A MALDI-TOF MS database for fast identification of Vibrio spp. potentially pathogenic to marine mollusks

Type Article
Acceptance Date 2021 IN PRESS
Language English
Author(s) Moussa M.1, Cauvin E.2, Le Piouffle A.3, Lucas O.4, Bidault Adeline5, Paillard Christine5, Benoit F.2, Thuillier B.3, Treilles M.4, Travers Marie-Agnes1, 6, Garcia Celine1
Affiliation(s) 1 : Ifremer, SG2M-LGPMM, Laboratoire de Génétique et Pathologie des Mollusques Marins, F-17390, La Tremblade, France
2 : Labeo-Manche, 1352 avenue de Paris, 50000, Saint-Lô, France
3 : Labocea, Avenue de la Plage des Gueux, 29330, Quimper, France
4 : Qualyse, ZI Montplaisir, 79220, Champdeniers Saint-Denis, France
5 : Univ Brest, CNRS, IRD, Ifremer, UMR6539 LEMAR, F-29280, Plouzané, France
6 : IHPE, Univ. Montpellier, CNRS, Ifremer, Univ. Perpignan Via Domitia, F-34090, Montpellier, France
Source Applied Microbiology and Biotechnology (0175-7598) (Springer Science and Business Media LLC) In Press
DOI 10.1007/s00253-021-11141-0
Keyword(s) Vibrio, MALDI-TOF MS, Database, Marine mollusk pathogens
Abstract

In mollusk aquaculture, a large number of Vibrio species are considered major pathogens. Conventional methods based on DNA amplification and sequencing used to accurately identify Vibrio species are unsuitable for monitoring programs because they are time-consuming and expensive. The aim of this study was, therefore, to develop the MALDI-TOF MS method in order to establish a rapid identification technique for a large panel of Vibrio species. We created the EnviBase containing 120 main spectra projections (MSP) of the Vibrio species that are potentially responsible for mollusk diseases, comprising 25 species: V. aestuarianus, V. cortegadensis, V. tapetis and species belonging to the Coralliilyticus, Harveyi, Mediterranei, and Orientalis clades. Each MSP was constructed by the merger of raw spectra obtained from three different media and generated by three collaborating laboratories to increase the diversity of the conditions and thus obtain a good technique robustness. Perfect discrimination was obtained with all of the MSP created for the Vibrio species and even for very closely related species as V. europaeus and V. bivalvicida. The new EnviBase library was validated through a blind test on 100 Vibrio strains performed by our three collaborators who used the direct transfer and protein extraction methods. The majority of the Vibrio strains were successfully identified with the newly created EnviBase by the three laboratories for both protocol methods. This study documents the first development of a freely accessible database exclusively devoted to Vibrio found in marine environments, taking into account the high diversity of this genus.

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Moussa M., Cauvin E., Le Piouffle A., Lucas O., Bidault Adeline, Paillard Christine, Benoit F., Thuillier B., Treilles M., Travers Marie-Agnes, Garcia Celine A MALDI-TOF MS database for fast identification of Vibrio spp. potentially pathogenic to marine mollusks. Applied Microbiology and Biotechnology IN PRESS. Publisher's official version : https://doi.org/10.1007/s00253-021-11141-0 , Open Access version : https://archimer.ifremer.fr/doc/00680/79224/