{"title":"Combination of in silico and molecular techniques for discrimination and virulence characterization of marine Brucella ceti and Brucella pinnipedialis","authors":"Guillaume Girault, Luca Freddi, Maryne Jay, Ludivine Perrot, Alexandre Dremeau, Antoine Drapeau, Sabine Delannoy, Patrick Fach, Acacia Ferreira Vicente, Virginie Mick, Claire Ponsart, Vitomir Djokic","doi":"10.3389/fmicb.2024.1437408","DOIUrl":null,"url":null,"abstract":"IntroductionMammals are the main hosts for <jats:italic>Brucella</jats:italic> sp., agents of worldwide zoonosis. Marine cetaceans and pinnipeds can be infected by <jats:italic>Brucella ceti</jats:italic> and <jats:italic>B. pinnipedialis</jats:italic>, respectively. Besides classical bacteriological typing, molecular approaches such as MLVA, MLSA, and whole-genome sequencing (WGS) can differentiate these species but are cumbersome to perform.MethodsWe compared the DNA and genome sequences of 12 strains isolated from nine marine mammals, with highly zoonotic <jats:italic>B. melitensis</jats:italic>, <jats:italic>B. abortus</jats:italic>, and <jats:italic>B. suis</jats:italic>, and the publicly available genomes of <jats:italic>B. ceti</jats:italic> and <jats:italic>B. pinnipedialis. In silico</jats:italic> pipelines were used to detect the antimicrobial resistance (AMR), plasmid, and virulence genes (VGs) by screening six open-source and one home-made library.Results and discussionOur results show that easier-to-use HRM-PCR, Bruce-ladder, and Suis-ladder can separate marine <jats:italic>Brucella</jats:italic> sp., and the results are fully concordant with other molecular methods, such as WGS. However, the restriction fragment length polymorphism (RFLP) method cannot discriminate between <jats:italic>B. pinnipedialis</jats:italic> and <jats:italic>B. ceti</jats:italic> B1-94-like isolates. MLVA-16 results divided the investigated strains into three clades according to their preferred host, which was confirmed in WGS. <jats:italic>In silico</jats:italic> analysis did not find any AMR and plasmid genes, suggesting antimicrobial susceptibility of marine <jats:italic>Brucella</jats:italic>, while the presence of the VGs <jats:italic>btpA</jats:italic> gene was variable dependent on the clade.ConclusionThe HRM-PCR and Suis-ladder are quick, easy, and cost-effective methods to identify marine <jats:italic>Brucella</jats:italic> sp. Moreover, <jats:italic>in silico</jats:italic> genome analyses can give useful insights into the genetic virulence and pathogenicity potential of marine <jats:italic>Brucella</jats:italic> strains.","PeriodicalId":12466,"journal":{"name":"Frontiers in Microbiology","volume":null,"pages":null},"PeriodicalIF":4.0000,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Frontiers in Microbiology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.3389/fmicb.2024.1437408","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
IntroductionMammals are the main hosts for Brucella sp., agents of worldwide zoonosis. Marine cetaceans and pinnipeds can be infected by Brucella ceti and B. pinnipedialis, respectively. Besides classical bacteriological typing, molecular approaches such as MLVA, MLSA, and whole-genome sequencing (WGS) can differentiate these species but are cumbersome to perform.MethodsWe compared the DNA and genome sequences of 12 strains isolated from nine marine mammals, with highly zoonotic B. melitensis, B. abortus, and B. suis, and the publicly available genomes of B. ceti and B. pinnipedialis. In silico pipelines were used to detect the antimicrobial resistance (AMR), plasmid, and virulence genes (VGs) by screening six open-source and one home-made library.Results and discussionOur results show that easier-to-use HRM-PCR, Bruce-ladder, and Suis-ladder can separate marine Brucella sp., and the results are fully concordant with other molecular methods, such as WGS. However, the restriction fragment length polymorphism (RFLP) method cannot discriminate between B. pinnipedialis and B. ceti B1-94-like isolates. MLVA-16 results divided the investigated strains into three clades according to their preferred host, which was confirmed in WGS. In silico analysis did not find any AMR and plasmid genes, suggesting antimicrobial susceptibility of marine Brucella, while the presence of the VGs btpA gene was variable dependent on the clade.ConclusionThe HRM-PCR and Suis-ladder are quick, easy, and cost-effective methods to identify marine Brucella sp. Moreover, in silico genome analyses can give useful insights into the genetic virulence and pathogenicity potential of marine Brucella strains.
期刊介绍:
Frontiers in Microbiology is a leading journal in its field, publishing rigorously peer-reviewed research across the entire spectrum of microbiology. Field Chief Editor Martin G. Klotz at Washington State University is supported by an outstanding Editorial Board of international researchers. This multidisciplinary open-access journal is at the forefront of disseminating and communicating scientific knowledge and impactful discoveries to researchers, academics, clinicians and the public worldwide.