Mosquitoes and blood feeding procedure

Ae. aegypti (NIH-Rockefeller strain) mosquitoes were maintained in Percival Intellus I-41VL incubators (Percival Scientific, Perry, IA, USA) and in a Caron 6015 Insect Growth Chamber connected to a Caron CRSY 102 condensate recirculating system (Caron Products and Services, Marietta, OH, USA) at 28°C and 75% relative humidity with a light–dark cycle of 16:8 h. Mosquitoes were reared as previously described (19). Females 3 to 4 d old were allowed to feed on either a bovine blood meal or a bovine blood meal supplemented with 3 mM allopurinol (the maximal concentration soluble in blood) for 15 min via an artificial blood feeder connected to a 37°C water bath. Only females that fed to repletion were used for experiments. All females were maintained on 3% sucrose before and after blood feeding. Experiments were replicated at least 3 times with 3 separate cohorts of mosquitoes.