Hymenopteran Parasitoids of Hard Ticks in Western Africa and the Russian Far East

Some parasitoids of the genus Ixodiphagus (Hymenoptera, Chalcidoidea: Encyrtidae) are well-known natural enemies of ticks. In this study, we investigate the occurrence of parasitoid wasps in adult hard ticks from Western Africa (Côte d’Ivoire and Senegal) and Far Eastern Europe (Russia) using molecular methods. The morphological identification allowed the classification of 785 collected specimens of six species of ticks: Rhipicephalus (Boophilus) microplus (41%), Ixodes persulcatus (33%), Dermacentor silvarum (11%), Haemaphysalis concinna (7%), Amblyomma variegatum (5%), and Haemaphysalis japonica (3%). The newly developed MALDI-TOF MS protocol identified tick species in spite of their different storage (dried or in 70% ethanol) conditions for a long period. Molecular screening of ticks by a new standard PCR system developed in this study revealed the presence of parasitoid wasp DNA in 3% (28/785) of analyzed ticks. Ixodiphagus hookeri was detected in 86% (24/28) of infested ticks, including 13 I. persulcatus, 9 R (B) microplus, and one H. concinna and D. silvarum. While an unidentified parasitoid wasp species from the subfamily Aphidiinae and Braconidae family was detected in the remaining 14% (4/28) infested ticks. These infested ticks were identified as I. persulcatus. Our findings highlight the need for further studies to clarify the species diversity of parasitoid infesting ticks by combining molecular and morphological features. The novel molecular and MALDI-TOF MS protocols could be effective tools for the surveillance and characterization of these potential bio-control agents of ticks.

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Discrimination between Asian populations of the parasitoid wasp Ganaspis cf. brasiliensis using a simple MALDI-TOF MS-based method for use with insects

Biology Methods and Protocols ◽

10.1093/biomethods/bpz002 ◽

2019 ◽

Vol 4 (1) ◽

Cited By ~ 3

Author(s):

Michael A Reeve ◽

M Lukas Seehausen

Keyword(s):

Reference Sample ◽

Principal Component ◽

Parasitoid Wasp ◽

Test Sample ◽

Invasive Pest ◽

Drosophila Suzukii ◽

Maldi Tof Ms ◽

Chinese Populations ◽

Maldi Tof ◽

Tof Ms

Abstract The fruit fly Drosophila suzukii has recently become an invasive pest insect of significant economic impact in Europe and the USA. In contrast to other Drosophila species, D. suzukii is able to infest intact fruit by means of a saw-like ovipositor, which allows females to deposit eggs beneath the skin of the fruit. Classical biological control using the parasitoid wasp Ganaspis cf. brasiliensis is currently being researched as an environmentally sustainable option for the control of D. suzukii. In particular, the host specificity of this parasitoid has been assessed for populations from different regions in China and Japan. In order to study the relationship between the differences in specificity and molecular variations, we have adapted a matrix-assisted laser-desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS)-based method, originally developed for use with plant material, to discriminate between example populations of G. cf. brasiliensis. We have employed a combination of principal component analysis and blind-tested comparison between reference sample MALDI-TOF MS spectra and test sample spectra to discriminate, on the basis of the acid-soluble insect protein spectra generated, between four populations of G. cf. brasiliensis (originally collected from Tokyo and Hasuike in Japan and Dali and Ximing in China). MALDI-TOF MS analysis is able to discriminate with 100% accuracy between populations G. cf. brasiliensis. The Chinese populations were observed to be similar, but the Tokyo population is slightly different and the Hasuike population is significantly different from the other populations. The Tokyo population appears more closely related to the Chinese populations than the Hasuike population, even though both originate from Japan.

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Using MALDI-TOF mass spectrometry to identify ticks collected on domestic and wild animals from the Democratic Republic of the Congo

Experimental and Applied Acarology ◽

10.1007/s10493-021-00629-z ◽

2021 ◽

Author(s):

Steve Ngoy ◽

Adama Zan Diarra ◽

Anne Laudisoit ◽

Guy-Crispin Gembu ◽

Erik Verheyen ◽

...

Keyword(s):

Mass Spectrometry ◽

Democratic Republic ◽

Morphological Identification ◽

Ionization Mass ◽

Wild Animals ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Republic Of The Congo ◽

Reference Specimens ◽

Tof Ms

AbstractMatrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS) has recently emerged as an alternative to morphological and molecular tools to identify tick species. In this study, we set out to evaluate and confirm the ability of MALDI-TOF MS to identify different species of ticks collected in the Democratic Republic of the Congo and preserved in 70% ethanol. A total of 575 ticks, of which 530 were collected from domestic pigs and 45 from wild animals, were subjected to MALDI-TOF MS analysis to evaluate the intraspecies reproducibility and interspecies specificity of MS profiles obtained from the different species. Morphologically, the ticks belonged to seven different species, namely Rhipicephalus complanatus, Rhipicephalus congolensis, Haemaphysalis muhsamae, Ixodes cumulatimpunctatus, Amblyomma exornatum, Amblyomma compressum and an unidentified Rhipicephalus sp. A total of 535/575 (93%) of the spectra obtained were of good enough quality to be used for our analyses. Our home-made MALDI-TOF MS arthropod database was upgraded with spectra obtained from between one and five randomly selected specimens per species. For these reference specimens, molecular identification of the ticks was also made using 16S, 12S rDNA genes and the Cox1 mtDNA gene sequencing. The remaining good quality spectra were then queried against the upgraded MALDI-TOF MS database, showing that 100% were in agreement with the morphological identification, with logarithmic score values (LSVs) between 1.813 and 2.51. The consistency between our morphological, molecular and MALDI-TOF MS identification confirms the capability and precision of MALDI-TOF MS for tick identification.

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Molecular and MALDI-TOF MS identification of swallow bugs Cimex hirundinis (Heteroptera: Cimicidae) and endosymbionts in France

Parasites & Vectors ◽

10.1186/s13071-021-05073-x ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Fatima Zohra Hamlili ◽

Jean-Michel Bérenger ◽

Adama Zan Diarra ◽

Philippe Parola

Keyword(s):

Developmental Stages ◽

Blood Feeding ◽

Rapid Identification ◽

Morphological Identification ◽

Blind Test ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Minimal Sample ◽

House Martin ◽

Tof Ms

Abstract Background The Cimicidae are obligatory blood-feeding ectoparasites of medical and veterinary importance. We aim in the current study to assess the ability of MALDI-TOF MS to identify Cimex hirundinis swallow bugs collected in house martin nests. Methods Swallow bugs were picked out from abandoned nests of house martin swallows and identified morphologically to the species level. The bugs were randomly selected, dissected and then subjected to MALDI-TOF MS and molecular analyses. Results A total of 65 adults and 50 nymphs were used in the attempt to determine whether this tool could identify the bug species and discriminate their developmental stages. Five adults and four nymphs of C. hirundinis specimens were molecularly identified to update our MS homemade arthropod database. BLAST analysis of COI gene sequences from these C. hirundinis revealed 98.66–99.12% identity with the corresponding sequences of C. hirundinis of the GenBank. The blind test against the database supplemented with MS reference spectra showed 100% (57/57) C. hirundinis adults and 100% (46/46) C. hirundinis nymphs were reliably identified and in agreement with morphological identification with logarithmic score values between 1.922 and 2.665. Ninety-nine percent of C. hirundinis specimens tested were positive for Wolbachia spp. The sequencing results revealed that they were identical to Wolbachia massiliensis, belonging to the new T-supergroup strain and previously isolated from C. hemipterus. Conclusions We report for the first time to our knowledge a case of human infestation by swallow bugs (C. hirundinis) in France. We also show the usefulness of MALDI-TOF MS in the rapid identification of C. hirundinis specimens and nymphs with minimal sample requirements. We phylogenetically characterized the novel Wolbachia strain (W. massiliensis) infecting C. hirundinis and compared it to other recognized Wolbachia clades. Graphical Abstract

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Integrative Approach to Phlebotomus mascittii Grassi, 1908: First Record in Vienna with New Morphological and Molecular Insights

Pathogens ◽

10.3390/pathogens9121032 ◽

2020 ◽

Vol 9 (12) ◽

pp. 1032

Author(s):

Edwin Kniha ◽

Vít Dvořák ◽

Petr Halada ◽

Markus Milchram ◽

Adelheid G. Obwaller ◽

...

Keyword(s):

First Record ◽

Protein Profiling ◽

Sand Fly ◽

Integrative Approach ◽

Marker Genes ◽

Morphological Identification ◽

Sand Flies ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Tof Ms

Sand flies (Diptera: Psychodidae: Phlebotominae) are blood-feeding insects that transmit the protozoan parasites Leishmania spp. and various arthropod-borne (arbo) viruses. While in Mediterranean parts of Europe the sand fly fauna is diverse, in Central European countries including Austria mainly Phlebotomus mascittii is found, an assumed but unproven vector of Leishmania infantum. To update the currently understudied sand fly distribution in Austria, a sand fly survey was performed and other entomological catches were screened for sand flies. Seven new trapping locations of Ph. mascittii are reported including the first record in Vienna, representing also one of the first findings of this species in a city. Morphological identification, supported by fluorescence microscopy, was confirmed by two molecular approaches, including sequencing and matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) protein profiling. Sand fly occurrence and activity were evaluated based on surveyed locations, habitat requirements and climatic parameters. Moreover, a first comparison of European Ph. mascittii populations was made by two marker genes, cytochrome c oxidase subunit 1 (COI), and cytochrome b (cytb), as well as MALDI-TOF mass spectra. Our study provides new important records of Ph. mascittii in Austria and valuable data for prospective entomological surveys. MALDI-TOF MS protein profiling was shown to be a reliable tool for differentiation between sand fly species. Rising temperatures and globalization demand for regular entomological surveys to monitor changes in species distribution and composition. This is also important with respect to the possible vector competence of Ph. mascittii.

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Tabanids as possible pathogen vectors in Senegal (West Africa)

Parasites & Vectors ◽

10.1186/s13071-020-04375-w ◽

2020 ◽

Vol 13 (1) ◽

Author(s):

Mohamed Lamine Keita ◽

Hacène Medkour ◽

Masse Sambou ◽

Handi Dahmana ◽

Oleg Mediannikov

Keyword(s):

West Africa ◽

Pathogen Detection ◽

Leishmania Donovani ◽

Pcr Analysis ◽

Morphological Identification ◽

Setaria Digitata ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Human And Animal Diseases ◽

Tof Ms

Abstract Background Species of the Tabanidae are potent vectors of human and animal diseases, but they have not been thoroughly investigated to date. In Senegal (West Africa), little information is available on these dipterans. Our objective in this study was to investigate Senegalese tabanids and their diversity by using molecular and proteomics approaches, as well as their associated pathogens. Methods A total of 171 female tabanids were collected, including 143 from Casamance and 28 from Niokolo-Koba. The samples were identified morphologically by PCR sequencing and by MALDI-TOF MS, and PCR analysis was employed for pathogen detection and blood-meal characterization. Results The morphological identification revealed four species concordantly with the molecular identification: Atylotus fuscipes (79.5%), Tabanus guineensis (16.4%), Chrysops distinctipennis (3.5%) and Tabanus taeniola (0.6%) (not identified by PCR). The molecular investigation of pathogens revealed the presence of Trypanosoma theileri (6.6%), Leishmania donovani (6.6%), Setaria digitata (1.5%), Rickettsia spp. (5.1%) and Anaplasmataceae bacteria (0.7%) in A. fuscipes. Tabanus guineensis was positive for L. donovani (35.7%), S. digitata (3.6%) and Anaplasmataceae (17.8%). Leishmania donovani has been detected in 50% of C. distinctipennis specimens and the only T. taeniola specimen. No Piroplasmida, Mansonella spp. or Coxeilla burnetii DNA was detected. In addition to humans (96.43%), Chlorocebus sabeus, a non-human primate, has been identified as a host of (3.57%) analysed tabanids. MALDI-TOF MS enabled us to correctly identify all tabanid species that had good quality spectra and to create a database for future identification. Conclusions Tabanids in Senegal could be vectors of several pathogens threatening animal and public health. To fully characterize these dipterans, it is therefore necessary that researchers in entomology and infectiology employ molecular characterization and mass spectrometric techniques such as MALDI-TOF MS to analyse these dipterans in Senegal and West Africa.

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ITS rDNA Gene Analysis Versus MALDI-TOF MS For Identification of Neoscytalidium dimidiatum Isolated from Onychomycosis and Dermatomycosis Cases in Medellin (Colombia)

Microorganisms ◽

10.3390/microorganisms7090306 ◽

2019 ◽

Vol 7 (9) ◽

pp. 306 ◽

Cited By ~ 3

Author(s):

Flórez-Muñoz ◽

Gómez-Velásquez ◽

Loaiza-Díaz ◽

Soares ◽

Santos ◽

...

Keyword(s):

Etiologic Agent ◽

Species Level ◽

Morphological Identification ◽

Reference Spectrum ◽

Nuclear Rdna ◽

Maldi Tof Ms ◽

Neoscytalidium Dimidiatum ◽

Maldi Tof ◽

Tof Ms

Within the Neoscytalidium genus, N. dimidiatum, N. oculus, N. orchidacearum, and N. novaehollandiae have been recognized. Although these species are frequently found in soil, N. dimidiatum has been identified as an etiologic agent of onychomycosis or dermatomycosis, and N. oculus has been identified as an etiologic agent of an ocular lesion. All these species can be cultured in vitro, but their morphological identification by macroscopic and microscopic traits is difficult and imprecise due to their similarity. In this study, 34 isolates of Neoscytalidium spp. from 32 onychomycosis and two dermatomycosis cases in Medellin (Colombia) were identified at the species level using sequencing of the ITS1+5.8S+ITS2 nuclear rDNA region and MALDI-TOF mass spectrometry (MS). Neoscytalidium dimidiatum strain MUM 17.21 was used to construct the reference spectrum in the in-house library to identify the clinical isolates by MALDI-TOF MS. Additionally, N. dimidiatum PPC-216 and PLAB-055 strains were used to validate the in-house constructed reference spectra. Although four groups were observed in the dendrogram obtained from the proteins of each isolate profile, MALDI-TOF MS and sequencing results are in accordance, since all isolates were identified as N. dimidiatum.

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Comparison of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry and Molecular Biology Techniques for Identification of Culicoides (Diptera: Ceratopogonidae) Biting Midges in Senegal

Journal of Clinical Microbiology ◽

10.1128/jcm.01855-14 ◽

2014 ◽

Vol 53 (2) ◽

pp. 410-418 ◽

Cited By ~ 31

Author(s):

Masse Sambou ◽

Maxence Aubadie-Ladrix ◽

Florence Fenollar ◽

Becaye Fall ◽

Hubert Bassene ◽

...

Keyword(s):

Mass Spectrometry ◽

Species Level ◽

Morphological Identification ◽

Biting Midges ◽

Ionization Time ◽

Content Type ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Flight Mass Spectrometry ◽

Tof Ms

Biting midges of the genusCulicoidesare implicated as vectors for a wide variety of pathogens. The morphological identification of these arthropods may be difficult because of a lack of detailed investigation of taxonomy for this species in Africa. However, matrix-assisted laser desorption ionization−time of flight mass spectrometry (MALDI-TOF MS) profiling is efficient for arthropod identification at the species level. This study established a spectrum database ofCulicoidesspp. from Senegal using MALDI-TOF. Identification ofCulicoidesinsects to the species level before mass spectrometry was performed on the basis of morphological characters. MALDI-TOF MS reference spectra were determined for 437 field-caughtCulicoidesof 10 species. The protein profiles of all testedCulicoidesrevealed several peaks with mass ranges of 2 to 20 kDa. In a validation study, 72Culicoidesspecimens in the target species were correctly identified at the species level with a similarity of 95 to 99.9%. FourCulicoidesprotein profiles were misidentified. Nevertheless, six SuperSpectra (C. imicola,C. enderleini,C. oxystoma,C. kingi,C. magnus, andC. fulvithorax) were created. Abdomens of midges were used to amplify and sequence a portion of the mitochondrial cytochrome oxidase I gene (COI). The results obtained using the MALDI-TOF MS method were consistent with the morphological identification and similar to the genetic identification. Protein profiling using MALDI-TOF is an efficient approach for the identification ofCulicoidesspp., and it is economically advantageous for approaches that require detailed and quantitative information of vector species that are collected in field. The database of AfricanCulicoidesMS spectra created is the first database in Africa. The COI sequences of fiveCulicoidesspecies that were previously noncharacterized using molecular methods were deposited in GenBank.

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