scholarly journals Ecology, seasonality and host preferences of Austrian Phlebotomus (Transphlebotomus) mascittii Grassi, 1908, populations

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Edwin Kniha ◽  
Markus Milchram ◽  
Vít Dvořák ◽  
Petr Halada ◽  
Adelheid G. Obwaller ◽  
...  
Keyword(s):  
Barometric Pressure ◽  
Protein Profiling ◽  
Sand Fly ◽  
Sand Flies ◽  
Protozoan Parasites ◽  
Host Preferences ◽  
Maldi Tof ◽  
Increased Activity ◽  
Blood Meals ◽  
Pressure Analysis

Abstract Background Sand flies are principal vectors of the protozoan parasites Leishmania spp. and are widely distributed in all warmer regions of the world, including the Mediterranean parts of Europe. In Central European countries, the sand fly fauna is still under investigation. Phlebotomus mascittii, a suspected but unproven vector of Leishmania infantum, is regarded as the most widely distributed species in Europe. However, many aspects of its biology and ecology remain poorly known. The aim of this study was to provide new data on the biology and ecology of Ph. mascittii in Austria to better understand its current distribution and potential dispersal. Methods Sand flies were collected by CDC light traps at four localities in Austria for 11 (2018) and 15 weeks (2019) during the active sand fly season. Climatic parameters (temperature, relative humidity, barometric pressure and wind speed) were retrospectively obtained for the trapping periods. Sand flies were identified by a combined approach (morphology, DNA barcoding, MALDI-TOF protein profiling), and blood meals of engorged females were analysed by DNA sequencing and MALDI-TOF mass spectrometry. Results In total, 450 individuals of Ph. mascittii were caught. Activity was observed to start at the beginning of June and end at the end of August with peaks in mid-July at three locations and early August at one location. Increased activity was associated with relatively high temperatures and humidity. Also, more individuals were caught on nights with low barometric pressure. Analysis of five identified blood meals revealed chicken (Gallus gallus) and equine (Equus spp.) hosts. Sand fly abundance was generally associated with availability of hosts. Conclusion This study reports unexpectedly high numbers of Ph. mascittii at selected Austrian localities and provides the first detailed analysis of its ecology to date. Temperature and humidity were shown to be good predictors for sand fly activity. Blood meal analyses support the assumption that Ph. mascittii feeds on mammals as well as birds. The study significantly contributes to understanding the ecology of this sand fly species in Central Europe and facilitates prospective entomological surveys.

scholarly journals Ecology, Seasonality and Host Preferences of Austrian Phlebotomus (Transphlebotomus) Mascittii Grassi, 1908, Populations

2021 ◽  
Author(s):  
Edwin Kniha ◽  
Markus Milchram ◽  
Vít Dvořák ◽  
Petr Halada ◽  
Adelheid G. Obwaller ◽  
...  
Keyword(s):  
Barometric Pressure ◽  
Sand Fly ◽  
Sand Flies ◽  
Active Period ◽  
Climatic Parameters ◽  
Protozoan Parasites ◽  
Host Preferences ◽  
Central European ◽  
The World ◽  
Blood Meals

Abstract Background: Sand flies are principal vectors of the protozoan parasites Leishmania spp., widely distributed in all warmer regions of the world, including the Mediterranean parts of Europe. In Central European countries, the sand fly fauna is still understudied. Phlebotomus mascittii, a suspected but unproven vector of Leishmania infantum, is regarded as the most widely distributed species in Europe. However, many aspects of its biology and ecology remain poorly known. The aim of this study was to provide new data on Ph. mascittii biology and ecology in Austria to better understand its current distribution and potential dispersal.Methods: Sand fly trappings were conducted at four different locations in Austria for eleven and fifteen weeks during the active period in 2018 and 2019, respectively. Climatic parameters including temperature, relative humidity, barometric pressure and wind speed were retrospectively obtained for the trapping periods and blood meals of engorged females were analysed by DNA sequencing and MALDI-TOF mass spectrometry.Results: In total, 450 individuals of Ph. mascittii were caught. Activity was observed to start at the beginning of June and end at the end of August with peaks in mid-July at three locations and early August at one location. Increasing activity was associated with relatively high temperatures and humidity. Also, more individuals were caught in nights with low barometric pressure. Five identified blood meals revealed three times chicken (Gallus gallus) and twice equines (Equus spp.) as hosts. Sand fly abundance was generally associated with availability of hosts.Conclusion: This study reports an unexpectedly high abundance of Ph. mascittii at studied Austrian localities and provides the first detailed analysis of its ecology to date. Temperature and humidity were shown to be good predictors for sand fly activity. Blood meal analyses support the assumption that Ph. mascittii feeds on mammals as well as birds. The study significantly contributes to understanding the ecology of this sand fly species in Central Europe and facilitates prospective entomological surveys.

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 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 Phlebotomine sand fly survey in the Republic of Moldova: species composition, distribution and host preferences

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Tatiana Șuleșco ◽  
Ozge Erisoz Kasap ◽  
Petr Halada ◽  
Gizem Oğuz ◽  
Dimian Rusnac ◽  
...  
Keyword(s):  
Population Structure ◽  
Species Composition ◽  
Pathogen Transmission ◽  
Molecular Techniques ◽  
Protein Profiling ◽  
Sand Fly ◽  
Sand Flies ◽  
Republic Of Moldova ◽  
Maldi Tof ◽  
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 50 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, 793 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 Phlebotomus sp. (Adlerius), the first being the most abundant and widespread, markedly anthropophilic based on bloodmeal analyses, occurring also indoors and showing low population structure with only five haplotypes of cox1 detected. Distinct morphological and molecular characters of Phlebotomus sp. (Adlerius) specimens suggest the presence of a yet undescribed species. Conclusions Our study revealed the presence of stable sand fly populations of three species in Moldova that represent a biting nuisance as well as a potential threat of pathogen transmission and shall be further studied. Graphical abstract

scholarly journals Sand fly fauna of Crete and the description of Phlebotomus (Adlerius) creticus n. sp. (Diptera: Psychodidae)

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Vít Dvořák ◽  
Nikolaos Tsirigotakis ◽  
Christoforos Pavlou ◽  
Emmanouil Dokianakis ◽  
Mohammad Akhoundi ◽  
...  
Keyword(s):  
Field Work ◽  
Abundant Species ◽  
Protein Profiling ◽  
Sand Fly ◽  
Published Data ◽  
Specific Sequence ◽  
Maldi Tof ◽  
Morphological Criteria ◽  
Species Specific ◽  
Protein Spectra

Abstract Background The Greek island of Crete is endemic for both visceral leishmaniasis (VL) and recently increasing cutaneous leishmaniasis (CL). This study summarizes published data on the sand fly fauna of Crete, the results of new sand fly samplings and the description of a new sand fly species. Methods All published and recent samplings were carried out using CDC light traps, sticky traps or mouth aspirators. The specific status of Phlebotomus (Adlerius) creticus n. sp., was assessed by morphological analysis, cytochrome b (cytb) sequencing and MALDI-TOF protein profiling. Results Published data revealed the presence of 10 Phlebotomus spp. and 2 Sergentomyia spp. During presented field work, 608 specimens of 8 species of Phlebotomus and one species of Sergentomyia were collected. Both published data and present samplings revealed that the two most common and abundant species were Phlebotomus neglectus, a proven vector of Leishmania infantum causing VL, and Ph. similis, a suspected vector of L. tropica causing CL. In addition, the field surveys revealed the presence of a new species, Ph. (Adlerius) creticus n. sp. Conclusions The identification of the newly described species is based on both molecular and morphological criteria, showing distinct characters of the male genitalia that differentiate it from related species of the subgenus Adlerius as well as species-specific sequence of cytb and protein spectra generated by MALDI-TOF mass spectrometry.

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.

Identification of mixed and successive blood meals of mosquitoes using MALDI-TOF MS protein profiling

Parasitology ◽  
2020 ◽  
Vol 147 (3) ◽  
pp. 329-339 ◽  
Author(s):  
Fatalmoudou Tandina ◽  
Sirama Niare ◽  
Lionel Almeras ◽  
Bernard Davoust ◽  
Ogobara K Doumbo ◽  
...  
Keyword(s):  
Protein Profiling ◽  
Correct Identification ◽  
Anopheles Coluzzii ◽  
Blind Test ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Mixed Blood ◽  
Vertebrate Hosts ◽  
Blood Meals ◽  
Tof Ms

AbstractBackgroundThe accurate and rapid identification of mosquito blood meals is critical to study the interactions between vectors and vertebrate hosts and, subsequently, to develop vector control strategies. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) profiling has been shown to be a reliable and effective tool for identifying single blood meals from mosquitoes.MethodsIn this study, we developed MALDI-TOF MS profiling protocols to identify Anopheles gambiae Giles, Anopheles coluzzii and Aedes albopictus mosquitoes’ mixed blood meals and the last of successive blood meals. The mosquitoes were either successively artificially fed with distinct host bloods or engorged with mixed bloods from distinct vertebrate hosts, such as humans, sheep and dogs.ResultsBlind test analyses revealed a correct identification of mixed blood meals from mosquitoes using MALDI-TOF MS profiling. The 353 MS spectra from mixed blood meals were identified using log score values >1.8. All MS spectra (n = 244) obtained from mosquitoes' successive blood meals were reproducible and specific to the last blood meal, suggesting that the previous blood meals do not have an impact on the identification of the last one.ConclusionMALDI-TOF MS profiling approach appears to be an effective and robust technique to identify the last and mixed blood meals during medical entomological surveys.

scholarly journals Second Blood Meal by Female Lutzomyia longipalpis: Enhancement by Oviposition and Its Effects on Digestion, Longevity, and Leishmania Infection

2018 ◽  
Vol 2018 ◽  
pp. 1-10 ◽  
Author(s):  
C. S. Moraes ◽  
K. Aguiar-Martins ◽  
S. G. Costa ◽  
P. A. Bates ◽  
R. J. Dillon ◽  
...  
Keyword(s):  
Blood Meal ◽  
Molecular Mechanisms ◽  
Sand Fly ◽  
Leishmania Infection ◽  
Lutzomyia Longipalpis ◽  
Sand Flies ◽  
Gonotrophic Cycle ◽  
Trypsin Activity ◽  
The Impact ◽  
Blood Meals

Lutzomyia longipalpis is the main vector of visceral leishmaniasis (VL) in America. Physiological and molecular mechanisms of Leishmania infection in sand flies have been studied during the first gonotrophic cycle. There are few studies about these interactions during the second gonotrophic cycle mainly because of the difficulties maintaining sand flies through sequential feeds. Here we standardized conditions to perform the second blood feed efficiently, and our results show that oviposition is an essential factor for the success of multiple feeds. We evaluated the impact of the second blood meal on longevity, protein digestion, trypsin activity, and Leishmania mexicana development within L. longipalpis gut. Mortality of blood-fed females increases after second blood meal as compared to sugar-fed females. Trypsin activity was lower during the second gonotrophic cycle. However, no difference in protein intake was observed between blood meals. There was no difference in the population size of Leishmania in the gut after both blood meals. In this work, we presented an optimized protocol for obtaining sufficient numbers of sand fly females fed on a second blood meal, and we described some physiological and parasitological aspects of the second gonotrophic cycle which might influence the vectorial competence of sand flies.

scholarly journals Molecular detection of Leishmania donovani, Leishmania major, and Trypanosoma species in Sergentomyia squamipleuris sand flies from a visceral leishmaniasis focus in Merti sub-County, eastern Kenya

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Barrack O. Owino ◽  
Jackline Milkah Mwangi ◽  
Steve Kiplagat ◽  
Hannah Njiriku Mwangi ◽  
Johnstone M. Ingonga ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Mus Musculus ◽  
Blood Meal ◽  
Leishmania Donovani ◽  
Leishmania Major ◽  
Sand Fly ◽  
Health Concern ◽  
Sand Flies ◽  
Reservoir Hosts ◽  
Blood Meals

Abstract Background Visceral leishmaniasis (VL) and zoonotic cutaneous leishmaniasis (ZCL) are of public health concern in Merti sub-County, Kenya, but epidemiological data on transmission, vector abundance, distribution, and reservoir hosts remain limited. To better understand the disease and inform control measures to reduce transmission, we investigated the abundance and distribution of sand fly species responsible for Leishmania transmission in the sub-County and their blood-meal hosts. Methods We conducted an entomological survey in five villages with reported cases of VL in Merti sub-County, Kenya, using CDC miniature light traps and castor oil sticky papers. Sand flies were dissected and identified to the species level using standard taxonomic keys and PCR analysis of the cytochrome c oxidase subunit 1 (cox1) gene. Leishmania parasites were detected and identified by PCR and sequencing of internal transcribed spacer 1 (ITS1) genes. Blood-meal sources of engorged females were identified by high-resolution melting analysis of vertebrate cytochrome b (cyt-b) gene PCR products. Results We sampled 526 sand flies consisting of 8 species, Phlebotomus orientalis (1.52%; n = 8), and 7 Sergentomyia spp. Sergentomyia squamipleuris was the most abundant sand fly species (78.71%; n = 414) followed by Sergentomyia clydei (10.46%; n = 55). Leishmania major, Leishmania donovani, and Trypanosoma DNA were detected in S. squamipleuris specimens. Humans were the main sources of sand fly blood meals. However, we also detected mixed blood meals; one S. squamipleuris specimen had fed on both human and mouse (Mus musculus) blood, while two Ph. orientalis specimens fed on human, hyrax (Procavia capensis), and mouse (Mus musculus) blood. Conclusions Our findings implicate the potential involvement of S. squamipleuris in the transmission of Leishmania and question the dogma that human leishmaniases in the Old World are exclusively transmitted by sand flies of the Phlebotomus genus. The presence of Trypanosoma spp. may indicate mechanical transmission, whose efficiency should be investigated. Host preference analysis revealed the possibility of zoonotic transmission of leishmaniasis and other pathogens in the sub-County. Leishmania major and L. donovani are known to cause ZCL and VL, respectively. However, the reservoir status of the parasites is not uniform. Further studies are needed to determine the reservoir hosts of Leishmania spp. in the area.

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