• Journal of Veterinary Science
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• v.22 no.2
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• pp.24.1-24.16
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• 2021

Background: Bovine tuberculosis (TB) is caused by Mycobacterium bovis, a well-known cause of zoonotic tuberculosis in cattle and deer, and has been investigated in many physiological and molecular studies. However, detailed genome-level studies of M. bovis have not been performed in Korea. Objectives: To survey whole genome-wide single-nucleotide polymorphism (SNP) variants in Korean M. bovis field isolates and to define M. bovis groups in Korea by comparing SNP typing with spoligotyping and variable number tandem repeat typing. Methods: A total of 46 M. bovis field isolates, isolated from laryngopharyngeal lymph nodes and lungs of Korean cattle, wild boar, and Korean water deer, were used to identify SNPs by performing whole-genome sequencing. SNP sites were confirmed via polymerase chain reaction using 87 primer pairs. Results: We identified 34 SNP sites with different frequencies across M. bovis isolates, and performed SNP typing and epidemiological analysis, which divided the 46 field isolates into 16 subtypes. Conclusions: Through SNP analysis, detailed differences in samples with identical spoligotypes could be detected. SNP analysis is, therefore, a useful epidemiological tracing tool that could enable better management of bovine TB, thus preventing further outbreaks and reducing the impact of this disease.

• Journal of Veterinary Science
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• v.23 no.3
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• pp.50.1-50.12
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• 2022

Background: There is an urgent need to find reliable and rapid bovine tuberculosis (bTB) diagnostics in response to the rising prevalence of bTB worldwide. Toll-like receptor 2 (TLR2) recognizes components of bTB and initiates antigen-presenting cells to mediate humoral immunity. Evaluating the affinity of antigens with TLR2 can form the basis of a new method for the diagnosis of bTB based on humoral immunity. Objectives: To develop a reliable and rapid strategy to improve diagnostic tools for bTB. Methods: In this study, we expressed and purified the sixteen bTB-specific recombinant proteins in Escherichia coli. The two antigenic proteins, MPT70 and MPT83, which were most valuable for serological diagnosis of bTB were screened. Molecular docking technology was used to analyze the affinity of MPT70, MPT83, dominant epitope peptide of MPT70 (M1), and dominant epitope peptide MPT83 (M2) with TLR2, combined with the detection results of enzyme-linked immunosorbent assay to evaluate the molecular docking effect. Results: The results showed that interaction surface Cα-atom root mean square deviation of proteins (M1, M2, MPT70, MPT83)-TLR2 protein are less than 2.5 A, showing a high affinity. It is verified by clinical serum samples that MPT70, MPT83, MPT70-MPT83 showed good diagnostic potential for the detection of anti-bTB IgG and M1, M2 can replace the whole protein as the detection antigen. Conclusions: Molecular docking to evaluate the affinity of bTB protein and TLR2 combined with ELISA provides new insights for the diagnosis of bTB.

• Korean Journal of Veterinary Research
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• v.51 no.2
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• pp.117-122
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• 2011

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.

• Journal of Microbiology
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• v.34 no.2
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• pp.198-205
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• 1996

Two proteases were partially characterized from culture filtrate of Mycobacterium, tuberculosis KIT110. Their molecular weights were approximately 200 and 180 kDa, respectively and they exhibited similar enzymatic characteristics. These enzymes were inhibited significantly by EDTA and to some extent by EGTA. Their activity was enhanced by $Ca^{2+}$ and $Mg^{2+}$ to some degree. However, $Cu^{2+}$ and $Ag^{2+}$ completely inhibited the enzyme activity at the concentration of 2.5 and 5 mM, respectively. The optimal pH was 7.0 and optimal temperature was around $40^{\circ}C$. These enzymes were rapidly inactivated at $80^{\circ}C$. Therefore, they were heat-labile, neutral metalloproteases. These enzymes exhibited antigenicity shown by their reacting with sera from the partients with pulmonary tuberculosis. These enzymes were able to degrade serum proteins including hemoglobin, bovine serum albumin, lysozyme and immunoglobulin G and structural matrix protein such as type I collagen. Therefore, these enzymes may be thought to contribute to tissue necrosis and pathogenesis during infection.

• Korean Journal of Veterinary Service
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• v.41 no.4
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• pp.239-244
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• 2018

Mycobacterium tuberculosis (M. tuberculosis) complex is the causative agent of tuberculosis (TB) in humans and bovine TB in mammalian hosts and grows very slowly. Selenium is a central molecule in nitrogen metabolism and an essential ingredient for all living cells and glutamic acid. The effects of selenium on the growth of M. tuberculosis, a representative slow-growing Mycobacterium species, were investigated and measured using the BacT Alert 3D System (MB/BacT System). Sodium selenate, at a final concentration of $10{\mu}g/mL$, reduced the average time-to detection (TTD) to 197.2 hours (95% confidence interval (CI), 179.6~214.8) from 225.1 hours (95% CI, 218~232.0) in the control culture media (P<0.05). The TTD did not increase with $\text\tiny{L}$-glutamate concentrations up to $10{\mu}g/mL$, but a significant reduction in the TTD was observed in the presence of $20{\mu}g/mL$ ${\text\tiny{L}}$-glutamate in culture media (P<0.05). In conclusion, selenate and ${\text\tiny{L}}$-glutamate enhance the growth of M. tuberculosis.

• Korean Journal of Veterinary Service
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• v.30 no.3
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• pp.393-406
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• 2007

Bovine tuberculosis is an important zoonosis worldwide. Mycobacterium bovis, the causative agent of this disease in cattle, is also a pathogen for humans and several economically important animals. The cases of tuberculosis are reported in two cow found at slaughter house located in Gwangju city. Histopathologically, in the lymph nodes, granulomas consisted of large areas of necrosis surrounded by variable thick bands of cellular infiltrate containing macrophages, Langhans-type multinucleated giant cells and lymphocytes. Lesions in the lung followed the same developmental pattern as did lesions in the lymph nodes with some exceptions. With the acid-fast staining, numerous mycobacteria were revealed in the lung and lymph nodes. M bovis was confirmed as a causative agent in these cattle using bacterial isolation and PCR and restriction fragment length polymorphism method based on a unique 12.7 kb fragment insertion sequence from the Mycobacterium tuberculosis genome and the pncA polymorphism, The insertion element IS6110 and IS1081 were present M bovis isolated from lungs and lymph nodes of cattle using PCR assay. These cases are interesting and important in public health aspect that M bovis-infected cattle were found during a routine post-mortem inspection at slaughter house.

• Korean Journal of Veterinary Service
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• v.33 no.3
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• pp.249-254
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• 2010

Deer can be one of the susceptible animals to bovine infectious diseases, and thus, may play a role either as a reservoir or amplifier host for spreading the diseases to other species such as cattle and goat. This study was conducted to determine the serum antibodies to bacterial infectious diseases for brucellosis, tuberculosis (TB), paratuberculosis (Johne's disease) in deer. Serum samples were randomly collected from 78 deer from 31 farms at Jeonbuk province, and 7 wild water deer from Jeonbuk wild animal treatment center during 2005 to 2007, respectively. Four farm deer (5.1%) showed antibodies to tuberculosis using Antigen Rapid Bovine TB Ab Test Kit. One elk (1.3%) and one wild water deer had antibodies for paratuberculosis. Antibody against Brucellosis was not detected in tube agglutination test (TAT) and enzyme-linked immunosorbent assay (ELISA). These data suggest that caution should be applied to inspection of velvet, deer blood and meat for human consumption from deer because of zoonotic bacterial diseases in deer. In addition, farmed deer can be a transmissible host for zoonotic disease to diary or raising farm.

• Korean Journal of Veterinary Service
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• v.46 no.1
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• pp.67-74
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• 2023

In the present study, the incidence of bovine tubeculosis (bTB) and brucellosis in Hanwoo (Korean native cattle) and dairy cow in Gyeongsangnam-do was investigated for three years from 2020 to 2022. The incidence bTB tested by tuberculin skin test with purified protein derivative (PPD) and gamma interferon (γ-INF) test with a commercial enzyme-linked immunosorbent assay. From 2020 to 2022, the incidence of bTB showed a decreasing trend in Hanwoo, while an increasing trend in dairy cow. In the case of Brucellosis, the positive rates for Hanwooe gradually increased. However, no brucellosis was found in dairy cow from 2020 to 2022. The increase in the incidence of these diseases is presumed to be related the small scale and poor sanitation facilities of livestock farms in Gyeongsangnam-do, and easy access of wild animals. Therefore, in order to suppress the incidence of the diseases, it is necessary to the farm scale from small to large and to strengthen sanitary facilities on farms.

• Korean Journal of Veterinary Research
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• v.41 no.4
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• pp.535-542
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• 2001

A multiplex PCR technique was developed for detecting specifically each Mycobacterium bovis, M. tuberculosis, M. avium and M. avium subsp, paratuberculosis, respectively, using clinical samples of field cattle. To apply this novel technique to clinical specimens, blood sample was obtained from live cows comprising 11 intradermal tuberculin test (ITT)-positive and 17 ITT-negative and tested by multiplex PCR. Positive results were obtained from 15 cows by the multiplex PCR, showing that 4 (23.5%) of the 17 ITT-negative cows were multiplex PCR positive. The multiplex PCR results also showed that among the 15 positive cows, 7 (46.7%) were infected with M. bovis, 1 (6.7%) with M. tuberculosis and 7 (46.7%) with M. avium. The sensitivity and specificity of multiplex PCR in comparison with those of ITT were 100% and 76.5%. The correlation between the multiplex PCR and ITT assays with blood samples was considered excellent, 85.7% agreement and ${\kappa}=0.72$. The results obtained, using reference mycobacterial strains and typed clinical samples, show that the multiplex PCR method may be a rapid, sensitive, and specific tool for the differential identification of various mycobacterial strains in a single-step assay. Therefore, multiplex PCR assay is a useful tool for early diagnosis of tuberculosis in live cattle and to identify the species or complex of mycobacterium from clinical samples.

• Korean Journal of Veterinary Service
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• v.46 no.4
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• pp.293-302
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• 2023

This study provides a comprehensive analysis of the epidemiological trends and challenges in managing tuberculosis (TB) in livestock in Korea from 2013 to 2022. Tuberculosis, caused by the Mycobacterium tuberculosis complex, is a significant zoonotic disease affecting cattle, deer, and other domesticated animals. Despite the initiation of a test-and-slaughter eradication policy in 1964, TB has continued to persist in Korean livestock, particularly in cattle and deer. This study used data from the Korea Animal Health Integrated System and provincial animal health laboratories to analyze TB incidence in various livestock including different cattle breeds and deer species. The results from 2013 to 2022 showed a peak in TB cases in 2019 with a subsequent decline by 2022. The study highlighted a significant incidence of TB in Korean native cattle and the need for amore inclusive approach towards TB testing and control in different cattle breeds. Additionally, the study underscored the importance of addressing TB in other animals such as goats, wildlife, and companion animals for a holistic approach to TB eradication in Korea. The findings suggest that while the test-and-slaughter strategy has been historically effective, there is a need for adaptation to the current challenges, and learning from successful eradiation stories on other countries like Australia. A collaborative effort involving an expanded surveillance system, active private sector participation, and robust government support essential for the efficient eradication of TB in livestock in Korea.