Tsai Ming-An, Wang Pei-Chyi, Liaw Li-Ling, Yoshida Terutoyo, Chen Shih-Chu
Department of Veterinary Medicine, National Pingtung University of Science and Technology, Pingtung, Taiwan.
Dis Aquat Organ. 2012 Dec 3;102(1):43-51. doi: 10.3354/dao02516.
Seventy-six Taiwanese bacterial isolates including 74 from diseased, cultured, aquatic animals (54 grey mullet Mugil cephalus, 3 basket mullet Chelon alatus, 2 tilapia Oreochromis niloticus, 1 grouper Epinephelus coioides, 2 yellowfin seabream Acanthopagrus latus, 1 Borneo mullet Chelon macrolepis, 1 bullfrog Rana catesbeiana, 1 Japanese eel Anguilla japonica, and 9 giant freshwater prawns Macrobrachium rosenbergii), 1 wild-caught seafood species (squid muscle collected from a restaurant) and 1 human isolate (from a patient with a history of consuming raw squid in the previously mentioned restaurant), all collected between 1999 and 2006, were confirmed by PCR assay to be Lactococcus garvieae. The phenotypic characterization was determined by rabbit anti-KG+ and KG- serums, and 74 of the 76 Taiwanese strains displayed a KG- phenotype. The genetic characterization was investigated by pulsed-field gel electrophoresis (PFGE). Genomic DNA was digested with restriction endonucleases ApaI and SmaI and separated by PFGE. Ten different L. garvieae pulsotypes were identified. Predominant pulsotypes A1a/S1a were obtained from >96% of strains (52 of 54) from grey mullet, demonstrating a clonal dissemination of L. garvieae in grey mullet in Taiwan. In experimental challenges with grey mullet and tilapia, L. garvieae pulsotypes A1/S1 and A11/S11 showed higher virulence compared with other pulsotypes.
76株台湾细菌分离株,其中74株来自患病的养殖水生动物(54条乌头鲻、3条篮鲻、2条尼罗罗非鱼、1条巨石斑鱼、2条黄鳍鲷、1条婆罗洲鲻、1只牛蛙、1条日本鳗鲡和9只罗氏沼虾)、1种野生捕捞的海鲜品种(从一家餐馆采集的鱿鱼肌肉)和1株人类分离株(来自曾在上述餐馆食用生鱿鱼的患者),这些样本均在1999年至2006年间采集,经PCR检测确认为加氏乳球菌。通过兔抗KG +和KG -血清进行表型鉴定,76株台湾菌株中有74株表现出KG -表型。通过脉冲场凝胶电泳(PFGE)进行遗传特征分析。基因组DNA用限制性内切酶ApaI和SmaI消化,然后通过PFGE分离。鉴定出10种不同的加氏乳球菌脉冲型。优势脉冲型A1a/S1a来自超过96%(54条中的52条)的乌头鲻菌株,表明加氏乳球菌在台湾乌头鲻中呈克隆传播。在对乌头鲻和罗非鱼的实验性攻毒中,加氏乳球菌脉冲型A1/S1和A11/S11与其他脉冲型相比显示出更高的毒力。