Sustaining the roles of community health workers (CHWs) and community-based malaria services along the Thailand-Myanmar border is critical to maintain access to malaria prevention and care among mobile and migrant populations. As countries in the Greater Mekong Subregion (GMS) advance towards malaria elimination, there is an ideal policy window to integrate CHWs into the primary healthcare system and to identify innovative pathways for sustaining community-based malaria services. Participatory stakeholder engagement activities were conducted to document practical evidence and case studies of malaria service integration and sustained community-based malaria care in Kanchanaburi province, Thailand, along the border with Myanmar. Two multi-stakeholder workshops were conducted with 49 participants representing national, provincial, and community-level malaria actors. The activities were designed to identify gaps in access to malaria services among at-risk populations and in implementation of surveillance and control activities, as well as to inform changes that align with the health and wellbeing needs in border communities. Enabling factors and potential entry points for sustaining CHWs were identified. Stakeholders highlighted three core requirements for sustaining community-based malaria services: (1) comprehensive training with appropriate incentives for CHWs, (2) active community participation in malaria prevention and care, and (3) reliable policy and financial support. In high burden areas, immediate priorities included scaling up training for migrant health workers, while longer-term goals emphasized cross-border collaboration and integration of malaria surveillance into local health systems. The case studies illustrated diverse approaches to integrate community-based malaria care through financing mechanisms, harmonized training, public–private partnerships, and policy advocacy, each offering potential entry points for embedding malaria care within broader community health services. Findings from the Thailand-Myanmar border demonstrate that sustaining CHWs requires coordinated investment in skills, incentives, and system-level support, combined with approaches that reflect local priorities. By engaging stakeholders and documenting integration pathways, this study provides actionable strategies for programme implementers and partners to strengthen community-based malaria care, safeguard access for vulnerable populations, and co-create sustainable models aligned with broader health system strengthening. This collaborative approach offers valuable lessons for informing national strategies in the GMS ensuring they are well-suited to the communities they serve.
Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common enzymatic disorder with major implications for malaria control. Safe administration of radical cure antimalarials such as primaquine and tafenoquine requires reliable G6PD testing to prevent drug-induced hemolysis. This study aimed to evaluate the analytical performance of the Mindray BS-240 Analyzer using the new Twin-Test quantitative G6PD method and assess its suitability for routine diagnostics in malaria-endemic, resource-limited settings. A comparative analytical study was conducted at the Aga Khan University Hospital, Karachi, using archived EDTA-anticoagulated blood samples previously analyzed by the Trinity Biotech G6PD spectrophotometric assay. Samples were re-evaluated using the automated UV enzymatic method on the Mindray BS-240 Analyzer. Analytical performance was assessed through precision, trueness, linearity, method comparison, and carryover studies. Statistical analyses included coefficients of variation (CV), recovery rates, Deming regression, and Bland–Altman analysis. The Mindray BS-240 demonstrated high precision with CVs below 5
Genomic data are increasingly leveraged to enable malaria genomic surveillance, with many teams now generating Plasmodium falciparum sequence data for routine analysis. However, downstream analysis remains dependent on ad-hoc scripts and local infrastructure. This reliance limits reproducibility, and scalability, while obstructing the deployment of standardized pipelines across diverse high-performance computing (HPC) and cloud environments. This review describes the main workflow management systems and bioinformatics tools that are currently used for malaria genomic surveillance, with potential applicability to P. falciparum. We focus on the four most prominent workflow management systems (Nextflow, Snakemake, WDL/Cromwell, and Galaxy) and the core components of established pipelines in the field. Therefore, we summarize which tools are most widely used and how they are assembled into end-to-end workflows for surveillance. We organize these tools and pipelines around eight standardized PlasmoGenEPi use cases and propose a four-layer framework for pipeline design (data, pipeline structure, software environment, execution platform). Building on this, we introduce a pragmatic decision matrix that links sample volumes, infrastructure profiles and surveillance objectives to concrete choices of sequencing strategy and workflow system. We then discuss practical challenges that directly affect implementation in endemic settings, including heterogeneous computing infrastructure, long-term pipelines maintenance, staff turnover, and the systemic vulnerability of shared infrastructure, notably exposed by the recent VEuPathDB funding crisis. Finally, we highlight emerging directions, including community efforts such as PlasmoGenEPi, the increasing use of targeted Oxford Nanopore amplicon sequencing, and the move towards more standardized, portable and well-documented workflows that can be adapted to other pathogen surveillance systems.
Antenatal clinics serve as a key platform for pregnant women to access screening tests and preventive measures to reduce adverse maternal and pregnancy outcomes. In malaria-endemic settings, these include distribution of insecticide-treated bed nets, chemoprevention, and timely malaria diagnosis and treatment. However, the impact of these interventions has been hampered by suboptimal antenatal care (ANC) utilization. A systematic review of quantitative, qualitative, and mixed-methods studies from malaria-endemic countries, published between 1 January 2010 and 25 November 2024, was conducted to explore predictors of suboptimal ANC utilization. Suboptimal ANC was defined as no ANC utilization; late ANC initiation; and/or infrequent ANC utilization (<4 or <8 clinic contacts). Assessments for eligibility, quality, and data extraction were undertaken independently by two authors. Data were summarized and synthesized, and determinants of ANC utilization organized according to four levels of the Socio-Ecological Model: (i) intrapersonal; (ii) interpersonal; (iii) institutional and policy; and (iv) community. Random effects meta-analysis was conducted for homogenous quantitative data. A total of 70 quantitative, 45 qualitative, and 14 mixed-methods studies were included. Limited knowledge of ANC and pregnancy, low maternal educational status, nulli- and grand multi-parity, young maternal age, and poverty were critical intrapersonal factors related to suboptimal ANC utilization. Interpersonal influences from partners and other family members also shaped ANC utilization. Poor experiences of healthcare services and limited accessibility to clinics further limited ANC utilization. A range of institutional policies on mandatory partner attendance impacted on ANC utilization alongside socio-cultural norms. Several determinants of suboptimal ANC utilization intersected across Socio-Ecological Model levels. In malaria-endemic countries, a range of intertwined factors influence women’s ability to utilize ANC, where young, single and impoverished women are most vulnerable to suboptimal ANC utilization. Qualitative research critically complements observational data to identify drivers of poor ANC utilization that require multi-stakeholder, intersectoral collaboration to mitigate. PROSPERO registration number CRD42023384267.
Malaria transmission is influenced by climatic and environmental conditions, but the relative contributions of large-scale climate variability and local environmental factors remain unclear in southeastern Iran. We assessed associations between ENSO, remotely sensed environmental indicators, monsoon precipitation, and malaria incidence during 2001–2023. Monthly malaria surveillance data from eight counties in southeastern Iran (2001–2023) were analyzed using a retrospective ecological longitudinal design. Predictors included Niño 3.4 ENSO, NDVI, NDWI, and LST, with candidate lags evaluated based on model fit, estimated effects, and epidemiological plausibility. The primary analysis used a negative binomial mixed-effects model (NB-GLMM) with a log link, population offset, calendar-month adjustment, a four-degree-of-freedom nonlinear temporal trend, and county-level random intercepts. A quadratic term for six-month lagged LST was included to assess nonlinearity. County-specific models and random-slope analyses were used to assess spatial heterogeneity, while monsoon precipitation was analyzed separately using cumulative June–September rainfall in Chabahar, Saravan, and Sarbaz. In the final province-wide model, six-month lagged LST showed a significant nonlinear, U-shaped association with malaria incidence, with the estimated minimum occurring at approximately 0.32 standard deviations above the mean LST. ENSO (IRR = 1.01, 95
Ethiopia implemented the "1–3–7" surveillance and response approach for malaria elimination in selected districts. Evaluating its progress is crucial to ensure alignment with the national malaria elimination plan. This study assesses the approach's implementation and investigates challenges faced during its initial phase in the Farta district, northwest Ethiopia. A concurrent mixed-methods design was employed, combining a review of malaria elimination reports from April 2nd to December 24th, 2023, with interviews of 22 key stakeholders. Data were extracted from the district’s malaria surveillance database, and qualitative data were collected to explore the experiences and challenges of local health staff. The CDC’s guidelines for evaluating surveillance systems were used to assess the approach’s attributes. A total of 199 malaria cases were reported, with 85.4
Insecticide-treated nets (ITNs) remain one of the most cost-effective interventions for malaria prevention. However, ownership and consistent use remain suboptimal in many malaria-endemic settings. This study evaluated the determinants of ITN ownership and utilization among pregnant women attending antenatal clinics in selected hospitals in Enugu State, Nigeria. A cross-sectional study was conducted among 219 pregnant women attending antenatal care. Data were collected using a pretested structured questionnaire, and malaria infection was diagnosed by microscopy. Associations between variables were assessed using odds ratio (OR) with 95
Anopheles (An.) stephensi (Liston, 1901) was first detected on the African continent in 2012 in Djibouti. Its arrival is implicated in sharp increases in malaria cases in Djibouti: from 2012 to 2021 there was a > 2500-fold rise in case numbers, reaching > 70,000 cases in 2021. Unlike other malaria vectors, An. stephensi is well-adapted to colonising urban environments. Existing control tools provide inadequate protection, and the mosquito is resistant to commonly used insecticides. An. stephensi has now spread to other countries in Eastern and Western Africa, posing a new urban malaria threat across the continent. To support the development of future An. stephensi management strategies, and to inform deployments of a new genetically engineered self-limiting An. stephensi, we conducted studies to characterise population dynamics and insecticide resistance profiles of An. stephensi in Djibouti. From December 2021 to January 2023, monitoring of adult An. stephensi was conducted using odour-baited BG-Pro traps in urban and rural/semi-rural communities in Djibouti to characterise the vector’s distribution and seasonal dynamics. Data were also collected on the biotypes using OBP1 sequencing and the prevalence of known insecticide resistance-associated markers using PCR-based methods. An. stephensi abundance increased from January to April 2022, after which numbers declined during the hot summer season, followed by a further population increase from August to October 2022, before falling to low levels until the end of the monitoring period. All three biotypes of An. stephensi were detected, with the majority being type and intermediate, both of which are known to be competent malaria vectors. The target site mutation L1014F, associated with pyrethroid insecticide resistance, was found in 60
Long-Lasting Insecticidal Nets (LLINs) are a key tool for control and elimination of malaria, by providing protection from the bite of infected mosquitoes. To support its malaria elimination goal by 2030, Gujarat distributed LLINs across 33 districts in 2019. However, sustained effectiveness depends on consistent use and proper maintenance. A household-based cross-sectional survey was conducted in the Anand, Kutch, Panchmahal, and Surendranagar districts of the state of Gujarat in India. A Standard WHO questionnaire was used to assess LLIN coverage, usage, and maintenance. A total of 4719 households were surveyed. Net ownership, usage patterns, physical integrity (Hole size, washing and dry pattern), and maintenance practices were evaluated. LLIN ownership was high (93.8
South Sudan has one of the highest malaria burdens per capita in Africa, and artesunate-amodiaquine (ASAQ) and artemether-lumefantrine (AL) are first- and second-line treatments for uncomplicated Plasmodium falciparum malaria. However, despite the emergence of Artemisinin partial resistance (ART-R) and AL failure at some sites in the region, no therapeutic efficacy study (TES) has been conducted since the introduction of ASAQ and AL two decades ago. Our TES aimed to assess the efficacy of ASAQ and AL across four sites in South Sudan. This study was conducted in Juba, Bor, Wau, and Yambio. Children aged 6 months to 59 months with microscopically confirmed uncomplicated P. falciparum monoinfection were assigned to ASAQ or AL and followed for 28 days to assess treatment response. The primary endpoint was the Kaplan–Meier PCR-corrected adequate clinical and parasitological response (ACPR) on Day 28. Day 3 parasite positivity was used as a measure of early treatment response. We enrolled 390 children across all arms; 10.3
Malaria remains a significant public health concern in Thailand, particularly in Tak Province along the Thailand–Myanmar border. Despite ongoing elimination efforts, persistent transmission in endemic communities highlights the need to better understand prevention behaviors and associated barriers. This study aimed to identify factors influencing malaria prevention behaviors among residents in endemic border areas using the Health Belief Model framework. A convergent mixed-method study was conducted in four malaria-endemic villages in Khane Chue Subdistrict, Mae Ramat District, Tak Province, Thailand. Quantitative data were collected from 352 participants through face-to-face interviews using structured questionnaires. Qualitative data were obtained from in-depth interviews with 9 villagers. Quantitative data were analyzed using descriptive statistics, chi-square tests, Fisher’s exact test, and multivariable logistic regression. Qualitative data were analyzed using inductive thematic analysis. Overall, 77.3
Kenya has made notable progress in reducing malaria prevalence and improving child survival, yet the lake-endemic region continues to face high transmission and substantial malaria-related illness and death. To strengthen prevention, the RTS,S/AS01 malaria vaccine was introduced in 2019 through Kenya’s Expanded Programme on Immunization (EPI), leveraging a robust platform with decades of experience in rolling out new vaccines. This situational analysis provides a comprehensive description of the demographic, epidemiologic, and health system context in Kenya, with particular attention to the eight high-burden counties where the Malaria Vaccine Implementation Programme (MVIP) was launched. It highlights the organization of the health system, the burden of malaria, ongoing control strategies, and the strength of Kenya’s immunization infrastructure as the backbone for vaccine introduction. By documenting the epidemiologic, health system, and immunization context, this paper provides the foundation for interpreting RTS,S/AS01 malaria vaccine implementation and evaluation findings and offers lessons for vaccine introduction in other malaria-endemic settings.
Uganda is one of the high burden malaria endemic countries in sub-Saharan Africa (SSA), with women and children under-five disproportionately affected. The objective of this study was to investigate mothers’ willingness to vaccinate their children against malaria in Uganda. A cross-sectional study was conducted for 5281 mothers aged 15–49 years using the 2024–25 Uganda Malaria Indicator Survey (MIS) data. Mothers’ willingness to vaccinate their children against malaria were examined using percentages, the chi-square test, and a multivariable binary logistic regression model. All estimates account for the complex survey design through sampling weights, clustering and stratification. The multivariable model was fitted on 4951 women with complete data on all covariates. Statistical significance was determined at p < 0.05. A weighted prevalence of 92.7
Malaria continues to be endemic within Haiti, despite numerous strategies to reduce its incidence. In 2012, malaria surveillance in clinical settings shifted away from microscopy to HRP2 based Rapid Diagnostic Tests (RDTs) as the primary form of screening. While this approach removes obstacles and delays associated with seeking skilled microscopy, the global emergence of malaria parasites containing HRP deletions has raised concerns of selective pressure and reduced sensitivity. Stored RDTs from febrile patients presenting to clinics in the Ouest and Sud Departments of Haiti were collected between 2021–2023. Total nucleic acid (TNA) was extracted from the entire RDT test strip. TNA was tested for Plasmodium spp. 18S rRNA. Samples testing positive by the 18S rRNA were tested with a triplex assay targeting P. falciparum-specific genes Pfrnr2e2, Pfhrp2, and Pfhrp3. Only samples with successful amplification of Pfrnr2e2 were assessed for Pfhrp2 and/or Pfhrp3 deletions. Of the 2,073 RDT samples analyzed, 193 were positive for Plasmodium spp. 18S rRNA, 49 of which came from negative RDTs (2.61
Malaria diagnosis in Uganda rests almost entirely on rapid diagnostic tests (RDTs) detecting histidine rich protein 2 (HRP2), an antigen expressed only by Plasmodium falciparum. Infections with non-falciparum species are therefore invisible to the national testing algorithm by design. Using a subsample of 466 children from the 2013–14 Democratic Republic of the Congo Demographic and Health Survey, Doctor and colleagues concluded that the risk of missing infections through falciparum-specific RDT use was low, but that conclusion rested on only eight non-falciparum mono-infections. No nationally representative estimate exists for Uganda. We analysed the 2024–25 Uganda Malaria Indicator Survey, in which children aged 6 to 59 months were tested by HRP2 based RDT and by real time PCR with species determination. We estimated survey-weighted prevalence of each species, alone and in mixed infection, and calculated RDT sensitivity separately for P. falciparum mono-infections, mixed infections containing P. falciparum, and non-falciparum mono-infections. Among 6975 children, weighted parasite prevalence by PCR was 17.5
Rodent malaria parasites have long served as essential experimental models for understanding human malaria, yet little is known about their ecology and evolution in the wild. The thicket rat Grammomys poensis (formerly Thamnomys rutilans) is considered the main natural host of murine Plasmodium species in Central Africa. This study aimed to establish a long-term field site to investigate Plasmodium-Grammomys interactions under natural conditions in southeastern Gabon. Over a period of 3 years, trapping was conducted across twelve forest fragments in the Batéké Plateau region using both Tomahawk traps and direct collection at nest. Despite an intensive effort (18,000 trap-nights and 1380 nests inspected), only 20 adult Grammomys were captured, all by collection directly at nest. None were infected with Plasmodium as determined by two sensitive nested-PCR assays targeting the cytb gene, suggesting extremely low transmission or absence of parasite circulation in the study system. These results contrast with earlier studies reporting high infection prevalence in Grammomys from other Central African regions. The absence of infection may reflect local extinction of Plasmodium lineages, limited vector exposure due to the absence of species feeding on arboreal rodents, or host resistance. Because the rodent malaria lines currently used as experimental models in laboratories worldwide were isolated decades ago and have since been maintained through serial passage, access to recent field isolates would be highly valuable for capturing contemporary natural diversity and for enhancing the ecological, evolutionary, and biomedical relevance of these model systems. This study highlights both the challenges of studying this host-parasite system in the wild and the possibility that rodent Plasmodium transmission in this system may be more spatially and ecologically constrained than previously assumed.
Abstract Background The establishment of the invasive malaria vector Anopheles stephensi in Ethiopia threatens existing malaria control efforts. Unlike native vectors, An. stephensi thrives in both urban and rural areas, lacks clear seasonal peaks of abundance, and often breeds in artificial water containers. SumiLarv ™ 2MR containing the larvicide pyriproxyfen (PPF) offers a promising method for controlling the immature stages of An. stephensi in such habitats. This habitat-randomised controlled trial evaluated the efficacy of SumiLarv ™ 2MR against An. stephensi in Danan town, Somali region, Ethiopia. Methods Entomological surveillance identified 102 large artificial containers (“ birkas ”) which were randomly allocated in a 1:1 ratio to receive either SumiLarv ™ 2MR (treatment arm) or no treatment (control arm). SumiLarv ™ 2MR was introduced into the birkas at a dose of one disc per 200 L of water. Morphological identification was performed on adult mosquitoes emerging from control containers, while molecular analysis was conducted on both morphologically identified adult mosquitoes and dead pupae collected from the control and treatment arm. The residual efficacy of PPF content of the remaining discs was also analysed. Inhibition of adult mosquito emergence was assessed every 2 weeks for 40 weeks to determine the duration of efficacy. Results The number of SumiLarv ™ 2MR remained constant in the treatment arm throughout the study period. No significant differences were observed in water volume, pH, or temperature between the two study arms. Introduction of SumiLarv ™ 2MR resulted in complete (100%) inhibition of adult emergence throughout the 40 weeks’ study period (t = 6.26, p < 0.05). Morphological identification of a sub-samples of adult mosquitoes emerging from control containers at different time points confirmed the presence of An. stephensi (99.85%) and An. gambiae s.l. (0.15%). Molecular analysis of morphologically identified adults from control containers and dead pupae from treatment containers confirmed that 98.91% were An.stephensi . Post-trial analysis showed that approximately 82.20% of the original PPF had been released from SumiLarv ™ 2MR during study period. Conclusions The high and sustained efficacy of SumiLarv ™ 2MR demonstrated in this study suggests that it is a suitable tool for controlling An. stephensi in large artificial water storage containers.
Timely treatment of malaria is critical to preventing severe disease and onward transmission. However, treatment-seeking behavior varies across populations and geographic settings. This study examined treatment-seeking time and its determinants among patients with confirmed malaria in Thailand using national surveillance data. A retrospective analysis was conducted using surveillance data from laboratory-confirmed malaria cases recorded in Thailand’s Electronic Malaria Information System (eMIS) from January 2012 to December 2024. The outcome was treatment-seeking time, defined as the number of days between the self-reported date of symptom onset and the date of confirmed malaria diagnosis recorded in the surveillance system. Descriptive statistics summarized treatment-seeking delays, while geographic variation was visualized using choropleth maps. Factors associated with treatment-seeking time were examined using negative binomial regression, with incidence rate ratios (IRRs) and 95
Anopheles funestus is one of the major malaria vectors in sub-Saharan Africa. In Senegal, it is found across nearly all biogeographical areas, often replacing An. gambiae s.l. and sustaining malaria transmission during the dry season. Despite its recognized role as a major vector, the specific contribution of An. funestus to malaria transmission in many malaria-endemic areas remains underexplored. This study aims to assess the contribution of An. funestus in malaria transmission in two health districts in Central Senegal. Adult mosquitoes were collected from July 2018 to November 2019 in the health districts of Nioro (Ndramé Ndimb) and Ndoffane (Tawa Mboudaye) using Human Landing Catches (HLCs) and Pyrethrum Spray Catches (PSCs). Anopheline specimens were identified morphologically using standard dichotomous keys, with species confirmation performed by polymerase chain reaction (PCR). Plasmodium falciparum infection and blood meals sources were determined using Enzyme-Linked Immunosorbent Assay (ELISA) techniques. A total of 6,932 Anopheles mosquitoes were collected. Of these, 73.7
The effectiveness of long-lasting insecticidal nets (LLINs) depends on both chemical durability and sustained biological efficacy under operational conditions. However, insecticide depletion over time does not always correspond directly to loss of entomological effectiveness. This study assessed the relationship between deltamethrin content and biological efficacy of Yorkool® LLINs over 18 months of household use in Benin. A longitudinal study was conducted in southern Benin to monitor Yorkool® LLINs at baseline (T0), 7 months (T7), 12 months (T12), and 18 months (T18). Household practice data were collected and analyzed at all time points except T0. However, chemical and biological evaluations were performed only at T0, T12, and T18. Deltamethrin surface concentrations were quantified using a portable C-Vue HPLC device and expressed in mg/m2. Biological efficacy was assessed using WHO cone bioassays with susceptible Anopheles gambiae Kisumu mosquitoes, measuring knock-down at 60 min and mortality at 24 h. Chemical and biological trends were analyzed comparatively. Mean deltamethrin concentration declined progressively from approximately 55 mg/m2 at T0 to 50.7 mg/m2 at T12 and 45.3 mg/m2 at T18. Despite this chemical depletion, cone bioassays showed consistently high knock-down and mortality rates, with most nets meeting WHO criteria for optimal or minimal effectiveness. The relationship between insecticide concentration and mosquito mortality was positive but non-linear, suggesting a biological threshold effect. Yorkool® LLINs maintained high biological efficacy during the first 18 months of use despite moderate chemical depletion. These findings suggest that loss of insecticide content does not immediately translate into loss of functional performance. However, the absence of follow-up up to 36 months and the use of a susceptible mosquito strain limit extrapolation to full operational lifespan and resistance settings. Extended durability studies are needed to define the functional failure threshold of LLINs.
