Bovine tuberculosis, which is caused by Mycobacterium bovis, a member of the M. tuberculosis complex, is a zoonosis transmitted through the inhalation of infected droplets or the ingestion of raw milk. Human bovine tuberculosis has been reported rarely in most countries since the introduction of pasteurized milk and M. bovis eradication programs. However, it has been reported in other areas with poorly controlled programs. We encountered a case of localized empyema due to M. bovis infection in a pregnant female immigrant from Vietnam. We report this case with a brief review of the related literature.
Eating ; Emigrants and Immigrants ; Empyema ; Female ; Humans ; Inhalation ; Milk ; Mycobacterium ; Mycobacterium bovis ; Tuberculosis ; Tuberculosis, Bovine ; Vietnam

Bovine tuberculosis, which is caused by Mycobacterium bovis, a member of the M. tuberculosis complex, is a zoonosis transmitted through the inhalation of infected droplets or the ingestion of raw milk. Human bovine tuberculosis has been reported rarely in most countries since the introduction of pasteurized milk and M. bovis eradication programs. However, it has been reported in other areas with poorly controlled programs. We encountered a case of localized empyema due to M. bovis infection in a pregnant female immigrant from Vietnam. We report this case with a brief review of the related literature.
Eating ; Emigrants and Immigrants ; Empyema ; Female ; Humans ; Inhalation ; Milk ; Mycobacterium ; Mycobacterium bovis ; Tuberculosis ; Tuberculosis, Bovine ; Vietnam

The prevalence of mastitis, milk quality and health risks associated with milk consumption were investigated on 96 randomly selected traditional herds in Dodoma rural and Mvomero districts of Tanzania. Mastitis was investigated based on clinical signs, microbiology and California mastitis test (CMT), while milk quality was evaluated using total viable count (TVC)and total coliform count (TCC). Animals were tested for tuberculosis using a single comparative intradermal tuberculin test. The prevalence of subclinical mastitis based on CMT was low (8.3%). The major isolates were Staphylococcus aureus (35.3%), other staphylococci (20.8%), coliforms (27.7%), microcci (5.8%) and streptococci (9.8%). The average TVC of milk in Dodoma rural district (1.0 x10(7)+/-3.4 x10(7))was significantly higher than that in Mvomero district (8.9x10(5) 3.5x10(6)) (p<0.001)and the proportion of TCC-positive samples in Dodoma (70.7%)were significantly higher (p<0.001) than that of Mvomero sample(20.8%). Whereas no tuberculin reactor animal was detected in the study animals, atypical mycobacteria were isolated from milk and one sample from Dodoma had Mycobacterium tuberculosis. Knowledge on health risks associated with milk consumption was low (20.8%). It is concluded that lack of awareness on health risks associated with milk consumption amongst rural communities needs to be addressed in order to safeguard their health.
Animals ; Cattle ; Female ; Humans ; Mastitis, Bovine/*epidemiology ; Milk/*microbiology/*standards ; Prevalence ; Public Health ; Tanzania/epidemiology ; Tuberculosis, Bovine/*epidemiology

Bovine tuberculosis caused by Mycobacterium bovis is a major economic problem in several countries. Antibody responses are useful indicators of M. bovis infection of cattle. To overcome drawback of serological tests with low sensitivity, identification and characterization of multiple serodiagnostic antigens has been required. In this study, the antigens with strong antibody reactivity were searched using fractionation of M. bovis culture filtrate proteins and probing with sera from M. bovis-infected cattle. Twelve proteins which have not previously been described as serologic targets were identified and six proteins among them were expressed in Escherichia coli. The mycobacterial lipoarabinomannan (LAM) with strong seroreactivity in cattle was identified and purified. IgG and IgA responses against the newly identified proteins, the seroreactive proteins with strong antibody reactivity in human tuberculosis, and LAM were compared in M. bovis-infected and non-infected cattle as well as in field samples. In general, sensitivity of the tested antigens was higher in M. bovis-infected cattle than purified protein derivative (PPD) (+) field samples. Although a diverse reactivity and sensitivity according to the antigens were shown, the diagnostic utility of both IgA and IgG antibody to the antigens was similar in M. bovis-infected cattle but utility of IgG antibody was superior to that of IgA in field samples. The antigen with the highest diagnostic value was LAM in both the groups. Other antigens with considerable diagnostic utility were BCG_3488c, BCG_2330, Antigen 85, HspX, and Rv3593 when considered the sensitivity and area under the receiver characteristic curve (AUC) value. These antigens may be valuable candidates to be included in a cocktail test kit for bovine tuberculosis diagnosis.
Animals ; Antibody Formation ; Cattle* ; Diagnosis ; Escherichia coli ; Humans ; Immunoglobulin A ; Immunoglobulin G ; Mycobacterium bovis ; Serologic Tests ; Tuberculosis ; Tuberculosis, Bovine*

BACKGROUND: The inhibition rates for nucleic acid tests of Mycobacterium tuberculosis have been reported to range from less than 1% to more than 10%. Specimen dilution, boiling, addition of bovine serum albumin (BSA), and a silica membrane can be used to override amplification inhibitors in nucleic acid tests of M. tuberculosis. The inhibition rate for real-time PCR of M. tuberculosis (COBAS TaqMan MTB test; Roche Diagnostics, Manheim, Germany) and effective strategies to override PCR inhibitors were investigated in this study. METHODS: The inhibition rate for COBAS TaqMan MTB test was investigated in 980 clinical specimens. The effectiveness of PCR inhibitor removal by repeated run, dilution, boiling, addition of BSA, and use of silica membrane were evaluated in the inhibited specimens. RESULTS: Inhibitory substances were present in 4.1% of specimens (40/980). Among 40 inhibited specimens, inhibitory substances were removed in 12 (30%), 30 (75%), 27 (67.5%), 25 (62.5%) and 12 (30%) specimens with repeated run, dilution, addition of RBS, boiling and use of silica membrane, respectively. CONCLUSION: The overall inhibition rate for the COBAS TaqMan MTB test was 4.1%. Dilution, boiling and addition of BSA were shown to be more effective than repeated run and use of silica membrane for removal of PCR inhibitors. A combination of two methods might be useful and should be studied in the future.
Membranes ; Mycobacterium ; Mycobacterium tuberculosis ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction ; Serum Albumin, Bovine ; Silicon Dioxide ; Tuberculosis

Bovine tuberculosis (bTB), caused primarily by Mycobacterium bovis, continues to exert an economic loss, even in countries with active control measures, and is one of zoonotic diseases enable to be transmitted to human. The control and eradication of bTB are mainly based on a test and slaughter policy and/or abattoir surveillance. Various factors including limitation of diagnostic tests have been considered as major constraints to eradication. Single intradermal test (SIT) is the official diagnostic test. New diagnostic methods are needed to be developed, because of limitations of the test. In the present study SIT was compared with single intradermal comparative cervical test (SICCT) and interferon (IFN)-gamma assay. There was very low correlation between SIT and SICCT. However, high correlation was shown between SIT and IFN-gamma assay while no correlation was observed between SICCT and IFN-gamma assay. Therefore, our results suggest the possibility of replacement of SIT with IFN-gamma assay for the diagnosis of bovine tuberculosis.
Abattoirs ; Animals ; Cattle ; Diagnostic Tests, Routine ; Humans ; Interferon-gamma ; Interferons ; Intradermal Tests ; Mycobacterium bovis ; Skin ; Skin Tests ; Tuberculin ; Tuberculosis, Bovine

We established an ELISPOT for bovine interferon-gamma (BoIFN-γ), and applied it in the diagnosis of bovine tuberculosis (bTB). Monoclonal antibodies that can bind with native BoIFN-γ were screened as the coating antibody and detecting antibody. After optimization of detecting conditions including coating antibody concentration, cell number, and detecting antibody concentration, the ELISPOT assay was established. Peripheral mononuclear cells (PBMCs) isolated from 30 cows were co-cultured with PPD, and detected with the ELISPOT assay. The optimal conditions of ELISPOT assay were 2.5 μg/mL coating antibody 2G5, 2.5 x 10(5) cells/well, and 1 μg/mL detecting antibody Bio-5E11. In these 30 cows tested both with the ELISPOT assay and the BOVIGAM kit, 11 cows were proved to be positive in ELISOPT assay with the sensitivity of 78.6%, and 12 cows were proved to be negative in ELISOPT assay with the specificity of 75%. The ELISPOT assay for BoIFN-γ could be used to detect bTB efficiently and it might be an alternative method for the diagnosis of bTB.
Animals ; Antibodies, Monoclonal ; Cattle ; Enzyme-Linked Immunospot Assay ; veterinary ; Female ; Interferon-gamma ; isolation & purification ; Sensitivity and Specificity ; Tuberculosis, Bovine ; diagnosis

Bovine tuberculosis (TB) is a major zoonosis that's caused by Mycobacterium bovis (M. bovis). Being able to detect M. bovis is important to control bovine TB. We applied a molecular technique, the variable number tandem repeat (VNTR) typing method, to identify and distinguish the M. bovis isolates from Gyeonggi-do, Korea. From 2003 to 2004, 59 M. bovis clinical strains were isolated from dairy cattle in Gyeonggi-do, Korea, and these cattle had tuberculosis-like lesions. Twenty-four published MIRUVNTR markers were applied to the M. bovis isolates and ten of them showed allelic diversity. The most discriminatory locus for the M. bovis isolates in Korea was QUB 3336 (h = 0.64). QUB 26 and MIRU 31 also showed high discriminative power (h = 0.35). The allelic diversity by the combination of all VNTR loci was 0.86. Six loci (MIRU 31, ETR-A and QUB-18, -26, -3232, -3336) displayed valuable allelic diversity. Twelve genotypes were identified from the 59 M. bovis isolates that originated from 20 cattle farms that were dispersed throughout the region of Gyenggi-do. Two genotypes [designation index (d.i.) = e, g] showed the highest prevalence (20% of the total farms). For the multiple outbreaks on three farms, two successive outbreaks were caused by the same genotype at two farms. Interestingly, the third outbreak at one farm was caused by both a new genotype and a previous genotype. In conclusion, this study suggests that MIRU-VNTR typing is useful to identify and distinguish the M. bovis isolates from Gyeonggi-do, Korea.
Animals ; Cattle ; DNA Primers/genetics ; *Genetic Variation ; Genotype ; Korea/epidemiology ; Minisatellite Repeats/*genetics ; Mycobacterium bovis/*genetics ; Prevalence ; Tuberculosis, Bovine/*epidemiology/*microbiology

Bovine tuberculosis is a chronic contagious disease responsible for major agricultural economic losses. Abattoir monitoring and trace-back systems are an appropriate method to control bovine tuberculosis, particularly in beef cattle. In the present study, a trace-back system was applied to bovine tuberculosis cases in Korean native Hanwoo beef cattle. Bovine tuberculosis was detected in three index beef cattle during abattoir monitoring in Jeonbuk Province, Korea, and the original herds were traced back from each index cow. All cattle in each original herd were subjected to tuberculin skin test. The positive rates in the tuberculin skin test were 64.6% (62 of 96), 4.8% (2 of 42), and 8.1% (3 of 37) at farms A, B, and C, respectively. On post-mortem examination of 56 tuberculin-positive cattle, 62% had granulomatous lesions, and Mycobacterium bovis was cultured from 40 (71.4%) of the cattle. Molecular typing by spoligotyping and the mycobacterial interspersed repetitive unit-variable-number tandem repeat assay revealed the genotype of the M. bovis strains from the index cattle were same as the M. bovis genotype in each original herd. The results suggest that tracing back from index cattle to the original herd is an effective method to control bovine tuberculosis in beef cattle.
Abattoirs ; Agriculture ; Animals ; Autopsy ; Cattle ; Disease Outbreaks ; Genotype ; Jeollabuk-do ; Korea ; Methods ; Molecular Typing ; Mycobacterium bovis ; Red Meat ; Skin Tests ; Tandem Repeat Sequences ; Tuberculin ; Tuberculosis, Bovine

The interferon-gamma (IFN-gamma) assay is employed as a complementary diagnostic test for bovine tuberculosis (BTB) in many countries. To simplify this assay, we established a 96-well plate format using the ESAT-6 and CFP-10 antigens and then employed it to determine the extent of Mycobacterium (M.) bovis infection in dairy herds with a history of BTB outbreaks in a country where only selective culling is practiced. The sensitivity and specificity of this IFN-gamma assay were 85.9% and 100%, respectively, based on comparison with the conventional single intradermal tuberculin test (SIDT). The IFN-gamma assay was also positive in 30.4% and 36.8% of SIDT-negative animals from herds with recent and remote BTB outbreaks, respectively. Of 14 SIDT-negative, IFN-gamma positive cattle, five (35.7%) were culture positive and an additional six were positive based on a polymerase chain reaction-based test for M. bovis. Therefore, the IFN-gamma assay has the potential to serve as a specific and sensitive test for M. bovis infection in dairy cattle. Further, the results indicated that a substantial portion of SIDT-negative animals in herds with previous BTB outbreaks were actually infected with M. bovis. Accordingly, the present selective-culling strategy may require modifications to include this more sensitive assay.
Animals ; Antigens, Bacterial/*diagnostic use ; Bacterial Proteins/diagnostic use ; Cattle ; Female ; Interferon-gamma Release Tests/*veterinary ; Mycobacterium bovis/*isolation & purification ; Polymerase Chain Reaction/veterinary ; Republic of Korea/epidemiology ; Tuberculosis, Bovine/*diagnosis/*epidemiology/microbiology