Eric Rogier

Background: Nigeria consistently shows the greatest population burden of Plasmodium falciparum (Pf) malaria. While rapid diagnostic tests and microscopy offer valuable information for clinical decisions and prevalence estimates, serological tools provide more nuanced information on exposure history, enhancing the ability to assess and track malaria burden at a population level. Methods: Blood samples (n = 89 438) from a nationwide 2018 human immunodeficiency virus household survey in Nigeria were assayed for active/recent infection by testing for the Pf HRP2 and LDH antigens, and for Pf exposure by immunoglobulin G carriage against a panel of antigens. Risk factors for Pf infection and exposure were assessed using logistic regression and geospatial analyses. Results: By blood antigenemia, current/recent Pf prevalence was 38.3% (95% CI: 37.2%–39.5%), and seroprevalence among the panel of 5 Pf antigens ranged from 48% to 87%. In multivariate regression models, participants age 5–14 years had the highest odds of active infection versus children <5 years. Compared with the lowest wealth category, the wealthiest category showed a 5-fold reduction in odds of active infection, and urban living was associated with a 30% reduction. In estimating a longer window of Pf exposure, serology recapitulated these associations with the highest wealth category showing a 10-fold reduction, and urban living a 60% reduction, in exposure. Conclusions: Geospatial findings underscored Nigeria's hyperendemic but heterogeneous malaria epidemiology with infection and exposure are not unequally distributed among the populace. Serological data can provide greater insights into malaria risk factor analyses and augment examination of spatial heterogeneities in transmission.

Background: The Zanzibar archipelago has been a pre-elimination region for malaria thanks to rigorous control interventions, but recent surges in malaria cases have been observed. The contribution of non-falciparum species to the current malaria situation is unknown. Method: This study investigates the epidemiology of falciparum and non-falciparum malaria species in Zanzibar. Leveraging plasma extracted from dried blood spots (DBS) collected during reactive case detection (RCD) activities across Unguja island from May 2022 to January 2023, we measured immunoglobulin G (IgG) responses to Plasmodium MSP1-19kD antigens using a multiplex bead-based immunoassay. Additionally, active infections were detected using species-specific real-time PCR. Results: Out of 1618 participants surveyed in 35 RCDs, 35.3% had exposure to any malaria species, with Plasmodium falciparum being the most common (29.8%). Seroprevalences for non-falciparum species were lower: Plasmodium ovale (5.8%), Plasmodium malariae (5.9%), and Plasmodium vivax (5.9%). Active infections were detected in 6.0% of participants, predominantly P. falciparum (4.6%). Travel to mainland Tanzania was a dominant risk factor for seropositivity for all 4 malaria species. Other factors associated with Pf seropositivity (high-risk occupations and female status) were not associated with seropositivity for non-falciparum species. The geographic distribution of non-falciparum exposure differed compared to falciparum, with relatively higher seroprevalences in rural districts, especially Kazkazini A in northern Unguja. Conclusions: This study suggests a significant contribution of non-falciparum species to the local epidemiology in Zanzibar. Current control and elimination efforts, focused on P. falciparum, may not adequately address exposure to non-falciparum species.

: Hispaniola, which is shared by Haiti and the Dominican Republic, remains the last island in the Caribbean that is still endemic for malaria, with Haiti bearing the highest caseload. Few studies have examined the ecology of malaria vectors in Haiti. Five species of Anopheles have been described on the island, but the exophilic Anopheles albimanus (An. albimanus) is considered the primary vector of malaria in Haiti. Households recruited for a case–control study profiling risk factors for symptomatic Plasmodium falciparum (P. falciparum) infections were approached to participate in an entomological study. The goal was to determine the bionomics of anopheline mosquitoes around the 32 participating households across varying malaria transmission settings. We assessed the characteristics of the Anopheles population using ultraviolet-light traps and larval surveys. Anopheles albimanus was the most abundant mosquito species identified in the Grand’Anse. Its abundance was higher in outdoor traps than in indoor traps and in areas with relatively high positivity based on rapid diagnostic test results. A greater proportion of blood-fed mosquitoes were found in higher transmission areas. Anopheles albimanus samples were found to be infected with both P. falciparum and Plasmodium vivax sporozoites. As Haiti aims for the elimination of malaria, disrupting localized residual malaria transmission will increasingly rely on focal vector control strategies.

: Many SARS-CoV-2 infections are asymptomatic, thus reported cases underestimate actual cases. To improve estimates, we conducted surveillance for SARS-CoV-2 seroprevalence among pregnant women attending their first antenatal care visit (ANC1) from June 2021 through May 2022. We administered a questionnaire to collect demographic, risk factors, and COVID-19 vaccine status information and tested dried blood spots for SARS-CoV-2 antibodies. Although <1% of ANC1 participants reported having had COVID-19, monthly SARS-CoV-2 seroprevalence increased from 15.4% (95% CI: 10.5–21.5) in June 2021 to 65.5% (95% CI: 55.5–73.7) in May 2022. Although COVID-19 vaccination was available in March 2021, uptake remained low, reaching a maximum of 9.5% (95% CI: 5.7–14.8) in May 2022. Results of ANC1 serosurveillance provided prevalence estimates helpful in understanding this population case burden that was available through self-report and national case reports. To improve vaccine uptake, efforts to address fears and misconceptions regarding COVID-19 vaccines are needed.

Background: We assessed associations between binding antibody (bAb) concentration <5 days from symptom onset and testing positive for COVID-19 among patients in a test-negative study. Methods: From October 2021 to June 2022, study sites in 7 states enrolled patients aged ≥6 months presenting with acute respiratory illness. Respiratory specimens were tested for SARS-CoV-2. In blood specimens, we measured concentrations of anti-SARS-CoV-2 antibodies against the spike protein receptor binding domain (RBD) and nucleocapsid antigens from the ancestral strain in standardized bAb units (BAU). Percentage change in odds of COVID-19 by increasing anti-RBD bAb was estimated via logistic regression as (1 – adjusted odds ratio of COVID-19) × 100, adjusting for COVID-19 mRNA vaccine doses, age, site, and high-risk exposure. Results: Out of 2018 symptomatic patients, 662 (33%) tested positive for acute SARS-CoV-2 infection. Geometric mean RBD bAb levels were lower among COVID-19 cases than SARS-CoV-2 test-negative controls during the Delta-predominant period (112 vs 498 BAU/mL) and Omicron-predominant period (823 vs 1189 BAU/mL). Acute-phase ancestral spike RBD bAb levels associated with 50% lower odds of COVID-19 were 1968 BAU/mL against Delta and 3375 BAU/mL against Omicron; thresholds may differ in other laboratories. Conclusions: During acute illness, antibody concentrations against ancestral spike RBD were associated with protection against COVID-19.

Background: Due to low numbers of active infections and persons presenting to health facilities for malaria treatment, case-based surveillance is inefficient for understanding the remaining disease burden in low malaria transmission settings. Serological data through the detection of IgG antibodies from previous malaria parasite exposure can fill this gap by providing a nuanced picture of where sustained transmission remains. Study enrollment at sites of gathering provides a potential approach to spatially estimate malaria exposure and could preclude the need for more intensive community-based sampling. Methods: This study compared spatial estimates of malaria exposure from cross-sectional school- and community-based sampling in Haiti. A total of 52,405 blood samples were collected from 2012 to 2017. Multiplex bead assays (MBAs) tested IgG against P. falciparum liver stage antigen-1 (LSA-1), apical membrane antigen 1 (AMA1), and merozoite surface protein 1 (MSP1). Predictive geospatial models of seropositivity adjusted for environmental covariates, and results were compared using correlations by coordinate points and communes across Haiti. Results: Consistent directional associations were observed between seroprevalence and environmental covariates for elevation (negative), air temperature (negative), and travel time to urban centers (positive). Spearman’s rank correlation for predicted seroprevalence at coordinate points was lowest for LSA-1 (ρ = 0.10, 95% CI: 0.09–0.11), but improved for AMA1 (ρ = 0.36, 95% CI: 0.35–0.37) and MSP1 (ρ = 0.48, 95% CI: 0.47–0.49). Conclusions: In settings approaching P. falciparum elimination, case-based prevalence data does not provide a resolution of ongoing malaria transmission in the population. Immunogenic antigen targets (e.g., AMA1, MSP1) that give higher population rates of seropositivity provide moderate correlation to gold standard community sampling designs and are a feasible approach to discern foci of residual P. falciparum transmission in an area.

To describe humoral immune responses to symptomatic SARS-CoV-2 infection, we assessed immunoglobulin G binding antibody levels using a commercial multiplex bead assay against SARS-CoV-2 ancestral spike protein receptor binding domain (RBD) and nucleocapsid protein (N). We measured binding antibody units per mL (BAU/mL) during acute illness within 5 days of illness onset and during convalescence in 105 ambulatory patients with laboratory-confirmed SARS-CoV-2 infection with Omicron variant viruses. Comparing acute- to convalescent phase antibody concentrations, geometric mean anti-N antibody concentrations increased 47-fold from 5.5 to 259 BAU/mL. Anti-RBD antibody concentrations increased 2.5-fold from 1258 to 3189 BAU/mL.

Background: Plasmodium falciparum malaria is a leading cause of child mortality in Nigeria. Neonates are born with maternal antibodies from placental transfer which may protect against malaria infection in the first months of life. The IgG dynamics of the transition from passively transferred antimalarial antibodies to actively acquired IgG from natural exposure have not been well elucidated.Methods: Blood samples collected during a 2018 Nigeria nationwide HIV/AIDS household survey were available for 9,443 children under 5 years of age, with a subset of infants under 2 months of age having maternal samples available (n=41). Samples were assayed for the P. falciparum HRP2 antigen and anti-malarial IgG antibodies. LOESS regression examined the dynamics in IgG response in the first 5 years of life. Correlation with maternal IgG levels was assessed for mother/child pairs.Results: Consistent decreases were observed in median IgG levels against all Plasmodium spp. antigen targets for the first months of life. At a population level, P. falciparum apical membrane antigen-1 (AMA1) and merozoite surface protein-1 19kD (PfMSP1) IgG decreased during the first 12 months of life before reaching a nadir, whereas IgGs to other targets only declined for the first 4 months of life. Seropositivity showed a similar decline with the lowest seropositivity against AMA1 and PfMSP1 at 10-12 months, though remaining above 50% during the first 2 years of life in higher transmission areas. No protective association was observed between IgG positivity and P. falciparum infection in infants. Maternal antibody levels showed a strong positive correlation with infant antibody levels for all P. falciparum antigens from birth to 2 months of age, but this correlation was lost by 6 months of age.Discussion: Maternally transferred anti-malarial IgG antibodies rapidly decline during the first 6 months of life, with variations among specific antigens and malaria transmission intensity. From 3-23 months of age, there was a wide range in IgG levels for the blood-stage antigens indicating high individual variation in antibody production as children are infected with malaria. Non-falciparum species-specific antigens showed similar patterns in waning immunity and correlation with paired mother’s IgG levels compared to P. falciparum antigens.

: Targeting malaria interventions in elimination settings where transmission is heterogeneous is essential to ensure the efficient use of resources. Identifying the most important risk factors among persons experiencing a range of exposure can facilitate such targeting. A cross-sectional household survey was conducted in Artibonite, Haiti, to identify and characterize spatial clustering of malaria infections. Household members (N = 21,813) from 6,962 households were surveyed and tested for malaria. An infection was defined as testing positive for Plasmodium falciparum by either a conventional or novel highly sensitive rapid diagnostic test. Seropositivity to the early transcribed membrane protein 5 antigen 1 represented recent exposure to P. falciparum. Clusters were identified using SaTScan. Associations among individual, household, and environmental risk factors for malaria, recent exposure, and living in spatial clusters of these outcomes were evaluated. Malaria infection was detected in 161 individuals (median age: 15 years). Weighted malaria prevalence was low (0.56%; 95% CI: 0.45–0.70%). Serological evidence of recent exposure was detected in 1,134 individuals. Bed net use, household wealth, and elevation were protective, whereas being febrile, over age 5 years, and living in either households with rudimentary wall material or farther from the road increased the odds of malaria. Two predominant overlapping spatial clusters of infection and recent exposure were identified. Individual, household, and environmental risk factors are associated with the odds of individual risk and recent exposure in Artibonite; spatial clusters are primarily associated with household-level risk factors. Findings from serology testing can further strengthen the targeting of interventions.

Mozambique is one of the four African countries which account for over half of all malaria deaths worldwide, yet little is known about the parasite genetic structure in that country. We performed P. falciparum amplicon and whole genome sequencing on 2251 malaria-infected blood samples collected in 2015 and 2018 in seven provinces of Mozambique to genotype antimalarial resistance markers and interrogate parasite population structure using genome-wide microhaplotyes. Here we show that the only resistance-associated markers observed at frequencies above 5% were pfmdr1-184F (59%), pfdhfr-51I/59 R/108 N (99%) and pfdhps-437G/540E (89%). The frequency of pfdhfr/pfdhps quintuple mutants associated with sulfadoxine-pyrimethamine resistance increased from 80% in 2015 to 89% in 2018 (p < 0.001), with a lower expected heterozygosity and higher relatedness of microhaplotypes surrounding pfdhps mutants than wild-type parasites suggestive of recent selection. pfdhfr/pfdhps quintuple mutants also increased from 72% in the north to 95% in the south (2018; p < 0.001). This resistance gradient was accompanied by a concentration of mutations at pfdhps-436 (17%) in the north, a south-to-north increase in the genetic complexity of P. falciparum infections (p = 0.001) and a microhaplotype signature of regional differentiation. The parasite population structure identified here offers insights to guide antimalarial interventions and epidemiological surveys.