Vibrio parahaemolyticus becomes lethal to post-larvae shrimp via acquiring novel virulence factors

文献类型： 外文期刊

第一作者： Liu, Shuang

作者： Liu, Shuang;Wang, Wei;Jia, Tianchang;Xin, Lusheng;Xu, Ting-ting;Wang, Chong;Xie, Guosi;Luo, Kun;Kong, Jie;Zhang, Qingli;Liu, Shuang;Wang, Wei;Jia, Tianchang;Xin, Lusheng;Xu, Ting-ting;Wang, Chong;Xie, Guosi;Luo, Kun;Kong, Jie;Zhang, Qingli;Liu, Shuang;Xin, Lusheng;Xu, Ting-ting;Xie, Guosi;Kong, Jie;Zhang, Qingli;Li, Jun

作者机构：

关键词： translucent post-larvae disease (TPD); Vibrio parahaemolyticus-causing TPD (Vp (TPD)); shrimp; novel virulence factor; Vibrio high virulent protein (VHVP)

期刊名称：MICROBIOLOGY SPECTRUM （ 影响因子：3.7； 五年影响因子：5.9 ）

ISSN： 2165-0497

年卷期： 2023 年

页码：

收录情况： SCI

摘要： Translucent post-larvae disease (TPD), caused by Vibrio parahaemolyticus (Vp (TPD)), has become an emerging shrimp disease, affecting more than 70%-80% of coastal shrimp nurseries in China in spring 2020. Here, we investigated the key virulence factors of Vp (TPD) by analyzing protein fragments, related genomic information, as well as experimental challenge tests. After investigating the toxic effects of purified protein fragments with different molecular weights (MWs) from Vp (TPD,) we found that only the protein fragments with MW >100 kDa showed similar lethality to live Vp (TPD) in the experimental challenge test using post-larvae shrimp. Meanwhile, similar histopathological changes exhibiting in the hepatopancreas and midgut of the diseased individuals were observed in the post-larvae shrimp challenged with either bacterial protein fragments (MW >100 kDa) or live Vp (TPD). Based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometry analyses, two novel proteins, Vibrio high virulent protein (VHVP)-1 and VHVP-2, were identified as the candidates of key virulence factors to cause TPD. Moreover, VHVP-1 and VHVP-2 were found to be encoded by two genes (vhvp-1 and vhvp-2) tandemly located on a 187,791-bp plasmid and were predicted to depend on the same promoter following a comparative genomic analysis. Further epidemiological investigation and challenge test indicated that the V. parahaemolyticus isolate carrying only the vhvp-1 gene and lacking vhvp-2 gene could not cause mortality of experimental Penaeus vannamei post-larvae. The mutant (Delta vhvp-2) by deleting vhvp-2 gene could only cause 4.92% of accumulative mortality of post-larvae that is similar to the non-Vp (TPD) Vibrio strain. Additionally, the complemented strains, Delta vhvp-2/pBT3-vhvp-2 and Delta vhvp-2/pwtCas9-vhvp-2, showed similar virulence to the wild-type Vp (TPD). The results demonstrated that V. parahaemolyticus becomes lethal to post-larval shrimp by acquiring the VHVP-2 virulence factor. This study sheds light on further investigations of the pathogenic mechanism of Vp (TPD) and the development of strategies for early diagnosis of TPD in shrimp hatcheries.

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