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Background Methods Results Conclusions Visceral leishmaniasis (VL) is a severe parasitic disease transmitted by phlebotomine sandflies. VL is endemic in West Pokot County, Kenya, where effective strategies to interrupt transmission are impeded by the limited understanding of VL risk factors. Therefore, this case–control study aimed to explore environmental, behavioural and household determinants of VL in West Pokot.From November 2022 to January 2023, a structured questionnaire was administered to 36 symptomatic primary VL cases attending Kacheliba Sub‐County Hospital in West Pokot and to 50 healthy controls from local villages. The VL status of all participants was confirmed using an rK39 rapid diagnostic test. Associations between questioned determinants and VL were investigated by means of age‐corrected univariate logistic regression analysis.Significant associations were found between VL and housing characteristics, such as window presence and floor type. VL cases more frequently reported the presence of cattle, dogs and sheep in their house yards. VL was also associated with cutting down trees in the house yard and house proximity to several Acacia tree species. Furthermore, outdoor activities, including travelling outside the residence for more than 2 weeks, activities near termite mounds, and forest activities during the rainy season, increased the risk of VL.This work reports a number of previously undescribed risk factors for VL in the understudied West Pokot focus. The results suggest VL transmission occurs both peri‐domestically at night and outdoors during the day, particularly when sandfly resting sites are disturbed. Our findings warrant further research into sandfly ecology and potential zoonotic parasite reservoirs in West Pokot. [ABSTRACT FROM AUTHOR]

Entomological investigations were conducted for the first time in urban forest remnants of Porto Velho, state of Rondônia, Brazil, to explore the transmission dynamics of Leishmania. Sand fly collections were carried out at ten sites, encompassing both canopy and ground strata, from October to December 2021. A total of 1,671 sand flies were collected, representing 42 species within 12 genera. Nyssomyia Antunesi (n = 384) and Psychodopygus davisi (n = 111) were the most abundant species. Molecular analyses targeting the V7V8 region (18S gene) unveiled the presence of sequences 100% identical to Leishmania infantum in females of Bichromomyia flaviscutellata (1), Nyssomyia Antunesi complex (6), Nyssomyia umbratilis (1), Nyssomyia sp. (1), Psychodopygus ayrozai (1), Ps. davisi (3), Psychodopygus paraensis (1), and Sciopemyia sordellii (1). Sequences 100% similar to Trypanosoma minasense were found in two samples of the Nyssomyia Antunesi complex, and two samples of Sc. sordellii presented 100% identity to a Trypanosoma sp. strain, previously identified in this same sand fly in Rondônia. Sequencing of Cytb fragment suggested Homo sapiens, Dasypus novemcinctus and Tamandua tetradactyla as the blood source for distinct sand flies. The identification of sequences similar to L. infantum in sand flies collected in urban forest fragments is noteworthy, correlating with the recent local and regional occurrence of autochthonous cases of human visceral leishmaniasis. However, further studies are imperative to ascertain the presence of hosts/reservoirs and evaluate the risk of L. infantum transmission to humans. [ABSTRACT FROM AUTHOR]

Background: Vector control based on indoor residual spraying (IRS) is one of the main components of the visceral leishmaniasis (VL) elimination programme in India. Dichlorodiphenyltrichloroethane (DDT) was used for IRS until 2015 and was later replaced by the synthetic pyrethroid alpha-cypermethrin. Both classes of insecticides share the same target site, the voltage-gated sodium channel (Vgsc). As high levels of resistance to DDT have been documented in the local sand fly vector, Phlebotomus argentipes, it is possible that mutations in the Vgsc gene could provide resistance to alpha-cypermethrin, affecting current IRS pyrethroid-based vector control. Methods: This study aimed to compare frequencies of knockdown resistance (kdr) mutations in Vgsc between two sprayed and two unsprayed villages in Bihar state, India, which had the highest VL burden of the four endemic states. Across four villages, 350 female P. argentipes were collected as part of a 2019 molecular xenomonitoring study. DNA was extracted and used for sequence analysis of the IIS6 fragment of the Vgsc gene to assess the presence of kdr mutations. Results: Mutations were identified at various positions, most frequently at codon 1014, a common site known to be associated with insecticide resistance in mosquitoes and sand flies. Significant inter-village variation was observed, with sand flies from Dharampur, an unsprayed village, showing a significantly higher proportion of wild-type alleles (55.8%) compared with the three other villages (8.5–14.3%). The allele differences observed across the four villages may result from selection pressure caused by previous exposure to DDT. Conclusions: While DDT resistance has been reported in Bihar, P. argentipes is still susceptible to pyrethroids. However, the presence of kdr mutations in sand flies could present a threat to IRS used for VL control in endemic villages in India. Continuous surveillance of vector bionomics and insecticide resistance, using bioassays and target genotyping, is required to inform India's vector control strategies and to ensure the VL elimination target is reached and sustained. [ABSTRACT FROM AUTHOR]

Introduction Methods Results Conclusions Leishmaniasis stands out as a public health problem in the state of Minas Gerais, Brazil, especially in the Midwest region. However, the entomological aspects in several municipalities remain unknown. Therefore, this study aimed to investigate the sand fly fauna in Bambuí, encompassing ecological dynamics and molecular detection of Leishmania.Monthly collections were conducted using CDC light traps from September 2018 to August 2020 across 16 selected points with urban and rural characteristics, chosen based on the coverage area of the Municipal Health Department and the occurrence of canine and human visceral leishmaniasis (VL) cases. Ecological indices of the sand fly population (Chao1, Shannon, Simpson and Pielou) were assessed, and sand fly abundance was correlated to climatic variables (humidity, temperature and rainfall).A total of 8838 specimens representing 17 species within nine genera were collected (estimated species richness by Chao 1 estimator = 17; SE ± 1.8). Predominantly, Lutzomyia longipalpis, Nyssomyia whitmani and Evandromyia cortelezzii constituted approximately 98% of all captured sand flies. While species richness and diversity displayed variations throughout the study, a positive correlation emerged between temperature (p < 0.0001; r = 0.7767), monthly rainfall (p < 0.0001; r = 0.7810) and sand fly abundance. Molecular analysis revealed Leishmania DNA in 2.05% of female sand flies, with the presence of Leishmania infantum in Lu. longipalpis and both Le. infantum and Leishmania braziliensis in Ev. cortelezzii.The entomological data, coupled with the occurrence of autochthonous cases of canine visceral leishmaniasis, offer valuable insights for evidence‐based strategies to prevent leishmaniasis in Bambuí. [ABSTRACT FROM AUTHOR]

Background: Understanding and mapping the distribution of sandflies and sandfly-associated pathogens (SAPs) is crucial for guiding the surveillance and control effort. However, their distribution and the related risk burden in China remain poorly understood. Methods: We mapped the distribution of sandflies and SAPs using literature data from 1940 to 2022. We also mapped the human visceral leishmaniasis (VL) cases using surveillance data from 2014 to 2018. The ecological drivers of 12 main sandfly species and VL were identified by applying machine learning, and their distribution and risk were predicted in three time periods (2021–2040, 2041–2060, and 2061–2080) under three scenarios of climate and socioeconomic changes. Results: In the mainland of China, a total of 47 sandfly species have been reported, with the main 12 species classified into three clusters according to their ecological niches. Additionally, 6 SAPs have been identified, which include two protozoa, two bacteria, and two viruses. The incidence risk of different VL subtypes was closely associated with the distribution risk of specific vectors. The model predictions also revealed a substantial underestimation of the current sandfly distribution and VL risk. The predicted areas affected by the 12 major species of sandflies and the high-risk areas for VL were found to be 37.9–1121.0% and 136.6% larger, respectively, than the observed range in the areas. The future global changes were projected to decrease the risk of mountain-type zoonotic VL (MT-ZVL), but anthroponotic VL (AVL) and desert-type zoonotic VL (DT-ZVL) could remain stable or slightly increase. Conclusions: Current field observations underestimate the spatial distributions of main sandfly species and VL in China. More active surveillance and field investigations are needed where high risks are predicted, especially in areas where the future risk of VL is projected to remain high or increase. Author summary: Our research provides a comprehensive understanding of the distribution of sandflies and SAPs in China. We have identified 47 sandfly species and 6 SAPs, with 12 main sandfly species forming three distinct ecological clusters. Our machine learning models predict the distribution and risk of sandfly species and VL under various climate and socioeconomic scenarios. The model predictions reveal a significant underestimation of the current sandfly distribution and VL risk. Future global changes are projected to decrease the risk of MT-ZVL, but AVL and DT-ZVL could remain stable or slightly increase. These findings underscore the need for more active surveillance and field investigations of vectors, especially where the future risk of VL is projected to remain high or increase. This study provides important theoretical support for guiding the surveillance and control of sandflies and SAPs. [ABSTRACT FROM AUTHOR]

Abstract Background Visceral 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 . Methods The 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. Results Tree 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. Conclusions The 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.

Background: The sand fly Nyssomyia neivai is one of the most abundant species in Southern Brazil. It is frequently found in areas that are foci of visceral leishmaniasis in the state of Santa Catarina, caused by Leishmania infantum. In this region, the main vector of L. infantum, Lutzomyia longipalpis, has not been detected. In the absence of L. longipalpis, this study aimed to identify the sand fly fauna and diagnose any potential Leishmania spp. infection in sand flies and in dogs in a region of Southern Brazil that experienced a recent canine visceral leishmaniasis outbreak. Methods: This report includes a survey of the sand fly fauna at the Zoonosis Control Center of the Municipality of Tubarão (Santa Catarina, Brazil). Molecular tests were conducted to investigate Leishmania spp. natural infection in sand flies using polymerase chain reaction (PCR). In positive females, in addition to morphological identification, molecular analysis through DNA barcoding was performed to determine the sand fly species. Additionally, the dogs were tested for the presence of Leishmania spp. using a non-invasive technique for the collection of biological material, to be assessed by PCR. Results: A total of 3419 sand flies, belonging to five genera, were collected. Nyssomyia neivai was the most abundant species (85.8%), followed by Migonemyia migonei (13.3%), Pintomyia fischeri (0.8%), Evandromyia edwardsi (< 0.1%), and species of the genus Brumptomyia. (0.1%). Out of the 509 non-engorged females analyzed by PCR, two (0.4%) carried L. infantum DNA. The naturally infected females were identified as Ny. neivai, in both morphological and molecular analysis. In addition, two out of 47 conjunctival swabs from dogs tested positive for L. infantum, yielding an infection rate of 4.2%. Conclusions: These results confirm the presence of Ny. neivai naturally infected with L. infantum in an area where dogs were also infected by the parasite, suggesting its potential role as a vector in Southern Brazil. [ABSTRACT FROM AUTHOR]

In an area endemic with Indian visceral leishmaniasis (VL), we performed direct xenodiagnosis to evaluate the transmission of Leishmania donovani from patients with VL–human immunodeficiency virus (HIV) coinfection to the vector sandflies, Phlebotomus argentipes. Fourteen patients with confirmed VL-HIV coinfection, with a median parasitemia of 42 205 parasite genome/mL of blood, were exposed to 732 laboratory-reared pathogen-free female P argentipes sandflies on their lower arms and legs. Microscopy revealed that 16.66% (122/732) of blood-fed flies were xenodiagnosis positive. Notably, 93% (13/14) of the VL-HIV group infected the flies, as confirmed by quantitative polymerase chain reaction and/or microscopy, and were 3 times more infectious than those who had VL without HIV. [ABSTRACT FROM AUTHOR]

This study investigated the sand fly fauna of the municipality Iguatama, in the Midwest Region of Minas Gerais state, Brazil, including Leishmania infection rates and blood meal sources. Sand flies were collected during four periods over the course of a single year, encompassing both dry and rainy seasons, using CDC light traps placed in peridomiciles where dogs were seropositive for visceral leishmaniasis (VL). A total of 762 sand fly specimens, representing 12 species across seven genera, were collected. Lutzomyia longipalpis was the most abundant species, comprising 57.6% of the collected specimens, followed by Nyssomyia neivai (19.6%) and Nyssomyia whitmani (10.5%). Species richness and diversity varied among collection periods, with the highest diversity observed in January 2019. Molecular analysis detected Leishmania DNA in 12.5% of the sand fly specimens, with Le. infantum being the predominant species. Blood meal analysis revealed feeding on multiple vertebrate species, including humans, rats, dogs, and chickens. The presence of Leishmania DNA in sand flies, and the identification of human blood meals, highlight the potential role of these species in VL transmission. These findings underscore the importance of continued surveillance and control measures to prevent the spread of VL and reduce transmission risk in the region. [ABSTRACT FROM AUTHOR]

In Nepal, visceral leishmaniasis (VL) has been targeted for elimination as a public health problem by 2026. Recently, increasing numbers of VL cases have been reported from districts of doubtful endemicity including hills and mountains, threatening the ongoing VL elimination program in Nepal. We conducted a multi-disciplinary, descriptive cross-sectional survey to assess the local transmission of Leishmania donovani in seven such districts situated at altitudes of up to 1,764 meters in western Nepal from March to December 2019. House-to-house surveys were performed for socio-demographic data and data on past and current VL cases. Venous blood was collected from all consenting individuals aged ≥2 years and tested with the rK39 RDT. Blood samples were also tested with direct agglutination test, and a titer of ≥1:1600 was taken as a marker of infection. A Leishmania donovani species-specific PCR (SSU-rDNA) was performed for parasite species confirmation. We also captured sand flies using CDC light traps and mouth aspirators. The house-to-house surveys documented 28 past and six new VL cases of which 82% (28/34) were without travel exposure. Overall, 4.1% (54/1320) of healthy participants tested positive for L. donovani on at least one serological or molecular test. Among asymptomatic individuals, 17% (9/54) were household contacts of past VL cases, compared to 0.5% (6/1266) among non-infected individuals. Phlebotomus argentipes, the vector of L. donovani, was found in all districts except in Bajura. L. donovani was confirmed in two asymptomatic individuals and one pool of sand flies of Phlebotomus (Adlerius) sp. We found epidemiological and entomological evidence for local transmission of L. donovani in areas previously considered as non-endemic for VL. The national VL elimination program should revise the endemicity status of these districts and extend surveillance and control activities to curb further transmission of the disease. [ABSTRACT FROM AUTHOR]

Leishmaniasis refers to a disease with a wide range of manifestations; and there are three main forms of disease, cutaneous, mucocutaneous, and visceral. Leishmaniasis is one of the diseases with a protozoan agent which is vector-borne. Visceral leishmaniasis (VL) is the most severe form that can be fiercely life-threatening if left untreated. VL can be caused by members of Leishmania donovani complex, in Iran, Leishmania infantum is considered the primary causative agent of VL, resulting in a zoonotic form of VL. The two main goals of our work, which followed our prior sero-epidemiological and entomological survey, were to characterize and conduct a phylogenetic analysis of the Leishmania species that infect people, dogs, and sandflies. The samples were collected throughout 2017, from January to December, so blood samples were collected from humans and dogs, while sandfly samples were collected with sticky traps. DNA extracted from all seropositive samples of humans and dogs, 10% of sero-negative human samples, and all collected sandflies were subjected to kDNA-nested-PCR for tracing parasites. A total of 30 samples, including 20 human samples, 8 dog samples, and 2 sandfly samples, were found positive for the kDNA gene of L. infantum. Sequences were evaluated to study the genetic diversity among the six discovered L. infantum. Based on kDNA, the phylogenetic study of L. infantum demonstrated a high level of genetic variety and a relationship between the host, the parasite's geographic origin, and its genetic diversity. [ABSTRACT FROM AUTHOR]

Background: 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. Methods: 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. Results: 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. Conclusion: 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. [ABSTRACT FROM AUTHOR]

Phlebotomine sand flies (Diptera: Phlebotominae) are vectors of human and animal pathogens, including Leishmania species protozoan parasites and viruses of the genus Phlebovirus. In Europe, visceral zoonotic leishmaniasis caused by Leishmania infantum , a deadly disease when left untreated, is endemic in southern countries, and dogs are the main reservoir hosts for human infection. Most phleboviruses cause asymptomatic infections or flu-like syndromes in humans, but Toscana phlebovirus can cause meningitis and encephalitis. These diseases are likely to re-emerge, posing a growing threat to public and animal health. Potential triggers include the movement of humans and dogs, increasing numbers of immunosuppressive conditions, climate change and other human-mediated environmental changes. An overview of the main epidemiological characteristics of the pathogens transmitted by sand flies in Europe and the potential triggers involved in their emergence and re-emergence are reviewed here. There is a need to implement mandatory notification of human and canine leishmaniases and human phleboviruses and coordinated epidemiological surveillance programmes at a European level, and to raise awareness among healthcare professionals and citizens about sand fly-borne diseases, following a One Health approach. [Display omitted] [ABSTRACT FROM AUTHOR]

Background: In Brazil, human visceral leishmaniasis (HVL) is caused by the protozoan parasite Leishmania infantum, primarily transmitted by the sand fly Lutzomyia longipalpis, with dogs acting as the main urban reservoir. This study aims to evaluate the effectiveness of 4% deltamethrin-impregnated dog collars (DMC) on HVL incidence. Methods: This is a community intervention study carried out from 2012 to 2015 in the municipalities of Araguaína, State of Tocantins, and Montes Claros, State of Minas Gerais, Brazil. Two areas in each were randomly allocated to either (1) culling seropositive dogs + residual insecticide spraying (control area—CA) or (2) culling seropositive dogs + residual insecticide spraying + DMC fitted to dogs every six months for two years (intervention area—IA). Cases of HVL (n = 1202) occurring from 2008 to 2020 were identified from the Brazilian Reportable Diseases Information System and georeferenced to the control and intervention areas. The HVL cases from 2008 to 2012 were considered as occurring in the "pre-intervention" period. Those cases from 2013 to 2016 and from 2017 to 2020 were regarded as occurring in the "intervention" and "post-intervention" periods, respectively. We used a mixed-effects Poisson regression model to estimate the effectiveness of the intervention, comparing the changes from the pre-intervention period to the intervention and post-intervention periods in the control and intervention areas. Results: In Araguaína, there was a statistically significant reduction in the incidence of HVL in both the control and intervention areas, comparing both the intervention and post-intervention periods with the pre-intervention period. The intervention with DMC was significantly associated with a reduction in HVL when comparing the intervention and pre-intervention periods, yielding an effectiveness estimate of the DMC of 27% (IC95% 1–46%, p = 0.045). No differences were observed when comparing the pre- and post-intervention periods (p = 0.827). In Montes Claros, cases reduced in both the control and intervention areas from the pre-intervention period to the intervention period (p = 0.913). In the post-intervention period, the incidence increased in the control area, while cases continued to decrease in the DMC area (p = 0.188). Conclusions: The use of DMC was associated with a reduction of 27% in the incidence of HVL during the period of DMC delivery, indicating that DMC is effective as an additional strategy for controlling visceral leishmaniasis in Brazil. However, no significant reduction associated with DMC was detected after the intervention period, suggesting that a control program based on the large-scale deployment of DMC might have to be maintained for more extended periods without interruption. [ABSTRACT FROM AUTHOR]

Background: 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. Methods: 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. Results: 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 Central Asian countries such as Afghanistan, Iran, Lebanon, Turkey, and Georgia. In Iran, the first infection of Phlebotomus kandelakii and Ph. perfiliewi tranccaucasicus 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 and Crimea), Ph. syriacus (Southwest Asia, Caucasus), Ph. notus (Afghanistan), Ph. wenyoni (Iran and Iraq), and Ph.wui (China). The first natural infection of Phlebotomus alexandri with Leishmania infantum was reported in 2006 in endemic foci of visceral leishmaniasis in Fars province, southern Iran. Conclusion: 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 .

Background: Leishmaniasis is a common neglected tropical disease in Ethiopia. Visceral leishmaniasis (VL) caused by Leishmania donovani presents in the lowlands, while cutaneous leishmaniasis (CL) affects people living in the highlands. Although CL is described as being caused by Leishmania aethiopica, there is also evidence of L. tropica and L. major isolated from a patient, sand flies and potential reservoirs. Information on species causing CL in Ethiopia is patchy, and no nation-wide study has ever been done. Understanding which species are causing CL in Ethiopia can have important implications for patient management and disease prevention. Methods: We analyzed stored routine samples and biobanked DNA isolates from previously conducted studies of CL patients from different centers in the north, center and south of Ethiopia. Species typing was performed using ITS-1 PCR with high-resolution melt (HRM) analysis, followed by HSP70 amplicon sequencing on a selection of the samples. Additionally, sociodemographic, clinical and laboratory data of patients were analyzed. Results: Of the 226 CL samples collected, the Leishmania species could be determined for 105 (45.5%). Leishmania aethiopica was identified in 101 (96.2%) samples from across the country. In four samples originating from Amhara region, northwestern Ethiopia, L. donovani was identified by ITS-1 HRM PCR, of which two were confirmed with HSP70 sequences. While none of these four patients had symptoms of VL, two originated from known VL endemic areas. Conclusions: The majority of CL was caused by L. aethiopica, but CL due to L. tropica and L. major cannot be ruled out. Our study is the first to our knowledge to demonstrate CL patients caused by L. donovani in Ethiopia. This should spark future research to investigate where, how and to which extent such transmission takes place, how it differs genetically from L. donovani causing VL and whether such patients can be diagnosed and treated successfully with the currently available tools and drugs. [ABSTRACT FROM AUTHOR]

Miranda Municipality of Mato Grosso do Sul, borders the Pantanal wetland, a famous fishing destination visited by tourists from all over the world, and is a location where visceral leishmaniasis has been reported. To assess the risk of Leishmania infantum transmission, we studied the sandfly community, focusing on known vector and parasite presence. We conducted light trap collections twice per month at nine sites within the city (including two forested areas) for one year. We collected a total of 12,727 sand flies, 10,891 males and 1,836 females belonging to 11 species: Brumptomyia avellari, Evandromyia aldafalcaoae, Ev. evandroi, Ev. lenti, Ev. sallesi, Ev. walkeri, Lu. longipalpis, Nyssomyia whitmani, Psathyromyia bigeniculata, Pa. hermanlenti and Pa. punctigeniculata. Lutzomyia longipalpis, the proven vector of Leishmania infantum, was captured each month, and was the most abundant species observed, accounting for more than 99% of sand flies captured in most sites, especially where chicken coops were present. Evidence of Leishmania infantum infection was detected in 0.40% of Lu. longipalpis tested. We developed a generalized mixed multilevel model for Lu. longipalpis, that includes within-year seasonality, location of capture (indoors vs. outdoors), vector abundance, and sex ratio. The VL vector was abundant both inside and outside houses. Large numbers of Lu. longipalpis were observed in outdoor sites where domestic animals were present but were absent from forest sites. Our findings suggest high vector populations and Le. infantum presence in a city where tourists could be exposed to visceral leishmaniasis, with significant implications for more surveillance and control activities. Author summary: Several localities in Mato Grosso do Sul have already recorded human visceral leishmaniasis cases, including the city of Miranda, which is a famous fishing and tourism destination. Due to the high number of visitors, we decided to evaluate the risk of Leishmania infantum transmission in the area. Thus, we conducted vector collections over one year, detecting both the presence of the main vector (Lutzomyia longipalpis) and the Le. infantum parasite. We used nonlinear multilevel mixed models, under the Bayesian framework, to investigate the seasonal changes on transmission risk (i.e.; overall vector abundance and the proportion of females), taking into account the residence space. We found that the overall vector abundance outdoors was higher than in the indoors. Outdoors abundance was more consistent with two-peaks during the year, whereas indoors abundance was more seasonal with a single peak. Although, indoor abundance was lower than observed outdoors, the proportion of females captured was up to twice that observed outdoors. These findings highlighted the spatiotemporal component of leishmaniasis transmission risk, which can guide entomological surveillance and control measures. [ABSTRACT FROM AUTHOR]

Visceral leishmaniasis (VL, kala azar), caused by Leishmania donovani, transmitted by Phlebotomus orientalis, is a serious systemic disease that causes high morbidity and mortality rates in Sudan and other parts of East Africa and the world. Despite progress in understanding the epidemiology of the disease in East Africa, little is known about the host preference of P. orientalis in kala azar endemic villages of Sudan, which have some of the highest VL incidence rates in the world. The present study used host choice experiments and blood‐meal identification approaches to determine the host preference of P. orientalis in kala azar endemic villages in Gedarif state, eastern Sudan. In the host choice experiment, tent traps were used to compare the attractiveness of cows, donkeys, sheep and goats for host‐seeking P. orientalis. In the blood‐meal identification study, blood‐fed P. orientalis females, captured inside houses and peri‐domestic habitats, were subjected to molecular typing using cytochrome b gene (cyt b) amplification and sequence analysis. Cows and donkeys were the most attractive to blood‐seeking P. orientalis, followed by goats. Similarly, the blood‐meal analysis of P. orientalis showed that the vector preferentially feeds on cows, followed by donkeys, humans and goats. The human blood index of P. orientalis was 19.4% (42/216), indicating a high zoophilic habit of the vector, both inside and outside the houses. Although the order of host preference varied by location, it was clear that cows are the most preferred host of P. orientalis in the area. Results are discussed in relation to the role of domestic/livestock animals in VL zoopotentiation and zooprophylaxis. Inference is made on the potential impact of insecticide treatment of cows in control of the vector and the transmission of VL in Sudan and other parts of East Africa. [ABSTRACT FROM AUTHOR]

Background: Visceral leishmaniasis (VL), a life-threatening neglected tropical disease, is targeted for elimination from Nepal by the year 2026. The national VL elimination program is still confronted with many challenges including the increasingly widespread distribution of the disease over the country, local resurgence and the questionable efficacy of the key vector control activities. In this study, we assessed the status and risk of Leishmania donovani transmission based on entomological indicators including seasonality, natural Leishmania infection rate and feeding behavior of vector sand flies, Phlebotomus argentipes, in three districts that had received disease control interventions in the past several years in the context of the disease elimination effort. Methods: We selected two epidemiologically contrasting settings in each survey district, one village with and one without reported VL cases in recent years. Adult sand flies were collected using CDC light traps and mouth aspirators in each village for 12 consecutive months from July 2017 to June 2018. Leishmania infection was assessed in gravid sand flies targeting the small-subunit ribosomal RNA gene of the parasite (SSU-rRNA) and further sequenced for species identification. A segment (~ 350 bp) of the vertebrate cytochrome b (cytb) gene was amplified from blood-fed P. argentipes from dwellings shared by both humans and cattle and sequenced to identify the preferred host. Results: Vector abundance varied among districts and village types and peaks were observed in June, July and September to November. The estimated Leishmania infection rate in vector sand flies was 2.2% (1.1%–3.7% at 95% credible interval) and 0.6% (0.2%–1.3% at 95% credible interval) in VL and non-VL villages respectively. The common source of blood meal was humans in both VL (52.7%) and non-VL (74.2%) villages followed by cattle. Conclusions: Our findings highlight the risk of ongoing L. donovani transmission not only in villages with VL cases but also in villages not reporting the presence of the disease over the past several years within the districts having disease elimination efforts, emphasize the remaining threats of VL re-emergence and inform the national program for critical evaluation of disease elimination strategies in Nepal. [ABSTRACT FROM AUTHOR]

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

Background: The kala-azar elimination programme has resulted in a significant reduction in visceral leishmaniasis (VL) cases across the Indian Subcontinent. To detect any resurgence of transmission, a sensitive cost-effective surveillance system is required. Molecular xenomonitoring (MX), detection of pathogen DNA/RNA in vectors, provides a proxy of human infection in the lymphatic filariasis elimination programme. To determine whether MX can be used for VL surveillance in a low transmission setting, large numbers of the sand fly vector Phlebotomus argentipes are required. This study will determine the best method for capturing P. argentipes females for MX. Methodology/Principal findings: The field study was performed in two programmatic and two non-programmatic villages in Bihar, India. A total of 48 households (12/village) were recruited. Centers for Disease Control and Prevention light traps (CDC-LTs) were compared with Improved Prokopack (PKP) and mechanical vacuum aspirators (MVA) using standardised methods. Four 12x12 Latin squares, 576 collections, were attempted (12/house, 144/village,192/method). Molecular analyses of collections were conducted to confirm identification of P. argentipes and to detect human and Leishmania DNA. Operational factors, such as time burden, acceptance to householders and RNA preservation, were also considered. A total of 562 collections (97.7%) were completed with 6,809 sand flies captured. Females comprised 49.0% of captures, of which 1,934 (57.9%) were identified as P. argentipes. CDC-LTs collected 4.04 times more P. argentipes females than MVA and 3.62 times more than PKP (p<0.0001 for each). Of 21,735 mosquitoes in the same collections, no significant differences between collection methods were observed. CDC-LTs took less time to install and collect than to perform aspirations and their greater yield compensated for increased sorting time. No significant differences in Leishmania RNA detection and quantitation between methods were observed in experimentally infected sand flies maintained in conditions simulating field conditions. CDC-LTs were favoured by householders. Conclusions/Significance: CDC-LTs are the most useful collection tool of those tested for MX surveillance since they collected higher numbers of P. argentipes females without compromising mosquito captures or the preservation of RNA. However, capture rates are still low. Author summary: Molecular xenomonitoring, screening insects for pathogen DNA/RNA, may be used for surveillance of diseases transmitted by insects. Since the proportion of insects infected with pathogens is very low in areas targeted for disease elimination, large numbers of females need to be screened. We compared three different methods for collecting Phlebotomus argentipes sand fly females, the vector of parasites causing the disease visceral leishmaniasis in the Indian subcontinent, to determine which collected the largest number of females. Other factors that may also influence selection of a particular method of collection by a disease control programme, such as the time it takes to collect and sort samples, the acceptance of householders for a particular collection method and whether RNA degradation in insect samples differed between collection methods, were also considered. Centers for Disease Control and Prevention light traps (CDC-LTs) proved to be more useful than two types of aspiration methods for collecting higher numbers of sand fly females and RNA preservation was retained. Furthermore, they took less time to install than to perform aspirations and were favoured by householders. Therefore, CDC-LTs were considered to be the most suitable collection method for molecular xenomonitoring of sand flies in India. [ABSTRACT FROM AUTHOR]

Purpose: In Turkey, the main causative agent of visceral leishmaniasis (VL) is Leishmania. infantum and the main causative agent of cutaneous leishmaniasis (CL) is Leishmania tropica. In this study, we aimed to discuss the possible mechanisms, clinical aspects, and threat of visceralizing L. tropica. Methods: This study includes seven cases of VL caused by L. tropica.Five patients were male (71%) and four were adults (57%). Results: All the VL patients complained of fever and splenomegaly. Fatigue, pancytopenia, and hepatomegaly were present in six patients each (86%), while weight loss and gastrointestinal system (GIS) symptoms were present in 5 patients (71%). Conclusions: In this study, we have evaluated seven cases of visceralized L. tropica (VLT) in the context of the changing leishmaniasis epidemiology in Turkey. We have evaluated the possible mechanisms of visceralization; inter- and intraspecies genetic exchange with all the old world leishmaniasis agents present in the region, stress induced by inappropriate use of drugs, and possible ongoing adaptation mechanisms of Leishmania spp. The threat posed by VLT is significant as L. tropica is the most widespread and most common cause of leishmaniasis in Turkey. We do not know the vectorial capacity of the sand flies for the transmission of VLT strains or if these strains are in circulation in Turkey. Future studies should be carried out to investigate these issues as the transition of L. tropica from a mild disease-causing agent to a mortal one poses a significant public health concern for Turkey and Europe. [ABSTRACT FROM AUTHOR]

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

We consider the transmission of visceral leishmaniasis in three populations: humans, dogs and sandflies. To investigate the effects of spatial heterogeneity and temporal periodicity on disease transmission, a nonlocal periodic reaction‐diffusion equation model is developed. We introduce the basic reproduction number R0$$ {\mathcal{R}}_0 $$ and establish the global dynamics. Numerical simulations show that (i) R0$$ {\mathcal{R}}_0 $$ is overestimated in the absence of spatial heterogeneity; (ii) R0$$ {\mathcal{R}}_0 $$ is underestimated without considering the effect of temporal seasonality; and (iii) shortening periodic delays of visceral leishmaniasis reduces the risk of disease transmission. [ABSTRACT FROM AUTHOR]

Background: In the Somali region of Ethiopia, visceral leishmaniasis (VL) is a public health concern. However, VL epidemiology and sand fly vectors have not been well studied in various areas of the regional state, including Denan district. Therefore, this study was conducted to determine the sero-prevalence, associated factors, and distribution of sand fly vectors of VL in Denan district, south-eastern Ethiopia. Methods: A facility-based cross-sectional study was conducted from April to September 2021 among VL patients with classic signs and symptoms visiting Denan Health Center in south-eastern Ethiopia. Using a convenience sampling method, 187 blood samples were collected from individuals who visited Denan Health Center during the study period. Blood samples were subjected to Direct Agglutination Test for the detection of antibodies to VL. A pre-tested structured questionnaire was also used to gather information on risk factors and other characteristics of knowledge and attitude assessment. Sand flies were also collected from indoor, peri-domestic, mixed forest, and termite mounds using light and sticky traps to determine the fauna and abundance. Results: The overall sero-prevalence rate was 9.63% (18/187). The sero-prevalence was significantly associated with outdoor sleeping (OR = 2.82), the presence of damp floors (OR = 7.76), and sleeping outdoor near animals (OR = 3.22). Around 53.48% of the study participants had previously heard about VL. Study participants practiced different VL control methods, including bed nets (42%), insecticide spraying (32%), smoking plant parts (14%), and environmental cleaning (8%). In total, 823 sand fly specimens, comprising 12 species in two genera (Phlebotomus and Sergentomyia), were trapped and identified. The most abundant species was Sergentomyia clydei (50.18%), followed by Phlebotomus orientalis (11.42%). Also, a higher proportion of P. orientalis was found in termite mounds (65.43%), followed by mixed forest (37.8%) and peri-domestic (20.83%) habitats. Conclusion: The study demonstrated a 9.63% sero-positivity of VL and a remarkable gap in knowledge, attitude, and practices towards VL. P. orientalis was also detected, which could be a probable vector in this area. Thus, public education should be prioritized to improve the community's awareness of VL and its public health impact. In addition, detailed epidemiological and entomological studies are recommended. [ABSTRACT FROM AUTHOR]

A study was carried out to compare the infection rates of Leishmania donovani in Phlebotomus orientalis sandflies at different microhabitats of a VL endemic village in Gedarif state, Sudan. DNA extracts of 1078 P. orientalis sand fly females sampled by CDC light traps from indoor, outdoor, peri-domestic, and sylvatic sites, in three transmission seasons, March–June 2016–18, in Helat-Belo village, were subjected to independent PCR amplifications targeting Leishmania kDNA and the cpb gene followed by ITS1 region sequencing. Leishmania kDNA was detected in 1.4% of the 1078 P. orientalis females captured in the area. Two of these specimens showed a characteristic 741 bp band of L. donovani after cpb gene amplification. The DNA sequence of the ITS1 region of the parasites matched the ITS1 L. donovani genotype F. There were no signficant differences between rates of infection of L. donovani in P. orientalis captured at different sites. Blood meals found in infected flies origninated from human (5 specimens), cattle (4 specimens) and donkey (2 specimens). The finding of fresh cow and donkey blood in the infected flies suggests the possible role of these animals in the zoopotentiation and/or zooprophylaxis against VL. The study provides important information for VL transmission models and control programs in East Africa.

Visceral leishmaniasis (VL) is a vector-borne protozoan disease, which can be fatal if left untreated. Synthetic chemical insecticides are very effective tools for controlling of insect vectors, including the sand fly Phlebotomus argentipes, the vector of VL in the Indian subcontinent. However, repeated use of the same insecticide with increasing doses potentially can create high selection pressure and lead to tolerance and resistance development. The objective of this study was to determine the lethal concentrations and assess levels of susceptibility, diagnostic doses and times to death of laboratory-reared P. argentipes to five insecticides that are used worldwide to control vectors. Using the Center for Disease Control and Prevention (CDC) bottle bioassay, 20–30 sand flies were exposed in insecticide- coated 500-ml glass bottles. Flies were then observed for 24 hours and mortality was recorded. Dose-response survival curves were generated for each insecticide using QCal software and lethal concentrations causing 50%, 90% and 95% mortality were determined. A bioassay was also conducted to determine diagnostic doses and diagnostic times by exposing 20–30 flies in each bottle containing set concentrations of insecticide. Mortality was recorded at 10-minute intervals for 120 minutes to generate the survival curve. Phlebotomus argentipes are highly susceptible to alpha-cypermethrin, followed by deltamethrin, malathion, chlorpyrifos, and least susceptible to DDT. Also, the lowest diagnostic doses and diagnostic times were established for alpha-cypermethrin (3μg/ml for 40 minutes) to kill 100% of the flies. The susceptibility data, diagnostic doses and diagnostic times presented here will be useful as baseline reference points for future studies to assess insecticide susceptibility and resistance monitoring of field caught sand flies and to assist in surveillance as VL elimination is achieved in the region. Author summary: Synthetic chemical insecticides are mainly used for controlling sand flies via indoor residual spraying (IRS) and insecticide treated nets. Sand flies had developed resistance against DDT in endemic areas of Bihar, India, and was replaced by synthetic pyrethroids (alpha-cypermethrin) in the second phase of IRS in 2015. The substantial increase in the use of synthetic pyrethroids will potentially lead to increased resistance because DDT and pyrethroids both have the same target site. To monitor insecticide resistance in field caught sand flies, susceptibility status and diagnostic doses and times are warranted, as no diagnostic dose for resistance detection has been established for Indian sand flies. A laboratory reared P. argentipes was used to quantify insecticide susceptibility and determination of diagnostic doses and times. The CDC bottle bioassay was used with five insecticides viz., alpha-cypermethrin, deltamethrin, malathion, chlorpyrifos and DDT. Results indicate that the P. argentipes were highly susceptible to alpha-cypermethrin followed by deltamethrin, malathion, chlorpyrifos and least susceptible to DDT. This study provides critical base line reference data for further studies to assess insecticide susceptibility and resistance monitoring in field populations, as well as determining diagnostic doses and times for other insecticide susceptible sand fly populations undergoing surveillance to sustain VL elimination. [ABSTRACT FROM AUTHOR]

Cutaneous leishmaniasis caused by Leishmania major or L. tropica and visceral leishmaniasis caused by L. infantum have been reported in Israel. We collected Phlebotomus spp. sand flies in the Negev desert of southern Israel to identify circulating Leishmania spp. Of 22,636 trapped sand flies, 80% were P. alexandri. We sequenced Leishmania-specific internal transcribed spacer 1 fragments and K26 genes. Of 5,019 Phlebotomus female sand flies, 2.5% were Leishmania DNA–positive; 92% of infections were L. donovani. Phylogenetic analyses showed separate clustering of L. donovani and L. infantum. P. alexandri flies positive for L. donovani harbored blood meals from European hares. Leishmania DNA isolated from a patient with cutaneous leishmaniasis who lived in the survey area was identical to L. donovani from P. alexandri flies. We report circulation of L. donovani, a cause of visceral leishmaniasis, in southern Israel. Prompt diagnosis and Leishmania spp. identification are critical to prevent leishmaniasis progression. [ABSTRACT FROM AUTHOR]

Sand flies are the exclusive vectors of leishmaniasis. This group of parasitic diseases is a serious public health problem in Morocco. The aim of this study was to investigate the sand fly fauna, mainly the species composition, biodiversity, and seasonal activity of sand flies in El Hajeb in central Morocco. A total of six stations (Aït Naaman, Aït Rbaa, Aït Brahim, Ain Taoujdate, Sidi Mbarek, and Aït Oufella) were studied, five of which had recently recorded cases of leishmaniasis. Sand fly bimonthly captures were carried out using a sticky paper trap in different biotopes from March to December 2019. A total of 14590 adult sand flies were collected. The activity of the sand fly started in April and declined in November. The periods of high abundance were July, September, and November. Morphological identification of sand flies shows the presence of twelve species: Phlebotomus papatasi, P. longicuspis, P. perniciosus, P. sergenti, P. bergeroti, P. alexandri, P. dreyfussi, P. ariasi, Sergentomyia fallax, S. minuta, S. schwetzi, and S. antennata. The analysis showed that species belonging to the genus Phlebotomus were the most dominant (93.3%) and the risk periods were spread during the summer and autumn seasons. The present study provides for the first time information on the species of sand flies in El Hajeb. It, therefore, provides decision makers with an important tool to conduct vector control actions during peak periods in order to limit the transmission of leishmaniasis. A preprint was made available by the research square in the following link: "https://assets.researchsquare.com/files/rs-1409330/v1/dfef7013-0327-4a54-897f-214924a2d950.pdf?c=1646838874." [ABSTRACT FROM AUTHOR]

The continuously expanding distribution of sand flies, proven vectors of Leishmania and of several phleboviruses, is a growing public health issue in Europe. Especially in Italy, visceral leishmaniasis (VL) is occurring with increasing incidence northward, in previously non-endemic provinces. Around the globe, disease elimination efforts largely focus on sand fly vector insecticidal control, often leading to the development of resistance. In Emilia-Romagna (ER), northern Italy, insecticides are heavily applied for agricultural and mosquito control, but not specifically against sand flies. Here, we investigated the sand fly species composition in certain environmental settings in ER provinces and monitored the presence of pyrethroid resistance mutations and pathogen circulation. Phlebotomus perfiliewi, a dominant vector of Leishmania infantum, was detected almost exclusively in the region. No mutations in the voltage-gated sodium channel gene, e.g., knock-down resistance mutations I1011M, L1014F/S, V1016G, or F1020S, were recorded. Pathogen monitoring revealed that almost 40% of the tested sand fly pools were positive for Leishmania, while the presence of Toscana and Fermo phleboviruses was also observed in much lower frequencies (≤3% positive pools). Regular epidemiological and entomological monitoring, alongside resistance surveillance, is highly recommended to ensure the sustainability and efficiency of vector control interventions. [ABSTRACT FROM AUTHOR]

Abstract Background In the State of Mato Grosso do Sul, Brazil, sand flies and cases of visceral (VL) and cutaneous (CL) leishmaniases have been reported in almost all municipalities. The aim of this study was to analyze the geographic distribution of VL and CL in relation the sand fly species found in the municipalities of Mato Grosso do Sul. Methods We analyzed VL and CL cases from 2001 to 2018 using data from the Notifiable Diseases Information System (SINAN). Data collected since 2003 on the presence of sand fly vectors (proven or suspected) were provided by the State Health Secretariat. Results A total of 3566 and 3030 cases of VL and CL, respectively, were reported from 2001 to 2018. The municipalities with the most reported cases of VL were Campo Grande (2495), Três Lagoas (442), Corumbá (140) and Aquidauana (136); and those for CL were Campo Grande (635) and Bodoquena (197). The following sand fly species with vector potential were found in 59 municipalities (74.7%): Lutzomyia longipalpis, Lutzomyia cruzi, Nyssomyia whitmani, Migonemyia migonei, Nyssomyia neivai, Pintomyia pessoai, Bichromomyia flaviscutellata and Pintomyia fischeri. Sand flies were present in six municipalities where no cases of VL were reported and in two municipalities where no cases of CL were reported. Conclusions Our results indicate that the geographical distribution of VL and CL in Mato Grosso do Sul expanded during the study period, and highlight the presence of sand fly vectors in municipalities where these diseases are currently considered to be non-endemic. Graphical Abstract

Visceral and cutaneous leishmaniasis are endemic to specific regions due to the ecological preferences of phlebotomine sand flies and Leishmania spp. transmission. Sand fly entomological data in northern Kenya are scarce due to limited studies and neglect of leishmaniasis. The aim of this study was to investigate: (i) sand fly diversity and distribution; (ii) occurrence of Leishmania DNA within sand flies; and (iii) blood-meal sources of sand flies in Laisamis, northern Kenya. We conducted an entomological survey during February and March of 2021 in five areas of Laisamis sub-county using standard CDC light traps. A total of 1009 sand flies (394 male and 615 female) were morphologically identified, and representative samples verified by PCR amplification and sequencing of the cytochrome c oxidase subunit 1 (cox1) gene. Similarly, we identified blood-meal sources and Leishmania DNA in female sand flies by PCR amplicon sequencing of the vertebrate cytochrome b (cyt b) gene and internal transcribed spacer 1 (ITS1) of the 28S rRNA gene, respectively. Sergentomyia clydei (59.8%) was the most abundant sand fly species. Though collected mainly from one locality (Tirgamo), 14.8% of samples belonged to Phlebotomus (Artemievus) alexandri Sinton, 1928. We detected DNA of Leishmania major in 5.19% of Ph. alexandri, whereas Leishmania adleri DNA was detected in S. clydei (7.51%), Sergentomyia squamipleuris (8.00%), and Sergentomyia africanus (8.33%). Nine of 13 blood-fed sand flies had obtained blood from humans, of which 33.3% had L. major DNA. Both Ph. alexandri and S. clydei primarily fed on humans and could potentially be involved in the transmission of cutaneous leishmaniasis. The findings of this study contribute to the understanding of sand fly vector populations and their potential to transmit leishmaniasis in the area.

Background: Visceral leishmaniasis (VL) is one of the most important neglected tropical diseases. Although VL was controlled in several regions of China during the last century, the mountain-type zoonotic visceral leishmaniasis (MT-ZVL) has reemerged in the hilly areas of China in recent decades. The purpose of this study was to construct an indicator framework for assessing the risk of the MT-ZVL in China, and to provide guidance for preventing disease. Methods: Based on a literature review and expert interview, a 3-level indicator framework was initially established in November 2021, and 28 experts were selected to perform two rounds of consultation using the Delphi method. The comprehensive weight of the tertiary indicators was determined by the Delphi and the entropy weight methods. Results: Two rounds of Delphi consultation were conducted. Four primary indicators, 11 secondary indicators, and 35 tertiary indicators were identified. The Delphi-entropy weight method was performed to calculate the comprehensive weight of the tertiary indicators. The normalized weights of the primary indicators were 0.268, 0.261, 0.242, and 0.229, respectively, for biological factors, interventions, environmental factors, and social factors. The normalized weights of the top four secondary indicators were 0.122, 0.120, 0.098, and 0.096, respectively, for climatic features, geographical features, sandflies, and dogs. Among the tertiary indicators, the top four normalized comprehensive weights were the population density of sandflies (0.076), topography (0.057), the population density of dogs, including tethering (0.056), and use of bed nets or other protective measures (0.056). Conclusions: An indicator framework of transmission risk assessment for MT-ZVL was established using the Delphi-entropy weight method. The framework provides a practical tool to evaluate transmission risk in endemic areas. [ABSTRACT FROM AUTHOR]

Leishmaniasis is one of the top neglected tropical diseases with significant morbidity and mortality in low and middle-income countries (LMIC). However, this disease is also spreading in the developed world. Currently, there is a lack of effective strategies to control this disease. Vaccination can be an effective measure to control leishmaniasis and has the potential to achieve disease elimination. Recently, we have generated centrin gene-deleted new world L. mexicana (LmexCen−/−) parasites using CRISPR/Cas9 and showed that they protect mice against a homologous L. mexicana infection that causes cutaneous disease. In this study, we tested whether LmexCen−/− parasites can also protect against visceral leishmaniasis caused by L. donovani in a hamster model. We showed that immunization with LmexCen−/− parasites is safe and does not cause lesions. Furthermore, such immunization conferred protection against visceral leishmaniasis caused by a needle-initiated L. donovani challenge, as indicated by a significant reduction in the parasite burdens in the spleen and liver as well as reduced mortality. Similar control of parasite burden was also observed against a sand fly mediated L. donovani challenge. Importantly, immunization with LmexCen−/− down-regulated the disease promoting cytokines IL-10 and IL-4 and increased pro-inflammatory cytokine IFN-γ resulting in higher IFN-γ/IL-10 and IFN-γ/IL4 ratios compared to non-immunized animals. LmexCen−/− immunization also resulted in long-lasting protection against L. donovani infection. Taken together, our study demonstrates that immunization with LmexCen−/− parasites is safe and efficacious against the Old World visceral leishmaniasis. [ABSTRACT FROM AUTHOR]

Simple Summary: Sand flies are small dipterans of medical importance, as some species are vectors of American visceral leishmaniasis disease (AVL). To determine whether the community of these vectors in forest areas and built areas are influenced by environmental conditions in the city of João Pessoa (state of Paraíba, Brazil), collections were carried out between March 2019 and July 2021. The results showed that forested areas have more non-ALV vector species; however, in the built areas, the predominant species was Lutzomyia longipalpis, a known vector of AVL. This suggests that L. longipalpis adapts better to anthropized areas than to more preserved areas. Thus, health authorities must intensify AVL control programs in the locality. Sand flies are dipterans of medical importance, as some species are vectors of American visceral leishmaniasis (AVL). The municipality of João Pessoa (Paraíba, northeastern Brazil), is an endemic region for AVL, having high rates of human and canine cases. The main objective was to evaluate the sand fly fauna among forest fragments and built areas, and its relationship with environmental conditions. HP light traps were placed in the studied areas from March 2019 to July 2021. A total of 2141 specimens of phlebotomines were captured, comprising nine genera and ten species. Temperature and humidity were significant and positive only in built areas. The diversity composition among forest fragments and built areas was different and the AVL vector, Lutzomyia longipalpis, was the most prevalent species in built areas. The study showed that the built areas present differences in their richness and diversity of sand flies in relation to forest fragments, concluding that the conservation of forest areas, even if urban fragments, favors the diversity of phlebotomine species. [ABSTRACT FROM AUTHOR]

[Display omitted] • Leishmania infantum is the only Leishmania species circulating in Italy. • In the Emilia-Romagna region, humans and dogs are infected by two distinct L. infantum strains. • For the first known time L. infantum infection was recorded in ruminants (roe deer). • Over 90% of L. infantum infections in deer are associated with the "human" strain. • This result suggests that these animals are a novel reservoir for human visceral leishmaniasis. Leishmaniasis is a complex human disease caused by intracellular parasites of the genus Leishmania, predominantly transmitted by the bite of sand flies. In Italy, leishmaniasis is caused exclusively by Leishmania infantum , responsible for the human and canine visceral leishmaniases (HVL and CVL, respectively). Within the Emilia-Romagna region, two different foci are active in the municipalities of Pianoro and Valsamoggia (both in the province of Bologna). Recent molecular studies indicated that L. infantum strains circulating in dogs and humans are different, suggesting that there is an animal reservoir other than dogs for human visceral leishmaniasis in the Emilia-Romagna region. In this work, we analyzed specimens from wild animals collected during hunts or surveillance of regional parks near active foci of human visceral leishmaniasis for L. infantum infection in the province of Bologna. Out of 70 individuals analyzed, 17 (24%) were positive for L. infantum. The infection prevalence in hedgehogs (Erinaceus europaeus), roe deer (Capreolus capreolus), badgers (Meles meles), and bank voles (Myodes glareolus) was 80, 33, 25, and 11%, respectively. To distinguish the two strains of L. infantum we have developed a nested PCR protocol optimized for animal tissues. Our results demonstrated that most (over 90%) of L. infantum infections in roe deer were due to the strain circulating in humans in the Emilia-Romagna region. [ABSTRACT FROM AUTHOR]

Sand flies are hematophagous dipterans and are considered vectors of the parasites belonging to the genus Leishmania, which can infect mammals, including humans, and subsequently cause leishmaniasis. Rio Grande do Sul State (RS), southern Brazil, was considered free of this disease for the last few decades. However, several autochthonous cases of visceral leishmaniasis (VL) have been recorded in different municipalities in recent years. Since 2017, cases of canine visceral leishmaniasis (CVL) have been reported in the municipality of Santa Maria, located in the central region of RS. In 2021, two cases of human VL were confirmed in the city, resulting in one death. To capture the possible vectors involved in the disease transmission cycle, Centers for Disease Control (CDC) modified light traps were installed in the peridomestic residential areas. These points were situated in two city districts where cases of CVL had been reported from January 2021 to January 2022. In the 41 sample collections of sand flies, 9 specimens of Lutzomyia longipalpis were identified, confirming the municipality as an area of VL transmission, according to Brazilian sanitary standards. The spread of Leishmania spp. has been occurring efficiently, resulting in a negative impact on the One Health. This study reports the first detection of L. longipalpis in the central region of RS. [ABSTRACT FROM AUTHOR]

Leishmania infantum is the agent of visceral leishmaniasis in the Mediterranean basin. It is transmitted by sandflies of the subgenus Larroussius. Although Phlebotomus perniciosus is the most important vector in this area, an atypical Ph. perniciosus easily confused with Ph. longicuspis has been observed in North Africa. MALDI-TOF MS, an important tool for vector identification, has recently been applied for the identification of sandflies. Spectral databases presented in the literature, however, include only a limited number of Larroussius species. Our objective was to create an in-house database to identify Mediterranean sandflies and to evaluate the ability of MALDI-TOF MS to discriminate close species or atypical forms within the Larroussius subgenus. Field-caught specimens (n = 94) were identified morphologically as typical Ph. perniciosus (PN; n = 55), atypical Ph. perniciosus (PNA; n = 9), Ph. longicuspis (n = 9), Ph. ariasi (n = 9), Ph. mascittii (n = 3), Ph. neglectus (n = 5), Ph. perfiliewi (n = 1), Ph. similis (n = 9) and Ph. papatasi (n = 2). Identifications were confirmed by sequencing of the mtDNA CytB region and sixteen specimens were included in the in-house database. Blind assessment on 73 specimens (representing 1073 good quality spectra) showed a good agreement (98.5%) between MALDI-TOF MS and molecular identification. Discrepancies concerned confusions between Ph. perfiliewi and Ph. perniciosus. Hierarchical clustering did not allow classification of PN and PNA. The use of machine learning, however, allowed discernment between PN and PNA and between the lcus and lcx haplotypes of Ph. longicuspis (accuracy: 0.8938 with partial-least-square regression and random forest models). MALDI-TOF MS is a promising tool for the rapid and accurate identification of field-caught sandflies. The use of machine learning could allow to discriminate similar species. [ABSTRACT FROM AUTHOR]

Leishmaniasis on the Indian subcontinent is thought to have an anthroponotic transmission cycle. There is no direct evidence that a mammalian host other than humans can be infected with Leishmania donovani and transmit infection to the sand fly vector. The aim of the present study was to evaluate the impact of sand fly feeding on other domestic species and provide clinical evidence regarding possible non-human reservoirs through experimental sand fly feeding on cows, water buffalo goats and rodents. We performed xenodiagnosis using colonized Phlebotomus argentipes sand flies to feed on animals residing in villages with active Leishmania transmission based on current human cases. Xenodiagnoses on mammals within the endemic area were performed and blood-fed flies were analyzed for the presence of Leishmania via qPCR 48hrs after feeding. Blood samples were also collected from these mammals for qPCR and serology. Although we found evidence of Leishmania infection within some domestic mammals, they were not infectious to vector sand flies. Monitoring infection in sand flies and non-human blood meal sources in endemic villages leads to scientific proof of exposure and parasitemia in resident mammals. Lack of infectiousness of these domestic mammals to vector sand flies indicates that they likely play no role, or a very limited role in Leishmania donovani transmission to people in Bihar. Therefore, a surveillance system in the peri-/post-elimination phase of visceral leishmaniasis (VL) must monitor absence of transmission. Continued surveillance of domestic mammals in outbreak villages is necessary to ensure that a non-human reservoir is not established, including domestic mammals not present in this study, specifically dogs. Author summary: Leishmania donovani, the causative agent of visceral leishmaniasis (VL), has unique enzootic, zoonotic and anthroponotic life cycles dependent on geographic region. Most Leishmania spp. are zoonotic, transmitted between humans and non-human mammals. Leishmaniasis is endemic in over 98 countries and the estimated population at risk on the Indian sub-continent (ISC) is around 0.2–0.4 million people. Herein, we assess knowledge gaps in disease transmission that challenge dogma regarding the reservoir(s) of visceral leishmaniasis and control efforts on the Indian sub-continent. Better understanding of L. donovani infection in domestic animals and its transmission to sand flies will provide answers to fundamental questions in VL epidemiology and ecology. Blood samples were collected from livestock and rodents in endemic villages and colonized sand flies were fed on these animals and analyzed post feeding to see if they could pick up parasites via bloodmeal. We found that livestock and to a smaller extent rodents could be infected with or exposed to Leishmania infection. However, these mammals were not infectious to Phlebotomus argentipes, the predominant vector species incriminated as transmitting L. donovani in India. The inability of these domestic mammals to transmit L. donovani to sand flies in Bihar suggests that they play a limited role in the spread of infection. A key component in reducing and preventing the re-emergence of VL in the context of the elimination program is to understand the role of vector feeding on non-human animals in the transmission cycle. Better estimation of the proportion of the livestock and rodent population exposed to sand flies during different seasons and ensuring that there are not reservoir species that contribute to transmission will help implement appropriate control strategies for sustainable elimination. [ABSTRACT FROM AUTHOR]

Kala azar elimination programme in India is mainly depends on vector control measures and primary methods of application of insecticides is indoor residual spray (IRS). For vector control IRS should be carried out at regular intervals of six months. Presently, alphacypermethrin is used for IRS. This study was designed to study the impact of IRS with alphacypermethrin by analysing monthly population density following IRS-treatment. Twenty villages of Habibpur block, Malda district was selected for study which were grouped into study arm where IRS was done and control arm where IRS was not done. This study was conducted during April 2019 to June, 2020. In study villages, IRS was done during August, 2019 (1st cycle) and January 2020 (2nd cycle). Before and after IRS, entomological survey was carried out in cattle shades, human habitation and peri-domestic vegetations using mouth aspirators and light traps. Man hour density, per trap per night collection and % of reduction of sand fly abundance was calculated from survey result. In study arm after 14 days of 1st cycle of IRS, sand fly population reduced at rate of 96.5% which continues upto 3 months (85.34%). About 90% sand fly reduced after 14 days but after 3 months of 2nd cycle IRS density increased by 11.6%. Similar finding was recorded in the both cattle sheds and human habitations. Light trap collection from peri-domestic vegetation in both study and control villages were almost similar throughout the study period. Higher reduction rate of sand fly population following IRS showed that the used insecticide is effective against prevailing vector population of the study areas. The IRS has no effect on peri-domestic population. The presence of sand flies out side human habitation and cattle shades might play an important role in increasing population density following reduced residual activity of insecticide. [ABSTRACT FROM AUTHOR]

• The part of the insect used as a sample has an impact on the peptide/protein profile • Biological variables altered the profiles, generating identifiable diagnostic ions • When identifying sand flies with MALDI-TOF, biological factors must be considered • In the future, it will be possible to use MALDI-TOF to identify wild sand flies Sand flies are vectors of great public health importance, since they constitute a group of hematophagous insects responsible for etiological agents transmission of zoonotic diseases such a visceral leishmaniasis. In face of the expansion of these diseases, efficient control strategies are needed which depend on comprehending the sand fly eco-epidemiology. In this regard, MALDI-TOF mass spectrometry has been used for bacteria, fungi and yeast detection studies through peptide/protein profiles. However, little is known about interference of biological factors associated with vector ecology, such as blood meal preferences and even sand fly age on the peptide/protein profiles. Thus, the present study aimed to evaluate the differences in peptide/protein profiles of the sand fly Lutzomyia longipalpis , by means of MALDI-TOF, due to the sand fly's age, sex, blood meal source and Leishmania infantum infection. Sample preparation was made removing both head and last abdomen segments keeping the thorax, its appendices and the rest of the abdomen. Five specimens per pool were used to obtain peptide/protein extract of which 1 μL solution was deposited over 1 μL MALDI matrix dried. Characteristic spectra were analyzed using principal coordinate analysis as well as indicator species analysis to discriminate differences in sand flies's peptide/protein profile by sex, age, blood meal source and L. infantum infection. The results show that the evaluated variables produced distinct peptide/protein profiles, demonstrated by the identification of specific diagnostic ions. It was found that the interference of biological factors should be taken into account when using the MALDI-TOF analysis of sand fly species identification and eco-epidemiological applications in field studies. Based on our results, we believe that it is possible to identify infected specimens and the source of blood meal in a collection of wild sand flies, serving to measure infectivity and understand the dynamics of the vector's transmission chain. Our results may be useful for epidemiological studies that look at the ecology of sand flies and leishmaniasis, as well as for raising awareness of biological characteristics' impact on peptide/protein profiles in sand fly species identification. Experimental study design to evaluate the biological effects of age, blood feeding and Leishmania infantum infection in the sand fly species Lutzomyia longipalpis. [Display omitted] [ABSTRACT FROM AUTHOR]

Background: Most sand fly species are located in the Americas; some act as vectors of leishmaniasis and other human diseases. In Bolivia, about 25% of Neotropical species have been identified, and only a few have been implicated as vectors of cutaneous and visceral leishmaniasis. A new species of anthropophilic sand fly from the sub-Andean region of Alto Beni is described herein. Methods: A large systematic entomological survey was carried out in a subtropical humid forest located in the Marimonos mountain range, at around 900 m altitude, in the municipality of Palos Blancos, Sud Yungas Province, Department of La Paz, Bolivia. Sand flies were captured over a period of 26 months between January 1982 and February 1984, at the ground and canopy level, using both CDC light traps and protected human bait. A total of 24,730 sand flies were collected on the ground, distributed in 16 species, and 3259 in the canopy, with eight species. One of these species was labeled as Pintomia (Pifanomyia) nevesi, although certain morphological features allowed us to doubt that it was that taxon. To define the identity of this sand fly, a re-evaluation (this work) was recently carried out through morphological analyses and measurements of the available specimens mounted on Euparal, previously labeled as Pi. (Pif.) nevesi. Results: Based on the morphological traits and measurements, the re-evaluated specimens were definitively identified as a new sand fly species, Pintomyia (Pifanomyia) veintemillasi, closely related to Pi. (Pif.) nevesi and Pintomyia (Pifanomyia) maranonensis within the Evansi series. This new sand fly was the third most numerous anthropophilic species at the floor (6.2%) and the second most numerous anthropophilic at the canopy (35.1%). Conclusions: A new anthropophilic sand fly species is described as Pi. (Pif.) veintemillasi n. sp. This sand fly species was caught at about 900 m altitude in the Marimonos mountain range, a highly endemic area for cutaneous and mucosal leishmaniasis due to Leishmania (Viannia) braziliensis. Therefore, this species could be involved in the leishmaniasis transmission in the sub-Andean foothills of Alto Beni, Department of La Paz, Bolivia. [ABSTRACT FROM AUTHOR]

The use of repellents is considered an alternative against biting insects, including Lutzomyia longipalpis (Diptera: Psychodidae), the main vector of the protozoan Leishmania infantum, visceral leishmaniasis's (VL) etiologic agent in the Americas. This study aimed to evaluate the repellent efficacy of icaridin nanostructured solution applied on cotton knitting fabric against L. longipalpis. Arm-in-cage tests were performed in eight volunteers at different concentrations (5%, 10%, 25%, and 50%), using L. longipalpis (n = 30). The bioassay was performed in 1, 24, 48, 72, 96, 120, and 144 h after impregnation and one test after washing the fabrics with icaridin. The total repellency rate (%R) > 95% was used as a reference to define a minimum effective concentration (MEC). The results revealed that the insects' landing mean decreased significantly in different icaridin concentrations, compared with the control tests (p < 0.05) and the 25% and 50% concentrations compared to lower concentration (5%) (p < 0.05). The higher concentrations (25% and 50%) provided longer complete protection times (CPTs) with 120 and 144 h of protection, respectively and the %R of 100% for 72 and 96 h after impregnation, respectively. The 25% was the MEC (%R Total = 98.18%). Our results indicate, for the first time, that icaridin nanostructured solution applied on cotton knitting fabric proved to be an efficient repellent against L. longipalpis with the presence of repellent action even after washing. The concentration of 25% showed better efficiency and may become an efficient method for L. longipalpis biting control. [ABSTRACT FROM AUTHOR]

Background: Fleas (Insecta: Siphonaptera) are considered as highly specialized bloodsucking on mammals such as dogs. The existence of three factors, namely a vast distribution area, different hosts, and digestive system with a specific mechanism for digesting blood has led to species of fleas who nourish from mammals be introduced as the potential vectors of diseases. The aim of this study was to assess Leishmania infantum natural infection of dog fleas in northwest Iran in 2018. Methods: A total of 20 infested domestic dogs (Canis lupus familiaris) were randomly selected from 5 villages. Fleas were collected using brushing against dog hairs and fine forceps. Then, they were morphologically identified and preserved in ethanol for molecular assay. The kinetoplast DNA of the parasite was used for detection of Leishmania infantum using a semi-nested polymerase chain reaction (PCR) assay. Results: The human flea, Pulex irritans, and the cat flea, Ctenocephalides felis were identified on 40% and 35% of dogs, respectively. The results of PCR indicated that L. infantum was found in the Ctenocephalides canis (75%) and C. felis (66.7%) collected from infected dogs. No leishmanial infection was observed in P. irritans. Conclusion: It is concluded that fleas could be infected by Leishmania infantum, but maintenance of the parasite and their vectorial competence needs to be determined. [ABSTRACT FROM AUTHOR]

Leishmania spp. are sand fly‐borne protozoan parasites causing leishmaniasis in humans and animals. The aim of the study was to analyse the epidemiology of leishmaniasis in Turkey, Azerbaijan and Georgia from 2005 to 2020 and evaluate the associated risk for disease emergence in European countries. It is based on an analysis of WHO and OIE reported cases between 2005 and 2020, a review of scientific articles published in SCOPUS between 2009 and 2020 and a questionnaire survey to public health and veterinary authorities in these countries. Endemic Leishmania spp. include L. infantum in the three countries, L. major in Azerbaijan and Turkey and L. tropica and L. donovani in Turkey. Leishmaniasis is reported in humans, animals and sand flies and incidence is spatially and temporarily variable. In the southern Caucasus and particularly in Georgia, reported incidence of human visceral leishmaniasis by L. infantum remains high. However, whilst Georgia experienced a gradual decrease from >4.0 cases per 100,000 population in 2005–09 to 1.13 cases per 100,000 population in 2020, the period with highest incidence in Azerbaijan, which ranged between 0.40 and 0.61 cases per 100,000 population, was 2016–2019, and no cases have so far been reported for 2020. Visceral leishmaniasis in the Southern Caucasus affects mostly young children from deprived urban areas and its closely associated to canine leishmaniasis. Turkey reported cases of visceral leishmaniasis between 2005 and 2012 and in 2016 only, and incidence ranged between 0.02 and 0.05 per 100,000 population. In contrast, the reported annual incidence of cutaneous leishmaniasis in Turkey was much greater and peaked at 7.02 cases per 100,000 population in 2013, associated to imported cases from cutaneous leishmaniasis endemic Syria. Leishmaniasis by L. infantum in Azerbaijan and Georgia represents a regional public and animal health challenge that requires support to improve diagnosis, treatment and control. The unprecedented rise of cutaneous leishmaniasis and the spread of L. tropica and L. donovani in Turkey is an important risk factor for their emergence in Europe, especially in Mediterranean countries where competent vectors are widespread. [ABSTRACT FROM AUTHOR]

The development of Leishmania parasites within sand fly vectors occurs entirely in the insect gut lumen, in the presence of symbiotic and commensal bacteria. The impacts of host species and environment on the gut microbiome are currently poorly understood. We employed MiSeq sequencing of the V3-16S rRNA gene amplicons to characterize and compare the gut microbiota of field-collected populations of Phlebotomus kandelakii, P. perfiliewi, P. alexandri, and P. major, the primary or secondary vectors of zoonotic visceral leishmaniasis (ZVL) in three distinct regions of Iran where ZVL is endemic. In total, 160,550 quality-filtered reads of the V3 region yielded a total of 72 operational taxonomic units (OTUs), belonging to 23 phyla, 47 classes, 91 orders, 131 families, and 335 genera. More than 50% of the bacteria identified were Proteobacteria, followed by Firmicutes (22%), Deinococcus-Thermus (9%), Actinobacteria (6%), and Bacteroidetes (5%). The core microbiome was dominated by eight genera: Acinetobacter, Streptococcus, Enterococcus, Staphylococcus, Bacillus, Propionibacterium, Kocuria, and Corynebacterium. Wolbachia were found in P. alexandri and P. perfiliewi, while Asaia sp. was reported in P. perfiliewi. Substantial variations in the gut bacterial composition were found between geographically distinct populations of the same sand fly species, as well as between different species at the same location, suggesting that sand fly gut microbiota is shaped by both the host species and geographical location. Phlebotomus kandelakii and P. perfiliewi in the northwest, and P. alexandri in the south, the major ZVL vectors, harbor the highest bacterial diversity, suggesting a possible relationship between microbiome diversity and the capacity for parasite transmission. In addition, large numbers of gram-positive human or animal pathogens were found, suggesting that sand fly vectors of ZVL could pose a potential additional threat to livestock and humans in the region studied. The presence of Bacillus subtilis, Enterobacter cloacae, and Asaia sp suggests that these bacteria could be promising candidates for a paratransgenesis approach to the fight against Leishmaniasis. Author summary: Leishmania infantum, a parasitic protozoan causing fatal visceral leishmaniasis, is transmitted to humans by several sand fly vectors. In this study, the microbiota within the midguts of Phlebotomus kandelakii, P. perfiliewi, P. major and P. alexandri was analyzed by 16S ribosomal DNA (rDNA) Miseq sequencing, revealing highly diverse community composition and abundance, from three diverse ecological and geographical regions of Iran. It appears that the gut microbiota is highly dynamic and controlled by multiple factors, including sand fly host and environment. Proteobacteria were the principal bacterial phylum isolated. High numbers of gram-positive human or animal pathogens were also found, suggesting that sand fly vectors of ZVL could pose a potential threat to livestock and human in the region. Furthermore, there was a positive correlation between vector capacity and bacterial diversities, where the weakest ZVL vector had the lowest diversity, whereas other, more efficient, vectors had higher diversity. This study showed that Bacillus subtilis, Asaia sp. and Enterobacter cloacae are possible candidates for a paratransgenic approach to reduce Leishmania transmission. [ABSTRACT FROM AUTHOR]

Background: Visceral leishmaniasis (VL) is targeted for elimination as a public health problem in Nepal by 2023. For nearly three decades, the core vector control intervention in Nepal has been indoor residual spraying (IRS) with pyrethroids. Considering the long-term use of pyrethroids and the possible development of resistance of the vector Phlebotomus argentipes sand flies, we monitored the susceptibility status of their field populations to the insecticides of different classes, in villages with and without IRS activities in recent years. Methodology/Principal findings: Sand flies were collected from villages with and without IRS in five VL endemic districts from August 2019 to November 2020. The WHO susceptibility test procedure was adopted using filter papers impregnated at the discriminating concentrations of insecticides of the following classes: pyrethroids (alpha-cypermethrin 0.05%, deltamethrin 0.05%, and lambda-cyhalothrin 0.05%), carbamates (bendiocarb 0.1%) and organophosphates (malathion 5%). Pyrethroid resistance intensity bioassays with papers impregnated with 5× of the discriminating concentrations, piperonyl butoxide (PBO) synergist-pyrethroid bioassays, and DDT cross-resistance bioassays were also performed. In the IRS villages, the vector sand flies were resistant (mortality rate <90%) to alpha-cypermethrin and possibly resistant (mortality rate 90–97%) to deltamethrin and lambda-cyhalothrin, while susceptibility to these insecticides was variable in the non-IRS villages. The vector was fully susceptible to bendiocarb and malathion in all villages. A delayed knockdown time (KDT50) with pyrethroids was observed in all villages. The pyrethroid resistance intensity was low, and the susceptibility improved at 5× of the discriminating concentrations. Enhanced pyrethroid susceptibility after pre-exposure to PBO and the DDT-pyrethroid cross-resistance were evident. Conclusions/Significance: Our investigation showed that P. argentipes sand flies have emerged with pyrethroid resistance, suggesting the need to switch to alternative classes of insecticides such as organophosphates for IRS. We strongly recommend the regular and systematic monitoring of insecticide resistance in sand flies to optimize the efficiency of vector control interventions to sustain VL elimination efforts in Nepal. Author summary: Visceral leishmaniasis (VL), transmitted by P. argentipes sand flies, is endemic in South-East Asian countries such as Bangladesh, India, and Nepal, and is on the verge of elimination as a public health problem in Nepal by 2023. As part of the WHO Global Vector Control Response, entomological surveillance including insecticide resistance monitoring is one of the four main pillars of this strategy. In the early 1990s, the historical use of DDT for sand fly vector control was replaced with deltamethrin or alpha-cypermethrin, which have now been in use for almost three decades in Nepal. Suspecting that this long-term use of pyrethroids might have selected resistance in sand fly populations which would jeopardize control efforts, we conducted the first comprehensive survey to generate contemporary evidence of insecticide resistance in Nepal. For this, we performed WHO susceptibility tests in five VL endemic districts and found strong evidence of pyrethroid resistance in vector populations from the areas receiving IRS. Resistance mechanisms involved would probably be kdr mutations and monooxygenase. This study also endorses regular insecticide resistance monitoring to inform evidence-based decisions on insecticide use for vector control and to maintain the effectiveness of vector control measures as a core intervention in the fight against VL. [ABSTRACT FROM AUTHOR]

Leishmaniasis is a neglected tropical disease that continues to pose a serious public health problem. Albeit dogs have long been held as the major reservoirs of Leishmania infantum, the involvement of domestic cats in the zoonotic cycle of visceral leishmaniasis has gained prominence. Here, 240 cats were evaluated by clinical signs and haematological/biochemical changes compatible with leishmaniasis and were diagnosed by serological, molecular, and parasitological techniques. Thus, four cats naturally infected by L. infantum were submitted to xenodiagnosis. A total of 203 females of Lutzomyia longipalpis were subjected to feeding on four cats, with all females completing the blood meal. Parasitological and molecular assays were carried out to evaluate the presence of L. infantum in the sand flies' midgut. Promastigotes were observed in 10 females (6.5%) that fed on one cat, and L. infantum DNA was detected in 17 (8.4%) females that fed on two cats. Our results strengthen the evidence that naturally infected cats are capable of transmitting L. infantum to sand flies. [ABSTRACT FROM AUTHOR]

Background: In the State of Mato Grosso do Sul, Brazil, sand flies and cases of visceral (VL) and cutaneous (CL) leishmaniases have been reported in almost all municipalities. The aim of this study was to analyze the geographic distribution of VL and CL in relation the sand fly species found in the municipalities of Mato Grosso do Sul. Methods: We analyzed VL and CL cases from 2001 to 2018 using data from the Notifiable Diseases Information System (SINAN). Data collected since 2003 on the presence of sand fly vectors (proven or suspected) were provided by the State Health Secretariat. Results: A total of 3566 and 3030 cases of VL and CL, respectively, were reported from 2001 to 2018. The municipalities with the most reported cases of VL were Campo Grande (2495), Três Lagoas (442), Corumbá (140) and Aquidauana (136); and those for CL were Campo Grande (635) and Bodoquena (197). The following sand fly species with vector potential were found in 59 municipalities (74.7%): Lutzomyia longipalpis, Lutzomyia cruzi, Nyssomyia whitmani, Migonemyia migonei, Nyssomyia neivai, Pintomyia pessoai, Bichromomyia flaviscutellata and Pintomyia fischeri. Sand flies were present in six municipalities where no cases of VL were reported and in two municipalities where no cases of CL were reported. Conclusions: Our results indicate that the geographical distribution of VL and CL in Mato Grosso do Sul expanded during the study period, and highlight the presence of sand fly vectors in municipalities where these diseases are currently considered to be non-endemic. [ABSTRACT FROM AUTHOR]