CLIMOS Open Source Publications
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Authors: Nazli Ayhan, Cecile Baronti, Laurence Thrion, Gioia Bongiorno, Carla Maia, Remi N. Charrel & CLIMOS WP2 Phlebovirus Working Group
Abstract
Background
Sand fly-borne phleboviruses (SbPV) are globally distributed and pose potential public health risks. Despite increased detection in recent decades, detailed knowledge of their ecology, characteristics and clinical relevance remains limited. Many cases of SbPV infection likely go unreported or misdiagnosed due to limited awareness and the lack of standardized screening. The External Quality Assessment (EQA) reported herein was organized within the framework of the European Union CLIMOS (EU Climate Monitoring and Decision Support Framework for Sand Fly-borne Diseases Detection and Mitigation) project. The aim of this EQA was to standardize the detection of phleboviruses in order to provide comparable data to feed mathematical models for the surveillance of the impact of climate changes and environmental parameters on the kinetics and diversity of sand fly species and on sand fly-borne microorganisms.
Methods
Nine laboratories from seven countries participated in the EQA. Each laboratory was provided with eight vials, each containing an anonymous sample; two vials of lyophilized primers and probes to be used for the detection of Toscana virus (TOSV) and several Sandfly fever Sicilian virus (SFSV) species with a reverse-transcriptase PCR (RT-PCR) assay; and one vial of lyophilized primers for the detection of generic phleboviruses with a RT-PCR assay along with the standard operating procedure. The laboratories were instructed to submit their results together with details on the techniques employed.
Results
All nine laboratories successfully detected the two TOSV- and the one SFSV-positive samples. Only one laboratory, using a generic phlebovirus assay, detected all of the targeted phleboviruses.
Conclusions
All participating laboratories successfully identified the two TOSV and one SFSV using the proposed RT-qPCR assays, albeit with some variations in cycle threshold values across laboratories. The detection rate of SbPV was lower with the generic Phlebovirus assay than with the specific real-time RT-qPCR assays. This EQA aimed to assess the SbPV detection capabilities of molecular tools and strengthen their use, thereby supporting the involvement of laboratories in virus discovery and surveillance beyond their core expertise.
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Authors: Jorian Prudhomme, Aymeric Delabarre, Bulent Alten, Umut Berberoglu, Eduardo Berriatua, Gioia Bongiorno, José Manuel Cristovao, Maya Davidovich-Cohen, Trentina Di Muccio, Ozge Erisoz Kasap, Eleonora Fiorentino, Oscar D Kirstein Edwin Kniha, Carla Maia, Mesut Mungan, Clara Muñoz-Hernández, Muhammed Nalçaci, Gizem Oguz Kaskan, Yusuf Ozbel, Seray Ozensoy Toz, Ricardo Parreira, Katharina Platzgummer, Ceylan Polat, José Risueño, Liora Studentsky, Gamze Varol, Julia Walochnik, Kardelen Yetişmiş, Florence Robert-Gangneux
Abstract
Background: Leishmaniasis, caused by Leishmania protozoan parasites transmitted by Phlebotomine sand flies, is a significant public health concern in the Mediterranean basin. Effective monitoring of Leishmania-infected sand flies requires standardized tools for comparing their distribution and infection prevalence. Consistent quantitative real-time PCR (qPCR) parameters and efficient DNA extraction protocols are crucial for reliable results over time and across regions. However, the absence of standardized technical recommendations for Leishmania DNA detection hinders effective surveillance. This study aimed to compare different DNA extraction protocols and conduct a qPCR-based External Quality Assessment (EQA) through a multicenter study involving nine reference laboratories, with a focus on optimizing Leishmania DNA detection in sand fly.
Methodology/principal findings: EQA samples consisted of Leishmania infantum and L. major species, at concentrations ranging from 101 to 104 parasites/mL. All but one center detected all concentrations, demonstrating strong diagnostic proficiency. The ability to detect low concentrations highlighted the robustness of the qPCR assay used, though variations in Cq values indicated differences in sensitivity related to technical capabilities or DNA extraction kit performance. A comparative analysis of seven DNA extraction methods identified the EZ1 DSP Virus Kit and QIAamp DNA mini-kit as the most efficient, supporting their use in standardized protocols. The study also assessed the effects of lyophilization and shipment conditions, showing no significant compromise in Leishmania detection despite slight variations in Cq values. Experimentally infected sand flies were included to simulate field conditions, and all centers successfully detected positive samples with varying Cq values, probably reflecting differences in infection load.
Conclusion and significance: This study emphasizes the importance of standardized DNA extraction protocols and continuous quality assurance for accurate Leishmania DNA detection. The results highlight the superior performance of certain extraction kits and the need for ongoing technical training, essential for reliable leishmaniasis surveillance, particularly in field settings with low infection densities.
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Authors:
Jorian Prudhomme , Jérôme Depaquit, and Florence Robert-Gangneux
Abstract
Global changes in climate are contributing to modified Phlebotomine sand fly presence and activity, and the distribution of the pathogens they transmit (e.g., Leishmania and Phlebovirus), and are leading to their possible extension toward northern France. To predict the evolution of these pathogens and control their spread, it is essential to identify and characterize the presence and abundance of potential vectors. However, there are no recent publications describing sand fly species distribution in France. Consequently, we carried out a systematic review to provide distri- bution and abundance maps over time, along with a simplified dichotomous key for species in France. The review adhered to PRISMA guidelines, resulting in 172 relevant capture reports from 168 studies out of the 2646 documents retrieved, of which 552 were read and 228 analyzed. Seven species were recorded and categorized into three groups based on their abundance: low abundance species, abundant but little-studied species, and abundant vector species. Sand flies are certainly present throughout France but there is a greater diversity of species in the Mediterranean region. Phlebotomus perniciosus and Ph. ariasi are the most abundant and widely distributed species, playing a role as vectors of Leishmania. Sergentomyia minuta, though very abundant, remains under-studied, highlighting the need for further research. Phlebotomus papatasi, Ph. perfiliewi, Ph. sergenti, and Ph. mascittii are present in low numbers and are less documented, limiting understanding of their potential role as vectors. This work provides the necessary basis for comparison of field data generated in the future.
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Authors:
Bruno M. Carvalho, Carla Maia, Orin Courtenay, Alba Llabrés-Brustenga, Martín Lotto Batista, Giovenale Moirano, Kim R. van Daalen, Jan C. Semenza, Rachel Lowe.
Abstract
Background
Methods
Findings
Interpretation
Funding
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Authors:
Alejandro Polina, Yasmina Martínez-Barciela, José Manuel Pereira, Fernando Cobo, Elvira Íñiguez, Javier Lucientes, Rosa Estrada, Josefina Garrido
Abstract
Introduction
Culicoides Latreille biting midges are vectors of high concern as they can transmit serious veterinary diseases such as bluetongue virus or epizootic haemorrhagic disease virus, among others. Little is known about these vectors in Galicia, so a comprehensive literature review and an intensive monitoring were carried out in the region.
Material and methods
The Autonomous Community of Galicia was sampled through three different vector surveillance projects between 2004 and 2023. A total of 239 sampling points were deployed alongside the Galician territory. In addition, a literature review of Culicoides in Galicia related content was made by consulting several digital repositories.
Results
A total of 33 species of Culicoides belonging to 8 subgenera were identified. Among them, 15 are considered or suspected to be potential vectors of several pathogens of medical and/or veterinary interest. In addition, 20 of them are reported for the first time in the region. Updated distribution maps of the Culicoides biting midges of Galicia were provided, including several notes regarding their ecology and relevance for both public health and animal welfare.
Conclusions
The present work is one of the most complete works made at regional level in Spain to date. As Galicia’s economy relies heavily on livestock farming, this work will provide a solid baseline in order to develop new research lines in the future regarding prevention to vector-borne diseases.
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Authors: Tarcisio de Freitas Milagres, Carla Maia
Abstract
Phlebotomus perniciosus, recognized as the most important phlebotomine sand fly vector in countries of the Western Mediterranean region, is responsible for transmitting the protozoan Leishmania infantum, the causative agent of zoonotic leishmaniasis. This species also serves as a vector for various phleboviruses, with the Toscana virus being the most clinically relevant, associated as one of the main causes of meningitis and encephalitis in this region. Detected in 22 countries, P. perniciosus has a broad distribution in countries located in the Western part of the Mediterranean basin. However, due to global changes, it is expected that it will expand its endemicity to currently non-endemic regions. In endemic areas, this species is usually considered the most abundant and predominant vector, except in colder and more humid regions. It can be found from May to November, with density trends that may vary with each endemic scenario, making its presence temporally heterogeneous.
Published in Trends in Parasitology
DOI: 10.1016/j.pt.2024.04.007
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Authors:
Eduardo Berriatua, Pedro Pérez‐Cutillas, Aurora González Vidal, and Olivier J. T. Briët
Abstract
Background
Leishmania infantum is endemic in Europe (and elsewhere) while L. donovani s.s., L. tropica and L. major are not but are present in neighboring countries in North Africa, the Middle East, (the Asian part of ) Turkey and the Southern Caucasus. Lists of sand fly vector species in the scientific literature vary with the criteria for vector incrimination, and criteria vary because, for some, evidence is difficult to generate. With minimal criteria, about 20 sand fly species are proven or suspected vectors of L. infantum in Europe and neighboring countries, while for L. trop- ica and L. major, there are seven and four proven or suspected vector species, respectively, in this area. For L. donovani s.s., present in Cyprus, the Middle East and (the Asian part of ) Turkey, no local vectors have been incriminated so far. The aim was to assess the degree of spatial agreement between Leishmania spp. and various vectors species and their relative contribution to the explained variation.
Methods
We used multivariate regression modeling to analyze the spatial relationship between autochthonous Leishmania spp. and clinical forms in humans and animals and 14 Phlebotomus spp. in Europe and neighboring countries.
Results
There was only fair agreement between parasite and vector distributions. The most parsimonious models describing the distribution of Leishmania spp. and clinical forms included three to six sand fly species and explained between 12% (L. infantum) and 37% (L. donovani) of the observed variation. Selected models included confirmed and suspected vector species as well as unexpected species.
Conclusions
The relatively low agreement between Leishmania and vector distributions highlights the need to improve leishmaniasis reporting and vector surveillance in areas where no information is available, both for a better understanding of the epidemiology of infection in endemic areas and to monitor possible spread of infection into non‐endemic areas. While some of the unexpected sand fly‐Leishmania spp. statistical associations might be spu‐ rious, for others, the existence of sporadic or recent reports of infections warrants further vector competence studies that consider strain variation.
DOI: 10.1186/s13071-024-06484-2
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Authors:
Tarcísio de Freitas Milagres, Carla Maia
Young Reviewers:
Greshma, Eugene, and Navinder
Abstract
DOI: 10.3389/frym.2024.1422953
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Authors:
Anna Hošková, Barbora Vojtková, Markéta Stejskalová, Nikola Polanská, Magdalena Jančářová, Lidiane Medeiros da Costa, Mauricio Roberto Viana Sant Anna, Petr Volf, Jovana Sádlová
Abstract
Background: We evaluated various membranes for blood-feeding in nine sand fly species from different genera and subgenera. Most of these species are vectors of human-pathogenic Leishmania, whereas Sergentomyia minuta is a herpetophilic sand fly species and a proven vector of Leishmania (Sauroleishmania) tarentolae.
Methods: Female sand flies were offered blood through a range of membranes (chicken, reptilian, and frog skin; synthetic collagen; pig intestine; and duck foot webbing). Two feeding systems (glass feeder, Hemotek) and different blood sources (human, ovine, avian, and reptilian) were used. Feeding trials were conducted under varying thermal and light conditions to determine the optimal parameters.
Results: Among the 4950 female S. minuta tested, only a negligible fraction took a blood meal: 2% of the females fed on avian blood, and 0.2% of the females fed on human blood. In eight other species, the chicken membrane was generally more effective than synthetic membranes or pig intestines. For example, Phlebotomus duboscqi refused synthetic membranes, while Lutzomyia longipalpis and P. perniciosus avoided both synthetic membranes and pig intestines. The most effective membrane was duck foot webbing, with four species feeding more readily through it than through the chicken membrane. Additionally, applying coagulated blood plasma to the outer surface of chicken or synthetic membranes significantly increased feeding rates.
Conclusions: Female S. minuta did not reliably feed on blood through the tested membranes, preventing laboratory infection experiments from confirming their vector competence for human-pathogenic Leishmania. However, for future experimental infections of other sand fly species, duck foot webbing has emerged as an effective membrane, and the application of blood plasma to the exterior of membranes may increase the feeding rates.
DOI: 10.1186/s13071-025-06729-8
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Authors:
Mohammad Akhoundi, Michele Maroli, Luigi Gradoni, Arezki Izri, Jean-Pierre Dedet, Jan Votýpka, Souad Guernaoui, Razika Benikhlef, Naouel Eddaikra, Paul Bates, Denis Sereno & Jérôme Depaquit
Abstract
Given the significance of leishmaniasis as a neglected parasitic disease—ranking second in mortality and fourth in morbidity among vector-borne diseases—and the prominence of the Mediterranean and Middle East regions as key areas for leishmaniasis incidence, the study and precise morphological identification of sand flies, the proven vectors of the disease, is crucial. Unfortunately, despite this importance, there are few reliable references or identification keys for the morphological identification of sand flies in the Middle East and Mediterranean regions. Some are outdated and no longer valid or remain local (restricted to a country). To date, no comprehensive study has been conducted on the sand fly fauna and their morphological characterization across these regions. In response to this gap, we present a comprehensive pictorial identification key for male and female Phlebotomus species of Middle East and Mediterranean areas. The key includes 720 selected photos and illustrations demonstrating discriminative morphological features out of 2,000 collected. Furthermore, a collection including descriptive morphological criteria of sand flies, first description of Phlebotomus species, a comprehensive checklist of Phlebotomus species accompanied by their distribution map across Middle Eastern and Mediterranean countries, as well as extensive information on their morphometry, ecology, medical relevance, synonymy, atypical forms and morphology of female Adlerius species are given. Finally, we provide an online pictorial dichotomous key to facilitate field application.
DOI: 10.1038/s41598-024-77815-7
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Authors:
Eunice Aparecida Bianchi Galati, Andrey José de Andrade, Farzana Perveen, Mathieu Loyer, Khamsing Vongphayloth, Fano José Randrianambinintsoa, Jorian Prudhomme, Nil Rahola, Mohammad Akhoundi, Paloma Helena Fernandes Shimabukuro & Jérôme Depaquit
Abstract
Abstract
Background
Checklists of zoological groups are useful to document species names in a specific area or even worldwide. They serve for various purposes, including ecological studies, conservation reports, policy and decision-making, and species identification. Phlebotomine sand flies (Diptera, Psychodidae) are the vectors of pathogens such as Leishmania, Bartonella and some arboviruses (Toscana, Naples, Sicily), and checklists for sand flies have primarily been published mainly for a state, department, or country. A checklist for American sand flies was published in 2017, but, until then, no effort has been made to compile a comprehensive list of species worldwide. The present study aims to fill this gap of knowledge.
Methods
The present checklist is provided based on a literature overview and biological collections records and includes unpublished data from the authors. The species are presented according to the classification, then alphabetically by Eastern and Western Hemispheres. Distribution by country and type locality of each species are provided. Discussions on the taxonomic status or occurrence of each species are provided when needed.
Results
A total of 23 genera in the Western Hemisphere, formerly the New World (Bichromomyia, Brumptomyia, Deanemyia, Dampfomyia, Edentomyia, Evandromyia, Expapillata, Hertigia, Lutzomyia, Martinsmyia, Micropygomyia, Migonemyia, Nyssomyia, Oligodontomyia, Pintomyia, Pressatia, Psathyromyia, Psychodopygus, Sciopemyia, Trichophoromyia, Trichopygomyia, Viannamyia, and Warileya) and 17 genera in the Eastern Hemisphere, formerly the Old World (Australophlebotomus, Chinius, Demeillonius, Grassomyia, Idiophlebotomus, Libanophlebotomus, Libanophlebotomites, Mesophlebotomites, Paleomyia, Parvidens, Phlebotoiella, Phlebotomiella, Phlebotomites, Phlebotomus, Sergentomyia, Spelaeomyia, Spelaeophlebotomus), including the fossil species, are listed herein. The updated list, excluding nomina dubia/species inquirenda includes 1063 sand fly species, with 549 and 514 species for New World and Old World, respectively. Only New Zealand and the Pacific Islands (excluding New Caledonia) do not record the presence of sand flies. The dataset for this study is publicly available in the SiBBr and GBIF.
Conclusions
This is the first detailed list of valid names of phlebotomine sand flies worldwide, including records from each country where they have been documented.
DOI: 10.1186/s13071-025-06748-5
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Authors:
Jovana Sadlova , Anna Hoskova, Barbora Vojtkova, Tomas Becvar, Petr Volf
Abstract
Leishmaniases are serious human and animal diseases caused by protozoa of the genus Leishmania (Kinetoplastida: Trypanosomatidae), transmitted by blood-feeding insects – phlebotomine sand flies (Diptera: Psychodidae). They are mainly distributed in tropical and subtropical regions, but recently the ranges of the vectors have spread more into temperate regions. In Europe, species of Leishmania that were previously absent have emerged, mainly due to the mass migration of the refugees from endemic areas. It is therefore important to know whether local sand flies can transmit these “new” Leishmania species. In our study, we tested two widely distributed sand fly species, P. perniciosus and P. tobbi. In both of them, L. major and L. donovani completed their development, thus P. perniciosus and P. tobbi are potential vectors of these parasites. The third Leishmania species tested, L. martiniquensis, which has been repeatedly recorded in Central Europe, did not survive in P. perniciosus and P. tobbi; its vectors have yet to be confirmed.
DOI: 10.1371/journal.pntd.0012597
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Authors:
Edwin Kniha, Stephan Koblmüller, Katharina Platzgummer, Oscar Kirstein, Debora Diaz, Vít Dvořák, Ozge Erisoz Kasap, Betim Xhekaj, Kurtesh Sherifi, Julia Walochnik & Attila J. Trájer
Abstract
Phlebotomine sand flies (Diptera: Psychodidae: Phlebotominae) are the principal vectors of Leishmania spp. (Kinetoplastida: Trypanosomatidae) worldwide. The subgenus Adlerius is taxonomically challenging and currently comprises about 20 species with a wide geographic distribution from eastern Asia to southeastern Europe. Some species are confirmed or suspected vectors of Leishmania donovani/infantum, L. major, and L. tropica, and are thus of high medical and veterinary relevance. A single record of Phlebotomus (Adlerius) simici in Austria from 2018 marks its sporadic northernmost and westernmost occurrence, with the origin of its appearance remaining unclear. To better understand Adlerius diversification and particularly post-glacial spread of Ph. simici to northern parts of Europe, we combined phylogenetic analyses with climatic suitability modelling. Divergence time estimates well supported the currently observed geographic distribution of the studied species and revealed several taxonomic challenges in the subgenus. We clearly delineated three distinct genetic and geographic Ph. simici lineages and phylogeographically assessed diversification that were well supported by climatic models. This study provides a comprehensive phylogenetic analysis of the subgenus Adlerius, enhancing our understanding of the diversification in relation to changing climate of this understudied group, and we present new insights into the post-glacial spread of Ph. simici, a suspected vector of L. infantum.
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Authors:
Rafael Rocha, Elif Kurum, Nazli Ayhan, Rémi Charrel & Carla Maia
Abstract
Background
Toscana virus (TOSV; Phlebovirus toscanaense), a phlebovirus transmitted by sand flies, is a growing public health concern in the Mediterranean region, with infections often being asymptomatic but potentially leading to neuroinvasive disease. Despite its presence in neighboring countries, data on TOSV seroprevalence in Portugal are limited. This study aimed to estimate the national seroprevalence of TOSV among blood donors in mainland Portugal and explore associations with sociodemographic factors and Leishmania infection.
Methods
A cross-sectional study was conducted using serum samples from 3593 blood donors across mainland Portugal, collected between February and June 2022. Anti-TOSV antibodies were detected via microneutralization assay, and anti-Leishmania antibodies had previously been tested using ELISA. Sociodemographic data were obtained from self-administered questionnaires. Seroprevalence was estimated by region, and multivariate logistic regression was used to identify factors associated with TOSV infection.
Results
Overall, the estimated national true seroprevalence of TOSV was 2.6% (95% CI 2.1–3.1%). Regional seroprevalence varied significantly, with the highest values (up to 14.8%) in Alto Alentejo, Baixo Alentejo, Douro, Alto Tâmega e Barroso and Oeste regions. Multivariate analysis showed that age ≥ 50 years (aOR 1.70, 95% CI 1.04–2.77), residing in the Alentejo region (aOR 3.05, 95% CI 1.85–5.02) and positive/borderline Leishmania serology (aOR 2.31, 95% CI 1.29–4.15) were significantly associated with TOSV infection.
Conclusions
This study highlights new areas of TOSV circulation in Portugal, particularly in regions with higher Leishmania seroprevalence and visceral leishmaniasis incidence, suggesting co-circulation of these pathogens. Although a lower seroprevalence was obtained compared to neighboring countries, TOSV should still be considered in the differential diagnosis of viral meningitis and encephalitis in Portugal, especially in potentially high-risk regions. Further research is needed to better understand the ecological drivers of TOSV distribution in Portugal.
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Authors: Rafael Rocha, Cláudia Conceição, Luzia Gonçalve, LeishPT group and Carla Maia
Abstract
Background: Leishmania infantum is endemic in the Mediterranean region, presenting mostly as visceral leishmaniasis (VL). In Portugal, reporting of VL cases to public health authorities is mandatory, but significant underreporting is likely. This study aimed to describe the epidemiological and clinical aspects of the VL cases diagnosed in hospitals of the Portuguese National Health Service (NHS), between 2010 and 2020.
Methods: Collaboration was requested to every hospital of the Portuguese NHS in Mainland Portugal. Cases were screened through a search of diagnostic discharge codes or, if not available, by a search of positive laboratory results for Leishmania infection. Sociodemographic and clinical data was retrieved from medical records. Simultaneously, the National Health authority was contacted to request access to data of notified cases of VL between 2010 and 2020. Descriptive, hypothesis testing and multiple binary logistic regression models were performed.
Results: A total of 221 VL cases were identified. A significant increase in estimated national incidence was seen in the years after 2016 (P = 0.030). VL was predominantly diagnosed in people living with HIV (PLWH) and in children (representing around 60% of the new cases), but the outcome was generally poorer in non-HIV patients with associated immunosuppression, with significantly lower rates of clinical improvement at 7 (P = 0.003) and 30 days (P = 0.008) after treatment. Atypical presentations, with gastrointestinal and/or respiratory involvement, were seen in 8.5% of VL cases. Hemophagocytic lymphohistiocytosis was diagnosed in 40.0% of children under 5 years of age. Only 49.7% of incident VL cases were reported. Simultaneous involvement of the skin was confirmed in 5.9% of patients.
Conclusions: VL presents a continuing threat in Portugal, especially to PLWH and children, and an increasing threat to other immunosuppressed groups. Recent increases in incidence should be closely monitored to allow prompt interventions. Programs to control the disease should focus on providing tools for earlier diagnosis and on reducing underreporting and promoting an integrated surveillance of human and animal disease. These data should be combined with asymptomatic infection and vector information, following a One Health approach.
Published in Trends in Parasitology
DOI: 10.1186/s40249-024-01204-5
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Authors:
Magdalena Jancarova, Nikola Polanska, Adrien Thiesson, Frédérick Arnaud, Marketa Stejskalova, Marketa Rehbergerova, Alain Kohl, Barbara Viginier, Petr Volf, Maxime Ratinier
Abstract
Sand flies are blood-sucking insects that transmit various human pathogens, including parasitic protists of the genus Leishmania and several viruses, such as the phlebovirus Toscana (TOSV). TOSV infections typically result in flu-like symptoms, but in some patients, the infection can affect the central nervous system (CNS) or even be fatal. Due to climatic and environmental changes associated with human activity, it is likely that sand flies and the pathogens they transmit will spread beyond current boundaries. However, the specific sand fly species that transmit TOSV are not well understood. Therefore, it is essential to identify vectors that are involved in the circulation and transmission of the virus. In our study, we found that the range of potential TOSV vectors is broader than previously thought and that individual virus strains may influence vector competence. Specifically, we demonstrated that Phlebotomus tobbi and P. sergenti, two sand flies common in Mediterranean area and Middle East, are susceptible to TOSV and likely transmit this virus in nature.
DOI: 10.1371/journal.pntd.0013031
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Authors: Rafael Rocha, Cláudia Conceição, Luzia Gonçalves, LeishPT group: Ana Cláudia Carvalho, André Maia, André Martins, António Carujo, António Maio, Catarina Forra, Catarina Melita, Daniela Couto, Diana Fernandes, Dulce Pereira, Ema Leal, Helena Sarmento, Inês Sousa, Jean-Pierre Gonçalves, Joana Marinho, Joana Vasconcelos, João Cunha, João Rodrigues, José Miguel Silva, Lídia Caley, Luís Malheiro, Luís Santos, Margarida Garcia, Margarida Prata, Maria Cunha, Maria Lima, Maria Margarida Andrade, Marta Marques, Miguel Alpalhão, Mónica Silva, Rita Ferraz, Rui Soares, Salomão Fernandes, Samuel Llobet, Sofia Cruz, Teresa Guimarães, Tiago Branco, Tomás Robalo-Nunes, Vasco Almeida, Carla Maia
Abstract
Leishmania infantum, a zoonotic vector-born parasite, is endemic in the Mediterranean region, presenting mostly as visceral (VL), but also as cutaneous (CL) and mucosal leishmaniasis (ML). This study aimed to describe the epidemiological and clinical aspects of the CL and ML cases diagnosed in mainland Portugal between 2010 and 2020. Collaboration was requested from every hospital of the Portuguese National Health System. Cases were screened through a search of diagnostic discharge codes or positive laboratory results for Leishmania infection. Simultaneously, a comprehensive literature search was performed. Descriptive statistics and hypothesis testing were performed using IBM® SPSS® Statistics. A total of 43 CL and 7 ML cases were identified, with a predominance of autochthonous cases (86%). In CL, immunosuppressed individuals constituted a significant proportion of patients (48%), and in this group, disseminated CL (22%) and simultaneous VL (54%) were common. In autochthonous cases, lesions, mostly papules/nodules (62%), were frequently observed on the head (48%). The approach to treatment was very heterogeneous. ML cases were all autochthonous, were diagnosed primarily in older immunosuppressed individuals, and were generally treated with liposomal amphotericin B. The findings suggest a need for enhanced surveillance and reporting, clinical awareness, and diagnostic capacity of these forms of leishmaniasis to mitigate underdiagnosis and improve patient outcomes. A holistic One Health approach is advocated to address the multifaceted challenges posed by leishmaniases in Portugal and beyond.
Published in Microorganisms
DOI: 10.3390/microorganisms12040819
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Authors: Kim R van Daalen, PhD Prof Cathryn Tonne, ScD Prof Jan C Semenza, PhD Prof Joacim Rocklöv, PhD Prof Anil Markandya, PhD Niheer Dasandi, PhD Prof Slava Jankin, PhD Hicham Achebak, PhD Joan Ballester, PhD Hannah Bechara, PhD Thessa M Beck, MSc Max W Callaghan, PhD Bruno M Carvalho, PhD Jonathan Chambers, PhD Marta Cirah Pradas, MSc Orin Courtenay, PhD Shouro Dasgupta, PhD Matthew J Eckelman, PhD Zia Farooq, MSc Peter Fransson, PhD Elisa Gallo, PhD Olga Gasparyan, PhD Nube Gonzalez-Reviriego, PhD Prof Ian Hamilton, PhD Risto Hänninen, DSc Charles Hatfield, MUP Kehan He, PhD Aleksandra Kazmierczak, PhD Vladimir Kendrovski, PhD Harry Kennard, PhD Gregor Kiesewetter, PhD Rostislav Kouznetsov, PhD Hedi Katre Kriit, PhD Alba Llabrés-Brustenga, PhD Simon J Lloyd, PhD Martín Lotto Batista, MSc Carla Maia, PhD Prof Jaime Martinez-Urtaza, PhD Zhifu Mi, PhD Carles Milà, MSc Jan C Minx, PhD Prof Mark Nieuwenhuijsen, PhD Julia Palamarchuk, PhD Dafni Kalatzi Pantera, PhD Marcos Quijal-Zamorano, MSc Peter Rafaj, PhD Prof Elizabeth J Z Robinson, PhD Nacho Sánchez-Valdivia, MSc Daniel Scamman, EngD Oliver Schmoll, Dipl.-Ing Maquins Odhiambo Sewe, PhD Jodi D Sherman, MD Pratik Singh, MSc Elena Sirotkina, MA Henrik Sjödin, PhD Prof Mikhail Sofiev, PhD Balakrishnan Solaraju-Murali, PhD Prof Marco Springmann, PhD Marina Treskova, PhD Joaquin Triñanes, PhD Eline Vanuytrecht, PhD Fabian Wagner, PhD Maria Walawender, PhD Laura Warnecke, PhD Ran Zhang, MSc Marina Romanello, PhD Prof Josep M Antó, PhD Prof Maria Nilsson, PhD Prof Rachel Lowe,
Abstract
Record-breaking temperatures were recorded across the globe in 2023. Without climate action, adverse climate-related health impacts are expected to worsen worldwide, affecting billions of people. Temperatures in Europe are warming at twice the rate of the global average, threatening the health of populations across the continent and leading to unnecessary loss of life. The Lancet Countdown in Europe was established in 2021, to assess the health profile of climate change aiming to stimulate European social and political will to implement rapid health-responsive climate mitigation and adaptation actions. In 2022, the collaboration published its indicator report, tracking progress on health and climate change via 33 indicators and across five domains.
This new report tracks 42 indicators highlighting the negative impacts of climate change on human health, the delayed climate action of European countries, and the missed opportunities to protect or improve health with health-responsive climate action. The methods behind indicators presented in the 2022 report have been improved, and nine new indicators have been added, covering leishmaniasis, ticks, food security, health-care emissions, production and consumption-based emissions, clean energy investment, and scientific, political, and media engagement with climate and health. Considering that negative climate-related health impacts and the responsibility for climate change are not equal at the regional and global levels, this report also endeavours to reflect on aspects of inequality and justice by highlighting at-risk groups within Europe and Europe’s responsibility for the climate crisis.
Published in The Lancet
DOI: 10.1016/s2468-2667(24)00055-0
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Authors: Ceylan Polat, Nazli Ayhan, Koray Ergünay, Remi N. Charrel
Abstract
Sandfly-borne phleboviruses (SBPs), which cause sandfly fever, aseptic meningitis, encephalitis, and meningoencephalitis, are emerging pathogens of major public health concern. Virus nucleic acid testing is essential for SBP diagnosis, especially in the early stages of infection, and for the discovery of novel SBPs. The efficacy of utilizing generic primers that target conserved nucleotide sequences for the detection of both known and novel SBPs has not been extensively evaluated. We aimed to compare and evaluate the performance of five generic primer sets, widely used to detect S- and L-segments of arthropod-borne phleboviruses and designed as singleplex (n = 3) and nested (n = 2) formats, including both well-known and recently characterized 15 Old World virus strains. Furthermore, we performed in silico analysis to assess the detection capabilities of these generic primer sets. The initial evaluation of previously published generic primer sets for SBP detection yielded two singleplex primer sets with the potential to be adapted for use in real-time or high-throughput detection settings. Studies are ongoing to develop and further optimize a preliminary assay and test various hosts and vectors to assess their capacity to detect known and novel viruses.
IMPORTANCEVirus nucleic acid testing is the primary diagnostic method, particularly in the early stages of illness. Virus-specific or syndromic tests are widely used for this purpose. The use of generic primers has had a considerable impact on the discovery, identification, and detection of Old World sandfly-borne phleboviruses (OWSBP). The study is significant because it is the first to carry out a comparative evaluation of all published OWSBP generic primer sets.
Published in ASM/ Microbiology Spectrum
DOI: 10.1128/spectrum.03428-23
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Authors: Carla Maia
Abstract
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.
Published in Journal of Comparative Pathology
DOI: 10.1016/j.jcpa.2024.01.001
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Authors: Rafael Rocha, Cláudia Conceição, Luzia Gonçalves, Carla Maia
Abstract
Objective
To compare the current knowledge and practices regarding leishmaniasis among blood donors and health students and professionals, in Portugal.
Material and methods
Data were collected through the application of two questionnaires (one online and one in paper) with similar questions in two distinct cross-sectional independent studies, each targeting one of the groups. Descriptive statistics and hypothesis testing were performed using IBM® SPSS® Statistics.
Results
In total, 3763 blood donors, 254 students and 232 professionals were included in the comparative analysis. Over 95% of students and professionals, but only around 70% of blood donors had previously heard of leishmaniasis. Over 90% of participants in each group admitted leishmaniasis affected animals, but only in the professional group over 90% were aware of human leishmaniasis.
Conclusions
Even though canine leishmaniasis is recognized by many blood donors and by most students and professionals, awareness of the disease in humans is less common, highlighting the importance of promoting an approach to this infection through a One Health lens.
Published in One Health
DOI: 10.1016/j.onehlt.2024.100697
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Authors: J. Risueño , S. Bersihand , C. Bender , T. Cornen , K. De Boer , A. Ibáñez-Justicia , D. Rey , Y. Rozier , A. Schneider , A. Stroo , A. Vanslembrouck , W. Van Bortel , A. Weigand , D. Zambianchi , P. Pérez Cutillas , S. Oerther , M. Braks , G.R.W. Wint , E. Berriatua , and F. Schaffner
Abstract
Sand flies are vectors of Leishmania spp. protozoa, phleboviruses and Bartonella bacilliformis. In this study we surveyed areas of central-western Europe, encompassing the northern limit of the known sand fly distribution, and investigated the relationship between their presence and environmental variables. In this area, very limited occurrence data exists, none of them recent. Sampling was performed in July and August 2023 using CDC-light traps, sticky traps and human landing captures, at 179 selected sites in 48 municipalities. A total of 55 sand fly specimens were collected at 11% (20/179) of the sites sampled – including 16 sites in France, 2 in Luxembourg and 1 in Germany – comprising the first published records for Luxembourg and the Trier-Saarburg County of Germany. No sand flies were detected in Belgium or in the Netherlands. Two species were captured: Phlebotomus mascittii (37 females, 2 males) and Ph. perniciosus (1 female, 15 males). The latter species was only found in Savoie, the southernmost region sampled in France, while the former species was detected as far north as latitude 50°N. Logistic regression modelling indicated that the probability of sand fly presence gradually decreased with increasing latitude and altitude (P < 0.05), and it was not associated to other analysed landscape features in the proximity of the traps. The study confirms and provides new evidence for the presence of sand flies in areas of Western Europe and highlights the need to be aware of potential autochthonous transmission of sand fly-borne pathogens in these areas.
Published in European Mosquito Control Association
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Authors: Claudia Cosma, Carla Mai, Nushrat Khan, Maria Infantino and Marco Del Riccio
Abstract
Leishmaniasis is classified as a neglected tropical disease (NTD), caused by protozoan parasites of the genus Leishmania, which are transmitted to humans and other animals through the bite of infected female phlebotomine sandflies. There are three forms of the disease: cutaneous leishmaniasis (CL) manifested by ulcers and scars; systemic or visceral leishmaniasis (VL), which can lead to life-threatening complications if left untreated; and mucocutaneous leishmaniasis (MCL), which can destroy the mucous membranes of the nose, mouth and throat. Human leishmaniasis is endemic in many countries across Africa, Asia, Southern Europe, the Middle East, and Central and South America. The interconnection of environmental, animal and human health underlies the spread of the Leishmania parasite. Environmental disruptions, such as climate change, deforestation or urbanisation, but also globalisation and migration, significantly affect the distribution and abundance of sand fly vectors and reservoir hosts. Climate change alters the breeding patterns of sandflies and expands their geographic range; deforestation and misuse of large areas disrupt ecosystems, leading to increased human-vector contact; and urbanisation increases the potential for contact between parties, particularly in densely populated areas. Migration of humans and animals, either through natural migration or, for example, the pet trade and breeding, can facilitate the spread of Leishmania parasites. In addition, socio-economic factors, including poverty and lack of access to healthcare, increase the burden of leishmaniasis in vulnerable populations. Due to this multitude of reasons, the geographic distribution of sandflies has expanded to higher latitudes and altitudes in recent years, with a consequent increase in disease burden. Indeed, despite ongoing challenges in the surveillance systems, data from the last available year have shown an increase in many cases in both humans and dogs. This perspective explores the interconnected factors influencing the spread of leishmaniasis worldwide and the epidemiology of the disease. In addition, it illustrates the importance of integrated strategies in a One Health approach: surveillance, prevention and control of vectors, animals and humans.
DOI: 10.3390/tropicalmed9110258
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Authors: Nazli Ayhan, Carole Eldin, Remi Charrel
Abstract
Background: Toscana virus (TOSV) is a sand fly-borne phlebovirus causing central nervous system (CNS) infection in Mediterranean countries, during summer season. However, clinical aspects of the disease caused by this virus are poorly known by clinicians, so that its prevalence is probably underestimated due to a lack of diagnosis.
Study design: The data was gathered from all available case series and retrospective studies identifying TOSV as the causative viral agent. The informations of age, sex, clinical characteristics, laboratory findings, imaging results and clinical outcomes of TOSV infection were recorded and analyzed.
Results: A total of 95 articles including TOSV infections resulting in a total of 1381 cases, were analyzed. Our findings indicate that TOSV affects individuals across various age groups, with a median age of 44.45 years. A notable disparity in infection rates between genders, with men being significantly more likely to present symptoms due to TOSV than women, with a sex ratio of 2.0. The clinical presentation of TOSV infection encompasses a range of symptoms, including fever, headache, retro-orbital pain, neurological and muscular manifestations with less common reports of cutaneous and gastrointestinal symptoms. To date, six fatalities have been attributed to TOSV infections, with a median age of 76 years. Diagnostic evaluation of TOSV infections often involves the analysis of cerebrospinal fluid, where findings may include an elevated white blood cell count.
Conclusions: These findings underscore the diverse clinical manifestations of TOSV infections including flu like symtomps. TOSV is an emerging infectious threat that warrants inclusion in the diagnostic protocols for patients presenting with CNS, particularly within the Mediterranean basin or for those with recent travel history to endemic regions during warmer months when sand flies are actively circulating.
DOI: 10.1016/j.jinf.2025.106415
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Authors: Wilfrid Sewade, Ceylan Polat, Ozge Erisoz Kasap
Abstract
Background
Mosquitoes transmit various pathogens causing diseases like Zika, Dengue, West Nile and Chikungunya. They also harbour insect-specific viruses (ISVs) and Wolbachia, which can block arbovirus transmission. This study investigated the prevalence of Orthoflavivirus and Wolbachia in mosquito populations from three provinces in Türkiye.
Methods
Mosquitoes were collected using CDC Miniature Light traps in 2022–2023. Morphologically identified specimens were pooled (1–10 individuals) and screened for Orthoflavivirus and Wolbachia via PCR and confirmed by Sanger sequencing. Infection prevalence was estimated using the maximum likelihood method. Mosquito taxa richness across provinces was estimated using the abundance-based, non-parametric Chao1 index.
Results
Among 8766 mosquitoes (11 taxa) collected, Culex perexiguus, Ochlerotatus caspius and Anopheles claviger were most abundant. Anopheles flavivirus (AnFV) detected in one Oc. caspius pool, while Wolbachia sequences belonging to supergroup B were detected in An. claviger, Cx. pipiens s.l., Cx. perexiguus and Oc. caspius, with an overall infection prevalence of 0.0119 (95% CI: 0.008–0.0161). The richest mosquito fauna was detected in Ankara, followed by Adana, and Çankırı.
Conclusion
This study provides new insights into mosquito richness and the prevalence of Orthoflavivirus and Wolbachia in Türkiye, contributing to vector surveillance and the potential use of Wolbachia in mosquito control strategies.
DOI: https://doi.org/10.1111/tmi.14099
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Authors: Mert Okbay, Ozge Erisoz Kasap
Abstract
Vertebrate and plant hosts provide arthropods with a range of stimuli to aid in their localization, among which semiochemicals, volatile cues enabling hosts to be located through their odours, stand out as particularly important. Recognizing their potential in vector control, this study investigates semiochemicals as viable alternatives for effective vector management strategies. In this study, behavioral responses of Phlebotomus papatasi (Scopoli, 1786) adults to 11 semiochemicals, derived from plant and vertebrate hosts and previously tested on various vector species, across 3 concentrations (10-2%, 10-3%, and 10-4%) were evaluated using a dual-choice olfactometer. A total of 1,110 females and 1,110 males were individually assayed, with behavioral responses quantified through trap selection and response times. Notably, 1-octanol (10-2%) consistently exhibited the lowest attractiveness, whereas octanal, decanal, 1-hexanol, 1-octen-3-ol, ocimene, linalool oxide, and sulcatone elicited significant attractive effects, with pronounced sex-specific and concentration-based differences. Mixtures, formulated from the most attractive individual concentrations, revealed both synergistic and antagonistic interactions, underscoring the complexity of chemical interplay in mediating host-seeking behavior. Statistical models demonstrated significant interactions among semiochemical type, concentration, and sex, influencing both behavior and decision latency. This study marks the first investigation into the attractant effects of plant- and animal-derived semiochemicals on Ph. papatasi adults within an olfactometer system. The results are expected to provide critical insights into sand fly ecology and the epidemiology of sand fly-borne diseases while contributing to the development of integrated vector management strategies.
DOI: 10.1093/jme/tjaf065
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Authors: Nazli Ayhan, PhD , Léa Luciani, PharmD PhD , Laura Pezzi, PhD , Jean Sélim Driouich, PharmD PhD , Raphaelle Klitting, PhD , Elif Kurum, MS , Nadim Cassir, MD PhD , Sophie Bayle, MD , Amandine Chollet, MD , Guillaume A Durand, MD PhD , Anne Darmon, MD , Julie Allemand, MD , Fabrice Edorh, MD , Laetitia Ninove, PharmD PhD , Antoine Nougairède, PharmD PhD , Gilda Grard, PhD , Xavier de Lamballerie, MD PhD , Rémi N Charrel, MD PhD
Abstract
This study confirms the continued circulation of Toscana virus (TOSV) lineage B in France, with nine laboratory-confirmed autochthonous neuroinvasive cases identified between 2022 and 2024. All patients presented with central nervous system symptoms, predominantly during summer months, and most were confirmed by RT-qPCR on cerebrospinal fluid. Phylogenetic analysis revealed close similarity to previously circulating French strains, supporting long-term local transmission. One case in eastern France raises concern for possible geographic expansion. These findings underscore the need for sustained surveillance and preventive measures in endemic regions, particularly for travellers and clinicians encountering summer-onset neurological syndromes.
DOI: 10.1093/jtm/taaf057
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Authors: Vladimir Ivović, Peter Glasnović, Sara Zupan, Tea Knapič, Tomi Trilar, Miša Korva, Nataša Knap, Urška Glinšek Biškup, Tatjana Avšič-Županc, Katja Adam
Abstract
Sandflies (Diptera: Psychodidae: Phlebotominae) are important vectors of pathogens, including Leishmania parasites and phleboviruses, but their distribution and seasonal activity in Slovenia have not been sufficiently studied. This study presents a comprehensive three-year (2020-2022) surveillance programme aimed at assessing the diversity of sandfly species, their distribution, seasonal dynamics and potential role as vectors of pathogens. A total of 1,240 sandflies were collected at 43 sampling sites across Slovenia, identifying Phlebotomus papatasi, P. neglectus, P. perniciosus and P. mascittii. The highest abundance and species diversity were observed in the Mediterranean and Karst regions. Seasonal activity peaked in July, with population fluctuations influenced by climatic conditions. Molecular analyses for Leishmania parasites and phleboviruses showed no positive results, indicating a low prevalence of pathogens in the sampled populations. Predictive habitat models indicate that environmental factors, particularly temperature and precipitation, play a decisive role in the spread of sandflies. While P. mascittii has the largest ecological range, its vector competence remains uncertain. The results provide important insights into the ecology of sandflies in Slovenia and emphasize the need for continuous surveillance in the context of climate change and emerging vector-borne disease risks.
DOI: 10.3389/fvets.2025.1603358
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Authors: Danyang Wang, Anouschka R. Hof, Kevin D. Matson, CLIMOS data providers Frank van Langevelde
Abstract
Climate and land-use changes influence the transmission of vector-borne diseases by affecting the distribution and survival of disease vectors. Numerous diseases are transmitted by phlebotomine sand flies (Diptera: Psychodidae: Phlebotominae), including leishmaniasis. Several major sand fly-borne diseases are responsible for high global disease burdens and high socio-economic costs. In Europe, over 20 known sand fly vector species are largely confined to the Mediterranean Basin, yet records of sand fly presence further north increase. Global warming is predicted to drive the spread of sand flies to large areas of Europe in the 21th century, an effect likely to be exacerbated by anthropogenic factors. However, the constraints to the geographic distributions of sand flies are not well understood. This study aims to increase the understanding of the drivers of the geographic distributions of sand flies, using species distribution modelling to systematically test links between sand fly occurrences and climatic, land-use, lithological, biodiversity and human population variables in Europe and adjacent Mediterranean regions. We found that moisture is the most important environmental variable both in explaining and in predicting sand fly occurrences. The projected suitable habitats are larger than the current known sand fly distributions, and these habitats are expected to expand due to changes in climate and land-use.