Identification of micro-level malaria hotspots and its implications for achieving malaria elimination target in a remote district of India.

Challenges and strategies in malaria control: A case series from East Azerbaijan Province, 2023-2024.

The elimination of Plasmodium falciparum malaria in Eswatini remains elusive because of ongoing importation and sustained local transmission. The current status of antimalarial and diagnostic resistance in the country remains unknown. Genomic surveillance can complement routine surveillance by characterizing the prevalence and distribution of resistance markers and revealing granular patterns of transmission. Between March and December 2023, dried blood spots, demographic data, and travel history were collected from individuals who tested positive for malaria on a rapid diagnostic test (RDT) and from symptomatic RDT-negative individuals across Eswatini. Multiplexed amplicon deep sequencing was used to genotype 12 genes associated with antimalarial resistance, detect histidine-rich protein (hrp) 2/3 deletions, estimate genetic relatedness, and detect non-falciparum species. Data from 437 samples revealed significant clustering of parasites carrying resistance markers. A validated marker of artemisinin partial resistance, kelch13 P553L, was identified in a cluster of four infections (0.9% of all samples), likely linked to importation. The dihydropteroate synthase/dihydrofolate reductase sextuple mutant haplotype, associated with high-level sulfadoxine-pyrimethamine resistance, was found in 7.8% of the samples, also within clusters. The multidrug resistance 1 N86 genotype associated with reduced susceptibility to lumefantrine was nearly fixed. More than 60% of the infections were polyclonal, with higher complexity observed in imported cases. No hrp2/3 deletions were detected; most false-negative RDTs were attributed to subpatent parasitemia. Non-falciparum coinfections were rare (<1%). These findings shed light on the dynamics of resistance emergence in low-transmission settings. Integrating routine genomic surveillance into national malaria programs is essential for detecting, tracking, and responding to resistance threats in countries nearing elimination.

Each year, Plasmodium-infected mosquitoes cause approximately 250 million cases of malaria within 85 countries containing half of the world's population. Rapid diagnostic tests (RDTs) for blood-based biomarkers of the parasite can accelerate and expand access to diagnostic testing. Malaria RDTs have been endorsed by the WHO for first-line malaria testing, and next-generation assays are being developed to detect multiple Plasmodium antigens. If drug resistance continues to expand, then future RDTs to detect drug resistance will be beneficial. Because malaria RDTs will be an essential diagnostic tool for malaria elimination, particularly in high-burden malaria-endemic countries, we conducted a landscape review and analysis by gathering current information on the advancements in malaria RDTs that are either commercially available or in late-stage development. Overall, we identified 67 malaria RDTs from 27 different international manufacturers. Among these RDTs, most are simple to use on a whole blood specimen, provide test results within 30 minutes, and have reported high (≥98%) diagnostic sensitivity and specificity. Although many malaria RDTs have received WHO approval, clinical and implementation science studies will be needed to determine optimal and efficient delivery and care strategies for malaria RDTs in malaria-endemic settings.

Assessing the effect of parasite antigen genetic diversity on performance of Plasmodium vivax serological exposure markers for malaria: a multicentre observational diagnostic accuracy study.

Artemether-lumefantrine for the treatment of Plasmodium falciparum malaria in Laos: a therapeutic efficacy study coupled with genomic and in vitro phenotypic analyses.

Thailand has achieved significant progress in malaria elimination, with a reduction in annual parasite incidence from 0.53 to 0.22 per thousand in 2014 and 2024, respectively. Given the high diversity of Anopheles mosquito species, elimination efforts must be precisely targeted, taking into account the varied behaviors and vectorial capacities of different vector species. This study aims to systematically review and update the distribution, identification, bionomics, behavior, and a meta-analysis of nonhuman parasite infectivity among mosquitoes. A comprehensive literature search was conducted in PubMed, Scopus, EBSCOhost, and Google Scholar (2013-2025) to identify studies on Anopheles species diversity, distribution, and zoonotic malaria infection in mosquitoes. The meta-analysis followed Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines and was carried out using the metafor package in R. A total of 92 relevant papers were included from 811 accessed articles. Of these, most documented geographical distribution, followed by mosquito behaviors, molecular identification, and mosquito infectivity. The pooled mosquito infection prevalence for the present meta-analysis was 0.01 (95% confidence interval (CI) 0.00-0.03), indicating low nonhuman primate (NHP) malaria parasite infectivity. Using standard nested polymerase chain reaction (PCR), wild-caught Anopheles sawadwongporni, An. minimus, and An. dirus were incriminated as zoonotic malaria vectors, with a pooled infection prevalence of 0.2%. This review highlights a critical need for targeted, context-specific vector control interventions that address the unique opportunistic feeding and resting behaviors of Anopheles species complexes and the bionomics of local vector species, alongside the high mobility of high-risk populations. Research involving hosts and vectors across different micro-spatial and temporal scales, observations of human behavior, and monkey-human interactions is needed to improve understanding of zoonotic malaria transmission.

Malaria transmission-blocking vaccines (TBVs) targeting parasite transmission by mosquitoes represent a valuable public health tool for malaria control and elimination through herd immunity in the community. Pfs230, a surface protein expressed in gametocytes and gametes of Plasmodium falciparum, plays a critical role in gamete fertilization and further development within mosquitoes. Prior studies have advanced the N-terminal pro-domain (Pro) and domain 1 (D1) of Pfs230 as a putative TBV target. However, whether other Pfs230 domains function as TBV candidates needs further study. DNA vaccines encoding Pfs230 domains D7 to D10 (D7D10) were developed and evaluated in BALB/c mice. Antigen-specific antibody responses were assessed via enzyme-linked immunosorbent assay. Transmission-reducing activity (TRA) of antibodies induced by D7D10 DNA and protein vaccines were evaluated in standard membrane feeding assays (SMFAs) using in vitro-cultured P falciparum gametocytes. In addition, we investigated functional activity of antibodies induced by DNA vaccines encoding smaller subdomains D7 and D7D8. Potent D7D10-specific antibody responses were elicited in mice immunized with DNA vaccines. Anti-D7D10 IgGs purified from mice revealed strong dose-dependent TRA against P falciparum in SMFA, irrespective of the presence or absence of active complement. Western blot analysis using various Pfs230 fragments (D5-D12) suggested that antibodies elicited by D7D10 vaccines predominantly target D7. Furthermore, DNA vaccines encoding D7 and D7D8 similarly induced antibodies with strong TRA in SMFAs. Our studies identify Pfs230-D7D10, particularly D7, as a novel and promising P falciparum TBV candidate.

Malaria remains a global health challenge, imposing a substantial economic burden on health systems and society. Comprehensive assessments of this burden have been limited. This study aimed to provide a detailed estimate of the economic burden of malaria in the Brazilian Amazon. Public health expenditures were derived from a previous study. Direct, and indirect household costs and Health-Related Quality-of-Life (HRQoL) losses were estimated using a household survey conducted in nine municipalities in five states of the Brazilian Amazon. Expenditure data were collected from 1,131 individuals who experienced a malaria episode between January 2019 and May 2022. All costs were monetized and converted to 2024 purchasing power parity US dollars (PPP-USD). The total economic burden of malaria in the Amazon in 2019 was approximately US$181.9 million PPP-USD. The Unified Health System (SUS) bore the largest share (72.4%), which was allocated primarily to control and prevention activities. The states of Amazonas, Pará, and Roraima bore the highest total burden. Household burden was predominantly driven by indirect and mortality costs, and HRQoL losses. Major limitations of the study included convenience sampling and potential recall bias regarding expenditures; the latter was mitigated by surveying items and quantities rather than monetary values. This study offers a comprehensive and unparalleled assessment of the economic burden of malaria, providing a foundation for developing effective malaria control policies at the national and local levels in Brazil. Our findings underscore the critical role of the SUS in shielding families from direct medical costs, while revealing the substantial indirect costs, health-related quality-of-life losses, and mortality-related losses borne by households. These insights are essential for informing resource allocation and strengthening malaria control and elimination strategies in the Brazilian Amazon. The study provides a conceptual framework for estimating the economic cost of malaria that can be adapted to other malaria-endemic countries.

Plasmodium vivax is the main cause of malaria outside of sub-Saharan Africa, and in many settings it presents significant challenges to malaria elimination efforts. Despite some control successes in the Americas, regional annual case counts of malaria have increased by over 25% between 2014 and 2023, largely driven by P. vivax. Genomic surveillance can play a key role in understanding the extent to which disease persistence represents indigenous transmission as opposed to introduction of new strains through migration, and whether specific variants evade control measures. Efforts to make P. vivax genomic surveillance more cost-effective have led to the development of targeted sequencing-based methods, which strike a varying balance between assay sensitivity and breadth/informativeness. We introduce two new highly sensitive multiplexed amplicon sequencing panels for P. vivax: PvGTSeq and PvCRiSP. PvGTSeq requires selective whole-genome amplification (sWGA) and contains 249 amplicons-36 for antimalarial resistance and 213 for population structure-optimized for Latin America but applicable to all continents. PvCRiSP features four highly polymorphic amplicons that operate without sWGA and is designed to estimate complexity of infection (COI), identify instances of clonal transmission, and characterize recurrent episodes. Both panels use a single multiplex PCR with non-proprietary reagents, achieve ≥75% amplicon recovery at parasitemias as low as five parasites/μL, and PvCRiSP remains effective with low quality DNA. PvGTSeq showed high sequencing accuracy (error rate 3.85e-4% - 2.87e-3%), and both panels efficiently detected alleles from minority clones in simulated polyclonal infections. We validated both panels with samples from Colombia, Guyana, Honduras, Panama, and Venezuela, and performed in-silico assessments using data from 16 countries worldwide, confirming that these two panels have high power to discriminate samples and assign global geographic origin to imported cases. These panels will therefore be useful tools for P. vivax molecular surveillance in diverse geographic settings.

Following the elimination of human malaria, Malaysia has experienced an increasing burden of zoonotic Plasmodium knowlesi malaria, particularly in Sabah. Evidence on the cost-effectiveness of vector control interventions is needed to inform policy decisions and optimise resource allocation. This study assessed the cost-effectiveness of Outdoor Residual Spraying (ORS), Long-Lasting Insecticidal Nets (LLINs), and their combined use for controlling P. knowlesi malaria in Sabah, Malaysian Borneo. A cost-effectiveness analysis was conducted from the healthcare provider perspective over a one-year time horizon in three high-endemic districts: Keningau (LLINs), Ranau (ORS), and Kota Marudu (ORS + LLINs). Capital and recurrent components, were estimated using an ingredient-based costing approach. Effectiveness was measured as the number of P. knowlesi cases averted based on routine surveillance data. Incremental cost-effectiveness ratios (ICERs) were calculated relative to a no-intervention scenario, defined as health promotion activities without vector control. One-way sensitivity analyses assessed uncertainty in key parameters. All costs were reported in Malaysian Ringgit (MYR) and adjusted to 2025 values. Annual provider costs ranged from MYR 3912.92 (no intervention) to MYR 39,764.14 (ORS + LLINs). ORS was associated with the highest number of cases averted (n = 13) and the lowest ICER (MYR 2046 per case averted; approximately USD 470). LLINs and the combined strategy were associated with fewer cases averted and higher ICERs. ORS consistently demonstrated the most favourable cost-effectiveness profile across all sensitivity analyses. ORS demonstrated the most favourable cost-effectiveness profile under the observed programme conditions for controlling P. knowlesi malaria in Sabah. A stratified approach prioritising ORS, with targeted use of LLINs among high-risk populations, may represent an economically efficient strategy in resource-limited settings.

Mosquito-borne diseases continue to exact a heavy toll on human health, particularly in sub-Saharan Africa. Yet in many environments, fundamental aspects of mosquito behavior and population dynamics remain poorly characterized. Here, we employ a well-established method for directly measuring mosquito dispersal and estimating population size: mark-release-recapture (MRR). We focused on the key malaria vector Anopheles coluzzii and conducted experiments in São Tomé and Príncipe, an island nation in the Gulf of Guinea under consideration as a site for the first field trial using gene drive mosquitoes for malaria elimination. Understanding mosquito dispersal, population size, and responses to environmental factors is essential for planning such releases. To assess these parameters in An. coluzzii, a total of four MRR experiments were conducted across both São Tomé and Príncipe islands during both wet and dry seasons. Population size estimates were higher during the wet season in both study areas, but seasonal fluctuation was more pronounced in São Tomé. Seasonal patterns of mosquito dispersal differed by location, with greater dispersal in São Tomé during the wet season and in Príncipe during the dry season. Mosquito flight direction was biased toward broad-scale wind direction in São Tomé, but not in Príncipe. Together, these results enhance our understanding of An. coluzzii behavior in island ecosystems and support the design of effective vector control approaches in this biogeographical context.

Despite decades of control efforts. malaria burden in Tanzania remains high, with marked heterogeneity in transmission intensity across regions. Insecticide-treated bed nets (ITNs) are a core malaria intervention and are distributed through multiple channels in Tanzania to promote equitable access and use, yet disparities in ITNs ownership and use persist. This study evaluated socio-demographic predictors of ITNs ownership and use among rural communities from five regions with varying malaria endemicity. A community-based cross-sectional survey covering individuals aged ≥ 6months was conducted from July to August 2023 in 15 villages across five districts from five regions of Mainland Tanzania (Kagera, Kigoma, Njombe, Ruvuma, and Tanga). Data on demographics, malaria prevention practices, anthropometrics and socio-economic status (SES) were collected using structured questionnaires installed in tablets, run with Open Data Kit (ODK) software. Socio-demographic predictors of ITNs ownership and use were assessed using logistic regression analysis. The results were reported as crude (cOR) and adjusted odds ratios (aOR) with 95% confidence intervals (CI) and a p-value < 0.05 was considered statistically significant. Among the 10,228 enrolled participants, 7939 (77.6%) and 7899 (77.2%) reported owning and using ITNs, respectively. ITNs ownership and use varied significantly across districts (p < 0.001), with the highest rates observed in Nyasa (Ruvuma) and the lowest in Kyerwa (Kagera). Females had higher odds of both ITNs ownership and use than males (aOR = 1.27, 95% CI 1.12-1.45, p < 0.001 for both outcomes). Under-fives were more likely to own (aOR = 1.83, 95%CI 1.56-2.15, p < 0.001) and use ITNs (aOR = 2.26, 95%CI 1.62-3.15, p < 0.001) than adults. Participants from Nyasa (Ruvuma), Ludewa (Njombe), Muheza (Tanga) and Buhigwe (Kigoma) districts exhibited higher odds of ITNs ownership and use compared to those from Kyerwa (Kagera) (p < 0.001). Higher education attainment and household SES were independently associated with increased ITNs ownership and use (p < 0.001). Although ITNs ownership and use were relatively higher across the surveyed communities, coverage remained below the national target of 80% (projected for 2023). Higher ITNs ownership and use were reported among females, under-fives, participants with higher education and those from households with high SES. Disparities by sex, age groups, household SES and education status persist and should be explicitly addressed through ITNs distribution strategies to enable equitable access and use of ITNs across all population groups to expedite progress toward malaria elimination in Tanzania.

Malaria elimination remains a global public health priority, particularly in low- and middle-income countries. India has set a target of eliminating malaria by 2030, making it essential to understand gaps in healthcare access, utilization, and preventive practices in endemic areas to guide focused interventions to achieve national and global malaria elimination targets. A subnational, cross-sectional study surveying 37,849 households in ten malaria-endemic states in India, which covered 177,644 individuals, was carried out to estimate the health service and long-lasting insecticidal net (LLIN) utilization for febrile illness and malaria. The study found that the accessibility of a government healthcare facility was within 2.5km (interquartile range [IQR] 1-5km), with 57.4% of febrile individuals visiting a government healthcare center or approaching a government-designated healthcare provider for fever within 24-48h. Adequate knowledge and behavior regarding prevention were observed in 89.2% households, while 73.6% households reported using LLINs regularly. Lack of formal education and belonging to the tribal population were found to be high risk factors for malaria. Healthcare-seeking behavior played a crucial role in malaria burden. This study highlights the gap between accessibility of health facility and its utilization, as well as LLIN ownership and its regular utilization.

Malaria remains a leading global public health concern, disproportionately affecting populations in low-resource settings. Uganda continues to contribute substantially to the global malaria burden, yet exposure to malaria-related health messages remains limited. In recent years, diverse media platforms have been adopted to disseminate prevention messages. This study assessed the influence of cumulative exposure to malaria message media channels on knowledge of malaria prevention and its associated factors among women of reproductive age in Uganda. We conducted a secondary analysis of the 2018-2019 Uganda Malaria Indicator Survey (UMIS), comprising 7124 women aged 15-49years selected using a two-stage cluster and stratified sampling design. Knowledge of four prevention methodsrecommended by World Health Organization (WHO), bed nets, insecticide-treated nets (ITNs), preventive medicine, and indoor residual spraying (IRS) was assessed. Associations between cumulative media exposure and malaria knowledge were examined using t-tests, margins analysis, and mixed-effects negative binomial regression models. Knowledge of malaria prevention was unevenly distributed: 76.5% of women reported awareness of bed nets, compared with only 9.1% for ITNs, 6.2% for preventive medicine, and 4.3% for IRS. Cumulative exposure to media channels was significantly associated with knowledge of ITNs (p < 0.001), preventive medicine (p = 0.002), and IRS (p < 0.001), but not bed nets. Education, age, wealth, residence, and region were significant determinants of exposure to media channels. Women with secondary or higher education were nearly twice as likely to report exposure to multiple channels compared to uneducated women (Incidence Rate Ratio,IRR = 1.86; 95% Confidence Interval,CI 1.59-2.17). Similarly, women aged ≥ 40years were 69% more likely (IRR = 1.69; 95% CI 1.35-2.11) to report exposure to multiple channels than those under 20years. In contrast, rural and refugee women reported significantly lower exposure relative to urban residents (IRR = 0.77 and 0.28, respectively). Cumulative multi-channel exposure to malaria prevention messages significantly improves women's knowledge of ITNs, preventive medicine, and IRS. However, structural inequities in education, wealth, and place of residence limit access to diverse channels. Integrated, context-specific, and equity-focused communication strategies are essential to broaden awareness beyond bed nets and to accelerate progress toward malaria control and elimination in Uganda.

Sudan is one of the few African countries reporting increasing Plasmodium vivax infections despite the high prevalence of the Duffy-negative phenotype, historically considered protective against this parasite. Emerging molecular evidence challenges this paradigm. The objective of this systematic review and meta-analysis was to synthesize evidence on the prevalence and geographic distribution of P. vivax infection, Duffy antigen polymorphisms, parasite genetic adaptations, and diagnostic limitations in Sudan. This review followed PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines and was registered in PROSPERO (CRD42025127714). The reports of P. vivax infection in Sudan focusing on host Duffy status and the parasite genetic variability from 2005 to 2025 were identified through systematic searches of PubMed, Web of Science, Scopus, and EMBASE with predefined Boolean search operators. To determine the risk of bias, the Joanna Briggs Institute prevalence study checklist was employed. To obtain a pooled prevalence estimate in Duffy-negative individuals, a random-effects meta-analysis (DerSimonian-Laird estimator) was conducted. Heterogeneity was assessed using Cochran's Q test and the I2 statistic, while publication bias was evaluated using funnel plots and Egger's regression test. A total of sixteen studies conducted between 2005 and 2025, including 5,753 participants from various regions of Sudan, were included. P. vivax infection was reported in both Duffy-positive and Duffy-negative individuals. Meta-analysis of five studies reporting host Duffy antigen status showed a pooled prevalence of P. vivax infection of 11.7% (95% CI 7.2-17.3%) among Duffy-negative individuals. The studies showed moderate heterogeneity (I2 = 56%). Substantial genetic diversity was observed in the P. vivax Duffy Binding Protein (PvDBP), including multiple haplotypes and both Malagasy-type and Cambodian-type gene duplications. Most studies were assessed as having low to moderate risk of bias, and funnel plot inspection did not suggest substantial publication bias, although interpretation is limited by the small number of studies. This systematic review and meta-analysis provide evidence that P. vivax infection occurs among Duffy-negative individuals in Sudan, challenging the long-standing assumption of complete Duffy-mediated protection. The observed parasite genetic diversity highlights adaptive mechanisms that may facilitate infection in Duffy-negative hosts. Strengthening molecular surveillance and integrating host-parasite genomic data into national malaria control programs will be critical to inform malaria elimination strategies and vaccine development in Africa.

Abstract Identifying and characterizing Anopheles mosquitoes through spot surveys is critical for malaria control, especially in endemic and receptive areas. In the Special Region of Yogyakarta, Indonesia, malaria cases have been reported in Kulonprogo, with the latest 10 cases in 2023. This study aimed to identify Anopheles species and their resting behavior as risk factors for local malaria transmission. A cross-sectional entomological survey was conducted in Samigaluh, Kulonprogo. Mosquitoes were collected using human landing catches (indoor and outdoor) and resting collections in animal shelters from 6:00 PM to 6:00 AM. Specimens were identified by species, vector density, and parity status. Five Anopheles species were detected, predominantly resting in animal shelters: An. vagus (71.9%), An. annularis (21.9%), An. aconitus (3.95%), An. kochi (1.69%), and An. barbirostris (0.56%). All biting activity occurred outdoors between 6:00 PM and 3:00 AM, with species-specific activity peaks. An. vagus had the highest man biting rate (0.88 bites/person/hour) and was active early in the evening. The overall parity rate was 85.71%, indicating a mature mosquito population with potential for malaria transmission. These findings highlight An. vagus as the dominant species in the area, exhibiting exophagic and zoophilic behavior, and suggest that sustained vector surveillance is essential to support malaria elimination efforts in receptive regions.

Malaria transmission at the human-wildlife interface remains poorly characterized in the Amazon. We conducted a molecular survey of Plasmodium spp. in an Indigenous community (n = 141) and sympatric nonhuman primates (NHPs) (n = 341; 10 species) in the Peruvian Amazon during 2007-2020. By using nested or quantitative PCR (targeting cytb, cox3, and 18S rRNA genes) and sequencing, we estimated prevalence, parasite load, and genetic similarity. We detected Plasmodium in 43.3% of humans and 51.9% of NHPs. P. vivax/simium predominated in humans (42.1%), whereas P. brasilianum/malariae predominated in NHPs (24.6%). P. falciparum was rare in both hosts. Children <8 years of age showed higher parasite load than older persons. Bayesian phylogenies revealed >99.9% identity among human and NHP lineages, supporting shared Plasmodium lineages. NHP lineages showed low interannual variation. One third of human infections were asymptomatic. Our findings reveal hidden reservoirs and support integrating wildlife surveillance into Amazon malaria elimination strategies.

Early and accurate detection of malaria is essential for appropriate case management, minimizing unnecessary treatment, and supporting surveillance efforts crucial for disease control and elimination. Microscopy has long been the reference method for malaria diagnosis. Rapid diagnostic tests (RDTs) have revolutionised point-of-care diagnosis of malaria, with billions of units distributed globally over the past decade. However, ensuring their reliability necessitates rigorous quality assurance measures. Inaccurate test performance can lead to misdiagnosis, inappropriate treatment, increased mortality, and the potential emergence of drug-resistant strains that can lead to delay in achieving the target of malaria elimination. Strengthening quality assurance protocols is crucial to maintain the accuracy of malaria diagnostics, enhancing disease surveillance, and bolstering global malaria elimination initiatives. This paper focuses on the role of quality assurance in optimising the performance and reliability of malaria RDTs. By implementing enhanced quality assurance frameworks, diagnostic accuracy can be safeguarded, toward malaria eradication.

Malaria is one of the major global public health issues. An estimated 282 million malaria cases occurred worldwide in 2024, and the overall prevention and control progress has stagnated or even reversed in some regions. Mass drug administration (MDA), as a potential strategy to accelerate malaria elimination, has regained attention. This paper reviews the evidence base, controversial focuses, and application strategies of MDA in malaria prevention and control. It aims to promote its scientific application in the elimination phase. MDA plays an important role in malaria prevention and control. However, this strategy is accompanied by core limitations such as long-term drug resistance risks, insufficient implementation sustainability, and a high failure rate of regional adaptation. It also faces challenges from multiple common malaria species, as well as the newly discovered Plasmodium knowlesi. We therefore propose an “MDA+” collaborative strategy integrating vaccines, digital monitoring, and cross-border cooperation, so as to optimize resource allocation, achieve full coverage control over various malaria parasites, and advance the global malaria elimination process.