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This holometabolous insect has midgut digestive enzymes that are trypsin\u2010 and chymotrypsin\u2010like proteins. Protease inhibitors are able to bind to proteolytic enzymes and promote a blockage in digestion and nutrition, leading to death. Thus, we investigated the effect of trypsin inhibitor of <jats:italic>Leucaena leucocephala<\/jats:italic> (<jats:styled-content style=\"fixed-case\">LTI<\/jats:styled-content>) seeds on egg hatching, larval development and digestive midgut proteases.<\/jats:p><\/jats:sec><jats:sec><jats:title>RESULTS<\/jats:title><jats:p><jats:styled-content style=\"fixed-case\">LTI<\/jats:styled-content> was obtained by trichloroacetic acid precipitation followed by a single chromatography step on anhydrous trypsin sepharose. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate showed a single protein band with a molecular mass close to 20 <jats:styled-content style=\"fixed-case\">kDa<\/jats:styled-content>. After exposure of <jats:italic>Ae. aegypti<\/jats:italic> eggs to <jats:styled-content style=\"fixed-case\">LTI<\/jats:styled-content> (0.3\u2009mg <jats:styled-content style=\"fixed-case\">mL<\/jats:styled-content><jats:sup>\u22121<\/jats:sup>), egg hatching was reduced (50%). <jats:styled-content style=\"fixed-case\">LTI<\/jats:styled-content> did not show acute toxicity on newly hatched larvae incubated under the same conditions, but after 10 days of exposure a high mortality rate (86%) was observed and the surviving larvae had a 25% delay in development. <jats:styled-content style=\"fixed-case\">LTI<\/jats:styled-content> was able to inhibit <jats:italic>in vitro<\/jats:italic> the midgut enzymatic activity (70%), and when larvae were incubated with <jats:styled-content style=\"fixed-case\">LTI<\/jats:styled-content> solution we observed an inhibition of 56%.<\/jats:p><\/jats:sec><jats:sec><jats:title>CONCLUSIONS<\/jats:title><jats:p><jats:styled-content style=\"fixed-case\">LTI<\/jats:styled-content> is a promising new tool to control critical points of <jats:italic>Ae. aegypti<\/jats:italic> development. \u00a9 2016 Society of Chemical Industry<\/jats:p><\/jats:sec>","DOI":"10.1002\/ps.4284","type":"journal-article","created":{"date-parts":[[2016,4,4]],"date-time":"2016-04-04T07:33:44Z","timestamp":1459755224000},"page":"181-187","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Trypsin inhibitor from <i>Leucaena leucocephala<\/i> seeds delays and disrupts the development of <i>Aedes aegypti,<\/i> a multiple\u2010disease vector"],"prefix":"10.1002","volume":"73","author":[{"given":"Luiz CP","family":"Almeida Filho","sequence":"first","affiliation":[{"name":"Biochemistry and Molecular Biology Department Federal University of Cear\u00e1  Fortaleza Cear\u00e1 Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Terezinha M","family":"de Souza","sequence":"additional","affiliation":[{"name":"Biochemistry and Molecular Biology Department Federal University of Cear\u00e1  Fortaleza Cear\u00e1 Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pedro MS","family":"Tabosa","sequence":"additional","affiliation":[{"name":"Biology Department Federal University of Cear\u00e1  Fortaleza Cear\u00e1 Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nayana G","family":"Soares","sequence":"additional","affiliation":[{"name":"Biology Department Federal University of Cear\u00e1  Fortaleza Cear\u00e1 Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lady CB","family":"Rocha\u2010Bezerra","sequence":"additional","affiliation":[{"name":"Biochemistry and Molecular Biology Department Federal University of Cear\u00e1  Fortaleza Cear\u00e1 Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ilka M","family":"Vasconcelos","sequence":"additional","affiliation":[{"name":"Biochemistry and Molecular Biology Department Federal University of Cear\u00e1  Fortaleza Cear\u00e1 Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ana FU","family":"Carvalho","sequence":"additional","affiliation":[{"name":"Biology Department Federal University of Cear\u00e1  Fortaleza Cear\u00e1 Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2016,5,19]]},"reference":[{"key":"e_1_2_5_2_1","first-page":"299","article-title":"Epidemiology of dengue: past, present and future prospects","volume":"5","author":"Murray NEA","year":"2013","journal-title":"Clin Epidemiol"},{"key":"e_1_2_5_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.apjtm.2015.06.009"},{"key":"e_1_2_5_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jfma.2014.12.002"},{"key":"e_1_2_5_5_1","doi-asserted-by":"publisher","DOI":"10.1186\/1756-3305-6-351"},{"key":"e_1_2_5_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ibmb.2012.10.007"},{"key":"e_1_2_5_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.medmal.2014.04.008"},{"key":"e_1_2_5_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/uog.15831"},{"key":"e_1_2_5_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ecolmodel.2015.07.007"},{"key":"e_1_2_5_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.actatropica.2015.08.013"},{"key":"e_1_2_5_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pestbp.2014.09.014"},{"key":"e_1_2_5_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.procbio.2015.05.022"},{"key":"e_1_2_5_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.aspen.2014.05.003"},{"key":"e_1_2_5_14_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00436-013-3640-8"},{"key":"e_1_2_5_15_1","first-page":"227","article-title":"Larvicidal activity of affinin and its derived amides from Heliopsis longipes A","volume":"18","author":"Hern\u00e1ndez\u2010Morales A","year":"2015","journal-title":"Gray Blake against Anopheles albimanus and Aedes aegypti. 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