Successful forced feeding of females of Phlebotomus papatasi, P. perniciosus, P. argentipes, Lutzomyia longipalpis and L. spinicrassa is reported using the method of A.N. Alexeiev [see Medical Parasitology, Moscow, 34: 467-471 (1965)], but obviating high mortality from anaesthetics by handling the active flies by the wings with fine forceps. In preliminary experiments, P. papatasi which had been initially infected by feeding via chick skin membrane on a defibrinated rabbit blood-Leishmania minor (WR309) suspension became increasingly unable to imbibe from the microcapillary of Alexeiev's method due to blockage of the proventriculus, oesophagus and pharyngeal armature regions as the infection progressed. Then, even though the flies held in the force-feeding position repeatedly pump with the cibarium, little or no blood may move but Leishmania flagellates may nevertheless appear in the medium in the capillary tube. This first occurred 19 days after membrane-fed infection and 14 days after infection of flies subsequently force-fed. Flagellates were once seen entering the capillary medium suggesting their own movement rather than expulsion. Prior to this, sandflies capable of re-feeding 11 to 14 days after infection expelled 1-50 motile flagellates in anal droplets while engorging. The technique has been demonstrated to facilitate a novel approach to studies on the mechanism and mode of transmission, on the infectivity and feeding frequency of wild-caught sandflies and for the harvesting of expelled parasites for biochemical characterisation.
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