• Korean Journal of Veterinary Service
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• v.30 no.2
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• pp.251-258
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• 2007

Bovine brucellosis has occurred for years in Gyeonggi province under the national test and slaughter scheme. The serum agglutination test (SAT) is a diagnostic tool to confirm the disease despite the argument on its specificity. We selected 8 farms where only one or two individuals were diagnosed as brucellosis through SAT at the primary regular herd check and isolated the causative organism and characterized the species by species-specific PCR. The pathogen isolation was successful in 6 farms out of 8 farms by microbiological culture, showing the successful rate of 75%. The isolation rate of the causative organism represents 70% from supra-mammary lymph node and 60% from uterine tissues. They were characterized as Brucella abortus biovar 1 after biotyping by PCR, showing the fragment of 498 bp. Five of 8 farms were diagnosed as brucellosis two to four times more over the intervals of two or three months. Here in this study we briefly showed the correlation of the sporadic outbreak of brucellosis tested by SAT and the isolation of the causative organism. Moreover one or two reactors against brucellosis among considerable size of herd may indicate that SAT failed to detect potentially infected individuals in the incubation stage or chronic phase of the disease.

• Journal of Food Hygiene and Safety
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• v.29 no.3
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• pp.165-169
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• 2014

Salmonella spp. and Brucella spp. are associated with considerable diseases of both humans and animals. In addition, these microorganisms cause the economic loss in animal farming and food industry. In this study, the disinfection efficacy of a commercial disinfectant, composed to povidone-iodine was evaluated against S. typhimurium and B. ovis. A bactericidal efficacy test by broth dilution method was used to determine the lowest effective dilution of the disinfectant following exposure to test bacteria for 30 min at $4^{\circ}C$. The disinfectant and test bacteria were diluted with hard water (HW) or organic matter suspension (OM) according to treatment condition. On HW condition, the bactericidal activity of the disinfectant against S. typhimurium and B. ovis was 400 and 150 fold dilutions, respectively. On OM condition, the bactericidal activity of the disinfectant was 5 and 20 fold dilutions against S. typhimurium and B. ovis, respectively. As the disinfectant composed to povidine-iodine possesses bactericidal efficacy against animal pathogenic bacteria such as S. typhimurium and B. ovis, the disinfectant solution can be used to control the spread of bacterial diseases.

• Korean Journal of Veterinary Research
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• v.38 no.2
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• pp.345-352
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• 1998

The diagnosis of brucellosis is currently based on serological and microbiological tests. However, the microbiological isolation and identification have several disadvantages such as time-consuming and laborious, and the serological methods have been reported to cross-react with antigens other than those from Brucella spp. To develop a sensitive and rapid diagnostic method for detection of Brucella species, the genus-specific primers were designed and synthesized from the sequence of gene encoding a 31kDa cell surface protein(BCSP) and a 36kDa outer membrane protein(OMPB) of B abortus. The amplified 711bp and 982bp DNA fragments were only visible in each species of Brucella by PCR method using the BCSP and OMPB primers, respectively. However, PCR product was not obtained with DNA from other Gram-negative bacteria. As little as 1pg of the B abortus genomic DNA could be detected by this PCR method. Using the PCR technique, semen samples from 185 bulls of Brucella-seronegative herds in Cheju island were examined for comparison of this PCR method with conventional methods in 1995. The semen samples from 5 bulls were positive by culture method and PCR, and one was positive and 5 were suspect by semen plasma agglutination test. However, the semen samples obtained from 177 bulls were negative by semen plasma agglutination, culture and PCR methods in 1996. The results of comparison tests suggested that PCR was a better test than agglutination test against semen of bulls. This study indicated that the PCR technique was a valuable for the diagnosis of bovine brucellosis, particulary in bull semens.

• Journal of Food Hygiene and Safety
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• v.31 no.4
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• pp.299-303
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• 2016

Salmonella spp. and Brucella spp. can cause considerable diseases on both humans and animals. In addition, these microorganisms cause the economic loss in animal farming and food industry. In this study, the disinfection efficacy of a disinfectant spray, composed to grapefruit seed extract, citric acid, malic acid and benzalkonium chloride, was evaluated against S. Typhimurium and B. ovis. A bactericidal efficacy test by broth dilution method was used to determine the lowest effective dilution of the disinfectant following exposure to test bacteria for 30 min at $4^{\circ}C$. The disinfectant and test bacteria were diluted with hard water (HW) or organic matter suspension (OM) according to treatment condition. On HW condition, the bactericidal activity of the disinfectant spray against S. Typhimurium and B. ovis was 5 and 4 fold dilutions, respectively. On OM condition, the bactericidal activity of the disinfectant spray was 2 and 1 fold dilutions against S. Typhimurium and B. ovis, respectively. As the disinfectant spray possesses bactericidal efficacy against foodborne pathogens such as S. Typhimurium and B. ovis, the disinfectant spray can be used to control the spread of bacterial diseases.

• Proceedings of the Microbiological Society of Korea Conference
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• 2008.05a
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• pp.144-146
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• 2008

• Korean Journal of Veterinary Service
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• v.33 no.2
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• pp.121-128
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• 2010

Brucellosis is a serious zoonosis, recognized worldwide. It primarily affects animals, which act as reservoirs for human infection as well as being of economic significance to the agri-food industry. Bangladesh has been reported as an endemic area for brucellosis. So a cross sectional study was conducted to determine the seroprevalence and potential risk factors of brucellosis in cattle in Dinajpur and Mymensingh districts of Bangladesh. A total of 182 cattle were examined by Rose Bengal Plate Test (RBPT) between September 2008 and October 2009. Then Positive, doubtful, and negative samples were further confirmed with slow agglutination test (SAT) and both indirect and competitive enzyme linked immunosorbent assay (iELISA and cELISA). A questionnaire was used to collect epidemiological information of the animals. The overall animal-level prevalence was 3.30%. Brucellosis seroprevalence was higher (4.76% by cELISA) in cattle above 48 months than those under 48 months. Female showed higher seroprevalence (10.67%) than male (6.25%). Higher seroprevalence was also found in cattle bred naturally (20.0%) than artificially (8.77%) and cattle that aborted or with previous abortion record (22.22%) showed higher seroprevalence than non-aborted (7.69%). The sensitivity of RBT and SAT was found 100% as compared to cELISA standard test, whereas specificity of RBT (95.35%) was higher than that of SAT (94.32%).

• Korean Journal of Veterinary Research
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• v.38 no.2
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• pp.328-335
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• 1998

Outer membrane proteins(OMPs) of Brucella abortus 1119-3 strain were extracted by Triton X-100 treatment, and fractionated by DEAE-cellulose column chromatography and Sephacryl S-300 column chromatography. The antigenic proteins in these fractions were identified by Western blot analysis. In Western blot analysis, a single band(38kDa) was observed in the DEAE fractions from 36th fraction to 38th fraction against sera of cattle infected with B abortus. And other fractions have several bands. However, the Sephacryl S-300 fractions exhibited a total of 3 peaks of proteins with a broad range from about 30 to 116kDa. In order to characterize further, the extracted OMPs and the DEAE fractions were analyzed by two dimensional polyacrylamide gel electrophoresis(2-DE) and Western blot using serum from naturally infected cattle with Brucella spp. The 2-DE immunoblots of DEAE fraction showed immunoreactive spots more than twenty two. The major protein spots have ranging from about 32 to 47kDa. The pI values of the spots were detected from pH 4.7 to 5.4. Among the major protein spots, the 38kDa protein which is a specific antigen, located at the point of approximately a pI 4.8.

• Korean Journal of Veterinary Service
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• v.33 no.2
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• pp.113-119
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• 2010

Brucellosis is a major zoonosis caused by Gram negative facultative intracellular bacterial organisms of the genus Brucella that are pathogenic for a wide variety of animals and human beings. Because of its economic impact on animal health and the risk to the human population,most countries have a brucellosis control program. Brucellosis is also an economically important andprevalent disease in Bangladesh. The accurate and prompt diagnosis is very important in controlling and eradicating of the disease in animals. The present study was undertaken to determine the seroprevalence of brucellosis in cattle in Mymensingh and Patuakhali district of Bangladesh. A total of 120 serum samples were collected from the two districts along with a questionnaire related to the epidemiology of the disease. The sampleswere screened by using slow agglutination test and conformed by indirect enzyme linked immunosorbent assay. The overall seroprevalence of brucellosis in cattle was 5% and it was observed that, a higher prevalence of Brucella was found in female than male, through natural breeding than artificial insemination (AI) and animal above 4 years old are highly susceptible than younger ones. Higher prevalence was found in aborted animals in comparison with non aborted animal. Finally, the study revealed that the female animal has more susceptible to brucellosis and healthy semen should be used for AI.

• Korean Journal of Veterinary Research
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• v.51 no.1
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• pp.15-20
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• 2011

In this study, we isolated 12 of Brucella (B.) spp. from cattle, which have been positive in Rose Bangal test and tube agglutination test in Gyeongbuk province in 2009. According to AMOS PCR analysis, isolated 12 strains were identified as B. abortus. Murine derived macrophage, RAW 264.7 cells, were infected with isolated 12 strains or reference strain (B. abortus 544), and bacterial internalization were characterized. According to these results, we divided the isolated strains into the following three groups: class I, lower internalization than that of B. abortus 544; class II, similar internalization to that of that of B. abortus 544; class III, higher internalization than that of B. abortus 544 within RAW 264.7 cells. Furthermore, intracellular growth, bacterial adherent assay, LAMP-1 colocalization, virulence in mice and surface protein pattern were characterized. From these results, representative strains of class III showed lower LAMP-1 colocalization, higher adherent efficiency, higher virulence in mice than those of B. abortus 544, and showed different pattern of surface proteins. These results suggest that B. abortus field strains, isolated from cattle in Korea, possess various virulence properties and higher internalization ability of field strain may have an important role for its virulence expression.

• Korean Journal of Veterinary Service
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• v.33 no.2
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• pp.135-141
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• 2010

Bovine brucellosis, an important zoonosis, is diagnosed with serological tests such as the RBT, TAT using inactivated whole bacterial cells or bacterial lipopolysaccharide (LPS) antigen in Korea. However, a strong cross-reaction between Brucella spp. and Yersinia enterocolitica O9 in these tests has seriously complicated the diagnosis of animal brucellosis because Brucella spp. shares common antigenic determinants with Y. enterocolitica O9 in the smooth LPS region. In this study, Brucella-field strains were isolated from Brucellapositive Hanwoo in Kimhae, Korea and outer membrane protein (omp) which has low cross-reaction with Y. enterocolitica O9 and high immunogenicity was extracted from the field strains Then we compared ELISA using the extract with RBT-TAT. Fifteen field strains were isolated from 47 supra-mammary-lymph nodes, which were collected from 18 farms. Isolation rate was 32%. Brucella-specific antigen was identified by performing SDS-PAGE or Western blotting on extracted omp with at 0.5% n-lauroylsarcosine One hundred and ninety-two serum-samples were used in the experiment: 142 negative and 50 positive samples verified by RBT-TAT. According to ELISA results, 127 samples were negative and 15 appeared positive among 142 negatives by RBT-TAT, while 42 samples were positive and 8 were negative among 50 positives by RBT-TAT. Therefore, it showed 89.4% of specificity and 84% of sensi-tivity. Through the current experiments, we could set up an ELISA based on the omp which has low cross-reaction and high immunogenicity and concluded that the omp could be a good material for accurate diagnosis of bovine brucellosis.