Phlebotomus papatasi sand flies, the vectors of Leishmania major, differ genetically in their susceptibility to the pathogens. The costs of infection appear to be so great that selection against Leishmania-susceptible flies could presumably occur, unless susceptibility is compensated for by some advantage. Foci of P. papatasi-transmitted L. major are mainly found in arid habitats where seasonal stress of dehydration and heat reduces the quantity of sugar in plant leaves. The sand flies feed on these leaves and with the lack of essential sugar only a few survive long enough to deposit eggs and transmit Leishmania. This association suggested that susceptibility to L. major infection may be linked with advantageous tolerance of sugar deprivation. Here we show that desert sand flies, provided with excess sugar, became progressively resistant to infection. Selection for survival under sugar-poor conditions increased the susceptibility of parent and first-generation offspring by more than 2-fold. The leaves of plants, on which flies naturally feed, contain more sugar in irrigated than in arid habitats. About 85% of first-generation flies colonized from a desert habitat retain experimental Leishmania infection compared to 25% of offspring of flies from irrigated sites.

Binding to the midgut microvillar surface in the sandfly Phlebotomus papatasi is a prerequisite for successful development of Leishmania major within the gut of the vector. This paper describes a method for detecting microvillar-associated proteins which act as ligands for the parasite surface glycoconjugate lipophosphoglycan (LPG). Adhesion of LPG to midgut proteins was visualized by probing midgut extracts with LPG using a Western ligand blotting technique. Procyclic L. major LPG bound to a microvillar polypeptide band of 65 kDa (estimated in the non-reduced state) and bound variably to several lower molecular weight bands, probably degradation products or subunits of the primary binding polypeptides. Specificity of binding was confirmed by co-incubating biotinylated LPG with an LPG-specific mAb which resulted in a great reduction in binding.

The circumstances which permit the establishment of Leishmania infections in sandflies were investigated by altering the growth conditions for L. donovani parasites in the unsuitable vector Phlebotomus papatasi. Only 5·0% of the sandflies harboured a few parasites 3 days after feeding on promastigotes in defibrinated blood. Heparinized blood or the addition of trypsin inhibitor to the meals allowed persistence of infections (day 6) in 9·9% and 25·8% of the flies respectively. Meals of erythrocytes, saline and amastigotes produced 44·4% fly infection on day 6, while similar promastigote-initiated infections remained in 70·3% of the flies. Proteolytic activities in the guts of sandflies fed on the above meals without parasites, were the highest after defibrinated bloodmeals. Erythrocytes with saline decreased the maximal alkaline protease level from 20·8 U to 13·5 U/fly; that of trypsin from 3·9 U to 1·8 U/fly and that of the aminopeptidase from 5·5 U to 3·9 U/fly. After meals of heparinized blood, the maximal alkaline protease activity (12·0 U/fly) was also much lower than after defibrinated blood-feeding. The different diets which resulted in comparatively low enzymatic activities, including blood with trypsin inhibitor, also promoted the survival of infections. This implies that the proteolytic activity in the sandfly gut modulates the vector susceptibility.

This paper combines some of the results from three studies that explain the system by which the gut of Phlebotomus papatasi is maintained bacteria free. This system includes an antibacterial factor in the crop (diverticulum) and feeding behaviour which determines selective routing of meals. When feeding by piercing animal skin or plant tissue, in a ‘blood feeding’ mode, the uncontaminated meals of both sources enter the midgut directly. Free-solution sugar-meals which may contain contaminants, enter the crop where they mix with the antibacterial agent. Feeding on plants is also described.