Artificial membrane

Mosquito strains used in this study included: A. albopictus Gainesville (BEI Resources, NIAID, NIH: MRA-804, contributed by Sandra A. Allan), A. aegypti Liverpool-IB12 (LVP-IB12), A. gambiae G3 (BEI Resources, NIAID, NIH: Eggs, MRA-112, contributed by Mark Q. Benedict), and C. quinquefasciatus JHB (provided by the Centers for Disease Control and Prevention for distribution by BEI Resources, NIAID, NIH: Eggs, NR-43025). These mosquitoes were reared as described [64 (link)] in the insectary, which was maintained at 26.5 °C, with a 12 h dark/12 h light cycle that included 1 h crepuscular periods at the beginning and end of each cycle, and at ~80% relative humidity. An artificial membrane (Hemotek Limited, Blackburn, UK) was used to deliver commercially acquired sheep blood (HemoStat Laboratories, Dixon, CA, USA).

The Ae. aegypti were grown as per Bohbot’s et al. methodology subjected to 12 h daylight and 12 h dark environment cycles while being fed with 10 w/w% sucrose solution in water through a cotton ball (17 (link)). Dried Ae. aegypti eggs were hatched in water and fed with crab food through their larval phase in the same environmental conditions as the mature mosquitoes. Pupae were collected and placed in developmental cages to reach maturity with sucrose feed changed every 48 h. Mature mosquitoes were transferred to experimental cages and split according to the needs of each treatment. Table 1 outlines the population in each cage for the artificial feeding experiments of Ae. aegypti.
For the in vivo experiments on human hand, the mosquitoes were frozen immediately after the experiment allowing monitoring of blood swelling in females thus improving the count. For the artificial membrane (Hemotek) experiments, the mosquitoes were fed ad-libitum for 1 h in the lab environment (20–23 ${}^{\circ }$ C and a relative humidity between 40% and 50%) and immediately placed back in the incubators with a 10 wt.% sucrose solution replaced every 24 h to feed on.