Detection of Multiple Blood Feeding in Aedes aegypti (Diptera: Culicidae) During a Single Gonotrophic Cycle Using a Histologic Technique

We evaluated a histologic technique for its usefulness in detecting multiple blood feeding by Aedes aegypti (L.) in a single gonotrophic cycle. To standardize the procedure, we carried out a laboratory study in which 166 mosquitoes imbibed two blood meals at known intervals. Eighty percent (78/98) of the multiple meals were detected when the interval between meals was from 1 to < to = 24 h and the time from the second meal to fixation ranged from 0 to 12 hr. At intervals outside this range, only 34% (23/68) of the multiple meals were detected. Overall, 61% (101/166) of the double meals were detected. Examination of 96 engorged Ae. aegypti collected by aspiration from inside houses in San Juan, Puerto Rico, indicated that 50% had imbibed multiple meals. Most wild-caught mosquitoes took their last meal the day before capture, and most multiple feeders fed twice on consecutive days. A dark line of digested blood, or heme, around the first meal and a physical separation between meals were the most useful histologic parameters for detecting multiple feeding in wild Ae. aegypti. An association of multiple feeding with advanced stages of oocyte development suggests that, at the time of collection, most Ae. aegypti from the study site had fed twice in each gonotrophic cycle. We conclude that, although it is labor intensive, histologic examination is an appropriate technique for a longitudinal, community-wide survey of multiple feeding by Ae. aegypti.