Showing papers in "Malaria Journal in 2006"

Human malarial disease: a consequence of inflammatory cytokine release

Abstract: Malaria causes an acute systemic human disease that bears many similarities, both clinically and mechanistically, to those caused by bacteria, rickettsia, and viruses. Over the past few decades, a literature has emerged that argues for most of the pathology seen in all of these infectious diseases being explained by activation of the inflammatory system, with the balance between the pro and anti-inflammatory cytokines being tipped towards the onset of systemic inflammation. Although not often expressed in energy terms, there is, when reduced to biochemical essentials, wide agreement that infection with falciparum malaria is often fatal because mitochondria are unable to generate enough ATP to maintain normal cellular function. Most, however, would contend that this largely occurs because sequestered parasitized red cells prevent sufficient oxygen getting to where it is needed. This review considers the evidence that an equally or more important way ATP deficency arises in malaria, as well as these other infectious diseases, is an inability of mitochondria, through the effects of inflammatory cytokines on their function, to utilise available oxygen. This activity of these cytokines, plus their capacity to control the pathways through which oxygen supply to mitochondria are restricted (particularly through directing sequestration and driving anaemia), combine to make falciparum malaria primarily an inflammatory cytokine-driven disease.

A steep decline of malaria morbidity and mortality trends in Eritrea between 2000 and 2004: the effect of combination of control methods

Abstract: Malaria is a huge public health problem in Africa that is responsible for more than one million deaths annually. In line with the Roll Back Malaria initiative and the Abuja Declaration, Eritrea and other African countries have intensified their fight against malaria. This study examines the impact of Eritrea's Roll Back Malaria Programme: 2000–2004 and the effects and possible interactions between the public health interventions in use. This study employed cross-sectional survey to collect data from households, community and health facilities on coverage and usage of Insecticide-Treated Nets (ITNs), Indoor Residual Spraying (IRS), larvicidal activities and malaria case management. Comparative data was obtained from a similar survey carried out in 2001. Data from the Health Management Information System (HMIS) and reports of the annual assessments by the National Malaria Control Programme was used to assess impact. Time series model (ARIMA) was used to assess association. In the period 2000–2004, approximately 874,000 ITNs were distributed and 13,109 health workers and community health agents were trained on malaria case management. In 2004, approximately 81% households owned at least one net, of which 73% were ITNs and 58.6% of children 0–5 years slept under a net. The proportion of malaria cases managed by community health agents rose from 50% in 1999 to 78% in 2004. IRS coverage increased with the combined amount of DDT and Malathion used rising from 6,444 kg, in 2000 to 43,491 kg, in 2004, increasing the population protected from 117,017 to 259,420. Drug resistance necessitated regimen change to chloroquine plus sulfadoxine-pyrimethamine. During the period, there was a steep decline in malaria morbidity and case fatality by 84% and 40% respectively. Malaria morbidity was strongly correlated to the numbers of ITNs distributed (β = -0.125, p < 0.005) and the amount (kg) of DDT and Malathion used for IRS (β = -2.352, p < 0.05). The correlation between malaria case fatality and ITNs, IRS, population protected and annual rainfall was not statistically significant. Eritrea has within 5 years attained key Roll Back Malaria targets. ITNs and IRS contributed most to reducing malaria morbidity.

Real-time quantitative PCR with SYBR Green I detection for estimating copy numbers of nine drug resistance candidate genes in Plasmodium falciparum.

Abstract: Evaluating copy numbers of given genes in Plasmodium falciparum parasites is of major importance for laboratory-based studies or epidemiological surveys. For instance, pfmdr1 gene amplification has been associated with resistance to quinine derivatives and several genes involved in anti-oxidant defence may play an important role in resistance to antimalarial drugs, although their potential involvement has been overlooked. The ΔΔCt method of relative quantification using real-time quantitative PCR with SYBR Green I detection was adapted and optimized to estimate copy numbers of three genes previously indicated as putative candidates of resistance to quinolines and artemisinin derivatives: pfmdr1, pfatp6 (SERCA) and pftctp, and in six further genes involved in oxidative stress responses. Using carefully designed specific RT-qPCR oligonucleotides, the methods were optimized for each gene and validated by the accurate measure of previously known number of copies of the pfmdr1 gene in the laboratory reference strains P. falciparum 3D7 and Dd2. Subsequently, Standard Operating Procedures (SOPs) were developed to the remaining genes under study and successfully applied to DNA obtained from dried filter blood spots of field isolates of P. falciparum collected in Sao Tome & Principe, West Africa. The SOPs reported here may be used as a high throughput tool to investigate the role of these drug resistance gene candidates in laboratory studies or large scale epidemiological surveys.

The co-distribution of Plasmodium falciparum and hookworm among African schoolchildren.

Abstract: Surprisingly little is known about the geographical overlap between malaria and other tropical diseases, including helminth infections. This is despite the potential public health importance of co-infection and synergistic opportunities for control. Statistical models are presented that predict the large-scale distribution of hookworm in sub-Saharan Africa (SSA), based on the relationship between prevalence of infection among schoolchildren and remotely sensed environmental variables. Using a climate-based spatial model of the transmission potential for Plasmodium falciparum malaria, adjusted for urbanization, the spatial congruence of populations at coincident risk of infection is determined. The model of hookworm indicates that the infection is widespread throughout Africa and that, of the 179.3 million school-aged children who live on the continent, 50.0 (95% CI: 48.9–51.1) million (27.9% of total population) are infected with hookworm and 45.1 (95% CI: 43.9–46) million are estimated to be at risk of coincident infection. Malaria and hookworm infection are widespread throughout SSA and over a quarter of school-aged children in sub-Saharan Africa appear to be at risk of coincident infection and thus at enhanced risk of clinical disease. The results suggest that the control of parasitic helminths and of malaria in school children could be viewed as essential co-contributors to promoting the health of schoolchildren.

Malaria hotspot areas in a highland Kenya site are consistent in epidemic and non-epidemic years and are associated with ecological factors

Abstract: Background Malaria epidemics in highland areas of East Africa have caused considerable morbidity and mortality in the past two decades. Knowledge of "hotspot" areas of high malaria incidence would allow for focused preventive interventions in resource-poor areas, particularly if the hotspot areas can be discerned during non-epidemic periods and predicted by ecological factors.

Comparison of PCR and microscopy for the detection of asymptomatic malaria in a Plasmodium falciparum/vivax endemic area in Thailand

Abstract: The main objective of this study was to compare the performance of nested PCR with expert microscopy as a means of detecting Plasmodium parasites during active malaria surveillance in western Thailand. The study was performed from May 2000 to April 2002 in the village of Kong Mong Tha, located in western Thailand. Plasmodium vivax (PV) and Plasmodium falciparum (PF) are the predominant parasite species in this village, followed by Plasmodium malariae (PM) and Plasmodium ovale (PO). Each month, fingerprick blood samples were taken from each participating individual and used to prepare thick and thin blood films and for PCR analysis. PCR was sensitive (96%) and specific (98%) for malaria at parasite densities ≥ 500/μl; however, only 18% (47/269) of P. falciparum- and 5% (20/390) of P. vivax-positive films had parasite densities this high. Performance of PCR decreased markedly at parasite densities <500/μl, with sensitivity of only 20% for P. falciparum and 24% for P. vivax at densities <100 parasites/μl. Although PCR performance appeared poor when compared to microscopy, data indicated that the discrepancy between the two methods resulted from poor performance of microscopy at low parasite densities rather than poor performance of PCR. These data are not unusual when the diagnostic method being evaluated is more sensitive than the reference method. PCR appears to be a useful method for detecting Plasmodium parasites during active malaria surveillance in Thailand.

Landscape determinants and remote sensing of anopheline mosquito larval habitats in the western Kenya highlands

Abstract: In the past two decades the east African highlands have experienced several major malaria epidemics. Currently there is a renewed interest in exploring the possibility of anopheline larval control through environmental management or larvicide as an additional means of reducing malaria transmission in Africa. This study examined the landscape determinants of anopheline mosquito larval habitats and usefulness of remote sensing in identifying these habitats in western Kenya highlands. Panchromatic aerial photos, Ikonos and Landsat Thematic Mapper 7 satellite images were acquired for a study area in Kakamega, western Kenya. Supervised classification of land-use and land-cover and visual identification of aquatic habitats were conducted. Ground survey of all aquatic habitats was conducted in the dry and rainy seasons in 2003. All habitats positive for anopheline larvae were identified. The retrieved data from the remote sensors were compared to the ground results on aquatic habitats and land-use. The probability of finding aquatic habitats and habitats with Anopheles larvae were modelled based on the digital elevation model and land-use types. The misclassification rate of land-cover types was 10.8% based on Ikonos imagery, 22.6% for panchromatic aerial photos and 39.2% for Landsat TM 7 imagery. The Ikonos image identified 40.6% of aquatic habitats, aerial photos identified 10.6%, and Landsate TM 7 image identified 0%. Computer models based on topographic features and land-cover information obtained from the Ikonos image yielded a misclassification rate of 20.3–22.7% for aquatic habitats, and 18.1–25.1% for anopheline-positive larval habitats. One-metre spatial resolution Ikonos images combined with computer modelling based on topographic land-cover features are useful tools for identification of anopheline larval habitats, and they can be used to assist to malaria vector control in western Kenya highlands.

Topography and malaria transmission heterogeneity in western Kenya highlands: prospects for focal vector control

Abstract: Recent resurgence of malaria in the highlands of Western Kenya has called for a more comprehensive understanding of the previously neglected complex highland vector ecology. Besides other drivers of malaria epidemiology, topography is likely to have a major effect on spatial vector and parasite distribution. The aim of this study was to determine the effects of topography on malaria spatial vector distribution and parasite prevalence. Indoor resting adult malaria vectors and blood parasites were collected in three villages along a 4 km transect originating from the valley bottom and ending at the hilltop for 13 months. Members of the Anopheles gambiae complex were identified by PCR. Blood parasites were collected from children 6–13 years old and densities categorized by site of home location and age of the children. Ninety eight percent (98%) of An. gambiae s.s. and (99%) Anopheles funestus were collected in houses located at the edge of the valley bottom, whereas 1% of An. gambiae s.s. were collected at mid hill and at the hilltop respectively. No An. funestus were collected at the hilltop. Malaria prevalence was 68% at the valley bottom, 40.2% at mid hill and 26.7% at the hilltop. Children aged six years and living at the edge of the valley bottom had an annual geometric mean number of 66.1 trophozoites for every 200 white blood cells, while those living at mid-hill had a mean of 84.8, and those living at hilltop had 199.5 trophozoites. Malaria transmission in this area is mainly confined to the valley bottom. Effective vector control could be targeted at the foci. However, the few vectors observed at mid-hill maintained a relatively high prevalence rate. The higher variability in blood parasite densities and their low correlation with age in children living at the hilltop suggests a lower stability of transmission than at the mid-hill and valley bottom.

Spatial and temporal variation in malaria transmission in a low endemicity area in northern Tanzania.

Abstract: Background: Spatial and longitudinal monitoring of transmission intensity will allow better targeting of malaria interventions. In this study, data on meteorological, demographic, entomological and parasitological data over the course of a year was collected to describe malaria epidemiology in a single village of low transmission intensity. Methods: Entomological monitoring of malaria vectors was performed by weekly light trap catches in 10 houses. Each house in the village of Msitu wa Tembo, Lower Moshi, was mapped and censused. Malaria cases identified through passive case detection at the local health centre were mapped by residence using GIS software and the incidence of cases by season and distance to the main breeding site was calculated. Results: The principle vector was Anopheles arabiensis and peak mosquito numbers followed peaks in recent rainfall. The entomological inoculation rate estimated was 3.4 (95% CI 0.7–9.9) infectious bites per person per year. The majority of malaria cases (85/130) occurred during the rainy season (χ2 = 62,3, p < 0.001). Living further away from the river (OR 0.96, CI 0.92–0.998, p = 0.04 every 50 m) and use of anti-insect window screens (OR 0.65, CI 0.44–0.94, p = 0.023) were independent protective factors for the risk of malaria infection. Children aged 1–5 years and 5–15 years were at greater risk of clinical episodes (OR 2.36, CI 1.41–3.97, p = 0.001 and OR 3.68, CI 2.42–5.61, p < 0.001 respectively). Conclusion: These data show that local malaria transmission is restricted to the rainy season and strongly associated with proximity to the river. Transmission reducing interventions should, therefore, be timed before the rain-associated increase in mosquito numbers and target households located near the river.

Detection of the East and West African kdr mutation in Anopheles gambiae and Anopheles arabiensis from Uganda using a new assay based on FRET/Melt Curve analysis

Abstract: Appropriate monitoring of vector resistance to insecticides is an integral component of planning and evaluation of insecticide use in malaria control programmes. The malaria vectors Anopheles gambiae s.s. and Anopheles arabiensis have developed resistance to pyrethroid insecticides as a result of a mechanism conferring reduced nervous system sensitivity, better known as knockdown resistance (kdr). In An. gambiae s.s. and An. arabiensis, two different substitutions in the para-type sodium channel, a L1014F substitution common in West Africa and a L1014S replacement found in Kenya, are linked with kdr. Two different allele-specific polymerase chain reactions (AS-PCR) are needed to detect these known kdr mutations. However, these AS-PCR assays rely on a single nucleotide polymorphism mismatch, which can result in unreliable results. Here, a new assay for the detection of knockdown resistance in An. gambiae s.s. and An. arabiensis based on Fluorescence Resonance Energy Transfer/Melt Curve analysis (FRET/MCA) is presented and compared with the existing assays. The new FRET/MCA method has the important advantage of detecting both kdr alleles in one assay. Moreover, results show that the FRET/MCA is more reliable and more sensitive than the existing AS-PCR assays and is able to detect new genotypes. By using this technique, the presence of the East African kdr mutation (L1014S) is shown for the first time in An. arabiensis specimens from Uganda. In addition, a new kdr genotype is reported in An. gambiae s.s. from Uganda, where four An. gambiae s.s. mosquitoes possess both, the West (L1014F) and East (L1014S) African kdr allele, simultaneously. The presence of both kdr mutations in the same geographical region shows the necessity of a reliable assay that enables to detect both mutations in one single assay. Hence, this new assay based on FRET/MCA will improve the screening of the kdr frequencies in An. gambiae s.s. and An. arabiensis.

Trapping of the malaria vector Anopheles gambiae with odour-baited MM-X traps in semi-field conditions in western Kenya

Abstract: Background: The successful development of odour-baited trapping systems for mosquitoes depends on the identification of behaviourally active semiochemicals, besides the design and operating principles of such devices. A large variety of 'attractants' has been identified in laboratory investigations, yet few of these increase trap catches in the field. A contained system, intermediate between the laboratory and open field, is presented and previous reports that human foot odour induces behavioural responses of Anopheles gambiae confirmed. Methods: The response of 3–5 day old female An. gambiae towards odour-baited counterflow geometry traps (MM-X model; American Biophysics Corp., RI) was studied in semi-field (screen house) conditions in western Kenya. Traps were baited with human foot odour (collected on socks), carbon dioxide (CO2, 500 ml min-1), ammonia (NH3), 1-octen-3-ol, or various combinations thereof. Trap catches were log (x+1) transformed and subjected to Latin square analysis of variance procedures. Results: Apart from 1-octen-3-ol, all odour baits caused significant (P < 0.05) increases in trap catches over non-baited traps. Foot odour remained behaviourally active for at least 8 days after collection on nylon or cotton sock fabric. A synergistic response (P < 0.001) was observed towards the combination of foot odour and CO2, which increased catches of these odours alone by 3.8 and 2.7 times, respectively. Conclusion: These results are the first to report behavioural responses of an African malaria vector to human foot odour outside the laboratory, and further investigation of fractions and/or individual chemical components of this odour complex are called for. Semi-field systems offer the prospect of high-throughput screening of candidate kairomones, which may expedite the development of efficient trap-bait systems for this and other African mosquito species.

Rethinking the economic costs of malaria at the household level: evidence from applying a new analytical framework in rural Kenya.

Abstract: Malaria imposes significant costs on households and the poor are disproportionately affected. However, cost data are often from quantitative surveys with a fixed recall period. They do not capture costs that unfold slowly over time, or seasonal variations. Few studies investigate the different pathways through which malaria contributes towards poverty. In this paper, a framework indicating the complex links between malaria, poverty and vulnerability at the household level is developed and applied using data from rural Kenya. Cross-sectional surveys in a wet and dry season provide data on treatment-seeking, cost-burdens and coping strategies (n = 294 and n = 285 households respectively). 15 case study households purposively selected from the survey and followed for one year provide in-depth qualitative information on the links between malaria, vulnerability and poverty. Mean direct cost burdens were 7.1% and 5.9% of total household expenditure in the wet and dry seasons respectively. Case study data revealed no clear relationship between cost burdens and vulnerability status at the end of the year. Most important was household vulnerability status at the outset. Households reporting major malaria episodes and other shocks prior to the study descended further into poverty over the year. Wealthier households were better able to cope. The impacts of malaria on household economic status unfold slowly over time. Coping strategies adopted can have negative implications, influencing household ability to withstand malaria and other contingencies in future. To protect the poor and vulnerable, malaria control policies need to be integrated into development and poverty reduction programmes.

Radiation-induced sterility for pupal and adult stages of the malaria mosquito Anopheles arabiensis

Abstract: In the context of the Sterile Insect Technique (SIT), radiation-induced sterility in the malaria mosquito Anopheles arabiensis Patton (Diptera: Culicidae) was studied. Male mosquitoes were exposed to gamma rays in the pupal or adult stage and dose-sterility curves were determined. Pupae were irradiated shortly before emergence (at 22–26 hrs of age), and adults <24 hrs post emergence. Doses tested ranged between 0 and 100 Gy. The effects of irradiation on adult emergence, male survival, induced sterility and insemination capability were evaluated. Emergence and insemination data were analysed using independent t-tests against the control. Correlation analyses were performed for insemination rate and dose and insemination and fecundity. Male survival was analysed using Kaplan-Meier survival analyses. Finally, the calculated residual fertility values were inverse-normal transformed and linear regression analyses performed. Irradiation of pupae, for all doses tested, had no effect on adult emergence. Survival curves of males irradiated as pupae or adults were similar or even slightly higher than non-irradiated males. Overall, adults appeared to be slightly more susceptible to irradiation, although no significant differences for individual doses were observed. In the pupal stage, a significant negative correlation was found between insemination and dose, but the correlation-coefficient was associated with less than 25% of the total variation. A review of the literature indicated that An. arabiensis is more radiation resistant than other anopheline mosquitoes. The optimal dose for male insects to be released in an SIT programme depends on their level of sterility and competitiveness. The use of semi-sterilizing doses to produce more competitive insects is discussed. The most convenient developmental stage for mosquito irradiation on a mass-scale are pupae, but pupal irradiation resulted in a lower insemination rate at the highest dose compared to adult irradiation. On the basis of this study, a suitable dose range that includes semi-sterilizing doses is identified to initiate competitiveness experiments for males irradiated at both developmental stages.

Role of traditional healers in the management of severe malaria among children below five years of age: the case of Kilosa and Handeni Districts, Tanzania.

Abstract: Background: The current malaria control strategy of WHO centres on early diagnosis and prompt treatment using effective drugs. Children with severe malaria are often brought late to health facilities and traditional health practitioners are said to be the main cause of treatment delay. In the context of the Rectal Artesunate Project in Tanzania, the role of traditional healers in the management of severe malaria in children was studied. Methodology: A community cross-sectional study was conducted in Kilosa and Handeni Districts, involving four villages selected on the basis of existing statistics on the number of traditional health practitioners involved in the management of severe malaria. A total of 41 traditional health practitioners were selected using the snowballing technique, whereby in-depth interviews were used to collect information. Eight Focus Group Discussions (FGDs) involving traditional health practitioners, caregivers and community leaders were carried out in each district. Results: Home management of fever involving sponging or washing with warm water at the household level, was widely practiced by caregivers. One important finding was that traditional health practitioners and mothers were not linking the local illness termed degedege, a prominent feature in severe malaria, to biomedically-defined malaria. The majority of mothers (75%) considered degedege to be caused by evil spirits. The healing process was therefore organized in stages and failure to abide to the procedure could lead to relapse of degedege, which was believed to be caused by evil spirits. Treatment seeking was, therefore, a complex process and mothers would consult traditional health practitioners and modern health care providers, back and forth. Referrals to health facilities increased during the Rectal Artesunate Project, whereby project staff facilitated the process after traditional medical care with the provision of suppositories. This finding is challenging the common view that traditional healers are an important factor of delay for malaria treatment, they actually play a pivotal role by giving "bio-medically accepted first aid" which leads to reduction in body temperature hence increasing chances of survival for the child. Increasing the collaboration between traditional healers and modern health care providers was shown to improve the management of severe malaria in the studied areas. Interpretation and conclusion: Traditional health care is not necessarily a significant impediment or a delaying factor in the treatment of severe malaria. There is a need to foster training on the management of severe cases, periodically involving both traditional health practitioners and health workers to identify modalities of better collaboration.

Thick blood film examination for Plasmodium falciparum malaria has reduced sensitivity and underestimates parasite density.

Abstract: Background: Thick blood films are routinely used to diagnose Plasmodium falciparum malaria. Here, they were used to diagnose volunteers exposed to experimental malaria challenge. Methods: The frequency with which blood films were positive at given parasite densities measured by PCR were analysed. The poisson distribution was used to calculate the theoretical likelihood of diagnosis. Further in vitro studies used serial dilutions to prepare thick films from malaria cultures at known parasitaemia. Results: Even in expert hands, thick blood films were considerably less sensitive than might have been expected from the parasite numbers measured by quantitative PCR. In vitro work showed that thick films prepared from malaria cultures at known parasitaemia consistently underestimated parasite densities. Conclusion: It appears large numbers of parasites are lost during staining. This limits their sensitivity, and leads to erroneous estimates of parasite density.

Use of insecticide-treated clothes for personal protection against malaria: a community trial.

Abstract: The study sought to determine the effect of using insecticide-treated clothes (ITCs) on personal protection against malaria infection. The specific objectives were to determine the effect of using ITCs on the rate of infection with malaria parasites and the effect on indoor mosquito density. This study was done in Dadaab refugee camps, North Eastern Province Kenya between April and August 2002, and involved a total of 198 participants, all refugees of Somali origin. The participants were selected through multi-stage cluster sampling. Half of the participants (treatment group) had their personal clothes worn on a daily basis (Diras, Saris, Jalbaab s, Ma'awis and shirts) and their bedding (sheets and blankets) treated with insecticide (permethrin). The other half (comparison group) had their clothes treated with placebo (plain water). Indoor mosquito density was determined from twelve households belonging to the participants; six in the treatment block and six in the comparison block. During pre-test and post-test, laboratory analysis of blood samples was done, indoor mosquito density determined and questionnaires administered. Using STATA statistical package, tests for significant difference between the two groups were conducted. Use of ITCs reduced both malaria infection rates and indoor mosquito density significantly. The odds of malaria infection in the intervention group were reduced by about 70 percent. The idea of using ITCs for malaria infection control was easily accepted among the refugees and they considered it beneficial. No side effects related to use of the ITCs were observed from the participants. The use of ITCs reduces malaria infection rate and has potential as an appropriate method of malaria control. It is recommended, therefore, that this strategy be considered for use among poor communities like slum dwellers and other underprivileged communities, such as street children and refugees, especially during an influx to malaria-prone regions. Further research on cost-effectiveness and sustainability of this strategy is worthwhile.

Methylene blue for malaria in Africa: results from a dose-finding study in combination with chloroquine

Abstract: The development of safe, effective and affordable drug combinations against malaria in Africa is a public health priority. Methylene blue (MB) has a similar mode of action as chloroquine (CQ) and has moreover been shown to selectively inhibit the Plasmodium falciparum glutathione reductase. In 2004, an uncontrolled dose-finding study on the combination MB-CQ was performed in 435 young children with uncomplicated falciparum malaria in Burkina Faso (CQ monotherapy had a > 50% clinical failure rate in this area in 2003). Three serious adverse events (SAE) occurred of which one was probably attributable to the study medication. In the per protocol safety analysis, there were no dose specific effects. The overall clinical and parasitological failure rates by day 14 were 10% [95% CI (7.5%, 14.0%)] and 24% [95% CI (19.4%, 28.3%)], respectively. MB appears to have efficacy against malaria, but the combination of CQ-MB is clearly not effective in the treatment of malaria in Africa.

The contribution of microscopy to targeting antimalarial treatment in a low transmission area of Tanzania.

Abstract: Background There is a need for improved targeting of antimalarial treatment if artemisinin combination therapy is to be successfully introduced in Africa. This study aimed to explore why malaria slides are requested and how their results guide treatment decisions in an area of low transmission of P. falciparum.

Screening of pesticide residues in soil and water samples from agricultural settings

Abstract: The role of agricultural practices in the selection of insecticide resistance in malaria vectors has so far been hypothesized without clear evidence. Many mosquito species, Anopheles gambiae in particular, lay their eggs in breeding sites located around agricultural settings. There is a probability that, as a result of farming activities, insecticide residues may be found in soil and water, where they exercise a selection pressure on the larval stage of various populations of mosquitoes. To confirm this hypothesis, a study was conducted in the Republic of Benin to assess the environmental hazards which can be generated from massive use of pesticides in agricultural settings. Lacking an HPLC machine for direct quantification of insecticide residues in samples, this investigation was performed using indirect bioassays focussed on the study of factors inhibiting the normal growth of mosquito larvae in breeding sites. The speed of development was monitored as well as the yield of rearing An. gambiae larvae in breeding sites reconstituted with water and soil samples collected in agricultural areas known to be under pesticide pressure. Two strains of An. gambiae were used in this indirect bioassay: the pyrethroid-susceptible Kisumu strain and the resistant Ladji strain. The key approach in this methodology is based on comparison of the growth of larvae in test and in control breeding sites, the test samples having been collected from two vegetable farms. Results obtained clearly show the presence of inhibiting factors on test samples. A normal growth of larvae was observed in control samples. In breeding sites simulated by using a few grams of soil samples from the two vegetable farms under constant insecticide treatments (test samples), a poor hatching rate of Anopheles eggs coupled with a retarded growth of larvae and a low yield of adult mosquitoes from hatched eggs, was noticed. Toxic factors inhibiting the hatching of anopheles eggs and the growth of larvae are probably pesticide residues from agricultural practices. Samples used during this indirect assay have been stored in the laboratory and will be analysed with HPLC techniques to confirm hypothesis of this study and to identify the various end products found in soil and water samples from agricultural settings under pesticide pressure.

Housing conditions and Plasmodium falciparum infection: protective effect of iron-sheet roofed houses

Abstract: Identification and better understanding of potential risk factors for malaria are important for targeted and cost-effective health interventions. Housing conditions have been suggested as one of the potential risk factors. This study aims to further investigate this risk factor, and is focused on the effect of the type of roof on Plasmodium falciparum infection among children below five years in the North West of Burkina Faso. In a cross-sectional study design, 661 children aged six to 60 months were randomly selected from three rural and one semi-urban site at the end of the rainy season (November 2003). The children were screened for fever and tested for Plasmodium falciparum infection. In addition, data on bed net use and house characteristics was collected from the household were each child lived. Using adjusted odds ratios, children living in house roofed with iron-sheet were compared with those in house with mud or grass roof. Overall P. falciparum infection prevalence was 22.8 % with a significant variation between (Chi-square, p < 0.0001). The prevalence in Cisse (33.3 %) and Goni (30.6 %) were twice times more than in Nouna (15.2 %) and Kodougou (13.2 %). After adjusting for age, sex, use of bed net and housing conditions, children living in houses with mud roofs had significantly higher risk of getting P. falciparum infection compared to those living in iron-sheet roofed houses (Odds Ratio 2.6; 95% Confidence Interval, 1.4–4.7). These results suggest that house characteristics should be taken into consideration when designing health intervention against P. falciparum infection and particular attention should be paid to children living in houses with mud roofs.

Occurrence of the leucine-to-phenylalanine knockdown resistance (kdr) mutation in Anopheles arabiensis populations in Tanzania, detected by a simplified high-throughput SSOP-ELISA method

Abstract: Molecular markers of insecticide resistance can provide sensitive indicators of resistance development in malaria vector populations. Monitoring of insecticide resistance in vector populations is an important component of current malaria control programmes. Knockdown resistance (kdr) confers resistance to the pyrethroid class of insecticides with cross-resistance to DDT through single nucleotide polymorphisms (SNPs) in the voltage-gated sodium channel gene. To enable detection of kdr mutations at low frequency a method was developed that uses polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA)-based technology, allowing rapid, reliable and cost-effective testing of large numbers of individual mosquitoes. This was used to assay mosquitoes from sites in lower Moshi, Tanzania. Sequence-specific oligonucleotide probes (SSOP) were used for simultaneous detection of both East and West African kdr mutations with high specificity and sensitivity. Application of the SSOP-ELISA method to 1,620 field-collected Anopheles arabiensis from Tanzania identified the West African leucine-phenylalanine kdr mutation in two heterozygous individuals, indicating the potential for resistance development that requires close monitoring. The presence of the West African kdr mutation at low frequency in this East African population of An. arabiensis has implications for the spread of the kdr gene across the African continent.

PCR detection of Plasmodium falciparum in human urine and saliva samples.

Abstract: Current detection or screening for malaria infection necessitates drawing blood by fingerprick or venipuncture, which poses risks and limitations for repeated measurement. This study presents PCR detection of Plasmodium falciparum in human urine and saliva samples, and illustrates this potential application in genotyping malaria infections. Urine and saliva were obtained from 47 thick film positive and 4 negative individuals one day after collection of blood slides and filter paper blood spots. P. falciparum DNA was extracted from blood, urine and saliva, in separate groups, using the Chelex method or Qiagen DNEasy® kit (urine and saliva only). Blood, urine and saliva extracts were subjected to PCR in separate batches. Amplicons from the various sample types were examined for MSP2 polymorphisms and restriction fragment patterns on DHFR amino acid codon 59. Malaria infections exhibited primarily low-grade parasite densities, with a geometric mean of 775 asexual parasites/μl. Regularly matching polymorphic MSP2 genotypes were found between the corresponding urine, saliva and peripheral blood amplicons of each individual, with different inter-individual polymorphic genotypes. Amplicon yields were significantly dependent on DNA extraction method, parasite density and primer set (p < 0.001). A Qiagen® kit extraction had more than 2× higher amplicon yield than the Chelex method, for both urine and saliva. Amplicon yields were 1.6 fold higher from saliva than urine. For each unit increase in log parasite density, the probability of amplicon enhanced 1.8 fold. Highest amplicon yields were obtained from the primer set with the shortest PCR product. P. falciparum infection is detectable by PCR on human urine and saliva samples. Subject to further refinement of extraction technique and amplicon yields, large-scale malaria parasite screening and epidemiological surveys could be possible without the need to collect blood and use of needles or sharps.

Detection and clinical manifestation of placental malaria in southern Ghana

Abstract: Background: Plasmodium falciparum can be detected by microscopy, histidine-rich-protein-2 (HRP2) capture test or PCR but the respective clinical relevance of the thereby diagnosed infections in pregnant women is not well established. Methods: In a cross-sectional, year-round study among 839 delivering women in Agogo, Ghana, P. falciparum was screened for in both, peripheral and placental blood samples, and associations with maternal anaemia, low birth weight (LBW) and preterm delivery (PD) were analysed. Results: In peripheral blood, P. falciparum was observed in 19%, 34%, and 53% by microscopy, HRP2 test, and PCR, respectively. For placental samples, these figures were 35%, 41%, and 59%. Irrespective of diagnostic tool, P. falciparum infection increased the risk of anaemia. Positive peripheral blood results of microscopy and PCR were not associated with LBW or PD. In contrast, the HRP2 test performed well in identifying women at increased risk of poor pregnancy outcome, particularly in case of a negative peripheral blood film. Adjusting for age, parity, and antenatal visits, placental HRP2 was the only marker of infection associated with LBW (adjusted odds ratio (aOR), 1.5 (95%CI, 1.0–2.2)) and, at borderline statistical significance, PD (aOR, 1.4 (1.0–2.1)) in addition to anaemia (aOR, 2.3 (1.7–3.2)). Likewise, HRP2 in peripheral blood of seemingly aparasitaemic women was associated with PD (aOR, 1.7 (1.0–2.7)) and anaemia (aOR, 2.1 (1.4–3.2)). Conclusion: Peripheral blood film microscopy not only underestimates placental malaria. In this highly endemic setting, it also fails to identify malaria as a cause of foetal impairment. Sub-microscopic infections detected by a HRP2 test in seemingly aparasitaemic women increase the risks of anaemia and PD. These findings indicate that the burden of malaria in pregnancy may be even larger than thought and accentuate the need for effective anti-malarial interventions in pregnancy.

An in-depth study of patent medicine sellers' perspectives on malaria in a rural Nigerian community

Abstract: Malaria remains a major cause of mortality among under five children in Nigeria. Most of the early treatments for fever and malaria occur through self-medication with antimalarial drugs bought from medicine sellers. These have led to increasing calls for interventions to improve treatment obtained in these outlets. However, information about the current practices of these medicine sellers is needed before such interventions. This study aims to determine the medicine sellers' perspectives on malaria and the determinants that underlie their dispensing patterns of antimalarial drugs. The study was conducted in Ugwugo-Nike, a rural community in south-east Nigeria. It involved in-depth interviews with 13 patent medicine sellers. A majority of the medicine sellers were not trained health professionals and malaria is recognized as a major health problem by them. There is poor knowledge and poor dispensing behaviour in relation to childhood malaria episodes. Although referral of severe malaria is common, there are those who will not refer. Verbal advice is rarely given to the care-givers. More action research and interventions to improve prescription and referral practices and giving verbal advice to care-givers is recommended. Ways to integrate the drug sellers in the health system are also recommended.

Ecologists can enable communities to implement malaria vector control in Africa

Abstract: Integrated vector management (IVM) for malaria control requires ecological skills that are very scarce and rarely applied in Africa today. Partnerships between communities and academic ecologists can address this capacity deficit, modernize the evidence base for such approaches and enable future scale up. Community-based IVM programmes were initiated in two contrasting settings. On Rusinga Island, Western Kenya, community outreach to a marginalized rural community was achieved by University of Nairobi through a community-based organization. In Dar es Salaam, Tanzania, Ilala Municipality established an IVM programme at grassroots level, which was subsequently upgraded and expanded into a pilot scale Urban Malaria Control Programme with support from national academic institutes. Both programmes now access relevant expertise, funding and policy makers while the academic partners benefit from direct experience of community-based implementation and operational research opportunities. The communities now access up-to-date malaria-related knowledge and skills for translation into local action. Similarly, the academic partners have acquired better understanding of community needs and how to address them. Until sufficient evidence is provided, community-based IVM remains an operational research activity. Researchers can never directly support every community in Africa so community-based IVM strategies and tactics will need to be incorporated into undergraduate teaching programmes to generate sufficient numbers of practitioners for national scale programmes. Academic ecologists at African institutions are uniquely positioned to enable the application of practical environmental and entomological skills for malaria control by communities at grassroots level and should be supported to fulfil this neglected role.

Web-based climate information resources for malaria control in Africa.

Abstract: Malaria remains a major public health threat to more than 600 million Africans and its control is recognized as critical to achieving the Millennium Development Goals. The greatest burden of malaria in Africa occurs in the endemic regions where the disease pathogen is continuously present in the community. These regions are characterized by an environment that is conducive to interactions between the Anopheles mosquito, malaria parasites and human hosts, as well as housing of generally poor quality, which offers little protection from mosquito-human contact. Epidemic malaria tends to occur along the geographical margins of endemic regions, when the equilibrium between the human, parasite and mosquito vector populations is occasionally disturbed and a sharp but temporary increase in disease incidence results. When malaria control measures are inadequate, as is the case in much of sub-Saharan Africa, the disease distribution is closely linked with seasonal patterns of the climate and local environment. In the absence of good epidemiological data on malaria distribution in Africa, climate information has long been used to develop malaria risk maps that illustrate the boundaries of 'climatic suitability for endemic transmission.' The best known of these are produced by the Pan-African-based MARA Collaboration. This paper describes the development of additional malaria suitability maps which have been produced in an online, interactive format to enable temporal information (i.e., seasonality of climate conditions) to be queried and displayed along with spatial information. These maps and the seasonal information that they contain should be useful to the malaria control and health service communities for their planning and operational activities.

Genetic diversity of Plasmodium vivax in Kolkata, India.

Abstract: Plasmodium vivax malaria accounts for approximately 60% of malaria cases in Kolkata, India. There has been limited information on the genotypic polymorphism of P. vivax in this malaria endemic area. Three highly polymorphic and single copy genes were selected for a study of genetic diversity in Kolkata strains. Blood from 151 patients with P. vivax infection diagnosed in Kolkata between April 2003 and September 2004 was genotyped at three polymorphic loci: the P. vivax circumsporozoite protein (pvcs), the merozoite surface protein 1 (pvmsp 1) and the merozoite surface protein 3-alpha (pvmsp 3-alpha). Analysis of these three genetic markers revealed that P. vivax populations in Kolkata are highly diverse. A large number of distinguishable alleles were found from three genetic markers: 11 for pvcs, 35 for pvmsp 1 and 37 for pvmsp 3-alpha. These were, in general, randomly distributed amongst the isolates. Among the 151 isolates, 142 unique genotypes were detected the commonest genotype at a frequency of less than 2% (3/151). The overall rate of mixed genotype infections was 10.6%. These results indicate that the P. vivax parasite population is highly diverse in Kolkata, despite the low level of transmission. The genotyping protocols used in this study may be useful for differentiating re-infection from relapse and recrudescence in studies assessing of malarial drug efficacy in vivax malaria.

Treatment-seeking behaviour for malaria in children under five years of age: implication for home management in rural areas with high seasonal transmission in Sudan

Abstract: Background: Effective management of malaria in children under the age of 5 requires mothers to seek, obtain, and use medication appropriately. This is linked to timely decision, accessibility, correct use of the drugs and follow-up. The aim of the study is to identify the basis on which fever was recognized and classified and exploring factors involved in selection of different treatment options. Methods: Data was obtained by interviewing 96 mothers who had brought their febrile children to selected health facilities, conduction of 10 focus group discussions with mothers at village level as well as by observation. Results: A high score of mothers' knowledge and recognition of fever/malaria was recorded. Mothers usually start care at home and, within an average of three days, they shift to health workers if there was no response. The main health-seeking behaviour is to consult the nearest health facility or health personnel together with using traditional medicine or herbs. There are also health workers who visit patients at home. The majority of mothers with febrile children reported taking drugs before visiting a health facility. The choice between the available options determined by the availability of health facilities, user fees, satisfaction with services, difficulty to reach the facilities and believe in traditional medicine. Conclusion: Mothers usually go through different treatment option before consulting health facilities ending with obvious delay in seeking care. As early effective treatment is the main theme of the control programme, implementation of malaria home management strategy is urgently needed to improve the ongoing practice.

Who sleeps under bednets in Ghana? A doer/non-doer analysis of malaria prevention behaviours

Abstract: Malaria prevention programmes should be based in part on knowledge of why some individuals use bednets while others do not This paper identifies factors and characteristics of women that affect bednet use among their children less than five years of age in Ghana Data come from the baseline component of an evaluation of Freedom from Hunger's malaria curriculum A quasi-experimental design was used to select clients (n = 516) of Credit with Education (an integrated package of microfinance and health education) and non-clients (n = 535) Chi-squares, Fisher's Exact tests and logistic regression were used to compare the characteristics of mothers whose children use bednets (doers) with those whose children do not (non-doers) and to identify factors associated with bednet use among children less than five years of age The following factors were most closely associated with bednet use: region of residence; greater food security; and caregivers' beliefs about symptoms, causation and groups most vulnerable to malaria Most respondents knew mosquitoes caused malaria; however, 206% of doers and 123% of non-doers (p = 0228) thought overworking oneself caused malaria Ninety percent of doers and 770% of non-doers felt that sleeping under a net was protective against malaria (p = 0040) In addition, 165% of doers and 75% of non-doers (p = 0025) identified adult males as most vulnerable to malaria Greater knowledge about malaria does not always translate into improved bednet use Though culturally-based ideas about malaria may vary between communities, integrating them into traditional health education messages may enhance the effectiveness of public health efforts

Wealth, mother's education and physical access as determinants of retail sector net use in rural Kenya

Abstract: Insecticide-treated bed nets (ITN) provide real hope for the reduction of the malaria burden across Africa. Understanding factors that determine access to ITN is crucial to debates surrounding the optimal delivery systems. The influence of homestead wealth on use of nets purchased from the retail sector is well documented, however, the competing influence of mother's education and physical access to net providers is less well understood. Between December 2004 and January 2005, a random sample of 72 rural communities was selected across four Kenyan districts. Demographic, assets, education and net use data were collected at homestead, mother and child (aged < 5 years) levels. An assets-based wealth index was developed using principal components analysis, travel time to net sources was modelled using geographic information systems, and factors influencing the use of retail sector nets explored using a multivariable logistic regression model. Homestead heads and guardians of 3,755 children < 5 years of age were interviewed. Approximately 15% (562) of children slept under a net the night before the interview; 58% (327) of the nets used were purchased from the retail sector. Homestead wealth (adjusted OR = 10.17, 95% CI = 5.45–18.98), travel time to nearest market centres (adjusted OR = 0.51, 95% CI = 0.37–0.72) and mother's education (adjusted OR = 2.92, 95% CI = 1.93–4.41) were significantly associated with use of retail sector nets by children aged less than 5 years. Approaches to promoting access to nets through the retail sector disadvantage poor and remote communities where mothers are less well educated.