鸭疫里氏杆菌形态及超微结构与外膜蛋白在抗血清诱导下的改变  

Changes of morphology,ultrastructure and the outer membrane proteins of Riemerella anatipestifer grown in medium with its duck antisera

作　　者：高继业[1] 崔兆莲 黄伟[1] 杨致邦[2] 王振兴[1] 温亚兰 范贵荣[2] 

机构地区：[1]西南大学荣昌校区,重庆402460 [2]重庆医科大学基础医学实验教学中心病原生物学与免疫学实验室,重庆400016

出　　处：《中国兽医科学》2013年第11期1109-1114,共6页Chinese Veterinary Science

基　　金：中央高校基本科研业务费专项(XDJK2013C009)

摘　　要：为研究鸭疫里氏杆菌(Riemerella anatipestifer)感染康复鸭抗血清的体外抗菌活性,将血清2型R.anatipestifer AF株接种于含抗血清的TSB培养基并培养10h,在电子显微镜下观察其细胞形态和超微结构的变化,并分别采用SDS-PAGE和RT-PCR技术对其OMPs成分和OmpA表达水平进行分析。结果显示,R.anatipestifer菌体细胞由杆状变为中部有环形凹陷的球状,活菌细胞胞内及胞间有大量外膜泡,且部分细胞胞质内及胞内膜泡边缘分布有电子密度较高的稠密小体;另外,菌体细胞的OMPs较阴性对照组和空白对照组缺少6条蛋白多肽,分子质量大小分别为33、36、44、50、55和65ku,且OmpA的表达量显著下降(P<0.01)。研究结果证明,R.anatipestifer在中部及端部存在3个潜在的分裂位点,抗血清能诱导分裂位点发生改变并抑制OmpA的表达,致使其形态由杆状变为球状。The antibacterial activity of the recovered sera of ducks infected with Riemerella anatiDes- tifer was studied in vitro. AF strain of R. anatipestifer serotype 2 was cultivated for 10 h in TSB medium with the recovered sera,its morphology and ultrastructure were observed by scanning electron microscopy and transmission electron microscopy,and the expression level of OMPs and OmpA were analyzed by SDS- PAGE and RT-PCR,respectively. The morphology of R. anatipestifer cultivated with the recovered serum changed from short rods in normal condition to spheres, which appeared with a annular concavity at the central,and a large number of outer membrane vesicles appeared intracellularly and intercellularly. Some higher electron density dense bodies were also found and distributed in cells or at the edge of outer mem- brane vesicles. Total of six protein bands,such as 33 ku,36 ku,44 ku,50 ku,55 ku and 65 ku were not detec- ted in the main OMPs sample from R. anatipestifer cultivated with the recovered serum compared with the negative serum and no serum. The OmpA expression level decreased significantly in the R. anatipestifer cultivated with the recovered serum（P〈0.01）. It was suggested that R. anatipestifer might be 3 potential division sites which were distributed in its middle and both ends,and that the antibody against R. anatipestifer in the reco- vered duck serum could be able to induce these site change,which inhibit the expression of OmpA.

关 键 词：鸭疫里默氏杆菌 形态学 超微结构 外膜蛋白 

分 类 号：S852.612[农业科学—基础兽医学]