Vector Of Leishmaniasis Research Articles

Impact of climate and land use on the temporal variability of sand fly density in Sri Lanka: A 2-year longitudinal study.

Leishmaniasis has emerged as an escalating public health problem in Sri Lanka, with reported cases increasing nearly three folds over past decade, from 1,367 in 2014 to 3714 cases in 2023. Phlebotominae sand flies are the vectors of leishmaniasis. Their density is known to be influenced by context-specific climatic and land use patterns. Thus, we aimed to investigate how these factors drive sand fly density across Sri Lanka. We analysed monthly collections of sand flies (n = 38,594) and weather data from ten sentinel sites representing three main geo-climatic zones across Sri Lanka, over 24 months. Site-specific land use data was also recorded. The influence of climate and land use patterns on sand fly density across the sentinel sites were estimated using distributed lag non-linear models and machine learning. We found that climate played a major role on sand fly density compared to land use structure. Increase in rainfall and relative humidity at real time, and ambient temperature and soil temperature with a 2-month lag were associated with a statistically significant increase in sand fly density. The maximum relative risk (RR) observed was 3.76 (95% CI: 1.58-8.96) for rainfall at 120 mm/month, 2.14 (95% CI: 1.04-4.38) for relative humidity at 82% (both at real time). The maximum RR was 2.81 (95% CI: 1.09-7.35) for ambient temperature at 34.5°C, and 11.6 (95% CI, 4.38-30.76) for soil temperature (both at a 2-month lag). The real-time increase in ambient temperature, sunshine hours, and evaporation rate, however, reduced sand fly density homogeneously in all study settings. The high density of chena and coconut plantations, together with low density of dense forests, homesteads, and low human footprint values, positively influenced sand fly density. The findings improve our understanding of the dynamic influence of environment on sand fly densities and spread of leishmaniasis. This knowledge lays a foundation for forecasting of sand fly densities and designing targeted interventions for mitigating the growing burden of leishmaniasis among the most vulnerable populations, particularly in an era of changing climate.

Primeiro registro de Lutzomyia longipalpis (Lutz e Neiva, 1912) em Macapá, Amapá

Lutzomyia longipalpis (Lutz & Neiva, 1912) constitutes the most epidemiologically relevant vector of visceral leishmaniasis (VL) in the New World. On October 25, 2023, the Macapá Center for Strategic Information in Health Surveillance registered a case of VL in the Km9 neighborhood, in Macapá. This study aimed to describe the Phlebotominae species in this area to assist the confirmation of the autochthony of the case. In total, 12 specimens were collected, of which five belonged to the Lutzomyia longipalpis species, confirming the presence of the VL agent vector and the possible autochthony of the transmission.

Chaetotaxy of the fourth larval stage of Pintomyia longiflocosa, a primary vector of cutaneous leishmaniasis in Colombia.

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. 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. 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. 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. 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.

A systematic review and meta-analysis of the prevalence of Leishmania infantum in sandflies in Iran

Objective: To determine the overall and pooled prevalence of Leishmania (L.) infantum in sandfly vectors in Iran. Methods: The present research conducted a systematic review and meta-analysis and searched regional databases such as PubMed, Scopus, Web of Science (WoS), Embase, PAHO Iris, LILACS, WHO Iris, and local databases named: SID, Magiran, Civilica, and also grey literatures. The current research included studies that were conducted in Iran and examined L. infantum in different sandfly vectors. The studies’ quality assessment/risk of bias assessment was evaluated by the Joanna Briggs Institute Critical Appraisal Checklist for prevalence data studies, and the data were analyzed by Stata 14 software. In addition, we examined 22 primary studies to estimate the overall prevalence of L. infantum among various vectors of visceral leishmaniasis. Results: According to the meta-analysis, the pooled prevalence of Phlebotomus (Ph.) tobbi, Ph. alexandri, Ph. kandelaki, Ph. perfiliewi, Ph. major, Ph. keshishiani were 5.34%, 4.36%, 2.23%, 1.79%, 4.37% and 1.18%. Ph. tobbi has the highest infection rate (25.00%) of L. infantum among the sandfly vectors. Conclusions: Visceral leishmaniasis is widespread in Fars, Ardebil, and East-Azerbaijan provinces, which are the most important endemic regions in Iran.

DIVERSITE SPECIFIQUE DES PHLEBOTOMES DU NIGER ET DE SES PAYS LIMITROPHES

Introduction: Cutaneous leishmaniasis is an endemic parasitosis in Niger and some of its neighbouring countries. The vectors of this disease are sandflies, which are unfortunately little known in Niger and some of its neighbouring countries. The aim of this study is to inventory the species of sandfly present in Niger and its neighbouring countries. Methodology: This study is conducted in two parts. The first is a systematic review and meta-analysis of articles published on sandflies in Niger and its neighboring countries between the period 2012 to 2022 through Ajol, Pubmed and Science direct. The second is based on a prospective collection of sandflies carried out in the town of Zinder, using castor oil and CDC light traps and morphologically identified. Results: All the sandflies recorded in the systematic review belong to the genera Phlebotomus and Sergentomyia, with a species richness of 43. Phlebotomusperniciosus predominates (28%) on the Maghreb pole. However, on the sub-Saharan pole, Sergentomyiaclydei predominated with 53%. A strong association between the Maghreb pole and the Phlebotomus genus was observed (p-value<0.05). The prospective study carried out in Zinder provided six species with Phlebotomuspapatasi (6%), Phlebotomusduboscqi (6%), Phlebotomussergenti (25%), Sergentomyiaschwetzi (6%), Sergentomyiaafricana (38%) and Sergentomyiaantennata (19%). Conclusion: Niger and its surroundings contain a wide variety of sandflies, with a dominance of species of the Phlebotomus genus, which are the main vectors of leishmaniasis. This study also reports the presence of six species in Zinder, including P. papatasi and S. schwetzi, newly captured in Niger.

Salivary antigens rPagSP02 and rPagSP06 are a reliable composite biomarker for evaluating exposure to Phlebotomus argentipes in Sri Lanka

Phlebotomus argentipes is the established vector of leishmaniasis in the Indian sub-continent. Antibodies to sand fly salivary antigens are biomarkers for vector-host exposure in leishmaniasis-endemic regions. Ph. argentipes transmits Leishmania donovani in Sri Lanka, primarily causing cutaneous leishmaniasis (CL). Our study compared the performance of salivary gland homogenate (SGH) from a lab-reared local strain of Ph. argentipes females to a composite recombinant salivary biomarker (rPagSP02 + rPagSP06) in a CL-endemic population. Sera from 546 healthy individuals, 30 CL patients, and 15 non-endemic individuals were collected. Western blot analysis of Ph. argentipes SGH identified immunogenic bands between 15 kDa and 67 kDa, with bands of predicted molecular weight ∼of 15 kDa (SP02) and ∼28–30 kDa (SP06) as the major antibody targets. Indirect ELISAs using SGH or rPagSP02 + rPagSP06 antigens showed high sensitivity (96.7%) and specificity (100%), detecting comparable seropositivity in endemic populations. rPagSP02 + rPagSP06 exhibited enhanced discriminatory ability, supported by a strong positive correlation (r = 0.869) with SGH. Our findings indicate that the composite rPagSP02 + rPagSP06 salivary biomarker effectively identifies Ph. argentipes exposure in individuals living in Sri Lanka, showing promising potential for use in surveillance. These findings should be further validated to confirm the epidemiological applications in leishmaniasis-endemic regions.

Seasonal dynamics, Leishmania diversity, and nanopore-based metabarcoding of blood meal origins in Culicoides spp. in the newly emerging focus of leishmaniasis in Northern Thailand

BackgroundClinical cases of leishmaniasis caused by Leishmania (Mundinia) parasites have been increasingly reported in Southeast Asia, particularly Thailand. Recent evidence has shown that Leishmania (Mundinia) parasites successfully developed into infective metacyclic promastigotes in Culicoides biting midges, strongly supporting their putative role in disease transmission. However, Culicoides diversity, host preference, and Leishmania prevalence in endemic areas remain largely unknown.MethodsWe investigated the seasonal dynamics, infection prevalence, and blood meal identification of Culicoides collected from the emerging focus of visceral leishmaniasis in Lampang Province, Northern Thailand, during 2021–2023. Midge samples were molecularly screened for Leishmania using SSU rRNA-qPCR and ITS1-PCR, followed by Sanger plasmid sequencing, and parasite haplotype diversity was analyzed. Host blood meal origins were comparatively identified using host-specific Cytb-PCRs and a nanopore-based metabarcoding approach.ResultsA total of 501 parous and gravid females and 46 blood-engorged ones belonging to at least 17 species of five subgenera (Remmia, Trithecoides, Avaritia, Hoffmania, and Meijerehelea) and two species groups (Shortti and Calvipalpis) were collected with temporal differences in abundance. Leishmania was detected by SSU rRNA-qPCR in 31 samples of at least 11 midge species, consisting of Culicoides oxystoma, C. guttifer, C. orientalis, C. mahasarakhamense, C (Trithecoides) spp., C. innoxius, C. shortti, C. arakawae, C. sumatrae, C. actoni, and C. fulvus, with the overall infection prevalence of 5.7%. The latter six species represent the new records as putative leishmaniasis vectors in Northern Thailand. The ITS1-PCR and plasmid sequencing revealed that Leishmania martiniquensis was predominantly identified in all qPCR-positive species, whereas L. orientalis was identified only in three C. oxystoma samples. The most dominant haplotype of L. martiniquensis in Thailand was genetically intermixed with those from other geographical regions, confirming its globalization. Neutrality test statistics were also significantly negative on regional and country-wide scales, suggesting rapid population expansion or selective sweeps. Nanopore-based blood meal analysis revealed that most Culicoides species are mammalophilic, with peridomestic and wild mammals (cow, pig, deer, and goat-like species) and humans as hosts, while C. guttifer and C. mahasarakhamense fed preferentially on chickens.ConclusionsThis study revealed seasonal dynamics and sympatric circulation of L. martiniquensis and L. orientalis in different species of Culicoides. Evidence of human blood feeding was also demonstrated, implicating Culicoides as putative vectors of human leishmaniasis in endemic areas. Further research is therefore urgently needed to develop vector control strategies and assess the infection status of their reservoir hosts to effectively minimize disease transmission.Graphical

Association between the visceral leishmaniasis vector Lutzomyia longipalpis and tree families in a Brazilian tropical urban area

BackgroundVisceral leishmaniasis (VL) is a neglected disease endemic to many tropical and subtropical countries. In Brazil, VL is caused by the protozoan parasite Leishmania infantum and is transmitted by the sand fly Lutzomyia longipalpis. The geographical variation in the incidence of the disease in certain urban areas of Brazil suggests a possible association with features of the urban landscape. The objective of this study was to verify the association of urban tree families with the presence of the vector Lu. longipalpis.MethodsThe study area of the municipality of Teresina, capital of the State of Piauí. The sand flies were captured using CDC light traps—one placed close to the houses, on a tree in the backyard, less than 5 m from the house, and another in the living room. The traps remained three consecutive nights, or, at the most, alternating days, working 12 h a night (from 6 p.m. to 6 a.m.), during 2016 and 2017.ResultsTree species were identified in an area of 100 m radius around CDC-type traps for capturing Lu. longipalpis installed in household backyards. Association between tree families and the occurrence of Lu. longipalpis was evaluated using the Fisher exact test and multiple correspondence analysis. Both Anacardiaceae (represented by mango and cashew trees) and Meliaceae (neem) families were more commonly found around sand fly traps capturing fewer Lu. longipalpis. Conversely, Fabaceae (several species of legumes) and Myrtaceae (guava) trees were associated with traps capturing higher number of VL vectors. In multiple correspondence analysis, the presence of trees from the Myrtaceae, Combretaceae (Almond), Fabaceae and Arecaceae (different species of palm trees) families were grouped with traps capturing high number of insects.ConclusionsThe findings suggest that trees from the Fabaceae, Myrtaceae, Combretaceae, and Arecaceae families might potentially attract sand flies, and Anacardiaceae and Meliaceae families may have a repellent effect. Finer analyses to distinguish the specific effects from the different species of the Anacardiaceae (mango and cashew), Arecaceae and Fabaceae families are still needed. The manipulation of urban landscaping might be a useful and clean alternative to help in the control of urban VL.

Improved high quality sand fly assemblies enabled by ultra low input long read sequencing

Phlebotomine sand flies are the vectors of leishmaniasis, a neglected tropical disease. High-quality reference genomes are an important tool for understanding the biology and eco-evolutionary dynamics underpinning disease epidemiology. Previous leishmaniasis vector reference sequences were limited by sequencing technologies available at the time and inadequate for high-resolution genomic inquiry. Here, we present updated reference assemblies of two sand flies, Phlebotomus papatasi and Lutzomyia longipalpis. These chromosome-level assemblies were generated using an ultra-low input library protocol, PacBio HiFi long reads, and Hi-C technology. The new P. papatasi reference has a final assembly span of 351.6 Mb and contig and scaffold N50s of 926 kb and 111.8 Mb, respectively. The new Lu. longipalpis reference has a final assembly span of 147.8 Mb and contig and scaffold N50s of 1.09 Mb and 40.6 Mb, respectively. Benchmarking Universal Single-Copy Orthologue (BUSCO) assessments indicated 94.5% and 95.6% complete single copy insecta orthologs for P. papatasi and Lu. longipalpis. These improved assemblies will serve as an invaluable resource for future genomic work on phlebotomine sandflies.

Risk factors of visceral leishmaniasis in the world: a review

Visceral leishmaniasis is a zoonotic parasitic disease caused by viscerotropic Leishmania species and transmitted by bites of infected phlebotomine sandflies, which is predominantly prevalent in the Indian subcontinent, eastern Africa and South America. Currently, visceral leishmaniasis is the second most fatal parasitic disease in the world. Because of climate changes, urban development and individual conditions, there are changes in the density of visceral leishmaniasis vector sandflies and the likelihood of contact with humans, resulting in a visceral leishmaniasis transmission risk. The review summarizes natural, social and biological factors affecting the transmission of visceral leishmaniasis, so as to provide insights into formulation of targeted control measures for visceral leishmaniasis.

Distribution and seasonal fluctuation of visceral leishmaniasis vectors sandflie in Lüliang City of Shanxi Province in 2023

To investigate the distribution and seasonal fluctuations of visceral leishmaniasis vectors sandflies in Lüliang City, Shanxi Province, so as to provide insights into assessment of the visceral leishmaniasis transmission risk and formulation of visceral leishmaniasis control measures. A total of 12 natural villages were sampled from Shilou County, Lishi District, Lanxian County, Linxian County and Wenshui County in Lüliang City, Shanxi Province from June to September, 2023, and sandflies were captured using light traps from 7 breeding habitats, including farmers' houses, sheep pens, cattle pens, chicken coops, pig pens, mule and horse pens, and loess-cave dwellings. Following morphological identification of the sandfly species, the distribution of sandflies and the seasonal fluctuations of the sandfly density were analyzed. In addition, the Leishmania was detected in sandflies using a real-time fluorescence quantitative PCR assay. A total of 2 831 sandflies were captured with 156 light traps in Lüliang City from June to September, 2023, including 2 638 female sandflies (93.18%) and 193 male sandflies (6.82%), and the average density was 16.91 sandflies/(light-night). The seasonal fluctuations of the sandfly density all appeared a unimodal distribution in all survey sites, and the sandfly density peaked in July and then declined rapidly. Among all types of breeding habitats, the greatest sandfly density was found in sheep pens [39.04 sandflies/(light-night)]. In addition, 4.08% (2/49) of the sandfly samples were tested positive for Leishmania nucleic acid as revealed by the real-time fluorescence quantitative PCR assay. Sandflies were widely distributed in Lüliang City, Shanxi Province in 2023, and the peak of the sandfly density was observed in July, which had a visceral leishmaniasis transmission risk. Intensified surveillance of visceral leishmaniasis and sandfly vectors is required and targeted vector control is recommended.

Distribution and seasonal fluctuation of visceral leishmaniasis vectors sandflies in Henan Province in 2023

To investigate the geographical distribution and seasonal fluctuations of visceral leishmaniasis vectors sandflies in Henan Province in 2023, so as to provide insights into the prevention and control of visceral leishmaniasis vectors. A total of 23 counties (districts) were sampled from 18 cities of Henan Province from May to September, 2023 as sandfly surveillance sites, and sandflies were captured using human capture and light trapping methods. Following morphological identification, the changes in the sandfly density were calculated at different months and in different breeding habitats. A total of 406 light traps were set at sandfly surveillance sites in Henan Province from May to September, 2023, and a total of 3 137 female sandlies were captured, with an average density of 7.73 sandlies/(light·night). A total of 1 494 Phlebotomus chinensis sandflies were captured, including 1 222 female sandflies, with an average density of 3.01 sandflies/(light·night), and the highest density of P. chinensis was found in Gongyi City [17.00 sandflies/(light·night)]. A total of 5 544 sandflies were captured using the human capture method, including 230 P. chinensis, and the density of P. chinensis appeared a unimodal distribution, with a peak in early July [5.81 sandflies/(light·night)]. Among different breeding habitats, the highest P. chinensis density was detected in pigpens [4.50 sandflies/(light·night)]. P. chinensis was predominantly distributed in hilly areas of northern and central-western Henan Province in 2023, and the sandfly density appeared a unimodal distribution. Intensified monitoring of visceral leishmaniasis vectors is recommended.

RPagSP02+rPagSP06 recombinant salivary antigen is a reliable biomarker for evaluating exposure to Phlebotomus argentipes in Sri Lanka.

Phlebotomus argentipes is the established vector of leishmaniasis in the Indian sub-continent. Antibodies to sand fly salivary antigens are biomarkers for vector-host exposure in leishmaniasis-endemic regions. Ph. argentipes transmits Leishmania donovani in Sri Lanka, primarily causing cutaneous leishmaniasis (CL). Our study compared the performance of salivary gland homogenate (SGH) from a lab-reared local strain of Ph. argentipes females to a composite recombinant salivary biomarker (rPagSP02 + rPagSP06) in a CL-endemic population. Sera from 546 healthy individuals, 30 CL patients, and 15 non-endemic individuals were collected. Western blot analysis of Ph. argentipes SGH identified immunogenic bands between 15 kDa and 67 kDa, with bands of predicted molecular weight õf 15 kDa (SP02) and ~28-30 kDa (SP06) as the major antibody targets. Indirect ELISAs using SGH or rPagSP02 + rPagSP06 antigens showed high sensitivity (96.7%) and specificity (100%), detecting comparable seropositivity in endemic populations. rPagSP02 + rPagSP06 exhibited enhanced discriminatory ability, supported by a strong positive correlation (r = 0.869) with SGH. Our findings indicate that the composite rPagSP02 + rPagSP06 salivary biomarker effectively identifies Ph. argentipes exposure in individuals living in Sri Lanka, showing promising potential for use in surveillance. These findings should be further validated to confirm the epidemiological applications in leishmaniasis-endemic regions.

Comparative Analysis of Phlebotomus argentipes Vector of Leishmaniasis in India and Sri Lanka.

Phlebotomus argentipes is the predominant sandfly vector of leishmaniasis in the Indian subcontinent. India and Sri Lanka primarily report visceral and cutaneous leishmaniasis caused by Leishmania donovani. We compared Ph. argentipes from two locations, focusing on its morphological, molecular, and salivary protein characteristics. Sandflies were captured using CDC light traps and cattle-baited net traps. Species identification and morphological comparisons were carried out using standard taxonomic keys. DNA extracted from 12 Sri Lankan sandfly samples was PCR-amplified and sequenced for the variable region of Cytochrome oxidase subunit I. Existing DNA sequences of India from GenBank were utilized for a phylogenetic analysis between Sri Lanka and India. Salivary protein profiles were studied using SDS-PAGE, Western blot, and electrospray ionization/LC/MS/MS. The morphological similarities observed between female Ph. argentipes from India and Sri Lanka suggest the presence of Ph. argentipes var. glaucus. A phylogenetic analysis showed genetic divergence between Ph. argentipes populations, but both shared a similar salivary protein profile. A common, strong 30 kDa immunogenic band comprised PagSP05, PagSP06, and PagSP17 proteins of Ph. argentipes. The similarity between the immunogenic salivary proteins suggests their potential use as common markers for vector exposure or immune response stimulants across regions. The use of multiple samples for each category of serum would improve the comprehensiveness of the immunogenic profiles obtained.

Spatio-Temporal distribution of a vector of cutaneous leishmaniasis: Pintomyia longiflocosa, in a population from the Colombian Andean Mountains.

Leishmaniasis, a neglected disease and public health concern, is associated with various factors such as biological, social, economical conditions and climate, increasing the risk of human infection. Understanding the population dynamics of the vectors, like Pintomyia longiflocosa, and its relationship with ecological variables is crucial for developing effective strategies to control sand fly populations and combat cutaneous leishmaniasis in a tropical country like Colombia. Adult sand flies were collected in three different sample locations: outdoor, indoor, and peri-domestic areas in three houses located in the rural settlement of Campoalegre (Huila) between February 2020 and February 2021, using the CDC light traps. The sand fly density was quantified and associated with the sample locations and the sampling months using Analysis of Variance and Pearson correlations. In the period of the sample, 98.86% of sand fly collected was identified as Pi. longiflocosa. The density of this species was significantly different between males and females, the latter contributing more to density in all sample locations (P<0.0001). The outdoor was the sample location with the highest and most significative density in this study (70%, P = 0.04). The density of these sand flies is related to the seasonality of Campoalegre, revealing a density peak from February and June to October (P < 0.05). Finally, precipitation is the environmental variable prominently linked to the density pattern, showing a negative correlation with it. Months with the highest precipitations show the lowest values of Pi. longiflocosa abundance. Our investigation reveals a inverse correlation between precipitation levels and the abundance of Pi. longiflocosa in Campoalegre (Huila), particularly in outdoor areas. This suggests that vector control strategies to periods of reduced precipitation in outdoor settings could offer an effective approach to minimizing cases of cutaneous leishmaniasis in the region.

A Comprehensive Review of the Situation of Visceral Leishmaniasis Vectors in Iran.

This study's major aim is to investigate the situation of visceral leishmaniasis vectors, with a focus on their distribution and relationships to the disease in Iran and some other old-world nations. The terms Iran and the Old World, along with the keywords sand flies, vectors, visceral leishmaniasis, distribution, and Phlebotomus, were searched in electronic databases from 1930 to 2018, including Pub-Med, Web of Science, Google Scholar, and MEDLINE. According to the findings, Phlebotomus tobbi was a mountain species, but it was also found in the plains, rodent nests, and rock crevices. This species was considered to be one of the vectors of visceral leishmaniasis in Iran. Phlebotomus kandelakii has been caught in Afghanistan, Iran, Lebanon, Turkey and Georgia. In Iran, the first infection of Phlebotomus kandelakii and Ph. perfiliewi transcaucasicus with Leishmania infantum were reported in northwest of Iran. Phlebotomus major was one of the complex sand flies, and its members include Ph. major (India, Nepal, and Pakistan), Ph. neglectus (Southern Europe, Crimea and Iran), Ph. syriacus (Southwest Asia, Caucasus), Ph. notus (Afghanistan and Iran), Ph. wenyoni (Iran and Iraq), and Ph. wui (China). The first natural infection of Ph. alexandri with Leishmania infantum was reported in 2006 in endemic foci of visceral leishmaniasis in Fars Province, southern Iran. The findings of this project suggest that sand fly species of the subgenus Larrousisus play a key role in the transmission of all types of visceral leishmaniasis in Iran.

Distribution and ecological aspects of sand fly species from Kousseri, an endemic focus of visceral leishmaniasis in Northern Cameroon

IntroductionKousseri in the Far North Region of Cameroon has long been known as an endemic focus of visceral leishmaniasis (VL), although the study on sand flies in this focus is scarce. The present study investigates the spatial distribution, seasonality, and ecological aspects of sand flies from Kousseri. This study is based on the need to optimize the effectiveness of leishmaniasis control programs in the northern part of the country. MethodsSand flies were sampled monthly over 12 months in five selected sites using CDC light traps. Only captured females were morphologically identified at species level based on valid keys. Results and discussionOverall, 4,214 sand fly specimens were collected during 360 trapping nights. The male/female sex ratio slightly favored females (1:1.04). The eudominant Sergentomyia antennata followed by both dominant Sergentomyia schwetzi and Sergentomyia squamipleuris were the most common and abundant species, accounting for 76.1% of the collection. Phlebotomus duboscqi, the confirmed vector of cutaneous leishmaniasis (CL) in West African foci, although rare in the collection, was found in four of the five surveyed sites. This sand fly species with Se. schwetzi abounded in peri-urban areas and, respectively, in animal shed and in uninhabited house biotopes, while Se. antennata and Se. squamipleuris prevailed in rural areas and, respectively, in animal shelter biotopes and outside dwellings. All caught sand fly species except Se. schwetzi, Sergentomyia clydei, Sergentomyia inermis, and Sergentomyia adleri prevailed during the dry season. The highest Shannon–Wiener index of sand flies due to the maximal richness and evenness was found in the urban area, in outdoor biotopes, and during the rainy season (H′ = 1.68, 1.80, and 1.74, respectively). These data provide evidence that less urbanized areas, animal shelters around the compounds, and the absence of precipitation (dry season) favored the dispersion of abundant sand fly species in Kousseri. Based on previous reports on Leishmania transmission, a surveillance plan is required to prevent an outbreak of VL or an establishment of CL or canine leishmaniasis (CnL) in this focus. Further research identifying the blood meal source and the Leishmania parasites in these insects is critical for providing insightful data to fight leishmaniasis in Northern Cameroon.

Susceptibility of Phlebotomus papatasi (Diptera: Psychodidae) against DDT and Deltamethrin in an Endemic Focus of Zoonotic Cutaneous Leishmaniasis in Iran.

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. 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. 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. 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.

First Molecular Identification of Phlebotomine Sandflies (Diptera: Psychodidae) in Nineveh Governorate, Northern of Iraq

Abstract Background: The dipteran phlebotominae sandflies are considered a major threat to public health because they are proven vectors of leishmaniasis, phleboviruses, and some species of pathogenic bacteria. Therefore, scientists paid great attention to studying it at the phenotypic and molecular levels. In Iraq, leishmaniasis is considered an endemic disease and exists in its visceral and cutaneous forms (Baghdad boil). Its spread is greater in the southern regions than in the northern regions due to high temperatures, which in turn leads to an increase in the widespread of the sandflies. Given the lack of studies in the northern region of Iraq, this study was conducted and included a survey of all provinces of the Nineveh Governorate (the second governorate in Iraq in terms of population density). Methods: The sandflies collection process was performed during the period from May 2023 to February 2024, and DT150-TUN light traps were used for this purpose. For the initial phenotypic identification, several taxonomic keys of the subfamily phlebotomine were used as male genitalia, pharynx, and female spermatheca. Conventional polymerase chain reaction based on the amplification of the COI gene was performed for the molecular process of 60 captured sandflies from about 34 regions and 89 sites. Before performing a phylogenetic tree, high-quality sequences are matching with the closest global isolates in the National Center for Biotechnology Information (NCBI). Results: A total of 4861 (2034 males and 2827 females) sandflies were captured in the current study and five species were identified belong to two genera (Phlebotomus and Sergentomyia). Sandflies were found to be more widespread in Mosul province and Phlebotomus papatasi was the most widespread of the rest of the species. Molecular results confirmed the other results of the study, and isolates of the identified species were recorded in NCBI. Conclusion: The current study demonstrated the importance of sandflie identification using molecular methods and was able for the first time from molecular identification of three species in Iraq. It is also the first molecular study of sandflies at the level of the Nineveh Governorate

Rhabdoviral Endogenous Sequences Identified in the Leishmaniasis Vector Lutzomyia longipalpis Are Widespread in Sandflies from South America.

Sandflies are known vectors of leishmaniasis. In the Old World, sandflies are also vectors of viruses while little is known about the capacity of New World insects to transmit viruses to humans. Here, we relate the identification of RNA sequences with homology to rhabdovirus nucleocapsids (NcPs) genes, initially in the Lutzomyia longipalpis LL5 cell lineage, named NcP1.1 and NcP2. The Rhabdoviridae family never retrotranscribes its RNA genome to DNA. The sequences here described were identified in cDNA and DNA from LL-5 cells and in adult insects indicating that they are transcribed endogenous viral elements (EVEs). The presence of NcP1.1 and NcP2 in the L. longipalpis genome was confirmed in silico. In addition to showing the genomic location of NcP1.1 and NcP2, we identified another rhabdoviral insertion named NcP1.2. Analysis of small RNA molecules derived from these sequences showed that NcP1.1 and NcP1.2 present a profile consistent with elements targeted by primary piRNAs, while NcP2 was restricted to the degradation profile. The presence of NcP1.1 and NcP2 was investigated in sandfly populations from South America and the Old World. These EVEs are shared by different sandfly populations in South America while none of the Old World species studied presented the insertions.