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Sand flies transmit Leishmania infantum, which is responsible for causing leishmaniasis, as well as many phleboviruses, including the human pathogenic Toscana virus. We screened sand flies collected from a single site between 2017 and 2020 for the presence of both phleboviruses and Leishmania. The sand flies were sampled with attractive carbon dioxide traps and CDC light traps between May and October. We collected more than 50,000 sand flies; 2826 were identified at the species level as Phlebotomus perfiliewi (98%) or Phlebotomus perniciosus (2%). A total of 16,789 sand flies were tested in 355 pools, and phleboviruses were found in 61 pools (6 Toscana virus positive pools, 2 Corfou virus positive pools, 42 Fermo virus positive pools, and 7 Ponticelli virus positive pools, and 4 unidentified phlebovirus positive pools). Leishmania was found in 75 pools and both microorganisms were detected in 16 pools. We isolated nine phleboviruses from another 2960 sand flies (five Ponticelli viruses and for Fermo viruses), not tested for Leishmania; the complete genome of a Fermo virus isolate was sequenced. The simultaneous detection in space and time of the Fermo virus and L. infantum is evidence that supports the co-circulation of both microorganisms in the same location and partial overlap of their cycles. A detailed characterization of the epidemiology of these microorganisms will support measures to limit their transmission. [ABSTRACT FROM AUTHOR]

Abstract The sand flies are the biological vectors of a variety of viruses belonging to the Phlebovirus genus. As several of these viruses, like Toscana virus, are important agents of diseases in humans, the definition the phleboviruses circulating in a particular area is an important health issue. This work presents results obtained between 2013 and 2016, by testing 25,853 field-collected sand flies, sampled at 50 sites in the Northern edge of the Apennine Mountains, in Emilia-Romagna and Lombardy regions (Italy). Isolation of the three reassortant Ponticelli I, II, and III viruses was successful from five pools of sand flies. These results, and phylogenetic analysis made by obtained sequences, suggest the presence of eight different viruses: Toscana virus, Fermo-like virus, Corfou virus, Ponticelli viruses, and two unknown putative viruses. The co-circulation of different phlebovirus reported in this study, indicate a very dynamic and complex situation, which deserves a more detailed investigation to characterize their possible pathogenicity to humans and animals. Highlights • More than 21,000 sand flies, collected in Northern Italy between 2013 and 2016, were tested. • Toscana, Fermo-like, Corfou, Ponticelli, and two unknown viruses were detected. • The three reassortant Ponticelli I, II, and III viruses were isolated. • The potential pathogenicity of these co-circulating viruses must be investigated. [ABSTRACT FROM AUTHOR]