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.Key points• Development of a MALDI-TOF MS database to quickly affiliate Vibrio species.• Increase of the reactivity when faced with Vibrio associated with mollusk diseases.• Validation of MALDI-TOF MS as routine diagnostic tool.
Highlights
The greatest worldwide threat to the future of marine aquaculture is disease
A large number of bacterial species associated with marine organisms in symbiotic or pathogenic relationships belong to the genus Vibrio (Paillard et al 2004; Thompson et al 2004), which contains more than 100 species distributed across at least twenty clades (Al-Saari et al 2015; Sawabe et al 2013)
For the first time, a MALDI-TOF main spectra (MS) database was built collecting most of the Vibrio species involved in mollusk diseases
Summary
The greatest worldwide threat to the future of marine aquaculture is disease. In particular, there are an increasing number of reports of bacterial infections impacting marine aquaculture production, as well as novel descriptions of new bacterial pathogens. Appl Microbiol Biotechnol (2021) 105:2527–2539 some of these species can affect all of the growth stages (i.e., larval, juvenile, and adult) of their mollusk hosts, whereas others are more specialized (Beaz-Hidalgo et al 2010; Paillard et al 2004). These bacteria are recognized as the most diverse known marine bacterial group due to their presence in various niches in the environment, as well as their metabolic and ecologic versatility. The characterization, classification, and identification of Vibrio species are problematic, especially in very closely related species
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