Gametocytes infectiousness to mosquitoes: variable selection using random forests, and zero inflated models

Malaria control strategies aiming at reducing disease transmission intensity may impact both oocyst intensity and infection prevalence in the mosquito vector. Thus far, mathematical models failed to identify a clear relationship between Plasmodium falciparum gametocytes and their infectiousness to mosquitoes. Natural isolates of gametocytes are genetically diverse and biologically complex. Infectiousness to mosquitoes relies on multiple parameters such as density, sex-ratio, maturity, parasite genotypes and host immune factors. In this article, we investigated how density and genetic diversity of gametocytes impact on the success of transmission in the mosquito vector. We analyzed data for which the number of covariates plus attendant interactions is at least of order of the sample size, precluding usage of classical models such as general linear models. We then considered the variable importance from random forests to address the problem of selecting the most influent variables. The selected covariates were assessed in the zero inflated negative binomial model which accommodates both over-dispersion and the sources of non infected mosquitoes. We found that the most important covariates related to infection prevalence and parasite intensity are gametocyte density and multiplicity of infection

Can field-based mosquito feeding assays be used for evaluating transmission-blocking interventions?

A recent meta-analysis of mosquito feeding assays to determine the Plasmodium falciparum transmission potential of naturally infected gametocyte carriers highlighted considerable variation in transmission efficiency between assay methodologies and between laboratories. This begs the question as to whether mosquito feeding assays should be used for the evaluation of transmission-reducing interventions in the field and whether these field-based mosquito assays are currently standardized sufficiently to enable accurate evaluations. Here, we address biological and methodological reasons for the observed variations, discuss whether these preclude the use of field-based mosquito feeding assays in field evaluations of transmission-blocking interventions, and propose how we can maximize the precision of estimates. Altogether, we underscore the significant advantages of field-based mosquito feeding assays in basic malaria research and field trials

Gametocytes infectiousness to mosquitoes: variable selection using random forests, and zero inflated models

Malaria control strategies aiming at reducing disease transmission intensity may impact both oocyst intensity and infection prevalence in the mosquito vector. Thus far, mathematical models failed to identify a clear relationship between Plasmodium falciparum gametocytes and their infectiousness to mosquitoes. Natural isolates of gametocytes are genetically diverse and biologically complex. Infectiousness to mosquitoes relies on multiple parameters such as density, sex-ratio, maturity, parasite genotypes and host immune factors. In this article, we investigated how density and genetic diversity of gametocytes impact on the success of transmission in the mosquito vector. We analyzed data for which the number of covariates plus attendant interactions is at least of order of the sample size, precluding usage of classical models such as general linear models. We then considered the variable importance from random forests to address the problem of selecting the most influent variables. The selected covariates were assessed in the zero inflated negative binomial model which accommodates both over-dispersion and the sources of non infected mosquitoes. We found that the most important covariates related to infection prevalence and parasite intensity are gametocyte density and multiplicity of infection

SNP discovery and molecular evolution in Anopheles gambiae, with special emphasis on innate immune system

<p>Abstract</p> <p>Background</p> <p><it>Anopheles </it>innate immunity affects <it>Plasmodium </it>development and is a potential target of innovative malaria control strategies. The extent and distribution of nucleotide diversity in immunity genes might provide insights into the evolutionary forces that condition pathogen-vector interactions. The discovery of polymorphisms is an essential step towards association studies of susceptibility to infection.</p> <p>Results</p> <p>We sequenced coding fragments of 72 immune related genes in natural populations of <it>Anopheles gambiae </it>and of 37 randomly chosen genes to provide a background measure of genetic diversity across the genome. Mean nucleotide diversity (π) was 0.0092 in the <it>A. gambiae </it>S form, 0.0076 in the M form and 0.0064 in <it>A. arabiensis</it>. Within each species, no statistically significant differences in mean nucleotide diversity were detected between immune related and non immune related genes. Strong purifying selection was detected in genes of both categories, presumably reflecting strong functional constraints.</p> <p>Conclusion</p> <p>Our results suggest similar patterns and rates of molecular evolution in immune and non-immune genes in <it>A. gambiae</it>. The 3,214 Single Nucleotide Polymorphisms (SNPs) that we identified are the first large set of <it>Anopheles </it>SNPs from fresh, field-collected material and are relevant markers for future phenotype-association studies.</p

Concentration and purification by magnetic separation of the erythrocytic stages of all human Plasmodium species

International audienceBackground : Parasite concentration methods facilitate molecular, biochemical and immunologicalresearch on the erythrocytic stages of Plasmodium. In this paper, an adaptation of magnetic MACS®columns for the purification of human Plasmodium species is presented. This method was useful forthe concentration/purification of either schizonts or gametocytes.Results and conclusions : The magnetic removal of non-parasitized red blood cells (in vivo andin vitro) using magnetic columns (MACS) was evaluated. This easy-to-use technique enrichedschizonts and gametocytes from Plasmodium falciparum in vitro cultures with a very high degree ofpurity. In addition, all haemozoin-containing stages (schizonts and/or gametocytes) from theperipheral blood of infected patients could be concentrated using this method. This method isparticularly useful for the concentration of non-falciparum species, which do not grow in cultureand are otherwise difficult to obtain in large amounts

Plasmodium falciparum Produce Lower Infection Intensities in Local versus Foreign Anopheles gambiae Populations

Both Plasmodium falciparum and Anopheles gambiae show great diversity in Africa, in their own genetic makeup and population dynamics. The genetics of the individual mosquito and parasite are known to play a role in determining the outcome of infection in the vector, but whether differences in infection phenotype vary between populations remains to be investigated. Here we established two A. gambiae s.s. M molecular form colonies from Cameroon and Burkina Faso, representing a local and a foreign population for each of the geographical sites. Experimental infections of both colonies were conducted in Cameroon and Burkina Faso using local wild P. falciparum, giving a sympatric and allopatric vector-parasite combination in each site. Infection phenotype was determined in terms of oocyst prevalence and intensity for at least nine infections for each vector-parasite combination. Sympatric infections were found to produce 25% fewer oocysts per midgut than allopatric infections, while prevalence was not affected by local/foreign interactions. The reduction in oocyst numbers in sympatric couples may be the result of evolutionary processes where the mosquito populations have locally adapted to their parasite populations. Future research on vector-parasite interactions must take into account the geographic scale of adaptation revealed here by conducting experiments in natural sympatric populations to give epidemiologically meaningful results

Anopheles gambiae salivary protein expression modulated by wild Plasmodium falciparum infection: Highlighting of new antigenic peptides as candidates of An. gambiae bites

Background: Malaria is the major parasitic disease worldwide caused by Plasmodium infection. The objective of integrated malaria control programs is to decrease malaria transmission, which needs specific tools to be accurately assessed. In areas where the transmission is low or has been substantially reduced, new complementary tools have to be developed to improve surveillance. A recent approach, based on the human antibody response to Anopheles salivary proteins, has been shown to be efficient in evaluating human exposure to Anopheles bites. The aim of the present study was to identify new An. gambiae salivary proteins as potential candidate biomarkers of human exposure to P. falciparum-infective bites. Methods: Experimental infections of An. gambiae by wild P. falciparum were carried out in semi-field conditions. Then a proteomic approach, combining 2D-DIGE and mass spectrometry, was used to identify the overexpressed salivary proteins in infected salivary glands compared to uninfected An. gambiae controls. Subsequently, a peptide design of each potential candidate was performed in silico and their antigenicity was tested by an epitope-mapping technique using blood from individuals exposed to Anopheles bites. Results: Five salivary proteins (gSG6, gSG1b, TRIO, SG5 and long form D7) were overexpressed in the infected salivary glands. Eighteen peptides were designed from these proteins and were found antigenic in children exposed to the Anopheles bites. Moreover, the results showed that the presence of wild P. falciparum in salivary glands modulates the expression of several salivary proteins and also appeared to induce post-translational modifications. Conclusions: This study is, to our knowledge, the first that compares the sialome of An. gambiae both infected and not infected by wild P. falciparum, making it possible to mimic the natural conditions of infection. This is a first step toward a better understanding of the close interactions between the parasite and the salivary gland of mosquitoes. In addition, these results open the way to define biomarkers of infective bites of Anopheles, which could, in the future, improve the estimation of malaria transmission and the evaluation of malaria vector control tools. (Résumé d'auteur

Cancer incidence in the AGRICAN cohort study (2005-2011).

BACKGROUND: Numerous studies have been conducted among farmers, but very few of them have involved large prospective cohorts, and few have included a significant proportion of women and farm workers. Our aim was to compare cancer incidence in the cohort (overall, by sex, and by work on farm, occupational status and pesticide use) within the general population. METHODS: More than 180,000 participants in the AGRICAN cohort were matched to cancer registries to identify cancer cases diagnosed from enrolment (2005-2007) to 31st December 2011. We calculated standardized incidence ratios (SIRs) and 95% confidence intervals (95%CIs). RESULTS: Over the period, 11,067 incident cancer cases were identified (7304 men and 3763 women). Overall cancer incidence did not differ between the cohort and the general population. Moreover, SIRs were significantly higher for prostate cancer (SIR=1.07, 95%CI 1.03-1.11) and non-Hodgkin lymphoma (SIR=1.09, 95%CI 1.01-1.18) among men, skin melanoma among women (SIR=1.23, 95%CI 1.05-1.43) and multiple myeloma (men: SIR=1.38, 95%CI 1.18-1.62; women: SIR=1.26, 95%CI 1.02-1.54). In contrast, SIRs were lower for upper aerodigestive tract and respiratory cancers. Increase in risk was greater in male farm workers for prostate and lip cancer, in female farm workers for skin melanoma, and in male farm owners for multiple myeloma. Moreover, incidence of multiple myeloma and skin melanoma was higher among male and female pesticide users respectively. CONCLUSION: We found a decreased incidence for tobacco-related cancers and an increased incidence of prostate cancers, skin melanoma and multiple myeloma. Specific subgroups had a higher cancer incidence related to occupational status and pesticide use