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从印度南部患病养殖对虾中分离出的坎贝尔气单胞菌的鉴定及其在卤虫模型中致病性的确立。

Identification of Vibrio campbellii isolated from diseased farm-shrimps from south India and establishment of its pathogenic potential in an Artemia model.

机构信息

Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Osaka, Japan.

Research and Development Centre, Fuso Pharmaceutical Industries Ltd, Osaka, Japan.

出版信息

Microbiology (Reading). 2011 Jan;157(Pt 1):179-188. doi: 10.1099/mic.0.041475-0. Epub 2010 Sep 16.

DOI:10.1099/mic.0.041475-0
PMID:20847009
Abstract

Shrimp diseases are frequently reported to be caused by closely related vibrios, and in many cases they are tentatively but inaccurately identified as Vibrio harveyi and related vibrios. In the present study, 28 biochemically identified V. harveyi-related strains isolated from diseased shrimps were randomly selected for further characterization by molecular tools. Twenty-six strains were identified as Vibrio campbellii and two as V. harveyi by sequence analysis of 16S rRNA and uridylate kinase genes. Haemolysin-gene-based species-specific multiplex PCR also confirmed these results. Experimental challenge studies using Artemia as a model showed that eight isolates were highly pathogenic, three were moderately pathogenic and the remaining 17 were non-pathogenic. Ribotyping with BglI clearly distinguished V. campbellii from V. harveyi, but it failed to separate pathogenic and non-pathogenic clusters. Artemia nauplii challenged with a fluorescently labelled highly pathogenic strain (IPEY54) showed patches in the digestive tract. However, no patches were observed for a non-pathogenic strain (IPEY41). Direct bacterial counts also supported colonization potential for the highly pathogenic strain. To our knowledge, this is the first report on the isolation and accurate identification of large numbers of V. campbellii associated with shrimp disease in aquacultural farms. V. campbellii has long been considered to be non-pathogenic and classified with V. harveyi-related bacteria. However, we show that this species may be an emerging aquaculture pathogen. This study will help to formulate suitable strategies to combat this newly identified pathogen.

摘要

虾病常由密切相关的弧菌引起,在许多情况下,这些弧菌被暂定为哈维氏弧菌和相关弧菌,但鉴定并不准确。在本研究中,从患病虾中随机选择了 28 株经生化鉴定为哈维氏弧菌相关的菌株,并用分子工具进一步进行特征描述。26 株被鉴定为坎贝尔氏弧菌,2 株被鉴定为哈维氏弧菌,这是通过 16S rRNA 和尿嘧啶激酶基因序列分析得出的结果。基于溶血素基因的种特异性多重 PCR 也证实了这些结果。使用卤虫作为模型的实验性攻毒研究表明,有 8 个分离株具有高致病性,3 个具有中等致病性,其余 17 个为非致病性。用 BglI 进行的核糖体分型清楚地区分了坎贝尔氏弧菌和哈维氏弧菌,但它未能将致病性和非致病性聚类分开。用荧光标记的高致病性菌株(IPEY54)攻毒卤虫后,在消化道中出现了斑块。然而,用非致病性菌株(IPEY41)攻毒时则没有观察到斑块。直接细菌计数也支持高致病性菌株的定植潜力。据我们所知,这是首次报道在水产养殖农场中从虾病中分离和准确鉴定大量与坎贝尔氏弧菌相关的弧菌。坎贝尔氏弧菌长期以来一直被认为是非致病性的,并与哈维氏弧菌相关细菌分类在一起。然而,我们表明该物种可能是一种新兴的水产养殖病原体。本研究将有助于制定合适的策略来对抗这种新发现的病原体。

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