Stability of Dihydroartemisinin-Piperaquine Tablet Halves During Prolonged Storage Under Tropical Conditions.

Dihydroartemisinin-piperaquine (DP) is recommended for the treatment of uncomplicated malaria, used in efforts to contain artemisinin resistance, and increasingly considered for mass drug administration. Because of the narrow therapeutic dose range and available tablet strengths, the manufacturers and World Health Organization recommended regimens involve breaking tablets into halves to accurately dose children according to body weight. Use of tablet fractions in programmatic settings under tropical conditions requires a highly stable product; however, the stability of DP tablet fractions is unknown. We aged full and half DP (Eurartesim®) tablets in a stability chamber at 30°C and 70% humidity level. The active pharmaceutical ingredients dihydroartemisinin and piperaquine remained at ≥ 95% over the 3 months' period of ageing in light and darkness. These findings are reassuring for DP, but highlight the need to assess drug stability under real-life settings during the drug development process, particularly for key drugs of global disease control programs

Adaptive geostatistical design and analysis for prevalence surveys

Non-adaptive geostatistical designs (NAGDs) offer standard ways of collecting and analysing geostatistical data in which sampling locations are fixed in advance of any data collection. In contrast, adaptive geostatistical designs (AGDs) allow collection of geostatistical data over time to depend on information obtained from previous information to optimise data collection towards the analysis objective. AGDs are becoming more important in spatial mapping, particularly in poor resource settings where uniformly precise mapping may be unrealistically costly and the priority is often to identify critical areas where interventions can have the most health impact. Two constructions are: singleton and batch adaptive sampling. In singleton sampling, locations xi are chosen sequentially and at each stage, xk+1 depends on data obtained at locations x1,…,xk. In batch sampling, locations are chosen in batches of size b>1, allowing each new batch, {x(k+1),…,x(k+b)}, to depend on data obtained at locations x1,…,xkb. In most settings, batch sampling is more realistic than singleton sampling. We propose specific batch AGDs and assess their efficiency relative to their singleton adaptive and non-adaptive counterparts using simulations. We then show how we are applying these findings to inform an AGD of a rolling Malaria Indicator Survey, part of a large-scale, five-year malaria transmission reduction project in Malawi

Childhood malaria case incidence in Malawi between 2004 and 2017:Spatio-temporal modelling of climate and non-climate factors

Introduction Malaria transmission is influenced by a complex interplay of factors including climate, socio-economic, environmental factors and interventions. Malaria control efforts across Africa have shown a mixed impact. Climate driven factors may play an increasing role with climate change. Efforts to strengthen routine facility-based monthly malaria data collection across Africa create an increasingly valuable data source to interpret burden trends and monitor control programme progress. A better understanding of the association with other climatic and non-climatic drivers of malaria incidence over time and space may help guide and interpret the impact of interventions. Methods Routine monthly paediatric outpatient clinical malaria case data were compiled from 27 districts in Malawi between 2004 and 2017, and analysed in combination with data on climatic, environmental, socio-economic and interventional factors and district level population estimates. A spatio-temporal generalized linear mixed model was fitted using Bayesian inference, in order to quantify the strength of association of the various risk factors with district-level variation in clinical malaria rates in Malawi, and visualised using maps. Results Between 2004 and 2017 reported childhood clinical malaria case rates showed a slight increase, from 50 to 53 cases per 1000 population, with considerable variation across the country between climatic zones. Climatic and environmental factors, including average monthly air temperature and rainfall anomalies, normalized difference vegetative index (NDVI) and RDT use for diagnosis showed a significant relationship with malaria incidence. Temperature in the current month and in each of the 3 months prior showed a significant relationship with the disease incidence unlike rainfall anomaly which was associated with malaria incidence at only three months prior. Estimated risk maps show relatively high risk along the lake and Shire valley regions of Malawi. Conclusion Our modelling approach can identify locations likely to have unusually high or low risk of malaria incidence across Malawi, and distinguishes between contributions to risk that can be explained by measured risk-factors and unexplained residual spatial variation. Also, spatial statistical methods applied to readily available routine data provides an alternative information source that can supplement survey data in policy development and implementation to direct surveillance and intervention efforts

Inhibitory geostatistical designs for spatial prediction taking account of uncertain covariance structure

The problem of choosing spatial sampling designs for investigating an unobserved spatial phenomenon S arises in many contexts, for example, in identifying households to select for a prevalence survey to study disease burden and heterogeneity in a study region D. We studied randomized inhibitory spatial sampling designs to address the problem of spatial prediction while taking account of the need to estimate covariance structure. Two specific classes of design are inhibitory designs and inhibitory designs plus close pairs. In an inhibitory design, any pair of sample locations must be separated by at least an inhibition distance δ. In an inhibitory plus close pairs design, n − k sample locations in an inhibitory design with inhibition distance δ are augmented by k locations each positioned close to one of the randomly selected n − k locations in the inhibitory design, uniformly distributed within a disk of radius ζ. We present simulation results for the Matérn class of covariance structures. When the nugget variance is non-negligible, inhibitory plus close pairs designs demonstrate improved predictive efficiency over designs without close pairs. We illustrate how these findings can be applied to the design of a rolling Malaria Indicator Survey that forms part of an ongoing large-scale, 5-year malaria transmission reduction project in Malawi

Effects of transmission reduction by insecticide-treated bed nets (ITNs) on parasite genetics population structure: I. The genetic diversity of Plasmodium falciparum parasites by microsatellite markers in western Kenya

<p>Abstract</p> <p>Background</p> <p>Insecticide-treated bed nets (ITNs) reduce malaria transmission and are an important prevention tool. However, there are still information gaps on how the reduction in malaria transmission by ITNs affects parasite genetics population structure. This study examined the relationship between transmission reduction from ITN use and the population genetic diversity of <it>Plasmodium falciparum </it>in an area of high ITN coverage in western Kenya.</p> <p>Methods</p> <p>Parasite genetic diversity was assessed by scoring eight single copy neutral multilocus microsatellite (MS) markers in samples collected from <it>P. falciparum-</it>infected children (< five years) before introduction of ITNs (1996, baseline, n = 69) and five years after intervention (2001, follow-up, n = 74).</p> <p>Results</p> <p>There were no significant changes in overall high mixed infections and unbiased expected heterozygosity between baseline (%M_A= 94% and H_e= 0.75) and follow up (%M_A= 95% and H_e= 0.79) years. However, locus specific analysis detected significant differences for some individual loci between the two time points. Pfg377 loci, a gametocyte-specific MS marker showed significant increase in mixed infections and H_ein the follow up survey (%M_A= 53% and H_e= 0.57) compared to the baseline (%M_A= 30% and H_e= 0.29). An opposite trend was observed in the erythrocyte binding protein (EBP) MS marker. There was moderate genetic differentiation at the Pfg377 and TAA60 loci (F_ST= 0.117 and 0.137 respectively) between the baseline and post-ITN parasite populations. Further analysis revealed linkage disequilibrium (LD) of the microsatellites in the baseline (14 significant pair-wise tests and <it>I^S_A</it>= 0.016) that was broken in the follow up parasite population (6 significant pairs and <it>I^S_A</it>= 0.0003). The locus specific change in H_e, the moderate population differentiation and break in LD between the baseline and follow up years suggest an underlying change in population sub-structure despite the stability in the overall genetic diversity and multiple infection levels.</p> <p>Conclusions</p> <p>The results from this study suggest that although <it>P. falciparum </it>population maintained an overall stability in genetic diversity after five years of high ITN coverage, there was significant locus specific change associated with gametocytes, marking these for further investigation.</p

Effects of transmission reduction by insecticide-treated bed nets (ITNs) on parasite genetics population structure: I. The genetic diversity of Plasmodium falciparum parasites by microsatellite markers in western Kenya

<p>Abstract</p> <p>Background</p> <p>Insecticide-treated bed nets (ITNs) reduce malaria transmission and are an important prevention tool. However, there are still information gaps on how the reduction in malaria transmission by ITNs affects parasite genetics population structure. This study examined the relationship between transmission reduction from ITN use and the population genetic diversity of <it>Plasmodium falciparum </it>in an area of high ITN coverage in western Kenya.</p> <p>Methods</p> <p>Parasite genetic diversity was assessed by scoring eight single copy neutral multilocus microsatellite (MS) markers in samples collected from <it>P. falciparum-</it>infected children (< five years) before introduction of ITNs (1996, baseline, n = 69) and five years after intervention (2001, follow-up, n = 74).</p> <p>Results</p> <p>There were no significant changes in overall high mixed infections and unbiased expected heterozygosity between baseline (%M_A= 94% and H_e= 0.75) and follow up (%M_A= 95% and H_e= 0.79) years. However, locus specific analysis detected significant differences for some individual loci between the two time points. Pfg377 loci, a gametocyte-specific MS marker showed significant increase in mixed infections and H_ein the follow up survey (%M_A= 53% and H_e= 0.57) compared to the baseline (%M_A= 30% and H_e= 0.29). An opposite trend was observed in the erythrocyte binding protein (EBP) MS marker. There was moderate genetic differentiation at the Pfg377 and TAA60 loci (F_ST= 0.117 and 0.137 respectively) between the baseline and post-ITN parasite populations. Further analysis revealed linkage disequilibrium (LD) of the microsatellites in the baseline (14 significant pair-wise tests and <it>I^S_A</it>= 0.016) that was broken in the follow up parasite population (6 significant pairs and <it>I^S_A</it>= 0.0003). The locus specific change in H_e, the moderate population differentiation and break in LD between the baseline and follow up years suggest an underlying change in population sub-structure despite the stability in the overall genetic diversity and multiple infection levels.</p> <p>Conclusions</p> <p>The results from this study suggest that although <it>P. falciparum </it>population maintained an overall stability in genetic diversity after five years of high ITN coverage, there was significant locus specific change associated with gametocytes, marking these for further investigation.</p

Reducing contamination risk in cluster-randomized infectious disease-intervention trials

Background: Infectious disease interventions are increasingly tested using cluster-randomized trials (CRTs). These trial settings tend to involve a set of sampling units, such as villages, whose geographic arrangement may present a contamination risk in treatment exposure. The most widely used approach for reducing contamination in these settings is the so-called fried-egg design, which excludes the outer portion of all available clusters from the primary trial analysis. However, the fried-egg design ignores potential intra-cluster spatial heterogeneity and makes the outcome measure inherently less precise. Whereas the fried-egg design may be appropriate in specific settings, alternative methods to optimize the design of CRTs in other settings are lacking. Methods: We present a novel approach for CRT design that either fully includes or fully excludes available clusters in a defined study region, recognizing the potential for intra-cluster spatial heterogeneity. The approach includes an algorithm that allows investigators to identify the maximum number of clusters that could be included for a defined study region and maintain randomness in both the selection of included clusters and the allocation of clusters to either the treatment group or control group. The approach was applied to the design of a CRT testing the effectiveness of malaria vector-control interventions in southern Malawi. Conclusions: Those planning CRTs to evaluate interventions should consider the approach presented here during trial design. The approach provides a novel framework for reducing the risk of contamination among the CRT randomization units in settings where investigators determine the reduction of contamination risk as a high priority and where intra-cluster spatial heterogeneity is likely. By maintaining randomness in the allocation of clusters to either the treatment group or control group, the approach also permits a randomization-valid test of the primary trial hypothesis

Genetic diversity of Plasmodium falciparum parasite by microsatellite markers after scale-up of insecticide-treated bed nets in western Kenya

Background: An initial study of genetic diversity of Plasmodium falciparum in Asembo, western Kenya showed that the parasite maintained overall genetic stability 5 years after insecticide-treated bed net (ITN) introduction in 1997. This study investigates further the genetic diversity of P. falciparum 10 years after initial ITN introduction in the same study area and compares this with two other neighbouring areas, where ITNs were introduced in 1998 (Gem) and 2004 (Karemo). Methods: From a cross-sectional survey conducted in 2007, 235 smear-positive blood samples collected from children ≤15-year-old in the original study area and two comparison areas were genotyped employing eight neutral microsatellites. Differences in multiple infections, allele frequency, parasite genetic diversity and parasite population structure between the three areas were assessed. Further, molecular data reported previously (1996 and 2001) were compared to the 2007 results in the original study area Asembo. Results: Overall proportion of multiple infections (M A ) declined with time in the original study area Asembo (from 95.9 %-2001 to 87.7 %-2007). In the neighbouring areas, M A was lower in the site where ITNs were introduced in 1998 (Gem 83.7 %) compared to where they were introduced in 2004 (Karemo 96.7 %) in 2007. Overall mean allele count (M AC ~ 2.65) and overall unbiased heterozygosity (H e ~ 0.77) remained unchanged in 1996, 2001 and 2007 in Asembo and was the same level across the two neighbouring areas in 2007. Overall parasite population differentiation remained low over time and in the three areas at F ST < 0.04. Both pairwise and multilocus linkage disequilibrium showed limited to no significant association between alleles in Asembo (1996, 2001 and 2007) and between three areas. Conclusions: This study showed the P. falciparum high genetic diversity and parasite population resilience on samples collected 10 years apart and in different areas in western Kenya. The results highlight the need for long-term molecular monitoring after implementation and use of combined and intensive prevention and intervention measures in the region

Effect of Transmission Reduction by Insecticide-Treated Bednets (ITNs) on Antimalarial Drug Resistance in Western Kenya

Despite the clear public health benefit of insecticide-treated bednets (ITNs), the impact of malaria transmission-reduction by vector control on the spread of drug resistance is not well understood. In the present study, the effect of sustained transmission reduction by ITNs on the prevalence of Plasmodium falciparum gene mutations associated with resistance to the antimalarial drugs sulfadoxine-pyrimethamine (SP) and chloroquine (CQ) in children under the age of five years was investigated during an ITN trial in Asembo area, western Kenya. During the ITN trial, the national first line antimalarial treatment changed from CQ to SP. Smear-positive samples collected from cross sectional surveys prior to ITN introduction (baseline, n = 250) and five years post-ITN intervention (year 5 survey, n = 242) were genotyped for single nucleotide polymorphisms (SNPs) at dhfr-51, 59, 108, 164 and dhps-437, 540 (SP resistance), and pfcrt-76 and pfmdr1-86 (CQ resistance). The association between the drug resistance mutations and epidemiological variables was evaluated. There were significant increases in the prevalence of SP dhps mutations and the dhfr/dhps quintuple mutant, and a significant reduction in the proportion of mixed infections detected at dhfr-51, 59 and dhps-437, 540 SNPs from baseline to the year 5 survey. There was no change in the high prevalence of pfcrt-76 and pfmdr1-86 mutations. Multivariable regression analysis further showed that current antifolate use and year of survey were significantly associated with more SP drug resistance mutations. These results suggest that increased antifolate drug use due to drug policy change likely led to the high prevalence of SP mutations 5 years post-ITN intervention and reduced transmission had no apparent effect on the existing high prevalence of CQ mutations. There is no evidence from the current study that sustained transmission reduction by ITNs reduces the prevalence of genes associated with malaria drug resistance