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Most human infections are asymptomatic or cause influenza-like illness; however, TOSV can cause severe neurological disease. Within the framework of the European Climate Monitoring and Decision Support Framework for Sand Fly\u2013borne Diseases (CLIMOS) project, this study reports the molecular detection of sand\u00a0fly-borne phleboviruses over 2 years of entomological surveillance in southern Portugal, aiming to characterize their genetic diversity and geographic distribution.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Phlebotomine sand\u00a0flies were collected using Centers for Disease and Control (CDC) miniature light traps between April and November of 2023 and 2024 in different regions of southern Portugal, including the Lisbon Metropolitan Area and the Algarve region. Additional collections were carried out in the Portalegre district between June and October 2024. After morphological identification, specimens were pooled according to species, sex, collection date, and location. Engorged females were analyzed individually. Molecular screening included a conventional polymerase chain reaction (PCR) for pan-\n                      <jats:italic>Phlebovirus<\/jats:italic>\n                      detection and a real-time reverse transcription quantitative (RT-q)PCR for the specific detection of TOSV and SFSV.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      A total of 7719 sand\u00a0flies were collected\u20144131 in 2023 (620 pools and 305 engorged females) and 3588 in 2024 (1062 pools and 393 engorged females).\n                      <jats:italic>Phlebotomus perniciosus<\/jats:italic>\n                      was the dominant species in both years and across all regions. Other identified species included\n                      <jats:italic>Phlebotomus ariasi<\/jats:italic>\n                      ,\n                      <jats:italic>Phlebotomus sergenti<\/jats:italic>\n                      ,\n                      <jats:italic>Phlebotomus papatasi<\/jats:italic>\n                      , and\n                      <jats:italic>Sergentomyia minuta<\/jats:italic>\n                      . Four\n                      <jats:italic>Phlebovirus<\/jats:italic>\n                      -positive pools were identified, all from\n                      <jats:italic>P.\u00a0perniciosus<\/jats:italic>\n                      collected in the Algarve region\u2014two in 2023 and two in 2024, including TOSV (\n                      <jats:italic>n<\/jats:italic>\n                      \u2009=\u20091), Alcube virus (\n                      <jats:italic>n<\/jats:italic>\n                      \u2009=\u20091),\n                      <jats:italic>Phlebovirus<\/jats:italic>\n                      strain PoSFPhlebV\/21\/2007 related to Massilia virus (\n                      <jats:italic>n<\/jats:italic>\n                      \u2009=\u20091), and Arrabida virus (\n                      <jats:italic>n<\/jats:italic>\n                      \u2009=\u20091) accordingly partial L segment sequence analysis.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      This study highlights the diversity and ongoing co-circulation of sand\u00a0fly-borne phleboviruses in southern Portugal, emphasizing the need for surveillance and diagnostic efforts to encompass a broader range of\n                      <jats:italic>Phlebovirus<\/jats:italic>\n                      species. The consistent association of\n                      <jats:italic>P.\u00a0perniciosus<\/jats:italic>\n                      with multiple viral species and locations reinforces its role as a key vector, warranting targeted vector-management strategies and inclusion in risk-prediction models.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s13071-026-07394-1","type":"journal-article","created":{"date-parts":[[2026,4,19]],"date-time":"2026-04-19T10:05:04Z","timestamp":1776593104000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Catching up with old friends: a 2-year survey of phlebotomine sand\u00a0fly-borne phleboviruses presence in southern Portugal"],"prefix":"10.1186","volume":"19","author":[{"given":"Carla","family":"Maia","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ricardo","family":"Parreira","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elif","family":"Kurum","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Federico","family":"Lucchese","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jo\u00e3o","family":"Cruz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rafael","family":"Marm\u00e9","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yasmina","family":"Martinez-Barciela","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alejandro","family":"Polina","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R\u00e9mi","family":"Charrel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nazli","family":"Ayhan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,4,19]]},"reference":[{"key":"7394_CR1","doi-asserted-by":"publisher","DOI":"10.3390\/v13050918","volume":"13","author":"CH Calisher","year":"2021","unstructured":"Calisher CH, Calzolari M. 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Owners of the properties signed an informed consent form allowing the setting of the CDC traps in their lands for this study.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"The authors declare no competing interests.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}},{"value":"During the preparation of this work the authors used Perplexity AI tool to improve the readability and language of the manuscript. After using this tool, the authors reviewed and edited the content as needed and take full responsibility for the content of the published article.","order":5,"name":"Ethics","group":{"name":"EthicsHeading","label":"Generative AI and AI-assisted technologies in the writing process"}}],"article-number":"236"}}