{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T09:55:33Z","timestamp":1790502933009,"version":"4.1.0"},"reference-count":50,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2019,4,9]],"date-time":"2019-04-09T00:00:00Z","timestamp":1554768000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100007397","name":"Univerzita Karlova v Praze","doi-asserted-by":"publisher","award":["UNCE\/SCI\/012"],"award-info":[{"award-number":["UNCE\/SCI\/012"]}],"id":[{"id":"10.13039\/100007397","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Centre for Research of pathogenicity and virulence of parasites","award":["16_019\/0000759"],"award-info":[{"award-number":["16_019\/0000759"]}]},{"DOI":"10.13039\/501100000921","name":"European Cooperation in Science and Technology","doi-asserted-by":"publisher","award":["TD1303, EurNegVec"],"award-info":[{"award-number":["TD1303, EurNegVec"]}],"id":[{"id":"10.13039\/501100000921","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100010666","name":"H2020 Research Infrastructures","doi-asserted-by":"publisher","award":["731060, INFRAVEC2"],"award-info":[{"award-number":["731060, INFRAVEC2"]}],"id":[{"id":"10.13039\/100010666","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100010666","name":"H2020 Research Infrastructures","doi-asserted-by":"publisher","award":["653316, European Virus Archive goes Global (EVAg)"],"award-info":[{"award-number":["653316, European Virus Archive goes Global (EVAg)"]}],"id":[{"id":"10.13039\/100010666","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Background: Massilia virus (MASV) is a phlebovirus isolated from Phlebotomus perniciosus in various regions of southwestern Europe. It is closely related to human pathogens such as Toscana virus and sandfly fever Naples virus. The natural cycle of phleboviruses is poorly understood. Indeed, experimental studies demonstrate that transovarial and sexual transmission are not efficient enough for the maintenance of the virus in nature and to date there is no convincing evidence that a species of vertebrates is the reservoir of the virus. Here, we studied various transmission routes of MASV taking advantage of experimental colonies representing different species of sand flies. Methodology\/Principal findings: In P. perniciosus, four sources of infection were compared: (i) Virus-seeded larval food to the first instar larvae (L1), or (ii) to the fourth instar larvae (L4), (iii) virus-seeded blood meal to adult females, and (iv) virus-seeded sugar meal to adults of both sexes. From 875 adults emerged from infected L1 and L4, only three were positive. In females infected by bloodmeal the infection rate was high before defecation, then it decreased drastically; MASV RNA was detected in only 5 out of 27 post-defecation. Surprisingly, the most efficient route of infection was observed after intake of virus-seeded sugar meal: 72% of females (79\/110) and 52% of males (51\/99) were found to be MASV RNA-positive. In addition, MASV-infected sandflies regurgitated virus particules into the sugar drop and MASV RNA was detectable in this drop for at least 24 h after regurgitation. MASV RNA was detected in about one third of the P. perniciosus exposed to this sugar drop contaminated by regurgitation. Sugar meal infection was also tested with six other species of sand flies. In males, there were no significant differences in infection rates when compared to P. perniciosus. In females, most species tested showed high infection rate at the beginning but then significant gradual decrease in infection rate during the experiment. Conclusions\/Significance: We present the first description of arboviral infection of a dipteran vector using sugar meal. In all seven sand fly species tested, MASV was detected for two weeks post-infection. Our results showed that MASV can be transmitted between P. perniciosus either through co-feeding or via an infected sugar source such as plant sap. These newly described routes of horizontal transmission may play an important role in the circulation of phleboviruses in nature.<\/jats:p>","DOI":"10.3390\/v11040332","type":"journal-article","created":{"date-parts":[[2019,4,9]],"date-time":"2019-04-09T05:58:07Z","timestamp":1554789487000},"page":"332","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Experimental Infection of Sand Flies by Massilia Virus and Viral Transmission by Co-Feeding on Sugar Meal"],"prefix":"10.3390","volume":"11","author":[{"given":"Magdalena","family":"Jancarova","sequence":"first","affiliation":[{"name":"Department of Parasitology, Faculty of Science, Charles University, 128 44 Prague 2, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laurence","family":"Bichaud","sequence":"additional","affiliation":[{"name":"Department of Parasitology, Faculty of Science, Charles University, 128 44 Prague 2, Czech Republic"},{"name":"Unit\u00e9 des Virus Emergents (UVE: Aix Marseille Univ, IRD 190, INSERM 1207, IHU M\u00e9diterran\u00e9e Infection), 13005 Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jana","family":"Hlavacova","sequence":"additional","affiliation":[{"name":"Department of Parasitology, Faculty of Science, Charles University, 128 44 Prague 2, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7102-3654","authenticated-orcid":false,"given":"Stephane","family":"Priet","sequence":"additional","affiliation":[{"name":"Unit\u00e9 des Virus Emergents (UVE: Aix Marseille Univ, IRD 190, INSERM 1207, IHU M\u00e9diterran\u00e9e Infection), 13005 Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nazli","family":"Ayhan","sequence":"additional","affiliation":[{"name":"Unit\u00e9 des Virus Emergents (UVE: Aix Marseille Univ, IRD 190, INSERM 1207, IHU M\u00e9diterran\u00e9e Infection), 13005 Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tatiana","family":"Spitzova","sequence":"additional","affiliation":[{"name":"Department of Parasitology, Faculty of Science, Charles University, 128 44 Prague 2, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Petr","family":"Volf","sequence":"additional","affiliation":[{"name":"Department of Parasitology, Faculty of Science, Charles University, 128 44 Prague 2, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7675-8251","authenticated-orcid":false,"given":"Remi N.","family":"Charrel","sequence":"additional","affiliation":[{"name":"Unit\u00e9 des Virus Emergents (UVE: Aix Marseille Univ, IRD 190, INSERM 1207, IHU M\u00e9diterran\u00e9e Infection), 13005 Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2019,4,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1016\/S0738-081X(99)00046-2","article-title":"The biology and control of phlebotomine sand flies","volume":"17","year":"1999","journal-title":"Clin. Dermatol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1111\/j.1365-2915.2012.01034.x","article-title":"Phlebotomine sandflies and the spreading of leishmaniases and other diseases of public health concern","volume":"27","author":"Maroli","year":"2013","journal-title":"Med. Vet. Entomol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1016\/j.antiviral.2013.07.005","article-title":"Sandfly-borne phleboviruses of Eurasia and Africa: Epidemiology, genetic diversity, geographic range, control measures","volume":"100","author":"Alkan","year":"2013","journal-title":"Antivir. Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"927","DOI":"10.1007\/s00705-018-04127-3","article-title":"Taxonomy of the order Bunyavirales: Second update 2018","volume":"164","author":"Maes","year":"2019","journal-title":"Arch. Virol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1007\/s00436-012-2910-1","article-title":"Tick-borne viruses in Europe","volume":"111","author":"Hubalek","year":"2012","journal-title":"Parasitol. Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"4373","DOI":"10.3390\/v6114373","article-title":"Bunyavirus-vector interactions","volume":"6","author":"Horne","year":"2014","journal-title":"Viruses"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1657","DOI":"10.3201\/eid1111.050869","article-title":"Emergence of Toscana virus in Europe","volume":"11","author":"Charrel","year":"2005","journal-title":"Emerg. Infect. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"135","DOI":"10.5501\/wjv.v1.i5.135","article-title":"Emergence of Toscana virus in the mediterranean area","volume":"1","author":"Charrel","year":"2012","journal-title":"World J. Virol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1186\/s13071-015-0826-1","article-title":"Toscana virus isolated from sandflies, Morocco","volume":"8","author":"Ajaoud","year":"2015","journal-title":"Parasit. Vectors"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1299","DOI":"10.3201\/eid1708.060345","article-title":"Toscana virus RNA in Sergentomyia minuta files","volume":"12","author":"Charrel","year":"2006","journal-title":"Emerg. Infect. Dis."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s13071-014-0575-6","article-title":"Phlebovirus and Leishmania detection in sandflies from eastern Thrace and northern Cyprus","volume":"7","author":"Ergunay","year":"2014","journal-title":"Parasit. Vectors"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"336","DOI":"10.3389\/fpubh.2017.00336","article-title":"Cocirculation of Two Lineages of Toscana Virus in Croatia","volume":"12","author":"Ayhan","year":"2017","journal-title":"Front. Public Health"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"86","DOI":"10.4103\/0972-9062.134785","article-title":"Phlebotomus sergenti a common vector of Leishmania tropica and Toscana virus in Morocco","volume":"51","author":"Ajaoud","year":"2014","journal-title":"J. Vector Borne Dis."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"519","DOI":"10.1089\/vbz.2008.0131","article-title":"Massilia virus, a novel Phlebovirus (Bunyaviridae) isolated from sandflies in the Mediterranean","volume":"9","author":"Charrel","year":"2009","journal-title":"Vector Borne Zoonotic Dis."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1111\/j.1749-6632.1975.tb35093.x","article-title":"Transovarial transmission of viruses by phlebotomine sandflies","volume":"266","author":"Tesh","year":"1975","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"189","DOI":"10.4269\/ajtmh.1987.36.189","article-title":"Maintenance of Toscana virus in Phlebotomus perniciosus by vertical transmission","volume":"36","author":"Tesh","year":"1987","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"574","DOI":"10.4269\/ajtmh.1992.47.574","article-title":"Simulation of arbovirus overwintering: Survival of Toscana virus (Bunyaviridae: Phlebovirus) in its natural sand fly vector Phlebotomus perniciosus","volume":"47","author":"Tesh","year":"1992","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1111\/j.1365-2915.1993.tb00689.x","article-title":"Vertical transmission of Toscana virus in the sandfly, Phlebotomus perniciosus, via the second gonotrophic cycle","volume":"7","author":"Maroli","year":"1993","journal-title":"Med. Vet. Entomol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"802","DOI":"10.1007\/BF01923553","article-title":"Non-viraemic transmission of tick-borne encephalitis virus: A mechanism for arbovirus survival in nature","volume":"49","author":"Labuda","year":"1993","journal-title":"Experientia"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"8871","DOI":"10.1073\/pnas.0503835102","article-title":"Nonviremic transmission of West Nile virus","volume":"102","author":"Higgs","year":"2005","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"485","DOI":"10.1126\/science.287.5452.485","article-title":"Transmission of vesicular stomatitis virus from infected to noninfected black flies co-feeding on nonviremic deer mice","volume":"287","author":"Mead","year":"2000","journal-title":"Science"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1111\/j.1948-7134.2011.00106.x","article-title":"Establishment and maintenance of sand fly colonies","volume":"36","author":"Volf","year":"2011","journal-title":"J. Vector Ecol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2816","DOI":"10.1099\/jgv.0.000592","article-title":"Neutralization-based seroprevalence of Toscana virus and sandfly fever Sicilian virus in dogs and cats from Portugal","volume":"97","author":"Alwassouf","year":"2016","journal-title":"J. Gen. Virol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"690","DOI":"10.4269\/ajtmh.14-0322","article-title":"Potential animal reservoirs of Toscana virus and coinfections with Leishmania infantum in Turkey","volume":"92","author":"Dincer","year":"2015","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"370","DOI":"10.1099\/vir.0.025940-0","article-title":"Horizontal transmission of deformed wing virus: Pathological consequences in adult bees (Apis mellifera) depend on the transmission route","volume":"92","author":"Gisder","year":"2011","journal-title":"J. Gen. Virol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Ferreira, \u00c1.G., Naylor, H., Esteves, S.S., Pais, I.S., Martins, N.E., and Teixeira, L. (2014). The Toll-dorsal pathway is required for resistance to viral oral infection in Drosophila. PLoS Pathog., 10.","DOI":"10.1371\/journal.ppat.1004507"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1490","DOI":"10.1099\/vir.0.000068","article-title":"Infectivity of Drosophila C virus following oral delivery in Drosophila larvae","volume":"96","author":"Stevanovic","year":"2015","journal-title":"J. Gen. Virol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1146\/annurev.en.28.010183.001305","article-title":"Intrinsic factors affecting vector competence of mosquitoes for arboviruses","volume":"28","author":"Hardy","year":"1983","journal-title":"Annu. Rev. Entomol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"3741","DOI":"10.3390\/v7072795","article-title":"Tissue barriers to arbovirus infection in mosquitoes","volume":"7","author":"Franz","year":"2015","journal-title":"Viruses"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"185","DOI":"10.4103\/0972-9062.166248","article-title":"Vector competence of two Indian populations of Culex quinquefasciatus (Diptera: Culicidae) mosquitoes to three West Nile virus strains","volume":"52","author":"Sudeep","year":"2015","journal-title":"J. Vector Borne Dis."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"151","DOI":"10.4269\/ajtmh.2000.62.151","article-title":"Vertical transmission of the yellow fever virus by Aedes aegypti (Diptera, Culicidae): Dynamics of infection in F1 adult progeny of orally infected females","volume":"62","author":"Diallo","year":"2000","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_32","first-page":"131","article-title":"Susceptibility and transovarial transmission of dengue virus in Aedes aegypti: A preliminary study of morphological variations","volume":"34","author":"Wasinpiyamongkol","year":"2003","journal-title":"Southeast Asian J. Trop. Med. Public Health"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"809","DOI":"10.1590\/S0074-02762004000800005","article-title":"Dengue virus detection by using reverse transcription-polymerase chain reaction in saliva and progeny of experimentally infected Aedes albopictus from Brazil","volume":"99","author":"Castro","year":"2004","journal-title":"Mem\u00f3rias Inst. Oswaldo Cruz"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1031","DOI":"10.1603\/ME11043","article-title":"Evidence of efficient transovarial transmission of Culex flavivirus by Culex pipiens (Diptera: Culicidae)","volume":"48","author":"Saiyasombat","year":"2011","journal-title":"J. Med. Entomol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"73","DOI":"10.14411\/fp.2002.014","article-title":"Bacterial colonisation in the gut of Phlebotomus duboscqi (Diptera: Psychodidae): Transtadial passage and the role of female diet","volume":"49","author":"Volf","year":"2002","journal-title":"Folia Parasitol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1186\/s13071-015-0635-6","article-title":"Malpighian tubules are important determinants of Pseudomonas transstadial transmission and longtime persistence in Anopheles stephensi","volume":"8","author":"Chavshin","year":"2015","journal-title":"Parasit. Vector"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"593","DOI":"10.1146\/annurev-ento-112408-085450","article-title":"Regulation of midgut growth, development, and metamorphosis","volume":"55","author":"Hakim","year":"2010","journal-title":"Ann. Rev. Entomol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"506","DOI":"10.1016\/j.parint.2014.01.004","article-title":"Aedes aegypti midgut remodeling during metamorphosis","volume":"63","author":"Fernandes","year":"2014","journal-title":"Parasitol. Int."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1007\/s13199-013-0231-5","article-title":"Escherichia coli expressing a green fluorescent protein (GFP) in Anopheles stephensi: A preliminary model for paratransgenesis","volume":"60","author":"Chavshin","year":"2013","journal-title":"Symbiosis"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1146\/annurev.en.35.010190.001145","article-title":"Evolution of digestive systems of insects","volume":"35","author":"Terra","year":"1990","journal-title":"Ann. Rev. Entomol."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Peters, W. (1992). Peritrophic Membranes, Springer Press.","DOI":"10.1007\/978-3-642-84414-0"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"525","DOI":"10.1146\/annurev.ento.42.1.525","article-title":"Peritrophic matrix structure and function","volume":"42","author":"Lehane","year":"1997","journal-title":"Ann. Rev. Entomol."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Pruzinova, K., Sadlova, J., Seblova, V., Homola, M., Votypka, J., and Volf, P. (2015). Comparison of bloodmeal digestion and the peritrophic matrix in four sand fly species differing in susceptibility to Leishmania donovani. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0128203"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Xi, Z., Ramirez, J.L., and Dimopoulos, G. (2008). The Aedes aegypti toll pathway controls dengue virus infection. PLoS Pathog., 4.","DOI":"10.1371\/journal.ppat.1000098"},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Ramirez, J.L., Souza-Neto, J., Cosme, R.T., Rovira, J., Ortiz, A., Pascale, J.M., and Dimopoulos, G. (2012). Reciprocal tripartite interactions between the Aedes aegypti midgut microbiota, innate immune system and dengue virus influences vector competence. PLoS Negl. Trop. Dis., 6.","DOI":"10.1371\/journal.pntd.0001561"},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Ramirez, J.L., Short, S.M., Bahia, A.C., Saraiva, R.G., Dong, Y., Kang, S., Tripathi, A., Mlambo, G., and Dimopoulos, G. (2014). Chromobacterium Csp_P reduces malaria and dengue infection in vector mosquitoes and has entomopathogenic and in vitro anti-pathogen activities. PLoS Pathog., 10.","DOI":"10.1371\/journal.ppat.1004398"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1046\/j.1365-2915.1998.00063.x","article-title":"Sugar feeding and fluid destination control in the phlebotomine sandfly Lutzomyia longipalpis (Diptera: Psychodidae)","volume":"12","author":"Tang","year":"1998","journal-title":"Med. Vet. Entomol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"845","DOI":"10.1093\/jmedent\/44.5.845","article-title":"Expectoration of flaviviruses during sugar feeding by mosquitoes (Diptera: Culicidae)","volume":"44","author":"Johnson","year":"2007","journal-title":"J. Med. Entomol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1111\/j.1365-2915.1987.tb00373.x","article-title":"Honeydew sugars in wild-caught Phlebotomus ariasi detected by high performance liquid chromatography (HPLC) and gas chromatography (GC)","volume":"1","author":"Moore","year":"1987","journal-title":"Med. Vet. Entomol."},{"key":"ref_50","first-page":"431","article-title":"Sugars in sandflies","volume":"33","author":"Molyneux","year":"1991","journal-title":"Parassitologia"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/11\/4\/332\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T12:43:51Z","timestamp":1760186631000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/11\/4\/332"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,4,9]]},"references-count":50,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2019,4]]}},"alternative-id":["v11040332"],"URL":"https:\/\/doi.org\/10.3390\/v11040332","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,4,9]]}}}