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Sand fly salivary fluid contains numerous proteins that modulate host immune responses to infection and facilitate blood-feeding and establishing Leishmania infection. Salivary proteins are differentially expressed in adaptation to the host, the meal type and ecological factors. We report on the morphometrics and protein composition of salivary glands of colonised Phlebotomus papatasi and P. langeroni sand flies from Egypt. Female glands were dissected at day 1 (D1, unfed), day 2 (D2, sugar-fed), day 3 (D3, blood-fed) and day 7 (D7, blood-digested). The salivary glands are composed of two lobes: heterogeneous in P. papatasi and homogeneous in P. langeroni. Lobe sizes varied considerably with fly age and feeding state; D3 flies had the largest lobe sizes and protein content. The P. papatasi flies had larger lobes and higher protein content than the P. langeroni flies. The P. papatasi D1 flies had 15 protein bands that decreased in the D2, D3 and D7 flies to 10 bands in the Sinai flies and 9 bands in the Alexandria flies. All P. langeroni flies had 12 protein bands but with different intensities. The results reveal inter-specific variation between P. papatasi and P. langeroni, while no intra-specific variation between P. papatasi strains. These results increase our understanding of salivary gland protein composition and blood-feeding behaviour in Old World sand flies with implications for leishmaniasis epidemiology and control.

The Alexandria laboratory colony and five field populations of Phlebotomus papatasi (Scopoli) from Egypt were analyzed for genetic variation at 17 enzyme loci. The laboratory colony was characterized by a low level of genetric variation as measured by the average number of alleles per locus (A = 1.70 + or - 0.16) and the average expected heterozygosity (He = 0.06 + or - 0.02 ). Polymorphism was observed at 23.5% of the examined loci, and genotype frequencies at two loc (PGM, AK-2) were found to deviate slightly from the Hardy-Weinberg equilibrium. In contrast, the average number of alleles per locus for field populations ranged from A = 2.35 + or - 0.02 to 2.76 + or - 0.10, and He ranged from 0.15 + or - 0.03 to 0.21 + or - 0.05. All loci of field populations exhibited polymorphism, from 47.0 percent to 76.5%, and four to seven loci in each population were found to deviate from Hardy-Weinberg equilibrium. Deviation in both colonized and field populations were caused by heterozygote deficiency. Despite geographic isolation and some individual deviations from the Hardy-Weinberg equilibrium, no evidence of significant genetic difference was obtained for any of the populations sampled. Calculated indices of genetic distance and genetic identify for the five field populations showed minor variation but were collectively representative of a single genetically uniform population

Trials were conducted to determine the accuracy of separating the sympatric sand fly species Phlebotomus papatasi (Scopoli) and P. langeroni Nitzulescu by means of cellulose acetate enzyme electrophoresis. Malic enzyme, phosphoglucomutase, 6-phosphogluconate dehydrogenase, and fumarate hydratase were each evaluated in laboratory-reared and field-collected populations of the two species. Each of the four enzyme-based identifications was highly sensitive (greater than 97%) and specific (greater than 93%). Identifications based upon fumarate hydratase were in perfect agreement with morphological identifications, and evidence was obtained which indicates that this enzyme may be the most stable of the four enzymes tested. The application of enzyme-based vector identification is discussed in relation to classical and novel survey procedures for Leishmania promastigote detection in sand flies.

The dot enzyme-linked immunosorbent assay (dot-ELISA) was compared to the indirect immunofluorescent assay (IIFA) and radioimmunoassay (RIA) for the detection of malaria antibodies. Of 281 sera tested by dot-ELISA, 220 were from Ethiopia, 11 from the Sudan, and 50 from Egypt. A close correlation between the dot-ELISA and RIA results was observed in 92% of the 220 Ethiopian cases. Of the remainder, 6% gave positive RIA and negative dot-ELISA results, and 2% gave positive dot-ELISA and negative RIA results. Antibody titres determined by dot-ELISA and RIA were positively correlated in 10 of the 11 Sudanese cases tested by direct microscopical examination. The eleventh case was positive by dot-ELISA at 1:1000 dilution, but negative by RIA and direct examination. With the 50 Egyptian sera, the dot-ELISA results showed close correspondence to the IIFA results, but the dot-ELISA was 20–40 fold more sensitive than the IIFA. To test specificity, 62 samples from patients with 11 different diseases and conditions were examined by dot-ELISA. No malaria antibodies were detected in any of these or in sera from healthy controls. Dot-ELISA is a potentially useful method for sero-epidemiological studies of malaria.