Animal Health Department, Veterinary School, Universidad Complutense de Madrid, Spain.
Animal Health Department, Veterinary School, Universidad Complutense de Madrid, Spain.
Vet Microbiol. 2019 May;232:70-73. doi: 10.1016/j.vetmic.2019.04.006. Epub 2019 Apr 6.
The P. multocida toxin (PMT), a dermonecrotic protein encoded by the toxA gene, is the major virulence factor of capsular type D P. multocida strains causing progressive atrophic rhinitis (PAR) in pigs. A high frequency of P. multocida isolates harboring the toxA gene has been found among ovine pneumonic isolates, although the ability of these isolates to express PMT has never been examined. In this study we have investigated the ability of ovine toxA+ P. multocida isolates (n = 57) to express a functional toxin by detection of PMT toxin antigen using an ELISA test and its cytopathic effect in a Vero cell assay. PMT antigen was expressed in the great majority (54/57; 94.7%) of toxA+ isolates. Moreover, the 100% toxA+ ovine isolates analyzed produced a cytopathic effect in Vero cells within 24-48 h post-inoculation, identical to that described for porcine toxigenic P. multocida isolates. These results show for the first time that, in addition to isolates associated with PAR, isolates of P. multocida associated with pneumonia in sheep are also toxigenic. In addition, we found a total agreement (Kappa = 1; C.I. 0.75-1.25) between the detection of the toxA gene and the toxigenic capability of P. multocida isolates, indicating the PCR detection of toxA would be a suitable predictive marker of the toxigenic fitness of P. multocida.
多杀性巴氏杆菌毒素(PMT)是一种由 toxA 基因编码的皮肤坏死蛋白,是导致猪进行性萎缩性鼻炎(PAR)的荚膜 D 型多杀性巴氏杆菌菌株的主要毒力因子。在绵羊肺炎分离株中发现了携带 toxA 基因的多杀性巴氏杆菌分离株的高频率,尽管这些分离株表达 PMT 的能力从未被检测过。在这项研究中,我们通过 ELISA 试验检测 PMT 毒素抗原,并用 Vero 细胞试验检测其细胞病变效应,研究了绵羊 toxA+多杀性巴氏杆菌分离株(n=57)表达功能性毒素的能力。在大多数(54/57;94.7%)toxA+分离株中表达了 PMT 抗原。此外,分析的 100% toxA+绵羊分离株在接种后 24-48 小时内会在 Vero 细胞中产生细胞病变效应,与猪产毒多杀性巴氏杆菌分离株相同。这些结果首次表明,除了与 PAR 相关的分离株外,与绵羊肺炎相关的多杀性巴氏杆菌分离株也是产毒的。此外,我们发现 toxA 基因的检测与多杀性巴氏杆菌分离株的产毒能力之间存在完全一致(Kappa=1;置信区间 0.75-1.25),表明 PCR 检测 toxA 可能是多杀性巴氏杆菌产毒适应性的合适预测标志物。