Malaria diagnosis by direct observation of centrifuged samples of blood.

We compared the effectiveness of malaria diagnosis by means of direct observation of centrifuged blood with that by conventional examination of Giemsa stained blood-films in a malaria clinic in Ethiopia. A commercially available, modified hematological apparatus (the QBC tube) was used for centrifugation. Red blood cells infected with diverse stages of Plasmodium falciparum and P. vivax are lighter than noninfected cells and somewhat heavier than granulocytes; thus they can readily be detected by direct inspection of UV-illuminated tubes. About 10% of infections diagnosed by direct centrifugal microscopy in a clinical setting were not detected by conventional examination of stained thick films. Diagnosis by direct centrifugation appears to be at least 8 times as sensitive as conventional microscopy when applied to serially diluted samples of malaria-infected blood. Superior sensitivity, together with the one step, solid state nature of the direct centrifugal procedure, provides important advantages for malaria diagnosis.

Time and site of assembly of the peritrophic membrane of the mosquito Aedes aegypti.

We determined the time and site of secretion of the precursors of the peritrophic membrane (PM) in Aedes aegypti and when the structure is assembled. The fine structure of the developing membrane of blood-feed females was described, and the pattern of secretion of injected tritiated glucosamine analyzed autoradiographically. Immediately following blood feeding, ingested red cells rapidly become compressed, such that the surrounding plasma is extruded to the margin of the midgut contents. Thereby, ingested fluids form a narrow margin separating the blood mass from the midgut epithelium. By electron microscopy, the PM first becomes evident at about 4 to 8 h after blood is ingested, and the membrane attains mature texture by 12 h. The compacted mass of ingested erythrocytes seems to serve as a template for the forming structure. In contrast, tritiated glucosamine, injected into freshly engorged mosquitoes, begins to concentrate on the midgut microvilli by 2 h after feeding. By 8 h the label assumes the layered appearance that characterizes the fine structure of the mature membrane. In contrast to the prevailing concept that the PM of mosquitoes first assumes texture anteriorly immediately after blood is ingested, we find that this potential barrier to pathogen development forms no earlier than 4 h after feeding and that it is formed from precursors secreted along the entire length of the epithelium overlying the food mass.

Microfilarial perforation of the midgut of a mosquito.

To determine whether the midgut envelope of mosquitoes is disrupted by the passage of microfilariae, ultrastructural changes induced by microfilariae of Brugia malayi were observed in midguts of Aedes aegypti mosquitoes. Basal and apical plasma membranes were destroyed, disrupting the full depth of the midgut wall. Ingested ferritin lay against the gut wall, suggesting absence of the peritrophic membrane during penetration. Exsheathment of microfilariae appears to be enhanced by movement against the constricting midgut wall. It was concluded that particles present in the lumen of the gut may be disseminated passively to the hemocoel.