Tsetse salivary glycoproteins are modified with paucimannosidic N-glycans, are recognised by C-type lectins and bind to trypanosomes.

African sleeping sickness is caused by Trypanosoma brucei, a parasite transmitted by the bite of a tsetse fly. Trypanosome infection induces a severe transcriptional downregulation of tsetse genes encoding for salivary proteins, which reduces its anti-hemostatic and anti-clotting properties. To better understand trypanosome transmission and the possible role of glycans in insect bloodfeeding, we characterized the N-glycome of tsetse saliva glycoproteins. Tsetse salivary N-glycans were enzymatically released, tagged with either 2-aminobenzamide (2-AB) or procainamide, and analyzed by HILIC-UHPLC-FLR coupled online with positive-ion ESI-LC-MS/MS. We found that the N-glycan profiles of T. brucei-infected and naïve tsetse salivary glycoproteins are almost identical, consisting mainly (>50%) of highly processed Man3GlcNAc2 in addition to several other paucimannose, high mannose, and few hybrid-type N-glycans. In overlay assays, these sugars were differentially recognized by the mannose receptor and DC-SIGN C-type lectins. We also show that salivary glycoproteins bind strongly to the surface of transmissible metacyclic trypanosomes. We suggest that although the repertoire of tsetse salivary N-glycans does not change during a trypanosome infection, the interactions with mannosylated glycoproteins may influence parasite transmission into the vertebrate host.

Insights into the salivary N-glycome of Lutzomyia longipalpis, vector of visceral leishmaniasis.

During Leishmania transmission sand flies inoculate parasites and saliva into the skin of vertebrates. Saliva has anti-haemostatic and anti-inflammatory activities that evolved to facilitate bloodfeeding, but also modulate the host's immune responses. Sand fly salivary proteins have been extensively studied, but the nature and biological roles of protein-linked glycans remain overlooked. Here, we characterised the profile of N-glycans from the salivary glycoproteins of Lutzomyia longipalpis, vector of visceral leishmaniasis in the Americas. In silico predictions suggest half of Lu. longipalpis salivary proteins may be N-glycosylated. SDS-PAGE coupled to LC-MS analysis of sand fly saliva, before and after enzymatic deglycosylation, revealed several candidate glycoproteins. To determine the diversity of N-glycan structures in sand fly saliva, enzymatically released sugars were fluorescently tagged and analysed by HPLC, combined with highly sensitive LC-MS/MS, MALDI-TOF-MS, and exoglycosidase treatments. We found that the N-glycan composition of Lu. longipalpis saliva mostly consists of oligomannose sugars, with Man5GlcNAc2 being the most abundant, and a few hybrid-type species. Interestingly, some glycans appear modified with a group of 144 Da, whose identity has yet to be confirmed. Our work presents the first detailed structural analysis of sand fly salivary glycans.

Cutaneous leishmaniasis and co-morbid major depressive disorder: A systematic review with burden estimates.

BACKGROUND: Major depressive disorder (MDD) associated with chronic neglected tropical diseases (NTDs) has been identified as a significant and overlooked contributor to overall disease burden. Cutaneous leishmaniasis (CL) is one of the most prevalent and stigmatising NTDs, with an incidence of around 1 million new cases of active CL infection annually. However, the characteristic residual scarring (inactive CL) following almost all cases of active CL has only recently been recognised as part of the CL disease spectrum due to its lasting psychosocial impact. METHODS AND FINDINGS: We performed a multi-language systematic review of the psychosocial impact of active and inactive CL. We estimated inactive CL (iCL) prevalence for the first time using reported WHO active CL (aCL) incidence data that were adjusted for life expectancy and underreporting. We then quantified the disability (YLD) burden of co-morbid MDD in CL using MDD disability weights at three severity levels. Overall, we identified 29 studies of CL psychological impact from 5 WHO regions, representing 11 of the 50 highest burden countries for CL. We conservatively calculated the disability burden of co-morbid MDD in CL to be 1.9 million YLDs, which equalled the overall (DALY) disease burden (assuming no excess mortality in depressed CL patients). Thus, upon inclusion of co-morbid MDD alone in both active and inactive CL, the DALY burden was seven times higher than the latest 2016 Global Burden of Disease study estimates, which notably omitted both psychological impact and inactive CL. CONCLUSIONS: Failure to include co-morbid MDD and the lasting sequelae of chronic NTDs, as exemplified by CL, leads to large underestimates of overall disease burden.

Cutaneous Leishmaniasis: The Truth about the 'Flesh-Eating Disease' in Syria.

Recent news headlines claimed that corpses thrown into Syrian streets are causing cutaneous leishmaniasis (CL) outbreaks. However, leishmaniasis is only transmitted by blood-feeding sandflies, not through human remains. High CL prevalence in Syria may instead be attributed to the absence of disease control programs due to the disruption of health services.

Severity of old world cutaneous leishmaniasis is influenced by previous exposure to sandfly bites in Saudi Arabia.

BACKGROUND: The sandfly Phlebotomus papatasi is the vector of Leishmania major, the main causative agent of Old World cutaneous leishmaniasis (CL) in Saudi Arabia. Sandflies inject saliva while feeding and the salivary protein PpSP32 was previously shown to be a biomarker for bite exposure. Here we used recombinant PpSP32 to evaluate human exposure to Ph. papatasi bites, and study the association between antibody response to saliva and CL in endemic areas in Saudi Arabia. METHODOLOGY/PRINCIPAL FINDINGS: In this observational study, anti-PpSP32 antibodies, as indicators of exposure to sandfly bites, were measured in sera from healthy individuals and patients from endemic regions in Saudi Arabia with active and cured CL. Ph. papatasi was identified as the primary CL vector in the study area. Anti-PpSP32 antibody levels were significantly higher in CL patients presenting active infections from all geographical regions compared to CL cured and healthy individuals. Furthermore, higher anti-PpSP32 antibody levels correlated with the prevalence and type of CL lesions (nodular vs. papular) observed in patients, especially non-local construction workers. CONCLUSIONS: Our findings suggest a possible correlation between the type of immunity generated by the exposure to sandfly bites and disease outcome.

Ampliación de la distribución de Lutzomyia longipalpis (Lutz & Neiva, 1912) (Diptera: Psychodidae) en el departamento de Caldas: potencial aumento del riesgo de leishmaniasis visceral / Expansion of the distribution of Lutzomyia longipalpis (Lutz & Neiva, 1912) (Diptera: Psychodidae) in the department of Caldas: Increased risk of visceral leishmaniasis

Introducción. Diversos estudios a nivel taxonómico y molecular sugieren que Lutzomyia longipalpis, principal vector de la leishmaniasis visceral en Latinoamérica, es un complejo de especies con amplia distribución a lo largo del continente y con adaptaciones a diferentes hábitats. Objetivo. Realizar vigilancia entomológica en el área de influencia de la hidroeléctrica Miel I. Materiales y métodos. Se utilizaron trampas adhesivas en una intersección de 400 km, aproximadamente, para la captura de insectos, los cuales fueron identificados con la clave taxonómica de Young y Duncan. Resultados. En este estudio se reporta la presencia de Lu. longipalpis en los municipios de Norcasia a una altitud de 392 msnm y en Marquetalia a 1.387 msnm; es el primer reporte a esta altura para esta especie en Colombia. Conclusiones. Se sugiere que la ampliación de la distribución geográfica y altitudinal de Lu. longipalpisen el país podría estar relacionada con la existencia del complejo de especies Lu. longipalpis, o a factores ambientales como el aumento de temperatura debido al cambio climático global que viene sucediendo en las últimas décadas. Esta última hipótesis sugeriría que se están creando nuevos hábitats propicios para el establecimiento de Lu. longipalpis, lo cual genera un nuevo riesgo epidemiológico de posibles nuevos focos de leishmaniasis visceral en el país.

Introduction: Diverse taxonomical and molecular studies suggest that Lutzomyia longipalpis, main vector of visceral leishmaniasis in Latin America, is a complex of species widely distributed throughout the continent and adapted to different habitats. Objective: To carry out entomological surveillance in the area of influence of the Miel I Hydroelectric Plant. Materials and methods: Adhesive traps were used in a transect covering an area of approximately 400 km for the capture of insects, which were then identified using the Young and Duncan taxonomical key. Results: This study reports the presence of Lutzomyia longipalpis in the municipalities of Norcasia at an altitude of 392 masl, and in Marquetalia at 1,387 masl. Conclusions: We suggest that the increase in the geographical and altitudinal distribution could be related to the existence of the Lu. longipalpis species complex, or to environmental factors, such as the rise in temperature due to global climate changes, which create the establishment of Lu. longipalpis, generating a new epidemiological risk for new visceral leishmaniasis foci in the country.

[Expansion of the distribution of Lutzomyia longipalpis (Lutz & Neiva, 1912) (Diptera: Psychodidae) in the department of Caldas: Increased risk of visceral leishmaniasis]. / Ampliación de la distribución de Lutzomyia longipalpis (Lutz & Neiva, 1912) (Diptera: Psychodidae) en el departamento de Caldas: potencial aumento del riesgo de leishmaniasis visceral.

INTRODUCTION: Diverse taxonomical and molecular studies suggest that Lutzomyia longipalpis , main vector of visceral leishmaniasis in Latin America, is a complex of species widely distributed throughout the continent and adapted to different habitats. OBJECTIVE: To carry out entomological surveillance in the area of influence of the Miel I Hydroelectric Plant. MATERIALS AND METHODS: Adhesive traps were used in a transect covering an area of approximately 400 km for the capture of insects, which were then identified using the Young and Duncan taxonomical key. RESULTS: This study reports the presence of Lutzomyia longipalpis in the municipalities of Norcasia at an altitude of 392 masl, and in Marquetalia at 1,387 masl. CONCLUSIONS: We suggest that the increase in the geographical and altitudinal distribution could be related to the existence of the Lu. longipalpis species complex, or to environmental factors, such as the rise in temperature due to global climate changes, which create the establishment of Lu. longipalpis , generating a new epidemiological risk for new visceral leishmaniasis foci in the country.

Cutaneous leishmaniasis in the dorsal skin of hamsters: a useful model for the screening of antileishmanial drugs.

Traditionally, hamsters are experimentally inoculated in the snout or the footpad. However in these sites an ulcer not always occurs, measurement of lesion size is a hard procedure and animals show difficulty to eat, breathe and move because of the lesion. In order to optimize the hamster model for cutaneous leishmaniasis, young adult male and female golden hamsters (Mesocricetus auratus) were injected intradermally at the dorsal skin with 1 to 1.5 x l0(7) promastigotes of Leishmania species and progression of subsequent lesions were evaluated for up to 16 weeks post infection. The golden hamster was selected because it is considered the adequate bio-model to evaluate drugs against Leishmania as they are susceptible to infection by different species. Cutaneous infection of hamsters results in chronic but controlled lesions, and a clinical evolution with signs similar to those observed in humans. Therefore, the establishment of the extent of infection by measuring the size of the lesion according to the area of indurations and ulcers is feasible. This approach has proven its versatility and easy management during inoculation, follow up and characterization of typical lesions (ulcers), application of treatments through different ways and obtaining of clinical samples after different treatments. By using this method the quality of animal life regarding locomotion, search for food and water, play and social activities is also preserved.