兔溶血性葡萄球菌的分离鉴定及耐药性分析  

Isolation, identification and drug resistance analysis of rabbit haemolytic Staphylococcus

作　　者：伍思华[1] 邬向东[1] 黄靖文 李欢 区海蕴 周亚南 朱芝秀[1] WU Sihua;WU Xiangdong;HUANG Jingwen;LI Huan;OU Haiyun;ZHOU Yanan;ZHU Zhixiu(College of Animal Science and Technology,Jiangxi Agricultural University,Nanchang 330045,China)

机构地区：[1]江西农业大学动物科学技术学院,南昌330045

出　　处：《黑龙江畜牧兽医》2021年第19期96-101,153,共7页Heilongjiang Animal Science And veterinary Medicine

基　　金：江西省教育厅项目(GJJ150416);江西省科技厅项目(20122BBF60082)。

摘　　要：为了了解江西农业大学动物科学技术学院实验动物房试验兔暴发猝死性疾病的原因,试验对死亡兔进行剖检,针对肝脏等典型病变部位进行细菌分离。观察分离菌的菌落形态、进行革兰氏染色镜检,并对小白鼠皮下注射分离菌进行攻毒试验。通过生化试验和16S rRNA测序进一步鉴定细菌种类。根据药敏试验和耐药基因检测进一步分析其耐药特性,应用兔肠乳杆菌发酵液对分离菌进行体外抑菌试验。结果表明:剖检可见呼吸道感染,内脏出血坏死和纤维素性渗出。从死亡兔中共分离到6株细菌,分别命名为TP-1~TP-6。镜检为葡萄状聚集的革兰氏阳性球菌,攻毒试验证明分离菌具有较强毒力。生化试验符合葡萄球菌属特征,16S rRNA测序鉴定6株菌均为金黄色葡萄球菌(S.aureus)。6株菌均为多重耐药菌,TP-4、TP-5、TP-6的Tet(K)耐药基因为阳性。兔肠乳杆菌发酵液对6株S.aureus均具有较好的抑菌活性。说明此次兔暴发的疾病是由高致病型S.aureus感染引起的,该菌具有多重耐药性,兔肠乳杆菌发酵液配合高敏药物是一种不错的治疗方法。In order to understand the cause of the sudden death disease in experimental rabbits in the Experimental Animal Room of the College of Animal Science and Technology of Jiangxi Agricultural University, necropsy was performed on the dead rabbits, and bacteria isolation was carried out from typical lesions such as the liver. Observation of colony shape and Gram-staining microscopy were performed on the isolated bacteria;and bacteria challenge test in mice was performed via subcutaneous injection. Biochemical identification and 16 S rRNA sequencing were used to further identify bacterial species. According to the drug susceptibility test and drug resistance gene detection, the drug resistance characteristics were further analyzed;and the lactic acid bacteria fermentation product was used to conduct an in vitro antibacterial test on the isolated bacteria. The results showed that the necropsy showed respiratory infection, visceral hemorrhage and necrosis and fibrinous exudation. Six strains of bacteria were isolated from dead rabbits and were named TP-1 to TP-6. Microscopic examination showed a grape-like cluster of Gram-positive cocci;and the mouse challenge test proved that the isolate had strong virulence. The biochemical test was consistent with the characteristics of Staphylococcus;and identified six strains were all identified as Staphylococcus aureus(S. aureus) by 16 S rRNA sequencing. The six strains were all multi-drug resistant, and three strains of TP-4,TP-5 and TP-6 were positive for the Tet(K) resistance gene. Lactobacillus fermented product had good antibacterial activity against six strains of S. aureus. The results suggested that the disease outbreak in rabbits was caused by the highly pathogenic S. aureus infection, which had multi-drug resistance. The use of lactic acid bacteria fermentation products combined with highly sensitive drugs was a good treatment method.

关 键 词：兔 剖检 16S rRNA鉴定 药敏试验 耐药基因 抗生素替代 

分 类 号：S852.611[农业科学—基础兽医学] S858.291[农业科学—兽医学]