Plant-Mediated Effects on Mosquito Capacity to Transmit Human Malaria.

The ecological context in which mosquitoes and malaria parasites interact has received little attention, compared to the genetic and molecular aspects of malaria transmission. Plant nectar and fruits are important for the nutritional ecology of malaria vectors, but how the natural diversity of plant-derived sugar sources affects mosquito competence for malaria parasites is unclear. To test this, we infected Anopheles coluzzi, an important African malaria vector, with sympatric field isolates of Plasmodium falciparum, using direct membrane feeding assays. Through a series of experiments, we then examined the effects of sugar meals from Thevetia neriifolia and Barleria lupilina cuttings that included flowers, and fruit from Lannea microcarpa and Mangifera indica on parasite and mosquito traits that are key for determining the intensity of malaria transmission. We found that the source of plant sugar meal differentially affected infection prevalence and intensity, the development duration of the parasites, as well as the survival and fecundity of the vector. These effects are likely the result of complex interactions between toxic secondary metabolites and the nutritional quality of the plant sugar source, as well as of host resource availability and parasite growth. Using an epidemiological model, we show that plant sugar source can be a significant driver of malaria transmission dynamics, with some plant species exhibiting either transmission-reducing or -enhancing activities.

Experimental study of the relationship between Plasmodium gametocyte density and infection success in mosquitoes; implications for the evaluation of malaria transmission-reducing interventions.

The evaluation of transmission reducing interventions (TRI) to control malaria widely uses membrane feeding assays. In such assays, the intensity of Plasmodium infection in the vector might affect the measured efficacy of the candidates to block transmission. Gametocyte density in the host blood is a determinant of the infection success in the mosquito, however, uncertain estimates of parasite densities and intrinsic characteristics of the infected blood can induce variability. To reduce this variation, a feasible method is to dilute infectious blood samples. We describe the effect of diluting samples of Plasmodium-containing blood samples to allow accurate relative measures of gametocyte densities and their impact on mosquito infectivity and TRI efficacy. Natural Plasmodium falciparum samples were diluted to generate a wide range of parasite densities, and fed to Anopheles coluzzii mosquitoes. This was compared with parallel dilutions conducted on Plasmodium berghei infections. We examined how blood dilution influences the observed blocking activity of anti-Pbs28 monoclonal antibody using the P. berghei/Anopheles stephensi system. In the natural species combination P. falciparum/An. coluzzii, blood dilution using heat-inactivated, infected blood as diluents, revealed positive near linear relationships, between gametocyte densities and oocyst loads in the range tested. A similar relationship was observed in the P. berghei/An. stephensi system when using a similar dilution method. In contrast, diluting infected mice blood with fresh uninfected blood dramatically increases the infectiousness. This suggests that highly infected mice blood contains inhibitory factors or reduced blood moieties, which impede infection and may in turn, lead to misinterpretation when comparing individual TRI evaluation assays. In the lab system, the transmission blocking activity of an antibody specific for Pbs28 was confirmed to be density-dependent. This highlights the need to carefully interpret evaluations of TRI candidates, regarding gametocyte densities in the P. berghei/An. stephensi system.

Interplay between Plasmodium infection and resistance to insecticides in vector mosquitoes.

Despite its epidemiological importance, the impact of insecticide resistance on vector-parasite interactions and malaria transmission is poorly understood. Here, we explored the impact of Plasmodium infection on the level of insecticide resistance to dichlorodiphenyltrichloroethane (DDT) in field-caught Anopheles gambiae sensu stricto homozygous for the kdr mutation. Results showed that kdr homozygous mosquitoes that fed on infectious blood were more susceptible to DDT than mosquitoes that fed on noninfectious blood during both ookinete development (day 1 after the blood meal) and oocyst maturation (day 7 after the blood meal) but not during sporozoite invasion of the salivary glands. Plasmodium falciparum infection seemed to impose a fitness cost on mosquitoes by reducing the ability of kdr homozygous A. gambiae sensu stricto to survive exposure to DDT. These results suggest an interaction between Plasmodium infection and the insecticide susceptibility of mosquitoes carrying insecticide-resistant alleles. We discuss this finding in relation to vector control efficacy.

Studying fitness cost of Plasmodium falciparum infection in malaria vectors: validation of an appropriate negative control.

BACKGROUND: The question whether Plasmodium falciparum infection affects the fitness of mosquito vectors remains open. A hurdle for resolving this question is the lack of appropriate control, non-infected mosquitoes that can be compared to the infected ones. It was shown recently that heating P. falciparum gametocyte-infected blood before feeding by malaria vectors inhibits the infection. Therefore, the same source of gametocyte-infected blood could be divided in two parts, one heated, serving as the control, the other unheated, allowing the comparison of infected and uninfected mosquitoes which fed on exactly the same blood otherwise. However, before using this method for characterizing the cost of infection to mosquitoes, it is necessary to establish whether feeding on previously heated blood affects the survival and fecundity of mosquito females. METHODS: Anopheles gambiae M molecular form females were exposed to heated versus non-heated, parasite-free human blood to mimic blood meal on non-infectious versus infectious gametocyte-containing blood. Life history traits of mosquito females fed on blood that was heat-treated or not were then compared. RESULTS: The results reveal that heat treatment of the blood did not affect the survival and fecundity of mosquito females. Consistently, blood heat treatment did not affect the quantity of blood ingested. CONCLUSIONS: The study indicates that heat inactivation of gametocyte-infected blood will only inhibit mosquito infection and that this method is suitable for quantifying the fitness cost incurred by mosquitoes upon infection by P. falciparum.

Serological and molecular detection of Leishmania species in dog peripheral blood from Bobo-Dioulasso city, a confirmation of canine leishmaniasis enzootic area for Burkina Faso.

Canine leishmaniasis is increasingly reported worldwide and represent a threat to both animal and human health. In a previous pilot study conducted in Bobo-Dioulasso, the second town of Burkina Faso, we reported five cases of canine leishmaniasis. With the perspective of a One Health action plan, and in the context of increasing urbanization, this study aimed to provide new information on Leishmania spp in dogs in this city. A cross-sectional survey was carried out from May to August 2018 in six districts of the city in order to record clinical and biological data from domestic dogs randomly selected per district. Blood samples were collected into EDTA tubes (4-5 mL), treated and stored at -20 °C until further analyses. The infection status of the dogs was performed by serological tests using plasma, and real time-PCR (RT-PCR) to detect Leishmania parasites using buffy coats. Nested PCR was used for typing the Leishmania species in dogs which were found to be RT-PCR positive. A total of 147 dogs were examined clinically and sampled for blood collection, including 53.7% females and 46.3% of males with a median age of 3 years. The seroincidence of Leishmania parasites within this dog population was 4.76% (95% CI:2.26-9.72). The incidence of Leishmania was 10.88% (95% CI: 6.73-17.11) by RT-PCR which was significantly more sensitive (p = 0,047) and a fair concordance was observed between both tests (Kappa = 0.39, p < 0.001). The characterization of Leishmania species revealed that L. major was circulating in this domestic dog population. Our results confirmed the persistence of zoonotic circulation of Leishmania parasites such as L. major currently in Bobo-Dioulasso city and highlight the need for targeted interventions in order to control transmission of leishmaniasis in this region.

No evidence for manipulation of Anopheles gambiae, An. coluzzii and An. arabiensis host preference by Plasmodium falciparum.

Whether malaria parasites can manipulate mosquito host choice in ways that enhance parasite transmission toward suitable hosts and/or reduce mosquito attraction to unsuitable hosts (i.e. specific manipulation) is unknown. To address this question, we experimentally infected three species of mosquito vectors with wild isolates of the human malaria parasite Plasmodium falciparum, and examined the effects of immature and mature infections on mosquito behavioural responses to combinations of calf odour, human odour and outdoor air using a dual-port olfactometer. Regardless of parasite developmental stage and mosquito species, P. falciparum infection did not alter mosquito activation rate or their choice for human odours. The overall expression pattern of host choice of all three mosquito species was consistent with a high degree of anthropophily, with infected and uninfected individuals showing higher attraction toward human odour over calf odour, human odour over outdoor air, and outdoor air over calf odour. Our results suggest that, in this system, the parasite may not be able to manipulate the early long-range behavioural steps involved in the mosquito host-feeding process. Future studies are required to test whether malaria parasites can modify their mosquito host choice at a shorter range to enhance transmission.

Prevalence of intestinal opportunistic parasites infections in the University hospital of Bobo-Dioulasso, Burkina Faso.

BACKGROUND: Gastrointestinal parasites infections are widespread in Africa and their prevalence infections vary from country to country. This study aimed at assessing the prevalence of opportunistic intestinal parasites infection and other gastrointestinal parasites infection among patients attending the laboratory of Parasitology and Mycology of the University Hospital Souro Sanou of Bobo-Dioulasso. METHODS: A hospital cross-sectional based study was conducted from April to August, 2012. Participants were persons whom parasitological examination of stools has been prescribed by a clinician. The stools examination methods included direct wet saline examination, lugol's iodine staining technique, formol-ether concentration and modified Ziehl-Neelsen staining. We recorded age and sex information for each patient. RESULTS: The overall prevalence of intestinal parasite infections was 65.3 % (190/291). Majority of the parasitic infections was waterborne (64.3 %) consisting of high prevalence of Cryptosporidium sp. (26.5 %) and Entamoeba histolytica/dispar (23.4 %). The prevalence of opportunistic parasites was 28.9 % and Cryptosporidium sp. was the most prevalent species followed by Blastocystis sp. (1.0 %), Cyclospora sp. (0.7 %) and Isospora belli (0.7 %). The prevalence of intestinal helminthes was 1.7 %. CONCLUSIONS: The prevalence of intestinal parasitism in general remains high in Bobo-Dioulasso requiring the establishment of adequate diagnostic techniques, treatment and prevention.

Comparison of field-based xenodiagnosis and direct membrane feeding assays for evaluating host infectiousness to malaria vector Anopheles gambiae.

Several techniques are currently being used to study host infectiousness to mosquitoes, including the experimental possibility of laboratory reared mosquitoes acquiring infections through membrane feeders or directly on host skin. Here, the relative performance of the laboratory-based membrane feeding method (DMFA) and the field-based xenodiagnosis (XD) of malaria infectious hosts using wild Anopheles mosquitoes were compared. A cross-sectional survey involving a sample of 70 children (aged 3-12 years) living in a malaria endemic area in Western Burkina Faso, was carried out to measure their infectiousness to Anopheles mosquitoes using two approaches. The first approach used the xenodiagnostic procedure in which children were exposed to mosquito bites overnight, being sleeping individually in different sentinel huts from 6 pm to 6 am (4 nights per child). Anopheles sp that had acquired blood-meal on each child were subsequently collected early in the morning, and examined for Plasmodium falciparum oocyst infection on day 7 post-feeding. In the second approach, the infectiousness of the same children was estimated by whole-blood membrane feeding procedure using F0 An. gambiae s.l. that emerged from field-collected larvae cohorts. In the DMFA, 41.4% of the children successfully infected at least one mosquito with the mean oocyst prevalence of only 4.6±1.1% in the 2171 mosquitoes that were examined (mean oocyst intensity: 2.0±(std error of mean) 0.3 oocysts per infected midgut). Comparatively 78.6% of children yielded oocysts infection in mosquitoes during the XD approach (Chi square=20.11, df=1; p<0.001), with a mean rate of 19.6±2.0 in the 3752 wild caught mosquitoes (mean intensity: 3.93±0.2 oocysts per infected mosquito). The DMFA failed to reveal a portion (n=26) of infectious individuals that were sharply evidenced by the XD, particularly at low gametocyte densities or at levels that could not be detected by the classical microscopic examination of blood smears. As opposed to the resource consuming DMFA, which is often mined by technical constraints, using the XD method could be an advantage in experimental investigations of host infectiousness in areas where anopheline species cannot be conveniently reared for the experimental studies. Ethical aspects of this approach, mainly related to exposure of the human subjects to potentially infectious mosquito bites are discussed.

Genetic variation in human HBB is associated with Plasmodium falciparum transmission.

Genetic factors are known to have a role in determining susceptibility to infectious diseases, although it is unclear whether they may also influence host efficiency in transmitting pathogens. We examine variants in HBB that have been shown to be protective against malaria and test whether these are associated with the transmission of the parasite from the human host to the Anopheles vector. We conducted cross-sectional malariological surveys on 3,739 human subjects and transmission experiments involving 60 children and 6,446 mosquitoes in Burkina Faso, West Africa. Protective hemoglobins C (HbC, beta6Glu-->Lys) and S (beta6Glu-->Val) are associated with a twofold in vivo (odds ratio 2.17, 95% CI 1.57-3.01, P = 1.0 x 10(-6)) and a fourfold ex vivo (odds ratio 4.12, 95% CI 1.90-9.29, P = 7.0 x 10(-5)) increase of parasite transmission from the human host to the Anopheles vector. This provides an example of how host genetic variation may influence the transmission dynamics of an infectious disease.