Phlebotomine Sand Flies Research Articles

Human Cutaneous Leishmaniasis in North Africa and Its Threats to Public Health: A Statistical Study Focused on Djelfa (Algeria).

Cutaneous leishmaniasis, the most common form of leishmaniasis, causes long-term skin lesions on exposed areas of the skin. It is caused by a protozoan parasite belonging to the genus Leishmania and is transmitted via infected phlebotomine sand flies. In North Africa, particularly Algeria, the disease represents a major public health problem. This retrospective study, which focuses on the agropastoral region of Djelfa (central Algeria) during a period of 16 years, from 2006 to 2021, is part of the surveillance of cutaneous leishmaniasis to identify the key factors favouring its probable spread. The analyzed data reveal that this disease is more prevalent in male patients (53.60%) and is highly widespread in this vast area of 66,415 km2 with a total of 3864 CL cases, reaching a peak of 1407 cases in 2006. Statistically, the Pearson correlation validated by the p-value shows, in an original and sometimes unexpected way, that certain factors, such as temperature linked to climate change, are playing a significant role in the probable spread of the disease in Djelfa and its surrounding regions. The concentration of the population in some specific rural areas with limited or nonexistent access to public health services is another potential factor in disease transmission. The results were highlighted by a significant correlation coefficient (r=0.66) with a p-value less than 0.01. While there is currently no vaccine or prophylactic drug available, our research represents a preliminary approach that addresses various epidemiological aspects of the disease. This paves the way for a proactive preventive strategy involving the control of vector-borne diseases.

What's Eating You? Phlebotomine Sandflies and Leishmania Parasites.

Phlebotomine sandflies of the Phlebotomus and Lutzomyia genera are vectors for the spread of Leishmania parasites. Leishmaniasis is a parasitic infection most common in the Middle East and Central and South America. Few cases have been reported in the United States, with most patients presenting after returning from travel to other countries; however, the parasite has been locally acquired in Texas and Oklahoma. Clinical presentation varies depending on the species of Leishmania. There are 4 general clinical classifications of leishmaniasis: cutaneous, diffuse cutaneous, mucocutaneous, and visceral.

First Detection and Molecular Analysis of Leishmania infantum DNA in Sand Flies of Kosovo.

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.

Phlebotomine sand flies (Diptera, Psychodidae) from Pha Tong cave, Northern Thailand with a description of two new species and taxonomical thoughts about Phlebotomus stantoni.

In South-East Asia, Thailand is the country with the highest number of human autochthonous cases of leishmaniases mostly due to Leishmania martiniquensis. Their transmission remains unresolved to date even though sand flies are known vectors of leishmaniases. As such, we focused a study on the sand fly fauna of a cave in Thailand to explore the biodiversity of potential Leishmania vectors. We carried out an inventory in Pha Tong cave. We caught and identified 570 Phlebotomine sand flies (452 females and 118 males) and identified 14 species belonging to the genera Phlebotomus, Idiophlebotomus, Chinius, Sergentomyia and Grassomyia. Among these 14 species, two could not be related to known sand fly species. Herein, we propose the description of two new sand fly species, previously unknown to science. The first new species, Phlebotomus shadenae n. sp. is a sand fly of the subgenus Anaphlebotomus. It is morphologically close to Ph. stantoni, a species widely distributed throughout Southeast Asia. However, it differs by the length of the genital filaments in males or by the length of the ducts of the spermathecae in females as well as the high divergence of cytochrome b sequences. Additionally, we revised the systematics of the subgenus Anaphlebotomus and reinstated, by examination of its holotype, the validity of Ph. maynei, an Indian wrongly considered as a synonym of Ph. stantoni in the past. The second new species, Sergentomyia maiae n. sp., differs from a species in the same group, Se. barraudi, by an original cibarial double row of vertical teeth as well as by molecular data. We propose the description of two new sand fly species for Science with morphological and molecular evidence. Ph. shadenae n. sp. was also found to be distributed in the south of Thailand and in Laos. Future studies need to determine whether these two species can play a role as vectors of Leishmania parasites, Trypanosomatids or Phlebovirus. Most of the species caught in the present study are strictly cavernicolous except Grassomyia sp. and a few Sergentomyia.

Pediatrik Kutanöz Leishmania Olgularının Değerlendirilmesi

Cutaneous Leishmaniasis (CL) is a disease caused by leishmania-type protozoans, which is transmitted by the bite of infected female phlebotomine sandflies and is characterized by ulcerated nodular lesions. Twenty-one pediatric cutaneous leishmania cases followed by pediatric infectious diseases and dermatology were included in the study. The demographic and clinical characteristics of the patients, the local or systemic treatments, and side effects were analyzed retrospectively. 14 (66%) of the patients were female and 7 (34%) were male. The mean age of the cases was 6.4 years. Fifteen of the patients were refugees (seven of the patients were from Iraq, and eight of them were from Syria). Ten of the patients ( 47%) had lesions only on the face, 6 (28%) were both on the face and hand, 4 (20%) were on the lower extremities. Seven patients (34%) had a single lesion, fourteen had multiple lesions and seven had more than four lesions. Amastigote was observed in the microbiological examination of skin scraping samples of 13 patients. Intralesional therapy was given to 15 patients, systemic treatment was given to 6 patients, and 2 patients refused systemic treatment. Five patient was given meglumine antimoniate, one patient was given amphotericin B. In one patient, side effects such as facial swelling, rash, and edema developed after amphotericin b, and the treatment was changed to meglumine antimoniate. Leishmaniasis is a chronic disease caused by flagellate protozoa of the genus Leishmania. especially in endemic countries. CL has become a relatively common condition all over the world due to international travel, migration, and refugees. Cutaneous Leishmania should be considered when there are chronic, painless skin lesions outside of endemic areas.

Survey of phlebotomine sand fly fauna in a public Zoo in Brazil: Species diversity, seasonality, and host variety.

Leishmaniasis is a dynamic disease in which transmission conditions change due to environmental and human behavioral factors. Epidemiological analyses have shown modifications in the spread profile and growing urbanization of the disease, justifying the expansion of endemic areas and increasing number of cases in dogs and humans. In the city of Belo Horizonte, located in the southeastern state of Minas Gerais (Brazil), visceral leishmaniasis (VL) is endemic, with a typical urban transmission pattern, but with different regional prevalence. This study was conducted at the Zoo of the Foundation of Municipal Parks and Zoobotany of Belo Horizonte (FPMZB-BH), located in the Pampulha region, which is among the areas most severely affected by VL. This study aimed to determine the taxonomic diversity of native phlebotomine sand flies (Diptera: Psychodidae), identify climatic variables that potentially affect the phenology of these insects, and determine the blood meal sources for female phlebotomine sand flies. To achieve this, 10 mammal enclosures in the zoo were selected using the presence of possible leishmaniasis reservoirs as a selection criterion, and sampled using light traps between August 2019 and August 2021. A total of 6034 phlebotomine sand flies were collected, indicating nine species, with Lutzomyia longipalpis being the very abundant species (65.35% of the total). Of the 108 engorged phlebotomine collected females, seven samples (6.5%) were positive for blood meals from humans, marsupials, canids, and birds. Relative humidity and rainfall increased the phenology of phlebotomine sand flies, with population increases in the hottest and wettest months. The data obtained will provide guidelines for competent health agencies to implement vector control measures to reduce the risk of leishmaniasis transmission in the FPMZB-BH.

Molecular identification of phlebotomine sand flies and the harbored Leishmania spp. in Sokoto State, Nigeria.

Female sand flies are hematophagous, feeding on animals and in the process serve as vectors for Leishmania, the parasites that cause leishmaniasis in humans. Leishmaniasis are a group of parasitic neglected tropical diseases in 98 countries including Nigeria and kills ~60,000 people/year. In Nigeria, Sokoto State is endemic to leishmaniasis but there is a knowledge gap on the identity of the prevalent sand flies and the Leishmania species they transmit. Hence, this cross-sectional study was designed to take inventory of the species of sand flies in Sokoto using genetic methods. 1,260 (310 females) sand flies were collected from three Local Government Areas (L.G.A) of Sokoto State- Wamakko, Sokoto South and Kware. Genomic DNA was extracted from each fly and DNA amplification by polymerase chain reaction (PCR) was carried out on the DNA samples using primers targeting the arthropods mitochondrial cytochrome oxidase subunit 1 (mt-coI) gene, and nested PCR with primers targeting the gene for Leishmania internal transcribed spacer-1 (its-1) of ribosomal RNA its-1rRNA. The PCR products were sequenced. Gene sequence analysis revealed five species of sand flies belonging to the old-world genera namely Phlebotomus and Sergentomyia. The identified species were P. papatasi (6.45%), S. adleri (6.45%), S. affinis (9.7%), S. distincta (9.7%), S. schwetzi (67.7%). Within the sampling period, sand flies were most abundant in the rainy months of August (104/33.5%) and September (116/37.4%) with all the five identified species occurring. Sequence analysis of its-1 gene identified Leishmania infantum in two sand flies (2/310)- P. papatasi (from Sokoto South) and S. affinis (from Wamakko). BLAST search in NCBI and phylogenetic analysis revealed that the sand fly species are related to the species reported in different parts of Africa, while the L. infantum is identical to strain reported in Brazil (KY379083.1). Phlebotomus papatasi and four species belonging to the genus Sergentomyia are the most prevalent sand flies in Sokoto State, Nigeria and they harbor L. infantum solely. The results shed light on why visceral leishmaniasis is the most predominant form of the disease. Therefore, we recommend that adequate care for dogs must be instituted as dogs are the major animal reservoir for L. infantum.

Effects of host species on microbiota composition in Phlebotomus and Lutzomyia sand flies

BackgroundBlood-sucking phlebotomine sand flies are vectors of the protozoan parasites Leishmania spp. Although the intestinal microbiota is involved in a wide range of biological and physiological processes and has the potential to alter vector competence, little is known about the factors that modify the gut microbiota composition of sand flies. As a key step toward addressing this issue, we investigated the impact of host species on the gut bacterial composition in Phlebotomus and Lutzomyia sand flies reared under the same conditions.MethodsBacterial 16S rRNA gene amplification and Illumina MiSeq sequencing were used to characterize the overall bacterial composition of three laboratory-reared sandflies: Phlebotomus papatasi, Ph. duboscqi, and Lutzomyia longipalpis.ResultsOur results showed that the larvae of the three sand fly species harbored almost the same microbes but had different relative abundances. Adult Ph. papatasi and Ph. duboscqi revealed similar microbiome compositions, which were distinct from that of adult Lu. longipalpis. Furthermore, we showed that Ph. papatasi and Ph. duboscqi are hosts for different bacterial genera. The experiment was repeated twice to improve accuracy and increase reliability of the data, and the same results were obtained even when a distinct composition of the microbiome among the same species was identified probably because of the use of different larvae food batch.ConclusionsThe present study provides key insights into the role of host species in the gut microbial content of different sand fly species reared under the same conditions, which may influence their susceptibility to Leishmania infection.Graphical

Species diversity and detection of pathogens in phlebotomine sand flies collected from forest management areas of Quintana Roo, Mexico.

Sand flies have expanded their areas of distribution, thereby increasing the risk of pathogen transmission in non-endemic areas. To establish efficient prevention and control strategies for the transmission of vector-borne pathogens, it is important to understand seasonal dynamics of their vectors. In Mexico, there are several areas where the contact between sand flies, hosts and reservoirs favours the transmission of the pathogen. We compared sand fly communities in a forest management area and a conserved area in Noh-Bec, Quintana Roo, Mexico. The analysis included species diversity, activity peaks and molecular detection of pathogens. Sand flies were collected from November to December 2021 and April to May 2022, during 84 night-traps. The conserved area showed higher numbers and greater species heterogeneity of sand flies as compared with the other sites. The β-diversity analysis revealed that sites disturbed by logging (S1, S2, S3) had greater similarity (90%) in their sand fly species composition than a conserved area (S4) (similarity = 36%). Although none of the specimens were infected with Leishmania, we detected Wolbachia (19.4%) in all four sites, as well as Bartonella (3.25%) only in the disturbed sites. Further studies on the dynamics of sand fly populations and their association with pathogens are necessary.

The Phlebotomine sand fly fauna of Switzerland revisited.

Sand flies (Diptera: Psychodidae, Phlebotominae; Newstead, 1911) are widespread in Europe, being particularly common in the Mediterranean region but rare north of the Alps. Thus, Switzerland is an opportune place to investigate the sand fly fauna on both sides of the Alpine crest, in southern sub-Mediterranean climate and northern oceanic temperate climate. We reinvestigated the Swiss sand fly fauna with the aim to assess changes in composition, altitudinal distribution, abundance and seasonality. Thirty-eight sites were investigated with light traps and/or interception sticky traps in 4 years. Ninety and 380 specimens were caught by light traps and sticky traps, respectively, at 15 collecting sites. Four species were identified. Phlebotomus mascittii (Grassi, 1908), Phlebotomus perniciosus (Newstead, 1911) and Sergentomyia minuta (Rondani, 1843) were confirmed in Ticino, and P. mascittii for the first time in neighbouring Grisons. Also, Phlebotomus neglectus(Tonnoir, 1921) is for the first time reported, though at a very low density compared to P. perniciosus at the same site. Its presence in Ticino supports the northward spread observed in Italy. Sand flies were detected north of the Alps at one site only, endorsing a historical report. Overall, the low density of P. perniciosus and very low density of P. neglectus suggest that canine leishmaniosis may not be an important disease risk in Switzerland.

Diversity and spatio-temporal variation of phlebotomine sand flies (Phlebotominae: Diptera: Psychodidae) in three different types of land use and seasons in the state of Veracruz, Mexico

A sand fly fauna inventory was conducted in an ecotourism area of central Veracruz, Mexico. We recorded and analyzed the sand fly diversity at 3 different land use types (well-preserved forest, fruit-tree plantations, and human settlements) with sampling conducted 3 times in 1 year (dry, rainy, and cold seasons). A total of 891 specimens of 6 genera and 14 species was recorded. High diversity was detected in the preserved area due to a high species richness and abundance as compared with the fruit-tree plantations and human settlements, respectively. In relation to the seasons of the year, high diversity was found in the cold season as compared with the rainy and dry seasons. The variation of the Phlebotominae assemblage in space and time in the ecotourism zone is described, serving as a baseline to recommend preventive actions to settlers and travelers. Additionally, we include new species records for the state of Veracruz.

Phlebovirus detection on phlebotomine sandflies in Lampedusa Island (Italy).

Phleboviruses are common human pathogens diffused on the Mediterranean area whose infection can cause the typical prodromal symptom of a mild three‑days fever. In particular, Toscana Virus (TOSV) has a great concern since its capacity to provoke central nervous system disorders like meningoencephalitis. Furthermore, as the phlebotomine arthropod vectors represent the main carrier for pathogens of the genus Leishmania as well, the purpose of the study was to investigate the presence of TOSV in Lampedusa, Italy previously reported for leishmaniosis infection cases. The survey was carried out through an initial sampling phase of sand flies, by means of CDC light traps, and a second step of molecular analyses. The genomic S‑segment of TOSV was targeted. The positive samples were sequenced and compared with those available in GenBank™ using Basic Local Alignments Tool (BLAST) analyses. The study revealed for the first time the presence of TOSV in Lampedusa, Italy. The entomological studies directed on vectors are currently widely used in sand fly surveillance, and new data on TOSV are of public health concern.

Morphological and DNA-based description of Trichophoromyia peixotoi n. sp. (Diptera: Psychodidae), a new sand fly species from the Brazilian Amazon

Background Phlebotomine sand flies of the genus Trichophoromyia Barretto, 1962 are of great relevance to public health as vectors of Leishmania protozoans. A new phlebotomine species named Trichophoromyia peixotoi n. sp. is here described based on both male morphology and COI DNA barcodes.MethodsThe sand fly specimens were collected in the Parque Nacional da Amazônia (PNA), situated in the municipality of Itaituba, state of Pará, Brazil. Morphological description was done based on 10 male specimens. Five specimens were DNA barcoded for the COI gene.ResultsThe morphological and molecular analyses allowed the delimitation of this new species from others of Trichophoromyia. Trichophoromyia peixotoi n. sp. is closely related to other species with aedeagal ducts > 4 times the length of the sperm pump, from which it may be distinguished by the gonocoxite bristles and paramere shape.ConclusionsThe description of T. peixotoi n. sp. brings the number of species of Trichophoromyia to 45, including 24 for Brazil. The integrative taxonomy effort through the analysis of COI barcodes proved to be effective in the species delimitation of some Trichophoromyia spp.Graphical

Leishmania Seroprevalence in Dogs: Comparing Shelter and Domestic Communities.

Canine leishmaniosis (CanL) is a chronic, systemic, and often severe disease. The main causative agent of CanL is a protozoan parasite, Leishmania infantum, with phlebotomine sand flies acting as vectors. In Europe and other continents, L. infantum is also responsible for leishmaniosis in other animals, such as cats, horses, and humans. In Portugal, animal and human leishmaniosis is endemic, and high prevalence levels of infections and disease have been reported in dogs. There is a prejudice against stray animals and also those housed in shelters, assuming they have higher levels of infection with vector-borne pathogens, including L. infantum, when compared to domestic animals. In northeastern Portugal, serum samples were obtained from March to May 2022 in three shelters (n = 179) and thirteen veterinary clinics (n = 164), resulting in 343 dogs being analyzed for antibodies to Leishmania spp. by the direct agglutination test (DAT). The overall seroprevalence was 9.9%, with 15.2% seroprevalence in domestic dogs and 5.0% in the shelter ones (p = 0.003). The fact that shelter dogs had a lower seroprevalence could be explained by more regular veterinary care provided in shelters regarding preventive measures, including insecticides with an antifeeding effect, in comparison with domestic dogs.

Blood meal analysis and molecular detection of mammalian Leishmania DNA in wild-caught Sergentomyia spp. from Tunisia and Saudi Arabia.

Phlebotomine sand flies (Diptera: Phlebotominae) belonging to the genus Phlebotomus are vectors of pathogens such as arboviruses, bacteria, and parasites (Leishmania). Species of the genus Sergentomyia (Se.) transmit Sauroleishmania (Reptile Leishmania) and feed on cold-blooded vertebrates; recently, they have been incriminated in mammalian Leishmania transmission. In addition, they have been reported to feed on warm-blooded vertebrates. This study aimed to (i) screen wild-caught Sergentomyia species for the detection of mammalian Leishmania and (ii) identify the blood meal origin of engorged females. The sand flies were collected using centers for disease control and prevention (CDC) traps, mounted and identified morphologically. Only females of the genus Sergentomyia were screened for Leishmania infection using PCR targeting the 18S ribosomal DNA locus. For positive specimens, Leishmania parasites were typed using nested PCR targeting ribosomal internal transcribed spacer 1 followed by digestion with HaeIII. The PCR-RFLP results were confirmed through sequencing. Blood meal identification was performed through PCR amplification of the vertebrate cytochrome b gene using degenerate primers followed by sequencing. In total, 6026 sand fly specimens were collected between 2009 and 2018. Among these, 511 belonged to five species of Sergentomyia genus: Se. minuta (58.51%), Se. fallax (18.01%), Se. clydei (14.68%), Se. dreyfussi (6.26%), and Se. antennata (2.54%). A total of 256 female Sergentomyia sp. specimens were screened for Leishmania infection. Seventeen (17) were positive (6.64%). Two Leishmania species were identified. Leishmania major DNA was detected in five specimens; this included three Se. fallax, one Se. minuta, and one Se. dreyfussi collected from Tunisia. Leishmania infantum/L. donovani complex was detected in four Se. minuta and three Se. dreyfussi specimens collected from Tunisia. In addition, we identified the blood meal origin of five engorged Se. minuta specimens collected from Tunisia. Sequencing results revealed two blood sources: humans (n = 4) and reptiles (n = 1) indicating possible role of Sergentomyia species in the transmission of human Leishmania. In addition, these species could be involved in the life cycle of L. infantum/L. donovani complex and L. major. The results of the blood meal origin showed that Sergentomyia fed on both cold- and warm-blooded vertebrates. These findings enable a better understanding of the behavior of this sand fly genus. Further studies should focus on the role of Sergentomyia in human Leishmania transmission and possible control of this disease.

Occurrence of Phlebotomus mascittii (Diptera: Psychodidae: Phlebotominae) in the Upper Rhine Valley of Alsace, France

Summary Because of the apparent northwards spread of canine leishmaniosis cases, we decided to investigate the occurrence of the potential vector species, namely Phlebotomine sand flies, in the Upper Rhine valley of Alsace, France. Trappings were performed in the natural regions of the Vosges foothills and the Alsatian Jura, which looked the most suitable for sand flies, considering climate and environment. Fifteen locations (25 sites) were investigated by means of light traps and/or interception sticky traps in 2009, 2011 and 2018. Phlebotomus mascittii Grassi, 1908 was the only sand fly species identified, and it was detected at three locations of the Vosges foothills, in both 2011 and 2018. This study confirms the occurrence of P. mascittii in the Upper Rhine valley of Alsace, around 70 years after the first and single historical report. Our data do not suggest the recent northwards spread of the species. The observed low density of Phlebotominae suggests that the risk for Leishmania transmission in Alsace, if introduced, is very low.

Phylogenetic relations among Mexican phlebotomine sand flies (Diptera: Psychodidae) and their divergence time estimation.

Phlebotomine sand flies (Diptera: Psychodidae: Phlebotominae) have biological relevance as vectors of several pathogens. To ensure periodic entomological monitoring it is necessary to have efficient and accurate tools for an adequate taxonomic identification. There are only few studies on phylogenetic analyses of phlebotomine sand flies from Neotropics, based mostly on morphological and/or molecular data, which makes the delimitation of intra- and interspecific variability of species challenging. Here we generated new molecular information on sand fly species distributed in endemic areas of leishmaniasis in Mexico, using mitochondrial and ribosomal genes, and incorporating morphological information available. Specifically, we established their phylogenetic relationships, and estimated their divergence time. Our study provides molecular information for 15 phlebotomine sand fly species from different areas of Mexico, contributing to the genetic inventory and phylogenetic relations among Neotropical species of the subfamily Phlebotominae. Mitochondrial genes proved to be suitable markers for the molecular identification of phlebotomine sand flies. However, the incorporation of additional nuclear gene information could increase the significance of phylogenetic inferences. We also provided evidence about a possible divergence time of phlebotomine sand fly species, supporting their presumable origin in the Cretaceous period.

First evidence of experimental genetic hybridization between cutaneous and visceral strains of Leishmania donovani within its natural vector Phlebotomus argentipes

Leishmaniasis is a neglected tropical disease caused by protozoan parasites of genus Leishmania, and transmitted by different species of Phlebotomine sand flies. More than 20 species of Leishmania are known to cause disease in humans and other animals. Leishmania donovani species complex is known to have a vast diversity of clinical manifestations in humans, but underlying mechanisms for such diversity are yet unknown. Long believed to be strictly asexual, Leishmania have been shown to undergo a cryptic sexual cycle inside its sandfly vector. Natural populations of hybrid parasites have been associated with the rise of atypical clinical outcomes in the Indian subcontinent (ISC). However, formal demonstration of genetic crossing in the major endemic sandfly species in the ISC remain unexplored. Here, we investigated the ability of two distinct variants of L. donovani associated with strikingly different forms of the disease to undergo genetic exchange inside its natural vector, Phlebotomus argentipes. Clinical isolates of L. donovani either from a Sri Lankan cutaneous leishmaniasis (CL) patient or an Indian visceral leishmaniasis (VL) patient were genetically engineered to express different fluorescent proteins and drug-resistance markers and subsequently used as parental strains in experimental sandfly co-infection. After 8 days of infection, sand flies were dissected and midgut promastigotes were transferred into double drug-selective media. Two double drug-resistant, dual fluorescent hybrid cell lines were recovered, which after cloning and whole genome sequencing, were shown to be full genomic hybrids. This study provides the first evidence of L. donovani hybridization within its natural vector Ph. argentipes.

Molecular Detection of Leishmania in Wild Caught Sand Flies of Larroussius Subgenus in Iran: Combined Use of Internal and External Morphological Characters as a Mean to Differentiate Morphologically Similar Females.

Phlebotomine sand flies are the primary vectors of leishmaniasis; visceral form of disease is transmitted mainly by species belonging to Larroussius and Adlerius subgenus. Species identification of some females in Larroussius subgenus is not easy due to the high similarity. Accurate species identification enables control operations to target only main vectors and increase understanding of ecological needs, bionomic characteristics, and behavioral traits. The purpose of current study was to use two approaches based on internal and external morphological characters to identify the wild caught of female specimens belonged to Larroussius subgenus and investigate Leishmania infection. Totally, 128 specimens belonging to Larroussius' subgenus were collected from a VL focus in northwestern Iran, species differentiation was done by two approaches proposed in literature: (1) characters of pharyngeal armature, number of Spermathecal segment, length of spermathecal neck, palpal and ascoid formulae; (2) blindly on the basis of the shape of the base of spermathecal duct. Their potential infection to Leishmania was investigated by kDNA-Nested-PCR. The results of species identification were consistent based on two methods. Among three identified species, Phlebotomus perfiliewi was found to be the dominant species, followed by Ph. neglectus and Ph. tobbi. Two specimens of Ph. perfiliewi were found to be infected by Leishmania infantum, which emphasizes the role of this species in VL transmission in study area. It is suggested that combination of the characters used here be considered for species identification of female of Larroussius subgenus to take advantage of maximum characters, especially where species present sympatrically.

Identification of molecular mechanisms causing skin lesions of cutaneous leishmaniasis using weighted gene coexpression network analysis (WGCNA)

Leishmaniasis is an infectious disease, caused by a protozoan parasite. Its most common form is cutaneous leishmaniasis, which leaves scars on exposed body parts from bites by infected female phlebotomine sandflies. Approximately 50% of cases of cutaneous leishmaniasis fail to respond to standard treatments, creating slow-healing wounds which cause permanent scars on the skin. We performed a joint bioinformatics analysis to identify differentially expressed genes (DEGs) in healthy skin biopsies and Leishmania cutaneous wounds. DEGs and WGCNA modules were analyzed based on the Gene Ontology function, and the Cytoscape software. Among almost 16,600 genes that had significant expression changes on the skin surrounding Leishmania wounds, WGCNA determined that one of the modules, with 456 genes, has the strongest correlation with the size of the wounds. Functional enrichment analysis indicated that this module includes three gene groups with significant expression changes. These produce tissue-damaging cytokines or disrupt the production and activation of collagen, fibrin proteins, and the extracellular matrix, causing skin wounds or preventing them from healing. The hub genes of these groups are OAS1, SERPINH1, and FBLN1 respectively. This information can provide new ways to deal with unwanted and harmful effects of cutaneous leishmaniasis.