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Reports suggest non-falciparum species are an underappreciated cause of malaria in sub-Saharan Africa but their epidemiology is ill-defined, particularly in highly malaria-endemic regions. We estimated incidence and prevalence of PCR-confirmed non-falciparum and Plasmodium falciparum malaria infections within a longitudinal study conducted in Kinshasa, Democratic Republic of Congo (DRC) between 2015-2017. Children and adults were sampled at biannual household surveys and routine clinic visits. Among 9,089 samples from 1,565 participants, incidences of P. malariae, P. ovale spp., and P. falciparum infections by 1-year were 7.8% (95% CI: 6.4%-9.1%), 4.8% (95% CI: 3.7%-5.9%) and 57.5% (95% CI: 54.4%-60.5%), respectively. Non-falciparum prevalences were higher in school-age children, rural and peri-urban sites, and P. falciparum co-infections. P. falciparum remains the primary driver of malaria in the DRC, though non-falciparum species also pose an infection risk. As P. falciparum interventions gain traction in high-burden settings, continued surveillance and improved understanding of non-falciparum infections are warranted., (© 2023. Springer Nature Limited.)

Introduction: insecticide-treated nets (ITNs) remain the mainstay of malaria vector control in the Democratic Republic of Congo. However, insecticide resistance of malaria vectors threatens their effectiveness. Entomological inoculation rates and insecticide susceptibility in Anopheles gambiae s.l. were evaluated before and after mass distribution of ITNs in Bandundu City for possible occurrence of resistance., Methods: a cross-sectional study was conducted from 15 th July 2015 to 15 th June 2016. Adult mosquitoes were collected using pyrethrum spray catches and human landing catches and identified to species level and tested for the presence of sporozoites. Bioassays were carried out before and after distribution of ITNs to assess the susceptibility of adult mosquitoes to insecticides. Synergist bioassays were also conducted and target site mutations assessed using Polymerase chain reaction (PCR)., Results: a total of 1754 female An. gambiae s.l. were collected before and after deployment of ITNs. Fewer mosquitoes were collected after the distribution of ITNs. However, there was no significant difference in sporozoite rates or the overall entomological inoculation rate before and after the distribution of ITNs. Test-mosquitoes were resistant to deltamethrin, permethrin, and Dichlorodiphenyltrichloroethane but susceptible to bendiocarb. Pre-exposure of mosquitoes to Piperonyl butoxide increased their mortality after exposure to permethrin and deltamethrin. The frequency of the Kinase insert domain receptor (kdr)-West gene increased from 92 to 99% before and after the distribution of nets, respectively., Conclusion: seasonal impacts could be a limiting factor in the analysis of these data; however, the lack of decrease in transmission after the distribution of new nets could be explained by the high-level of resistance to pyrethroid., Competing Interests: The authors declare no competing interests., (Copyright: Emery Metelo-Matubi et al.)

Background: The Democratic Republic of the Congo (DRC) remains one of the countries most impacted by malaria despite decades of control efforts, including multiple mass insecticide treated net (ITN) distribution campaigns. The multi-scalar and complex nature of malaria necessitates an understanding of malaria risk factors over time and at multiple levels (e.g., individual, household, community). Surveillance of households in both rural and urban settings over time, coupled with detailed behavioral and geographic data, enables the detection of seasonal trends in malaria prevalence and malaria-associated behaviors as well as the assessment of how the local environments within and surrounding an individual's household impact malaria outcomes., Methods: Participants from seven sites in Kinshasa Province, DRC were followed for over two years. Demographic, behavioral, and spatial information was gathered from enrolled households. Malaria was assessed using both rapid diagnostic tests (RDT) and polymerase chain reaction (PCR) and seasonal trends were assessed. Hierarchical regression modeling tested associations between behavioral and environmental factors and positive RDT and PCR outcomes at individual, household and neighborhood scales., Results: Among 1591 enrolled participants, malaria prevalence did not consistently vary seasonally across the sites but did vary by age and ITN usage. Malaria was highest and ITN usage lowest in children ages 6-15 years across study visits and seasons. Having another member of the household test positive for malaria significantly increased the risk of an individual having malaria [RDT: OR = 4.158 (2.86-6.05); PCR: OR = 3.37 (2.41-4.71)], as did higher malaria prevalence in the 250 m neighborhood around the household [RDT: OR = 2.711 (1.42-5.17); PCR: OR = 4.056 (2.3-7.16)]. Presence of water within close proximity to the household was also associated with malaria outcomes., Conclusions: Taken together, these findings suggest that targeting non-traditional age groups, children >5 years old and teenagers, and deploying household- and neighborhood-focused interventions may be effective strategies for improving malaria outcomes in high-burden countries like the DRC., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The majority of Plasmodium falciparum malaria diagnoses in Africa are made using rapid diagnostic tests (RDTs) that detect histidine-rich protein 2. Increasing reports of false-negative RDT results due to parasites with deletions of the pfhrp2 and/or pfhrp3 genes (pfhrp2/3) raise concern about existing malaria diagnostic strategies. We previously identified pfhrp2-negative parasites among asymptomatic children in the Democratic Republic of the Congo (DRC), but their impact on diagnosis of symptomatic malaria is unknown. We performed a cross-sectional study of false-negative RDTs in symptomatic subjects in 2017. Parasites were characterized by microscopy; RDT; pfhrp2/3 genotyping and species-specific PCR assays; a bead-based immunoassay for Plasmodium antigens; and/or whole-genome sequencing. Among 3627 symptomatic subjects, 427 (11.8%) had RDT-/microscopy + results. Parasites from eight (0.2%) samples were initially classified as putative pfhrp2/3 deletions by PCR, but antigen testing and whole-genome sequencing confirmed the presence of intact genes. 56.8% of subjects had PCR-confirmed malaria. Non-falciparum co-infection with P. falciparum was common (13.2%). Agreement between PCR and HRP2-based RDTs was satisfactory (Cohen's kappa = 0.66) and superior to microscopy (0.33). Symptomatic malaria due to pfhrp2/3-deleted P. falciparum was not observed. Ongoing HRP2-based RDT use is appropriate for the detection of falciparum malaria in the DRC.