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

Pathogens ◽  
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.

scholarly journals An integrative approach to identify sand fly vectors of leishmaniasis in Ethiopia by morphological and molecular techniques

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Myrthe Pareyn ◽  
Vit Dvorak ◽  
Petr Halada ◽  
Natalie Van Houtte ◽  
Nigatu Girma ◽  
...  
Keyword(s):  
Species Identification ◽  
Molecular Techniques ◽  
Protein Profiling ◽  
Sand Fly ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Human Leishmaniasis ◽  
Sand Fly Vectors ◽  
Morphological Assessment ◽  
Tof Ms

Abstract Background Ethiopia is affected by human leishmaniasis caused by several Leishmania species and transmitted by a variety of sand fly vectors of the genus Phlebotomus. The sand fly fauna in Ethiopia is highly diverse and some species are closely related and similar in morphology, resulting in difficulties with species identification that requires deployment of molecular techniques. DNA barcoding entails high costs, requires time and lacks reference sequences for many Ethiopian species. Yet, proper species identification is pivotal for epidemiological surveillance as species differ in their actual involvement in transmission cycles. Recently, protein profiling using MALDI-TOF mass spectrometry has been introduced as a promising technique for sand fly identification. Methods In our study, we used an integrative taxonomic approach to identify most of the important sand fly vectors of leishmaniasis in Ethiopia, applying three complementary methods: morphological assessment, sequencing analysis of two genetic markers, and MALDI-TOF MS protein profiling. Results Although morphological assessment resulted in some inconclusive identifications, both DNA- and protein-based techniques performed well, providing a similar hierarchical clustering pattern for the analyzed species. Both methods generated species-specific sequences or protein patterns for all species except for Phlebotomus pedifer and P. longipes, the two presumed vectors of Leishmania aethiopica, suggesting that they may represent a single species, P. longipes Parrot & Martin. All three approaches also revealed that the collected specimens of Adlerius sp. differ from P. (Adlerius) arabicus, the only species of Adlerius currently reported in Ethiopia, and molecular comparisons indicate that it may represent a yet undescribed new species. Conclusions Our study uses three complementary taxonomical methods for species identification of taxonomically challenging and yet medically import Ethiopian sand flies. The generated MALDI-TOF MS protein profiles resulted in unambiguous identifications, hence showing suitability of this technique for sand fly species identification. Furthermore, our results contribute to the still inadequate knowledge of the sand fly fauna of Ethiopia, a country severely burdened with human leishmaniasis.

scholarly journals The use of Principle Component Analysis and MALDI-TOF MS for the differentiation of mineral forming Virgibacillus and Bacillus species isolated from Sabkhas

2020 ◽  
Author(s):  
Rim Abdel Samad ◽  
Zulfa Al Disi ◽  
Mohammad Ashfaq ◽  
Nabil Zouari
Keyword(s):  
Clustering Analysis ◽  
Potential Role ◽  
Protein Profiling ◽  
Bacillus Species ◽  
Pca Analysis ◽  
Bacterial Strains ◽  
Protein Biomarkers ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Tof Ms

Occurrence of mineral forming and other bacteria in mats is well demonstrated. However, their high diversity shown by ribotyping was not explained, although it could explain the diversity of formed minerals. Common biomarkers as well as phylogenic relationships are useful tools to clustering the isolates and predict their potential role in the natural niche. In this study, combination of MALDI-TOF MS with PCA was shown a powerful tool to categorize 35 mineral forming bacterial strains isolated from Dohat Fshaikh sabkha, at northwest of Qatar (23 from decaying mats and 12 from living ones). 23 strains from decaying mats belong to Virgibacillus genus as identified by ribotyping and are shown highly involved in formation of protodolomite and a diversity of minerals. They were used as internal references in categorization of sabkha bacteria. Combination of isolation of bacteria on selective mineral forming media, their MALDI TOF MS protein profiling and PCA analysis established their relationship in a phyloproteomic based on protein biomarkers including m/z 4905, 3265, 5240, 6430, 7765, and 9815. PCA analysis clustered the studied strains into 3 major clusters, showing strong correspondence to the 3 phyloproteiomic groups that were established by the dendrogram. Both clustering analysis means have evidently demonstrated a relationship between known Virgibacillus strains and other related bacteria based on profiling of their synthesized proteins. Thus, larger populations of bacteria in mats can be easily screened for their potential to exhibit certain activities, which is of ecological, environmental and biotechnological significance.

scholarly journals Phlebotomine Sand Fly Survey in the Republic of Moldova: Species Composition, Distribution and Host Preferences

2021 ◽  
Author(s):  
Tatiana Sulesco ◽  
Ozge Erisoz Kasap ◽  
Petr Halada ◽  
Gizem Oguz ◽  
Dimian Rusnac ◽  
...  
Keyword(s):  
Species Composition ◽  
Pathogen Transmission ◽  
Molecular Techniques ◽  
Protein Profiling ◽  
Sand Fly ◽  
Sand Flies ◽  
Republic Of Moldova ◽  
Maldi Tof ◽  
Peptide Mass ◽  
The Republic

Abstract Background: Phlebotomine sand flies (Diptera: Psychodiae) in the Republic of Moldova have been understudied for decades. Our study provides a first update on their occurrence, species composition and bloodmeal sources after fifty years.Methods: During 5 seasons (2013-2017), 58 localities from 20 regions were surveyed for presence of sand flies using CDC light traps and manual aspirators. Species identification was done by a combination of morphological and molecular approaches (DNA barcoding, MALDI-TOF MS protein profiling). In engorged females, host blood was identified by three molecular techniques (RFLP, cytb sequencing and MALDI-TOF peptide mass mapping). Population structure of most abundant species was studied by cox1 haplotyping, phylogenetic analyses of ITS2 and cox1 genetic markers were used to resolve relationships of other detected species.Results: In total, 780 sand flies were collected at 30 (51.7%) localities from 12 regions of Moldova. Three species were identified by an integrative morphological and molecular approach: Phlebotomus papatasi, P. perfiliewi and Adlerius sp., the first being the most abundant and widespread, markedly anthropophilic based of bloodmeal analyses and occurring also indoor, showing low structuring of population with only 5 haplotypes of cox1 detected. Distinct morphological and molecular characters of Adlerius sp. specimens suggest presence of yet undescribed species.Conclusions: Our study revealed a presence of stable sand fly population of three species in Moldova that pose a nuisance by biting as well as potential thread of pathogen transmission and shall be further studied.

scholarly journals Effect of trapping method on species identification of phlebotomine sandflies by MALDI-TOF MS protein profiling

2018 ◽  
Vol 32 (3) ◽  
pp. 388-392 ◽  
Author(s):  
P. Halada ◽  
K. Hlavackova ◽  
J. Risueño ◽  
E. Berriatua ◽  
P. Volf ◽  
...  
Keyword(s):  
Species Identification ◽  
Protein Profiling ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Phlebotomine Sandflies ◽  
Trapping Method ◽  
Tof Ms

scholarly journals MALDI-TOF MS protein profiling for the rapid identification of Chagas disease triatomine vectors and application to the triatomine fauna of French Guiana – CORRIGENDUM

Parasitology ◽  
2017 ◽  
Vol 145 (5) ◽  
pp. 676-676 ◽  
Author(s):  
MAUREEN LAROCHE ◽  
JEAN-MICHEL BÉRENGER ◽  
GLADYS GAZELLE ◽  
DENIS BLANCHET ◽  
DIDIER RAOULT ◽  
...  
Keyword(s):  
Chagas Disease ◽  
French Guiana ◽  
Rapid Identification ◽  
Protein Profiling ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Tof Ms

Thymosin β4 is differentially expressed in the cerebrospinal fluid of Creutzfeldt-Jakob disease patients: a MALDI-TOF MS protein profiling study

2009 ◽  
Vol 3 (5) ◽  
pp. 574-583 ◽  
Author(s):  
Antonio Qualtieri ◽  
Elena Urso ◽  
Maria Le Pera ◽  
Sabrina Bossio ◽  
Francesca Bernaudo ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Protein Profiling ◽  
Differentially Expressed ◽  
Thymosin Β4 ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Jakob Disease ◽  
Tof Ms

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

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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