{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,12]],"date-time":"2026-09-12T12:48:57Z","timestamp":1789217337045,"version":"build-2803163510"},"reference-count":41,"publisher":"American Society for Microbiology","issue":"5","license":[{"start":{"date-parts":[[2023,10,17]],"date-time":"2023-10-17T00:00:00Z","timestamp":1697500800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2023,10,17]],"date-time":"2023-10-17T00:00:00Z","timestamp":1697500800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100004040","name":"KU Leuven","doi-asserted-by":"crossref","award":["C22\/18\/007, STG\/19\/008"],"award-info":[{"award-number":["C22\/18\/007, STG\/19\/008"]}],"id":[{"id":"10.13039\/501100004040","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"published-print":{"date-parts":[[2023,10,17]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    <jats:italic toggle=\"yes\">Aedes aegypti<\/jats:italic>\n                    mosquitoes can transmit several arboviruses, including chikungunya virus (CHIKV), dengue virus (DENV), and Zika virus (ZIKV). When blood-feeding on a virus-infected human, the mosquito ingests the virus into the midgut (stomach), where it replicates and must overcome the midgut barrier to disseminate to other organs and ultimately be transmitted via the saliva. Current tools to study mosquito-borne viruses (MBVs) include 2D-cell culture systems and\n                    <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                    mosquito infection models, which offer great advantages, yet have some limitations. Here, we describe a long-term\n                    <jats:italic toggle=\"yes\">ex vivo<\/jats:italic>\n                    culture of\n                    <jats:italic toggle=\"yes\">Ae. aegypti<\/jats:italic>\n                    guts. Cultured guts were metabolically active for 7 d in a 96-well plate at 28\u00b0C and were permissive to ZIKV, DENV, Ross River virus, and CHIKV.\n                    <jats:italic toggle=\"yes\">Ex vivo<\/jats:italic>\n                    guts from\n                    <jats:italic toggle=\"yes\">Culex pipiens<\/jats:italic>\n                    mosquitoes were found to be permissive to Usutu virus. Immunofluorescence staining confirmed viral protein synthesis in CHIKV-infected guts of\n                    <jats:italic toggle=\"yes\">Ae. aegypti<\/jats:italic>\n                    . Furthermore, fluorescence microscopy revealed replication and spread of a reporter DENV in specific regions of the midgut. In addition, two known antiviral molecules, \u03b2-\n                    <jats:sc>d<\/jats:sc>\n                    -N\n                    <jats:sup>4<\/jats:sup>\n                    -hydroxycytidine and 7-deaza-2\u2032-C-methyladenosine, were able to inhibit CHIKV and ZIKV replication, respectively, in the\n                    <jats:italic toggle=\"yes\">ex vivo<\/jats:italic>\n                    model. Together, our results show that\n                    <jats:italic toggle=\"yes\">ex vivo<\/jats:italic>\n                    guts can be efficiently infected with mosquito-borne alpha- and flaviviruses and employed to evaluate antiviral drugs. Furthermore, the setup can be extended to other mosquito species.\n                    <jats:italic toggle=\"yes\">Ex vivo<\/jats:italic>\n                    gut cultures could thus be a new model to study MBVs, offering the advantage of reduced biosafety measures compared to infecting living mosquitoes.\n                  <\/jats:p>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>\n                      Mosquito-borne viruses (MBVs) are a significant global health threat since they can cause severe diseases in humans, such as hemorrhagic fever, encephalitis, and chronic arthritis. MBVs rely on the mosquito vector to infect new hosts and perpetuate virus transmission. No therapeutics are currently available. The study of arbovirus infection in the mosquito vector can greatly contribute to elucidating strategies for controlling arbovirus transmission. This work investigated the infection of guts from\n                      <jats:italic toggle=\"yes\">Aedes aegypti<\/jats:italic>\n                      mosquitoes in an\n                      <jats:italic toggle=\"yes\">ex vivo<\/jats:italic>\n                      platform. We found several MBVs capable of replicating in the gut tissue, including viruses of major health importance, such as dengue, chikungunya, and Zika viruses. In addition, antiviral compounds reduced arbovirus infection in the cultured gut tissue. Overall, the gut model emerges as a useful tool for diverse applications such as studying tissue-specific responses to virus infection and screening potential anti-arboviral molecules.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.05195-22","type":"journal-article","created":{"date-parts":[[2023,8,4]],"date-time":"2023-08-04T09:00:18Z","timestamp":1691139618000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":9,"title":["<i>Ex vivo<\/i>\n                    gut cultures of\n                    <i>Aedes aegypti<\/i>\n                    are efficiently infected by mosquito-borne alpha- and flaviviruses"],"prefix":"10.1128","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2099-9152","authenticated-orcid":false,"given":"Ana Lucia","family":"Rosales Rosas","sequence":"first","affiliation":[{"name":"KU Leuven Department of Microbiology, Immunology and 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