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ORIGINAL RESEARCH article

Front. Microbiol.
Sec. Infectious Agents and Disease
Volume 15 - 2024 | doi: 10.3389/fmicb.2024.1437408
This article is part of the Research Topic Expanded genus Brucella: from taxonomy to clinical manifestations and diagnosis challenges View all 4 articles

Combinatio n of in silico and molecul ar techniques for discr i mination and virulence characterization of marine Brucella ce t i and Brucella pinnipedialis

Provisionally accepted
Guillaume GIRAULT Guillaume GIRAULT 1Luca Freddi Luca Freddi 1Jay Maryne Jay Maryne 1Ludivine Perrot Ludivine Perrot 1Alexandre Dremeau Alexandre Dremeau 1Antoine DRAPEAU Antoine DRAPEAU 1Sabine Delannoy Sabine Delannoy 2Patrick FACH Patrick FACH 2Acacia Ferreira Vicente Acacia Ferreira Vicente 1Virginie MICK Virginie MICK 1Claire PONSART Claire PONSART 1Vitomir Djokic Vitomir Djokic 1*
  • 1 Animla Health Laboratory, French Agency for Food, Environmental and Occupational Health & Safety (ANSES), Maisons-Alfort, France
  • 2 Genomics Platform IdentyPath, Laboratory for Food Safety, ANSES, 94700 Maisons-Alfort, France, Maisons-Alfort, France

The final, formatted version of the article will be published soon.

    Mammals are 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. We compared the DNA and genome sequences of 12 strains isolated from nine marine mammals, with highly zoonotic B. melitensis, B. abortus, B. suis and publicly available genomes of B. ceti and B. pinnipedialis. In silico pipelines were used to detect the antimicrobial resistances (AMR), plasmid and virulence genes (VG) by screening six open-source and one home-made libraries. Our results show that easier-to-use HRM-PCR, Bruce-ladder and Suis-ladder, can separate marine Brucella sp. and results are fully concordant with other molecular methods, such as WGS. However, 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 3 clades according to their preferred host, which was confirmed in WGS. In silico analysis didn’t find any AMR and plasmid genes, suggesting antimicrobial susceptibility of marine Brucella, while presence of VGs btpA gene was variable dependent of clade. The 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.

    Keywords: Bru cella, molecula r biology, Ma rin e mammals, wildlife, molecula r typing

    Received: 23 May 2024; Accepted: 23 Aug 2024.

    Copyright: © 2024 GIRAULT, Freddi, Maryne, Perrot, Dremeau, DRAPEAU, Delannoy, FACH, Ferreira Vicente, MICK, PONSART and Djokic. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: Vitomir Djokic, Animla Health Laboratory, French Agency for Food, Environmental and Occupational Health & Safety (ANSES), Maisons-Alfort, France

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