Delineating the key virulence factors and intraspecies divergence of Vibrio harveyi via whole-genome sequencing
Publication: Canadian Journal of Microbiology • 17 September 2020 • https://doi.org/10.1139/cjm-2020-0079
Abstract
Vibrio harveyi is one of the major pathogens in aquaculture. To identify the key virulence factors affecting pathogenesis of V. harveyi towards fish, we conducted a field investigation for three representative fish farms infected with V. harveyi. Multilocus sequence typing (MLST) and whole-genome sequencing were conducted to delineate the phylogenetic relationship and genetic divergence of V. harveyi. A total of 25 V. harveyi strains were isolated from the diseased fish and groundwater and were subtyped into 12 sequence types by MLST. Five virulence genes, mshB, pilA, hutR, ureB, and ureG, were variably presented in the sequenced strains. The virulence gene profiles strongly correlated with the distinct pathogenicity of V. harveyi strains, with a strain harboring all five genes exhibiting the highest virulence towards fish. Phenotype assay confirmed that reduced virulence correlated with decreased motility and biofilm formation ability. Additionally, three types of type VI secretion system, namely T6SS1, T6SS2, and T6SS3, were identified in V. harveyi strains, which can be classified into six, four, and 12 subtypes, respectively. In conclusion, the results indicated that the virulence level of V. harveyi is mainly determined by the above virulence genes, which may play vital roles in environmental adaptation for V. harveyi.
Résumé
Vibrio harveyi est l’un des principaux agents pathogènes en aquaculture. Afin d’identifier les principaux facteurs de virulence affectant la pathogenèse de V. harveyi envers les poissons, une enquête de terrain a été réalisée dans trois fermes aquacoles représentatives infectées par V. harveyi. Le typage de séquences multilocus (MLST) et le séquençage du génome entier ont été effectués pour définir la relation phylogénétique et la divergence génétique de V. harveyi. Au total, 25 souches de V harveyi ont été isolées des poissons malades et des eaux souterraines, qui ont pu être sous-typées en 12 types de séquences par MLST. Cinq gènes de virulence, dont mshB, pilA, hutR, ureB et ureG, étaient présents de manière variable dans les souches séquencées. Les profils des gènes de virulence étaient fortement corrélés avec la pathogénicité distincte des souches de V. harveyi, dans lesquelles une souche abritant les cinq gènes présentait la plus forte virulence envers les poissons. L’analyse du phénotype a confirmé qu’une virulence réduite était corrélée à une diminution de la motilité et de la capacité de formation d’un biofilm. En outre, trois types du système de sécrétion de type VI à savoir T6SS1, T6SS2 et T6SS3 ont été identifiés dans les souches V. harveyi, qui peuvent être classées en six, quatre et 12 sous-types, respectivement. En conclusion, les résultats indiquent que le niveau de virulence de V. harveyi est principalement déterminé par les gènes de virulence susmentionnés, qui peuvent jouer un rôle vital dans l’adaptation environnementale de V. harveyi.
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Canadian Journal of Microbiology
Volume 67 • Number 3 • March 2021
Pages: 231 - 248
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Received: 15 February 2020
Revision received: 21 April 2020
Accepted: 21 May 2020
Published online: 17 September 2020
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Songzhe Fu, Ping Ni, Qian Yang, Huizhi Hu, Qingyao Wang, Shigen Ye, and Ying Liu. Delineating the key virulence factors and intraspecies divergence of Vibrio harveyi via whole-genome sequencing. Canadian Journal of Microbiology.
67(3): 231-248. https://doi.org/10.1139/cjm-2020-0079
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