• 대한수의학회지
• /
• 제43권1호
• /
• pp.67-75
• /
• 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
• /
• 제37권1호
• /
• pp.45-49
• /
• 2014

Pullorum disease and fowl typhoid are septicemic avian diseases transmitting through egg by transovarian infections. A series of tests has been performing in breeding flocks of chickens and test plans for proper inspection have been modified by government veterinary institute to control of such diseases. To improve inspection plans, different test methods were compared using fowl typhoid positive samples from a poultry farm located in Jeonbuk state in 2012. Based on first inspection, 11 samples among total 200 samples were positive by rapid slide agglutination (RSA) test and 7 samples among RSA positive samples were finally diagnosed as Salmonella Gallinarum infection by ELISA, bacterial isolation, PCR, and histopathologic examination. In the second inspection, 20 samples among total 100 samples were positive by RSA test. Among RSA positive ones, 19 samples were positive by ELISA, S. Gallinarm were successfully isolated in 3 samples, and 16 samples were positive by PCR in the cecal tonsils where were not successful for bacterial isolation. Based on histopathologic examination, severe inflammation in the 13 cecal tonsils and infiltration of lymphocytes and heterophils in the 11 livers were observed. Therefore, we suggest that bacterial isolation, PCR, and histopathologic examination methods in the third inspection need to be further used in various tissues for correct diagnosis and for final eradication of pullorum disease and fowl typhoid in breeding flocks of chickens.

• 한국동물위생학회지
• /
• 제33권1호
• /
• pp.29-35
• /
• 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
• /
• 제34권2호
• /
• pp.159-166
• /
• 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
• /
• 제22권3호
• /
• pp.221-237
• /
• 1999

To investigate the specificity of rapid slide agglutination test for pullorum-gallinarum diseases and to obtain a basic data for avian salmonellosis control, salmonella isolation was peformed for the layer chickens positively reacted in pullonlm-typhoid agglutination test. The biochemical, serological and antimicrobial properties of the isolates were examined. The results obtained through this study were summarized as follows; 1. Of 2,384 chickens tested by the agglutination test, 606 chickens (25.4%) were positive reactors. 154 of 606 reactors and 49 of the non-reacting chickens were investigated for salmonella isolation, resulting in isolation of 68 strains of salmonellae from 27 chickens. 2. By organs, the isolation frequency from liver, cecum, spleen, ovary and gall bladder showed 8.9% (18 strains), 8.9% (18 strains), 7.4% (15 strains), 4.4% (9 strains) and 3.9% (8 strains), respectively. 3. By culture medium the combination of selenite broth and MacConkey agar revealed the highest isolation rate and the enrichment culture by delayed secondary enrichment culture method was found the most effective for salmonella isolation. 4. The serotypes of 68 salmonella isolates were identified as 3 strains of S pullorum, 24 strains of S gallinarum, 15 strains of S typhimurium, 8 strains of S enteritidis, 7 strains of S paratyphi A, 5 strains of S typhimurium and 6 strains of the other salmonellae. 5. The serotypes of 8 salmonella strains isolated from 49 chickens non-reacting in pullorum-typhoid agglutination test were identified as 3 strains of S typhimurium and 5 strains of S infantis. 6. When 24 chickens of which 68 strains of salmonellae isolated were examined by microplate agglutination test, the average antibody titer for pullorum antigen was $2^{5.25}$. The chickens at antibody titer between $2^3$ and $2^5$ showed the higher frequency of isolation as compared with the chickens at the other titers. 7. When salmonella isolates were tested the antimicrobial drug sensitivity by disk diffusion method, S paratyphi A were highly sensitive by 100% to ATM and GM, S typhimurium, by 88% to AM, CIP, IMP and TN, S infantis, by 100% to AM, CRO, ENR and PIP, S enteritidis,by 100% to IMP and PIP, S pullorum, by 100% to ATM, CRO, ENR and PIP and S gallinarum, by 92% to CRO, CIP and PIP.

• 대한수의학회지
• /
• 제39권6호
• /
• pp.1099-1105
• /
• 1999

Reproductive failures, such as abortions were occured among dogs in a large kennel in Chonnam area April in 1994. After an initial positive result by the 2 mercaptoethal rapid slide agglutination test(2ME-RSAT) to Brucella canis on five sera, additional specimens from all dogs in the population were tested. The blood cultures and 3 following serological tests (2ME-RSAT, TAT, AGID) performed on all samples on the basis of surveys, the following results were obtained. 1. Thirty three of 62 dogs were seropositive. 2. Twenty blood samples from 33 dogs were cultured, all of the isolates were identified as B canis. 3. Although there was not siginificant difference in sex, age and breed of the cause brucellosis dectected, abortions was occured late in gestation stage.

• 대한미생물학회지
• /
• 제15권1호
• /
• pp.9-17
• /
• 1980

In the ecology and epidemiologic studies on various serotypes of atypical mycobacteria(AM), Schaefer's bacterial agglutination test(BA) provided the basis of the serologic procedures. Recently, attempts have been made to modify and to simplify the Schaefer's BA such as a slide agglutination test(Engel & Beerwald, 1970), a "simplified" BA(Reznikov & Leggo, 1972), an agglutination inhibition test(Richards & Eacret, 1972) and "micromethod"(Thoen et al., 1975). The BA, however, was not widely applied as a routine laboratory test mainly because it requires much times and labors to perform and partley because it is not applicable to hydrophobic strains either often encountered in the isolation of AM in the clinical bacteriology or stock strains maintained in the laboratory. On the contrary, fluorescent antibody technique with mycobacteria may have advantages over the BA because it is far more simpler in serologic procedures and is applicable to all strains of mycobacteria regardless of smooth or rough types of cultures. At the present, it is well known that the type-specific antigens are lacking on the surface of rough type of AM compared to that on smooth type of strain, but the antigenicity on the surface of the hydrophobic strains of AM which resulted from a series of subculture and the strain in the laboratory for 3 to 6 months has not been clarified. In this study, an attempt to serotype the hydrophobic strains of M. scrofulaceum serotype 41, 42 and 43 by fluorescent anti-complement(FAC) technique was made. The FAC technique with mycobacteria was also described in detail. In the summary, the complement fixing antibody titres of reference sera to smooth types of homologous serotype was highest, but the antibody titres of reference sera to hydrophobic strains of serotypes, 41, 42 and 43 gave two-to 8-folds lower than those to smooth type of strains. Although the sensitivity of type-specific antigens on the hydrophobic strains to reference sera was much lower, using the two units of reference sera determined by titration with hydrophobic strains, three serotypes, i. e., 41, 42 and 43 were specifically differentiated one another by FAC technique. This result indicated that the hydrophobic strains which were maintained in the laboratory at least for 6 months still retain type-specific antigen detectable by FAC technique.

• 한국동물위생학회지
• /
• 제20권1호
• /
• pp.69-77
• /
• 1997

To invastigate the epidemiological trait of listeriosis, Listeria monocytogenes were isolated from the carcasses of pigs and cattle, and environmental specimens in slaughter house. Also serotype of isolates were classified by rapid slide agglutination test. In the carcasses of pigs, Listeria sp were isolated from the carcasses after bleeding(62%), after dismemberment(60.0%) and before shipping(76.0%), and L monocytogenes were present in 8% of the carcasses after dismemberment and in 14% of the carcasses before shipping. However, few Listeria sp were isolated from the living body skin and the carcasses after scalding. In the carcasses of cattle, Listeria sp were isolated from the carcasses after bleeding(10%), after dismemberment(36.7%) and before shipping(63.3%), L monocytogenes were present in 3.3% of the carcasses after dismemberment and in 10% of the carcasses before shipping. Overall, L monocytogenes, L innocua, L welshimen, L grayi, and L murrayi were present In 4.8, 40, 2.3, 2.6 and 0.3% of all the carcasses, respectively. Prevalence of Listeria sp in environmental specimens were found to be 80% in slaughter house floors and 100% in sewage, and L monocytogenes were present in 15% of sewage. However, few Listeria sp were isolated from chilled water and from scalding water. Overall, L monocytogenes, L innocua, and L welshimeri were present in 3.8, 45 and 6.3% of all the environmental specimens, respectively. A total 27 strains of L monocytogenes were isolated from samples tested and all of the strains were classified into serotype 1.

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