Histologic detection of multiple blood meals in Phlebotomus duboscqi (Diptera: Psychodidae)

A histologic technique was used to detect multiple hamster blood meals taken by Phlebotomus duboscqi Neveu-Lemaire during a 5-d period. Forty-eight flies were fed two or three blood meals separated by 48, 72, or 120 h and sampled immediately; multiple meals were detected in 27 flies (56%). Double meals separated by 72 h within a single gonotrophic cycle were documented in 11/19 (58%) flies; double meals separated by 120 h were detected in only 4/17 (24%) flies. Triple blood meals taken at 0, 72, and 120 h were detected in 5/12 (42%) flies; all of these flies contained the second and third meals. Early blood meals were detected clearly within later blood meals as a delimited body of dark digested blood, heme (sometimes also with pink undigested blood), the presence of an associated pale pink-staining peritrophic plug, the presence and appearance of the peritrophic membrane surrounding the meals, and a physical space between meals; the first two characteristics were the most important. Development of the ovarian follicles including apparent dilatations was also observable using this histologic technique. The results of this study indicate that the rate of multiple feeding can be determined using histology. The technique would be useful in evaluating the blood feeding frequency of field-caught sand flies in endemic areas of leishmaniasis, bartonellosis, and phleboviruses.