Sandfly females, while feeding on the host, excrete urine to concentrate proteins of the bloodmeal and restore weight and water balance. This process, analogous to prediuresis in mosquitoes, was observed in 100% of Phlebotomus papatasi (Scopoli) and 85% of P. duboscqi Neveu-Lemaire females studied. Individual females, however, differed in duration of prediuresis and in the number of ejected urine droplets. In both species the prediuresis generally started 1-2 min after the commencement of feeding and the variation in urine production was positively correlated with the length of feeding. The first one or two droplets were opaque whitish while the remaining ones were clear. Erythrocytes were found sporadically in first droplets of some females. Representative prediuresis in P. duboscqi included 27 droplets, i.e., about 325 nl urine in total, ejected during 8 min of feeding. The study revealed prediuresis in P. papatasi and P. duboscqi as a regular physiological process which may have consequences in transmission of infective diseases.
The Leishmania metalloproteinase GP63 has been reported to play important roles mainly in resistance of promastigotes to complement-mediated lysis and in interaction with macrophage receptors. On the other hand, its function in insect vectors is still unclear. We compared the structure and dosage of gp63 genes and the activity of GP63 in Leishmania major Yakimoff et Schokhor strains and lines differing in virulence for mice and ability to develop in sand flies. The results demonstrate considerable variability in amount and proteolytical activity of GP63 among L. major strains although genomic changes in the gp63 locus were not found. Attenuated LV561/AV line showed low amount and low enzymatic activity of GP63. Serial passages of attenuated parasites through either Phlebotomus duboscqi Neveu-Lemaire or through mice led to a recovery of GP63 proteolytical activity to the level present in virulent LV561/V line. Overexpression of GP63 was found in two L. major strains (L119, Neal) with defective lipophosphoglycan (LPG); both these strains were capable to cause mice infection but unable to survive and multiply in sand flies. Differences were found also in karyotypes and in amount of minichromosomes amplified in some lines of the LV561 strain. The results suggest that parasite virulence is not simply correlated with the activity of GP63; however, this enzyme plays a significant role in association with other surface molecules, especially LPG. Overexpression of GP63 can compensate LPG defect in the vertebrate host but in sand flies both molecules fulfil quite different functions and the defect in LPG is lethal for the parasite. On the other hand, linear minichromosomes of about 200 kb found in some lines of the LV561 strain are associated with development in vitro and in the vector but they are not essential for the infection of the vertebrate host.