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Background Despite the progress made in this decade towards malaria elimination, it remains a significant public health concern in India and many other countries in South Asia and Asia Pacific region. Understanding the historical trends of malaria incidence in relation to various commodity and policy interventions and identifying the factors associated with its occurrence can inform future intervention strategies for malaria elimination goals. Methods This study analysed historical malaria cases in India from 1990 to 2022 to assess the annual trends and the impact of key anti-malarial interventions on malaria incidence. Factors associated with malaria incidence were identified using univariate and multivariate linear regression analyses. Generalized linear, smoothing, autoregressive integrated moving averages (ARIMA) and Holt’s models were used to forecast malaria cases from 2023 to 2030. Results The reported annual malaria cases in India during 1990–2000 were 2.38 million, which dropped to 0.73 million cases annually during 2011–2022. The overall reduction from 1990 (2,018,783) to 2022 (176,522) was 91%. The key interventions of the Enhanced Malaria Control Project (EMCP), Intensified Malaria Control Project (IMCP), use of bivalent rapid diagnostic tests (RDT-Pf/Pv), artemisinin-based combination therapy (ACT), and involvement of the Accredited Social Health Activists (ASHAs) as front-line workers were found to result in the decline of malaria significantly. The ARIMA and Holt’s models projected a continued decline in cases with the potential for reaching zero indigenous cases by 2027–2028. Important factors influencing malaria incidence included tribal population density, literacy rate, health infrastructure, and forested and hard-to-reach areas. Conclusions Studies aimed at assessing the impact of major commodity and policy interventions on the incidence of disease and studies of disease forecasting will inform programmes and policymakers of steps needed during the last mile phase to achieve malaria elimination. It is proposed that these time series and disease forecasting studies should be performed periodically using granular (monthly) and meteorological data to validate predictions of prior studies and suggest any changes needed for elimination efforts at national and sub-national levels.

The genetic diversity in Plasmodium falciparum antigens is a major hurdle in developing an effective malaria vaccine. Protective efficacy of the vaccine is dependent on the polymorphic alleles of the vaccine candidate antigens. Therefore, we investigated the genetic diversity of the potential vaccine candidate antigens i.e. msp-1, msp-2, glurp, csp and pfs25 from field isolates of P.falciparum and determined the natural immune response against the synthetic peptide of these antigens. Genotyping was performed using Sanger method and size of alleles, multiplicity of infection, heterogeneity and recombination rate were analyzed. Asexual stage antigens were highly polymorphic with 55 and 50 unique alleles in msp-1 and msp-2 genes, respectively. The MOI for msp-1 and msp-2 were 1.67 and 1.28 respectively. A total 59 genotype was found in glurp gene with 8 types of amino acid repeats in the conserved part of RII repeat region. The number of NANP repeats from 40 to 44 was found among 55% samples in csp gene while pfs25 was found almost conserved with only two amino acid substitution site. The level of genetic diversity in the present study population was very similar to that from Asian countries. A higher IgG response was found in the B-cell epitopes of msp-1 and csp antigens and higher level of antibodies against csp B-cell epitope and glurp antigen were recorded with increasing age groups. Significantly, higher positive responses were observed in the csp antigen among the samples with ≥42 NANP repeats. The present finding showed extensive diversity in the asexual stage antigens.

Rapid diagnostic tests (RDTs) are crucial for diagnosing malaria in resource-limited settings. These tests, which detect the histidine-rich protein 2 (PfHRP2) and its structural homologue PfHRP3, are specifically designed to identify Plasmodium falciparum . Deletion of the Pfhrp2 gene in parasite has been reported in India and other malaria-endemic countries. Therefore, periodic surveillance of Pfhrp2 and Pfhrp3 genetic deletions is crucial. We conducted a study to examine these gene deletions in P . falciparum isolates from nine malaria-endemic states in India. In this study, we analyzed 1,558 samples that were microscopically confirmed to be P . falciparum positive. We isolated genomic DNA from all the aforementioned samples, followed by PCR amplification of the Pfhrp2/3 gene. The results showed that the deletion rates for Pfhrp2 and Pfhrp3 genes were 0.44% and 1.47%, respectively. These findings indicate that the gene deletions in all nine states are at low level. Despite these low deletion rates, continuous surveillance is crucial to monitor the efficiency of HRP2 based malaria RDTs. It is recommend that conducting large-scale studies which include other endemic states in India to gain a more comprehensive understanding of the prevalence and impact of these gene deletions over time. This ongoing surveillance will ensure that diagnostic strategies remain effective and that any emerging trends in gene deletions are promptly addressed to achieve the malaria control and elimination.

Background Malaria remains a significant global health challenge, requiring diagnostic approaches that are rapid, cost-effective, and accurate. The present study proposes a hybrid deep learning framework for malaria diagnosis using thin blood smear cell images obtained from the National Institute of Malaria Research (NIMR), a leading research institution of the Indian Council of Medical Research (ICMR). Methods A total of 15,938 thin blood smear microscopic images were preprocessed to enhance the visibility of cellular structures and suppress background noise. Segmentation was performed using the Otsu thresholding method, which automatically determines the optimal threshold to maximize cell contrast and improve feature extraction. The processed images were then used to train the proposed Hybrid Inception-v3 convolutional neural network (CNN), specifically designed for automated cell classification. To establish the robustness of the proposed CNN, comparative and statistical analyses were performed against conventional diagnostic techniques and other state-of-the-art machine learning models. Results The proposed hybrid CNN model achieved a training accuracy of 98.9%, sensitivity of 97.3%, specificity of 99.9%, along with high F 1 score and area under the curve (AUC) values. These results were obtained using the ICMR-NIMR microscopic image dataset, demonstrating the robustness and generalizability of our approach. The model consistently outperformed other evaluated methods, including state-of-the-art machine learning classifiers, and demonstrated performance comparable to or better than conventional microscopy-based examinations. Conclusions The hybrid deep learning framework demonstrated robust performance for malaria diagnosis using microscopic images, suggesting potential to reduce reliance on expert microscopists. These findings highlight its potential to improve diagnostic precision and strengthen malaria control efforts, especially in resource-limited settings. Graphical Abstract

Background Malaria is known as a disease of poverty because of its dominance in poverty-stricken areas. Madhya Pradesh state in central India is one of the most vulnerable states for malaria morbidity and mortality. Socio-economic, environmental and demographic factors present challenges in malaria control and elimination. As part of the Malaria Elimination Demonstration Project in the tribal district of Mandla in Madhya Pradesh, this study was undertaken to assess the role of different social-economic factors contributing to malaria incidence. Methods The study was conducted in the 1233 villages of district Mandla, where 87% population resides in rural areas. The data was collected using the android based mobile application—SOCH for a period of 2 years (September 2017 to August 2019). A wealth index was computed along with analysis of the socio-economic characteristics of houses with malaria cases. Variables with significant variation in malaria cases were used in logistic regression. Results More than 70% of houses in Mandla are Kuccha (made of thatched roof or mud), 20% do not have any toilet facilities, and only 11% had an annual income of more than 50,000 INR, which converts to about $700 per year. Households with younger heads, male heads, more number of family members were more likely to have malaria cases. Kuccha construction, improper water supply, low household income houses were also more likely to have a malaria case and the odds doubled in houses with no toilet facilities. Conclusion Based on the results of the study, it has been found that there is an association between the odds of having malaria cases and different household variables such as age, gender, number of members, number of rooms, caste, type of house, toilet facilities, water supply, cattle sheds, agricultural land, income, and vector control interventions. Therefore, a better understanding of the association of various risk factors that influence the incidence of malaria is required to design and/or deploy effective policies and strategies for malaria elimination. The results of this study suggest that appropriate economic and environmental interventions even in low-income and poverty-stricken tribal areas could have huge impact on the success of the national malaria elimination goals.

Background Compared to 2017, India achieved a significant reduction in malaria cases in 2020. Madhya Pradesh (MP) is a tribal dominated state of India with history of high malaria burden in some districts. District Mandla of MP state showed a considerable decline in malaria cases between 2000 and 2013, except in 2007. Subsequently, a resurgence of malaria cases was observed during 2014 and 2015. The Malaria Elimination Demonstration Project (MEDP) was launched in 2017 in Mandla with the goal to achieve zero indigenous malaria cases. This project used: (1) active surveillance and case management using T4 (Track fever, Test fever, Treat patient, and Track patient); (2) vector control using indoor residual sprays and long-lasting insecticidal nets; (3) information education communication and behaviour change communication; and (4) regular monitoring and evaluation with an emphasis on operational and management accountability. This study has investigated malaria prevalence trends from 2008 to 2020, and has predicted trends for the next 5 years for Mandla and its bordering districts. Methods The malaria prevalence data of the district Mandla for the period of January 2008 to August 2017 was obtained from District Malaria Office (DMO) Mandla and data for the period of September 2017 to December 2020 was taken from MEDP data repository. Further, the malaria prevalence data for the period of January 2008 to December 2020 was collected from DMOs of the neighbouring districts of Mandla. A univariate time series and forecast analysis was performed using seasonal autoregressive integrated moving average model. Findings Malaria prevalence in Mandla showed a sharp decline [− 87% (95% CI − 90%, − 84%)] from 2017 to 2020. The malaria forecast for Mandla predicts zero cases in the next 5 years (2021–2025), provided current interventions are sustained. By contrast, the model has forecasted a risk of resurgence of malaria in other districts in MP (Balaghat, Dindori, Jabalpur, Seoni, and Kawardha) that were not the part of MEDP. Conclusion The interventions deployed as part of MEDP have resulted in a sustainable zero indigenous malaria cases in Mandla. Use of similar strategies in neighbouring and other malaria-endemic districts in India could achieve similar results. However, without adding extra cost to the existing intervention, sincere efforts are needed to sustain these interventions and their impact using accountability framework, data transparency, and programme ownership from state to district level.

Background Many malaria endemic countries are heading towards malaria elimination through the use of case management and vector control strategies, which employ surveillance, improving access to early diagnosis, prompt treatment., and integrated vector control measures. There is a consensus that elimination of malaria is feasible when rapid detection and prompt treatment is combined with mosquito-human contact interruption in an efficient and sustainable manner at community levels. This paper describes results of an integrated case management and vector control strategy for reducing malaria cases in 1233 villages over 3 years in district Mandla, Madhya Pradesh, India. Methods The project enrolled the entire population (1,143,126) of Mandla district for fever surveillance followed by testing of febrile cases and treatment of positive subjects using T4 strategy, which is Track (by fever), Test (by RDTs), Treat (by ACT) and Track (for completion of treatment). In addition to the active and passive surveillance for detection and treatment of febrile cases, the project conducted mass screening and treatment to clear the asymptomatic reservoirs of infection. Febrile cases were also tested in the out-patient department of the District Hospital from June 2018 to September, 2018 and in a community-based medical camp from November 7 to 14, 2019. The project also used vector control measures for interrupting human-mosquito contact, and information, education and communication (IEC) campaigns to increase demand for malaria services at community level. Results This project has revealed about 91% reduction of indigenous cases of malaria during the period from June 2017 to May 2020, through case management and vector control strategies. A total 357,143 febrile cases were screened, out of which 0.19% were found positive for the presence of malaria parasites, with Plasmodium falciparum and Plasmodium vivax ratio of 62:38. The prevalence of malaria was higher in individuals > 15 years of age (69% cases). The positivity rate was 0.33% in 2017–18, 0.13% in 2018–19, and 0.06% in 2019–20. In all of the 3 years of the project, the peak transmission correlated with rains. Mass screening revealed 0.18% positivity in Sep-Oct 2018, followed by 0.06% in June 2019, and 0.03% in December 2019, and these were mostly asymptomatic cases in the community. Imported cases into the district were mostly contributed by the distant state of Telangana (51.13%). Fever patients tested for malaria parasites in the District Hospital and medical camp revealed zero cases. Conclusion Using the current intervention and prevention tools along with optimum utilization of human resources, a 91% reduction in indigenous cases of malaria was seen in the district in 3 years. The reduction was similar in the three high prevalence blocks of the district. These results reveal that malaria elimination is achievable in India within a stipulated time frame. The reduction of malaria at the community level was further validated when zero malaria cases were diagnosed during hospital and community-based studies in Mandla. Prompt detection and treatment of imported/migratory cases may have prevented outbreaks in the district. This project has demonstrated that field programmes backed by adequate technical, management, operational, and financial controls with robust monitoring are needed for achieving malaria elimination.

Background Changes in social, belief, and behavioural practices are essential for the success of any public health delivery programme. In the planning stages of the Malaria Elimination Demonstration Project (MEDP), priority was given to communication with a goal to develop capacity of health workers and to improve the knowledge, attitude and practices (KAP) of the people of Mandla. This paper describes the level of community knowledge on malaria, including its prevention, diagnosis, treatment-seeking behaviour, and the level of satisfaction with the services provided by the project. Methods A cross sectional survey was undertaken in 1233 villages of Mandla to study the KAP and self-assessed improvement in knowledge and satisfaction level of the community. The goal of the study was to understand whether there is need for strengthening communication strategy of MEDP for better impact. The survey was conducted amongst the head/eligible members of the 733 households located in the nine blocks of the district using clustered random sampling. Results Though four-fifths of the respondents were able to correlate the transmission of malaria with mosquitoes, misconceptions existed among them. The types of malaria were not known to everyone. Only 39% were aware of the Indoor Residual Spray (IRS) and 41% understood the value of Long-Lasting Insecticidal Nets (LLIN). Around 71% of subjects surveyed were aware of the proper diagnostic tests for malaria. A total of 87% of the respondents knew about the MEDP staff working in their respective villages. Conclusion The study reported gaps in knowledge on malaria at community level. The self-assessment of the community revealed that the communication strategy established by MEDP in Mandla district has been useful to them as they are becoming better informed about the prevention and treatment aspects of disease. The lessons learned as revealed in the KAP survey will improve malaria elimination outcomes in a timely manner.

Introduction World Health Organization (WHO) in 2010, recommended 0.75 mg/kg single dose primaquine for killing gametocytes after ruling out glucose-6-phosphate dehydrogenase (G6PD) deficiency. However, considering the practical feasibility of G6PD testing, WHO revised the recommendation in 2012 to 0.25 mg/kg single dose primaquine on day-1. This study aims to investigate the efficacy and safety of single-low dose primaquine (0.25 mg/kg) administered on day-1 to intervention arm compared to standard regimen, high-dose primaquine (0.75 mg/kg) administered on day-2 to control arm. Methods and analysis This is a hospital based, open-label, multi-centric, randomized-controlled trial being conducted at four different sites in India, with a target sample size of 496 i.e., 124 participants per site. Participants above 18 years of age with uncomplicated P. falciparum malaria and normal glucose-6-phosphate dehydrogenase (G6PD) activity, defined as > 4.0 IU/g Hb using the point-of-care test, weight > 40 kg, haemoglobin > 8 g/dL are treated with ACT and randomized to either control or intervention arm. Efficacy assessment is based on gametocytemia as inferred from examination of stained peripheral blood smears during enrolment and follow-up. In addition, quantitative nucleic acid sequence-based amplification (QT-NASBA) for Pf s25 mRNA, will be used to quantify the gametocytes on day 1, 2/3, 7, and 14. Safety assessment during 14-day follow-up will be based on percentage of participants with more than 25% drop in haemoglobin, haemoglobin drop of 5 g/dl, haemoglobin value less than 8 g/dl, mean of maximum drop in haemoglobin during 28-day follow-up; and frequency of adverse events reported during the follow-up in both the arms. The primary outcome of the study is based on gametocyte density and point prevalence of gametocyte (efficacy) and haemoglobin concentration (safety) from enrolment to day 14 (for efficacy) and day 28 (for safety). Ethics and dissemination Study has been approved by Institutional Ethics Committee (NIMR/IEC-M/2023/970/v6-Jun/01). The study findings will be disseminated through publications in peer-reviewed journals as well as academic presentations. The findings will support evidence-based recommendations for national malaria control programs in India, potentially advocating for the adoption of the lower dose primaquine regimen. Trial registration number CTRI/2025/07/091988 (CTRI registration). Trial status Enrollment started in August, 2025. The trial is going on.

Background Mass screening and treatment (MSaT) aims at reducing the spread of malaria in communities by identifying and treating infected persons regardless of the symptoms. This study was conducted to identify and treat asymptomatic cases using MSaT approaches in the community. Methods Three rounds of MSaT using cluster combination approaches were carried out during September 2018 to December 2019 to identify and treat asymptomatic malaria cases in the community. All individuals who were present in the household were screened using RDT irrespective of malaria related symptoms. Simultaneously thick and thin blood smear and blood spot were collected for further analysis using microscopy and diagnostic PCR done in a subset of the samples. Results Logistic regression analysis revealed that asymptomatic malaria cases significantly less among the older age groups compared with < 5 years children (OR ranged between 0.52 and 0.61; p < 0.05), lowest in cluster 4 (OR = 0.01; p < 0.0001); during third round of MSaT survey (OR = 0.11; p < 0.0001) and significantly higher in moderate to high endemic areas (OR = 88.30; p < 0.0001). Conclusion Over the three rounds of MSaT, the number of asymptomatic cases were significantly less in the older age groups, and during third round. Similarly, the asymptomatic cases were significantly less in the low endemic area with API < 1 (cluster four). Therefore, the malaria elimination programme may consider the MSaT strategy to identify asymptomatic cases that would be otherwise missed by routine fever based surveillance. This MSaT strategy would help accomplish the malaria elimination goal in an expedited manner.