Abstract
BackgroundMatrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) biotyping is an innovative strategy, applied successfully for the identification of numerous arthropod families including mosquitoes. The effective mosquito identification using this emerging tool was demonstrated possible at different steps of their life-cycle, including eggs, immature and adult stages. Unfortunately, for species identification by MS, the euthanasia of the mosquito specimen is required.MethodsTo avoid mosquito euthanasia, the present study assessed whether aedine mosquitoes could be identified by MALDI-TOF MS biotyping, using their respective exuviae. In this way, exuviae from the fourth-instar and pupal stages of Aedes albopictus and Aedes aegypti were submitted to MALDI-TOF MS analysis.ResultsReproducible and specific MS spectra according to aedine species and stage of exuviae were observed which were objectified by cluster analyses, composite correlation index (CCI) tool and principal components analysis (PCA). The query of our reference MS spectra database (DB) upgraded with MS spectra of exuviae from fourth-instar larvae and pupae of both Aedes species revealed that 100% of the samples were correctly classified at the species and stage levels. Among them, 93.8% (135/144) of the MS profiles reached the threshold log score value (LSV > 1.8) for reliable identification.ConclusionsThe extension of reference MS spectra DB to exuviae from fourth-instar and pupal stages made now possible the identification of mosquitoes throughout their life-cycle at aquatic and aerial stages. The exuviae presenting the advantage to avoid specimen euthanasia, allowing to perform complementary analysis on alive mosquitoes.
Highlights
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) biotyping is an innovative strategy, applied successfully for the identification of numerous arthropod families including mosquitoes
The number of MS peaks detected between the exuviae stages was equivalent between the fourthinstar and pupal stages, with 85 ± 8 and 84 ± 6 MS peaks, respectively, irrespective of the species
To assess the reproducibility and specificity of MS spectra from fourth-instar and pupal exuviae according to species, cluster analysis was performed
Summary
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) biotyping is an innovative strategy, applied successfully for the identification of numerous arthropod families including mosquitoes. Nebbak and Almeras Parasites Vectors (2020) 13:161 and are competent vectors for numerous human and animal pathogens [8] These aedine species invaded several European countries like Switzerland [9], Germany [10], Austria [2], France [11] and Italy [12]. The establishment of these Aedes species in Europe participated to the emergence of autochthonous human cases of dengue and chikungunya [13, 14] To face of this growing vulnerability of European countries to MBDs, the implementation of guidelines for surveillance and control of invasive mosquito was established [15]. For an efficient vector control at immature stages, accurate identification of mosquito species at the larval stage is compulsory. The molecular biology approach for mosquito identification remains laborious, time-consuming and costly [18]
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