Pooled-DNA sequencing identifies genomic regions of selection in Nigerian isolates of Plasmodium falciparum.

BACKGROUND: The burden of falciparum malaria is especially high in sub-Saharan Africa. Differences in pressure from host immunity and antimalarial drugs lead to adaptive changes responsible for high level of genetic variations within and between the parasite populations. Population-specific genetic studies to survey for genes under positive or balancing selection resulting from drug pressure or host immunity will allow for refinement of interventions. METHODS: We performed a pooled sequencing (pool-seq) of the genomes of 100 Plasmodium falciparum isolates from Nigeria. We explored allele-frequency based neutrality test (Tajima's D) and integrated haplotype score (iHS) to identify genes under selection. RESULTS: Fourteen shared iHS regions that had at least 2 SNPs with a score > 2.5 were identified. These regions code for genes that were likely to have been under strong directional selection. Two of these genes were the chloroquine resistance transporter (CRT) on chromosome 7 and the multidrug resistance 1 (MDR1) on chromosome 5. There was a weak signature of selection in the dihydrofolate reductase (DHFR) gene on chromosome 4 and MDR5 genes on chromosome 13, with only 2 and 3 SNPs respectively identified within the iHS window. We observed strong selection pressure attributable to continued chloroquine and sulfadoxine-pyrimethamine use despite their official proscription for the treatment of uncomplicated malaria. There was also a major selective sweep on chromosome 6 which had 32 SNPs within the shared iHS region. Tajima's D of circumsporozoite protein (CSP), erythrocyte-binding antigen (EBA-175), merozoite surface proteins - MSP3 and MSP7, merozoite surface protein duffy binding-like (MSPDBL2) and serine repeat antigen (SERA-5) were 1.38, 1.29, 0.73, 0.84 and 0.21, respectively. CONCLUSION: We have demonstrated the use of pool-seq to understand genomic patterns of selection and variability in P. falciparum from Nigeria, which bears the highest burden of infections. This investigation identified known genomic signatures of selection from drug pressure and host immunity. This is evidence that P. falciparum populations explore common adaptive strategies that can be targeted for the development of new interventions

Status of insecticide susceptibility in Anopheles gambiae s.l. from malaria surveillance sites in The Gambia

BACKGROUND: Vector control is an effective way of reducing malaria transmission. The main vector control methods include the use of insecticide-treated bed nets and indoor residual spraying (IRS). Both interventions rely on the continuing susceptibility of Anopheles to a limited number of insecticides. However, insecticide resistance, in particular pyrethroid-DDT cross-resistance, is a challenge facing malaria vector control in Africa because pyrethroids represent the only class of insecticides approved for treating bed nets and DDT is commonly used for IRS. Here baseline data are presented on the insecticide susceptibility levels of malaria vectors prior to The Gambian indoor residual spraying intervention programme. METHODS: Anopheles larvae were collected from six malaria surveillance sites (Brikama, Essau, Farafenni, Mansakonko, Kuntaur and Basse) established by the National Malaria Control Programme and the UK Medical Research Council Laboratories in The Gambia. The mosquitoes were reared to adulthood and identified using morphological keys and a species-specific polymerase chain reaction assay. Two- to three-day old adult female mosquitoes were tested for susceptibility to permethrin, deltamethrin and DDT using standard WHO protocols, insecticide susceptibility test kits and treated papers. RESULTS: All Anopheles mosquitoes tested belonged to the Anopheles gambiae complex. Anopheles arabiensis was predominant (54.1%), followed by An. gambiae s.s. (26.1%) and Anopheles melas (19.8%). Anopheles gambiae s.s. and An. arabiensis were found at all six sites. Anopheles melas was recorded only at Brikama. Mosquitoes from two of the six sites (Brikama and Basse) were fully susceptible to all three insecticides tested. However, DDT resistance was found in An. gambiae from Essau where the 24 hours post-exposure mortality was <80% but 88% for permethrin and 92% for deltamethrin. CONCLUSION: This current survey of insecticide resistance in Anopheles provides baseline information for monitoring resistance in The Gambia and highlights the need for routine resistance surveillance as an integral part of the proposed nation wide IRS intervention using DDT

Comparative Evaluation of Antioxidant Activity and Phytochemical Profile of Four Capsicum Fruits Species

Capsicum fruits are commonly consumed vegetables included in foods worldwide as spices. Capsicum species are known to improve digestion and appetite, cure colds, coughs, fever, colic, dysentery, worms, and piles among others. This research is therefore aimed at evaluating the phytochemicalas well as the antioxidant activities of four different varieties of Capsicum commonly consumed in Africa. The fruits were extracted using 3 different solvents; n-hexane, ethyl acetate, and methanol. The phytochemical evaluation was carried out using standard methods. Antioxidant activities were evaluated using 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. The highest yield of extract was obtained with methanol from Capsicum annuum var. grossum (25.48%); The phytochemical screening of crude extracts revealed the presence of alkaloids, saponins, flavonoids, phenols, terpenoids, tannins, coumarins, and cardiac glycosides. The total phenolic contents of crude extracts of Capsicum species ranged from 25.10±0.3 to 62.01±0.1 mg GAE g-1 (Gallic acid equivalent) of dry weight and the flavonoid contents ranged from 206.08±0.03 to 474.23±0.04 mg QE g-1 (Quercetin equivalent) of dry weight. The capacity to neutralize DPPH radicals was found to be at the highest in methanolic extracts of Capsicum species which neutralized 50% of free radicals at the concentrations of 5.79 μg mL-1, 8.08 μg mL-1, 5.76 μg mL-1, and 8.81 μg mL-1for Capsicum annuum var. accuminatum, Capsicum frutescens var. baccatum, Capsicum annuumvar. grossum and Capsicum annuum var. abbreviatum respectively. The study has concluded that Capsicum species contain phytoconstituents with high antioxidant activities and great potential to combat oxidative stress and other related diseases

Effective Doses and Excess Lifetime Cancer Risks from Absorbed Dose Rates Measured in Facilities of Two Tertiary Institutions in Nigeria

This study was aimed at examining the radiation absorbed dose rate, annual effective doses and excess lifetime cancer risks of halls of residence, Radiotherapy Unit and Radiology Department of UI, UCH and UNIMEDTH respectively. Results of measurements show that the mean absorbed dose rate for male and female hall are 0.33 ± 0.05476 and 0.17 ± 0.05074 µSv h-1 respectively. The mean overall absorbed dose rates calculated for facilities studied are 0.269 ± 0.0992 µSv h-1, 0.121 ± 0.036 µSv h-1 and 0.123 ± 0.00931 µSv h-1 in UI, UCH and UNIMEDTH respectively. The mean annual effective doses recorded in both male and female halls in University of Ibadan ranges between 0.71 mSv y-1 and 2.67 mSv y-1. The mean annual effective doses obtained from the facility of University of Medical Sciences Teaching Hospital, Ondo (UNIMEDTH) ranges between 0.17 and 0.44 mSv y-1. In addition, the mean annual effective doses calculated from the measured absorbed dose rate in Radiotherapy Department of University College Hospital, Ibadan ranges between 0.20 and 1.22 mSv y-1. As regards ELCRs, the mean values &nbsp;determined in various facilities examined are 6.07 x 10-3 (Male Halls, UI), 3.27 x 10-3 (Female Halls, UI), 0.57 x 10-3 (UNIMEDTH- NE), 0.99 x 10-3 (UNIMEDTH- EX), 0.65 x x 10-3 (Teletherapy, UCH) and 0.57 x 10-3 (Brachytherapy, UCH).The mean ELCRs of both halls examined are higher than the world average of 1.45 x 10-3 and the standard value of 0.29 x 10-3 by at least a factor of 1.97 units

Molecular Identification And Population Dynamics Of The Major Malaria Vectors In A Rainforest Zone Of Nigeria

Adult female mosquito vectors were collected from three villages in a typical rain forest area of Nigeria where no information exists on the major malaria vectors associated with human malaria. Sampling was carried out between January 2004 and January 2005 using pyrethrum and Human landing catch (HLC) techniques. A total catch of 2010 mosquitoes was recorded out of which 1800 were morphologically identified as female Anopheline mosquitoes. Further identification of the Anopheline species using the morphological keys revealed that 1399 (77.7%) belonged to the Anopheles gambiae s.l. and 401 (22.3%) to Anopheles funestus . A PCR based test on the Anopheles gambiae group identified 636 (45.5%) as Anopheles gambiae s.s and 763 (54.5%) as Anopheles arabiensis respectively. The cocktail PCR-assayon the total Anopheles funestus group showed 307 (76.6%), to be Anopheles funestus s.s and 94 (23.4%) to be Anopheles leesoni . The total number of Anopheles gambiae collected across the 3 villages was significantly higher (P&lt;0.05) than the number of Anopheles funestus caught within the same period. However, there was a seasonal difference in the population of Anopheline species collected in which the wet season collections constitute 45.4% An.gambiae s.l and 17.7% An.funestus while the dry season population constitutes 32.3% An.gambiae s.l and 4.5% An.funestus. The dry seasoncollections were predominantly An. arabiensis producing 23.9% of the total catch in.The overall number of Anopheles mosquitoes collected in the wet season was significantly higher than that of the dry season (P&lt;0.01). Generally, low sporozoite rates were recorded in all the communities and this may be an indication that transmission in this area is less intense. This study provides information on mosquito ecology, genetic and molecular techniques for identification of species complexes which are important strategies for planning malaria control programmes

Using a New Odour-Baited Device to Explore Options for Luring and Killing Outdoor-Biting Malaria Vectors: A Report on Design and Field Evaluation of the Mosquito Landing Box.

Mosquitoes that bite people outdoors can sustain malaria transmission even where effective indoor interventions such as bednets or indoor residual spraying are already widely used. Outdoor tools may therefore complement current indoor measures and improve control. We developed and evaluated a prototype mosquito control device, the 'Mosquito Landing Box' (MLB), which is baited with human odours and treated with mosquitocidal agents. The findings are used to explore technical options and challenges relevant to luring and killing outdoor-biting malaria vectors in endemic settings. Field experiments were conducted in Tanzania to assess if wild host-seeking mosquitoes 1) visited the MLBs, 2) stayed long or left shortly after arrival at the device, 3) visited the devices at times when humans were also outdoors, and 4) could be killed by contaminants applied on the devices. Odours suctioned from volunteer-occupied tents were also evaluated as a potential low-cost bait, by comparing baited and unbaited MLBs. There were significantly more Anopheles arabiensis, An. funestus, Culex and Mansonia mosquitoes visiting baited MLB than unbaited controls (P<=0.028). Increasing sampling frequency from every 120 min to 60 and 30 min led to an increase in vector catches of up to 3.6 fold (P<=0.002), indicating that many mosquitoes visited the device but left shortly afterwards. Outdoor host-seeking activity of malaria vectors peaked between 7:30 and 10:30pm, and between 4:30 and 6:00am, matching durations when locals were also outdoors. Maximum mortality of mosquitoes visiting MLBs sprayed or painted with formulations of candidate mosquitocidal agent (pirimiphos-methyl) was 51%. Odours from volunteer occupied tents attracted significantly more mosquitoes to MLBs than controls (P<0.001). While odour-baited devices such as the MLBs clearly have potential against outdoor-biting mosquitoes in communities where LLINs are used, candidate contaminants must be those that are effective at ultra-low doses even after short contact periods, since important vector species such as An. arabiensis make only brief visits to such devices. Natural human odours suctioned from occupied dwellings could constitute affordable sources of attractants to supplement odour baits for the devices. The killing agents used should be environmentally safe, long lasting, and have different modes of action (other than pyrethroids as used on LLINs), to curb the risk of physiological insecticide resistance

Anopheles gambiae distribution and insecticide resistance in the cities of Douala and Yaoundé (Cameroon): influence of urban agriculture and pollution

Background: Urban malaria is becoming a major health priority across Africa. A study was undertaken to assess the importance of urban pollution and agriculture practice on the distribution and susceptibility to insecticide of malaria vectors in the two main cities in Cameroon. Methods: Anopheline larval breeding sites were surveyed and water samples analysed monthly from October 2009 to December 2010. Parameters analysed included turbidity, pH, temperature, conductivity, sulfates, phosphates,nitrates, nitrites, ammonia, aluminium, alkalinity, iron, potassium, manganese, magnesium, magnesium hardness and total hardness. Characteristics of water bodies in urban areas were compared to rural areas and between urban sites. The level of susceptibility of Anopheles gambiae to 4% DDT, 0.75% permethrin, 0.05% deltamethrin, 0.1% bendiocarb and 5% malathion were compared between mosquitoes collected from polluted, non polluted and cultivated areas. Results: A total of 1,546 breeding sites, 690 in Yaoundé and 856 in Douala, were sampled in the course of the study. Almost all measured parameters had a concentration of 2- to 100-fold higher in urban compare to rural breeding sites. No resistance to malathion was detected, but bendiocarb resistance was present in Yaounde. Very low mortality rates were observed following DDT or permethrin exposure, associated with high kdr frequencies. Mosquitoes collected in cultivated areas, exhibited the highest resistant levels. There was little difference in insecticide resistance or kdr allele frequency in mosquitoes collected from polluted versus non-polluted sites. Conclusion: The data confirm high selection pressure on mosquitoes originating from urban areas and suggest urban agriculture rather than pollution as the major factor driving resistance to insecticide

Impact of urban agriculture on malaria vectors in Accra, Ghana

To investigate the impact of urban agriculture on malaria transmission risk in urban Accra larval and adult stage mosquito surveys, were performed. Local transmission was implicated as Anopheles spp. were found breeding and infected Anopheles mosquitoes were found resting in houses in the study sites. The predominant Anopheles species was Anopheles gambiae s.s.. The relative proportion of molecular forms within a subset of specimens was 86% S-form and 14% M-form. Anopheles spp. and Culex quinquefasciatus outdoor biting rates were respectively three and four times higher in areas around agricultural sites (UA) than in areas far from agriculture (U). The annual Entomological Inoculation Rate (EIR), the number of infectious bites received per individual per year, was 19.2 and 6.6 in UA and U sites, respectively. Breeding sites were highly transitory in nature, which poses a challenge for larval control in this setting. The data also suggest that the epidemiological importance of urban agricultural areas may be the provision of resting sites for adults rather than an increased number of larval habitats. Host-seeking activity peaked between 2–3 am, indicating that insecticide-treated bednets should be an effective control method

The Impact of Pyrethroid Resistance on the Efficacy of Insecticide-Treated Bed Nets against African Anopheline Mosquitoes: Systematic Review and Meta-Analysis

Background Pyrethroid insecticide-treated bed nets (ITNs) help contribute to reducing malaria deaths in Africa, but their efficacy is threatened by insecticide resistance in some malaria mosquito vectors. We therefore assessed the evidence that resistance is attenuating the effect of ITNs on entomological outcomes. Methods and Findings We included laboratory and field studies of African malaria vectors that measured resistance at the time of the study and used World Health Organization–recommended impregnation regimens. We reported mosquito mortality, blood feeding, induced exophily (premature exit of mosquitoes from the hut), deterrence, time to 50% or 95% knock-down, and percentage knock-down at 60 min. Publications were searched from 1 January 1980 to 31 December 2013 using MEDLINE, Cochrane Central Register of Controlled Trials, Science Citation Index Expanded, Social Sciences Citation Index, African Index Medicus, and CAB Abstracts. We stratified studies into three levels of insecticide resistance, and ITNs were compared with untreated bed nets (UTNs) using the risk difference (RD). Heterogeneity was explored visually and statistically. Included were 36 laboratory and 24 field studies, reported in 25 records. Studies tested and reported resistance inconsistently. Based on the meta-analytic results, the difference in mosquito mortality risk for ITNs compared to UTNs was lower in higher resistance categories. However, mortality risk was significantly higher for ITNs compared to UTNs regardless of resistance. For cone tests: low resistance, risk difference (RD) 0.86 (95% CI 0.72 to 1.01); moderate resistance, RD 0.71 (95% CI 0.53 to 0.88); high resistance, RD 0.56 (95% CI 0.17 to 0.95). For tunnel tests: low resistance, RD 0.74 (95% CI 0.61 to 0.87); moderate resistance, RD 0.50 (95% CI 0.40 to 0.60); high resistance, RD 0.39 (95% CI 0.24 to 0.54). For hut studies: low resistance, RD 0.56 (95% CI 0.43 to 0.68); moderate resistance, RD 0.39 (95% CI 0.16 to 0.61); high resistance, RD 0.35 (95% CI 0.27 to 0.43). However, with the exception of the moderate resistance category for tunnel tests, there was extremely high heterogeneity across studies in each resistance category (chi-squared test, p<0.00001, I2 varied from 95% to 100%). Conclusions This meta-analysis found that ITNs are more effective than UTNs regardless of resistance. There appears to be a relationship between resistance and the RD for mosquito mortality in laboratory and field studies. However, the substantive heterogeneity in the studies' results and design may mask the true relationship between resistance and the RD, and the results need to be interpreted with caution. Our analysis suggests the potential for cumulative meta-analysis in entomological trials, but further field research in this area will require specialists in the field to work together to improve the quality of trials, and to standardise designs, assessment, and reporting of both resistance and entomological outcomes

Malaria in Africa: Vector Species' Niche Models and Relative Risk Maps

A central theoretical goal of epidemiology is the construction of spatial models of disease prevalence and risk, including maps for the potential spread of infectious disease. We provide three continent-wide maps representing the relative risk of malaria in Africa based on ecological niche models of vector species and risk analysis at a spatial resolution of 1 arc-minute (9 185 275 cells of approximately 4 sq km). Using a maximum entropy method we construct niche models for 10 malaria vector species based on species occurrence records since 1980, 19 climatic variables, altitude, and land cover data (in 14 classes). For seven vectors (Anopheles coustani, A. funestus, A. melas, A. merus, A. moucheti, A. nili, and A. paludis) these are the first published niche models. We predict that Central Africa has poor habitat for both A. arabiensis and A. gambiae, and that A. quadriannulatus and A. arabiensis have restricted habitats in Southern Africa as claimed by field experts in criticism of previous models. The results of the niche models are incorporated into three relative risk models which assume different ecological interactions between vector species. The “additive” model assumes no interaction; the “minimax” model assumes maximum relative risk due to any vector in a cell; and the “competitive exclusion” model assumes the relative risk that arises from the most suitable vector for a cell. All models include variable anthrophilicity of vectors and spatial variation in human population density. Relative risk maps are produced from these models. All models predict that human population density is the critical factor determining malaria risk. Our method of constructing relative risk maps is equally general. We discuss the limits of the relative risk maps reported here, and the additional data that are required for their improvement. The protocol developed here can be used for any other vector-borne disease