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BackgroundThe impact of insecticide treated nets (ITNs) on reducing malaria incidence is shown mainly through data collection from health facilities. Routine evaluation of long-term epidemiological and entomological dynamics is currently unavailable. In Kenya, new policies supporting the provision of free ITNs were implemented nationwide in June 2006. To evaluate the impacts of ITNs on malaria transmission, we conducted monthly surveys in three sentinel sites with different transmission intensities in western Kenya from 2002 to 2010.Methods and FindingsLongitudinal samplings of malaria parasite prevalence in asymptomatic school children and vector abundance in randomly selected houses were undertaken monthly from February 2002. ITN ownership and usage surveys were conducted annually from 2004 to 2010. Asymptomatic malaria parasite prevalence and vector abundances gradually decreased in all three sites from 2002 to 2006, and parasite prevalence reached its lowest level from late 2006 to early 2007. The abundance of the major malaria vectors, Anopheles funestus and An. gambiae, increased about 5–10 folds in all study sites after 2007. However, the resurgence of vectors was highly variable between sites and species. By 2010, asymptomatic parasite prevalence in Kombewa had resurged to levels recorded in 2004/2005, but the resurgence was smaller in magnitude in the other sites. Household ITN ownership was at 50–70% in 2009, but the functional and effective bed net coverage in the population was estimated at 40.3%, 49.4% and 28.2% in 2010 in Iguhu, Kombewa, and Marani, respectively.ConclusionThe resurgence in parasite prevalence and malaria vectors has been observed in two out of three sentinel sites in western Kenya despite a high ownership of ITNs. The likely factors contributing to malaria resurgence include reduced efficacy of ITNs, insecticide resistance in mosquitoes and lack of proper use of ITNs. These factors should be targeted to avoid further resurgence of malaria transmission.

Background: Interest in indoor residual spray (IRS) has been rekindled in recent years, as it is increasingly considered to be a key component of integrated malaria management. Regular spraying of each human dwelling becomes less and less practical as the control area increases. Where malaria transmission is concentrated around focal points, however, targeted IRS may pose a feasible alternative to mass spraying. Here, the impact of targeted IRS was assessed in the highlands of western Kenya. Methods: Indoor residual spray using lambda-cyhalothrin insecticide was carried out during the last week of April 2005 in 1,100 targeted houses, located in the valley bottom areas of Iguhu village, Kakamega district of western Kenya. Although the uphill areas are more densely populated, valleys are believed to be malaria transmission hotspots. The aim of the study was to measurably reduce the vector density and malaria transmission in uphill areas by focusing control on these hotspots. A cohort of 1,058 children from 1-5 yrs of age was randomly selected from a 4 km by 6 km study area for the baseline malaria prevalence survey after pre-clearing malaria infections during the third week of April 2005, and the prevalence of Plasmodium infections was tested bi-weekly. Seasonal changes in mosquito densities 12 months before the IRS and 12 months after the IRS was monitored quarterly based on 300 randomly selected houses. Monthly parasitological surveys were also carried out in the same area with 129-661 randomly selected school children of age 6-13 yrs. Results: The result of monthly parasitological surveys indicated that malaria prevalence in school children was reduced by 64.4% in the intervention valley area and by 46.3% in the intervention uphill area after 12 months of follow-ups in contrast to nonintervention areas (valley or uphill). The cohort study showed an average of 4.5% fewer new infections biweekly in the intervention valley compare to nonintervention valley and the relative reduction in incidence rate by week 14 was 65.4%. The relative reduction in incidence rate in intervention uphill by week 14 was 46.4%. Anopheles gambiae densities were reduced by 96.8% and 51.6% in the intervention valley and intervention uphill, respectively, and Anopheles funestus densities were reduced by 85.3% and 69.2% in the intervention valley and intervention uphill, respectively. Conclusion: Vector control had significant indirect impact on the densely populated uphill areas when IRS was targeted to the high-risk valleys. Additionally, the wide-reaching benefits of IRS in reducing vector prevalence and disease incidence was observed for at least six months following spraying, suggesting targeted IRS as an effective tool in malaria control.

Spatial and temporal variations in the distribution of anopheline larval habitats and land use and land cover (LULC) changes can influence malaria transmission intensity. This information is important for understanding the environmental determinants of malaria transmission heterogeneity, and it is critical to the study of the effects of environmental changes on malaria transmission. In this study, we investigated the spatial and temporal variations in the distribution of anopheline larval habitats and LULC changes in western Kenya highlands over a 4-year period. Anopheles gambiae complex larvae were mainly confined to valley bottoms during both the dry and wet seasons. Although An. gambiae larvae were located in man-made habitats where riparian forests and natural swamps had been cleared, Anopheles funestus larvae were mainly found in permanent habitats in pastures. The association between land cover type and occurrence of anopheline larvae was statistically significant. The distribution of anopheline positive habitats varied significantly between months, during the survey. In 2004, the mean density of An. gambiae was significantly higher during the month of May, whereas the density of An. funestus peaked significantly in February. Over the study period, major LULC changes occurred mostly in the valley bottoms. Overall, farmland increased by 3.9%, whereas both pastures and natural swamps decreased by 8.9% and 20.9%, respectively. The area under forest cover was decreased by 5.8%. Land-use changes in the study area are favorable to An. gambiae larval development, thereby risking a more widespread distribution of malaria vector habitats and potentially increasing malaria transmission in western Kenya highlands.

The number of Anopheles gambiae and Anopheles arabiensis females that used each of the 33 sampled breeding sites in west Kenya was estimated by microsatellite markers and related statistics to test the hypothesis that conspecific females share aquatic sites. Totally, 166 An. gambiae and 168 An. arabiensis larvae were identified and were genotyped. The mean number of larvae per breeding site was 8.3 for An. gambiae and 8.4 for An. arabiensis. The likelihood method estimated that, for An. gambiae, the mean number of females that would have laid eggs per breeding site was 5.2 and ranged from 2 to 9, and for An. arabiensis, the mean was 5.0 with a range of 2-10. The clustering method estimated that the mean number of females laying eggs per breeding site was 6.8 for An. gambiae. The results provide molecular evidence that females of one or both species share breeding sites. © 2007 Springer Science+Business Media, LLC.

We examined the survivorship of Anopheles gambiae s.l. Giles (Diptera: Culicidae) larvae and habitat productivity in three major habitat types in the western Kenya highlands. The age-specific distribution was determined for larvae and pupae, and survivorship curves were constructed. Larval-to-pupal survivorship was 6.8% in drainage ditches, 4.3% in cow hoofprints, and 1.8% in disused goldmines, respectively. High mortality rates were observed in all developmental stages. The estimated daily survival rate was highest in drainage ditches (0.74), followed by cow hoofprints (0.71), and it was lowest in disused goldmines (0.62). Productivity of emerging An. gambiae adults was generally low in these larval habitats (1.35, 1.55, and 1.84 mosquitoes per m2 per wk in drainage ditches, disused goldmines, and cow hoofprints, respectively). In total, seven families of larval mosquito predators were identified from the larval habitats, including Hydrophilidae, Dytiscidae, Corixidae, Nepidae, Notonectidae, Belostomatidae, and Cordulidae. Predator density in disused goldmines was significantly higher than that of other habitat types. Determination of the relative importance of predation, habitat stability and food contents on natural mosquito habitat productivity would help to design cost-effective vector control methods specifically targeted at the productive habitats.

Information on the spatial relationships between disease vectors and environmental factors is fundamental to vector-borne disease control. Although it is well known that mosquito abundance is associated with the amount of rainfall and thus the number of larval breeding sites, the spatial relationship between larval habitat availability and adult mosquito abundance is not clear. We investigated the impact of environmental heterogeneity and larval habitats on the spatial distribution of Anopheles gambiae s. s. and An. funestus adult mosquitoes, the most important malaria vectors in the highlands of western Kenya. Mosquito sampling was conducted in May, August, and November 2002, and February 2003. Geographic information system layers of larval habitats, land use type, human population distribution, house structure, and hydrologic schemes were overlaid with adult mosquito abundance. Correlography was used to determine the spatial autocorrelation in adult mosquito abundance among houses and the cross-correlation between adult mosquito abundance and environmental factors. Getis' G(i)(*)(d) index was used to define focal adult mosquito abundance clusters. We found a significant autocorrelation in the vector population and a significant cross-correlation between the vector population and larval habitat availability. The threshold distances of both autocorrelation and cross-correlation were significantly varied among seasons. Focal clustering analysis revealed that the adult vector population was concentrated along the Yala River Valley where most larval habitats were found. Regression analysis found that distance of a house to the Yala River, age of the house, elevation, house structure, and tree canopy coverage significantly affected adult mosquito abundance. Our results suggest that vector control targeted at malaria transmission hotspots and supplemented by larval control may be an effective approach for epidemic malaria control in the western Kenya highlands.

The reasons for the resurgence of malaria in the African highlands have been subject of debate. Because vector abundance is important for malaria transmission, gaining a better understanding of vector biology is a key to understanding the mechanisms of highland malaria. We studied vector productivity in relation to the highland environment and compared productivity between lowland and highland sites. We found lower vector productivity in the highland and in wetlands where the temperature was lower. Immature stage development time was significantly longer in the highland site. Development time was significantly shorter in aquatic habitats in cultivated areas than in wetlands, and survival rate was significantly higher in cultivated areas. Fecundity was significantly lower in the highland site. These findings suggest that changes in local temperature and land use contribute to an increase of malaria vectors in the highland.

We examined whether predators and competitors influence selection of oviposition sites by Anopheles gambiae Giles. Mosquitoes in cages laid significantly fewer eggs in rainwater conditioned with a predator (backswimmers, Notonecta sp.) than in unconditioned rainwater. Rainwater conditioned with a putative competitor (tadpoles, Xenopus sp.) also had fewer eggs than unconditioned rainwater. Similarly, mosquitoes laid significantly fewer eggs in rainwater conditioned with five and 50 An. gambiae larvae than in unconditioned rainwater. When larvae were present, significantly more eggs were laid in containers with five larvae than in containers with higher densities, but the differences in number of eggs laid were not significant among the densities of 40, 70, and 100 larvae. This study demonstrated that caged An. gambiae females avoid oviposition in habitats with supposed competitors and predators.

We examined the effects of land cover type on survivorship and productivity of Anopheles gambiae in Kakamega in the western Kenyan highlands (elevation = 1,420-1,580 meters above sea level). Under natural conditions, An. gambiae sensu lato adults emerged only from farmland habitats, with an estimated productivity of 1.82 mosquitoes/meter(2)/week, but not from forest and swamp habitats. To determine the effects of intraspecific competition and land cover types, semi-natural larval habitats were created within three land cover types (farmland, forest, and natural swamp), and three different densities of An. gambiae sensu stricto larvae were introduced to the larval habitats. The mosquito pupation rate in farmland habitats was significantly greater than in swamp and forest habitats, and larval-to-pupal development times were significantly shorter. At higher densities, the larvae responded to increased intraspecific competition by extending their development time and emerging as smaller adults, but initial larval density showed no significant effects on pupation rate. Land cover type may affect larval survivorship and adult productivity through its effects on water temperature and nutrients in the aquatic habitats, as shown by the significantly higher water temperature in farmland habitats, enhanced pupation rates and shortened development times from the addition of food to habitats, and a significant negative correlation of the occurrence of An. gambiae larvae with canopy cover and emergent plants in natural habitats. These results suggest that deforestation and cultivation of natural swamps in the western Kenyan highland create conditions favorable for the survival of An. gambiae larvae, and consequently increase the risks of malaria transmission to the human population.

We studied the oviposition site preference and egg hatchability of Anopheles gambiae Giles with water collected from farmlands, forests, and natural wetlands. Water types significantly affected oviposition preference. Mosquitoes deposited significantly more eggs in rainwater in both the dry and wet seasons than waters from forests and wetlands, suggesting that An. gambiae prefers water with few impurities for oviposition. In the dry season, An. gambiae females also deposited significantly more eggs in waters from farmlands than those from forests and natural wetlands, but these differences were not statistically significant during the wet season. In both indoor and natural conditions, egg mortality in natural wetland habitats was significantly higher than in farmland habitats. The average water temperature in natural wetland habitats was significantly lower than farmland habitats in the natural conditions, but it remained the same under indoor experimental conditions, suggesting that factors other than water temperature play an important role in egg hatchability. Together with the findings from previous studies on the effects of land cover on larval survivorship, our results support the hypothesis that variations in habitat conditions induced by different land cover types contribute to the heterogeneous spatial distribution of An. gambiae larvae in the western Kenya highland.

We carried out a population-based study to determine the unbiased, age-specific Plasmodium falciparum prevalence, asexual and sexual parasite density, and spatial distribution to establish rates of infection at a site in western Kenya. Three cross-sectional surveys were carried out in western Kenya highlands. Blood samples were taken from 1,388 persons from 6 months to 75 years of age. Parasite prevalence and densities in the population decreased with age and distance from valley bottoms. Children from 1 to 4 years of age had the highest parasite prevalence (38.8%-62.8%); in adults, prevalence declined to 2.9%-24.1%. Malaria prevalence declined by an average of 19% from July to December 2002 across age groups. These observations suggest that parasite transmission is intense at this altitude. Asexual parasite density indicated clustering near major vector breeding habitats. Variability in seasonal prevalence indicates transmission instability and susceptibility to epidemics.

The distributions of anopheline larval habitats were aggregated in valley bottoms in Kenya in both the rainy and dry seasons, although the degree of aggregation was higher in the dry season than in the rainy season. Larvae of the Anopheles gambiae complex larvae were found more frequently in habitats in farmlands and pastures. However, An. funestus larvae were found more frequently in natural swamps and pastures. Canopy cover was the only variable significantly associated with the occurrence of the An. gambiae complex and An. funestus. The average canopy cover was significantly less in the habitats with the An. gambiae complex and An. funestus larvae than those without the anopheline larvae. Thus, land cover types and topographic features showed important effects on the distribution of anopheline larval habitats. These results suggest that clearing riparian forests would improve growing conditions of the An. gambiae complex and An. funestus larvae in Kenyan highlands.

The causes of the recent re-emergence of malaria in the East African highlands probably involve a complex interplay among multiple factors, including climate, land use, topography, inadequate use of antimalarial drugs and drug resistance, socioeconomic status, health policy and public health control measures. It is important to determine the relative contribution of these factors. In our study, we statistically attributed the effects of autocorrelation, seasonality and climate variability to the temporal variation in the number of malaria patients in several highland sites in East Africa. We found that in three out of seven sites, climate variability contributed more variance to malaria patient numbers than did autocorrelation and seasonality. In all seven study sites, we found highly significant nonlinear, synergistic effects of the interaction between rainfall and temperature on malaria patient time series.

BackgroundUnderstanding the genetic structure of island Anopheles gambiae populations is important for the current tactics in mosquito control and for the proposed strategy using genetically-modified mosquitoes (GMM). Genetically-isolated mosquito populations on islands are a potential site for testing GMM. The objective of this study was to determine the genetic structure of A. gambiae populations on the islands in Lake Victoria, western Kenya.MethodsThe genetic diversity and the population genetic structures of 13 A. gambiae populations from five islands on Lake Victoria and six villages from the surrounding mainland area in the Suba District were examined using six microsatellite markers. The distance range of sampling sites varied between 2.5 and 35.1 km.ResultsA similar level of genetic diversity between island mosquito populations and adjacent mainland populations was found. The average number of alleles per locus was 7.3 for the island populations and 6.8 for the mainland populations. The average observed heterozygosity was 0.32 and 0.28 for the island and mainland populations, respectively. A low but statistically significant genetic structure was detected among the island populations (FST = 0.019) and between the island and mainland populations (FST = 0.003). A total of 12 private alleles were found, and nine of them were from the island populations.ConclusionA level of genetic differentiation between the island and mainland populations was found. Large extent of gene flow between the island and mainland mosquito populations may result from wind- or human-assisted dispersal. Should the islands on Lake Victoria be used as a trial site for the release program of GMM, mosquito dispersal between the islands and between the island and the mainland should be vigorously monitored.

The current study examined temporal and spatial distribution patterns of anopheline malaria vectors in a highland site and determined the number of houses to be sampled to achieve the targeted precision level. Adult mosquito sampling was conducted seasonally in May and August 2002 in a 3 by 3-km2 area, and in November 2002 and February 2003 in an expanded 4 by 4-km2 area in Kakamega District, western Kenya. Anopheles gambiae Giles was the predominant malaria vector species, constituting 84.6% of the specimens, whereas Anopheles funestus Giles constituted 15.4% of the vector populations. An. gambiae abundance increased by six- to eight-fold in the long rainy season over the dry seasons, but An. funestus abundance peaked 3 mo after the long rainy season. For both species, the coefficient of variation was larger than 1, suggesting that the distribution of mosquito adults was aggregated. Mosquito clustering occurred in houses