IntroductionIn the era of malaria elimination and eradication, drug-based and vaccine-based approaches to reduce malaria transmission are receiving greater attention. Such interventions require assays that reliably measure the transmission of Plasmodium from humans to Anopheles mosquitoes.MethodsWe compared two commonly used mosquito feeding assay procedures: direct skin feeding assays and membrane feeding assays. Three conditions under which membrane feeding assays are performed were examined: assays with i) whole blood, ii) blood pellets resuspended with autologous plasma of the gametocyte carrier, and iii) blood pellets resuspended with heterologous control serum.Results930 transmission experiments from Cameroon, The Gambia, Mali and Senegal were included in the analyses. Direct skin feeding assays resulted in higher mosquito infection rates compared to membrane feeding assays (odds ratio 2.39, 95% confidence interval 1.94–2.95) with evident heterogeneity between studies. Mosquito infection rates in membrane feeding assays and direct skin feeding assays were strongly correlated (p<0.0001). Replacing the plasma of the gametocyte donor with malaria naïve control serum resulted in higher mosquito infection rates compared to own plasma (OR 1.92, 95% CI 1.68–2.19) while the infectiousness of gametocytes may be reduced during the replacement procedure (OR 0.60, 95% CI 0.52–0.70).ConclusionsDespite a higher efficiency of direct skin feeding assays, membrane feeding assays appear suitable tools to compare the infectiousness between individuals and to evaluate transmission-reducing interventions. Several aspects of membrane feeding procedures currently lack standardization; this variability makes comparisons between laboratories challenging and should be addressed to facilitate future testing of transmission-reducing interventions.

Malaria transmission in Africa is a dynamic and complex system that is so far superficially understood. Further knowledge is required to improve control of the disease. In the present report, we highlight the contribution of the so-called "secondary" malaria vectors to the overall parasite transmission intensity in several sites across Cameroon, through a retrospective analysis of surveys from the Organisation de Coordination pour la lutte Contre les Endémies en Afrique Centrale database. In total, 48,490 female anophelines belonging to 21 different species were collected between October 1998 and March 2003. Anopheles gambiae Giles, Anopheles arabiensis Patton, Anopheles funestus Giles, Anopheles nili (Theobald), and Anopheles moucheti Evans represented 89% of the total anopheline fauna. Beside these major vectors, malaria parasites or their circumsporozoite proteins were found in nine secondary malaria vectors: Anopheles ovengensis Awono-Ambene et al., Anopheles carnevalei Brunhes et al., Anopheles coustani Laveran, Anopheles hancocki Edwards, Anopheles marshallii (Theobald), Anopheles paludis Theobald, Anopheles pharoensis Theobald, Anopheles wellcomei Theobald, and Anopheles ziemanni Grtünberg. The mean infection rate of secondary vectors (1.36%) was significantly (P < 0.001) lower than that of major vectors (3.08%). An. pharoensis and An. ovengensis were repeatedly found infected by Plasmodium falciparum Welch and contributed substantially to the total malaria transmission intensity in some areas where they were abundant. Both species have strong exophilic and/or exophagic habits such that they might elude vector control directed against endophilic and endophagic malaria vectors.

Insectary-reared Anopheles gambiae were experimentally fed with the blood of 90 naturally infected human volunteers carrying gametocytes of Plasmodium falciparum. At least one mosquito was successfully infected in 74% of experiments. The probability that a gametocyte carrier was infective, the probability that a mos quito became infected, and the number of oocysts harboured were related to gametocyte density. The mean oroportion of male gametocytes was 0*217 (i.e., 3-6 females for every male). Sex ratios differed significantly between gametocyte carriers. Variation in sex ratio was not related to the probability that a gametocyte car rier was infective. Among infective people whose sex ratio estimates were based on a reasonable number of gametocytes, sex ratio significantly predicted the proportion of infected mosquitoes and mean oocyst load, with infectivity rising as the proportion of male gametocytes increased towards 50%. There was no indica tion that infectivity reached a peak at some intermediate sex ratio, as would be expected if random mating of gametes was the primary determinant of fertilization success. These results raise 2 interesting questions: why should higher sex ratios be more infective, and why is the observed population sex ratio lower than that which produces the greatest infectivity?