Longitudinal monitoring of environmental factors at Culicidae larval habitats in urban areas and their association with various mosquito species using an innovative strategy

Nebbak, Amira; Willcox, Alexandra C; Koumaré, S; Bérenger, Jean-Michel; Raoult, Didier; Parola, Philippe; Fontaine, Albin; Briolant, Sébastien; Almeras, Lionel

doi:10.1002/ps.5196

Cited by 14 publications

(10 citation statements)

References 40 publications

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“…The phenomena of protein degradation/digestion generating unstable MS profiles, was previously reported when blood meals from engorged mosquitoes were analyzed by MALDI-TOF MS approach [34]. Effectively, the lower protein abundance, for instance, in early larval stages [30] or eggs [35] from mosquitoes conducted to lower MS spectra, thus affecting the rate of reliable identification.…”

Section: Discussionmentioning

confidence: 95%

“…This MS biotyping tool was successfully applied for the identification of mosquitoes at the adult [20,29] but also at the immature stages [21,22]. Nevertheless, the rates of correct identification at the pupal stages were lower than at late-instar stages using MALDI-TOF MS biotyping [22,30]. At the beginning of the pupal cycle, MS profiles are shared with counter species MS spectra from late larval instar stages explaining specimen identification success [22].…”

Section: Discussionmentioning

confidence: 99%

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Identification of Aedes mosquitoes by MALDI-TOF MS biotyping using protein signatures from larval and pupal exuviae

Nebbak

Alméras

2020

Parasites Vectors

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Background: 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 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. Methods:To 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.Results: Reproducible 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. Conclusions:The 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.

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Section: Discussionmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 99%

Identification of Aedes mosquitoes by MALDI-TOF MS biotyping using protein signatures from larval and pupal exuviae

Nebbak

Alméras

2020

Parasites Vectors

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“…As MALDI-TOF MS is well-adapted for high throughput analyses, applying an automatic homogenization procedure limiting handling variations and allowing for testing a large number of samples appears more appropriate. The establishment of an automatized procedure for mosquito larvae homogenisation [29,37] allowed the application of MALDI-TOF MS to monitor mosquito fauna at immature stages for several months [52,53]. The homogenization step was automatized to reduce improper sample grinding and improve the acquisition of high-quality MS spectra.…”

Section: Discussionmentioning

confidence: 99%

Enhanced procedures for mosquito identification by MALDI-TOF MS

Bamou

Costa

Diarra

et al. 2022

Preprint

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Background. The increasing interest in arthropod identification by MALDI-TOF MS conducted to the emergence of heterogenic procedures, including sample preparation and selection of body parts to use. The absence of a consensual strategy hampers direct inter-studies comparisons and brings confusion to new users. Establishing optimized procedures and standardized protocols for mosquito identification by MALDI-TOF MS becomes compulsory, allowing notably the sharing of reference MS databases. Objective. Then, here, we propose to assess the optimal conditions for mosquito identification using MALDI-TOF MS profiling. Methods. Three homogenization methods, two manuals and one automatic, applied on three distinct body parts, legs, thorax and head, were evaluated on two mosquito laboratory strains, Anopheles coluzzii and Aedes aegypti. MS profiles reproducibility, identification rate with relevant scores and the suitable procedures for high-throughput analyses were the main criteria for establishing optimized guidelines. Additionally, the consequences of blood feeding and geographical origin were evaluated using both laboratory strain and field collected mosquitoes. Results. The three body parts exhibited relevant score values for mosquito identification using MALDI-TOF MS profiling; however, thorax and legs appeared the most suitable, independently of homogenization methods or species. Although manual grindings displayed high identification performance for the three body parts, this homogenization mode is not adapted to process a large number of samples. Then, the automatic procedure was selected as the reference homogenization method. Blood feeding status did not hamper the identification of mosquito species, despite the presence of MS peaks from blood origin in MS profiles from three body parts tested on both species. Finally, a significant improvement in identification scores was obtained for specimens from the field when MS spectra of species from the same geographical area were added to the database. Conclusion. The present work established guidelines concerning the selection of mosquito anatomic part and modality of sample preparation (e.g., homogenization) for future specimen identification by MALDI-TOF MS profiling. These standardized operational protocols could be used as references for creating an international MS database.

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“…Recently, MALDI-TOF MS was relevantly applied for arthropods identification, including mosquitoes [ 28 ]. This innovative proteomic tool is now recognized to be compatible with field investigation [ 29 , 30 ].…”

Section: Introductionmentioning

confidence: 99%

Virome Diversity among Mosquito Populations in a Sub-Urban Region of Marseille, France

Nebbak

Monteil-Bouchard

Bérenger

et al. 2021

Viruses

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Some mosquito species have significant public health importance given their ability to transmit major diseases to humans and animals, making them the deadliest animals in the world. Among these, the Aedes (Ae.) genus is a vector of several viruses such as Dengue, Chikungunya, and Zika viruses that can cause serious pathologies in humans. Since 2004, Ae. albopictus has been encountered in the South of France, and autochthonous cases of Dengue, Chikungunya, and Zika diseases have recently been reported, further highlighting the need for a comprehensive survey of the mosquitoes and their associated viruses in this area. Using high throughput sequencing (HTS) techniques, we report an analysis of the DNA and RNA viral communities of three mosquito species Ae. albopictus, Culex (Cx.) pipiens, and Culiseta (Cs.) longiareolata vectors of human infectious diseases in a small sub-urban city in the South of France. Results revealed the presence of a significant diversity of viruses known to infect bacteria, plants, insects, and mammals. Several novel viruses were detected, including novel members of the Rhabdoviridae, Totiviridae, Iflaviviridae, Circoviridae, and Sobemoviridae families. No sequence related to major zoonotic viruses transmitted by mosquitoes was detected. The use of HTS on arthropod vector populations is a promising strategy for monitoring the emergence and circulation of zoonoses and epizooties. This study is a contribution to the knowledge of the mosquito microbiome.

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Longitudinal monitoring of environmental factors at Culicidae larval habitats in urban areas and their association with various mosquito species using an innovative strategy

Cited by 14 publications

References 40 publications

Identification of Aedes mosquitoes by MALDI-TOF MS biotyping using protein signatures from larval and pupal exuviae

Identification of Aedes mosquitoes by MALDI-TOF MS biotyping using protein signatures from larval and pupal exuviae

Enhanced procedures for mosquito identification by MALDI-TOF MS

Virome Diversity among Mosquito Populations in a Sub-Urban Region of Marseille, France

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