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A T3SS Regulator Mutant of Vibrio alginolyticus Affects Antibiotic Susceptibilities and Provides Significant Protection to Danio rerio as a Live Attenuated Vaccine
Zhou, Shihui1,2,3,4,5,6; Tu, Xueting1,2,3,4,5,6; Pang, Huanying1,2,3,4,5,6; Hoare, Rowena7; Monaghan, Sean J.7; Luo, Jiajun2,3,4; Jian, Jichan2,3,4,5,6
2020-04-28
Source PublicationFRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY
ISSN2235-2988
Volume10Pages:13
Corresponding AuthorPang, Huanying(phying1218@163.com)
AbstractVibrio alginolyticus is a major cause of Vibriosis in farmed marine aquatic animals and has caused large economic losses to the Asian aquaculture industry in recent years. Therefore, it is necessary to control V. alginolyticus effectively. The virulence mechanism of V. alginolyticus, the Type III secretion system (T3SS), is closely related to its pathogenicity. In this study, the T3SS gene tyeA was cloned from V. alginolyticus wild-type strain HY9901 and the results showed that the deduced amino acid sequence of V. alginolyticus tyeA shared 75-83% homology with other Vibrio spp. The mutant strain HY9901 Delta tyeA was constructed by Overlap-PCR and homologous recombination techniques. The HY9901 Delta tyeA mutant exhibited an attenuated swarming phenotype and an similar to 40-fold reduction in virulence to zebrafish. However, the HY9901 Delta tyeA mutant showed no difference in growth, biofilm formation and ECPase activity. Antibiotic susceptibility test was observed that wild and mutant strains were extremely susceptible to Amikacin, Minocycline, Gentamicin, Cefperazone; and resistant to oxacillin, clindamycin, ceftazidime. In contrast wild strains are sensitive to tetracycline, chloramphenicol, kanamycin, doxycycline, while mutant strains are resistant to them. qRT-PCR was employed to analyze the transcription levels of T3SS-related genes, the results showed that compared with HY9901 wild type, Delta tyeA had increased expression of vscL, vscK, vscO, vopS, vopN, vscN, and hop. Following vaccination with the mutant strain, zebrafish had significantly higher survival than controls following infection with the wild-type HY9901 (71.2% relative percent survival; RPS). Analysis of immune gene expression by qPCR showed that vaccination with HY9901 Delta tyeA increased the expression of IgM, IL-1 beta, IL-6, and TNF-alpha in zebrafish. This study provides evidence of protective efficacy of a live attenuated vaccine targeting the T3SS of V. alginolyticus which may be facilitated by up-regulated pro-inflammatory and immunoglobulin-related genes.
KeywordVibrio alginolyticus live attenuated vaccine type III secretory system tyeA regulator
DOI10.3389/fcimb.2020.00183
Indexed BySCI
Language英语
Funding ProjectShenzhen Science and Technology Project[JCYJ20170818111629778] ; National Key Research and Development Program of China[2018YFD0900501] ; National Natural Science Foundation of China[31402344] ; National Natural Science Foundation of China[31670129] ; Natural Science Foundation of Guangdong Province[2017A030313174] ; Independent Project of SouthernMarine Science and Engineering Guangdong Laboratory (Zhanjiang)[ZJW-2019-05] ; special funds for science technology innovation and industrial development of Shenzhen Dapeng New District[PT202001-13]
WOS Research AreaImmunology ; Microbiology
WOS SubjectImmunology ; Microbiology
WOS IDWOS:000533355300001
PublisherFRONTIERS MEDIA SA
Citation statistics
Document Type期刊论文
Identifierhttp://ir.qdio.ac.cn/handle/337002/167479
Collection实验海洋生物学重点实验室
Corresponding AuthorPang, Huanying
Affiliation1.Guangdong Ocean Univ, Shenzhen Inst, Shenzhen, Peoples R China
2.Guangdong Ocean Univ, Fisheries Coll, Zhanjiang, Peoples R China
3.Guangdong Prov Key Lab Pathogen Biol & Epidemiol, Zhanjiang, Peoples R China
4.Guangdong Key Lab Control Dis Aquat Econ Anim, Zhanjiang, Peoples R China
5.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao, Peoples R China
6.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao, Peoples R China
7.Univ Stirling, Inst Aquaculture, Stirling, Scotland
First Author AffilicationInstitute of Oceanology, Chinese Academy of Sciences
Corresponding Author AffilicationInstitute of Oceanology, Chinese Academy of Sciences
Recommended Citation
GB/T 7714
Zhou, Shihui,Tu, Xueting,Pang, Huanying,et al. A T3SS Regulator Mutant of Vibrio alginolyticus Affects Antibiotic Susceptibilities and Provides Significant Protection to Danio rerio as a Live Attenuated Vaccine[J]. FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY,2020,10:13.
APA Zhou, Shihui.,Tu, Xueting.,Pang, Huanying.,Hoare, Rowena.,Monaghan, Sean J..,...&Jian, Jichan.(2020).A T3SS Regulator Mutant of Vibrio alginolyticus Affects Antibiotic Susceptibilities and Provides Significant Protection to Danio rerio as a Live Attenuated Vaccine.FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY,10,13.
MLA Zhou, Shihui,et al."A T3SS Regulator Mutant of Vibrio alginolyticus Affects Antibiotic Susceptibilities and Provides Significant Protection to Danio rerio as a Live Attenuated Vaccine".FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY 10(2020):13.
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