Glytube: A Conical Tube and Parafilm M-Based Method as a Simplified Device to Artificially Blood-Feed the Dengue Vector Mosquito, Aedes aegypti

Abstract

Aedes aegypti, the main vector of dengue virus, requires a blood meal to produce eggs. Although live animals are still the main blood source for laboratory colonies, many artificial feeders are available. These feeders are also the best method for experimental oral infection of Ae. aegypti with Dengue viruses. However, most of them are expensive or laborious to construct. Based on principle of Rutledge-type feeder, a conventional conical tube, glycerol and Parafilm-M were used to develop a simple in-house feeder device. The blood feeding efficiency of this apparatus was compared to a live blood source, mice, and no significant differences (p = 0.1189) were observed between artificial-fed (51.3% of engorgement) and mice-fed groups (40.6%). Thus, an easy to assemble and cost-effective artificial feeder, designated “Glytube” was developed in this report. This simple and efficient feeding device can be built with common laboratory materials for research on Ae. aegypti.

A Peptide Signaling System that Rapidly Enforces Paternity in the Aedes aegypti Mosquito

“…Adult mosquitoes were housed with siblings in BugDorm-1 (Bugdorm) or custom cages and provided constant access to 10% sucrose. Adult females were blood-fed on mice for stock maintenance, on human subjects for HP-I mutant generation, and on human subjects or sheep blood delivered via Glytube membrane feeders [31] for egg-laying and host-seeking experiments. Mosquitoes were sexed and sorted under cold anesthesia (4°C).…”

“…For experiments in Figure S1, Figure 2, and Figure 3, females were fed sheep blood in groups of 20–50 using Glytube membrane feeders exactly as described [31]. Glytubes were placed on top of mesh on the mosquito cage, and females were allowed to feed through the mesh for 15 min.…”

“…Groups of 7 to 14 day-old female mosquitoes were fed sheep blood using Glytube membrane feeders [31]. Immediately after blood-feeding, individual mosquitoes were placed in plastic Drosophila vials (25 mm diameter, 95 mm long) containing 5 ml water and a Whatman filter paper (55 mm diameter; GE Healthcare) folded into a cone to act as an oviposition substrate.…”

General visual and contingent thermal cues interact to elicit attraction in female Aedes aegypti mosquitoes

“…For experiments in Figure 1, females were fed sheep blood or saline in groups of 20-50 using Glytube membrane feeders as described [34]. The protein-free saline meal contained 110 mM NaCl, 20 mM NaHCO3, and 1 mM ATP.…”

Implementation of the Artificial Feeders in Hematophagous Arthropod Research Cooperates to the Vertebrate Animal Use Replacement, Reduction and Refinement (3Rs) Principle

“…Based on the pioneering apparatus, a Rutledge feeder [7], many articles have addressed this issue in the previous decades and a large proportion of them developed creative adaptations to promote the construction of different feeders for blood-sucking invertebrate organisms with easily accessible materials [8][9][10][11][12][13][14][15][16][17][18][19][20]. Most of these feeders are composed of a heating element (for blood warming) and a feeding part containing a blood reservoir and an artificial membrane surface simulating a vertebrate skin.…”