肉鸡和蛋鸡鼻气管鸟杆菌的分离鉴定及其新生物学特性分析

Isolation,identification and the novel biological characterization of Ornithobacterium rhinotracheale from layers and broilers

为了确诊疑似肉鸡和蛋鸡鼻气管鸟杆菌（0RT）感染并研究其病原的新生物学特性，对其病原进行分离鉴定并分析其16SrDNA序列特征、溶血活性、凝血活性和药物敏感性。从云南省有鼻炎症状的蛋鸡和肉鸡中分离到5株革兰氏阴性多形性杆菌，分别被命名为YNll061、YN12071、YN12072、YN12091和YN12092。生化试验表明，5株分离株氧化酶阳性，过氧化氢酶阴性，发酵葡萄糖、乳糖、半糖、麦芽糖、甘露糖和果糖，产酸不产气；16SrDNA序列分析表明，5株分离株与GenBank上12株国外代表株之间的核苷酸序列同源性为99．39／6～100％，进一步鉴定为ORT。溶血试验和凝血试验表明，5株分离株具有p溶血性，能够凝集戊二醛醛化的绵羊红细胞。5株分离株同时对链霉素、庆大霉素、卡那霉素、阿米卡星、红霉素、氧氟沙星、诺氟沙星显示多重耐药性。结果证实，云南省存在ORT感染。本研究首次报道了ORT的溶血性和凝血性，为ORT的分离鉴定和新型诊断检测方法的建立奠定了基础。

To diagnose a suspected Ornithobacterium rhinotracheale （ORT） infection and study the novel biological characterization of ORT, isolation, identification, sequence of 16 S rDNA gene, hemaggluti- nating activity,hemolytic activity and antibiotic susceptibility of ORT were studied. Five isolates of Gram- negative, polymorphic and rod-shaped bacteria, named as YNl1061, YN12071, YN12072, YN12091 and YN12092,were isolated from the layers and broilers with coryza-like respiratory disease in Yunnan Pro- vince. The 5 isolates were oxidase-positive and catalase negative,and were able to ferment glucose,lactose, galactose,maltose,mannose and fructose to produce acid but no gas. The bacteria were further identified as ORT by 16 S rDNA sequence analysis,which shared identity from 99.3% to 100% between the 5 isolates and 12 representative isolates from GenBank. Hemolytic and hemagglutinating tests showed that these 5 i- solates had/9 hemolytic activity,and were able to hemagglutinate glutaraldehyde-fixed sheep erythrocytes. Antibiotics susceptibility test showed that the isolates were of 1（50 ~ multiple antibiotic resistance to strep- tomycin, gentamicin, kanamycin amikacin, erythrocin, ofloxacin, norfloxacin. This study confirmed that the presence of ORT infection in broilers and layers in southwestern China. It is also the first report of hemag- glutinating activity and hemolytic activity of Chinese ORT isolates, and this finding will contribute to the identification and development of new detection methods for ORT.