Article https://doi.org/10.1038/s41467-023-42190-w Epidemiology of Plasmodium malariae and Plasmodium ovale spp. in Kinshasa Province, Democratic Republic of Congo Rachel Sendor 1 , Kristin Banek 2 , Melchior M. Kashamuka 3 , Nono Mvuama 3 , Joseph A. Bala 3 , Marthe Nkalani 3 , Georges Kihuma 3 , Joseph Atibu 3 , Kyaw L. Thwai 2 , W. Matthew Svec 4 , Varun Goel 5,6 , Tommy Nseka 3 , Jessica T. Lin 2,7 , Jeffrey A. Bailey 8 , Michael Emch 1,5 , Margaret Carrel 9 , Jonathan J. Juliano 1,2,7 , Antoinette Tshefu 3,10 & Jonathan B. Parr 2,7,10 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. falci- parum 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 pre- valences 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. Reports of non-falciparum malaria caused by Plasmodium malariae and Plasmodium ovale spp. have increased across sub-Saharan Africa, where more than 95% of global malaria cases and deaths occur 1 . Plas- modium falciparum is the primary cause of malaria morbidity and mortality in the region. However, molecular surveys have confirmed co-circulating non-falciparum species, and suggest a rise in prevalence in regions where P. falciparum has declined 2–6 . Despite this, our understanding of infection risk and clinical burden posed by non- falciparum species is limited, particularly within regions of high P. falciparum transmission. The Democratic Republic of Congo (DRC) is one of 11 malaria “High Burden, High Impact” countries designated by the World Health Organization (WHO) 7 , indicating a critical need to understand the full landscape of malaria transmission and infection in the country. The Received: 6 April 2023 Accepted: 3 October 2023 Check for updates 1 Department of Epidemiology, Gillings School of Global Public Health, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA. 2 Institute for Global Health and Infectious Diseases, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA. 3 Ecole de Santé Publique, Faculté de Médecine, University of Kinshasa, Kinshasa, Democratic Republic of the Congo. 4 University of North Carolina at Chapel Hill, Chapel Hill, NC, USA. 5 Department of Geography, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA. 6 Carolina Population Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA. 7 Division of Infectious Diseases, School of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA. 8 Department of Pathology and Laboratory Medicine and Center for Computational Molecular Biology, Brown University, Providence, RI, USA. 9 Department of Geographical and Sustainability Sciences, University of Iowa, Iowa City, IA, USA. 10 These authors jointly supervised this work: Antoinette Tshefu, Jonathan B. Parr. e-mail: rachel.sendor@unc.edu; jonathan_parr@med.unc.edu Nature Communications | (2023)14:6618 1 1234567890():,; 1234567890():,;