Baseline genetic characterization of malaria vector populations provides critical data for evidence-based surveillance in persistent transmission foci. This pilot study generated preliminary genetic baseline data for Anopheles populations in the Menoreh Hills border region between Central Java and Yogyakarta provinces, Indonesia, addressing a key geographic gap in regional vector research. Adult female mosquitoes were collected from 3 houses with reported malaria cases in Ngadirejo Village using standardized entomological methods, including human landing, animal landing, and resting collections. Specimens were morphologically identified and molecularly characterized via ITS2 gene sequencing. Phylogenetic analyses were assessed using maximum likelihood methods, and genetic diversity indices were calculated to examine population structure. A total of 62 specimens representing 3 species were collected exclusively through animal landing collections: Anopheles vagus (48 specimens, 77.4%), Anopheles maculatus (9 specimens, 14.5%), and Anopheles kochi (5 specimens, 8.1%). An. kochi exhibited high haplotype diversity (Hd=0.709) with low nucleotide diversity (π=0.004), while An. maculatus showed lower haplotype diversity (Hd=0.480) and higher nucleotide diversity (π=0.026). Phylogenetic analysis revealed Purworejo specimens clustered with regional populations: An. kochi grouped within Clade I with Indonesian isolates; An. maculatus distributed across multiple clades; An. vagus formed a cohesive unit with other Indonesian populations. The exclusive success of animal landing collections in the Menoreh Hills highlands provides key methodological insights. This study offers essential baseline reference data, validates cost-effective genetic surveillance approaches, and supports future large-scale population connectivity studies across the Menoreh Hills malaria transmission complex.
Ernest Mazigo, Hojong Jun, Wang-Jong Lee, Johnsy Mary Louis, Fadhila Fitriana, Jadidan Hada Syahada, Fauzi Muh, Feng Lu, Md Atique Ahmed, Seok Ho Cha, Wanjoo Chun, Won Sun Park, Se Jin Lee, Sunghun Na, Joon-Hee Han, Nyalali Kija, Smart Geodfrey, Eun-Teak Han, Jim Todd, Alphaxard Manjurano, Winifrida Kidima, Jin-Hee Han
Parasites Hosts Dis 2025;63(1):57-65. Published online February 25, 2025
As many countries implement different programs aimed at eliminating malaria, attention should be given to asymptomatic carriers that may interrupt the progress. This was a community-based cross-sectional study conducted in Tanzania from December 2022 to July 2023 within 4 villages from each of the 3 regions, Geita and Kigoma, which are high malaria transmission, and Arusha, which is low transmission. Malaria was diagnosed in asymptomatic individuals aged 1 year and older using the malaria rapid diagnostic test and light microscope. A total of 2,365 of 3,489 (67.9%) participants were enrolled from high-transmission villages. The overall prevalence was 25.5% and 15.8% by malaria rapid diagnostic test and light microscope, respectively. Using the respective tools, the prevalence was significantly higher at 35.6% (confidence interval (CI)=23.6–49.9) and 23.1% (CI=16.2–35.1) in the high-transmission regions (Geita and Kigoma) compared with 2.9% (CI=1.1–3.5) and 1.1% (CI=0.7–1.8) in the low-transmission region (Arusha). Children younger than 15 years and males accounted for the greatest proportion of infections. In the study area, the prevalence of asymptomatic cases was higher than that of reported symptomatic cases in health facilities. We hypothesize that these parasite reservoirs may contribute to the persistence of malaria in the country. Therefore, to achieve comprehensive malaria control in the country, the surveillance and screening of asymptomatic malaria cases are vital.
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Micro-geographic variation in antigenic diversity of PfEBA-175 region II in asymptomatic Plasmodium falciparum infections in Tanzania Jadidan Hada Syahada, Wang-Jong Lee, Hojong Jun, Johnsy Mary Louis, Fadhila Fitriana, Fauzi Muh, Feng Lu, Md Atique Ahmed, Sunghun Na, Wanjoo Chun, Won Sun Park, Bo-Young Jeon, Eun-Teak Han, Jim Todd, Alphaxard Manjurano, Winifrida Kidima, Ernest Mazigo, Frontiers in Immunology.2025;[Epub] CrossRef
The computer vision diagnostic approach currently generates several malaria diagnostic tools. It enhances the accessible and straightforward diagnostics that necessary for clinics and health centers in malaria-endemic areas. A new computer malaria diagnostics tool called the malaria scanner was used to investigate living malaria parasites with easy sample preparation, fast and user-friendly. The cultured Plasmodium parasites were used to confirm the sensitivity of this technique then compared to fluorescence-activated cell sorting (FACS) analysis and light microscopic examination. The measured percentage of parasitemia by the malaria scanner revealed higher precision than microscopy and was similar to FACS. The coefficients of variation of this technique were 1.2-6.7% for Plasmodium knowlesi and 0.3-4.8% for P. falciparum. It allowed determining parasitemia levels of 0.1% or higher, with coefficient of variation smaller than 10%. In terms of the precision range of parasitemia, both high and low ranges showed similar precision results. Pearson’s correlation test was used to evaluate the correlation data coming from all methods. A strong correlation of measured parasitemia (r2=0.99, P<0.05) was observed between each method. The parasitemia analysis using this new diagnostic tool needs technical improvement, particularly in the differentiation of malaria species.
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The discovery and understanding of antigenic proteins are essential for development of a vaccine against malaria. In Plasmodium falciparum, Pf92 have been characterized as a merozoite surface protein, and this protein is expressed at the late schizont stage, but no study of Pv92, the orthologue of Pf92 in P. vivax, has been reported. Thus, the protein structure of Pv92 was analyzed, and the gene sequence was aligned with that of other Plasmodium spp. using bioinformatics tools. The recombinant Pv92 protein was expressed and purified using bacterial expression system and used for immunization of mice to gain the polyclonal antibody and for evaluation of antigenicity by protein array. Also, the antibody against Pv92 was used for subcellular analysis by immunofluorescence assay. The Pv92 protein has a signal peptide and a sexual stage s48/45 domain, and the cysteine residues at the N-terminal of Pv92 were completely conserved. The N-terminal of Pv92 was successfully expressed as soluble form using a bacterial expression system. The antibody raised against Pv92 recognized the parasites and completely merged with PvMSP1-19, indicating that Pv92 was localized on the merozoite surface. Evaluation of the human humoral immune response to Pv92 indicated moderate antigenicity, with 65% sensitivity and 95% specificity by protein array. Taken together, the merozoite surface localization and antigenicity of Pv92 implicate that it might be involved in attachment and invasion of a merozoite to a new host cell or immune evasion during invasion process.
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In the era of (pre) elimination setting, the prevalence of malaria has been decreasing in most of the previously endemic areas. Therefore, effective cost- and time-saving validated pooling strategy is needed for detection of malaria in low transmission settings. In this study, optimal pooling numbers and lowest detection limit were assessed using known density samples prepared systematically, followed by genomic DNA extraction and nested PCR. Pooling strategy that composed of 10 samples in 1 pool, 20 ?l in 1 sample, was optimal, and the parasite density as low as 2 p/?l for both falciparum and vivax infection was enough for detection of malaria. This pooling method showed effectiveness for handling of a huge number of samples in low transmission settings (<9% positive rate). The results indicated that pooling of the blood samples before DNA extraction followed by usual nested PCR is useful and effective for detection of malaria in screening of hidden cases in low-transmission settings.
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