Multiplicity of infection (MOI) and genetic diversity of P. falciparum infections are important surrogate indicators for assessing malaria transmission intensity in different regions of endemicity. Determination of MOI and diversity of P. falciparum among asymptomatic carriers will enhance our understanding of parasite biology and transmission to mosquito vectors. This study examined the MOI and genetic diversity of P. falciparum parasite populations circulating in Mbita, a region characterized as one of the malaria hotspots in Kenya. The genetic diversity and multiplicity of P. falciparum infections in 95 asymptomatic school children (age 5–15 yrs.) residing in Mbita, western Kenya were assessed using 10 polymorphic microsatellite markers. An average of 79.69% (Range: 54.84–95.74%) of the isolates analysed in this study were polyclonal infections as detected in at least one locus. A high mean MOI of 3.39 (Range: 2.24–4.72) and expected heterozygosity (He) of 0.81 (Range: 0.57–0.95) was reported in the study population. The analysed samples were extensively polyclonal infections leading to circulation of highly genetically diverse parasite populations in the study area. These findings correlated with the expectations of high malaria transmission intensity despite scaling up malaria interventions in the area thereby indicating the need for a robust malaria interventions particularly against asymptomatic carriers in order to attain elimination in the region.

Background: Asymptomatic Plasmodium falciparum gametocyte carriers are reservoirs for sustaining transmission in malaria endemic regions. Gametocyte presence in the host peripheral blood is a predictor of capacity to transmit malaria. However, it does not always directly translate to mosquito infectivity. Factors that affect mosquito infectivity include, gametocyte sex-ratio and density, multiplicity of infection (MOI), and host and vector anti-parasite immunity. We assess the prevalence of gametocyte carriage and some of its associated risk factors among asymptomatic schoolchildren in Western Kenya and to further analyse the association between gametocyte density, multiplicity of infection (MOI) and mosquito infection prevalence. Methods: P. falciparum parasite infections were detected by RDT (Rapid Diagnostic Test) and microscopy among schoolchildren (5-15 years old). Blood from 37 microscopy positive gametocyte carriers offered to laboratory reared An. gambiae s.l. mosquitoes. A total of 3395 fully fed mosquitoes were screened for Plasmodium sporozoites by ELISA. P. falciparum was genotyped using 10 polymorphic microsatellite markers. The association between MOI and gametocyte density and mosquito infection prevalence was investigated. Results: A significantly higher prevalence of P. falciparum infection was found in males 31.54% (764/2422) (p-value < 0.001) compared to females 26.72% (657/2459). The microscopic gametocyte prevalence among the study population was 2% (84/4881). Children aged 5-9 years have a higher prevalence of gametocyte carriage (odds ratios = 2.1 [95% CI = 1.3–3.4], P = 0.002) as compared to children aged 10-15 years. After offering gametocyte positive blood to An. gambiae s.l. by membrane feeding assay, our results indicated that 68.1% of the variation in mosquito infection prevalence was accounted for by gametocyte density and MOI (R-SQR. = 0.681, p < 0.001). Conclusions: We observed a higher risk of gametocyte carriage among the younger children (5-9 years). Gametocyte density and MOI significantly predicted mosquito infection prevalence.

Background: Malaria is a major public health threat in sub-Saharan Africa. Asymptomatic Plasmodium falciparum gametocyte carriers are potential infectious reservoirs for sustaining transmission in many malaria endemic regions. The aim of the study was to assess the prevalence of gametocyte carriage and some of its associated risk factors among asymptomatic schoolchildren in Western Kenya and further analyse the association between gametocyte density, multiplicity of infection (MOI) and mosquito infection prevalence. Methods: Rapid diagnostic tests were used to screen for P. falciparum parasite infection among schoolchildren (5-15 years old) and the results were verified using microscopy. Microscopy positive gametocyte carriers were selected to feed laboratory reared An. gambiae s.l. mosquitoes using membrane feeding method. Genomic DNA was extracted from dry blood spot samples and P. falciparum populations were genotyped using 10 polymorphic microsatellite markers. Assessment of the association between MOI and gametocyte density and mosquito infection prevalence was conducted. Results: A significantly higher prevalence of P. falciparum infection was found in males 31.54% (764/2422) (p-value < 0.001) compared to females 26.72% (657/2459). The microscopy gametocyte prevalence among the study population was 2% (84/4881). Children aged 5-9 years have a higher prevalence of gametocyte carriage (odds ratios = 2.1 [95% CI = 1.3–3.4], P = 0.002) as compared to children aged 10-15 years. After challenging An. gambiae s.l. by membrane feeding assay on gametocyte positive patient blood, our results indicate that 68.1% of the variation in mosquito infection prevalence is accounted for by gametocyte density and MOI (R-SQR. = 0.681, p < 0.001). Conclusions: Age was a significant risk factor for gametocyte carriage, as indicated by the higher risk of gametocyte carriage among the younger children (5-9 years). Gametocyte density and MOI statistically significantly predicted mosquito infection prevalence. Both of the variables added significantly to the prediction (p < 0.05).

African trypanosomes evade host immune clearance by antigenic variation, causing persistent infections in humans and animals. These parasites express a homogeneous surface coat of variant surface glycoproteins (VSGs). They transcribe one out of hundreds of VSG genes at a time from telomeric expression sites (ESs) and periodically change the VSG expressed by transcriptional switching or recombination. The mechanisms underlying the control of VSG switching and its developmental silencing remain elusive. We report that telomeric ES activation and silencing entail an on/off genetic switch controlled by a nuclear phosphoinositide signaling system. This system includes a nuclear phosphatidylinositol 5-phosphatase (PIP5Pase), its substrate PI(3,4,5)P3, and the repressor-activator protein 1 (RAP1). RAP1 binds to ES sequences flanking VSG genes via its DNA binding domains and represses VSG transcription. In contrast, PI(3,4,5)P3 binds to the N-terminus of RAP1 and controls its DNA binding activity. Transient inactivation of PIP5Pase results in the accumulation of nuclear PI(3,4,5)P3, which binds RAP1 and displaces it from ESs, activating transcription of silent ESs and VSG switching. The system is also required for the developmental silencing of VSG genes. The data provides a mechanism controlling reversible telomere silencing essential for the periodic switching in VSG expression and its developmental regulation.

Background: Malaria is a major public health threat in sub-Saharan Africa. Asymptomatic P. falciparum gametocyte carriers are potential infectious reservoirs for sustaining transmission in many malaria endemic regions. The aim of the study was to assess the prevalence of gametocyte carriage and some of its associated risk factors among asymptomatic schoolchildren (age 5-15 years) in Mbita, western Kenya and further analyse the association between gametocyte density, multiplicity of infection (MOI) and mosquito infectivity. Methods: Rapid diagnostic test (RDT) was used to screen for P. falciparum parasite infection among asymptomatic schoolchildren (5-15 years old) residing in Mbita, Western Kenya and the results were further verified using microscopy. Participants positive for P. falciparum infection were further screened for gametocyte carriage and those positive were used to feed laboratory reared An. gambiae s.s. mosquitoes using membrane feeding method. Genomic DNA was extracted from dry blood spots (DBS) samples and P. falciparum populations were genotyped using 10 polymorphic microsatellite markers. Assessment of the association between MOI and gametocyte density and mosquito infection rates was conducted. Results: The prevalence of Plasmodium falciparum infection among the study population was 29.1%. A significantly higher P. falciparum infection was found among the male gender (n=764, p -value < 0.05) compared to the females (n=657). Gametocyte prevalence among the study population was 2%. Children (5-9 years) have a higher risk of gametocyte carriage (Odd Ratios = 2.1 [95% CI = 1.3–3.4], P = 0.002). Our results indicate a significant and positive combined effect of gametocyte density and MOI on mosquito infection rate (R = 0.825, p < 0.0001). Conclusion: The study reports a relatively stable year-round gametocyte carriage among the study population. This pattern of gametocyte carriage signals the role of schoolchildren in maintaining malaria transmission in the study area. A strong positive correlation was found between gametocyte density, multiplicity of infection and mosquito infectivity.