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Host gut-derived Bacillus probiotics supplementation improves growth performance, serum and liver immunity, gut health, and resistive capacity against vibrio harveyi infection in hybrid grouper(♀Epinephelus fuscoguttatus × ♂Epinephelus lanceolatus)

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Animal Nutrition 2023年第3期14卷 163-184页

作者： Kwaku Amoah Beiping Tan Shuang Zhang Shuyan Chi Qihui Yang Hongyu Liu Yuanzhi Yang Haitao Zhang Xiaohui Dong Laboratory of Aquatic Animal Nutrition and Feed College of FisheriesGuangdong Ocean UniversityZhanjiangGuangdong 524088China Aquatic Animals Precision Nutrition and High-Efficiency Feed Engineering Research Centre of Guangdong Province ZhanjiangGuangdong 524088China Key Laboratory of Aquatic Livestock and Poultry Feed Science and Technology in South ChinaMinistry of AgricultureZhanjiangGuangdong 524000China

Several reports have revealed the vital role that probiotics play in fish growth and ***,few works are available for host gut-derived probiotics on the growth,immunity,and gut microbiota of fish,especially in hybrid grouper (♀Epinephelus fuscoguttatus×♂Epinephelus lanceolatus) due to their isolation difficulty and functional *** study aimed at assessing 3 host gut-derived Bacillus species?effects on the growth,immune and antioxidant-biochemical responses,haematological parameters,intestinal morphology,immune-related gene expression,gut microbiota,and disease resistance against vibrio harveyi in hybrid grouper.A total of 480 hybrid grouper (initial weight=9.03±0.02 g) were randomly allotted into 4 groups,namely,the group fed a basal diet without probiotic inclusion (control,B0),the group fed the basal diet with Bacillus velezensis GPSAK4 (BV),the group fed the basal diet with Bacillus subtilis GPSAK9 (BS),and the group fed the basal diet with Bacillus tequilensis GPSAK2 (BT) strains at 1.0×10^(9)CFU/*** a 6-week feeding trial,the results revealed significant improvements (P<0.05) in the growth performance,whole fish-body proximate composition,blood haematological parameters,serum,liver,and intestinal biochemical indexes,intestinal morphology,and protection against *** pathogen in the probiotic-treated groups compared with the ***,the expressions of intestinal tight junction genes (occludin and ZO1),pro-and anti-inflammatory genes,including IL1β,IL6,IL8,TNFa,MyD88,IL10,and TGFβ,were upregulated (P<0.05) after Bacillus species *** gut-derived Bacillus supplementation shaped the gut microbiota by significantly increasing (P<0.05) the relative abundance of Proteobacteria,Bacteroidetes,Actinobacteria (except the BS group),Acidobacteria(except the BT group),Cyanobacteria (except the BV and BT groups),and Verrucomicrobia phyla,as well as known beneficial genera (Romboutsia,Turicibacter,Epulopiscium,Clostr

关键词： Hybrid grouper(♀Epinephelus fuscoguttatus×♂E.lanceolatus) Host-associated probiotics Immune and antioxidant-biochemical responses Gut microbiota Immune-related gene expression vibrio harveyi

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Characterization of Caspase Gene Family Members in Spotted Sea Bass(Lateolabrax maculatus)and Their Expression Profiles in Response to vibrio harveyi Infection

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中国海洋大学学报:英文版 2023年第5期22卷 1370-1382页

作者： YANGLANG Arat WEN Haishen MAO Xuebin TIAN Yuan WANG Lingyu LI Jinku QI Xin SRISAPOOME Prapansak LI Jifang LI Yun The Key Laboratory of Mariculture(Ocean University of China) Ministry of EducationQingdao 266003China Laboratory of Aquatic Animal Health Management Department of AquacultureFaculty of FisheriesKasetsart UniversityBangkok 10900Thailand

The caspase gene family is a crucial gene cluster that regulates apoptosis which contribute to programmed cell death,cell proliferation and differentiation,and several immune *** our study,a complete set of 12 caspase genes were identified in spotted sea bass Lateolabrax *** genes were divided into three subfamilies:2 inflammatory caspases(casp-1 and casp-14-like),5 apoptosis initiators(casp-2,casp-8a,casp-8b,casp-9,and casp-10),and 5 apoptosis executioners(casp-3a,casp-3b,casp-3-like,casp-6,and casp-7).Their phylogenetic relationships,synteny and gene structures were systematically ***,the relative expression profiles of the caspase family members in the liver,intestine,head kidney,and spleen were measured by q PCR after infection with vibrio *** results showed that the overall mRNA levels of the caspase genes were dramatically increased after *** infection,and the expression patterns varied among genes and *** caspase genes underwent pronounced expression changes in the head kidney and spleen than in the liver or intestine,mainly after 48 h of the ***,casp-3a,casp-3b,casp-3-like,casp-6,casp-7,casp-8a,casp-8b,casp-10,and casp-14-like in the head kidney,and casp-3-like,casp-6,casp-7,and casp-14-like in the spleen,were the most responsive caspase genes which may contribute significantly to immune regulation in spotted sea ***,the apoptosis level in head kidney and spleen after infection were examined using the Caspase *** study provides a systemic overview of the caspase gene family in spotted sea bass after *** infection and lays a foundation for further deciphering the biological roles of these caspase genes.

关键词： caspase gene family spotted sea bass vibrio harveyi gene expression apoptosis

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Disruption of chemotaxis-related genes affects multiple cellular processes and the virulence of pathogenic vibrio harveyi

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Acta Oceanologica Sinica 2013年第8期32卷 55-60页

作者： QIN Yingxue YAN Qingpi SU Yongquan LI Haiping ZOU Wenzheng Key Laboratory of Healthy Mariculture for the East China Sea Ministry of Agriculture Jimei University Fisheries College Jimei University College of Ocean and Earth Science Xiamen University

Chemotactic motility is involved in the virulence of many pathogenic bacteria. In order to understand the role of chemotactic motility of vibrio harveyi in cellular processes and virulence, mini-TnlO/Kan transposon-induced mutants with deficient chemotactic motility were constructed, screened, and iden- tified. Sequence analysis revealed that the 465-bp fragment （GenBank accession number HM630274） fank- ing the transposon insertion site in mutant TS-CM1 had the highest identity （96.9%） with a hypothetical protein gene of V. harveyiATCC BAA-1116 and the second-highest identity （91.8%） with the pgk gene of V. parahaemolyticus RIMD 2210633. In another mutant, TS-CM2, 356 bp of transposon-flanking sequence （GenBank accession number HM630275） also showed the highest identity （94.6%） with a hypothetical pro- tein gene of V. harveyi ATCC BAA-1116 and the second-highest identity （92.4%） with the flaB gene of V. alginolyticus HY9901. Studies on virulence-related biological characteristics such as growth, motility, adhe- sion, and infectivity of the mutants showed that disruption of either the flagellin gene or energy metabolism gene led to subsequent loss of chemotactic motility and changes in growth, motility, adhesion, and viru- lence of the pathogenic E harueyi. Hence, the flagellin gene and crucial energy metabolism gene played an important role in the chemotactic motility of V. harveyi.

关键词： chemotactic motility vibrio harveyi transposon virulence

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Transcriptional responses to starvation of pathogenic vibrio harveyi strain DY1

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Journal of Oceanology and Limnology 2020年第2期38卷 579-587页

作者： LIU Xiaodan GAO Xiaojian CHEN Nan ZHANG Yingying LI Xixi ZHANG Yue ZHANG Xiaojun College of Animal Science and Technology Yangzhou University

vibrio harveyi is a pathogen of various aquatic organisms that has been recently associated with massive mortality episodes in the aquaculture *** outbreaks of vibriosis are closely correlated with the capacity of this bacterial species to survive long-term starvation *** study the regulation mechanism of gene expression at the transcriptional level in *** under starvation conditions,the transcriptomic response profiles were determined of the Portunus trituberculatus pathogen *** strain DY1 under normal conditions and after four weeks of starvation.A total of 4679 and 4661 genes were expressed in the non-starved and starved cells,*** significantly differentially expressed genes(DEGs)between non-starved and starved groups were identified,in which 255 genes were up-regulated and 411 genes were *** analysis and KEGG enrichment analysis were used to analyze the DEGs and revealed the involvement of these DEGs in many pathways,including ABC transporters,flagellum assembly,and fatty acid *** DEGs were randomly selected and their expression levels were confirmed by quantitative real-time PCR(qRT-PCR).This is the first comprehensive transcriptomic analysis of starvation ef fects in *** findings will facilitate future study on stress adaptation and survival mechanisms of ***.

关键词： vibrio harveyi starvation stress transcriptome sequencing differentially expressed genes adaptation and survival mechanisms

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Antimicrobial peptide hepcidin contributes to host defense of Centropristis striata against vibrio harveyi challenge

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Acta Oceanologica Sinica 2021年第6期40卷 61-66页

作者： Xiaowan Ma Ying Qiao Yanxiang Shao Chao Chen Key Laboratory of Tropical Marine Ecosystem and Bioresource Fourth Institute of OceanographyMinistry of Natural ResourcesBeihai 536000China Yellow Sea Fisheries Research Institute Chinese Academy of Fishery SciencesQingdao 266071China

Hepcidins are small cysteine-rich antimicrobial peptides that play a vital role in immunity against pathogen ***,a hepcidin(Cshep)from Centropristis striata was described,which is considered as a valuable aquaculture marine species in *** open reading frame consisted of 273 *** conserved cysteine residues were *** analysis showed that Cshep had a relatively close relationship with the hepcidin from Epinephelus *** real-time PCR analysis demonstrated that Cshep was highly expressed in liver and significantly up-regulated when challenged with vibrio *** addition,the synthetic Cshep peptide had a high antimicrobial activity against ***,but low against other pathogenic bacteria tested in this *** killing kinetics analysis revealed that Cshep had a fast bactericidal effect on *** results suggested that Cshep may be involved in the immune response of *** against *** infection.

关键词： Centropristis striata hepcidin antimicrobial peptides antimicrobial activity vibrio harveyi

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Antigenicity analysis of vibrio harveyi TS-628 strain

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Frontiers in Biology 2007年第3期2卷 263-267页

作者： QIN Yingxue WANG Jun WANG Shifeng YAN Qingpi Department of Oceanography Institute of Subtropic OceanXiamen UniversityXiamen 361000China Fisheries College Jimei UniversityXiamen 361021China

vibrio harveyi,the major causative agent of vib-riosis,affects a diverse range of marine cultured organisms over a wide geographical ***,reports about screening the effective antigen and research on vaccines of *** are ***,lipopolysaccharide(LPS)and outer membrane proteins(OMP)are major immunogenic antigens in many Gram-negative *** this study,the flagellin,OMP and LPS of the *** TS-628 strain iso-lated from infected groupers were extracted and Western blot analysis was used to detect the antigenicity of these *** of the Western blot assay reveal that there are four positive flagellin bands:35 kDa,38 kDa,43 kDa,and 52 kDa,of which the 43 kDa and 52 kDa bands displayed the strongest positive *** are five positive OMPbands about 35 kDa,38 kDa,43 kDa,47 kDa,and 52 kDa,of which the 43 kDa appeared to have the strongest positive reaction although the other four proteins also displayed strong ***,LPS is Western *** results indicate that the 43 kDa and 52 kDa flagellin and OMP of size 43 kDa,52 kDa can be candidates for developing vaccines against ***.

关键词： vibrio harveyi flagellin LPS OMP antigenicity

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An Extracellular Oligopeptide Permease May Be a Potential Virulence Factor of vibrio harveyi

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Journal of Ocean University of China 2011年第4期10卷 343-350页

作者： HE Qingfang CHEN Jixiang LI Caifeng Department of Marine Biology College of Marine Life SciencesOcean University of ChinaQingdao 266003P.R.China Oil Production Technology Research Institute Shengli Oilfield Company of SINOPECDongying 257000P.R.China

An oligopeptide permease A(OppA)was purified from the extracellular product of vibrio harveyi *** molecular weight of the purified protein was estimated to be 58 kDa on *** purified protein showed phospholipase C activity at the optimal values of temperature 50℃ and pH *** enzymatic activity decreased when the temperature increased to 40℃.The N-terminal sequence of the purified protein was determined as ADVPAGTKLA,which is similar to that of *** OppA pre-cursor gene was cloned from the genome of *** *** gene consisted of 1665 base pairs and encoded a 554 amino acid polypeptide,which showed a high similarity to those of the OppAs of *** and other vibrio *** gene was subcloned into pET-28a(+)and expressed in Escherichia *** expressed recombinant protein was purified by Ni-NTA metal affinity *** expressed recombinant protein showed a 58 kDa band on SDS-PAGE and exhibited phospholipase C activity with the optima of temperature 50℃ and pH *** purified protein was toxic to the flounder gill *** OppA mutant of *** SF-1 was constructed by homologous *** mutant strain was less virulent when it was intraperitoneally inoculated to zebra fish,with the LD50 of 5.46×105 CFU fish-1,compared to 3.11×104 CFU fish-1 of the wild-type strain,which indicated that the OppA might play an important role in the pathogenicity of ***.

关键词： vibrio harveyi oligopeptide permease phospholipase C pathogenicity

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In vitro biofilm forming capacity on abiotic contact surfaces by outbreak-associated vibrio harveyi strains

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Journal of Coastal Life Medicine 2014年第2期2卷 132-140页

作者： Pallaval Veera Bramha Chari Kuchipudi Viswadeepika Bottu Anand Kumar Department of Biotechnology Krishna UniversityMachilipatnam-521001Andhra PradeshIndia

Objective:To evaluate the in vitro biofilm forming capacity on abiotic food contact surfaces by vibrio harveyi(***)***:Thirty six Gram-negative *** strains were isolated from various street vended seafood outlets in a food processing line and evaluated for their ability to produce mucoid biofilms on food contact surfaces using a microplate *** characterization of mucoid biofilm producing *** strains were screened on Congo red agar,thiosulfate-citrate-bile salts-sucrose agar and tryptic soy agar,***:Only five *** strains(14%)were mucoid biofilm producers characterized by formation of black colonies,whereas the remaining 31 strains(86%)were not capable of producing biofilm characterized by formation of red colonies or pinkish-red colonies with darkening at the *** morphological,physiological and biochemical characteristics of these isolates were studied using standard *** identification was confirmed by polymerase chain reaction targeted to species-specific polymerase chain reaction primers VH-1 and VH-2 corresponding to variable regions of *** 16S rRNA *** the biofilm-forming strains showed resistance to at least three antimicrobial compounds *** strains isolated from various seafood were able to form biofilms of different capacity,and the strains VB267,VB238 and VB166 isolated from cat fish,shrimp and eel fish exhibited significantly greater biofilm forming ability compared to other ***:It can be concluded from the present study that the strain VB166 was able to better attach and form subsequent biofilms on glass and stainless steel compared to high density *** properties allow these bacteria to survive,proliferate and persist in street vended seafood outlets.

关键词： Mucoid biofilm Congo red agar vibrio harveyi Polymerase chain reaction High density polyethylene

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In vitro Inhibitory Activity of Shisandra chinensis and Polygonatum sibiricum against vibrio harveyi and Its Biofilms

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Agricultural Biotechnology 2016年第4期5卷 57-60页

作者： Zhifeng HUANG Huanying PANG Yang HUANG Yucong HUANG Yishan LU Jichang JIAN College of Fisheries Guangdong Ocean University/Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals/Key Laboratory of Diseases Controlling for Aquatic Economic Animals of Guangdong Higher Education InstitutionsZhanjiang 524088China

[Objective] This study aimed to analyze the in vitro inhibitory activity of Shisandra chinensis and Polygonatum sibiricum against vibrio harveyi and its biofilms. [Result] By agar diffusion test, in vitro inhibitory activity of 5. chinensis and P. sibiricum against V. harveyi was investigated. The minimal inhibitory concentration （ MIC） and minimal bactericidal concentration （MBC） of 5. chinensis and P. sibiricum against V. harveyi were determined by doubling dilution meth-od. The inhibitory activity of 5. chinensis and P. sibiricum on the formation of V. harveyi biofilms was evaluated by modified MTT assay. [ Result ] Both 5. chinen-sis and P. sibiricum had inhibitory activity against V. harveyi. The inhibition zone diameter of 5. chinensis against V. harveyi was 17. 95 mm; MIC and MBC of 5. chinensis were both 3.125 mg/ml. The inhibition zone diameter of P. sibiricum against V. harveyi was 12. 22 mm; MIC and MBC of P. sibiricum were 3.125 and 6.250 mg/ml, respectively. When the concentration was higher than 6. 25 mg/ml, 5. chinensis decoction had extremely significant inhibitory activity against V. harveyi （P 〈 0. 01） ; when the concentration was higher than 3. 125 mg/ml, P. sibiricum had extremely significant inhibitory activity against V. harveyi （P 〈0. 01）. [ Conclusion] 5. chinensis and P. sibiricum could significantly inhibit V. harveyi and its biofilms.

关键词： Shisandra chinensis Polygonatum sibiricum vibrio harveyi In vitro inhibitory activity Biofilm

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Identification of novel cross immunogenic proteins among vibrio alginolyticus,vibrio harveyi and vibrio parahaemolyticus by immunoproteomic analysis

Identification of novel cross immunogenic proteins among Vib...

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热带海洋科学学术研讨会暨第八届广东海洋湖沼学会、第七届广东海洋学会会员代表大会

作者： Huanying Pang Shuanghu Cai Yucong Huang Jufen Tang Jichang Jian Yishan Lu Zaohe Wu College of Fishery Guangdong Ocean University Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals Guangdong Key Laboratory of Control for Diseases of Aquatic Economic Animals Zhongkai University of Agriculuture and Engineering

In this paper,the two-dimensional electrophoresis(2-DE) gels of vibrio alginolyticus,vibrio harveyi and vibrio parahaemolyticus were probed with Epinephelus coioides anti-V,alginolyticus sera by Western blotting respectively and accordingly 56,58 and 48 immunogenic protein spots were *** matching analysis for the three Western blotting membranes,6 cross immunogenic spots for three vibrio species were *** were Outer membrane protein W(OmpW),dihydrolipoamide dehydrogenase(DLD),succinate dehydrogenase flavoprotein subunit(SDHA),elongation factor Ts(Ts),peptide ABC transporter periplasmic peptide-binding ptotein(ABC) and phosphoenolpyruvate carboxykinase(PEPCK).Three of the proteins,DLD,Ts and ABC were also used to evaluate the cross protective function for *** with a bacterial immunization challenging *** showed the relative percent survival rate against V. alginolyticus,*** and *** was 80%,68%and 68%respectively for each experiment ***, vaccination with Ts and peptide ABC transporter periplasmic peptide-binding ptotein induces low protective immune response in ***,this work may provide potential cross protective vaccine candidate antigens for vibrio species

关键词： vibrio alginolyticus vibrio harveyi vibrio parahaemolyticus cross immunogenic proteins immunoproteomic

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