Anti-malarial plants in Ethiopia and their activities on drug resistant malaria

In Ethiopia, the impacts of malaria continue to cause a many number of morbidity and mortality that accounts to most outpatient observations. Ethiopia recently designed to attain nationwide malaria control by 2030 by beginning sub-national elimination in districts with low malaria transmission. However, the rises of drug-resistant parasites, especially Plasmodium falciparum hinder the malaria containment strategies. Plasmodium falciparum and Plasmodium vivax, dispersed all over the Ethiopia and accounting for 60% and 40% of malaria cases respectively. The aim of this report was to overview the phytochemical constituents, diversity and effect of some compound extracts on drug resistant plasmodium species. Many plant species, a total 200 identified by 82 studies, are used in traditional malaria treatments throughout the country. Allium sativum, Croton macrostachyus and Carica papaya were the more frequently used medicinal plants species. There are so many phytochemicals constituents found in medicinal plants used to treat malaria. Alkaloids, Flavonoids, Phenolics, Terpenoid and Glycosides are the most reported for their effective activity on drug resistant malaria.

Entomological survey in two communes with residual malaria transmission in Gia Lai Province in the central highlands of Vietnam

Background In 2018, the National Malaria Control Programme in Vietnam switched from prioritizing malaria control to elimination. However, with the ongoing elimination programme, there are still areas where residual malaria transmission persists, including the central highlands. This entomological survey was conducted to evaluate Anopheles diversity and host-seeking activity of Anopheles vectors in two communes with very low malaria transmission in Gia Lai Province. Methods Anopheles species were collected in Ia DReh commune and Ia KDam commune, Gia Lai Province in the central highlands of Vietnam. Collections were conducted using human-baited double net trap, light trap and manual aspiration collections around cattle sheds, in the dry and rainy season. Mosquito specimens were identified morphologically, and members of species complexes were distinguished molecularly. Mosquito night-feeding patterns were investigated during the dry and rainy seasons. Results Overall, 18,835 specimens including 19 taxa were collected in Ia KDam and Ia DReh communes. These included the primary malaria vectors, Anopheles dirus and Anopheles minimus, and other secondary vector species. Anopheles dirus was observed to be an anthropophilic species, whereas An. minimus and a number of secondary vectors were observed to be zoophilic. Anopheles vagus was the dominant species, followed by Anopheles sinensis and Anopheles peditaeniatus. The majority of specimens were collected in the rainy season due to the relatively large number of An. vagus, while An. peditaeniatus, An. dirus, Anopheles kochi, Anopheles monstrosus and Anopheles tessellatus were collected in greater numbers during the dry season. The peak of host-seeking activity for An. dirus, An. sinensis, and An. vagus was between 18.00 and 19.00 h. Conclusion This study provided information on the diversity, seasonal prevalence and behaviour of Anopheles at the study sites. Identifying the diverse mosquito fauna in the central highlands of Vietnam allows species-specific control measures to be implemented by the National Programme to reduce malaria in areas of very low malaria transmission. The peak Anopheles host-seeking activity observed in this study was between 18.00 and 23.00 h, which highlights the need to better characterize Anopheles behaviour in this region of Vietnam to inform on vector control strategies.

GENETIC ARCHITECTURE AND DYNAMICS OF PFKELCH13S PROPELLER DOMAIN IN SENEGALESE PLASMODIUM FALCIPARUMCLINICAL ISOLATES

Plasmodium resistance to Artemisinin Combination-based Therapies (ACT) in Southeast Asia is a major public health concern that is sporadically appearing in Africa. Senegal has shifted from malaria control to elimination plans. Given notable progresses obtained through robust strategic plans,it is still crucial to assess genetic variability of the Plasmodium falciparumartemisinin resistance gene marker Kelch13 (PfKelch13)in circulating field isolates.We herereportan analysis of PfKelch13-propeller polymorphism in clinical isolates collected nine years after ACT introduction in five Senegalese regions with different malaria transmission settings. Sequencing of PfKelch13-propeller domainfrom 280 clinicalisolates reveals that 16% (45/280) of the parasite population harboredvariants. Dynamics of PfKelch13 variants reveals emerging, persistent but also disappearing mutations over time. In addition to the malaria epidemiology, our survey also shows the dynamics of PfKelch13 variants in different malaria transmission settings in Senegal. Despite the absence of PfKelch13associatedartemisinin resistance mutations, a shift from 86% to 68% of PfKelch13WTwas observed when comparing parasites collected prior vs. post ACT intensive usage in Dakar a low malaria transmission area. All together,our data confirms the need to closely monitor PfKelch13 polymorphism to anticipate and or preventemergence ofP. falciparum resistancein Senegal.

Quantification of the Dynamics of Antibody Response to Malaria to Inform Sero-Surveillance in Pregnant Women

Background: Malaria remains a major public health threat and in low malaria transmission areas new tools are needed to detect infections for prompt treatment and to progress elimination efforts. Pregnant women are particularly vulnerable to malaria infections and access routine antenatal care, presenting a unique sentinel population to apply novel sero-surveillance tools to measure malaria transmission. The aim of this study was to quantify the dynamic antibody responses to multiple antigens during pregnancy to identify a single or multiple antibody response of exposure to malaria in pregnancy. Methods: Antibody-mediated immunity responses to six parasite antigens (five commonly studied merozoite antigens and the variant surface antigen 2-chondroitin sulphate A (VAR2CSA), a pregnancy-specific erythrocytic antigen) were measured over the gestation period until delivery (median of 7 measurements/woman) in 250 pregnant women who attended antenatal clinics located at the Thai-Myanmar border. A multivariate mixture linear mixed model was used to cluster the pregnant women into groups that have similar longitudinal antibody responses to all six antigens over the gestational period using a Bayesian approach. The variable-specific entropy was calculated to identify the antibody responses that have the highest influence on the classification of the women into clusters, and subsequent agreement with grouping of women based on exposure to malaria during pregnancy. Results: Of the 250 pregnant women, 135 had a Plasmodium infection detected by light microscopy during pregnancy, defined as cases. The antibody responses to all six antigens accurately identified the women who did not have a malaria infection detected during pregnancy (93%, 107/115 controls). Antibody responses to P. falciparum merozoite surface protein 3 (PfMSP3) and P. vivax apical membrane antigen 1 (PvAMA1) were the least dynamic. Antibody responses to the antigens P. falciparum apical membrane antigen 1 (PfAMA1) and PfVAR2CSA were able to identify the majority of the cases more accurately (63%, 85/135). Conclusion: These findings suggest that the combination of antibodies, PfAMA1 and PfVAR2CSA, may be useful for sero-surveillance of malaria infections in pregnant women, particularly in low malaria transmission settings, leading to the early detection and treatment of malaria infections in pregnant women.

Asymptomatic malaria and anaemia among pregnant women during high and low malaria transmission seasons in Burkina Faso: household-based cross-sectional surveys in Burkina Faso, 2013 and 2017

Background Malaria in endemic countries is often asymptomatic during pregnancy, but it has substantial consequences for both the mother and her unborn baby. During pregnancy, anaemia is an important consequence of malaria infection. In Burkina Faso, the intensity of malaria varies according to the season, albeit the prevalence of malaria and anaemia as well as their risk factors, during high and low malaria transmission seasons is underexplored at the household level. Methods Data of 1751 pregnant women from October 2013 to March 2014 and 1931 pregnant women from April 2017 to June 2017 were drawn from two cross-sectional household surveys conducted in 24 health districts of Burkina Faso. Pregnant women were tested for malaria in their household after consenting. Asymptomatic carriage was defined as a positive result from malaria rapid diagnostic tests in the absence of clinical symptoms of malaria. Anaemia was defined as haemoglobin level less than 11 g/dL in the first and third trimester and less than 10.5 g/dL in the second trimester of pregnancy. Results Prevalence of asymptomatic malaria in pregnancy was estimated at 23.9% (95% CI 20.2–28.0) during the high transmission season (October–November) in 2013. During the low transmission season, it was 12.7% (95% CI 10.9–14.7) between December and March in 2013–2014 and halved (6.4%; 95% CI 5.3–7.6) between April and June 2017. Anaemia prevalence was estimated at 59.4% (95% CI 54.8–63.8) during the high transmission season in 2013. During the low transmission season, it was 50.6% (95% CI 47.7–53.4) between December and March 2013–2014 and 65.0% (95% CI 62.8–67.2) between April and June, 2017. Conclusion This study revealed that the prevalence of malaria asymptomatic carriage and anaemia among pregnant women at the community level remain high throughout the year. Thus, more efforts are needed to increase prevention measures such as IPTp–SP coverage in order to reduce anaemia and contribute to preventing low birth weight and poor pregnancy outcomes.

Ten years of monitoring malaria trend and factors associated with malaria test positivity rates in Lower Moshi

Background High altitude settings in Eastern Africa have been reported to experience increased malaria burden due to vector habitat expansion. This study explored possible associations between malaria test positivity rates and its predictors including malaria control measures and meteorological factors at a high-altitude, low malaria transmission setting, south of Mount Kilimanjaro. Methods Malaria cases reported at the Tanganyika Plantation Company (TPC) hospital’s malaria registers, meteorological data recorded at TPC sugar factory and data on bed nets distributed in Lower Moshi from 2009 to 2018 were studied. Correlation between bed nets distributed and malaria test positivity rates were explored by using Pearson correlation analysis and the associations between malaria test positivity rates and demographic and meteorological variables were determined by logistic regression and negative binomial regression analyses, respectively. Results Malaria cases reported at TPC hospital ranged between 0.48 and 2.26% per year and increased slightly at the introduction of malaria rapid diagnostic tests. The risk of testing positive for malaria were significantly highest among individuals aged between 6 and 15 years (OR = 1.65; 1.65 CI = 1.28–2.13; p = 0.001) and 16–30 years (OR = 1.49; CI = 1.17–1.89; p = 0.001) and when adjusted for age, the risk were significantly higher among male individuals when compared to female individuals (OR = 1.54; 1.00–1.31; p = 0.044). Malaria test positivity rates were positively associated with average monthly minimum temperatures and negatively associated with average monthly maximum temperatures (incidence rate ratio (IRR) = 1.37, 95% confidence interval (CI) = 1.05–1.78, p = 0.019 and IRR = 0.72, 95% CI = 0.58–0.91, p = 0.005, respectively). When analysed with one month lag for predictor variables, malaria test positivity rates were still significantly associated with average monthly minimum and maximum temperatures (IRR = 1.67, 95% CI = 1.28–2.19, p = 0.001 and IRR = 0.68, 95% CI = 0.54–0.85, p = 0.001, respectively). Average monthly rainfall and relative humidity with or without a one month lag was not associated with malaria test positivity rates in the adjusted models. Explopring possible associations between distribution of long-lasting insecticidal nets, (LLINs) and malaria test positivity rates showed no apparent correlation between numbers of LLINs distributed in a particular year and malaria test positivity rates. Conclusion In Lower Moshi, the risk of being tested positive for malaria was highest for older children and male individuals. Higher minimum and lower maximum temperatures were the strongest climatic predictors for malaria test positivity rates. In areas with extensive irrigation activity as in Lower Moshi, vector abundance and thus malaria transmission may be less dependent on rainfall patterns and humidity. Mass distribution of LLINs did not have an effect in this area with already very low malaria transmission.

Asymptomatic malaria and anemia among pregnant women during high and low malaria transmission seasons in Burkina Faso: household-based cross-sectional surveys in Burkina Faso, 2013–2017

Background: Malaria in endemic countries is often asymptomatic during pregnancy, but it has substantial consequences for both the mother and her unborn baby. In Burkina Faso, the prevalence and risk factors of asymptomatic malaria in pregnancy and its main consequence, anemia, during high and low malaria transmission seasons is underexplored at the household level.Methods: Data of 1751 pregnant women from October 2013 to March 2014 and 1931 pregnant women from April 2017 to June 2017 were drawn from two cross-sectional household surveys conducted in 24 health districts of Burkina Faso. Pregnant women were tested for malaria in their household after consenting. Asymptomatic carriage was defined as a positive result from malaria rapid diagnostic tests in the absence of malaria symptoms. Anemia was defined as hemoglobin level less than 11 g/dL.Results: Prevalence of asymptomatic malaria in pregnancy was estimated at 23.9% (95%CI: 20.2–28.0) during the high transmission season (October–November) in 2013. During the low transmission season, it was 12.7% (95%CI: 10.9–14.7) between December and March in 2013–2014 and halved (6.4%; 95%CI: 5.3–7.6) between April and June 2017. Anemia prevalence was estimated at 43.7% (95%CI: 39.3–48.3) during the high transmission season in 2013. During the low transmission season, it was 32.7% (95%CI: 30.1–35.5) between December and March 2013–2014 and 46.6% (95%CI: 44.3–48.9) between April and June, 2017.Conclusion: Prevalence of asymptomatic malaria in pregnancy was significantly higher during the high malaria transmission season while anemia prevalence was lower. Our study provides an opportunity to assess the prevalence of asymptomatic carriage and one of its main consequences, anemia, among pregnant women at the community level throughout the year. In order to mitigate the harmful effects of asymptomatic carriage for both the mother and fetus, health programs aimed at increasing the number of women coming into contact with health workers are necessary.

Molecular identification of native Wolbachia pipientis in Anopheles minimus in a low-malaria transmission area of Umphang Valley along the Thailand-Myanmar border

Background Wolbachia, obligate intracellular bacteria, infect the majority of arthropods, including many mosquito species of medical importance. Some Wolbachia strains interfere with the development of Plasmodium parasites in female Anopheles, a major vector of malaria. The use of Wolbachia as a means to block malaria transmission is an emerging vector control strategy in highly endemic areas. Hence, identification of native Wolbachia strains in areas where malaria transmission is low may uncover a particular Wolbachia strain capable of Plasmodium interference. This study aims to identify native Wolbachia strains in female Anopheles spp. that are predominant in a low-malaria transmission area in mainland Southeast Asia. Methods Following a 2-year survey of malaria vectors in Umphang Valley of Tak Province, Thailand, DNA extracts of female An. minimus, An. peditaeniatus, and An. maculatus were subjected to amplification of the conserved region of the 16S rRNA-encoding gene. The DNA sequences of the amplicons were phylogenetically compared with those of known Wolbachia strains. Results Among three Anopheles spp., amplification was detected in only the DNA samples from An. minimus. The DNA sequencing of amplicons revealed 100% similarity to Wolbachia pipientis, confirming the specificity of amplification. The Wolbachia-positive An. minimus samples were devoid of Plasmodium 18S rRNA amplification. The phylogenetic trees indicate a close relationship with Wolbachia strains in subgroup B. Conclusion To the best of our knowledge, the data presented herein provide the first molecular evidence of a Wolbachia strain in An. minimus, hereinafter named wAnmi, in a low-malaria transmission area in the Umphang Valley of western Thailand. Further biological characterization is required to examine its potential for malaria transmission control in the field.

High level of resistance in Anopheles arabiensis mosquito to pyrethroid insecticides from low malaria transmission zone of Moroto district, Karamoja region, Uganda: Implication for malaria vector control

Background: Karamoja region of Uganda previously classified as low malaria transmission zone is currently experiencing significant upsurge of malaria incidences. Long lasting insecticidal nets (LLINs) impregnated with pyrethroids constitute a major tool for malaria control in this region. Efficacy of this tool can be hampered by resistance to the pyrethroids in the Anopheles mosquito vectors. Resistance status of these mosquitoes in this region is poorly understood, effectively hampering better understanding of the impact of LLINs in the malaria control initiative. Here, we assessed susceptibility of the Anopheles arabiensis from the region to deltamethrin, permethrin (pyrethroids) and pirirmiphos-methyl (organophosphate) insecticides.Method: We collected anopheline mosquito larvae from their natural habitats and reared them to adult emergence in situ field insectary in Karamoja region. We then identified them morphological to species level and exposed 513 emerge adult female An gambiae s.l., mosquitoes to diagnostic dosages of deltamethrin (0.05%), permethrin (0.75%) and pirimiphos-methyl (0.25%) pyrethroids exposure using the standard WHO insecticide susceptibility test assay. Synergic assays using piperonyl butoxide (PBO) were done to check for the involvement of detoxification enzymes in pyrethroid resistant populations. We then screened for knockdown resistance (KDR) and mosquito species diversity using Polymerase Chain Reaction (PCR).Results: Majority (96%) of the mosquitoes we sampled were identified as An. arabiensis and 4% as An. gambiae sensu stricto. We observed cross-resistance to both deltamethrin (11.9%) and permethrin (47%) but susceptibility (100% mortality) to pirimiphos-methyl in An. arabiensis. The pre-exposure to PBO ameliorated the resistance to both pyrethroids. We detected homozygous KDR -eastern variant in 1.8 and 50% of the An. arabiensis and An. gambiae s.s. respectively.Conclusion: Anopheles arabiensis and An. gambiae s.s. are the malaria vector in Karamoja region with An. arabiensis predominating. Both species are susceptible to pirimiphos-methyl but resistant to both deltamethrin and permethrin, through a metabolic process (phenotype). Mosquotoes with genetic (kdr) mutations for resistance were minimal and hence have minimal contribution to the pyrethroid resistance profile. An. arabiensis can thus be controled in Karamoja region using deltamethrin and/or permethrin impregnated mosquito nets integrated with PBO and/or through indoor residual spraying of sprayable human dwellings with pirimiphos-methyl.