Results of the first hydroacoustic survey of the Ugandan waters of Lake Victoria

A lakewide hydroacoustic research programme was designed in the Ugandan waters of Lake Victoria in order to ascertain the status of fish stocks. Data obtained from the hydroacoustic surveys were complemented with by catch data from multi mesh gillnets and frame trawls to validate acoustic estimates. Fish were distributed over the whole survey area, though the densities and species composition varied from place to place. Echo traces showed the fish formed schools during daytime and became more dispersed towards evening. Approximately equivalent indices of abundance were estimated for Rastrineobola argentea and Haplocromines. The distribution of the freshwater prawn, Caridina nilotica and the lakefly Chaoborus sp. was patchy. Dense swarms of Chaoborus larvae were observed to disperse from the lake bottom as the night approached thus assuring echo-traces formed by fish on the echogram and making their interpretation difficult. Caridina nilotica were observed to form dense echo-traces at the thermoclin

Uptake of trace elements by food crops grown within the Kilembe copper mine catchment, Western Uganda

The mining and processing of copper from the Kilembe mine between 1956 and 1982 left behind millions of tons of cupriferous and cobalt ferrous mine tailings within the Kilembe mine catchment. Subsequent erosion and deposition of the tailings into adjacent areas led to increased concentrations of Cu, Co, Ni, Zn, and Pb in the catchment soils. The Kilembe catchment is utilised for subsistence farming, producing mainly food crops, but there are also a number of settlements in the contaminated area. A study was conducted in 2016 to establish the concentrations of trace elements in a range of food crops grown within the catchment. Samples of maize, bananas, cassava, sweet potatoes, ground nuts, amaranthus, onions, beans and yams were collected, washed and oven dried at 80 °C. The dried foods were finely ground, microwave- digested in nitric acid and analysed using inductively coupled plasma mass spectrometry (ICPMS). All the foods grown in contaminated soils showed instances of higher concentrations of Cu, Co, Ni, Zn, and in some cases Pb, compared with controls grown in uncontaminated soils. Amaranthus accumulated a range of trace elements with 26% of the samples exceeding EC thresholds for Cu in vegetables of 26 mg kg−1.Other crops with elemental concentrations exceeding recommended thresholds in some of the samples included beans (Zn), yams (Zn and Pb) and ground nuts (Zn). The concentrations of trace elements in onions, cassava, sweet potatoes, bananas and maize were not significantly different from controls. However, strong and positive correlations between the trace elements were found in beans, yams, amaranthus, maize and ground nuts, suggesting a common source of trace metals. There was strong evidence of soil dust retention on leaf vegetables (Amaranthus) despite washing. The accumulation of trace elements in the edible parts of vegetables and foods could have a direct impact on the health of local people, because the foods produced from gardens are mostly consumed locally

Drug resistance mediating Plasmodium falciparum polymorphisms and clinical presentations of parasitaemic children in Uganda.

BackgroundPlasmodium falciparum genetic polymorphisms that mediate altered drug sensitivity may impact upon virulence. In a cross-sectional study, Ugandan children with infections mutant at pfcrt K76T, pfmdr1 N86Y, or pfmdr1 D1246Y had about one-fourth the odds of symptomatic malaria compared to those with infections with wild type (WT) sequences. However, results may have been confounded by greater likelihood in those with symptomatic disease of higher density mixed infections and/or recent prior treatment that selected for WT alleles.MethodsPolymorphisms in samples from paired episodes of asymptomatic and symptomatic parasitaemia in 114 subjects aged 4-11 years were followed longitudinally in Tororo District, Uganda. Paired episodes occurred within 3-12 months of each other and had no treatment for malaria in the prior 60 days. The prevalence of WT, mixed, and mutant alleles was determined using multiplex ligase detection reaction-fluorescent microsphere assays.ResultsConsidering paired episodes in the same subject, the odds of symptomatic malaria were lower for infections with mutant compared to WT or mixed sequence at N86Y (OR 0.26, 95% CI 0.09-0.79, p = 0.018), but not K76T or D1246Y. However, symptomatic episodes (which had higher densities) were more likely than asymptomatic to be mixed (for N86Y OR 2.0, 95% CI 1.04-4.0, p = 0.036). Excluding mixed infections, the odds of symptomatic malaria were lower for infections with mutant compared to WT sequence at N86Y (OR 0.33, 95% CI 0.11-0.98, p = 0.046), but not the other alleles. However, if mixed genotypes were grouped with mutants in this analysis or assuming that mixed infections consisted of 50% WT and 50% mutant genotypes, the odds of symptomatic infection did not differ between infections that were mutant or WT at the studied alleles.ConclusionsAlthough infections with only the mutant pfmdr1 86Y genotype were associated with symptomatic infection, this association could primarily be explained by greater parasite densities and therefore greater prevalence of mixed infections in symptomatic children. These results indicate limited association between the tested polymorphisms and risk of symptomatic disease and highlight the value of longitudinal studies for assessing associations between parasite factors and clinical outcomes

Trench layering using indole-3-butyric acid and local organic substrate mixtures to enhance rooting and survival of apple rootstocks

Apples ( Malus domestica ) were introduced to enhance nutrition and improve livelihoods of famers in highlands of Uganda. However, adoption and commercialisation of apples is largely constrained by low quality rootstocks due to poor rooting and low survivability. This study determined the effect of organic substrate mixtures (OSM) and indole-3-butyric acid (IBA) on rooting, sprouting and survival characteristics of apple rootstocks propagated by trench layering. Four apple rootstock varieties (M106, M109, MM793 and bitten-felder) were planted in OSM (Site soil as control, TsaOm and TsdOm) then treated with IBA concentrations (IBA-0 ppm, IBA-4000 ppm and IBA-8000 ppm) in a factorial randomised block design with three replications. Results showed significant (P<0.001) variability among rootstocks for all characters. OSM significantly (P<0.01) increased rooting, root numbers and root length while IBA significantly (P<0.01) increased all traits, except rooting. The highest rooting (46.7%), root numbers (23.1) and root length (14.9cm), and sprout length (59.5cm) were obtained in bitten felder under TsaOm + IBA-4000ppm, TsaOm, and TsdOm, respectively. For M106, maximum rooting (28.1%) and root numbers (22.3) were obtained under TsaOm + IBA-8000ppm while root (14.3cm) and sprout (35.2cm) lengths under TsdOm. TsaOm gave the highest root numbers (14.2), root (12.8cm) and sprout (30.7cm) lengths in M109 likewise root (7.8cm) and sprout (38.3cm) lengths in MM793. Logistic regression revealed that rooting, sprout length, and IBA-4000ppm significantly (P<0.01) increased survival of apple rootstocks. The highest survival rates in bitten felder and M106 were 52.4% and 51.7% under TsdOm + IBA-4000 ppm and TsdOm + IBA-8000 ppm respectively; likewise 49.5% in M109 and 51.7% in MM793 both treated with IBA-8000 ppm. The results demonstrate that trench layering with OSM and IBA improves rooting and survival of apple rootstocks which might improve farmers\u2019 access to quality apple planting material.Les pommes ( Malus domestica ) \ue9taient introduites pour renforcer la nutrition et am\ue9liorer le niveau de vie des producteurs dans les r\ue9gions montagneuses d\u2019Ouganda. Toutefois, l\u2019adoption et la commercialisation des pommes sont largement limit\ue9es par la faible qualit\ue9 des porte-greffes \ue0 cause du pauvre enracinement et la faible capacit\ue9 de survie. Cette \ue9tude visait \ue0 d\ue9terminer l\u2019effet des m\ue9langes du substrat organique (OSM) et l\u2019acide indole-3-butyrique (IBA) sur l\u2019enracinement, la germination et les caract\ue9ristiques de survie des porte-greffes de la pomme propag\ue9e par le marcottage de tranch\ue9e. Quatre portes greffes des vari\ue9t\ue9s de pomme (M106, M109, MM793 et bitten-felder) \ue9taient plant\ue9es dans l\u2019OSM (site de sol servant de contr\uf4le, TsaOm et TSdOm) et trait\ue9e avec diff\ue9rentes concentrations de l\u2019IBA (IBA-0 ppm, IBA-4000 ppm et IBA-8000 ppm) dans un design factoriel \ue0 blocks compl\ue8tement al\ue9atoires avec trois r\ue9plications. Les r\ue9sultats ont montr\ue9 de variabilit\ue9 significative (P<0,001) entre les porte-greffes des vari\ue9t\ue9s pour tous les caract\ue8res. OSM significativement (P<0.01) a fait accro\ueetre l\u2019enracinement, le nombre de racines et la longueur de la racine; alors que IBA a augment\ue9 significativement (P<0.001) tous les traits, sauf l\u2019enracinement. Les plus grandes valeurs des caract\ue8res \ue0 savoir\ua0; l\u2019enracinement (46,7%), le nombre de racines (23,1), la longueur des racines (14,9 cm), et la longueur des pousses (59,5cm) \ue9taient obtenues sur bitten felder sous TsaOm + IBA-4000ppm, TsaOm, et TsdOm, respectivement. Pour M106, les valeurs maximales de l\u2019enracinement (28,1%) et de nombre de racines (22.3) \ue9taient obtenues sous TsaOm + IBA-8000ppm alors que les valeurs maximales de la longueur des racines (14,3 cm) et des pousses (35,2 cm) sous TsdOM. TsaOm ont donn\ue9 le plus grand nombre de racines (14,2), la plus grande longueur des racines (12,8 cm) et de pousses (30,7 cm) dans M109 pareillement \ue0 la longueur des racines (7,8 cm) et de pousses (38,3 cm) dans MM793. La r\ue9gression logistique a r\ue9v\ue9l\ue9 que l\u2019enracinement, la longueur de la pousse et IBA-400ppm ont augment\ue9 significativement (P<0,01) la survie des porte-greffes de la pomme. Les plus forts taux de survies sur le bitten felder et M106 \ue9taient 52,4% et 51,7% sous TsdOm+IBA-4000 ppm et TsdOm+IBA-8000 ppm, respectivement\ua0; de m\ueame 49,5% ont \ue9t\ue9 obtenus sur M109 et 51,7% sur MM793 tous trait\ue9s avec IBA-8000 ppm. Les r\ue9sultats ont d\ue9montr\ue9 que le marcottage de tranch\ue9e avec OSM et IBA am\ue9liore l\u2019racinement et la survie des porte-greffes qui pourrait am\ue9liorer l\u2019acc\ue8s des producteurs \ue0 des mat\ue9riels de plantation de pomme de qualit\ue9

Drug susceptibility of Plasmodium falciparum in eastern Uganda: a longitudinal phenotypic and genotypic study

Background: Treatment and control of malaria depends on artemisinin-based combination therapies (ACTs) and is challenged by drug resistance, but thus far resistance to artemisinins and partner drugs has primarily occurred in southeast Asia. The aim of this study was to characterise antimalarial drug susceptibility of Plasmodium falciparum isolates from Tororo and Busia districts in Uganda. Methods: In this prospective longitudinal study, P falciparum isolates were collected from patients aged 6 months or older presenting at the Tororo District Hospital (Tororo district, a site with relatively low malaria incidence) or Masafu General Hospital (Busia district, a high-incidence site) in eastern Uganda with clinical symptoms of malaria, a positive Giemsa-stained blood film for P falciparum, and no signs of severe disease. Ex-vivo susceptibilities to ten antimalarial drugs were measured using a 72-h microplate growth inhibition assay with SYBR Green detection. Relevant P falciparum genetic polymorphisms were characterised by molecular methods. We compared results with those from earlier studies in this region and searched for associations between drug susceptibility and parasite genotypes. Findings: From June 10, 2016, to July 29, 2019, 361 P falciparum isolates were collected in the Busia district and 79 in the Tororo district from 440 participants. Of 440 total isolates, 392 (89%) successfully grew in culture and showed excellent drug susceptibility for chloroquine (median half-maximal inhibitory concentration [IC50] 20·0 nM [IQR 12·0-26·0]), monodesethylamodiaquine (7·1 nM [4·3-8·9]), pyronaridine (1·1 nM [0·7-2·3]), piperaquine (5·6 nM [3·3-8·6]), ferroquine (1·8 nM [1·5-3·3]), AQ-13 (24·0 nM [17·0-32·0]), lumefantrine (5·1 nM [3·2-7·7]), mefloquine (9·5 nM [6·6-13·0]), dihydroartemisinin (1·5 nM [1·0-2·0]), and atovaquone (0·3 nM [0·2-0·4]). Compared with results from our study in 2010-13, significant improvements in susceptibility were seen for chloroquine (median IC50 288·0 nM [IQR 122·0-607·0]; p\u3c0·0001), monodesethylamodiaquine (76·0 nM [44·0-137]; p\u3c0·0001), and piperaquine (21·0 nM [7·6-43·0]; p\u3c0·0001), a small but significant decrease in susceptibility was seen for lumefantrine (3·0 nM [1·1-7·6]; p\u3c0·0001), and no change in susceptibility was seen with dihydroartemisinin (1·3 nM [0·8-2·5]; p=0·64). Chloroquine resistance (IC50\u3e100 nM) was more common in isolates from the Tororo district (11 [15%] of 71), compared with those from the Busia district (12 [4%] of 320; p=0·0017). We showed significant increases between 2010-12 and 2016-19 in the prevalences of wild-type P falciparum multidrug resistance protein 1 (PfMDR1) Asn86Tyr from 60% (391 of 653) to 99% (418 of 422; p\u3c0·0001), PfMDR1 Asp1246Tyr from 60% (390 of 650) to 90% (371 of 419; p\u3c0·0001), and P falciparum chloroquine resistance transporter (PfCRT) Lys76Thr from 7% (44 of 675) to 87% (364 of 417; p\u3c0·0001). Interpretation: Our results show marked changes in P falciparum drug susceptibility phenotypes and genotypes in Uganda during the past decade. These results suggest that additional changes will be seen over time and continued surveillance of susceptibility to key ACT components is warranted. Funding: National Institutes of Health and Medicines for Malaria Venture

Associations between Varied Susceptibilities to PfATP4 Inhibitors and Genotypes in Ugandan Plasmodium falciparum Isolates.

Among novel compounds under recent investigation as potential new antimalarial drugs are three independently developed inhibitors of the Plasmodium falciparum P-type ATPase (PfATP4): KAE609 (cipargamin), PA92, and SJ733. We assessed ex vivo susceptibilities to these compounds of 374 fresh P. falciparum isolates collected in Tororo and Busia districts, Uganda, from 2016 to 2019. Median IC50s were 65 nM for SJ733, 9.1 nM for PA92, and 0.5 nM for KAE609. Sequencing of pfatp4 for 218 of these isolates demonstrated many nonsynonymous single nucleotide polymorphisms; the most frequent mutations were G1128R (69% of isolates mixed or mutant), Q1081K/R (68%), G223S (25%), N1045K (16%), and D1116G/N/Y (16%). The G223S mutation was associated with decreased susceptibility to SJ733, PA92, and KAE609. The D1116G/N/Y mutations were associated with decreased susceptibility to SJ733, and the presence of mutations at both codons 223 and 1116 was associated with decreased susceptibility to PA92 and SJ733. In all of these cases, absolute differences in susceptibilities of wild-type (WT) and mutant parasites were modest. Analysis of clones separated from mixed field isolates consistently identified mutant clones as less susceptible than WT. Analysis of isolates from other sites demonstrated the presence of the G223S and D1116G/N/Y mutations across Uganda. Our results indicate that malaria parasites circulating in Uganda have a number of polymorphisms in PfATP4 and that modestly decreased susceptibility to PfATP4 inhibitors is associated with some mutations now present in Ugandan parasites

Impact of Antimalarial Treatment and Chemoprevention on the Drug Sensitivity of Malaria Parasites Isolated from Ugandan Children

Changing treatment practices may be selecting for changes in the drug sensitivity of malaria parasites. We characterized ex vivo drug sensitivity and parasite polymorphisms associated with sensitivity in 459 Plasmodium falciparum samples obtained from subjects enrolled in two clinical trials in Tororo, Uganda, from 2010 to 2013. Sensitivities to chloroquine and monodesethylamodiaquine varied widely; sensitivities to quinine, dihydroartemisinin, lumefantrine, and piperaquine were generally good. Associations between ex vivo drug sensitivity and parasite polymorphisms included decreased chloroquine and monodesethylamodiaquine sensitivity and increased lumefantrine and piperaquine sensitivity with pfcrt 76T, as well as increased lumefantrine sensitivity with pfmdr1 86Y, Y184, and 1246Y. Over time, ex vivo sensitivity decreased for lumefantrine and piperaquine and increased for chloroquine, the prevalences of pfcrt K76 and pfmdr1 N86 and D1246 increased, and the prevalences of pfdhfr and pfdhps polymorphisms associated with antifolate resistance were unchanged. In recurrent infections, recent prior treatment with artemether-lumefantrine was associated with decreased ex vivo lumefantrine sensitivity and increased prevalence of pfcrt K76 and pfmdr1 N86, 184F, and D1246. In children assigned chemoprevention with monthly dihydroartemisinin-piperaquine with documented circulating piperaquine, breakthrough infections had increased the prevalence of pfmdr1 86Y and 1246Y compared to untreated controls. The noted impacts of therapy and chemoprevention on parasite polymorphisms remained significant in multivariate analysis correcting for calendar time. Overall, changes in parasite sensitivity were consistent with altered selective pressures due to changing treatment practices in Uganda. These changes may threaten the antimalarial treatment and preventive efficacies of artemether-lumefantrine and dihydroartemisinin-piperaquine, respectively

Artemether-Lumefantrine to treat Malaria in pregnancy is associated with reduced placental Haemozoin deposition compared to Quinine in a randomized controlled trial

Data on efficacy of artemisinin-based combination therapy (ACT) to treat Plasmodium falciparum during pregnancy in sub-Saharan Africa is scarce. A recent open label, randomized controlled trial in Mbarara, Uganda demonstrated that artemether-lumefantrine (AL) is not inferior to quinine to treat uncomplicated malaria in pregnancy. Haemozoin can persist in the placenta following clearance of parasites, however there is no data whether ACT can influence the amount of haemozoin or the dynamics of haemozoin clearance

Population exposure to trace elements in the Kilembe copper mine area, Western Uganda: a pilot study

The mining and processing of copper in Kilembe, Western Uganda, from 1956 to 1982 left over 15 Mt. of tailings containing cupriferous and cobaltiferous pyrite dumped within a mountain river valley. This pilot study was conducted to assess the nature and extent of risk to local populations from metal contamination arising from those mining activities. We determined trace element concentrations in mine tailings, soils, locally cultivated foods,house dust, drinking water and human biomarkers (toenails) using ICP-MS analysis of acid digested samples. The results showed that tailings, containing higher concentrations of Co, Cu, Ni and As compared with world average crust values had eroded and contaminated local soils. Pollution load indices revealed that 51% of agricultural soils sampled were contaminated with trace elements. Local water supplies were contaminated, with Co concentrations that exceeded Wisconsin (US) thresholds in 25% of domestic water supplies and 40% of Nyamwamba river water samples. Zinc exceeded WHO/FAO thresholds of 99.4 mg kg−1 in 36% of Amaranthus vegetable samples, Cu exceeded EC thresholds of 20 mg kg−1 in 19% of Amaranthus while Pb exceeded WHO thresholds of 0.3 mg kg−1 in 47% of Amaranthus vegetables. In bananas, 20% of samples contained Pb concentrations that exceeded the WHO/FAO recommended threshold of 0.3 mg kg−1. However, risk assessment of local foods and water, based on hazard quotients (HQ values) revealed no potential health effects. The high external contamination of volunteers' toenails with some elements (even after a washing process) calls into question their use as a biomarker for metal exposure in human populations where feet are frequently exposed to soil

Preclinical characterization and target validation of the antimalarial pantothenamide MMV693183

Drug resistance and a dire lack of transmission-blocking antimalarials hamper malaria elimination. Here, we present the pantothenamide MMV693183 as a first-in-class acetyl-CoA synthetase (AcAS) inhibitor to enter preclinical development. Our studies demonstrate attractive drug-like properties and in vivo efficacy in a humanized mouse model of Plasmodium falciparum infection. The compound shows single digit nanomolar in vitro activity against P. falciparum and P. vivax clinical isolates, and potently blocks P. falciparum transmission to Anopheles mosquitoes. Genetic and biochemical studies identify AcAS as the target of the MMV693183-derived antimetabolite, CoA-MMV693183. Pharmacokinetic-pharmacodynamic modelling predict that a single 30 mg oral dose is sufficient to cure a malaria infection in humans. Toxicology studies in rats indicate a > 30-fold safety margin in relation to the predicted human efficacious exposure. In conclusion, MMV693183 represents a promising candidate for further (pre)clinical development with a novel mode of action for treatment of malaria and blocking transmission