新疆乌鲁木齐本地某养殖场锦鲤(Cyprinus carpio)突然大批死亡,在排除寄生虫感染后,怀疑是由细菌引起。为明确引起锦鲤大批死亡的致病菌,本实验对病鱼肝脏、肠道、肾脏等组织进行细菌分离纯化。通过革兰氏染色、微量生理生化反应及分子生物学鉴定,成功分离出两株菌,命名为CK5和CK9。两株菌都为革兰氏阴性短杆菌,在30℃正常生长,能利用葡萄糖、乳糖、纤维二糖,并且V-P、明胶、水杨苷实验阳性;PCR扩增其gyr B基因并测序,BLAST比对后,两株菌与维氏气单胞菌(Aeromonas veronii)的同源性最高,分别为99.6%和99.0%,在系统发育树上与维氏气单胞菌聚为一枝。病理组织切片显示肝脏脂肪空泡变性,肾小管变性坏死。药敏实验结果表明CK5、CK9对庆大霉素、氧氟沙星、头孢噻肟、氨曲南、诺氟沙星等药物敏感,对万古霉素、克林霉素、苯唑西林耐药。
Recently, the large number of C. carpio breakout death was suspected to be caused by bacteria after eliminated the parasitic infection. In order to find the pathogenic bacteria which caused the death of C. carpio, the liver, intestine, kidney and other tissues of sick fish were investigated by isolation and purification of bacteria. After gram staining micro physiological, biochemical reaction and molecular biological identification. Two bacteria CK5 and CK9 were separated successfully. Both bacterias were Gram negative bacillus as well as normal growth at 30 ℃. The experiment of lactose, glucose, cellobiose, glutin, ortho - hydroxybenzoic acid were positive. By amplified and sequenced gyrB, aligned sequences with GenBank that results showed the sequence at high homology (99.6% and 99. 0% ) with Aeromonas veronii, phylogenetic tree also indicated with high degree confidcnc. The histopathologic slide display liver axunge vacuolar degeneration and kidney tubules degenerative necrosis. CK5 and CK9 were sensitive to genta, icin, ofloxacin, cefotaxime, aztreonam, norfloxacin, whlie exhibited resistance vancomycin, clindamycin and oxacillin.