 0.4% Cy by subcutaneous injection 0.25mg/10g*d-1 from fifth day to tenth day.Subgroup C-5 was normal contral group, was given normal saline 0.25mg/10g*d-1 PO and subcutaneous injection 0.25mg/10g*d-1.Results Oral administration alone or intra-abdominal injection in subgroup A-4 obviously enhanced the phagocytosis of macrophages.The subcutaneous injection of Cy could obviously decrease anti-tumour immunity, including activity of interleukin-2 (IL-2),natural killer(NK) cell, lyphocyte trasformation, and macrophages phagocytosis. BCG-PSN could increase and rehabilitate above anti-tumour immunological function. Conclusions Combinative treatment of BCG-PSN with Cy could remarkable enhance the immunocompetence.This experiment may provide a theoretical evidence for BCG-PSN combined with chemotheapeutic agents applied to treat the lung cancer.%目的探讨卡介菌多糖核酸(BCG-PSN)与化疗药物联合治疗肺癌的机理。方法小鼠130只,分为A、B、C三组,分别测定单纯口服或腹腔内注射BCG-PSN后小鼠的巨噬细胞吞噬功能、淋巴细胞转化刺激指数及溶血素试验;单纯皮下注射环磷酰胺(Cy)和BCG-PSN与Cy联合用药后小鼠的自然

  16. Sternal osteomyelitis after bacillus Calmette-Guérin vaccination

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    Rolandas Selvestravičius

    2016-09-01

    Full Text Available Presented here is the case of a nine-month-old boy with the osteomyelitis of the upper area sternum caused by bacillus Calmette-Guerin (BCG, the Danish 1331 strain vaccine against tuberculosis. Upon examination, a swelling of approximately 2×3 cm diameter was observed in the upper sternal area. The mass was hard, fixed and sensitive to palpation with no local skin hyperaemia. Chest X-rays revealed a round mass anterior to the sternum, suggesting a diagnosis of osteomyelitis. A consequent sternal biopsy was performed and Mycobacterium bovis BCG was identified by a positive growth culture.

  17. Sternal Osteomyelitis after Bacillus Calmette-Guérin Vaccination

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    Selvestravičius, Rolandas; Sučilienė, Elena; Saniukas, Kęstutis; Bobelytė, Odeta; Usonis, Vytautas

    2016-01-01

    Presented here is the case of a nine-month-old boy with the osteomyelitis of the upper area sternum caused by bacillus Calmette-Guerin (BCG), the Danish 1331 strain vaccine against tuberculosis. Upon examination, a swelling of approximately 2×3 cm diameter was observed in the upper sternal area. The mass was hard, fixed and sensitive to palpation with no local skin hyperaemia. Chest X-rays revealed a round mass anterior to the sternum, suggesting a diagnosis of osteomyelitis. A consequent sternal biopsy was performed and Mycobacterium bovis BCG was identified by a positive growth culture. PMID:27777704

  18. Peripheral blood and marrow findings in disseminated bacille Calmette-Guerin infection.

    Science.gov (United States)

    Kumar, Perikala Vijayananda; Monabati, Ahmad; Kadivar, Rahim; Soleimanpour, Hossein

    2005-02-01

    The authors describe an unusual case of a disseminated bacille Calmette-Guerin (BCG) infection in a 3-month-old girl who presented with a huge hepatosplenomegaly, fever, and pancytopenia. Clinically, an infantile kala-azar or lymphoma/leukemia was suspected. However, after thorough clinical and paraclinical investigations, the case was diagnosed as a disseminated BCG infection. The child died 2 weeks after starting antituberculosis treatment. Autopsy revealed diffuse histiocytic infiltration in the liver, spleen, and mesenteric lymph nodes, which were loaded with acid-fast bacilli. Three interesting findings were noticed in this case: circulating monocytes in the peripheral blood were loaded with ghost acid-fast bacilli; bone marrow smears revealed numerous Gaucher cell-like macrophages loaded with negative images of Mycobacterium tuberculi; and there was extensive marrow necrosis. These findings have not been previously reported in the literature.

  19. Bilateral symmetrical corneal melting following intravesical Bacille Calmette-Guerin therapy for bladder carcinoma

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    Chandana Chakraborty

    2012-01-01

    Full Text Available A 63-year-old man with unremarkable previous ocular history presented with bilateral symmetrical corneal ulceration along with mucopurulent conjunctivitis and dry eye 10 days after the fourth dose of intravesical Bacille Calmette-Guerin (BCG instillation for treatment of bladder carcinoma. Slit lamp examination revealed thinning of the cornea at the base of the ulcer in both eyes. Conjunctival swab and scraping from ulcer sent for Gram and acid fast bacilli stain and culture were negative. On the basis of history, clinical examination, and laboratory investigations, we diagnosed it as bilateral immune mediated sterile corneal ulceration along with mucopurulent conjunctivitis and dry eye. He was treated with topical antibiotics, cycloplegics, cyclosporine, lubricant gel, and bandage contact lens. There was progressive stromal melting, descemetocele formation, and perforation in the inferior part of cornea in both the eyes. He was treated with pulse steroid and paramedian tarsorraphy in both eyes. The patient was subsequently lost to follow-up. We report this case to highlight this rare complication of BCG therapy, in order to improve their management protocol in patients with similar clinical profile. We could not find a similar case after thorough PubMed search.

  20. [Generalized BCG infection after intravesical instillations of Calmette-Guerin bacillus].

    Science.gov (United States)

    de Saint Martin, L; Boiron, C; Poveda, J D; Herreman, G

    1993-10-02

    BCG has been disappointing as immunotherapy of numerous cancers, but it has been clinically successful in the intravesical treatment of bladder carcinomas sparing the muscle coat; it has indeed become the reference treatment for this type of cancer. However, complications are repeatedly reported, including generalized BCGitis. We report such a case with positive BCG culture. From the cases already published there emerges a homogeneous and often subacute clinical presentation suggestive of an ordinary pathogen. Bacteriology is not very helpful, even when recent techniques are used, and therefore the diagnosis rests on the context and, when samples are taken, on suggestive histological findings. To discuss the physiopathology of BCGitis--generalized immune reaction or multifocal BCG proliferation--is not useless since treatment depends on it. It is probable that these 2 mechanisms working together can be incriminated justifying the prescription of both antibiotics and corticosteroids. When this is done, the prognosis seems to be favourable in most patients. Yet a strict respect of contra-indications and a very careful subsequent radiotherapy should reduce the risks.

  1. Molecular profiling of Mycobacterium tuberculosis identifies tuberculosinyl nucleoside products of the virulence-associated enzyme Rv3378c

    NARCIS (Netherlands)

    Layre, Emilie; Lee, Ho Jun; Young, David C.; Martinot, Amanda Jezek; Buter, Jeffrey; Minnaard, Adriaan J.; Annand, John W.; Fortune, Sarah M.; Snider, Barry B.; Matsunaga, Isamu; Rubin, Eric J.; Alber, Tom; Moody, D. Branch

    2014-01-01

    To identify lipids with roles in tuberculosis disease, we systematically compared the lipid content of virulent Mycobacterium tuberculosis with the attenuated vaccine strain Mycobacterium bovis bacillus Calmette-Guerin. Comparative lipidomics analysis identified more than 1,000 molecular differences

  2. Protection Induced by Simultaneous Subcutaneous and Endobronchial Vaccination with BCG/BCG and BCG/Adenovirus Expressing Antigen 85A against Mycobacterium bovis in Cattle.

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    Gillian S Dean

    Full Text Available The incidence of bovine tuberculosis (bTB in the GB has been increasing since the 1980s. Immunisation, alongside current control measures, has been proposed as a sustainable measure to control bTB. Immunisation with Mycobacterium bovis bacillus Calmette-Guerin (BCG has been shown to protect against bTB. Furthermore, much experimental data indicates that pulmonary local immunity is important for protection against respiratory infections including Mycobacterium tuberculosis and that pulmonary immunisation is highly effective. Here, we evaluated protection against M. bovis, the main causative agent of bTB, conferred by BCG delivered subcutaneously, endobronchially or by the new strategy of simultaneous immunisation by both routes. We also tested simultaneous subcutaneous immunisation with BCG and endobronchial delivery of a recombinant type 5 adenovirus expressing mycobacterial antigen 85A. There was significantly reduced visible pathology in animals receiving the simultaneous BCG/BCG or BCG/Ad85 treatment compared to naïve controls. Furthermore, there were significantly fewer advanced microscopic granulomata in animals receiving BCG/Ad85A compared to naive controls. Thus, combining local and systemic immunisation limits the development of pathology, which in turn could decrease bTB transmission.

  3. Cell wall lipids from Mycobacterium bovis BCG are inflammatory when inoculated within a gel matrix: characterization of a new model of the granulomatous response to mycobacterial components.

    Science.gov (United States)

    Rhoades, Elizabeth R; Geisel, Rachel E; Butcher, Barbara A; McDonough, Sean; Russell, David G

    2005-05-01

    The chronic inflammatory response to Mycobacterium generates complex granulomatous lesions that balance containment with destruction of infected tissues. To study the contributing factors from host and pathogen, we developed a model wherein defined mycobacterial components and leukocytes are delivered in a gel, eliciting a localized response that can be retrieved and analysed. We validated the model by comparing responses to the cell wall lipids from Mycobacterium bovis bacillus Calmette-Guerin (BCG) to reported activities in other models. BCG lipid-coated beads and bone marrow-derived macrophages (input macrophages) were injected intraperitoneally into BALB/c mice. Input macrophages and recruited peritoneal exudate cells took up fluorescently tagged BCG lipids, and matrix-associated macrophages and neutrophils produced tumor necrosis factor, interleukin-1alpha, and interleukin-6. Leukocyte numbers and cytokine levels were greater in BCG lipid-bearing matrices than matrices containing non-coated or phosphatidylglycerol-coated beads. Leukocytes arrived in successive waves of neutrophils, macrophages and eosinophils, followed by NK and T cells (CD4(+), CD8(+), or gammadelta) at 7 days and B cells within 12 days. BCG lipids also predisposed matrices for adherence and vascularization, enhancing cellular recruitment. We submit that the matrix model presents pertinent features of the murine granulomatous response that will prove to be an adaptable method for study of this complex response.

  4. A polymerase chain reaction and enzyme linked immunosorbent assay based approach for diagnosis and differentiation between vaccinated and infected cattle with Mycobacterium bovis

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    Mohamed Sabry

    2014-01-01

    Full Text Available Background: In most African and Arabic countries tuberculosis (TB causes great economic losses in bovine species and constitutes serious zoonotic problem. As the traditional diagnostic method delay the research because of low sensitivity and specificity, a rapid method of diagnosis is of outmost importance. Aim: The study was designed to evaluate the two rapid diagnostic methods of TB in cattle, further to differentiate between infected and bacillus Calmette-Guerin (BCG vaccinated animals. Materials and Methods: Intradermal tuberculin test was applied to 300 cattle. Of these cattle, 15 cattle were vaccinated from cattle negative to tuberculin test with BCG. Blood samples were taken for lymphocyte separation to apply polymerase chain reaction (PCR upon and for serum preparation for the enzyme-linked immunosorbent assay (ELISA application, this blood collected from 65 cattle classified into three groups, viz. positive tuberculin test (35 animals, negative tuberculin test (15 animals, and vaccinated cow with BCG (15 animals. From blood samples lymphocytes were separated and the isolated lymphocytes were subjected to PCR and serum for ELISA application. Blood samples, specimens from lymph nodes and specific tissues were taken for PCR and for cultivation and isolation of Mycobacterium bovis. Results and Conclusions: The results of this study revealed that PCR can be used as rapid efficient and accurate diagnostic test in detection of ruminant TB. Moreover, cattle′s ELISA reading showed higher sensitivity in positive tuberculin animals. However, the differentiations between vaccinated and infected animals not clear by using a single antigen only.

  5. Increased B and T Cell Responses in M. bovis Bacille Calmette-Guerin Vaccinated Pigs Co-Immunized with Plasmid DNA Encoding a Prototype Tuberculosis Antigen.

    Directory of Open Access Journals (Sweden)

    Nicolas Bruffaerts

    Full Text Available The only tuberculosis vaccine currently available, bacille Calmette-Guérin (BCG is a poor inducer of CD8(+ T cells, which are particularly important for the control of latent tuberculosis and protection against reactivation. As the induction of strong CD8(+ T cell responses is a hallmark of DNA vaccines, a combination of BCG with plasmid DNA encoding a prototype TB antigen (Ag85A was tested. As an alternative animal model, pigs were primed with BCG mixed with empty vector or codon-optimized pAg85A by the intradermal route and boosted with plasmid delivered by intramuscular electroporation. Control pigs received unformulated BCG. The BCG-pAg85A combination stimulated robust and sustained Ag85A specific antibody, lymphoproliferative, IL-6, IL-10 and IFN-γ responses. IgG1/IgG2 antibody isotype ratio reflected the Th1 helper type biased response. T lymphocyte responses against purified protein derivative of tuberculin (PPD were induced in all (BCG vaccinated animals, but responses were much stronger in BCG-pAg85A vaccinated pigs. Finally, Ag85A-specific IFN-γ producing CD8(+ T cells were detected by intracellular cytokine staining and a synthetic peptide, spanning Ag85A131-150 and encompassing two regions with strong predicted SLA-1*0401/SLA-1*0801 binding affinity, was promiscuously recognized by 6/6 animals vaccinated with the BCG-pAg85A combination. Our study provides a proof of concept in a large mammalian species, for a new Th1 and CD8(+ targeting tuberculosis vaccine, based on BCG-plasmid DNA co-administration.

  6. Adaptive immunity in the colostrum-deprived calf: Response to early vaccination with Mycobacterium bovis, strain Bacille Calmette Guerin (BCG) and ovalbumin

    Science.gov (United States)

    Responses of the newborn calf to vaccination are variable and frequently characterized by marginal antibody (Ab) responses. The present study evaluated effects of colostrum ingestion on the adaptive immune response of the preruminant calf to early vaccination. Colostrum-fed (CF) and colostrum-depriv...

  7. Prognostic value of Bcl-2 and Bax tumor cell expression in patients with non muscle-invasive bladder cancer receiving bacillus Calmette-Guerin immunotherapy.

    Science.gov (United States)

    Ajili, Faouzia; Kaabi, Belhassen; Darouiche, Amine; Tounsi, Haifa; Kourda, Nadia; Chebil, Mohamed; Manai, Mohamed; Boubaker, Samir

    2012-02-01

    Apoptosis is the distinctive form of programmed cell death that complements cell proliferation in maintaining normal tissue homeostasis. The significance of constitutive apoptosis in the recurrence of Non Muscle Invasive Bladder Cancer has yet to be investigated. The aim of this study is to investigate the prognostic significance of Bax and Bcl-2 in terms of recurrence after BCG immunotherapy. Immunohistochemical analysis was performed on frozen biopsies to evaluate bcl-2 and Bax proteins expression in 28 cases of NMIBC. All patients with confirmed NMIBC were treated with intravesical BCG-immunotherapy. The follow up was performed for 26 months. The correlation between clinicopathological, immunohistochemical data and the response to BCG therapy was performed. Univariate analysis showed that, PT1 stage, High grade and Bax expression increased significantly the risk of recurrence (P = 0.015, P = 0.015 and P= 0.034 respectively). In addition, multivariate analysis selected the model involving stage, age, Bax and Bcl-2 expression as the best independent variables of recurrence. In conclusion, the expression of Bcl-2 and Bax in NMIBC could have a prognostic value in assessing the risk of recurrence after BCG immunotherapy. These findings require further investigations on larger cohort in order to ascertain new molecular markers of the response to BCG immunotherapy.

  8. Stable expression of Mycobacterium bovis antigen 85B in auxotrophic M. bovis bacillus Calmette-Guérin

    Science.gov (United States)

    Rizzi, Caroline; Peiter, Ana Carolina; Oliveira, Thaís Larré; Seixas, Amilton Clair Pinto; Leal, Karen Silva; Hartwig, Daiane Drawanz; Seixas, Fabiana Kommling; Borsuk, Sibele; Dellagostin, Odir Antônio

    2017-01-01

    BACKGROUND Bovine tuberculosis (TB) is a zoonotic disease caused by Mycobacterium bovis, responsible for causing major losses in livestock. A cost effective alternative to control the disease could be herd vaccination. The bacillus Calmette-Guérin (BCG) vaccine has a limited efficacy against bovine TB, but can improved by over-expression of protective antigens. The M. bovis antigen 85B demonstrates ability to induce protective immune response against bovine TB in animal models. However, current systems for the construction of recombinant BCG expressing multiple copies of the gene result in strains of low genetic stability that rapidly lose the plasmid in vivo. Employing antibiotic resistance as selective markers, these systems also compromise vaccine safety. We previously reported the construction of a stable BCG expression system using auxotrophic complementation as a selectable marker. OBJECTIVES The fundamental aim of this study was to construct strains of M. bovis BCG Pasteur and the auxotrophic M. bovis BCG ΔleuD expressing Ag85B and determine their stability in vivo. METHODS Employing the auxotrophic system, we constructed rBCG strains that expressed M. bovis Ag85B and compared their stability with a conventional BCG strain in mice. Stability was measured in terms of bacterial growth on the selective medium and retention of antigen expression. FINDINGS The auxotrophic complementation system was highly stable after 18 weeks, even during in vivo growth, as the selective pressure and expression of antigen were maintained comparing to the conventional vector. MAIN CONCLUSION The Ag85B continuous expression within the host may generate a stronger and long-lasting immune response compared to conventional systems. PMID:28177046

  9. Vaccine approaches for bovine tuberculosis: Correlates of protection and relevance to human tuberculosis

    Science.gov (United States)

    Tuberculosis (TB), primarily due to Mycobacterium tuberculosis in humans and Mycobacterium bovis in cattle, is a classic model of the One Health Concept. M. bovis Bacillus Calmette Guerin (BCG) was first proven effective in cattle prior to use in humans. Recent experimental trials with cattle have d...

  10. Role of Phosphatidylinositol Mannosides in the Interaction between Mycobacteria and DC-SIGN

    NARCIS (Netherlands)

    Driessen, N.N.; Ummels, R.; Maaskant, J.J.; Gurcha, S.S.; Besra, G.S.; Ainge, G.D.; Larsen, D.S.; Painter, G.F.; Vandenbroucke-Grauls, C.M.J.E.; Geurtsen, J.J.G.; Appelmelk, B.J.

    2009-01-01

    2)-linked mannosyl residues identical to the mannose cap on ManLAM, but not for di- and tetramannosylated PIM2 and PIM4, respectively. To determine the role of PIM6 in the binding of whole mycobacteria to DC-SIGN, a mutant strain of M. bovis bacillus Calmette-Guerin deficient in the production of PI

  11. Recombinant Adenovirus Delivery of Calreticulin-ESAT-6 Produces an Antigen-Specific Immune Response but no Protection Against a Mycobacterium Tuberculosis Challenge

    NARCIS (Netherlands)

    Esparza-Gonzalez, S. C.; Troy, A.; Troudt, J.; Loera-Arias, M. J.; Villatoro Hernandez, Julio; Torres-Lopez, E.; Ancer-Rodriguez, J.; Gutierrez-Puente, Y.; Munoz-Maldonado, G.; Saucedo-Cardenas, O.; Montes-de-Oca-Luna, R.; Izzo, A.

    2012-01-01

    Bacillus CalmetteGuerin (BCG) has failed to efficaciously control the worldwide spread of the disease. New vaccine development targets virulence antigens of Mycobacterium tuberculosis that are deleted in Mycobacterium bovis BCG. Immunization with ESAT-6 and CFP10 provides protection against M. tuber

  12. BCG (Bacille Calmette-Guerin) Vaccine

    Science.gov (United States)

    ... the Facts Tuberculosis - The Connection between TB and HIV 12-Dose Regimen for Latent TB Infection-Patient Education Brochure Posters Mantoux Tuberculin Skin Test Wall Chart World TB Day Think TB Stop TB Reports & Articles Morbidity and Mortality Weekly Reports (MMWRs) DTBE Authored ...

  13. Mycobacterium bovis Bacillus Calmette-Guérin-Induced Macrophage Cytotoxicity against Bladder Cancer Cells

    Directory of Open Access Journals (Sweden)

    Yi Luo

    2010-01-01

    Full Text Available Many details of the molecular and cellular mechanisms involved in Mycobacterium bovis bacillus Calmette-Guérin (BCG immunotherapy of bladder cancer have been discovered in the past decades. However, information on a potential role for macrophage cytotoxicity as an effector mechanism is limited. Macrophages play pivotal roles in the host innate immunity and serve as a first line of defense in mycobacterial infection. In addition to their function as professional antigen-presenting cells, the tumoricidal activity of macrophages has also been studied with considerable interest. Studies have shown that activated macrophages are potent in killing malignant cells of various tissue origins. This review summarizes the current understanding of the BCG-induced macrophage cytotoxicity toward bladder cancer cells with an intention to inspire investigation on this important but underdeveloped research field.

  14. Proteomic profile of culture filtrate from the Brazilian vaccine strain Mycobacterium bovis BCG Moreau compared to M. bovis BCG Pasteur

    Directory of Open Access Journals (Sweden)

    Degrave Wim M

    2011-04-01

    Full Text Available Abstract Background Bacille Calmette-Guerin (BCG is currently the only available vaccine against tuberculosis (TB and comprises a heterogeneous family of sub-strains with genotypic and phenotypic differences. The World Health Organization (WHO affirms that the characterization of BCG sub-strains, both on genomic and proteomic levels, is crucial for a better comprehension of the vaccine. In addition, these studies can contribute in the development of a more efficient vaccine against TB. Here, we combine two-dimensional electrophoresis (2DE and mass spectrometry to analyse the proteomic profile of culture filtrate proteins (CFPs from M. bovis BCG Moreau, the Brazilian vaccine strain, comparing it to that of BCG Pasteur. CFPs are considered of great importance given their dominant immunogenicity and role in pathogenesis, being available for interaction with host cells since early infection. Results The 2DE proteomic map of M. bovis BCG Moreau CFPs in the pH range 3 - 8 allowed the identification of 158 spots corresponding to 101 different proteins, identified by MS/MS. Comparison to BCG Pasteur highlights the great similarity between these BCG strains. However, quantitative analysis shows a higher expression of immunogenic proteins such as Rv1860 (BCG1896, Apa, Rv1926c (BCG1965c, Mpb63 and Rv1886c (BCG1923c, Ag85B in BCG Moreau when compared to BCG Pasteur, while some heat shock proteins, such as Rv0440 (BCG0479, GroEL2 and Rv0350 (BCG0389, DnaK, show the opposite pattern. Conclusions Here we report the detailed 2DE profile of CFPs from M. bovis BCG Moreau and its comparison to BCG Pasteur, identifying differences that may provide relevant information on vaccine efficacy. These findings contribute to the detailed characterization of the Brazilian vaccine strain against TB, revealing aspects that may lead to a better understanding of the factors leading to BCG's variable protective efficacy against TB.

  15. Effects of berberine hydrochloride on CYP450 total content and expression in BCG-induced immune hepatic injury in mice and its possible mechanism

    Institute of Scientific and Technical Information of China (English)

    XinWANG; DanLI; Jun-jieZHANG; Xiu-yunBU; Guo-liangZHANG

    2004-01-01

    AIM:To investigate effects of berberine hydrochloride on hepatic cytochrome P450 in BCG-induced immunological hepatic injury in BALB/c mice and its possible mechanism. METHODS: Immunological liver injury was induced by intravenous injection of Mycobacterium bovis bacillus Calmette-Guerin (BCG, 125 mg/kg) in BALB/c mice. After one week stimulated by BCG,berberine hydrochloride (10, 25, 50, 75, and 100 mg/kg,respectively, qid 7 d) was administrated by intragastric

  16. Cloning and expression of the fusion protein of interleukin-2 and ESAT6 in Mycobacterium bovis Bacillus Calmette Guérin strain

    Institute of Scientific and Technical Information of China (English)

    FAN Xiong-lin; WANG Li-mei; LU Xian-yu; TU Zhi-guang; SHI Chang-hong; XU Zhi-kai

    2005-01-01

    @@ Tuberculosis (TB) is the most common cause of death in infectious diseases; it is estimated that approximately 2 million people per year die of TB. The present available TB vaccine is a live attenuated strain, Mycobacterium bovis Bacillus Calmette Guérin (BCG). However, it has been shown that BCG has variable protective efficacy, ranging from 0 to 85% in different clinical experiments.1 Therefore, a new TB vaccine is urgently needed.

  17. Human T cell responses induced by vaccination with Mycobacterium bovis bacillus Calmette-Guérin

    DEFF Research Database (Denmark)

    Ravn, P; Boesen, H; Pedersen, B K;

    1997-01-01

    have studied in vitro cell-mediated immune responses primed by BCG vaccination in 22 healthy Danish donors with different levels of in vitro purified protein derivative (PPD) reactivity before vaccination. The study demonstrated a markedly different development of reactivity to mycobacterial Ags......Many aspects of the widely used bacillus Calmette-Guérin (BCG) vaccine against tuberculosis are still the subject of controversy. There is a huge variation in efficacy from one clinical trial to another and no relationship between vaccine-induced skin test conversion and subsequent protection. We...... depending on the prevaccination sensitivity to PPD. Previously sensitized donors mounted a potent and highly accelerated recall response within the first week of BCG vaccination. Nonsensitized donors, in contrast, exhibited a gradually increasing responsiveness to mycobacterial Ags, reaching maximal levels...

  18. A New Screen for Tuberculosis Drug Candidates Utilizing a Luciferase-Expressing Recombinant Mycobacterium bovis Bacillus Calmette-Gueren.

    Directory of Open Access Journals (Sweden)

    Yuriko Ozeki

    Full Text Available Tuberculosis (TB is a serious infectious disease caused by a bacterial pathogen. Mortality from tuberculosis was estimated at 1.5 million deaths worldwide in 2013. Development of new TB drugs is needed to not only to shorten the medication period but also to treat multi-drug resistant and extensively drug-resistant TB. Mycobacterium tuberculosis (Mtb grows slowly and only multiplies once or twice per day. Therefore, conventional drug screening takes more than 3 weeks. Additionally, a biosafety level-3 (BSL-3 facility is required. Thus, we developed a new screening method to identify TB drug candidates by utilizing luciferase-expressing recombinant Mycobacterium bovis bacillus Calmette-Guéren (rBCG. Using this method, we identified several candidates in 4 days in a non-BSL-3 facility. We screened 10,080 individual crude extracts derived from Actinomyces and Streptomyces and identified 137 extracts which possessed suppressive activity to the luciferase of rBCG. Among them, 41 compounds inhibited the growth of both Mtb H37Rv and the extensively drug-resistant Mtb (XDR-Mtb strains. We purified the active substance of the 1904-1 extract, which possessed strong activity toward rBCG, Mtb H37Rv, and XDR-Mtb but was harmless to the host eukaryotic cells. The MIC of this substance was 0.13 μg/ml, 0.5 μg/ml, and 2.0-7.5 μg/ml against rBCG, H37Rv, and 2 XDR-strains, respectively. Its efficacy was specific to acid-fast bacterium except for the Mycobacterium avium intracellular complex. Mass spectrometry and nuclear magnetic resonance analyses revealed that the active substance of 1904-1 was cyclomarin A. To confirm the mode of action of the 1904-1-derived compound, resistant BCG clones were used. Whole genome DNA sequence analysis showed that these clones contained a mutation in the clpc gene which encodes caseinolytic protein, an essential component of an ATP-dependent proteinase, and the likely target of the active substance of 1904-1. Our method

  19. 吉安市城乡儿童卡介菌纯蛋白衍生物试验结果分析%Analysis on the test results of calmette- guerin bacillus ppd among the children in urban and rural of Jian

    Institute of Scientific and Technical Information of China (English)

    黄光明; 刘晓东; 罗智勇

    2011-01-01

    目的 了解卡介苗的接种质量,分析影响因素,为改善预防接种工作提供依据.方法 应用卡介菌纯蛋白衍生物( BCG- PPD)试验方法,随机抽取城市和乡村各250名3个月~1岁儿童,接种卡介苗3个月后进行PPD阳转率调查.结果BCG- PPD试验总阳性率88.60%,无性别差异,城市儿童BCG- PPD试验阳性率高于农村儿童.BCG-PPD试验阳性率与卡疤大小呈正相关.结论 我市儿童卡介苗接种质量较高,应改善农村预防接种条件,加强对BCG预防接种人员特别是农村预防接种人员的技术培训,有利于提高预防接种质量.%Objective To understand the quality of BCG vaccination, analysis influence factors, and provide basis for improving the vaccination work. Method Application of BCG purified protein derivative ( BCG - PPD ) test method, random selected 250 children of 3 months to 1 years old among each city and countryside, after 3 months, investigation the ppd seroconversion rate. Results The total positive rate of BCG - PPD test was 88. 60% , no sex differences, the positive rate of city children were higher than rural children. BCG - PPD test positive rate and card scar size showed positive correlation. Conclusions The BCG vaccination quality was high in our city, but we should improve rural preventive inoculation conditions, strengthen technical training of BCG vaccination staff especially rural vaccination staff, it profit for improving the quality of vaccination.

  20. Study on Effect of Bacillus Calmette-Guerin Combined with Levamisole in Treating Children with Recurrent Respiratory Tract Infections%卡介苗素联合左旋咪唑治疗小儿反复呼吸道感染的疗效研究

    Institute of Scientific and Technical Information of China (English)

    陈晓玲; 王宇; 王玉玲

    2011-01-01

    目的 探讨卡介苗素联合左旋咪唑治疗小儿反复呼吸道感染(RRTI)的临床效果.方法 将2008年1月~2009年1月间在我院住院治疗的RRTI患儿随机分为治疗组和对照组,两组均给予常规治疗,治疗组同时给予卡介苗素联合左旋咪唑治疗.结果 治疗组CD3+、CD4+以及CD4+/CD8+明显高于对照组,两组比较差异具有统计学意义(P<0.05);治疗组在呼吸道感染次数、发热时间、咳嗽时间、抗生素使用时间方面明显优于对照组,两者分别比较差异具有统计学意义(P<0.05);治疗组总有效率为95.00%.对照组为65.00%,两者比较差异具有统计学意义(P<0.05);两组患儿均未见明显的治疗不良反应.结论 卡介苗素联合左旋咪唑治疗小儿反复呼吸道感染的临床效果确切,无毒副作用,值得推荐应用.

  1. Different Transcriptional Profiles of Human Monocyte-Derived Dendritic Cells Infected with Distinct Strains of Mycobacterium tuberculosis and Mycobacterium bovis Bacillus Calmette-Guérin

    Directory of Open Access Journals (Sweden)

    Nunzia Sanarico

    2011-01-01

    Full Text Available In order to analyze dendritic cells (DCs activation following infection with different mycobacterial strains, we studied the expression profiles of 165 genes of human monocyte-derived DCs infected with H37Rv, a virulent Mycobacterium tuberculosis (MTB laboratory strain, CMT97, a clinical MTB isolate, Mycobacterium bovis bacillus Calmette-Guérin (BCG, Aventis Pasteur, and BCG Japan, both employed as vaccine against tuberculosis. The analysis of the gene expression reveals that, despite a set of genes similarly modulated, DCs response resulted strain dependent. In particular, H37Rv significantly upregulated EBI3 expression compared with BCG Japan, while it was the only strain that failed to release a significant IL-10 amount. Of note, BCG Japan showed a marked increase in CCR7 and TNF-α expression regarding both MTB strains and it resulted the only strain failing in exponential intracellular growth. Our results suggest that DCs display the ability to elicit a tailored strain-specific immune response.

  2. Improving Mycobacterium bovis Bacillus Calmette-Guèrin as a Vaccine Delivery Vector for Viral Antigens by Incorporation of Glycolipid Activators of NKT Cells

    Science.gov (United States)

    Kharkwal, Shalu S.; Carreño, Leandro J.; Johnson, Alison J.; Kunnath-Velayudhan, Shajo; Liu, Zheng; Bittman, Robert; Jervis, Peter J.; Cox, Liam R.; Besra, Gurdyal S.; Wen, Xiangshu; Yuan, Weiming; Tsuji, Moriya; Li, Xiangming; Ho, David D.; Chan, John; Lee, Sunhee; Frothingham, Richard; Haynes, Barton F.; Panas, Michael W.; Gillard, Geoffrey O.; Sixsmith, Jaimie D.; Korioth-Schmitz, Birgit; Schmitz, Joern E.; Larsen, Michelle H.; Jacobs, William R.; Porcelli, Steven A.

    2014-01-01

    Recombinant Mycobacterium bovis bacillus Calmette-Guèrin (rBCG) has been explored as a vector for vaccines against HIV because of its ability to induce long lasting humoral and cell mediated immune responses. To maximize the potential for rBCG vaccines to induce effective immunity against HIV, various strategies are being employed to improve its ability to prime CD8+ T cells, which play an important role in the control of HIV infections. In this study we adopted a previously described approach of incorporating glycolipids that activate CD1d-restricted natural killer T (NKT) cells to enhance priming of CD8+ T cells by rBCG strains expressing an SIV Gag antigen (rBCG-SIV gag). We found that the incorporation of the synthetic NKT activating glycolipid α-galactosylceramide (α-GC) into rBCG-SIV gag significantly enhanced CD8+ T cell responses against an immunodominant Gag epitope, compared to responses primed by unmodified rBCG-SIV gag. The abilities of structural analogues of α-GC to enhance CD8+ T cell responses to rBCG were compared in both wild type and partially humanized mice that express human CD1d molecules in place of mouse CD1d. These studies identified an α-GC analogue known as 7DW8-5, which has previously been used successfully as an adjuvant in non-human primates, as a promising compound for enhancing immunogenicity of antigens delivered by rBCG.vectors. Our findings support the incorporation of synthetic glycolipid activators of NKT cells as a novel approach to enhance the immunogenicity of rBCG-vectored antigens for induction of CD8+ T cell responses. The glycolipid adjuvant 7DW8-5 may be a promising candidate for advancing to non-human primate and human clinical studies for the development of HIV vaccines based on rBCG vectors. PMID:25255287

  3. In vivo activity of released cell wall lipids of Mycobacterium bovis bacillus Calmette-Guérin is due principally to trehalose mycolates.

    Science.gov (United States)

    Geisel, Rachel E; Sakamoto, Kaori; Russell, David G; Rhoades, Elizabeth R

    2005-04-15

    The hallmark of Mycobacterium-induced pathology is granulomatous inflammation at the site of infection. Mycobacterial lipids are potent immunomodulators that contribute to the granulomatous response and are released in appreciable quantities by intracellular bacilli. Previously we investigated the granulomagenic nature of the peripheral cell wall lipids of Mycobacterium bovis bacillus Calmette-Guérin (BCG) by coating the lipids onto 90-microm diameter microspheres that were mixed into Matrigel matrix with syngeneic bone marrow-derived macrophages and injected i.p. into mice. These studies demonstrated that BCG lipids elicit proinflammatory cytokines and recruit leukocytes. In the current study we determined the lipids responsible for this proinflammatory effect. BCG-derived cell wall lipids were fractionated and purified by liquid chromatography and preparative TLC. The isolated fractions including phosphatidylinositol dimannosides, cardiolipin, phosphatidylglycerol, phosphatidylethanolamine, trehalose monomycolate, trehalose dimycolate, and mycoside B. Trehalose dimycolate, when delivered to bone marrow-derived murine macrophages, induced the greatest secretion of IL-1beta, IL-6, and TNF-alpha in vitro. Trehalose dimycolate similarly induced the greatest secretion of these proinflammatory cytokines in ex vivo matrices over the course of 12 days. Trehalose monomycolate and dimycolate also induced profound neutrophil recruitment in vivo. Experiments with TLR2 or TLR4 gene-deficient mice revealed no defects in responses to trehalose mycolates, although MyD88-deficient mice manifested significantly reduced cell recruitment and cytokine production. These results demonstrate that the trehalose mycolates, particularly trehalose dimycolate, are the most bioactive lipids in the BCG extract, inducing a proinflammatory cascade that influences granuloma formation.

  4. Genotyping and Resolution of a Case of Osteomyelitis in a 16-Month-Old Boy of Hispanic/African American Ethnicity.

    Science.gov (United States)

    Wachira, Eunice; Tran, Kayla; Taylor, Sara; Hoger, Sally; Dunn, James

    2016-02-01

    Most cases of osteomyelitis in children are caused by Staphylococcus aureus, although Kingella kingae, various streptococci, and Salmonella species also underlie this condition. Organisms such as Mycobacterium, Histoplasma, and Cryptococcus are much less commonly identified as etiologic agents in osteomyelitis. This case report describes a 16-month-old boy of Hispanic/African American ethnicity who had extensive inflammation of and discharge from his right ankle. Imaging studies supported a diagnosis of osteomyelitis. Acid-fast bacillus (AFB) and routine wound cultures were ordered on the wound discharge. The AFB culture yielded a positive result for Mycobacterium bovis, and molecular diagnostic testing further genotyped the microorganism as Mycobacterium bovis, Bacillus Calmette-Guerin (BCG). Herein, we report a rare case of osteomyelitis that we believe resulted from a BCG vaccine that the patient had received outside the United States.

  5. Specific T-cell epitopes for immunoassay-based diagnosis of Mycobacterium tuberculosis infection

    DEFF Research Database (Denmark)

    Brock, I; Weldingh, K; Leyten, EM;

    2004-01-01

    Specific T-cell epitopes for immunoassay-based diagnosis of Mycobacterium tuberculosis infection.Brock I, Weldingh K, Leyten EM, Arend SM, Ravn P, Andersen P. Department of Infectious Disease Immunology, Statens Serum Institute, Artillerivej 5, DK-2300 Copenhagen S, Denmark. The currently used...... method for immunological detection of tuberculosis infection, the tuberculin skin test, has low specificity. Antigens specific for Mycobacterium tuberculosis to replace purified protein derivative are therefore urgently needed. We have performed a rigorous assessment of the diagnostic potential of four...... recently identified antigens (Rv2653, Rv2654, Rv3873, and Rv3878) from genomic regions that are lacking from the Mycobacterium bovis bacillus Calmette-Guerin (BCG) vaccine strains as well as from the most common nontuberculous mycobacteria. The fine specificity of potential epitopes in these molecules...

  6. Adverse effects of BCG vaccine 1173 P2 in Iran: A meta-analysis

    Directory of Open Access Journals (Sweden)

    Saied Mostaan

    2016-01-01

    Full Text Available Although in the last two decades the World Health Organization (WHO has introduced tuberculosis as “a threat to global”, the vaccination with the Mycobacterium bovis Bacillus Calmette-Guerin (BCG is the only way for the prevention of this fatal infectious disease. Despite of the efficacy of BCG vaccine especially against infants' meningitis, it has still some limitations due to a variety of adverse effects. Many studies have evaluated the side effects of different strains of BCG vaccines in different countries. In Iran, some studies have been done so far to evaluate the adverse effects of 1173 P2 strain which is used for BCG vaccination. Each of these studies have used different standardization and sampling methods. This review will survey all studies that have been published about adverse effects of 1173 P2 strain of BCG vaccine in Iran using data mining methods.

  7. Antimycobacterial evaluation of novel [4,5-dihydro-1H-pyrazole-1-carbonyl]pyridine derivatives synthesized by microwave-mediated Michael addition.

    Science.gov (United States)

    Sedighi, Vida; Azerang, Parisa; Sardari, Soroush

    2015-06-01

    The focus of this study is the synthesis and biological activity evaluation of a series of dibenzalaceton derivatives (3a-3n) and novel [4,5-dihydro-1H-pyrazole-1-carbonyl]pyridine derivatives (5a-5g) against Mycobacterium bovis, Bacillus Calmette-Guerin (BCG). Dibenzalacetone derivatives were synthesized by benzaldehyde derivatives. The [4,5-dihydro-1H-pyrazole-1-carbonyl]pyridine derivatives were synthesized by Michael addition reaction and using green chemistry microwave-mediated method. All compounds were evaluated against BCG and the activity expressed as minimum inhibitory concentration (MIC) in μM. The result showed good activity for all the compounds especially compounds (3a), (3n), and (5a) illustrated high activity (7.03, 8.10 and 5.37 μM, respectively).

  8. Adverse effects of BCG vaccine 1173 P2 in Iran: A meta-analysis

    Science.gov (United States)

    Mostaan, Saied; Yazdanpanah, Bahador; Moukhah, Rasool; Hozouri, Hamid Reza; Rostami, Manouchehr; Khorashadizadeh, Mohsen; Zerehsaz, Javad; Mahabadi, Ramin Pirhajati; Saadi, Arya; Khanahmad, Hossein; Pooya, Mohammad

    2016-01-01

    Although in the last two decades the World Health Organization (WHO) has introduced tuberculosis as “a threat to global”, the vaccination with the Mycobacterium bovis Bacillus Calmette-Guerin (BCG) is the only way for the prevention of this fatal infectious disease. Despite of the efficacy of BCG vaccine especially against infants’ meningitis, it has still some limitations due to a variety of adverse effects. Many studies have evaluated the side effects of different strains of BCG vaccines in different countries. In Iran, some studies have been done so far to evaluate the adverse effects of 1173 P2 strain which is used for BCG vaccination. Each of these studies have used different standardization and sampling methods. This review will survey all studies that have been published about adverse effects of 1173 P2 strain of BCG vaccine in Iran using data mining methods. PMID:27376038

  9. Combining blue native polyacrylamide gel electrophoresis with liquid chromatography tandem mass spectrometry as an effective strategy for analyzing potential membrane protein complexes of Mycobacterium bovis bacillus Calmette-Guérin

    Directory of Open Access Journals (Sweden)

    Li Weijun

    2011-01-01

    Full Text Available Abstract Background Tuberculosis is an infectious bacterial disease in humans caused primarily by Mycobacterium tuberculosis, and infects one-third of the world's total population. Mycobacterium bovis bacillus Calmette-Guérin (BCG vaccine has been widely used to prevent tuberculosis worldwide since 1921. Membrane proteins play important roles in various cellular processes, and the protein-protein interactions involved in these processes may provide further information about molecular organization and cellular pathways. However, membrane proteins are notoriously under-represented by traditional two-dimensional polyacrylamide gel electrophoresis (2-D PAGE and little is known about mycobacterial membrane and membrane-associated protein complexes. Here we investigated M. bovis BCG by an alternative proteomic strategy coupling blue native PAGE to liquid chromatography tandem mass spectrometry (LC-MS/MS to characterize potential protein-protein interactions in membrane fractions. Results Using this approach, we analyzed native molecular composition of protein complexes in BCG membrane fractions. As a result, 40 proteins (including 12 integral membrane proteins, which were organized in 9 different gel bands, were unambiguous identified. The proteins identified have been experimentally confirmed using 2-D SDS PAGE. We identified MmpL8 and four neighboring proteins that were involved in lipid transport complexes, and all subunits of ATP synthase complex in their monomeric states. Two phenolpthiocerol synthases and three arabinosyltransferases belonging to individual operons were obtained in different gel bands. Furthermore, two giant multifunctional enzymes, Pks7 and Pks8, and four mycobacterial Hsp family members were determined. Additionally, seven ribosomal proteins involved in polyribosome complex and two subunits of the succinate dehydrogenase complex were also found. Notablely, some proteins with high hydrophobicity or multiple transmembrane

  10. Expression and immunogenicity of recombinant Mycobacterium bovis Bacillus Calmette-Guérin strains secreting the antigen ESAT-6 from Mycobacterium tuberculosis in mice

    Institute of Scientific and Technical Information of China (English)

    WANG Li-mei; SHI Chang-hong; FAN Xiong-lin; XUE Ying; BAI Yin-lai; XU Zhi-kai

    2007-01-01

    Background Tuberculosis remains the leading cause of human death. Currently, Bacillus Calmette-Guérin (BCG) is the only available vaccine against tuberculosis but its efficacy is highly variable. Thus, developing new tuberculosis vaccines becomes an urgent task. In this study, we evaluated in BALB/c mice the humoral and cellular immune responses of recombinant BCG expressing the antigen ESAT-6 from Mycobacterium tuberculosis.Methods Escherichia coli-BCG shuttle plasmid named pDE22-esat-6 was constructed by inserting the BamHI/EcoRI digested esat-6 gene PCR product into the similarly digested parental plasmid pDE22. BCG cells were transformed with pDE22-esat-6, which was named recombinant BCG (rBCG). BALB/c mice were immunized subcutaneously on the back with 100 μl normal saline containing 106 CFU of BCG or rBCG. They were sacrificed after 4 weeks to detect their humoral and cellular responses.Results There was no any significant differences in the growth characteristics between the conventional BCG and rBCG. In immunized mice, the IgG antibody titres of rBCG group were as high as 1:8000, which was significantly higher than that in BCG group (1:1400, P<0.05). The elicited IFN-γ level of rBCG group was (1993±106) pg/ml, which was also significantly higher than that in BCG group ((1463±105) pg/ml, P<0.05). The splenocyte proliferation index of rBCG group reached 4.34±0.31, which was higher than that of BCG group (3.79±0.24, P<0.05).Conclusion rBCG secreted expressing antigen ESAT-6 stimulated stronger humoral and cellular immune responses than BCG did, and, therefore may be the better vaccine against mycobacterium tuberculosis.

  11. Autophagy Controls BCG-Induced Trained Immunity and the Response to Intravesical BCG Therapy for Bladder Cancer

    NARCIS (Netherlands)

    Buffen, Kathrin; Oosting, Marije; Quintin, Jessica; Ng, Aylwin; Kleinnijenhuis, Johanneke; Magadi Gopalaiah, Vinod Kumar; van de Vosse, Esther; Wijmenga, Cisca; van Crevel, Reinout; Oosterwijk, Egbert; Grotenhuis, Anne J.; Vermeulen, Sita H.; Kiemeney, Lambertus A.; van de Veerdonk, Frank L.; Chamilos, Georgios; Xavier, Ramnik J.; van der Meer, Jos W. M.; Netea, Mihai G.; Joosten, Leo A. B.

    2014-01-01

    The anti-tuberculosis-vaccine Bacillus Calmette-Guerin (BCG) is the most widely used vaccine in the world. In addition to its effects against tuberculosis, BCG vaccination also induces non-specific beneficial effects against certain forms of malignancy and against infections with unrelated pathogens

  12. Immunisation of colorectal cancer patients with autologous tumour cells

    DEFF Research Database (Denmark)

    Diederichsen, Axel Cosmus Pyndt; Stenholm, A C; Kronborg, O

    1998-01-01

    Patients with colorectal cancer were entered into a clinical phase I trial of immunotherapy with an autologous tumour cell/bacillus Calmette-Guerin (BCG) vaccine. We attempted to describe the possible effects and side effects of the immunisation, and further to investigate whether expression...

  13. Effect of high-dose vitamin A supplementation on the immune response to Bacille Calmette-Guerin vaccine

    DEFF Research Database (Denmark)

    Diness, Birgitte R; Fisker, Ane B; Roth, Adam;

    2007-01-01

    objective was to examine whether VAS influences the immune response to simultaneously administered BCG vaccine. DESIGN: Within a randomized trial of 50,000 IU vitamin A or placebo given with BCG vaccine at birth in Guinea-Bissau, 2710 infants were examined for BCG scar formation and delayed-type...... scar was not affected by VAS. The ex vivo interferon-gamma response to PPD was increased by VAS (means ratio: 1.40; 95% CI: 1.03, 1.91). CONCLUSIONS: VAS with BCG vaccination does not appear to interfere with the long-term immune response to BCG. However, VAS temporarily altered the DTH reaction to PPD...

  14. Defined medium for Moraxella bovis.

    Science.gov (United States)

    Juni, E; Heym, G A

    1986-10-01

    A defined medium (medium MB) for Moraxella bovis was formulated. Nineteen strains grew well on medium MB. One strain was auxotrophic for asparagine, and another was auxotrophic for methionine. Strains of M. equi and M. lacunata also grew on medium MB. All strains had an absolute requirement for thiamine and were stimulated by or actually required the other growth factors in the medium.

  15. Defined medium for Moraxella bovis.

    OpenAIRE

    1986-01-01

    A defined medium (medium MB) for Moraxella bovis was formulated. Nineteen strains grew well on medium MB. One strain was auxotrophic for asparagine, and another was auxotrophic for methionine. Strains of M. equi and M. lacunata also grew on medium MB. All strains had an absolute requirement for thiamine and were stimulated by or actually required the other growth factors in the medium.

  16. Comparison of antigen-specific T-cell responses of tuberculosis patients using complex or single antigens of Mycobacterium tuberculosis

    DEFF Research Database (Denmark)

    Mustafa, A S; Amoudy, H A; Wiker, H G;

    1998-01-01

    GroES, rPstS, rGroEL and rDnaK) antigens of Mycobacterium tuberculosis. The responses of PBMC to these defined antigens were compared with the corresponding results obtained with complex antigens, such as whole-cell M. tuberculosis, M. tuberculosis culture filtrate (MT-CF) and cell wall antigens, as well...... as the vaccine strain, Mycobacterium bovis bacillus Calmette-Guerin (BCG). In addition, M. tuberculosis and MT-CF-induced T-cell lines were tested in the same assays against the panel of purified and complex antigens. The compiled data from PBMC and T-cell lines tested for antigen-induced proliferation and IFN......We have screened peripheral blood mononuclear cells (PBMC) from tuberculosis (TB) patients for proliferative reactivity and interferon-gamma (IFN-gamma) secretion against a panel of purified recombinant (r) and natural (n) culture filtrate (rESAT-6, nMPT59, nMPT64 and nMPB70) and somatic-derived (r...

  17. Síntese e avaliação farmacológica de N’-benzilideno-cumarinas-3-carboidrazidas frente ao Mycobacterium causador da tuberculose

    Directory of Open Access Journals (Sweden)

    Felipe R. Vicente

    2010-04-01

    Full Text Available This article describes a series of twenty-one N’-benzylidene-2-oxo-2H-chromene-3-carbohydrazides (5a-5v which were synthesized and evaluated for their in vitro antibacterial activity against Mycobacterium tuberculosis H37Rv using the Alamar Blue susceptibility test (MABA and the activity expressed as the minimum inhibitory concentration (MIC in μg/mL. Afterwards, the compounds which showed antitubercular activity (50-100 μg/mL 5c, 5g, 5h, 5j, 5l, 5p and 5t were evaluated for their cell viabilities in macrophages infected and non infected with Mycobacterium bovis Bacillus Calmette-Guerin (BCG. The compounds 5c, 5g, 5h, 5j, 5l were not cytotoxic in their respective MIC values, indicating that these compounds could be considered a good starting point to further studies aiming to develop a new lead compound to treat multidrug-resistant tuberculosis (TB-MDR/XDR.

  18. Characterization of the fibronectin-attachment protein of Mycobacterium avium reveals a fibronectin-binding motif conserved among mycobacteria.

    Science.gov (United States)

    Schorey, J S; Holsti, M A; Ratliff, T L; Allen, P M; Brown, E J

    1996-07-01

    Mycobacterium avium is an intracellular pathogen and a major opportunistic infectious agent observed in patients with acquired immune deficiency syndrome (AIDS). Evidence suggests that the initial portal of infection by M. avium is often the gastrointestinal tract. However, the mechanism by which the M. avium crosses the epithelial barrier is unclear. A possible mechanism is suggested by the ability of M. avium to bind fibronectin, an extracellular matrix protein that is a virulence factor for several extracellular pathogenic bacteria which bind to mucosal surfaces. To further characterize fibronectin binding by M. avium, we have cloned the M. avium fibronectin-attachment protein (FAP). The M. avium FAP (FAP-A) has an unusually large number of Pro and Ala residues (40% overall) and is 50% identical to FAP of both Mycobacterium leprae and Mycobacterium tuberculosis. Using recombinant FAP-A and FAP-A peptides, we show that two non-continuous regions in FAP-A bind fibronectin. Peptides from these regions and homologous sequences from M. leprae FAP inhibit fibronectin binding by both M. avium and Mycobacterium bovis Bacillus Calmette-Guerin (BCG). These regions have no homology to eukaryotic fibronectin-binding proteins and are only distantly related to fibronectin-binding peptides of Gram-positive bacteria. Nevertheless, these fibronectin-binding regions are highly conserved among the mycobacterial FAPs, suggesting an essential function for this interaction in mycobacteria infection of their metazoan hosts.

  19. Comparison of antigen-specific T-cell responses of tuberculosis patients using complex or single antigens of Mycobacterium tuberculosis

    DEFF Research Database (Denmark)

    Mustafa, A S; Amoudy, H A; Wiker, H G

    1998-01-01

    We have screened peripheral blood mononuclear cells (PBMC) from tuberculosis (TB) patients for proliferative reactivity and interferon-gamma (IFN-gamma) secretion against a panel of purified recombinant (r) and natural (n) culture filtrate (rESAT-6, nMPT59, nMPT64 and nMPB70) and somatic-derived (r......GroES, rPstS, rGroEL and rDnaK) antigens of Mycobacterium tuberculosis. The responses of PBMC to these defined antigens were compared with the corresponding results obtained with complex antigens, such as whole-cell M. tuberculosis, M. tuberculosis culture filtrate (MT-CF) and cell wall antigens, as well...... as the vaccine strain, Mycobacterium bovis bacillus Calmette-Guerin (BCG). In addition, M. tuberculosis and MT-CF-induced T-cell lines were tested in the same assays against the panel of purified and complex antigens. The compiled data from PBMC and T-cell lines tested for antigen-induced proliferation and IFN...

  20. Disseminated BCG infection in a patient with severe combined immunodeficiency

    Energy Technology Data Exchange (ETDEWEB)

    Han, Tae Il [Eulji University School of Medicine, Taejon (Korea, Republic of); Kim, In One; Kim, Woo Sun; Yeon, Kyung Mo [Seoul National University College of Medicine, Seoul (Korea, Republic of)

    2000-06-01

    Disseminated mycobacterial infection after bacillus Calmette-Guerin (BCG) accination is a very rare disorder, occurring mostly in patients with immunologic eficiency. We report a case of disseminated BCG infection in a 16-month-old girl with severe combined immunodeficiency. Plain radiographs showed multiple osteolytic lesions in the femora, tibiae, humerus, and phalanges. Abdominal sonography and CT scanning revealed multiple nodules in the spleen, and portocaval lymphadenopathy.

  1. Disseminated BCG Infection in a patient with Severe Combined Immunodeficiency

    OpenAIRE

    Han, Tae Il; Kim, In-One; Kim, Woo Sun; Yeon, Kyung Mo

    2000-01-01

    Disseminated mycobacterial infection after bacillus Calmette-Guerin (BCG) vaccination is a very rare disorder, occurring mostly in patients with immunologic deficiency. We report a case of disseminated BCG infection in a 16-month-old girl with severe combined immunodeficiency. Plain radiographs showed multiple osteolytic lesions in the femora, tibiae, humerus, and phalanges. Abdominal sonography and CT scanning revealed multiple nodules in the spleen, and portocaval lymphadenopathy.

  2. THE CONSTRUCTION AND EXPRESSION OF RECOMBINANT SHUTTLE PLASMID WITH OMPL1 GENE FROM LEPTOSPIRA INTERROGANS SEROVAR LAI STRAIN 017 IN BACILLE CALMETTE GUERIN

    Institute of Scientific and Technical Information of China (English)

    鲍朗; 邱洪宇; 晏菊芳; 谢勇恩; 陈玮

    2002-01-01

    Objective.To construct recombinant BCG against leptospirosis.Methods.We amplified the entire open reading frame of the OmpL1 gene from the genome of the leptospire serovar Lai strain 017.Two recombinant plasmids pBQ1 and pBQ2 were constructed by oriented ligation based on the E.coli BCG shuttle plasmids pMV261 and pMV361 respectively.The recombinant plasmids were transformed into BCG by electroporation.The rBCGs bearing pBQ1 and pBQ2 were induced by high temperature of 45℃ .Results.The expressed product,a 35kD protein was detected by SDS PAGE.The result indicates that pBQ1 and pBQ2 can express OmpL1 in rBCG.Conclusion.The technical methods in this study may help detect the immunogenicity and immunoprotection of OmpL1 and develop more safe,highly effective rBCG bearing leptospiral antigen with long lasting protection.

  3. Accuracy of the QuantiFERON-TB Gold in Tube for diagnosing tuberculosis in a young pediatric population previously vaccinated with Bacille Calmette-Guerin

    Directory of Open Access Journals (Sweden)

    Marcelo Genofre Vallada

    2014-03-01

    Full Text Available Objective: To evaluate the accuracy of an interferongamma release assay (QuantiFERON-TB Gold in Tube for diagnosing Mycobacterium tuberculosis infection in a young pediatric population. Methods: 195 children previously vaccinated with BCG were evaluated, being 184 healthy individuals with no clinical or epidemiological evidence of mycobacterial infection, and 11 with Mycobacterium tuberculosis infection, according to clinical, radiological, and laboratory parameters. A blood sample was obtained from each child and processed according to the manufacturer's instructions. The assay performance was evaluated by a Receiver Operating Characteristic (ROC curve. Results: In the group of 184 non-infected children, 130 (70.6% were under the age of four years (mean age of 35 months. In this group, 177 children (96.2% had negative test results, six (3.2% had indeterminate results, and one (0.5% had a positive result. In the group of 11 infected children, the mean age was 58.5 months, and two of them (18% had negative results. The ROC curve had an area under the curve of 0.88 (95%CI 0.82-0.92; p<0.001, disclosing a predictive positive value of 81.8% for the test (95%CI 46.3-97.4. The assay sensitivity was 81.8% (95%CI 48.2-97.2 and the specificity was 98.8% (95%CI 96-99.8. Conclusions: In the present study, the QuantiFERON-TB Gold in Tube performance for diagnosing M. tuberculosis infection was appropriate in a young pediatric population.

  4. Perbedaan Kadar Interferon Gamma dan Interleukin-10 pada Orang Dewasa Terinfeksi Ascaris Lumbricoides dengan Tidak Terinfeksi yang Diinduksi Vaksin Bacille Calmette-Guerin

    OpenAIRE

    Weni Mulyani; Nuzulia Irawati; Netti Suharti

    2016-01-01

    AbstrakKecacingan merupakan penyakit yang masih banyak di negara berkembang. Penyakit ini dapat menimbulkan gangguan gizi, pertumbuhan dan penurunan produktifitas kerja. Infeksi cacing dapat menimbulkan penurunan respon terhadap antigen sebagai akibat modified Th2 response. Vaksin BCG merupakan antigen yang dikenal sebagai penginduksi respon sel Th1. Tujuan penelitian ini adalah menentukan perbedaan kadar IFN-γ dan IL-10 antara orang dewasa terinfeksi Ascaris lumbricoides dan tidak terinfeksi...

  5. Molecular epidemiology of human cases of tuberculosis by Mycobacterium bovis in Mexico.

    Science.gov (United States)

    Milián-Suazo, Feliciano; Pérez-Guerrero, Laura; Arriaga-Díaz, Camila; Escartín-Chávez, Minerva

    2010-10-01

    The purpose of this study was to evaluate the role of Mycobacterium bovis in human cases of tuberculosis (TB) in an endemic area of the disease in cattle. Sputum, urine and other tissue samples were obtained from: (1) TB-symptomatic patients, (2) dairy farm workers and (3) abattoir workers. Samples of macroscopic lesions suspicious of TB were also obtained from cattle at slaughter in the same geographic area. A total of 562 human samples were collected: 255 from symptomatic patients, 218 from farm workers and 93 from abattoir workers. Samples were analysed by the bacillus acido-alcohol resitant (BAAR) and polymerase chain reaction (PCR) tests and cultured in Stonebrink and Löwenstein-Jensen. Spoligotyping was performed in all isolates obtained by culture and the DNA obtained by PCR. From the total number of human cases, 34 (6%) showed M. bovis spoligotype; eight spoligotypes from cattle showed an identical pattern to three spoligotypes from humans; a different set of spoligotypes from cattle (n = 8) had only one spacer difference to a set of spoligotypes from humans (n = 2). These results provide further evidence that infected cattle represent a risk to public health and support previous reports about the role of M. bovis in Mexican patients. There is no doubt that genotyping M. bovis isolates collected from cattle may have a substantial impact on our understanding of the epidemiology of TB.

  6. Mycobacterium bovis (Bovine Tuberculosis) in Humans

    Science.gov (United States)

    ... disease is treated with a combination of several antibiotics. Latent infection without disease is not treated with pyrazinamide. Are all cattle infected with M. bovis? No. The Cooperative State- ...

  7. A Literary Study on Bezoar Bovis

    Directory of Open Access Journals (Sweden)

    Seong Kyu Park

    2005-12-01

    Full Text Available Objectives : Literary investigation of existing data was conducted to verify effects of Bezoar Bovis and its herbal acupuncture, and determine quality management through component analysis. Results : Following results were obtained through literary investigation. 1. Bezoar Bovis is dried cystic stone from a cattle. Its characteristics are cool, no toxicity, and bitter taste. Known actions are: quells heat and detoxifies Fire Poison, extinguishes internal movement of Liver Wind and stops convulsion, vaporizes phlegm, and opens orifice. It is mainly used for treating tremor, stroke, delirium, sore throat, oral furuncle, boil, and others. 2. Bezoar Bovis is effective for eliminating liver toxicity, protecting against brain damage, and has anti-microbial activities. 3. Bezoar Bovis is mixed with bear gall bladder and deer musk to be used as herbal acupuncture, and this mixture is effective is invigorating liver functions as well as treating arthritis, headache, and etc. 4. Principal components of Bezoar Bovis are bilirubin-type pigments and cholic acids. The amount of bilirubin can be used as a standard to determine the quality of Bezoar Bovis.

  8. Isolamento de Mycobacterium bovis em cão Mycobacterium bovis isolation in a dog

    Directory of Open Access Journals (Sweden)

    P.M.P.C. Mota

    2001-08-01

    Full Text Available This report describes the isolation of Mycobacterium bovis from a dog with a history of co-habitation with bufallos infected with Mycobacterium bovis. After necropsy, the microrganism was isolated from a mesenteric lymphatic node in Stonebrink media and bacterial identification was confirmed by biochemical tests.

  9. Abortion associated with Mycoplasma bovis (M. bovis) in a bison (Bison bison) herd

    Science.gov (United States)

    Mycoplasma bovis (M. bovis) has recently emerged as a significant health threat in bison and is an increasing concern and source of economic loss for producers. Clinical manifestations of infection documented in bison include pneumonia, respiratory distress and polyarthritis. The current study des...

  10. Nanoparticulation of BCG-CWS for application to bladder cancer therapy

    OpenAIRE

    Nakamura, Takashi; Fukiage, Masafumi; Higuchi, Megumi; NAKAYA, Akihiro; Yano, Ikuya; Miyazaki, Jun; Nishiyama, Hiroyuki; Akaza, Hideyuki; Ito, Toshihiro; Hosokawa, Hiroyuki; Nakayama, Toshinori; Harashima, Hideyoshi

    2014-01-01

    The Mycobacterium bovis Bacille Calmette-Guerin cell wall skeleton (BCG-CWS) could be used to replace live BCG as a bladder cancer drug. However, because BCG-CWS is poorly soluble, has a strong-negative charge, very high molecular weight and heterogeneity in size of tens of mu m, it cannot be used in such an application. We report herein on the development of a novel packaging method that permits BCG-CWS to be encapsulated into 166 nm-sized lipid particles. The BCG-CWS encapsulated nano parti...

  11. Lack of transplacental transmission of Bartonella bovis.

    Science.gov (United States)

    Chastant-Maillard, S; Boulouis, H-J; Reynaud, K; Thoumire, S; Gandoin, C; Bouillin, C; Cordonnier, N; Maillard, R

    2015-02-01

    Transplacental transmission of Bartonella spp. has been reported for rodents, but not for cats and has never been investigated in cattle. The objective of this study was to assess vertical transmission of Bartonella in cattle. Fifty-six cow-calf pairs were tested before (cows) and after (calves) caesarean section for Bartonella bacteremia and/or serology, and the cotyledons were checked for gross lesions and presence of the bacteria. None of the 29 (52%) bacteremic cows gave birth to bacteremic calves, and all calves were seronegative at birth. Neither placentitis nor vasculitis were observed in all collected cotyledons. Bartonella bovis was not detected in placental cotyledons. Therefore, transplacental transmission of B. bovis and multiplication of the bacteria in the placenta do not seem likely. The lack of transplacental transmission may be associated with the particular structure of the placenta in ruminants or to a poor affinity/agressiveness of B. bovis for this tissue.

  12. Plasmid profiles of Moraxella bovis isolates.

    Science.gov (United States)

    McDonald, T J; Pugh, G W

    1986-04-01

    Two-hundred isolates of Moraxella bovis were selected at random and examined for the presence of plasmid DNA by a rapid alkaline-detergent lysis method. All isolates contained from 1 to 6 plasmids, with varying agarose-gel electrophoretic migration patterns. Most (80%) isolates carried 2 to 4 plasmids, which ranged in molecular weight from 2.6 to 80 megadaltons. Seemingly, plasmid profiles can be used as a simple, reliable epizootiologic tool to establish a strain identification scheme for M bovis.

  13. Bacillus coagulans

    Science.gov (United States)

    Bacillus coagulans is a type of bacteria. It is used similarly to lactobacillus and other probiotics as "beneficial" bacteria. People take Bacillus coagulans for diarrhea, including infectious types such as ...

  14. Bacteremia with the bovis group streptococci

    DEFF Research Database (Denmark)

    Marmolin, Ea S; Hartmeyer, Gitte N; Christensen, Jens J;

    2016-01-01

    DNA sequencing of the intergenic spacer (ITS) region was used to identify 53 blood culture isolates that had previously been designated to the bovis group streptococci and clinical data was collected retrospectively from patients' records using a standardized protocol. ITS sequencing identified 1...

  15. Invasion of erythrocytes by Babesia bovis

    NARCIS (Netherlands)

    Gaffar, Fasila Razzia

    2004-01-01

    In this thesis we investigated the invasion of erythrocytes taking place during the asexual erythrocytic blood stage of the apicomplexan parasites Babesia bovis parasite. Host cell invasion by apicomplexan parasites is a complex process requiring multiple receptor-ligand interactions, involving ass

  16. Partial characterization of a Moraxella bovis cytolysin.

    Science.gov (United States)

    Gray, J T; Fedorka-Cray, P J; Rogers, D G

    1995-02-01

    Moraxella bovis (M. bovis) is the etiologic agent of infectious bovine keratoconjunctivitis and M. bovis hemolysin is believed to be an important virulence factor. Two strains of M. bovis were compared, Epp 63(300) (Epp), a known virulent and hemolytic strain, and IBH 63 (IBH), a known avirulent and nonhemolytic strain. Sterile 10-fold (10x) supernatant concentrates were obtained from cultures grown in TSB broth with 10 mM CaCl2. Supernatant hemolysin titers for Epp, were 1:1024 and 1:8192 for unconcentrated (1x) and 10x, respectively. Supernatant cytotoxin titers to bovine mononuclear cells were 1:32 and 1:128 for 1x and 10x, respectively, for Epp. Cytolytic (hemolytic and cytotoxic) activities declined 10-fold but were still measurable for > 1 wk at 4 degrees C. Both activities were inactivated by trypsin and by heating at 56 degrees C for 20 min. A cytotoxic effect was observed on cultured bovine and ovine corneal epithelial cells with Epp. All cytolytic effects were neutralized with antiserum to 10x Epp. No cytolytic activities were detected for 10x IBH. SDS-PAGE electrophoresis and related immunoblots indicate a high molecular weight protein at 110 kDa for the 10x Epp preparation when stained with silver or probed with monoclonal antibodies to the E. coli alpha hemolysin. No 110 kDa band is observed for 10x IBH. These data suggest that hemolytic and cytotoxic activities are important in the pathogenesis of infectious bovine keratoconjunctivitis and identify the protein as a possible RTX related toxin of 110 kDa. Stability of the M. bovis cytolysin for > 1 week should allow further characterization and purification of the protein.

  17. Effectiveness of transurethral resection under the control of photodynamic diagnosis and intravesical instillation of bacillus Calmette–Guérin in case of poorly differentiated non-muscle-invasive bladder cancer

    Directory of Open Access Journals (Sweden)

    A. I. Rolevich

    2016-01-01

    Full Text Available Background. High-grade non-muscle-invasive bladder cancer (NMIBC is characterized by a high rate of recurrence, progression, and mortality associated with this disease. Organ-preserving treatment by transurethral resection and immunotherapy with bacillus Calmette-Guerin (BCG is an initial approach to therapy in these patients. However, the efficacy of such therapy is limited. This justifies the use of other methods of treatment, such as TUR under the control of photodynamic diagnosis (PDD. Aim of this study was to evaluate the effectiveness of therapeutic interventions in patients with high-grade NMIBC.Materials and methods. We have retrospectively analyzed results of follow-up of patients with primary or recurrent high-grade transitional cell NMIBC, treatment by TUR in conjunction with BCG or without it N.N. Alexandrov National Cancer Centre in the period from 2004 to 2013. In total, the study included 113 patients (27 women and 86 men, in the median age of 72 years. We have evaluated 5-year recurrence- and progression-free survival, analyzed an influence of prognostic factors and methods of treatment on the risk of recurrence and progression with Cox model and Kaplan–Meier method.Results. With a median of follow up of 59 (12–116 months the rates of 5-year recurrence- and progression-free survival were respectively 42.5 and 71.6 %. Statistically significant association with the risk of recurrence was observed in multivariate Cox regression analysis for recurrent tumors (hazard ratio (HR 2.73; 95 % confidence interval (CI 1.61–4.62 and immunotherapy with BCG (HR 0.56; 95 % CI 0.31–0.99. BCG significantly increased recurrence-free survival in patients with both primary tumors, and with recurrent ones. Significant factors in the multivariate analysis with regard to the risk of progression were suspicion for muscle-invasive tumors according to the cystoscopic picture (HR 3.36; 95 % CI 1.09–10.4, abnormal tumor-free bladder mucosa

  18. Identification of a capsular polysaccharide from Moraxella bovis.

    Science.gov (United States)

    Wilson, Jennifer C; Hitchen, Paul G; Frank, Martin; Peak, Ian R; Collins, Patrick M; Morris, Howard R; Dell, Anne; Grice, I Darren

    2005-03-21

    The bacterium Moraxella bovis is the causative agent of an economically important disease of cattle: Infectious Bovine Keratoconjunctivitis (IBK), otherwise known as pinkeye. Little is known regarding the structure of the carbohydrates produced by M. bovis. The structure of a capsular polysaccharide from M. bovis (strain Mb25) has been determined using NMR and MS analysis. From these data it is concluded that the polysaccharide is composed of the unmodified chondroitin disaccharide repeat unit.

  19. A live attenuated BCG vaccine overexpressing multistage antigens Ag85B and HspX provides superior protection against Mycobacterium tuberculosis infection.

    Science.gov (United States)

    Yuan, Xuefeng; Teng, Xindong; Jing, Yukai; Ma, Jilei; Tian, Maopeng; Yu, Qi; Zhou, Lei; Wang, Ruibo; Wang, Weihua; Li, Li; Fan, Xionglin

    2015-12-01

    Tuberculosis (TB) remains one of the most menacing infectious diseases, although attenuated Mycobacterium bovis Bacillus Calmette-Guerin (BCG) vaccine has been widely used to protect children against primary TB. There are increasing evidences that rapid growing and dormant Mycobacterium tuberculosis (M. tuberculosis) coexist in vivo after infection. However, BCG vaccine only elicits cell-mediated immune responses to secretory antigens expressed by rapid growing pathogen. BCG vaccine is thus unable to thwart the reactivation of latent tuberculosis infection (LTBI), and its protection wanes over age after neonatal immunization. In order to extend its ability for a durable protection, a novel recombinant BCG (rBCG) strain, named rBCG::XB, was constructed by overexpressing immunodominant multistage antigens of Ag85B and HspX, which are expressed by both rapid replicating and dormant M. tuberculosis. Long-term protective effect and immunogenicity of rBCG::XB were compared with the parental BCG in vaccinated C57BL/6 mice. Our results demonstrated that rBCG::XB provided the stronger and long-lasting protection against M. tuberculosis H37Rv intranasal infection than BCG. The rBCG::XB not only elicited the more durable multistage antigen-specific CD4(+)Th1-biased immune responses and specific polyfunctional CD4(+)T cells but also augmented the CD8(+) CTL effects against Ag85B in vivo. In particular, higher levels of CD4(+) TEM and CD8(+) TCM cells, dominated by IL2(+) CD4(+) and CD8(+) TCM cells, were obtained in the spleen of rBCG::XB vaccinated mice. Therefore, our findings indicate that rBCG::XB is a promising candidate to improve the efficacy of BCG.

  20. A recombinant adenovirus expressing CFP10, ESAT6, Ag85A and Ag85B of Mycobacterium tuberculosis elicits strong antigen-specific immune responses in mice.

    Science.gov (United States)

    Li, Wu; Deng, Guangcun; Li, Min; Zeng, Jin; Zhao, Liping; Liu, Xiaoming; Wang, Yujiong

    2014-11-01

    Tuberculosis (TB) is caused by an infection of Mycobacterium tuberculosis (Mtb) and remains an enormous and increasing health burden worldwide. To date, Mycobacterium bovis Bacillus Calmette Guerin (BCG) is the only licensed anti-TB vaccine worldwide, which provides an important but limited protection from the Mtb infection. The development of alternative anti-TB vaccines is therefore urgently needed. Here we report, the generation of Ad5-CEAB, a recombinant adenovirus expressing Mtb antigens of CFP10, ESAT6, Ag85A and Ag85B proteins in a form of mixture. In order to evaluate the immunogenicity of Ad5-CEAB, mice were immunized with Ad5-CEAB by intranasal instillation three times with 2-week intervals. The results demonstrated that Ad5-CEAB elicited a strong antigen-specific immune response, particularly of the Th1 immune responses that were characterized by an increased ratio of IgG2a/IgG1 and secretions of Th1 type cytokines, IFN-γ, TNF-α, IL-2 and IL-12. In addition, the Ad5-CEAB also showed an ability to enhance humoral responses with a dramatically augmented antigen-specific serum IgG. Furthermore, an elevated sIgA were also found in the bronchoalveolar lavage fluid of the immunized mice, suggesting the elicitation of mucosal immune responses. These data indicate that Ad5-CEAB can induce a broad range of antigen-specific immune responses in vivo, which provides a promising and novel route for developing anti-TB vaccines and warrants further investigation.

  1. Immunogenic Properties of a BCG Adjuvanted Chitosan Nanoparticle-Based Dengue Vaccine in Human Dendritic Cells.

    Directory of Open Access Journals (Sweden)

    Taweewun Hunsawong

    2015-09-01

    Full Text Available Dengue viruses (DENVs are among the most rapidly and efficiently spreading arboviruses. WHO recently estimated that about half of the world's population is now at risk for DENV infection. There is no specific treatment or vaccine available to treat or prevent DENV infections. Here, we report the development of a novel dengue nanovaccine (DNV composed of UV-inactivated DENV-2 (UVI-DENV and Mycobacterium bovis Bacillus Calmette-Guerin cell wall components (BCG-CWCs loaded into chitosan nanoparticles (CS-NPs. CS-NPs were prepared by an emulsion polymerization method prior to loading of the BCG-CWCs and UVI-DENV components. Using a scanning electron microscope and a zetasizer, DNV was determined to be of spherical shape with a diameter of 372.0 ± 11.2 nm in average and cationic surface properties. The loading efficacies of BCG-CWCs and UVI-DENV into the CS-NPs and BCG-CS-NPs were up to 97.2 and 98.4%, respectively. THP-1 cellular uptake of UVI-DENV present in the DNV was higher than soluble UVI-DENV alone. DNV stimulation of immature dendritic cells (iDCs resulted in a significantly higher expression of DCs maturation markers (CD80, CD86 and HLA-DR and induction of various cytokine and chemokine productions than in UVI-DENV-treated iDCs, suggesting a potential use of BCG- CS-NPs as adjuvant and delivery system for dengue vaccines.

  2. A novel dengue virus serotype-2 nanovaccine induces robust humoral and cell-mediated immunity in mice.

    Science.gov (United States)

    Hunsawong, Taweewun; Sunintaboon, Panya; Warit, Saradee; Thaisomboonsuk, Butsaya; Jarman, Richard G; Yoon, In-Kyu; Ubol, Sukathida; Fernandez, Stefan

    2015-03-30

    Dengue virus (DENV), a member of the Flaviviridae family, can be transmitted to humans through the bite of infected Aedes mosquitoes. The incidence of dengue has increased worldwide over the past few decades. Inadequate vector control, changing global ecology, increased urbanization, and faster global travel are factors enhancing the rapid spread of the virus and its vector. In the absence of specific antiviral treatments, the search for a safe and effective vaccine grows more imperative. Many strategies have been utilized to develop dengue vaccines. Here, we demonstrate the immunogenic properties of a novel dengue nanovaccine (DNV), composed of ultraviolet radiation (UV)-inactivated DENV-2, which has been loaded into the nanoparticles containing chitosan/Mycobacterium bovis Bacillus Calmette-Guerin cell wall components (CS/BCG-NPs). We investigated the immunogenicity of DNV in a Swiss albino mouse model. Inoculation with various concentrations of vaccine (0.3, 1, 3 and 10μg/dose) with three doses, 15-day apart, induced strong anti-dengue IgM and IgG antibodies in the mouse serum along with neutralizing antibody against DENV-2 reference strain (16681), a clinical-isolate strain (00745/10) and the mouse-adapted New Guinea-C (NGC) strain. Cytokine and chemokine secretion in the serum of DNV-immunized mice showed elevated levels of IFN-γ, IL-2, IL-5, IL-12p40, IL-12p70, IL-17, eotaxin and RANTES, all of which have varying immune functions. Furthermore, we observed a DNV dose-dependent increase in the frequencies of IFN-γ-producing CD4(+) and CD8(+) T cells after in vitro stimulation of nucleated cells. Based on these findings, DNV has the potential to become a candidate dengue vaccine.

  3. Mycobacterium tuberculosis PPD-induced immune biomarkers measurable in vitro following BCG vaccination of UK adolescents by multiplex bead array and intracellular cytokine staining

    Directory of Open Access Journals (Sweden)

    Worth Andrew

    2010-07-01

    Full Text Available Abstract Background The vaccine efficacy reported following Mycobacterium bovis Bacillus Calmette Guerin (BCG administration to UK adolescents is 77% and defining the cellular immune response in this group can inform us as to the nature of effective immunity against tuberculosis. The aim of this study was to identify which cytokines and lymphocyte populations characterise the peripheral blood cellular immune response following BCG vaccination. Results Diluted blood from before and after vaccination was stimulated with Mycobacterium tuberculosis purified protein derivative for 6 days, after which soluble biomarkers in supernatants were assayed by multiplex bead array. Ten out of twenty biomarkers measured were significantly increased (p Mycobacterium tuberculosis purified protein derivative stimulation of PBMC samples from the 12 month group revealed that IFNγ expression was detectable in CD4 and CD8 T-cells and natural killer cells. Polyfunctional flow cytometry analysis demonstrated that cells expressing IFNγ alone formed the majority in each subpopulation of cells. Only in CD4 T-cells and NK cells were there a notable proportion of responding cells of a different phenotype and these were single positive, TNFα producers. No significant expression of the cytokines IL-2, IL-17 or IL-10 was seen in any population of cells. Conclusions The broad array of biomarker responses detected by multiplex bead array suggests that BCG vaccination is capable, in this setting, of inducing a complex immune phenotype. Although polyfunctional T-cells have been proposed to play a role in protective immunity, they were not present in vaccinated adolescents who, based on earlier epidemiological studies, should have developed protection against pulmonary tuberculosis. This may be due to the later sampling time point available for testing or on the kinetics of the assays used.

  4. The zoonotic importance of Mycobacterium bovis.

    Science.gov (United States)

    Moda, G; Daborn, C J; Grange, J M; Cosivi, O

    1996-04-01

    The zoonotic importance of Mycobacterium bovis has been the subject of renewed interest in the wake of the increasing incidence of tuberculosis in the human population. This paper considers some of the conditions under which transmission of M. bovis from animals to humans occurs and reviews current information on the global distribution of the disease. The paper highlights the particular threat posed by this zoonotic disease in developing countries and lists the veterinary and human public health measures that need to be adopted if the disease is to be contained. The association of tuberculosis with malnutrition and poverty has long been recognized and the need to address these basic issues are as crucial as specific measures against the disease itself.

  5. Transmissie van Mycobacterium bovis tussen mens en dier

    NARCIS (Netherlands)

    Vries, de G.; Beer, de J.; Bakker, D.; Soolingen, D.

    2015-01-01

    Nederland is officieel vrij van rundertuberculose. Toch komt af en toe nog Mycobacterium bovis-tuberculose voor bij relatief jonge autochtone Nederlanders. Ook zijn er recent nog wel boviene-uitbraken geweest. Dat roept de vraag op of er ook nu nog transmissie is van M.bovis tussen mens en dier. Daa

  6. Sensitivity of Mycobacterium bovis to common beef processing interventions

    Science.gov (United States)

    Objective. Mycobacterium bovis is the causative agent of bovine tuberculosis, a relevant zoonosis that can spread to humans through inhalation or by ingestion. M. bovis multiplies slowly, so infected animals may be sent to slaughter during the early stages of the disease before diagnosis and when ...

  7. Tuberculosis Caused by Mycobacterium bovis in a Capybara (Hydrochoerus hydrochaeris).

    Science.gov (United States)

    Mol, J P S; Carvalho, T F; Fonseca, A A; Sales, E B; Issa, M A; Rezende, L C; Hodon, M A; Tinoco, H P; Malta, M C C; Pessanha, A T; Pierezan, F; Mota, P M P C; Paixão, T A; Santos, R L

    2016-01-01

    Tuberculosis, associated with Mycobacterium bovis, was diagnosed post mortem in an adult female capybara (Hydrochoerus hydrochaeris), kept at the Pampulha Ecological Park, Belo Horizonte, Brazil, in a large metropolitan area. On post-mortem examination, there were numerous firm white nodules scattered throughout all lobes of both lungs. Tissue samples were collected for histological and microbiological examination. Microscopically, the pulmonary nodules were multifocal to coalescing granulomas and intralesional acid-fast bacilli were evident in Ziehl-Neelsen-stained sections of the lung and spleen. Colonies with morphological features of Mycobacterium spp. were isolated from lung samples and conventional polymerase chain reaction (PCR) with genomic DNA from the isolates was positive for M. bovis; sequencing indicated 100% identity with the region of difference 4 (RD4) of M. bovis. In addition, M. bovis DNA was detected in the lung by quantitative PCR. The finding of M. bovis in a capybara indicates a potential public health risk in a zoological collection.

  8. Intratumoral injection of BCG-CWS-pretreated dendritic cells following tumor cryoablation.

    Science.gov (United States)

    Kawamura, Naoshi; Udagawa, Masaru; Fujita, Tomonobu; Sakurai, Toshiharu; Yaguchi, Tomonori; Kawakami, Yutaka

    2014-01-01

    Intratumoral administration of dendritic cells (DC) following cryoablation of tumor is one of the personalized cancer immunotherapies which is able to induce immune responses to multiple endogenous tumor antigens, including shared and unique antigens. Here we describe protocols of cryoablation of tumors, generation of cultured DC, pretreatment of DC with a Toll-like receptor (TLR)-stimulating purified component of Bacillus Calmette-Guerin cell wall fraction (BCG-CWS) and highly immunogenic keyhole limpet hemocyanin (KLH) antigen, and combined use of tumor cryoablation and intratumoral administration of BCG-CWS-pretreated DC in both a murine model and cancer patients.

  9. [Modifications by BCG of the mortality of the irradiated mouse].

    Science.gov (United States)

    Allain, P; Chaleil, D; Maingot, D; Larra, F

    1976-01-01

    Freeze-dried Bacillus Calmette Guerin (B.C.G.) of Institut Pasteur was given by intravenous route to mice at 1,2 and 4 mg/kg before and after gamma irradiation of animals by 1 000 rad. B.C.G. 1 mg/kg injected the day or the day after irradiation has a protective effect (mortality reduced from 77% for controls to 58% and 50% for treated mice). B.C.G. given before irradiation in single or double doses increased mortality.

  10. Meta-analysis of the Risk Factor of Tuberculosis in China%中国人群肺结核发病危险因素的meta分析

    Institute of Scientific and Technical Information of China (English)

    李斌; 王兆芬; 刘寿; 杜文琪

    2011-01-01

    Objective To explore the main risk factors related to the incidence of tuberculosis in China and to provide evidence for prevention. Methods The results of 19 studies on the main risk factors of tuberculosis were analyzed by Meta-analysis method. Results The pooled odds radio values and 95% CI of history of exposure to pulmonary tuberculosis,Bacillus Calmette Guerin vaccination,indoor conditions, work environment,Bacillus Calmette Guerin vaccination scar and body mass index (BMI) were 2.87 (95% CI, 2.37 -3.49), 0.41 (95%CI:0.34 ~ 0.51),2.62(95% CI: 1.96 -3.50) ,3.20(95%CI:2.44 -4.20),2.86 (95%CI:1.94 -4.20) ,2.32(95% CI-1. 15 ~4.70). Conclusion The main risk factors of were the history of exposure to pulmonary tuberculosis, indoor conditions,work environment,Bacillus Calmette Guerin vaccination scar and body mass index(BMI) .while Bacillus Calmette Guerin vaccination was a protect factor.%目的 探讨我国人群肺结核病发病的主要危险因素,为今后防治工作提供依据.方法 利用meta分析方法分析国内19篇关于肺结核病发病危险因素的研究文献.结果结核病人接触史、卡介苗接种史、室内环境、工作环境、卡痕和BMI的合并OR值(95%CI)分别为:2.87(2.37 ~ 3.49),0.41(0.34 ~0.51),2.62(1.96 ~3.50),3.20(2.44~4.20),2.86( 1.94 ~4.20),2.32(1.15 ~4.70).结 论结核病人接触史、室内环境、工作环境、卡痕和BMI是我国肺结核病发病的主要危险因素,卡介苗接种为保护性因素.

  11. BCG plus levamisole following irradiation of advanced squamous bronchial carcinoma. [Hard X Radiation

    Energy Technology Data Exchange (ETDEWEB)

    Pines, A.

    1980-08-01

    Fifty patients with inoperable squamous cell carcinoma of the bronchus were treated with radical radiotherapy. Afterwards, 16 patients received levamisole on 2 days per week and bacillus calmette guerin (B.C.G.) skin innoculations every two weeks;another 16 received the same dosage of levamisole but B.C.G. every 4 weeks; 18 patients were controls. Survival was better in the first group of patients only during the first two years of study (P = 0.02) but not later: metastases were fewer. Both B.C.G. and levamisole gave little discomfort when the dose was adjusted for each patient.

  12. Mastite bovina por Mycoplasma bovis em rebanhos leiteiros Mastitis caused by Mycoplasma bovis in dairy cattle

    Directory of Open Access Journals (Sweden)

    Lucienne G. Pretto

    2001-12-01

    Full Text Available Foram examinadas 713 vacas de três rebanhos leiteiros localizados na região norte do Estado do Paraná e sudoeste do Estado de São Paulo, das quais 137 apresentaram mastite. Nas três propriedades foram detectados oito animais (1,12% com mastite clínica por Mycoplasma bovis. Destes animais, quatro tratados com oxitetraciclina e tilosina e três com enrofloxacina, não responderam ao tratamento e foram descartados no decorrer da lactação. Uma vaca medicada com enrofloxacina recuperou quase que totalmente a secreção láctea mas a eliminação de M. bovis persistiu por toda lactação. Esta vaca apresentou cura bacteriológica na lactação seguinte. O descarte dos animais positivos, monitora-mento bacteriológico e a aplicação correta das medidas de prevenção para as mastites contagiosas controlaram a disseminação de M. bovis nos rebanhos.In this study 713 cows were examined. The animals were from three dairy farms in northern Paraná and the southwest of the State of São Paulo. From these cows, 137 had mastitis. On the three farms, 8 cows (1.12% with Mycoplasma bovis mastitis were detected. Four were treated with tylosin and oxytetracyclin and three with enrofloxacin. There was no response to the treatments, and these animals were culled during the lactation period. One cow treated with enrofloxacin almost totally recovered milk production, but elimination of M. bovis continued during the lactation, and there was no bacteriological cure. This cow had a normal milk production in the next lactation period, without elimination of M. bovis. Culling of positive animals, the bacteriological study and correct application of preventive practices for contagious mastitis controlled the dissemination of M. bovis to other animals.

  13. Significant Association of Streptococcus bovis with Malignant Gastrointestinal Diseases

    Directory of Open Access Journals (Sweden)

    Salah Shanan

    2011-01-01

    Full Text Available Streptococcus bovis is a Gram-positive bacterium causing serious human infections, including endocarditis and bacteremia, and is usually associated with underlying disease. The aims of the current study were to compare prevalence of the bacterium associated with malignant and nonmalignant gastrointestinal diseases and to determine the susceptibility of the isolated strains to different antimicrobial agents. The result showed that the prevalence of S. bovis in stool specimens from patients with malignant or with nonmalignant gastrointestinal diseases was statistically significant. This result may support the idea that there is correlation between S. bovis and the malignant gastrointestinal diseases.

  14. Genetic variations among Mycoplasma bovis strains isolated from Danish cattle

    DEFF Research Database (Denmark)

    Kusiluka, L.J.M.; Kokotovic, Branko; Ojeniyi, B.

    2000-01-01

    The genetic heterogeneity of Mycoplasma bovis strains isolated in Denmark over a 17-year period was investigated. Forty-two field strains isolated from different geographic locations and specimens, including strains from 21 herds involved in two outbreaks of M. bovis-induced mastitis, and the type...... strain of M. bovis (PG45(T)) were assayed for variations in the BglII and MfeI restriction sites in the chromosomal DNA by using the amplified fragment length polymorphism (AFLP) fingerprinting technique. The obtained genomic fingerprints consisted of 62-68 AFLP fragments in the size range of 50-500 bp...... 2 years apart; showed indistinguishable AFLP patterns. More genetic diversify was observed among the recent strains. The similarity of the genotypes of the field strains to that of the M. bovis type strain (PG45(T)) was 97.7%. The results of this study have demonstrated a remarkable genomic...

  15. Cloning and sequencing of a Moraxella bovis pilin gene.

    OpenAIRE

    1985-01-01

    Moraxella bovis pili have been shown to play a major role in both infectivity and protective immunity of bovine infectious keratoconjunctivitis. Sonicated M. bovis DNA from the piliated strain EPP63 was inserted into the vector lambda gt11 with EcoRI linkers. Recombinant phage were screened with an oligonucleotide probe based on the amino-terminal portion of the DNA sequence of a Neisseria gonorrhoeae pilin gene. Two candidate phages produced a protein that comigrated with EPP63 beta pilin in...

  16. Experimental inoculation of wild turkeys (Meleagris gallopavo) with Mycobacterium bovis.

    Science.gov (United States)

    Clarke, K R; Fitzgerald, S D; Hattey, J A; Bolin, C A; Berry, D E; Church, S V; Reed, W M

    2006-03-01

    Although avian species are known to be susceptible to infection with Mycobacterium spp. organisms, much remains unknown about the susceptibility of birds to infection with M. bovis. The objective of this current study was to determine if wild turkeys (Meleagris gallopavo) can be infected with M. bovis when inoculated by the oral or intratracheal route. Six turkeys were orally inoculated and another six were inoculated via the trachea with a high dose of M. bovis, 1 x 10(5) CFU/ml. Six turkeys were sham-inoculated controls. Two turkeys from each treatment group were sacrificed on days 30, 60, and 90 postinoculation. There were no gross or microscopic lesions consistent with mycobacteriosis in the 23 inoculated turkeys over the 90-day duration of this study. Fecal cultures were also consistently negative for M. bovis when sampled before inoculation and on days 1, 30, and 60 postinoculation. Two intratracheally inoculated turkeys were positive for M. bovis in visceral tissues at 30 days postinoculation. However, this finding was only indicative of passive persistence of mycobacteria in the tissues and not of infection, as there were no attendant lesions or clinical compromise to support infection. Thus, it can be concluded that young wild turkeys are resistant to infection with M. bovis and, therefore, pose minimal threat as reservoir or spillover hosts for this organism.

  17. 严重联合免疫缺陷并发全身播散性卡介苗病一例%Disseminated infection with bacille Calmette-Guerin vaccine in an infant with severe combined immunodeficiency

    Institute of Scientific and Technical Information of China (English)

    刘雪芹; 孙君轶

    2007-01-01

    患儿男,8个月,因左下肢肿胀3个月、皮下结节2个月伴发热3d入院。患儿3个月前出现左下肢肿胀,皮肤发硬,当地医院诊为橡皮肿,未予治疗,2个月前患儿左上肢出现3个黄豆大小硬性皮下结节,2个月来皮下结节逐渐增多,散在分布于躯干及四肢,且体积逐渐增大至花生米大小,部分结节变软破溃流脓,曾多次在当地医院静脉输注抗生素治疗无效,入我院前3d起患儿中等度发热,发病以来患儿精神食欲差,体重增长缓慢。该患儿为第一胎第一产,足月顺产,生后第2天接种卡介苗,按时接种脊髓灰质炎疫苗及白百破疫苗,父母家系三代均无类似疾病史。查体:重病容,面色略苍白,发育营养稍差,左上臂卡介苗接种处见-1cm×1cm溃疡面,有脓性渗出,全身散在数十枚绿豆至花生米大小皮下结节,略高出皮面,质中偏硬,活动可,腹壁数个结节明显凸出皮面,结节内有波动感,部分破溃后有脓性液体流出,阴囊内可触及数个绿豆大小活动性结节,右侧睾丸下极一0.5cm×0.5cm硬性结节,全身浅表淋巴结未触及,心肺检查未见异常,肝肋下2.5cm,脾肋下3cm,质地中等,左下肢橡皮样肿胀。

  18. Immune Responses in Cattle Inoculated with Mycobacterium bovis, Mycobacterium tuberculosis, or Mycobacterium kansasii

    Science.gov (United States)

    Cattle were inoculated with Mycobacterium bovis, Mycobacterium tuberculosis, or Mycobacterium kansasii to compare antigen-specific immune responses to varied patterns of mycobacterial disease. Disease expression ranged from colonization with associated pathology (M. bovis), colonization without path...

  19. Characterization of Hemolysin of Moraxella bovis Using a Hemolysis-Neutralizing Monoclonal Antibody

    OpenAIRE

    2000-01-01

    A concentrated bacterial culture supernatant from the hemolytic Moraxella bovis strain UQV 148NF was used to immunize mice and generate monoclonal antibodies (MAbs). One, MAb G3/D7, neutralized the hemolytic activity of M. bovis and recognized a 94-kDa protein by Western blot analysis in hemolytic M. bovis strains representing each of the different fimbrial serogroups. Exposure of corneal epithelial cells to M. bovis concentrated culture supernatants demonstrated a role for an exotoxin in the...

  20. African 2, a clonal complex of Mycobacterium bovis epidemiologically important in East Africa.

    NARCIS (Netherlands)

    Berg, S.; Garcia-Pelayo, M.C.; Muller, B.; Hailu, E.; Asiimwe, B.; Kremer, K.; Dale, J.; Boniotti, M.B.; Rodriguez, S.; Hilty, M.; Rigouts, L.; Firdessa, R.; Machado, A.; Mucavele, C.; Ngandolo, B.N.; Bruchfeld, J.; Boschiroli, L.; Muller, A.; Sahraoui, N.; Pacciarini, M.; Cadmus, S.; Joloba, M.; Soolingen, D. van; Michel, A.L.; Djonne, B.; Aranaz, A.; Zinsstag, J.; Helden, P. van; Portaels, F.; Kazwala, R.; Kallenius, G.; Hewinson, R.G.; Aseffa, A.; Gordon, S.V.; Smith, N.H.

    2011-01-01

    We have identified a clonal complex of Mycobacterium bovis isolated at high frequency from cattle in Uganda, Burundi, Tanzania, and Ethiopia. We have named this related group of M. bovis strains the African 2 (Af2) clonal complex of M. bovis. Af2 strains are defined by a specific chromosomal deletio

  1. Identifikasi Cysticercus bovis pada Steak Daging Sapi di Rumah Makan Jalan Dr. Mansyur Medan Tahun 2013

    OpenAIRE

    Humaidi, Nur Qistina

    2015-01-01

    Cysticercus bovis merupakan larva cacing pita Taenia saginata. Larva ini dapat menyebabkan taeniasis saginata. Taeniasis terjadi apabila manusia mengkonsumsi daging mentah atau dimasak kurang matang yang telah terkontaminasi Cysticercus bovis. Penilitian ini bertujuan untuk mengidentifikasi Cysticercus bovis pada steak daging sapi di rumah makan Jalan Dr. Mansyur, Medan. Jenis penelitian deskriptif dengan desain cross sectional. Penelitian ini menggunakan metode total sam...

  2. Mycobacterium bovis infection in humans and cats in same household, Texas, USA, 2012

    Science.gov (United States)

    Mycobacterium bovis infection of cats is exceedingly rare in non-endemic regions for bovine tuberculosis. This case study describes the diagnosis and clinical management of pulmonary M. bovis infection in two indoor-housed cats and their association with at least one M. bovis-infected human in Texas...

  3. Recovery of Mycobacterium bovis from Soft Fresh Cheese Originating in Mexico▿

    OpenAIRE

    Harris, N. Beth; Payeur, Janet; Bravo, Doris; Osorio, Ruben; Stuber, Tod; Farrell, David; Paulson, Debra; Treviso, Scarlett; Mikolon, Andrea; Rodriguez-Lainz, Alfonso; Cernek-Hoskins, Shannon; Rast, Robert; Ginsberg, Michele; Kinde, Hailu

    2006-01-01

    Recent outbreaks of human tuberculosis in the United States caused by Mycobacterium bovis have implicated cheese originating in Mexico as a source of these infections. A total of 203 samples of cheese originating in Mexico were cultured, and M. bovis was recovered from one specimen. Therefore, M. bovis can be recovered from cheese and may be a source of human infections.

  4. Mycobacterium bovis meningitis in young Nigerian-born male

    DEFF Research Database (Denmark)

    Faurholt-Jepsen, Daniel; Lillebæk, Troels; Nielsen, Ming-Yuan

    2014-01-01

    dairy products in Nigeria but had no known contact with larger mammals. Before the development of meningitis, the patient had several contacts with the health system due to fever and non-specific symptoms. Finally, upon hospital admission, the patient was diagnosed with M. tuberculosis complex......In Denmark, tuberculous meningitis is rare. Central nervous system (CNS) involvement with Mycobacterium bovis is even rarer and has only been seen three times since 1992. We present a case of M. bovis meningitis in a previously healthy young Nigerian-born male, who had been exposed to unpasteurized...... meningitis and treated empirically. After 13 days he was discharged without neurological sequelae. Later, the culture revealed M. bovis and treatment was adjusted accordingly....

  5. Mycobacterium bovis meningitis in young Nigerian-born male.

    Science.gov (United States)

    Faurholt-Jepsen, Daniel; Lillebaek, Troels; Nielsen, Ming-Yuan; Nielsen, Susanne Dam

    2014-10-01

    In Denmark, tuberculous meningitis is rare. Central nervous system (CNS) involvement with Mycobacterium bovis is even rarer and has only been seen three times since 1992. We present a case of M. bovis meningitis in a previously healthy young Nigerian-born male, who had been exposed to unpasteurized dairy products in Nigeria but had no known contact with larger mammals. Before the development of meningitis, the patient had several contacts with the health system due to fever and non-specific symptoms. Finally, upon hospital admission, the patient was diagnosed with M. tuberculosis complex meningitis and treated empirically. After 13 days he was discharged without neurological sequelae. Later, the culture revealed M. bovis and treatment was adjusted accordingly.

  6. In vitro adherence of Moraxella bovis to intact corneal epithelium.

    Science.gov (United States)

    Jackman, S H; Rosenbusch, R F

    1984-09-01

    An in vitro assay is described using radiolabeled Moraxella bovis for studying adherence to intact bovine corneal epithelial surfaces. The assay was optimized for time (45 min) and for the ratio of epithelial cells to bacteria (1:1000) that demonstrated a significant difference in adherence between M. bovis strain 118F, a piliated organism and a nonpiliated variant, strain 118F/4-2. Adherence of these organisms correlated with previous pathogenicity studies involving experimental infection of calves. Scanning electron microscopy of tissues treated in the assay revealed a predilection of M. bovis for dark epithelial cells and for association with depressions in the tissue surface. This assay technique is discussed in comparison with other in vitro adherence assay methods.

  7. Adherence of Moraxella bovis to cell cultures of bovine origin.

    Science.gov (United States)

    Annuar, B O; Wilcox, G E

    1985-09-01

    The adherence of five strains of Moraxella bovis to cell cultures was investigated. M bovis adhered to cultures of bovine corneal epithelial and Madin-Darby bovine kidney cells but not to cell types of non-bovine origin. Both piliated and unpiliated strains adhered but piliated strains adhered to a greater extent than unpiliated strains. Antiserum against pili of one strain inhibited adherence of piliated strains but caused only slight inhibition of adherence to the unpiliated strains. Treatment of bacteria with magnesium chloride caused detachment of pili from the bacterial cell and markedly inhibited adherence of piliated strains but caused only slight inhibition of adherence by the unpiliated strains. The results suggested that adhesion of piliated strains to cell cultures was mediated via pili but that adhesins other than pili may be involved in the attachment of unpiliated strains of M bovis to cells.

  8. Blocking Babesia bovis vaccine reactions of dairy cattle in milk

    Directory of Open Access Journals (Sweden)

    Michael P. Combrink

    2012-12-01

    Full Text Available The use of 1.16 mg/kg (one third of the recommended dose of diminazene aceturate, administered indiscriminately to cattle on day seven of the unfrozen Babesia bovis and Babesia bigemina bivalent live blood vaccine reaction, was an infection and block treatment method of immunisation used successfully with no known adverse effect on the parasites or the development of protective immunity. Continuing with this practice after replacement of the unfrozen vaccine with deep-frozen monovalent B. bovis and B. bigemina live blood vaccines resulted in reports of vaccine failure. Laboratory investigation indicated the harmful effect of block treatment in preventing the development of durable immunity against B. bigemina as opposed to the much lesser effect it had on B. bovis. Consequently the practice was no longer recommended. A B. bovis vaccination attempt aimed at controlling the disease of dairy cows in milk (n = 30 resulted in 20% fatalities during the expected vaccine reaction period. The practice of block treating B. bovis was therefore reinvestigated, this time in a field trial using dairy cattle in milk (n = 11. Using 0.88 mg/kg (one quarter of the recommended dose of diminazene administered on day 12 of the B. bovis vaccine reaction resulted in only two animals (n = 5 testing ≥ 1/80 positive with the indirect fluorescent antibody test (IFAT although parasites could be demonstrated in three. In the untreated control group, by contrast, five of the vaccinated animals (n = 6 tested ≥ 1/80 positive with IFAT and parasites could be demonstrated in all. The unsatisfactory outcome obtained in this study, combined with that of the earlier investigation, indicated that there are more factors that influence successful vaccination than previously considered. It is therefore concluded that block treatment of the live frozen South African cattle babesiosis vaccines reactions is not recommended.

  9. Streptococcus bovis endocarditis and colon cancer: myth or reality? A case report and literature review.

    Science.gov (United States)

    Galdy, Salvatore; Nastasi, Giuseppe

    2012-12-05

    A relationship between infective endocarditis and colon cancer was established in 1950, and Streptococcus bovis was successfully isolated in 1970. However, this association and its pathogenesis still remain unclear. In this paper, we describe the clinical case of a patient with a history of colon cancer and infective endocarditis caused by Streptococcus bovis. The role of S bovis as an aetiological agent in the development of colon cancer is intriguing but uncertain. S bovis infection should be considered a silent sign of gastrointestinal malignancy or hepatic disease. We believe that in order to demonstrate the presence of colon cancer, all patients with S bovis infection require an endoscopic investigation of the colon.

  10. T-Cell mRNA Expression in Response to Mycobacterium bovis BCG Vaccination and Mycobacterium bovis Infection of White-Tailed Deer▿

    OpenAIRE

    Tyler C Thacker; Palmer, Mitchell V.; Waters, W. Ray

    2009-01-01

    Understanding immune responses of white-tailed deer (WTD) to infection with Mycobacterium bovis provides insight into mechanisms of pathogen control and may provide clues to development of effective vaccine strategies. WTD were vaccinated with either M. bovis BCG strain Pasteur or BCG strain Danish. Both vaccinees and unvaccinated controls were subsequently inoculated with virulent M. bovis via the intratonsillar route. Real-time PCR was used to assess T-cell mRNA expression in peripheral blo...

  11. Bacillus probiotics.

    Science.gov (United States)

    Cutting, Simon M

    2011-04-01

    Bacterial spore formers are being used as probiotic supplements for use in animal feeds, for human dietary supplements as well as in registered medicines. Their heat stability and ability to survive the gastric barrier makes them attractive as food additives and this use is now being taken forward. While often considered soil organisms this conception is misplaced and Bacilli should be considered as gut commensals. This review summarises the current use of Bacillus species as probiotics, their safety, mode of action as well as their commercial applications.

  12. Comparative Proteomic Profiling of Mycobacterium bovis and BCG Vaccine Strains

    KAUST Repository

    Gao, Ge

    2013-09-01

    BCG is the only licensed human vaccine currently available against TB. Derived from a virulent strain of M. bovis, the vaccine was thought to have struck a balance between reduced virulence and preserved immunogenicity. Nowadays, BCG vaccine strains used in different countries and vaccination programs show clear variations in their genomes and immune protective properties. The aim of this study was to characterize the proteomic profile on Mycobacterium bovis and five BCG strains Pasteur, Tokyo, Danish, Phipps and Birkhaug by Tandem Mass Tag® (TMT®)-labeling quantitative proteomic approach. In total, 420 proteins were identified and 377 of them were quantitated for their relative abundance. We reported the number and relationship of differential expressed proteins in BCG strains compared to M. bovis and investigated their functions by bioinformatics analysis. Several interesting up-regulated and down-regulated protein targets were found. The identified proteins and their quantitative expression profiles provide a basis for further understanding of the cellular biology of M. bovis and BCG vaccine strains, and hopefully would assist in the design of better anti-TB vaccine and drugs.

  13. 9 CFR 113.409 - Tuberculin-PPD Bovis, Intradermic.

    Science.gov (United States)

    2010-01-01

    ... shall not be released. (a) Purity test. Each serial shall be tested for viable bacteria and fungi as... the average response obtained on M. bovis sensitized guinea pigs. (8) Validity of bioassay. The bioassay test results obtained on serials tested concurrently in a single test series are valid if...

  14. Abortion in laboratory animals induced by Moraxella bovis.

    Science.gov (United States)

    Norman, J O; Elissalde, M H

    1979-05-01

    Pregnant mice, guinea pigs, rats, and rabbits responded to injections of Moraxella bovis strain EPP-63(300) with abortion, death, embryo resorption, and production of small litters. The nature of response appeared to depend primarily on the number of viable cells injected and to some extent on the species of animal and stage of pregnancy. Intraperitoneal injection of mice with 3 X 10(5) viable cells induced 100% abortion and no deaths. Embryo resorption and smaller litters were induced with injection of lower doses of M. bovis. None of the rats used in this study aborted; however, resorption rates were higher in rats than mice. Bacteria-free filtrates prepared from aqueous extracts of blood agar on which M. bovis had been grown induced abortion in mice and guinea pigs, at a rate similar to that caused by viable cells. The lyophilized filtrate could be diluted to produce 100% abortion with no death of injected mice. Abortion-inducing, nonlethal doses of M. bovis and lyophilized filtrates appeared to have no effect on nonpregnant female or male mice.

  15. The life and legacy of William T. Bovie.

    Science.gov (United States)

    Carter, Preston L

    2013-05-01

    This Historian's Address, presented at the North Pacific Surgical Association 2012 meeting, held in Spokane, Washington, on November 9, 2012, briefly reviews the life and surgical contributions of the inventor William T. Bovie and his collaboration with Dr Harvey Cushing, which led to the widespread acceptance of surgical electrocautery for dissection and hemostasis.

  16. First description of Bartonella bovis in cattle herds in Israel.

    Science.gov (United States)

    Rudoler, Nir; Rasis, Michal; Sharir, Benny; Novikov, Anna; Shapira, Gregory; Giladi, Michael

    2014-09-17

    Bartonella bovis has been described in beef and dairy cattle worldwide, however the reported prevalence rates are inconsistent, with large variability across studies (0-89%). This study describes the first isolation and characterization of B. bovis among cattle herds in the Middle East. Blood samples from two beef cattle herds (each sampled thrice) and one dairy herd (sampled twice) in Israel were collected during a 16-months period. Overall, 71 of 95 blood samples (75%) grew Bartonella sp., with prevalence of 78% and 59% in beef and dairy cattle, respectively. High level bacteremia (≥100,000 colony forming units/mL) was detected in 25 specimens (26%). Such high-level bacteremia has never been reported in cattle. Two dairy cows and one beef cow remained bacteremic when tested 60 or 120 days apart, respectively, suggesting that cattle may have persistent bacteremia. One third of animals were infested with ticks. Sequence analysis of a gltA fragment of 32 bacterial isolates from 32 animals revealed 100% homology to B. bovis. Species identification was confirmed by sequence analysis of the rpoB gene. Phylogenetic analysis based on the concatenated sequences of gltA and rpoB demonstrated that the isolates described herein form a monophyletic group with B. bovis strains originating from cattle worldwide. Taken together, the high prevalence of bacteremia, including high-level bacteremia, in beef and dairy cattle, the potential to develop prolonged bacteremia, the exposure of cattle to arthropod vectors, and proximity of infected animals to humans, make B. bovis a potential zoonotic agent.

  17. Cloning and sequencing of a Moraxella bovis pilin gene.

    Science.gov (United States)

    Marrs, C F; Schoolnik, G; Koomey, J M; Hardy, J; Rothbard, J; Falkow, S

    1985-07-01

    Moraxella bovis pili have been shown to play a major role in both infectivity and protective immunity of bovine infectious keratoconjunctivitis. Sonicated M. bovis DNA from the piliated strain EPP63 was inserted into the vector lambda gt11 with EcoRI linkers. Recombinant phage were screened with an oligonucleotide probe based on the amino-terminal portion of the DNA sequence of a Neisseria gonorrhoeae pilin gene. Two candidate phages produced a protein that comigrated with EPP63 beta pilin in sodium dodecyl sulfate-polyacrylamide gels and bound anti-pilus antisera. The 1.9-kilobase insert from one of these, lambda gt11M182, was subcloned in both orientations into pBR322, forming the plasmids pMxB7 and pMxB9, both of which produced beta pilin, as did pMxB12, a HindIII deletion derivative of pMxB7. In HB101(pMxB12), the M. bovis pilin protein was shown to be primarily localized in the inner membrane. The entire 939-base-pair insert of pMxB12 was sequenced, revealing a ribosome binding site just upstream of the coding region and an AT-rich region further upstream containing some potential RNA polymerase recognition sites. The translation of the sequence predicts a six-amino-acid leader sequence preceding the phenylalanine that begins the mature protein. Codon usage analysis of the M. bovis beta pilin gene revealed greater use of the CUA codon for leucine than usual for a well-expressed Escherichia coli gene. Comparisons of the M. bovis EPP63 beta pilin protein sequence with other pilin gene sequences are presented.

  18. Taurine as a marker for the identification of natural Calculus Bovis and its substitutes.

    Science.gov (United States)

    Shimada, Kayoko; Azuma, Yuko; Kawase, Masaya; Takahashi, Toshiharu; Schaffer, Stephen W; Takahashi, Kyoko

    2013-01-01

    Calculus Bovis (C. Bovis) is a commonly used animal-derived therapeutic preparation. To meet the increasing clinical demand for the preparation, two artificial substitutes for Bos Taurus have been introduced in China: artificial C. Bovis and in vitro cultured C. Bovis. However, information on their efficacy and safety is inadequate. Therefore, we investigated the biological differences between the commonly used natural preparation and its two substitutes, with the aim of not only identifying the differences but also providing a procedure to distinguish between the different preparations.In the study, we prepared 9 natural C. Bovis, 2 artificial C. Bovis, and 2 in vitro cultured C. Bovis preparations for evaluation. Differences were noted between the three preparations relative to their effect on viability of cardiac fibroblasts from 1-day-old Wistar rats. Although natural C. Bovis had no effect on cell viability, 1-h treatment of the cells with 0.25 mg/ml of the substitutes significantly reduced cell viability, as detected by the MTS assay. Based on liquid chromatography and inductively coupled plasma mass spectrometry, the preparations also differed in composition. Indeed, the substitutes contained more taurine, cholic acid, iron, magnesium, and calcium than the natural preparations. They also differed spectroscopically.The present results reveal significant biological differences between natural C. Bovis and two of its substitutes. Since the substitutes appear to contain more taurine, cholic acid, and elements, these constituents may serve as markers to distinguish between natural C. Bovis and its substitutes.

  19. Multiplex-PCR for differentiation of Mycobacterium bovis from Mycobacterium tuberculosis complex.

    Science.gov (United States)

    Spositto, F L E; Campanerut, P A Z; Ghiraldi, L D; Leite, C Q F; Hirata, M H; Hirata, R D C; Siqueira, V L D; Cardoso, R Fressatti

    2014-01-01

    We evaluated a multiplex-PCR to differentiate Mycobacterium bovis from M. tuberculosis Complex (MTC) by one step amplification based on simultaneous detection of pncA 169 C > G change in M. bovis and the IS6110 present in MTC species. Our findings showed the proposed multiplex-PCR is a very useful tool for complementation in differentiating M. bovis from other cultured MTC species.

  20. Mycobacterium bovis infection in a wild sow (Sus scrofa): first case in Korea.

    Science.gov (United States)

    Ku, Bok Kyung; Jeon, Bo-Young; Kim, Jae Myung; Jang, Young-Boo; Jang, Yunho; Yu, So Yoon; Kim, Jiro; Moon, Oun Kyung; Jung, Suk Chan; Lee, Min Kwon; Jeong, Tae Nam

    2016-09-30

    Mycobacterium (M.) bovis causes tuberculosis and has a broad host range, including humans, livestock, and wild animals. M. bovis infection of wild boar has been reported in several European countries. We report here the first case of M. bovis infection in a domesticated wild sow in Korea. Granulomatous and necrotizing lesions with small numbers of acid-fast bacilli were observed in nodules of the lung of wild sow. Furthermore, the M. bovis isolate from the wild sow had spoligotype SB0140 and a novel MIRU-VNTR allelic profile, which is not found in cattle and deer in Korea.

  1. An observational study of Corynebacterium bovis in selected Ontario dairy herds.

    OpenAIRE

    1983-01-01

    An observational study of Corynebacterium bovis was conducted in 74 Ontario dairy herds. The levels of infection with C. bovis were 19.9, 36.2 and 85.6% at the quarter, cow and herd level, respectively. Teat disinfection was found to be the variable best able to distinguish between herds with a high or low C. bovis quarter infection rate. Mean total milk somatic cell counts for 1103 quarters and 107 cows infected with only C. bovis ranged between 150,000 and 200,000/mL and were significantly ...

  2. Mycobacterium bovis in Swine: Spoligotyping of Isolates from Argentina

    Directory of Open Access Journals (Sweden)

    Soledad Barandiaran

    2011-01-01

    Full Text Available A total of 143 Mycobacterium bovis isolates of pigs, from the most productive swine area in Argentina, were typed by spoligotyping. Twenty-two different spoligotypes were identified, and 133 (93% isolates were grouped into 12 clusters. One of them, designed SB0140, was the most frequent because it held 83 (58% isolates. This spoligotype also grouped 362 (43% out of 841 isolates from previously typed cattle and, thus, constitutes the most frequent in our country. In addition, 135 (94% isolates revealed spoligotypes identical to those of cattle, showing an epidemiological link. On the other hand, there were seven novel spoligotypes, six of which were also unique since they had only one isolate each. This study aimed to identify the spoligotypes of M. bovis isolated from pigs to contribute to a better understanding of the distribution of bovine tuberculosis in the main productive area of Argentina.

  3. Importance of differentiating Mycobaterium bovis in tuberculous meningitis

    Directory of Open Access Journals (Sweden)

    Alejandra Gonzàlez-Duarte

    2011-08-01

    Full Text Available The aim of the article is to describe the principal findings among patients with M.tuberculosis and M. bovis CNS infection. Mycoba - cterium tuberculosis is one of the most common infectious agents that cause death and neurological sequelae around the world. Most of the complications of CNS TB can be attributed to a delay in the diagnosis. Unfortu nately, there are no specific diagnostic tools to support an early diagnosis. Other prognostic factors different from delay in treatment have not been identified. Clinical, radiological and laboratory characteristics were analyzed retrospectively from the medical files of all the patients admitted with the diagnoses of tuberculosis. Of 215 patients admitted with systemic tuberculosis, 64 (30% had a neurological infection. Positive cultures were found in 54 (84% cases, 18 (33% in the CSF and the rest in other fluids or tissues. Adenosin deaminase (ADA enzyme determination was more sensitive than M. tuberculosis PCR in the CSF for supporting an early diagnosis. In addition to a later clinical stage and treatment lag, positive CSF cultures (P=0.001 and the presence of M. bovis (P=0.020 were prognostic factors for a worse outcome. Neither older age, the presence of tuberculomas versus meningeal enhancement, or HIV co-infection, was associated to a worse prognosis. The isolation of M. bovis subspecies was more common that previously reported, and it was associated to the development of parenchymal lesions (P=0.032 when compared to M. tuberculosis. In this study, positive CSF cultures for M. tuberculosis and further identifying M. bovis species were additional prognostic factors for worse outcome. Positive cultures in systemic fluids other than CSF, even when the patient had no obvious systemic manifestations, and ADA determination in the CSF were noteworthy diagnostic tools for the diagnosis.

  4. Quantification of Moraxella bovis haemagglutinating adhesins with monoclonal antibodies.

    Science.gov (United States)

    Gil-Turnes, C; Aleixo, J A

    1991-08-01

    Six monoclonal antibodies (MAbs) against Moraxella bovis GF 9 were used to quantify haemagglutinating adhesins of 16 strains of this organism. The amount of each MAb necessary to inhibit one haemagglutinating unit of each strain varied between 4 and 0.007 times that required by strain GF 9. Five strains reacted with six MAbs, one with five, two with four, one with three, two with two and three with none. The procedures used enabled to detect dominant strains candidates for vaccines.

  5. Purification, characterization, and pathogenicity of Moraxella bovis pili

    OpenAIRE

    1988-01-01

    Pilins composed of the alpha or beta pilins of Moraxella bovis strain Epp63 were purified, subjected to chemical or enzymatic cleavage, and the resulting fragments sequenced by automated Edman degradation. alpha Pilin was found to be a 155-amino-acid polypeptide with a single intramolecular disulfide bridge. The beta pilin amino acid sequence substantiated the previously reported structure derived from the beta pilin gene DNA sequence, and indicated that the alpha and beta pilins of this stra...

  6. Mycobacterium bovis: realities and challenges for the veterinary biopharmaceutical industry

    Directory of Open Access Journals (Sweden)

    Aníbal Domínguez Odio

    2016-01-01

    Full Text Available Mycobacterium bovis is the main etiological agent of bovine tuberculosis, bacterial diseases of world distribution, chronicle, of easy transmission, debilitating, zoonotic and antropozoonotic that affects any organ and which can be presented without symptoms On this base, it was carried out a study with the objective of approaching the current state and the scientific-technological projections for the prevention and diagnosis of the bovine tuberculosis, caused by M. bovis. It was demonstrated that the 45.09% of the original articles on inmunoprophylaxis against bacteria, registered in the Scopus database and contextualised until principles of 2014, were focused toward M. bovis. In spite of the advances in molecular biology and the hopes deposited in the Ag85A, Rv0287, Rv0288, Rv0251c, MPB70, MPB83, ESAT-6 and CFP-10 molecules, jointly with their combinations, it will continue absent in the market an effective, safety and differentiating vaccine; as well as a robust DIVA diagnosis system. It can be concluded that in the next 5 years, an officially recognized vacinal formulation will continue absent and that the tuberculin test in spite of its weaknesses will continue being the main tool of surveillance.

  7. IFN-γ release assay: a diagnostic assistance tool of tuberculin skin test in pediatric tuberculosis in China

    Institute of Scientific and Technical Information of China (English)

    SUN Lin; SHEN A-dong; YAN Hui-min; HU Ying-hui; JIAO Wei-wei; GU Yi; XIAO Jing; LI Hui-min; JIAO An-xia; GUO Ya-jie

    2010-01-01

    Background Prompt diagnosis of Mycobacterium tuberculosis (MTB) infection is an essential step in tuberculosis control and elimination. However, it is often difficult to accurately diagnose pediatric tuberculosis (TB). The tuberculin test (TST) may have a low specificity because of cross-reactivity with antigens present in Mycobacterium bovis bacillus Calmette-Guerin (BCG) and other mycobacteria, especially in China with a predominantly BCG-vaccinated population.Early-secreted antigenic target 6-kDa protein (ESAT-6) and culture filtrate protein 10 (CFP-10), stand out as suitable antigens that induce an interferon-gamma (IFN-γ) secreting, T-cell-mediated immune response to infection. While,considered the higher costs and complexity of the IFN-γ release assay (TSPOT), we aimed to evaluate the TSPOT and TST test in the clinical diagnosis of pediatric tuberculosis and to establish a diagnostic process suitable for China.Methods The sensitivity and specificity of the assay were evaluated in total seventy four children with active tuberculosis and fifty one nontuberculous children with other disease, and then the results were compared with TST.Logistic regression models were used to identify variables that were associated with positive results for each assay. The independent variables included sex, age, birth place, vaccination history, close contract with an active TB patient.Results The sensitivity of TSPOT was higher than TST in active TB children with or without BCG vaccination, as well as in children with culture-confirmed TB. But the difference was not significant statistically. Combining results of the TSPOT and TST improved the sensitivity to 94.6%. Agreement of the TST and TSPOT was low (77.0%, k=0.203) in active TB patients. The difference in specificity between TSPOT and TST test was statistically significant (94.1% vs.70.6%, P=0.006). Specificity of the two tests in patients without prior BCG vaccination history was similar (80.0% vs.60.0%). The concordance

  8. The role of the mycobacterial DNA-binding protein 1 (MDP1 from Mycobacterium bovis BCG in host cell interaction

    Directory of Open Access Journals (Sweden)

    Kunisch Ralph

    2012-08-01

    Full Text Available Abstract Background Mycobacterium tuberculosis differs from most pathogens in its ability to multiply inside monocytes and to persist during long periods of time within granuloma in a status of latency. A class of proteins called mycobacterial histone-like proteins has been associated with regulation of replication and latency, but their precise role in the infection process has yet to be uncovered. Our study aimed at defining the impact of the histone-like protein MDP1 from M. bovis BCG (mycobacterial DNA-binding protein 1, corresponding to Rv2986c from M. tuberculosis on early steps of infection. Results Previously, a BCG (Bacillus Calmette Guérin strain had been generated by antisense-technique exhibiting reduced MDP1 expression. This strain was now used to analyse the impact of reduced amount of MDP1 on the interaction with human blood monocytes, macrophage lines and PBMC (peripheral blood mononuclear cells. MDP1 was revealed to be required for growth at acidic pH and for intracellular replication in human blood monocytes. Down-regulation of MDP1 resulted in reduced secretion of the cytokine IL-1β by infected human PBMC. In addition, a reduction of MDP1 expression had a major impact on the formation of fused multi-nucleated macrophages. In monocyte preparations from human blood as well as in human and mouse macrophage cell lines, both the percentage of multi-nucleated cells and the number of nuclei per cell were much enhanced when the monocytes were infected with BCG expressing less MDP1. Conclusion MDP1 from M. bovis BCG affects the growth at acidic pH and the intracellular replication in human monocytes. It furthermore affects cytokine secretion by host cells, and the formation of fused multi-nucleated macrophages. Our results suggest an important role of MDP1 in persistent infection.

  9. Coexisting Bacillus Calmette-Guérin-Induced Lupus Vulgaris Involving the Vaccination Site and Lichen Scrofulosorum in an Immunocompetent Boy.

    Science.gov (United States)

    Angoori, Gnaneshwar Rao

    2016-09-01

    The coexistence of Bacillus Calmette-Guérin (BCG)-induced lupus vulgaris involving the site of vaccination with lichen scrofulosorum is rare. Herein we report a 3-year-old boy who presented with lupus vulgaris at the vaccination site 3 weeks after neonatal BCG vaccination followed by the development of lichen scrofulosorum approximately 2.5 years later. Characteristic clinical morphology, typical histopathology, and positive DNA polymerase chain reaction for Mycobacterium bovis confirmed the clinical diagnosis.

  10. Polymorphisms of twenty regulatory proteins between Mycobacterium tuberculosis and Mycobacterium bovis

    Science.gov (United States)

    Mycobacterium tuberculosis and Mycobacterium bovis are responsible for tuberculosis in humans or animals, respectively. Both species are closely related and belong to the Mycobacterium tuberculosis complex (MTC). M. tuberculosis is the most ancient species from which M. bovis and the other members o...

  11. Is colonoscopy necessary in cases of infection by Streptococcus bovis biotype II?

    NARCIS (Netherlands)

    Corredoira, J.C.; Alonso, M.P.; Garcia-Pais, M.J.; Rabunal, R.; Garcia-Garrote, F.; Lopez-Roses, L.; Lancho, A.; Coira, A.; Pita, J.; Velasco, D.; Lopez-Alvarez, M.J.; Tjalsma, H.; Varela, J.

    2014-01-01

    The association of colorectal neoplasia (CRN) with Streptococcus bovis biotype I (SBI) infection is well recognized. However, this is not the case for Streptococcus bovis biotype II (SBII). We conducted this study in order to analyze the relationship between SBII and CRN. We analyzed all cases of ba

  12. In situ cytokine expression in pulmonary granulomas of cattle experimentally infected by aerosolized Mycobacterium bovis

    Science.gov (United States)

    Mycobacterium bovis is the cause of tuberculosis in most animal species, including cattle and is a serious zoonotic pathogen. In humans, M. bovis infection can result in disease clinically indistinguishable from that caused by Mycobacterium tuberculosis, the cause of most tuberculosis in humans. Reg...

  13. Multilocus sequence typing of Mycoplasma bovis reveals host-specific genotypes in cattle versus bison

    Science.gov (United States)

    Mycoplasma bovis is a primary agent of mastitis, pneumonia and arthritis in cattle and is the bacterium isolated most frequently from the polymicrobial syndrome known as bovine respiratory disease complex (BRDC). Recently, M. bovis has emerged as a significant health problem in bison, causing necro...

  14. A multilocus sequence typing method and curated database for Mycoplasma bovis

    Science.gov (United States)

    Mycoplasma bovis is a primary agent of mastitis, pneumonia and arthritis in cattle and is the bacterium isolated most frequently from the polymicrobial syndrome known as bovine respiratory disease complex (BRDC). Recently, M. bovis has emerged as a significant problem in bison, causing necrotic pha...

  15. Molecular characterization of Mycobacterium bovis strains isolated from cattle slaughtered at two abattoirs in Algeria

    Directory of Open Access Journals (Sweden)

    Ouzrout Rachid

    2009-01-01

    Full Text Available Abstract Background Bovine Tuberculosis is prevalent in Algeria despite governmental attempts to control the disease. The objective of this study was to conduct, for the first time, molecular characterization of a population sample of Mycobacterium bovis strains isolated from slaughter cattle in Algeria. Between August and November 2007, 7250 animals were consecutively screened at the abattoirs of Algiers and Blida. In 260 animals, gross visible granulomatous lesions were detected and put into culture. Bacterial isolates were subsequently analysed by molecular methods. Results Altogether, 101 bacterial strains from 100 animals were subjected to molecular characterization. M. bovis was isolated from 88 animals. Other bacteria isolated included one strain of M. caprae, four Rhodococcus equi strains, three Non-tuberculous Mycobacteria (NTM and five strains of other bacterial species. The M. bovis strains isolated showed 22 different spoligotype patterns; four of them had not been previously reported. The majority of M. bovis strains (89% showed spoligotype patterns that were previously observed in strains from European cattle. Variable Number of Tandem Repeat (VNTR typing supported a link between M. bovis strains from Algeria and France. One spoligotype pattern has also been shown to be frequent in M. bovis strains from Mali although the VNTR pattern of the Algerian strains differed from the Malian strains. Conclusion M. bovis infections account for a high amount of granulomatous lesions detected in Algerian slaughter cattle during standard meat inspection at Algiers and Blida abattoir. Molecular typing results suggested a link between Algerian and European strains of M. bovis.

  16. Effect of skin test on serum antibody responses to Mycobacterium bovis infection in cattle

    Science.gov (United States)

    Recently, several serologic tests designed to detect immunodominant antibodies to M. bovis antigens (e.g., MPB83, MPB70, ESAT-6, and CFP10) have emerged for potential use with samples from cattle. Of these, a commercial ELISA to MPB83/MPB70 (M. bovis antibody ELISA) has gained approval for use in ca...

  17. Q pili enhance the attachment of Moraxella bovis to bovine corneas in vitro.

    Science.gov (United States)

    Ruehl, W W; Marrs, C; Beard, M K; Shokooki, V; Hinojoza, J R; Banks, S; Bieber, D; Mattick, J S

    1993-01-01

    Moraxella bovis, the causative agent of infectious bovine keratoconjunctivitis, exhibits several virulence factors, including pili, haemolysin, leukotoxin, and proteases. The pili are filamentous appendages which mediate bacterial adherence. Prior studies have shown that Q-piliated M. bovis Epp63 are more infectious and more pathogenic than I-piliated and non-piliated isogenic variants, suggesting that Q pili per se, or traits associated with Q-pilin expression, promote the early association of Q-piliated bacteria with bovine corneal tissue. In order to better evaluate the role of Q pili in M. bovis attachment, several M. bovis strains and a recombinant P. aeruginosa strain which elaborates M. bovis Q pili but not P. aeruginosa PAK pili, were evaluated using an in vitro corneal attachment assay. For each strain tested, piliated organisms attached better than non-piliated bacteria. M. bovis Epp63 Q-piliated bacteria adhered better than either the I-piliated or non-piliated isogenic variants. Finally, recombinant P. aeruginosa organisms elaborating M. bovis Q pili adhered better than the parent P. aeruginosa strain which did not produce M. bovis pili. These results indicate that the presence of pili, especially Q pili, enhances the attachment of bacteria to bovine cornea in vitro.

  18. Infectious bovine keratoconjunctivitis: effects of vaccination on Moraxella bovis carrier state in cattle.

    Science.gov (United States)

    Pugh, G W; McDonald, T J; Kopecky, K E

    1980-02-01

    A study was conducted to determine whether vaccination of cattle while they were undergoing an acute episode of infectious bovine keratoconjunctivitis (IBK), would cause vaccinated cattle to abort Moraxella bovis infection sooner than nonvaccinated cattle. Fourteen calves were allotted into two groups of seven calves each, and the eyes of each calf were exposed to a virulent culture of M bovis. Twenty days after calves were infected and showing signs of IBK, seven calves were vaccinated with M bovis pilus vaccine made from the exposure strain. Ocular and nasal discharges were collected and examined for M bovis for 64 days. Most calves developed signs of IBK after exposure and all but one calf (nonvaccinated) developed ocular infection with M bovis. The mean number of days (33 and 33.3, respectively) of ocular infection was not significantly (P = 0.05) different in the vaccinated calves than in the nonvaccinated calves. The eyes of the calf that did not become infected with M bovis were infected with nearly a pure culture of Escherichia coli. This calf failed to develop ocular infection, but had M bovis in its nasal discharge throughout the study (64 days). Moraxella bovis was isolated from the nasal discharge of other calves only when there were concurrent ocular infections.

  19. Complete Genome Sequences of Field Isolates of Mycobacterium bovis and Mycobacterium caprae.

    Science.gov (United States)

    de la Fuente, José; Díez-Delgado, Iratxe; Contreras, Marinela; Vicente, Joaquín; Cabezas-Cruz, Alejandro; Manrique, Marina; Tobes, Raquel; López, Vladimir; Romero, Beatriz; Domínguez, Lucas; Garrido, Joseba M; Juste, Ramón; Gortazar, Christian

    2015-06-25

    Here we report the complete genome sequences of field isolates of Mycobacterium bovis and the related mycobacterial species, Mycobacterium caprae. The genomes of three M. bovis (MB1, MB3, MB4) and one M. caprae (MB2) field isolates with different virulence, prevalence, and host distribution phenotypes were sequenced.

  20. Molecular detection of natural Babesia bovis infection from water buffaloes (Bubalus bubalis) and crossbred cattle

    DEFF Research Database (Denmark)

    Mahmmod, Yasser

    2013-01-01

    Babesia bovis (B. bovis) is a major causative agent of bovine babesiosis, with a considerable worldwide impact. The objective of this study was to evaluate the usefulness of PCR assay and microscopical examination (ME) for detection of B. bovis in naturally infected and apparently healthy water...... were apparently healthy. Two blood samples were collected from each animal; one was used for preparation of Giemsa-stained smears for ME while the other sample was used for DNA extraction and PCR testing. Out of 34 cattle and buffaloes, ME identified 13 animals (38.2%) as infected by B. bovis whereas...... apparently healthy, 5 animals (26.3%) were identified as infected by ME meanwhile 15 animals (78.9%) were identified by PCR. In conclusion, our findings demonstrated that water buffalos are likely to have a natural tolerance to B. bovis pathogen and/or more likely to be persistent carriers which were...

  1. The first cases of Bartonella bovis infection in cattle from Central Europe.

    Science.gov (United States)

    Welc-Falęciak, Renata; Grono, Krzysztof

    2013-03-23

    Bartonella bovis was recently identified as a cause of bovine endocarditis, although Bartonella infections in natural hosts are usually asymptomatic. The disease is often misdiagnosed and is only discovered during the slaughtering process. In Europe B. bovis infections in cattle were reported only in France and Italy, nothing is known about the occurrence of B. bovis in cattle for the northern and eastern parts of Europe. The aim of our study was to search for Bartonella DNA in cattle in Central Europe (Poland) using three different loci (rpoB, ITS 16-23S rRNA, gltA). Our study resulted in the first detection of the asymptomatic B. bovis infection in 6.8% (12/177) of cattle in Central Europe. The potential role of B. bovis as a zoonotic agent for domestic animals and human diseases creates the need for further studies of these bacteria in natural and accidental hosts.

  2. Eye Irritation Test of Bovis Calculus Pharmacopuncture Solutions for Eye Drop

    Directory of Open Access Journals (Sweden)

    Hyeong-sik Seo

    2008-06-01

    Full Text Available Objective : This study was done to investigate the safety of Bovis Calculus pharmacopuncture solution manufactured with freezing dryness method to use eye drop. Methods : The eye irritation test of this material was performed according to the Regulation of Korea Food & Drug Administration (2005. 10. 21, KFDA 2005-60. After Bovis Calculus pharmacopuncture solution was medicated in the left eye of the rabbits, the auther observed eye irritation of the cornea, iris, conjunctiva at 1, 2, 3, 4 & 7day. Results : 1. After Bovis Calculus pharmacopuncture solution was medicated in the left eye of the rabbits, there wasn’t physical problem at 9 rabbits. 2. After Bovis Calculus pharmacopuncture solutionwas medicated in the left eye of the rabbits, there wasn’t eye irritation of the cornea, iris, conjunctiva at 1, 2, 3, 4 & 7day. Conclusions : I suggested that Bovis Calculus pharmacopuncture solution didn’t induced eye irritation in rabbits.

  3. Identification of Mycobacterium bovis Isolates by a multiplex PCR Identificação de colônias isoladas de Mycobacterium bovis por PCR múltipla

    Directory of Open Access Journals (Sweden)

    Eduardo Eustáquio de Souza Figueiredo

    2009-06-01

    Full Text Available Isolates from suggestive bovine tuberculosis lesions were tested by a multiplex polymerase chain reaction (m-PCR targeting for RvD1Rv2031c and IS6110 sequences, specific for M. bovis and Mycobacterium tuberculosis complex respectively. The m-PCR successfully identified as M. bovis 88.24% of the isolates.Colônias isoladas a partir de lesões sugestivas de tuberculose bovina foram testadas pela reação múltipla em cadeia da polimerase, usando oligonucleotídeos direcionados para as seqüências genômicas RvD1Rv2031c e IS6110, específicas para M. bovis e para o complexo Mycobacterium tuberculosis, respectivamente. A m-PCR identificou, com sucesso, 88,24% das colônias isoladas como M. bovis.

  4. Cerebral ischemia caused by Streptococcus bovis aortic endocarditis: case report Isquemia cerebral causada por endocardite aórtica pelo Streptococcus bovis: relato de caso

    Directory of Open Access Journals (Sweden)

    Leopoldo Santos-Neto

    2005-09-01

    Full Text Available Cerebral ischemic processes associated with infective endocarditis caused by Streptococcus bovis are rare; only 2 cases having been reported. Here we report a case of a 50-year-old man with S. bovis endocarditis who presented signs of frontal, parietal and occipital lobe cerebral ischemia. This is the first case reported in which the presence of hemianopsia preceded the endocarditis diagnosis. Initially, the clinical manifestations suggested a systemic vasculitis. Later, vegetating lesions were identified in the aortic valve and S. bovis grew in blood cultures. Antibiotic use and aortic valve replacement eliminated the infection and ceased thromboembolic events. A videocolonoscopy examination revealed no mucosal lesions as a portal of entry in this case, although such lesions have been encountered in up to 70% of reported cases of S. bovis endocarditis.A associação de isquemia cerebral e endocardite por Streptococcus bovis é um evento raro, tendo sido publicados apenas 2 casos anteriormente. Nós relatamos o caso de um homem de 50 anos com endocardite por S. bovis que apresentou sinais isquêmicos nos lobos frontal, parietal e occipital. Este é o primeiro caso em que a hemianopsia precedeu o diagnóstico de endocardite. Inicialmente, o quadro foi confundido com vasculite. Posteriormente, foi confirmada a presença de vegetações na válvula aórtica e a hemocultura identificou S. bovis. Os eventos tromboembólicos foram controlados com o uso de antibióticos e a troca da válvula aórtica. Estudo videocolonoscópico não identificou nenhuma lesão, apesar de lesões colônicas serem descritas em até 70% dos casos de indivíduos com endocardite por S. bovis.

  5. Development of a specific DNA probe and PCR for the detection of Mycoplasma bovis.

    Science.gov (United States)

    Ghadersohi, A; Coelen, R J; Hirst, R G

    1997-05-01

    Mycoplasma bovis is responsible for several production diseases in cattle, including mastitis, arthritis, pneumonia, abortion and infertility. Current methodologies for detecting and identifying M. bovis are time consuming and difficult. Tests which rely on antigen or antibody detection have poor sensitivity and specificity. In this paper associated protocols for the development of a hybridization probe and PCR are described. A genomic library (SauIIIA digested) was prepared from M. bovis DNA (Colindale Reference Strain: NC10131:02) and cloned into pUC19. Colony hybridization, using a probe preparation made from purified M. bovis DNA, was used to identify colonies of interest. M. bovis DNA fragments were retrieved from recombinant plasmids by digestion with EcoRI and HindIII. This DNA was used to prepare randomly primed probes for dot blot hybridization analysis with immobilized DNA from M. bovis (two strains), M. dispar, M. agalactiae, M. bovigenitalium (two strains), M. ovipneumoniae, a Group 7 strain, M. arginini and bacteria belonging to different genera. Four probes were found to hybridize only with M. bovis and M. ovipneumoniae DNA, whereas one probe reacted with genomic DNA from only one of the two M. bovis strains. The level of sensitivity of the dot blot hybridization assay was 200 CFU (colony forming units)/mL. To enhance the sensitivity further, an M. bovis-specific PCR assay was developed. The primers were designed using sequences obtained from the probe DNA which discriminated M. bovis from all other Mycoplasma DNA tested. The minimum amount of target DNA that could be detected by the PCR assay was that isolated from 10-20 CFU/mL. The PCR assay was therefore 10 times more sensitive than dot blot hybridization.

  6. Increasing prevalence of Mycoplasma bovis in Danish cattle

    DEFF Research Database (Denmark)

    Kusiluka, L.J.M.; Ojeniyi, B.; Friis, N.F.

    2000-01-01

    A study on the prevalence of mycoplasmas in pneumonic bovine lungs was performed on material submitted for diagnostic pul poses at the Danish Veterinary Laboratory, Copenhagen. Among the 50 examined cases 43 (86.0%) were found to be infected with mycoplasmas. The predominant mycoplasmas were...... electrophoresis (PFGE) analysis of ii field isolates of M. bovis from 9 different farms revealed different profiles except for 2 isolates which were recovered from the same farm. Because mycoplasmas belonging to the M.mycoides cluster' were not encountered during this study; it appears that the Danish cattle...

  7. [In vitro antibiotic sensitivity of French strains of Mycoplasma bovis].

    Science.gov (United States)

    Poumarat, F; Martel, J L

    1989-01-01

    The in vitro activity of 15 antibiotics was tested with 30-90 Mycoplasma bovis representative strains of bovine lung pathology in France. The distribution of minimal inhibitory concentration (MIC) is homogeneous with low values for spectinomycin, lincomycin, tylosin, gentamicin and baytril, intermediate for chloramphenicol and neomycin, high for nalidixic acid, Flumequine and erythromycin. The MIC distribution is heterogeneous with intermediate values for spiramycin and tetracyclines, and high values for streptomycin. For the later antibiotics, the heterogeneity of the susceptibility suggests a mechanism of acquired resistance.

  8. Verrucous endocarditis associated with Streptococcus bovis in mink (Mustela vison)

    DEFF Research Database (Denmark)

    Pedersen, Karl; Jørgensen, J.C.; Dietz, Hans-Henrik

    2003-01-01

    Between 1998 and 2001, mortalities due to verrucous endocarditis were experienced at several mink farms. Gram-positive cocci were isolated from the endocardium of all the animals examined but not always from other internal organs. Almost all the isolates were identified as Streptococcus bovis...... and only a few isolates belonged to other Streptococcus species. Typing by pulsed-field gel electrophoresis of a selection of isolates revealed several patterns and several different clones. Attempts to reproduce disease by the injection of cultures of a field isolate into healthy mink failed....

  9. Optimization of modified Middlebrook 7H11 agar for isolation of Mycobacterium bovis from raw milk cheese.

    Science.gov (United States)

    Forgrave, R; Donaghy, J A; Fisher, A; Rowe, M T

    2014-10-01

    Reports have highlighted the absence of contemporary peer reviewed publications pertaining to Mycobacterium bovis culture from raw milk and cheese. By replicating traditional methods, cheese-making methodology and equipment were devised to produce Cheddar (n = 6) and Caerphilly (n = 3) artificially contaminated with M. bovis (three genotypes) under stringent laboratory-containment guidelines for handling hazardous microbiological material. Middlebrook 7H11, modified for M. bovis isolation, was assessed for capacity to enumerate M. bovis despite changing cheese microflora and prolonged M. bovis exposure to the cheese matrix using maturing cheese test portions (n = 63; up to 16 weeks). Malachite green (MG) containing media isolated M. bovis at significantly (P cheese types, Cheddar and Caerphilly. Significance and impact of the study: Following increased M. bovis infection of UK cattle, the risk posed to consumers from consumption of unpasteurized milk and dairy products has changed. Furthermore, published methods for the culture and molecular detection of M. bovis in raw milk products are limited. Cheese-making protocols and M. bovis culture media reported here provide tools for further investigation of M. bovis survival during all stages of cheese manufacture and could inform future assessment of the risk to consumers from M. bovis contamination of unpasteurized dairy products.

  10. African 2, a clonal complex of Mycobacterium bovis epidemiologically important in East Africa.

    Science.gov (United States)

    Berg, Stefan; Garcia-Pelayo, M Carmen; Müller, Borna; Hailu, Elena; Asiimwe, Benon; Kremer, Kristin; Dale, James; Boniotti, M Beatrice; Rodriguez, Sabrina; Hilty, Markus; Rigouts, Leen; Firdessa, Rebuma; Machado, Adelina; Mucavele, Custodia; Ngandolo, Bongo Nare Richard; Bruchfeld, Judith; Boschiroli, Laura; Müller, Annélle; Sahraoui, Naima; Pacciarini, Maria; Cadmus, Simeon; Joloba, Moses; van Soolingen, Dick; Michel, Anita L; Djønne, Berit; Aranaz, Alicia; Zinsstag, Jakob; van Helden, Paul; Portaels, Françoise; Kazwala, Rudovick; Källenius, Gunilla; Hewinson, R Glyn; Aseffa, Abraham; Gordon, Stephen V; Smith, Noel H

    2011-02-01

    We have identified a clonal complex of Mycobacterium bovis isolated at high frequency from cattle in Uganda, Burundi, Tanzania, and Ethiopia. We have named this related group of M. bovis strains the African 2 (Af2) clonal complex of M. bovis. Af2 strains are defined by a specific chromosomal deletion (RDAf2) and can be identified by the absence of spacers 3 to 7 in their spoligotype patterns. Deletion analysis of M. bovis isolates from Algeria, Mali, Chad, Nigeria, Cameroon, South Africa, and Mozambique did not identify any strains of the Af2 clonal complex, suggesting that this clonal complex of M. bovis is localized in East Africa. The specific spoligotype pattern of the Af2 clonal complex was rarely identified among isolates from outside Africa, and the few isolates that were found and tested were intact at the RDAf2 locus. We conclude that the Af2 clonal complex is localized to cattle in East Africa. We found that strains of the Af2 clonal complex of M. bovis have, in general, four or more copies of the insertion sequence IS6110, in contrast to the majority of M. bovis strains isolated from cattle, which are thought to carry only one or a few copies.

  11. African 2, a Clonal Complex of Mycobacterium bovis Epidemiologically Important in East Africa▿ †

    Science.gov (United States)

    Berg, Stefan; Garcia-Pelayo, M. Carmen; Müller, Borna; Hailu, Elena; Asiimwe, Benon; Kremer, Kristin; Dale, James; Boniotti, M. Beatrice; Rodriguez, Sabrina; Hilty, Markus; Rigouts, Leen; Firdessa, Rebuma; Machado, Adelina; Mucavele, Custodia; Ngandolo, Bongo Nare Richard; Bruchfeld, Judith; Boschiroli, Laura; Müller, Annélle; Sahraoui, Naima; Pacciarini, Maria; Cadmus, Simeon; Joloba, Moses; van Soolingen, Dick; Michel, Anita L.; Djønne, Berit; Aranaz, Alicia; Zinsstag, Jakob; van Helden, Paul; Portaels, Françoise; Kazwala, Rudovick; Källenius, Gunilla; Hewinson, R. Glyn; Aseffa, Abraham; Gordon, Stephen V.; Smith, Noel H.

    2011-01-01

    We have identified a clonal complex of Mycobacterium bovis isolated at high frequency from cattle in Uganda, Burundi, Tanzania, and Ethiopia. We have named this related group of M. bovis strains the African 2 (Af2) clonal complex of M. bovis. Af2 strains are defined by a specific chromosomal deletion (RDAf2) and can be identified by the absence of spacers 3 to 7 in their spoligotype patterns. Deletion analysis of M. bovis isolates from Algeria, Mali, Chad, Nigeria, Cameroon, South Africa, and Mozambique did not identify any strains of the Af2 clonal complex, suggesting that this clonal complex of M. bovis is localized in East Africa. The specific spoligotype pattern of the Af2 clonal complex was rarely identified among isolates from outside Africa, and the few isolates that were found and tested were intact at the RDAf2 locus. We conclude that the Af2 clonal complex is localized to cattle in East Africa. We found that strains of the Af2 clonal complex of M. bovis have, in general, four or more copies of the insertion sequence IS6110, in contrast to the majority of M. bovis strains isolated from cattle, which are thought to carry only one or a few copies. PMID:21097608

  12. Purification, characterization, and pathogenicity of Moraxella bovis pili.

    Science.gov (United States)

    Ruehl, W W; Marrs, C F; Fernandez, R; Falkow, S; Schoolnik, G K

    1988-09-01

    Pilins composed of the alpha or beta pilins of Moraxella bovis strain Epp63 were purified, subjected to chemical or enzymatic cleavage, and the resulting fragments sequenced by automated Edman degradation. alpha Pilin was found to be a 155-amino-acid polypeptide with a single intramolecular disulfide bridge. The beta pilin amino acid sequence substantiated the previously reported structure derived from the beta pilin gene DNA sequence, and indicated that the alpha and beta pilins of this strain are approximately 70% homologous. DNA hybridization studies of genomic DNA from the alpha- and beta-piliated variants of strain Epp63 indicated that the expression of the two pilin types was governed by an oscillating mechanism of chromosomal rearrangement. The alpha and beta pili were evaluated serologically and found to exhibit approximately 50% shared antigenicity, indicating that regions of conserved and heterologous sequence specify both type-specific and crossreacting epitopes. The pathogenicity of the alpha- and beta-piliated variants was studied by ocular inoculation of calves eyes; beta-piliated organisms were significantly more infectious than alpha-piliated organisms, indicating that beta pili confer, or are associated with, a relative advantage during the first stages of ocular infection. Preliminary analysis of other M. bovis strains suggests that each strain produces two types of pilin, and that this property may be characteristic of the species.

  13. Mycobacterium bovis in Burkina Faso: epidemiologic and genetic links between human and cattle isolates.

    Directory of Open Access Journals (Sweden)

    Adama Sanou

    2014-10-01

    Full Text Available In sub-Saharan Africa, bovine tuberculosis (bTB is a potential hazard for animals and humans health. The goal of this study was to improve our understanding of bTB epidemiology in Burkina Faso and especially Mycobacterium bovis transmission within and between the bovine and human populations.Twenty six M. bovis strains were isolated from 101 cattle carcasses with suspected bTB lesions during routine meat inspections at the Bobo Dioulasso and Ouagadougou slaughterhouses. In addition, 7 M. bovis strains were isolated from 576 patients with pulmonary tuberculosis. Spoligotyping, RDAf1 deletion and MIRU-VNTR typing were used for strains genotyping. The isolation of M. bovis strains was confirmed by spoligotyping and 12 spoligotype signatures were detected. Together, the spoligotyping and MIRU-VNTR data allowed grouping the 33 M. bovis isolates in seven clusters including isolates exclusively from cattle (5 or humans (1 or from both (1. Moreover, these data (genetic analyses and phenetic tree showed that the M. bovis isolates belonged to the African 1 (Af1 clonal complex (81.8% and the putative African 5 (Af5 clonal complex (18.2%, in agreement with the results of RDAf1 deletion typing.This is the first detailed molecular characterization of M. bovis strains from humans and cattle in Burkina Faso. The distribution of the two Af1 and putative Af5 clonal complexes is comparable to what has been reported in neighbouring countries. Furthermore, the strain genetic profiles suggest that M. bovis circulates across the borders and that the Burkina Faso strains originate from different countries, but have a country-specific evolution. The genetic characterization suggests that, currently, M. bovis transmission occurs mainly between cattle, occasionally between cattle and humans and potentially between humans. This study emphasizes the bTB risk in cattle but also in humans and the difficulty to set up proper disease control strategies in Burkina Faso.

  14. Analysis of Streptococcus bovis infections at a monographic oncological centre

    Directory of Open Access Journals (Sweden)

    Lozano TG

    2014-02-01

    Full Text Available The Streptococcus bovis is a Gram-positive, facultative anaerobic, catalase and oxidase negative coccus belonging to the genus Streptococcus. It is part of Streptoccus bovis/ equinus complex and it express the Lancefield antigen D on the surface.This complex has been characterized by molecular biology techniques and specifically by 16S rRNA and sodA gene. Phylogenetic trees based on these techniques are complex and therefore the routine work in laboratories, biochemical techniques are used to identify subspecies if it is necessary.The complex is divided into two subtypes based on biochemical properties: positive mannitol fermentation (biotype I including S. gallolyticus (S. gallolyticus subsp. gallolyticus and S. gallolyticus subsp. macedonicus, mannitol negative and ß-glucuronidase negative (biotype II/ 1, which includes more species (S. infantarius subsp. coli and S. lutetiensis and mannitol negative and ß-glucuronidase positive (biotype II/ 2, with a single species called S. gallolyticus subsp. pasteurianus.Owing to the relationship between colon cancer tumour and Streptococcus bovis, we intend to analyse all isolates in our hospital between the periods of 2010 until March 2013 and analyse tumor epidemiology at our center, in patients infected with this pathogen.Despite the different types of samples and out of the possibility of identification of subspecies, were isolated 14 S. bovis of 14 different patients. The isolates patients were (at the beginning: 4 blood (blood culture, 5 urine, 4 multiple exudates and 1 bronchoalveolar lavage. The proportion of men and women was 8/6. The mean age was 67 years (56±91. Malignant tumor distribution was: 6 prostate cancer, 1 breast cancer, 1 biliary tract, 1 skin, 1, stomach, 1 uterus, 1 vulvar, 1 pyriform sinus and other reproductive organs without specify.The study of antimicrobial in vitro susceptibility was performed by microdilution (MicroScan® WalkAway, Siemens, Sacramento, CA, USA and the

  15. Molecular detection of natural Babesia bovis infection from water buffaloes (Bubalus bubalis) and crossbred cattle

    DEFF Research Database (Denmark)

    Mahmmod, Yasser

    2013-01-01

    Babesia bovis (B. bovis) is a major causative agent of bovine babesiosis, with a considerable worldwide impact. The objective of this study was to evaluate the usefulness of PCR assay and microscopical examination (ME) for detection of B. bovis in naturally infected and apparently healthy water...... buffaloes and crossbred cattle under field circumstances from Sharkia province of Egypt. A total 34 animals (20 crossbred cattle and 14 buffaloes) were clinically and laboratory investigated during the period from March to August 2008. Fifteen animals showed symptoms of bovine babesiosis while 19 animals...

  16. Endocardite infecciosa por Streptococcus bovis em paciente com carcinoma colônico Endocarditis infecciosa por Streptococcus bovis en paciente con carcinoma colónico Infectious endocarditis due to Streptococcus bovis in a patient with colon carcinoma

    Directory of Open Access Journals (Sweden)

    Alexandre Maulaz Barcelos

    2010-09-01

    Full Text Available Descrevemos o caso de uma paciente de 66 anos de idade, com endocardite infecciosa por Streptococcus bovis e adenocarcinoma colônico, que desenvolveu insuficiência aórtica grave aguda. Foi submetida à cirurgia de troca valvar aórtica e posteriormente à ressecção tumoral (hemicolectomia direita. É importante ressaltar a necessidade de complementação do estudo do cólon, mesmo em indivíduos assintomáticos, quando diagnosticamos endocardite infecciosa por S. bovis.Describimos el caso de una paciente de 66 años de edad, con endocarditis infecciosa por streptococcus bovis y adenocarcinoma colónico, que desarrolló insuficiencia aórtica grave aguda. Fue sometida a cirugía de reemplazo valvular aórtico y posteriormente a resección tumoral (hemicolectomía derecha. Es importante destacar la necesidad de complementación del estudio del colon, aun en individuos asintomáticos, cuando diagnosticamos endocarditis infecciosa por S. bovis.We report the case of a 66 year-old female patient with infectious endocarditis due to Streptococcus bovis and adenocarcinoma of the colon that developed acute aortic insufficiency. She was submitted to aortic valve replacement surgery and later to tumor resection (right hemicolectomy. It is important to emphasize the need for complementing the study of the colon, even in asymptomatic individuals, when infectious endocarditis due to S. bovis is diagnosed.

  17. Comparison of screening procedures for Mycobacterium tuberculosis infection among patients with inflammatory diseases

    DEFF Research Database (Denmark)

    Soborg, Bolette; Ruhwald, Morten; Hetland, Merete Lund;

    2009-01-01

    OBJECTIVE: To test if Mycobacterium tuberculosis screening results differ among patients with inflammatory disease depending on whether the QuantiFeron TB-Gold test (QFT) or tuberculin skin test (TST) is used; and to evaluate if a possible difference is influenced by the presence of risk factors...... or immunosuppression. METHODS: The interferon-gamma response to in vitro stimulation of M. tuberculosis-specific antigens was measured with QFT and results were compared with TST. Associations to bacillus Calmette-Guerin (BCG) vaccination, risk factors, and immunosuppression were analyzed for both tests. RESULTS: QFT...... and TST results were available for 294/302 and 241/302 patients, respectively; 234 had results from both tests. Twenty-one (7%) tested positive with QFT and 45 (19%) with TST. A positive QFT was associated with risk factors for M. tuberculosis infection: i.e., birth or upbringing in a TB-endemic area...

  18. Immunological Links to Nonspecific Effects of DTwP and BCG Vaccines on Infant Mortality

    DEFF Research Database (Denmark)

    Claesson, Mogens Helweg

    2011-01-01

    A number of mainly observational studies suggest that many African females below the age of one year die each year from the nonspecific effects of vaccination with diphtheria-tetanus toxoids and killed (whole-cell) Bordetella pertussis (DTwP). In contrast, similar studies suggest that many African...... females and males may have their lives saved each year by the nonspecific immunological benefits of Bacillus Calmette-Guerin (BCG) vaccination. From an immunological point of view, we hypothesise that the adverse effects of DTwP vaccine may occur because of the Th2-polarising effect of the aluminium...... phosphate adjuvant in the vaccine and because intramuscular administration of the vaccine may cause chronic inflammation at the site of injection. However, the Th1-polarising effect of BCG is likely to be beneficial. Sexual dimorphism affecting immune functions and vitamin A supplementation may influence...

  19. Vaccination coverage of patients with inborn errors of metabolism and the attitudes of their parents towards vaccines.

    Science.gov (United States)

    Cerutti, Marta; De Lonlay, Pascale; Menni, Francesca; Parini, Rossella; Principi, Nicola; Esposito, Susanna

    2015-11-27

    To evaluate vaccination coverage of children and adolescents with inborn errors of metabolism (IEMs) and the attitudes of their parents towards vaccination, the vaccination status of 128 patients with IEM and 128 age- and gender-matched healthy controls was established by consulting the official vaccination chart. In children with IEMs, compared with healthy controls, low vaccination rates and/or delays in administration were observed for pneumococcal conjugate, meningococcus C, measles, mumps, rubella, diphtheria-tetanus-pertussis-inactivated polio, Bacillus Calmette-Guerin, and influenza vaccines. Among the parents of IEM patients, vaccine schedule compliance was primarily driven by the doctors at the hospital's reference centres; among the parents of the healthy controls, compliance was driven by the primary care paediatricians. These results show that IEM patients demonstrate sub-optimal vaccination coverage. Further studies of the different vaccines in each IEM disorder and educational programmes aimed at physicians and parents to increase immunization coverage in these patients are urgently needed.

  20. Situation and prophylactic measures of pediatric tuberculosis%儿童结核病与预防

    Institute of Scientific and Technical Information of China (English)

    李蓉

    2015-01-01

    China is a high incidence country of tuberculosis decided by WHO. The infection rates and mortality rates of pediatric tuberculosis are pretty high. Aiming at the features of pediatric tuberculosis, Bacillus calmette-guerin becomes a primary prophylactic measure for the moment. The new research is developed and bound to strike home in new vaccines and chemicals of pediatric tuberculo-sis.%中国是世界卫生组织认定的结核病高发国家之一,儿童结核病的感染率和患病率相对较高,目前主要预防措施是接种卡介苗,新的抗结核疫苗和结核病的化学预防也在研究中,并取得了一定的效果。

  1. BCGiosis as a presenting feature of a child with chronic granulomatous disease

    Directory of Open Access Journals (Sweden)

    Zahra Movahedi

    2011-02-01

    Full Text Available Bacillus Calmette Guerin (BCG vaccine, which is administered to all newborns in some regions, could lead to serious complication ranging from local disease (known as BCGitis to disseminated disease (BCGosis in a group of patients with primary immunodeficiency diseases. We are reporting here a 3.5 year-old girl with a history of prolonged BCGitis, which developed to disseminated disease without any other special features. Immunological studies with nitro-blue tetrazolium test confirmed the diagnosis of chronic granulomatous disease in this patient. Chronic granulomatous disease should be considered in the list of differential diagnosis in all children with BCGosis, even in the absence of any other manifestations related to immunodeficiency.

  2. Isolation and culture of murine macrophages.

    Science.gov (United States)

    Davies, John Q; Gordon, Siamon

    2005-01-01

    The two most convenient sources of primary murine macrophages are the bone marrow and the peritoneal cavity. Resident peritoneal macrophages can readily be harvested from mice and purified by adherence to tissue culture plastic. The injection of Bio-Gel polyacrylamide beads or thioglycollate broth into the peritoneal cavity produces an inflammatory response allowing the purification of large numbers of elicited macrophages. The production of an activated macrophage population can be achieved by using Bacillus-Calmette-Guerin as the inflammatory stimulus. Resident bone marrow macrophages can be isolated following enzymatic separation of cells from bone marrow plugs and enrichment on 30% fetal calf serum containing medium or Ficoll-Hypaque gradients. Bone marrow-derived macrophages can be produced by differentiating nonadherent macrophage precursors with medium containing macrophage colony-stimulating factor.

  3. 卫生部免疫规划专家咨询委员会的作用

    Institute of Scientific and Technical Information of China (English)

    郑景山; 王华庆; 周玉清; 殷大鹏; 梁晓峰

    2010-01-01

    @@ 1 背景我国从1978年开始实施计划免疫,按照统一的免疫程序为适龄儿童提供卡介苗(Bacillus Calmette Guerin,BCG)、口服脊髓灰质炎(脊灰)减毒活疫苗(Oral Poliomyelitis Attenuated Live Vaccine,OPV)、麻疹减毒活疫苗(Measles Attenuated Live Vaccine,MV)和百日咳-白喉-破伤风联合疫苗(Diphtheria,Tetanus,Pertussis Combined Vaccine,DPT)接种,预防结核、脊灰、麻疹、百日咳、白喉、破伤风6种常见传染病.

  4. 一起预防接种实施差错事故引发的思考

    Institute of Scientific and Technical Information of China (English)

    耿晓冬

    2011-01-01

    某患儿,男,2009年4月19日出生,无疫苗接种禁忌证和过敏史,既往已经接种了卡介苗(Bacillus Calmette Guerin,BCG)1剂、乙型病毒性肝炎(乙肝)疫苗(Hepatitis B Vaccine,HepB)2剂、百日咳-白喉-破伤风联合疫苗( Diphtheria,Tetanus,Pertussis Combined Vaccine,DPT)3剂、脊髓灰质炎疫苗(Oral Poliomyelitis Attenuated Live Vaccine,OPV)3剂,家长自述过往接种疫苗未发现异常反应,家族无患病史.

  5. Immunological Links to Nonspecific Effects of DTwP and BCG Vaccines on Infant Mortality

    Directory of Open Access Journals (Sweden)

    Mogens Helweg Claesson

    2011-01-01

    Full Text Available A number of mainly observational studies suggest that many African females below the age of one year die each year from the nonspecific effects of vaccination with diphtheria-tetanus toxoids and killed (whole-cell Bordetella pertussis (DTwP. In contrast, similar studies suggest that many African females and males may have their lives saved each year by the nonspecific immunological benefits of Bacillus Calmette-Guerin (BCG vaccination. From an immunological point of view, we hypothesise that the adverse effects of DTwP vaccine may occur because of the Th2-polarising effect of the aluminium phosphate adjuvant in the vaccine and because intramuscular administration of the vaccine may cause chronic inflammation at the site of injection. However, the Th1-polarising effect of BCG is likely to be beneficial. Sexual dimorphism affecting immune functions and vitamin A supplementation may influence both the deleterious and beneficial nonspecific effects of immunisation.

  6. 禄劝彝族苗族自治县1997~2006年儿童免疫规划5种疫苗接种率调查

    Institute of Scientific and Technical Information of China (English)

    李自雄

    2008-01-01

    @@ 禄劝彝族苗族自治县(禄劝县)1997~2006年每年按照世界卫生组织推荐的按容量比例概率抽样方法,对30个村210名12~23月龄儿童,免疫规划5种疫苗[卡介苗(Bacillus Calmette Guerin,BCG)、口服脊髓灰质炎减毒活疫苗(Oral Poliomylitis Vaccine,Live;OPV)、百白破联合疫苗(Diphtheria,Tetanus and Pertussis Combined Vaccine,DPT)、麻疹减毒活疫苗(Measles Vaccine,Live;MV)、乙型肝炎疫苗(Hepatitus B Vaccine,HepB)]的接种率进行调查,判定标准按卫生部.

  7. The role of vitamin D in malaria.

    Science.gov (United States)

    Lương, Khanh Vinh Quốc; Nguyễn, Lan Thi Hoàng

    2015-01-15

    An abnormal calcium-parathyroid hormone (PTH)-vitamin D axis has been reported in patients with malaria infection. A role for vitamin D in malaria has been suggested by many studies. Genetic studies have identified numerous factors that link vitamin D to malaria, including human leukocyte antigen genes, toll-like receptors, heme oxygenase-1, angiopoietin-2, cytotoxic T lymphocyte antigen-4, nucleotide-binding oligomerization domain-like receptors, and Bcl-2. Vitamin D has also been implicated in malaria via its effects on the Bacillus Calmette-Guerin (BCG) vaccine, matrix metalloproteinases, mitogen-activated protein kinase pathways, prostaglandins, reactive oxidative species, and nitric oxide synthase. Vitamin D may be important in malaria; therefore, additional research on its role in malaria is needed.

  8. Sex-based differences in immune function and responses to vaccination

    Science.gov (United States)

    Klein, Sabra L.; Marriott, Ian; Fish, Eleanor N.

    2015-01-01

    Females typically develop higher antibody responses and experience more adverse reactions following vaccination than males. These differences are observed in response to diverse vaccines, including the bacillus Calmette-Guerin vaccine, the measles, mumps and rubella vaccine, the yellow fever virus vaccine and influenza vaccines. Sex differences in the responses to vaccines are observed across diverse age groups, ranging from infants to aged individuals. Biological as well as behavioral differences between the sexes are likely to contribute to differences in the outcome of vaccination between the sexes. Immunological, hormonal, genetic and microbiota differences between males and females may also affect the outcome of vaccination. Identifying ways to reduce adverse reactions in females and increase immune responses in males will be necessary to adequately protect both sexes against infectious diseases. PMID:25573105

  9. 1例卡介苗接种事故的处理报告

    Institute of Scientific and Technical Information of China (English)

    张雪梅; 朱俞学; 邢茹琳

    2009-01-01

    @@ 患儿,女,2002年1月10日出生,生长发育良好.2008年8月18日14:00到当地预防接种点,准备接种白喉-破伤风联合疫苗(Diphthiria and Tetanus Combined Vaccine,DT),但接种医生误将另一支稀释好待用的剩余量0.4ml的卡介苗(Bacillus Calmette Guerin,BCG;成都生物制品研究所生产,批号200702a023-1,有效期至2009年2月25日),注入患儿左上臂三角肌内.

  10. Isolation of Streptococcus bovis in a Patient with Undiagnosed Colon Cancer. A Case Report

    Directory of Open Access Journals (Sweden)

    Ledys Pérez Morales

    2014-02-01

    Full Text Available One of the bacterial agents commonly associated with colorectal cancer is Streptococcus bovis. Twenty five to 80 % of patients with Streptococcus bovis bacteremia develop colorectal tumors and the incidence of colonic neoplasia associated with S. bovis endocarditis has been shown to be 18 to 62 %. Hence, it was decided to present the case of a 57-year-old patient from an urban area with a 6-year history of diabetes mellitus type I and alcoholism, who was admitted to the hospital in Cienfuegos because of significant gastrointestinal bleeding and fever (39-40 o C. Streptococcus bovis was isolated from the blood culture. The patient died 72 hours after his admission. Autopsy results showed a moderately differentiated adenocarcinoma of the right colon.

  11. Multinucleated giant cell cytokine expression in pulmonary granulomas of cattle experimentally infected with Mycobacterium bovis

    Science.gov (United States)

    Pathogenic mycobacteria of the Mycobacterium tuberculosis complex such as Mycobacterium bovis, induce a characteristic lesion known as a granulomas. Granulomas represent a specific host response to chronic antigenic stimuli, such as foreign bodies, certain bacterial components, or persistent pathoge...

  12. Current knowledge and pending challenges in zoonosis caused by Mycobacterium bovis: a review.

    Science.gov (United States)

    Pérez-Lago, Laura; Navarro, Yurena; García-de-Viedma, Darío

    2014-10-01

    Mycobacterium bovis is both the causative agent of bovine tuberculosis (TB) and a zoonotic pathogen. In humans, considerably fewer cases of TB are caused by M. bovis than M. tuberculosis; nevertheless, diagnostic limitations mean that currently available data on prevalence grossly underestimate the true dimension of the problem. The routes of transmission from animals to humans are well known and include direct exposure to infected animals or consumption of contaminated animal products. Application of fingerprinting tools facilitates analysis of the molecular epidemiology of M. bovis in animal-to-human and human-to-human transmission. Apart from cattle and M. bovis, other animal species and members within the M. tuberculosis complex can contribute to the zoonosis. Improvements in diagnostic techniques, application of more advanced discriminatory genotyping tools, and collaboration between veterinary and human health care researchers are key to our understanding of this zoonosis.

  13. Effects of Moraxella bovis vaccination schedules on experimentally induced infectious bovine keratoconjunctivitis.

    Science.gov (United States)

    Webber, J J; Selby, L A

    1981-07-01

    An oil-adjuvant Moraxella bovis bacterin was administered to weanling calves, using different vaccination schedules. Calves were given a booster vaccination after 3 weeks and were challenge exposed 2 weeks later with virulent M bovis recovered from calves with clinical infectious bovine keratoconjunctivitis (IBK). The effects of different routes of vaccination and homologous and heterologous challenge exposure on the incidence, severity, and duration of induced IBK was evaluated. All calves given a placebo developed clinical IBK. Calves vaccinated subcutaneously in the neck had the shortest duration of M bovis infection, the lowest incidence and the shortest duration of acute IBK, and the lowest disease severity score, compared with effects in calves given a placebo or vaccinated subconjunctivally. Calves challenge exposed with the homologous strain of M bovis had more infected eyes, more eyes with acute IBK, longer duration of infection, and a higher severity and duration disease score.

  14. Experimental infectious bovine keratoconjunctivitis: efficacy of a vaccine prepared from nonhemolytic strains of Moraxella bovis.

    Science.gov (United States)

    Pugh, G W; McDonald, T J; Kopecky, K E

    1982-06-01

    Calves vaccinated with a bacterin prepared from 4 nonhemolytic strains of Moraxella bovis were challenge exposed with a virulent hemolytic strain of M bovis. Ten calves were given 2 vaccinal inoculations 21 days apart. Twenty-one days after the last vaccinal inoculation, the eyes of the 10 vaccinated and the eyes of the 12 non-vaccinated calves were exposed to a hemolytic heterologous strain of M bovis. The percentage of eyes with infectious bovine keratoconjunctivitis was lower, the lesions were less severe, and the duration of infection was shorter in vaccinated calves than in nonvaccinated calves. Seemingly, the resistance induced by nonhemolytic strains of M bovis was at least comparable with the induced by hemolytic virulent strains of that organism. The nature of the immune response in vaccinated calves is discussed.

  15. Amplification of a 500-Base-Pair Fragment from Cultured Isolates of Mycobacterium bovis

    Science.gov (United States)

    Rodríguez, Juan Germán; Fissanoti, Juan Carlos; Del Portillo, Patricia; Patarroyo, Manuel Elkin; Romano, María Isabel; Cataldi, Angel

    1999-01-01

    The presence of a 500-bp fragment which amplifies a region from the genome of Mycobacterium bovis (J. G. Rodriguez, G. A. Meija, P. Del Portillo, M. E. Patarroyo, and L. A. Murillo, Microbiology 141:2131–2138, 1995) was evaluated by carrying out PCR on 121 M. bovis isolates. The M. bovis strains, previously characterized by culture and biochemical tests, were isolated from cattle in different regions of Argentina, Mexico, and Colombia. Four additional strains isolated from sea lions that belong to the M. tuberculosis complex were also included in the study. All of the isolates tested were PCR positive, rendering the expected 500-bp band and giving a correlation of 100% with previous microbiological characterization. Southern blot analysis revealed a common band of 1,800 bp and a polymorphic high-molecular-mass hybridization pattern. The results show that this assay may be useful for diagnosis and identification of M. bovis in cattle. PMID:10364607

  16. High Mobility Group Box 1 Protein Induction by Mycobacterium Bovis BCG

    Directory of Open Access Journals (Sweden)

    Péter Hofner

    2007-01-01

    Conclusion: Our pilot experiments draw attention to the HMGB1 inducing ability of Mycobacterium bovis. Assesment of the pathophysiological role of this late cytokine in mycobacterial infections demands further in vitro and in vivo examinations.

  17. Molecular Typing of Mycobacterium bovis from Cattle Reared in Midwest Brazil

    Science.gov (United States)

    Carvalho, Ricardo César Tavares; Vasconcellos, Sidra Ezidio Gonçalves; Issa, Marina de Azevedo; Soares Filho, Paulo Martins; Mota, Pedro Moacyr Pinto Coelho; de Araújo, Flábio Ribeiro; Carvalho, Ana Carolina da Silva; Gomes, Harrison Magdinier; Suffys, Philip Noel; Paschoalin, Vânia Margaret Flosi

    2016-01-01

    Mycobacterium bovis is the causative agent of bovine tuberculosis (BTB), the pathogen responsible for serious economic impact on the livestock sector. In order to obtain data on isolated M. bovis strains and assist in the control and eradication program for BTB, a cross sectional descriptive molecular epidemiology study in the Brazilian Midwest was conducted. Through spoligotyping and 24-loci MIRU-VNTR methods, 37 clinical isolates of M. bovis circulating in the region were analyzed, 10 isolated from the state of Mato Grosso, 12 from the state of Mato Grosso do Sul and 15 from the state of Goiás. The spoligotyping analysis identified 10 distinct M. bovis profiles (SB0121 n = 14, SB0295 n = 6, SB0140 n = 6, SB0881 n = 3, SB1144 n = 2, SB1145 n = 2, SB0134 n = 1, SB1050 n = 1, SB1055 n = 1, SB1136 n = 1) grouped in six clusters and four orphan patterns. The MIRU-VNTR 24-loci grouped the same isolates in six clusters and 22 unique orphan patterns, showing higher discriminatory power than spoligotyping. When associating the results of both techniques, the isolates were grouped in five clusters and 24 unique M. bovis profiles. Among the 24-loci MIRU-VNTR evaluated, two, ETR-A and QUB 11b loci, showed high discriminatory ability (h = ≥ 0.50), while MIRU 16, MIRU 27, ETR-B, ETR-C, Mtub21 and QUB 26 loci showed moderate ability (h = 0.33 or h = 0.49) and were the most effective in evaluating the genotypic similarities among the clinical M. bovis isolate samples. Herein, the 29 patterns found amongst the 37 isolates of M. bovis circulating in the Brazilian Midwest can be due to the animal movement between regions, municipalities and farms, thus causing the spread of various M. bovis strains in herds from Midwest Brazil. PMID:27631383

  18. Protection against tuberculosis in Eurasian wild boar vaccinated with heat-inactivated Mycobacterium bovis.

    Directory of Open Access Journals (Sweden)

    Joseba M Garrido

    Full Text Available Tuberculosis (TB caused by Mycobacterium bovis and closely related members of the Mycobacterium tuberculosis complex continues to affect humans and animals worldwide and its control requires vaccination of wildlife reservoir species such as Eurasian wild boar (Sus scrofa. Vaccination efforts for TB control in wildlife have been based primarily on oral live BCG formulations. However, this is the first report of the use of oral inactivated vaccines for controlling TB in wildlife. In this study, four groups of 5 wild boar each were vaccinated with inactivated M. bovis by the oral and intramuscular routes, vaccinated with oral BCG or left unvaccinated as controls. All groups were later challenged with a field strain of M. bovis. The results of the IFN-gamma response, serum antibody levels, M. bovis culture, TB lesion scores, and the expression of C3 and MUT genes were compared between these four groups. The results suggested that vaccination with heat-inactivated M. bovis or BCG protect wild boar from TB. These results also encouraged testing combinations of BCG and inactivated M. bovis to vaccinate wild boar against TB. Vaccine formulations using heat-inactivated M. bovis for TB control in wildlife would have the advantage of being environmentally safe and more stable under field conditions when compared to live BCG vaccines. The antibody response and MUT expression levels can help differentiating between vaccinated and infected wild boar and as correlates of protective response in vaccinated animals. These results suggest that vaccine studies in free-living wild boar are now possible to reveal the full potential of protecting against TB using oral M. bovis inactivated and BCG vaccines.

  19. Protection against tuberculosis in Eurasian wild boar vaccinated with heat-inactivated Mycobacterium bovis.

    Science.gov (United States)

    Garrido, Joseba M; Sevilla, Iker A; Beltrán-Beck, Beatriz; Minguijón, Esmeralda; Ballesteros, Cristina; Galindo, Ruth C; Boadella, Mariana; Lyashchenko, Konstantin P; Romero, Beatriz; Geijo, Maria Victoria; Ruiz-Fons, Francisco; Aranaz, Alicia; Juste, Ramón A; Vicente, Joaquín; de la Fuente, José; Gortázar, Christian

    2011-01-01

    Tuberculosis (TB) caused by Mycobacterium bovis and closely related members of the Mycobacterium tuberculosis complex continues to affect humans and animals worldwide and its control requires vaccination of wildlife reservoir species such as Eurasian wild boar (Sus scrofa). Vaccination efforts for TB control in wildlife have been based primarily on oral live BCG formulations. However, this is the first report of the use of oral inactivated vaccines for controlling TB in wildlife. In this study, four groups of 5 wild boar each were vaccinated with inactivated M. bovis by the oral and intramuscular routes, vaccinated with oral BCG or left unvaccinated as controls. All groups were later challenged with a field strain of M. bovis. The results of the IFN-gamma response, serum antibody levels, M. bovis culture, TB lesion scores, and the expression of C3 and MUT genes were compared between these four groups. The results suggested that vaccination with heat-inactivated M. bovis or BCG protect wild boar from TB. These results also encouraged testing combinations of BCG and inactivated M. bovis to vaccinate wild boar against TB. Vaccine formulations using heat-inactivated M. bovis for TB control in wildlife would have the advantage of being environmentally safe and more stable under field conditions when compared to live BCG vaccines. The antibody response and MUT expression levels can help differentiating between vaccinated and infected wild boar and as correlates of protective response in vaccinated animals. These results suggest that vaccine studies in free-living wild boar are now possible to reveal the full potential of protecting against TB using oral M. bovis inactivated and BCG vaccines.

  20. [Moraxella bovis biomass production in a bench-top fermentor].

    Science.gov (United States)

    González, R D; Oberti, E R

    1994-01-01

    A Moraxella bovis strain was isolated from a kerato-conjunctivities lesion of a calf in Villa Valeria (Córdoba); it was used to establish improved cultural conditions, such as nature and concentration of carbon and nitrogen sources, and pH control in shaken flasks. The selected conditions were assayed for biomass production in a bench-top fermentor. The strain is used by the pharmaceutical industry to produce vaccines and adjuvants. In the initial condition (48 h culture on blood agar) 0.019 g biomass/l.h-1 was obtained. With the use of liquid defined medium with pH control, productivity was increased to 0.153g/l.h-1, with optimum harvest time of 32 h.

  1. Antibiotic sensitivity of an Argentine strain collection of Moraxella bovis.

    Science.gov (United States)

    Zielinski, G; Piscitelli, H; Perez-Monti, H; Stobbs, L A

    2000-01-01

    The antimicrobial susceptibility of 88 isolates of Moraxella bovis of Argentine origin was evaluated for 12 antimicrobials by broth microdilution procedures. The isolates had a minimum inhibitory concentration (MIC90) of or = 32 microg/mL to lincomycin. Modal MIC values for these antimicrobials were as follows: enrofloxacin, 0.03 microg/mL; ceftiofur, 0.06 pg/mL; ampicillin, 0.25 microg/mL; florfenicol, gentamicin, erythromycin, and oxytetracycline, 0.5 microg/mL; tilmicosin, 1.0 microg/mL; tylosin and spectinomycin, 4.0 microg/mL; lincomycin and erythromycin, 16 microg/mL; and trimethoprim/ sulfamethoxazole, < or = 0.25/4.75 microg/mL. These data show that all antimicrobials except lincomycin have MICs suggestive of sensitivity in vitro, though confirmation of clinical efficacy can only be properly assessed based on pharmacologic and/or clinical data to support the MIC values.

  2. Safety evaluation in mice of the childhood immunization vaccines from two south-eastern states of Nigeria

    Institute of Scientific and Technical Information of China (English)

    Oli; Angus; Nnamdi; Agu; Remigus; Uchenna; Oli; Ugochukwu; Chinedum; Nwoye; Charles; Ugochukwu; Ejiofor; Obiora; Shedrack; Esimone; Charles; Okechukwu

    2015-01-01

    Objective:To check the effects of the vaccines on the hematopoietic system and weight of mice after immunization.Methods:The study was done with the Expanded Programme on Immunization vaccines donated by the Ministries of Health of Abia and Imo States of Nigeria.The vaccines were collected from the cold-chain stores and transported in vaccine carriers to the cold-chain facility in Nnamdi Azikiwe University Teaching Hospital within 3 hours of collection.They were used to immunize a total of 160 mice.The Ethics Committee of Nnamdi Azikiwe University Teaching Hospital,Nnewi of Anambra State,Nigeria approved the protocol.Results:Mice body weight changes test showed that the mice all had increased body weight at Days 3 and 7 post-immunization and none died during the 7 d post-immunization observation.The percentage weight gains of the mice compared with the control were 69%.70%,64%.63%,65%and 68%for oral polio vaccine,diphtheria-pertussis-tetanus.bacillus CalmetteGuerin,measles,yellow fever and hepatitis B vaccines respectively collected from Imo State.The mice immunized with oral polio vaccine,pentavalent.bacillus Calmette-Guerin.measles,yellow fever and hepatitis B vaccines collected from Abia State had 123%.114%,121%.116%,142%and 119%weight gain respectively compared with the control.Leukocytosis promoting toxicity test showed that none of the vaccines was able to induce proliferation of leukocytes up to ten folds.Leukopenic toxicity test showed that all the vaccines had an leukopenic toxicity test value higher than 80%of the control(physiological saline).Conclusions:The vaccine samples tested were safe and did not affect the hematopoietic system adversely.The storage conditions of the vaccines in the States’ cold-chain stores had not compromised the safety of the vaccines.

  3. Prevalence of Mycoplasma bovis in Respiratory Tract of Cattle Slaughtered in Balochistan, Pakistan

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    Zafar Ahmad

    2014-01-01

    Full Text Available Cattle lungs (n=1200 obtained from abattoir of 10 districts of Balochistan were processed for isolation and identification of Mycoplasma species. A total of 156 isolates produced typical fried egg colonies on Modified Hayflick’s agar medium and 87.8% were preliminarily identified as Mycoplasma species, 12.2% species were Acholeplasmas. All the digitonin sensitive isolates were further subjected to different biochemical and PCR tests for further identification. Overall prevalence of M. bovis lungs samples obtained from slaughter house samples was 9%. Among the Mycoplasma isolates; 108 M. bovis, 29 Mycoplasma mycoides subsp. capri (Mmc and 16 M. arginini were identified through the biochemical tests. M. bovis and Mycoplasma mycoides subcluster members were further validated through PCR and RFLP. Mycoplasma mycoides subspecies mycoides small colony type (Mmm SC was not isolated from any of the lung samples. Among the Mycoplasma bovis species isolated, the highest number was observed from Quetta district (16% followed by Pishin (15%, Zhob (11 % and Kalat (10%. Conversely the lowest number of M. bovis isolates was found in Bolan (2% district followed by Jaffarabad (3%, 4%, each from Khuzdar, Mustung, Killasaifullah and 7% in Sibi district. Statistical analysis using chi square test, showed a significance difference (χ²=33.38 in the recovery of Mycoplasma bovis from the lungs of cattle slaughtered in 10 districts of Balochistan.

  4. Investigation of the effect of Mycobacterium bovis infection on bovine neutrophils functions.

    Science.gov (United States)

    Wang, Jin; Zhou, Xiangmei; Pan, Bo; Yang, Lifeng; Yin, Xiaomin; Xu, Binrui; Zhao, Deming

    2013-11-01

    Bovine tuberculosis is a disease in cattle caused by infection with Mycobacterium bovis. The disease has posed significant economic losses and remains a public health hazard worldwide. Interactions between M. bovis and bovine macrophages have been extensively characterized in various studies, while similar analyses in neutrophils, which are one of the other types of white blood cells in mammals, were often overlooked. Neutrophils provide defense against all microbes and can present a diverse collection of antimicrobial molecules, which play an important role in the control of tuberculosis progression. Much of the available data about the involvement of neutrophils in the killing M. bovis is controversial. In this study, we assessed the effect of in vitro infection with M. bovis on some parameters of neutrophils functions including phenotypic changes, apoptosis rate and inflammatory cytokines production. Our results demonstrated that phagocytosis of M. bovis activated and enhanced bovine neutrophils functions as well as initialed their defense mechanism, but failed to eliminate the mycobacteria. Moreover, autophagy might get involved in the defense infection process functioning as a protective mechanism, and inducible-autophagy by lipopolysaccharides stimulation and starvation treatment could efficiently reverse the inability of neutrophils for killing M. bovis, suggesting a potential target for anti-mycobacterial drug-therapy.

  5. Pathogenesis of corneal lesions caused by Moraxella bovis in gnotobiotic calves.

    Science.gov (United States)

    Rogers, D G; Cheville, N F; Pugh, G W

    1987-07-01

    Moraxella bovis was instilled into the conjunctival sac of gnotobiotic calves and corneas were sampled serially after infection. Lesions developed in seven of eight infected calves, but were absent in a noninfected control calf. Histologically, M. bovis was first seen in foci of swollen epithelium and within basal epithelial cells adjacent to ulcers. Corneal ulcers were severe in later stages of infection; fibrin deposits, neutrophils, and bacteria were present in the stromas. Examination of early lesions by scanning electron microscopy showed M. bovis in pits on the surfaces of dark epithelial cells, enmeshed in degenerate epithelial cells and within erosions and an ulcer; in later samples, bacteria were rare. Ultrastructurally, M. bovis was seen in surface pits in superficial epithelial cell processes and within swollen epithelial cells. In stroma, M. bovis was frequently seen among collagen fibrils, within neutrophil phagosomes, and associated with cellular debris. This study demonstrates that a virulent strain of M. bovis can invade bovine corneal epithelial cells and can cause keratitis in the absence of injurious ultraviolet irradiation or other known predisposing environmental factors.

  6. Characterization of hemolysin of Moraxella bovis using a hemolysis-neutralizing monoclonal antibody.

    Science.gov (United States)

    Billson, F M; Harbour, C; Michalski, W P; Tennent, J M; Egerton, J R; Hodgson, J L

    2000-06-01

    A concentrated bacterial culture supernatant from the hemolytic Moraxella bovis strain UQV 148NF was used to immunize mice and generate monoclonal antibodies (MAbs). One, MAb G3/D7, neutralized the hemolytic activity of M. bovis and recognized a 94-kDa protein by Western blot analysis in hemolytic M. bovis strains representing each of the different fimbrial serogroups. Exposure of corneal epithelial cells to M. bovis concentrated culture supernatants demonstrated a role for an exotoxin in the pathogenesis of infectious bovine keratoconjunctivitis, while neutralization of hemolytic and cytotoxic activities by MAb G3/D7 implies that these activities are related or have common epitopes. The action of M. bovis hemolysin was further characterized in sheep erythrocyte preparations with a binding step and Ca(2+) required for lysis to proceed, similar to the RTX family of bacterial exotoxins. Neutralization of lytic activity in vitro is evidence for the presence of M. bovis antigens, which may be capable of protecting cattle from the development of infectious bovine keratoconjunctivitis.

  7. Draft Genome Sequences of Mycobacterium bovis BZ 31150 and Mycobacterium bovis B2 7505, Pathogenic Bacteria Isolated from Archived Captive Animal Bronchial Washes and Human Sputum Samples in Uganda.

    Science.gov (United States)

    Wanzala, Sylvia I; Nakavuma, Jesca; Travis, Dominic A; Kia, Praiscillia; Ogwang, Sam; Sreevatsan, Srinand

    2015-10-08

    Bovine tuberculosis (BTB), a zoonotic infection of cattle caused by Mycobacterium bovis, results in losses of $3 billion to the global agricultural industry and represents the fourth most important livestock disease worldwide. M. bovis as a source of human infection is likely underreported due to the culture medium conditions used to isolate the organism from sputum or other sample sources. We report here the draft genome sequences of M. bovis BZ 31150, isolated from a bronchial washing from a captive chimpanzee, and M. bovis B2 7505, isolated from a human sputum sample in Uganda.

  8. M2-Polarized Macrophages Compose Lupus Vulgaris Arising from a Bacillus Calmette-Guérin Vaccination Site.

    Science.gov (United States)

    Sato, Yota; Fujimura, Taku; Furudate, Sadanori; Kakizaki, Aya; Iizawa, Osamu; Aiba, Setsuya

    2016-01-01

    Since bacillus Calmette-Guérin (BCG) is an attenuated strain of Mycobacterium bovis, lupus vulgaris (LV) is reported as one of the rare complications after BCG vaccination, correlating with immunosuppression in the lesional skin. In this report, we describe a case of LV arising from the BCG vaccination site 22 years after vaccination. Interestingly, in the present case, granuloma cells were composed of M2-polarized macrophages. Our case might explain the contribution of M2-polarized macrophages to the biology of LV arising from a BCG vaccination site.

  9. Immunogenicity of a Moraxella bovis bacterin containing attachment and cornea-degrading enzyme antigens.

    Science.gov (United States)

    Gerber, J D; Selzer, N L; Sharpee, R L; Beckenhauer, W H

    1988-02-01

    An adjuvanted Moraxella bovis bacterin containing attachment antigens and cornea-degrading enzyme antigens protected cattle from infectious bovine keratoconjunctivitis (IBK) when experimentally challenged with homologous and heterologous challenge cultures of M. bovis. This bacterin also protected cattle against field exposure to M. bovis. Transmission electron microscopy and fluorescein labeled anti-M. bovis pili antiserum showed pili on the M. bovis bacterin strain. Scanning electron microscopy demonstrated a fibrillar glycocalyx. The bacterin strain of M. bovis, but not all strains of M. bovis, destroyed bovine corneal cell monolayers in vitro. Bovine corneal cells began to separate from each other within 5 min after M. bovis organisms were added and adhered to the cell monolayers. Moraxella bovis organisms remained attached to the disintegrating cells as the cell membrane separated and was digested. Vaccination stimulated bacterial agglutination antibodies. However, protection against experimental challenge was more closely related to the cornea-degrading enzyme content of the experimental bacterins. Twenty-two of 29 cattle (76%) vaccinated with bacterins containing a relative enzyme activity (REA) greater than 0.4 were protected in a rigorous challenge of immunity test. Only 1 of 21 non-vaccinated calves (5%) was free of IBK. Ninety-two percent (24/26) of calves vaccinated with a bacterin containing a REA greater than 0.29 remained free of IBK following field exposure, whereas 47% (8/17) non-vaccinated calves developed IBK. Only 8 of 12 calves (67%) vaccinated with a bacterin containing a REA of 0.09 remained free of IBK. In a larger field efficacy test consisting of 32 herds in six states, the incidence of IBK in individual herds ranged from 0% to 55%. The overall rate of infection was 11.2%. Vaccination of calves with an M. bovis bacterin that contained a REA of 0.63 reduced the incidence of IBK from 11.2% (217/1931) in the non-vaccinated controls to 4

  10. Transfected Babesia bovis Expressing a Tick GST as a Live Vector Vaccine

    Science.gov (United States)

    Oldiges, Daiane P.; Laughery, Jacob M.; Tagliari, Nelson Junior; Leite Filho, Ronaldo Viana; Davis, William C.; da Silva Vaz, Itabajara; Termignoni, Carlos; Knowles, Donald P.; Suarez, Carlos E.

    2016-01-01

    The Rhipicephalus microplus tick is a notorious blood-feeding ectoparasite of livestock, especially cattle, responsible for massive losses in animal production. It is the main vector for transmission of pathogenic bacteria and parasites, including Babesia bovis, an intraerythrocytic apicomplexan protozoan parasite responsible for bovine Babesiosis. This study describes the development and testing of a live B. bovis vaccine expressing the protective tick antigen glutathione-S-transferase from Haemaphysalis longicornis (HlGST). The B. bovis S74-T3B parasites were electroporated with a plasmid containing the bidirectional Ef-1α (elongation factor 1 alpha) promoter of B. bovis controlling expression of two independent genes, the selectable marker GFP-BSD (green fluorescent protein–blasticidin deaminase), and HlGST fused to the MSA-1 (merozoite surface antigen 1) signal peptide from B. bovis. Electroporation followed by blasticidin selection resulted in the emergence of a mixed B. bovis transfected line (termed HlGST) in in vitro cultures, containing parasites with distinct patterns of insertion of both exogenous genes, either in or outside the Ef-1α locus. A B. bovis clonal line termed HlGST-Cln expressing intracellular GFP and HlGST in the surface of merozoites was then derived from the mixed parasite line HlGST using a fluorescent activated cell sorter. Two independent calf immunization trials were performed via intravenous inoculation of the HlGST-Cln and a previously described control consisting of an irrelevant transfected clonal line of B. bovis designated GFP-Cln. The control GFP-Cln line contains a copy of the GFP-BSD gene inserted into the Ef-1α locus of B. bovis in an identical fashion as the HIGST-Cln parasites. All animals inoculated with the HlGST-Cln and GFP-Cln transfected parasites developed mild babesiosis. Tick egg fertility and fully engorged female tick weight was reduced significantly in R. microplus feeding on HlGST-Cln-immunized calves

  11. Isolation and molecular characterization of Mycobacterium bovis from Kafue lechwe (Kobus leche kafuensis) from Zambia.

    Science.gov (United States)

    Malama, Sydney; Johansen, Tone Bjordal; Muma, John Bwalya; Mwanza, Sydney; Djønne, Berit; Godfroid, Jacques

    2014-01-01

    Bovine tuberculosis (BTB) is a chronic bacterial disease caused by Mycobacterium bovis. Infections due to M. bovis, which serves as a stable reservoir, can pose serious challenge to control and eradicate in both wildlife and livestock at the interface. This study aimed at isolating and characterizing M. bovis from Kafue lechwe (Kobus leche kafuensis) and black lechwe (Kobus leche smithemani) at the animal/human interface in Zambia. The samples with lesions compatible with BTB collected during the hunting seasons of 2009 and 2010 were cultured for isolation of mycobacteria using Stonebrink with pyruvate (BD Diagnostics, MD, USA) and Middlebrook 7H10 (BD Diagnostics) slants. Isolated mycobacteria were identified using IS6110 polymerase chain reaction and deletion analysis. Molecular characterization of the isolates was performed using spoligotyping and mycobacteria interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) with nine loci. Data was analyzed using BioNumerics software 6.1. Out of the 39 samples, acid fast bacilli were detected in 27 (69.2 %) based on smear microscopy. Seven isolates were found to belong to Mycobacterium tuberculosis complex, and all were identified as M. bovis based on deletion analysis. All seven isolates were identical on spoligotyping as belonging to the SB0120 (SIT 482). MIRU-VNTR differentiated the isolates into five different patterns. This study has confirmed that M. bovis circulates in the Kafue lechwe, and non-tuberculous mycobacteria were detected in the black lechwe in Zambia which represents a wildlife reservoir, with a potential to spillover to cattle and humans. Isolates of M. bovis from lechwe antelopes are much conserved as only one spoligotype was detected. The study has shown that three loci differentiated fairly well. This option is cheap and less laborious, and hence a better option in resource-strained country like Zambia. The study further showed that some of the loci recommended by the European

  12. Gliding motility of Babesia bovis merozoites visualized by time-lapse video microscopy.

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    Masahito Asada

    Full Text Available BACKGROUND: Babesia bovis is an apicomplexan intraerythrocytic protozoan parasite that induces babesiosis in cattle after transmission by ticks. During specific stages of the apicomplexan parasite lifecycle, such as the sporozoites of Plasmodium falciparum and tachyzoites of Toxoplasma gondii, host cells are targeted for invasion using a unique, active process termed "gliding motility". However, it is not thoroughly understood how the merozoites of B. bovis target and invade host red blood cells (RBCs, and gliding motility has so far not been observed in the parasite. METHODOLOGY/PRINCIPAL FINDINGS: Gliding motility of B. bovis merozoites was revealed by time-lapse video microscopy. The recorded images revealed that the process included egress of the merozoites from the infected RBC, gliding motility, and subsequent invasion into new RBCs. The gliding motility of B. bovis merozoites was similar to the helical gliding of Toxoplasma tachyzoites. The trails left by the merozoites were detected by indirect immunofluorescence assay using antiserum against B. bovis merozoite surface antigen 1. Inhibition of gliding motility by actin filament polymerization or depolymerization indicated that the gliding motility was driven by actomyosin dependent process. In addition, we revealed the timing of breakdown of the parasitophorous vacuole. Time-lapse image analysis of membrane-stained bovine RBCs showed formation and breakdown of the parasitophorous vacuole within ten minutes of invasion. CONCLUSIONS/SIGNIFICANCE: This is the first report of the gliding motility of B. bovis. Since merozoites of Plasmodium parasites do not glide on a substrate, the gliding motility of B. bovis merozoites is a notable finding.

  13. Development and host compatibility of plasmids for two important ruminant pathogens, Mycoplasma bovis and Mycoplasma agalactiae.

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    Shukriti Sharma

    Full Text Available Mycoplasma bovis is a cause of pneumonia, mastitis, arthritis and otitis media in cattle throughout the world. However, despite its clinical significance, there is a paucity of tools to genetically manipulate it, impeding our capacity to further explore the molecular basis of its virulence. To address this limitation, we developed a series of homologous and heterologous replicable plasmids from M. bovis and M. agalactiae. The shortest replicable oriC plasmid based on the region downstream of dnaA in M. bovis was 247 bp and contained two DnaA boxes, while oriC plasmids based on the region downstream of dnaA in M. agalactiae strains 5632 and PG2 were 219 bp and 217 bp in length, respectively, and contained only a single DnaA box. The efficiency of transformation in M. bovis and M. agalactiae was inversely correlated with the size of the oriC region in the construct, and, in general, homologous oriC plasmids had a higher transformation efficiency than heterologous oriC plasmids. The larger pWholeoriC45 and pMM21-7 plasmids integrated into the genomic oriC region of M. bovis, while the smaller oriC plasmids remained extrachromosomal for up to 20 serial passages in selective media. Although specific gene disruptions were not be achieved in M. bovis in this study, the oriC plasmids developed here could still be useful as tools in complementation studies and for expression of exogenous genes in both M. bovis and M. agalactiae.

  14. Sequence conservation of the 12D3 gene in Mexican isolates of Babesia bovis.

    Science.gov (United States)

    Perez, J; Javier Perez, J; Vargas, P; Antonio Alvarez, J; Rojas, C; Figueroa, J V

    2010-04-01

    The 12D3 antigen present in Babesia bovis has been evaluated as a recombinant vaccine candidate and the 12d3 coding sequence has been reported for an Australian and an USA (Texas) isolate of B. bovis. However, no approach has been conducted to perform analysis of 12d3 sequence conservation on a larger number of B. bovis isolates. This could provide important information to determine whether a recombinant vaccine containing this antigen could be widely used. This study reports the cloning and sequencing analysis of the 12d3 coding region in 20 different B. bovis isolates collected from various geographical regions in the tropics and subtropics of Mexico. Comparative analysis of the consensus nucleotide sequences obtained for each isolate revealed a high degree of conservation (94-99% sequence identity) among the 12d3 alleles present in the Mexican isolates when compared with the 12d3 ORF sequences from the Texan (T2Bo) B. bovis isolate. Similarly, BLASTX sequence homology search showed a high percent identity (93-99%) of the deduced amino acid 12D3 sequence as compared with the T2Bo isolate sequence. The high level of sequence conservation in 12d3 among the 20 B. bovis isolates collected from geographically distant locations in Mexico suggests that there exists a minimal bovine-host immunological pressure which could be translated into antigenic diversity or variation, and most probably this is reflected in the non-inmunodominant characteristic of the 12D3 antigen as it has been previously described in the literature. 12D3 antigen can be considered as a viable candidate for inclusion in a recombinant vaccine for cattle babesiosis caused by B. bovis in Mexico.

  15. Comparative characterization of the leukocidic and hemolytic activity of Moraxella bovis.

    Science.gov (United States)

    Hoien-Dalen, P S; Rosenbusch, R F; Roth, J A

    1990-02-01

    The cytotoxic effect of Moraxella bovis 118F on bovine neutrophils was evaluated and characterized by use of a 51Cr release assay. Neutrophils harvested from healthy adult cattle were labeled with 51Cr. The leukocidic activity produced by M bovis 118F, a hemolytic strain of M bovis, was heat-labile. A live culture of strain 118F, at a ratio of 100 bacteria/neutrophil, released 97.7% of the 51Cr from labeled neutrophils. Neither a heat-killed preparation of M bovis 118F nor a live or heat-killed preparation of M bovis IBH63 (a nonhemolytic and nonpathogenic strain) induced significant (P greater than 0.05) release of 51Cr. Moraxella bovis 118F broth culture filtrates prepared for evaluation of leukocidic activity also were evaluated for hemolytic activity. These 2 toxic activities had several characteristics in common. Both were filterable, heat-labile, produced by a hemolytic strain, and were released during early logarithmic phase growth from broth cultures. Leukocidic and hemolytic activities were protected from degradation by phenylmethyl sulfonyl fluoride, a serine protease inhibitor. Leukocidic and hemolytic activities were dependent on calcium ions. Filtrate resulted in 54.1% 51Cr release from labeled neutrophils and contained 646.7 hemolytic U/ml, respectively, when saline (0.85% NaCl) + 10 mM CaCl2 solution was used as diluent. Neither saline solution nor saline + 10 mM MgCl2 solution supported leukocidic or hemolytic activity. Serum, obtained from several calves 10 to 38 days after M bovis inoculation, substantially neutralized leukocidic and hemolytic activities, compared with paired preinoculation serum samples.(ABSTRACT TRUNCATED AT 250 WORDS)

  16. The immune response of bovine mammary epithelial cells to live or heat-inactivated Mycoplasma bovis.

    Science.gov (United States)

    Zbinden, Christina; Pilo, Paola; Frey, Joachim; Bruckmaier, Rupert M; Wellnitz, Olga

    2015-09-30

    Mycoplasma bovis is an emerging bacterial agent causing bovine mastitis. Although these cell wall-free bacteria lack classical virulence factors, they are able to activate the immune system of the host. However, effects on the bovine mammary immune system are not yet well characterized and detailed knowledge would improve the prevention and therapy of mycoplasmal mastitis. The aim of this study was to investigate the immunogenic effects of M. bovis on the mammary gland in an established primary bovine mammary epithelial cell (bMEC) culture system. Primary bMEC of four different cows were challenged with live and heat-inactivated M. bovis strain JF4278 isolated from acute bovine mastitis, as well as with the type strain PG45. The immune response was evaluated 6 and 24h after mycoplasmal challenge by measuring the relative mRNA expression of selected immune factors by quantitative PCR. M. bovis triggered an immune response in bMEC, reflected by the upregulation of tumor necrosis factor-α, interleukin(IL)-1β, IL-6, IL-8, lactoferrin, Toll-like receptor-2, RANTES, and serum amyloid A mRNA. Interestingly, this cellular reaction was only observed in response to live, but not to heat-inactivated M. bovis, in contrast to other bacterial pathogens of mastitis such as Staphylococcus aureus. This study provides evidence that bMEC exhibit a strong inflammatory reaction in response to live M. bovis. The lack of a cellular response to heat-inactivated M. bovis supports the current hypothesis that mycoplasmas activate the immune system through secreted secondary metabolites.

  17. Detection of Mycobacterium tuberculosis and Mycobacterium bovis from human sputum samples through multiplex PCR.

    Science.gov (United States)

    Jabbar, Abdul; Khan, Jafar; Ullah, Aman; Rehman, Hazir; Ali, Ijaz

    2015-07-01

    Tuberculosis (TB) has a long history and being present even before the start of recording history. It has left detrimental effects on all aspect of the life and geared the developments in the science of health. TB is caused by Mycobacterium tuberculosis complex (MTBC) including five species M. tuberculosis, M. bovis, M. africanum, M. canetti, and M. microti. M. tuberculosis and M. bovis infect both animals and humans. Therefore, differentiation of these two closely related species is very important for epidemiological and management purpose. We undertook the present study to characterize mycobacteria isolated from sputum of known TB patients by conventional methods and further, by multiplex PCR (mPCR) to detect the prevalence of Zoonotic TB (TB caused by M. bovis). Sputum samples from TB patient were collected from two tertiary care hospitals in Peshawar i.e. Lady Reading Hospital and Hayatabad Medical Complex. All the samples were subjected to Ziehl Neelsen (ZN) stain, culture on Lowenstein Jensen (LJ) and Stone Brink medium, Nitrate reduction test and multiplex PCR. A total of hundred mycobacterial strains were isolated from these samples on the basis of ZN staining, cultural and biochemical methods. Later on, these isolates were subjected to multiplex PCR by using pncATB-1.2 and pncAMT-2 primers specific to M. tuberculosis and JB21, JB22 primers specific to M. bovis. By means of conventional method, these hundred cultures isolates were differentiated into M. tuberculosis (ninety six) and M. bovis (four). Furthermore, by mPCR, it was determined that out of hundred isolates, ninety-eight were identified as M. tuberculosis and two isolates as M. bovis. This molecular method enables to differentiate M. bovis from M. tuberculosis in human sputum.

  18. Toxicogenomic response of Mycobacterium bovis BCG to peracetic acid and a comparative analysis of the M. bovis BCG response to three oxidative disinfectants.

    Science.gov (United States)

    Nde, Chantal W; Toghrol, Freshteh; Jang, Hyeung-Jin; Bentley, William E

    2011-04-01

    Tuberculosis is a leading cause of death worldwide and infects thousands of Americans annually. Mycobacterium bovis causes tuberculosis in humans and several animal species. Peracetic acid is an approved tuberculocide in hospital and domestic environments. This study presents for the first time the transcriptomic changes in M. bovis BCG after treatment with 0.1 mM peracetic acid for 10 and 20 min. This study also presents for the first time a comparison among the transcriptomic responses of M. bovis BCG to three oxidative disinfectants: peracetic acid, sodium hypochlorite, and hydrogen peroxide after 10 min of treatment. Results indicate that arginine biosynthesis, virulence, and oxidative stress response genes were upregulated after both peracetic acid treatment times. Three DNA repair genes were downregulated after 10 and 20 min and cell wall component genes were upregulated after 20 min. The devR-devS signal transduction system was upregulated after 10 min, suggesting a role in the protection against peracetic acid treatment. Results also suggest that peracetic acid and sodium hypochlorite both induce the expression of the ctpF gene which is upregulated in hypoxic environments. Further, this study reveals that in M. bovis BCG, hydrogen peroxide and peracetic acid both induce the expression of katG involved in oxidative stress response and the mbtD and mbtI genes involved in iron regulation/virulence.

  19. A space-time analysis of Mycoplasma bovis: bulk tank milk antibody screening results from all Danish dairy herds in 2013-2014

    DEFF Research Database (Denmark)

    Arede, Margarida; Nielsen, Per Kantsø; Ahmed, Syed Sayeem Uddin;

    2016-01-01

    Mycoplasma bovis is an important pathogen causing severe disease outbreaks in cattle farms. Since 2011, there has been an apparent increase in M. bovis outbreaks among Danish dairy cattle herds. The dairy cattle industry performed cross-sectional antibody screening for M. bovis on four occasions,...

  20. [Mycobacterium bovis in wildlife of the dairy regions of Santa Fe (Argentina)].

    Science.gov (United States)

    Abdala, Alejandro A; Garbaccio, Sergio; Zumárraga, Martín; Tarabla, Héctor D

    2015-01-01

    Control eradication campaigns of bovine tuberculosis based on the «test and slaughter» approach were successful in many countries and regions; however, in some areas the infection persists and one of the main reasons is Mycobacterium bovis infection in wild life species. Argentina has applied the same approach since 1999, achieving progress in dairy cattle herds. Nonetheless, the wildlife role has never been investigated. The objective of this study was to determine if wildlife from the Santa Fe dairy area is infected with M. bovis. Wildlife species having a positive tuberculin skin test were captured in five dairy farms. Ninety five wildlife mammals were captured; M. bovis was recovered from 7 possums (Didelphys albiventris), from one fox (Lycolapex gimnocercus) and from one rat (Rattus norvegicus). None of the animals exhibited macroscopic lesions. The most frequently isolated M. bovis spoligotypes were types 34 (4 isolates) and 12 (3 isolates). Spoligotype 34 is the most frequently isolated type in Argentine cattle. The role of D. albiventris as spillover host of M. bovis is discussed in this study.

  1. Comparison of Sputum-Culture Conversion for Mycobacterium bovis and M. tuberculosis.

    Science.gov (United States)

    Scott, Colleen; Cavanaugh, Joseph S; Silk, Benjamin J; Ershova, Julia; Mazurek, Gerald H; LoBue, Philip A; Moonan, Patrick K

    2017-03-01

    Current US guidelines recommend longer treatment for tuberculosis (TB) caused by pyrazinamide-resistant organisms (e.g., Mycobacterium bovis) than for M. tuberculosis TB. We compared treatment response times for patients with M. bovis TB and M. tuberculosis TB reported in the United States during 2006-2013. We included culture-positive, pulmonary TB patients with genotyping results who received standard 4-drug treatment at the time of diagnosis. Time to sputum-culture conversion was defined as time between treatment start date and date of first consistently culture-negative sputum. We analyzed 297 case-patients with M. bovis TB and 30,848 case-patients with M. tuberculosis TB. After 2 months of treatment, 71% of M. bovis and 65% of M. tuberculosis TB patients showed conversion of sputum cultures to negative. Likelihood of culture conversion was higher for M. bovis than for M. tuberculosis, even after controlling for treatment administration type, sex, and a composite indicator of bacillary burden.

  2. Evaluation of Cocktails with Recombinant Proteins of Mycobacterium bovis for a Specific Diagnosis of Bovine Tuberculosis

    Directory of Open Access Journals (Sweden)

    María Laura Mon

    2014-01-01

    Full Text Available The Delayed type hypersensitivity skin test (DTH and interferon-gamma assay are used for the diagnosis of bovine tuberculosis (TBB. The specificity of these diagnoses, however, is compromised because both are based on the response against purified protein derivative of Mycobacterium bovis (PPD-B. In this study, we assessed the potential of two cocktails containing M. bovis recombinant proteins: cocktail 1 (C1: ESAT-6, CFP-10 and MPB83 and cocktail 2 (C2: ESAT-6, CFP-10, MPB83, HspX, TB10.3, and MPB70. C1, C2, and PPD-B showed similar response by DTH in M. bovis-sensitized guinea pigs. Importantly, C1 induced a lower response than PPD-B in M. avium-sensitized guinea pigs. In cattle, C1 displayed better performance than PPD-B and C2; indeed, C1 showed the least detection of animals either vaccinated or Map-infected. To optimize the composition of the cocktails, we obtained protein fractions from PPD-B and tested their immunogenicity in experimentally M. bovis-infected cattle. In one highly reactive fraction, seven proteins were identified. The inclusion of FixB in C1 enhanced the recognition of M. bovis-infected cattle without compromising specificity. Our data provide a promising basis for the future development of a cocktail for TBB detection without interference by the presence of sensitized or infected animals with other mycobacteria.

  3. Evaluation of Cocktails with Recombinant Proteins of Mycobacterium bovis for a Specific Diagnosis of Bovine Tuberculosis

    Science.gov (United States)

    Mon, María Laura; Moyano, Roberto Damián; Viale, Mariana Noelia; Colombatti Olivieri, María Alejandra; Gamietea, Ignacio José; Montenegro, Valeria Noely; Alonso, Bernardo; Santangelo, María de la Paz; Singh, Mahavir; Duran, Rosario; Romano, María Isabel

    2014-01-01

    The Delayed type hypersensitivity skin test (DTH) and interferon-gamma assay are used for the diagnosis of bovine tuberculosis (TBB). The specificity of these diagnoses, however, is compromised because both are based on the response against purified protein derivative of Mycobacterium bovis (PPD-B). In this study, we assessed the potential of two cocktails containing M. bovis recombinant proteins: cocktail 1 (C1): ESAT-6, CFP-10 and MPB83 and cocktail 2 (C2): ESAT-6, CFP-10, MPB83, HspX, TB10.3, and MPB70. C1, C2, and PPD-B showed similar response by DTH in M. bovis-sensitized guinea pigs. Importantly, C1 induced a lower response than PPD-B in M. avium-sensitized guinea pigs. In cattle, C1 displayed better performance than PPD-B and C2; indeed, C1 showed the least detection of animals either vaccinated or Map-infected. To optimize the composition of the cocktails, we obtained protein fractions from PPD-B and tested their immunogenicity in experimentally M. bovis-infected cattle. In one highly reactive fraction, seven proteins were identified. The inclusion of FixB in C1 enhanced the recognition of M. bovis-infected cattle without compromising specificity. Our data provide a promising basis for the future development of a cocktail for TBB detection without interference by the presence of sensitized or infected animals with other mycobacteria. PMID:25110654

  4. Isolation of Mycobacterium bovis from Free-Ranging Wildlife in South Korea.

    Science.gov (United States)

    Jang, Yunho; Ryoo, Soyoon; Lee, Hyunkyoung; Kim, Narae; Lee, Hang; Park, So-Young; Song, Woong-Seog; Kim, Jong-Taek; Lee, Hee Soo; Myung Kim, Jae

    2017-01-01

    We demonstrate Mycobacterium bovis infection in wild boar ( Sus scrofa ) in South Korea. During 2012-15, we attempted to isolate M. bovis from 847 wild animals, mainly Korean water deer ( Hydropotes inermis argyropus), raccoon dogs ( Nyctereutes procyonoides ), and wild boar, from 11 regions in South Korea. We isolated M. bovis from three of 118 wild boar (2.5%) captured in Gyeonggi Province, where bovine tuberculosis (bTB) outbreaks have also occurred in livestock. Spoligotypes and mycobacterial interspersed repetitive units-variable number tandem repeats types of these M. bovis isolates (SB0140 and SB1040, 4-2-3-3-7-5-5-4-4-3-4-3 and 5-2-3-3-7-5-5-4-3-10-5-2; MIRU4, MIRU16, MIRU27, MIRU31, ETR-A, ETR-B, ETR-C, QUB11b, QUB26, QUB3336, VNTR2401, and VNTR3171) have also been identified from farmed livestock such as cattle ( Bos taurus coreanae), Formosan sika deer ( Cervus nippon taiouanus), and American elk ( Cervus canadensis ) in the country. In South Korea, bTB appears to be endemic in livestock, and there are numerous opportunities for contact between wild boar and livestock due to high population densities and broad activity ranges. Our results support the hypothesis that M. bovis is transmitted between domestic and wild animals.

  5. Phylogenetic analysis of Mexican Babesia bovis isolates using msa and ssrRNA gene sequences.

    Science.gov (United States)

    Genis, Alma D; Mosqueda, Juan J; Borgonio, Verónica M; Falcón, Alfonso; Alvarez, Antonio; Camacho, Minerva; de Lourdes Muñoz, Maria; Figueroa, Julio V

    2008-12-01

    Variable merozoite surface antigens of Babesia bovis are exposed glycoproteins having a role in erythrocyte invasion. Members of this gene family include msa-1 and msa-2 (msa-2c, msa-2a(1), msa-2a(2), and msa-2b). Small subunit ribosomal (ssr)RNA gene is subject to evolutive pressure and has been used in phylogenetic studies. To determine the phylogenetic relationship among B. bovis Mexican isolates using different genetic markers, PCR amplicons, corresponding to msa-1, msa-2c, msa-2b, and ssrRNA genes, were cloned and plasmids carrying the corresponding inserts were sequenced. Comparative analysis of nucleotide and deduced amino acid sequences revealed distinct degrees of variability and identity among the coding gene sequences obtained from 12 geographically different B. bovis isolates and a reference strain. Overall sequence identities of 47.7%, 72.3%, 87.7%, and 94% were determined for msa-1, msa-2b, msa-2c, and ssrRNA, respectively. A robust phylogenetic tree was obtained with msa-2b sequences. The phylogenetic analysis suggests that Mexican B. bovis isolates group in clades not concordant with the Mexican geography. However, the Mexican isolates group together in an American clade separated from the Australian clade. Sequence heterogeneity in msa-1, msa-2b, and msa-2c coding regions of Mexican B. bovis isolates present in different geographical regions can be a result of either differential evolutive pressure or cattle movement from commercial trade.

  6. ELISA-based serological survey of Mycoplasma bovis in cattle in three local government areas in Adamawa State, Nigeria

    Directory of Open Access Journals (Sweden)

    Markus Isa Francis

    2015-06-01

    Full Text Available A serological survey for the detection of antibodies to Mycoplasma bovis was conducted in Adamawa State, Nigeria during the year 2012. A total of 400 serum samples were collected from three local government areas (LGA namely Yola (n=140, Mubi (n=130 and Ganye (n=130, and the samples were examined for the presence of M. bovis antibodies using BIO-X M. bovis antibody ELISA Kit. The overall seroprevalence of M. bovis was recorded as 19.5% (n=78/400. The highest seroprevalence was recorded in Ganye LGA (27.7%; n=36/130, followed by Yola LGA (20.0%; n=28/140 and Mubi LGA (10.8%; n=14/130. The cattle aging <1-year had the highest prevalence (25.8%, followed by 4-year-old cattle (22.4%. Based on sex distribution, 20.4% of the cows and 17.4% of the bulls were seropositive to M. bovis. Breed susceptibility to M. bovis infection showed that White Fulani had the highest prevalence (21.8%, and Sokoto Gudali had the lowest (11.9%. No statistical significant association was found between M. bovis infection and age, sex and breed of the cattle. In conclusion, the above findings are indicative for the presence of M. bovis in the study area.

  7. 76 FR 14289 - Bacillus thuringiensis

    Science.gov (United States)

    2011-03-16

    ... AGENCY 40 CFR Part 174 Bacillus thuringiensis eCry3.1Ab Protein in Corn; Temporary Exemption From the... regulation extends a temporary exemption from the requirement of a tolerance for residues of Bacillus... permissible level for residues of Bacillus thuringiensis eCry3.1Ab protein in corn. The temporary...

  8. 75 FR 34040 - Bacillus thuringiensis

    Science.gov (United States)

    2010-06-16

    ... AGENCY 40 CFR Part 174 Bacillus thuringiensis eCry3.1Ab Protein in Corn; Temporary Exemption from the... regulation establishes a temporary exemption from the requirement of a tolerance for residues of Bacillus... Bacillus thuringiensis eCry3.1Ab protein in corn under the FFDCA. The temporary tolerance exemption...

  9. Natural Babesia bovis infection in water buffaloes (Bubalus bubalis) and crossbred cattle under field conditions in Egypt

    DEFF Research Database (Denmark)

    Mahmmod, Yasser

    2014-01-01

    Background: There is a little or no data available on the natural Babesia bovis (B. bovis) infection in water buffaloes (Bubalus bubalis) comparing to the available one for cattle. This study was conducted to investigate the natural B. bovis infection in water buffaloes in comparison to crossbred...... of cattle showed dark brown to dark red (coffee-color) urine, hemoglobinuria and nervous manifestations while these manifestations were not detected in the infected buffaloes. Hematological changes in both species however, these changes were less significant in buffaloes than those reported in cattle....... Conclusion: This paper documents the first description of natural B. bovis infection in water buffaloes which were found to be more likely to be tolerant than cattle to the natural clinical infection with B. bovis and its subsequent haematological changes. Our finding may lead to a better understanding...

  10. [Evaluation of variable number of tandem repeats (VNTR) isolates of Mycobacterium bovis in Algeria].

    Science.gov (United States)

    Sahraoui, Naima; Muller, Borna; Djamel, Yala; Fadéla, Boulahbal; Rachid, Ouzrout; Jakob, Zinsstag; Djamel, Guetarni

    2010-01-01

    The discriminatory potency of variable number of tandem repeats (VNTR), based on 7 loci (MIRU 26, 27 and 5 ETRs A, B, C, D, E) was assayed on Mycobacterium bovis strains obtained from samples due to tuberculosis in two slaughterhouses in Algeria. The technique of MIRU-VNTR has been evaluated on 88 strains of M. bovis and one strain of M. caprea and shows 41 different profiles. Results showed that the VNTR were highly discriminatory with an allelic diversity of 0.930 when four loci (ETR A, B, C and MIRU 27) were highly discriminatory (h>0.25) and three loci (ETR D and E MIRU 26) moderately discriminatory (0.11bovis in Algeria. The VNTR technique has proved a valuable tool for further development and application of epidemiological research for the of tuberculosis transmission in Algeria.

  11. Detection of Mycobacterium bovis and Mycobacterium tuberculosis from Cattle: Possible Public Health Relevance

    DEFF Research Database (Denmark)

    Thakur, Aneesh; Sharma, Mandeep; Katoch, Vipin C.;

    2012-01-01

    Mycobacterium bovis and Mycobacterium tuberculosis infect both animals and humans. The disease epidemiology by these agents differs in developed and developing countries due to the differences in the implementation of the prevention and control strategies. The present study describes the detection...... of M. bovis and M. tuberculosis from specimens of lungs and pulmonary lymph nodes of four cattle died in an organized herd of 183 cattle in the state of Himachal Pradesh, India, with inconclusive skin test results. Identification and distinction of these closely related mycobacterial species was done...... by PCR-RFLP targeting hsp65 gene followed by spacer oligonucleotide typing. Mixed infection of M. bovis and M. tuberculosis was detected in one cattle....

  12. Bovine infectious keratoconjunctivitis: carrier state of Moraxella bovis and the development of preventive measures against disease.

    Science.gov (United States)

    Pugh, G W; Hughes, D E

    1975-08-15

    Bovine infectious keratoconjunctivitis, which is caused by Moraxella bovis, occurs perennially in all cattle-raising areas. Disease may occur any time during the year but manifests itself often during summer when disease-enhancing factors such as ultraviolet radiation and fly population are increased. Although some recent research findings indicate that cattle can be immunized against the disease, many problems have to be overcome before the cattle industry can benefit from such immunization. Some pressing problems relate to lack of cross-protection between vaccines of different strains of M bovis, resistant or resilient carrier states, and how these factors affect vaccination regimens undertaken. Though cattle vaccinated with 1 strain are somewhat protected against subsequent challange by the same strain, protection against heterologous strains has not been demonstrated. In most herds, cattle harbor M bovis, and these carriers may serve as a source of infection.

  13. Filamentous-haemagglutinin-like protein genes encoded on a plasmid of Moraxella bovis.

    Science.gov (United States)

    Kakuda, Tsutomu; Sarataphan, Nopporn; Tanaka, Tetsuya; Takai, Shinji

    2006-11-26

    The complete nucleotide sequence of a plasmid, pMBO-1, from Moraxella bovis strain Epp63 was determined. We identified 30 open reading frames (ORFs) encoded by the 44,215bp molecule. Two large ORFs, flpA and flpB, encoding proteins with similarity to Bordetella pertussis filamentous haemagglutinin (FHA), were identified on the same plasmid. The gene for a specific accessory protein (Fap), which may play a role in the secretion of Flp protein, was also identified. Reverse transcriptase PCR analysis of total RNA isolated from M. bovis Epp63 indicated that the flpA, flpB, and fap genes are all transcribed. Southern blot analysis indicated that the flp and fap genes are present in other clinical isolates of geographically diverse M. bovis.

  14. Identification and characterisation of a biosynthetic locus for Moraxella bovis lipo-oligosaccharide.

    Science.gov (United States)

    Faglin, Isabelle; Grice, I Darren; Ratnayake, S R A M Eranda; Daal, Terese-Marie; Singh, Sanjesh; Wilson, Jennifer C; Peak, Ian R

    2016-02-08

    Moraxella bovis is a Gram-negative gammaproteobacterium and is one of the causative agents of infectious bovine keratoconjunctivitis. The structure of lipooligosaccharide (LOS) from strain Epp63 was recently elucidated. In the present study a genetic locus of seven encoding genes with high similarity to glycosyltransferases has been identified. Mutation of these putative glycosyltransferase genes resulted in M. bovis mutant bacteria that expressed truncated LOS structures. The structures of the oligosaccharide (OS) expressed by the mutant strains were elucidated and demonstrated the role of the glycosyltransferase enzymes in the LOS biosynthesis of M. bovis. The glycosyltransferase genes designated lgt1, lgt3, and lgt6 are highly similar to the genes in the related bacterium M. catarrhalis. In addition, there are syntenic similarities with the corresponding LOS biosynthesis locus in M. catarrhalis and other members of Moraxellaceae.

  15. Molecular detection and identification of Babesia bovis and Babesia bigemina in cattle in northern Thailand.

    Science.gov (United States)

    Cao, Shinuo; Aboge, Gabriel Oluga; Terkawi, Mohamad Alaa; Yu, Longzheng; Kamyingkird, Ketsarin; Luo, Yuzi; Li, Yan; Goo, Youn-Kyoung; Yamagishi, Junya; Nishikawa, Yoshifumi; Yokoyama, Naoaki; Suzuki, Hiroshi; Igarashi, Ikuo; Maeda, Ryuichiro; Inpankaew, Tawin; Jittapalapong, Sathaporn; Xuan, Xuenan

    2012-09-01

    Although Babesia bovis and Babesia bigemina infections cause economic losses in the cattle industry in northern Thailand, there is inadequate information on Babesia isolates present in the area. Therefore, to determine the prevalence and genetic relationship between Babesia isolates, we screened 200 blood samples of cattle from Chiang Rai, Chiang Mai, and Lumpang provinces of northern Thailand. A nested polymerase chain reaction using primers targeting B. bovis spherical body protein 2 (BboSBP2) and B. bigemina rhoptry-associated protein 1a (BbiRAP-1a) genes revealed a prevalence of 12 and 21 % for B. bovis and B. bigemina, respectively, while that of mixed infections was 6.5 % samples. The prevalences of B. bovis in Chiang Rai, Chiang Mai, and Lumpang were 9.5, 3.7, and 25.5 %, respectively. For B. bigemina, the prevalences were 15.8, 12.9, and 39.2 % in Chiang Rai, Chiang Mai, and Lumpang, respectively. Mixed infections with B. bovis and B. bigemina were 6.3 % in Chiang Rai, 1.9 % in Chiang Mai, and 13.7 % in Lumpang. The identical sequences of either BboSBP2 gene or BbiRAP-1a gene were shared among the Babesia isolates in the three provinces of northern Thailand. Further analysis using the internal transcribed spacer gene revealed at least four genotypes for B. bovis and five genotypes for B. bigemina in northern Thailand, while the sequences present great genetic diversities in the different isolates. Overall, we have demonstrated a high prevalence and polymorphism of Babesia parasites in northern Thailand calling for the need to design effective control programs for bovine babesiosis.

  16. Asymptomatic cattle naturally infected with Mycobacterium bovis present exacerbated tissue pathology and bacterial dissemination.

    Directory of Open Access Journals (Sweden)

    Álvaro Menin

    Full Text Available Rational discovery of novel immunodiagnostic and vaccine candidate antigens to control bovine tuberculosis (bTB requires knowledge of disease immunopathogenesis. However, there remains a paucity of information on the Mycobacterium bovis-host immune interactions during the natural infection. Analysis of 247 naturally PPD+ M. bovis-infected cattle revealed that 92% (n = 228 of these animals were found to display no clinical signs, but presented severe as well as disseminated bTB-lesions at post-mortem examination. Moreover, dissemination of bTB-lesions positively correlated with both pathology severity score (Spearman r = 0.48; p<0.0001 and viable tissue bacterial loads (Spearman r = 0.58; p = 0.0001. Additionally, granuloma encapsulation negatively correlated with M. bovis growth as well as pathology severity, suggesting that encapsulation is an effective mechanism to control bacterial proliferation during natural infection. Moreover, multinucleated giant cell numbers were found to negatively correlate with bacterial counts (Spearman r = 0.25; p = 0.03 in lung granulomas. In contrast, neutrophil numbers in the granuloma were associated with increased M. bovis proliferation (Spearman r = 0.27; p = 0.021. Together, our findings suggest that encapsulation and multinucleated giant cells control M. bovis viability, whereas neutrophils may serve as a cellular biomarker of bacterial proliferation during natural infection. These data integrate host granuloma responses with mycobacterial dissemination and could provide useful immunopathological-based biomarkers of disease severity in natural infection with M. bovis, an important cattle pathogen.

  17. Genetic diversity and antigenicity variation of Babesia bovis merozoite surface antigen-1 (MSA-1) in Thailand.

    Science.gov (United States)

    Tattiyapong, Muncharee; Sivakumar, Thillaiampalam; Takemae, Hitoshi; Simking, Pacharathon; Jittapalapong, Sathaporn; Igarashi, Ikuo; Yokoyama, Naoaki

    2016-07-01

    Babesia bovis, an intraerythrocytic protozoan parasite, causes severe clinical disease in cattle worldwide. The genetic diversity of parasite antigens often results in different immune profiles in infected animals, hindering efforts to develop immune control methodologies against the B. bovis infection. In this study, we analyzed the genetic diversity of the merozoite surface antigen-1 (msa-1) gene using 162 B. bovis-positive blood DNA samples sourced from cattle populations reared in different geographical regions of Thailand. The identity scores shared among 93 msa-1 gene sequences isolated by PCR amplification were 43.5-100%, and the similarity values among the translated amino acid sequences were 42.8-100%. Of 23 total clades detected in our phylogenetic analysis, Thai msa-1 gene sequences occurred in 18 clades; seven among them were composed of sequences exclusively from Thailand. To investigate differential antigenicity of isolated MSA-1 proteins, we expressed and purified eight recombinant MSA-1 (rMSA-1) proteins, including an rMSA-1 from B. bovis Texas (T2Bo) strain and seven rMSA-1 proteins based on the Thai msa-1 sequences. When these antigens were analyzed in a western blot assay, anti-T2Bo cattle serum strongly reacted with the rMSA-1 from T2Bo, as well as with three other rMSA-1 proteins that shared 54.9-68.4% sequence similarity with T2Bo MSA-1. In contrast, no or weak reactivity was observed for the remaining rMSA-1 proteins, which shared low sequence similarity (35.0-39.7%) with T2Bo MSA-1. While demonstrating the high genetic diversity of the B. bovis msa-1 gene in Thailand, the present findings suggest that the genetic diversity results in antigenicity variations among the MSA-1 antigens of B. bovis in Thailand.

  18. Genome sequence of Babesia bovis and comparative analysis of apicomplexan hemoprotozoa.

    Directory of Open Access Journals (Sweden)

    Kelly A Brayton

    2007-10-01

    Full Text Available Babesia bovis is an apicomplexan tick-transmitted pathogen of cattle imposing a global risk and severe constraints to livestock health and economic development. The complete genome sequence was undertaken to facilitate vaccine antigen discovery, and to allow for comparative analysis with the related apicomplexan hemoprotozoa Theileria parva and Plasmodium falciparum. At 8.2 Mbp, the B. bovis genome is similar in size to that of Theileria spp. Structural features of the B. bovis and T. parva genomes are remarkably similar, and extensive synteny is present despite several chromosomal rearrangements. In contrast, B. bovis and P. falciparum, which have similar clinical and pathological features, have major differences in genome size, chromosome number, and gene complement. Chromosomal synteny with P. falciparum is limited to microregions. The B. bovis genome sequence has allowed wide scale analyses of the polymorphic variant erythrocyte surface antigen protein (ves1 gene family that, similar to the P. falciparum var genes, is postulated to play a role in cytoadhesion, sequestration, and immune evasion. The approximately 150 ves1 genes are found in clusters that are distributed throughout each chromosome, with an increased concentration adjacent to a physical gap on chromosome 1 that contains multiple ves1-like sequences. ves1 clusters are frequently linked to a novel family of variant genes termed smorfs that may themselves contribute to immune evasion, may play a role in variant erythrocyte surface antigen protein biology, or both. Initial expression analysis of ves1 and smorf genes indicates coincident transcription of multiple variants. B. bovis displays a limited metabolic potential, with numerous missing pathways, including two pathways previously described for the P. falciparum apicoplast. This reduced metabolic potential is reflected in the B. bovis apicoplast, which appears to have fewer nuclear genes targeted to it than other apicoplast

  19. Differences in the antimicrobial susceptibility profiles of Moraxella bovis, M. bovoculi and M. ovis

    Science.gov (United States)

    Maboni, Grazieli; Gressler, Leticia T.; Espindola, Julia P.; Schwab, Marcelo; Tasca, Caiane; Potter, Luciana; de Vargas, Agueda Castagna

    2015-01-01

    The aim of this study was to determine the differences in the antimicrobial susceptibility profiles of Moraxella bovis, M. bovoculi and M. ovis. Thirty-two strains of Moraxella spp. isolated from cattle and sheep with infectious keratoconjunctivitis were tested via broth microdilution method to determine their susceptibility to ampicillin, cefoperazone, ceftiofur, cloxacillin, enrofloxacin, florfenicol, gentamicin, neomycin, oxytetracycline and penicillin. The results demonstrated that Moraxella spp. strains could be considered sensitive for most of the antimicrobials tested in this study, but differences between the antimicrobial susceptibility profiles of these three Moraxella species were found. M. bovis might differ from other species due to the higher MIC and MBC values it presented. PMID:26273272

  20. [Isolation of Chlamydia psittaci and Moraxella bovis from infectious keratoconjunctivitis in lambs].

    Science.gov (United States)

    Trávnícek, M; Dravecký, T; Balascák, J

    1982-08-01

    Out of 189 lambs in the flock, 25 animals suffered from bilateral or unilateral conjunctivitis, or keratoconjunctivitis. By serological examination (RVK), positive levels of antibodies to the group-specific antigen of Chl. psittaci were found in three out of six lambs examined by laboratory methods. Bacteriological examination of eye smears of six lambs showed in four cases the infection by microorganisms of Moraxella bovis. Smears from the conjunctivas of these lambs were after preparation instilled in the yolk sacs of six to seven days old chicken embryos. One strain of Chlamydia psittaci was isolated from the same material as Moraxella bovis.

  1. Bacillus velezensis is not a later heterotypic synonym of Bacillus amyloliquefaciens; Bacillus methylotrophicus, Bacillus amyloliquefaciens subsp plantarum and ‘Bacillus oryzicola’ are later heterotypic synonyms of Bacillus

    Science.gov (United States)

    The rhizosphere isolated bacteria belonging to the Bacillus amyloliquefaciens subsp. plantarum and Bacillus methylotrophicus clades are an important group of strains that are used as plant growth promoters and antagonists of plant pathogens. These properties have made these strains the focus of comm...

  2. Essential Bacillus subtilis genes

    DEFF Research Database (Denmark)

    Kobayashi, K.; Ehrlich, S.D.; Albertini, A.

    2003-01-01

    To estimate the minimal gene set required to sustain bacterial life in nutritious conditions, we carried out a systematic inactivation of Bacillus subtilis genes. Among approximate to4,100 genes of the organism, only 192 were shown to be indispensable by this or previous work. Another 79 genes were...

  3. Characterization of Bacillus cereus

    NARCIS (Netherlands)

    Wijnands LM; Dufrenne JB; Leusden FM; MGB

    2002-01-01

    Bacillus cereus is a ubiquitary microorganism that may cause food borne disease. Pathogenicity, however, depends on various characteristics such as the ability to form (entero)-toxin(s) that can not be detected by microbiological methods. Further characterization of pathogenic properties is not only

  4. Biodiversity in Bacillus cereus

    NARCIS (Netherlands)

    Pielaat A; Fricker M; Nauta MJ; Leusden FM van; MGB

    2006-01-01

    Experiments have been performed by different partners to identify variability in properties of Bacillus cereus strains that contribute to the extent of their virulence as part of an EU project. To this end, 100 B. cereus strains were selected and screened for biological properties, such as toxin pro

  5. Draft Genome Sequences of Three Alkaliphilic Bacillus Strains, Bacillus wakoensis JCM 9140T, Bacillus akibai JCM 9157T, and Bacillus hemicellulosilyticus JCM 9152T

    OpenAIRE

    Yuki, Masahiro; Oshima, Kenshiro; Suda, Wataru; OSHIDA, Yumi; Kitamura, Keiko; Iida, Toshiya; Hattori, Masahira; Ohkuma, Moriya

    2014-01-01

    Here, we report the draft genome sequences of the type strains of three cellulolytic or hemicellulolytic alkaliphilic Bacillus species: Bacillus wakoensis, Bacillus akibai, and Bacillus hemicellulosilyticus. The genome information for these three strains will be useful for studies of alkaliphilic Bacillus species, their evolution, and biotechnological applications for their enzymes.

  6. Genome analysis shows Bacillus axarquiensis is not a later heterotypic synonym of Bacillus mojavensis; Reclassification of Bacillus malacitensis and Brevibacterium halotolerans as heterotypic synonyms of Bacillus axarquiensis

    Science.gov (United States)

    Bacillus axarquiensis and Bacillus malacitensis were previously reported to be later heterotypic synonyms of Bacillus mojavensis, based primarily on DNA-DNA relatedness values. We have sequenced draft genomes of Bacillus axarquiensis NRRL B-41617**T and Bacillus malacitensis NRRL B-41618**T. Compara...

  7. Nano-Mechanical Properties of Heat Inactivated Bacillus anthracis and Bacillus thuringiensis Spores

    Science.gov (United States)

    2008-03-01

    NANO-MECHANICAL PROPERTIES OF HEAT INACTIVATED BACILLUS ANTHRACIS AND BACILLUS THURINGIENSIS ...GAP/ENP/08-M07 NANO-MECHANICAL PROPERTIES OF HEAT INACTIVATED BACILLUS ANTHRACIS AND BACILLUS THURINGIENSIS SPORES THESIS...AFIT/GAP/ENP/08-M07 NANO-MECHANICAL PROPERTIES OF HEAT INACTIVATED BACILLUS ANTHRACIS AND BACILLUS THURINGIENSIS SPORES Jessica

  8. Chemical composition and nutrient degradability in elephant grass silage inoculated with Streptococcus bovis isolated from the rumen.

    Science.gov (United States)

    Ferreira, Daniele J; Zanine, Anderson M; Lana, Rogério P; Ribeiro, Marinaldo D; Alves, Guilherme R; Mantovani, Hilário C

    2014-03-01

    The objective of the present study was to assess the chemical and bromatological composition and in situ degradability of elephant grass silages inoculated with Streptococcus bovis isolated from cattle rumen. A complete randomized design was used with four treatments and six replications: elephant grass silage, elephant grass silage inoculated with 10(6) CFU/g Streptococcus bovis JB1 strains; elephant grass silage inoculated with 106 CFU/g Streptococcus bovis HC5 strains; elephant grass silage inoculated with 106 CFU/g Enterococcus faecium with six replications each. The pH and ammoniacal nitrogen values were lower (P<0.05) for the silages inoculated with Streptococcus bovis JB1 and HC5, respectively. The silage inoculated with Streptococcus bovis had a higher crude protein content (P<0.05) and there were no differences for the fiber contents in the silage. The (a)soluble fraction degradability, especially in the silages inoculated with Streptococcus bovis JB1 and HC5, had higher values, 30.77 and 29.97%, for dry matter and 31.01 and 36.66% for crude protein, respectively. Inoculation with Streptococcus bovis improved the fermentation profile, protein value and rumen degradability of the nutrients.

  9. Evaluation of pathogenesis caused in cattle and guinea pig by a Mycobacterium bovis strain isolated from wild boar

    Directory of Open Access Journals (Sweden)

    Di Rienzo Julio

    2011-07-01

    Full Text Available Abstract Background In many regions of the world, wild mammals act as reservoir of Mycobacterium bovis, a situation that prevents the eradication of bovine tuberculosis. In order to observe whether a strain isolated from a wild boar, previously tested as highly virulent in a mice model, is also virulent in cattle, we performed cattle experimental inoculation with this strain Results Groups of Friesian calves were either infected with the wild boar strain M. bovis 04-303 or with the bovine strain NCTC10772 as a control. We found that antigen-specific IFN-γ release in whole blood samples occurred earlier in animals infected with M. bovis 04-303. Both M. bovis strains resulted in a positive skin test, with animals infected with the wild boar isolate showing a stronger response. These results and the presence of more severe organ lesions, with granuloma and pneumonic areas in cattle demonstrate that the wild boar isolate is more virulent than the NCTC10772 strain. Additionally, we tested the infectivity of the M. bovis strains in guinea pigs and found that M. bovis 04-303 had the highest pathogenicity. Conclusions M. bovis strains isolated from wild boars may be pathogenic for cattle, producing TB lesions.

  10. Identification of Moraxella bovis and related species from calves with IBK and goats by qualitative genetic transformation assay.

    Science.gov (United States)

    Kodjo, A; Exbrayat, P; Richard, Y

    1994-07-01

    Eight Moraxella bovis strains isolated from bovine pink eye, 16 Moraxella bovis related strains isolated from healthy goats nasal flora, one nonhemolytic Moraxella sp. isolated from bovine conjunctivitis and different collection strains of Moraxella and Branhamella genus were studied through the combined use of qualitative genetic transformation assay and the ability to grow on a moraxella bovis defined medium (medium MB). Crude DNA extracted from the strains studied were used to transform two mutant auxotroph competent strains of Moraxella bovis CIP 103741 and CIP 103743. Non-specific positive transformation was obtained with mutant assay strain CIP 103743 when treated with DNA from caprine or bovine Moraxella strains, collection strains of Moraxella bovis, Moraxella lacunata, Moraxella nonliquefaciens and strains of Branhamella genus, whereas specific transformation was observed with mutant assay CIP 103741 when treated only with DNA from all the Moraxella bovis (collection and clinical isolates) and 14 of the 16 caprine Moraxella bovis related strains. The specificity and the simplicity of the test make it suitable for use in clinical laboratories.

  11. Gut transcriptome of replete adult female cattle ticks, Rhipicephalus (Boophilus) microplus, feeding upon a Babesia bovis-infected bovine host.

    Science.gov (United States)

    Heekin, Andrew M; Guerrero, Felix D; Bendele, Kylie G; Saldivar, Leo; Scoles, Glen A; Dowd, Scot E; Gondro, Cedric; Nene, Vishvanath; Djikeng, Appolinaire; Brayton, Kelly A

    2013-09-01

    As it feeds upon cattle, Rhipicephalus (Boophilus) microplus is capable of transmitting a number of pathogenic organisms, including the apicomplexan hemoparasite Babesia bovis, a causative agent of bovine babesiosis. The R. microplus female gut transcriptome was studied for two cohorts: adult females feeding on a bovine host infected with B. bovis and adult females feeding on an uninfected bovine. RNA was purified and used to generate a subtracted cDNA library from B. bovis-infected female gut, and 4,077 expressed sequence tags (ESTs) were sequenced. Gene expression was also measured by a microarray designed from the publicly available R. microplus gene index: BmiGI Version 2. We compared gene expression in the tick gut from females feeding upon an uninfected bovine to gene expression in tick gut from females feeding upon a splenectomized bovine infected with B. bovis. Thirty-three ESTs represented on the microarray were expressed at a higher level in female gut samples from the ticks feeding upon a B. bovis-infected calf compared to expression levels in female gut samples from ticks feeding on an uninfected calf. Forty-three transcripts were expressed at a lower level in the ticks feeding upon B. bovis-infected female guts compared with expression in female gut samples from ticks feeding on the uninfected calf. These array data were used as initial characterization of gene expression associated with the infection of R. microplus by B. bovis.

  12. Effects of inulin chain length on fermentation by equine fecal bacteria and Streptococcus bovis

    Science.gov (United States)

    Fructans from pasture can be fermented by Gram-positive bacteria (e.g., Streptococcus bovis) in the equine hindgut, increasing production of lactic acid and decreasing pH. The degree of polymerization (DP) of fructans has been suggested to influence fermentation rates. The objective of the current ...

  13. Mycobacterium bovis in a Free-Ranging Black Rhinoceros, Kruger National Park, South Africa, 2016

    Science.gov (United States)

    Buss, Peter E.; van Helden, Paul D.; Parsons, Sven D.C.

    2017-01-01

    In 2016, an emaciated black rhinoceros (Diceros bicornis) was found in Kruger National Park, South Africa. An interferon-γ response was detected against mycobacterial antigens, and lung tissue was positive for Mycobacterium bovis. This case highlights the risk that tuberculosis presents to rhinoceros in M. bovis–endemic areas. PMID:28221132

  14. Transfected babesia bovis expressing a tick GST as a live vector vaccine

    Science.gov (United States)

    The Rhipicephalus microplus tick is a notorious blood-feeding ectoparasite of livestock, especially cattle, responsible for massive losses in animal production. It is the main vector for transmission of pathogenic bacteria and parasites, including Babesia bovis, an intraerythrocytic apicomplexan pro...

  15. Mycobacterium bovis infection in the lion (Panthera leo): Current knowledge, conundrums and research challenges.

    Science.gov (United States)

    Viljoen, Ignatius M; van Helden, Paul D; Millar, Robert P

    2015-06-12

    Mycobacterium bovis has global public-health and socio-economic significance and can infect a wide range of species including the lion (Panthera leo) resulting in tuberculosis. Lions are classified as vulnerable under the IUCN Red List of Threatened Species and have experienced a 30% population decline in the past two decades. However, no attempt has been made to collate and critically evaluate the available knowledge of M. bovis infections in lions and potential effects on population. In this review we set out to redress this. Arguments suggesting that ingestion of infected prey animals are the main route of infection for lions have not been scientifically proven and research is needed into other possible sources and routes of infection. The paucity of knowledge on host susceptibility, transmission directions and therefore host status, manifestation of pathology, and epidemiology of the disease in lions also needs to be addressed. Advances have been made in diagnosing the presence of M. bovis in lions. However, these diagnostic tests are unable to differentiate between exposure, presence of infection, or stage of disease. Furthermore, there are contradictory reports on the effects of M. bovis on lion populations with more data needed on disease dynamics versus the lion population's reproductive dynamics. Knowledge on disease effects on the lion reproduction and how additional stressors such as drought or co-morbidities may interact with tuberculosis is also lacking. Filling these knowledge gaps will contribute to the understanding of mycobacterial infections and disease in captive and wild lions and assist in lion conservation endeavours.

  16. Virulence of two strains of Mycobacterium bovis in cattle following aerosol infection

    Science.gov (United States)

    Background Over the past two decades, highly virulent strains of Mycobacterium tuberculosis have emerged and spread rapidly in humans, suggesting a selective advantage based upon virulence. A similar scenario has not been described for Mycobacterium bovis infection in cattle (i.e., Bovine Tuberculos...

  17. Relative virulence in bison and cattle of bison-associated genotypes of Mycoplasma bovis

    Science.gov (United States)

    Background. Mycoplasma bovis is a cause of respiratory disease in cattle and the bacterium most frequently isolated from bovine respiratory disease complex. It has recently emerged as a major health problem in bison, causing pharyngitis, pneumonia, arthritis, dystocia and abortion. In cattle, M. b...

  18. Molecular epidemiological analysis of Mycoplasma bovis isolates from the United Kingdom shows two genetically distinct clusters

    DEFF Research Database (Denmark)

    McAuliffe, Laura; Kokotovic, Branko; Ayling, Roger D.;

    2004-01-01

    Mycoplasma bovis is an important veterinary pathogen causing pneumonia, arthritis, and mastitis in infected cattle. We investigated the genetic diversity of 53 isolates collected in the United Kingdom between 1996 and 2002 with pulsed-field gel electrophoresis (PFGE), amplified fragment length...

  19. Antigen-Specific IP-10 Release Is a Sensitive Biomarker of Mycobacterium bovis Infection in Cattle.

    Directory of Open Access Journals (Sweden)

    Sven D C Parsons

    Full Text Available The most widely used ante-mortem diagnostic tests for tuberculosis in cattle are the tuberculin skin test and the interferon-gamma (IFN-γ release assay, both of which measure cell-mediated immune responses to Mycobacterium bovis infection. However, limitations in the performance of these tests results in a failure to identify all infected animals. In attempting to increase the range of diagnostic tests for tuberculosis, measurement of the cytokine IP-10 in antigen-stimulated blood has previously been shown to improve the detection of M. tuberculosis and M. bovis infection, in humans and African buffaloes (Syncerus caffer, respectively. In the present study, 60 cattle were identified by the single intradermal comparative tuberculin test as tuberculosis reactors (n = 24 or non-reactors (n = 36 and the release of IFN-γ and IP-10 in antigen-stimulated whole blood from these animals was measured using bovine specific ELISAs. There was a strong correlation between IP-10 and IFN-γ production in these samples. Moreover, measurement of the differential release of IP-10 in response to stimulation with M. bovis purified protein derivative (PPD and M. avium PPD distinguished between reactor and non-reactor cattle with a sensitivity of 100% (95% CI, 86%-100% and a specificity of 97% (95% CI, 85%-100%. These results suggest that IP-10 might prove valuable as a diagnostic biomarker of M. bovis infection in cattle.

  20. Development of a tandem repeat-based multilocus typing system distinguishing Babesia bovis geographic isolates

    Science.gov (United States)

    Mini and microsatellite sequences have proven to be excellent tools for the differentiation of strains and populations in several protozoan parasites due to their high variability. In the present work we have searched the genome of the tick-transmitted bovine hemoprotozoon Babesia bovis for tandem r...

  1. Association of microRNAs with antibody response to mycoplasma bovis in beef cattle

    Science.gov (United States)

    The objective of this study was to identify microRNAs associated with a serum antibody response to Mycoplasma bovis in beef cattle. Serum from sixteen beef calves was collected at three points: in summer after calves were born, in fall at weaning, and in the following spring. All sera collected in t...

  2. Serologic response of vaccinated cattle to strains of Moraxella bovis isolated during epizootics of keratoconjunctivitis.

    Science.gov (United States)

    Pugh, G W; Hughes, D E; Booth, G D

    1978-01-01

    In studies to determine whether there were antigenic differences between strains (isolates) of Moraxella bovis, the sera from vaccinated calves were tested with isolates of M bovis while the calves were experiencing epizootics of infectious bovine keratoconjunctivitis (IBK). Before the epizootics of IBK, the calves were intramuscularly vaccinated with a formalin-killed autogenous M bovis bacterin. During the epizootics, the eyes were examined by cultural technique, and isolates which were obtained were categorized by catalase activity, source (diseased or nondiseased eyes), and reactivity with the various sera. The serum reactivity of the isolates was compared with that of the vaccinal strain. The vaccinal strain and 8 of the 1 5 selected isolates obtained during the 1974 epizootic were catalase negative. Seven of the 15 isolates from the 1974 epizootic and all of the selected isolates from the 1975 epizootic were catalase positive. A significantly higher (P less than 0.01) percentage of calf sera were serologically reactive with the vaccinal strain and other catalase-negative isolates (45.0%) than with catalase-positive isolates (34.8%). The results, although not definitive, suggest that there may be antigenic differences among strains of M bovis. These differences should be considered when cattle are vaccinated against IBK under natural conditions of exposure.

  3. Isolation of Moraxella bovis ribosomes and their subsequent use in a vaccine against infectious bovine keratoconjunctivitis.

    Science.gov (United States)

    Pugh, G W; Phillips, M; McDonald, T J; Kopecky, K E

    1981-03-01

    A study was conducted to determine whether a Moraxella bovis ribosomal vaccine would protect calves from infectious bovine keratoconjunctivitis (IBK). Each of 16 calves were given 2 inoculations 21 days apart. Twenty-one days after the 2nd inoculation, 8 of the calves were challenge exposed with a homologous strain culture and 8 calves were challenge exposed to a heterologous strain culture of M bovis. Sedimentation velocity analysis of the ribosomes used in this study indicated that they were mostly 30S and 50S subunits. Chemical assays indicated that the ribosomes were composed of 64% to 65% RNA and 35% to 36% protein. The cesium chloride buoyant density of the ribosomes was 1.62 g/ml. Ribosomes used as antigen gave 1 line of precipitation in a gel-diffusion precipitin test with hyperimmune serum against the whole-cell antigen of the homologous strain of M bovis. The eyes of all the experimentally exposed calves became infected and all calves developed clinical signs of either unilateral or bilateral IBK. None of the sera of the vaccinated calves had detectable precipitins against the ribosomal antigen at the time they were challenge exposed, but most of the sera had precipitins against whole-cell and pilus antigens. The results indicate that M bovis ribosomes, although similar to other bacterial ribosomes, did not protect cattle against IBK.

  4. Molecular and antigenic characterization of a Mycoplasma bovis strain causing an outbreak of infectious keratoconjunctivitis.

    Science.gov (United States)

    Alberti, Alberto; Addis, Maria Filippa; Chessa, Bernardo; Cubeddu, Tiziana; Profiti, Margherita; Rosati, Sergio; Ruiu, Angelo; Pittau, Marco

    2006-01-01

    An unusually high incidence of infectious keratoconjunctivitis followed by pneumonia and arthritis was observed in beef calves of a managed herd. No Moraxella spp. or bacteria other than Mycoplasma spp. were obtained from conjunctival and nasal swabs. A strategy was designed for characterization of bovine mycoplasmas at species and strain level on the basis of a combination of molecular tools and the immunoblotting method. The strategy made it possible to rapidly assign the bacterium responsible for this outbreak to one of the phylogenetic clusters of bovine mycoplasmas delineated in this study and then to identify it as Mycoplasma bovis. The strain, designated Sar 1, showed a 100% 16S rDNA sequence identity with two European strains (120/81 and MC3386) isolated in Germany and Ireland, respectively, and hosts a vsp gene analog to the vspA, vsp422-4, and vsp422-8 genes of the M. bovis reference strain PG45T and of the field strain 422. The use of a cross-reactive rabbit serum developed against the Mycoplasma agalactiae immunodominant antigen P48 confirmed the molecular findings. The immunological response of calves against M. bovis was also investigated. This is the first report on the occurrence of M. bovis on the Island of Sardinia (Italy).

  5. Analysis of Babesia bovis-induced gene expression changes in the cattle tick, Rhipcephalus (Boophilus) microplus.

    Science.gov (United States)

    Boophilus ticks are vectors of Babesia bovis, the protozoan causative agent of cattle fever, a disease which is responsible for significant production losses to cattle producers in much of Africa, Central and South America and Australia. We utilized subtractive cDNA library synthesis techniques to o...

  6. Measuring bovine gamma delta T cell function at the site of Mycobacterium bovis infection

    Science.gov (United States)

    The causative agent of tuberculosis (TB) in cattle is Mycobacterium bovis. The characteristic lesions of bovine TB are well-organized pulmonary granulomas. Gamma delta T cells are a unique subset of nonconventional T cells that play major roles in both the innate and adaptive arms of the immune sys...

  7. Draft genome sequences of Streptococcus bovis strains ATCC 33317 and JB1

    Science.gov (United States)

    We report the draft genome sequences of Streptococcus bovis type strain ATTC 33317 (CVM42251) isolated from cow dung and strain JB1 (CVM42252) isolated from a cow rumen in 1977. Strains were subjected to Next Generation sequencing and the genome sizes are approximately 2 MB and 2.2 MB, respectively....

  8. Analysis of the electromobility of different protein fractions of Bovicola bovis females (Mallophaga: Insecta).

    Science.gov (United States)

    Muñoz Parra, S; Soler Cruz, M D; Benítez Rodríguez, R

    1994-09-01

    The authors characterize in females of Bovicola bovis the total proteins of this species by means of SDS-page determining the R(mb), Rx, Rf and mW of each protein in particular and studying finally the number of protein bands in function with their mW.

  9. The Experimental Study of Safety and Efficacy in Using Bovis Calculus Pharmacopuncture Solution as Eye Drop

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    Hyeongsik Seo

    2009-09-01

    Full Text Available Objectives : This experimental study was performed to investigate the safety and efficacy of Bovis Calculus pharmacopuncture solution manufactured with freezing dryness method to use eye drop. To identify the use of it as eye drop, the eye irritation test of rabbits and the antibacterial test of Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Aspergillus niger, Fusarium oxysporum, and Candida albicans were performed. Methods : 1. The eye irritation test of this material was performed according to the Regulation of Korea Food & Drug Administration(2005. 10. 21, KFDA 2005-60. After Bovis Calculus pharmacopuncture solution was administered in the left eye of the rabbits, eye irritation of the cornea, iris and conjunctiva was observed at 1, 2, 3, 4 & 7day. 2. After administering Bovis Calculus pharmacopuncture solution on bacterial species(Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Aspergillus niger, Fusarium oxysporum, Candida albicans which cause Keratitis, MIC(Minimum Inhibition Concentration and the size of inhibition zone were measured. Anti-bacterial potency was also measured using the size of inhibition zone. Results : 1. After Bovis Calculus pharmacopuncture solution was administered in the left eye of the rabbits, it was found that none of nine rabbits have abnormal signs and weight changes. 2. After Bovis Calculus pharmacopuncture solution was medicated in the left eye of the rabbits, no eye irritation of the cornea, iris and conjunctiva was observed at 1, 2, 3, 4 & 7day. 3. There was no response to MIC on bacterial species (Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Aspergillus niger, Fusarium oxysporum, Candida albicans after Bovis Calculus pharmacopuncture solution was medicated. Conclusions : The present study suggests that Bovis Calculus pharmacopuncture solution is a nontoxic and non-irritant medicine, which does not cause eye irritation in

  10. Mycobacterium bovis infections in slaughter pigs in Mubende district, Uganda: a public health concern

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    Muwonge Adrian

    2012-09-01

    Full Text Available Abstract Background Bovine tuberculosis (TB caused by Mycobacterium bovis is primarily a disease of ruminants, particularly cattle (Bos primigenius and buffalo (Syncerus caffer, and is endemic in most developing countries. To date, studies done in Uganda have documented the prevalence of M. bovis in cattle, humans and wild life, in addition to non-tuberculous mycobacteria in pigs. Pigs are increasingly becoming an important component of the livestock sector and share the human ecosystem in rural Uganda. It is therefore of public health interest that they are not a source of human infections. As a follow up to previously published findings on mycobacteria in pigs, this study was aimed at investigating the occurrence and molecular characteristics of M. bovis detected in slaughter pigs in Mubende district, Uganda. One hundred fifty mesenteric lymph nodes with lesions suggestive of mycobacterial infections were collected from approximately one thousand slaughtered pigs in Mubende district over a period of five months. The isolation and identification of M. bovis was done using conventional mycobacteriological methods. Mycobacteria belonging to the Mycobacterium tuberculosis complex (MTC were identified to species level using deletion analysis. Molecular typing was done using Spoligotyping and MIRU-VNTR analysis. Molecular data were analysed and interpreted using MIRU-VNTR plus, SpolDB4.0 and the Mycobacterium bovis spoligo database. Results Of the examined animals, one boar and two sows from Madudu Sub County were infected with M. bovis which presented as lesions of a deep yellow colour and a grit-like texture in the mesenteric lymph nodes. This represents 2% (3/150 of the lymph nodes where lesions suggestive of mycobacterial infections were detected. Molecular analysis revealed that the isolates from the infected pigs showed identical MIRU-VNTR profile and spoligotype (SB1469. Conclusions This is the first study documenting the occurrence of M

  11. Taxonomy Icon Data: Bacillus subtilis [Taxonomy Icon

    Lifescience Database Archive (English)

    Full Text Available g Bacillus_subtilis_S.png Bacillus_subtilis_NS.png http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Bacillus...+subtilis&t=L http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Bacillus+subtilis&t=NL http://biosciencedbc.jp/taxonomy..._icon/icon.cgi?i=Bacillus+subtilis&t=S http://biosciencedbc.jp/taxonomy..._icon/icon.cgi?i=Bacillus+subtilis&t=NS http://togodb.biosciencedbc.jp/togodb/view/taxonomy_icon_comment_en?species_id=214 ...

  12. Acquired resistance to the 16-membered macrolides tylosin and tilmicosin by Mycoplasma bovis.

    Science.gov (United States)

    Lerner, Uri; Amram, Eytan; Ayling, Roger D; Mikula, Inna; Gerchman, Irena; Harrus, Shimon; Teff, Dina; Yogev, David; Lysnyansky, Inna

    2014-01-31

    The molecular mechanism of acquired resistance to the 16-membered macrolides tylosin (Ty) and tilmicosin (Tm) was investigated in Mycoplasma bovis field isolates. Sequence analysis of domains II and V of the two 23S rRNA alleles and ribosomal proteins L4 and L22 was performed on 54 M. bovis isolates showing different minimal inhibitory concentrations (MIC). The presence of any one of the point mutations G748A, C752T, A2058G, A2059G or A2059C (Escherichia coli numbering) in one or both alleles of the 23S rRNAs was correlated with decreased susceptibility to Ty (8-1024 μg/ml) and to Tm (32 to >256 μg/ml) in 27/27 and 27/31 M. bovis isolates, respectively. Although a single mutation in domain II or V could be sufficient to cause decreased susceptibility to Ty, our data imply that a combination of mutations in two domains is necessary to achieve higher MICs (≥ 128 μg/ml). The influence of a combination of mutations in two domains II and V on enhancement of resistance to Tm was less clear. In addition, the amino acid (aa) substitution L22-Q90H was found in 24/32 representative M. bovis isolates with different MICs, but no correlation with decreased susceptibility to Ty or Tm was identified. Multiple aa substitutions were also identified in the L4 protein, including at positions 185-186 (positions 64 and 65 in E. coli) which are adjacent to the macrolide-binding site. This is the first description of the molecular mechanism of acquired resistance to the 16-membered macrolides in M. bovis.

  13. Cloning and expression of Mycobacterium bovis secreted protein MPB51 in Escherichia coli

    Institute of Scientific and Technical Information of China (English)

    JIANG Xiuyun; WANG Chunfeng; WANG Chunfang; HE Zhaoyang

    2007-01-01

    The purpose of this study is to clone,identify,and express the mature secreted protein MPB51 from Mycobacterium bovis and to lay a good foundation for the diagnosis of M.bovis,for applying M.bovis vaccine into clinical practice,and for detection of immunity effectiveness.The gene encoding MPB51 was amplified from M.bovis Valleel 11 chromosomal DNA by using PCR technique,PCR product was approximately 800 bp DNA segment.Clone vector pGEM-T-51 was successfully constructed by the PCR product that was cloned into pGEM-T vector by using T-A clone technique,pGEM-T-51 and pET28a(+)were digested by BamH I and EcoR I double enzymes.The prokaryotic expression vector pET28a-51 was constructed by using the purified MPB51 gene that was subcloned into the expression vector pET28a(+).Plasmid containing pET28a-51 was transformed into competence E.coli BL21(DE3).The bacterium was induced by IPTG and its lysates were loaded directly onto SDS-PAGE.An approximately 30 kDa exogenous protein was observed on the SDS-PAGE.The protein was analyzed by using Western-blotting and it had the antigenic activity ofM.bovis.These results could serve as a basis for further studies on the usefulness of the gene and its expression product in the development of subunit vaccine and DNA vaccine against bovine tuberculosis.

  14. Diversity of Babesia bovis merozoite surface antigen genes in the Philippines.

    Science.gov (United States)

    Tattiyapong, Muncharee; Sivakumar, Thillaiampalam; Ybanez, Adrian Patalinghug; Ybanez, Rochelle Haidee Daclan; Perez, Zandro Obligado; Guswanto, Azirwan; Igarashi, Ikuo; Yokoyama, Naoaki

    2014-02-01

    Babesia bovis is the causative agent of fatal babesiosis in cattle. In the present study, we investigated the genetic diversity of B. bovis among Philippine cattle, based on the genes that encode merozoite surface antigens (MSAs). Forty-one B. bovis-positive blood DNA samples from cattle were used to amplify the msa-1, msa-2b, and msa-2c genes. In phylogenetic analyses, the msa-1, msa-2b, and msa-2c gene sequences generated from Philippine B. bovis-positive DNA samples were found in six, three, and four different clades, respectively. All of the msa-1 and most of the msa-2b sequences were found in clades that were formed only by Philippine msa sequences in the respective phylograms. While all the msa-1 sequences from the Philippines showed similarity to those formed by Australian msa-1 sequences, the msa-2b sequences showed similarity to either Australian or Mexican msa-2b sequences. In contrast, msa-2c sequences from the Philippines were distributed across all the clades of the phylogram, although one clade was formed exclusively by Philippine msa-2c sequences. Similarities among the deduced amino acid sequences of MSA-1, MSA-2b, and MSA-2c from the Philippines were 62.2-100, 73.1-100, and 67.3-100%, respectively. The present findings demonstrate that B. bovis populations are genetically diverse in the Philippines. This information will provide a good foundation for the future design and implementation of improved immunological preventive methodologies against bovine babesiosis in the Philippines. The study has also generated a set of data that will be useful for futher understanding of the global genetic diversity of this important parasite.

  15. Glutamate Dehydrogenase Is Required by Mycobacterium bovis BCG for Resistance to Cellular Stress.

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    James L Gallant

    Full Text Available We recently reported on our success to generate deletion mutants of the genes encoding glutamate dehydrogenase (GDH and glutamine oxoglutarate aminotransferase (GOGAT in M. bovis BCG, despite their in vitro essentiality in M. tuberculosis. We could use these mutants to delineate the roles of GDH and GOGAT in mycobacterial nitrogen metabolism by using M. bovis BCG as a model for M. tuberculosis specifically. Here, we extended our investigation towards the involvement of GDH and GOGAT in other aspects of M. bovis BCG physiology, including the use of glutamate as a carbon source and resistance to known phagosomal stresses, as well as in survival inside macrophages. We find that gdh is indispensable for the utilization of glutamate as a major carbon source, in low pH environments and when challenged with nitric oxide. On the other hand, the gltBD mutant had increased viability under low pH conditions and was unaffected by a challenge with nitric oxide. Strikingly, GDH was required to sustain M. bovis BCG during infection of both murine RAW 264.7 and bone-marrow derived and macrophages, while GOGAT was not. We conclude that the catabolism of glutamate in slow growing mycobacteria may be a crucial function during infection of macrophage cells and demonstrate a novel requirement for M. bovis BCG GDH in the protection against acidic and nitrosative stress. These results provide strong clues on the role of GDH in intracellular survival of M. tuberculosis, in which the essentiality of the gdh gene complicates knock out studies making the study of the role of this enzyme in pathogenesis difficult.

  16. Endometrial inflammation and abnormal expression of extracellular matrix proteins induced by Mycoplasma bovis in dairy cows.

    Science.gov (United States)

    Guo, Mengyao; Wang, Guoqing; Lv, Tingting; Song, Xiaojing; Wang, Tiancheng; Xie, Guanghong; Cao, Yongguo; Zhang, Naisheng; Cao, Rongfeng

    2014-03-15

    Mycoplasma bovis infection can cause endometrial inflammation leading to infertility and involuntary culling in dairy cows. Because extracellular matrix (ECM) proteins affect the adherence of mycoplasma to eukaryotic cell surface, they may play a role in the pathogenesis of the bacteria. The objective of the present study was to evaluate the endometrial inflammatory response and ECM protein expression induced by M bovis. Endometrial concentrations of inflammatory cytokines tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and mRNA and protein expression of collagen IV (CL-IV), fibronectin (FN), and laminin (LN) were evaluated 10, 20, and 30 days after M bovis intrauterine infusion in breed cows 18 days postpartum. The presence of the bacteria in the uterus was detected by nested polymerase chain reaction and denaturing gradient gel electrophoresis. Endometrial TNF-α, IL-1β, and IL-6 concentrations in the treatment group were greater (P < 0.05) than in the positive and negative control groups 20 and 30 days after infusion. Endometrial CL-IV, FN, and LN mRNA and protein expression increased (P < 0.01) 20 days after infusion in all groups. However, the increase was more pronounced in the treatment group and reactive expressions were greater (P < 0.05) than in the positive and negative control groups 10, 20, and 30 days after infusion. In conclusion, M bovis triggered endometrial inflammatory response and increased CL-IV, FN, and LN mRNA and protein expression. The abnormal expression of ECM these proteins may promote the pathogenic effects of M bovis that lead to endometrial tissue damage and infertility.

  17. Using msa-2b as a molecular marker for genotyping Mexican isolates of Babesia bovis.

    Science.gov (United States)

    Genis, Alma D; Perez, Jocelin; Mosqueda, Juan J; Alvarez, Antonio; Camacho, Minerva; Muñoz, Maria de Lourdes; Rojas, Carmen; Figueroa, Julio V

    2009-12-01

    Variable merozoite surface antigens of Babesia bovis are exposed glycoproteins having a role in erythrocyte invasion. Members of this gene family include msa-1 and msa-2 (msa-2c, msa-2a(1), msa-2a(2) and msa-2b). To determine the sequence variation among B. bovis Mexican isolates using msa-2b as a genetic marker, PCR amplicons corresponding to msa-2b were cloned and plasmids carrying the corresponding inserts were purified and sequenced. Comparative analysis of nucleotide and deduced amino acid sequences revealed distinct degrees of variability and identity among the coding gene sequences obtained from 16 geographically different Mexican B. bovis isolates and a reference strain. Clustal-W multiple alignments of the MSA-2b deduced amino acid sequences performed with the 17 B. bovis Mexican isolates, revealed the identification of three genotypes with a distinct set each of amino acid residues present at the variable region: Genotype I represented by the MO7 strain (in vitro culture-derived from the Mexico isolate) as well as RAD, Chiapas-1, Tabasco and Veracruz-3 isolates; Genotype II, represented by the Jalisco, Mexico and Veracruz-2 isolates; and Genotype III comprising the sequences from most of the isolates studied, Tamaulipas-1, Chiapas-2, Guerrero-1, Nayarit, Quintana Roo, Nuevo Leon, Tamaulipas-2, Yucatan and Guerrero-2. Moreover, these three genotypes could be discriminated against each other by using a PCR-RFLP approach. The results suggest that occurrence of indels within the variable region of msa-2b sequences can be useful markers for identifying a particular genotype present in field populations of B. bovis isolated from infected cattle in Mexico.

  18. Utility of Schistosoma bovis Adult Worm Antigens for Diagnosis of Human Schistosomiasis by Enzyme-Linked Immunosorbent Assay and Electroimmunotransfer Blot Techniques

    Science.gov (United States)

    Pardo, J.; Carranza, C.; Turrientes, M. C.; Arellano, J. L. Pérez; Vélez, R. López; Ramajo, V.; Muro, A.

    2004-01-01

    Immunodiagnostic methods based on the detection of antibodies continue to be the most effective and practical methods for the diagnosis of imported schistosomiasis. Schistosoma bovis is a species whose final natural hosts are bovines, ovines, caprines, and small wild ruminants. Different studies have demonstrated the analogies existing between S. bovis and other Schistosoma species which affect humans. The objective of this work was to evaluate the utility of S. bovis adult worm antigens (AWA) for the diagnosis of imported human schistosomiasis by enzyme-linked immunosorbent assay (ELISA) and electroimmunotransfer blotting (EITB) techniques. By detecting eggs, the ELISA for S. bovis AWA was able to definitively detect imported cases with a sensitivity of 94%. The specificity of the ELISA for S. bovis AWA was 97%. There were no differences between the results of the S. bovis AWA ELISA for patients infected with Schistosoma mansoni and those infected with Schistosoma haematobium. The EITB technique showed bands of 85, 37, and 20 kDa, which are characteristic of infections with Schistosoma spp. Specific bands to indicate infection by different species of Schistosoma have not been detected. The combined use of the ELISA for S. bovis AWA and EITB increased the global sensitivity of the study to 97%. Our findings suggest that the ELISA for S. bovis AWA is a useful test for the immunodiagnosis of imported schistosomiasis and that EITB for detecting S. bovis AWA permits the confirmation of diagnosis when the ELISA for S. bovis AWA is positive. PMID:15539523

  19. Effect of milk fermentation by kefir grains and selected single strains of lactic acid bacteria on the survival of Mycobacterium bovis BCG.

    Science.gov (United States)

    Macuamule, C L S; Wiid, I J; van Helden, P D; Tanner, M; Witthuhn, R C

    2016-01-18

    Mycobacterium bovis that causes Bovine tuberculosis (BTB) can be transmitted to humans thought consumption of raw and raw fermented milk products from diseased animals. Lactic acid bacteria (LAB) used in popular traditional milk products in Africa produce anti-microbial compounds that inhibit some pathogenic and spoilage bacteria. M. bovis BCG is an attenuated non-pathogenic vaccine strain of M. bovis and the aim of the study was to determine the effect of the fermentation process on the survival of M. bovis BCG in milk. M. bovis BCG at concentrations of 6 log CFU/ml was added to products of kefir fermentation. The survival of M. bovis BCG was monitored at 12-h intervals for 72 h by enumerating viable cells on Middlebrook 7H10 agar plates enriched with 2% BD BACTEC PANTA™. M. bovis BCG was increasingly reduced in sterile kefir that was fermented for a period of 24h and longer. In the milk fermented with kefir grains, Lactobacillus paracasei subsp. paracasei or Lactobacillus casei, the viability of M. bovis BCG was reduced by 0.4 logs after 24h and by 2 logs after 48 h of fermentation. No viable M. bovis BCG was detected after 60 h of fermentation. Results from this study show that long term fermentation under certain conditions may have the potential to inactivate M. bovis BCG present in the milk. However, to ensure safety of fermented milk in Africa, fermentation should be combined with other hurdle technologies such as boiling and milk pasteurisation.

  20. Evaluation of Immunogenicity and Protective Efficacy Elicited by Mycobacterium bovis BCG Overexpressing Ag85A Protein against Mycobacterium tuberculosis Aerosol Infection.

    Science.gov (United States)

    Xu, Zheng Zhong; Chen, Xiang; Hu, Ting; Meng, Chuang; Wang, Xiao Bo; Rao, Yan; Zhang, Xiao Ming; Yin, Yue Lan; Pan, Zhi Ming; Jiao, Xin An

    2016-01-01

    Mycobacterium bovis bacillus Calmette-Guérin (BCG) is currently the only vaccine available for preventing tuberculosis (TB), however, BCG has varying success in preventing pulmonary TB. In this study, a recombinant BCG (rBCG::Ag85A) strain overexpressing the immunodominant Ag85A antigen was constructed, and its immunogenicity and protective efficacy were evaluated. Our results indicated that the Ag85A protein was successfully overexpressed in rBCG::Ag85A, and the Ag85A peptide-MHC complexes on draining lymph node dendritic cells of C57BL/6 mice infected with rBCG::Ag85A were detectable 4 h post-infection. The C57BL/6 mice infected with this strain had stronger antigen-specific interferon-gamma (IFN-γ) responses and higher antibody titers than those immunized with BCG, and the protective experiments showed that rBCG::Ag85A can enhance protection against Mycobacterium tuberculosis (M. tuberculosis) H37Rv infection compared to the BCG vaccine alone. Our results demonstrate the potential of rBCG::Ag85A as a candidate vaccine against TB.

  1. Molecular Characterization of Heterologous HIV-1gp120 Gene Expression Disruption in Mycobacterium bovis BCG Host Strain: A Critical Issue for Engineering Mycobacterial Based-Vaccine Vectors

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    Joan Joseph

    2010-01-01

    Full Text Available Mycobacterium bovis Bacillus Calmette-Guérin (BCG as a live vector of recombinant bacterial vaccine is a promising system to be used. In this study, we evaluate the disrupted expression of heterologous HIV-1gp120 gene in BCG Pasteur host strain using replicative vectors pMV261 and pJH222. pJH222 carries a lysine complementing gene in BCG lysine auxotrophs. The HIV-1 gp120 gene expression was regulated by BCG hsp60 promoter (in plasmid pMV261 and Mycobacteria spp. α-antigen promoter (in plasmid pJH222. Among 14 rBCG:HIV-1gp120 (pMV261 colonies screened, 12 showed a partial deletion and two showed a complete deletion. However, deletion was not observed in all 10 rBCG:HIV-1gp120 (pJH222 colonies screened. In this study, we demonstrated that E. coli/Mycobacterial expression vectors bearing a weak promoter and lysine complementing gene in a recombinant lysine auxotroph of BCG could prevent genetic rearrangements and disruption of HIV 1gp120 gene expression, a key issue for engineering Mycobacterial based vaccine vectors.

  2. Hepatitis in Disseminated Bacillus Calmette-Guérin Infection

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    Markus U Göttke

    2000-01-01

    Full Text Available Local immunotherapy with an attenuated live strain of Mycobacterium bovis, bacillus Calmette-Guérin (BCG, is an effective and frequently used treatment for in situ transitional cell carcinoma (TCC of the bladder. Success rates are high, and serious side effects are infrequent but can affect every organ system. A 79-year-old patient with recently diagnosed TCC who was treated with intravesical BCG for a recurrence after initial surgical treatment is reported. After unsuccessful attempts at bladder catheterization with the creation of a false passage for his third treatment, BCG was instilled via a suprapubic catheter the same day and again a week later. Two weeks after the third BCG instillation, the patient presented with profound lethargy and weakness to the point of not being able to get up out of a chair. He was febrile, anorexic, icteric and had hepatosplenomegaly. Disseminated BCG infection was suspected on the basis of history, clinical examination and a liver biopsy that showed noncaseating granulomatous hepatitis. Empirical treatment was started with antituberculous combination therapy. A short course of an oral corticosteroid was given. Clinical improvement was marked and sustained so that the patient could be discharged home for the full six-month course of his treatment. Disseminated BCG infection with granulomatous hepatitis can be severe and life-threatening in cases where a large intravascular inoculum of BCG may have been given inadvertently.

  3. Assessment of an oral Mycobacterium bovis BCG vaccine and an inactivated M. bovis preparation for wild boar in terms of adverse reactions, vaccine strain survival, and uptake by nontarget species

    OpenAIRE

    Beltrán-Beck, Beatriz; Romero, Beatriz; Sevilla, Iker A; Barasona, José A.; Garrido, Joseba M; González-Barrio, David; Díez-Delgado, Iratxe; Minguijón, Esmeralda; Casal, Carmen; Vicente, Joaquín; Gortázar, Christian; Aranaz, Alicia

    2014-01-01

    Wildlife vaccination is increasingly being considered as an option for tuberculosis control. We combined data from laboratory trials and an ongoing field trial to assess the risk of an oral Mycobacterium bovis BCG vaccine and a prototype heat-inactivated Mycobacterium bovis preparation for Eurasian wild boar (Sus scrofa). We studied adverse reactions, BCG survival, BCG excretion, and bait uptake by nontarget species. No adverse reactions were observed after administration of BCG (n = 27) or i...

  4. Experimentally induced infectious bovine keratoconjunctivitis: resistance of vaccinated cattle to homologous and heterologous strains of Moraxella bovis.

    Science.gov (United States)

    Pugh, G W; Hughes, D E; Schulz, V D; Graham, C K

    1976-01-01

    In studies to determine whether vaccination with one strain of Moraxella bovis would protect against challenge with virulent homologous or heterologous strains, calves were intramuscularly inoculated 3 times with formalin-killed M bovis, with 14 days between inoculations. Fourteen days after the 3rd vaccinal dose was given, all calves were exposed to homologous or heterologous virulent cultures of M bovis. The results indicated that vaccination with one strain of M bovis may induce protective immunity against homologous and heterologous challenge exposure; however, because vaccinated cattle resisted infection and disease produced by a homologous strain to a greater extent than they resisted those produced by heterologous strains, polyvalent vaccines or highly immunogenic common antigens may be needed to protect cattle against the numerous strains they might encounter under natural field conditions. There was minimal correlation between the presence of precipitating antibodies against the heterologous strains and the establishment of infection and disease.

  5. Longevity of Mycobacterium bovis in Raw and Traditional Souring Milk as a Function of Storage Temperature and Dose.

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    Anita L Michel

    Full Text Available Unpasteurised fresh and souring dairy products form an essential component of household diets throughout many rural communities in southern Africa. The presence of milk-borne zoonotic pathogens such as Mycobacterium bovis (M. bovis, the causative agent of bovine tuberculosis and zoonotic tuberculosis in humans, constitute a public health threat, especially in remote areas with poor disease surveillance in livestock and highly compromised human health due to HIV/AIDS.In this study we used culture to determine the longevity of M. bovis in experimentally inoculated fresh and naturally souring milk obtained from communal cattle in the KwaZulu-Natal province of South Africa. The effect of bacterial load and storage temperature on the survival of M. bovis was evaluated by spiking mixtures of fresh milk and starter soured milk (aMasi culture with three concentrations of bacteria (10(2, 10(4, 10(7 colony forming units/ml, followed by incubation under controlled laboratory conditions that mimicked ambient indoor (20°C and outdoor (33°C temperatures and periodic sampling and testing over time (0-56 days.M. bovis cultured from samples of the fresh and souring milk was identified by PCR analysis. At the highest spiking concentration (10(7 cfu/ml, M. bovis survived for at least 2 weeks at 20°C; but, at all concentrations in the 33°C treatment, M. bovis was absent by three days after inoculation. Logistic regression analysis was used to assess the effects of bacterial concentration and time since inoculation, as well as determine the potential half-life of M. bovis in raw souring milk. Given the most favourable tested conditions for bacterial survival (20°C, approximately 25% of mycobacteria were alive after one day of storage (95% CI: 9-53%, giving an estimated half-life of M. bovis in raw souring milk of approximately 12 hours (95% CI: 7-27 hours.This study demonstrates that M. bovis may survive in fresh and souring milk for periods of time that

  6. Longevity of Mycobacterium bovis in Raw and Traditional Souring Milk as a Function of Storage Temperature and Dose

    Science.gov (United States)

    Hlokwe, Tiny; Raseleka, Keneilwe; Getz, Wayne M.; Marcotty, Tanguy

    2015-01-01

    Background Unpasteurised fresh and souring dairy products form an essential component of household diets throughout many rural communities in southern Africa. The presence of milk-borne zoonotic pathogens such as Mycobacterium bovis (M. bovis), the causative agent of bovine tuberculosis and zoonotic tuberculosis in humans, constitute a public health threat, especially in remote areas with poor disease surveillance in livestock and highly compromised human health due to HIV/AIDS. Methods In this study we used culture to determine the longevity of M. bovis in experimentally inoculated fresh and naturally souring milk obtained from communal cattle in the KwaZulu-Natal province of South Africa. The effect of bacterial load and storage temperature on the survival of M. bovis was evaluated by spiking mixtures of fresh milk and starter soured milk (aMasi) culture with three concentrations of bacteria (102, 104, 107 colony forming units/ml), followed by incubation under controlled laboratory conditions that mimicked ambient indoor (20°C) and outdoor (33°C) temperatures and periodic sampling and testing over time (0-56 days). Results M. bovis cultured from samples of the fresh and souring milk was identified by PCR analysis. At the highest spiking concentration (107cfu/ml), M. bovis survived for at least 2 weeks at 20°C; but, at all concentrations in the 33°C treatment, M. bovis was absent by three days after inoculation. Logistic regression analysis was used to assess the effects of bacterial concentration and time since inoculation, as well as determine the potential half-life of M. bovis in raw souring milk. Given the most favourable tested conditions for bacterial survival (20°C), approximately 25% of mycobacteria were alive after one day of storage (95% CI: 9-53%), giving an estimated half-life of M. bovis in raw souring milk of approximately 12 hours (95% CI: 7-27 hours). Conclusions This study demonstrates that M. bovis may survive in fresh and souring milk for

  7. Effect of bacille calmette-guerin polysaccharide nucleic acid (BCG-PSN) on the expression of T cell-derived cytokines in patients with atopic dermatitis%卡介菌多糖核酸对特应性皮炎患者外周血T淋巴细胞表达细胞因子的影响

    Institute of Scientific and Technical Information of China (English)

    张丽; 狄正鸿; 马蕾; 陈洪铎; 高兴华

    2010-01-01

    目的 探讨卡介菌多糖核酸治疗前后特应性皮炎(AD)患者外周血表达不同细胞因子的T淋巴细胞频数及其与疾病严重程度的相关性.方法 以双盲、随机、交叉对照方法治疗AD患者,应用流式细胞术检测治疗前后8例AD患者外周血T淋巴细胞内细胞因子IL-4、IL-5、IFN-γ、TNF-α的表达;利用AD皮损面积严重度指数积分(ADASIS)评价AD患者病情严重程度.结果 治疗组IFN-γ+CD8+T淋巴细胞频数差值为(8.06±13.96)%,较之安慰剂组(-6.55±10.49)%明显升高(U=2.26,P<0.05),表达其他细胞因子的T细胞频数在两组间差异无统计学意义(P>0.05).治疗组ADASIS积分下降值为1.56±1.49,高于安慰剂组(-0.05±1.54),U=2.00,P<0.05.结论 卡介菌多糖核酸可能通过纠正T淋巴细胞亚群的免疫失衡状态治疗AD.%Objective To assess the changes in frequency of peripheral T lymphocytes expressing different cytokines in patients with atopic dermatitis (AD) before and after treatment with BCG-PSN and their relationship with disease severity. Methods A randomized, double blinded and placebo cross-over control study was conducted. A total of 8 patients with AD were recruited in this study. Intramuscular BCG-PSN or placebo was given to patients every other day for 36 days. Flow cytometry was performed to measure the frequency of IL4-, IL5-, IFN-γ- and TNFα-expressing peripheral CD4+ T cells and CD8+ T cells before and after the therapy. Disease severity was evaluated by atopic dermatitis area and severity index score (ADASIS). Results The difference value in IFN-γ+CD8+ T cell frequency before and after therapy was significantly higher in patients treated with BCG-PSN than in those with placebo (8.056 ± 13.962 vs -6.549 ± 10.491, U = 2.26, P 0.05). The decrease in ADASIS was 1.56 ± 1.49 in patients treated with BCG-PSN, which was statistically higher than that in placebo-treated patients (-0.05 ± 1.54, U = 2.00, P< 0.05). Conclusion As an immunomodulator, BCG-PSN may control AD by restoring the balance of T-cell subsets.

  8. The prevalence of Mycobacterium bovis-infection and atypical mycobacterioses in cattle in and around Morogoro, Tanzania

    DEFF Research Database (Denmark)

    Durnez, Lies; Sadiki, Harrison; Katakweba, Abdul

    2009-01-01

    -infection and atypical mycobacterioses. Milk samples were taken from tuberculin positive animals and analysed for the presence of mycobacteria. Total prevalences of 2.5% and 10.1% were found for M. bovis-infection and atypical mycobacterioses respectively, with more M. bovis-infection in cattle in the extensive...... herd owners and their families at risk for BTB. Therefore control of BTB, as well as education of cattle owners is important, especially in the extensive sector....

  9. Improved detection of Mycobacterium bovis infection in bovine lymph node tissue using immunomagnetic separation (IMS-based methods.

    Directory of Open Access Journals (Sweden)

    Linda D Stewart

    Full Text Available Immunomagnetic separation (IMS can selectively isolate and concentrate Mycobacterium bovis cells from lymph node tissue to facilitate subsequent detection by PCR (IMS-PCR or culture (IMS-MGIT. This study describes application of these novel IMS-based methods to test for M. bovis in a survey of 280 bovine lymph nodes (206 visibly lesioned (VL, 74 non-visibly lesioned (NVL collected at slaughter as part of the Northern Ireland bovine TB eradication programme. Their performance was evaluated relative to culture. Overall, 174 (62.1% lymph node samples tested positive by culture, 162 (57.8% by IMS-PCR (targeting IS6110, and 191 (68.2% by IMS-MGIT culture. Twelve (6.9% of the 174 culture positive lymph node samples were not detected by either of the IMS-based methods. However, an additional 79 M. bovis positive lymph node samples (27 (13.1% VL and 52 (70.3% NVL were detected by the IMS-based methods and not by culture. When low numbers of viable M. bovis are present in lymph nodes (e.g. in NVLs of skin test reactor cattle decontamination prior to culture may adversely affect viability, leading to false negative culture results. In contrast, IMS specifically captures whole M. bovis cells (live, dead or potentially dormant which are not subject to any deleterious treatment before detection by PCR or MGIT culture. During this study only 2.7% of NVL lymph nodes tested culture positive, whereas 70.3% of the same samples tested M. bovis positive by the IMS-based tests. Results clearly demonstrate that not only are the IMS-based methods more rapid but they have greater detection sensitivity than the culture approach currently used for the detection of M. bovis infection in cattle. Adoption of the IMS-based methods for lymph node testing would have the potential to improve M. bovis detection in clinical samples.

  10. Apoptosis-Inducing Factor Participation in Bovine Macrophage Mycobacterium bovis-Induced Caspase-Independent Cell Death▿

    Science.gov (United States)

    Vega-Manriquez, X.; López-Vidal, Y.; Moran, J.; Adams, L. G.; Gutiérrez-Pabello, J. A.

    2007-01-01

    Mycobacterium tuberculosis complex species survive and replicate in phagosomes of the host cell. Cell death (CD) has been highlighted as one of the probable outcomes in this host-pathogen interaction. Previously, our group demonstrated macrophage apoptosis as a consequence of Mycobacterium bovis infection. In this study, we aimed to identify the contribution of apoptotic effector elements in M. bovis-induced CD. Bovine macrophages were either infected with M. bovis (multiplicity of infection, 10:1) or treated with an M. bovis cell extract (CFE). Structural changes compatible with CD were evaluated. Chromatin condensation was increased three times by the CFE. On the other hand, a terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) assay demonstrated that levels of DNA fragmentation induced by M. bovis and CFE were 53.7% ± 24% and 38.9% ± 14%, respectively, whereas control cells had a basal proportion of 8.9% ± 4.1%. Rates of DNA fragmentation were unaffected by the presence of the pan-caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp (z-VAD). Cells treated with 100 μg of CFE for 12 h had a fivefold decrease in the level of mitochondrial outer membrane permeabilization compared to that of untreated cells. Neither M. bovis infection nor CFE treatment induced activation of caspase 3, 8, or 9. Translocation of apoptosis-inducing factor (AIF) to the nucleus was identified in 32% ± 3.5% and 26.3% ± 4.9% of M. bovis-infected and CFE-treated cells, respectively. Incubation of macrophages with z-VAD prior to infection did not alter the percentage of cells showing AIF translocation. Our data suggest that M. bovis-induced CD in bovine macrophages is caspase independent with AIF participation. PMID:17158896

  11. Utility of a fecal real-time PCR protocol for detection of Mycobacterium bovis infection in African buffalo (Syncerus caffer).

    Science.gov (United States)

    Roug, Annette; Geoghegan, Claire; Wellington, Elizabeth; Miller, Woutrina A; Travis, Emma; Porter, David; Cooper, David; Clifford, Deana L; Mazet, Jonna A K; Parsons, Sven

    2014-01-01

    A real-time PCR protocol for detecting Mycobacterium bovis in feces was evaluated in bovine tuberculosis-infected African buffalo (Syncerus caffer). Fecal samples spiked with 1.42 × 10(3) cells of M. bovis culture/g and Bacille Calmette-Guérin standards with 1.58 × 10(1) genome copies/well were positive by real-time PCR but all field samples were negative.

  12. Genome analysis shows Bacillus axarquiensis is not a later heterotypic synonym of Bacillus mojavensis; reclassification of Bacillus malacitensis and Brevibacterium halotolerans as heterotypic synonyms of Bacillus axarquiensis.

    Science.gov (United States)

    Dunlap, Christopher A; Bowman, Michael J; Schisler, David A; Rooney, Alejandro P

    2016-06-01

    Bacillus axarquiensis and Bacillus malacitensis were previously reported to be later heterotypic synonyms of Bacillus mojavensis, based primarily on DNA-DNA relatedness values. We have sequenced draft genomes of Bacillus axarquiensis NRRL B-41617T and Bacillus malacitensis NRRL B-41618T. Comparative genomics and DNA-DNA relatedness calculations showed that while Bacillus axarquiensis and Bacillus malacitensis are synonymous with each other, they are not synonymous with Bacillus mojavensis. In addition, a draft genome was completed for Brevibacterium halotolerans, a strain long suspected of being a Bacillus subtilis group member based on 16S rRNA similarities (99.8 % with Bacillus mojavensis). Comparative genomics and DNA-DNA relatedness calculations showed that Brevibacterium halotolerans is synonymous with Bacillus axarquiensis and Bacillus malacitensis. The pairwise in silico DNA-DNA hybridization values calculated in comparisons between the three conspecific strains were all greater than 92 %, which is well above the standard species threshold of 70 %. While the pairwise in silico DNA-DNA hybridization values calculated in comparisons of the three conspecific strains with Bacillus mojavensis were all less than 65 %. The combined results of our genotype and phenotype studies showed that Bacillus axarquiensis, Bacillus malacitensis and Brevibacterium halotolerans are conspecific and distinct from Bacillus mojavensis. Because the valid publication of the name Bacillus axarquiensis predates the publication of the name Bacillus malacitensis, we propose that Bacillus malacitensis be reclassified as a synonym of Bacillus axarquiensis. In addition, we propose to reclassify Brevibacterium halotolerans as a synonym of Bacillus axarquiensis. An amended description of Bacillus axarquiensis is provided.

  13. Human Mycobacterium bovis infection in the south-west of Ireland 1983-1992: a comparison with M. tuberculosis.

    LENUS (Irish Health Repository)

    Cotter, T P

    2012-02-03

    Epidemiological and bacteriological aspects of human Mycobacterium bovis disease were investigated in south-west Ireland (counties Cork & Kerry, population 536,000) over the years 1983-92 inclusive and compared to M. tuberculosis. Results showed a small, stable incidence of culture positive M. bovis human disease, mean annual incidence 0.56 per 100,000 population compared to a higher but declining incidence of culture positive M. tuberculosis (15.3 per 100,000 in 1983, 9.0 per 100,000 in 1992). Male patients were the majority, 63.4 per cent of M. bovis; 62.4% of M. tuberculosis (p = 0.03). Fifty three per cent of M. bovis cases (n = 30) were pulmonary, compared to 85% of M. tuberculosis (n = 626; p = 0.0001). M. bovis patients were older (p = 0.02), mean age 58.4 years (SD 18.9) compared to 48.5 (SD 22.2). The mycobacterial smear positive rate was similar in both groups taken as a whole. No rural-urban difference in incidence was found in either disease, suggesting in the case of M. bovis initial infection in childhood via contaminated milk in the pre-pasteurisation era.

  14. Generation of transgenic cattle expressing human β-defensin 3 as an approach to reducing susceptibility to Mycobacterium bovis infection.

    Science.gov (United States)

    Su, Feng; Wang, Yongsheng; Liu, Guanghui; Ru, Kun; Liu, Xin; Yu, Yuan; Liu, Jun; Wu, Yongyan; Quan, Fusheng; Guo, Zekun; Zhang, Yong

    2016-03-01

    Bovine tuberculosis results from infection with Mycobacterium bovis, a member of the Mycobacterium tuberculosis family. Worldwide, M. bovis infections result in economic losses in the livestock industry; cattle production is especially hard-hit by this disease. Generating M. bovis-resistant cattle may potentially mitigate the impact of this disease by reducing M. bovis infections. In this study, we used transgenic somatic cell nuclear transfer to generate cattle expressing the gene encoding human β-defensin 3 (HBD3), which confers resistance to mycobacteria in vitro. We first generated alveolar epithelial cells expressing HBD3 under the control of the bovine MUC1 promoter, and confirmed that these cells secreted HBD3 and possessed anti-mycobacterial capacity. We then generated and identified transgenic cattle by somatic cell nuclear transfer. The cleavage and blastocyst formation rates of genetically modified embryos provided evidence that monoclonal transgenic bovine fetal fibroblast cells have an integral reprogramming ability that is similar to that of normal cells. Five genetically modified cows were generated, and their anti-mycobacterial capacities were evaluated. Alveolar epithelial cells and macrophages from these cattle expressed higher levels of HBD3 protein compared with non-transgenic cells and possessed effective anti-mycobacterial capacity. These results suggest that the overall risk of M. bovis infection in transgenic cattle is efficiently reduced, and support the development of genetically modified animals as an effective tool to reduce M. bovis infection.

  15. Improved specificity for detection of Mycobacterium bovis in fresh tissues using IS6110 real-time PCR

    Directory of Open Access Journals (Sweden)

    Palmer Mitchell V

    2011-08-01

    Full Text Available Abstract Background Culture of M. bovis from diagnostic specimens is the gold standard for bovine tuberculosis diagnostics in the USA. Detection of M. bovis by PCR in tissue homogenates may provide a simple rapid method to complement bacterial culture. A significant impediment to PCR based assays on tissue homogenates is specificity since mycobacteria other than M. bovis may be associated with the tissues. Results Previously published IS6110 based PCR diagnostic assays, along with one developed in house, were tested against environmental mycobacteria commonly isolated from diagnostic tissues submitted to the National Veterinary Services Laboratory. A real-time PCR assay was developed (IS6110_T that had increased specificity over other IS6110 based assays. Of the 13 non-tuberculous mycobacteria tested with IS6110_T only M. wolinskyi was positive. Thirty M. bovis infected tissue homogenates and 18 control tissues were used to evaluate the potential for the assay as a diagnostic test. In this small sample, IS6110_T detected 20/30 samples from M. bovis infected animals and 0/18 control tissues. Conclusions The IS6110_T assay provides a PCR based assay system that is compatible with current diagnostic protocols for the detection of M. bovis in the USA and compliments current testing strategies.

  16. Experimentally induced infections bovine keratoconjunctivitis: effectiveness of a pilus vaccine against exposure to homologous strains of Moraxella bovis.

    Science.gov (United States)

    Pugh, G W; Hughes, D E; Booth, G D

    1977-10-01

    A study was conducted to determine whether vaccination with sonicated pili of Moraxella bovis would protect cattle from subsequent infection and disease when experimentally challenged by exposure to homologous cultures of M bovis. Some calves were intramuscularly inoculated 2 times with pili of M bovis suspended in water, and others were subcutaneously inoculated 2 times with pili of M bovis suspended in oil; 21 days were allowed between inculations. Controls were nonvaccinated calves. Fourteen days after the last inculation, all calves were exposed to virulent homologous cultures of M bovis. The results indicated that vaccination with sonicated pili of M bovis may induce protective immunity against homologous strain challenge exposure. Vaccines in oil that were injected subcutaneously protected to a greater extent than did vaccines in water that were injected intramuscularly. The development of inflammatory nodules at the site of inoculation was associated with resistance to infection and disease. Only 1 of the vaccinated calves that resisted disease lacked precipitating antibodies against sonicated pili at the time of the challenge exposure. This calf had antibodies 2 weeks later.

  17. LIKE IT ACID AND POOR: A STUDY OF ABIOTIC FACTORS INFLUENCING Streptococcus bovis HC5 GROWTH AND BACTERIOCIN PRODUCTION

    Directory of Open Access Journals (Sweden)

    Ana Andréa Teixeira Barbosa

    2015-04-01

    Full Text Available This study aimed to investigate the effect of pH, temperature, growth atmosphere and nutrient availability on bovicin HC5 production by Streptococcus bovis HC5. S. bovis HC5 grew well in complex and basal media under aerobic and anaerobic conditions, but greater bacteriocin yields were recovered from anaerobic cultures. Lactate production and glucose consumption increased if S. bovis HC5 cells were cultivated at pH 7.0 and at 45 ºC, but higher bovicin HC5 activity was recovered from cells grown in acidic conditions and at lower temperatures (39 ºC. Cultures maintained under continuous CO2 flow showed faster growth rates in basal media, but bacteriocin production was always higher if S. bovis was cultivated in anaerobic sealed tubes. These results suggest that acidic pH and anoxic conditions favor bovicin HC5 production by S. bovis HC5. S. bovis HC5 is a unique lactic acid bacterium in its ability to grow and produce high amounts of a potentially useful bacteriocin in simple media. Considering the constrains for bacteriocin production at commercial scale, it appears that bovicin HC5 production could be achieved at lower costs compared to other bacteriocins from lactic acid bacteria.

  18. Effects of Streptococcus bovis Isolated from Bovine Rumen on the Fermentation Characteristics and Nutritive Value of Tanzania Grass Silage.

    Science.gov (United States)

    Zanine, Anderson de Moura; Bonelli, Emerson Alencar; de Souza, Alexandre Lima; Ferreira, Daniele de Jesus; Santos, Edson Mauro; Ribeiro, Marinaldo Divino; Geron, Luiz Juliano Valério; Pinho, Ricardo Martins Araujo

    2016-01-01

    This study aimed to evaluate the effects of Streptococcus bovis on the fermentation characteristics and nutritive value of Tanzania grass silage. Tanzania grass was chopped and left untreated (U) or treated with Streptococcus bovis JB1 at 1 × 10(6) colony-forming units per gram (cfu/g) of fresh forage or Streptococcus bovis HC5 at 1 × 10(6) cfu/g of fresh forage and packed into sixtuplicate laboratory silos. The largest number of enterobacteria, molds and yeast (M&Y) occurred in untreated silages and the smallest populations of enterobacteria and M&Y and the largest numbers of lactic acid bacteria (LAB), at 9.81 and 9.87 log cfu/g, were observed in Streptococcus bovis JB1 and HC5, respectively (P < 0.05). Silages treated with JB1 and HC5 had lower (P < 0.05) silage pHs and concentrations of ammoniacal nitrogen (NH3-N) than untreated silages. The application of Streptococcus bovis JB1 and HC5 resulted in fewer losses through gases and effluents (P < 0.05), which resulted in greater dry matter recovery (DMR) and crude protein recovery (CPR) (P < 0.05). Streptococcus bovis JB1 and HC5 improved the fermentative profile and increased the concentration of crude protein and DMR and CPR in Tanzania grass silage.

  19. Nested PCR Detection and Phylogenetic Analysis of Babesia bovis and Babesia bigemina in Cattle from Peri-Urban Localities in Gauteng Province, South Africa

    OpenAIRE

    2013-01-01

    ABSTRACT Babesia bovis and Babesia bigemina are tick-borne hemoparasites causing babesiosis in cattle worldwide. This study was aimed at providing information about the occurrence and geographical distribution of B. bovis and B. bigemina species in cattle from Gauteng province, South Africa. A total of 268 blood samples collected from apparently healthy animals in 14 different peri-urban localities were tested using previously established nested PCR assays for the detection of B. bovis and B....

  20. Using UHPLC and UV-vis fingerprint method to evaluate substitutes for Swertia mileensis: An endangered medicinal plant

    Directory of Open Access Journals (Sweden)

    Jie Li

    2017-01-01

    Abbreviation used: UHPLC: Ultra high performance liquid chromatography, UV-vis: Ultraviolet-vis, HBV: Anti-hepatitis virus, DNA: Deoxyribonucleic acid, PCA: Principal component analysis, D-GaIN: D-Galactosamine, BCG: Bacille Calmette-Guerin, LPS: Lipopolysaccharide

  1. Native Valve Streptococcus bovis Endocarditis and Refractory Transfusion Dependent Iron Deficiency Anaemia Associated with Concomitant Carcinoma of the Colon: A Case Report and Review of the Literature

    Directory of Open Access Journals (Sweden)

    Abdul Azeez Ahamed Riyaaz

    2016-01-01

    Full Text Available Streptococcus bovis is found as a commensal organism in human gut and may become opportunistically pathogenic. Infective endocarditis is one of the commonest modes of presentation of this infection. The association between Streptococcus bovis endocarditis and colorectal cancer is well recognized. We report a case of Streptococcus bovis endocarditis along with a refractory iron deficiency anaemia associated with concomitant carcinoma of ascending colon in a 63-year-old male. Cooccurrence of these two conditions may cause a challenge in the management. Considering the strong association of colon cancer with Streptococcus bovis endocarditis, a detailed screening colonoscopy is mandatory following the diagnosis of the latter.

  2. Differences in the antimicrobial susceptibility profiles of Moraxella bovis, M. bovoculi and M. ovis

    Directory of Open Access Journals (Sweden)

    Grazieli Maboni

    2015-06-01

    Full Text Available The aim of this study was to determine the differences in the antimicrobial susceptibility profiles of Moraxella bovis, M. bovoculi and M. ovis. Thirty-two strains of Moraxella spp. isolated from cattle and sheep with infectious keratoconjunctivitis were tested via broth microdilution method to determine their susceptibility to ampicillin, cefoperazone, ceftiofur, cloxacillin, enrofloxacin, florfenicol, gentamicin, neomycin, oxytetracycline and penicillin. The results demonstrated that Moraxella spp. strains could be considered sensitive for most of the antimicrobials tested in this study, but differences between the antimicrobial susceptibility profiles of these three Moraxella species were found. M. bovis might differ from other species due to the higher MIC and MBC values it presented.

  3. Iron repressible outer membrane proteins of Moraxella bovis and demonstration of siderophore-like activity.

    Science.gov (United States)

    Fenwick, B; Rider, M; Liang, J; Brightman, A

    1996-02-01

    Moraxella bovis (strain Epp 63), grown in RPMI 1640 medium supplemented with desferrioxamine mesylate (0.05 mg/ml) resulted in cell free culture supernatants with an increased chromeazurol-S response indicating the presence of high affinity iron binding ligand(s). Supernatants of cultures where growth occurred in tryptic soy broth, RPMI 1640, or RPMI 1640-desferrioxamine supplemented with ferrous sulfate (10 micrograms/ml) were negative on the chromeazurol-S test. Growth of M. bovis in RPMI 1640 or RPMI 1640-desferrioxamine medium induced the expression of previously unrecognized outer membrane proteins whose expression was repressed when the medium was supplemented with iron and which were not produced when growth occurred in tryptic soy broth.

  4. Elucidation of the structure of the oligosaccharide from wild type Moraxella bovis Epp63 lipooligosaccharide.

    Science.gov (United States)

    De Castro, Cristina; Grice, I Darren; Daal, Terese-Marie; Peak, Ian R; Molinaro, Antonio; Wilson, Jennifer C

    2014-03-31

    Moraxella bovis is a Gram-negative microorganism that causes Infectious Bovine Keratoconjunctivitis (IBK), colloquially known as 'Pink eye' in cattle worldwide. Lipopolysaccharides/lipooligosaccharides are the predominant glycans on the surface of Gram-negative microorganisms. Structural elucidation of the oligosaccharide structure of the rough phenotype of Moraxella bovis strain Epp63 was determined using GC-MS, methylation analysis, and NMR spectroscopy. The oligosaccharide is a branched structure that comprises 10 sugars in addition to KDO. The unusual features of this oligosaccharide include the fact that the oligosaccharide is devoid of heptose. The KDO residue is directly attached to a (→4,6)-branched glucose and additionally contains a terminal open chain acetal-linked N-acetylgalactosamine, (1S)-GalaNAc residue →4,6-linked to a sub-terminal galactose residue.

  5. Thin layer microcolony culture associated with PCR for early identification of Mycobacterium bovis

    Directory of Open Access Journals (Sweden)

    Tatiana Reis do Rosário

    2014-01-01

    Full Text Available The initial growth of mycobacteria from 49 samples of cattle and buffalo organs collected in commercial slaughterhouses was compared between modified Middlebrook 7H11 thin layer microcolony culture and Stonebrink medium used in the isolation of Mycobacterium bovis. Aliquots were decontaminated by Petroff's method, processed and cultured in both media. The identity of the acid-fast bacilli stained by Ziehl-Neelsen was confirmed by PCR. Optical microscopy showed that results of the early observation of Mycobacterium bovis colonies in thin layer culture were similar to those obtained in macroscopic observation of the colonies in Stonebrink medium. However, early observation of the colonies enabled early confirmation by PCR, given the shorter time to the visualization of colonies when thin layer culture was used (between the 12nd and 25th day of culture.

  6. Analysis of Babesia bovis infection-induced gene expression changes in larvae from the cattle tick, Rhipicephalus (Boophilus microplus

    Directory of Open Access Journals (Sweden)

    Heekin Andrew M

    2012-08-01

    Full Text Available Abstract Background Cattle babesiosis is a tick-borne disease of cattle that has severe economic impact on cattle producers throughout the world’s tropical and subtropical countries. The most severe form of the disease is caused by the apicomplexan, Babesia bovis, and transmitted to cattle through the bite of infected cattle ticks of the genus Rhipicephalus, with the most prevalent species being Rhipicephalus (Boophilus microplus. We studied the reaction of the R. microplus larval transcriptome in response to infection by B. bovis. Methods Total RNA was isolated for both uninfected and Babesia bovis-infected larval samples. Subtracted libraries were prepared by subtracting the B. bovis-infected material with the uninfected material, thus enriching for expressed genes in the B. bovis-infected sample. Expressed sequence tags from the subtracted library were generated, assembled, and sequenced. To complement the subtracted library method, differential transcript expression between samples was also measured using custom high-density microarrays. The microarray probes were fabricated using oligonucleotides derived from the Bmi Gene Index database (Version 2. Array results were verified for three target genes by real-time PCR. Results Ticks were allowed to feed on a B. bovis-infected splenectomized calf and on an uninfected control calf. RNA was purified in duplicate from whole larvae and subtracted cDNA libraries were synthesized from Babesia-infected larval RNA, subtracting with the corresponding uninfected larval RNA. One thousand ESTs were sequenced from the larval library and the transcripts were annotated. We used a R. microplus microarray designed from a R. microplus gene index, BmiGI Version 2, to look for changes in gene expression that were associated with infection of R. microplus larvae. We found 24 transcripts were expressed at a statistically significant higher level in ticks feeding upon a B. bovis-infected calf contrasted to ticks

  7. The Effect of Oral Vaccination with Mycobacterium bovis BCG on the Development of Tuberculosis in Captive European Badgers (Meles meles)

    Science.gov (United States)

    Chambers, Mark A.; Aldwell, Frank; Williams, Gareth A.; Palmer, Si; Gowtage, Sonya; Ashford, Roland; Dalley, Deanna J.; Davé, Dipesh; Weyer, Ute; Salguero, Francisco J.; Nunez, Alejandro; Nadian, Allan K.; Crawshaw, Timothy; Corner, Leigh A. L.; Lesellier, Sandrine

    2017-01-01

    The European badger (Meles meles) is a reservoir host of Mycobacterium bovis and responsible for a proportion of the tuberculosis (TB) cases seen in cattle in the United Kingdom and Republic of Ireland. An injectable preparation of the bacillus Calmette-Guérin (BCG) vaccine is licensed for use in badgers in the UK and its use forms part of the bovine TB eradication plans of England and Wales. However, there are practical limitations to the widespread application of an injectable vaccine for badgers and a research priority is the development of an oral vaccine deliverable to badgers in bait. Previous studies reported the successful vaccination of badgers with oral preparations of 108 colony forming units (CFU) of both Pasteur and Danish strains of BCG contained within a lipid matrix composed of triglycerides of fatty acids. Protection against TB in these studies was expressed as a reduction in the number and apparent progression of visible lesions, and reductions in the bacterial load and dissemination of infection. To reduce the cost of an oral vaccine and reduce the potential for environmental contamination with BCG, it is necessary to define the minimal efficacious dose of oral BCG for badgers. The objectives of the two studies reported here were to compare the efficacy of BCG Danish strain in a lipid matrix with unformulated BCG given orally, and to evaluate the efficacy of BCG Danish in a lipid matrix at a 10-fold lower dose than previously evaluated in badgers. In the first study, both BCG unformulated and in a lipid matrix reduced the number and apparent progression of visible lesions and the dissemination of infection from the lung. In the second study, vaccination with BCG in the lipid matrix at a 10-fold lower dose produced a similar outcome, but with greater intra-group variability than seen with the higher dose in the first study. Further research is needed before we are able to recommend a final dose of BCG for oral vaccination of badgers against TB or

  8. Complete genome sequence of Mycoplasma bovis type strain PG45 (ATCC 25523).

    Science.gov (United States)

    Wise, Kim S; Calcutt, Michael J; Foecking, Mark F; Röske, Kerstin; Madupu, Ramana; Methé, Barbara A

    2011-02-01

    This complete and fully assembled genome sequence of Mycoplasma bovis type strain PG45 is the first available for this species and offers a framework for comparison with additional pathogenic isolates. The single circular chromosome of 1,003,404 bp reveals multiple gene sets and mechanisms involved in variable expression of surface antigens and the incursion of numerous and assorted mobile elements, despite its reduced size.

  9. Complete Genome Sequence of Mycoplasma bovis Type Strain PG45 (ATCC 25523)▿

    Science.gov (United States)

    Wise, Kim S.; Calcutt, Michael J.; Foecking, Mark F.; Röske, Kerstin; Madupu, Ramana; Methé, Barbara A.

    2011-01-01

    This complete and fully assembled genome sequence of Mycoplasma bovis type strain PG45 is the first available for this species and offers a framework for comparison with additional pathogenic isolates. The single circular chromosome of 1,003,404 bp reveals multiple gene sets and mechanisms involved in variable expression of surface antigens and the incursion of numerous and assorted mobile elements, despite its reduced size. PMID:21134966

  10. Spoligotyping and variable number tandem repeat analysis of Mycobacterium bovis isolates from cattle in Brazil

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    Patrícia Martins Parreiras

    2012-02-01

    Full Text Available We performed spoligotyping and 12-mycobacterial interspersed repetitive unit-variable number tandem repeats (MIRU-VNTRs typing to characterise Mycobacterium bovis isolates collected from tissue samples of bovines with lesions suggestive for tuberculosis during slaughter inspection procedures in abattoirs in Brazil. High-quality genotypes were obtained with both procedures for 61 isolates that were obtained from 185 bovine tissue samples and all of these isolates were identified as M. bovis by conventional identification procedures. On the basis of the spoligotyping, 53 isolates were grouped into nine clusters and the remaining eight isolates were unique types, resulting in 17 spoligotypes. The majority of the Brazilian M. bovis isolates displayed spoligotype patterns that have been previously observed in strains isolated from cattle in other countries. MIRU-VNTR typing produced 16 distinct genotypes, with 53 isolates forming eight of the groups, and individual isolates with unique VNTR profiles forming the remaining eight groups. The allelic diversity of each VNTR locus was calculated and only two of the 12-MIRU-VNTR loci presented scores with either a moderate (0.4, MIRU16 or high (0.6, MIRU26 discriminatory index (h. Both typing methods produced similar discriminatory indexes (spoligotyping h = 0.85; MIRU-VNTR h = 0.86 and the combination of the two methods increased the h value to 0.94, resulting in 29 distinct patterns. These results confirm that spoligotyping and VNTR analysis are valuable tools for studying the molecular epidemiology of M. bovis infections in Brazil.

  11. Wrist Tenosynovitis due to Mycobacterium bovis Infection: Case Series and Review of the Literature

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    Mehmet Derviş Güner, MD

    2014-12-01

    Full Text Available Summary: Tuberculosis infections are still one of the most important public health problems among developing countries. Musculoskeletal involvement represents 10–15% of all extrapulmonary cases. Tuberculosis tenosynovitis is usually misdiagnosed as nonspecific tenosynovitis. To avoid misdiagnosis and mistreatment, it is important to be alert for mycobacterial infections. This article presents 3 patients with wrist tenosynovitis, which was caused by Mycobacterium bovis infection. The article also includes review of the literature.

  12. Characterization of pilin genes from seven serologically defined prototype strains of Moraxella bovis.

    OpenAIRE

    1994-01-01

    Numerous field isolates of Moraxella bovis have previously been classified by serological techniques into seven serogroups, each defined by homologous cross-reaction with antisera prepared against purified pili of a single prototype strain. The gene encoding pilin from each of the prototype strains has been characterized by nucleotide sequence determination. The coding sequences show extensive homology (70 to 80%) while the proximal downstream sequences show a dichotomy into nonhomologous set...

  13. Characterisation and quantitation of pilus antigens of Moraxella bovis by ELISA.

    Science.gov (United States)

    Lepper, A W; Hermans, L R

    1986-12-01

    Hyperimmune serums raised in rabbits to purified pili from 9 Australian and 2 American strains of Moraxella bovis from infectious bovine keratoconjunctivitis (IBK) affected herds were used to study the degree of binding between combinations of antigen and antiserum in a conventional enzyme linked immunosorbent assay (ELISA). With the aid of appropriate absorption tests major antigenic differences among pili were found permitting 6 distinct serogroups to be recognised. Further, production of specific antiserums to representative strains of each serogroup in goats facilitated the development of a double antibody sandwich ELISA which could be used to quantitate pilus expression of a given strain of M. bovis, or to differentiate pilus serogroups of 22 strains of M. bovis obtained from a total of 12 Australian herds. Most isolates were found to belong to serogroups designated IV and V. One strain from the United States of America showed total homology with Australian serogroup IV while the other showed some cross-reactivity with serogroups V and VI.

  14. Genotyping Mycobacterium bovis from cattle in the Central Pampas of Argentina: temporal and regional trends

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    Ernesto Shimizu

    2014-04-01

    Full Text Available Mycobacterium bovis is the causative agent of bovine tuberculosis (TB, a disease that affects approximately 5% of Argentinean cattle. Among the molecular methods for genotyping, the most convenient are spoligotyping and variable number of tandem repeats (VNTR. A total of 378 samples from bovines with visible lesions consistent with TB were collected at slaughterhouses in three provinces, yielding 265 M. bovis spoligotyped isolates, which were distributed into 35 spoligotypes. In addition, 197 isolates were also typed by the VNTR method and 54 combined VNTR types were detected. There were 24 clusters and 27 orphan types. When both typing methods were combined, 98 spoligotypes and VNTR types were observed with 27 clusters and 71 orphan types. By performing a meta-analysis with previous spoligotyping results, we identified regional and temporal trends in the population structure of M. bovis. For SB0140, the most predominant spoligotype in Argentina, the prevalence percentage remained high during different periods, varying from 25.5-57.8% (1994-2011. By contrast, the second and third most prevalent spoligotypes exhibited important fluctuations. This study shows that there has been an expansion in ancestral lineages as demonstrated by spoligotyping. However, exact tandem repeat typing suggests dynamic changes in the clonal population of this microorganism.

  15. Comparative genome analysis of Bacillus cereus group genomes with Bacillus subtilis

    OpenAIRE

    Anderson, Iain; Sorokin, Alexei; Kapatral, Vinayak; Reznik, Gary; Bhattacharya, Anamitra; Mikhailova, Natalia; Burd, Henry; Joukov, Victor; Kaznadzey, Denis; Walunas, Theresa; D'Souza, Mark; Larsen, Niels; Pusch, Gordon; Liolios, Konstantinos; Grechkin, Yuri

    2005-01-01

    Genome features of the Bacillus cereus group genomes (representative strains of Bacillus cereus, Bacillus anthracis and Bacillus thuringiensis sub spp israelensis) were analyzed and compared with the Bacillus subtilis genome. A core set of 1,381 protein families among the four Bacillus genomes, with an additional set of 933 families common to the B. cereus group, was identified. Differences in signal transduction pathways, membrane transporters, cell surface structures, cell wall, and S-...

  16. The effect of multiple evolutionary selections on synonymous codon usage of genes in the Mycoplasma bovis genome.

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    Jian-hua Zhou

    Full Text Available Mycoplasma bovis is a major pathogen causing arthritis, respiratory disease and mastitis in cattle. A better understanding of its genetic features and evolution might represent evidences of surviving host environments. In this study, multiple factors influencing synonymous codon usage patterns in M. bovis (three strains' genomes were analyzed. The overall nucleotide content of genes in the M. bovis genome is AT-rich. Although the G and C contents at the third codon position of genes in the leading strand differ from those in the lagging strand (p<0.05, the 59 synonymous codon usage patterns of genes in the leading strand are highly similar to those in the lagging strand. The over-represented codons and the under-represented codons were identified. A comparison of the synonymous codon usage pattern of M. bovis and cattle (susceptible host indicated the independent formation of synonymous codon usage of M. bovis. Principal component analysis revealed that (i strand-specific mutational bias fails to affect the synonymous codon usage pattern in the leading and lagging strands, (ii mutation pressure from nucleotide content plays a role in shaping the overall codon usage, and (iii the major trend of synonymous codon usage has a significant correlation with the gene expression level that is estimated by the codon adaptation index. The plot of the effective number of codons against the G+C content at the third codon position also reveals that mutation pressure undoubtedly contributes to the synonymous codon usage pattern of M. bovis. Additionally, the formation of the overall codon usage is determined by certain evolutionary selections for gene function classification (30S protein, 50S protein, transposase, membrane protein, and lipoprotein and translation elongation region of genes in M. bovis. The information could be helpful in further investigations of evolutionary mechanisms of the Mycoplasma family and heterologous expression of its functionally

  17. Prolactin modulates cytokine production induced by culture filtrate proteins of M. bovis through different signaling mechanisms in THP1 cells.

    Science.gov (United States)

    Martínez-Neri, Priscila A; López-Rincón, Gonzalo; Mancilla-Jiménez, Raúl; del Toro-Arreola, Susana; Muñoz-Valle, José Francisco; Fafutis-Morris, Mary; Bueno-Topete, Miriam Ruth; Estrada-Chávez, Ciro; Pereira-Suárez, Ana Laura

    2015-01-01

    The immunomodulatory functions of prolactin (PRL) are well recognized. Augmented PRL plasma levels were observed in patients with advanced tuberculosis (TB). Recently, we have reported that LPS and Mycobacterium bovis (M. bovis) induced differential expression of PRL receptor (PRLR) isoforms in THP-1 cells and bovine macrophages, respectively. The aim of this work was to determine whether PRL should be considered as a potential modulator of the signaling pathways and cytokine synthesis, induced by culture filtrate protein (CFP) from M. bovis in THP-1 monocytes. The THP-1 cells were stimulated with PRL (20ng/mL), M. bovis CFP (50μg/mL). PRLR as well as phosphorylated STAT3, STAT5, Akt1/2/3, ERK1/2 and p38 expression were evaluated by Western blot. IL1-β, TNF-α, IL-6, IL-12, IL-8, and IL-10 concentrations were measured by ELISA. Our results demonstrated that the expression pattern of PRLR short isoforms is induced by M. bovis CFP. M bovis CFP induced phosphorylation of Akt2, ERK1/2, p38, STAT3, and STAT5 pathways. In turn, PRL only activated the JAK2/STAT3-5 signaling pathway. However, when combined both stimuli, PRL significantly increased STAT3-5 phosphorylation and downregulated Akt2, ERK1/2, and p38 phosphorylation. As expected, M. bovis CFP induced substantial amounts of IL1-β, IL-6, TNF-α, IL-8, IL-12, and IL-10. However, the PRL costimulation considerably decreased IL1-β, TNF-α, and IL-12 secretion, and increased IL-10 production. This results suggest that up-regulation of IL-10 by PRL might be modulating the pro-inflammatory response against mycobacterial antigens through the MAPK pathway.

  18. Differential expression of three members of the multidomain adhesion CCp family in Babesia bigemina, Babesia bovis and Theileria equi.

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    Reginaldo G Bastos

    Full Text Available Members of the CCp protein family have been previously described to be expressed on gametocytes of apicomplexan Plasmodium parasites. Knocking out Plasmodium CCp genes blocks the development of the parasite in the mosquito vector, making the CCp proteins potential targets for the development of a transmission-blocking vaccine. Apicomplexans Babesia bovis and Babesia bigemina are the causative agents of bovine babesiosis, and apicomplexan Theileria equi causes equine piroplasmosis. Bovine babesiosis and equine piroplasmosis are the most economically important parasite diseases that affect worldwide cattle and equine industries, respectively. The recent sequencing of the B. bovis and T. equi genomes has provided the opportunity to identify novel genes involved in parasite biology. Here we characterize three members of the CCp family, named CCp1, CCp2 and CCp3, in B. bigemina, B. bovis and T. equi. Using B. bigemina as an in vitro model, expression of all three CCp genes and proteins was demonstrated in temperature-induced sexual stages. Transcripts for all three CCp genes were found in vivo in blood stages of T. equi, and transcripts for CCp3 were detected in vivo in blood stages of B. bovis. However, no protein expression was detected in T. equi blood stages or B. bovis blood stages or B. bovis tick stages. Collectively, the data demonstrated a differential pattern of expression of three orthologous genes of the multidomain adhesion CCp family by B. bigemina, B. bovis and T. equi. The novel CCp members represent potential targets for innovative approaches to control bovine babesiosis and equine piroplasmosis.

  19. Global distribution of Bartonella infections in domestic bovine and characterization of Bartonella bovis strains using multi-locus sequence typing.

    Science.gov (United States)

    Bai, Ying; Malania, Lile; Alvarez Castillo, Danilo; Moran, David; Boonmar, Sumalee; Chanlun, Aran; Suksawat, Fanan; Maruyama, Soichi; Knobel, Darryn; Kosoy, Michael

    2013-01-01

    Bartonella bovis is commonly detected in cattle. One B. bovis strain was recently isolated from a cow with endocarditis in the USA, suggesting its role as an animal pathogen. In the present study, we investigated bartonella infections in 893 cattle from five countries (Kenya, Thailand, Japan, Georgia, and Guatemala) and 103 water buffaloes from Thailand to compare the prevalence of the infection among different regions and different bovid hosts. We developed a multi-locus sequence typing (MLST) scheme based on nine loci (16S rRNA, gltA, ftsZ, groEL, nuoG, ribC, rpoB, ssrA, and ITS) to compare genetic divergence of B. bovis strains, including 26 representatives from the present study and two previously described reference strains (one from French cows and another from a cow with endocarditis in the USA). Bartonella bacteria were cultured in 6.8% (7/103) of water buffaloes from Thailand; all were B. bovis. The prevalence of bartonella infections in cattle varied tremendously across the investigated regions. In Japan, Kenya, and the Mestia district of Georgia, cattle were free from the infection; in Thailand, Guatemala, and the Dusheti and Marneuli districts of Georgia, cattle were infected with prevalences of 10-90%. The Bartonella isolates from cattle belonged to three species: B. bovis (n=165), B. chomelii (n=9), and B. schoenbuchensis (n=1), with the latter two species found in Georgia only. MLST analysis suggested genetic variations among the 28 analyzed B. bovis strains, which fall into 3 lineages (I, II, and III). Lineages I and II were found in cattle while lineage III was restricted to water buffaloes. The majority of strains (17/28), together with the strain causing endocarditis in a cow in the USA, belonged to lineage I. Further investigations are needed to determine whether B. bovis causes disease in bovids.

  20. Global distribution of Bartonella infections in domestic bovine and characterization of Bartonella bovis strains using multi-locus sequence typing.

    Directory of Open Access Journals (Sweden)

    Ying Bai

    Full Text Available Bartonella bovis is commonly detected in cattle. One B. bovis strain was recently isolated from a cow with endocarditis in the USA, suggesting its role as an animal pathogen. In the present study, we investigated bartonella infections in 893 cattle from five countries (Kenya, Thailand, Japan, Georgia, and Guatemala and 103 water buffaloes from Thailand to compare the prevalence of the infection among different regions and different bovid hosts. We developed a multi-locus sequence typing (MLST scheme based on nine loci (16S rRNA, gltA, ftsZ, groEL, nuoG, ribC, rpoB, ssrA, and ITS to compare genetic divergence of B. bovis strains, including 26 representatives from the present study and two previously described reference strains (one from French cows and another from a cow with endocarditis in the USA. Bartonella bacteria were cultured in 6.8% (7/103 of water buffaloes from Thailand; all were B. bovis. The prevalence of bartonella infections in cattle varied tremendously across the investigated regions. In Japan, Kenya, and the Mestia district of Georgia, cattle were free from the infection; in Thailand, Guatemala, and the Dusheti and Marneuli districts of Georgia, cattle were infected with prevalences of 10-90%. The Bartonella isolates from cattle belonged to three species: B. bovis (n=165, B. chomelii (n=9, and B. schoenbuchensis (n=1, with the latter two species found in Georgia only. MLST analysis suggested genetic variations among the 28 analyzed B. bovis strains, which fall into 3 lineages (I, II, and III. Lineages I and II were found in cattle while lineage III was restricted to water buffaloes. The majority of strains (17/28, together with the strain causing endocarditis in a cow in the USA, belonged to lineage I. Further investigations are needed to determine whether B. bovis causes disease in bovids.

  1. Transmissão transovariana de Babesia bovis em Boophilus microplus: obtenção de cepa de carrapato livre de Babesia spp. Babesia bovis transovarian transmission in Boophilus microplus: obtention of a Babesia free tick strain

    Directory of Open Access Journals (Sweden)

    Tânia Regina Bettin dos Santos

    2000-06-01

    Full Text Available O presente trabalho objetivou o estudo de parte do ciclo da Babesia bovis no seu hospedeiro invertebrado, o carrapato Boophilus microplus. Analisou-se a capacidade de infeccção e transmissão transovariana de B. bovis em partenóginas de B. microplus, alimentadas em bovinos portadores e enfermos por esse protozoário. No 18º dia após a infestação, coletaram-se partenóginas diretamente do corpo dos bovinos e teleóginas após o desprendimento natural, a partir do 21º dia. Todos os grupos foram incubados a 27ºC e umidade relativa superior a 70%. No 5º dia após o início da postura, realizou-se o exame de hemolinfa a fim de diagnosticar a infecção dos ínstares por B. bovis. A ausência de infecção detectada no exame de hemolinfa foi confirmada posteriormente com o teste biológico, revelando que partenóginas não transmitem B. bovis transovarianamente. Esses resultados oferecem uma técnica simplificada para a obtenção de cepas de carrapatos livres de B. bovis.In this experiment part of the life cycle of Babesia bovis in its invertebrate host, the tick Boophilus microplus was studied. In order to evaluate the capacity of infection and transmission of B. bovis were collected semi-engorged females of B. microplus fed on carrier and ill bovines. In the 18th day after infestation, semi-engorged females were collected directly from bovine bodies and after 21st day engorged females dropped on the ground. All the collected groups were incubated at 27°C and relative humidity greater than 70%. At the 5th day of oviposition the diagnosis was made by direct examination of haemolymph smears. The biological test reveled that B. bovis transovarial transmission doesn't happer in semi-engorged females. The results offer a simple techique to obtain strains of ticks free of B. bovis.

  2. Bacillus thuringiensis (Bt)

    Science.gov (United States)

    2004-01-01

    Bacillus thuringiensis (Bt), a natural bacteria found all over the Earth, has a fairly novel way of getting rid of unwanted insects. Bt forms a protein substance (shown on the right) that is not harmful to humans, birds, fish or other vertebrates. When eaten by insect larvae the protein causes a fatal loss of appetite. For over 25 years agricultural chemical companies have relied heavily upon safe Bt pesticides. New space based research promises to give the insecticide a new dimension in effectiveness and applicability. Researchers from the Consortium for Materials Development in Space along with industrial affiliates such as Abott Labs and Pern State University flew Bt on a Space Shuttle mission in the fall of 1996. Researchers expect that the Shuttle's microgravity environment will reveal new information about the protein that will make it more effective against a wider variety of pests.

  3. First data on Eurasian wild boar response to oral immunization with BCG and challenge with a Mycobacterium bovis field strain.

    Science.gov (United States)

    Ballesteros, C; Garrido, J M; Vicente, J; Romero, B; Galindo, R C; Minguijón, E; Villar, M; Martín-Hernando, M P; Sevilla, I; Juste, R; Aranaz, A; de la Fuente, J; Gortázar, C

    2009-11-12

    The Eurasian wild boar (Sus scrofa) is considered a reservoir for bovine tuberculosis (bTB) caused by Mycobacterium bovis and closely related members of the Mycobacterium tuberculosis complex in south-central Spain. The vaccination of wildlife with BCG offers an alternative to culling and to movement restriction for the control of bTB among wildlife reservoirs. In this study, we hypothesized that oral BCG immunization of wild boar would affect the expression of immunoregulatory genes and confer protection against M. bovis. Three groups were used to describe the infection, pathological findings and gene expression profiles in wild boar: BCG-vaccinated and M. bovis-challenged (vaccinated challenged group; N=6), non-vaccinated and M. bovis-challenged (non-vaccinated challenged group; N=4), and non-vaccinated and mock-infected (control group; N=2) animals. M. bovis was isolated from 50% (3/6) and 75% (3/4) of vaccinated challenged and non-vaccinated challenged animals, respectively. All four wild boar from the non-vaccinated challenged group developed bTB-compatible lesions 114 days after challenge. In contrast, only 50% of vaccinated challenged wild boar developed lesions. The PBMC mRNA levels of IL4, RANTES, C3, IFN-gamma and methylmalonyl-CoA mutase (MUT) were analyzed at several days post-vaccination (dpi). When vaccinated challenged animals were compared to controls, all five genes were significantly upregulated at the time of M. bovis infection at 186dpi but IFN-gamma levels were also upregulated at 11 and 46dpi. The C3 and MUT mRNA levels were higher at 46dpi, and 11 and 186dpi, respectively, in vaccinated protected wild boar when compared to non-vaccinated challenged animals. At the end of the experiment (300dpi), the mRNA levels of selected genes were lower in non-vaccinated challenged animals when compared to control wild boar. Exposing wild boar to a dose of 10(4)cfu of M. bovis by the oropharyngeal route is an adequate protocol to produce an infection model

  4. Effects of Streptococcus bovis Isolated from Bovine Rumen on the Fermentation Characteristics and Nutritive Value of Tanzania Grass Silage

    Science.gov (United States)

    Zanine, Anderson de Moura; Bonelli, Emerson Alencar; de Souza, Alexandre Lima; Ferreira, Daniele de Jesus; Santos, Edson Mauro; Ribeiro, Marinaldo Divino; Geron, Luiz Juliano Valério; Pinho, Ricardo Martins Araujo

    2016-01-01

    This study aimed to evaluate the effects of Streptococcus bovis on the fermentation characteristics and nutritive value of Tanzania grass silage. Tanzania grass was chopped and left untreated (U) or treated with Streptococcus bovis JB1 at 1 × 106 colony-forming units per gram (cfu/g) of fresh forage or Streptococcus bovis HC5 at 1 × 106 cfu/g of fresh forage and packed into sixtuplicate laboratory silos. The largest number of enterobacteria, molds and yeast (M&Y) occurred in untreated silages and the smallest populations of enterobacteria and M&Y and the largest numbers of lactic acid bacteria (LAB), at 9.81 and 9.87 log cfu/g, were observed in Streptococcus bovis JB1 and HC5, respectively (P Silages treated with JB1 and HC5 had lower (P silage pHs and concentrations of ammoniacal nitrogen (NH3-N) than untreated silages. The application of Streptococcus bovis JB1 and HC5 resulted in fewer losses through gases and effluents (P silage. PMID:27073806

  5. Universal primers suitable to assess population dynamics reveal apparent mutually exclusive transcription of the Babesia bovis ves1alpha gene.

    Science.gov (United States)

    Zupańska, Agata K; Drummond, Paul B; Swetnam, Daniele M; Al-Khedery, Basima; Allred, David R

    2009-07-01

    Babesia bovis is an intraerythrocytic hemoparasite of widespread distribution, which adversely affects livestock production in many regions of the world. This parasite establishes persistent infections of long duration, at least in part through rapid antigenic variation of the VESA1 protein on the infected-erythrocyte surface. To understand the dynamics of in vivo antigenic variation among the parasite population it is necessary to have sensitive and broadly applicable tools enabling monitoring of variation events in parasite antigen genes. To address this need for B. bovis, "universal" primers for the polymerase chain reaction have been designed for the ves1alpha gene, spanning from exon 2 to near the 3' end of cysteine-lysine-rich domain (CKRD) sequences in exon 3. These primers robustly amplified this segment, with minimal bias, from essentially the entire repertoire of full-length ves1alpha sequences in the B. bovis Mexico isolate genome, and are equivalently present in other isolates. On purified genomic DNA, this primer set can achieve a sensitivity of 10 genome equivalents or less. When applied to the amplification of cDNA derived from the B. bovis C9.1 clonal line evidence consistent with mutually exclusive transcription of the ves1alpha gene was obtained, concomitant with detection of numerous mutational events among members of the parasite population. These characteristics of the primers will facilitate the application of polymerase chain reaction-based methodologies to the study of B. bovis population and antigenic switching dynamics.

  6. The molecular prevalence and MSA-2b gene-based genetic diversity of Babesia bovis in dairy cattle in Thailand.

    Science.gov (United States)

    Simking, Pacharathon; Saengow, Sinsamuth; Bangphoomi, Kunan; Sarataphan, Nachai; Wongnarkpet, Sirichai; Inpankaew, Tawin; Jittapalapong, Sathaporn; Munkhjargal, Tserendorj; Sivakumar, Thillaiampalam; Yokoyama, Naoaki; Igarashi, Ikuo

    2013-11-08

    Bovine babesiosis is an economically significant disease that affects dairy farming operations in Thailand. In the present study, 1824 blood-DNA samples prepared from cattle bred in 4 different regions of the country (North, Northeast, Central, and South) were screened using a nested PCR for the specific detection of Babesia bovis. While the overall prevalence of B. bovis was 8.8%, the Central region of Thailand was found to be a high-risk area of the country, as the prevalence of the parasite was 15.0%. The positive rate was relatively higher among the animals of 1-5 years of age. The genetic diversity among the B. bovis parasites was also studied based on their MSA-2b gene, and the findings showed that the Thai sequences were dispersed across 8 of 13 total clades observed in the phylogram. Three of these clades were formed only of Thai sequences. Similarity among the deduced MSA-2b amino acid sequences determined in the present study was 68.3-100%. In conclusion, the present study found that all the locations surveyed were infected with B. bovis and that the parasite populations in Thailand were genetically diverse. Our findings highlight the need for further studies in Thailand to generate more information before a sound control strategy could be implemented against B. bovis.

  7. Cytokines as effectors and predictors of responses in the treatment of bladder cancer by bacillus Calmette-Guérin.

    Science.gov (United States)

    Liu, Xiaoxuan; Dowell, Alexander C; Patel, Prashant; Viney, Richard P; Foster, Michael C; Porfiri, Emilio; James, Nicholas D; Bryan, Richard T

    2014-06-01

    The most effective intravesical treatment of non-muscle-invasive bladder cancer is instillation of live Mycobacterium bovis bacillus Calmette-Guérin (BCG). BCG stimulates the release of cytokines, contributing directly or indirectly to its effectiveness. However, the function of specific cytokines is not well understood. We have undertaken a nonsystematic review of primary evidence regarding cytokine detection, activation and response in BCG patients. Cytokines IL-2, IL-8 and TNF-α appear to be essential for effective BCG therapy and nonrecurrence, while IL-10 may have an inhibitory effect on BCG responses. IL-2, IL-8, TRAIL and TNF-α are potentially predictive of response to BCG. Alterations in genes encoding cytokines may also affect responses. There are significant data showing the association of certain cytokines with successful BCG treatment, and which may be useful predictive markers. Isolating those cytokines mediating efficacy may hold the key to ameliorating BCG's side effects and improving efficacy and patient compliance.

  8. Analysis of cytokine mRNA expression using a novel chromogenic in situ hybridization method in pulmonary granulomas of cattle experimentally infected by aerosolized Mycobacterium bovis

    Science.gov (United States)

    Mycobacterium bovis is the cause of tuberculosis in most animal species, including cattle and is a serious zoonotic pathogen. In humans, M. bovis infection can result in disease clinically indistinguishable from that caused by Mycobacterium tuberculosis, the cause of most tuberculosis in humans. Reg...

  9. Babesia bovis expresses Bbo-6cys-E, a member of a novel gene family that is homologous to the 6-cys family of Plasmodium

    Science.gov (United States)

    A novel Babesia bovis gene family encoding proteins with similarities to the Plasmodium 6cys protein family was identified by TBLASTN searches of the Babesia bovis genome using the sequence of the P. falciparum PFS230 protein as query, and was termed Bbo-6cys gene family. The Bbo-cys6 gene family co...

  10. A space-time analysis of Mycoplasma bovis: bulk tank milk antibody screening results from all Danish dairy herds in 2013-2014

    DEFF Research Database (Denmark)

    Arede, Margarida; Nielsen, Per Kantsø; Ahmed, Syed Sayeem Uddin;

    2016-01-01

    , using the indirect BIO K 302 M. bovis enzyme-linked immunosorbent assay (ELISA) (Bio-X, Belgium) in bulk tank milk from all dairy herds between June 2013 and July 2014. The objective of this study was to investigate the evolution of the spatial distribution of M. bovis in the Danish dairy herd...

  11. Transfection of babesia bovis by double selection with WR99210 and blasticidin-S and its application for functional analysis of thioredoxin peroxidase-1

    Science.gov (United States)

    Genetic manipulation is an essential technique to analyze gene function; however, limited methods are available for Babesia bovis, a causative pathogen of the globally important cattle disease, bovine babesiosis. To date, two stable transfection systems have been developed for B. bovis, using select...

  12. Infectious endocarditis from Streptococcus bovis associated with colonic carcinoma: case report and literature review Endocardite infecciosa por Streptococcus bovis associada com carcinoma cólico: relato de caso e revisão da literatura

    Directory of Open Access Journals (Sweden)

    Jaques Waisberg

    2002-07-01

    Full Text Available BACKGROUND: Many studies in the literature have warned of the need for investigation of colonic lesions among patients, especially elderly ones, who have bacteremia and/or endocarditis from Streptococcus bovis. Bacteremia and infectious endocarditis from Streptococcus bovis may be related to the presence of neoplastic lesions in the large intestine and hepatic disease. AIM: This report describes a patient who presented infectious endocarditis from Streptococcus bovis associated with colonic carcinoma and tubular-villous adenomas. CONCLUSIONS: The finding of this bacterium among patients with septicemia and/or endocarditis is also related to the presence of villous or tubular-villous adenomas in the large intestine. For this reason, complete and detailed investigation of the large intestine must be performed in patients with infectious endocarditis, even in the absence of intestinal symptoms. An increased incidence of this condition or hepatic dysfunction has been reported among patients with infectious endocarditis from Streptococcus bovis. Patients with infectious endocarditis from Streptococcus bovis and normal colonoscopy may be included in the group at risk for developing colonic cancer. The knowledge that there is an association between endocarditis from Streptococcus bovis and carcinoma of the colon has important clinical implications. If the lesion can be discovered at an early stage, curative resection may become possible.RACIONAL: Diferentes estudos na literatura têm alertado para a necessidade de investigação de lesões cólicas entre doentes, especialmente nos mais idosos, que apresentem bacteremia e/ou endocardite por Streptococcus bovis. A bacteremia e a endocardite infecciosa por S. bovis pode estar relacionada à presença de lesões neoplásicas do intestino grosso e a doença hepática. OBJETIVO: Descrição do caso de doente com endocardite infecciosa por S. bovis associada a carcinoma cólico e adenomas t

  13. Antemortem diagnosis of Mycobacterium bovis infection in free-ranging African lions (Panthera leo) and implications for transmission.

    Science.gov (United States)

    Miller, Michele; Buss, Peter; Hofmeyr, Jennifer; Olea-Popelka, Francisco; Parsons, Sven; van Helden, Paul

    2015-04-01

    Diagnosis of tuberculosis in wildlife often relies on postmortem samples because of logistical challenges and lack of field-friendly techniques for live animal testing. Confirmation of infection through detection of infectious organisms is essential for studying the pathogenesis and epidemiology of disease. We describe the application of a technique to obtain respiratory samples from free-ranging living lions to facilitate detection of viable Mycobacterium bovis under field conditions. We identified M. bovis by mycobacterial culture and PCR in tracheobronchial lavage samples from 8/134 (6.0%) lions tested in Kruger National Park, South Africa. This confirms the respiratory shedding of viable M. bovis in living lions. The implications of these results are that infected lions have the potential to transmit this disease and serve as maintenance hosts.

  14. Assessment of Mycobacterium bovis Deleted in p27-p55 Virulence Operon as Candidate Vaccine against Tuberculosis in Animal Models

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    María V. Bianco

    2014-01-01

    Full Text Available A Mycobacterium bovis knockout in p27-p55 operon was tested as an antituberculosis experimental vaccine in animal models. The mutant MbΔp27-p55 was significantly more attenuated in nude mice than its parental strain but more virulent than BCG Pasteur. Challenge experiments in mice and guinea pigs using M. bovis or M. tuberculosis strains showed similar protection conferred by MbΔp27-p55 mutant than BCG in terms of pathology and bacterial loads in spleen but lower protection than BCG in lungs. When tested in cattle, MbΔp27-p55 did not induce IL-2 expression and induced a very low production of IFNγ, suggesting that the lack of P27/P55 reduces the capacity of M. bovis of triggering an adequate Th1 response.

  15. N-terminal amino acid sequence of Bacillus licheniformis alpha-amylase: comparison with Bacillus amyloliquefaciens and Bacillus subtilis Enzymes.

    OpenAIRE

    Kuhn, H.; Fietzek, P P; Lampen, J O

    1982-01-01

    The thermostable, liquefying alpha-amylase from Bacillus licheniformis was immunologically cross-reactive with the thermolabile, liquefying alpha-amylase from Bacillus amyloliquefaciens. Their N-terminal amino acid sequences showed extensive homology with each other, but not with the saccharifying alpha-amylases of Bacillus subtilis.

  16. Bacillus novalis sp. nov., Bacillus vireti sp. nov., Bacillus soli sp. nov., Bacillus bataviensis sp. nov. and Bacillus drentensis sp. nov., from the Drentse A grasslands.

    Science.gov (United States)

    Heyrman, Jeroen; Vanparys, Bram; Logan, Niall A; Balcaen, An; Rodríguez-Díaz, Marina; Felske, Andreas; De Vos, Paul

    2004-01-01

    A group of 42 isolates were isolated from the soil of several disused hay fields, in the Drentse A agricultural research area (The Netherlands), that were taken out of production at different times. The group represents hitherto-uncultured Bacillus lineages that have previously been found, by a non-cultural method, to be predominant in soil. The strains were subjected to a polyphasic taxonomic study, including (GTG)5-PCR, 16S rDNA sequence analysis, DNA-DNA hybridizations, DNA base-ratio determination, fatty acid analysis and morphological and biochemical characterization. By comparing the groupings obtained by (GTG)5-PCR and 16S rDNA sequence analysis, six clusters of similar strains could be recognized. A DNA-DNA relatedness study showed that these clusters represented five novel genospecies. Further analysis supported the proposal of five novel species in the genus Bacillus, namely Bacillus novalis sp. nov. (type strain IDA3307T=R-15439T=LMG 21837T=DSM 15603T), Bacillus vireti sp. nov. (type strain IDA3632T=R-15447T=LMG 21834T=DSM 15602T), Bacillus soli sp. nov. (type strain IDA0086T=R-16300T=LMG 21838T=DSM 15604T), Bacillus bataviensis sp. nov. (type strain IDA1115T=R-16315T=LMG 21833T=DSM 15601T) and Bacillus drentensis sp. nov. (type strain IDA1967T=R-16337T=LMG 21831T=DSM 15600T).

  17. Ocular immune responses in steers following intranasal vaccination with recombinant Moraxella bovis cytotoxin adjuvanted with polyacrylic acid.

    Science.gov (United States)

    Angelos, John A; Edman, Judy M; Chigerwe, Munashe

    2014-02-01

    Infectious bovine keratoconjunctivitis (IBK) caused by Moraxella bovis is the most common eye disease of cattle. The pathogenesis of M. bovis requires the expression of pili that enable the organism to attach to the ocular surface and an RTX (repeats in the structural toxin) toxin (cytotoxin or hemolysin), which is cytotoxic to corneal epithelial cells. In this pilot study, ocular mucosal immune responses of steers were measured following intranasal (i.n.) vaccination with a recombinant M. bovis cytotoxin adjuvanted with polyacrylic acid. Beef steers were vaccinated with either 500 μg (n = 3) or 200 μg (n = 3) of recombinant M. bovis cytotoxin plus adjuvant. Control group steers (n = 2) were vaccinated with adjuvant alone, and all steers were given a booster on day 21. Antigen-specific tear IgA and tear IgG, tear cytotoxin-neutralizing antibody responses, and serum cytotoxin-neutralizing antibody responses were determined in samples collected prevaccination and on days 14, 28, 42, and 55. Changes in tear antigen-specific IgA levels from day 0 to days 28, 42, and 55 were significantly different between groups; however, in post hoc comparisons between individual group pairs at the tested time points, the differences were not significant. Our results suggest that i.n. vaccination of cattle with recombinant M. bovis cytotoxin adjuvanted with polyacrylic acid effects changes in ocular antigen-specific IgA concentrations. The use of intranasally administered recombinant M. bovis cytotoxin adjuvanted with polyacrylic acid could provide an alternative to parenteral vaccination of cattle for immunoprophylaxis against IBK.

  18. The mycobacterial DNA-binding protein 1 (MDP1 from Mycobacterium bovis BCG influences various growth characteristics

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    Maurischat Sven

    2008-06-01

    Full Text Available Abstract Background Pathogenic mycobacteria such as M. tuberculosis, M. bovis or M. leprae are characterised by their extremely slow growth rate which plays an important role in mycobacterial virulence and eradication of the bacteria. Various limiting factors influence the generation time of mycobacteria, and the mycobacterial DNA-binding protein 1 (MDP1 has also been implicated in growth regulation. Our strategy to investigate the role of MDP1 in mycobacterial growth consisted in the generation and characterisation of a M. bovis BCG derivative expressing a MDP1-antisense gene. Results The expression rate of the MDP1 protein in the recombinant M. bovis BCG containing the MDP1-antisense plasmid was reduced by about 50% compared to the reference strain M. bovis BCG containing the empty vector. In comparison to this reference strain, the recombinant M. bovis BCG grew faster in broth culture and reached higher cell masses in stationary phase. Likewise its intracellular growth in mouse and human macrophages was ameliorated. Bacterial clumping in broth culture was reduced by the antisense plasmid. The antisense plasmid increased the susceptibility of the bacteria towards Ampicillin. 2-D protein gels of bacteria maintained under oxygen-poor conditions demonstrated a reduction in the number and the intensity of many protein spots in the antisense strain compared to the reference strain. Conclusion The MDP1 protein has a major impact on various growth characteristics of M. bovis BCG. It plays an important role in virulence-related traits such as aggregate formation and intracellular multiplication. Its impact on the protein expression in a low-oxygen atmosphere indicates a role in the adaptation to the hypoxic conditions present in the granuloma.

  19. Differences between endocarditis caused by Streptococcus bovis and Enterococcus spp. and their association with colorectal cancer.

    Science.gov (United States)

    Corredoira, J; García-País, M J; Coira, A; Rabuñal, R; García-Garrote, F; Pita, J; Rodríguez-Macías, A; Blanco, M; Lopez-Roses, L; López-Álvarez, M J; Alonso-García, M P

    2015-08-01

    Streptococcus bovis group and Enterococcus spp. share phenotypic characteristics and intestinal habitat. Both have been associated with endocarditis and colorectal neoplasm (CRN). We studied all cases of endocarditis diagnosed between 1988 and 2014 in our centre and caused by S. bovis (109, 48.8 % of the bacteremia) and by Enterococcus spp. (36, 3.4 % of the bacteremia). Patients were seen until death or during a long-term follow-up, in order to rule out a concomitant CRN. The 109 cases of S. bovis endocarditis (SbIE) compared with the 36 caused by enterococci showed: a higher proportion of males (91 % vs. 72 %, p=0.005), more multivalvular involvement (28 % vs. 6 %, p=0.004), embolic complications (44 vs. 22 %, p=0.02) and colorectal neoplasm (64 % vs. 25 %, p=0.001). SbIE showed fewer co-morbidities (32 vs. 58 %, p=0.005), and less frequently urinary infection source (0 vs. 25 %, p=0.001) and healthcare-related infection (2 vs. 44 %, p=0.001). A total of 123 patients were followed up for an extended period (mean: 65.9 ± 57.5 months). During the follow-up, 6 of 28 (21 %) cases with enterococcal endocarditis and 43 of 95 (45.2 %, p=0.01) cases with SbIE developed a new CRN. These neoplasiae appeared a mean of 60.4 months later (range 12-181 months). Among the 43 cases with SbIE and CRN, 12 had had a previously normal colonoscopy and 31 had had a previous CRN and developed a second neoplasm. Cases of SbIE present important differences with those caused by Enterococcus spp. Colonoscopy must be mandatory both in the initial evaluation of SbIE, as during the follow-up period.

  20. Isolation and characterization of Mycobacterium bovis strains from indigenous Zambian cattle using Spacer oligonucleotide typing technique

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    Tryland Morten

    2009-07-01

    Full Text Available Abstract Background Bovine tuberculosis (BTB, caused by Mycobacterium bovis, has remained a major source of concern to public health officials in Zambia. Previous investigations have used traditional epidemiological methods that are unable to identify the causative agent and from which dynamics of disease dispersion is difficult to discern. The objective of this study was to isolate, characterize and determine the genetic diversity and relatedness of M. bovis from major cattle rearing districts in Zambia by spoligotyping. A total of 695 carcasses were examined and 98 tissues had gross post-mortem lesions compatible with BTB. Results Forty-two out of the ninety-eight suspected tissues examined had culture properties characteristic of mycobacteria from which 31 isolates yielded interpretable spoligotypes. This technique showed good discriminatory power (HGDI = 0.98, revealing 10 different spoligotype patterns. Twenty-seven isolates belonged to one cluster with more than 95% similarity and inside the cluster, one predominant spoligotype was found in 20 (64.5% of the isolates tested. The highest number of spoligotypes was observed among samples from Namwala district. Spoligotypes from 26 (83.9% of the isolates belonged to five spoligotypes that have been reported before while the remaining 5 (16.1% isolates had unique spoligotypes that are being reported for the first time; these have been assigned numbers SB1763 to SB1767. Five of the 6 districts had the predominant spoligotype (SB0120. Conclusion The study has described the dispersion patterns of M. bovis in Zambian cattle for the first time and has identified 5 spoligotype patterns specific to Zambia. The observation of an overlap in the spoligotype pattern SB0120 in 5 of the 6 districts suggests the probability of sharing a common source of infection.

  1. High throughput phenotypic analysis of Mycobacterium tuberculosis and Mycobacterium bovis strains' metabolism using biolog phenotype microarrays.

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    Bhagwati Khatri

    Full Text Available Tuberculosis is a major human and animal disease of major importance worldwide. Genetically, the closely related strains within the Mycobacterium tuberculosis complex which cause disease are well-characterized but there is an urgent need better to understand their phenotypes. To search rapidly for metabolic differences, a working method using Biolog Phenotype MicroArray analysis was developed. Of 380 substrates surveyed, 71 permitted tetrazolium dye reduction, the readout over 7 days in the method. By looking for ≥5-fold differences in dye reduction, 12 substrates differentiated M. tuberculosis H37Rv and Mycobacterium bovis AF2122/97. H37Rv and a Beijing strain of M. tuberculosis could also be distinguished in this way, as could field strains of M. bovis; even pairs of strains within one spoligotype could be distinguished by 2 to 3 substrates. Cluster analysis gave three clear groups: H37Rv, Beijing, and all the M. bovis strains. The substrates used agreed well with prior knowledge, though an unexpected finding that AF2122/97 gave greater dye reduction than H37Rv with hexoses was investigated further, in culture flasks, revealing that hexoses and Tween 80 were synergistic for growth and used simultaneously rather than in a diauxic fashion. Potential new substrates for growth media were revealed, too, most promisingly N-acetyl glucosamine. Osmotic and pH arrays divided the mycobacteria into two groups with different salt tolerance, though in contrast to the substrate arrays the groups did not entirely correlate with taxonomic differences. More interestingly, these arrays suggested differences between the amines used by the M. tuberculosis complex and enteric bacteria in acid tolerance, with some hydrophobic amino acids being highly effective. In contrast, γ-aminobutyrate, used in the enteric bacteria, had no effect in the mycobacteria. This study proved principle that Phenotype MicroArrays can be used with slow-growing pathogenic mycobacteria

  2. Molecular cloning and expression of two alpha-amylase genes from Streptococcus bovis 148 in Escherichia coli.

    OpenAIRE

    Satoh, E; Niimura, Y; UCHIMURA, T; Kozaki, M; Komagata, K

    1993-01-01

    The alpha-amylase genes of Streptococcus bovis 148 were cloned in Escherichia coli MC1061, using pBR322. The recombinant plasmids were classified into two groups on the basis of their restriction maps. Southern blot analysis did not show homology between the two types of alpha-amylase genes, and the two alpha-amylase genes existed on the chromosomal DNA of S. bovis 148. The enzymatic properties and N-terminal amino acid sequences of the two purified enzymes produced by the cloned E. coli stra...

  3. Bovine tuberculosis in African buffaloes: observations regarding Mycobacterium bovis shedding into water and exposure to environmental mycobacteria

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    van Helden Paul D

    2007-09-01

    Full Text Available Abstract Background African buffaloes are the maintenance host for Mycobacterium bovis in the endemically infected Kruger National Park (KNP. The infection is primarily spread between buffaloes via the respiratory route, but it is not known whether shedding of M. bovis in nasal and oral excretions may lead to contamination of ground and surface water and facilitate the transmission to other animal species. A study to investigate the possibility of water contamination with M. bovis was conducted in association with a BCG vaccination trial in African buffalo. Groups of vaccinated and nonvaccinated buffaloes were kept together with known infected in-contact buffalo cows to allow natural M. bovis transmission under semi-free ranging conditions. In the absence of horizontal transmission vaccinated and control buffaloes were experimentally challenged with M. bovis. Hence, all study buffaloes in the vaccination trial could be considered potential shedders and provided a suitable setting for investigating questions relating to the tenacity of M. bovis shed in water. Results Serial water samples were collected from the drinking troughs of the buffaloes once per season over an eleven-month period and cultured for presence of mycobacteria. All water samples were found to be negative for M. bovis, but 16 non-tuberculous Mycobacterium spp. isolates were cultured. The non-tuberculous Mycobacterium species were further characterised using 5'-16S rDNA PCR-sequencing, resulting in the identification of M. terrae, M. vaccae (or vanbaalenii, M. engbaekii, M. thermoresistibile as well as at least two species which have not yet been classified. Conclusion The absence of detectable levels of Mycobacterium bovis in the trough water suggests that diseased buffalo do not commonly shed the organism in high quantities in nasal and oral discharges. Surface water may therefore not be likely to play an important role in the transmission of bovine tuberculosis from buffalo

  4. msa-1 and msa-2c gene analysis and common epitopes assessment in Mexican Babesia bovis isolates.

    Science.gov (United States)

    Borgonio, Veronica; Mosqueda, Juan; Genis, Alma D; Falcon, Alfonso; Alvarez, J Antonio; Camacho, Minerva; Figueroa, Julio V

    2008-12-01

    Babesia bovis msa-1 and msa-2c genes belong to the variable merozoite surface antigen gene family. These genes code for antigenic proteins present on the merozoite surface (MSA) and are involved in the parasite invasion to the bovine erythrocyte. Previous studies carried out on MSA-1 have evidenced antigen allelic variation in B. bovis isolates from similar endemic regions, as well as in isolates from different geographic regions of the world (Argentina, Australia, Israel). Studies conducted on MSA-2c, however, have shown that this antigen is widely conserved on isolates from distinct geographic regions. In this study, it was hypothesized that MSA-1 and MSA-2c antigens would contain common epitopes despite the presence of nucleotide sequence differences found in 13 B. bovis isolates and strains collected in geographically distant regions of Mexico. Bioinformatics analysis of the primary structure from DNA fragments derived from PCR amplification, cloning, and sequencing of msa-1 and msa-2c genes from the 13 B. bovis populations revealed that the msa-1 gene product present in the various isolates tested is less conserved among isolates obtained within a similar geographic region in Mexico (51-99.7% sequence identity). Results obtained by immunoblot analysis of B. bovis protein extracts reacted with a monoclonal antibody to MSA-1 42-kDa antigen, conclusively showed cross-reactive common epitopes only in Mexican isolates having high sequence identity (>/=99%, eight isolates). Sequence analysis and multiple alignment of deduced MSA-2c demonstrated a high degree of sequence identity (90-100%) among the Mexican B. bovis isolates and strains. Immunoblot results using a polyclonal antibody to MSA-2c reacted against the protein extracts recognized conserved epitopes in at least nine of the B. bovis isolates. The results obtained in this study agree with those previously reported by other researchers and confirm that, based in sequence identity conservation in Mexican B

  5. Immune response to Mycobacterium bovis-an5 infection in genetically selected mice (selection IV-A)

    OpenAIRE

    2007-01-01

    Mice genetically selected for high (H) and low (L) antibody production (Selection IV-A) were used as murine experimental model. The aim of the present work was to evaluate the macrophagic activity and to characterize the immune response in Mycobacterium bovis-AN5 infected mice (3X10(7) bacteria). The response profile previously observed in such strains was not similar to that obtained during M. bovis infection; however, it corroborated works carried out using Selection I, which is very simila...

  6. Análisis transcripcional de la región genética RvD1 de Mycobacterium bovis Transcriptional analysis of genetic region RvD1 of Mycobacterium bovis

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    del Portillo Patricia

    2004-12-01

    Full Text Available Mycobacterium bovis comparte una identidad del 99,9% con los genomas de M. tuberculosis, M. africanum y M. microti. Dentro del 0,1% de esta diferencia se encuentran dos regiones genéticas propias de M. bovis: RvD1 y RvD2, las cuales se encuentran delecionadas del genoma de M. tuberculosis H37Rv y, según el análisis bioin-formático, contienen probables marcos abiertos de lectura (Open Reading Frames: ORF. Con el fin de deter­minar si la región RvD1, transcribe los ORF predichos por bioinformática: ORF1, ORF2 y Rv2024, se extrajeron muestras de ARN total de M. bovis BCG Pasteur, en diferentes puntos de una curva de crecimiento micobacteriano, las cuales fueron analizadas mediante la técnica de Transcripción Reversa y Reacción en Ca­dena de la Polimerasa (RTq-PCR en tiempo real. Los hallazgos obtenidos en esta cinética de transcripción por RTq-PCR en tiempo real demostraron que los probables marcos de lectura abiertos ORF1, ORF2 y Rv2024 de la región RvD1 de M. bovis, sí se transcriben y lo hacen de manera constitutiva, hecho que no había sido repor­tado. Los resultados de esta investigación sirven como un primer paso para determinar la función que desem­peña la región RvD1 de M. bovis, y su posible papel en la patogénesis y en la interacción huésped-patógeno de la tuberculosis bovina y humana. Palabras clave: Mycobacterium bovis, BCG, RNA, RT-PCR, RvD1.Mycobacterium bovis, shares 99.9% of genomic identity with M. tuberculosis, M. africanum and M. microti. Within this 0.1 % of difference, there are two genetic regions characteristics of M. bovis that are deleted in M. tuberculo­sis H37Rv: RvD1 and RvD2. According to bioinformatic analysis, these regions contain Open Reading Frames (ORFs. With the purpose of determining if the RvD1 region transcribes the ORFs predicted by bioinformatics (ORF1, ORF2 and Rv2024; total RNA was extracted from a culture of M. bovis BCG Pasteur, at different time points along the growth curve

  7. Prevalencia e incidencia de Babesia bovis y Babesia bigemina en un hato bovino en Axochiapan, Morelos

    OpenAIRE

    Rojas, E; Domínguez, P; M. García; C. Cruz-Vázquez; Figueroa, J.; Ramos, J

    2004-01-01

    Se describe la prevalencia e incidencia de B. bovis y B. bigemina, en un hato de Axochiapan, Morelos (México), en un periodo de seis meses. El hato estuvo formado por 60 animales encastados de Cebú con Suizo Americano, formando dos grupos, 1 pero < 5 años (2). Se tomaron muestras de sangre y suero cada 30 días, de enero a agosto de 1997, realizando frotis sanguíneo teñido con GIEMSA, determinación del volumen celular aglomerado (VCA) y la prueba de Inmunofluorescencia ...

  8. Molecular characterization of a new Babesia bovis thrombospondin-related anonymous protein (BbTRAP2.

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    Mohamad Alaa Terkawi

    Full Text Available A gene encoding a Babesia bovis protein that shares significant degree of similarity to other apicomplexan thrombospondin-related anonymous proteins (TRAPs was found in the genomic database and designated as BbTRAP2. Recombinant protein containing a conserved region of BbTRAP2 was produced in E. coli. A high antigenicity of recombinant BbTRAP2 (rBbTRAP2 was observed with field B. bovis-infected bovine sera collected from geographically different regions of the world. Moreover, antiserum against rBbTRAP2 specifically reacted with the authentic protein by Western blot analysis and an indirect fluorescent antibody test. Three bands corresponding to 104-, 76-, and 44-kDa proteins were identified in the parasite lysates and two bands of 76- and 44-kDa proteins were detected in the supernatant of cultivated parasites, indicating that BbTRAP2 was proteolytically processed and shed into the culture. Apical and surface localizations of BbTRAP2 were observed in the intracellular and extracellular parasites, respectively, by confocal laser microscopic examination. Moreover, native BbTRAP2 was precipitated by bovine erythrocytes, suggesting its role in the attachment to erythrocytes. Furthermore, the specific antibody to rBbTRAP2 inhibited the growth of B. bovis in a concentration-dependent manner. Consistently, pre-incubation of the free merozoites with the antibody to rBbTRAP2 resulted in an inhibition of the parasite invasion into host erythrocytes. Interestingly, the antibody to rBbTRAP2 was the most inhibitive for the parasite's growth as compared to those of a set of antisera produced against different recombinant proteins, including merozoite surface antigen 2c (BbMSA-2c, rhoptry-associated protein 1 C-terminal (BbRAP-1CT, and spherical body protein 1 (BbSBP-1. These results suggest that BbTRAP2 might be a potential candidate for development of a subunit vaccine against B. bovis infection.

  9. A Complication of BCG Vaccine: A Case of Localized Cutaneous Abscess due to Mycobacterium bovis

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    Nathalie Lussier

    1999-01-01

    Full Text Available The attenuated bacille Calmette-Guérin (BCG vaccine is administered to prevent tuberculosis. Complications of vaccination are uncommon. A case of cutaneous abscess due to BCG is presented in a 24-year-old woman. The abscess developed at the inoculation site four weeks after vaccination. Routine Gram stain and bacterial cultures of the pus were negative. The auramine stain was positive. Mycobacterial cultures were positive after 14 and 18 days, using the BACTEC 12B bottle and Löwenstein-Jensen media, respectively. The mycobacteria were identified as Mycobacterium bovis, vaccinal strain by high-performance liquid chromatography and DNA probe assays.

  10. Uso de tuberculina en caninos para buscar reservorios de Mycobacterium bovis

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    Carlos Bernal-Suarez

    2013-07-01

    Full Text Available Objetivo. Utilizar la prueba de tuberculina en caninos para buscar reservorios de Mycobacterium bovis. Materiales y métodos. Se utilizó la prueba comparada en los caninos que dieron positivo a la PPD ano-caudal, y la técnica de Ziehl-Neelsen en los animales dudosos, para corroborar el resultado. Resultados. Los caninos no son reservorios de la enfermedad, por tal razón, se sugiere utilizar pruebas diagnósticas más específicas para identificar el Mycobacterium en ellos.

  11. Protective and therapeutic efficacy of Mycobacterium smegmatis expressing HBHA-hIL12 fusion protein against Mycobacterium tuberculosis in mice.

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    Shanmin Zhao

    Full Text Available Tuberculosis (TB remains a major worldwide health problem. The only vaccine against TB, Mycobacterium bovis Bacille Calmette-Guerin (BCG, has demonstrated relatively low efficacy and does not provide satisfactory protection against the disease. More efficient vaccines and improved therapies are urgently needed to decrease the worldwide spread and burden of TB, and use of a viable, metabolizing mycobacteria vaccine may be a promising strategy against the disease. Here, we constructed a recombinant Mycobacterium smegmatis (rMS strain expressing a fusion protein of heparin-binding hemagglutinin (HBHA and human interleukin 12 (hIL-12. Immune responses induced by the rMS in mice and protection against Mycobacterium tuberculosis (MTB were investigated. Administration of this novel rMS enhanced Th1-type cellular responses (IFN-γ and IL-2 in mice and reduced bacterial burden in lungs as well as that achieved by BCG vaccination. Meanwhile, the bacteria load in M. tuberculosis infected mice treated with the rMS vaccine also was significantly reduced. In conclusion, the rMS strain expressing the HBHA and human IL-12 fusion protein enhanced immunogencity by improving the Th1-type response against TB, and the protective effect was equivalent to that of the conventional BCG vaccine in mice. Furthermore, it could decrease bacterial load and alleviate histopathological damage in lungs of M. tuberculosis infected mice.

  12. Nanoparticulation of BCG-CWS for application to bladder cancer therapy.

    Science.gov (United States)

    Nakamura, Takashi; Fukiage, Masafumi; Higuchi, Megumi; Nakaya, Akihiro; Yano, Ikuya; Miyazaki, Jun; Nishiyama, Hiroyuki; Akaza, Hideyuki; Ito, Toshihiro; Hosokawa, Hiroyuki; Nakayama, Toshinori; Harashima, Hideyoshi

    2014-02-28

    The Mycobacterium bovis Bacille Calmette-Guerin cell wall skeleton (BCG-CWS) could be used to replace live BCG as a bladder cancer drug. However, because BCG-CWS is poorly soluble, has a strong-negative charge, very high molecular weight and heterogeneity in size of tens of μm, it cannot be used in such an application. We report herein on the development of a novel packaging method that permits BCG-CWS to be encapsulated into 166nm-sized lipid particles. The BCG-CWS encapsulated nano particle (CWS-NP) has a high uniformity and can be easily dispersed. Thus, it has the potential for use as a packaging method that would advance the scope of applications of BCG-CWS as a bladder cancer drug. In a functional evaluation, CWS-NP was efficiently taken up by mouse bladder tumor (MBT-2) cells in vitro and inhibited tumor growth in mice bearing MBT-2 tumors. Moreover, intravesically administered CWS-NP showed significant antitumor effects in a rat model with naturally developed bladder cancer. An enhancement in Th1 differentiation by CWS-NP was also confirmed in human T cells. In conclusion, CWS-NP represents a promising delivery system for BCG-CWS for clinical development as a potent bladder cancer drug.

  13. Screening and Assessing 11 Mycobacterium tuberculosis Proteins as Potential Serodiagnostical Markers for Discriminating TB Patients from BCG Vaccinees

    Institute of Scientific and Technical Information of China (English)

    Guoqiang Zhang; Lingxia Zhang; Mingcheng Zhang; Linlin Pan; Fengyu Wang; Jun Huang; Guoli Li; Jun Yu; Songnian Hu

    2009-01-01

    Purified protein derivative(PPD)skin tests often yield poor specificity, so that to develop new serological antigens for distinguishing between Mycobacterium tu-berculosis infection and Bacille Calmette-Guerin(BCG)vaccination is a priority, especially for developing countries like China. We predicted the antigenicity for selected open reading frames(ORFs)based on the genome sequences of M. tu-berculosis H37Rv and M. bovis BCG, as well as their functions and differences of expression under different stimulus. The candidate ORFs were cloned from H37Rv sequences and expressed as recombinant proteins in Escherichia coll. We studied the serodiagnostic potential of 11 purified recombinants by using enzyme-linked immunosorbent assay(ELISA)and involving a cohort composed of 58 TB patients (34 males and 24 females), 8 healthy volunteers and 50 PPD-negative individuals before and after BCG vaccination. For all the 11 antigens, the median OD val-ues for the sera from TB patients were statistically significantly higher than those for PPD-negative individuals before or after BCG vaccination(P<0.01). They had at least 92% specificity in healthy controls and six seroantigens(Rv0251c, Rv1973, Rv2376c, Rv2537c, Rv2785c and Rv3873A)were never reported with seroantigenicities previously. Thus the approach combining comparative genomies, bioinformatics and ELISA techniques can be employed to identify new seroantigens distinguishing M. tuberculosis infection from BCG vaccination.

  14. An Evaluation of Quantitative PCR Assays (TaqMan® and SYBR Green for the Detection of Babesia bigemina and Babesia bovis, and a Novel Fluorescent-ITS1-PCR Capillary Electrophoresis Method for Genotyping B. bovis Isolates

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    Bing Zhang

    2016-09-01

    Full Text Available Babesia spp. are tick-transmitted haemoparasites causing tick fever in cattle. In Australia, economic losses to the cattle industry from tick fever are estimated at AUD$26 Million per annum. If animals recover from these infections, they become immune carriers. Here we describe a novel multiplex TaqMan qPCR targeting cytochrome b genes for the identification of Babesia spp. The assay shows high sensitivity, specificity and reproducibility, and allows quantification of parasite DNA from Babesia bovis and B. bigemina compared to standard PCR assays. A previously published cytochrome b SYBR Green qPCR was also tested in this study, showing slightly higher sensitivity than the Taqman qPCRs but requires melting curve analysis post-PCR to confirm specificity. The SYBR Green assays were further evaluated using both diagnostic submissions and vaccinated cattle (at 7, 9, 11 and 14 days post-inoculation showed that B. bigemina can be detected more frequently than B. bovis. Due to fewer circulating parasites, B. bovis detection in carrier animals requires higher DNA input. Preliminary data for a novel fluorescent PCR genotyping based on the Internal Transcribed Spacer 1 region to detect vaccine and field alleles of B. bovis are described. This assay is capable of detecting vaccine and novel field isolate alleles in a single sample.

  15. Phages Preying on Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis: Past, Present and Future

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    Annika Gillis

    2014-07-01

    Full Text Available Many bacteriophages (phages have been widely studied due to their major role in virulence evolution of bacterial pathogens. However, less attention has been paid to phages preying on bacteria from the Bacillus cereus group and their contribution to the bacterial genetic pool has been disregarded. Therefore, this review brings together the main information for the B. cereus group phages, from their discovery to their modern biotechnological applications. A special focus is given to phages infecting Bacillus anthracis, B. cereus and Bacillus thuringiensis. These phages belong to the Myoviridae, Siphoviridae, Podoviridae and Tectiviridae families. For the sake of clarity, several phage categories have been made according to significant characteristics such as lifestyles and lysogenic states. The main categories comprise the transducing phages, phages with a chromosomal or plasmidial prophage state, γ-like phages and jumbo-phages. The current genomic characterization of some of these phages is also addressed throughout this work and some promising applications are discussed here.

  16. [Simultaneous determination of seven residual solvents in bovis calculus artifactus by headspace gas chromatography].

    Science.gov (United States)

    Chi, Shuyao; Wu, Dike; Sun, Jinhong; Ye, Ruhan; Wang, Xiaoyan

    2014-05-01

    A headspace gas chromatography (HS-GC) method was developed for the simultaneous determination of seven residual solvents (petroleum ether (60-90 degrees C), acetone, ethyl acetate, methanol, methylene chloride, ethanol and butyl acetate) in bovis calculus artifactus. The DB-WAX capillary column and flame ionization detector (FID) were used for the separation and detection of the residual solvents, and the internal standard method was used for the quantification. The chromatographic conditions, such as equilibrium temperature and equilibrium time, were optimized. Under the optimized conditions, all of the seven residual solvents showed good linear relationships with good correlation coefficients (not less than 0.999 3) in the prescribed concentration range. At three spiked levels, the recoveries for the seven residual solvents were 94.7%-105.2% with the relative standard deviations (RSDs) less than 3.5%. The limits of detection (LODs) of the method were 0.43-5.23 mg/L, and the limits of quantification (LOQs) were 1.25-16.67 mg/L. The method is simple, rapid, sensitive and accurate, and is suitable for the simultaneous determination of the seven residual solvents in bovis calculus artifactus.

  17. The Effect of Mycobacterium avium Complex Infections on Routine Mycobacterium bovis Diagnostic Tests

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    Claire Barry

    2011-01-01

    Full Text Available Bovine tuberculosis (bTB is diagnosed in naturally infected populations exposed to a wide variety of other pathogens. This study describes the cell-mediated immune responses of cattle exposed to Mycobacterium avium subspecies paratuberculosis (Map and Mycobacterium avium subspecies avium with particular reference to routine antefmortem Mycobacterium bovis diagnostic tests. The IFN-γ released in response to stimulated blood was found to peak later in the Map-exposed group and was more sustained when compared to the Maa-exposed group. There was a very close correlation between the responses to the purified protein derivatives (PPD used for stimulation (PPDa, PPDb, and PPDj with PPDa and PPDj most closely correlated. On occasion, in the Map-infected cattle, PPDb-biased responses were seen compared to PPDa suggesting that some Map-infected cattle could be misclassified as M. bovis infected using this test with these reagents. This bias was not seen when PPDj was used. SICCT results were consistent with the respective infections and all calves would have been classed skin test negative.

  18. Pathogenicity and immunogenicity of piliated and nonpiliated phases of Moraxella bovis in calves.

    Science.gov (United States)

    Jayappa, H G; Lehr, C

    1986-10-01

    Pathogenicity evaluations of Moraxella bovis strain EPP 63 grown in the piliated and nonpiliated phase indicated that the organism grown in the piliated phase induced clinical keratoconjunctivitis, whereas the organism grown in the nonpiliated phase did not induce disease under identical challenge conditions. Calves inoculated with culture grown in the piliated phase developed significantly (P less than 0.01) higher lesion scores than did calves inoculated with culture grown in the nonpiliated phase. Vaccination/challenge exposure evaluations indicated that calves immunized with vaccine prepared from piliated culture had significantly lower lesion scores than did control calves (P less than 0.001) or calves immunized with vaccine prepared from nonpiliated culture (P less than 0.05). Similarly, calves immunized with piliated vaccine developed a significantly higher antibody titer against purified pili. Nonvaccinated controls and calves immunized with nonpiliated vaccine did not develop a significant increase in antibody titer. The results indicate that M bovis pili constitute an important factor in the pathogenicity of keratoconjunctivitis and may be used as a vaccine in the control of keratoconjunctivitis in cattle.

  19. Reproduction of bovine keratoconjunctivitis with a purified haemolytic and cytotoxic fraction of Moraxella bovis.

    Science.gov (United States)

    Beard, M K; Moore, L J

    1994-09-01

    Evidence that the beta-haemolysin produced in vitro by Moraxella bovis is an important virulence determinant in vivo was strengthened by studies using a haemolytic preparation of greater purity than previously available. A concentrated haemolytic fraction containing outer-membrane bound vesicles was separated from the cell-free filtrate of a bacterial culture using a process comprising tangential flow ultrafiltration, ion-exchange and gel-filtration high-performance liquid chromatography and centrifugal-driven filtration. The cytotoxicity of haemolytic fractions for calf-corneal epithelial cells in vitro was investigated at progressive stages of this attempted haemolysin purification procedure and the results demonstrated a positive correlation for the levels of haemolytic and cytotoxic activity throughout. Further support for the role of the haemolysin was obtained in vivo following the intra-corneal injection of calves with a crude or a purified haemolytic fraction. The ocular damage caused by both preparations, together with the healing processes and microscopic pathology of the experimentally induced damage closely resembled published descriptions of naturally occurring infectious bovine keratoconjunctivitis. No effect was obtained in vitro or in vivo from equivalent fractions prepared from a non-haemolytic strain of M. bovis.

  20. New Recombinant Mycobacterium bovis BCG Expression Vectors: Improving Genetic Control over Mycobacterial Promoters

    Science.gov (United States)

    Kanno, Alex I.; Goulart, Cibelly; Rofatto, Henrique K.; Oliveira, Sergio C.; Leite, Luciana C. C.

    2016-01-01

    The expression of many antigens, stimulatory molecules, or even metabolic pathways in mycobacteria such as Mycobacterium bovis BCG or M. smegmatis was made possible through the development of shuttle vectors, and several recombinant vaccines have been constructed. However, gene expression in any of these systems relied mostly on the selection of natural promoters expected to provide the required level of expression by trial and error. To establish a systematic selection of promoters with a range of strengths, we generated a library of mutagenized promoters through error-prone PCR of the strong PL5 promoter, originally from mycobacteriophage L5. These promoters were cloned upstream of the enhanced green fluorescent protein reporter gene, and recombinant M. smegmatis bacteria exhibiting a wide range of fluorescence levels were identified. A set of promoters was selected and identified as having high (pJK-F8), intermediate (pJK-B7, pJK-E6, pJK-D6), or low (pJK-C1) promoter strengths in both M. smegmatis and M. bovis BCG. The sequencing of the promoter region demonstrated that it was extensively modified (6 to 11%) in all of the plasmids selected. To test the functionality of the system, two different expression vectors were demonstrated to allow corresponding expression levels of the Schistosoma mansoni antigen Sm29 in BCG. The approach used here can be used to adjust expression levels for synthetic and/or systems biology studies or for vaccine development to maximize the immune response. PMID:26850295

  1. Mycobacterium bovis infection of cattle and white-tailed deer: Translational research of relevance to human tuberculosis

    Science.gov (United States)

    Tuberculosis (TB) is a premier example of a disease complex with pathogens primarily affecting humans (i.e., Mycobacterium tuberculosis) or livestock and wildlife (i.e., Mycobacterium bovis) and with a long history of inclusive collaborations between physicians and veterinarians. Advances with the s...

  2. Molecular detection of Ehrlichia canis, Anaplasma bovis, Anaplasma platys, Candidatus Midichloria mitochondrii and Babesia canis vogeli in ticks from Israel.

    Science.gov (United States)

    Harrus, S; Perlman-Avrahami, A; Mumcuoglu, K Y; Morick, D; Eyal, O; Baneth, G

    2011-03-01

    : Ticks are vectors of important pathogens of human and animals. Therefore, their microbial carriage capacity is constantly being investigated. The aim of this study was to characterize the diversity of domestic animal pathogens in ticks collected from vegetation and the ground, from different parts of Israel. Non-engorged questing adult ticks were collected from 13 localities. A total of 1196 ticks in 131 pools-83 pools of Rhipicephalus turanicus and 48 of Rhipicephalus sanguineus (with two to ten ticks per pool)-were included in this study. In addition, 13 single free-roaming Hyalomma spp. ticks were collected. Screening by molecular techniques revealed the presence of Ehrlichia canis, Anaplasma platys, Anaplasma bovis and Babesia canis vogeli DNA in R. turanicus ticks. E. canis, A. bovis, B. canis vogeli and Candidatus Midichloria mitochondrii DNA sequences were detected in R. sanguineus ticks. Candidatus Midichloria mitochondrii DNA was also detected in Hyalomma spp. ticks. Neither Hepatozoon spp. nor Bartonella spp. DNA was detected in any of the ticks examined. This study describes the first detection of E. canis in the tick R. turanicus, which may serve as a vector of this canine pathogen; E. canis was the most common pathogen detected in the collected questing ticks. It also describes the first detection of A. bovis and Candidatus Midichloria mitochondrii in Israel. To the best of the author's knowledge, this is the first report describing the detection of DNA of the latter two pathogens in R. sanguineus, and of A. bovis in R. turanicus.

  3. Molecular cloning and characterization of a surface-localized adhesion protein in Mycoplasma bovis Hubei-1 strain.

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    Xiaohui Zou

    Full Text Available Mycoplasma bovis (M. bovis is an important pathogen that causes various bovine diseases, such as mastitis in cows and pneumonia in calves. The surface proteins are generally thought to play a central role in the pathogenesis of this organism. We screened the entire genome of M. bovis Hubei-1 and discovered a gene named vpmaX that encodes the 25 kDa variable surface lipoprotein A (VpmaX. Sequence analysis revealed that VpmaX contains several repetitive units and a typical bacterial lipoprotein signal sequence. The vpmaX gene was cloned and expressed in E. coli to obtain recombinant VpmaX (rVpmaX. Western blot analysis using a rabbit antibody against rVpmaX demonstrated that VpmaX is a membrane protein. Immunostaining visualized via confocal laser scanning microscopy showed that rVpmaX was able to adhere to embryonic bovine lung cells (EBL, and this was also confirmed by a sandwich ELISA. In summary, a surface-localized adhesion protein was identified in M. bovis Hubei-1.

  4. Molecular and serological prevalence of Babesia bovis and Babesia bigemina in water buffaloes in the northeast region of Thailand.

    Science.gov (United States)

    Terkawi, Mohamad Alaa; Huyen, Nguyen Xuan; Shinuo, Cao; Inpankaew, Tawin; Maklon, Khuanwalai; Aboulaila, Mahmoud; Ueno, Akio; Goo, Youn-Kyoung; Yokoyama, Naoaki; Jittapalapong, Sathaporn; Xuan, Xuenan; Igarashi, Ikuo

    2011-06-10

    Bovine babesiosis is a tick-transmitted hemoprotozoan disease that is mainly caused by Babesia bovis and Babesia bigemina and is characterized by significant morbidity and mortality worldwide. The disease is widespread in the northeastern region of Thailand, where an increasingly large part of the livestock is composed of water buffaloes. The present study was therefore conducted to investigate the epidemiological distribution of B. bovis and B. bigemina in water buffaloes in the northeastern region of Thailand. A total of 305 buffalo blood samples were randomly collected from five provinces and simultaneously analyzed by the nested PCR (nPCR) assay, ELISA, and IFAT techniques. The overall prevalence of B. bovis and B. bigemina was 11.2% and 3.6% by nPCR, 14.7% and 5.9% by ELISA, and 16.8% and 5.6% by IFAT, respectively. The high concordance between the molecular and the serological detection tests revealed the specificity and sensitivity of the diagnostic assays used for the detection of infection as well as the endemic stability status of the parasites in the surveyed areas. Statistically significant differences in the prevalence of the two infections were observed on the basis of age and location but not gender. Our data provide valuable information regarding the epidemiology of B. bovis and B. bigemina infection in water buffaloes in the northeastern region of Thailand which will likely be very beneficial for management and control programs of this disease.

  5. Transcriptional Response of Bovine Monocyte-Derived Macrophages after the Infection with Different Argentinean Mycobacterium bovis Isolates

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    Karina Caimi

    2013-01-01

    Full Text Available Infection of bovines with Mycobacterium bovis causes important financial hardship in many countries presenting also a risk for humans. M. bovis is known to be adapted to survive and thrive within the intramacrophage environment. In spite of its relevance, at present the information about macrophage expression patterns is scarce, particularly regarding the bovine host. In this study, transcriptomic analysis was used to detect genes differentially expressed in macrophages derived from peripheral blood mononuclear cells at early stages of infection with two Argentinean strains of M. bovis, a virulent and an attenuated strains. The results showed that the number of differentially expressed genes in the cells infected with the virulent strain (5 was significantly lower than those in the cells infected with the attenuated strain (172. Several genes were more strongly expressed in infected macrophages. Among them, we detected encoding transcription factors, anthrax toxin receptor, cell division and apoptosis regulator, ankyrin proteins, cytoskeleton proteins, protein of cell differentiation, and regulators of endocytic traffic of membrane. Quantitative real-time PCR of a selected group of differentially expressed genes confirmed the microarrays results. Altogether, the present results contribute to understanding the mechanisms involved in the early interaction of M. bovis with the bovine macrophage.

  6. Babesia bovis and B. bigemina DNA detected in cattle and ticks from Zimbabwe by polymerase chain reaction

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    I. Smeenk

    2000-07-01

    Full Text Available From blood collected from 94 cattle at 12 locations in the eastern and northeastern areas of Zimbabwe, DNA was extracted and analysed by polymerase chain reaction with primers previously reported to be specific for Babesia bigemina and Babesia bovis. Overall, DNA of Babesia bigemina was detected in the blood of 33/94 (35 % cattle and DNA from B. bovis was detected in 27/58 (47 % of cattle. The prevalence of DNA of B. bigemina was significantly higher in young animals (<2 years (23/46 than in animals over 2 years of age (10/48; (chi2 = 8.77; P < 0.01 %. Although tick sampling was not thorough, Boophilus decoloratus could be collected at 7/9 sites sampled and Boophilus microplus at 4/9 sites. Of the 20 B. decoloratus allowed to oviposit before PCR analysis, 1 (5 % contained DNA that could be amplified with primers for B. bigemina while 12 (60 % were positive with primers for B. bovis. Of the B. microplus allowed to oviposit, 11/16 (69 % were positive for B. bovis DNAby PCR and 2/16 (12 % were positive for B. bigemina.

  7. Isolation and identification of arabinose mycolates of Cell Wall Skeleton (CWS) derived from Mycobacterium bovis BCG Tokyo 172 (SMP-105).

    Science.gov (United States)

    Uenishi, Yuko; Kusunose, Naoto; Yano, Ikuya; Sunagawa, Makoto

    2010-03-01

    A unique hydrolysis method using a two-layer solution, consisting of diluted hydrochloric acid and toluene was developed to isolate whole arabinose mycolates from the cell wall skeleton of Mycobacterium bovis BCG Tokyo 172 (SMP-105) in order to reveal its pivotal role in enhancing immune responses against tumors.

  8. Species identification of Streptococcus bovis group isolates causing bacteremia: a comparison of two MALDI-TOF MS systems.

    Science.gov (United States)

    Agergaard, Charlotte N; Knudsen, Elisa; Dargis, Rimtas; Nielsen, Xiaohui C; Christensen, Jens J; Justesen, Ulrik S

    2017-02-20

    This study compared two MALDI-TOF MS systems (Biotyper and VITEK MS) on clinical Streptococcus bovis group isolates (n=66). The VITEK MS gave fewer misidentifications and a higher rate of correct identifications than the Biotyper. Only the identification of S. lutetiensis by the VITEK MS was reliable. Additional optimization of the available system databases is needed.

  9. Usefulness of restriction fragment length polymorphism and spoligotyping for epidemiological studies of Mycobacterium bovis in Madagascar: description of new genotypes.

    Science.gov (United States)

    Rasolofo Razanamparany, Voahangy; Quirin, René; Rapaoliarijaona, Andriampenomaka; Rakotoaritahina, Hugues; Vololonirina, Elie Jeanne; Rasolonavalona, Tiana; Ferdinand, Séverine; Sola, Christophe; Rastogi, Nalin; Ramarokoto, Herimanana; Chanteau, Suzanne

    2006-04-16

    Tuberculosis is highly prevalent in cattle in Madagascar. An epidemiological study based on genotyping of Mycobacterium bovis and its transmission to humans was carried out. The restriction fragment length polymorphism (IS6110 and DR markers) and spoligotyping were used to assess the genetic diversity of strains from different regions of Madagascar. One of these strains was isolated from goat, the other strains were isolated from zebu cattle. Nine IS6110 profiles, 20 DR profiles and 12 spoligotypes were obtained. About 90% of all isolates gave a single IS6110 band at about 1.8 kb. Most strains had the same spoligotype. M. bovis strains commonly lack spacers 39-43, and all Malagasy strains also lacked spacers 3-5, 8-10 and 16. This pattern has not been reported elsewhere. DR was the most discriminatory of the three markers. The patterns obtained with the three markers were combined to identify 34 different genotypes, one of which was found in 35% of the strains. No region-specific M. bovis genotype was identified, but the genotyping of 18 M. bovis strains isolated from patients showed that the human and bovine strains were identical, suggesting possible human contamination from zebu cattle.

  10. [Bacteremia and endocarditis caused by Streptococcus bovis in patients with alcoholic hepatopathy without evidence of colonic pathology].

    Science.gov (United States)

    Castroagudín, J F; Lorenzo Solar, M; Martínez Rey, C; Brage Varela, A; Torre, J A; González Quintela, A

    1996-09-01

    The association of Streptococcus bovis bacteremia and endocarditis with colonic pathology, mainly neoplastic, is well known. Its relationship with liver disease without evidence of gastrointestinal disease has been rarely described. To analyze the association between S. bovis infection and liver disease, positive blood cultures for this microorganism in hospitalized patients in the Internal Medicine and Gastroenterology Departments from December 1993 until October 1995, have been reviewed. Three cases of S. bovis infection (one bacteremia, two endocarditis) were found. Alcoholic liver disease was diagnosed in all three patients, with associated hepatitis C virus in one of them. Colonic pathology was excluded by colonoscopy and/or barium enema. Other gastrointestinal disorders were excluded by means of gastroscopy, barium gastrointestinal study and abdominal ultrasonography. Antibiotic therapy was based in betalactamics, with associated aminoglycoside in two cases. One patient needed aortic and mitral valve replacement and another one needed orthotopic liver transplantation. No new gastrointestinal pathology emerged in the follow-up (5-23 months). Cases of S. bovis bacteremia and endocarditis should be screened not also for colonic pathology, but also for liver disease, particularly in alcoholics.

  11. Reactive Nitrogen Intermediates Production by Macrophages of Mycobacterium bovis-Infected Goats and Supplemented with Dyhidroxyvitamin D3 in vivo

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    Aurora Martinez-Romero

    2009-01-01

    Full Text Available Problem statement: Tuberculosis (TB remains one of the world's major health problems. To evaluate in vivo indices of cellular sensitization, antigen-induced Reactive Nitrogen Intermediates (RNI responses by blood mononuclear cells from Mycobacterium bovis BCG-infected goats supplemented with 1, 25 dyhidroxyvitamin D3 [1, 25-(OH2D3]. Approach: An experimental, longitudinal and comparative study was planned. Twelve animals of goat cattle 20-to 24-month-old sannen selected themselves. Five samplings were made, previous to the inoculation (zero d, 3, 7, 14 and 21 d after applying the treatments. The mononuclear cells by the Ficoll-hypaque method were obtained. The RNI, nitrites and nitrates (NO2- and NO3-­ were quantified by Enzyme-Linked Immunosorbent Assay (ELISA. Results: The treatment with the 1, 25(OH2D3 stimulated the NO3- synthesis indicating, that by itself it is a good modulador of the micobacterial replication and in the treatment with M. bovis-BCG vaccine increased as a result to the treatment with 1,25(OH2D3. The exhibition to M. bovis-BCG vaccine with the treatment with 1, 25(OH2D3 was able to increase answer NO3-­ in exposed animals. Conclusion: The 1, 25(OH2D3 stimulated in vivo the production of RNI in goats exposed to M. bovis BCG vaccine.

  12. Sodium-Dependent Transport of Neutral Amino Acids by Whole Cells and Membrane Vesicles of Streptococcus bovis, a Ruminal Bacterium

    NARCIS (Netherlands)

    Russell, James B.; Strobel, Herbert J.; Driessen, Arnold J.M.; Konings, Wilhelmus

    1988-01-01

    Streptococcus bovis JB1 cells were able to transport serine, threonine, or alanine, but only when they were incubated in sodium buffers. If glucose-energized cells were washed in potassium phosphate and suspended in potassium phosphate buffer, there was no detectable uptake. Cells deenergized with 2

  13. A Moraxella bovis pili vaccine produced by recombinant DNA technology for the prevention of infectious bovine keratoconjunctivitis.

    Science.gov (United States)

    Lepper, A W; Elleman, T C; Hoyne, P A; Lehrbach, P R; Atwell, J L; Schwartzkoff, C L; Egerton, J R; Tennent, J M

    1993-07-01

    Pili (fimbriae) were prepared from Moraxella bovis strain Dalton 2d (Dal2d) and from a derivative of Pseudomonas aeruginosa K/2PfS that contained a plasmid-borne Dal2d pilin gene and produced pili having serogroup-specific identity to Dal2d. Nine calves were vaccinated with two doses each of 30 micrograms authentic M. bovis Dal2d pili in oil adjuvant and 10 calves were vaccinated with a similar dose of P. aeruginosa-derived Dal2d pili in the same formulation. All 19 calves and 10 non-vaccinated controls were challenged by instillation of 1 x 10(9) virulent M. bovis Dal2d cells into both conjunctival sacs 19 days after the second vaccine dose. The serological response to vaccination and the degree of protection against experimentally induced infectious bovine keratoconjunctivitis (IBK) were assessed. None of the nine calves vaccinated with authentic M. bovis Dal2d pili developed IBK while two of those vaccinated with P. aeruginosa-derived Dal2d pili developed lesions which accounted for a mean group lesion score of 0.3. In contrast, 9 of the 10 non-vaccinated calves developed IBK lesions, the majority of which were progressive, required early treatment and accounted for a mean group lesion score of 1.5. These results demonstrate the potential of a relatively low dose of pili produced by recombinant DNA technology for development of an effective vaccine against IBK.

  14. Expressed gene sequences from larval genes over-expressed upon Babesia bovis infection of Rhipecephalus (Boophilus) microplus

    Science.gov (United States)

    Boophilus ticks are vectors of Babesia bovis, the protozoan causative agent of cattle fever, a disease which is responsible for significant production losses to cattle producers in much of Africa, Central and South America and Australia. We utilized subtractive cDNA library synthesis techniques to o...

  15. Fasciola hepatica infection reduces Mycobacterium bovis burden and mycobacterial uptake and suppresses the pro-inflammatory response.

    Science.gov (United States)

    Garza-Cuartero, L; O'Sullivan, J; Blanco, A; McNair, J; Welsh, M; Flynn, R J; Williams, D; Diggle, P; Cassidy, J; Mulcahy, G

    2016-07-01

    Bovine tuberculosis (BTB), caused by Mycobacterium bovis, has an annual incidence in cattle of 0.5% in the Republic of Ireland and 4.7% in the UK, despite long-standing eradication programmes being in place. Failure to achieve complete eradication is multifactorial, but the limitations of diagnostic tests are significant complicating factors. Previously, we have demonstrated that Fasciola hepatica infection, highly prevalent in these areas, induced reduced sensitivity of the standard diagnostic tests for BTB in animals co-infected with F. hepatica and M. bovis. This was accompanied by a reduced M. bovis-specific Th1 immune response. We hypothesized that these changes in co-infected animals would be accompanied by enhanced growth of M. bovis. However, we show here that mycobacterial burden in cattle is reduced in animals co-infected with F. hepatica. Furthermore, we demonstrate a lower mycobacterial recovery and uptake in blood monocyte-derived macrophages (MDM) from F. hepatica-infected cattle which is associated with suppression of pro-inflammatory cytokines and a switch to alternative activation of macrophages. However, the cell surface expression of TLR2 and CD14 in MDM from F. hepatica-infected cattle is increased. These findings reflecting the bystander effect of helminth-induced downregulation of pro-inflammatory responses provide insights to understand host-pathogen interactions in co-infection.

  16. Inhibition of fructan-fermenting equine fecal bacteria and Streptococcus bovis by hops (Humulus lupulus L.) ß-acid

    Science.gov (United States)

    Aims: The goals were to determine if the '-acid from hops (Humulus lupulus L.) could be used to control fructan fermentation by equine hindgut microorganisms, and to verify the antimicrobial mode of action on the Streptococcus bovis, which has been implicated in fructan fermentation, hindgut acidos...

  17. Whole-Genome Sequences of Mycobacterium bovis Strain MbURU-001, Isolated from Fresh Bovine Infected Samples

    Science.gov (United States)

    Lasserre, Moira; Berná, Luisa; Greif, Gonzalo; Díaz-Viraqué, Florencia; Naya, Hugo; Castro-Ramos, Miguel; Juambeltz, Arturo

    2015-01-01

    Bovine tuberculosis in cattle has a high incidence in Uruguay, where it is considered a disease of national importance. We present the genome sequence of Mycobacterium bovis strain MbURU-001, isolated from pectoral lymph nodes of a bovine host from a cattle farm. PMID:26543108

  18. Tuberculosis patients co-infected with Mycobacterium bovis and Mycobacterium tuberculosis in an urban area of Brazil

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    Marcio Roberto Silva

    2013-05-01

    Full Text Available In this cross-sectional study, mycobacteria specimens from 189 tuberculosis (TB patients living in an urban area in Brazil were characterised from 2008-2010 using phenotypic and molecular speciation methods (pncA gene and oxyR pseudogene analysis. Of these samples, 174 isolates simultaneously grew on Löwenstein-Jensen (LJ and Stonebrink (SB-containing media and presented phenotypic and molecular profiles of Mycobacterium tuberculosis, whereas 12 had molecular profiles of M. tuberculosis based on the DNA analysis of formalin-fixed paraffin wax-embedded tissue samples (paraffin blocks. One patient produced two sputum isolates, the first of which simultaneously grew on LJ and SB media and presented phenotypic and molecular profiles of M. tuberculosis, and the second of which only grew on SB media and presented phenotypic profiles of Mycobacterium bovis. One patient provided a bronchial lavage isolate, which simultaneously grew on LJ and SB media and presented phenotypic and molecular profiles of M. tuberculosis, but had molecular profiles of M. bovis from paraffin block DNA analysis, and one sample had molecular profiles of M. tuberculosis and M. bovis identified from two distinct paraffin blocks. Moreover, we found a low prevalence (1.6% of M. bovis among these isolates, which suggests that local health service procedures likely underestimate its real frequency and that it deserves more attention from public health officials.

  19. Polyfunctional cytokine production by central memory T cells from cattle in response to Mycobacterium bovis infection and BCG vaccination

    Science.gov (United States)

    Polyfunctional T cells simultaneously produce IFN-gamma, IL-2 and TNF-alpha and play relevant roles in several chronic infections, including TB. Mycobacterium bovis infection of cattle elicits ex vivo polyfunctional T cell responses. Vaccine-elicited IFN-gamma Tcm (CD4 plus CD45RO plus CCR7 plus) re...

  20. Trends of Mycobacterium bovis Isolation and First-Line Anti-tuberculosis Drug Susceptibility Profile: A Fifteen-Year Laboratory-Based Surveillance.

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    Miriam Bobadilla-del Valle

    2015-09-01

    Full Text Available Mycobacterium tuberculosis causes the majority of tuberculosis (TB cases in humans; however, in developing countries, human TB caused by M. bovis may be frequent but undetected. Human TB caused by M. bovis is considered a zoonosis; transmission is mainly through consumption of unpasteurized dairy products, and it is less frequently attributed to animal-to-human or human-to-human contact. We describe the trends of M. bovis isolation from human samples and first-line drug susceptibility during a 15-year period in a referral laboratory located in a tertiary care hospital in Mexico City.Data on mycobacterial isolates from human clinical samples were retrieved from the laboratory's database for the 2000-2014 period. Susceptibility to first-line drugs: rifampin, isoniazid, streptomycin (STR and ethambutol was determined. We identified 1,165 isolates, 73.7% were M. tuberculosis and 26.2%, M. bovis. Among pulmonary samples, 16.6% were M. bovis. The proportion of M. bovis isolates significantly increased from 7.8% in 2000 to 28.4% in 2014 (X(2trend, p<0.001. Primary STR resistance was higher among M. bovis compared with M. tuberculosis isolates (10.9% vs.3.4%, p<0.001. Secondary multidrug resistance (MDR rates were 38.5% and 34.4% for M. bovis and M. tuberculosis, respectively (p = 0.637. A rising trend of primary STR monoresistance was observed for both species (3.4% in 2000-2004 vs. 7.6% in 2010-2014; p = 0.02.There is a high prevalence and a rising trend of M. bovis isolates in our region. The proportion of pulmonary M. bovis isolates is higher than in previous reports. Additionally, we report high rates of primary anti-tuberculosis resistance and secondary MDR in both M. tuberculosis and M. bovis. This is one of the largest reports on drug susceptibility of M. bovis from human samples and shows a significant proportion of first-line anti-tuberculosis drug resistance.

  1. Inactivation of Mycobacterium bovis ssp. caprae in high-temperature, short-term pasteurized pilot-plant milk.

    Science.gov (United States)

    Hammer, P; Richter, E; Rüsch-Gerdes, S; Walte, H-G C; Matzen, S; Kiesner, C

    2015-03-01

    Experiments to determine the efficacy of high temperature, short time (HTST) pasteurization of milk in terms of inactivation of pathogenic microorganisms were mainly performed between 1930 and 1960. Among the target organisms were Mycobacterium bovis and Mycobacterium tuberculosis. As a result, the Codex Alimentarius prescribes that HTST treatment of milk should lead to a significant reduction of pathogenic microorganisms during milk pasteurization. Due to the development of improved methods for the detection of survivors and of more advanced heating technology, verification of this requirement seemed to be necessary. To address recent outbreaks of tuberculosis in cattle caused by M. bovis ssp. caprae (M. caprae) in the southern regions of Germany, this organism was tested and compared with M. bovis ssp. bovis (M. bovis). Experiments were performed in a pilot plant for HTST pasteurization of milk with 3 strains of M. caprae and 1 strain of M. bovis. In preliminary trials at a fixed holding time of 25 s, the temperature at which significant inactivation occurred was 62.5°C for all strains. To determine D-values (decimal reduction times) for the inactivation kinetics, the strains were tested at 65, 62.5, and 60°C at holding times of 16.5, 25, and 35 s. At 65°C, the D-values of all strains ranged from 6.8 to 7.8 s, and at 62.5°C, D-values ranged from 14.5 to 18.1 s. Low inactivation was observed at 60°C. When the low slope of the inactivation curve allowed calculation of a D-value, these ranged from 40.8 to 129.9 s. In terms of log10 reductions, the highest values for all strains were 4.1 to 4.9 log at 65°C, with a holding time of 35 s. The tested strains of M. caprae and M. bovis showed similar low resistance to heat. Standard HTST treatment should result in a high reduction of these organisms and thus the requirements of the Codex Alimentarius for inactivation of pathogens by this process are far exceeded.

  2. Combined Bacillus licheniformis and Bacillus subtilis infection in a patient with oesophageal perforation.

    Science.gov (United States)

    Jeon, You La; Yang, John Jeongseok; Kim, Min Jin; Lim, Gayoung; Cho, Sun Young; Park, Tae Sung; Suh, Jin-Tae; Park, Yong Ho; Lee, Mi Suk; Kim, Soo Cheol; Lee, Hee Joo

    2012-12-01

    Species of the genus Bacillus are a common laboratory contaminant, therefore, isolation of these organisms from blood cultures does not always indicate infection. In fact, except for Bacillus anthracis and Bacillus cereus, most species of the genus Bacillus are not considered human pathogens, especially in immunocompetent individuals. Here, we report an unusual presentation of bacteraemia and mediastinitis due to co-infection with Bacillus subtilis and Bacillus licheniformis, which were identified by 16S RNA gene sequencing, in a patient with an oesophageal perforation.

  3. African 1, an epidemiologically important clonal complex of Mycobacterium bovis dominant in Mali, Nigeria, Cameroon, and Chad.

    Science.gov (United States)

    Müller, Borna; Hilty, Markus; Berg, Stefan; Garcia-Pelayo, M Carmen; Dale, James; Boschiroli, M Laura; Cadmus, Simeon; Ngandolo, Bongo Naré Richard; Godreuil, Sylvain; Diguimbaye-Djaibé, Colette; Kazwala, Rudovick; Bonfoh, Bassirou; Njanpop-Lafourcade, Betty M; Sahraoui, Naima; Guetarni, Djamel; Aseffa, Abraham; Mekonnen, Meseret H; Razanamparany, Voahangy Rasolofo; Ramarokoto, Herimanana; Djønne, Berit; Oloya, James; Machado, Adelina; Mucavele, Custodia; Skjerve, Eystein; Portaels, Francoise; Rigouts, Leen; Michel, Anita; Müller, Annélle; Källenius, Gunilla; van Helden, Paul D; Hewinson, R Glyn; Zinsstag, Jakob; Gordon, Stephen V; Smith, Noel H

    2009-03-01

    We have identified a clonal complex of Mycobacterium bovis present at high frequency in cattle in population samples from several sub-Saharan west-central African countries. This closely related group of bacteria is defined by a specific chromosomal deletion (RDAf1) and can be identified by the absence of spacer 30 in the standard spoligotype typing scheme. We have named this group of strains the African 1 (Af1) clonal complex and have defined the spoligotype signature of this clonal complex as being the same as the M. bovis BCG vaccine strain but with the deletion of spacer 30. Strains of the Af1 clonal complex were found at high frequency in population samples of M. bovis from cattle in Mali, Cameroon, Nigeria, and Chad, and using a combination of variable-number tandem repeat typing and spoligotyping, we show that the population of M. bovis in each of these countries is distinct, suggesting that the recent mixing of strains between countries is not common in this area of Africa. Strains with the Af1-specific deletion (RDAf1) were not identified in M. bovis isolates from Algeria, Burundi, Ethiopia, Madagascar, Mozambique, South Africa, Tanzania, and Uganda. Furthermore, the spoligotype signature of the Af1 clonal complex has not been identified in population samples of bovine tuberculosis from Europe, Iran, and South America. These observations suggest that the Af1 clonal complex is geographically localized, albeit to several African countries, and we suggest that the dominance of the clonal complex in this region is the result of an original introduction into cows naïve to bovine tuberculosis.

  4. African 1, an Epidemiologically Important Clonal Complex of Mycobacterium bovis Dominant in Mali, Nigeria, Cameroon, and Chad▿ †

    Science.gov (United States)

    Müller, Borna; Hilty, Markus; Berg, Stefan; Garcia-Pelayo, M. Carmen; Dale, James; Boschiroli, M. Laura; Cadmus, Simeon; Ngandolo, Bongo Naré Richard; Godreuil, Sylvain; Diguimbaye-Djaibé, Colette; Kazwala, Rudovick; Bonfoh, Bassirou; Njanpop-Lafourcade, Betty M.; Sahraoui, Naima; Guetarni, Djamel; Aseffa, Abraham; Mekonnen, Meseret H.; Razanamparany, Voahangy Rasolofo; Ramarokoto, Herimanana; Djønne, Berit; Oloya, James; Machado, Adelina; Mucavele, Custodia; Skjerve, Eystein; Portaels, Francoise; Rigouts, Leen; Michel, Anita; Müller, Annélle; Källenius, Gunilla; van Helden, Paul D.; Hewinson, R. Glyn; Zinsstag, Jakob; Gordon, Stephen V.; Smith, Noel H.

    2009-01-01

    We have identified a clonal complex of Mycobacterium bovis present at high frequency in cattle in population samples from several sub-Saharan west-central African countries. This closely related group of bacteria is defined by a specific chromosomal deletion (RDAf1) and can be identified by the absence of spacer 30 in the standard spoligotype typing scheme. We have named this group of strains the African 1 (Af1) clonal complex and have defined the spoligotype signature of this clonal complex as being the same as the M. bovis BCG vaccine strain but with the deletion of spacer 30. Strains of the Af1 clonal complex were found at high frequency in population samples of M. bovis from cattle in Mali, Cameroon, Nigeria, and Chad, and using a combination of variable-number tandem repeat typing and spoligotyping, we show that the population of M. bovis in each of these countries is distinct, suggesting that the recent mixing of strains between countries is not common in this area of Africa. Strains with the Af1-specific deletion (RDAf1) were not identified in M. bovis isolates from Algeria, Burundi, Ethiopia, Madagascar, Mozambique, South Africa, Tanzania, and Uganda. Furthermore, the spoligotype signature of the Af1 clonal complex has not been identified in population samples of bovine tuberculosis from Europe, Iran, and South America. These observations suggest that the Af1 clonal complex is geographically localized, albeit to several African countries, and we suggest that the dominance of the clonal complex in this region is the result of an original introduction into cows naïve to bovine tuberculosis. PMID:19136597

  5. Epidemiological tracing of bovine tuberculosis in Switzerland, multilocus variable number of tandem repeat analysis of Mycobacterium bovis and Mycobacterium caprae

    Science.gov (United States)

    Scherrer, Simone; Friedel, Ute; Frei, Daniel; Suter, Dominique; Perler, Lukas; Wittenbrink, Max M.

    2017-01-01

    Background After 15 years of absence, in 2013 bovine tuberculosis (bTB), caused by Mycobacterium (M.) bovis and M. caprae, reemerged in the Swiss dairy cattle population. In order to identify the sources of infection as well as the spread of the agents, molecular-epidemiologic tracing by MIRU-VNTR analysis in combination with spoligotyping was performed. A total of 17 M. bovis and 7 M. caprae isolates were cultured from tuberculous bovine lymph nodes and analyzed with a set of 49 genetic markers by using automated capillary electrophoresis. Results The outbreak in the western part of Switzerland was caused by M. bovis spoligotype SB0120. With the exception of four single-locus variations observed in MIRU 20, the MIRU-VNTR profiles of the 17 M. bovis isolates were identical, indicating a single source of infection. M. bovis detected in one archival bovine specimen from the outbreak region showed an identical MIRU-VNTR profile, suggesting persistence of the agent in a dairy herd for nearly fifteen years. The outbreak in the eastern part of Switzerland was caused by M. caprae spoligotype SB0418. All Swiss M. caprae isolates showed the Lechtal-type MIRU-VNTR profile, described as endemic in wild ruminants and in dairy cattle in Austrian bordering regions. This suggests the agent was most likely introduced by Swiss dairy cattle summering on Austrian pastures. Conclusions The present study is the first MIRU-VNTR analysis of Swiss bTB mycobacterial isolates. The genotyping assay was found to be highly discriminating and suitable for the epidemiological tracing of further outbreaks. These findings will contribute to the development of an international MIRU-VNTR database aiming to improve bTB surveillance. PMID:28222182

  6. Overall decrease in the susceptibility of Mycoplasma bovis to antimicrobials over the past 30 years in France.

    Directory of Open Access Journals (Sweden)

    Anne V Gautier-Bouchardon

    Full Text Available Mycoplasma (M. bovis is frequently implicated in respiratory diseases of young cattle worldwide. Today, to combat M. bovis in Europe, only antimicrobial therapy is available, but often fails, leading to important economical losses. The antimicrobial susceptibility of M. bovis is not covered by antimicrobial resistance surveillance networks. The objectives of this study were to identify resistances that were acquired over the last 30 years in France and to determine their prevalence within contemporary strains. The minimum inhibition concentration (MIC values of 12 antimicrobials, considered active on M. bovis, were compared, using an agar dilution method, between 27 and 46 M. bovis isolates respectively obtained in 1978-1979 and in 2010-2012 from 73 distinct respiratory disease outbreaks in young cattle all over France. For eight antimicrobials, resistances were proven to be acquired over the period and expressed by all contemporary strains. The increase of the MIC value that inhibited 50% of the isolates (MIC50 was: i substantial for tylosin, tilmicosin, tulathromycin and spectinomycin, from 2 to >64, 2 to >128, 16 to 128 and 4 to >64 µg/mL, respectively, ii moderate for enrofloxacin, danofloxacin, marbofloxacin and oxytetracycline, from 0.25 to 0.5, 0.25 to 0.5, 0.5 to 1, 32 to >32 µg/mL, respectively. No differences were observed for gamithromycin, tildipirosin, florfenicol and valnemulin with MIC50 of 128, 128, 8, <0.03 µg/mL, respectively. If referring to breakpoint MIC values published for respiratory bovine pathogens, all contemporary isolates would be intermediate in vivo for fluoroquinolones and resistant to macrolides, oxytetracycline, spectinomycin and florfenicol.

  7. Overall decrease in the susceptibility of Mycoplasma bovis to antimicrobials over the past 30 years in France.

    Science.gov (United States)

    Gautier-Bouchardon, Anne V; Ferré, Séverine; Le Grand, Dominique; Paoli, Agnès; Gay, Emilie; Poumarat, François

    2014-01-01

    Mycoplasma (M.) bovis is frequently implicated in respiratory diseases of young cattle worldwide. Today, to combat M. bovis in Europe, only antimicrobial therapy is available, but often fails, leading to important economical losses. The antimicrobial susceptibility of M. bovis is not covered by antimicrobial resistance surveillance networks. The objectives of this study were to identify resistances that were acquired over the last 30 years in France and to determine their prevalence within contemporary strains. The minimum inhibition concentration (MIC) values of 12 antimicrobials, considered active on M. bovis, were compared, using an agar dilution method, between 27 and 46 M. bovis isolates respectively obtained in 1978-1979 and in 2010-2012 from 73 distinct respiratory disease outbreaks in young cattle all over France. For eight antimicrobials, resistances were proven to be acquired over the period and expressed by all contemporary strains. The increase of the MIC value that inhibited 50% of the isolates (MIC50) was: i) substantial for tylosin, tilmicosin, tulathromycin and spectinomycin, from 2 to >64, 2 to >128, 16 to 128 and 4 to >64 µg/mL, respectively, ii) moderate for enrofloxacin, danofloxacin, marbofloxacin and oxytetracycline, from 0.25 to 0.5, 0.25 to 0.5, 0.5 to 1, 32 to >32 µg/mL, respectively. No differences were observed for gamithromycin, tildipirosin, florfenicol and valnemulin with MIC50 of 128, 128, 8, <0.03 µg/mL, respectively. If referring to breakpoint MIC values published for respiratory bovine pathogens, all contemporary isolates would be intermediate in vivo for fluoroquinolones and resistant to macrolides, oxytetracycline, spectinomycin and florfenicol.

  8. Comparison on the Contents of Cholic Acid in Natural Calculus Bovis and Calculus Bovis Artifactus Determined by Thin Layer Chromatography Scanning%薄层扫描法测定天然牛黄与人工牛黄胆酸含量比较

    Institute of Scientific and Technical Information of China (English)

    王志强; 刘海津

    2011-01-01

    Objective:To determine and compare the total contents of cholic acid in natural calculus bovis and calculus bovis artifactus in 6 product units. Methods: The silica G thin layer plate was used with isooctane - butyl acetate - glacial acetic acid - formic acid (8 : 4 : 2 : 1) as developing solvent, λs =380 nm,λR =650 nm,in the reflection of jagged scanning. Results:The content of cholic acid in calculus bovis artifactus was the same as that in natural calculus bovis. The content of free bile acid in calculus bovis artifactus was more than that in natural calculus bovis. Therefore, calculus bovis artifactus could be the substitute for natural calculus bovis within limits. Conclusion: Thin layer chromatography ( TLC ) scanning is a reliable method to determine the contents of cholic acid in natural calculus bovis and calculus bovis artifactus.%摘要:目的:测定天然牛黄与6个生产单位的人工牛黄中胆酸的含量总量比较。方法:硅胶G薄层板,异辛烷—醋酸丁酯—冰醋酸—甲酸(8∶4∶2∶1)为展开剂,λS= 380ntm,λR=650ntm,用反射法锯齿形扫描。结果:人工牛黄与天然牛黄中的胆酸成分相同,且人工牛黄中的游离胆汁酸比天然牛黄略高,在一定范围内,人工牛黄可替代天然牛黄。结论:薄层扫描法测定天然牛黄与人工牛黄中胆酸含量方法可靠。

  9. Rapid rebound of the Treg compartment in DEREG mice limits the impact of Treg depletion on mycobacterial burden, but prevents autoimmunity.

    Directory of Open Access Journals (Sweden)

    Luciana Berod

    Full Text Available The development of an effective vaccine against tuberculosis (Tb represents one of the major medical challenges of this century. Mycobacterium bovis Bacille Calmette-Guerin (BCG, the only vaccine available at present, is mostly effective at preventing disseminated Tb in children, but shows variable protection against pulmonary Tb, the most common form in adults. The reasons for this poor efficacy are not completely understood, but there is evidence that T regulatory cells (Tregs might be involved. Similarly, Tregs have been associated with the immunosuppression observed in patients infected with Tb and are therefore believed to play a role in pathogen persistence. Thus, Treg depletion has been postulated as a novel strategy to potentiate M. bovis BCG vaccination on one side, while on the other, employed as a therapeutic approach during chronic Tb infection. Yet since Tregs are critically involved in controlling autoimmune inflammation, elimination of Tregs may therefore also incur the danger of an excessive inflammatory immune response. Thus, understanding the dynamics and function of Tregs during mycobacterial infection is crucial to evaluate the potential of Treg depletion as a medical option. To address this, we depleted Tregs after infection with M. bovis BCG or Mycobacterium tuberculosis (Mtb using DEREG mice, which express the diphtheria toxin (DT receptor under the control of the FoxP3 locus, thereby allowing the selective depletion of FoxP3+ Tregs. Our results show that after depletion, the Treg niche is rapidly refilled by a population of DT-insensitive Tregs (diTregs and bacterial load remains unchanged. On the contrary, impaired rebound of Tregs in DEREG × FoxP3GFP mice improves pathogen burden, but is accompanied by detrimental autoimmune inflammation. Therefore, our study provides the proof-of-principle that, although a high degree of Treg depletion may contribute to the control of mycobacterial infection, it carries the risk of

  10. Increased TNF-alpha/IFN-gamma/IL-2 and decreased TNF-alpha/IFN-gamma production by central memory T cells are associated with protective responses against bovine tuberculosis following BCG vaccination

    Directory of Open Access Journals (Sweden)

    Mayara Fernanda Maggioli

    2016-10-01

    Full Text Available Central memory T cells (Tcm and polyfunctional CD4 T cell responses contribute to vaccine-elicited protection with both human and bovine tuberculosis (TB; however, their combined role in protective immunity to TB is unclear. To address this question, we evaluated polyfunctional cytokine responses by CD4 T cell effector / memory populations from bacille Calmette Guerin (BCG vaccinated and non-vaccinated calves prior to and after aerosol challenge with virulent Mycobacterium bovis. Polyfunctional cytokine expression patterns in the response by Tcm, effector memory, and effector T cell subsets were similar between BCG-vaccinated and M. bovis-infected calves; only differing in magnitude (i.e., infected > vaccinated. BCG vaccination, however, did alter the kinetics of the ensuing response to virulent M. bovis infection. Early after challenge (three weeks post-infection, non-vaccinates had greater antigen-specific IFN-γ/TNF-α and lesser IFN-γ/TNF-α/IL-2 responses by Tcm cells than did vaccinated animals. Importantly, these differences were also associated with mycobacterial burden upon necropsy. Polyfunctional responses to ESAT-6:CFP10 (antigens not synthesized by BCG strains were detected in memory subsets, as well as in effector cells, as early as three weeks after challenge. These findings suggest that cell fate divergence may occur early after antigen priming in the response to bovine TB and that memory and effector T cells may expand concurrently during the initial phase of the immune response. In summary, robust IFN-γ/TNF-α response by Tcm cells is associated with greater mycobacterial burden while IFN-γ/TNF-α/IL-2 response by Tcm cells are indicative of a protective response to bovine TB.

  11. Pathogenic Mycobacterium bovis strains differ in their ability to modulate the proinflammatory activation phenotype of macrophages

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    Andrade Marcelle RM

    2012-08-01

    Full Text Available Abstract Background Tuberculosis, caused by Mycobacterium tuberculosis or Mycobacterium bovis, remains one of the leading infectious diseases worldwide. The ability of mycobacteria to rapidly grow in host macrophages is a factor contributing to enhanced virulence of the bacteria and disease progression. Bactericidal functions of phagocytes are strictly dependent on activation status of these cells, regulated by the infecting agent and cytokines. Pathogenic mycobacteria can survive the hostile environment of the phagosome through interference with activation of bactericidal responses. To study the mechanisms employed by highly virulent mycobacteria to promote their intracellular survival, we investigated modulating effects of two pathogenic M. bovis isolates and a reference M. tuberculosis H37Rv strain, differing in their ability to multiply in macrophages, on activation phenotypes of the cells primed with major cytokines regulating proinflammatory macrophage activity. Results Bone marrow- derived macrophages obtained from C57BL/6 mice were infected by mycobacteria after a period of cell incubation with or without treatment with IFN-γ, inducing proinflammatory type-1 macrophages (M1, or IL-10, inducing anti-inflammatory type-2 cells (M2. Phenotypic profiling of M1 and M2 was then evaluated. The M. bovis strain MP287/03 was able to grow more efficiently in the untreated macrophages, compared with the strains B2 or H37Rv. This strain induced weaker secretion of proinflammatory cytokines, coinciding with higher expression of M2 cell markers, mannose receptor (MR and arginase-1 (Arg-1. Treatment of macrophages with IFN-γ and infection by the strains B2 and H37Rv synergistically induced M1 polarization, leading to high levels of inducible nitric oxide synthase (iNOS expression, and reduced expression of the Arg-1. In contrast, the cells infected with the strain MP287/03 expressed high levels of Arg-1 which competed with iNOS for the common substrate

  12. Rapid detection of Mycobacterium bovis DNA in cattle lymph nodes with visible lesions using PCR

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    Jahans Keith

    2007-06-01

    Full Text Available Abstract Background We have evaluated a sensitive screening assay for Mycobacterium tuberculosis (MTB complex organisms and a specific assay for detecting Mycobacterium bovis DNA in lymph nodes taken from cattle with evidence of bovine tuberculosis. Underlying these series of experiments was the need for a versatile DNA extraction protocol which could handle tissue samples and with the potential for automation. The target for the screening assay was the multi-copy insertion element IS1081, present in 6 copies in the MTB complex. For confirmation of M. bovis we used primers flanking a specific deletion in the genome of M. bovis known as region of difference 4 (RD4. The sensitivity and specificity of these PCRs has been tested on genomic DNA from MTB complex reference strains, mycobacteria other than tuberculosis (MOTT, spiked samples and on clinical material. Results The minimum detection limits of the IS1081 method was Initial testing of field samples of bovine lymph nodes with visible lesions (VL, n = 109 highlighted two shortfalls of the molecular approach. Firstly, comparison of IS1081 PCR with the "gold standard" of culture showed a sensitivity of approximately 70%. The sensitivity of the RD4 PCR method was 50%. Secondly, the success rate of spoligotyping applied directly to clinical material was 51% compared with cultures. A series of further experiments indicated that the discrepancy between sensitivity of detection found with purified mycobacterial DNA and direct testing of field samples was due to limited mycobacterial DNA recovery from tissue homogenates rather than PCR inhibition. The resilient mycobacterial cell wall, the presence of tissue debris and the paucibacillary nature of some cattle VL tissue may all contribute to this observation. Any of these factors may restrict application of other more discriminant typing methods. A simple means of increasing the efficiency of mycobacterial DNA recovery was assessed using a further pool

  13. Photothermal spectroscopy of Bacillus anthracis and Bacillus cereus with microcantilevers

    Energy Technology Data Exchange (ETDEWEB)

    Wig, Andrew G [ORNL; Arakawa, Edward T [ORNL; Passian, Ali [ORNL; Ferrell, Thomas L [ORNL; Thundat, Thomas George [ORNL

    2006-03-01

    Microcalorimetric optical and infrared spectroscopy is a method of determining the spectral absorption of small quantities of materials over a wide range of incident wavelengths. In this paper, the first spectroscopic results for microcantilevers coated with Bacillus anthracis (BA) are presented. These results, for B. anthracis from 2.5 to 14.5 {micro}m, are compared with results from microcantilevers coated with Bacillus cereus (BC) and standard spectroscopic absorption data. The results demonstrate strong correlation between the deflection measurements and the reference spectroscopic absorption peaks. An advantage of this microcantilever-based method over traditional spectroscopy is that much smaller amounts of material (nanogram quantities) can be detected in comparison with the milligram amounts needed for standard methods. Another advantage is that the complete system can be relatively small without sacrificing spectral resolution.

  14. FORMALDEHYDE GAS INACTIVATION OF BACILLUS ANTHRACIS, BACILLUS SUBTILIS AND GEOBACILLUS STEAROTHERMOPHILUS SPORES ON INDOOR SURFACE MATERIALS.

    Science.gov (United States)

    Research evaluated the decontamination of Bacillus anthracis, Bacillus subtilis, and Geobacillus stearothermophilus spores on indoor surface material using formaldehyde gas. Spores were dried on seven types of indoor surfaces and exposed to 1100 ppm formaldehyde gas for 10 hr. Fo...

  15. Mycobacterium bovis identification by a molecular method from post-mortem inspected cattle obtained in abattoirs of Mato Grosso do Sul, Brazil

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    Cristina Pires de Araújo

    2005-11-01

    Full Text Available The presence of Mycobacterium bovis in bovine carcasses with lesions suggestive of tuberculosis was evaluated. Seventy-two carcass samples were selected during slaughter inspection procedures in abattoirs in the state of Mato Grosso do Sul, Brazil. Seventeen (23.6% of samples showed colonies suggestive of mycobacteria that were confirmed to be acid-fast bacilli by Ziehl-Neelsen staining. Polymerase chain reaction (PCR using primers specific for M. bovis identified M. bovis in 13 (76.5% isolates. The PCR-restriction enzyme pattern analysis using gene encoding for the 65-kDa protein and two restriction enzymes identified the remaining four isolates that were represented by two M. tuberculosis complex and two nontuberculous mycobacteria. The results are indicative of infection of slaughter cattle by M. bovis and other mycobacteria in the state of Mato Grosso do Sul.

  16. Screening of Bacillus Species with Potentials of Antibiotics Production

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    Faruk Adamu KUTA

    2009-07-01

    Full Text Available Sixteen soil samples were collected from different refuse dump sites in Minna, the capital Niger State, and analysed for the presence of Bacillus species. Physical-chemical analysis of the soil samples revealed the followings: PH value 6.89-8.47; moisture content 1.58 – 21.21% and temperature 27-28ºC. Using both pour plate and streak method of inoculation, total bacterial count in the soil samples ranged from 3.8×104 cfu/g 16.0×104 cfu/g. The identified Bacillus species included: Bacillus cereus (30.8%, Bacillus brevis (1.9% Bacillus polymyxa (3.8%, Bacillus lichenifomis (13.5%, Bacillus spherericus (7.7%, Bacillus mycoides (13.5%, Bacillus pumilus (7.7%, Bacillus subtilis (3.8%, Bacillus alvei (1.9%, Bacillus laterosporous (1.9%, Bacillus firmus (9.6% and Bacillus circulars (3.8%. Antibiotic production tests indicated that nine Bacillus species out of twelve isolated in this study could be used to produce antibiotics that had effect on the test organisms. However, Bacillus polymyxa, Bacillus sphaericus and Bacillus laterosporous had little or no effect on the tested organisms. This study suggests that some Bacillus species have potential to produce high quality antibiotics that can be use to control microbial growth in future.

  17. beta-Amylase production by some Bacillus cereus, Bacillus megaterium and Bacillus polymyxa [correction of polymaxa] strains.

    Science.gov (United States)

    Niziołek, S

    1997-01-01

    The production of extracellular beta-amylase by some Bacillus cereus, Bacillus megaterium and Bacillus polymyxa [corrected] strains was investigated, and the maximal yields of the enzyme were 3.6; 9.3 and 20.4 U/mL of the culture fluid, respectively (U, 1 mumol of maltose equivalent per min at 30 degrees C). Several cultivation media were used for beta-amylase production. Bacillus cereus and some strains of Bacillus megaterium gave good yields of beta-amylase only in medium with the addition of nutrient broth. However, beta-amylase produced during growth in protein rich medium (nutrient broth) was highly unstable, probably due to inactivation by proteolytic enzymes co-existing in the culture fluid. Bacillus polymyxa [corrected] strains can produce good yields of beta-amylase on a semi-synthetic medium consisting of inorganic salts, potato starch and inexpensive soybean extract instead of costly peptone and meat extract. The most potential beta-amylase producer was the strain Bacillus polymyxa [corrected] NCIB 8524. The tested Bacillus megaterium and Bacillus polymyxa [corrected] strains were apparently differentiated by temperature cultivation (30 and 37 degrees C) suitable for beta-amylase amylase yield.

  18. The role of neutrophils and TNF-related apoptosis-inducing ligand (TRAIL) in bacillus Calmette-Guérin (BCG) immunotherapy for urothelial carcinoma of the bladder.

    Science.gov (United States)

    Rosevear, Henry M; Lightfoot, Andrew J; O'Donnell, Michael A; Griffith, Thomas S

    2009-12-01

    Intravesical Mycobacterium bovis bacillus Calmette-Guérin (BCG) immunotherapy is a highly effective treatment for carcinoma in situ of the bladder, as well as high-risk nonmuscle invasive urothelial carcinoma of the bladder. Despite over 30 years of clinical experience with BCG, the therapy's mechanism has remained enigmatic. Observations regarding the role of neutrophils in BCG immunotherapy have led to exciting discoveries regarding the potential role of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) in creating the therapeutic benefit of BCG immunotherapy. In this paper, we will review the scope of the disease, highlight our understanding of the role for BCG in urothelial carcinoma of the bladder, explain the recent discoveries regarding the role of neutrophils and TRAIL in therapy, and theorize on potential future areas of research.

  19. Global gene expression and systems biology analysis of bovine monocyte-derived macrophages in response to in vitro challenge with Mycobacterium bovis.

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    David A Magee

    Full Text Available BACKGROUND: Mycobacterium bovis, the causative agent of bovine tuberculosis, is a major cause of mortality in global cattle populations. Macrophages are among the first cell types to encounter M. bovis following exposure and the response elicited by these cells is pivotal in determining the outcome of infection. Here, a functional genomics approach was undertaken to investigate global gene expression profiles in bovine monocyte-derived macrophages (MDM purified from seven age-matched non-related females, in response to in vitro challenge with M. bovis (multiplicity of infection 2:1. Total cellular RNA was extracted from non-challenged control and M. bovis-challenged MDM for all animals at intervals of 2 hours, 6 hours and 24 hours post-challenge and prepared for global gene expression analysis using the Affymetrix® GeneChip® Bovine Genome Array. RESULTS: Comparison of M. bovis-challenged MDM gene expression profiles with those from the non-challenged MDM controls at each time point identified 3,064 differentially expressed genes 2 hours post-challenge, with 4,451 and 5,267 differentially expressed genes detected at the 6 hour and 24 hour time points, respectively (adjusted P-value threshold ≤ 0.05. Notably, the number of downregulated genes exceeded the number of upregulated genes in the M. bovis-challenged MDM across all time points; however, the fold-change in expression for the upregulated genes was markedly higher than that for the downregulated genes. Systems analysis revealed enrichment for genes involved in: (1 the inflammatory response; (2 cell signalling pathways, including Toll-like receptors and intracellular pathogen recognition receptors; and (3 apoptosis. CONCLUSIONS: The increased number of downregulated genes is consistent with previous studies showing that M. bovis infection is associated with the repression of host gene expression. The results also support roles for MyD88-independent signalling and intracellular PRRs in

  20. Genome-wide transcriptional profiling of peripheral blood leukocytes from cattle infected with Mycobacterium bovis reveals suppression of host immune genes

    OpenAIRE

    O'Farrelly, Cliona; Hokamp, Karsten

    2011-01-01

    PUBLISHED Background: Mycobacterium bovis is the causative agent of bovine tuberculosis (BTB), a pathological infection with significant economic impact. Recent studies have highlighted the role of functional genomics to better understand the molecular mechanisms governing the host immune response to M. bovis infection. Furthermore, these studies may enable the identification of novel transcriptional markers of BTB that can augment current diagnostic tests and surveillance programmes. I...

  1. Differentiation of Moraxella nonliquefaciens, M. lacunata, and M. bovis by using multilocus enzyme electrophoresis and hybridization with pilin-specific DNA probes.

    OpenAIRE

    1992-01-01

    Genetic relationships among strains of Moraxella nonliquefaciens, M. lacunata, and M. bovis were studied by using multilocus enzyme electrophoresis and DNA-DNA hybridization. The 74 isolates analyzed for electrophoretic variation at 12 enzyme loci were assigned to 59 multilocus genotypes. The multilocus genotypes were grouped in four major clusters, one representing strains of M. nonliquefaciens, two representing strains of M. lacunata, and one comprising strains of M. bovis and the single st...

  2. The role of glutamine oxoglutarate aminotransferase and glutamate dehydrogenase in nitrogen metabolism in Mycobacterium bovis BCG.

    Directory of Open Access Journals (Sweden)

    Albertus J Viljoen

    Full Text Available Recent evidence suggests that the regulation of intracellular glutamate levels could play an important role in the ability of pathogenic slow-growing mycobacteria to grow in vivo. However, little is known about the in vitro requirement for the enzymes which catalyse glutamate production and degradation in the slow-growing mycobacteria, namely; glutamine oxoglutarate aminotransferase (GOGAT and glutamate dehydrogenase (GDH, respectively. We report that allelic replacement of the Mycobacterium bovis BCG gltBD-operon encoding for the large (gltB and small (gltD subunits of GOGAT with a hygromycin resistance cassette resulted in glutamate auxotrophy and that deletion of the GDH encoding-gene (gdh led to a marked growth deficiency in the presence of L-glutamate as a sole nitrogen source as well as reduction in growth when cultured in an excess of L-asparagine.

  3. Characterization of pilin genes from seven serologically defined prototype strains of Moraxella bovis.

    Science.gov (United States)

    Atwell, J L; Tennent, J M; Lepper, A W; Elleman, T C

    1994-08-01

    Numerous field isolates of Moraxella bovis have previously been classified by serological techniques into seven serogroups, each defined by homologous cross-reaction with antisera prepared against purified pili of a single prototype strain. The gene encoding pilin from each of the prototype strains has been characterized by nucleotide sequence determination. The coding sequences show extensive homology (70 to 80%) while the proximal downstream sequences show a dichotomy into nonhomologous sets. The pilin genes of three more strains were also characterized. The presence of an additional, partial pilin gene in each prototype strain was confirmed by Southern blot analysis, and the partial pilin genes from two strains of one serogroup were characterized by sequence determination. Features of the pilin gene sequences are considered in relation to pilin gene inversion and the serological variants of strains which may arise from gene inversion events.

  4. A New Experimental Infection Model in Ferrets Based on Aerosolised Mycobacterium bovis

    Directory of Open Access Journals (Sweden)

    Lyanne McCallan

    2011-01-01

    Full Text Available There is significant interest in developing vaccines to control bovine tuberculosis, especially in wildlife species where this disease continues to persist in reservoir species such as the European Badger (Meles meles. However, gaining access to populations of badgers (protected under UK law is problematic and not always possible. In this study, a new infection model has been developed in ferrets (Mustela furo, a species which is closely related to the badger. Groups of ferrets were infected using a Madison infection chamber and were examined postmortem for the presence of tuberculous lesions and to provide tissue samples for confirmation of Mycobacterium bovis by culture. An infectious dose was defined, that establishes infection within the lungs and associated lymph nodes with subsequent spread to the mesentery lymph nodes. This model, which emphasises respiratory tract infection, will be used to evaluate vaccines for the control of bovine tuberculosis in wildlife species.

  5. Antimicrobials of Bacillus species: mining and engineering

    OpenAIRE

    Zhao, Xin

    2016-01-01

    Bacillus sp. have been successfully used to suppress various bacterial and fungal pathogens. Due to the wide availability of whole genome sequence data and the development of genome mining tools, novel antimicrobials are being discovered and updated,;not only bacteriocins, but also NRPs and PKs. A new classification system of known and putative antimicrobial compounds of Bacillus by genome mining is presented in Chapter 2. Importantly, predicting, isolating and screening of Bacillus strains w...

  6. Microarray-based Resequencing of Multiple Bacillus anthracis Isolates

    Science.gov (United States)

    2004-12-17

    al.: Iden- tification of anthrax toxin genes in a Bacillus cereus associ- ated with an illness resembling inhalation anthrax. Proc Natl Acad Sci USA...Norwegian Bacillus cereus and Bacillus thuringiensis soil isolates. Appl Environ Microbiol 2001, 67:4863-4873. 26. Radnedge L, Agron PG, Hill KK, Jackson PJ...Ticknor LO, Keim P, Andersen GL: Genome differences that distinguish Bacillus anthracis from Bacillus cereus and Bacillus thuringiensis . Appl

  7. Zoonotic aspects of Mycobacterium bovis and Mycobacterium avium-intracellulare complex (MAC).

    Science.gov (United States)

    Biet, Franck; Boschiroli, Maria Laura; Thorel, Marie Françoise; Guilloteau, Laurence A

    2005-01-01

    Pathogens that are transmitted between the environment, wildlife, livestock and humans represent major challenges for the protection of human and domestic animal health, the economic sustainability of agriculture, and the conservation of wildlife. Among such pathogens, the genus Mycobacterium is well represented by M. bovis, the etiological agent of bovine tuberculosis, M. avium ssp. paratuberculosis (Map) the etiological agent of Johne disease, M. avium ssp. avium (Maa) and in a few common cases by other emergent environmental mycobacteria. Epidemiologic surveys performed in Europe, North America and New Zealand have demonstrated the existence and importance of environmental and wildlife reservoirs of mycobacterial infections that limit the attempts of disease control programmes. The aim of this review is to examine the zoonotic aspects of mycobacteria transmitted from the environment and wildlife. This work is focused on the species of two main groups of mycobacteria classified as important pathogens for humans and animals: first, M. bovis, the causative agent of bovine tuberculosis, which belongs to the M. tuberculosis complex and has a broad host range including wildlife, captive wildlife, domestic livestock, non-human primates and humans; the second group examined, is the M. avium-intracellulare complex (MAC) which includes M. avium ssp. avium causing major health problems in AIDS patients and M. avium ssp. paratuberculosis the etiological agent of Johne disease in cattle and identified in patients with Crohn disease. MAC agents, in addition to a broad host range, are environmental mycobacteria found in numerous biotopes including the soil, water, aerosols, protozoa, deep litter and fresh tropical vegetation. This review examines the possible reservoirs of these pathogens in the environment and in wildlife, their role as sources of infection in humans and animals and their health impact on humans. The possibilities of control and management programmes for

  8. Comparative Proteomics Identifies Host Immune System Proteins Affected by Infection with Mycobacterium bovis.

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    Vladimir López

    2016-03-01

    Full Text Available Mycobacteria of the Mycobacterium tuberculosis complex (MTBC greatly impact human and animal health worldwide. The mycobacterial life cycle is complex, and the mechanisms resulting in pathogen infection and survival in host cells are not fully understood. Eurasian wild boar (Sus scrofa are natural reservoir hosts for MTBC and a model for mycobacterial infection and tuberculosis (TB. In the wild boar TB model, mycobacterial infection affects the expression of innate and adaptive immune response genes in mandibular lymph nodes and oropharyngeal tonsils, and biomarkers have been proposed as correlates with resistance to natural infection. However, the mechanisms used by mycobacteria to manipulate host immune response are not fully characterized. Our hypothesis is that the immune system proteins under-represented in infected animals, when compared to uninfected controls, are used by mycobacteria to guarantee pathogen infection and transmission. To address this hypothesis, a comparative proteomics approach was used to compare host response between uninfected (TB- and M. bovis-infected young (TB+ and adult animals with different infection status [TB lesions localized in the head (TB+ or affecting multiple organs (TB++]. The results identified host immune system proteins that play an important role in host response to mycobacteria. Calcium binding protein A9, Heme peroxidase, Lactotransferrin, Cathelicidin and Peptidoglycan-recognition protein were under-represented in TB+ animals when compared to uninfected TB- controls, but protein levels were higher as infection progressed in TB++ animals when compared to TB- and/or TB+ adult wild boar. MHCI was the only protein over-represented in TB+ adult wild boar when compared to uninfected TB- controls. The results reported here suggest that M. bovis manipulates host immune response by reducing the production of immune system proteins. However, as infection progresses, wild boar immune response recovers to

  9. Comparative Proteomics Identifies Host Immune System Proteins Affected by Infection with Mycobacterium bovis.

    Science.gov (United States)

    López, Vladimir; Villar, Margarita; Queirós, João; Vicente, Joaquín; Mateos-Hernández, Lourdes; Díez-Delgado, Iratxe; Contreras, Marinela; Alves, Paulo C; Alberdi, Pilar; Gortázar, Christian; de la Fuente, José

    2016-03-01

    Mycobacteria of the Mycobacterium tuberculosis complex (MTBC) greatly impact human and animal health worldwide. The mycobacterial life cycle is complex, and the mechanisms resulting in pathogen infection and survival in host cells are not fully understood. Eurasian wild boar (Sus scrofa) are natural reservoir hosts for MTBC and a model for mycobacterial infection and tuberculosis (TB). In the wild boar TB model, mycobacterial infection affects the expression of innate and adaptive immune response genes in mandibular lymph nodes and oropharyngeal tonsils, and biomarkers have been proposed as correlates with resistance to natural infection. However, the mechanisms used by mycobacteria to manipulate host immune response are not fully characterized. Our hypothesis is that the immune system proteins under-represented in infected animals, when compared to uninfected controls, are used by mycobacteria to guarantee pathogen infection and transmission. To address this hypothesis, a comparative proteomics approach was used to compare host response between uninfected (TB-) and M. bovis-infected young (TB+) and adult animals with different infection status [TB lesions localized in the head (TB+) or affecting multiple organs (TB++)]. The results identified host immune system proteins that play an important role in host response to mycobacteria. Calcium binding protein A9, Heme peroxidase, Lactotransferrin, Cathelicidin and Peptidoglycan-recognition protein were under-represented in TB+ animals when compared to uninfected TB- controls, but protein levels were higher as infection progressed in TB++ animals when compared to TB- and/or TB+ adult wild boar. MHCI was the only protein over-represented in TB+ adult wild boar when compared to uninfected TB- controls. The results reported here suggest that M. bovis manipulates host immune response by reducing the production of immune system proteins. However, as infection progresses, wild boar immune response recovers to limit pathogen

  10. Vaccination of cattle with Mycobacterium bovis BCG by a combination of systemic and oral routes.

    Science.gov (United States)

    Buddle, Bryce M; Denis, Michel; Aldwell, Frank E; Martin Vordermeier, H; Glyn Hewinson, R; Neil Wedlock, D

    2008-11-01

    Mycobacterium bovis bacille Calmette-Guérin (BCG) vaccine delivered to calves by the subcutaneous (s.c.) or by the oral route in a formulated lipid matrix has been previously shown to induce similar levels of protection against bovine tuberculosis. The current study was aimed at determining whether a combination of delivering BCG by s.c. and oral routes would enhance levels of protection, compared to only one route of vaccination. Forty calves were randomly divided into four groups (10/group). Calves were vaccinated with 10(6)colony forming units (CFU) of BCG Pasteur by the s.c. route or orally with 10(9)CFU BCG incorporated into a lipid formulation. One group received a combination of BCG administered by both the s.c. and oral routes and a non-vaccinated group served as a control. The two groups of calves that received s.c. BCG produced strong IFN-gamma responses in whole blood cultures stimulated with bovine purified protein derivative (PPD) 3 weeks after vaccination. Cattle vaccinated just with oral BCG in a lipid matrix produced a strong IFN-gamma response 8 weeks after vaccination, and peaking at 11 weeks after vaccination. All calves were challenged by the intratracheal route with M. bovis 15 weeks after vaccination and were euthanized and necropsied to assess protection at 17 weeks following challenge. BCG given s.c. or orally induced significant and comparable levels of protection against the virulent challenge. Vaccination of cattle by a combination of s.c./oral routes did not enhance protection beyond that achieved by s.c. or oral vaccination alone. We conclude that vaccination of cattle with BCG by a combination of routes has no beneficial additive effects, compared to a single s.c. administration of BCG or BCG given orally in a lipid formulation.

  11. Infectious bovine keratoconjunctivitis: subconjunctival administration of a Moraxella bovis pilus preparation enhances immunogenicity.

    Science.gov (United States)

    Pugh, G W; Kopecky, K E; McDonald, T J

    1985-04-01

    To compare the immune response elicited by 3 routes of vaccination, 36 calves were randomly allotted to 4 groups of 9 calves each. Group I was vaccinated subconjunctivally only. Group II was vaccinated concomitantly, both subconjunctivally and in the dewlap. Group III was vaccinated in the dewlap only. Group IV was not vaccinated and served as a virulence control for the Moraxella bovis culture. Calves in groups I, II, and III were given 2 inoculations with 14 days between inoculations. Twenty-one days after the last inoculation, the ventral conjunctival sac of all calves was instilled with cells of a virulent M bovis strain. After challenge exposure, all vaccinated calves (groups I, II, and III) had evidence of enhanced resistance, compared with the nonvaccinated calves. The highest to lowest gradients of immune responsiveness were: Group I greater than or equal to group II greater than group III greater than group IV. When immune criteria, such as the percentage of diseased eyes, the mean duration of infection, the severity of corneal lesions, and the serologic response were compared, groups I and II were significantly (P less than 0.05) more resistant to challenge exposure than were groups III and IV. Group III was significantly (P less than 0.05) different from group IV in severity of lesions and in serologic response. Also, the mean duration of infection was shorter, and the percentage of diseased eyes was less in group III than in group IV (P greater than 0.10). Group I was more resistant than was group II (P less than 0.10).(ABSTRACT TRUNCATED AT 250 WORDS)

  12. Natural killer cell cytokine response to M. bovis BCG Is associated with inhibited proliferation, increased apoptosis and ultimate depletion of NKp44(+CD56(bright cells.

    Directory of Open Access Journals (Sweden)

    Damien Portevin

    Full Text Available Mycobacterium bovis BCG, a live attenuated strain of M. bovis initially developed as a vaccine against tuberculosis, is also used as an adjuvant for immunotherapy of cancers and for treatment of parasitic infections. The underlying mechanisms are thought to rely on its immunomodulatory properties including the recruitment of natural killer (NK cells. In that context, we aimed to study the impact of M. bovis BCG on NK cell functions. We looked at cytotoxicity, cytokine production, proliferation and cell survival of purified human NK cells following exposure to single live particles of mycobacteria. We found that M. bovis BCG mediates apoptosis of NK cells only in the context of IL-2 stimulation during which CD56(bright NK cells are releasing IFN-γ in response to mycobacteria. We found that the presence of mycobacteria prevented the IL-2 induced proliferation and surface expression of NKp44 receptor by the CD56(bright population. In summary, we observed that M. bovis BCG is modulating the functions of CD56(bright NK cells to drive this subset to produce IFN-γ before subsequent programmed cell death. Therefore, IFN-γ production by CD56(bright cells constitutes the main effector mechanism of NK cells that would contribute to the benefits observed for M. bovis BCG as an immunotherapeutic agent.

  13. Resposta imune específica de bovinos experimentalmente sensibilizados com inóculos inativados de Mycobacterium bovis e Mycobacterium avium Specific immune response of cattle to experimental sensibilization by inactivated Mycobacterium bovis and Mycobacterium avium

    Directory of Open Access Journals (Sweden)

    Robson F.C. Almeida

    2006-12-01

    Full Text Available O diagnóstico presuntivo da tuberculose bovina é baseado na análise da resposta imune celular a antígenos micobacterianos. Procedeu-se à simulação experimental de sensibilização por Mycobacterium bovis e Mycobacterium avium inativados em bovinos a fim de acompanhar a resposta imune a partir do teste cervical comparativo e da evolução da produção específica de interferon-gama, além de identificar a interferência de reações inespecíficas por M. avium nos resultados dos testes. Verificou-se que os animais desencadearam resposta de hipersensibilidade tardia contra os bacilos inativados, e que ambos os testes diagnósticos da tuberculose bovina foram eficientes na identificação dos animais sensibilizados com M. bovis e na discriminação das reações geradas pela inoculação dos bovinos com M. avium.The presumptive diagnosis of bovine tuberculosis is based on analysis of the immune response to micobacterial antigens. This experimental simulation of sensibilization by Mycobacterium bovis and Mycobacterium avium in cattle aimed to verify the immune response by both the cervical comparative test and the evolution of the specific production of gamma-interferon, and also to identify interference of unspecified reactions by M. avium on the test results. The results support that the experimental animals started a response of delayed hypersensitivity to the inactivated bacilli, and that both diagnostic tests for bovine tuberculosis were efficient for the identification of animals sensitized with M. bovis and for discrimination of reactions generated by inoculation of cattle with M. avium.

  14. Assessment of different formulations of oral Mycobacterium bovis Bacille Calmette-Guérin (BCG) vaccine in rodent models for immunogenicity and protection against aerosol challenge with M. bovis.

    Science.gov (United States)

    Clark, Simon; Cross, Martin L; Smith, Alan; Court, Pinar; Vipond, Julia; Nadian, Allan; Hewinson, R Glyn; Batchelor, Hannah K; Perrie, Yvonne; Williams, Ann; Aldwell, Frank E; Chambers, Mark A

    2008-10-29

    Bovine tuberculosis (bTB) caused by infection with Mycobacterium bovis is causing considerable economic loss to farmers and Government in the United Kingdom as its incidence is increasing. Efforts to control bTB in the UK are hampered by the infection in Eurasian badgers (Meles meles) that represent a wildlife reservoir and source of recurrent M. bovis exposure to cattle. Vaccination of badgers with the human TB vaccine, M. bovis Bacille Calmette-Guérin (BCG), in oral bait represents a possible disease control tool and holds the best prospect for reaching badger populations over a wide geographical area. Using mouse and guinea pig models, we evaluated the immunogenicity and protective efficacy, respectively, of candidate badger oral vaccines based on formulation of BCG in lipid matrix, alginate beads, or a novel microcapsular hybrid of both lipid and alginate. Two different oral doses of BCG were evaluated in each formulation for their protective efficacy in guinea pigs, while a single dose was evaluated in mice. In mice, significant immune responses (based on lymphocyte proliferation and expression of IFN-gamma) were only seen with the lipid matrix and the lipid in alginate microcapsular formulation, corresponding to the isolation of viable BCG from alimentary tract lymph nodes. In guinea pigs, only BCG formulated in lipid matrix conferred protection to the spleen and lungs following aerosol route challenge with M. bovis. Protection was seen with delivery doses in the range 10(6)-10(7) CFU, although this was more consistent in the spleen at the higher dose. No protection in terms of organ CFU was seen with BCG administered in alginate beads or in lipid in alginate microcapsules, although 10(7) in the latter formulation conferred protection in terms of increasing body weight after challenge and a smaller lung to body weight ratio at necropsy. These results highlight the potential for lipid, rather than alginate, -based vaccine formulations as suitable delivery

  15. A selective chromogenic agar that distinguishes Bacillus anthracis from Bacillus cereus and Bacillus thuringiensis.

    Science.gov (United States)

    Juergensmeyer, Margaret A; Gingras, Bruce A; Restaino, Lawrence; Frampton, Elon W

    2006-08-01

    A selective and differential plating medium, R & F anthracis chromogenic agar (ACA), has been developed for isolating and identifying presumptive colonies of Bacillus anthracis. ACA contains the chromogenic substrate 5-bromo-4-chloro-3-indoxyl-choline phosphate that upon hydrolysis yields teal (blue green) colonies indicating the presence of phosphatidylcholine-specific phospholipase C (PC-PLC) activity. Among seven Bacillus species tested on ACA, only members of the Bacillus cereus group (B. anthracis, B. cereus, and B. thuringiensis) produced teal colonies (PC-PLC positive) having cream rings. Examination of colony morphology in 18 pure culture strains of B. anthracis (15 ATCC strains plus AMES-1-RIID, ANR-1, and AMED-RIID), with one exception, required 48 h at 35 to 37 degrees C for significant color production, whereas only 24 h was required for B. cereus and B. thuringiensis. This differential rate of PC-PLC synthesis in B. anthracis (due to the truncated plcR gene and PlcR regulator in B. anthracis) allowed for the rapid differentiation on ACA of presumptive colonies of B. anthracis from B. cereus and B. thuringiensis in both pure and mixed cultures. Effective recovery of B. anthracis from a variety of matrices having both high (soil and sewage) and low microbial backgrounds (cloth, paper, and blood) spiked with B. anthracis ANR-1 spores suggests the probable utility of ACA plating for B. anthracis recovery in a diversity of applications.

  16. Role of a bacillus Calmette-Guérin fibronectin attachment protein in BCG-induced antitumor activity.

    Science.gov (United States)

    Zhao, W; Schorey, J S; Bong-Mastek, M; Ritchey, J; Brown, E J; Ratliff, T L

    2000-04-01

    Intravesical Mycobacterium bovis bacillus Calmette-Gu*erin (BCG) is the treatment of choice for superficial bladder cancer. Previous studies showed that attachment of BCG to fibronectin within the bladder was necessary for mediation of the antitumor response. Further studies identified a bacterial receptor, fibronectin attachment protein (FAP), as an important mediator of BCG attachment to fibronectin. In vitro studies showed that a stable BCG/fibronectin interaction was dependent on FAP binding to fibronectin; however, no role for FAP in the attachment of BCG in vivo has been characterized. We now report the cloning of the M. bovis BCG FAP (FAP-B) and demonstrate an important role for FAP in the in vivo attachment of BCG to the bladder wall and in the induction of BCG-mediated antitumor activity. The predicted amino acid sequence for FAP-B shows 61% and 71% homology, respectively, with Mycobacterium avium FAP (FAP-A) and Mycobacterium leprae FAP (FAP-L). Rabbit polyclonal antibodies against Mycobacterium vaccae FAP (FAP-V) reacted with all 3 recombinant FAP proteins on Western blots. Functional studies show FAP-B to bind fibronectin via the highly conserved attachment regions previously identified for FAP-A and FAP-L and also to competitively inhibit attachment of BCG to matrix fibronectin. In vivo studies show FAP to be a necessary protein for the stable attachment of BCG to the bladder wall. Moreover, stable binding of BCG via FAP was shown to be necessary for the expression of BCG-induced antitumor activity. Our results demonstrate a biological role for FAP in the mediation of BCG-induced antitumor activity.

  17. Risk factors for Mycoplasma bovis-associated disease in farmed bison (Bison bison) herds in western Canada: A case-control study.

    Science.gov (United States)

    Bras, Ana L; Barkema, Herman W; Woodbury, Murray; Ribble, Carl; Perez-Casal, Jose; Windeyer, M Claire

    2016-07-01

    North American bison producers have been attempting to control and prevent Mycoplasma bovis-associated disease without the benefit of bison-specific knowledge. The objective of this study was to determine the clinical presentation of disease associated with M. bovis infection in western Canadian farmed bison, and to identify herd-level risk factors for M. bovis-associated disease. Bison producers (n=49) from western Canada (Manitoba, Saskatchewan, Alberta, and British Columbia) were selected for a 1:2 case-control study. Data were collected by an in-person interview using a questionnaire regarding clinical presentations of outbreaks and herd-level management factors. Risk factors associated with M. bovis outbreaks were identified using multivariable logistic regression analysis. All 17 case herds had a laboratory-confirmed diagnosis of M. bovis infection within the last 5 years. In 11 (65%) of the 17 case herds, disease associated with M. bovis infection recurred in subsequent years. Overall, 88% of case herds had recently introduced bison that later developed clinical signs associated with M. bovis infection. Within a bison operation, a median of 8% (Inter Quartile Range [IQR]: 3-11%) developed clinical signs: lameness, reluctance to move, swollen joints, difficulty breathing, coughing, sluggishness, and loss of body condition. Also, calving percentage the year after the first M. bovis outbreak was lower than calving percentage the year before the outbreak. Herd-level mortality risk during the first M. bovis outbreak in case herds ranged from 0.5 to 50% (median 5%, IQR: 3-10%) and the median case fatality risk was 100%. Case herds were more likely than control herds to have a feedlot unit (OR=7), to receive regular visits from rental trailers or trailers from other farms (OR=15), to annually vaccinate bison (OR=7), and to lose at least one bison due to fatal respiratory disease in the previous year (OR=9). These findings will aid development of evidence

  18. Serologic tests for detecting antibodies against Mycobacterium bovis and Mycobacterium avium subspecies paratuberculosis in Eurasian wild boar (Sus scrofa scrofa).

    Science.gov (United States)

    Boadella, Mariana; Lyashchenko, Konstantin; Greenwald, Reena; Esfandiari, Javan; Jaroso, Raquel; Carta, Tania; Garrido, Joseba M; Vicente, Joaquín; de la Fuente, José; Gortázar, Christian

    2011-01-01

    New tools to detect exposure of free-range Eurasian wild boar (Sus scrofa scrofa) to pathogenic mycobacteria would be valuable for improved disease surveillance and wildlife management. Two hundred sera from wild boar of known Mycobacterium bovis infection status were used to evaluate test suitability for the detection of antibodies against M. bovis and Mycobacterium avium subsp. paratuberculosis (or cross-reacting members of the M. avium complex). Two traditional enzyme-linked immunosorbent assays were evaluated using M. bovis purified protein derivative (bPPD) and paratuberculosis protoplasmatic antigen 3 (PPA3) as antigens, respectively, and a new point-of-care test format for bovine tuberculosis (bTB) that uses the innovative dual-path platform (DPP TB) test. The effect of individual factors (sex, age, lesions) on the diagnostic performance of the serologic tests was also determined. Although the DPP had a sensitivity of 89.6% and a specificity of 90.4%, for bPPD, the sensitivity was 79.2% and the specificity 100%. Both tests had a kappa agreement of 0.80. Sixty-five of 68 (95.6%) wild boar sera with antibodies against the PPA3 antigen corresponded to known M. bovis-infected wild boar. Significant differences were not observed in the bPPD and DPP readings among lesion categories or between age classes. A slight sex-related difference in sensitivity toward males in the DPP was found, but it was not detected in the bPPD enzyme-linked immunosorbent assay. The results support the use of antibody-based diagnostic tests for both large-scale and individual bTB testing of Eurasian wild boar and suggest that wild boar cannot be used as sentinels for infections caused by M. avium complex members.

  19. Prevalence of latent and active tuberculosis among dairy farm workers exposed to cattle infected by Mycobacterium bovis.

    Directory of Open Access Journals (Sweden)

    Pedro Torres-Gonzalez

    Full Text Available BACKGROUND: Human tuberculosis caused by M. bovis is a zoonosis presently considered sporadic in developed countries, but remains a poorly studied problem in low and middle resource countries. The disease in humans is mainly attributed to unpasteurized dairy products consumption. However, transmission due to exposure of humans to infected animals has been also recognized. The prevalence of tuberculosis infection and associated risk factors have been insufficiently characterized among dairy farm workers (DFW exposed in settings with poor control of bovine tuberculosis. METHODOLOGY/PRINCIPAL FINDINGS: Tuberculin skin test (TST and Interferon-gamma release assay (IGRA were administered to 311 dairy farm and abattoir workers and their household contacts linked to a dairy production and livestock facility in Mexico. Sputa of individuals with respiratory symptoms and samples from routine cattle necropsies were cultured for M. bovis and resulting spoligotypes were compared. The overall prevalence of latent tuberculosis infection (LTBI was 76.2% (95% CI, 71.4-80.9% by TST and 58.5% (95% CI, 53.0-64.0% by IGRA. Occupational exposure was associated to TST (OR 2.72; 95% CI, 1.31-5.64 and IGRA (OR 2.38; 95% CI, 1.31-4.30 adjusting for relevant variables. Two subjects were diagnosed with pulmonary tuberculosis, both caused by M. bovis. In one case, the spoligotype was identical to a strain isolated from bovines. CONCLUSIONS: We documented a high prevalence of latent and pulmonary TB among workers exposed to cattle infected with M. bovis, and increased risk among those occupationally exposed in non-ventilated spaces. Interspecies transmission is frequent and represents an occupational hazard in this setting.

  20. Field-isolated genotypes of Mycobacterium bovis vary in virulence and influence case pathology but do not affect outbreak size.

    Directory of Open Access Journals (Sweden)

    David M Wright

    Full Text Available Strains of many infectious agents differ in fundamental epidemiological parameters including transmissibility, virulence and pathology. We investigated whether genotypes of Mycobacterium bovis (the causative agent of bovine tuberculosis, bTB differ significantly in transmissibility and virulence, combining data from a nine-year survey of the genetic structure of the M. bovis population in Northern Ireland with detailed records of the cattle population during the same period. We used the size of herd breakdowns as a proxy measure of transmissibility and the proportion of skin test positive animals (reactors that were visibly lesioned as a measure of virulence. Average breakdown size increased with herd size and varied depending on the manner of detection (routine herd testing or tracing of infectious contacts but we found no significant variation among M. bovis genotypes in breakdown size once these factors had been accounted for. However breakdowns due to some genotypes had a greater proportion of lesioned reactors than others, indicating that there may be variation in virulence among genotypes. These findings indicate that the current bTB control programme may be detecting infected herds sufficiently quickly so that differences in virulence are not manifested in terms of outbreak sizes. We also investigated whether pathology of infected cattle varied according to M. bovis genotype, analysing the distribution of lesions recorded at post mortem inspection. We concentrated on the proportion of cases lesioned in the lower respiratory tract, which can indicate the relative importance of the respiratory and alimentary routes of infection. The distribution of lesions varied among genotypes and with cattle age and there were also subtle differences among breeds. Age and breed differences may be related to differences in susceptibility and husbandry, but reasons for variation in lesion distribution among genotypes require further investigation.

  1. An oral Mycobacterium bovis BCG vaccine for wildlife produced in the absence of animal-derived reagents.

    Science.gov (United States)

    Cross, Martin L; Lambeth, Matthew R; Aldwell, Frank E

    2009-09-01

    Cultures of Mycobacterium bovis BCG, comprising predominantly single-cell bacilli, were prepared in broth without animal-derived reagents. When formulated into a vegetable-derived lipid matrix, the vaccine was stable in vitro and was immunogenic in vivo upon feeding it to mice. This formulation could be useful for oral vaccination of wildlife against tuberculosis, where concern over transmissible prions may preclude the field use of vaccines containing animal products.

  2. INFLUENCE OF SURFACE-ACTIVE AGENT CONCENTRATION ON THE SPECTROPHOTOMETRIC BIOMASS ESTIMATION DURING SUBMERGED GROWTH OF Mycobacterium bovis

    Directory of Open Access Journals (Sweden)

    Sakai M.C.

    1997-01-01

    Full Text Available The influence of the surface-active agent Tween 80 (polyoxyethylene 20 sorbitan monoleate on the dispersed growth of Mycobacterium bovis - BCG - was studied. The slopes of the bacterial dry mass / absorbance curves, were significantly reduced, by the increase of the Tween 80 concentration in the medium. Such an influence was probably due to the effect Tween 80 had on the average size of bacterial clumps during the culture carried out in a bench fermenter

  3. Detection of Mycobacterium bovis in bovine and bubaline tissues using nested-PCR for TbD1.

    Directory of Open Access Journals (Sweden)

    Cristina P Araújo

    Full Text Available In the present study, a nested-PCR system, targeting the TbD1 region, involving the performance of conventional PCR followed by real-time PCR, was developed to detect Mycobacterium bovis in bovine/bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. In terms of analytical sensitivity, the DNA of M. bovis AN5 was detected up to 1.56 ng with conventional PCR, 97.6 pg with real-time PCR, and 1.53 pg with nested-PCR in the reaction mixture. The nested-PCR exhibited 100% analytical specificity for M. bovis when tested with the DNA of reference strains of environmental mycobacteria and closely-related Actinomycetales. A clinical sensitivity value of 76.0% was detected with tissue samples from animals that exhibited positive results in the comparative intradermal tuberculin test (CITT, as well as from those with lesions compatible with tuberculosis (LCT that rendered positive cultures. A clinical specificity value of 100% was detected with tissue samples from animals with CITT- results, with no visible lesions (NVL and negative cultures. No significant differences were found between the nested-PCR and culture in terms of detecting CITT+ animals with LCT or with NVL. No significant differences were recorded in the detection of CITT- animals with NVL. However, nested-PCR detected a significantly higher number of positive animals than the culture in the group of animals exhibiting LCT with no previous records of CITT. The use of the nested-PCR assay to detect M. bovis in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis.

  4. First-time detection of Mycobacterium bovis in livestock tissues and milk in the West Bank, Palestinian Territories.

    Directory of Open Access Journals (Sweden)

    Suheir Ereqat

    Full Text Available BACKGROUND: Bovine tuberculosis, bTB, is classified by the WHO as one of the seven neglected zoonontic diseases that cause animal health problems and has high potential to infect humans. In the West Bank, bTB was not studied among animals and the prevalence of human tuberculosis caused by M. bovis is unknown. Therefore, the aim of this study was to estimate the prevalence of bTB among cattle and goats and identify the molecular characteristics of bTB in our area. METHODOLOGY/PRINCIPAL FINDINGS: A total of 208 tissue samples, representing 104 animals, and 150 raw milk samples, obtained from cows and goats were examined for the presence of mycobacteria. The tissue samples were collected during routine meat inspection from the Jericho abattoir. DNA was extracted from all samples, milk and tissue biopsies (n = 358, and screened for presence of TB DNA by amplifying a 123-bp segment of the insertion sequence IS6110. Eight out of 254 animals (3.1% were found to be TB positive based on the IS6110-PCR. Identification of M. bovis among the positive TB samples was carried out via real time PCR followed by high resolution melt curve analysis, targeting the A/G transition along the oxyR gene. Spoligotyping analysis revealed a new genotype of M. bovis that was revealed from one tissue sample. SIGNIFICANCE: Detection of M. bovis in tissue and milk of livestock suggests that apparently healthy cattle and goats are a potential source of infection of bTB and may pose a risk to public health. Hence, appropriate measures including meat inspection at abattoirs in the region are required together with promotion of a health campaign emphasizing the importance of drinking pasteurized milk. In addition, further studies are essential at the farm level to determine the exact prevalence of bTB in goats and cattle herds in the West Bank and Israel.

  5. Pacemaker-associated Bacillus cereus endocarditis.

    Science.gov (United States)

    Barraud, Olivier; Hidri, Nadia; Ly, Kim; Pichon, Nicolas; Manea, Petrus; Ploy, Marie-Cécile; Garnier, Fabien

    2012-11-01

    We report the case of a pacemaker-associated Bacillus cereus endocarditis in a nonimmunocompromised patient. Antibiotic treatment was ineffective, and the pacemaker had to be removed. B. cereus was cultured from several blood samples and from the pacemaker electrodes. This case underlines the contribution of the rpoB gene for Bacillus species determination.

  6. Integration of a transfected gene into the genome of Babesia bovis occurs by legitimate homologous recombination mechanisms.

    Science.gov (United States)

    Suarez, Carlos E; Johnson, Wendell C; Herndon, David R; Laughery, Jacob M; Davis, William C

    2015-08-01

    This study examines the patterns of gene integration of gfp-bsd upon stable transfection into the T3Bo strain of Babesia bovis using a plasmid designed to integrate homologous sequences of the parasite's two identical ef-1α A and B genes. While the transfected BboTf-149-6 cell line displayed two distinct patterns of gene integration, clonal lines derived from this strain by cell sorting contained only single gfp-bsd insertions. Whole genome sequencing of two selected clonal lines, E9 and C6, indicated two distinct patterns of gfp-bsd insertion occurring by legitimate homologous recombination mechanisms: one into the expected ef-1α orf B, and another into the ef-1α B promoter. The data suggest that expression of the ef-1α orf B is not required for development of B. bovis in cultured erythrocyte stages. Use of legitimate homologous recombination mechanisms in transfected B. bovis supports the future use of transfection methods for developing efficient gene function assignment experiments using gene knockout techniques.

  7. Development and Application of a PCR Approach for Detection of Bovis, Sheep, Pig, and Chicken Derived Materials in Feedstuff

    Institute of Scientific and Technical Information of China (English)

    LUO Jia-qin; WANG Jia-qi; BU Deng-pan; LI Dan; WANG Li; WEI Hong-yang; ZHOU Ling-yun

    2008-01-01

    A PCR method for detection of bovis, sheep, pig, and chicken derived materials in feedstuff was established, and the existing method was improved according to the research on general primer and species-specific primers. First, general primer designed according to 16S rRNA gene sequence of bovis, sheep, pig, chicken, fish, and horse mtDNA was used for primary detection of animal derived materials in feedstuff. Species-specific primers designed according to conserved sequence of mtDNA of bovis, sheep, pig, and chicken were used for amplification of a 271, 274, 149, and 266 bp fragment, respectively. Further confirmation of the detection result was then carried out. PCR method for detection of animal derived materials in unknown feedstuff was developed by using general primer, relevant PCR system, and PCR condition. Also a PCR method for detection of each species (bovines, sheep, pig, and chicken) was designed by using our species-specific primers. High sensitivity and specificity of our method were confirmed with a minimum detection level of 0.1%. Method for detection of animal derived materials in this research is not only cheap and easy for operation but also precise and reliable results can be obtained. It could be one of the effective methods for the detection of animal derived materials in feedstuff.

  8. The toxicity of rifampicin polylactic acid nanoparticles against Mycobacterium bovis BCG and human macrophage THP-1 cell line

    Science.gov (United States)

    Erokhina, M.; Rybalkina, E.; Barsegyan, G.; Onishchenko, G.; Lepekha, L.

    2015-11-01

    Tuberculosis is rapidly becoming a major health problem. The rise in tuberculosis incidence stimulates efforts to develop more effective delivery systems for the existing antituberculous drugs while decreasing the side effects. The nanotechnology may provide novel drug delivery tools allowing controlled drug release. Rifampicin is one of the main antituberculous drugs, characterized by high toxicity, and Poly (L-lactic acid) (PLLA) is a biodegradable polymer used for the preparation of encapsulated drugs. The aim of our work was to evaluate the toxicity of rifampicin-PLLA nanoparticles against Mycobacterium bovis BCG using human macrophage THP-1 cell line. Our data demonstrate that rifampicin-PLLA is effective against M. bovis BCG in the infected macrophages. The drug is inducing the dysfunction of mitochondria and apoptosis in the macrophages and is acting as a potential substrate of Pgp thereby modulating cell chemosensitivity. The severity of the toxic effects of the rifampicin-PLLA nanoparticles is increasing in a dose-dependent manner. We suggest that free rifampicin induces death of M. bovis BCG after PLLA degradation and diffusion from phago-lysosomes to cytoplasm causing mitochondria dysfunction and affecting the Pgp activity.

  9. Identification of novel immunogenic proteins from Mycoplasma bovis and establishment of an indirect ELISA based on recombinant E1 beta subunit of the pyruvate dehydrogenase complex.

    Directory of Open Access Journals (Sweden)

    Zhenhong Sun

    Full Text Available The pathogen Mycoplasma bovis (M. bovis is a major cause of respiratory disease, mastitis, and arthritis in cattle. Screening the key immunogenic proteins and updating rapid diagnostic techniques are necessary to the prevention and control of M. bovis infection. In this study, 19 highly immunogenic proteins from M. bovis strain PD were identified using 2-dimensional gel electrophoresis, immunoblotting and MALDI-TOF/TOF MS. Of these 19 proteins, pyruvate dehydrogenase E1 component beta subunit (PDHB showed excellent immune reactivity and repeatability. PDHB was found to be conserved in different M. bovis isolates, as indicated by Western blot analysis. On the basis of these results, a rPDHB-based indirect ELISA (iELISA was established for the detection of serum antibodies using prokaryotically expressed recombinant PDHB protein as the coating antigen. The specificity analysis result showed that rPDHB-based iELISA did not react with other pathogens assessed in our study except M. agalactiae (which infects sheep and goats. Moreover, 358 serum samples from several disease-affected cattle feedlots were tested using this iELISA system and a commercial kit, which gave positive rates of 50.8% and 39.9%, respectively. The estimated Kappa agreement coefficient between the two methods was 0.783. Notably, 39 positive serum samples that had been missed by the commercial kit were all found to be positive by Western blot analysis. The detection rate of rPDHB-based iELISA was significantly higher than that of the commercial kit at a serum dilution ratio of 1∶5120 to 1∶10,240 (P<0.05. Taken together, these results provide important information regarding the novel immunogenic proteins of M. bovis. The established rPDHB-based iELISA may be suitable for use as a new method of antibody detection in M. bovis.

  10. Tuberculosis due to Mycobacterium bovis in humans in the south-west region of Ireland: is there a relationship with infection prevalence in cattle?

    LENUS (Irish Health Repository)

    Cotter, T P

    2012-02-03

    OBJECTIVE: To compare the incidence of tuberculosis due to Mycobacterium bovis in humans to the prevalence of M. bovis infection in cattle in south-west Ireland and discuss possible links between them. SETTING: In the south-west region of Ireland, a mixed urban and rural community (pop. 536,000), there is a residuum of human tuberculosis caused by M. bovis. METHODS: A retrospective analysis of the incidence of culture-positive M. bovis disease in humans in south-west Ireland from 1983 to 1994 and of the results of tuberculin testing in cattle from 1978 to 1994 for the same region. RESULTS: One to five cases of human tuberculosis due to M. bovis were recorded per year while the overall prevalence of bovine infection fell gradually during the period of study from 467 tuberculin-positive animals per 100,000 cattle tested in 1983 to 158 per 100,000 in 1994. CONCLUSION: The low incidence plateau of human tuberculosis due to M. bovis together with the decline in prevalence of animal infection in the overall period studied suggest a cut-off in the animal to human chain of infection at two points; the animal source and the ingestion of (now pasteurized) milk. This would suggest that disease in humans is now due to reactivation of previous foci of infection which were acquired when milk pasteurization was not compulsory. Based on this, we would anticipate a further reduction and possible elimination of human tuberculosis due to M. bovis in this region in the next 10-15 years.

  11. Tonsils of the Soft Palate Do Not Mediate the Response of Pigs to Oral Vaccination with Heat-Inactivated Mycobacterium bovis

    Science.gov (United States)

    Beltrán-Beck, Beatriz; Romero, Beatriz; Boadella, Mariana; Casal, Carmen; Bezos, Javier; Mazariegos, María; Martín, MariPaz; Galindo, Ruth C.; Pérez de la Lastra, José M.; Villar, Margarita; Garrido, Joseba M.; Sevilla, Iker A.; Asensio, Fernando; Sicilia, Javier; Lyashchenko, Konstantin P.; Domínguez, Lucas; Juste, Ramón A.; de la Fuente, José

    2014-01-01

    Mycobacterium bovis causes animal tuberculosis (TB) in cattle, humans, and other mammalian species, including pigs. The goal of this study was to experimentally assess the responses of pigs with and without a history of tonsillectomy to oral vaccination with heat-inactivated M. bovis and challenge with a virulent M. bovis field strain, to compare pig and wild boar responses using the same vaccination model as previously used in the Eurasian wild boar (Sus scrofa), to evaluate the use of several enzyme-linked immunosorbent assays (ELISAs) and lateral flow tests for in vivo TB diagnosis in pigs, and to verify if these tests are influenced by oral vaccination with inactivated M. bovis. At necropsy, the lesion and culture scores were 20% to 43% higher in the controls than those in the vaccinated pigs. Massive M. bovis growth from thoracic tissue samples was observed in 4 out of 9 controls but in none of the 10 vaccinated pigs. No effect of the presence or absence of tonsils was observed on these scores, suggesting that tonsils are not involved in the protective response to this vaccine in pigs. The serum antibody levels increased significantly only after challenge. At necropsy, the estimated sensitivities of the ELISAs and dual path platform (DPP) assays ranged from 89% to 94%. In the oral mucosa, no differences in gene expression were observed in the control group between the pigs with and without tonsils. In the vaccinated group, the mRNA levels for chemokine (C-C motif) receptor 7 (CCR7), interferon beta (IFN-β), and methylmalonyl coenzyme A mutase (MUT) were higher in pigs with tonsils. Complement component 3 mRNA levels in peripheral blood mononuclear cells (PBMC) increased with vaccination and decreased after M. bovis challenge. This information is relevant for pig production in regions that are endemic for M. bovis and for TB vaccine research. PMID:24920604

  12. The Rhipicephalus (Boophilus microplus Bm86 gene plays a critical role in the fitness of ticks fed on cattle during acute Babesia bovis infection

    Directory of Open Access Journals (Sweden)

    Knowles Donald P

    2010-11-01

    Full Text Available Abstract Background Rhipicephalus (Boophilus microplus is an economically important tick of cattle involved in the transmission of Babesia bovis, the etiological agent of bovine babesiosis. Commercial anti-tick vaccines based on the R. microplus Bm86 glycoprotein have shown some effect in controlling tick infestation; however their efficacy as a stand-alone solution for tick control has been questioned. Understanding the role of the Bm86 gene product in tick biology is critical to identifying additional methods to utilize Bm86 to reduce R. microplus infestation and babesia transmission. Additionally, the role played by Bm86 in R. microplus fitness during B. bovis infection is unknown. Results Here we describe in two independent experiments that RNA interference-mediated silencing of Bm86 decreased the fitness of R. microplus females fed on cattle during acute B. bovis infection. Notably, Bm86 silencing decreased the number and survival of engorged females, and decreased the weight of egg masses. However, gene silencing had no significant effect on the efficiency of transovarial transmission of B. bovis from surviving female ticks to their larval offspring. The results also show that Bm86 is expressed, in addition to gut cells, in larvae, nymphs, adult males and ovaries of partially engorged adult R. microplus females, and its expression was significantly down-regulated in ovaries of ticks fed on B. bovis-infected cattle. Conclusion The R. microplus Bm86 gene plays a critical role during tick feeding and after repletion during blood digestion in ticks fed on cattle during acute B. bovis infection. Therefore, the data indirectly support the rationale for using Bm86-based vaccines, perhaps in combination with acaricides, to control tick infestation particularly in B. bovis endemic areas.

  13. Epizootiology of Babesia bovis and Babesia bigemina in free-ranging white-tailed deer in northeastern Mexico.

    Science.gov (United States)

    Cantu-C, Antonio; Ortega-S, J Alfonso; García-Vázquez, Zeferino; Mosqueda, Juan; Henke, Scott E; George, John E

    2009-06-01

    Species of Rhipicephalus (Boophilus) ticks are the vectors of babesiosis (cattle fever tick), which are distributed worldwide. White-tailed deer (Odocoileus virginianus) are important secondary hosts for the cattle fever ticks, Rhipicephalus (B.) annulatus and Rhipicephalus (B.) microplus. White-tailed deer are capable of sustaining Boophilus spp. tick populations in the presence or absence of cattle. The objectives of this study were to determine the frequency of Babesia bovis and Babesia bigemina and the prevalence of antibodies to them and identify possible risk factors for bovine babesiosis in white-tailed deer in 3 northeastern states of México. Whole blood and serum samples (n = 457) were collected from white-tailed deer in the states of Coahuila, Nuevo Leon, and Tamaulipas during the spring of 2004. Samples were tested for B. bovis and B. bigemina by nested polymerase chain reaction (n-PCR) (the primers for B. bovis identified the gene Rap-1 and B. bigemina were specific primers) and by an indirect immunofluorescence antibody test (IFAT). A questionnaire was given to each ranch to obtain information about management practices. Logistic regression methods were used to test the association between management factors and the dependent variable of positive n-PCR or IFAT. Nineteen (4.2%) samples were positive to B. bigemina and 6 (1.7%) were positive to B. bovis by n-PCR. Serological testing showed 59.9% (n = 274) of deer sampled were positive to B. bovis and 5.4% (n = 25) were positive to B. bigemina antibodies. The logistic model varied with different dependent variables. With positive n-PCR and B. bigemina as the dependent variable, 3 factors were associated: habitat (presence of brush and exotic grasses; odds ratio (OR), 3.3; 95% confidence interval (CI), 1.3-8.5), grazing system (continuous grazing OR 4.0; CI, 1.3-12.2), and tick treatment frequency (3-4 mo; OR 7.0, CI 1.4-34.3; 5-6 mo; OR, 11.0; CI, 1.9-62.7; > 6 mo; OR, 4.6; CI, 0.9-23.3). These findings

  14. Interesting sequence differences between the pilin gene inversion regions of Moraxella lacunata ATCC 17956 and Moraxella bovis Epp63.

    Science.gov (United States)

    Rozsa, F W; Marrs, C F

    1991-07-01

    The bacterium Moraxella lacunata is a causative agent of human conjunctivitis and keratitis. We have previously reported construction of plasmid pMxL1, which includes a 5.9-kb fragment on which the pilin gene inversion region of M. lacunata resides. The inversion region of pMxL1 was shown to invert when pMxL1 was in an Escherichia coli host cell. In this report, we present Western immunoblot analysis using Moraxella bovis Epp63 anti-I and anti-Q pilin sera which demonstrate that pMxL1 makes pilin only when in orientation 1. The sequence of the pMxL1 plasmid containing the invertible region contains a perfect tandem repeat of 19 bp in the orientation 1 nonexpressed pilin gene at the middle of the recombination junction site. This 19-bp insert causes a frameshift and disrupts the pilin gene. The predicted amino acid sequence of this nonfunctional pilin gene (with the 19-bp repeat subtracted) bears closest resemblance to M. bovis Epp63 Q pilin sequence, although the other (functional) M. lacunata pilin encoded by pMxL1 shows slightly higher homology to Q pilin. Comparison of the pMxL1 sequence with that of the M. bovis Epp63 sequence shows two other particularly interesting differences. One is a 15-bp sequence addition found in pMxL1 at the 60-bp region previously reported as a possible M. bovis recombinational enhancer. The second is an AT deletion in pMxL1 compared with Epp63 within an open reading frame (tfpB) which results in the pMxL1 tfpB open reading frame being one-third shorter than in Epp63. The DNA sequences in these three altered regions from the M. lacunata strain from which pMxL1 was derived were amplified by polymerase chain reaction and sequenced. The parent strain was found to contain the differences seen in pMxL1. Comparison of the M.bovis and M. lacunata pilin gene amino acid sequences is also presented.

  15. Boosting BCG-primed mice with chimeric DNA vaccine HG856A induces potent multifunctional T cell responses and enhanced protection against Mycobacterium tuberculosis.

    Science.gov (United States)

    Ji, Ping; Hu, Zhi-Dong; Kang, Han; Yuan, Qin; Ma, Hui; Wen, Han-Li; Wu, Juan; Li, Zhong-Ming; Lowrie, Douglas B; Fan, Xiao-Yong

    2016-02-01

    The tuberculosis pandemic continues to rampage despite widespread use of the current Bacillus Calmette-Guerin (BCG) vaccine. Because DNA vaccines can elicit effective antigen-specific immune responses, including potent T cell-mediated immunity, they are promising vehicles for antigen delivery. In a prime-boost approach, they can supplement the inadequate anti-TB immunological memory induced by BCG. Based on this, a chimeric DNA vaccine HG856A encoding Mycobacterium tuberculosis (M. tuberculosis) immunodominant antigen Ag85A plus two copies of ESAT-6 was constructed. Potent humoral immune responses, as well as therapeutic effects induced by this DNA vaccine, were observed previously in M. tuberculosis-infected mice. In this study, we further evaluated the antigen-specific T cell immune responses and showed that repeated immunization with HG856A gave modest protection against M. tuberculosis challenge infection and significantly boosted the immune protection primed by BCG vaccination. Enhanced protection was accompanied by increased multifunctional Th1 CD4(+) T cell responses, most notably by an elevated frequency of M. tuberculosis antigen-specific IL-2-producing CD4(+) T cells post-vaccination. These data confirm the potential of chimeric DNA vaccine HG856A as an anti-TB vaccine candidate.

  16. Preparation and stability of agarose microcapsules containing BCG.

    Science.gov (United States)

    Esquisabel, A; Hernandez, R M; Igartua, M; Gascón, A R; Calvo, B; Pedraz, J L

    2002-01-01

    An emulsification/internal gelation method of preparing small-sized agarose microcapsules containing Bacillus Calmette-Guerin (BCG) is reported. Agarose microcapsules have been prepared by the emulsification of the hydrogel within a vegetable oil followed by its gelation due to the cooling of the system. Four different oils (sesame, sweet almonds, camomile and jojoba) were assayed. The rheological analysis of the oils showed a Newtonian behaviour, with viscosity values of 37.7, 51.2, 59.3 and 67.1 mPa s for jojoba, camomile, sesame and sweet almonds oil, respectively. The particle size of the microcapsules obtained ranged from 23.1 microm for the microcapsules prepared with sweet almonds oil to 42.6 microm for those prepared with jojoba. The microcapsule particle size was found to be dependent on the viscosity of the oil used in the emulsification step. The encapsulated BCG was identified by the Difco TB stain set K, followed by observation under optical microscopy. Once prepared, microcapsules were freeze-dried using 5% trehalose as cryoprotectant and the stability of the microcapsules was assayed during 12 months storage at room temperature, observing that agarose microcapsules were stable after 12 months storage, since there was no evidence of alteration in the freeze-dried appearance, resuspension rate, observation under microscope, or particle size.

  17. Bacterial targeted tumour therapy-dawn of a new era.

    Science.gov (United States)

    Wei, Ming Q; Mengesha, Asferd; Good, David; Anné, Jozef

    2008-01-18

    Original observation of patients' spontaneous recovery from advanced tumours after an infection or a "fever" inspired extensive research. As a result, Coley's toxin for the therapy of sarcomas and live Bacillus Calmette-Guerin (BCG) for bladder cancer were born. In addition, three genera of anaerobic bacteria have been shown to specifically and preferentially target solid tumours and cause significant tumour lyses. Initial research had focused on determining the best tumour colonizing bacteria, and assessing the therapeutic efficacy of different strategies either as a single or combination treatment modalities. However, although clinical trials were carried out as early as the 1960s, lack of complete tumour lyses with injection of Clostridial spores had limited their further use. Recent progress in the field has highlighted the rapid development of new tools for genetic manipulation of Clostridia which have otherwise been a hurdle for a long time, such as plasmid transformation using electroporation that bore the problems of inefficiency, instability and plasmid loss. A new Clostridium strain, C. novyi-NT made apathogenic by genetic modification, is under clinical trials. New genetic engineering tools, such as the group II intron has shown promise for genetic manipulation of bacteria and forecast the dawn of a new era for a tumour-targeted bacterial vector system for gene therapy of solid tumours. In this review we will discuss the potential of genetically manipulated bacteria that will usher in the new era of bacterial therapy for solid tumours, and highlight strategies and tools used to improve the bacterial oncolytic capability.

  18. Maternal immune stimulation in mice decreases fetal malformations caused by teratogens.

    Science.gov (United States)

    Holladay, S D; Sharova, L V; Punareewattana, K; Hrubec, T C; Gogal, R M; Prater, M R; Sharov, A A

    2002-02-01

    For unknown reasons, non-specific stimulation of the maternal immune system in pregnant mice has what appears to be a broad-spectrum efficacy for reducing birth defects. Immune stimulation by diverse procedures has proven effective, including footpad injection with Freund's complete adjuvant (FCA), intraperitoneal (IP) injection with inert particles to activate resident macrophages, IP injection with attenuated Bacillus Calmette-Guerin (BCG), and intrauterine injection with allogeneic or zenogeneic lymphocytes. Morphologic lesions that were significantly reduced included cleft palate and associated craniofacial defects, digit and limb defects, tail malformations, and neural tube defect (NTD). Teratogenic stimuli to induce these lesions included chemical agents (2,3,7,8-tetrachlorodibenzo-p-dioxin [TCDD], ethyl carbamate [urethane], methylnitrosourea [MNU], cyclophosphamide [CP], and valproic acid [VA]), physical agents (X-rays, hyperthermia), and streptozocin (STZ)-induced diabetes mellitus. Limited information is available regarding mechanisms by which such immune stimulation reduced fetal dysmorphogenesis. The collective literature suggests the possibility that immunoregulatory cytokines of maternal origin may be the effector molecules in this phenomenon.

  19. Diverse ability of maternal immune stimulation to reduce birth defects in mice exposed to teratogens: a review.

    Science.gov (United States)

    Hrubec, T C; Prater, M R; Mallela, M K; Gogal, R M; Guo, T L; Holladay, S D

    2012-06-01

    Stimulating the maternal immune system before or during pregnancy can dramatically improve morphologic outcome in mice that have been exposed to teratogens. For example, maternal immune stimulation in mice reduced craniofacial and palate defects, heart defects, digit and limb defects, tail malformations and neural tube defects caused by diverse teratogens that included chemical agents, hyperthermia, X-rays and diabetes mellitus. Several different procedures of immune stimulation were effective and included footpad injection with Freund's Complete Adjuvant, intraperitoneal (IP) injection with inert particles or attenuated Bacillus Calmette-Guerin, intrauterine injection with allogenic or xenogenic lymphocytes, or intravascular, intrauterine or IP injection with immunomodulatory cytokines. Limited information is available regarding mechanisms by which such immune stimulation reduces fetal dysmorphogenesis; however, cytokines of maternal origin have been suggested as effector molecules that act on the placenta or fetus to improve development. These collective data raise novel questions about the possibility of unrecognized maternal immune system regulatory activity in normal fetal development. This manuscript reviews the literature showing maternal immune protection against morphologic birth defects. Potential operating mechanisms are discussed, and the possibility is considered that a suppressed maternal immune system may negatively impact fetal development.

  20. Investigation of granulomatous prostatitis incidence following intravesical BCG therapy

    Science.gov (United States)

    Balasar, Mehmet; Doğan, Metin; Kandemir, Abdulkadir; Taskapu, Hakan Hakki; Cicekci, Faruk; Toy, Hatice; Gurbuz, Recai

    2014-01-01

    In the present manuscript, we studied the incidence of granulomatous prostatitis in the prostatectomy specimen of the patients who underwent transurethral resection of the prostate (TURP) after superficial bladder cancer treatment with intravesical Bacillus Calmette-Guerin (BCG) and were diagnosed with benign prostate hyperplasia (BPH). The clinical data and histopathological specimen records of 472 patients who underwent TUR-P due to BPH diagnosis, obtained over a period of 6 years in the urology department of Private Konya Hospital, Konya, Turkey, were studied retrospectively. The cases were divided into two groups as (Group I) who did not undergo any treatment and as (Group II) who underwent BCG treatment. The frequency and the clinical course of the cases with granulomatous prostatitis were studied histopathologically. There were in total 472 patients who underwent TUR-P. Out of the 459 patients who did not undergo BCG treatment (Group I), the histopathological specimen records of 262 (57%) was BPH, of 197 (43%) BPH + chronic prostatitis. Of the second group, 13 cases underwent intravesical BCG treatment before surgical intervention due to superficial bladder CA diagnosis. In this group 4 of the cases were diagnosed as (30%) BPH, 9 as (70%) chronic prostatitis + BPH. 6 out of the 9 chronic prostatitis cases were chronic prostatitis, 2 caseous granulomatous prostatitis, 1 non-caseous granulomatous prostatitis. Granulomatous prostatitis cases should require no specific therapy. Conclusion: In patients with obstruction complaints following intravesical BCG treatment, granulomatous prostatitis should also be considered and treatment plans should be made accordingly. PMID:25035779

  1. Intravesical therapy for urothelial carcinoma of the urinary bladder: a critical review

    Directory of Open Access Journals (Sweden)

    Daher C. Chade

    2009-12-01

    Full Text Available The management of non-muscle-invasive urothelial carcinoma of the bladder (UCB is a challenge for physicians and patients alike. This is largely due to the heterogeneous natural history of this disease, in which tumors range from indolent to rapidly progressive and eventually fatal. Moreover, the high rate of recurrence and progression cause significant morbidity, expense, and detriment to quality of life. The advent of effective and safe intravesical therapies has improved the management of non-muscle-invasive UCB. Nevertheless, despite over 30 years of research and clinical experience, the mechanism, risks, benefits, and optimal regimens and treatment algorithms remain unclear. Although immunotherapy with bacillus Calmette-Guerin (BCG has been the mainstay of intravesical treatment and represents a significant advance in the interaction of immunology and oncology, its clinical effectiveness is accompanied by a wide range of adverse events. Here, we review the literature on intravesical immunotherapy and chemotherapy with the aim of evaluating the clinical utility of the different treatments and providing recommendations. Many studies over the years have compared efficacy and toxicities of different agents and regimens, and certain conclusions are now well supported by high-level evidence. Future perspectives and promising advances in drug development are discussed and areas of improvement are identified in order to promote better cancer control and decrease the rate and severity of side-effects.

  2. A Comparative Study on the Effect of BCG-PSN and Thymopeptides on T-lymphocyte Subsets of Normal and Immunosuppressed Mice

    Institute of Scientific and Technical Information of China (English)

    邓云华; 陈映玲; 陈兴平; 李永喜; 周礼义

    2003-01-01

    To compare the effects of polysaccharide nucleic acid fraction of bacillus calmette guerin (BCG-PSN) and thymopeptides on T-lymphocytes of normal and immunosuppressed mice, CD4+ and CD8+ T-lymphocyte subsets of single nucleic cell in thymus, spleen and peripheral blood were detected successively by flow cytometry after application of BCG-PSN and thymopeptides. Meanwhile, CD4+/CD8+ ratio was also calculated. The results showed that both BCG-PSN and thymopeptides could decrease the proportion of CD4+ CD8+ T-lymphocyte subsets in the thymus, at the same time increase CD4+ T-lymphocyte, CD8+ T-lymphocyte proportion in the three tissues.The fluctuation in amplitude was greater in thymopeptides group than that in BCG-PSN group. It is concluded that acting location of thymopeptides is in thymus, its stimulating action is stronger than that of BCG-PSN, while BCG-PSN not only accelerates the differentiation in thymus, but also has some direct stimulation to peripheral CD4+ T-lymphocytes, and can maintain CD4+/CD8+ ratio within normal range. So, BCG-PSN is safer.

  3. Different effects of BCG strains - A natural experiment evaluating the impact of the Danish and the Russian BCG strains on morbidity and scar formation in Guinea-Bissau

    DEFF Research Database (Denmark)

    Frankel, H; Byberg, S; Andersen, Morten Bjerregaard;

    2016-01-01

    models. Scar prevalence and size were compared using binomial regression and ranksum tests. RESULTS: Among 1206 children, 18% received Danish BCG (n=215) and 82% Russian BCG (n=991). The adjusted hazard ratio (aHR) for consultations was 0.94 (95% CI 0.60-1.46) for Danish BCG compared with Russian BCG......BACKGROUND: Different Bacillus Calmette-Guerin (BCG) vaccine strains may have different non-specific effects. We assessed the effect of two BCG strains (Danish and Russian) on childhood morbidity and BCG scarification in Guinea-Bissau. METHODS: During 2011-2013, infants in the Bandim Health Project....... Girls vaccinated with Danish BCG tended to have lower consultation rates compared with girls vaccinated with Russian BCG (aHR 0.56 (0.25-1.24)), whereas the effect was opposite for boys (aHR 1.24 (0.74-2.11)), p=0.09. Children vaccinated with Danish BCG were more likely to develop a scar (97%) than...

  4. Emerging intravesical therapies for management of nonmuscle invasive bladder cancer

    Directory of Open Access Journals (Sweden)

    Jeffrey J Tomaszewski

    2010-05-01

    Full Text Available Jeffrey J Tomaszewski, Marc C SmaldoneDepartment of Urology, University of Pittsburgh School of Medicine, Pennsylvania, USAAbstract: Transitional cell carcinoma (TCC is the second most common urologic malignancy, and 70% of patients present with superficial or nonmuscle invasive bladder cancer (NMIBC. Intravesical bacillus Calmette-Guerin (BCG is the most effective agent for preventing disease recurrence, and the only therapy able to inhibit disease progression. However, recurrence rates as high as 30% and significant local and systemic toxicity have led to increased interest in alternative intravesical therapies. In patients refractory or intolerant to BCG, BCG-interferon α2b, gemcitabine, and anthracyclines (doxorubicin, epirubicin, valrubicin have demonstrated durable clinical responses. Phase I trials investigating alternative cytotoxic agents, such as apaziquone, taxanes (docetaxel, paclitaxel, and suramin are reporting promising data. Novel immunomodulating agents have demonstrated promise as efficacious alternatives in patients refractory to BCG. Optimization of existing chemotherapeutic regimens using hyperthermia, photodynamic therapy, magnetically-targeted carriers, and liposomes remains an area of active investigation. Despite enthusiasm for new intravesical agents, radical cystectomy remains the treatment of choice for patients with NMIBC who have failed intravesical therapy and selected patients with naïve T1 tumors and aggressive features. This report provides a comprehensive review of contemporary intravesical therapy for NMIBC and refractory NMIBC, with an emphasis on emerging agents and novel treatment modalities.Keywords: transitional cell carcinoma, nonmuscle, invasive, intravesical therapy, BCG

  5. Progress on immunologic mechanism and vaccine development of tuberculosis%结核病的免疫机制及疫苗研发进展

    Institute of Scientific and Technical Information of China (English)

    李萍; 吴利先

    2016-01-01

    Tuberculosis (TB) which is caused by Mycobacterium tuberculosis is a seriously infectious disease that will threaten people's health.Bacillus Calmette-Guerin (BCG) is the only effective vaccine,but it cannot prevent the occurrence and development of TB.Novel vaccines can induce specific immune response and make up for the deficiency of BCG.Meanwhile,the further study of TB mechanism provides a basis and strategy for the development of new drug and vaccine.The recent advances on the immunologic mechanism and vaccine of TB are reviewed in this article.%结核是由结核分枝杆菌引起的严重危害人类健康的重大传染性疾病.卡介苗是目前对结核病唯一有效的疫苗,却不能预防结核病的发生和发展,新型疫苗可特异性诱导机体免疫应答,有助于弥补卡介苗的不足.同时,结核病免疫机制的深入研究为结核治疗药物和新型疫苗的研发提供了依据.本文就结核病的免疫机制及疫苗研发现状作一综述.

  6. REGULAR EXPRESSION OF DISCOIDIN DOMAIN RECEPTOR 2 IN THE IMPROVED ADJUVANT-INDUCED ANIMAL MODEL FOR RHEUMATOID ARTHRITIS

    Institute of Scientific and Technical Information of China (English)

    Wei Li; Yuan-qiang Zhang; Xin-ping Liu; Li-bo Yao; Lan Sun

    2005-01-01

    Objective To investigate the expression of discoidin domain receptor 2 (DDR2) of fibroblast-like synovial cells in improved adjuvant-induced animal (AIA) model for rheumatoid arthritis (RA) and to provide evidence for DDR2′s antagonist use clinically.Methods AIA was modified by administrating 0.1 mL of complete Freund′s adjuvant (CFA, mixed with 5 mg Bacillus Calmette-Guerin vaccine/mL) into rats′ right hind paws and 0.125 mL tumor necrosis factor-α (2 U/mL) into right ankles and subpatellar fatty tissue. The expression of DDR2 in fibroblast-like synovial cells was assessed using immunohistochemistry,immunofluorescence histochemistry, and in situ hybridization methods. Levels of anti-collagen Ⅱ antibody were measured using enzyme-linked immunosorbent assay.Results Given the terms mentioned above, we found a more practical rat model, apparently decreasing immunization time (average 3-5 days). DDR2 can be detected upon the 15th day of immunization; expression gradually increased with time going on, and reaching a peak 35 days after immunization before gradually decreasing. Serum anti-collagen Ⅱ antibody showed similar expression patterns as DDR2, but reached peak later than DDR2, about 40 days after immunization.Conclusion Regular expression of DDR2 in animal models infers its important role in the pathological process of RA.

  7. 结核病疫苗研究进展%Research progress in tuberculosis vaccines

    Institute of Scientific and Technical Information of China (English)

    杨明; 尹文; 汪爱勤

    2010-01-01

    At present, the epidemic of tuberculosis (TB) is severe, and the only available vaccine,Bacillus Calmette-Guerin (BCG), is not high efficacious.Thus, there is a need for the development of novel TB vaccines which are more safe, effective, practical and inexpensive.This review summarizes the advance in the development of TB vaccines, including recombinant BCG vaccine, live attenuated vaccine, subunit vaccine, DNA vaccine and viral vector vaccine.%目前结核病疫情非常严峻,而唯一可用的疫苗卡介苗效果并不理想.因此,需要研发更安全有效和实用价廉的新型疫苗.此文综述了目前正在研发的新型结核病疫苗的研究进展,这些疫苗包括重组卡介苗、减毒活疫苗、亚单位疫苗、DNA疫苗和病毒载体疫苗.

  8. Tuberculosis in Aboriginal Canadians

    Directory of Open Access Journals (Sweden)

    Vernon H Hoeppner

    2000-01-01

    Full Text Available Endemic tuberculosis (TB was almost certainly present in Canadian aboriginal people (aboriginal Canadians denotes status Indians, Inuit, nonstatus Indians and metis as reported by Statistics Canada before the Old World traders arrived. However, the social changes that resulted from contact with these traders created the conditions that converted endemic TB into epidemic TB. The incidence of TB varied inversely with the time interval from this cultural collision, which began on the east coast in the 16th century and ended in the Northern Territories in the 20th century. This relatively recent epidemic explains why the disease is more frequent in aboriginal children than in Canadian-born nonaboriginal people. Treatment plans must account for the socioeconomic conditions and cultural characteristics of the aboriginal people, especially healing models and language. Prevention includes bacillus Calmette-Guerin vaccination and chemoprophylaxis, and must account for community conditions, such as rates of suicide, which have exceeded the rate of TB. The control of TB requires a centralized program with specifically directed funding. It must include a program that works in partnership with aboriginal communities.

  9. Co-infection of tuberculosis and parasitic diseases in humans: a systematic review.

    Science.gov (United States)

    Li, Xin-Xu; Zhou, Xiao-Nong

    2013-03-22

    Co-infection of tuberculosis and parasitic diseases in humans is an important public problem in co-endemic areas in developing countries. However, there is a paucity of studies on co-infection and even fewer reviews. This review examines 44 appropriate papers by PRISMA from 289 papers searched in PubMed via the NCBI Entrez system (no grey literature) up to December 2012 in order to analyze the factors that influence epidemic and host's immunity of co-infection. The limited evidence in this review indicates that most common parasite species are concurrent with Mycobacterium tuberculosis in multiple organs; socio-demographics such as gender and age, special populations with susceptibility such as renal transplant recipients, patients on maintenance haemodialysis, HIV positive patients and migrants, and living in or coming from co-endemic areas are all likely to have an impact on co-infection. Pulmonary tuberculosis and parasitic diseases were shown to be risk factors for each other. Co-infection may significantly inhibit the host's immune system, increase antibacterial therapy intolerance and be detrimental to the prognosis of the disease; in addition, infection with parasitic diseases can alter the protective immune response to Bacillus Calmette-Guerin vaccination against Mycobacterium tuberculosis.

  10. 疫苗相关麻痹型脊灰病例死亡1例

    Institute of Scientific and Technical Information of China (English)

    郝振忠

    2011-01-01

    1 病例摘要 患儿,男,2006-05-02出生,第一胎,剖腹产,发育及体重正常.父母体健,否认家族疾病史.该患儿属于本地常住儿童,生前无外出史.该儿童于出生当日接种乙型肝炎疫苗(Hepatitis B Vaccine,HepB)第1针;2006-06-19接种HepB第2针、卡介苗(Bacillus Calmette Guerin,BCG)1针次;2006-07-20口服脊髓灰质炎减毒活疫苗(Oral Poliomyelitis Attenuated Live Vaccine,OPV)第1剂,2006-08-20接种OPV 第2剂(北京天坛生物制品股份有限公司生产,批号2005111111,有效期至2007年11月)、接种百日咳-白喉-破伤风联合疫苗(Diphtheria,Tetanus,Pertussis Combined Vaccine;DTP)第1针. 该患儿于2006-08-20接种OPV第2剂,半月后于2006-09-05出现发热(当时体温38℃)、恶心、呕吐、腹泻,双下肢无力.

  11. BCG LYMPHADENITIS

    Directory of Open Access Journals (Sweden)

    Vijayakumar

    2014-08-01

    Full Text Available Background: The World Health Organization (WHO has recommended Bacillus Calmette-Guerin (BCG vaccination as a part of the global expanded program for immunization. Although the BCG vaccine is usually a safe vaccine, a number of complications with lymphadenitis being the most common complication, can occur. AIM: The aim of the present study was to evaluate the clinical presentation and the histomorphological features of BCG adenitis in children. RESULTS: A total of 60 patients with BCG lymphadenitis presented between June 2010 and December 2013. The most common age of presentation was 3 months. In the majority (50 of the cases, the lymphadenitis involved ipsilateral left axillary nodes. Other sites of involvement included the left supraclavicular lymph nodes in 5 (8.3% patients, and both the left axillary and supraclavicular lymph nodes were involved in 5 cases (8.3%. All the patients had history of BCG vaccination prior to the onset of lymphadenitis. CONCLUSION: Diagnosis of BCG lymphadenitis is clinical. Parental education and awareness among paramedical personnel, including general practitioners, is essential so that prompt recognition and management of BCG adenitis can be ensured.

  12. Advance on vaccine against toxoplasmosis%弓形虫病疫苗研究进展概述

    Institute of Scientific and Technical Information of China (English)

    李运娜; 黄金贵; 张西臣

    2011-01-01

    弓形虫是一种专性细胞内寄生原虫,能引起人畜共患弓形虫病,严重威胁着人类健康且对畜牧业的发展造成巨大的经济损失.研制安全有效的疫苗是防制弓形虫病的策略之一.目前,弓形虫病疫苗包括灭活疫苗、减毒活疫苗、核酸疫苗、亚单位疫苗和卡介苗.本文对弓形虫病疫苗的研究现状进行了综述.%Toxoplasma gondii is an obligate intracellular parasite, which is capable of infecting a variety of animal and human, causing toxoplasmosis.It has important effects on human’s health and can cause considerable economic losses to the development of animal husbandry.One of the most effective prevention and treatment strategies to toxoplasmosis is to develop safe and effective vaccines.The vaccine of toxoplasmosis include inactivated vaccine, live attenuated vaccine, subunit vaccine, the nucleic acid vaccine and Bacillus Calmette-Guerin.This paper reviewed the research of toxoplasmosis vaccine in recent years.

  13. Comparison of the fibronectin-binding ability and antitumor efficacy of various mycobacteria.

    Science.gov (United States)

    Hudson, M A; Ritchey, J K; Catalona, W J; Brown, E J; Ratliff, T L

    1990-07-01

    Although the mechanism by which Bacillus Calmette-Guerin (BCG) exerts an antitumor effect on superficial bladder tumors is not fully understood, recent evidence has implicated binding of BCG organisms to fibronectin (FN) as requisite for this antitumor efficacy. Various substrains of BCG and other mycobacteria were tested in vitro for their relative capacities to bind both matrix and soluble FN. A substrain of Mycobacterium kansasii, designated the "high-binding strain," was found to bind FN more readily (P less than 0.05) in in vitro studies, when compared to commercially available substrains of BCG (Tice, Connaught, and Armand Frappier). The binding by the three commercial strains of BCG to FN in vitro appeared to be equivalent. The high-binding strain was further demonstrated to attach more readily in vivo to the acutely injured murine bladder (P less than 0.005) than the Armand Frappier substrain. Finally, using the MB49 murine bladder tumor model, an enhanced antitumor effect (P less than 0.05) was noted in mice treated with intravesical high-binding strain, in comparison to the Armand Frappier substrain, during five weekly treatments. It appears not only that the commercial substrains of BCG bind FN in an equivalent manner but also that the relative binding capacities of the substrains correlate directly with antitumor activity. A substrain of M. kansasii appears to have been identified which may prove more clinically effective than the currently available strains of BCG.

  14. Role of fibronectin in intravesical BCG therapy for superficial bladder cancer.

    Science.gov (United States)

    Ratliff, T L; Kavoussi, L R; Catalona, W J

    1988-02-01

    Intravesical bacillus Calmette-Guerin (BCG) has been demonstrated to be effective both for prophylaxis and treatment of superficial bladder cancer. In order to identify the progression of events that result in BCG-mediated antitumor activity, studies were performed to evaluate the mechanism of binding of BCG within the bladder. Histological and quantitative studies in a mouse model revealed that BCG attached to the bladder wall only in areas of urothelial damage. Preliminary in vitro data showed that BCG attached to surfaces coated with extracellular matrix proteins. Further studies were then performed using purified extracellular matrix proteins to identify the proteins responsible for attachment. BCG were observed to attach to surfaces coated only with purified fibronectin (FN) but not to other purified proteins including laminin, collagen or fibrinogen. The attachment of BCG to purified FN in vitro was dose dependent and was inhibited by anti-FN antibodies. Moreover, BCG attachment in vivo to bladders with damaged urothelial surfaces was inhibited more than 95% by anti-FN antibodies, but binding was not affected by anti-laminin antibodies or preimmune serum. A survey of commercially available BCG vaccines (Pasteur, Tice, Glaxo, Connaught) showed that only Glaxo BCG did not attach to FN-coated surfaces. Glaxo BCG also was shown to express inferior antitumor activity suggesting that the absence of FN binding by Glaxo may have been associated with the absence of antitumor activity of the vaccine.

  15. Effects of tumor necrosis factor-alpha and interferon-gamma on expressions of matrix metalloproteinase-2 and -9 in human bladder cancer cells.

    Science.gov (United States)

    Shin, K Y; Moon, H S; Park, H Y; Lee, T Y; Woo, Y N; Kim, H J; Lee, S J; Kong, G

    2000-10-31

    We have investigated the effects of tumor necrosis factor-alpha (TNF-alpha) and interferon (INF-gamma), the potent Bacillus Calmette-Guerin (BCG)-induced cytokines on the production of MMP-2, MMP-9, TIMP-1, TIMP-2 and MT1-MMP in high grade human bladder cancer cell lines, T-24, J-82 and HT-1376 cell lines. MMP-2 expression and activity were decreased in T-24 cells treated with both cytokines in a dose dependent manner. However, J-82 cells treated with TNF-alpha and INF-gamma revealed dose dependent increases of MMP-9 expression and activity with similar baseline expression and activity of MMP-2. HT-1376 cells after exposure to TNF-alpha only enhanced the expression and activity of MMP-9. These results indicate that TNF-alpha and INF-gamma could regulate the production of MMP-2 or MMP-9 on bladder cancer cells and their patterns of regulation are cell specific. Furthermore, this diverse response of bladder cancer cells to TNF-alpha and INF-gamma suggests that BCG immunotherapy may enhance the invasiveness of bladder cancer in certain conditions with induction of MMPs.

  16. [Assessment of BCG vaccine practices].

    Science.gov (United States)

    Lechiche, C; Charpille, M; Saissi, G; Sotto, A

    2016-01-01

    Tuberculosis is a major public health problem. In France, the vaccine against tuberculosis (Bacillus Calmette-Guerin, BCG) is in decline. This decline is firstly due to changes in BGG administration that were implemented in 2006 and secondly because of new recommandations in 2007 that ended compulsory vaccination. To determine their position on this vaccine, in 2013-2014 we asked general practitioners, pediatricians, and Maternal and Infantile Protection Center physicians in the Gard and Herault departments (in Southern France) why this vaccine was not administered and their suggestions for improvement. Most of these doctors (73.9%) stated that they did not oppose this vaccination for children. They expressed concern about potential side effects, technical problems (intradermic injection, multi-dose bottles) and parents' refusal. One quarter of these physicians would have preferred that this vaccine remains compulsory and one third that this vaccine be administered in the maternity hospital. They also requested simplified criteria for patient eligibility, technical improvements (training for intradermal injection, single-dose vaccine) and more information for the public concerning this vaccination.

  17. Intravesical electro-osmotic administration of mitomycin C.

    Science.gov (United States)

    Di Stasi, Savino M; Verri, Cristian; Celestino, Francesco; De Carlo, Francesco; Pagliarulo, Vincenzo

    2016-10-04

    Bladder cancer is very common and most cases are diagnosed as nonmuscle invasive disease, which is characterized by its propensity to recur and progress. Intravesical therapy is used to delay recurrence and progression, while cystectomy is reserved for patients who are refractory to transurethral resection and intravesical therapy. There is an increasing interest in methods to enhance the delivery of intravesical chemotherapeutic agents to improve efficacy. In vitro and in vivo studies demonstrated that electro-osmosis of mitomycin C (MMC) is more effective in delivering this drug into the urothelium, lamina propria, and superficial muscle layers of the bladder wall than is passive transport. Higher MMC tissue concentrations might have a clinical impact in the treatment of nonmuscle invasive bladder cancer (NMIBC). In randomized trials, intravesical electro-osmotic MMC was associated with superior response rate in high-risk NMIBC cancer, compared with passive diffusion MMC transport. New strategies such as intravesical Bacillus Calmette-Guerin (BCG) combined with electro-osmotic MMC as well as intravesical pre-operative electro-osmotic MMC provided promising results in terms of higher remission rates and longer remission times.Device-assisted intravesical chemotherapy may be a useful ancillary procedure in the treatment of NMIBC. Its evaluation must be planned with respect to the technical functioning of equipment and their use for a clear purpose to avoid the financial and human costs associated with incorrect therapies.

  18. Effect of Early Intervention Applied to Patients with Chronic Obstructive Pulmonary Disease at Different Stages

    Directory of Open Access Journals (Sweden)

    W Lei

    2014-04-01

    Full Text Available Objective: Early intervention in chronic obstructive pulmonary disease (COPD includes health education, smoking cessation, pulmonary rehabilitation and enhancing immunity (administration of influenza vaccine and polysaccharide nucleic acid fraction of bacillus Calmette-Guerin [BCG-PSN]. The effect of early intervention was investigated systematically in patients with COPD at different stages. Methods: We enrolled 422 patients with COPD at different stages without symptoms and then randomly assigned them to intervention and control groups. The intervention group was provided with early intervention and usual care while the control group was only provided with usual care. One year of follow-up was performed to observe forced expiratory volume in one second (FEV1, FEV1/forced vital capacity (FVC, and the ratio of patients with acute exacerbation (number of patients with acute exacerbation/total of patients. Results: The values of decline in FEV1 and FEV1/FVC were significantly lower in the intervention groups of stage I and II than control groups (all p 0.05. The ratios of patients with acute exacerbation were lower in the intervention groups of all the stages than the control groups (p < 0.05. Conclusion: Early intervention could slow the decline of FEV1 and FEV1/FVC in patients with COPD in stages I and II, but not in stages III and IV. Early intervention could also prevent patients with COPD from getting acute exacerbation and improve their quality of life in all the stages of the disease.

  19. Construction of two Listeria ivanovii attenuated strains expressing Mycobacterium tuberculosis antigens for TB vaccine purposes.

    Science.gov (United States)

    Lin, Qingqing; Zhou, Mengying; Xu, Zongkai; Khanniche, Asma; Shen, Hao; Wang, Chuan

    2015-02-20

    Bacillus Calmette-Guerin (BCG) has failed in complete control of tuberculosis (TB), thus, novel tuberculosis vaccines are urgently needed. We have constructed several TB vaccine candidates, which are characterized by the use of Listeria ivanovii (LI) strain as an antigen delivery vector. Two L. ivanovii attenuated recombinant strains L. ivanovii△actAplcB-Rv0129c and L. ivanovii△actAplcB-Rv3875 were successfully screened. Results from genome PCR and sequencing showed that the Mycobacterium tuberculosis antigen gene cassette coding for Ag85C or ESAT-6 protein respectively had been integrated into LI genome downstream of mpl gene. Western blot confirmed the secretion of Ag85C or ESAT-6 protein from the recombinant LI strains. These two recombinant strains showed similar growth curves as wide type strain in vitro. In vivo, they transiently propagated in mice spleen and liver, and induced specific CD8(+) IFN-γ secretion. Therefore, in this paper, two novel LI attenuated strains expressing specific TB antigens were successfully constructed. The promising growth characteristics in mice immune system and the capability of induction of IFN-γ secretion make them of potential interest for development of TB vaccines.

  20. In vitro culture of Babesia bovis in a bovine serum-free culture medium supplemented with insulin, transferrin, and selenite.

    Science.gov (United States)

    Rojas Martínez, C; Rodríguez-Vivas, R I; Figueroa Millán, J V; Acosta Viana, K Y; Gutiérrez Ruiz, E J; Álvarez Martínez, J A

    2016-11-01

    Bovine serum is an important factor for the optimal growth of Babesia bovis in vitro. This protozoan can be cultured in M-199 with Earle's salts medium (M-199) supplemented with 40% bovine serum (BS). In the present study, four media were assessed along with the control medium M-199. The effect on the proliferation of B. bovis in vitro was tested when these media were combined with insulin (Ins), transferrin (Trans) and selenite (Sel) in the absence of bovine serum. Treatment with Advanced DMEM/F12 medium (A-DMEM/F12) achieved the highest percentage of parasitized erythrocytes (PPE), reaching a maximum value of 9.59%. A-DMEM/F12 medium supplemented with a mixture of Ins (2000 mg/L), Trans (1100 mg/L), and Sel (1.34 mg/L) allowed for the adaptation and proliferation of B. bovis without bovine serum, showed a constant increase in PPE, and reached a maximum value of 9.7% during seven cycles of in vitro culture. It was concluded that continuous proliferation of B. bovis in vitro could be achieved using A-DMEM/F12 medium supplemented with Ins-Trans-Sel, without bovine serum. After adaptation for proliferation in serum-free medium, the B. bovis strain of parasites could have future use in the study of this economically important protozoan species that affects cattle.

  1. Detection of Babesia bovis in blood samples and its effect on the hematological and serum biochemical profile in large ruminants from Southern Punjab

    Institute of Scientific and Technical Information of China (English)

    Samreen Zulfiqar; Ali Saeed; Furhan Iqbal; Sadia Shahnawaz; Muhammad Ali; Arif Mahmood Bhutta; Shahid Iqbal; Sikandar Hayat; Shazia Qadir; Muhammad Latif; Nazia Kiran

    2012-01-01

    Objective:To determine the presence of Babesia bovis (B. bovis) in large ruminants in southern Punjab and its effect on hematological and serum biochemical profile of host animals. Methods:Blood samples were collected from 144 large ruminants, including 105 cattle and 39 buffaloes, from six districts in southern Punjab including Multan, Layyah, Muzaffar Garh, Bhakar, Bahawalnagar and Vehari. Data on the characteristics of animals and herds were collected through questionnaires. Different blood (hemoglobin, glucose) and serum (ALT, AST, LDH, cholesterol) parameters of calves and cattle were measured and compared between parasite positive and negative samples to demonstrate the effect of B. bovis on the blood and serological profile of infected animals. Results:27 out of 144 animals, from 5 out of 6 sampling districts, produced the 541-bp fragment specific for B. bovis. Age of animals (P=0.02), presence of ticks on animals (P=0.04) and presence of ticks on dogs associated with herds (P=0.5) were among the major risk factors involved in the spread of bovine babesiosis in the study area. ALT concentrations were the only serum biochemical values that significantly varied between parasite positive and negative cattle. Conclusions:This study has reported for the first time the presence of B. bovis in large ruminant and the results can lead to the prevention of babesiosis in the region to increase the livestock output.

  2. Phylogenetic relationships of Mongolian Babesia bovis isolates based on the merozoite surface antigen (MSA)-1, MSA-2b, and MSA-2c genes.

    Science.gov (United States)

    Altangerel, Khukhuu; Sivakumar, Thillaiampalam; Battsetseg, Badgar; Battur, Banzragch; Ueno, Akio; Igarashi, Ikuo; Yokoyama, Naoaki

    2012-03-23

    We conducted a molecular epidemiological study on Babesia bovis in Mongolia. Three hundred blood samples collected from cattle grazed in seven different districts were initially screened using a previously established diagnostic polymerase chain reaction (PCR) assay for the detection of B. bovis-specific DNA. Positive samples were then used to amplify and sequence the hyper-variable regions of three B. bovis genes encoding the merozoite surface antigen (MSA)-1, MSA-2b, and MSA-2c. The diagnostic PCR assay detected B. bovis among cattle populations of all districts surveyed (4.4-26.0%). Sequences of each of the three genes were highly homologous among the Mongolian isolates, and found in a single phylogenetic cluster. In particular, a separate branch was formed only by the Mongolian isolates in the MSA-2b gene-based phylogenetic tree. Our findings indicate that effective preventative and control strategies are essential to control B. bovis infection in Mongolian cattle populations, and suggest that a careful approach must be adopted when using immunization techniques.

  3. Auxotrophic complementation as a selectable marker for stable expression of foreign antigens in Mycobacterium bovis BCG.

    Science.gov (United States)

    Borsuk, Sibele; Mendum, Tom A; Fagundes, Michel Quevedo; Michelon, Marcelo; Cunha, Cristina Wetzel; McFadden, Johnjoe; Dellagostin, Odir Antônio

    2007-11-01

    Mycobacterium bovis BCG has the potential to be an effective live vector for multivalent vaccines. However, most mycobacterial cloning vectors rely on antibiotic resistance genes as selectable markers, which would be undesirable in any practical vaccine. Here we report the use of auxotrophic complementation as a selectable marker that would be suitable for use in a recombinant vaccine. A BCG auxotrophic for the amino acid leucine was constructed by knocking out the leuD gene by unmarked homologous recombination. Expression of leuD on a plasmid not only allowed complementation, but also acted as a selectable marker. Removal of the kanamycin resistance gene, which remained necessary for plasmid manipulations in Escherichia coli, was accomplished by two different methods: restriction enzyme digestion followed by re-ligation before BCG transformation, or by Cre-loxP in vitro recombination mediated by the bacteriophage P1 Cre Recombinase. Stability of the plasmid was evaluated during in vitro and in vivo growth of the recombinant BCG in comparison to selection by antibiotic resistance. The new system was highly stable even during in vivo growth, as the selective pressure is maintained, whereas the conventional vector was unstable in the absence of selective pressure. This new system will now allow the construction of potential recombinante vaccine strains using stable multicopy plasmid vectors without the inclusion of antibiotic resistance markers.

  4. Sequence analysis of the inversion region containing the pilin genes of Moraxella bovis.

    Science.gov (United States)

    Fulks, K A; Marrs, C F; Stevens, S P; Green, M R

    1990-01-01

    Moraxella bovis EPP63 is able to produce two antigenically distinct pili called Q and I pili (previously called beta and alpha pili). Hybridization studies have shown that the transition between the types is due to inversion of a 2.1-kilobase segment of chromosomal DNA. We present the sequence of a 4.1-kilobase region of cloned DNA spanning the entire inversion region in orientation 1 (Q pilin expressed). Comparison of this sequence with the sequence of the polymerase chain reaction-amplified genomic DNA from orientation 2 (I pilin expressed) allows the site-specific region of recombination to be localized to a 26-base-pair region in which sequence similarity to the left inverted repeat of the Salmonella typhimurium hin system was previously noted. In addition, 50% sequence similarity was seen in a 60-base-pair segment of our sequence to the recombinational enhancer of bacteriophage P1, an inversion system related to the hin system of S. typhimurium. Finally, two open reading frames representing potential genes were identified.

  5. Estimating the power of a Mycobacterium bovis vaccine trial in Irish badgers.

    Science.gov (United States)

    Aznar, I; More, S J; Frankena, K; De Jong, M C M

    2013-09-01

    The aim of this study was to estimate the power, using simulation techniques, of a group randomized vaccine field trial designed to assess the effect of vaccination on Mycobacterium bovis transmission in badgers. The effects of sample size (recapture percentage), initial prevalence, sensitivity and specificity of the diagnostic test, transmission rate between unvaccinated badgers, Vaccine Efficacy for Susceptibility (VES) and Vaccine Efficacy for Infectiousness (VEI), on study power were determined. Sample size had a small effect on power. Study power increased with increasing transmission rate between non-vaccinated badgers. Changes in VES had a higher impact on power than changes in VEI. However, the largest effect on study power was associated with changes in the specificity of the diagnostic test, within the range of input values that were used for all other modelled parameters. Specificity values below 99.4% yielded a study power below 50% even when sensitivity was 100% and, VEI and VES were both equal to 80%. The effect of changes in sensitivity on study power was much lower. The results from our study are in line with previous studies, as study power was dependent not only on sample size but on many other variables. In this study, additional variables were studied, i.e. test sensitivity and specificity. In the current vaccine trial, power was highly dependent on the specificity of the diagnostic test. Therefore, it is critical that the diagnostic test used in the badger vaccine trial is optimized to maximize test specificity.

  6. Expression, purification and crystallization of an atypical class C acid phosphatase from Mycoplasma bovis.

    Science.gov (United States)

    Singh, Harkewal; Reilly, Thomas J; Calcutt, Michael J; Tanner, John J

    2011-10-01

    Class C acid phosphatases (CCAPs) are 25-30 kDa bacterial surface proteins that are thought to function as broad-specificity 5',3'-nucleotidases. Analysis of the newly published complete genome sequence of Mycoplasma bovis PG45 revealed a putative CCAP with a molecular weight of 49.9 kDa. The expression, purification and crystallization of this new family member are described here. Standard purification procedures involving immobilized metal-ion affinity chromatography and ion-exchange chromatography yielded highly pure and crystallizable protein. Crystals were grown in sitting drops at room temperature in the presence of PEG 3350 and HEPES buffer pH 7.5 and diffracted to 2.3 Å resolution. Analysis of diffraction data suggested a primitive monoclinic space group, with unit-cell parameters a = 78, b = 101, c = 180 Å, β = 92°. The asymmetric unit is predicted to contain six molecules, which are likely to be arranged as three dimers.

  7. LIMITED ANTIBODY EVIDENCE OF EXPOSURE TO MYCOBACTERIUM BOVIS IN FERAL SWINE (SUS SCROFA) IN THE USA.

    Science.gov (United States)

    Pedersen, Kerri; Miller, Ryan S; Anderson, Theodore D; Pabilonia, Kristy L; Lewis, Jonathan R; Mihalco, Rebecca L; Gortázar, Christian; Gidlewski, Thomas

    2017-01-01

    Bovine tuberculosis is a chronic disease of cattle ( Bos taurus ) caused by the bacterium Mycobacterium bovis . Efforts have been made in the US to eradicate the disease in cattle, but spillover into wildlife and subsequent spillback have impeded progress in some states. In particular, infection in white-tailed deer ( Odocoileus virginianus ) has been followed by infection in cattle in some Midwestern states. Infection has also been documented in feral swine ( Sus scrofa ) on the Hawaiian island of Molokai and in various European countries, but no large-scale survey of antibody exposure to the bacteria has been conducted in feral swine in the US. We tested 488 sera from feral swine collected near previously documented outbreaks of bovine tuberculosis in cattle and captive cervids, in addition to 2,237 feral swine sera collected across the US from 1 October 2013 to 30 September 2014. While all but one of the samples were antibody negative, the results are important for establishing baseline negative data since feral swine are capable reservoirs and could be implicated in future outbreaks of the disease.

  8. Pathology and diagnosis of Mycobacterium bovis in naturally infected dromedary camels (Camelus dromedarius) in India.

    Science.gov (United States)

    Narnaware, Shirish Dadarao; Dahiya, Shyam Singh; Tuteja, Fateh Chand; Nagarajan, Govindasamy; Nath, Kashi; Patil, Nitin Vasantrao

    2015-12-01

    The present study investigated the pathological features of tuberculosis (TB) caused by Mycobacterium bovis and its diagnosis in naturally infected dromedary camels from an organized farm in India. During the period of the 5-year study, a total of 18 (19.56 %) camels out of 92 examined showed gross lesions compatible with TB at post-mortem. The clinical signs and pathological lesions in these camels were studied, and the efficacy of different diagnostic tests was also assessed. On the basis of occurrence and distribution of gross TB lesions, the infected camels revealed two different lesional patterns as pulmonary (n = 15) and disseminated (n = 3) form. The histopathology of affected organs revealed typical granulomatous lesions wherein the giant cells and acid-fast bacilli were occasionally observed in pulmonary form whereas they frequently observed in disseminated form. The single intradermal tuberculin test (SIDT) detected TB in 10 (55.55 %) whereas the Ziehl-Neelsen (ZN) stain and IS6110 PCR from tissue lesions detected 13 (72.22 %) and 18 (100 %) of the infected camels, respectively. The study suggests that pulmonary form of the TB is more common in camels indicating respiratory route as the major source of exposure in camel herds. Moreover, very low sensitivity of SIDT was observed which highlights the difficulty for confirmation of TB in live camels.

  9. Protective Effect of Calculus Bovis Sativus on Dextran Sulphate Sodium-Induced Ulcerative Colitis in Mice

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    Xiping Li

    2015-01-01

    Full Text Available Calculus Bovis Sativus (CBS is a commonly used traditional Chinese medicine, which has been reported to exhibit antispasmodic, fever-reducing, anti-inflammatory, and gallbladder-repairing effects. The present study aims to investigate the protective effect of CBS on dextran sulphate sodium- (DSS- induced ulcerative colitis (UC in mice. C57BL/6 male mice were exposed to 5% DSS in drinking water. CBS was given orally at 50 and 150 mg/kg once per day for 7 days. Body weight, disease activity index (DAI, colon length, colonic myeloperoxidase (MPO activity, superoxide dismutase (SOD activity, and malondialdehyde (MDA and nitric oxide (NO levels were measured. Administration of CBS significantly reserved these changes, decreased the MPO activity and MDA and NO level, and increased the SOD activity in the colon tissue. Histological observation suggested that CBS alleviated edema, mucosal damage, and inflammatory cells infiltration induced by DSS in the colon. Moreover, CBS significantly downregulated the mRNA expression of tumor necrosis factor-α (TNF-α, interleukin- (IL- 1β and IL-6 in the colon tissue. Our data suggested that CBS exerted protective effect on DSS-induced UC partially through the antioxidant and anti-inflammatory activities.

  10. Mycobacterium leprae phenolglycolipid-1 expressed by engineered M. bovis BCG modulates early interaction with human phagocytes.

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    Guillaume Tabouret

    Full Text Available The species-specific phenolic glycolipid 1 (PGL-1 is suspected to play a critical role in the pathogenesis of leprosy, a chronic disease of the skin and peripheral nerves caused by Mycobacterium leprae. Based on studies using the purified compound, PGL-1 was proposed to mediate the tropism of M. leprae for the nervous system and to modulate host immune responses. However, deciphering the biological function of this glycolipid has been hampered by the inability to grow M. leprae in vitro and to genetically engineer this bacterium. Here, we identified the M. leprae genes required for the biosynthesis of the species-specific saccharidic domain of PGL-1 and reprogrammed seven enzymatic steps in M. bovis BCG to make it synthesize and display PGL-1 in the context of an M. leprae-like cell envelope. This recombinant strain provides us with a unique tool to address the key questions of the contribution of PGL-1 in the infection process and to study the underlying molecular mechanisms. We found that PGL-1 production endowed recombinant BCG with an increased capacity to exploit complement receptor 3 (CR3 for efficient invasion of human macrophages and evasion of inflammatory responses. PGL-1 production also promoted bacterial uptake by human dendritic cells and dampened their infection-induced maturation. Our results therefore suggest that M. leprae produces PGL-1 for immune-silent invasion of host phagocytic cells.

  11. Bacillus cereus endocarditis in native aortic valve.

    Science.gov (United States)

    Ngow, H A; Wan Khairina, W M N

    2013-02-01

    Bacillus cereus endocarditis is rare. It has been implicated in immunocompromised individuals, especially in intravenous drug users as well as in those with a cardiac prosthesis. The patient was a 31-year-old ex-intravenous drug addict with a past history of staphylococcal pulmonary valve endocarditis, who presented with symptoms of decompensated cardiac failure. Echocardiography showed severe aortic regurgitation with an oscillating vegetation seen on the right coronary cusp of the aortic valve. The blood cultures grew Bacillus cereus. We report this as a rare case of Bacillus cereus endocarditis affecting a native aortic valve.

  12. Avaliação do tratamento com isoniazida em bovinos infectados naturalmente pelo Mycobacterium bovis Evaluation of the treatment with isoniazid carried out in bovines naturally infected with Mycobacterium bovis

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    P.M.P.C. Mota

    2004-10-01

    Full Text Available Foi adotado o tratamento intermitente da tuberculose com isoniazida em 240 bovinos de um rebanho naturalmente infectado pelo M. bovis. No início do tratamento 36,6% dos animais foram reagentes positivos e 2,9% inconclusivos. Todos os animais do rebanho receberam isoniazida por via oral em doses de 25mg/kg de peso vivo, administrada três vezes por semana, às segundas, quartas e sextas-feiras, durante 10 meses, perfazendo 120 doses. A cura de 98,9% dos animais tratados foi verificada por meio da dessensibilização alérgica realizada pela tuberculinização e controle bacteriológico de 39 animais abatidos. O tratamento neste rebanho não resultou em seleção de cepas resistentes a isoniazida, comprovada pelo teste de sensibilidade a essa droga, realizado em cultura de M. bovis isolada de um animal não curado.The intermittent treatment with isoniazid was adopted in 240 bovines from a naturally infected herd. At the beginning of the treatment, 36.6% of the animals were positive reactors and 2.9% inconclusive reactors. The drug was administered orally at a dose of 25mg/kg of live weight, three times a week (on Mondays, Wednesdays and Fridays during 10 months in a total of 120 doses. The cure of 98.9% of the treated animals was verified by the allergic desensitization demonstrated through the tuberculin test and the bacteriological control of 39 slaughtered animals. The treatment in this herd did not cause selection of isoniazid-resistant strains as demonstrated by the isoniazid sensitivity test carried out in M. bovis culture isolated from a non-cured animal.

  13. Assessment of an oral Mycobacterium bovis BCG vaccine and an inactivated M. bovis preparation for wild boar in terms of adverse reactions, vaccine strain survival, and uptake by nontarget species.

    Science.gov (United States)

    Beltrán-Beck, Beatriz; Romero, Beatriz; Sevilla, Iker A; Barasona, Jose A; Garrido, Joseba M; González-Barrio, David; Díez-Delgado, Iratxe; Minguijón, Esmeralda; Casal, Carmen; Vicente, Joaquín; Gortázar, Christian; Aranaz, Alicia

    2014-01-01

    Wildlife vaccination is increasingly being considered as an option for tuberculosis control. We combined data from laboratory trials and an ongoing field trial to assess the risk of an oral Mycobacterium bovis BCG vaccine and a prototype heat-inactivated Mycobacterium bovis preparation for Eurasian wild boar (Sus scrofa). We studied adverse reactions, BCG survival, BCG excretion, and bait uptake by nontarget species. No adverse reactions were observed after administration of BCG (n = 27) or inactivated M. bovis (n = 21). BCG was not found at necropsy (175 to 300 days postvaccination [n = 27]). No BCG excretion was detected in fecal samples (n = 162) or in urine or nasal, oral, or fecal swab samples at 258 days postvaccination (n = 29). In the field, we found no evidence of loss of BCG viability in baits collected after 36 h (temperature range, 11°C to 41°C). Camera trapping showed that wild boar (39%) and birds (56%) were the most frequent visitors to bait stations (selective feeders). Wild boar activity patterns were nocturnal, while diurnal activities were recorded for all bird species. We found large proportions of chewed capsules (29%) (likely ingestion of the vaccine) and lost baits (39%) (presumably consumed), and the proportion of chewed capsules showed a positive correlation with the presence of wild boar. Both results suggest proper bait consumption (68%). These results indicate that BCG vaccination in wild boar is safe and that, while bait consumption by other species is possible, this can be minimized by using selective cages and strict timing of bait deployment.

  14. Protein Spesifik Cairan Kista Cysticercus bovis pada Sapi Bali yang Diinfeksi dengan Taenia saginata (SPECIFIC PROTEIN OF CYSTICERCUS BOVIS CYST FLUID ON BALI CATTLE EXPERIMENTALLY INFECTED WITH TAENIA SAGINATA

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    Nyoman Sadra Dharmawan

    2013-08-01

    Full Text Available Cysticercus bovis is the larval stage of Taenia saginata, the bovine tapeworm. The infection of thislarval in cattle musculature causes Bovine cysticercosis or Cysticercosis bovis.  Bovine cysticercosis is foundworldwide, but mostly in developing countries, where unhygienic conditions, poor cattle managementpractices, and the absence of meat inspection are common.  The adult Taenia infection in man is referredto as taeniasis.  Taenia saginata taeniasis is also found almost all over the world.  The prevalence ofTaenia saginata taeniasis has reported up to 27.5% in Gianyar Bali. In order to control the diseases,vaccination against the larvae stages in cattle of Taenia saginata may play an important role in controllingthe disease in the endemic regions.  The aims of the present study were to prepare and to investigate theimmunogenic protein as vaccine candidate for controlling  Cysticercus bovis infection in in Bali cattle.Cysticercus protein from the cyst fluid was firstly used to immunize mice and the mice sera were thencollected. Cysticercus proteins then analyzed using sodium dodecyl sulfate-gel electrophoresis (SDS-PAGE.All cysticercus proteins were then visualized by Commasie blue staining. The proteins were also transferredonto nitrocellulose membrane and the immunogenic proteins were visualized by Western Blotting usingimmune sera raised in mice.  By Commasie blue staining, a total of 17 proteins were detected with themolecular weight of 14,86 kDa -122,40 kDa from the smallest to the largest. As many as 7 immunogenicproteins with the molecular weights of 16.81 kDa; 19.22 kDa; 20.98 kDa; 27.41 kDa; 34.02 kDa; 38.31 kDa;and 54.94kDa were detected.

  15. Microbial Transformation of Quercetin by Bacillus cereus

    OpenAIRE

    Rao, Koppaka V.; Weisner, Nghe T.

    1981-01-01

    Biotransformation of quercetin was examined with a number of bacterial cultures. In the presence of a bacterial culture (Bacillus cereus), quercetin was transformed into two crystalline products, identified as protocatechuic acid and quercetin-3-glucoside (isoquercitrin).

  16. Efficient activation of human T cells of both CD4 and CD8 subsets by urease-deficient recombinant Mycobacterium bovis BCG that produced a heat shock protein 70-M. tuberculosis-derived major membrane protein II fusion protein.

    Science.gov (United States)

    Mukai, Tetsu; Tsukamoto, Yumiko; Maeda, Yumi; Tamura, Toshiki; Makino, Masahiko

    2014-01-01

    For the purpose of obtaining Mycobacterium bovis bacillus Calmette-Guérin (BCG) capable of activating human naive T cells, urease-deficient BCG expressing a fusion protein composed of Mycobacterium tuberculosis-derived major membrane protein II (MMP-II) and heat shock protein 70 (HSP70) of BCG (BCG-DHTM) was produced. BCG-DHTM secreted the HSP70-MMP-II fusion protein and effectively activated human monocyte-derived dendritic cells (DCs) by inducing phenotypic changes and enhanced cytokine production. BCG-DHTM-infected DCs activated naive T cells of both CD4 and naive CD8 subsets, in an antigen (Ag)-dependent manner. The T cell activation induced by BCG-DHTM was inhibited by the pretreatment of DCs with chloroquine. The naive CD8(+) T cell activation was mediated by the transporter associated with antigen presentation (TAP) and the proteosome-dependent cytosolic cross-priming pathway. Memory CD8(+) T cells and perforin-producing effector CD8(+) T cells were efficiently produced from the naive T cell population by BCG-DHTM stimulation. Single primary infection with BCG-DHTM in C57BL/6 mice efficiently produced T cells responsive to in vitro secondary stimulation with HSP70, MMP-II, and M. tuberculosis-derived cytosolic protein and inhibited the multiplication of subsequently aerosol-challenged M. tuberculosis more efficiently than did vector control BCG. These results indicate that the introduction of MMP-II and HSP70 into urease-deficient BCG may be useful for improving BCG for control of tuberculosis.

  17. Independent transcription of glutamine synthetase (glnA2) and glutamine synthetase adenylyltransferase (glnE) in Mycobacterium bovis and Mycobacterium tuberculosis.

    Science.gov (United States)

    Hotter, Grant S; Mouat, Pania; Collins, Desmond M

    2008-09-01

    Mycobacterium bovis and Mycobacterium tuberculosis possess four glutamine synthetase homologues, two of which, glnA1 and glnA2, are required for virulence and are located on the bacterial chromosome on either side of glutamine synthetase adenylyltransferase (glnE). While glnA1 is encoded on the complementary strand, glnA2 is located 48bp upstream from glnE, raising the possibility that glnA2 and glnE may be co-transcribed. However, previous studies in M. bovis and M. tuberculosis have painted a contradictory picture of the (co)transcriptional status of glnA2 and glnE. Given the importance of the genes at the glnA1-glnE-glnA2 locus, we sought to clarify the transcriptional status of glnA2 and glnE in both M. bovis and M. tuberculosis. Reverse transcription-PCR demonstrated that glnA2 and glnE were independently transcribed in all six M. bovis and M. tuberculosis strains examined. Northern analysis of the glnA2 transcript in M. bovis AF2122/97 and M. tuberculosis H37Rv showed that it was monocistronic. These results predicted the presence of a glnE transcriptional start site in the glnA2-glnE intergenic region. An identical start site was confirmed in M. bovis AF2122/97 and M. tuberculosis H37Rv using 5' rapid amplification of cDNA ends. Typical mycobacterial -10 and -35 sequences are associated with this start site.

  18. Nested PCR detection and phylogenetic analysis of Babesia bovis and Babesia bigemina in cattle from Peri-urban localities in Gauteng Province, South Africa.

    Science.gov (United States)

    Mtshali, Phillip Senzo; Tsotetsi, Ana Mbokeleng; Thekisoe, Matlhahane Molifi Oriel; Mtshali, Moses Sibusiso

    2014-01-01

    Babesia bovis and Babesia bigemina are tick-borne hemoparasites causing babesiosis in cattle worldwide. This study was aimed at providing information about the occurrence and geographical distribution of B. bovis and B. bigemina species in cattle from Gauteng province, South Africa. A total of 268 blood samples collected from apparently healthy animals in 14 different peri-urban localities were tested using previously established nested PCR assays for the detection of B. bovis and B. bigemina species-specific genes encoding rhoptry-associated protein 1 (RAP-1) and SpeI-AvaI restriction fragment, respectively. Nested PCR assays revealed that the overall prevalence was 35.5% (95% confidence interval [CI]=± 5.73) and 76.1% (95% CI=± 5.11) for B. bovis and B. bigemina, respectively. PCR results were corroborated by sequencing amplicons of randomly selected samples. The neighbor-joining trees were constructed to study the phylogenetic relationship between B. bovis and B. bigemina sequences of randomly selected isolates. Analysis of phylogram inferred with B. bovis RAP-1 sequences indicated a close relationship between our isolates and GenBank strains. On the other hand, a tree constructed with B. bigemina gp45 sequences revealed a high degree of polymorphism among the B. bigemina isolates investigated in this study. Taken together, the results presented in this work indicate the high incidence of Babesia parasites in cattle from previously uncharacterised peri-urban areas of the Gauteng province. These findings suggest that effective preventative and control measures are essential to curtail the spread of Babesia infections among cattle populations in Gauteng.

  19. Comparative Genomics of Field Isolates of Mycobacterium bovis and M. caprae Provides Evidence for Possible Correlates with Bacterial Viability and Virulence

    Science.gov (United States)

    de la Fuente, José; Díez-Delgado, Iratxe; Contreras, Marinela; Vicente, Joaquín; Cabezas-Cruz, Alejandro; Tobes, Raquel; Manrique, Marina; López, Vladimir; Romero, Beatriz; Bezos, Javier; Dominguez, Lucas; Sevilla, Iker A.; Garrido, Joseba M.; Juste, Ramón; Madico, Guillermo; Jones-López, Edward; Gortazar, Christian

    2015-01-01

    Mycobacteria of the Mycobacterium tuberculosis complex (MTBC) greatly affect humans and animals worldwide. The life cycle of mycobacteria is complex and the mechanisms resulting in pathogen infection and survival in host cells are not fully understood. Recently, comparative genomics analyses have provided new insights into the evolution and adaptation of the MTBC to survive inside the host. However, most of this information has been obtained using M. tuberculosis but not other members of the MTBC such as M. bovis and M. caprae. In this study, the genome of three M. bovis (MB1, MB3, MB4) and one M. caprae (MB2) field isolates with different lesion score, prevalence and host distribution phenotypes were sequenced. Genome sequence information was used for whole-genome and protein-targeted comparative genomics analysis with the aim of finding correlates with phenotypic variation with potential implications for tuberculosis (TB) disease risk assessment and control. At the whole-genome level the results of the first comparative genomics study of field isolates of M. bovis including M. caprae showed that as previously reported for M. tuberculosis, sequential chromosomal nucleotide substitutions were the main driver of the M. bovis genome evolution. The phylogenetic analysis provided a strong support for the M. bovis/M. caprae clade, but supported M. caprae as a separate species. The comparison of the MB1 and MB4 isolates revealed differences in genome sequence, including gene families that are important for bacterial infection and transmission, thus highlighting differences with functional implications between isolates otherwise classified with the same spoligotype. Strategic protein-targeted analysis using the ESX or type VII secretion system, proteins linking stress response with lipid metabolism, host T cell epitopes of mycobacteria, antigens and peptidoglycan assembly protein identified new genetic markers and candidate vaccine antigens that warrant further study to

  20. The protective efficacy of cloned Moraxella bovis pili in monovalent and multivalent vaccine formulations against experimentally induced infectious bovine keratoconjunctivitis (IBK).

    Science.gov (United States)

    Lepper, A W; Atwell, J L; Lehrbach, P R; Schwartzkoff, C L; Egerton, J R; Tennent, J M

    1995-07-01

    Calves were vaccinated with cloned Moraxella bovis pili of serogroup C (experiment 1) or B (experiment 2) either as a monovalent formulation or as part of a multivalent preparation with pili of six other serogroups. Within 4 weeks of the second vaccine dose vaccinated calves and non-vaccinated controls were challenged via the ocular route with either virulent M. bovis strain Dal2d (serogroup C) or M. bovis strain 3WO7 (serogroup B) in experiments 1 and 2, respectively. Calves vaccinated with multivalent vaccines had significantly lower antibody titres than those vaccinated with monovalent preparations. Nevertheless, the levels of protection against infectious bovine keratoconjunctivitis (IBK) achieved with multivalent vaccines were 72% and 83% for the groups challenged with M. bovis strains of serogroups B and C, respectively. The serogroup C monovalent vaccine gave 100% protection against experimentally induced IBK and M. bovis isolates cultured from the eyes 6 days post-challenge were identified as belonging solely to serogroup C. Unexpectedly, only 25% protection was achieved against homologous strain challenge of calves that received the monovalent serogroup B vaccine. Furthermore, the majority of M. bovis isolates recovered from calves in this group belonged to serogroup C, as did half of those isolates cultured from the multivalent vaccinates. The remaining bacterial isolates from the latter group, together with all isolates from the non-vaccinated controls, belonged to serogroup B. Results are consistent with the hypothesis that derivatives of the serogroup B challenge inoculum had expressed serogroup C pilus antigen within 6 days of the challenge, possibly as a result of pilus gene inversion occurring in response to the presence of specific antibody in eye tissues and tears.