Sand flies, which transmit diseases like leishmaniases, bartonellosis, and certain viruses, pose a significant public health threat. Our research focuses on the immune responses of Lutzomyia longipalpis, the primary vector for visceral leishmaniasis in the Americas. We use L. longipalpis LL5 cells as a model to study how sand flies respond to pathogens. These cells exhibit robust immune reactions, producing molecules mainly regulated by the Toll, IMD, Jak-STAT, and RNAi pathways. In previous studies, we detected a non-specific antiviral response in LL5 cells following double-stranded RNAs (dsRNAs) transfection. A previous complete secretome of these cells showed molecules resembling an interferon-like antiviral response when transfected with polyinosinic-polycytidylic acid (poly I:C), a synthetic dsRNA analog. In the current study, we analyzed soluble proteins secreted by LL5 cells after poly I:C transfection. Using comparative mass spectrometry, we examined protein composition of conditioned media depleted of exosomes at 24 h and 48 h. Most proteins uniquely expressed in the transfected groups had low abundance compared to the overall expressed proteins. Interactome prediction analysis revealed that at 24 h, the proteins uniquely found in the secretome of the transfected group were involved in RNA degradation and purine metabolism, while at 48 h they were linked to ribosomal proteins and signaling pathways such as Hedgehog, Transforming Growth Factor-beta (TGF-β), and Wingless/integrated (Wnt). We highlight increased abundance of the TGF-β-induced protein ig-h3 (24 h and 48 h), a Toll-like receptor 3 (48 h), and a hemocytin (48 h) in the secretion of transfected groups compared to the controls. We also performed an interaction analysis of proteins more secreted by the treated group at 24 h and 48 h. Unlike the interactome of uniquely identified proteins, few interactions were observed at 24 h, with a predominance of extracellular matrix and cell adhesion proteins. The set of proteins more secreted at 48 h presented more interactions than at 24 h, with emphasis on catabolic processes, including RNA degradation. These findings indicate that poly I:C transfection in LL5 cells induces the secretion of proteins involved in cellular defense and repair, revealing molecules involved in the LL5 non-specific antiviral response.

• Klíčová slova
• RNA degradation, cell repair, non-specific antiviral response, poly I:C, sand fly cell line, secreted proteins,
• MeSH
• buněčné linie MeSH
• dvouvláknová RNA * genetika   MeSH
• hmyzí proteiny * metabolismus   MeSH
• poly I-C MeSH
• proteom * analýza   MeSH
• proteomika MeSH
• Psychodidae * imunologie   metabolismus   genetika   MeSH
• sekretom * MeSH
• signální transdukce MeSH
• transfekce MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• dvouvláknová RNA * MeSH
• hmyzí proteiny * MeSH
• poly I-C MeSH
• proteom * MeSH

In Europe, sand flies (Diptera: Psychodidae: Phlebotominae) are characteristic Mediterranean fauna, though some species expand their range further north. However, the sand fly fauna of Central Europe remains underreported, particularly in Hungary where recent data is lacking due to limited and outdated entomological surveys. To address this gap, a series of sand fly surveys were conducted in Hungary, with significant findings from two trapping efforts in 2017 and 2024. In 2017, only a single female Phlebotomus papatasi was trapped in northern Hungary, which marks one of the northernmost records of the species. In 2024, a more extensive and geographically wider survey recorded 264 sand flies at 34 sites, including three species: Ph. mascittii, Ph. neglectus, and Ph. papatasi. Sand flies were found across diverse environmental settings, including urban, agricultural, and natural habitats. Particularly, the previously rare presence of Ph. mascittii at rural sites (natural rock formations) was reported. Analysis of historical and current data revealed the presence of four sand fly species in Central and South Transdanubia, with evidence suggesting potential range expansion. Blood meal analysis of engorged females identified a variety of domestic and wild host species, but no Leishmania or Phlebovirus infections were detected. Habitat modelling and linear discriminant analysis indicated substantial climate suitability across Southeast Europe, with most positive sand fly observations observed in discontinuous urban fabric CORINE Land Cover classes. This study offers important insights into the ecology, distribution, and climatic preferences of sand flies in Hungary and provides crucial baseline data to monitor potential future spread.

• Klíčová slova
• Barcoding, Blood meal, Central Europe, Ecology, MALDI-MS, Sand flies,
• Publikační typ
• časopisecké články MeSH

BACKGROUND: We evaluated various membranes for blood-feeding in nine sand fly species from different genera and subgenera. Most of these species are vectors of human-pathogenic Leishmania, whereas Sergentomyia minuta is a herpetophilic sand fly species and a proven vector of Leishmania (Sauroleishmania) tarentolae. METHODS: Female sand flies were offered blood through a range of membranes (chicken, reptilian, and frog skin; synthetic collagen; pig intestine; and duck foot webbing). Two feeding systems (glass feeder, Hemotek) and different blood sources (human, ovine, avian, and reptilian) were used. Feeding trials were conducted under varying thermal and light conditions to determine the optimal parameters. RESULTS: Among the 4950 female S. minuta tested, only a negligible fraction took a blood meal: 2% of the females fed on avian blood, and 0.2% of the females fed on human blood. In eight other species, the chicken membrane was generally more effective than synthetic membranes or pig intestines. For example, Phlebotomus duboscqi refused synthetic membranes, while Lutzomyia longipalpis and P. perniciosus avoided both synthetic membranes and pig intestines. The most effective membrane was duck foot webbing, with four species feeding more readily through it than through the chicken membrane. Additionally, applying coagulated blood plasma to the outer surface of chicken or synthetic membranes significantly increased feeding rates. CONCLUSIONS: Female S. minuta did not reliably feed on blood through the tested membranes, preventing laboratory infection experiments from confirming their vector competence for human-pathogenic Leishmania. However, for future experimental infections of other sand fly species, duck foot webbing has emerged as an effective membrane, and the application of blood plasma to the exterior of membranes may increase the feeding rates.

• Klíčová slova
• Leishmania, Lutzomyia, Phlebotomus, Sergentomyia minuta, Artificial feeding, Vector competence,
• MeSH
• krev * MeSH
• lidé MeSH
• membrány MeSH
• prasata MeSH
• Psychodidae * fyziologie   MeSH
• stravovací zvyklosti * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

In Uzbekistan, the number of reported leishmaniasis cases is rising at the alarming rate. In this work, we studied the phlebotomine sand fly (Diptera: Phlebotominae) diversity in the foci of cutaneous leishmaniasis in the Surxondaryo Region of Uzbekistan and compared it with the data obtained for the same area 50 years ago, when infection prevalence was reportedly low. We found that the implicated vector for zoonotic leishmaniasis, P. papatasi, remained eudominant; the proportion of implicated anthroponotic leishmaniasis vector, P. sergenti, rose significantly from averaged 5.4 to 41.4%; Phlebotomus alexandri, a suspected visceral leishmaniasis vector, was eudominant at two sites, and a second suspected vector for this disease, P. longiductus, was newly recorded in the region. We conclude that the increase in the documented cases of cutaneous leishmaniasis in the Surxondaryo Region of Uzbekistan may be connected to the changes in fauna of sand flies vectoring Leishmania spp.

• Klíčová slova
• Diversity, Leishmania, Phlebotomus,
• MeSH
• hmyz - vektory MeSH
• leishmanióza kožní * epidemiologie   MeSH
• leishmanióza viscerální * epidemiologie   MeSH
• Phlebotomus * MeSH
• Psychodidae * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Uzbekistán epidemiologie   MeSH

Phlebotomine sand flies (Diptera: Phlebotominae) are the principal vectors of Leishmania spp. (Kinetoplastida: Trypanosomatidae). In Central Europe, Phlebotomus mascittii is the predominant species, but largely understudied. To better understand factors driving its current distribution, we infer patterns of genetic diversity by testing for signals of population expansion based on two mitochondrial genes and model current and past climate and habitat suitability for seven post-glacial maximum periods, taking 19 climatic variables into account. Consequently, we elucidate their connections by environmental-geographical network analysis. Most analyzed populations share a main haplotype tracing back to a single glacial maximum refuge area on the Mediterranean coasts of South France, which is supported by network analysis. The rapid range expansion of Ph. mascittii likely started in the early mid-Holocene epoch until today and its spread possibly followed two routes. The first one was through northern France to Germany and then Belgium, and the second across the Ligurian coast through present-day Slovenia to Austria, toward the northern Balkans. Here we present a combined approach to reveal glacial refugia and post-glacial spread of Ph. mascittii and observed discrepancies between the modelled and the current known distribution might reveal yet overlooked populations and potential further spread.

• MeSH
• hmyz - vektory genetika   MeSH
• Leishmania * MeSH
• Phlebotomus * genetika   MeSH
• Psychodidae * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Evropa MeSH

Phlebotomine sand flies (Diptera: Psychodidae) are the principal vectors of phleboviruses and Leishmania spp., the causative agents of leishmaniases. The Mediterranean sand fly fauna is diverse, and leishmaniasis, mainly caused by Leishmania infantum, is endemic in the Balkan countries. Despite recent entomological surveys, only some districts of Kosovo have been sampled for sand flies, with no proof/confirmation of L. infantum. This study aimed to gain further insights into the species composition of natural sand fly populations in previously unsampled districts and areas in Kosovo without reports of leishmaniasis and to detect Leishmania DNA in sand flies. A sand fly survey was conducted in 2022 in all seven districts of Kosovo. Collected females were screened for Leishmania DNA by PCR. Positive samples were sequenced and subjected to maximum likelihood analysis with reference sequences for further molecular characterization. The trapping activities at 114 different localities resulted in 3272 caught specimens, comprising seven sand fly species of two genera, namely Phlebotomus neglectus, Ph. perfiliewi, Ph. tobbi, Ph. papatasi, Ph. simici, Ph. balcanicus and Sergentomyia minuta. Leishmania infantum DNA was detected in three individual sand flies of Ph. neglectus and Ph. perfiliewi. This study provides the most extensive sand fly survey in Kosovo and reports the first record of L. infantum DNA in sand flies, indicating autochthonous circulation of L. infantum.

• Klíčová slova
• Balkan, Kosovo, Larroussius, PCR, Phlebotominae, leishmaniasis,
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Phlebotomine sand flies (Diptera: Psychodidae) are important vectors of various human and animal pathogens such as Bartonella bacilliformis, Phlebovirus, and parasitic protozoa of the genus Leishmania, causative agent of leishmaniases that account among most significant vector-borne diseases. The Maghreb countries Mauritania, Morocco, Algeria, Tunisia, and Libya occupy a vast area of North Africa and belong to most affected regions by these diseases. Locally varying climatic and ecological conditions support diverse sand fly fauna that includes many proven or suspected vectors. The aim of this review is to summarize often fragmented information and to provide an updated list of sand fly species of the Maghreb region with illustration of species-specific morphological features and maps of their reported distribution. MATERIALS AND METHODS: The literature search focused on scholar databases to review information on the sand fly species distribution and their role in the disease transmissions in Mauritania, Morocco, Algeria, Tunisia, and Libya, surveying sources from the period between 1900 and 2020. Reported distribution of each species was collated using Google Earth, and distribution maps were drawn using ArcGIS software. Morphological illustrations were compiled from various published sources. RESULTS AND CONCLUSIONS: In total, 32 species of the genera Phlebotomus (Ph.) and Sergentomyia (Se.) were reported in the Maghreb region (15 from Libya, 18 from Tunisia, 23 from Morocco, 24 from Algeria, and 9 from Mauritania). Phlebotomus mariae and Se. africana subsp. asiatica were recorded only in Morocco, Ph. mascitti, Se. hirtus, and Se. tiberiadis only in Algeria, whereas Ph. duboscqi, Se. dubia, Se. africana africana, Se. lesleyae, Se. magna, and Se. freetownensis were reported only from Mauritania. Our review has updated and summarized the geographic distribution of 26 species reported so far in Morocco, Algeria, Tunisia, and Libya, excluding Mauritania from a detailed analysis due to the unavailability of accurate distribution data. In addition, morphological differences important for species identification are summarized with particular attention to closely related species such as Ph. papatasi and Ph. bergeroti, Ph. chabaudi, and Ph. riouxi, and Se. christophersi and Se. clydei.

• MeSH
• hmyz - vektory mikrobiologie   parazitologie   virologie   MeSH
• infekční nemoci epidemiologie   přenos   MeSH
• lidé MeSH
• Psychodidae mikrobiologie   parazitologie   virologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• systematický přehled MeSH
• Geografické názvy
• severní Afrika epidemiologie   MeSH

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.

• Klíčová slova
• Blood meal, Central Europe, Climate, MALDI-TOF, Phlebotomine sand fly,
• MeSH
• ekologie * MeSH
• hmyz - vektory * parazitologie   MeSH
• koně MeSH
• kur domácí MeSH
• Leishmania infantum MeSH
• Phlebotomus * genetika   MeSH
• Psychodidae MeSH
• retrospektivní studie MeSH
• roční období * MeSH
• sekvenční analýza DNA MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Evropa MeSH
• Rakousko MeSH

Leishmania major is the main causative agent of cutaneous leishmaniasis in the Old World. In Leishmania parasites, the lack of transcriptional control is mostly compensated by post-transcriptional mechanisms. Methylation of arginine is a conserved post-translational modification executed by Protein Arginine Methyltransferase (PRMTs). The genome from L. major encodes five PRMT homologs, including the cytosolic protein associated with several RNA-binding proteins, LmjPRMT7. It has been previously reported that LmjPRMT7 could impact parasite infectivity. In addition, a more recent work has clearly shown the importance of LmjPRMT7 in RNA-binding capacity and protein stability of methylation targets, demonstrating the role of this enzyme as an important epigenetic regulator of mRNA metabolism. In this study, we unveil the impact of PRMT7-mediated methylation on parasite development and virulence. Our data reveals that higher levels of LmjPRMT7 can impair parasite pathogenicity, and that deletion of this enzyme rescues the pathogenic phenotype of an attenuated strain of L. major. Interestingly, lesion formation caused by LmjPRMT7 knockout parasites is associated with an exacerbated inflammatory reaction in the tissue correlated with an excessive neutrophil recruitment. Moreover, the absence of LmjPRMT7 also impairs parasite development within the sand fly vector Phlebotomus duboscqi. Finally, a transcriptome analysis shed light onto possible genes affected by depletion of this enzyme. Taken together, this study highlights how post-transcriptional regulation can affect different aspects of the parasite biology.

• MeSH
• delece genu MeSH
• Leishmania major enzymologie   genetika   metabolismus   MeSH
• leishmanióza kožní parazitologie   patologie   MeSH
• myši MeSH
• neutrofily fyziologie   MeSH
• proteinmethyltransferasy genetika   metabolismus   MeSH
• protozoální proteiny metabolismus   MeSH
• regulace genové exprese enzymů MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Intramural MeSH
• Názvy látek
• proteinmethyltransferasy MeSH
• protozoální proteiny MeSH

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.

• Klíčová slova
• Central Europe, MALDI-TOF mass spectrometry, Transphlebotomus, autoimmunofluorescence, genotyping, leishmaniasis,
• Publikační typ
• časopisecké články MeSH