Varinrak Thanya, Muenthaisong Anucha, Apinda Nisachon, Sawada Takuo, Sthitmatee Nattawooti
a Faculty of Veterinary Medicine , Chiang Mai University , Muang, Chiang Mai , Thailand.
b Laboratory of Veterinary Microbiology , Nippon Veterinary and Life Science University , Musashino, Tokyo , Japan.
Avian Pathol. 2019 Feb;48(1):4-11. doi: 10.1080/03079457.2018.1533239. Epub 2018 Oct 22.
A capsule-defective mutant strain PBA129 of Pasteurella multocida was constructed by electroporation of phagemid containing the coding region of the antisense RNA of the ompH gene into the wild type strain X-73 (serovar A:1) of P. multocida. The pathogenicity and protective potency of the mutant against homologous and heterologous challenge in mice and chickens were characterized. Greyish colonies of the mutant, indicating lower capsule thickness, on selective dextrose starch agar were observed under an obliquely transmitted light stereomicroscope and compared to iridescent colonies of the wild type strain X-73. Strain PBA129 had lower capsule thickness than the wild type strain as observed with an electron microscope. Strain PBA129 was apparently attenuated, as mice and chickens inoculated with the bacteria at 10 CFU survived. Protection was observed in both mice and chickens inoculated with strain PBA129 upon challenge exposure to avian P. multocida strains X-73 and P-1059 (serovar A:3), respectively. In conclusion, the mutant strain PBA129 of P. multocida strain X-73 was completely attenuated, and it was possible to induce sufficient protection against avian P. multocida strains.
通过将含有ompH基因反义RNA编码区的噬菌粒电穿孔导入多杀性巴氏杆菌野生型菌株X-73(血清型A:1),构建了一株多杀性巴氏杆菌的荚膜缺陷突变株PBA129。对该突变株在小鼠和鸡体内针对同源和异源攻击的致病性和保护效力进行了表征。在斜射光立体显微镜下观察到突变株在选择性葡萄糖淀粉琼脂上的菌落呈灰白色,表明荚膜厚度较低,并与野生型菌株X-73的彩虹色菌落进行了比较。用电子显微镜观察发现,菌株PBA129的荚膜厚度低于野生型菌株。菌株PBA129明显减毒,因为接种10 CFU该细菌的小鼠和鸡存活了下来。分别用菌株PBA129接种小鼠和鸡,在受到禽多杀性巴氏杆菌菌株X-73和P-1059(血清型A:3)攻击时,观察到了保护作用。总之,多杀性巴氏杆菌菌株X-73的突变株PBA129完全减毒,并且有可能诱导对禽多杀性巴氏杆菌菌株的充分保护。