Artificial Infection of Sand Flies with VSV

Three-day-old female sand flies and private of sugar solution food for a period of 24h were used in this study. The insects were fed with heparinized blood through chick skin membrane attached in an artificial feeder (Hemotek). The artificial infection was performed with blood containing VSV or a control without virus at 37°C. Fully engorged females were selected and harvested individually at 2, 4 and 6 dpf. Insects were anesthetized with carbon dioxide and directly ground in TRIzol (Invitrogen) using glass beads as previously described [23 (link)].

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Ferreira F.V., Aguiar E.R., Olmo R.P., de Oliveira K.P., Silva E.G., Sant'Anna M.R., Gontijo N.D., Kroon E.G., Imler J.L, & Marques J.T. (2018). The small non-coding RNA response to virus infection in the Leishmania vector Lutzomyia longipalpis. PLoS Neglected Tropical Diseases, 12(6), e0006569.

Publication 2018

artificial feeder TRIzol

Blood Carbon dioxide Females Food Infection Insects Membrane Sand flies Skin Sugar Trizol Virus

Corresponding Organization :

Other organizations : Universidade Federal de Minas Gerais, Modèles Insectes de l'Immunité Innée

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Variable analysis

independent variables

Infection with VSV or control (without virus)

dependent variables

Harvesting of insects at 2, 4 and 6 days post-feeding (dpf)

control variables

Three-day-old female sand flies
Sugar solution food for a period of 24h
Heparinized blood through chick skin membrane attached in an artificial feeder (Hemotek)
Incubation at 37°C
Anesthetization with carbon dioxide
Direct grinding in TRIzol (Invitrogen) using glass beads

controls

Positive control: Blood containing VSV
Negative control: Blood without virus

Annotations

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