To assess differences in host-location behaviour between the mosquito strains, including the time to locate a human host, “free-flight” assays were used. Pools of 10, 3–4 d old female mosquitoes, deprived of sugar solution for 24 h were released into a white 18 m3 room (2 x 2.8 x 3.2 m) maintained at 26 ± 1°C and 70% RH. Mosquitoes were allowed to acclimatise and rest for 10 min before a human observer entered and sat in the middle of the room (QIMR Berghofer Human Research Ethics approval P2273). Only their lower legs were exposed, ensuring accurate recording of successful landing and feeding events. Each observation was made over 10 min, although most mosquitoes had landed and probed within 3 min. Time to locate the host was recorded for each mosquito and all probing mosquitoes were removed from further observation by aspiration and placed in a holding container. After 10 min, all remaining mosquitoes were captured. Three batches of 10 mosquitoes from each strain were tested. Each test was conducted between 1700 and 1730 h.

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