• Journal of Microbiology and Biotechnology
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• v.25 no.2
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• pp.255-267
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• 2015

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne's disease, a chronic debilitating disease affecting ruminants worldwide. In the present study, we aimed to determine the major gene networks and pathways underlying the immune response to MAP infection using whole-blood cells, as well as provide the potential transcriptional markers for identifying the status of MAP infection. We analyzed the transcriptional profiles of whole-blood cells of cattle identified and grouped according to the presence of MAP-specific antibodies and the MAP shed by them. The grouping was based on the results obtained by ELISA and PCR analyses as follows: i) Test1 group: MAP-negative results obtained by ELISA and positive results obtained by PCR; ii) Test2 group: MAP-positive results obtained by ELISA and negative results obtained by PCR; iii) Test3 group: MAP-positive results obtained by ELISA and positive results obtained by PCR; iv) uninfected control: MAP-negative results obtained both by ELISA and PCR analysis. The results showed down-regulated production and metabolism of reactive oxygen species in the Test1 group, activation of pathways related to the host-defense response against MAP (LXR/RXR activation and complement system) in the Test2 and Test3 groups, and anti-inflammatory response (activation of IL-10 signaling pathway) only in the Test3 group. Our data indicate a balanced response that serves the immune-limiting mechanism while the host-defense responses are progressing.

• Korean Journal of Veterinary Service
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• v.32 no.3
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• pp.251-255
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• 2009

TThis survey was carried out to investigate the prevalence of the antibody for bovine paratuberculosis (Johne's disease) in dairy cattle in Seosan-Taean area. From February to August in 2009, 254 M.RT. samples were collected from 57 farms in the regions and enzyme immuno-sorbent assay (ELISA) was conducted. Among 254 samples, 13 (5.1%) M.R.T. samples of 3 (5.2%) farms were positive by ELISA. In regional analysis, 1 (3.1%) of 34 farms in Seasan and 2 (8.6%) of 23 farms in Taean were positive in ELISA. According to the raising scale of dairy farms, the farm with below 30 heads showed the higher positive rate (2 out of 3 positive farms) than the farms with over 30 heads (1 out of 3 positive farms).

• Korean Journal of Veterinary Research
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• v.55 no.2
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• pp.89-95
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• 2015

Johne's disease, caused by Mycobacterium avium subsp. paratuberculosis (MAP), is one of the most widespread and economically important diseases in cattle. Current diagnostic methods are based on the detection of anti-MAP antibodies in serum or isolation of the causative agent. However, these techniques are often not applicable for cases of subclinical infection due to relatively low sensitivity. Therefore, finding new antigen candidates that strongly react with the host immune system had been attempted. To effectively detect infection during the subclinical stage, several antigen candidates were selected based on previous researches. Characteristics of the selected antigen candidates were analyzed using bioinformatics-based prediction tools. A total of nine antigens were selected (MAP0862, MAP3817c, MAP2077c, MAP0860c, MAP3954, MAP3155c, MAP1204, MAP1087, and MAP2963c) that have MAP-specific and/or high immune responses to infected animals. Using a transmembrane prediction tool, five of the nine antigen candidates were predicted to be membrane protein (MAP3817c, MAP3954, MAP3155c, MAP1087, and MAP1204). Some of the predicted protein structures identified using the I-TASSER server shared similarities with known proteins found in the Protein Data Bank database (MAP0862, MAP1204, and MAP2077c). In future studies, the characteristics and diagnostic efficiency of the selected antigen candidates will be evaluated.

• Korean Journal of Veterinary Service
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• v.38 no.4
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• pp.221-225
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• 2015

The bacterium causing Johne's disease (JD), Mycobacterium avium subspecies paratuberculosis (MAP), is distributed worldwide and causes progressive intestinal disease, also known as paratuberculosis, in domestic and exotic ruminants, including cattle. The subclinical form of this disease results in progressive weight loss, reduced milk production, lower slaughter value, and premature culling, with possible impacts on fertility and udder health. Eventually, infection can lead to the clinical form that manifests chronic diarrhea, emaciation, debilitation, and eventual death. JD is a major cause of economic loss in cattle industry. In this study, serological testing was conducted by ELISA to determine the prevalence of MAP in Korean native cattle of Jeju during the duration of 2010 to 2013. A total of 5,150 blood sera were collected in 111 farms and the samples were classified into four different age groups for MAP; group 1 (2 years or less), group 2 (2~4 years), group 3 (5~7 years), and group 4 (over 8 years). Overall seroprevalence of MAP was 5.2% (269/5,150), and annually from 2010 to 2013, they were 11.6% (106/915), 2.1% (10/474), 5.5% (89/1,627), and 3.0% (64/2,134), respectively. Also, the prevalence rates was determined by age groups: 48 of group 1 (17.8%), 154 of group 2 (57.3%), 48 of group 3 (17.8%), and 19 of group 4 (7.1%) were seropositive for MAP. Although seropositive samples were found in all age groups, the highest seroprevalence was found in group 2. This study showed that during the investigation the seroprevalence of MAP tended to decrease.

• Korean Journal of Veterinary Research
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• v.60 no.3
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• pp.117-122
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• 2020

Johne's disease (JD) caused by Mycobacterium avium subspecies paratuberculosis (MAP) is a chronic, wasting infectious disease in ruminants that causes enormous economic losses to the dairy and beef cattle industries. The most effective way to eradicate JD is to detect infected individuals as early as possible and remove them from the herd. However, it is difficult to detect infected individuals early with the currently using diagnostic methods. Two serological diagnostic kits commercially used worldwide and a fecal detection test were compared using 298 serum samples and feces of cattle in this study to present an efficient diagnostic method. Although there was a high correlation between the 2 serological diagnostic kits (R² = 0.7473), kit A showed a higher serological positive rate. However, the correlation between fecal tests and serological diagnosis was very low. MAP was also detected in fecal tests in many serologically negative individuals. In the periodical diagnosis of JD, MAP was detected in the feces of only cows with the higher antibody titer to MAP. These results suggest that for effective eradication of JD, early detection of infected individuals by fecal tests together with the serological tests currently in use and by removal of infected individuals are needed.

• Korean Journal of Veterinary Research
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• v.44 no.4
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• pp.669-676
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• 2004

To evaluate the diagnostic accuracy of two commercial ELISA tests (Allied- and CSL-ELISA) for the diagnosis of Mycobacterium paratuberculosis in cattle, Meta-analysis using English language papers published during 1990-2001 was performed. Diagnostic odds ratios (DOR) were analyzed using regression analysis together with summary receiver operating characteristic (ROC) curves. The difference in diagnostic performance between the two ELISA systems was evaluated by using linear regression. Publication bias was assessed by funnel plot and linear regression. The pooled sensitivity and specificity were 44% (95% CI, 38 to 51) and 98% (95% CI, 96 to 99) for the random-effect model. The DOR between studies was heterogeneous. The area under the fitted ROC curve (AUC) was 0.72 for the unweighted and 0.77 for the weighted model. Maximum joint sensitivity and specificity for the unweighted and weighted model from their summary ROC curve were 70% and 75%, respectively. Based on the fitted model, at a specificity of 95%, sensitivity was estimated to be 52% for the unweighted and 57% for the weighted model. From the final multivariable model study characteristic, the country was the only significant variable with an explained component variance of 13.3%. There were no significant differences in discriminatory power, sensitivity, and specificity between the two ELISA tests. The overall diagnostic accuracy of two commercial ELISA tests was moderate, as judged by the AUC, maximum joint sensitivity and specificity, and estimates from the fitted model and clinical usefulness of the tests for screening program is limited because of low sensitivity and heterogeneous of DOR. It is, therefore, recommended to use ELISA tests as a parallel testing with other diagnostic tests together to increase test sensitivity in the screening program.

• Journal of Life Science
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• v.22 no.6
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• pp.794-798
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• 2012

Paratuberculosis (Johne's disease), a chronic wasting disease caused by Mycobacterium avium subspecies paratuberculosis (MAP), is a major cause of economic loss in the cattle industry. In Korea, national monitoring of breeding stock for MAP has been implemented. In this study, we report the results of serological testing to determine the prevalence of MAP in breeding stock of Korean native and dairy cattle during 2008 and 2009. A total of 3,927 serum samples were submitted (3,692 Korean native cattle and 235 dairy cattle) to Animal Disease Diagnostic Division, Animal, Plant and Fisheries Quarantine and Inspection Agency. The samples were classified into four different age groups for MAP; group 1 (${\leq}2$ year, n = 1,509), group 2 (>2 years to ${\leq}3$ years, n = 486), group 3 (>3 years to ${\leq}4$ years, n = 441), and group 4 (>4 years, n = 1,491). Overall seroprevalence of MAP in this study was 0.5% (21/3,927), which was much lower than that of conventional cattle (1.2-16.4%) in Korea. Also, the seroprevalence was determined by age groups: three of group 1 (0.2%), two of group 2 (0.4%), three of group 3 (0.7%), and 13 of group 4 (0.9%) were seropositive for MAP, respectively. Although seropositive samples were found in all age groups, the seroprevalence tended to increase with age. Our study showed that the seroprevalence of MAP in pure-bred breeding dairy cattle (0%) was lower than that in pure-bred breeding Korean native cattle (0.6%).

• BMB Reports
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• v.47 no.2
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• pp.115-120
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• 2014

In this study, we show that Mycobacterium avium subsp. paratuberculosis MAP1305 induces the maturation of bone marrow-derived dendritic cells (BMDCs), a representative antigen presenting cell (APC). MAP1305 protein induces DC maturation and the production of pro-inflammatory cytokines (Interleukin (IL)-6), tumor necrosis factor (TNF)-${\alpha}$, and IL-$1{\beta}$) through Toll like receptor-4 (TLR-4) signaling by directly binding with TLR4. MAP1305 activates the phosphorylation of MAPKs, such as ERK, p38MAPK, and JNK, which is essential for DC maturation. Furthermore, MAP1305-treated DCs transform naive T cells to polarized $CD4^+$ and $CD8^+$ T cells, thus indicating a key role for this protein in the Th1 polarization of the resulting immune response. Taken together, M. avium subsp. paratuberculosis MAP1305 is important for the regulation of innate immune response through DC-mediated proliferation of $CD4^+$ and $CD8^+$ T cells.

• Journal of Veterinary Clinics
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• v.35 no.1
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• pp.7-9
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• 2018

Mycobacterium avium subspecies paratuberculosis (MAP) causes a significant economic burden in the animal production industry. The purpose of this study was to investigate the prevalence of MAP in the feces of wild duck populations residing along a riverside close to farms in the center of Korea. From wild Spot-billed (Anas poecilorhyncha) and Mallard (Anas platyrhynchos) ducks, 128 fecal samples were collected and analyzed using multiplex real-time PCR, sequencing, and nested PCR to confirm the presence of the organism. The molecular analyses showed that 44 samples (34.4%) were positive for MAP, suggesting a high prevalence of MAP in the wild duck population. Considering the nature and habitat of wild ducks, this result suggests that the organism was introduced from contaminated water from waste of nearby farms, and that the wild ducks may act as a transmitter of the organism to other wild birds or livestock.

• Biomedical Science Letters
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• v.6 no.1
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• pp.65-72
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• 2000

In order to improve the early diagnosis of Johne's disease in ruminants, duplex polymerase chain reaction system for the detection of the etiologic agent of M. paratuberculosis and for the differentiation of other mycobacterial animal pathogens, such as M. bovis and M. avium, was applied. Genomic DNAs were purified from peripheral blood monocytes or milk macrophages and were used as templates in the duplex PCR. Detection of Mycobacterium spp. in the specimen was carried out by PCR using primer set specific to the mycobacterial 16S rDNA. And then, mycobacterial DNA-positive specimens were further differentiated with duplex PCR system which was composed of primer sets specific to 16S rDNA of M. avium complex and Is900 gene of M. paratuberculosis. The results were re-confirmed by Southern blot hybridization with oligonucleotide specific to the internal sequence of IS900 PCR amplicons. The applicability of this duplex PCR system was evaluated with DNAs extracted from clinical specimens of peripheral blood monocytes and milk macrophages. In summary, the duplex PCR amplification system described in this experiment is promising molecular technique for the early diagnosis of Johne's disease in ruminants.