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Background: Phlebotomine sand flies (Diptera: Psychodidae) are important vectors of various pathogens, mainly Leishmania parasites. In the Old World, the most important genus in term of pathogens transmission is the genus Phlebotomus, which includes many proven or suspected vectors of several Leishmania species, while the genus Sergentomyia remains so far unproven as a vector of human pathogens. Algeria is one of the most affected countries by human leishmaniasis. Methods: In the present study, an entomological survey was carried out in two provinces, Ghardaïa and Illizi, located in the north and central Sahara, respectively, where cases of human leishmaniasis are recorded. Our goal was to understand the role of the local sand fly species in the transmission of Leishmania parasites and to analyze their blood meal preferences. Collected sand flies were identified by a combination of morphological and molecular approaches that included DNA-barcoding and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) protein profiling. In addition, female blood meals were analyzed by peptide mass mapping using MALDI-TOF MS. Results: In total, 640 sand fly specimens belonging to Phlebotomus and Sergentomyia genera were collected in the two provinces. Sergentomyia antennata and Se. fallax were most abundant species in Ghardaïa, and Ph. papatasi and Ph. alexandri in Illizi. In addition, a new sand fly species was described in Illizi named Sergentomyia (Sergentomyia) imihra n. sp. Blood meal analysis of the engorged females revealed various mammalian hosts, especially goats, but also humans for Phlebotomus papatasi and Ph. alexandri, suggesting that these vector species are opportunistic feeders. Conclusions: Integrative approach that combined morphological analysis, sequencing of DNA markers, and protein profiling enabled the recognition and description of a new Sergentomyia species, raising the number of the Algerian sand fly fauna to 27 species. Further sand fly surveillance in the central Sahara is recommended to identify the thus-far unknown males of Se. imihra n. sp. [ABSTRACT FROM AUTHOR]

Background: Sand flies, belonging to the Psychodidae family, represent small, hairy insects that serve as significant vectors in various important medical and veterinary diseases. Despite being recognized by the World Health Organization as an endemic area for leishmaniasis, Southeast Asia lacks comprehensive information on the species composition and biology of sand flies. To address this, the current study aimed to survey sand fly biodiversity. Methods: Sand flies from six provinces in Southern Vietnam were collected using CDC light traps. Sand flies were subsequently identified morphologically and confirmed molecularly using mitochondrial cytochrome oxidase c subunit I (COI) and cytochrome b (cytb) sequences. BLASTN searches were conducted, and the species identity of sand flies was further confirmed through a Barcode of Life Database (BOLD) search utilizing COI sequences. Subsequently, nucleotide sequences were subjected to a panel of analyses including intraspecific variation, phylogenetic relationships and haplotype network. The average densities of collected sand flies (sand flies/trap/night) and species richness were also recorded. Results: A total of 753 sand flies were collected. After excluding damaged specimens, six sand fly species, namely Phlebotomus stantoni, Sergentomyia khawi, Se. silvatica, Se. barraudi, Se. bailyi and Grassomyia indica, were identified. All conspecific sand fly sequences, including Ph. stantoni, Se. barraudi, Gr. indica, Se. bailyi, Se. khawi and Se. silvatica, clustered with their reference sequences, corroborating the results of morphology-based identification, BLASTN analysis and BOLD search. For intraspecific variation of sand flies obtained from the current study, COI diversity indices were consistently higher than those of cytb. Conclusions: This study provides the first updates on morphological and molecular characterization of sand flies in Southern Vietnam. This acquired knowledge on sand fly species composition is essential for controlling sand fly-borne diseases in this potentially endemic region. [ABSTRACT FROM AUTHOR]

Background: Several new species of Leishmania have recently emerged in Europe, probably as the result of global changes and increased human migration from endemic areas. In this study, we tested whether two sand fly species, the Western Mediterranean Phlebotomus perniciosus and the Eastern Mediterranean P. tobbi, are competent vectors of L. donovani, L. major and L. martiniquensis. Methodology/principal findings: Sand flies were infected through the chick skin membrane using Leishmania species and strains of various geographical origins. Leishmania infections were evaluated by light microscopy and qPCR, and the representation of morphological forms was assessed from Giemsa-stained gut smears. Neither P. perniciosus nor P. tobbi supported the development of L. martiniquensis, but L. major and L. donovani in both species survived defecation of blood meal remnants, colonized the stomodeal valve and produced metacyclic stages. The results with L donovani have shown that infection rates in sand flies can be strain-specific; therefore, to determine vector competence or refractoriness, it is optimal to test at least two strains of Leishmania. Conclusions, significance: Both sand fly species tested are potential vectors of L. donovani and L. major in Mediterranean area. However, further studies will be needed to identify European vectors of L. martiniquensis and to test the ability of other European sand fly species to transmit L. major, L. donovani, L. tropica and L. infantum. Author summary: Leishmaniases are serious human and animal diseases caused by protozoa of the genus Leishmania (Kinetoplastida: Trypanosomatidae), transmitted by blood-feeding insects–phlebotomine sand flies (Diptera: Psychodidae). They are mainly distributed in tropical and subtropical regions, but recently the ranges of the vectors have spread more into temperate regions. In Europe, species of Leishmania that were previously absent have emerged, mainly due to the mass migration of the refugees from endemic areas. It is therefore important to know whether local sand flies can transmit these "new" Leishmania species. In our study, we tested two widely distributed sand fly species, P. perniciosus and P. tobbi. In both of them, L. major and L. donovani completed their development, thus P. perniciosus and P. tobbi are potential vectors of these parasites. The third Leishmania species tested, L. martiniquensis, which has been repeatedly recorded in central Europe, did not survive in P. perniciosus and P. tobbi; its vectors have yet to be confirmed. [ABSTRACT FROM AUTHOR]

Background: Western Ghats is a biodiversity treasure trove with reports of indigenous leishmaniasis cases. Hence, systematic sand fly surveillance was carried out among the tribal population. The present study reports a novel sand fly species, Phlebotomus (Anaphlebotomus) ajithii n. sp. (Diptera: Psychodidae), discovered in the Western Ghats of India. Methods: A comprehensive sand fly survey was conducted across the Kollam, Thrissur, Idukki, Kasaragod and Malappuram districts of Kerala, India. The survey spanned both indoor and outdoor habitats using standard collection methods over a 3-year, 3-month period. DNA barcoding of samples was performed targeting mitochondrial cytochrome c oxidase subunit I (COI) gene, and the sequence generated was subjected to phylogenetic analysis. Results: Phlebotomus (Anaphlebotomus) ajithii, a new sand fly species, is recorded and described in this communication. The morphological relationship of the new species to other members of the subgenus Anaphlebotomus is discussed. Mitochondrial COI barcode followed by phylogenetic analysis confirmed that specimens of Ph. ajithii belong to the same taxonomic group, while a genetic distance of 11.7% from congeners established it as a distinct species. Conclusions: The Western Ghats, known for its rich biodiversity, has lacked systematic entomological surveys focusing on sand flies. This study aims to fill this gap and reports and describes a new species of sand fly. [ABSTRACT FROM AUTHOR]

Background: Southeast Asia is well known as a hotspot of biodiversity. However, very little is known about cave-dwelling hematophagous insects that are medically important. Taxonomic knowledge and ecology of phlebotomine sand flies are very poorly studied in Laos, as well as in other countries in the region. Herein, we report species diversity data and some notes on the ecology of the detected species from these karstic limestone areas of Laos. Methods: Phlebotomine sand flies were collected using Centers for Disease Control and Prevention (CDC) light traps from limestone cave locations in three districts of Vientiane Province, Laos. Both morphological and molecular techniques were used for sand fly identification. Species diversity and abundance were analyzed according to sites, locations, collection seasons, and trapping positions. Results: A total of 6564 sand flies, of which 5038 were females and 1526 were males, were morphologically identified into 20 species belonging to five genera (Chinius, Idiophlebotomus, Phlebotomus, Sergentomyia, and Grassomyia). The most abundant species were Chinius eunicegalatiae, Phlebotomus stantoni, Sergentomyia hivernus, Se. siamensis, and Idiophlebotomus longiforceps. Cytochrome b analysis results supported the morphological identification and revealed that Se. siamensis was separated from other members of the Se. barraudi group. Two new species, Se. dvoraki n. sp. and Se. marolii n. sp., were described. Sand fly density was generally high except in a cave in Vangvieng, with species richness ranging from 14 to 18 across different caves. Outside caves had higher species richness (R = 20) and diversity (H = 2.50) than cave entrances (R = 18, H = 2.41) and interiors (R = 16, H = 2.13). Seasonal variations showed high sand fly density in Feung and Hinheup during both dry and rainy seasons, while Vangvieng had a notable decrease in density during the dry season (D = 6.29). Conclusions: This study revealed that the diversity of phlebotomine sand fly fauna in Laos, particularly in karstic limestone areas, is greater than previously known. However, the taxonomic status of many species in Laos, as well as Southeast Asia, still needs more in-depth study using both morphological characters and molecular methods. Many species could be found from inside, at the entrance, and outside of caves, indicating a wide range of host-seeking behavior or possible natural breeding in the karstic cave areas. [ABSTRACT FROM AUTHOR]

Introduction. Pintomyia (Pifanomyia) longiflocosa is an endemic species from Colombia, found between the central and eastern Andes, and reported as one of the primary vectors of cutaneous leishmaniasis in coffee-growing zones of the country. This species is classified in the Townsendi series and can only be identified by the morphology of the male adults. Objective. To determine the potential use of the fourth larval stage of the vector Pi. longiflocosa in morphological taxonomy based on the description of its chaetotaxy. Materials and methods. Pintomyia longiflocosa adults were captured in Campoalegre, Huila, and reared in the Entomology Laboratory at the Colombian Instituto Nacional de Salud. To identify the setae found in each corporal segment, 15 fourth-instar larvae were mounted on microscope slides using Canadian balm after being cleared with 10 % potassium hydroxide and saturated phenol. Additionally, five specimens were prepared for their observation by scanning electron microscopy. Results. Based on the description of Pi. longiflocosa, we established that all species of the subgenus Pifanomyia so far described have the same antennal morphology and clavate setae along their body. However, various setae present in Pi. longiflocosa are absent in Pi. youngi, suggesting differences among the larvae of the Townsendi series. Conclusions. These results support the potential importance of morphological characters from the fourth larval instar, such as antennal morphology and chaetotaxy, specifically in closely related species that are cryptic in their adult stages.

In this study, we identified and assembled a strain of American nodavirus (ANV) in the Phlebotomus papatasi‐derived PP9ad cell line. This strain most closely resembles Flock House virus and ANV identified in the Drosophila melanogaster S2/S2R cell line. Through small RNA sequencing and analysis, we demonstrate that ANV replication in PP9ad cells is primarily targeted by the exogenous small interfering RNA (exo‐siRNA) pathway, with minimal engagement from the PIWI‐interacting RNA (piRNA) pathway. In mosquitoes such as Aedes and Culex, the PIWI pathway is expanded and specialised, which actively limits virus replication. This is unlike in Drosophila spp., where the piRNA pathway does not restrict viral replication. In Lutzomyia sandflies (family Psychodidae), close relatives of Phlebotomus species and Drosophila, there appears to be an absence of virus‐derived piRNAs. To investigate whether this absence is due to a lack of PIWI pathway proteins, we analysed the piRNA and siRNA diversity and repertoire in PP9ad cells. Previous assemblies of P. papatasi genome (Ppap_1.0) have revealed a patchy repertoire of the siRNA and piRNA pathways. Our analysis of the updated P. papatasi genome (Ppap_2.1) has shown no PIWI protein expansion in sandflies. We found that both siRNA and piRNA pathways are transcriptionally active in PP9ad cells, with genomic mapping of small RNAs generating typical piRNA signatures. Our results suggest that the piRNA pathway may not respond to virus replication in these cells, but an antiviral response is mounted via the exo‐siRNA pathway. [ABSTRACT FROM AUTHOR]

Sand flies (Diptera: Psychodidae: Phlebotominae) are blood‐feeding insects that transmit the protozoan parasites Leishmania spp. and various arboviruses. The Balkan region, including the Republic of Kosovo, harbours a diverse sand fly fauna. Vector species of Leishmania infantum as well as phleboviruses are endemic; however, recent data are scarce. We performed a cross‐sectional study to update the current sand fly distribution in Kosovo and assess biological as well as environmental factors associated with sand fly presence. CDC light trapping was conducted at 46 locations in 2022 and 2023, specifically targeting understudied regions in Kosovo. Individual morphological species identification was supported by molecular barcoding. The occurrence data of sand flies was used to create distribution maps and perform environmental analyses, taking elevation, wind speed and climate‐related factors into account. In addition, PCR‐based blood meal analysis and pathogen screening were conducted. Overall, 303 specimens of six sand fly species were trapped, predominated by Phlebotomus neglectus (97%). Barcodes from eight of nine known endemic sand fly species were obtained. Combining our data with previous surveys, we mapped the currently known sand fly distribution based on more than 4000 specimens at 177 data points, identifying Ph. neglectus and Ph. perfiliewi as the predominant species. Environmental analyses depicted two geographical groups of sand flies in Kosovo, with notable differences between the species. In total, 223 blood meals of five sand fly species were analysed. Of seven identified host species, the predominant blood meal source was observed to be cattle, but the DNA of dogs and humans, among others, was also detected. This study assessed biological as well as ecological factors of sand fly occurrence, which should help better understand and evaluate potential hot spots of disease transmission in Kosovo. [ABSTRACT FROM AUTHOR]

Copyright of Biomédica: Revista del Instituto Nacional de Salud is the property of Instituto Nacional de Salud of Colombia and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Our study aimed to develop an optimised laboratory protocol ensuring the preservation of morphological structures and extraction of high-quality DNA sequences from Psychodidae (Insecta, Diptera) specimens. With 310 analysed specimens, we investigated the impact of distinct laboratory treatments by employing two shaking categories (constant and interrupted) with five different incubation periods (16, 12, 8, 4 and 2 hours) during the DNA extraction process. Notably, 80.65% of the specimens exhibited morphological changes during DNA extraction. Our results indicated no statistical difference between constant and interrupted shaking for the total of morphological structures lost. However, within each shaking category, the loss of structures was influenced significantly by the incubation period. Prolonged incubation correlated with increased structural losses, whereas shorter incubation periods caused minor alterations in structures lost. In addition, our results showed a significant difference between constant and interrupted shaking treatments for DNA concentration. Likewise, the incubation period showed differences within each shaking category. Successful COI sequencing was achieved in 89.6% of specimens, with negligible differences in DNA fragment lengths across treatments. Our findings underscore the importance of an optimised protocol and its potential in systematic research involving nematoceran dipteran specimens by balancing morphological integrity and DNA extraction efficiency.

Background: Pintomyia longiflocosa is considered a vector of etiological agents of cutaneous leishmaniasis. The objective of this research was to establish the parameters of the life cycle of Pi. longiflocosa in semi-controlled conditions in a rural area of the Campoalegre municipality, Huila, Colombia. Methodology/principal findings: The life cycle of individuals of Pi. longiflocosa, obtained from two cohorts of collected, fed, and individualized females, was monitored during two different times of the year (between February and August 2020 and between July 2020 and January 2021, respectively). Determining parameters associated with the fertility and fecundity, time and attributes of development and survival, and its association with abiotic variables. The average duration of Cycle 1 (C1) and Cycle 2 (C2) was 134.9 and 148.78 days, respectively. The gonotrophic cycle of parental females presented significant differences (p-value <0.05) between C1 and C2 (8.47 and 11.42 days) as well as between fecundity and fertility parameters. The number of days it takes the development of the immature stages between the two cycles studied, also showed significant differences in the larvae II (15.21 and 22.23), larvae III (11.93 and 17.56), and pupae (24.48 and 22.9) stages. During C1, the survival rate was higher and consistent with the productivity of adult individuals (F1), compared to C2. Fecundity and fertility values were significantly higher in C2. Finally, a significant correlation between the number of individuals and temperature was evidenced in C1 while, for C2, there was a negative correlation with precipitation. Conclusions/significance: Significant differences were found in several biological and reproductive parameters between the two cycles monitored. The parameters of the life cycle of Pi. longiflocosa in its natural habitat would be influenced by environmental factors related to the annual seasonality in the sub-Andean rural area, conditioning the temporal distribution of this species and, consequently, the possible transmission of causative agents of cutaneous leishmaniasis. Author summary: Cutaneous leishmaniasis (CL) is a public health disease that affects more than 300 million people in tropical areas in the world and its causative agent is a parasite of the genus Leishmania. In recent decades, the cases in Latin America have increased, corresponding to 6% of cases in the tropical regions of the planet. An insect potential vector associated with the transmission of CL in these tropical and endemic areas is Pintomyia longiflocosa. Therefore, it is very important to investigate the environmental parameters that can influence its life cycle and its vector competence and vector capacity, aspects that are not fully known. In Colombia, an endemic focus is located in the Huila state, in a rural area of the municipality of Campoalegre, where this study was carried out. The authors found the relationship between some environmental parameters with the life cycle of Pi. longiflocosa under semi-controlled conditions in forested habitats, aspects that were not known until now. The impact of temperature, relative humidity, and precipitation on the gonotrophic cycle, fecundity, fertility, time of development and survival, was evidenced. This research provides information on the environmental conditions that directly affect the possible transmission of the agents that cause cutaneous leishmaniasis. [ABSTRACT FROM AUTHOR]

Background: Arthropods vector a multitude of human disease-causing organisms, and their geographic ranges are shifting rapidly in response to changing climatic conditions. This is, in turn, altering the landscape of disease risk for human populations that are brought into novel contact with the vectors and the diseases they carry. Sand flies in the genera Lutzomyia and Pintomyia are vectors of serious disease-causing agents such as Leishmania (the etiological agent of leishmaniasis) and may be expanding their range in the face of climate change. Understanding the climatic conditions that vector species both tolerate physiologically and prefer behaviorally is critical to predicting the direction and magnitude of range expansions and the resulting impacts on human health. Temperature and humidity are key factors that determine the geographic extent of many arthropods, including vector species. Methods: We characterized the habitat of two species of sand flies, Lutzomyia longipalpis and Pintomyia evansi. Additionally, we studied two behavioral factors of thermal fitness–thermal and humidity preference in two species of sand flies alongside a key aspect of physiological tolerance–desiccation resistance. Results: We found that Lu. longipalpis is found at cooler and drier conditions than Pi. evansi. Our results also show significant interspecific differences in both behavioral traits, with Pi. evansi preferring warmer, more humid conditions than Lu. longipalpis. Finally, we found that Lu. longipalpis shows greater tolerance to extreme low humidity, and that this is especially pronounced in males of the species. Conclusions: Taken together, our results suggest that temperature and humidity conditions are key aspects of the climatic niche of Lutzomyia and Pintomyia sand flies and underscore the value of integrative studies of climatic tolerance and preference in vector biology. [ABSTRACT FROM AUTHOR]

We tested the hypothesis that bioclimatic conditions affect the species richness, abundance, and diversity of sand flies in neighboring physiographic regions with endemic transmission of cutaneous leishmaniasis. A sampling program was performed in southeastern Mexico in the High Plateau of Chiapas and the Eastern Mountains physiographic regions, between April 2009 and March 2011. CDC light traps were placed in five randomly selected houses, four CDC traps in four transects in the surrounding vegetation zones, and a modified Magoon trap baited with one protected person was installed at the periphery of the villages at each study site. A total of 12,349 sand flies were identified as species, of which 98.2% were collected in the Eastern Mountains and the rest in the High Plateau of Chiapas. The Eastern Mountains region recorded a significantly higher diversity (H' = 2.04), with a species richness of 27 sand flies species and a higher evenness index (J´ = 0.62) compared to the High Plateau of Chiapas region (H' = 1.02, S = 10, J´ = 0.31). Psychodopygus panamensis (Shannon) (Diptera: Psychodidae) and Pintomyia (Pifanomyia) ovallesi (Ortiz) were the most abundant species in the Eastern Mountains, whereas Lutzomyia (Tricholateralis) cruciata (Coquillett) was most abundant in the High Plateau of Chiapas. We conclude that the differences in species richness, diversity, and abundance of sand flies between these two physiographic regions are due to the bioclimatic effects that in the Eastern Mountains are favorable for the formation of microhabitats due to the warm-warm conditions and the humid-temperate conditions which act as limiting factors in the High Plateau of Chiapas region. In addition, changes in land use due to anthropogenic activities significantly affected the structure of sand fly communities in both regions. [ABSTRACT FROM AUTHOR]

Sandfly, Phlebotomus papatasi (Diptera: Psychodidae) is the potential vector of leishmania in Egypt. In this study, an attempt was made to investigate the ultrastructure and histopathological alteration in the midgut of Phlebotomus papatasi post-treatment with methoprene (IGR) and B. sphaericus 6184 and 6446 using transmission electron micrographs (TEM). After 48 h, the TEM of the midgut treated with methoprene (IGR) showed sloughing of the periepithelial layer, and overgrowth of the brush border of the midgut wall. After 48 h post-treatment, B. sphaericus 6184 spores were discovered in the lumen and the bacterium began to enter the microvilli. Certain ultrastructural abnormalities were found in the midgut of treated P. papatasi larvae after 48 h of treatment with B. sphaericus 6446, the cytoplasm was distinguished by many vacuoles and a broken brush border. Using the light microscope, IGR-treated third instar larvae showed brush boundary overgrowth and multifocal disintegration of the midgut wall. Focused areas were sloughed and epithelial cells were dissociated from each other in larvae treated with B. sphaericus 6184. The 3rd instar P. papatasi larvae treated with B. sphaericus 6446 showed clear alterations such as total epithelial cell degradation. These results demonstrated that B. sphaericus 6446 showed the best results and great effect on P. papatasi 3rd instar larvae. More research is needed to establish the appropriate control strategies for the many leishmaniasis vectors in their various ecological environments. [ABSTRACT FROM AUTHOR]

Copyright of Biomédica: Revista del Instituto Nacional de Salud is the property of Instituto Nacional de Salud of Colombia and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Our study aimed to develop an optimised laboratory protocol ensuring the preservation of morphological structures and extraction of high-quality DNA sequences from Psychodidae (Insecta, Diptera) specimens. With 310 analysed specimens, we investigated the impact of distinct laboratory treatments by employing two shaking categories (constant and interrupted) with five different incubation periods (16, 12, 8, 4 and 2 hours) during the DNA extraction process. Notably, 80.65% of the specimens exhibited morphological changes during DNA extraction. Our results indicated no statistical difference between constant and interrupted shaking for the total of morphological structures lost. However, within each shaking category, the loss of structures was influenced significantly by the incubation period. Prolonged incubation correlated with increased structural losses, whereas shorter incubation periods caused minor alterations in structures lost. In addition, our results showed a significant difference between constant and interrupted shaking treatments for DNA concentration. Likewise, the incubation period showed differences within each shaking category. Successful COI sequencing was achieved in 89.6% of specimens, with negligible differences in DNA fragment lengths across treatments. Our findings underscore the importance of an optimised protocol and its potential in systematic research involving nematoceran dipteran specimens by balancing morphological integrity and DNA extraction efficiency. [ABSTRACT FROM AUTHOR]

We report the first record of Edentomyia piauiensis in Maranhão State, Brazil in a cave, a similar ecotope where this species has been previously described in the literature. The specimens were captured in Toca do Inferno Cave, Barão de Grajaú, Maranhão, from September/2021, January, and May/2022. Twelve Hoover Pugedo (HP) light traps were installed at 1.5 m above the ground, uninterruptedly for 24h. We captured 1778 specimens of E. piauiensis. This finding expands knowledge of the geographical distribution of this species in Brazil. [ABSTRACT FROM AUTHOR]

A new species of Psychodidae is described by a well-preserved adult male trapped in Miocene amber from Simojovel, Chiapas, Mexico. The new species named Trichomyia vetusta sp. nov., is described, illustrated, and compared with the other Trichomyia fossil species from Mexican amber. [ABSTRACT FROM AUTHOR]

The main goal of the current study was to investigate the molecular systematics and host–vector interactions of sandfly species (Phlebotomus spp.) in Iran. Sandflies were collected using CDC light traps, sticky traps, and BG-Sentinel traps at seven trapping sites in Iran during 2019 and 2020. The source of blood meals was identified using primer pairs specific to either the 16S rRNA or the mitochondrial cytochrome b (cytb) genes. A total of 736 blood-fed female sandflies were collected. Six species of the genus Phlebotomus (Ph.) and four species of the genus Sergentomyia (Se.) were identified using molecular identification techniques. The most abundant sandfly species were Ph. papatasi (38.45 %) and Se. sintoni (36.28 %). Blood meal analysis identified 19 different host species, specifically, 15 mammalian and four bird species. The most common blood meal sources were Rattus norvegicus , followed by Homo sapiens , Bos primigenius f. taurus , Ovis orientalis f. aries , Meriones persicus , Gallus gallus f. domestica , Rhombomys opimus , and Canis lupus; other host species had incidences of less than 5 %. We observed that Ph. major complex and Se. sintoni had mostly mammalophilic blood-feeding behavior, while Ph. papatasi and Ph. sergenti had mostly anthropophilic and ornithophilic blood-feeding preferences, respectively. The current results confirmed cytb is a suitable gene for regular species level identification, and that phylogenetic analyses of cytb allowed for clear demarcation of sandfly species. [ABSTRACT FROM AUTHOR]

The present work assessed the experimental susceptibility of Nyssomyia antunesi and Lutzomyia longipalpis to Leishmania (Viannia) lainsoni and L. (V.) lindenbergi. A L. (Leishmania) chagasi–Lu. longipalpis combination was used as a susceptible control. Wild-caught Ny. antunesi and laboratory-bred Lu. longipalpis were membrane-fed on blood with a 5 × 106/mL log-phase promastigote culture suspension and dissected on days 2 and 8 post-blood meal (pbm) for analysis focused on the assessment of parasitoses, as well as placement and promastigote morphotyping. Survival curves were constructed. In all combinations, promastigotes were observed on day 8 pbm. For both Leishmania species, in Lu. longipalpis, the presence of parasites was observed up to the stomodeal valve, while in Ny. antunesi, the presence of parasites was observed up to the cardia. There were no significant differences in parasitosis between L. (V.) lainsoni and L. (V.) lindenbergi in either Ny. antunesi or Lu. longipalpis. Six morphological promastigote forms were distinguished in Giemsa-stained gut smears. The survival curves of all combinations decreased and were affected differently by several Lu. longipalpis–parasite combinations, as well with Lu. longipalpis–uninfected blood. These findings stress Lu. longipalpis as experimentally susceptible to Leishmania spp. and suggest the putative susceptibility of Ny. antunesi to L. (V.) lainsoni and L. (V.) lindenbergi. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Mosquitoes are of major public health importance throughout the world. Recently, there has been an increase in mosquito-borne diseases some of which are newly emerging, and some are remerging. Chemical control remains the main approach to reducing mosquito vector populations, therefore mitigating the transmission and spread of mosquito-borne diseases. Urban land uses have been identified as highly productive for container mosquitoes and thus require vector control interventions. In this study, we compared the effectiveness of the larvicide triflumuron in controlling mosquito vectors in households, cemeteries and tire shops and assessed its potential impact on non-target dipterans in urban temperate Argentina. On average, treatment with triflumuron reduced the occurrence of Aedes aegypti by 81% and Culex pipiens by 45%, with no significant differences among land uses. Two-thirds of the containers with these two mosquito species also harbored at least one non-target dipteran species (other Culicidae, Chironomidae, Psychodidae, Ceratopogonidae, Phoridae, Ephydridae, Syrphidae and Sciomyzidae). Triflumuron application also had unintended effects on non-target dipteran communities, resulting in reduced richness and diversity across all land uses, as well as a reduced density of individuals in tire shops. Triflumuron proved highly effective against Ae. aegypti and moderately effective against Cx. pipiens regardless of land-use patterns. However, unwanted effects against some non-target accompanying Diptera fauna were also recorded, suggesting a severe limitation for the use of this larvicide in areas where biodiversity conservation is of paramount importance. Preserving biodiversity and human welfare must be a primary goal in the search for better vector control measures. [ABSTRACT FROM AUTHOR]

Simple Summary: The old-world sand fly, Phlebotomus papatasi (Scopoli 1786), is a major vector of Leishmania major, the predominant pathogen responsible for zoonotic cutaneous leishmaniasis in the Middle East, North Africa, Southern Europe, and Central Asia. DEET and other synthetic insect repellents have been used for personal protection against sand fly bites. However, the frequent use of DEET repellent raised concerns in regards to skin sensitivity, toxicity, and unpleasant odor. There are increasing efforts to evaluate natural products for use in developing more effective organic sand fly repellents. This paper reports the results of a laboratory study on several plant essential oils and saturated fatty acids concerning their repellency against female sand flies. A static air repellency assay was used to measure the responses of sand flies to test materials. The sand fly repellency of each test material was compared with those achieved by commercial repellent DEET and IR35353 at the same test concentration. The study identified two of the tested essential oils as effective spatial repellents at reduced concentrations compared to those of DEET, and two saturated fatty acids were found to produce significant sand fly mortality. The results from this study establish the foundation for developing more effective natural sand fly repellent products. The sand fly, Phlebotomus papatasi (Scopoli, 1786), is a major vector for Leishmania major in the Middle East, which has impacted human health and US military operations in the area, demonstrating the need to develop effective sand fly control and repellent options. Here, we report the results of spatial repellency and avoidance experiments in a static air olfactometer using the female P. papatasi testing essential oils of Lippia graveolens (Mexican oregano), Pimenta dioica (allspice), Amyris balsamifera (amyris), Nepeta cataria (catnip), Mentha piperita (peppermint), and Melaleuca alternifolia (tea tree); the 9–12 carbon saturated fatty acids (nonanoic acid, decanoic acid, undecanoic acid, and dodecanoic acid); and the synthetic repellents DEET and IR3535. The materials applied at 1% exhibited varying activity levels but were not significantly different in mean repellency and avoidance from DEET and IR3535, except in regards to nonanoic acid. Some materials, particularly nonanoic and undecanoic acids, produced sand fly mortality. The observed trends in mean repellency over exposure time included the following: (1) P. dioica oil, M. alternifolia oil, decanoic acid, undecanoic acid, DEET, and IR3535 exhibited increasing mean repellency over time; (2) oils of N. cataria, A. balsamifera, M. piperita, and dodecanoic acid exhibited relatively constant mean repellency over time; and (3) L. graveolens oil and nonanoic acid exhibited a general decrease in mean repellent activity over time. These studies identified the essential oils of N. cataria and A. balsamifera as effective spatial repellents at reduced concentrations compared to those of DEET. Additional research is required to elucidate the modes of action and potential synergism of repellents and essential oil components for enhanced repellency activity. [ABSTRACT FROM AUTHOR]

Background:Phlebotomus papatasi (Diptera: Psychodidae) is the main vector of zoonotic cutaneous leishmaniasis. Wolbachia is a symbiotic alphaproteobacteria of arthropods that can be involved in susceptibility or resistance. This study aimed to investigate the relationship between Wolbachia and Deltamethrin susceptibility/resistance in Ph. papatasi. Deltamethrin filter papers (0.00002%) were used to test sand fly field collected from southern Iran. After the test, PCR amplification of the Wolbachia surface protein gene (wsp) was used to measure Wolbachia infection rate in the killed, surviving, and control groups. Result: The rates of infection by Wolbachia strain (wPap, super group A) differed between killed (susceptible) and surviving (resistant) Ph. papatasi specimens. The rate of Wolbachia infection in susceptible individuals was more than twice (2.3) (39% vs. 17%) in resistant individuals with the same genetic background. This difference was highly significant (p < 0.001), indicating a positive association between Wolbachia infection and susceptibility to Deltamethrin. In addition, the results showed that Deltamethrin can act as a PCR inhibitor during detection of Wolbachia in Ph. papatasi. Conclusion: Results of this study show that Wolbachia is associated with Deltamethrin susceptibility level in Ph. papatasi. Also, as Deltamethrin has been identified as a PCR inhibitor, great care must be taken in interpreting Wolbachia infection status in infected populations. The results of this study may provide information for a better understanding of the host-symbiont relationship, as well as application of host symbiosis in pest management. [ABSTRACT FROM AUTHOR]

Phlebotomine sandflies are an important group of insects from a medical perspective that are responsible for the transmission of leishmaniasis. The phenotypic and genotypic investigation of species of sand flies (Al-Harms) as the vector of cutaneous leishmaniasis was conducted in the Erbil province-Iraq for the first time between January and December 2022. Out of 2054 collected samples 1137 (55.4%) were male and 917 (44.6%) were female. statistically, there is no correlation between gender and sandfly species at (P<0.20). Three species were identified as belonging to the genus Phlepotomus: P. papatasi, P. sergenti, and P. Alexandria. P. papatasi was the most common species, with 1163 (15.6%), followed by P. sergenti (598 (29.1%), and P. alexandri 293(14.1%). There were two peaks of seasonal abundance of sand flies, the first peak of the seasonally abundant cutaneous leishmaniasis vectors occurred in May, while the second peak occurred in September, statistically, there is a correlation between the periods and sand fly species at (p<0.00). The city's health system must take the necessary precautions to control sand fly species as the vectors of cutaneous leishmaniasis disease. Controlling the sandflies will be a key to controlling (CL) disease. [ABSTRACT FROM AUTHOR]

Leishmaniases are important neglected tropical diseases caused by protozoans of the genus Leishmania spp. The female sandflies bite (Diptera: Psychodidae: Phlebotominae) is the main transmission form in the epidemiological chains of these diseases. Thus, studies on Phlebotominae subfamily are fundamental to improve the comprehension of the leishmaniases epidemiology, revealing relationships of these dipterans with the parasite and reservoirs. An entomological survey of Phlebotomine sandflies was carried out to investigate the fauna, Leishmania spp. DNA detection and blood meals sources of female sandflies captured in peridomiciles areas from a leishmaniases endemic area of São Paulo state, Brazil. A total of 637 Phlebotomine sandflies specimens were captured, and twelve species identified, some of them proven or suspected vectors of tegumentary leishmaniasis (TL) and visceral leishmaniasis (VL). Evandromyia (Aldamyia) carmelinoi and Lutzomyia (Lutzomyia) longipalpis PCR positive to Leishmania (Leishmania) infantum kinetoplast minicircle DNA (kDNA) were identified. Also, Leishmania (Leishmania) amazonensis kDNA was found in Evandromyia (Aldamyia) lenti and Lu. (Lu.) longipalpis. Regarding blood meals source, DNA of swine, humans, dogs, cattle, chickens, and opossums were detected in the midgut of Phlebotomine sandflies females captured in the study area. Our results highlight ecological relationships among different species of Phlebotomine sandflies, domestic and wild-synanthropic vertebrates (including humans) and two Leishmania species in peridomiciles from a leishmaniases endemic area of Brazil. [ABSTRACT FROM AUTHOR]

Turkey is among the countries where sand flies (Diptera: Psychodidae) are responsible for the endemic presence of leishmaniasis. Understanding the feeding behaviour of haematophagous arthropods is crucial for revealing the transmission cycles of vector-borne diseases. However, research on the host preferences of sand fly species in Turkey has been limited. This study aims to determine the blood meal sources of sand fly species collected from a wide range of geographical areas and to identify their feeding patterns. For the blood meal analysis, Sanger sequencing targeting the vertebrate mitochondrial cytochrome b or mammalian 12 S rRNA genes was performed on 265 female specimens collected between 2006 and 2019. The feeding patterns of sand flies were determined using Null model analysis. Host assignments of 218 individuals revealed nine different vertebrate hosts for 14 sand fly taxa. Bos taurus was the predominant host (81.19%), followed by Gallus gallus (8.88%), Capra hircus (5.14%), Mus musculus (1.81%), Ovis aries (0.93%), Homo sapiens (0.47%), Canis lupus familiaris (0.47%), Felis catus (0.47%) and Sitta spp. (0.47%). We discovered an aggregated feeding pattern among sand fly taxa instead of random or segregated feeding. Most sand fly taxa predominantly fed on bovines as their hosts, highlighting the need for further research on the role of domestic animals in the transmission of sand fly-borne diseases. Paraphlebotomus specimens displayed a relatively broad host range, particularly favouring Avian hosts, which requires further investigation into their life history traits. [ABSTRACT FROM AUTHOR]

Sand flies (Diptera: Psychodidae) are significant medical vectors of pathogens that cause illness in humans and animals. The most significant of these is the genus Leishmania (Kinetoplastida: Trypanosomatidae). Leishmaniasis is an important vector-borne disease. They are spread throughout the Old and New Worlds by the bites of female phlebotomine sand flies. Despite this importance, certain provinces where autochthonous leishmaniasis has historically existed and is re-emerging are lacking current data regarding their sand fly fauna. To provide a current update on the local sand fly fauna, a comprehensive entomological survey encompassing eight provinces was conducted subsequent to an analysis of past data regarding the species composition and distribution of sand flies in the area. Thus, the purpose of this study was to ascertain the species composition in various locations across eight provinces spread across four regions in Thailand. In order to gather information about the distribution of the phlebotomine sand fly, researchers used modified Centers for Disease Control and Prevention light traps between February 2021 and February 2022. At these 30 trappings in each province, we conducted 240 trapping nights in total. Collected sand flies were morphologically identified. The adult sand flies collected in this study were 8314 and represented 24 species of four genera, including 16 of Sergentomyia, 6 of Phlebotomus, 1 of Idiophlebotomus, and 1 of Chinius. Among these species, Sergentomyia anodontis had the largest population (25.71%), and population peaks were observed in Uthaithani province (51.81%). The collected sand flies' male-to-female ratio was 1.21:1.00 (4,556:3,758). The study also discovered that Phlebotomus argentipes, a vector of Leishmania spp., was captured in Phayao province, and Phlebotomus major major was majorly found in Uthaithani province. The widespread distribution of sand fly populations in this study may indicate the epidemiologic importance of vector ecology in this habitat. Our results confirm the presence of several sand fly species in different provinces of Thailand and provide valuable information about the distribution and behavior of phlebotomine sand flies, which can be used to develop effective control strategies for the prevention of sand fly-borne diseases. In several Thai provinces, the study also tracked the nocturnal movements of both male and female sand flies. The results showed that phlebotomine sand flies are more active at night, peaking just after dusk. [ABSTRACT FROM AUTHOR]

Background: Phlebotomus argentipes complex is the primary vector for cutaneous leishmaniasis, a burgeoning health concern in contemporary Sri Lanka, where effective vector control is important for proper disease management. Understanding the genetic diversity of the P. argentipes population in Sri Lanka is vital before implementing a successful vector control program. Various studies have indicated that genetic divergence, caused by genetic drift or selection, can significantly influence the vector capacity of arthropod species. To devise innovative control strategies for P. argentipes, exploring genetic diversity and phylogeography can offer valuable insights into vector competence, key genetic trait transfer, and impact on disease epidemiology. The primary objective is to analyze the genetic diversity and phylogeography of the P. argentipes complex in Sri Lanka, based on two mitochondrial genomic regions in modern representatives of P. argentipes populations. Methodology: A total of 159 P. argentipes specimens were collected from five endemic areas of cutaneous leishmaniasis and identified morphologically. Two mitochondrial regions (Cytochrome c oxidase subunit I (COI) and NADH dehydrogenase subunit 4 (ND4) were amplified using the total DNA and subsequently sequenced. Partial sequences of those mitochondrial genes were utilized to analyze genetic diversity indices and to explore phylogenetic and phylogeographic relationships. Principal findings: Among five sampling locations, the highest genetic diversity for COI and ND4 was observed in Hambantota (Hd—0.749, π—0.00417) and Medirigiriya (Hd—0.977, π—0.01055), respectively. Phylogeographic analyses conducted using COI sequences and GenBank retrieved sequences demonstrated a significant divergence of P. argentipes haplotypes found in Sri Lanka. Results revealed that they have evolved from the Indian ancestral haplotype due to historical- geographical connections of the Indian subcontinent with Sri Lanka. Conclusions: Utilizing high-mutation-rate mitochondrial genes, such as ND4, can enhance the accuracy of genetic variability analysis in P. argentipes populations in Sri Lanka. The phylogeographical analysis of COI gene markers in this study provides insights into the historical geographical relationship between India and P. argentipes in Sri Lanka. Both COI and ND4 genes exhibited consistent genetic homogeneity in P. argentipes in Sri Lanka, suggesting minimal impact on gene flow. This homogeneity also implies the potential for horizontal gene transfer across populations, facilitating the transmission of genes associated with traits like insecticide resistance. This dynamic undermines disease control efforts reliant on vector control strategies. [ABSTRACT FROM AUTHOR]

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. An integrated approach using phylogeographic data, climate modelling, and network analysis unlocks insights into the post-glacial expansion of the sand fly Phlebotomus mascittii in Europe and suggests two potential (parallel) dispersal routes. [ABSTRACT FROM AUTHOR]

Sandflies (Diptera; Psychodidae) are medical and veterinary vectors that transmit diverse parasitic, viral, and bacterial pathogens. Their identification has always been challenging, particularly at the specific and sub-specific levels, because it relies on examining minute and mostly internal structures. Here, to circumvent such limitations, we have evaluated the accuracy and reliability of Wing Interferential Patterns (WIPs) generated on the surface of sandfly wings in conjunction with deep learning (DL) procedures to assign specimens at various taxonomic levels. Our dataset proves that the method can accurately identify sandflies over other dipteran insects at the family, genus, subgenus, and species level with an accuracy higher than 77.0%, regardless of the taxonomic level challenged. This approach does not require inspection of internal organs to address identification, does not rely on identification keys, and can be implemented under field or near-field conditions, showing promise for sandfly pro-active and passive entomological surveys in an era of scarcity in medical entomologists. [ABSTRACT FROM AUTHOR]

Sandflies (Diptera: Psychodidae) are medically important insects prevalent in tropical and temperate regions of the world. About 30 species of these flies have been recorded from the Western Ghats region of Kerala. While carrying out epidemiological investigations on Leishmaniasis in this region, we found two species of field-collected sandflies viz., Phlebotomus argentipes and Sergentomyia monticola infected with Tylenchid nematode parasites. The infection rates were 0.89% and 0.9%, respectively. The parasite density of nematode juvenile stages was more than 1000 in all the specimens. Both males and females were found infected. We performed molecular characterization of these nematode parasites using multiple genetic markers, mitochondrial COI, and two different regions of 18S rDNA. The genetic analysis revealed that the nematode belonged to the genus Howardula of family Allantonematidae, one infesting P. argentipes (major vector species of Leishmania) and the other, S. monticola. This is the first report of natural infection with the entomo-parasitic nematode species, belonging to Allantonematidae among these sandfly species from India. Genetically related and unclassified species of nematodes belonging to this family had been reported elsewhere from termite species. As larval stages of sandflies develop in the organic matter of termite mounds, this finding may have significant implications on their bionomics and control. [ABSTRACT FROM AUTHOR]

Phylogenetic analyses between fossil species of the subfamily Bruchomyiinae were conducted. Based on them, a new subgenus, Palaeoglaesum (Amplissimum) subgen. nov., within the genus Palaeoglaesum is proposed. Two new species, Palaeoglaesum angustum sp. nov. and Palaeoglaesum cracoviae sp. nov., from Cretaceous Myanmar amber are described and illustrated herein. [ABSTRACT FROM AUTHOR]

Background: Phlebotomus papatasi (Diptera: Psychodidae) is the main vector of zoonotic cutaneous leishmaniasis (ZCL) in Iran. The nonstandard use of pesticides against pests, particularly in agriculture, indirectly has caused the development of resistance and, consequently, the threat of control measures in ZCL endemic areas. Up to 2023, several reports of resistance in Ph. papatasi have been declared in the Old World. The purpose of this study was to measure the lethal time (LT50 and LT90) of Ph. papatasi sand flies in the ZCL endemic center of Esfahan to DDT and deltamethrin insecticides. Methods: Sand flies were collected in Borkhar and were tested using WHO adult mosquito test kit against DDT 4% and deltamethrin 0.0002%. The sand fly’s survival was recorded during exposure time in 225, 450|, 900, 1800, and 3600- seconds’ intervals for DDT and Deltamethrin and they were allowed to recover for 24 hours. Then LT50 and LT90 were analyzed using probit software. Phlebotomus papatasi were identified using morphological keys and other sand flies’ species were excluded from the analysis. Results: The insecticide against female Ph. papatasi revealed hundred percent mortality when exposed to DDT 4% and deltamethrin 0.0002%. The LT50 and LT90 were 19.32 and 22.74 minutes for DDT 4% and 39.92 and 51.33 minutes for deltamethrin 0.0002% respectively. Conclusion: Results of this study revealed that Ph. papatasi is still susceptible to DDT and deltamethrin. This data provides valuable knowledge to implement effective control strategies against ZCL main vector and help to manage insecticide resistance in the region. [ABSTRACT FROM AUTHOR]

Abstract The geographical distribution of sand flies in Brazil has been the subject of some studies, yet there is no information about the phlebotomine fauna in João Pessoa, State of Paraíba, Brazil. The aim of this work is to evaluate the occurrence and distribution of sand flies in the Atlantic forest fragments and to evaluate a possible dispersion in 06 nearby districts. Light traps were used during three consecutive nights, supplemented by an aspirator during the dry period and rainy season. A total of 222 sand flies were found, 143 (130 males and 13 females) in the Atlantic forest, and 79 in urban areas. During the entire dry season, three species of phlebotomine sand flies were recorded in 11 forest fragments, Lutzomyia longipalpis, Lu. migonei and Lu. whitmani. During the rainy season, only Lu. longipalpis was found. This was the only species identified in the studied neighborhoods during both seasons. The differences in diversity of sand flies encountered between natural habitats and urban areas may thus be correlated mostly with adaptations to particular habitats and availability of food. One species (Lu. longipalpis) appears to be rapidly adapting to urban areas because of deforestation.

The recent geographic spread of Leishmania infantum along the borders of Argentina, Brazil and Paraguay has been highlighted. In our previous study, Lutzomyia longipalpis was found in 55 of 123 patches surveyed, and in some patches, sandflies were found at higher densities, forming hotspots. Based on the One Health approach, we investigated the seasonality of the vector, the presence of parasite DNA, and the environmental factors that contribute to vector and parasite dispersal in these previously described hotspots in Foz do Iguaçu, Brazil. Entomological surveys were conducted monthly for one year. Fourteen hotspots peridomicile and six intradomicile were sampled. PCR was used to assess the prevalence of Leishmania DNA in sandflies. Zero-inflated negative binomial regression was used to determine the association of micro- and mesoscale environmental variables with the occurrence and abundance of the three most abundant sandfly species sampled. A total of 3543 species were captured, with Lutzomyia longipalpis being the predominant species (71.78%) of the 13 species found. Evandromyia edwardsi, Expapillata firmatoi, Micropygomyia ferreirana and Pintomyia christenseni were reported for the first time in the region. NDVI, distance to water, precipitation, west-to-east wind, wind speed, maximum and minimum relative humidity, and sex were significant variables associated with vector presence/abundance in the environment. Vector presence/abundance in the peridomicile was associated with precipitation, altitude, maximum temperature, minimum and maximum relative humidity, west-to-east wind, wind speed, and sex. Leishmania DNA was detected in an average of 21% of Lu. longipalpis throughout the year. Vector abundance is concentrated in urban and peri-urban areas, with some specimens present in different parts of the city and some sites with high vector abundance. This distribution suggests that the risk of actual contact between humans and parasite vectors in urban areas during the epidemic period is associated with patches of peri-urban vegetation and then extends into urban areas.

Background: The successful use of semiochemicals to attract insects to traps is based on research on the most suitable compounds and their release profiles over time. Based on the group's promising results, matrices with a more adequate release profile and more eco-friendly properties for the release of 1-hexanol were developed. To use a more suitable prototype in the field, the most promising systems were added to a capsule and evaluated in a wind tunnel. Behavioral experiments were performed using the sand fly species, Lutzomyia longipalpis, to evaluate the efficacy of the proposed system. Methods: Different delivery systems were developed by varying the polymer (gellan gum and pectin) ratio, crosslinker (aluminum chloride) concentration, and glutaraldehyde removal.The delivery systems were loaded with 1-hexanol, and their release profiles were evaluated using gravimetric analysis under ambient and high-humidity conditions. When the matrix system was placed inside a plastic container, modulations in the active release profile were observed and the system could be reused. Actid attraction behaviors of the sand fly species, Lu. longipalpis, were evaluated in a wind tunnel when exposed to 1-hexanol-loaded release systems at different times. Results: Among the four formulations evaluated, System 2 (gellan gum and pectin in a 1:1 ratio with 5% aluminum chloride) exhibited the most promising release profile, with greater uniformity and longer compound release time. The maximum 1-hexanol release uniformity was achieved over a longer time, mainly every 24 h, under both ambient and high-humidity conditions. System 2 can be reused at least once with the same structure. The wind tunnel trials exhibited efficient activation and attraction of Lu. longipalpis to 1-hexanol after 24, 48, and 72 h in System 2 placed inside the capsules. Conclusions: The polymeric matrix supplemented with 1-hexanol and introduced in plastic capsules showed promising results in attracting sand flies. This system can be used as a solution for other attractive compounds as well as in other applications where their release needs to be controlled or prolonged. [ABSTRACT FROM AUTHOR]

Leishmaniasis is a serious public and veterinary health concern. Species of the subgenus Larroussius are the suspected and proven vectors of the leishmaniasis visceral form. This study was aimed at investigating the spatiotemporal dynamics of Phlebotomus perniciosus (Diptera: Psychodidae) and characterizing its habitats using satellite images. The entomological sampling was based on a preliminary stratification of the study area, in which the sandflies were collected at 12 selected stations in Fez City using sticky paper traps. The environmental factors studied for each station were land surface temperature (LST), normalized difference vegetation index (NDVI), and soil moisture index (SMI). Correlations between the number of species of sandflies and each of the environmental factors were evaluated using Pearson's correlation. Differences were considered significant at p < 0.05 (95% confidence level). Two species of the subgenus Larroussius, which are vectors of visceral leishmaniasis, Ph. perniciosus (96.54%) and Ph. longicuspis (3.46%), were found in the collected 231 specimens. Statistical analysis of the environmental factors also demonstrated a significant positive correlation between the number of Ph. perniciosus and LST in all environments. However, the correlation between the isolated species and NDVI was not strong and only significant in the urban area (r = 0.79, p = 0.0018), and no significant correlation was found with the SMI. The findings of this study could help health authorities develop appropriate future vector control strategies. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Background: 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. Main results: 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. Conclusions: 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. Author summary: Phlebotomine sand flies are tiny blood sucking insects. These are vectors of Leishmania, flagellate unicellular parasites, mostly found in Mediterranean and tropical areas. In Thailand, Leishmania martiniquensis and L. orientalis are responsible for local diseases but the parasite transmission cycle remains unknown. For a long time, South-Eastern Asia was considered to be free of leishmaniasis and as such, Phlebotomine sand flies have been understudied in this area. We carried out an inventory in a cave located in the Chiang Rai district, northern Thailand. We caught two new species we consider to be new for Science based on morphological (head and genital apparatus) and molecular characters (cytochrome b mitochondrial DNA sequences). The new species were named Phlebotomus shadenae n. sp. and Sergentomyia maiae n. sp. We also recorded the first species in Laos whereas the second has just been recorded in the studied cave. [ABSTRACT FROM AUTHOR]

Background: Zoonotic cutaneous leishmaniasis is a major public health problem in Iran with the main vector of Phlebotomus papatasi. The use of entomopathogenic fungi for biological control of the vector is a potential substitute for the current methods which are being used. The purpose of the current study was to assess the virulence of two local isolates of Beauveria bassiana (OZ2 and TV) against Ph. papatasi. Methods: To perform the bioassay test, fungal suspensions were applied for every stage of the sand fly life cycle. The mortality rate, longevity, and number of eggs laid were determined. Also, the probability of fungal survival on the surface of rodent’s body was assessed. Results: The longevity of infected adult sand flies with both isolates of B. bassiana was significantly lower (P< 0.05) in comparison to the negative control. The estimated Lethal concentration 50 (LC50) values for adult female and male sand flies treated with OZ2 isolate were 1.4×106 and 2.2×107 conidia/ml, respectively, while they were 6.8×106 and 2.3×108 conidia/ml for TV isolate, respectively. Both isolates of B. bassiana exhibited nonsignificant mortality rates in sand fly larvae and pupae and fecundity rate (P> 0.05). According to our findings for both isolates, the fungus continued to spread throughout the surface of the rodent's body for 144 hours after spraying. Conclusion: The current study demonstrated that both isolates of B. bassiana have considerable biological control capacity against adult sand flies. [ABSTRACT FROM AUTHOR]

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Background: Prior to this study, the moth flies in The Netherlands were represented by 61 species. Our findings derive from a citizen-science expedition in the Vondelpark in Amsterdam, one of the oldest public parks and best known parks in The Netherlands. The combination of citizen science and the exploration of a well-known urban park has allowed us to contribute to the knowledge of moth fly species present in The Netherlands. The findings from this study provide valuable insights into the distribution, taxonomy and genetic resources of Psychoda and Panimerus species, enhancing our understanding of insect biodiversity and promoting future research in this field. New information: Our study provides two new geographical records of the moth flies in The Netherlands, namely, Psychoda uniformata Haseman, 1907 and Panimerus maynei (Tonnoir, 1920) elevating the total number of species to 63. Furthermore, we provide re-descriptions of the females of Panimerus notabilis (Eaton, 1893) and P. goetghebueri (Tonnoir, 1919). Additionally, we make available for the first time, the sequence of the 5'-end of the cytochrome c oxidase subunit I (COI) gene or COI Barcodes for Panimerus notabilis, P. goetghebueri and P. maynei. These COI Barcodes serve as valuable tools for future species identification within the genus. [ABSTRACT FROM AUTHOR]

Some sandflies are vectors for Leishmaniasis, therefore knowing the fauna of the region is important in combating the disease. Sandflies were captured in Concórdia, SC, Brazil, and Leishmania spp. in captured females. Thirty-eight specimens of six different species were collected, one of which (Brumptomyia carvalheiroi) is reported for the first time in the south of the country. However, there were no samples of sandflies positive for Leishmania spp. Despite reports of allochthonous cases in the region studied, the agent is not yet circulating in vectors, but these represent a potential risk when new cases occur. [ABSTRACT FROM AUTHOR]

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. Methods: The 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. Results: The 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. Conclusions: The 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. [ABSTRACT FROM AUTHOR]

The Neotropical genus Alepia Enderlein, 1937 (Diptera, Psychodidae) is newly recorded in Australia. We present new geographical records for Alepia viatrix Jaume-Schinkel, Kvifte, Weele & Mengual, 2022, extending the range of this species by 18,000 km. We attribute these new Australian records to the likely unintentional introduction of A. viatrix through international bromeliad trade. This moth fly was found by school children working with insect taxonomists through an Australian citizen-science project, Insect Investigators. We describe and present for the first time high-resolution SEM pictures of the eggs of the genus Alepia. [ABSTRACT FROM AUTHOR]

Trichomyia is a cosmopolitan genus, and it is the only genus of Trichomyiinae to include extant species. In recent years, an increasing number of species belonging to the genus have been described, particularly from the Neotropics. However, phylogenetic studies of these species are lacking. In addition, few studies have focused on this group, and none has focused on it exclusively, making it difficult to classify objectively the high diversity known today. Herein, we present the first phylogenetic analysis of Trichomyia. We studied specimens from all continents to propose a phylogeny and used it as the basis of a new classification. Two clades proposed as genera were recovered. The canonical informal classification of Trichomyia into Groups A and B is discussed, and Gondwanotrichomyia is assigned a new status. Although we do not propose a subgeneric classification for Trichomyiinae, we provide some comments on the previously proposed subgenera based on our phylogeny. In addition, we report on the expanded distribution of Trichomyia festiva , Trichomyia gabia and Trichomyia imarui. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Sand fly species are traditionally identified using morphological traits, though this method is hampered by the presence of cryptic species. DNA barcoding is a widely used tool in the case of insects of medical importance, where it is necessary to know quickly which species are present in a transmission area. Here, we assess the usefulness of mitochondrial cytochrome c oxidase subunit I (COI) DNA barcoding as a practical tool for species identification, correct assignment of isomorphic females, and to evaluate the detection of cryptic diversity that occurs in the same species. A fragment of the COI gene was used to generate 156 new barcode sequences for sand flies from different countries of the Neotropical region, mainly Colombia, which had been identified morphologically as 43 species. The sequencing of the COI gene allowed the detection of cryptic diversity within species and correctly associated isomorphic females with males identified by morphology. The maximum intraspecific genetic distances ranged from 0 to 8.32% and 0 to 8.92% using uncorrected p distances and the Kimura 2-parameter (K2P) model, respectively. The minimum interspecific distance (nearest neighbor) for each species ranged from 1.5 to 14.14% and 1.51 to 15.7% using p and K2P distances, respectively. Three species had more than 3% maximum intraspecific distance: Psychodopygus panamensis, Micropygomyia cayennensis cayennensis, and Pintomyia evansi. They also were split into at least two molecular operational taxonomic units (MOTUs) each, using different species delimitation algorithms. Regarding interspecific genetic distances, the species of the genera Nyssomyia and Trichophoromyia generated values lower than 3% (except Nyssomyia ylephiletor and Ny. trapidoi). However, the maximum intraspecific distances did not exceed these values, indicating the presence of a barcode gap despite their proximity. Also, nine sand fly species were DNA barcoded for the first time: Evandromyia georgii, Lutzomyia sherlocki, Ny. ylephiletor, Ny. yuilli pajoti, Psathyromyia punctigeniculata, Sciopemyia preclara, Trichopygomyia triramula, Trichophoromyia howardi, and Th. velezbernali. The COI DNA barcode analysis enabled the correct delimitation of several Neotropical sand fly species from South and Central America and raised questions about the presence of cryptic species for some taxa, which should be further assessed. [ABSTRACT FROM AUTHOR]

Phlebotomus argentipes and Phlebotomus colabaensis are the two sandfly species prevalent in the Western Ghats region of Kerala. P. argentipes had been incriminated as the vector of leishmaniasis in this region. The similar taxonomic features shared by these two species of sandflies had made it difficult to distinguish the adults morphologically. DNA barcoding is an efficient molecular diagnostic method utilized to identify the closely related species. In the present study, we designed species specific DNA primers from the DNA barcode sequences of P. argentipes and P. colabaensis. A semi nested PCR was performed with a consensus forward primer and two different reverse primers which could amplify specific DNA fragments for these two different sandflies. An amplified PCR product of 465 bp indicated P. argentipes and 401 bp, P. colabaensis. This species-specific PCR assay can be utilized to identify and distinguish two common Phlebotomus species with a single-step multiplex PCR. [ABSTRACT FROM AUTHOR]