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Fig. 2 | Parasites & Vectors

Fig. 2

From: A novel assay to isolate and quantify third-stage Dirofilaria immitis and Brugia malayi larvae emerging from individual Aedes aegypti

Fig. 2

Assay for eL3. a Infection and assay protocol followed in this study. Mosquitoes are fed on blood containing Dirofilaria immitis microfilariae. Uptake is measured in a small group, and the remaining mosquitoes are maintained until an emergence assay is performed. Immediately following the emergence assay with whole mosquitoes, the Malpighian tubules are dissected and analyzed. The dissected head and carcass are individually placed into an emergence assay to assay L3 that failed to emerge from intact mosquitoes. Additional file 2: Figure S1 shows images of dissected mosquitoes and Malpighian tubules. b, c Mosquitoes are rinsed with 70% ethanol:30% water for wetting, rinsed with water (b) and then placed individually into wells of a 96-well plate (c). d Emerging eL3 from intact mosquitoes or L3 from dissected heads and carcasses at the bottom each well are scored by microscopy. Additional file 3: Movie 2 shows typical movement of eL3. e Larvae (white arrows) in live Malpighian tubules are scored by microscopy. Scale bar: 50 µm

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