Background After a marked reduction in malaria burden in Cambodia over the last decades, case numbers increased again in 2017–2018. In light of the national goal of malaria elimination by 2025, remaining pockets of high risk need to be well defined and strategies well-tailored to identify and target the persisting burden cost-effectively. This study presents species-specific prevalence estimates and risk stratification for a remote area in Cambodia. Methods A cross-sectional survey was conducted in 17 villages in the high-incidence province Mondulkiri in the dry season (December 2017 to April 2018). 4200 randomly selected participants (2–80 years old) were tested for Plasmodium infection by PCR. Risk of infection was associated with questionnaire-derived covariates and spatially stratified based on household GPS coordinates. Results The prevalence of PCR-detectable Plasmodium infection was 8.3% (349/4200) and was more than twice as high for Plasmodium vivax (6.4%, 268) than for Plasmodium falciparum (3.0%, 125, p < 0.001). 97.8% (262/268) of P. vivax and 92.8% (116/125, p < 0.05) of P. falciparum infections were neither accompanied by symptoms at the time of the interview nor detected by microscopy or RDT. Recent travels to forest sites (aOR 2.17, p < 0.01) and forest work (aOR 2.88, p < 0.001) were particularly strong risk factors and risk profiles for both species were similar. Large village-level differences in prevalence of Plasmodium infection were observed, ranging from 0.6% outside the forest to 40.4% inside. Residing in villages at the forest fringe or inside the forest compared to outside was associated with risk of infection (aOR 2.14 and 12.47, p < 0.001). Villages inside the forest formed spatial hotspots of infection despite adjustment for the other risk factors. Conclusions Persisting pockets of high malaria risk were detected in forested areas and in sub-populations engaging in forest-related activities. High levels of asymptomatic infections suggest the need of better case detection plans and the predominance of P. vivax the implementation of radical cure. In villages inside the forest, within-village exposure was indicated in addition to risk due to forest activities. Village-level stratification of targeted interventions based on forest proximity could render the elimination efforts more cost-effective and successful.

In the Greater Mekong Subregion, malaria cases have significantly decreased but little is known about the vectors or mechanisms responsible for residual malaria transmission. We analysed a total of 3920 Anopheles mosquitoes collected during the rainy and dry seasons from four ecological settings in Cambodia (villages, forested areas near villages, rubber tree plantations and forest sites). Using odor-baited traps, 81% of the total samples across all sites were collected in cow baited traps, although 67% of the samples attracted by human baited traps were collected in forest sites. Overall, 20% of collected Anopheles were active during the day, with increased day biting during the dry season. 3131 samples were identified morphologically as 14 different species, and a subset was also identified by DNA amplicon sequencing allowing determination of 29 Anopheles species. The investigation of well characterized insecticide mutations (ace-1, kdr, and rdl genes) indicated that individuals carried mutations associated with response to all the different classes of insecticides. There also appeared to be a non-random association between mosquito species and insecticide resistance with Anopheles peditaeniatus exhibiting nearly fixed mutations. Molecular screening for Plasmodium sp. presence indicated that 3.6% of collected Anopheles were positive, most for P. vivax followed by P. falciparum. These results highlight some of the key mechanisms driving residual human malaria transmission in Cambodia, and illustrate the importance of diverse collection methods, sites and seasons to avoid bias and better characterize Anopheles mosquito ecology in Southeast Asia.

Malaria control is an evolving public health concern, especially in times of resistance to insecticides and to antimalarial drugs, as well as changing environmental conditions that are influencing its epidemiology. Most literature demonstrates an increased risk of malaria transmission in areas of active deforestation, but knowledge about the link between land cover evolution and malaria risk is still limited in some parts of the world. In this study, we discuss different methods used for analysing the interaction between deforestation and malaria, then highlight the constraints that can arise in areas where data is lacking. For instance, there is a gap in knowledge in Cambodia about components of transmission, notably missing detailed vector ecology or epidemiology data, in addition to incomplete prevalence data over time. Still, we illustrate the situation by investigating the evolution of land cover and the progression of deforestation within a malaria-endemic area of Cambodia. To do so, we investigated the area by processing high-resolution satellite imagery from 2018 (1.5 m in panchromatic mode and 6 m in multispectral mode) and produced a land use/land cover map, to complete and homogenise existing data from 1988 and from 1998 to 2008 (land use/land cover from high-resolution satellite imagery). From these classifications, we calculated different landscapes metrics to quantify evolution of deforestation, forest fragmentation and landscape diversity. Over the 30-year period, we observed that deforestation keeps expanding, as diversity and fragmentation indices globally increase. Based on these results and the available literature, we question the mechanisms that could be influencing the relationship between land cover and malaria incidence and suggest further analyses to help elucidate how deforestation can affect malaria dynamics.

ABSTRACT. Gaining an in-depth understanding of malaria transmission requires integrated, multifaceted research approaches. The Asia-Pacific International Center of Excellence in Malaria Research (ICEMR) is applying specifically developed molecular and immunological assays, in-depth entomological assessments, and advanced statistical and mathematical modeling approaches to a rich series of longitudinal cohort and cross-sectional studies in Papua New Guinea and Cambodia. This is revealing both the essential contribution of forest-based transmission and the particular challenges posed by Plasmodium vivax to malaria elimination in Cambodia. In Papua New Guinea, these studies document the complex host–vector–parasite interactions that are underlying both the stunning reductions in malaria burden from 2006 to 2014 and the significant resurgence in transmission in 2016 to 2018. Here we describe the novel analytical, surveillance, molecular, and immunological tools that are being applied in our ongoing Asia-Pacific ICEMR research program.

Global Positioning System (GPS) technology is an effective tool for quantifying individuals’ mobility patterns and can be used to understand their influence on infectious disease transmission. In Cambodia, mobility measurements have been limited to questionnaires, which are of limited efficacy in rural environments. In this study, we used GPS tracking to measure the daily mobility of Cambodian forest goers, a population at high risk of malaria, and developed a workflow adapted to local constraints to produce an optimal dataset representative of the participants’ mobility. We provide a detailed assessment of the GPS tracking and analysis of the data, and highlight the associated difficulties to facilitate the implementation of similar studies in the future.