• 대한수의학회지
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• 제10권1호
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• pp.11-23
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• 1970

Recently Carter(1952) reported the capsule antigens of Pasteurella multocida could be divided into four serological types A,B,C and D by means of precipitation tests. Subsequently he showed that the most sensitive for identification of these types involved the use of capsule substance adsorbed by erythrocytes in hemagglutination test. It may be somewhat difficult to conduct the hemagglutination test in small laboratory, because relatively large amounts of antisera and erythrocytes of the human O type are required for the test. A simple method for serological typing of P. multocida was the slide agglutination test employed by Little et al. (1943) and Namioka et al. (1962), but this method is still in controversy. The author tried adapting Carter's hemagglutination method to the slide method so called "micromethod technique", and studied on the stabilization of erythrocytes for use of slide hemagglutination to P. multocida although many invesigators reported the stabilization of erythrocytes. The results obtained are summarized as follows: 1. A simplified method (slide method) for capsule typing of the organism was developed by adapting Carter's hemagglutination reaction(tube method). Antibody-containing serum can be diluted serially on Boerner's microtest slide with capillary or serological pipetts with a considerable accuracy. The slide reaction can be carried out with case on the slide by adding $0.05m{\ell}$ of antigen-sensitized erythrocytes suspension diluted to one percent on $0.05m{\ell}$ of serially diluted antibody-containing sera, and the final result can be read after 60 minutes at the room temperature ($15^{\circ}C$). 2. It is difficult to determine superiority of inferiority between the slide method and the tube method on the pattern of the reaction of hemagglutination. 3. The pH range of 6.6 to 8.3 is optimal for the slide hemagglutination reaction. 4. The antigen-sensitization against erythrocytes at $37^{\circ}C$ is optimal for the slide hemagglutination. 5. Both the doses and concentration of antigen do not influence the antigen-adsorbing capacity of erythrocytes. 6. The reduction of antigen-sensitizing hours does not influence the antigen-adsorbing capacity of erythrocytes even 30 minutes. 7. The tannic acid treatment against formalinized and non-formalinized erythrocytes showed no effect on the reaction of hemagglutination. 8. The erythrocytes preserved at $4^{\circ}C$ in the ACD solution do not decrease the reactivity on the reaction of hemagglutination for 60 days, while they begin slight hemolysis 30 days after preserving. 9. The stable preparation of erythrocytes can be obtained by treating the cells at $37^{\circ}C$ for 20 hours with from 4 to 8 percent of formalin in saline or buffer. These cells can be preserved at $4^{\circ}C$ for more than 8 months experimented without hemolysis. With low concentration of formalin, the cells were not sufficiently stabilized resulting in the hemolysis after short period of preservation at $4^{\circ}C$. 10. The erythrocytes treated with 16 percent of formalin remain constantly or increase the reactivity for the reaction of hemagglutination. On the contrary, the cells treated with I to 8 percent of formalin decrease the reactivity. 11. There is no difference between nontreated fresh erythrocytes and the erythrocytes preserved in the ACD solution on the reactivity against the hemagglutination, and the erythrocytes treated with 16 percent of formalin showed the reactivity of higher level than that of the above two kinds of erythrocytes. 12. There is no difference between the saline and the isotonic buffer solution on the reaction of hemagglutination.

• 한국동물위생학회지
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• 제31권3호
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• pp.347-356
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• 2008

The purpose of this study was to evaluate the efficacy and cross-protection of serovar 12 against serovar 4 or 5 of H parasuis with M+$Parapac^{(R)}$. A total of 141 piglets from 2(A and B) farms were used and divided into experimental group and control group in each farm. Farm A has been detected H parasuis serovar 12, whereas farm B has been detected H parasuis serovar 4 or 5 from post-weaned pigs with PMWS. The piglets were vaccinated intramuscularly with 2.0ml of M+$Parapac^{(R)}$ in experimental group or normal saline in control group at 1 week of age. A same booster dose was given at 3 weeks of age. In order to value the antibody titer to H parasuis using by tube agglutination test, blood samples were collected from piglets at the aged of 1 week, 6 and 14 weeks. In experimental group and control group, the average antibody titers were $32.5{\pm}21.0,\;114.5{\pm}34.0,\;98.1{\pm}55.4$ and $32.9{\pm}18.6,\;25.8{\pm}36.9,\; 746.7{\pm}1,215.8$ at the aged of 1 week, 6 and 14 weeks, respectively. The cumulative clinical sign indexes by standard guideline of Schering-Plough Animal Health Corp were 486 and 1,069, respectively. The average daily gains and feed conversion rates were $0.553{\pm}0.016kg$ and $0.492{\pm}0.004kg$, and 1.99 and 2.24, respectively. The average gross lesion scores were $1.0{\pm}0.8$ and $1.9{\pm}0.6$, respectively. According to these results, the M+$Parapac^{(R)}$ containing H parasuis serovar 12 may be induce circulating antibodies that cross-react with serovar 4 or 5 and have a protection of PMWS with H parasuis.

• 한국동물위생학회지
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• 제18권2호
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• pp.133-151
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• 1995

During 2 years from Octorber 1992 to April 1994, prevalence of general respiratory diseases and atrophic rhinitis in the pig herds located in the Western Chungnam was investigated, and isolation of B. bronchiseptica was attempted for the pigs manifested with the clinical signs of atrophic rhinitis(AR). The isolates were characterized and identified in aspects of biochemical properties, antigenicity, drug sensitivity and pathogenicity. The results obtained through the experiments are summarized as follows; 1. During 2 years of investigation, the overall prevalence of the general respiratory diseases in the pi8 herds in Western Chungnam was 35.3%, consisting of 35.1% in the pig farms and 38.8% in a slaughter house. The prevalence by age groups accounts for 9.2% in adults, 44.7% in rearings and 25.3% in sucklings. By farm size, The highest prevalence of 56.5% was observed in the smallest farm with 1 to 200 heads. 2. The prevalence of clinical cases of artrophic rhinitis was recorded by 12.7% in the group that is the sows and piglets vaccinated, 28.9% in the group that is the sows only vaccinated and 39.8% in the group of the non-vaccinated groups. In the slaughter house, 53(24.8%) of 214 pigs examined exhibit the AR lesions. 3. A total of 189 strains of B. bronchiseptica were isolated from the pig herds. Isolation rates were 12.6% in the group that is the sows and piglets vaccinated, 34.1% in the group that is the sows only vaccinated and 45.7% in the group of the non-vaccinated groups. Isolation rate in the specimen from the slaughter house was 93( 43.5% ) of 214 pigs examined. Of the AR-non-vaccinated group, the piglets aged bet- ween 61 to 90 days revealed the highest isolation rate of 58.5%. 4. The titers of antibody against B. bronchiseptica were measured by tube agglutination test. The group that is the sow and piglet-vaccinated showed the highest titer of 640-2, 560 in sow and 640longrightarrow5, 120 in piglet. The group that is the sows only-vaccinated revealed 640-2, 560 in sows and 640-1, 280 in piglets. Both of the vaccinated groups showed 100% positive reaction. The group of the non-vaccinated sho-wed relatively lower titer of 0-1, 280 in both of sows and piglets. The positive rate of the sera obtained from the slaughter house was 53.3% with the antibody titer of 0-1, 280. 5. Biochemical and serological properities of 189 isolates were very similar to those of the reference B. bronchiseptical phase I type, indicating that most of isolates are B. bronchiseptica phase I type. 6. In antimicrobial drug susceptibility, 87.3% of 189 isolates was susceptible to chloramphenicol, 79.9%, to amikacin, 64.6%, to cephalothin and less than 35.4% to others. 7. In agar-gel immunodiffusion and SDS-PAGE analysis, the isolates presented the identical antigenicity and protein profiles to the reference standard strains. 8. The whole cells and bacterial filtrates of the isolates were inoculated to guinea pigs and mice. The isolates showed the hish pathogenicity and dermonecrotoxiciy.

• Journal of Preventive Medicine and Public Health
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• 제42권4호
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• pp.237-242
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• 2009

Objectives : The incidence of zoonoses in Korea has increased recently. However, the study of high risk groups for zoonoses has not been conducted to date in Korea. Thus, we did this study to obtain data on brucellosis among slaughterhouse workers in Korea. Methods : We evaluated the structure of slaughterhouses and the process of slaughtering by reviewing the relevant literature and doing field studies. We visited 73 slaughterhouses and 62 residual products handle houses across the country. In addition, we conducted a questionnaire survey of the work activities, and obtained blood samples in order to determine the seroprevalence and risk factors of brucellosis. The titers of brucellosis antibodies were measured using the standard tube agglutination test SAT). We diagnosed subjects as seropositive for Brucellosis if the titers were more than 1:160. The data collected was evaluated using SPSS ver. 17.0. Results : We included 1,503 subjects and obtained 1,482 blood samples among them: 849 workers involved in slaughtering, 351 handlers of residual products, 190 inspectors and their assistants, and 92 grading testers and their assistants. The seroprevalence of brucellosis among the slaughterhouse workers was 0.8% (95% CI=0.4-1.5). Broken down, the seroprevalence of brucellosis among the workers involved in slaughtering was 0.7% (95% CI=0.3-1.6), the handlers of residual products was 1.7% (95% CI=0.7-3.9) respectively. Risk factors for contracting brucellosis among slaughterhouse workers were being splashed with cattle blood around the mouth, cattle secretions around the body and not putting on protective apron while at work. Conclusions : An educational program is needed for high risk groups on zoonoses about the prevention of infection. Thus, effective working guidelines for workers who participate in the slaughter of animals must be developed in order to protect them from zoonoses.

• 대한수의학회지
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• 제44권3호
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• pp.433-439
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• 2004

This study was carried out to investigate the efficacy of rifampin with or without streptomycin in male Sprague-Dawley (SD) rats experimentally inoculated with Brucella abortus. Thirty rats were intraperitoneally inoculated with $1.0{\times}10^9$colony-forming units of B. abortus. They were divided into 3 groups by treatment with antibiotic. 10 rats in Group A were orally administrated with rifampin, 10 rats in Group B with rifampin orally and with streptomycin intramuscularly over 12 weeks starting at 1 week post infection (PI). A placebo recipient in Group C was inoculated with sterile saline without antibiotics. All animals were monitored by tube agglutination test (TAT) and AMOS-PCR to evaluate the efficiency of the antibiotics to B. abortus infection. The antibody titers in Groups A, B and C were 1:400, 1:400 and 1:800 as measured by TAT at the first week PI, respectively. The antibody titer in Group A decreased to 1:100 by the 13th week PI. That in the control group was observed as high antibody titer until 13th weeks PI, but the antibody response in Group B was low(1:50) from the 5th week to the 13th week PI. AMOS-PCR there was evidence of relapse of B. abortus in group A in liver and spleen specimens at the 13th week PI. B. abortus DNA was detected in Group C in liver and spleen specimens from the 1st week to 13th week PI by AMOS-PCR. However AMOS-PCR could not detect any organism in Group B from the 3rd week PI until the end of the study. This study demonstrated that administration of a combination of rifampin and streptomycin was more efficacious than administration of rifampin alone. A significant reduction in antibody titer was observed when a combination of 15 mg/kg/day of rifampin per os and 15 mg/kg/day streptomycin intramuscularly was used in comparison with the antibody of control group.