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Transmission to a vertebrate host requires that ingested microfilariae develop into infective third-stage larvae capable of emerging from the mosquito proboscis onto the skin of the host during blood-feeding. Determining the number of microfilariae that successfully develop to infective third-stage larvae in the mosquito host is key to understanding parasite transmission potential and to developing new strategies to block these worms in their vector.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Methods<\/jats:title>\n                <jats:p>We developed a novel method to efficiently assess the number of infective third-stage filarial larvae that emerge from experimentally infected mosquitoes. Following infection, individual mosquitoes were placed in wells of a multi-well culture plate and warmed to 37\u00a0\u00b0C to stimulate parasite emergence. <jats:italic>Aedes aegypti<\/jats:italic> infected with <jats:italic>Dirofilaria immitis<\/jats:italic> were used to determine infection conditions and assay timing. The assay was also tested with <jats:italic>Brugia malayi<\/jats:italic>-infected <jats:italic>Ae. aegypti<\/jats:italic>.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>Approximately 30% of <jats:italic>Ae. aegypti<\/jats:italic> infected with <jats:italic>D. immitis<\/jats:italic> and 50% of those infected with <jats:italic>B. malayi<\/jats:italic> produced emerging third-stage larvae. Once <jats:italic>D. immitis<\/jats:italic> third-stage larvae emerged at 13\u00a0days post infection, the proportion of mosquitoes producing them and the number produced per mosquito remained stable until at least day 21. The prevalence and intensity of emerging third-stage <jats:italic>B. malayi<\/jats:italic> were similar on days 12\u201314 post infection. Increased uptake of <jats:italic>D. immitis<\/jats:italic> microfilariae increased the fitness cost to the mosquito but did not increase the number of emerging third-stage larvae.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Conclusions<\/jats:title>\n                <jats:p>We provide a new assay with an associated set of infection conditions that will facilitate assessment of the filarial transmission potential of mosquito vectors and promote preparation of uniformly infectious third-stage larvae for functional assays. The ability to quantify infection outcome will facilitate analyses of molecular interactions between vectors and filariae, ultimately allowing for the establishment of novel methods to block disease transmission.<\/jats:p>\n                \n              <\/jats:sec>","DOI":"10.1186\/s13071-020-04529-w","type":"journal-article","created":{"date-parts":[[2021,1,7]],"date-time":"2021-01-07T12:03:47Z","timestamp":1610021027000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["A novel assay to isolate and quantify third-stage Dirofilaria immitis and Brugia malayi larvae emerging from individual Aedes aegypti"],"prefix":"10.1186","volume":"14","author":[{"given":"Abigail R.","family":"McCrea","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elizabeth B.","family":"Edgerton","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Genevieve T.","family":"Oliver","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fiona M.","family":"O\u2019Neill","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas J.","family":"Nolan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"James B.","family":"Lok","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3766-2097","authenticated-orcid":false,"given":"Michael","family":"Povelones","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,1,7]]},"reference":[{"issue":"9747","key":"4529_CR1","doi-asserted-by":"publisher","first-page":"1175","DOI":"10.1016\/S0140-6736(10)60586-7","volume":"376","author":"MJ Taylor","year":"2010","unstructured":"Taylor MJ, Hoerauf A, Bockarie M. 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