忠南地域 屠畜豚의 肺病變으로부터 分離한 Actinobacillus pleuropneumoniae의 生物學的 및 免疫學的 特性 = Studies on biochemical and immunological properties of Actinobacillus pleuropneumoniae isolated from the slaughter pigs with respiratory lesions in Chungnam province

다국어 초록 (Multilingual Abstract)

An epidemiologic study on pleuropneumonia in the slaughter pigs(Chonan and Asan area, Chungnam province, Korea) during the period of January 1994 through December 1995 was conducted. Isolation of A. pleuropneumoniae was attempted in 425 pigs with pneumonic lesions. Biochemical properties, antimicrobial susceptibility, serotypes and pathogenicity of A. pleuropneumoniae isolates were investigated. In addition, outer membrane protein(OMP) of the isolates were extracted to determine its properties and immunogenicity in both mice and piglets. Of 3,395 slaughter pigs, pleuropneumonia was observed in 425 pigs(10.6%). A. pleuropneumoniae was isolated from 22 pigs(5.2%) out of 425 pigs with pneumonic lesions. The biochemical properties of all isolates were same as those of reference A. pleuropneumoniae strain. Among 22 isolates, 9, 1 and 12 isolates were serovar 2, 3 and 5, respectively. The results of antimicrobial susceptibility test revealed that the isolates showed high susceptibility to ciprofloxacin and cephalothin, moderate susceptibility to amikacin, gentamicin, kanamycin and streptomycin, and low susceptibility to erythromycin, tylosin and sulfadimethoxin. The isolates were varied in pathogenicity to mice. Median lethal dose of LE9402(serovar 2) and LE9511(serovar 5) were 9.2×10^7CFU and 2.8×10^7CFU, respectively. Specific pneumonic lesions were observed from the infected mice with clinical signs. Bacteria recovery rate was high in the lung, but low in heart blood and tracheas. Serovar 2 was found to be more pathogenic than serovar 5 in guinea pig. Mortality on guinea pigs inoculated with serovar 2(5.4×10^8 - 5.4×10^6CFU) abd serovar 5(2.8×10^8 - 2.8×10^6CFU) was 20-40% and 40-80%, respectively. A severe hemorrhagic lesions and focal pneumonic lesions were observed from dead guinea pigs. Bacteria recovery rate was relatively higher in the lung than that of other organs. In the SDS-PAGE analysis, OMP-enriched fractions of both isolates and reference strains contain common peptide bands equivalent to molecular weight of 17, 27, 42, 52 and 95Kd. In addition to common peptide bands, the bands which are specific to each isolate were also observed. The profiles of Sephadex G25 fractions showed 3 major peaks. The common peptide bands which were observed by SDS-PAGE of the crude OMPs were found i the peaks 1 and 2. The OMPs extracted from serovar 2(LE9402) and serovar 5(LE9511) provided high level of protection in mice(70-80%) and pigs(100%). All animals inoculated with OMPs were seroconverted, showing micro-agglutination titer of 640 to 1280.