Rearing Black Flies (Diptera: Simuliidae) in the Laboratory: Mass-Scale in Vitro Membrane Feeding and its Application to Collection of Saliva and to Parasitological and Repellent Studies1

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Abstract
A highly efficient, mass-scale in vitro membrane feeding system for black flies is described that has proven effective for 5 Simulium species exhibiting a wide range of blood-feeding preferences. Factors affecting feeding rate, such as type of membrane or blood used, addition of ATP, age of flies, and temperature differential, were examined for the species tested. Nearctic black flies could be stored at a cool temperature (ca. 7 °C) for up to 20 days with no significant decrease in feeding rates, while cool storage was detrimental to S. damnosum s.l., a tropical species. Saliva was collected from females as they fed on various nonblood sources such as tissue culture media, saline solutions, or distilled water. Repellent activity was evaluated for 6 commercial formulations of deet by using a “free-choice” system and counting actual bites through the membrane. Percent reduction of biting activity increased as the percentage of deet in the repellent increased. Attempts to infect simuliid females per os with 3 vertebrate pathogens—Onchocerca lienalis, Leucocytozoon smithi, and vesicular stomatitis virus (New Jersey serotype)—resulted in successful recovery of infective forms from the flies.