Jinyuan Che , Shaojie Hu , Qitong Fang , Binghong Liu , Zhuochen Liu , Cunjie Hu , Lei Wang , Lekang Li , Baolong Bao
{"title":"副溶血性弧菌中不同溶血素基因缺失菌株(ΔhlyA、ΔhlyIII)的构建和特征描述及其毒力评估。","authors":"Jinyuan Che , Shaojie Hu , Qitong Fang , Binghong Liu , Zhuochen Liu , Cunjie Hu , Lei Wang , Lekang Li , Baolong Bao","doi":"10.1016/j.jip.2024.108210","DOIUrl":null,"url":null,"abstract":"<div><div><em>Vibrio parahaemolyticus</em>, a halophilic food-borne pathogen, possesses an arsenal of virulence factors. The pathogenicity of <em>V. parahaemolyticus</em> results from a combination of various virulence factors. <em>HlyA</em> and <em>hlyIII</em> genes are presumed to function in hemolysis, in addition to <em>tdh</em> and <em>trh</em> in <em>V. parahaemolyticus</em>. To confirm the hemolytic function of genes <em>hlyA</em> and <em>hlyIII</em>, Δ<em>hlyA</em> and Δ<em>hlyIII</em> strains of <em>V. parahaemolyticus</em> were separately constructed via homologous recombination. The cytotoxicity and pathogenicity of the Δ<em>hlyA</em> and Δ<em>hlyIII</em> strains were evaluated using a Tetrahymena-<em>Vibrio</em> co-culture model and an immersion challenge in <em>Litopenaeus vannamei</em>. Results indicated that the hemolytic activity of the Δ<em>hlyA</em> and Δ<em>hlyIII</em> strains decreased by approximately 31.4 % and 24.9 % respectively, compared to the WT strain. Both Δ<em>hlyA</em> and Δ<em>hlyIII</em> exhibited reduced cytotoxicity towards Tetrahymena<em>.</em> Then shrimp infection experiments showed LD<sub>50</sub> values for Δ<em>hlyA</em> and Δ<em>hlyIII</em> of 3.06 × 10<sup>8</sup> CFU/mL and 1.23 × 10<sup>8</sup> CFU/mL, respectively, both higher than the WT strain’s value of 2.57 × 10<sup>7</sup> CFU/mL. Histopathological observations revealed that hepatopancreas from shrimps challenged with Δ<em>hlyA</em> and Δ<em>hlyIII</em> exhibited mild symptoms, whereas those challenged with the WT strain displayed severe AHPND. These findings indicate that the Δ<em>hlyA</em> and Δ<em>hlyIII</em> strains are significantly less virulent than the WT strain. In conclusion, both <em>hlyA</em> and <em>hlyIII</em> are vital virulence genes involved in hemolytic and cytotoxic of <em>V. parahaemolyticus</em>.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":null,"pages":null},"PeriodicalIF":3.6000,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Construction and characterization of different hemolysin gene deletion strains in Vibrio parahaemolyticus (ΔhlyA, ΔhlyIII) and evaluation of their virulence\",\"authors\":\"Jinyuan Che , Shaojie Hu , Qitong Fang , Binghong Liu , Zhuochen Liu , Cunjie Hu , Lei Wang , Lekang Li , Baolong Bao\",\"doi\":\"10.1016/j.jip.2024.108210\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><div><em>Vibrio parahaemolyticus</em>, a halophilic food-borne pathogen, possesses an arsenal of virulence factors. The pathogenicity of <em>V. parahaemolyticus</em> results from a combination of various virulence factors. <em>HlyA</em> and <em>hlyIII</em> genes are presumed to function in hemolysis, in addition to <em>tdh</em> and <em>trh</em> in <em>V. parahaemolyticus</em>. To confirm the hemolytic function of genes <em>hlyA</em> and <em>hlyIII</em>, Δ<em>hlyA</em> and Δ<em>hlyIII</em> strains of <em>V. parahaemolyticus</em> were separately constructed via homologous recombination. The cytotoxicity and pathogenicity of the Δ<em>hlyA</em> and Δ<em>hlyIII</em> strains were evaluated using a Tetrahymena-<em>Vibrio</em> co-culture model and an immersion challenge in <em>Litopenaeus vannamei</em>. Results indicated that the hemolytic activity of the Δ<em>hlyA</em> and Δ<em>hlyIII</em> strains decreased by approximately 31.4 % and 24.9 % respectively, compared to the WT strain. Both Δ<em>hlyA</em> and Δ<em>hlyIII</em> exhibited reduced cytotoxicity towards Tetrahymena<em>.</em> Then shrimp infection experiments showed LD<sub>50</sub> values for Δ<em>hlyA</em> and Δ<em>hlyIII</em> of 3.06 × 10<sup>8</sup> CFU/mL and 1.23 × 10<sup>8</sup> CFU/mL, respectively, both higher than the WT strain’s value of 2.57 × 10<sup>7</sup> CFU/mL. Histopathological observations revealed that hepatopancreas from shrimps challenged with Δ<em>hlyA</em> and Δ<em>hlyIII</em> exhibited mild symptoms, whereas those challenged with the WT strain displayed severe AHPND. These findings indicate that the Δ<em>hlyA</em> and Δ<em>hlyIII</em> strains are significantly less virulent than the WT strain. In conclusion, both <em>hlyA</em> and <em>hlyIII</em> are vital virulence genes involved in hemolytic and cytotoxic of <em>V. parahaemolyticus</em>.</div></div>\",\"PeriodicalId\":16296,\"journal\":{\"name\":\"Journal of invertebrate pathology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":3.6000,\"publicationDate\":\"2024-09-27\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of invertebrate pathology\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0022201124001538\",\"RegionNum\":3,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"ZOOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of invertebrate pathology","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0022201124001538","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"ZOOLOGY","Score":null,"Total":0}
Construction and characterization of different hemolysin gene deletion strains in Vibrio parahaemolyticus (ΔhlyA, ΔhlyIII) and evaluation of their virulence
Vibrio parahaemolyticus, a halophilic food-borne pathogen, possesses an arsenal of virulence factors. The pathogenicity of V. parahaemolyticus results from a combination of various virulence factors. HlyA and hlyIII genes are presumed to function in hemolysis, in addition to tdh and trh in V. parahaemolyticus. To confirm the hemolytic function of genes hlyA and hlyIII, ΔhlyA and ΔhlyIII strains of V. parahaemolyticus were separately constructed via homologous recombination. The cytotoxicity and pathogenicity of the ΔhlyA and ΔhlyIII strains were evaluated using a Tetrahymena-Vibrio co-culture model and an immersion challenge in Litopenaeus vannamei. Results indicated that the hemolytic activity of the ΔhlyA and ΔhlyIII strains decreased by approximately 31.4 % and 24.9 % respectively, compared to the WT strain. Both ΔhlyA and ΔhlyIII exhibited reduced cytotoxicity towards Tetrahymena. Then shrimp infection experiments showed LD50 values for ΔhlyA and ΔhlyIII of 3.06 × 108 CFU/mL and 1.23 × 108 CFU/mL, respectively, both higher than the WT strain’s value of 2.57 × 107 CFU/mL. Histopathological observations revealed that hepatopancreas from shrimps challenged with ΔhlyA and ΔhlyIII exhibited mild symptoms, whereas those challenged with the WT strain displayed severe AHPND. These findings indicate that the ΔhlyA and ΔhlyIII strains are significantly less virulent than the WT strain. In conclusion, both hlyA and hlyIII are vital virulence genes involved in hemolytic and cytotoxic of V. parahaemolyticus.
期刊介绍:
The Journal of Invertebrate Pathology presents original research articles and notes on the induction and pathogenesis of diseases of invertebrates, including the suppression of diseases in beneficial species, and the use of diseases in controlling undesirable species. In addition, the journal publishes the results of physiological, morphological, genetic, immunological and ecological studies as related to the etiologic agents of diseases of invertebrates.
The Journal of Invertebrate Pathology is the adopted journal of the Society for Invertebrate Pathology, and is available to SIP members at a special reduced price.