• 대한수의학회지
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• 제43권1호
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• pp.67-75
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• 2003

A total of 260 dogs were randomly selected from two different treed kennels that brucellosis has occurred (group 1, 126 dogs), and random selected breed kennel (group 2, 134 dogs), and monitored for Brucella canis (B. canis) by 2-mercaptoethanol rapid slide agglutination test (2ME-RSAT) and bacterial culture method. For the differentiation, PCR-RFLP using omp-31, wbkA and per genes used for 52 of B canis strains (strain I) isolated in this study and 3 of B. canis strains (strain II) isolated in 1994 in Korea. 2ME-RSAT revealed that 63/126 dogs (50.0%) and 12/134 dogs (9.0%) were positive in group I and group II, respectively. Bacterial culture revealed that 47/126 dogs (37.3%) and 5/134 dogs (3.7%) were positive in group I and group II, respectively. As the results of PCR-RFLP, $\underline{omp}-31$ was amplified from all Brucella spp, except B. abortus. All B. canis isolates showed unique PCR-RFLP pattern following digestion with Bmel8I. However, all Brucella spp. showed the same PCR-RFLP pattern following digestion with SalI. PCR-RFLP analysis of wbkA revealed that all Brucella spp. showed the same pattern following digestion with HindIII. PCR-RFLP analysis of per revealed that B. abortus 544 and B. melitensis 63/9 showed the same pattern, but different from B. suis and B. canis following digestion with HindIII.

• 한국동물위생학회지
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• 제33권1호
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• pp.29-35
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• 2010

This study was performed to test the hypothesis that Brucella abortus strain RB51 (SRB51) might protect Korean indigenous mongrel dog against challenge with either virulent B. abortus biotype 1 or B. canis. A total of 12 Korean mongrel dogs were divided into four groups (Group A, B, C and D). Dogs belonging to Group A and C were inoculated subcutaneously with $1{\times}10^9$ CFU of SRB51 in 1ml of sterile phosphate buffered saline (PBS). Dogs of Group B and D were inoculated subcutaneously with 1ml of sterile PBS as control. At 12 weeks post vaccination, dogs of Group A and B were challenged by oral inoculation of virulent strain of B. canis ($5.0{\times}10^9$ CFU) and dogs of Group C and D were challenged by oral inoculation of virulent strain of B. abortus biotype 1 ($4.4{\times}10^{10}$ CFU). The serum antibodies titers in all dogs were monitored at regular interval for eight weeks after challenge (AC) by standard tube agglutination test, plate agglutination test, rose bengal test, 2-mercaptoethanol rapid slide agglutination test and 2-mercaptoethanol tube agglutination test. No antibody titers in Group A and C was detected. Also, the challenge strains were not found from blood of all dogs of Group A and C from 1 week AC till the end of the experiment by culture and modified AMOS-PCR, whereas B. canis and B. abortus challenge strains were detected from blood of Group B and D, respectively. In addition, neither of two challenge bacteria was recovered from liver, spleen, kidneys, lymph nodes and reproductive tracts of Group A and C dogs after postmortem. However, B. canis and B. abortus challenge strains were isolated from these tissues of Group B and D, respectively. These data suggest that SRB51 could be a promising vaccine candidate for immunizing dogs to control canine brucellosis caused by B. canis or B. abortus.

• 한국동물위생학회지
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• 제34권2호
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• pp.159-166
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• 2011

This study was carried out to develope enzyme-linked immunosorbent assay (ELISA) for detection of canine brucellosis in dogs experimentally inoculated with Brucella abortus 1119-3 and B. canis RM666. Groups A, B and C of dogs (each group consisting of three dogs) were orally inoculated with approximately $5{\times}10^9$ colony-forming units of B. abortus and B. canis, and with sterile pyrogen-free PBS, respectively. The animals were monitored at regular intervals upto the 12th week post inoculation (PI) by standard tube agglutination test (STAT), plate agglutination test (PAT), Rose Bengal test (RBT), 2-mercaptoethanol rapid slide agglutination test (2ME-RSAT) and ELISA. The induced antibody titers in group A dogs were detected from the first week PI to the eighth week PI in STAT, PAT and RBT using the inactivated whole cells of B. abortus 1119-3 as antigens, while no sera in groups B and C dogs reacted with the antigens. In 2ME-RSAT using whole cells of B. canis M-strain as antigens, the induced antibody titers in group B dogs were observed at the second week PI and persisted for the 12th week PI, while sera of groups A and C dogs did not react with the whole cells. In ELISA using cytoplasmic fractions antigen of B. abortus 1119-3, the mean optical density of antibodies in groups A and B was detected from the first and second weeks PI, respectively, and persisted for 12th week PI, while sera of group C did not cross-react with the fractions antigen. However, in ELISA using the hot saline extracts of B. canis M- as an antigen, the induced antibody titers in only group B dogs were detected from second week PI and persisted for until the end of this study. These results indicate that the ELISA using B. abortus 1119-3 cytoplasmic fractions as antigens can be a good candidate for detection of brucellosis by B. abortus as well as B. canis in dogs.

• 한국동물위생학회지
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• 제32권4호
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• pp.335-341
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• 2009

Canine brucellosis produce abortions and infertility in dogs and is currently diagnosed by serological methods such as rapid slide agglutination test with 2-mercaptoethanol (2-ME RSAT) and immunochromatographic assay (ICA). Bacterial isolation is considered gold standard for Brucella diagnosis and the polymerase chain reaction (PCR) is an alternative method to bacterial isolation. A total of 36 whole blood samples were collected from dogs reared in area of Chuncheon and were subjected to serology (2-ME RSAT and ICA for B. canis, Rose Bengal test and C-ELISA for B. abortus), blood culture and 3 types of PCRs (BSCP31, 16s rRNA, and OMP-2). All blood samples were negative by serology and blood cultures. The BCSP31 and the OMP-2 PCR detected 5 samples were positive whereas the 16S rRNA PCR detected all samples were negative as serological methods and blood culture did. From the results observed in the present study, we conclude that 16S rRNA PCR could be used for direct PCR for canine blood samples.