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从台湾养殖罗非鱼(奥利亚罗非鱼)分离出的东方诺卡氏弗朗西斯菌亚种的分子特征及致病性

Molecular characterization and pathogenicity of Francisella noatunensis subsp. orientalis isolated from cultured tilapia (Oreochromis sp.) in Taiwan.

作者信息

Pulpipat Theeraporn, Lin Kuo-Hua, Chen Ying-Hao, Wang Pei-Chi, Chen Shih-Chu

机构信息

Department of Veterinary Medicine, College of Veterinary Medicine, National Pingtung University of Science and Technology, Pingtung, Taiwan.

Animal Protection Office, Taoyuan City, Taiwan.

出版信息

J Fish Dis. 2019 May;42(5):643-655. doi: 10.1111/jfd.12964. Epub 2019 Feb 4.

Abstract

Francisella noatunensis subsp. orientalis is a causative agent of systemic granulomatous disease in tilapia. The present study was designed to understand the genetic and phenotypic diversities among Taiwanese Fno isolates obtained from tilapia (n = 17) and green Texas cichlid (Herichthys cyanoguttatus) (n = 1). The enzymatic profiles of the isolates were studied using the API ZYM system. Phylogenetic tree analysis of the 16S rRNA and housekeeping gene and pulsed-field gel electrophoresis (PFGE) were carried out to determine the genotypic characters of all isolates. The phylogenetic tree showed similarity of 99%-100% nucleotide sequences of 16S rRNA and housekeeping genes compared to the Fno references genes from GenBank database. Comparatively, the results revealed an identical profile of enzymatic and PFGE pattern which was distincted from that of F. philomiragia. To understand the pathogenicity, the isolates were intraperitoneal injected to tilapia the gross lesions were observed concomitant with natural outbreak. Median lethal dose upon Nile tilapia and red tilapia were 9.06 × 10 CFU/fish and 2.08 × 10 CFU/fish, respectively. Thus, our data provide understanding the epidemiology of Taiwanese Fno isolates, and help in development of future control and prevention.

摘要

东方诺氏弗朗西斯菌是罗非鱼系统性肉芽肿病的病原体。本研究旨在了解从罗非鱼(n = 17)和德州豹(Herichthys cyanoguttatus)(n = 1)分离得到的台湾东方诺氏弗朗西斯菌菌株之间的遗传和表型多样性。使用API ZYM系统研究了这些菌株的酶谱。对16S rRNA、管家基因进行系统发育树分析,并进行脉冲场凝胶电泳(PFGE)以确定所有菌株的基因型特征。系统发育树显示,与GenBank数据库中的东方诺氏弗朗西斯菌参考基因相比,16S rRNA和管家基因的核苷酸序列相似度为99%-100%。相比之下,结果显示酶谱和PFGE模式与费氏弗朗西斯菌不同。为了解其致病性,将这些菌株腹腔注射到罗非鱼体内,观察到的大体病变与自然爆发情况一致。尼罗罗非鱼和红罗非鱼的半数致死剂量分别为9.06×10 CFU/鱼和2.08×10 CFU/鱼。因此,我们的数据有助于了解台湾东方诺氏弗朗西斯菌菌株的流行病学情况,并有助于未来制定控制和预防措施。

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