Sandfly-Borne Phleboviruses in Portugal: Four and Still Counting

ReviewThis article belongs to the Special Issue Sand Fly-Borne Phleboviruses, Volume II.According to ICTV, there are currently 66 known phlebovirus species. More than 40 of these viruses were isolated or detected in phlebotomine sandflies and some of them are known pathogens. In Portugal, information about sandfly-borne phleboviruses is scarce and scattered sandfly-borne diseases are neglected and often not considered in differential diagnoses. The main objective of this work was to gather the existing information and to raise awareness about the circulating phleboviruses in this country. To date, Massilia and Alcube phleboviruses have been isolated from sandflies in southern Portugal. Human infections with Toscana and Sicilian phleboviruses have been reported, as well as seroprevalence in cats and dogs. More studies are needed in order to understand if the viruses isolated during the entomological surveys have an impact on human health and to fully understand the real importance of the already recognized pathogens in our country.This research was supported in part by the Portuguese Foundation for Science and Technology (FCT) through the project: “Phleboviruses in Portugal: vectors, pathogenesis and co-infections” (PTDC/DTP-SAP/0859/2014).info:eu-repo/semantics/publishedVersio

Estabelecimento de prioridades para a conservação dos vertebrados terrestres do Parque Natural da Ria Formosa e selecção de áreas para a sua protecção

Dissertação de mest. em Gestão e Conservação da Natureza, Faculdade de Ciências do Mar e do Ambiente, Univ. do Algarve, 2004A biodiversidade encontra-se actualmente ameaçada em todo o mundo e nem mesmo nas áreas destinadas para a conservação é possível salvaguardar todas as espécies presentes. Assim, é necessário encontrar-se prioridades para a conservação. O objectivo deste trabalho consistiu na identificação das espécies de vertebrados prioritárias para a conservação no Parque Natural da Ria Formosa (uma área protegida no sul de Portugal Continental), e na definição de áreas para a sua preservação. Para tal foi feita uma ordenação das espécies baseada nos estatutos de conservação aos níveis nacional e internacional, na distribuição e sensibilidade ecológica de cada uma delas. As áreas para a conservação foram definidas através de algoritmos de selecção iterativos, baseados nos princípios da raridade e da complementaridade contidos no ResNet, um pacote de software disponível na internet. Das 26 espécies de vertebrados definidas como prioritárias incluem-se 19 de aves, três de répteis, três de anfíbios e uma de mamíferos. A espécie que maior valor obteve na ordenação foi a galinha-sultana, o símbolo do Parque Natural da Ria Formosa. Concluiu-se que as áreas seleccionadas para a protecção das espécies prioritárias englobam uma tal variedade de biótopos que se os esforços de conservação lhes forem direccionados, muitas outras espécies ficarão protegidas. Por fim, foram sugeridas medidas para a conservação da biodiversidade no PNRF.Biodiversity is currently threatened ali over the world and it is not possible to preserve ali species, even in areas that are set aside for nature conservation. Thus it is necessary to find conservation priorities. The goal of this work consisted on the identification of conservation priorities for the vertebrate species in the Natural Park of Ria Formosa (a protected area in the South of Continental Portugal), and in the definition of areas for their preservation. The species priorization was based on their conservation status at national and intemational leveis, as well as on their distribution and ecological sensitivity. The conservation areas had been defined through iterative selecction algorithms, based in the principies of rarity and complementarity displayed in ResNet, a software package available in the Internet. From the 26 vertebrate chosen species, 19 were birds, three were reptiles, three were amphibians and one was a mammal. The species that obtained a higher value in the rank was Porphyrio porphyrio, which is the symbol of Natural Park of Ria Formosa. It was concluded that the selected areas for the proteccion of prioritary species contain one such variety of biotopes that if the conservation efforts will be directioned to them, many other species will be protected. Finally, conservation measures for biodiversity conservation in the Natural Park of Ria Formosa have been suggested

Phlebovirus, importância em saúde publica em Portugal

Tese de doutoramento, Biologia (Microbiologia), Universidade de Lisboa, Faculdade de Ciências, 2010O vírus Toscana (género Phlebovirus, família Bunyaviridae) é um vírus neurotrópico que circula na Bacia do Mediterrâneo. Embora o reservatório na natureza seja desconhecido, está confirmada a sua transmissão por dípteros do género Phlebotomus. Recentemente foram também detectados vírus do género Flavivirus (família Flaviviridae) nestes vectores. O objectivo principal desta tese foi determinar a importância dos flebovírus em Saúde Pública em Portugal. Deste modo, no âmbito deste trabalho foram padronizadas e implementadas técnicas para o diagnóstico laboratorial de flebovírus, tendo sido desenvolvida uma técnica de imunofluorescência indirecta (IFI) e implementada uma técnica de Enzyme Linked Immunosorbent Assay (ELISA), como teste confirmatório, para o diagnóstico de vírus Toscana. O índice de concordância entre as duas técnicas foi considerado excelente, sendo a sensibilidade da IFI de 100%, e a especificidade superior a 80% na detecção dos anticorpos pesquisados (IgG e IgM). Inquéritos sero-epidemiológicos realizados na população humana para detectar a presença de anticorpos anti-vírus Toscana incluíram 924 indivíduos, distribuídos por três populações. Na população considerada de risco a seroprevalência foi 3,4% e na população controlo foi 2%. Em relação à população de indivíduos com sintomatologia e pedido de diagnóstico laboratorial, a seroprevalência mais elevada foi detectada no grupo de indivíduos com sintomas do sistema nervoso central (4,2%), na qual foram confirmados cinco casos de infecção recente (3%). No grupo de indivíduos com sintomatologia não neurológica a seroprevalência foi 1,3%. No total de 29 soros com anticorpos anti-vírus Toscana foram confirmados cinco por testes de neutralização por redução em placas com a estirpe ISS.Phl.3, demonstrando-se a presença de mais do que um genótipo de vírus Toscana em circulação em Portugal. No intuito de determinar os prováveis reservatórios de flebovírus na natureza foram realizados estudos serológicos em vertebrados selvagens e domésticos, nomeadamente em roedores e canídeos selvagens e domésticos. Um lobo e uma raposa apresentaram anticorpos IgG. Os resultados sugerem que, provavelmente, os canídeos selvagens têm um papel importante na manutenção do ciclo biológico dos flebovírus na natureza. Para identificar as espécies de flebótomos em circulação em Portugal procedeu-se à captura de flebótomos com armadilhas CDC em duas zonas do país (Algarve e Arrábida). Foram capturados 7578 flebótomos, designadamente 3330 machos e 4248 fêmeas. A espécie P. perniciosus apresentou uma distribuição geográfica mais ampla e o maior efectivo populacional. Além de P. perniciosus, foram encontradas P. ariasi, P. sergenti e Sergentomyia minuta. Os resultados indicam que P. sergenti se encontra, provavelmente, em expansão no nosso país, já que ocorreu em altitudes baixas Sergentomyia minuta, foi encontrada no Algarve a partir dos 11 metros de altitude, o que indica que a sua distribuição também está a alterar-se. A identificação dos vírus em circulação nos vectores foi baseada no método de nestedRT-PCR. A presença de flebovírus não foi detectada 269 pools de flebótomos pesquisados. No entanto, num pool em 90 estudados foi revelada, pela primeira vez em Portugal, a presença de flavivírus em flebótomos. A sequência obtida é idêntica às sequências de flavivírus detectadas em mosquitos do género Culex no nosso país e apresenta 98% de identidade com a sequência de um flavivírus encontrado em Ochlerotatus caspius em Itália. A circulação de flebovírus em Portugal foi comprovada nos seres humanos e em potenciais reservatórios. Foi demonstrada, pela primeira no nosso país, a circulação de flavivírus em flebótomos. Os resultados obtidos justificam a continuação da investigação na área dos vírus transmitidos por flebótomos pela sua importância em Saúde Pública. Flebovírus, vírus Toscana, flavivírus, flebótomos, reservatóriosToscana virus (Phlebovirus genus, family Bunyaviridae) is a neurotropic virus which circulates in the Mediterranean Basin. Although the reservoir in nature is unknown, it is a fact that this virus is transmitted by sandflies of Phlebotomus genus. Viruses of the genus Flavivirus (family Flaviviridae) were recently detected in these vectors. The main objective of this thesis was to determine the importance of Phleboviruses in Public Health in Portugal. Within the aim of this work an in-house indirect immunofluorescence assay (IFI) was developed and an enzyme-linked immunosorbent assay (ELISA) with a commercial kit was also implemented for the laboratory diagnosis of Toscana virus. The agreement index between the two techniques was excellent, the sensitivity of IFI was 100% and the specificity exceeded 80% for both antibodies (IgG and IgM). Sero-epidemiological surveys were conducted in the human population, in 924 individuals, distributed in three populations, for the detection of antibodies anti Toscana virus. In the population considered at risk the prevalence was 3.4% and in the control population was 2%. In what concerns to symptomatic individuals with request for laboratorial diagnosis, the highest seroprevalence was detected in the population of individuals with neurological symptoms (4.2%), were there were five confirmed cases of recent infection (3%). In the population of individuals with no neurological symptoms, the seroprevalence was 1.3%. Five of the 29 positive sera were confirmed by plaque reduction neutralization tests with the strain ISS.Phl.3. This shows the presence of more than one genotype of Toscana virus circulating in Portugal. To determine the probable phleboviruses reservoirs in nature serological studies were performed on 538 samples of wild and domestic vertebrates, including rodents and wild canids. IgG antibodies were detected in two animals, namely a wolf and a fox. Toscana virus (Phlebovirus genus, family Bunyaviridae) is a neurotropic virus which circulates in the Mediterranean Basin. Although the reservoir in nature is unknown, it is a fact that this virus is transmitted by sandflies of Phlebotomus genus. Viruses of the genus Flavivirus (family Flaviviridae) were recently detected in these vectors. The main objective of this thesis was to determine the importance of Phleboviruses in Public Health in Portugal. Within the aim of this work an in-house indirect immunofluorescence assay (IFI) was developed and an enzyme-linked immunosorbent assay (ELISA) with a commercial kit was also implemented for the laboratory diagnosis of Toscana virus. The agreement index between the two techniques was excellent, the sensitivity of IFI was 100% and the specificity exceeded 80% for both antibodies (IgG and IgM). Sero-epidemiological surveys were conducted in the human population, in 924 individuals, distributed in three populations, for the detection of antibodies anti Toscana virus. In the population considered at risk the prevalence was 3.4% and in the control population was 2%. In what concerns to symptomatic individuals with request for laboratorial diagnosis, the highest seroprevalence was detected in the population of individuals with neurological symptoms (4.2%), were there were five confirmed cases of recent infection (3%). In the population of individuals with no neurological symptoms, the seroprevalence was 1.3%. Five of the 29 positive sera were confirmed by plaque reduction neutralization tests with the strain ISS.Phl.3. This shows the presence of more than one genotype of Toscana virus circulating in Portugal. To determine the probable phleboviruses reservoirs in nature serological studies were performed on 538 samples of wild and domestic vertebrates, including rodents and wild canids. IgG antibodies were detected in two animals, namely a wolf and a fox.The results suggest that probably wild canids have an important role in maintaining the biological cycle of phleboviruses in nature. For the identification of sandfly species in circulation in Portugal, captures of these vectors were performed with CDC light traps, in two areas of the country, namely Algarve and Arrabida. In the two prospection periods 7578 sandflies were captured, of which 3330 were male and 4248 were females. The species P. perniciosus showed a wider distribution and the greater effective population size. The obtained results during this work indicate that the geographic distribution of P. Sergenti is getting wider since it has occurred at lower altitudes. Sergentomyia minuta was also found at lower altitudes then previously expected, since it was present in the Algarve in an altitude as low as 11 meters. The identification of circulating virus in the vectors was based on nested RT-PCR method. The presence of phleboviruses was not detected in 269 screened pools. However, the presence of Flavivirus genus in sandflies was detected, for the first time in Portugal, in one of 90 pools surveyed. The obtained sequence is identical to the sequences of flaviviruses detected in mosquitoes of the genus Culex in our country and shows 98% identity with the sequence of a flaviviruses found in Ochlerotatus caspius in Italy. The circulation of phleboviruses in Portugal was proven in humans and in potential reservoirs. The presence of flaviviruses in sandflies was revealed for the first time in Portugal. These results justify the continuation of research on viruses transmitted by sandflies due to their importance in what concerns to Public health. Phleboviruses, Toscana virus, flaviviruses, sandflies, reservoir

Parcerias estratégicas público-privadas no sistema rodoviário português; o caso das SCUT: diagnóstico e perspectivas

Diante dos desafios que as estratégias de desenvolvimento económico e das Politicas Públicas do setor rodoviário colocaram, a Administração Rodoviária adapta, na década de 90, novas formas de Gestão Pública, com uma nova cultura e novos modelos organizacionais, assumindo uma reengenharia financeira capaz de adotar novos modelos de financiamento rodoviário. Os novos modelos de financiamento basearam-se nos modelos desenhados do Project Finance sobre a forma de Parcerias Público-Privadas (PPP), que apareceram como a solução do Estado, para implementar a sua Estratégia no setor e ao mesmo tempo não deixar de cumprir as limitações orçamentais impostas pela adesão ao Euro. Com este trabalho pretende-se identificar os diferentes modelos de financiamento do setor rodoviário e diagnosticar o contexto organizacional que os enquadra, discutir a celebração das PPP e, como Estudo de Caso, analisar as Concessões SCUT (sem custo para o utente) nas suas diferentes vertentes, perspetivando o seu futuro; ABSTRACT: Given the challenges that economic development strategies and public policies of the road sector put, the Road Administration adopts in the 90's, new forms of Public Management, with a new culture and new organizational models, assuming a financial reengineering able to adopt new models for road funding. The new models of funding were based on models drawn from the Project Finance on the Public- Private Partnerships (PPP), which appeared as the solution of the state to implement its strategy in the road sector while not failing to reach budgetary constraints imposed by rules of the “Euro”. This work aims to identify the different types of road sector financing models and diagnose their organizational context framework, to discuss the celebration of the PPP and as a Case Study, and to analyze different aspects of “SCUT” Concessions (no cost to the user) and its future prospects

Vírus Toscana: dez anos de diagnóstico em Portugal

[EN] Introduction: Toscana virus (TOSV) is an emerging sandfly-borne virus within the Phlebovirus genus. Although most infections caused by this virus present as asymptomatic or with minimal symptomatology, TOSV may emerge as a febrile disease or sporadic cases of neurological disease such as meningitis or meningoencephalitis. This pathogen is distributed throughout the Mediterranean basin, along with the spatial distribution of its recognized sandfly vector, Phlebotomus perniciosus. Portugal, after Italy, was the second country considered endemic for this virus, with the first case of acquired infection published in 1985. Although little is known about the circulation of this virus in Portugal, the laboratory diagnosis of TOSV is available at the Centre for Vectors and Infectious Diseases Research of the National Institute of Health Dr. Ricardo Jorge (CEVDI/INSA), since 2007. The aim of this study is to report the results of the diagnosis of TOSV at the CEVDI/INSA, between 2009 and 2018. Material and Methods: The diagnosis of TOSV in the CEVDI/INSA is included in the arboviruses and vector-borne neurotropic viruses panels or can be performed, when specified, for TOSV only. Direct detection is made in cerebrospinal fluid samples and is available for TOSV by specific real-time reverse transcription polymerase chain reaction followed by conventional real-time reverse transcription polymerase chain reaction for sequencing purposes, if positive. For indirect diagnosis, performed in serum samples, an in-house immunofluorescence assay for the detection of IgM and IgG antibodies against TOSV is used. A commercial immunofluorescence assay consisting in a mosaic of four phleboviruses is also available, in which, in addition to TOSV, antibody detection for sandfly fever Naples virus, sandfly fever Sicilian virus and sandfly fever Cyprus virus can be done. All diagnostic tests requested by clinicians to the CEVDI/ INSA for arboviruses, neurotropic viruses and/or TOSV between January 2009 and December 2018, were included in this study. Results: During the study period, the CEVDI/INSA received samples from 608 patients with diagnostic requests for TOSV. Five acute TOSV infections and one acute sandfly fever Sicilian virus infection were confirmed in serum samples. Three other patients had serological evidence of previous contact with the virus. Two of the six patients with acute infection developed febrile syndrome, and the other four presented with neurological disease: meningitis (n = 2), meningoencephalitis (n = 1) and severe depression of consciousness (n = 1). These infections were most likely acquired in the districts of Faro (3), Lisbon (2) and Setúbal (1). Discussion: In Portugal, the number of laboratory diagnostic requests for TOSV is low when compared to the number of requests for other less prevalent vector-borne viruses. The Faro district presented the highest number of TOSV-specific diagnostic requests which seems to indicate a higher level of recognition by clinicians in that region. Febrile syndrome and neurological disease were the clinical manifestations that were present in acute cases. In this study, in addition to the Faro district, recent infections were also detected in the districts of Lisbon and Setúbal. It is probable that TOSV may be distributed throughout the mainland territory since its main vector is present from north to south. In 2017, the sandfly fever Sicilian virus was associated for the first time with human disease in our country, thus alerting to the circulation of this phlebovirus. Conclusion: Even though the number of identified cases in Portugal is low, TOSV circulates and causes disease in our country. The diagnosis of this and other phleboviruses should not be neglected in the differential diagnosis of febrile syndrome and viral meningitis and meningoencephalitis, especially during the warmer months, when the vector’s activity is higher.[PT] Introdução: O vírus Toscana (TOSV) é um vírus emergente, transmitido por flebótomos, que pertence ao género Phlebovirus. Apesar de a maioria das infeções causadas por este vírus serem assintomáticas ou apresentarem uma sintomatologia ligeira, o TOSV pode causar síndrome febril ou casos esporádicos de doença neurológica tais como meningite ou meningoencefalite. Este agente patogénico encontra-se distribuído por toda a bacia do Mediterrâneo, de acordo com as áreas de distribuição do seu vetor reconhecido, Phlebotomus perniciosus. Depois de Itália, Portugal foi o segundo país considerado endémico para este vírus após a publicação, em 1985, do primeiro caso de infeção adquirida no nosso território. Apesar do pouco conhecimento acerca da circulação deste vírus, no nosso país, o diagnóstico laboratorial de TOSV está disponível em Portugal, desde 2007, no Centro de Estudos de Vetores e Doenças Infeciosas do Instituto Nacional de Saúde Dr. Ricardo Jorge (CEVDI/INSA). O objetivo deste trabalho é dar a conhecer os resultados do diagnóstico de TOSV em Portugal, de 2009 a 2018, no CEVDI/INSA. Material e Métodos: O diagnóstico de TOSV no CEVDI/INSA está inserido nos painéis ‘arbovírus’ e ‘vírus neurotrópicos transmitidos por vetores’ ou pode ser realizado, quando especificado, só para TOSV. O diagnóstico direto é realizado em amostras de líquido cefalorraquidiano e encontra-se disponível no CEVDI/INSA por RT-PCR em tempo real, específico para TOSV, seguido de RT-PCR convencional, no caso de a amostra ser positiva na primeira técnica, para confirmação por sequenciação. Para o diagnóstico indireto, realizado em amostras de soro, é utilizado uma técnica de imunofluorescência in-house, para a deteção de anticorpos IgM e IgG anti-TOSV. Também está disponível uma imunofluorescência comercial, com um mosaico de quatro flebovírus, onde para além do TOSV, são testados anticorpos contra três vírus da febre por flebótomos, nomeadamente Nápoles, Sicília e Chipre. Neste trabalho foram considerados os pedidos de diagnóstico ao CEVDI/INSA para arbovírus, vírus neurotrópicos e/ou TOSV, de janeiro de 2009 a dezembro de 2018. Resultados: No período em estudo, foram enviadas ao CEVDI/INSA, amostras de 608 indivíduos com pedido de diagnóstico de TOSV. Foram confirmadas cinco infeções agudas por TOSV e uma infeção aguda por vírus Sicília em amostras de soro. Três outros doentes apresentaram prova serológica de contacto prévio com o TOSV. Dois dos doentes com infeção aguda apresentaram síndrome febril, mas quatro evidenciaram quadros neurológicos: meningite (n = 2), meningoencefalite (n = 1) e alterações graves do estado de consciência (n = 1). Estas infeções foram, muito provavelmente, adquiridas nos distritos de Faro (3), Lisboa (2) e Setúbal (1). Discussão: Em Portugal, o número de pedidos de diagnóstico laboratorial para TOSV é baixo quando comparado com o número de pedidos para outros vírus transmitidos por vetores. O distrito de Faro foi o que apresentou o número mais alto de pedidos de diagnóstico específicos para TOSV, o que parece demonstrar que existe um maior reconhecimento pelos clínicos daquela região. Síndrome febril e doença neurológica foram as manifestações clínicas nos casos agudos. Neste estudo, além do distrito de Faro, foram também detetadas infeções recentes nos distritos de Lisboa e Setúbal. É provável que o TOSV se encontre distribuído por todo o território continental, uma vez que o seu principal vetor está presente de norte a sul. Em 2017, o vírus Sicília foi associado, pela primeira vez, a doença humana no nosso país, alertando para a circulação deste flebovírus. Conclusão: Apesar do número de casos identificados em Portugal ser baixo, o TOSV circula e causa doença no nosso país. Este e outros flebovírus não deveriam ser negligenciados no diagnóstico diferencial de síndrome febril e de meningites e meningoencefalites virais, em especial nos meses mais quentes, quando é maior a atividade dos seus vetores.This work was partially supported by the FCT Project: Phleboviruses in Portugal: vectors pathogenesis and coinfections (PTDC/DTP-SAP/0859/2014).info:eu-repo/semantics/publishedVersio

Remote monitoring of disease vector mosquitoes with a new optical sensor system for automatic classification

Introduction: - Mosquitoes represent a major threat to public health given their ability to transmit several pathogens. Some species of Aedes can transmit viruses such as dengue, Zika, or chikungunya. - Prevention of vector-borne diseases largely depends on effective and sustainable vector surveillance. Objectives: - Help to develop a novel bioacustic sensor that is able to identify the mosquitoes' species in real time - Deploy the sensor in the field in Madeira and Algarve.N/

Serological evidence of Toscana virus infection in Portuguese patients.

SUMMARYToscana virus (TOSV) is an emergingPhlebovirusof growing interest as a human pathogen in the Mediterranean Basin. In Portugal, however, little is known about the prevalence of TOSV infection. The aim of this work was to perform a seroprevalence study in patients with requests for laboratory diagnosis of vector-borne viruses. A total of 538 patients with and without neurological signs from 2004 to 2008 were studied by in-house indirect immunofluorescence assay and commercial enzyme-linked immunosorbent assays. A prevalence of 4·2% for IgG antibodies was found in the group of patients with neurological signs. Five (3%) of these had recent infections. In the group with no neurological signs, the IgG prevalence was 1·3%. Two samples, belonging to two patients, were also confirmed with plaque reduction neutralization tests with the TOSV ISS. Phl.3 Italian strain. This work showed that TOSV is present and causing disease from north to south in Portugal. The probable circulation of different phlebovirus serotypes in Portugal emphasizes the need for further studies

Phylogenetic analysis of Massilia phlebovirus in Portugal

© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).In the last two decades, molecular surveys of arboviruses have enabled the identification of several new viruses, contributing to the knowledge of viral diversity and providing important epidemiological data regarding possible new emerging viruses. A combination of diagnostic assays, Illumina sequencing and phylogenetic inference are here used to characterize two new Massilia phlebovirus strains isolated from sandflies collected in the Arrábida region, Portugal. Whole genome sequence analysis enabled their identification as reassortants and the recognition of genomic variants co-circulating in Portugal. Much is still unknown about the life cycle, geographic range, evolutionary forces and public health importance of these viruses in Portugal and elsewhere, and more studies are needed.This research was supported in part by the Portuguese Foundation for Science and Technology (FCT) through the project “Phleboviruses in Portugal: vectors, pathogenesis and co-infections” (PTDC/DTP-SAP/0859/2014). J.L. was supported by a research lectureship by the Zoology Department of the University of Oxford. M.G. was supported by Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ). This manuscript received the support of Fundação para a Ciência e a Tecnologia, grant number UIDB/04295/2020 and UIDP/04295/2020.info:eu-repo/semantics/publishedVersio

Co-Circulation of Leishmania Parasites and Phleboviruses in a Population of Sand Flies Collected in the South of Portugal

This article belongs to the Special Issue Leishmaniasis: Vector-Host-Pathogen Interactions in Health and Disease.In the Old World, phlebotomine sand flies from the genus Phlebotomus are implicated in the transmission of Leishmania spp. parasites (Kinetoplastida: Trypanosomatidae) and viruses belonging to the genus Phlebovirus (Bunyavirales: Phenuiviridae). Two of the five sand fly species known to occur in Portugal, Phlebotomus perniciosus and Ph. ariasi, the former being the most ubiquitous, are recognized vectors of Leishmania infantum, which causes visceral leishmaniasis, the most prevalent form of leishmaniasis in the country. Phlebotomus perniciosus is also the vector of the neurotropic Toscana virus, which can cause aseptic meningitis. Entomological surveillance is essential to provide fundamental data about the presence of vectors and the pathogens they can carry. As such, and given the lack of data in Portugal, an entomological survey took place in the Algarve, the southernmost region of the country, from May to October 2018. Polymerase chain reaction assays were performed in order to detect the presence of the above-mentioned pathogens in sand fly pools. Not only were both Leishmania parasites and phleboviruses detected during this study, but more importantly, it was the first time their co-circulation was verified in the same sand fly population collected in Portugal.This research was partially supported by the Portuguese Foundation for Science and Technology (FCT) through the project: “Phleboviruses in Portugal—vectors, pathogenesis and co-infections” (PTDC/DTP-SAP/0859/2014).info:eu-repo/semantics/publishedVersio