Comparative transcriptomic analysis reveals translationally relevant processes in mouse models of malaria.

Recent initiatives to improve translation of findings from animal models to human disease have focussed on reproducibility but quantifying the relevance of animal models remains a challenge. Here, we use comparative transcriptomics of blood to evaluate the systemic host response and its concordance between humans with different clinical manifestations of malaria and five commonly used mouse models. Plasmodium yoelii 17XL infection of mice most closely reproduces the profile of gene expression changes seen in the major human severe malaria syndromes, accompanied by high parasite biomass, severe anemia, hyperlactatemia, and cerebral microvascular pathology. However, there is also considerable discordance of changes in gene expression between the different host species and across all models, indicating that the relevance of biological mechanisms of interest in each model should be assessed before conducting experiments. These data will aid the selection of appropriate models for translational malaria research, and the approach is generalizable to other disease models.

Malaria Epidemiology Along the Indian Districts Bordering Bhutan and Implications for Malaria Elimination in the Region.

It is important for malaria-endemic countries to address malaria control across international borders, and in particular to prioritize appropriate rapid diagnosis, treatment and surveillance. Bhutan and India aim to achieve malaria elimination by 2023 and 2030 respectively. Malaria elimination along the Indo-Bhutan border is of common concern. We delineated malaria epidemiology along the border to provide a blueprint for focusing malaria control efforts in key foci within this region. Epidemiological data from 2015 to 2019 were analyzed, as the most drastic reductions in malaria burden across most parts of India were witnessed in this time frame. Several areas of concern include low surveillance in most border districts, favorable climatic conditions for perennial malaria transmission, and movement of potential parasite carriers because of the porous borders. India and Bhutan need to control the importation/exportation of malaria cases. We highlight the foci of concern for which implementing tailor-made malaria control strategies may benefit both countries.

PREVALENCE OF PARASITIC INFECTIONS IN CHILDREN OF BOKE, GUINEA.

Helminthic and intestinal protozoan infections and malaria infections are common in children less than 15 yr old in sub-Saharan Africa, but little is known about these infections in Guinea. The aim of this study was to determine the prevalence of parasitic infections in children aged less than 15 yr and the relationship of these infections with anemia. The cross-sectional study was done in Dabbis sub-prefecture in the Boke region of Guinea from 18 to 26 March 2017. A simple random sampling at the household level was performed, and 1 child under the age of 15 was included per eligible household. A total of 392 children were included in the analysis. Clinical and parasitological information were assessed, including anthropometric measures (weight and height), disease symptoms, hemoglobin level, and malaria parasitemia. Helminthic and protozoan intestinal infections were present in 59.7% of the children surveyed. Malaria infection prevalence was 45.5% when assessed by microscopy and 43.6% when assessed by a rapid diagnostic test. Plasmodium falciparum, accounting for 84.2% of malaria infections, was the main malaria species infection. Gastrointestinal parasites were present in 19.1% of children. The main gastrointestinal parasites present included Entamoeba coli (5.4%) and Giardia intestinalis (5.1%). There was no association between the presence of anemia and the parasitic status of the children. Parasitic screening and mass treatment in this age group, as well as household awareness raising, would reduce cases of parasitic infections in rural Guinea.

Burden of malaria in children under five and caregivers' health-seeking behaviour for malaria-related symptoms in artisanal mining communities in Ghana.

BACKGROUND: Artisanal mining creates enabling breeding ground for the vector of malaria parasites. There is paucity of data on the effects of artisanal mining on malaria. This study assessed burden of malaria and caregivers' health-seeking behaviour for children under five in artisanal mining communities in East Akim District in Ghana. METHODS: A cross-sectional study involving caregivers and their children under five was conducted in three artisanal mining communities in the East Akim District in Ghana. Caregivers were interviewed using a structured questionnaire. Finger prick blood samples were collected and analysed for haemoglobin concentration using a rapid diagnostic test, and thick and thin blood smears were analysed to confirm the presence of malaria parasites. RESULTS: Of the 372 children under 5 years included in the study, 197 (53.1%) were male, with a mean age (± SD) of 23.0 ± 12.7 months. The proportion of children with malaria (Plasmodium falciparum and P. malariae) was 98.1% and 1.9%, respectively, whilst the proportion with anaemia (Hb < 11.0 g/dl) was 39.5% (n = 147). Almost all caregivers were female (98.9%), and 28.6% (n = 106) did not have access to any malaria control information. Caregivers associated malaria infection with mosquito bites (68.3%, n = 254) and poor sanitation (21.2%, n = 79). Malaria in children under five was significantly associated with anaemia (OR 11.07, 95% CI 6.59-18.68, n = 111/160, 69.4%; P < 0.0001), residing close to stagnant water (≤ 25 m) from an artisanal mining site (AOR 2.91, 95% CI 1.47-5.76, P = 0.002) and caregiver age younger than 30 years (OR 0.44, 95% CI 0.208-0.917, n = 162, 43.55%, P = 0.001). CONCLUSIONS: There is a high burden of malaria and anaemia among children under five in artisanal mining communities of the East Akim District, and far higher than in non-artisanal mining sites. Interventions are needed to effectively regulate mining activities in these communities, and strengthen malaria control and health education campaigns to curtail the high malaria burden and improve health-seeking behaviour.

Quantification of the misidentification of Plasmodium knowlesi as Plasmodium malariae by microscopy: an analysis of 1569 P. knowlesi cases.

BACKGROUND: Plasmodium knowlesi is recognized as the fifth Plasmodium species causing malaria in humans. It is morphologically similar to the human malaria parasite Plasmodium malariae, so molecular detection should be used to clearly discriminate between these Plasmodium species. This study aimed to quantify the rate at which P. knowlesi is misidentified as P. malariae by microscopy in endemic and non-endemic areas. METHODS: The protocol of this systematic review was registered in the PROSPERO International Prospective Register of Systematic Reviews (ID = CRD42020204770). Studies reporting the misidentification of P. knowlesi as P. malariae by microscopy and confirmation of this by molecular methods in MEDLINE, Web of Science and Scopus were reviewed. The risk of bias in the included studies was assessed using the Quality Assessment of Diagnostic Accuracy Studies (QUADAS). The pooled prevalence and 95% confidence interval (CI) of the misidentification of P. knowlesi as P. malariae by microscopy were estimated using a random effects model. Subgroup analysis of the study sites was performed to demonstrate any differences in the misidentification rates in different areas. Heterogeneity across the included studies was assessed and quantified using Cochran's Q and I2 statistics, respectively. Publication bias in the included studies was assessed using the funnel plot, Egger's test and contour-enhanced funnel plot. RESULTS: Among 375 reviewed studies, 11 studies with a total of 1569 confirmed P. knowlesi cases in humans were included. Overall, the pooled prevalence of the misidentification of P. knowlesi as P. malariae by microscopy was estimated at 57% (95% CI 37-77%, I2: 99.3%). Subgroup analysis demonstrated the highest rate of misidentification in Sawarak, Malaysia (87%, 95% CI 83-90%, I2: 95%), followed by Sabah, Malaysia (85%, 95% CI 79-92%, I2: 85.1%), Indonesia (16%, 95% CI 6-38%), and then Thailand (4%, 95% CI 2-9%, I2: 95%). CONCLUSION: Although the World Health Organization (WHO) recommends that all P. malariae-positive diagnoses made by microscopy in P. knowlesi endemic areas be reported as P. malariae/P. knowlesi malaria, the possibility of microscopists misidentifying P. knowlesi as P. malariae is a diagnostic challenge. The use of molecular techniques in cases with malariae-like Plasmodium with high parasite density as determined by microscopy could help identify human P. knowlesi cases in non-endemic countries.

First evidence of local circulation of Plasmodium ovale curtisi and reliability of a malaria rapid diagnostic test among symptomatic outpatients in Douala, Cameroon.

The performances of a commonly used Plasmodium falciparum-detecting rapid diagnostic test (RDT) were determined in symptomatic individuals living in Cameroon. Discrepancies between RDT and light microscopy (LM) results were further investigated, with a focus on non-falciparum malaria (NFM) which are still largely understudied in sub-Saharan Africa (sSA) countries. In the present study, a total of 355 individuals aged 1-65 years were enrolled in the study. Their signs/symptoms and sociodemographic characteristics were documented. The RDT reliability was evaluated using LM as gold standard method. Polymerase chain reaction (PCR) of Plasmodium 18S gene was performed for samples with discordant results between LM and RDT (i.e., RDT-/LM+, and RDT+/LM-). The PCR amplicons of NFM species were sequenced and BLASTed. The prevalence of malaria infection by LM was 95.7% (95% CI: 93.1-97.4%). The sensitivity and specificity of RDT for P. falciparum detection was 94.0% and 66.7%, respectively. By PCR assay, P. ovale curtisi (PoC) was found in 5 of the 30 discordant samples, and on sequence analysis these isolates were found to be phylogenetically closer to sequences reported from China-Myanmar border and Malaysia. This is the first report on molecular characterization of P. ovale subspecies in Cameroon. The study also outlines the good diagnostic performances of the RDT for detection of P. falciparum. Though, the presence of PoC indicated the importance of having RDTs targeting the NFM species in malaria diagnosis and treatment, which is presently limited in the country.

Estimating the Fraction of Severe Malaria among Malaria-Positive Children: Analysis of Household Surveys in 19 Malaria-Endemic Countries in Africa.

To date, the only robust estimates of severe malaria cases include children who present to the formal healthcare system. It is a challenge to use these data because of varying age ranges of reporting, different diagnosis techniques, surveillance methods, and healthcare utilization. This analysis examined data from 37 Demographic and Health Surveys and Malaria Indicator Surveys across 19 countries in sub-Saharan Africa collected between 2011 and 2018. The outcome of interest is a proxy indicator for severe malaria, defined as a proportion of children aged 6-59 months with at least one self-reported symptom of severe illness including loss of consciousness, rapid breathing, seizures, or severe anemia (hemoglobin < 5 g/dL) among those who were positive for malaria. The study includes a weighted descriptive, country-level analysis and a multilevel mixed-effects logistic regression model to assess the determinants of severe malaria. Among children positive for malaria across all surveys, 4.5% (95% CI: 4.1-4.8) had at least one sign or symptom of severe malaria, which was significantly associated with age, residence, wealth, and year of survey fieldwork at a P-value less than 0.05. This analysis presents a novel and an alternative approach of estimating the fraction of severe malaria cases among malaria-positive children younger than 5 years in malaria-endemic countries. Estimating severe malaria cases through population-based surveys allows countries to estimate severe malaria across time and to compare with other countries. Having a population-level estimate of severe malaria cases helps further our understanding of the burden and epidemiology of severe malaria.

THE TERMINAL APHONIA OF ALEXANDER THE GREAT, A SILENT HERALD OF AN ORDINARY DEATH

Even though the absence of the body prevents sure conclusions, the death of Alexander the Great remains a hot topic of retrospective diagnosis. Due to the serious mishandling of ancient sources, the scientific literature had Alexander dying of every possible natural cause. In previous works, the hypothesis that typhoid fever killed Alexander was proposed, based on the presence of the remittent fever typical of this disease in the narrations of Plutarch and Arrian. Here we provide additional evidence for the presence of stupor, the second distinctive symptom of typhoid fever. In fact, based on the authority of Caelius Aurelianus and Galen, we demonstrate that the word ἄφωνος, used to describe the last moments of Alexander, is a technical word of the lexicon of the pathology of Hippocrates. Used by him, the word defines a group of diseases sharing a serious depression of consciousness and motility. The association of stupor with the remittent fever strengthens the typhoid fever hypothesis.

Predictors of malaria rapid diagnostic test positivity in a high burden area of Paletwa Township, Chin State in Western Myanmar.

BACKGROUND: Despite major reductions in malaria burden across Myanmar, clusters of the disease continue to persist in specific subregions. This study aimed to assess the predictors of test positivity among people living in Paletwa Township of Chin State, an area of persistently high malaria burden. METHODS: Four villages with the highest malaria incidence from Paletwa Township were purposively selected. The characteristics of 1045 subjects seeking malaria diagnosis from the four assigned village health volunteers from January to December, 2018 were retrospectively analyzed. Their household conditions and surroundings were also recorded using a checklist. Descriptive statistics and logistic regression models were applied to investigate potential associations between individual and household characteristics and malaria diagnosis. RESULTS: In 2017, the Paletwa township presented 20.9% positivity and an annual parasite index of 46.9 cases per 1000 people. Plasmodium falciparum was the predominant species and accounted for more than 80.0% of all infections. Among 1045 people presenting at a clinic with malaria symptoms, 31.1% were diagnosed with malaria. Predictors for test positivity included living in a hut [adjusted odds ratios (a OR): 2.3, 95% confidence intervals (CI): 1.2-4.6], owning farm animals (aOR: 1.7, 95% CI: 1.1-3.6), using non-septic type of toilets (aOR: 1.9, 95% CI: 1.1-8.4), presenting with fever (aOR: 1.9, 95% CI: 1.1-3.0), having a malaria episode within the last year (aOR: 2.9, 95% CI: 1.4-5.8), traveling outside the village in the previous 14 days (aOR: 4.5, 95% CI: 1.5-13.4), and not using bed nets (a OR: 3.4, 95% CI: 2.3-5.1). There were no statistically significant differences by age or gender in this present analysis. CONCLUSIONS: The results from this study, including a high proportion of P. falciparum infections, little difference in age, sex, or occupation, suggest that malaria is a major burden for these study villages. Targeted health education campaigns should be introduced to strengthen synchronous diagnosis-seeking behaviors, tighten treatment adherence, receiving a diagnosis after traveling to endemic regions, and using bed nets properly. We suggest increased surveillance, early diagnosis, and treatment efforts to control the disease and then to consider the local elimination.

Significant symptoms and nonsymptom-related factors for malaria diagnosis in endemic regions of Indonesia.

OBJECTIVES: This study aims to identify significant symptoms and nonsymptom-related factors for malaria diagnosis in endemic regions of Indonesia. METHODS: Medical records are collected from patients suffering from malaria and other febrile diseases from public hospitals in endemic regions of Indonesia. Interviews with eight Indonesian medical doctors are conducted. Feature selection and machine learning techniques are used to develop malaria classifiers for identifying significant symptoms and nonsymptom-related factors. RESULTS: Seven significant symptoms (duration of fever, headache, nausea and vomiting, heartburn, severe symptom, dizziness, and joint pain) and patients' history of malaria as a nonsymptom-related factor contribute most to malaria diagnosis. As a symptom, fever duration is more significant than temperature or fever for distinguishing malaria from other febrile diseases. Shivering, fever, and sweating (known to indicate malaria presence in Indonesia) are shown to be less significant than other symptoms in endemic regions. CONCLUSIONS: Three most suitable malaria classifiers have been developed to identify the significant features that can be used to predict malaria as distinct from other febrile diseases. With extensive experiments on the classifiers, the significant features identified can help medical doctors in the clinical diagnosis of malaria and raise public awareness of significant malaria symptoms at early stages.

Epidemiological trends of malaria in the Western regions of Saudi Arabia: a cross sectional study.

INTRODUCTION: Saudi Arabia has successfully reduced malaria cases to be constrained largely in the western regions. This study aimed to determine the epidemiological trends of malaria infection in five western regions of Saudi Arabia. METHODOLOGY: A retrospective analysis was conducted to investigate the epidemiological trends of malaria infection in the western regions, based on the published registry of the Saudi Ministry of Health, during the period from 2014 to 2017 using the appropriate statistical tools. RESULTS: A total of 8925 confirmed cases of malaria were reported in the western regions during the period from 2014 to 2017 with the mean of 2231 malaria cases per year. The minimum (n = 1097) and maximum (n = 4075) number of cases were reported in 2014 and 2016 respectively. The highest (n = 5919, 66.3%) number of cases were reported from Jazan region, while lowest (n = 86, 1.0%) number of cases were reported from Al-Bahah region. Plasmodium falciparum was the most frequently reported species with 7485 (83.9%) cases, while Plasmodium vivax accounted 1386 (15.5%) cases. Plasmodium malariae and mixed infections were insignificant and accounted 0.5% (n = 48) and 0.1% (n = 6) cases respectively. In relation to malaria infection and age group, malaria was predominant in > 15 age group. The highest number of malaria cases in almost all years was observed from January until March and the lowest number was reported from May until July. CONCLUSIONS: Plasmodium falciparum was the most dominant species in this survey and Jazan was the most affected region.

Analysis of initial laboratory diagnosis of malaria and its accuracy compared with re-testing from 2013 to 2018 in Yunnan Province, China.

BACKGROUND: According to China's Malaria Eradication Action Plan, malaria cases diagnosed and reported by health authorities at the county level must be further re-confirmed by provincial laboratories. The Yunnan Province Malaria Diagnostic Reference Laboratory (YPMDRL) began the synchronous implementation of microscopic examinations and nested polymerase chain reaction (nested-PCR) testing to re-test the malaria cases initially diagnosed by county-level laboratories and to evaluate the consistency of Plasmodium species identified between by YPMDRL and by the county-level laboratories from 2013 to 2018 in Yunnan Province. METHODS: Data on malaria initial diagnosis completed by county-level laboratories in Yunnan Province were collected weekly from the "China Disease Prevention and Control Information System" from 2013 to 2018. The YPMDRL performed Plasmodium microscopic examination and 18S rRNA gene nested-PCR testing on every malaria case managed by the China Disease Prevention and Control Information System. The re-testing detection results were fed back to the initial diagnosis and reporting unit for revision of malaria case types. RESULTS: A total of 2,869 malaria cases were diagnosed and reported by county-level laboratories in Yunnan Province from 2013 to 2018. The re-testing rate was 95.6% (2,742/2,869), and the re-testing rate increased from 2013 to 2018. Among the re-tested 2,742 cases, 96.7% (2651/2742), 2.2% (59/2742), and 1.1% (32/2742) were doubly examined by microscopy and by nested-PCR, only by microscopy, and only by nested-PCR, respectively. The total Plasmodium species accuracy rate at county-level laboratories was 92.6% (2,543/2,742) reference to the diagnosis by YPMDRL. Among the inconsistent 199 cases, they were identified as including 103 negative cases, 45 falciparum malaria cases, 30 vivax malaria cases, 11 ovale malaria cases, and 10 malariae malaria cases by YPMDRL. From 2013 to 2018, the revised and registered malaria cases by the China Disease Prevention and Control Information System in Yunnan Province was 2,747 cases, including 2,305 vivax malaria cases, 421 falciparum malaria cases, 11 ovale malaria cases, and 10 malariae malaria cases. CONCLUSIONS: The double re-testing strategy by microscopy and by gene testing increases the accuracy of diagnoses malaria in Yunnan Province, and gene testing can reliably differentiate Plasmodium species. The re-testing results provided by YPMDRL are the authoritative basis for revising malaria kind in Yunnan Province.

Identification of French Guiana anopheline mosquitoes by MALDI-TOF MS profiling using protein signatures from two body parts.

In French Guiana, the malaria, a parasitic infection transmitted by Anopheline mosquitoes, remains a disease of public health importance. To prevent malaria transmission, the main effective way remains Anopheles control. For an effective control, accurate Anopheles species identification is indispensable to distinguish malaria vectors from non-vectors. Although, morphological and molecular methods are largely used, an innovative tool, based on protein pattern comparisons, the Matrix Assisted Laser Desorption / Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) profiling, emerged this last decade for arthropod identification. However, the limited mosquito fauna diversity of reference MS spectra remains one of the main drawback for its large usage. The aim of the present study was then to create and to share reference MS spectra for the identification of French Guiana Anopheline species. A total of eight distinct Anopheles species, among which four are malaria vectors, were collected in 6 areas. To improve Anopheles identification, two body parts, legs and thoraxes, were independently submitted to MS for the creation of respective reference MS spectra database (DB). This study underlined that double checking by MS enhanced the Anopheles identification confidence and rate of reliable classification. The sharing of this reference MS spectra DB should make easier Anopheles species monitoring in endemic malaria area to help malaria vector control or elimination programs.

A two-colour multiplexed lateral flow immunoassay system to differentially detect human malaria species on a single test line.

BACKGROUND: Malaria continues to impose a tremendous burden in terms of global morbidity and mortality, yet even today, a large number of diagnoses are presumptive resulting in lack of or inappropriate treatment. METHODS: In this work, a two-colour lateral flow immunoassay (LFA) system was developed to identify infections by Plasmodium spp. and differentiate Plasmodium falciparum infection from the other three human malaria species (Plasmodium vivax, Plasmodium ovale, Plasmodium malariae). To achieve this goal, red and blue colours were encoded to two markers on a single test line of strips, for simultaneous detection of PfHRP2 (red), a marker specific for P. falciparum infection, and pLDH (blue), a pan-specific marker for infections by all species of Plasmodium. The assay performance was first optimized and evaluated with recombinant malarial proteins spiked in washing buffer at various concentrations from 0 to 1000 ng mL-1. The colour profiles developed on the single test line were discriminated and quantified: colour types corresponded to malaria protein species; colour intensities represented protein concentration levels. RESULTS: The limit of detection (the lowest concentrations of malaria antigens that can be distinguished from blank samples) and the limit of colour discrimination (the limit to differentiate pLDH from PfHRP2) were defined for the two-colour assay from the spiked buffer test, and the two limits were 31.2 ng mL-1 and 7.8 ng mL-1, respectively. To further validate the efficacy of the assay, 25 human whole blood frozen samples were tested and successfully validated against ELISA and microscopy results: 15 samples showed malaria negative; 5 samples showed P. falciparum positive; 5 samples showed P. falciparum negative, but contained other malaria species. CONCLUSIONS: The assay provides a simple method to quickly identify and differentiate infection by different malarial parasites at the point-of-need and overcome the physical limitations of traditional LFAs, improving the multiplexing potential for simultaneous detection of various biomarkers.

[Management of malaria in Sweden]. / Handläggning av malaria ­ ur ett svenskt perspektiv.

Malaria is a potentially severe infection and time to treatment can be decisive for the outcome. Febrile patients returning from travel in endemic areas should therefore be promptly investigated for malaria. This review focuses on the acute management of malaria in Sweden. The disease is diagnosed in travellers, migrants and temporary visitors from malaria-endemic countries. Malaria is a relatively rare infection in Sweden, with approximately 150 imported cases per year in a population of 10 million. Health care delay is a risk of more severe disease. Children, pregnant women, elderly, and individuals from endemic areas who lived in Sweden for a long time as well as those with comorbidities are at increased risk of severe malaria. Microscopy is used for diagnosis and determination parasite density; rapid diagnostic tests are supportive diagnostic tools. First-line treatment for severe malaria is intravenous artesunate and for uncomplicated P. falciparum malaria artemether-lumefantrine (AL) or chloroquine in cases with non-P. falciparum infections from areas without known resistance. Treatment failures have been observed in non-immune travelers treated with AL, and patients should be recommended to seek care in the event of new fever. Being a relative rare disease in Sweden, management of malaria is performed at specialized centers with infectious disease specialists.

A comparison of two PCR protocols for the differentiation of Plasmodium ovale species and implications for clinical management in travellers returning to Germany: a 10-year cross-sectional study.

BACKGROUND: To assess the occurrence of Plasmodium ovale wallikeri and Plasmodium ovale curtisi species in travellers returning to Germany, two real-time PCR protocols for the detection and differentiation of the two P. ovale species were compared. Results of parasite differentiation were correlated with patient data. METHODS: Residual nucleic acid extractions from EDTA blood samples of patients with P. ovale spp. malaria, collected between 2010 and 2019 at the National Reference Centre for Tropical Pathogens in Germany, were subjected to further parasite discrimination in a retrospective assessment. All samples had been analysed by microscopy and by P. ovale spp.-specific real-time PCR without discrimination on species level. Two different real-time PCR protocols for species discrimination of P. o. curtisi and P. o. wallikeri were carried out. Results were correlated with patient data on gender, age, travel destination, thrombocyte count, and duration of parasite latency. RESULTS: Samples from 77 P. ovale spp. malaria patients were assessed, with a male:female ratio of about 2:1 and a median age of 30 years. Parasitaemia was low, ranging from few visible parasites up to 1% infected erythrocytes. Discriminative real-time PCRs revealed 41 cases of P. o. curtisi and 36 cases of P. o. wallikeri infections. Concordance of results by the two PCR approaches was 100%. Assessment of travel destinations confirmed co-existence of P. o. curtisi and P. o. wallikeri over a wide range of countries in sub-Saharan Africa. Latency periods for the two P. ovale species were similar, with median values of 56.0 days for P. o. curtisi and 58.0 days for P. o. wallikeri; likewise, there was no statistically significant difference in thrombocyte count with median values of 138.5/µL for patients with P. o. curtisi and 152.0/µL for P. o. wallikeri-infected patients. CONCLUSIONS: Two different real-time PCR protocols were found to be suitable for the discrimination of P. o. curtisi and P. o. wallikeri with only minor differences in sensitivity. Due to the overall low parasitaemia and the lack of differences in severity-related aspects like parasite latency periods or thrombocyte counts, this study supports the use of P. ovale spp. PCR without discrimination on species level to confirm the diagnosis and to inform clinical management of malaria in these patients.

Multiplex serology demonstrate cumulative prevalence and spatial distribution of malaria in Ethiopia.

BACKGROUND: Measures of malaria burden using microscopy and rapid diagnostic tests (RDTs) in cross-sectional household surveys may incompletely describe the burden of malaria in low-transmission settings. This study describes the pattern of malaria transmission in Ethiopia using serological antibody estimates derived from a nationwide household survey completed in 2015. METHODS: Dried blood spot (DBS) samples were collected during the Ethiopian Malaria Indicator Survey in 2015 from malarious areas across Ethiopia. Samples were analysed using bead-based multiplex assays for IgG antibodies for six Plasmodium antigens: four human malaria species-specific merozoite surface protein-1 19kD antigens (MSP-1) and Apical Membrane Antigen-1 (AMA-1) for Plasmodium falciparum and Plasmodium vivax. Seroprevalence was estimated by age, elevation and region. The seroconversion rate was estimated using a reversible catalytic model fitted with maximum likelihood methods. RESULTS: Of the 10,278 DBS samples available, 93.6% (9622/10,278) had valid serological results. The mean age of participants was 15.8 years and 53.3% were female. National seroprevalence for antibodies to P. falciparum was 32.1% (95% confidence interval (CI) 29.8-34.4) and 25.0% (95% CI 22.7-27.3) to P. vivax. Estimated seroprevalences for Plasmodium malariae and Plasmodium ovale were 8.6% (95% CI 7.6-9.7) and 3.1% (95% CI 2.5-3.8), respectively. For P. falciparum seroprevalence estimates were significantly higher at lower elevations (< 2000 m) compared to higher (2000-2500 m) (aOR 4.4; p < 0.01). Among regions, P. falciparum seroprevalence ranged from 11.0% (95% CI 8.8-13.7) in Somali to 65.0% (95% CI 58.0-71.4) in Gambela Region and for P. vivax from 4.0% (95% CI 2.6-6.2) in Somali to 36.7% (95% CI 30.0-44.1) in Amhara Region. Models fitted to measure seroconversion rates showed variation nationally and by elevation, region, antigen type, and within species. CONCLUSION: Using multiplex serology assays, this study explored the cumulative malaria burden and regional dynamics of the four human malarias in Ethiopia. High malaria burden was observed in the northwest compared to the east. High transmission in the Gambela and Benishangul-Gumuz Regions and the neglected presence of P. malariae and P. ovale may require programmatic attention. The use of a multiplex assay for antibody detection in low transmission settings has the potential to act as a more sensitive biomarker.

Collaborative intelligence and gamification for on-line malaria species differentiation.

BACKGROUND: Current World Health Organization recommendations for the management of malaria include the need for a parasitological confirmation prior to triggering appropriate treatment. The use of rapid diagnostic tests (RDTs) for malaria has contributed to a better infection recognition and a more targeted treatment. Nevertheless, low-density infections and parasites that fail to produce HRP2 can cause false-negative RDT results. Microscopy has traditionally been the methodology most commonly used to quantify malaria and characterize the infecting species, but the wider use of this technique remains challenging, as it requires trained personnel and processing capacity. OBJECTIVE: In this study, the feasibility of an on-line system for remote malaria species identification and differentiation has been investigated by crowdsourcing the analysis of digitalized infected thin blood smears by non-expert observers using a mobile app. METHODS: An on-line videogame in which players learned how to differentiate the young trophozoite stage of the five Plasmodium species has been designed. Images were digitalized with a smartphone camera adapted to the ocular of a conventional light microscope. Images from infected red blood cells were cropped and puzzled into an on-line game. During the game, players had to decide the malaria species (Plasmodium falciparum, Plasmodium malariae, Plasmodium vivax, Plasmodium ovale, Plasmodium knowlesi) of the infected cells that were shown in the screen. After 2 months, each player's decisions were analysed individually and collectively. RESULTS: On-line volunteers playing the game made more than 500,000 assessments for species differentiation. Statistically, when the choice of several players was combined (n > 25), they were able to significantly discriminate Plasmodium species, reaching a level of accuracy of 99% for all species combinations, except for P. knowlesi (80%). Non-expert decisions on which Plasmodium species was shown in the screen were made in less than 3 s. CONCLUSION: These findings show that it is possible to train malaria-naïve non-experts to identify and differentiate malaria species in digitalized thin blood samples. Although the accuracy of a single player is not perfect, the combination of the responses of multiple casual gamers can achieve an accuracy that is within the range of the diagnostic accuracy made by a trained microscopist.

Prospective comparative multi-centre study on imported Plasmodium ovale wallikeri and Plasmodium ovale curtisi infections.

BACKGROUND: Few previous retrospective studies suggest that Plasmodium ovale wallikeri seems to have a longer latency period and produces deeper thrombocytopaenia than Plasmodium ovale curtisi. Prospective studies were warranted to better assess interspecies differences. METHODS: Patients with imported P. ovale spp. infection diagnosed by thick or thin film, rapid diagnostic test (RDT) or polymerase chain reaction (PCR) were recruited between March 2014 and May 2017. All were confirmed by DNA isolation and classified as P. o. curtisi or P. o. wallikeri using partial sequencing of the ssrRNA gene. Epidemiological, analytical and clinical differences were analysed by statistical methods. RESULTS: A total of 79 samples (35 P. o. curtisi and 44 P. o. wallikeri) were correctly genotyped. Males predominate in wallikeri group (72.7%), whereas were 48.6% in curtisi group. Conversely, 74.3% of curtisi group were from patients of African ethnicity, whilst 52.3% of Caucasians were infected by P. o. wallikeri. After performing a multivariate analysis, more thrombocytopaenic patients (p = 0.022), a lower number of platelets (p = 0.015), a higher INR value (p = 0.041), and shorter latency in Caucasians (p = 0.034) were significantly seen in P. o. wallikeri. RDT sensitivity was 26.1% in P. o. curtisi and 42.4% in P. o. wallikeri. Nearly 20% of both species were diagnosed only by PCR. Total bilirubin over 3 mg/dL was found in three wallikeri cases. Two patients with curtisi infection had haemoglobin under 7 g/dL, one of them also with icterus. A wallikeri patient suffered from haemophagocytosis. Chemoprophylaxis failed in 14.8% and 35% of curtisi and wallikeri patients, respectively. All treated patients with various anti-malarials which included artesunate recovered. Diabetes mellitus was described in 5 patients (6.32%), 4 patients of wallikeri group and 1 curtisi. CONCLUSIONS: Imported P. o. wallikeri infection may be more frequent in males and Caucasians. Malaria caused by P. o. wallikeri produces more thrombocytopaenia, a higher INR and shorter latency in Caucasians and suggests a more pathogenic species. Severe cases can be seen in both species. Chemoprophylaxis seems less effective in P. ovale spp. infection than in P. falciparum, but any anti-malarial drug is effective as initial treatment. Diabetes mellitus could be a risk factor for P. ovale spp. infection.

Epidemiology of Malaria in the city of Kaedi (Mauritania, 2014).

BACKGROUND: In the face of ongoing and projected environmental and climate change, the epidemiology of malaria in the city of Kaédi (Mauritania), bordering the Senegal River Valley, requires special attention. OBJECTIVE: To describe the epidemiological situation of malaria in the city of Kaédi, Mauritania, during the wet season of 2014. METHODS: We conducted a descriptive cross-sectional survey in the city of Kaédi in september 2014 (wet season), to assess the prevalence of malaria parasites and vectors. For the choice of households, a cluster sampling was carried out and the city was subdivided into 10 sub-spatial units using a map of the city and the contribution of local populations. All household members were subjected to microscopic examination. In addition, larval surveys, morning wildlife sprays and night trap breaks were conducted. RESULTS: Of the 4671 thick drops made, three were positive, ie an average plasmid index of 0.06%. Prevalence was 0.04% (2/4671) and 0.02% (1/4671) for Plasmodium malariae and Plasmodium falciparum, respectively. In addition, the larval fauna consisted essentially of Culex larvae (100%). Two (2) female Anopheles mosquitoes were collected during the study. CONCLUSION: Even if transmission is low, in a context of absence of rainfall, the health authorities must foresee a strategy of malaria pre-elimination in riparian wilayas of the Senegal River.