☆ 4.7 Article

Construction of a Vibrio alginolyticus hopPmaJ (hop) mutant and evaluation of its potential as a live attenuated vaccine in orange-spotted grouper (Epinephelus coioides)

FISH & SHELLFISH IMMUNOLOGY (2018)

Journal

FISH & SHELLFISH IMMUNOLOGY

Volume 76, Issue -, Pages 93-100

Publisher

ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD

DOI: 10.1016/j.fsi.2018.02.012

Keywords

Vibrio alginolyticus; T3SS; hopPmaJ; Live attenuated vaccine; Epinephelus coioides

Funding

National Natural Science Foundation of China [31402344]
Natural Science Foundation of Guangdong Province [2017A030313174]
Outstanding Young Backbone Teacher Cultivation Program of Guangdong Ocean University [HDYQ2015005]
Project of Enhancing School with innovation of Guangdong Ocean University [GDOU2015050216]
Basic research free exploration project of Shenzhen science and technology innovation Committee

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Abstract

Vibrio alginolyticus, a bacterial pathogen in fish and humans, expresses a type III secretion system (T3SS) that is critical for pathogen virulence and disease development. However, little is known about the associated effectors (T3SEs) and their physiological role. In this study, the T3SE gene hopPmaJ (hop) was cloned from V. alginolyticus wild-type strain HY9901 and the mutant strain HY9901 Delta hop was constructed by the in-frame deletion method. The results showed that the deduced amino acid sequence of V. alginolyticus HopPmaJ shared 78-98% homology with other Vibrio spp. In addition, the HY9901 Delta hop mutant showed an attenuated swarming phenotype and a 2600-fold decrease in the virulence to grouper. However, the HY9901 Delta hop mutant showed no difference in morphology, growth, biofilm formation and ECPase activity. Finally, grouper vaccinated via intraperitoneal (IP) injection with HY9901 Delta hop induced a high antibody titer with a relative percent survival (RPS) value of 84% after challenging with the wild-type HY9901. Real-time PCR assays showed that vaccination with HY9901 Delta hop enhanced the expression of immune-related genes, including MHC-I alpha, MHC-II alpha, IgM, and IL-1 beta after vaccination, indicating that it is able to induce humoral and cell-mediated immune response in grouper. These results demonstrate that the HY9901 Delta hop mutant could be used as an effective live vaccine to combat V. alginolyticus in grouper.

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A live attenuated strain of HY9901AvscB provides protection against Vibrio alginolyticus in pearl gentian grouper (9Epinephelus fuscoguttatus x dEpinephelus lanceolatu)

Huanying Pang, Yunsheng Chang, Hongwei Zheng, Huiming Tan, Shihui Zhou, Fuyuan Zeng, Rowena Hoare, Sean J. Monaghan, Na Wang, Yu Ding

Summary: Vibrio alginolyticus, an important pathogen in fish and humans, expresses a type III secretion system (T3SS) crucial for virulence. A chaperone protein gene vscB was cloned and a mutant strain HY9901ΔvscB was constructed, exhibiting decreased virulence to fish. Transcriptome sequencing identified 95 differentially expressed genes, with 57 up-regulated and 38 down-regulated in the ΔvscB strain. Quantitative RT-PCR analysis showed altered expression of T3SS-related genes in the ΔvscB strain. Vaccination with HY9901ΔvscB induced immune responses in pearl gentian grouper, suggesting it as a potential attenuated vaccine against V. alginolyticus infection.

AQUACULTURE (2022)