• Korean Journal of Veterinary Service
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• v.35 no.4
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• pp.275-281
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• 2012

Canine brucellosis is the zoonosis in worldwide and Brucella (B.) canis is a facultative intracellular pathogen that has a very limited host. MLVA-16 (Multilocus VNTR analysis) is a efficient method for genotyping of Brucella species. Various methods have been established for genotyping of Brucella species, but most of analytical method is lack reproducibility and limited capability to differentiate them. B. canis isolates (n=73) from 7 farms in Gyeongbuk province in 2003~2010 were analyzed using 16 VNTR loci. Automatic electrophoresis system was utilized for more high throughput and rapid simple discrimination. Thirty two genotypes were identified from 73 B. canis isolates. MLVA could contribute to molecular typing for epidemiological evaluation of canine brucellosis.

• Korean Journal of Veterinary Service
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• v.29 no.1
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• pp.47-53
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• 2006

For examination of antibiotic therapeutic efficacy in canine brucellosis, this examination was carried out two female bitches infected with Brucella canis in Gyeongbuk province, and used combicillin, baytril and doxycycline in susceptible antibiotics at B canis. During 18 month after the termination of antibiotic therapy, blood sample of the two bitches were examined for B canis antibody and antigen. The antibody of one bitch was disappeared at 5 month after antibiotic therapy and the other was continued at 18 month, but two bitches were not detected antigen by blood culture and PCR. Examination of blood chemical value (AST, ALT, urea, creatinine) of two bitches was increased in AST value during antibiotic therapy.

• Korean Journal of Veterinary Research
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• v.41 no.1
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• pp.67-71
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• 2001

A bacteriological and serological investigation was conducted in a commercial breeding kennel in Taegu city in which canine abortion caused by Brucella canis occurred, and the family dogs in this area were also surveyed for Br canis infection during a period from March 1999 to May 2000. Of 195 dogs in the breeding kennel, 50(25.6%) were found to be bacteremic and 82(42.1%) were shown to be positive for canine brucellosis by both bacteriological and serological test. Of 357 family dogs examined, 17(4.8%) had an agglutinin titer of 1 : 160 or higher. Of these 17 dogs, 5(2.8%) were from indoor dogs and 12(6.7%) were from outdoor dogs. Only 2 mongrel dogs(1 female and 1 male) of 17 serologically positive dogs showed an agglutinin titer of 1 : 2560 or higher and Br canis was isolated from their blood.

• Korean Journal of Veterinary Service
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• v.30 no.3
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• pp.363-374
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• 2007

A biochemical characterization and antimicrobial drugs susceptibility study was conducted in four breeding kennel which was canine abortion caused by Brucella canis in Gyeongbuk province in 2003-2006. Total of 267 dogs domesticated in the four kennel were examination. Among them, 143 (53.6%) dogs were sero-positive and 25 of blood samples were isolated to Brucella canis. At amplification of 35KDa-BCSP gene using PCR, 711 bp DNA fragment was same visible in 25 isolates and B canis RM6/66. Biochemical characterization of B canis isolated was non-hemolytic, no production of $H_2S$, no fermentation of carbohydrates, catalase-positive, oxidase-positive, indol-negative, hydrolyzation of urea, reduction of nitrate and development of thionin dye medium. Using disk-diffusion method, all of 25 strains tested were found to be highly susceptible to tetracycline, aminoglycoside, quinolone, macrolide antibiotics, rifampin and ampicillin in vitro. Using PFGE with restriction enzyme Smi I, 25 isolates tested were typed to 2 pattern, S1 and S2.

• Korean Journal of Veterinary Research
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• v.43 no.1
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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.

• Korean Journal of Veterinary Research
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• v.49 no.2
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• pp.105-111
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• 2009

In this study, occurrence of canine brucellosis was surveyed in kennels, indoor dogs and stray dogs in Korea, and infrequent restriction site-polymerase chain reaction (IRS-PCR) was applied to analyze DNA polymorphism of Brucella canis (B. canis) isolates. Among a total of 501 dogs tested, B. canis antibodies by both rapid screening agglutination with 2-mercaptoethanol (2-ME RSAT) and immunochromatographic assay were detected in only 14.1% of kennel dogs. There were no seropositive cases in indoor dogs and stray dogs. DNA polymorphism was observed in 16 B. canis isolates by the IRS-PCR. Sixteen isolates were tested with primers, PsalA, PsalC, PsalG and PsalT, and different primers produced different DNA patterns. In regard to the IRS-PCR pattern of 16 isolates, 9 (56.3%) belonged to the IRS-PCR type I. The remaining 7 were differentiated as type II, III and IV. An application of the primer PsalC provided discrimination between B. canis isolated in 2005 and others.

• Korean Journal of Veterinary Service
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• v.33 no.1
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• pp.23-27
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• 2010

This survey was performed to investigate the seroprevalence of antibodies to Brucella canis in dogs from public animal shelters and breeding kennels in Incheon. A total of 402 dogs selected randomly were tested serologically by using immunochromatographic antibody test kit. None of 289 dogs in public animal shelters were sero-positive and 10 (8.9%) of 113 dogs in breeding kennels were sero-positive. 5 (4.4%) strains of Brucella canis were isolated from 10 sero-positive dogs' blood. Antimicrobial susceptibility test was carried out by Disk diffusion method. They were susceptible to tetracyclines, quinolones, aminoglycosides and combination amoxicillin with clavulanic acid.

• Korean Journal of Veterinary Research
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• v.39 no.6
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• pp.1106-1111
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• 1999

In a kenel of 62 dogs, 33 were diagnosed as infected dogs Brucella canis by serological test and blood cultures in a large kennel in Chonnam area. Twenty eight of 33 dogs were treated with combined antibiotics therapy consisting of tetracycline and dihydrostregtomycin. After the first treatment, all dogs became abacteremic and serologic titers declined. Abortion due to B canis infection could be prevented by antibiotic therapy during pregnancy. However, sequential antibiotics therapy for 4 weeks did not eradicate B canis from affected bitch. According to results of serological and blood culture tests, effect of antibiotics treatment respectively revealed that 17 of the originally infected dogs were cure with second therapy schedule at 2 months and 27 of that with 6 months after second therapy.

• Korean Journal of Veterinary Service
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• v.33 no.1
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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.

• Korean Journal of Veterinary Service
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• v.34 no.2
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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.