Plasmodium; of all the parasitic diseases, malaria has the highest prevalence and mortality with an estimated 247 million cases and 619,000 deaths recorded worldwide as of 2021. Malaria causes febrile illness with several changes in blood cell parameters. Some of these changes include leucopenia, thrombocytopenia, and anaemia. If these changes could be correlated with the degree of parasitaemia, it can serve as a guide to physicians when treating malaria. This study was therefore aimed at correlating haematological parameters with levels of parasitaemia during malaria infection.Malaria is a parasitic disease caused by various species of the blood parasite The study was a cross-sectional study involving 89 malaria positive patients. About 5 ml of blood was collected from each participant who gave his or her informed consent to partake in the study. A full blood count was performed on their samples to determine their haematological parameters using a haematology auto-analyzer. A parasite count was also performed via microscopy to determine the degree of parasitaemia. The data obtained from the study was entered into a database and statistically analysed using Statistical Package for Social Sciences (SPSS) version 23 and Microsoft Excel 2016.The study comprised of 89 participants out of which 35 were males and 54 were females with the mean age of 26.15 years. Secondary education participants were the highest with quaternary education the lowest. The highest parasite count recorded was 398,174 parasites/µl of blood, lowest count was 101 with the average being 32,942.32584. There was also a significant positive Pearson’s correlation between total WBC and parasitaemia and with the WBC differentials, neutrophils, lymphocytes and monocytes had positive correlations while eosinophils and basophils had negative correlations. Furthermore, platelets, total RBC’s, haemoglobin, MCH, MCHC and Hct all showed negative correlations. Linear regression also showed a linear relationship between parasite density and the various haematological parameters.The linear relationship (correlation) between WBC and MCH were the only significant ones at 95% and 99% confidence interval, respectively based on a two-tail t-test. Also, based on the regression analysis, the changes caused by WBC and PLT were the only significant changes at 95% confidence level in a two-tailed t-test. Anopheles mosquitoes [Plasmodium falciparum and Plasmodium vivax—pose the greatest threat [Malaria is a life-threatening disease that is caused by parasites transmitted to people through the bites of infected female squitoes . The infsquitoes , 2. Thert threat . Malariat threat . In 2021t threat . In Ghant threat .P. falciparum-infected erythrocytes, particularly those with mature trophozoites, adhere to the vascular endothelium of venular blood vessel walls and do not freely circulate in the blood [All of the clinical symptoms we see in malaria infection are caused by the asexual erythrocytic or blood stage parasites . As the he blood . When thhe blood .Malaria causes acute febrile illness and is often difficult to differentiate from other febrile illnesses such as leptospirosis and arboviral infection hence it might not be detected early in patients resulting in several complications . SymptomP. falciparum infections since the parasite invades red blood cells of all ages hence causing the most severe infections [P. falciparum infections, it would be vital if a correlation between haematological parameters and parasitaemia could be established as it will enable physicians to determine how to proceed with early treatment and management of malaria.Furthermore, when one is infected with malaria, various haematological alterations such as anaemia, thrombocytopenia, leukocytosis or leukopenia do occur , 17. Lymfections . Due to The aim of this study was to correlate changes in blood cell parameters with levels of parasitaemia in malaria infection. This study was a cross sectional study conducted from January 2018 to December 2018 at the Mamprobi polyclinic in Accra, Ghana.The study comprised of eighty-nine (89) male and female patients attending the polyclinic and diagnosed with malaria infection and who gave their written informed consent.Three to five millilitres (3–5 ml) of blood samples were collected from a prominent vein of each participant by venipuncture into lavender-topped tubes containing Ethylene-Diamine-Tetra-Acetic acid (EDTA) and thoroughly mixed to prevent them from clotting. The patients' unique identification numbers on consent forms were written on each tube to ensure confidentiality. A haematology analyzer was then used to perform a full blood count on the patients' blood samples for their haematological parameters. Thick and thin blood films were also prepared using the same patients' blood samples. The thick films were stained with Giemsa and the thin films with Leishman stain. The thick films were then examined under × 100 magnification for malaria parasites identification and the thin films for parasite specie identification. To determine the parasite density, two hundred (200) leukocytes were first counted whilst observing the malaria parasites present. If less than 10 parasites were observed after counting 200 leukocytes the count was continued till 500 leukocytes were counted. The parasite density was then calculated using the formula below.This gave the number of parasites/µl of blood.Data obtained were analyzed using Microsoft Excel 2016 and Statistical Package for Social Sciences (SPSS) software Version 23 . Descriptive statistics, Correlation analysis and linear regression analysis were carried out on the data.There were eighty-nine (89) participants in the study out of which 35 were males and 54 were females representing 39% and 61%, respectively. The oldest participant was 78 years while the youngest was 9 months. The mean age recorded was 26.15 years. With regards to educational background, 13 participants had none, 15 had primary, 43 had secondary, 16 had tertiary and 2 had quaternary education  rise in parasite density there was 0.300 units (WBC × 109/l) rise in WBC and of the five white cells, the neutrophils were the most responsible for this rise by 0.297 units (Neu × 109/l) followed by monocytes and lymphocytes with a rise of 0.053 and 0.014 units respectively. Eosinophils and basophils fell by 0.021 and 0.025 units, respectively anytime parasitaemia rose by 1 unit  and parasitaemia. The model had parasitaemia as the outcome variable and the various haematological parameters as predictor variables. A multiple R of 0.44 (correlation between the outcome and predictor variables) was obtained. This suggests a moderate degree of association. The R-squared of 0.195 implies that approximately 19.5% of variations in parasitaemia (outcome variable) can be explained by the selected Haematological parameters (predictor variables). However, the model’s adjusted R-squared value which explains variation in outcome variables caused by predictor variables while adjusting for the number of predictor variables was relatively low 0.125). This indicates that other unmeasured factors might also be influencing the outcome variable . This indicated that the predictor variables had a significant influence on predicting the outcome variable. The significant F-statistic and corresponding p-value supported the model's overall fit. Notwithstanding, only WBC and platelet counts showed statistically significant relationships with the outcome variable. The positive coefficient for WBC indicates that an elevation in WBC is associated with an increase in the outcome variable. In contrast, the negative Platelet coefficient indicates that a decrease in Platelet count is associated with a decline in the outcome variable. All the other predictor variables  did not show statistically significant associations with parasitaemia.In this study eighty-nine malaria patients were recruited, and the majority of participants (61%) were females, and the rest (39%) were males. The average parasite count of the participants was 32,942.32584 parasites/µl with the lowest being 101 parasites/µl and the highest of 398,174 parasites/µl. The average parasite count of the males  was 50% higher than that of their female counterparts  although, the female participants outnumbered the males in the study. The age range of the participants was between 9 months and 78 years with the mean age being 26.15 years. The age with the highest parasitaemia was 12 while that with the lowest was 62. Also, most of the WBC counts were either within the normal range or lower than the reference but there were few cases of leukocytosis. The haemoglobin, platelet and other RBC indices also showed variations as parasitaemia increased. This clearly demonstrates that there is a relationship between the degree of parasitaemia and various haematological parameters.After the analysis, correlations were established between parasitaemia and various haematological parameters. There was a positive correlation between parasitaemia and WBC count.Plasmodium parasite resides in RBCs, their activities results in the lyses of the cells thus reducing their numbers leading to haemolytic anaemia in most malaria infection cases [Eosinophils and basophils are the least numerous WBCs in circulation and often associated with allergic reactions thus this might be a reason for their negative correlation. Also, haemoglobin and RBC both had negative correlations as parasitaemia increased. This could be due to the fact that as the on cases .MCH and MCHC, which are dependent on RBC, Hb and Hct values also showed negative correlations. There was also a reduction in platelet levels per every unit increase in parasite density, which could be due to the immune and non-immune destruction of platelets during malaria infection. The exact course of platelet destruction is not known but some of the assumed mechanisms may include coagulation disturbances, sequestration in spleen, antibody mediated platelet destruction, oxidative stress, and the role of platelets as cofactors in triggering severe malaria .Plasmodium infested RBCs and normal RBCs can result in fluctuations in Hct [. found that platelets, lymphocytes, eosinophils, red blood cell count and haemoglobin were significantly lower in malaria-infected children [. also carried out a study where they showed that 70% of the patients with malaria had thrombocytopenia, 94% anaemia, 12% lymphopenia and 17% monocytosis [. who found that neutrophils levels were significantly higher in cases of falciparum malaria in comparison to healthy normal subjects [Again, the regression relationship between RBC, Hb and MCH with parasite density were negative while that of MCHC and Hct were positive. However, these relationships were insignificant based on a two-tail t-test performed at 95% confidence level. The findings of this study are similar to that of Wang and Xing who posited that interactions between s in Hct . Similarchildren . Again, ocytosis . The cursubjects .P. falciparum malaria parasite are known to adhere to endothelial cells lining blood vessels [It is worthy to mention that red blood cells infected with  vessels . This ph vessels . This ad vessels . Indeed  vessels . In termThe outcome of the study indicates that there exists a correlation between the various haematological parameters and degree of parasitaemia. However, upon performing two-tail t-test to determine the significance of the correlation at 95% and 99% confidence level, it was concluded that only WBC and MCH had a significant linear relationship (correlation) with the degree of parasitaemia at 95% and 99% confidence level, respectively. Regression analysis also showed that the variation in parasitaemia (dependent variable) resulted in changes in the various haematological parameters (independent variables). Howbeit, only the changes caused by WBC and PLT were significant at 95% confidence level in a two-tailed t-test.


Plasmodium infections; however, the findings remain inconclusive. This study synthesized differences in MDA concentrations among patients with different levels of clinical severity, uninfected controls, and different Plasmodium species. The research protocol was registered in PROSPERO (CRD42023393540). Systematic literature searches for relevant studies were performed using the Embase, MEDLINE, Ovid, ProQuest, PubMed, Scopus, and Google Scholar databases. Qualitative and quantitative syntheses  of distinct MDA concentrations between the disease groups were performed. Twenty-three studies met the eligibility criteria and were included in the systematic review. Overall, MDA concentrations were significantly elevated in participants with malaria relative to uninfected controls : 1.88–3.14, I2: 96.22%, 14 studies). Increased MDA concentrations in participants with malaria compared with uninfected controls were found in studies that enrolled patients with P. falciparum malaria  and P. vivax malaria . Our findings confirm that MDA concentrations increase during Plasmodium infection, indicating a rise in oxidative stress and lipid peroxidation. Thus, MDA levels can be a valuable biomarker for evaluating these processes in individuals with malaria. However, further research is necessary to fully elucidate the intricate relationship between malaria, antioxidants, oxidative stress, and the specific role of MDA in the progression of malaria.Several studies have evaluated the relationship between malondialdehyde (MDA) concentrations and  Lipid peroxidation is involved in the pathogenesis of various tissue injuries ,2. OxidaPlasmodium parasites  through bites from Anopheles spp. mosquitoes [Plasmodium infection, host cells release reactive oxygen species (ROS), which not only clear the parasite but also damage host cells and tissues, leading to severe pathologies [In humans, malaria is caused by the infection of one or more of the five squitoes . Malariasquitoes . In resphologies . Many sthologies ,15,16,17hologies ,19 and rhologies ,21. The hologies ,23. Humahologies ,25, whicPlasmodium infections has been investigated in the literature, but the results are inconsistent. Furthermore, MDA concentrations may vary according to the clinical severity of the disease or Plasmodium species. Therefore, this systematic review synthesized differences in MDA concentrations between participants with malaria and uninfected controls. Additionally, we examined variations in MDA levels between patients with severe and nonsevere malaria, as well as between patients with P. falciparum and P. vivax malaria.The relationship between MDA levels and This research protocol was registered in PROSPERO (CRD42023393540). The study was conducted and reported according to the PRISMA protocol for reporting systematic reviews and meta-analyses . SystemaThe eligibility criteria for the review were determined using the patient, intervention, comparison, outcome (PICO) framework  as folloPlasmodium identification methods, and MDA concentration methods. Study selection and data extraction were performed independently by two authors (O.M. and M.K.), and any disagreements were discussed with another author (A.M.).The articles were input into EndNote version 20.0 , duplicates were removed, and the remaining articles were checked for eligibility. After removing irrelevant articles, the full texts of the remaining articles were examined to see if they met the requirements. Studies that failed to meet the eligibility requirements were then removed, and a clear explanation was given. Data on the study and participant characteristics and the PICO criteria were extracted from each study, including the name of the first author, publication year, study design and area, the number of participants and age range, MDA concentrations in malaria patients and uninfected controls, The Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) checklists for observational studies were used to assess the quality of the studies . These cP. falciparum vs. P. vivax infection). Quantitative synthesis  of distinct MDA concentrations between the disease groups  was performed using the random-effect model [2 statistics were utilized to assess between-study heterogeneity, with I2 values <50, 50–75, and >75 categorized as having low, moderate, and high between-study heterogeneity, respectively [Qualitative synthesis was used to explain qualitative differences in MDA concentrations in the disease groups , MEDLINE n = 34), Ovid (n = 282), ProQuest n = 42), PubMed (n = 125), and Scopus n = 167). After removing 283 duplicate articles, the remaining 558 were reviewed, and a further 486 irrelevant articles were removed. The remaining 72 full-text articles were examined against the eligibility criteria, and those that failed to meet them were excluded with a clear explanation, leaving 19 articles [. After r, Ovid , and most study participants were adults (39.1%). Over half of the studies used microscopy alone for Plasmodium detection (52.2%)  were published between 2010 and 2022 and had a cross-sectional design (91.2%). The majority were performed in African countries (41.8%) and Asian countries (41.8%), with Nigeria and India the most representative countries in Africa (63.6%) and Asia (72.7%), respectively. Most studies enrolled patients infected with  (52.2%) .Based on the STROBE checklist, seven studies (21.7%) were of high quality, and the remainder (78.3%) were of moderate quality. None of the studies were of low quality . To inveA qualitative synthesis showed increased MDA concentrations in participants with malaria compared with uninfected controls in 19 studies ,53,54,55p < 0.01, Cohen d: 2.51, 95% confidence interval (CI): 1.88–3.14, I2: 96.22%, 14 studies; Fourteen studies reported quantitative data; thus, they were used for quantitative synthesis ,50,53,54Plasmodium species, assays for Plasmodium identification, and study quality showed that the Plasmodium species and the study’s quality may affect the pooled effect estimate  and those with P. vivax malaria . The subgroup analysis of the study’s quality revealed a borderline subgroup difference  and moderate quality .Meta-regression analyses of publication year, participants, study design, continent, A qualitative synthesis showed increased MDA concentrations in severe and nonsevere malaria patients in a study by Sakyi et al. . RegardiP. falciparum malaria compared with P. vivax malaria. Lower MDA concentrations in P. falciparum malaria compared with P. vivax malaria have been reported [P. falciparum and P. vivax malaria using the meta-analysis approach could not be determined due to the insufficient number of studies.In two studies ,50, a qureported . Differep < 0.01). The trim-and-fill analysis showed increased MDA concentrations in malaria cases compared with uninfected controls .In all reruns using the leave-one-out meta-analysis method, participants with malaria had higher MDA concentrations than the uninfected controls . The funPlasmodium infection in humans results in the host’s release of ROS, which aid in parasite clearance [MDA is a well-known marker of oxidative stress in several diseases ,8,9. It learance . Howeverlearance . The prilearance . As a relearance . Thus, MP. falciparum or P. vivax. Due to the limited number of studies investigating MDA levels in both of these Plasmodium species, the results exhibit significant heterogeneity and lack clarity. Specifically, only two studies [P. falciparum than in P. vivax infections. This disparity may be attributed to the more severe nature of P. falciparum infection, which induces greater oxidative stress in the host cells. However, another study found lower MDA concentrations in P. falciparum malaria than in P. vivax malaria [P. vivax malaria to oxidative stress, potentially due to lower levels of ascorbic acid [Plasmodium species, regardless of parasitemia [Plasmodium infections induce oxidative stress-related MDA buildup.The subgroup meta-analysis revealed elevated MDA concentrations in cases of malaria caused by  studies ,50 repor malaria . This dibic acid . Overallasitemia . The subThe presence of MDA in malaria-infected individuals indicates the occurrence of lipid peroxidation and oxidative stress. Notably, MDA is one of many markers used to assess oxidative stress and lipid peroxidation, and its measurement alone may not provide a comprehensive understanding of the overall oxidative status in malaria. Furthermore, the association between MDA and the antioxidant system is an important aspect to consider in malaria. The antioxidant system is crucial to maintain the balance between ROS production and elimination . It consOur systematic review and meta-analysis had some limitations. First, there was heterogeneity in MDA levels between the studies included in the meta-analysis. This heterogeneity might potentially be influenced by the very wide range of MDA levels due to differences in the quality of the included studies, as demonstrated by the subgroup analysis. Furthermore, MDA levels could be potentially influenced by characteristics of the diseased cohorts, such as the age of participants, the timing, and disease severity. Second, publication bias was observed in the meta-analysis of MDA concentrations between malaria patients and uninfected controls due to the small number of studies.Plasmodium infection and are independent of the Plasmodium species , at least for the limited studies included in this meta-analysis. The measurement of MDA levels can serve as a useful biomarker to evaluate oxidative stress and lipid peroxidation in individuals with malaria. These findings suggest that MDA concentrations can be suitably tracked as a potential indicator of Plasmodium infection. However, further research is necessary to fully understand the relationship between malaria, oxidative stress, and the role of MDA in the disease.Our study confirms that MDA concentrations increase in cases of


Plasmodium spp. Several blood phenotypes have been associated with malarial resistance, which suggests a genetic component to immune protection.Malaria is a deadly disease caused by One hundred and eighty-seven single nucleotide polymorphisms (SNPs) in 37 candidate genes were genotyped and investigated for associations with clinical malaria in a longitudinal cohort of 349 infants from Manhiça, Mozambique, in a randomized controlled clinical trial (RCT) . Malaria candidate genes were selected according to involvement in known malarial haemoglobinopathies, immune, and pathogenesis pathways.TLR4 and related genes with the incidence of clinical malaria (p = 0.0005). These additional genes include ABO, CAT, CD14, CD36, CR1, G6PD, GCLM, HP, IFNG, IFNGR1, IL13, IL1A, IL1B, IL4R, IL4, IL6, IL13, MBL, MNSOD, and TLR2. Of specific interest, the previously identified TLR4 SNP rs4986790 and the novel finding of TRL4 SNP rs5030719 were associated with primary cases of clinical malaria.Statistically significant evidence was found for the association of TLR4, as well as associated genes, in clinical malaria may provide insight into treatment and drug development.These findings highlight a potential central role of TLR4 in clinical malarial pathogenesis. This supports the current literature and suggests that further research into the role of The online version contains supplementary material available at 10.1186/s12936-023-04549-8. Plasmodium genus, particularly Plasmodium falciparum and Plasmodium vivax, which are the two species with a wider health impact. The disease affected approximately 241 million humans worldwide in 2020—a number comparable to that of the COVID-19 global pandemic ), IL1B (rs1143634), IL4R (rs1805010 [I75X] and rs1805015 [S503P]), IL4 (rs4889 [P81L and P81R]), IL13 (rs1800925), MBL (rs11003123 and rs11003124), TLR2 (rs3804099), and TLR4 (rs5030719 [Q510H]).Results from the Poisson analysis correlating primary incidence of malaria to SNPs of interest that were found to be statistically significant  for child and maternal SNPs are listed in Tables CAT (rs769214), G6PD (rs1050829 [N156Y and N156D]), HP (rs5470), ICAM1 (rs5498 [K469E]), IFNGR1 (rs2234711 and rs7749390), IFNG (rs2430561), IL6 (rs2069830 [P32S]), MNSOD (rs4880 [V16A]), TLR2 (rs7656411), and TLR4 (rs5030719 [Q510H]), all of which were still statistically significant after including in the analysis the respective children’s alleles as co-factors. An overlapping set of genes, HP, ICAM1, IFNG, TLR2, and TLR4 were found to contain SNPs statistically significantly associated with malaria in both maternal and child analyses. Additionally, the SNPs rs5498 in ICAM1, rs2430561 in IFNG, and rs5030719 in TLR4 were found in both maternal and child analyses, whereas the other SNPs in HP, and TLR2 differed between mother and child. In both cases of IFNG rs2430561 and TLR4 rs5030719, the risk genotypes in maternal and child analyses were found to be the same. The T variant of SNP rs5030719 as a malarial risk factor is a novel finding previously unreported in the literature. As a measure of quality control, the allelic distribution of all genes was assessed in relation to Hardy–Weinberg equilibrium , IL13 (GCACT and GTGCT), and TLR4 (AT); as well as four haplotypes in three maternal genes, IFNGR1 (AT(TT) and GC(TT)), IL6 (GCT), and TLR4 (GT). The SNP rs values and further information on the haplotypes are provided in Table TLR4 was the only gene identified to be statistically associated with malaria in both child and maternal analysis as well as producing haplotypes associated with the disease in the HTR analysis.Haplotype trend regression of the SNPs identified in Tables TLR4. Further HTR analysis implicates the additional genes ABO, IFNGR1, IL13, and IL6.Primary analysis of these targeted genetic polymorphisms highlights a shortlist of genes that are potentially associated with malaria and have biological plausibility in pathology of the disease. Of these, the only gene identified in both maternal and child analyses was IL) and toll-like receptor (TLR) genes is expected, as in the literature it has been demonstrated that these immunologically related molecules are associated with clinical malaria, ranging across the spectrum of impact from cellular biology to clinical drug and vaccine development [ICAM1 expression, of which we have found evidence for statistical association of IL1, IL4 , IL6, IL13, TLR2, TLR4, and IFNG  polymorphisms with malaria. In this context, it can be argued that these immunogenically related genes, along with the identified gene HP, play a supporting role in inducing ICAM1. Circulating IL6 and IFN-γ levels are associated with clearance of the parasite, though in high concentrations, may manifest as extreme symptoms of malaria [The identification of various interleukin , interferons (IFNG and IFNGR1), and downstream immunomodulatory proteins (CD14 and CD36) fit the biological narrative of this hypothesis, warranting future interrogation into the role and potential use of these cytokines in treatment of malaria.Intermediary immunomodulators, such as TLRs, including TLR4, bridge signalling between Pathogen-Associated Molecular Patterns (PAMPs) and intercellular adaptors such as MyD88 and TIR domain-containing adapter-inducing IFN-β (TRIF), which then have a plethora of intercellular effects, including cytokine signalling . An agnoPlasmodium spp. mature trophozoites and schizonts in cytoadhesion between parasitized red blood cells and endothelial cells lining the local microvasculature, leading to parasite sequestration. This interaction is mediated by the parasitic ligand, P. falciparum erythrocyte membrane protein 1 (PfEMP-1) and possibly other parasite-related proteins [ICAM1 [GCLM, and its associated gene, GCLC [ICAM1 are low in peripheral blood, but they are expressed at higher levels in the liver [GCLC and GCLM in malaria pathology, the glutathione synthesis pathway has a known role in combating clinical malaria , playing a key role in the scavenging of free radicals and alleviating oxidative stress injury. This in turn allows for better maintenance of cellular homeostasis during infection periods, reducing the incidence of cellular damage and consequently, apoptosis [ICAM-1 is a cell-surface receptor utilized by proteins . Althougproteins –35. Thers [ICAM1 , 36–40, ne, GCLC  being pohe liver , suggestpoptosis . Much ofICAM1 SNPs, rs5490, rs5491, and rs5498, have been published as being associated with malaria [ICAM1 SNPs in association with clinical malaria phenotypes  in children was observed to confer risk , whereas the major polymorphism (G) in mothers was observed to confer risk . The relationship between genotype and phenotype here may be spurious, or potentially complex, requiring more specialized further studies. The literature supporting this association is mixed, however, more recent publications suggest there is no association between malaria and genotypes of rs5498 [Interestingly, the  malaria –40, 43,  malaria . Furtherf rs5498 , 45.Plasmodium spp. parasites to variant ICAM1 molecules (ICAM1Kilifi) further supports this observation [In these analyses, each polymorphism was analyzed using an additive model; a co-dominant model was also generated, but data not reported here. It is biologically plausible that, like many protective polymorphisms, the heterozygous genotypes have attained a skewed Hardy–Weinberg equilibrium in the population, and confer a phenotypic advantage against infection, which amounts to a small effect size . Existinervation , howeverervation .ABO gene provides evidence of risk factors predisposing to malarial infection in rs8176743 and rs8176746. The literature suggests that O blood phenotype is protective against malaria, though we could not correlate our findings with definitive blood typing. A possible mechanism of action is that infected O erythrocytes are more easily cleared than A or B, having reduced cytoadhesion compared to the A phenotype [ICAM1 and CD36, both of which are identified herein.Additionally, it is worth mentioning is that examination of the henotype . This isTaken in context, these findings complement the body of knowledge regarding malarial infection and host defence, while providing further information on candidate genes for future research. The list of genes analysed in this study , is not fully comprehensive, as it includes genes involved in the innate immune system and does not include niche genes such as those coding for sporozoite adhesion proteins, specific natural killer cell receptors, or expression of variable components of the adaptive immunity. However, a global interpretation of host risk genes here portrays a succinct and plausible narrative in that variations in major defence and signalling pathways involved in malarial infection are statistically associated as risk factors for clinical malaria outcomes and can, therefore, be targeted in future research for developing treatments and vaccines against malaria.TLR4 and related genes, ICAM1, IFNGR1, IL13, and IL6, as well as ABO, with the incidence of clinical malaria. In particular, the variations of T in the SNP rs5030719 of TLR4 conferred risk of clinical malaria compared to the G allele. This supports the current literature pertaining to the disease, as well as suggests that further research into the role of TLR4, as well as other related genes, in pathogenesis of clinical malaria may provide further insight into drug development and treatment of the disease.In conclusion, this study has utilized a comprehensive randomized control clinical trial to create a longitudinal dataset following maternal and children development to ascertain potential relationships between incidence of primary malaria cases with select candidate genes known to play a role in malarial pathogenesis. The results herein provide evidence for the association of Additional file 1: Table S1. Table of genes and single nucleotide polymorphismsanalysed in this study.Additional file 2: Table S2. Hardy–Weinberg equilibrium statistical values for child single nucleotide polymorphismsstudied herein. Only SNPs with a minor allele count of more than 5 were included in this analysis. The column for ‘chi-square’ values is calculated with 1 degree of freedom. The column for ‘p-value’ includes a continuity correction. Column ‘D’ denotes the disequilibrium coefficient for each SNP.Additional file 3: Table S3. Hardy–Weinberg equilibrium statistical values for maternal single nucleotide polymorphismsstudied herein. Only SNPs with a minor allele count of more than 5 were included in this analysis. The column for ‘chi-square’ values is calculated with 1 degree of freedom. The column for ‘p-value’ includes a continuity correction. Column ‘D’ denotes the disequilibrium coefficient for each SNP.Additional file 4: Table S4. Table of statistically significant single nucleotide polymorphismsand insertion/deletion eventscorrelated with episodes of anaemia or Plasmodium sp. Parasitaemia in child and/or mother. The column ‘rs_value’ lists the rs number associated with the SNP as defined in the NCBI dbSNP repository, ‘maternal/child’ lists the genome in which the SNP was identified, and ‘association’ lists the phenotype statistically associated with the SNP. Statistical columns respectively list the beta value for the association, standard error, Wald-value for hypothesis-testing, the p-value for the test, exponent of the beta value’), as well as the upper and lower values for the 95% confidence interval limit. The final columns, ‘ref’ and ‘alt’ denote the reference and alternate genotypes for SNPs and indels.


Plasmodium falciparum) are common in school-aged children and represent a disease transmission reservoir as they are potentially infectious to mosquitoes. To detect and treat such infections, convenient, rapid and reliable diagnostic tools are needed. In this study, malaria rapid diagnostic tests (mRDT), light microscopy (LM) and quantitative polymerase chain reaction (qPCR) were used to evaluate their performance detecting asymptomatic malaria infections that are infectious to mosquitoes.Asymptomatic malaria infections (Plasmodium spp. infections using mRDT (SD BIOLINE), LM and qPCR. In addition, gametocytes were detected using reverse transcription quantitative polymerase chain reaction (RT-qPCR) for all qPCR-positive children. Venous blood from all P. falciparum positive children was fed to female Anopheles gambiae sensu stricto mosquitoes via direct membrane feeding assays (DMFAs) after serum replacement. Mosquitoes were dissected for oocyst infections on day 8 post-infection.One hundred seventy asymptomatic school-aged children (6–14 years old) from the Bagamoyo district in Tanzania were screened for P. falciparum prevalence in study participants was 31.7% by qPCR, 18.2% by mRDT and 9.4% by LM. Approximately one-third (31.2%) of asymptomatic malaria infections were infectious to mosquitoes in DMFAs. In total, 297 infected mosquitoes were recorded after dissections, from which 94.9% (282/297) were derived from infections detected by mRDT and 5.1% (15/297) from subpatent mRDT infections.The The mRDT can be used reliably to detect children carrying gametocyte densities sufficient to infect high numbers of mosquitoes. Subpatent mRDT infections contributed marginally to the pool of oocyts-infected mosquitoes.The online version contains supplementary material available at 10.1186/s13071-023-05761-w. Plasmodium falciparum opens new avenues to tackle malaria infections that are asymptomatic and infectious to mosquitoes [Plasmodium transmission due to persistent asymptomatic infections [Plasmodium infections and reduce the transmission reservoir [Better knowledge on treatment strategies targeting the infectious reservoir of squitoes . Asymptosquitoes  and are squitoes , 4 and hsquitoes . Some prsquitoes , 7 and cfections , 8. A maeservoir , 10, espeservoir .In previous studies, mRDTs have been used for detection of asymptomatic malaria infections. For example, in community studies, a limited reduction in malaria transmission was reported from Zambia  while inLow-density asymptomatic infections—including those with gametocyte densities potentially infectious to mosquitoes—are often undetected by LM and mRDT, the standard diagnostic tools in malaria diagnostics. When compared to more sensitive molecular tools such as qPCR, LM and mRDT have repeatedly shown inferior performance –18. In gHere, we investigated the diagnostic performance of a commercially available mRDT (SD BIOLINE) compared with LM and qPCR for detecting infectious and asymptomatic malaria infections among school-aged children. To confirm onward transmission to mosquitoes DMFAs were performed. Our aim was to investigate whether parasite densities subpatent to mRDTs are infectious to mosquitoes in DMFAs. Using qPCR and RT-qPCR, we measured the infectiousness of microscopic and submicroscopic gametocyte densities from humans to mosquitoes.Plasmodium infection were enrolled for blood-drawing for DMFAs and then treated with artemether-lumefantrine (ALU). The treatment was done within 24 h of diagnosis according to national guidelines in Tanzania [This cross-sectional study was conducted in two primary schools of Buma and Yombo located in the Bagamoyo district in coastal Tanzania. After the long rainy season that lasts from March to June, we recruited 170 asymptomatic school-aged children over a period of 10 weeks between June and the end of August 2020. Children aged 6–14 years were eligible for enrolment when written informed consent from their caregivers and assent from the children was obtained. All children screened positive for Tanzania . An overAll participants were finger pricked using a safety lancet  to obtain a capillary blood sample into an EDTA containing collection tube . Then, 180 µL whole blood was transferred into Eppendorf tubes containing 540 µL of 1 × DNA/RNA Shield™  for nucleic acid preservation until extraction of DNA (same day of preservation).Plasmodium using mRDT , Standard Diagnostic, South Korea) was executed according to the manufacturer’s instructions. The mRDT detects the P. falciparum-specific histidine-rich protein 2 (HRPII) antigen targeting the parasites asexual blood stages and Plasmodium species in general via the parasite lactate dehydrogenase (pLDH), including sexual stages (gametocytes). One drop (~ 50 µL) of capillary blood was applied to the test and run with four drops of a delivered standard buffer. The test was kept on a flat surface while running and read by the laboratory technician after 15 min. The result was confirmed and recorded by a second member of the field team trained on reading the mRDT.Examination of the presence of Plasmodium parasites under a light microscope . Slides were considered negative if no Plasmodium parasites were detected after examination of 100 consecutive fields. After detection of the first parasites, asexual parasites (ring stages) were counted per 200 white blood cells (WBCs) and gametocytes were counted per 500 WBCs.Thick smears were prepared using capillary blood from the finger prick. Dried thick smears were stained for 45 min with 10% Giemsa stain and examined for the presence of Plasmodium infections was manually extracted from 180 µL whole blood preserved as described above using the Quick-DNA™ Miniprep kit (Zymo Research) according to the manufacturer’s instructions. DNA was eluted in 50 µL elution buffer and stored at − 20 °C.The DNA for molecular diagnosis and quantification of Plasmodium falciparum infections were detected and quantified using the PlasQ qPCR assay [P. falciparum quantification [P. falciparum NF54 DNA as positive control (PC).CR assay , 24. To fication  was usedP. falciparum co-infections with other relevant parasite species in the Bagamoyo district [Plasmodium-positive samples were subjected to further analysis using the PlasID qPCR assay as previously described [Plasmodium species were included. The thermal profile used for the assay was as follows: 5 min at 95 °C; 45 cycles of 15 s at 95 °C and 60 s at 55 °C.To investigate district , 26, allescribed . SamplesSamples for ribonucleic acid (RNA) extraction were taken from all qPCR-positive (PlasQ assay) children and 150 µL of whole blood was transferred into an Eppendorf tube containing 450 µL 1 × DNA/RNA Shield™. The tubes were mixed by inversion, spun down for 30 s and incubated for 60 min at room temperature before subsequent storage at − 20 °C until further use. RNA was manually extracted using the Quick-RNA™ MiniPrep Plus kit (Zymo Research). The protocol was adapted as per manufacturer’s suggestions: (i) after thawing of frozen samples, 100 µL of 1 × DNA/RNA Shield™ was added and vortexed for 60 s at full speed; (ii) Proteinase K digestion was extended to 60 min at room temperature; (iii) after Proteinase K digestion, the samples were centrifuged for 5 min at 16,000 × rcf to pellet all debris of red blood cells (RBCs). On-column DNase I digestion of gDNA was performed on all samples according to the manufacturer’s instructions . RNA wasFemale gametocytes were detected using a RT-qPCR assay designed by Wampfler et al. , 30. RevAnopheles gambiae sensu stricto (Ifakara strain) colony, which is reared at IHI, Bagamoyo Branch Insectary, Kingani. The colony is maintained under standard conditions at a 12:12 h dark:light cycle, 70 ± 20% humidity and 27 ± 2 °C. Larvae are fed on TetraMin® fish flakes  and reared at a density of 200 larvae  per 1 L deionized water. Adult mosquitoes are kept in 30 × 30 × 30 cm containers and allowed to feed on 10% sucrose solution. Adult mosquitoes are routinely checked for microsporidia infections [We used the fections , 32. CagP. falciparum-positive, asymptomatic school-aged children was drawn for DMFAs in lithium-heparin-coated vacutainers. Whole blood was transferred in pre-warmed 1.5 mL Eppendorf tubes centrifuged for 3 min at 300 rcf and 37 °C [Unfed female mosquitoes  were sugar starved overnight with access to water only. The next morning, a batch of 50 female mosquitoes was transferred into paper cups. For each participant, six cups were prepared. Blood from nd 37 °C . Autolognd 37 °C  (37 °C).nd 37 °C . After tData was entered in Excel  and analysed using the statistical software R and the R studio (Version 1.3.1093) interface. Sensitivity of each diagnostic test was manually calculated using 2 × 2 contingency tables. Proportions of positive and negative events of diagnostic tests including 95% confidence intervals (CIs) were calculated by one-sample z-tests. Positive predictive values of diagnostic tests in predicting infected mosquitoes deriving from infectious asymptomatic individuals were calculated as percentages of infected mosquitoes of the respective diagnostic tool, divided by all infected mosquitoes detected with qPCR.N = 80) and Yombo (N = 90) primary schools. Mean age among study participants was 10.3 years and participants were 64% female (N = 109) and 36% male (N = 61).A total of 170 children participated in the study from Buma  infection. No mixed infections were detected. Asexual blood-stage densities of P. falciparum molecularly quantified by qPCR ranged from 0.1 parasites/µL to 275,000 parasites/µL with a median parasite density of 6 parasites/µL. About 18.2% of study participants were mRDT positive . Of 170, 54 were qPCR-positive for P. falciparum-positive children, six were not enrolled for DMFAs and were treated directly with ALU. Reasons for exclusion were developing symptoms (n = 2), withdrawn from the study by caregiver (n = 1), fear of needle pricking for blood drawing (n = 2) and insufficient mosquito numbers for DMFAs (n = 1). The remaining 48 qPCR-positive children were enrolled for DMFAs. From those, 15  All infected mosquitoes derived from infectious participants detected by qPCR and RT-qPCR. Table B. Nine pThe aim of this study was to compare the performance of standard diagnostic tools (mRDT and LM) to a sensitive molecular tool (qPCR) in detecting asymptomatic malaria infections that are infectious to mosquitoes. To our knowledge, this is the first study that specifically assesses the performance of an mRDT including the onward transmission of detected infections to mosquitoes using DMFAs.P. falciparum infections that are infectious to mosquitoes are detected by standard diagnostic tools is consistent with evidence from other African countries [We observed that nearly one-third (31.2%, 15/48, Fig. ountries , 36. WheFurthermore, we report that 85% of all infected mosquitoes derived from only four children; all of them were detected by mRDT and three were LM-positive for gametocytes Fig. C. Our fiWe have demonstrated that mRDTs identified a slightly higher number of the qPCR-positive infections compared to LM. While LM performance may be further improved by including a second microscopist, the success of the mRDT was encouraging and indicates its potential to be used alone for detection of infectious asymptomatic individuals. The advantage of using mRDTs over LM is that it is convenient and does not need highly skilled experts to interpret the results. Our findings on sensitivity and specificity concur with a previous study that evaluated the same mRDT product in Tanzania . Also siThe reported malaria prevalence places the Bagamoyo district as a moderate transmission area , similarIn our study, a small proportion of children was LM positive for gametocytes, but when applying molecular diagnostics, we found that nearly every fifth child carried female gametocytes as detected by RT-qPCR. These results were not surprising and comparable to gametocyte prevalence detected by LM and RT-qPCR in previous studies from Tanzania , 43 and Measuring the transmissibility of malaria infections using DMFAs has several limitations . Here, wCommercially available mRDTs may be a suitable and convenient tool to identify asymptomatic malaria infections that are infectious to mosquitoes. Although mRDTs failed to detect a substantial number of asymptomatic malaria infections compared to the total infections detected by qPCR, the mRDT detected infections that accounted for 95% of all oocyst-infected mosquitoes after DMFAs. Many low-density infections that are subpatent to mRDTs could become infectious to mosquitoes at a later time point. Therefore, repeated testing with mRDTs and treatment may be required if implemented as an intervention to reduce the reservoir of transmission and numbers of infectious mosquitoes in a community efficiently . FinallyAdditional file 1: Oligos used in this study.Additional file 2: Complete data set of the present study.


Malaria during pregnancy is a priority area for malaria research and control as pregnant women represent a high risk group for severe malaria, and the presentation of malaria during pregnancy varies according to the level of transmission in the area; so the aim of this study is to determine the prevalence rates of malaria parasite among pregnant women attending to Saudi Kassala Teaching hospital in Kassala state, 2022.  A cross-sectional study was carried out in Saudi Kassala Teaching hospital in Kassala State. This study involved one hundred and eighty-five blood samples collected from pregnant women who was then examined by using blood films and ICT for malaria, and the data were collected by a structured questionnaire and analyzed using SPSS version 21. P value = 0.483). The prevalence of malaria in rural residency was 2.2%, and this was significantly more common than the urban residency . When compared across the gestational trimesters, there was no significant difference between them . The number of gravidity is not related to malaria infection . The presence of symptom compliant of malaria during pregnancy does not suggest the presence of malaria . No difference was found between the different educational levels with respect to the prevalence of malaria . The result showed that there was 1 (0.5%) negative result in ICT which was positive in blood film for malaria (BFFM) and there were 3 (1.6%) positive malaria parasites by both methods in all 185 samples with statistically insignificant differences (P = 0.703).  The prevalence of malaria among pregnant women was 2.2% (95% CI: 0.006–0.054). There was no significant difference among the different age groups with respect to the prevalence of malaria (Plasmodium falciparum was only species detected in this study. Malaria among pregnant women was more prevalent in rural areas. However, other factors such as age, gestational age, gravidity, and educational level do not affect the prevalence of malaria in pregnant women. The presence of symptomatic compliant of malaria during pregnancy does not suggest the presence of malaria. The use of ICT or BFFM has similar diagnostic outcome for malaria in pregnancy. Plasmodium species, and P. falciparum is more common in Africa. According to WHO, Sudan carries the heaviest burden accounting for 61% of deaths in the Eastern Mediterranean [Plasmodium parasites, Plasmodium falciparum is responsible for the highest disease burden [Plasmodium falciparum [Plasmodium falciparum which cause high levels of parasitemia, hypoglycemia, acute pulmonary edema, fetal distress, premature labor, spontaneous abortions, and still births [Malaria is caused by erranean . Of all e burden . The othe burden . An intee burden . Other re burden . The maie burden . In prege burden . In sub-e burden . This islciparum . In arealciparum . Pregnanl births . The preThe study was a descriptive cross-sectional hospital-based study, conducted from February to September 2022. It was conducted in Saudi Kassala Teaching hospital in Kassala state among 185 pregnant women attending the hospital for either monthly follow-up visit or delivery. Kassala state is located in the east of the Sudan. It has a total population of 1.5 million .This state is served by Saudi Kassala teaching hospital, which is the only hospital that provides antenatal and delivery service to the state population. In 2022, the hospital had 7498 spontaneous vaginal deliveries, 18 assisted vaginal deliveries through ventouse suction, and 5,586 cesarean section .z is the normal standard deviation (1.96), p is the frequency of occurrence of an event, q is the frequency of nonoccurrence of an event, and d is the degree of precision (0.05) [Sample size was calculated using the following formula:n (0.05) .So, when the margin of error is 5%, confidence level is 95%, and prevalence is 13.7% .(2)n=1.9The calculated sample is 182; however, a sample of 185 was taken for more accuracy and precision.Inclusion criteria included pregnant women attending Saudi Kassala Teaching hospital from February to September 2022.Exclusion criteria included pregnant women referred from outside of the state of Kassala and pregnant women who were diagnosed with malaria by laboratories other than the Saudi Kassala Teaching hospital laboratory.A 5 ml of blood sample was collected from pregnant women in EDTA container. Thin and thick microscopic examination and immunochromatographic test (ICT) were used for confirming the diagnosis.Thick and thin blood films were air-dried. Methanol was used to fix the thin films. 10% Giemsa solution was used to stain the films, and the stain was allowed for 10 minutes, then washed with clean water, and left to air-dry. 100x magnification lenses under oil immersion were used to view the films .Plasmodium falciparum is coated with monoclonal antibodies against histidine-rich protein 2 while the second line for Plasmodium vivax is coated with monoclonal antibodies specific to lactate dehydrogenase [This test has a membrane strip coated with monoclonal antibodies. The first test line for rogenase .µl of anticoagulated blood using a micropipette, and two drops of buffer was added in port B using plastic dropper. The results were interpreted after 20 minutes [At room temperature, port A in the test kit was filled with 5  minutes .After estimating the sample, a questionnaire was designed to collect the demographic data as well as data related to the gestational age, gravidity, and malaria symptoms during pregnancy.The data were analyzed using frequencies and cross-tabulation using chi-square in Statistical Package for Social Science (SPSS) computer program version 21.All the examination protocols were approved by the ethical committees of both the faculty of medicine and health sciences at the University of Kassala and Saudi Kassala Teaching hospital in Kassala State. The informed written consent was taken from each pregnant women participated in this study.Pregnant women who are less than 16 years of age were married, and as a result, they were considered emancipated, and the informed written consent was taken accordingly.Informed written consent from illiterate pregnant women was taken with the participation of the husband as a legal guardian.Of the 185 women approached, the data were successfully collected and no refusal was noted.All the methods were carried out in accordance with local regulations and guidelines.Plasmodium falciparum  were infected with malaria parasite, and all of them were infected by lciparum .P value = 0.483  were positive in the age group 15–25 years and 1 (1.1%) in the age group of 26–35 years. There was no significant difference among the different age groups with respect to the prevalence of malaria at  = 0.483 .P = 0.021  compared with 0 (0%) positive cases in the urban residency, and these differences in rates were significant at  = 0.021 .P = 0.362  were positive for malaria parasite, and out of the 39 who had secondary educational level, 1 (2.6%) was positive for malaria parasite, while the other educational groups level was negative for malaria parasite with statistically insignificant differences in rates at  = 0.362 .P = 0.518  positive for malaria, with statistically insignificant differences rate at  = 0.518 .P = 0.737 , 1 (3.0%), 2 (3.0%), and 0 (0.0%) positive case among primigravidity, second gravidity, multigravidity, and grand multigravidity, respectively, with statistically insignificant differences in rate at  = 0.737 .Out of the 39 pregnant women complaining of malaria symptoms which include fever, nausea, vomiting, anorexia, chills, headache, and myalgia , only 2 P = 0.703 (The result showed that there was 1 (0.5%) negative result in ICT which was positive in BFFM and there were 3 (1.6%) positive malaria parasites by both methods in all 185 samples, with statistically insignificant differences at  = 0.703 .The study showed that the prevalence of malaria among pregnant women was 2.2% (95% CI: 0.6–5.4). Meanwhile, the department of statistics at Saudi Kassala Teaching hospital recorded a prevalence of 1.79% in 2020 and 1.57% in 2011. This is relatively similar to the results of our study, and these indicate that malaria among pregnant women was relatively stable throughout the last 3 years .Plasmodium falciparum was the only species of malaria parasite detected, and this was similar to that of a study done in Al Jabalian Locality, White Nile state, Sudan, which has similar result [Plasmodium falciparum is the predominate species in Africa [According to our study, r result . This aln Africa .We found that the highest prevalence of malaria in pregnancy was 3.5% recorded among age group of 15–25 years. However, there was no significant association between the prevalence of malaria and the age group of pregnant women, and this finding is similar to another study done in eastern Sudan . This siMalarial infection was significantly higher in women from rural areas compared to woman from urban areas; this finding was similar to the findings reported by Omer and colleagues . This hiEven though malaria prevalence is relatively higher in the noneducated group (4.3%), there is no significant association between the education level and malaria infection, and this finding agrees with the finding reported by Suliman and colleagues .There is no association between the gestational age and malaria, and this finding is similar to that of a study done by Tahita and colleagues .This study showed no significant association between malaria and gravidity, and this finding was similar to that of another study done in eastern Sudan by Adam and colleagues .Our study shows that symptoms of malaria were not discriminative for malaria infections . This is similar to the findings of the study done by Tahita and colleagues . The difP value = 0.751). This was similar to the results obtained by Latha and colleagues where rapid diagnostic test was found equally sensitive to microscopy [When the blood films and ICTs were compared for the diagnosis of malaria parasite, the difference between the two modalities was not statistically significant (croscopy .Plasmodium falciparum was the only species detected in this study.Malaria among pregnant women was more prevalent in rural areas. However, other factors such as age, gestational age, gravidity, and educational level do not affect the prevalence of malaria in pregnant women.The presence of symptomatic compliant of malaria during pregnancy does not suggest the presence of malaria.The use of ICT or BFFM has similar diagnostic outcome for malaria in pregnancy.


Malaria is a global public health concern, mainly occurring in sub-Saharan Africa. Children infected with malaria are more likely to develop severe disease, which can be fatal. During COVID-19 in 2020, diagnosing and treating malaria became difficult. We analyzed the clinical characteristics and laboratory indicators of children with severe malaria in Benin to provide important information for designing effective prevention and treatment strategies to manage pediatric cases.  Clinical characteristics of pediatric patients with severe malaria admitted to two hospitals in Benin  were collected from January to December 2020. Patients were grouped according to age , and clinical and laboratory indicators were compared. The incidences of severe pediatric malaria in both hospitals in 2020 were calculated. Inclusion, exclusion, and blood transfusion criteria were identified. p < 0.05), and the leukocyte count was significantly lower (p < 0.01) than in groups A and B. Parasitemia was significantly higher in group C than in group A (p < 0.01). Twelve deaths occurred.  We analyzed 236 pediatric cases. The main clinical symptoms among all patients were severe anemia, vomiting, prostration, poor appetite, dysphoria, and dyspnea. Over 50% of patients in group A experienced vomiting and severe anemia. Most patients in group B had severe anemia and prostration. Delirium affected significantly more patients in group C than in groups A and B. In group C, the hemoglobin and hematocrit levels were significantly higher ( Severe pediatric malaria is seasonal in Benin. The situation in children under 5 years old is poor. The main problems are severe disease conditions and high fatality rates. Effective approaches such as prevention and early and appropriate treatment are necessary to reduce the malaria burden in pediatric patients. Plasmodium falciparum. The percentage of total malaria deaths among children aged under 5 years decreased from 87% in 2000 to 76% in 2019, but increased slightly to 77% in 2020 [Malaria is a major global public health concern . Accordi in 2020 . A WHO rPlasmodium species that affect humans are Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Plasmodium ovale, and Plasmodium knowlesi. P. falciparum is the main causative agent of malaria in endemic areas. Severe malaria is primarily due to P. falciparum infections, followed by P. vivax and P. knowlesi infections [P. falciparum present in Africa [P. falciparum infection every 2 minutes [The five malarial fections , 5. The n Africa . The lat minutes .P. falciparum is the primary species; P. vivax, P. malariae, and P. ovale are less common. In 2019, the incidences of malaria, malaria-related mortality, severe malaria, and severe pediatric malaria were 21.2%, 1.3‰, 1.7%, and 4.34%, respectively.Malaria is a major public health concern in Benin. In 2019, malaria cases constituted >38% of outpatients and >60% of inpatients nationwide. In March 2020, the WHO issued guidelines for responding to malaria during the COVID-19 pandemic . These gIn 2020, our team conducted medical surveys of two hospitals in Benin. We collected and analyzed clinical data on children under 5 years old with confirmed malaria who were admitted to these hospitals from January to December 2020. This study was conducted to provide basic clinical information and recommendations for treating severe pediatric malaria in Benin.The two hospitals in Benin involved in this study were the Central Hospital of Lokossa and the Regional Hospital of Natitingou. The Central Hospital of Lokossa is in northwest Mono-Couffo Province and is 106 km from Cotonou, the capital of the Republic of Benin. The Regional Hospital of Natitingou is in Accra Province, a hilly area in northern Benin. Both hospitals are located approximately 650 kilometers apart. Benin, a narrow north-south key-shaped strip of land in West Africa, is situated between the Equator and the Tropic of Cancer. The country is divided into four main regions from south to north. The coastal plain region has a tropical rainforest climate, the central and northern regions have a savannah climate, and the south has a Guinea-type climate.Lokossa in southwest Benin is a coastal area with elongated coastlines and extensive marshes. The city mainly consists of low-lying sandy coastal plains that stretch towards the Atlantic Ocean, along with marshes, lagoons, and lakes. Southern regions of Benin have two rainy periods from March to July and September to November, while the northern regions have a single rainy season from May to September. Natitingou, a town belonging to the Department of Atakora in northwestern Benin, lies in a semi-valley enclosed by two mountain ridges. The climate is drier in Natitingou than in the south, especially during the Harmattan season in December and January when humidity drops as low as 10% and night temperatures can reach as low as 17°C. Consequently, the climatic conditions in Lokossa are more favorable for mosquito breeding.Clinical characteristics and indicators were recorded for the enrolled children with severe malaria, who were admitted to the two hospitals from 1 January 2020 to 31 December 2020. The patients were grouped according to age: group A , group B , and group C . The patients' legal guardians provided informed written consent. The principles outlined in the Declaration of Helsinki were followed during the study.Inclusion criteria were pediatric inpatients conformed to the diagnostic criteria for severe malaria, patients had consistent treatment plans, and cases included consent from the guardian and hospital director. Exclusion criteria were pediatric patients who did not conform to the diagnostic criteria for severe malaria, patients with different treatment options, patients whose guardians did not consent to their enrolling in the study, patients who quit before completing the treatment course, and patients with congenital diseases.P. falciparum malaria with impaired consciousness (a Blantyre coma score of <3 of 5 in children); (2) prostration (inability to sit unassisted due to extreme weakness), multiple convulsions , acidosis (plasma bicarbonate concentration <15 mM or intravenous plasma lactate level >5 mM), hypoglycemia (whole blood or plasma glucose concentration <2.2 mM or 40 mg/dL), severe anemia (hemoglobin <5 g/dL or hematocrit <15%), renal impairment (elevated plasma creatinine >265 μM or blood urea >20 mM), jaundice , pulmonary edema (tachypnea (respiratory rate >30/min)), dyspnea and hypoxia (oxygen saturation <92%), compensated shock (capillary refill ≥3 s without hypotension after rehydration), decompensated shock , or hyperparasitemia (parasitemia >10% without other severe signs).The latest WHO guidelines for malaria treatment (third edition) and the Practical Manual for the Management of Severe Malaria (third edition) define severe malaria as follows: (1) positive microscopic examination or rapid diagnostic test for Artemisinin was injected at 3 mg/kg, three times daily for patients weighing ≥20 kg and at 2.4 mg/kg, three times daily for patients weighing <20 kg. Oral artemether was used twice daily at 12 h intervals for 3 days. Patients weighing 5–14 kg received 120 mg artemether; those weighing 15–24 kg received 240 mg artemether; those weighing 25–34 kg received 360 mg artemether; and those weighing >34 kg received 480 mg artemether.Notably, because of Benin's hygiene conditions and medical level, treatment programs for severe malaria are not fully implemented according to global treatment programs for severe malaria.Both hospitals adhered to the WHO's transfusion criteria . In highPlasmodium in peripheral blood films [P. falciparum-associated malaria [µL or >5%), erythrocyte volume fraction <15%, hemoglobin concentration <5 g/dL, blood glucose <2.2 mmol/L (<40 mg/dL), blood urea >60 mg/dL, and serum creatinine >265 µmol/L (>3.0 mg/dL).Cured was defined as the absence of clinical symptoms and no od films . The maj malaria  used by P. falciparum was detected in the thick film of patients' peripheral blood under light microscopy after fixation and staining with Giemsa. The parasite load was calculated using the formula: number of parasites/number of white blood cells × exact white blood cell count. If the exact count was unknown, the average normal white blood cell count in Benin (6000 GB/UL) was used for the calculation [9/L), red blood cell count , platelet count , C-reactive protein level , and Plasmodium count .culation . The folt-tests. Nonnormally distributed data are expressed as medians with interquartile range and compared using nonparametric tests. Chi-square tests were used to compare categorical data. The statistical significance threshold was set at p < 0.05.Clinical characteristics and indicators were compared among groups. Statistical analysis was performed using SPSS software . Normally distributed data are expressed as means ± standard deviation and compared via P. falciparum-induced malaria were analyzed. Twelve deaths occurred, yielding a fatality rate of 5.10%. We divided the 236 patients into groups A, B, and C according to age, as described in the Methods section, and compared the clinical characteristics and indicators among them  with severe ong them .Benin has only rainy and dry seasons. Data from the two hospitals show that in 2020, the incidence of severe malaria in children was higher during the rainy seasons, especially from June to November, with the highest incidence in September .The main clinical symptoms among all patients were severe anemia, vomiting, prostration, poor appetite, dysphoria, and dyspnea. Some clinical symptoms differed among groups. More than 50% of patients in group A experienced vomiting and severe anemia, while most of group B had severe anemia and prostration. Delirium affected significantly more patients in group C than in groups A and B .p < 0.05), and the leukocyte count was significantly lower (p < 0.01) than in groups A and B (Plasmodium count was significantly higher in group C than in group A (p < 0.01). The remaining laboratory indicators and the average hospital stay lengths did not significantly differ among the groups .In group C, the hemoglobin levels, hematocrit levels, and red blood cell counts were significantly higher  in their erythrocytes, and the low levels of iron and para-amino benzoic acid in breast milk , 26. MalP. falciparum-induced malaria in areas of high transmission. Severe anemia develops rapidly after infections with high parasitic density and occurs earlier and more significantly. In these cases, acute destruction of parasitized red blood cells, which causes the anemia, requires careful monitoring during treatment [Severe anemia is the leading cause of death in children with malaria and is a common presenting feature of reatment . In our reatment , 28.9/L in some of our patients, which is consistent with previous findings [Plasmodium-containing erythrocytes in the spleen, involvement of platelets in repairing Plasmodium-damaged vascular endothelial function, excessive platelet aggregation and damage caused by Plasmodium antigen-mediated immune mechanisms, and disrupted maturation of megakaryocytes in the bone marrow [9/L) [Plasmodium removal, platelet counts can return to normal. In this study, platelet counts did not significantly differ among groups, indicating that platelet counts do not substantially change with age.Platelet counts are a critical factor in diagnosing malaria . Platelefindings . Some poe marrow . Therefoow [9/L) , 33. AftLeukocyte and neutrophil counts can increase during acute paroxysm, remain normal after the paroxysm, and decrease after multiple paroxysms . In thisPlasmodium infection, red blood cells experience endovascular adhesion damage. Second, Plasmodium parasites cause microvascular lesions and pathological reactions in the brain, lungs, and other organs. Third, Plasmodium parasites cause widespread secretion of C-reactive protein into blood [Several factors might contribute to changes in C-reactive protein levels in patients with malaria. First, after to blood . In thisThis study had some limitations. First, because of financial limitations, patients were generally hospitalized for 2–4 days. After patients received blood transfusions and antimalarial treatment, their guardians often requested discharge if the patient's condition was improved. If financial conditions allowed, the children were discharged with instructions to continue oral medication at home. If the financial conditions were poor, the children were only observed after returning home. Incomplete laboratory registration books were also a limitation.In West African nations such as Benin, where the malaria incidence and mortality are high, comprehensive treatment programs are needed. Currently, West African nations with high malaria incidences have weak monitoring systems. Malaria must be eliminated as well as diagnosed and treated. The following factors should be considered: increasing investment in domestic and international resources to enable assessing malaria patients with uniform criteria; investing public health resources where the population has the greatest need and the intervention will be most effective; strengthening the training and technical guidance for local healthcare staff and developing appropriate local malaria elimination measures consistent with scientific assessment of the epidemic scope and transmission characteristics; and providing practical control packages .Severe malaria remains problematic in children under 5 years old. The main problems are disease conditions and high fatality rates. Effective approaches such as infection prevention and early and appropriate treatment are necessary to reduce the malarial burden in endemic settings. The international community should continue to pay attention to this life-threatening disease and assist special populations, such as young children and pregnant women, in diagnosing and treating malaria in sub-Saharan Africa.


Monthly height and weight, and time of incident malaria episodes were obtained; 89 children who received DP contributed piperaquine (PQ) concentrations. Malaria hazard was modeled using parametric survival analysis adjusted for repeated events, and height and weight were modeled using a Brody growth model. Among 579 children, stunting (height‐for‐age z‐score [ZHA] < −2) was associated with a 17% increased malaria hazard  compared with children with a ZHA of zero. DP was associated with a 35% lower malaria hazard , compared to no chemoprevention. After accounting for PQ levels, stunted children who received DP had 2.1 times the hazard of malaria (HR [95% CI] 2.1 [1.6–3.0]) compared with children with a ZHA of zero who received DP. Each additional malaria episode was associated with a 0.4% reduced growth rate for height. Better dosing regimens are needed to optimize malaria prevention in malnourished populations, but, importantly, malaria chemoprevention may reduce the burden of malnutrition in early childhood.African children are at risk of malaria and malnutrition. We quantified relationships between malaria and malnutrition among young Ugandan children in a high malaria transmission region. Data were used from a randomized controlled trial where Ugandan HIV‐unexposed ( WHAT IS THE CURRENT KNOWLEDGE ON THE TOPIC?The greatest burdens of malnutrition and malaria overlap in sub‐Saharan Africa, but little is known about the relationships among malnutrition, malaria, and the efficacy of malaria chemoprevention.WHAT QUESTION DID THIS STUDY ADDRESS?We used parametric survival and growth models to answer whether malnutrition is associated with an increased hazard of malaria and whether malaria is associated with decreased childhood growth.WHAT DOES THIS STUDY ADD TO OUR KNOWLEDGE?We show that malnutrition severity is associated with an increased hazard of incident malaria even when effective malaria chemoprevention is administered. Additionally, our findings indicate that each additional episode of malaria is associated with a reduced growth rate for height.HOW MIGHT THIS CHANGE DRUG DISCOVERY, DEVELOPMENT, AND/OR THERAPEUTICS?This study highlights the importance of including malnourished children in clinical trials. Stunted children have a higher risk of malaria regardless of chemoprevention regimen, suggesting that more work is needed to optimize chemoprevention regimens for this population. Future clinical trials should (1) be powered to evaluate malnutrition as a covariate and (2) incorporate evaluation of nonmalarial outcomes, including malnutrition/childhood growth.Malaria and malnutrition are significant causes of morbidity and mortality in young children in sub‐Saharan Africa. Children less than 5 years of age accounted for 77% of malaria deaths globally in 2020,Chronic malnutrition, as measured by low height‐for‐age z‐score (ZHA), in young African children has been associated with increased risk of malaria, severe malaria, and anemia.It is also likely that malaria exacerbates malnutrition. Use of long‐lasting insecticide treated bed nets (LLINs) for malaria prevention led to greater weight and height gains in children when compared to no LLIN use.We identified and quantified risk factors for malaria and malnutrition in a large cohort of Ugandan children who were followed from 6 months to 3 years of age as part of two randomized controlled trials of three malaria chemoprevention regimens.Data from two randomized controlled malaria chemoprevention trials conducted concurrently in the same clinic in Tororo, Uganda, from June 2010 through September 2013 were used.HIV‐unexposed children began chemoprevention at 6 months of age and, because HIV‐exposed participants received daily trimethoprim‐sulfamethoxazole (TS) while breastfeeding per national guidelines, they began chemoprevention at 6 months of age or upon breastfeeding cessation, whichever was later. All participants received chemoprevention until 24 months of age and were followed off chemoprevention until 36 months of age. Children were randomized in equal numbers to no chemoprevention, a once monthly dose of sulfadoxine‐pyrimethamine (SP), a daily dose of TS, or a monthly treatment course (daily dosing for 3 consecutive days) of dihydroartemisinin‐piperaquine (DP). Each regimen was administered according to weight‐bands specified by the manufacturer at the time of the study Table .18, 1Children received all medical care at a dedicated study clinic to ensure accurate diagnosis of incident malaria. Children returned monthly for weight and height measurements and the parent/guardian was asked if the child had breast fed in the last 7 days. At any visit, if a child had a fever (>37.5°C) or a history of fever within the last 24 h, a thick blood smear was taken to evaluate for malaria. If the blood smear was positive, the child was diagnosed with and treated for malaria. The time of incident malaria was defined as the day of a visit where malaria was diagnosed which occurred greater than 14 days after a prior episode.Monthly age, weight, and height measurements were used to calculate weight‐for‐age z‐score (WAZ), ZHA, and weight‐for‐height z‐score (WHZ). Z‐scores were calculated using the World Health Organization's (WHO) Anthro macro.A subset of HIV‐unexposed children who received DP underwent sampling of venous plasma to determine piperaquine (PQ) concentrations.p value of less than 0.05 for forward inclusion and a p value of less than 0.01 for backward elimination using linear and exponential functions.A parametric survival model, adjusted for repeated events, was developed to estimate the hazard of incident malaria from 6 to 24 months of age, during chemoprevention. The Laplace estimation method in NONMEM, version 7.4.2 was used.Model selection was guided by change in objective function value (OFV) between nested models, parameter estimates, relative standard error (RSE), and Kaplan–Meier visual predictive check.n = 1000).Hazard ratios (HRs) with 95% confidence intervals (CIs) were calculated using the final parameter estimates and covariance matrices. Percent reductions in incident malaria hazard were defined as 1 – the HR. For the final models, 95% CIs of parameter estimates were determined by bootstrap .Growth models were developed to predict height and weight from 6 months to 3 years of age. Monthly height, weight, and age measurements were described using a Brody growth model (Equations listed in n = 1000) to predict the total number of malaria episodes in stunted (ZHA = −2.1) and non‐stunted (ZHA = 0) children. For simulations, children were assumed to be HIV‐unexposed and in the low or middle socioeconomic category. For the DP arm, we conducted 1000 simulations of the above patients, but stratified into groups with PQ concentrations consistently greater than 10.3 ng/ml or less than or equal to 10.3 ng/ml.The final survival models were used for simulations  children at 3 years of age, stratified by cumulative malaria episodes. Four datasets comprised of baseline WHZ, ZHA, sex, HIV exposure status, and time varying cumulative malaria episodes  from 6 months to 3 years of age were simulated 100 times. Predicted ZHA was calculated using the WHO Anthro macro.Among the 579 children included in the analysis, 186 (32.1%) were HIV‐exposed. HIV‐exposed children were older when they began study drugs compared with HIV‐unexposed children (median age: 10 months vs. 6 months) and had lower median Z‐scores Table . In totan = 283, 56.6%) or at monthly routine visits  from 89 HIV‐unexposed children who received DP. The median PQ concentration was 5.6 ng/ml (range: LLOQ – 316 ng/ml). Children with a preceding PQ concentration of greater than 10.3 ng/ml had a lower cumulative hazard of repeated malaria events compared to when the preceding PQ concentration was less than or equal to 10.3 ng/ml  of monthly study visits. Over 88% of measurements were below the WHO defined mean ZHA and 50% met the criteria for stunting (ZHA < ‐2) at least once. Children who never developed malaria between 6 months and 3 years of age trended towards being taller than those who had malaria greater than or equal to one time  and provided similar statistical significance and effect size. However, uncertainty in the covariate parameter estimate was higher (% relative standard error [%RSE] = 77.4) when WAZ was included compared to ZHA (%RSE = 17.8) and ZHA was included in the final model. Age, WHZ, weight, and sex were not associated with malaria hazard. The final survival function is shown in Equation 2.h(t) is the hazard; h0(t) is the baseline hazard, −1.32 is the median ZHA of the population, and high income refers to the socioeconomic tertile with the highest income with low and middle income as the reference category, and η is the interindividual variability term.An exponential survival distribution best predicted malaria for the no chemoprevention and SP regimens Figure . A WeibuCompared to no chemoprevention, there was no difference in hazard of malaria in the SP arm, and these study arms were combined in the analysis. For the TS arm, although there was a 77% reduction in the hazard (HR [95% CI] 0.23 [0.18–0.31]) of malaria within a month of starting chemoprevention or developing an episode of malaria, compared to no chemoprevention or SP, this benefit decreased with time and the hazard of malaria for children who received TS was similar to those who received no chemoprevention or SP after 3.5 months of drug. Compared to no chemoprevention or SP, children who received DP had a lower risk of malaria throughout chemoprevention, but the benefit decreased over time, with a 35% lower hazard of malaria after 18 months of chemoprevention  decreased hazard of subsequent malaria (ΔOFV −5.6) compared to children whose most recent PQ concentration was less than or equal to 10.3 ng/ml. After accounting for PQ concentrations greater than 10.3 ng/ml, a ZHA less than −2 was associated with a twofold increased malaria hazard  compared to a ZHA of zero Table .All height, weight, and age measurements obtained after the start of chemoprevention were included in height and weight growth models. All significant covariate relationships were identified on the growth rate (KTR) of the Brody function. Cumulative prior malaria episodes , ZHA and WHZ at the start of chemoprevention, and HIV exposure were associated with height  studies of drug‐based malaria prevention interventions should prioritize enrolling malnourished children and they should also incorporate outcomes, such as measures of malnutrition to best evaluate the intervention's health impacts.2 similar to the 25% of children who were stunted at the start of chemoprevention in these studies. Chronic malnutrition, as measured by low ZHA, has been reported as a risk factor for malaria.Malnutrition is highly prevalent in Africa, including in this cohort of Ugandan children. The estimated national prevalence of stunting in Ugandan children less than 5 years of age is 28.5%,P. falciparum IgG antibody response, or lower anti‐malarial drug exposure due to insufficient weight‐based dosing or lower oral bioavailability.P. falciparum immunity, they will need even higher drug exposure for full malaria protection.P. falciparum, including children and primigravida pregnant women, demonstrated lower treatment responses for acute malaria or required higher drug concentrations for full malaria prevention.Multiple potential mechanisms for an association between malnutrition and malaria have been proposed, including malnutrition leading to reduced immunity to malaria by lowering anti‐Complementary to this finding, the Weibull distribution  best predicted malaria hazard in the TS and DP arms, whereas in the no chemoprevention and SP arms, the hazard was constant. These differences in malaria risk trajectory by the chemoprevention arm may have been due to lower adherence to the study drugs as the study progressed or insufficient weight‐based doses as the child aged resulting in suboptimal drug exposure. PQ concentrations decreased over time as efficacy decreased in the DP arm. Dose optimization to compensate for developmental changes in drug metabolism as well as adherence interventions could be needed to improve malaria chemoprevention.Other studies have suggested malnutrition could be a confounder for socioeconomic factors that could increase malaria risk .Although HIV‐exposed and ‐unexposed children received their care at the same study clinic and were enrolled over a similar period of time, HIV‐exposed children had a 21% lower risk of malaria compared to HIV‐unexposed children throughout the study period. A biological cause for decreased risk of malaria associated with HIV exposure is unlikely. Prior studies have found no difference in the burden of malaria between HIV‐exposed and HIV‐unexposed infants.We found each additional malaria episode was associated with a decreased subsequent growth rate for height by 0.4%. As malaria episodes increased, so did the risk of stunting by 3 years of age. Consistent with this finding, effective malaria prevention with LLIN has been associated with significant weight and length gains in children.This study has some limitations. First, we could not independently assess the role of breastfeeding on growth, due to confounding with HIV‐exposure. As measured, breast‐feeding did not improve growth predictions in our models, although this has been an important predictor in prior studies which assessed growth in sub‐Saharan African infants.In conclusion, this analysis supports a bidirectional relationship between malnutrition, as measured by ZHA, and an increased risk of recurrent malaria. We found that the relationship between increased risk of malaria with lower ZHA persisted despite the choice of chemoprevention regimen, even when the most efficacious malaria chemoprevention regimen (DP) was used. Our findings suggest that optimizing chemoprevention regimens for high‐risk groups like malnourished young children, will be essential to maximize efficacy of any chemoprevention option. Further research is needed to identify the optimal chemoprevention regimens for young children with malnutrition and to evaluate how treating chronic malnutrition during infancy could improve the burden of malaria and overall health outcomes in young children.A.M.A., E.W., E.H., G.D., and R.M.S. wrote the manuscript. A.M.A., E.W., E.H., G.D., and R.M.S. designed the research. A.M.A., E.W., and E.H. performed the research. A.M.A., E.W., R.M.S., and E.H. analyzed the data.This study was supported by the Bill and Melinda Gates Foundation OPP1191117, and the National Institutes of Health KL2 TR001870‐04.The authors declared no competing interests for this work.Appendix S1Click here for additional data file.Appendix S2Click here for additional data file.Appendix S3Click here for additional data file.Appendix S4Click here for additional data file.


Plasmodium falciparum  malaria on KSHV’s biphasic life cycle and lytic reactivation has not been determined. Here, we examined KSHV serological profiles and viral loads in 134 children with acute malaria and 221 healthy children from high malaria regions in Kisumu, as well as 77 healthy children from low malaria regions in Nandi. We assayed KSHV, Epstein–Barr virus (EBV), and P. falciparum malaria antibody responses in these three by multiplexed Luminex assay. We confirmed that KSHV seroprevalence was significantly associated with malaria endemicity  with 71–77% seropositivity in high-malaria (Kisumu) compared to 28% in low-malaria (Nandi) regions. Furthermore, KSHV serological profiles during acute malaria episodes were distinct from age-matched non-malaria-infected children from the same region. Paired IgG levels also varied after malaria treatment, with significantly higher anti-ORF59 at day 0 but elevated ORF38, ORF73, and K8.1 at day 3. Acute malaria episodes is characterized by perturbation of KSHV latency in seropositive children, providing further evidence that malaria endemicity contributes to the observed increase in endemic KS incidence in sub-Saharan Africa.The seroprevalence of Kaposi sarcoma-associated herpesvirus (KSHV) and the incidence of endemic Kaposi sarcoma (KS) overlap with regions of malaria endemicity in sub-Saharan Africa. Multiple studies have shown an increased risk of KSHV seroconversion in children from high malaria compared to low malaria regions; however, the impact of acute episodes of  Plasmodium falciparum  malaria infections with inducing KSHV lytic replication and by extension, the strong geographical clustering of malaria endemicity with KSHV seroprevalence, changes in KSHV serological profiles during acute malaria episodes have not yet been studied.Kaposi sarcoma-associated herpesvirus (KSHV) is an oncovirus that causes Kaposi sarcoma (KS) and other diseases, including primary effusion lymphoma and a form of multicentric Castleman disease (KSHV-MCD) . The serP. falciparum malaria and another herpesvirus, Epstein–Barr virus (EBV) in the etiology of endemic Burkitt lymphoma (eBL) have been extensively described [P. falciparum erythrocyte membrane protein 1 (PfEMP1) on parasitized red blood cells (pRBCs). This direct interaction stimulates B cell proliferation and EBV reactivation [PfEMP1 CIDR1ɑ domain has been implicated in KSHV lytic replication [By comparison, the interactions between escribed ,13,14. Sescribed ,16. Cogntivation ,18,19. Tlication ,8,20. ThIn this study, we evaluated KSHV serological patterns indicative of lytic replication in children during an episode of acute malaria and compared them to two separate groups of non-malaria-infected Kenyan children. We demonstrate that acute malaria subjects exhibit a distinct pattern of elevated antibodies against KSHV lytic antigens, indicating that malaria infection induces KSHV reactivation.P. falciparum malaria from Kisumu (A-KSM) were compared to 221  healthy children from Kisumu (H-KSM), and 77  healthy children from Nandi (H-NDI). As expected, KSHV seroprevalence correlated with levels of malaria transmission, with significantly higher seroprevalence in high-malaria Kisumu children compared to low-malaria Nandi children  as well as healthy subjects from a region with low incidence of malaria (Nandi). A total of 134 children  experiencing an episode of uncomplicated acute p = 0.01 and A-KSM OR = 1.95, 1.19–3.35, 95% CI, p = 0.01) and in children with high P. falciparum malaria anti-MSP1 IgG antibodies . We next compared the serological profiles of the three cohorts and found that IgG antibodies against malaria antigens, MSP1 and AMA1, were significantly elevated in KSHV seropositive compared to seronegative children a, a pattTo determine the effects of acute malaria infections on KSHV serological patterns, we first projected individual participants from the three study populations (independent of KSHV sero-status) into a 2-D principal component analysis (PCA) based on previously validated IgG antibody levels to seven KSHV antigens c. A combTo determine the specific KSHV antigens that are elevated during acute malaria, we compared antibody levels to these antigens in the three cohorts. Consistent with our PCA characterization, we identified distinct antibody signatures for the KSHV antigens ORF59, K8.1, ORF38, and ORF73 across the three study populations. IgG antibodies against ORF73 were significantly higher in healthy subjects compared to A-KSM-D0 subjects. K8.1 IgG antibodies were also slightly elevated in H-KSM while ORF59 IgG was slightly elevated in A-KSM-D0 subjects compared to H-KSM a. In conp < 0.0001), K8.1 (p < 0.0001), and ORF73 (p = 0.0002) antigens were significantly higher at day 3 compared to day 0  and post-treatment samples (A-KSM-D3). From the pairwise comparisons, IgG antibodies against ORF38  and low (Nandi) malaria transmission regions in Kenya. Our results show a distinct KSHV serological profile during an episode of malaria, reminiscent of KSHV DNA replication, followed by the release of structural virion proteins as well as the KSHV latency genes during virion-induced cell destruction. Our observation is consistent with a recently proposed model of molecular interaction between pRBCs and KSHV-infected memory B-cells [In this study, we report the first detailed comparison of a panel of KSHV antibodies in children with acute  B-cells . Using p B-cells .During acute malaria episodes, previous studies have shown that EBV-specific T cells fail to control EBV-infected B cells, leading to abnormal proliferation, initiation of lytic reactivation, and elevated viral loads ,23,24. IIn conclusion, this study provides evidence that acute malaria infections initiate a transition through KSHV lytic replication stages characterized by a serological pattern reflective of the expression of early intermediate, delayed early, and tegument lytic KSHV proteins. Thus, it also provides further evidence that malaria seems to be reactivating KSHV replication, possibly thereby contributing to the increased incidence of endemic KS in the region. Though the average decline in KS incidence is largely associated with a reduction in HIV/AIDS incidence and the rollout of antiretroviral therapy , the impP. falciparum malaria  were sampled from a larger multicenter study designed to evaluate the efficacy of artemether-lumefantrine (AL) in Ahero, Kisumu County, Kenya between September 2016 and July 2018. Participants were included in the study if they presented with at least 500 parasites per 200 white blood cells (WBC) by microscopy and admitted to Ahero sub-county hospital clinical trial site in Kisumu County for directly observed therapy consisting of 6 doses over 3 days, following the current Ministry of Health (Kenya) guidelines [Children presenting with acute uncomplicated idelines . Whole bidelines . For incUsing multiplexed bead-based assay, we measured the levels of IgG and IgM antibodies to 7 previously described KSHV antigens ,41 and aP. falciparum malaria quantification by real-time PCR (qPCR) amplification of the lactate dehydrogenase (pfldh) gene [ORF26 and BALF5 genes was used to quantify KSHV and EBV loads, respectively, relative to the human β-actin gene. Previously published ORF26 [BALF5 [To determine malaria parasite density and viral load, DNA was extracted from dry blood spots  using Qiagen QiaAMP DNA extraction kits  in accordance with the manufacturer’s instructions. DNA was suspended in the elution buffer, followed by dh) gene . Dropleted ORF26 , human β6 [BALF5  primers prcomp with a scaled option. All statistical analyses and data visualization were performed using R (version 3.6.3) and GraphPad Prism (v9.0.0). KSHV seropositivity cutoff was estimated using ROC analysis as previously described using MFI values from 11 African and 15 North American control samples . Wilcoxo