Susceptibility of Anopheles gambiae complex mosquitoes to microbial larvicides in diverse ecological settings in western Kenya.

The microbial larvicides Bacillus thuringiensis var. israelensis (Bti) and Bacillus sphaericus (Bs) (Bacillales: Bacillaceae) are well known for their efficacy and safety in mosquito control. In order to assess their potential value in future mosquito control strategies in western Kenya, the current study tested the susceptibility of five populations of Anopheles gambiae complex mosquitoes (Diptera: Culicidae), collected from five diverse ecological sites in this area, to Bti and Bs under laboratory conditions. In each population, bioassays were conducted with eight concentrations of larvicide (Bti/Bs) in four replicates and were repeated on three separate days. Larval mortality was recorded at 24 h or 48 h after the application of larvicide and subjected to probit analysis. A total of 2400 An. gambiae complex larvae from each population were tested for their susceptibility to Bti and Bs. The mean (± standard error of the mean, SEM) lethal concentration values of Bti required to achieve 50% and 95% larval mortality (LC50 and LC95 ) across the five populations were 0.062 (± 0.005) mg/L and 0.797 (± 0.087) mg/L, respectively. Corresponding mean (± SEM) values for Bs were 0.058 (± 0.005) mg/L and 0.451 (± 0.053) mg/L, respectively. Statistical analysis indicated that the five populations of An. gambiae complex mosquitoes tested were fully susceptible to Bti and Bs, and there was no significant variation in susceptibility among the tested populations.

Early biting of the Anopheles gambiae s.s. and its challenges to vector control using insecticide treated nets in western Kenya highlands.

Long term use of insecticides in malaria vector control has been shown to alter the behavior of vectors. Such behavioral shifts have the potential of undermining the effectiveness of insecticide-based control interventions. The effects of insecticide treated nets (ITNs) use on the composition, biting/feeding and sporozoite rates of Anopheles gambiae s.l. mosquitoes in Musilongo village, Vihiga County of western Kenya highlands were investigated. Adult mosquitoes were collected in selected sleeping spaces inside six randomly selected houses using miniature Centre for Disease Control and Prevention (CDC) light traps. Mosquito sampling in each house was conducted twice every week for 16 consecutive months (May 2010-August 2012). At each sampling a single trap was set in the selected space inside each house such that it collected mosquitoes alternatively from 18:00 to 21:00h and 21:00 to 06:00h every week. All collected mosquitoes were morphologically identified. Female Anopheles mosquitoes were classified according to their physiological status as unfed, fed, partially gravid and gravid, sorted and counted. Members of the A. gambiae complex were identified using a Polymerase chain reaction (PCR) method. Enzyme-linked-immunosorbent assay (ELISA) was used to determine blood meal sources and Plasmodium infection rates in A. gambiae s.l. mosquitoes. Blood meal tests were conducted on DNA extracted from gut contents of blood fed A. gambiae s.l. The head and thorax section of dried samples of A. gambiae s.l. were used in testing for the presence of Plasmodium falciparum (Pf) sporozoites. Overall, 735 adult female Anopheles comprising 708 [96.3%] A. gambiae s.l. and 27 [3.7%] Anopheles funestus mosquitoes were collected. A. gambiae s.l. population collected comprised, 615 [86.9%] unfed and 38 [5.4%] fed adult mosquitoes. The rest were either partially or fully gravid. The proportion of A. gambiae s.l. biting indoors within 18:00-21:00h was 15.8% (103/653) at a rate of 3.2bites per person per hour compared to 84.2% biting from 21:00-06:00h at a rate of 3.8 bites/per/h. An estimated 97.7% A. gambiae ss and 2.3% A. arabiensis constituted the indoor biting A. gambiae s.l. The population of An. gambiae s.l. biting from 18:00 to 21:00h had a Plasmodium faciparum (pf) sporozoite rate of 3.8% compared to 3.5% observed in populations biting within 21:00-06:00h. Human blood constituted 89% of An. gambiae s.l. blood meal sources. The risk of malaria transmission from 21:00 to 06:00h was approximately 5 fold the risk within 18:00-21:00h. Majority of the infective female A. gambiae s.l. adults were biting deep into the night than in the early hours of the night. Humans remain the preferred source of blood meal for A. gambiae s.s. the dominant malaria vector in the highlands. ITNs remain a fundamental control intervention against malaria transmission since female blood seekers were more during bed time than pre-bed time. Advocacy on enhanced net availability, integrity and usage in Kenyan highlands can reduce Pf transmission. Additional complementary interventions are required to control the biting and parasite transmission encountered before bed-time.

The reservoir of Plasmodium falciparum malaria in a holoendemic area of western Kenya.

The reservoir of infectious Plasmodium falciparum gametocytes in a population living in an area of holoendemic malaria in western Kenya was estimated by directly feeding mosquitoes on volunteers. Resulting mosquito infections were assessed both by midgut examination for oocysts and by enzyme-linked immunosorbent assay for P. falciparum circumsporozoite antigen. Calculations based on the age structure of the population and the resulting rates of mosquito infections indicated that children under 10 years of age were responsible for 72% of mosquito infections, individuals between 10 and 21 years of age contributed 12%, and those over 21 years of age accounted for 16%. No infection resulted in mosquitoes fed on infants less than 1 year of age.

Small-scale field evaluation of the monomolecular surface film 'Arosurf MSF' against Anopheles arabiensis Patton.

A field trial was conducted to test the insecticidal action of the monolayer surface film 'Arosurf MSF' applied by knapsack sprayers, against larvae and pupae of Anopheles arabiensis Patton in a rice irrigation scheme in Western Kenya. Larval and pupal densities and the number of emerging adults were determined by dipping and emergence cages respectively. Application of the monolayer by knapsack sprayers provided good coverage. There were high daily mortalities of the fourth instar larvae, with no adult emergence from 'Arosurf MSF' treated plots compared to lower fourth instar mortalities and continuous adult emergence from untreated control plots, indicating the potential of the monolayer for control of An. arabiensis mosquitoes in rice fields.

The effects of rainfall and evapotranspiration on the temporal dynamics of Anopheles gambiae s.s. and Anopheles arabiensis in a Kenyan village.

The population dynamics of the larval and adult life stages of the malaria vector Anopheles gambiae Giles were studied in Miwani, western Kenya, in relation to meteorological conditions. Larval density within a habitat, the number of larval habitats and sibling species composition were investigated as determinants of larval population dynamics. Female vector densities inside local houses and sibling species composition were investigated as determinants of adult population dynamics. Larval densities were estimated using a modified area-sampling method. Within the habitats, all instars showed a highly aggregated distribution, with the exception of second instars. A longitudinal study on the larval populations of A. gambiae s.l. in two different types of habitat (dirt track and ditch) was carried out, using a novel sampling procedure. A. gambiae s.s. and Anopheles arabiensis, the two sibling species occurring sympatrically in the study area, showed some spatial segregation between the two types of habitat. Rainfall was significantly correlated with the number of A. gambiae s.l. larval habitats during the first 6 weeks of study taking 1 week time lag into account, while over the entire 5-month study period correlations were less clear. With 1 week time lag, rainfall was also significantly correlated with the number of female A. gambiae s.l. collected from CDC-light traps in the study houses. Both larval and adult populations showed a significant increase in the proportion of A. gambiae s.s. within the mixed population of A. gambiae s.s. and A. arabiensis over time. Although not significantly correlated, the ratio of rainfall over precipitation/potential evapotranspiration (P/PE), indicative of the humidity conditions in the area, was probably the driving force of this increase.

Origin of blood meals in indoor and outdoor resting malaria vectors in western Kenya.

Blood meals were obtained from indoor and outdoor resting malaria vectors in three villages of western Kenya and tested by sandwich ELISA to determine host preferences and their human blood index (HBI). Anopheles gambiae s.s. collected indoors at Kisian village had a HBI of 0.97 while that of Anopheles arabiensis collected at Ahero was 0.23. However, the HBI of A. arabiensis varied depending on the availability of outdoor resting shelters. Most female A. arabiensis (98.9%) collected outdoors in granaries at Ahero had fed on cattle. Indoor-collected female Anopheles funestus had mainly fed on people (93.0%), but taken at least some of their blood (20.2%) from cattle. Although small numbers of A. arabiensis fed on sheep or goats and birds, none of the female A. gambiae s.s. and A. funestus tested had fed on these hosts. The absence of human-fed A. arabiensis in outdoor shelters indicated that exiting after feeding, a behaviour pattern that mitigates indoor insecticidal spraying, is not prevalent in this species in western Kenya.

Effect of human circumsporozoite antibodies in Plasmodium-infected Anopheles (Diptera: Culicidae).

Human circumsporozoite (CS) antibodies to Plasmodium falciparum were detected in blood meals from 45.0% of 1,547 field-collected Anopheles gambiae Giles sensu lato and Anopheles funestus Giles from western Kenya. Possible effects on malaria infections within the Anopheles host were investigated. Circumsporozoite antibodies were detected in blood meals up to 36 h after feeding. Antibodies crossing the midgut were detected experimentally in hemolymph from 4 to 36 h after feeding; human IgG also was present in hemolymph from fully gravid field-collected Anopheles. Ingestion of high-titer human CS antibodies or 2A10 monoclonal antibody to P. falciparum sporozoites by P. falciparum-infected An. gambiae, 10 d after feeding on an infected human, had no effect on oöcyst maturation, sporozoite rates, or sporozoite loads. Contact between CS antibodies and sporozoites in the hemocoel did not block sporozoite invasion of salivary glands. Human IgG antibodies were detected by an indirect fluorescent antibody technique on salivary gland sporozoites from 83.3% of 114 field-collected Anopheles. In 65.4% of 26 infections, antibodies persisted on sporozoites for at least three days. Thus, a high proportion of naturally infected An. gambiae s.l. and An. funestus in western Kenya transmit sporozoites that are bound with human IgG acquired during previous blood meals. The infectivity of such sporozoites needs to be determined in relation to natural transmission and to the potential use of malaria sporozoite vaccines.

Resting behaviour, ecology and genetics of malaria vectors in large scale agricultural areas of Western Kenya.

In Kenya indoor and outdoor resting densities of Anopheles arabiensis and Anopheles funestus at the Ahero rice irrigation scheme, and Anopheles gambiae s.s., An. arabiensis and An. funestus at the Miwani sugar belt were assessed for 13 months by pyrethrum spray collections in houses and granaries. The vector's house leaving behaviour was evaluated with exit traps and it was noted that early exophily (i.e., deliberate) was not detected in any of the vectors. Assortative indoor/outdoor resting behaviour was studied by a capture-mark-release-recapture method and showed that in An. arabiensis both indoor and outdoor resting traits were present in the same individuals. Samples of half-gravid female An. gambiae s.l. were chromosomally identified either as Anopheles gambiae s.s. or An. arabiensis and in a subsample chromosomal inversions were read. Anopheles gambiae s.s. and An. arabiensis had the 2Rb inversion but in addition the 2La inversion was found in An. gambiae s.s. and this is an indication of low chromosomal variation. At Ahero An. arabiensis was most abundant when the rice crop was immature and An. funestus when the crop was mature. This succession of vectors facilitated the transmission of malaria throughout the year. At Miwani, An. gambiae s.l. population peaked during the long rains but the proportion of An. arabiensis was highest during the dry season. The indoor resting density of males of the three vector species was less than half of the females.

Field evaluation of arthropod repellents, deet and a piperidine compound, AI3-37220, against Anopheles funestus and Anopheles arabiensis in western Kenya.

A field evaluation of the repellents N,N-diethyl-3-methylbenzamide (deet) and 1-(3-cyclohexen-1-yl-carbonyl)-2-methylpiperidine (AI3-37220, a piperidine compound) was conducted against Anopheles funestus and An. arabiensis in Kenya. Both repellents provided significantly more protection (P < 0.001) than the ethanol control. AI3-37220 was significantly more effective (P < 0.001) than deet in repelling both species of mosquitoes. After 9 h, 0.1 mg/cm2 of AI3-37220 provided 89.8% and 71.1% protection against An. arabiensis and An. funestus, respectively. Deet provided > 80% protection for only 3 h, and protection rapidly decreased after this time to 60.2% and 35.1% for An. arabiensis and An. funestus, respectively, after 9 h. Anopheles funestus was significantly less sensitive (P < 0.001) to both repellents than An. arabiensis. The results of this study indicate that AI3-37220 is more effective than deet in repelling anopheline mosquitoes in western Kenya.

Abdominal cocoon: report of five cases.

Abdominal cocoon (idiopathic sclerosing peritonitis) has now been reported in many parts of the world but remains principally a disease of the tropics and sub-tropics. Five cases diagnosed at a rural Kenyan hospital between 1993 and 1998 are presented and the operative findings and management is discussed. Although the condition normally occurs in adolescent girls, one patient was noted to be forty two years of age, and was subsequently found to have AIDS.

Knowledge and attitudes to malaria control and acceptability of permethrin impregnated sisal curtains.

OBJECTIVES: The objectives of this study were to conduct a survey on the knowledge and practices relating to malaria prevention and treatment in two rural communities in Western Kenya, and to determine the acceptability of specially designed permethrin impregnated sisal strands curtains previously introduced into one of the study communities as barriers to mosquito biting. DESIGN: A knowledge, attitudes and practices (KAP) survey was conducted by pre-trained interviewers using a pre-tested questionnaire. SETTING: The study was conducted in two communities located 15 km northwest of Kisumu town, and next to the swamps bordering Lake Victoria in Kisumu District, Western Kenya. PARTICIPANTS: Adult individuals from 50 houses selected from the intervention, and 50 houses from the control sites were included in the study. RESULTS: Both communities had a clear conception of malaria and its symptoms, and of the mosquito as its vector. Malaria was recognised as a potential cause of death by 44% and 72% of the participants in the intervention and control sites respectively. Sixty two per cent to 74% of the people interviewed in both sites named mosquitoes as the causative agents of malaria. Chloroquine was still the drug of choice for malaria treatment, with over 70% usage among the study population. Mention of pyrimethamine/sulfadoxine and pyrimethamine/sulfalene as alternative therapy was below five per cent. Despite a high level (86%) of awareness of bednets as effective barriers to mosquito biting, they were reported in use by less than 35% in both communities. Sisal strand curtains were considered effective and acceptable to more than 80% of the community. CONCLUSIONS: The results of this study underscore the existing need for affordable means of mosquito control, such as sisal strand curtains, for such rural communities which may be acutely aware of the problems associated with malaria, but are constrained from taking any action by lack of resources.

The effect of permethrin impregnated sisal curtains on vector density and malaria incidence: a pilot study.

Impregnation of bednets and curtains with suitable pyrethroids may reduce entomological inoculation rates (EIR) and malaria incidence. We conducted a quasi-experimental pilot study over 3 months in Western Kenya on 20 houses with 54 children. Ten houses in the experimental site received sisal curtains treated with permethrin at either 0.5g/m2 or 0.1g/m2. Control houses had untreated curtains or none. Mosquito vector density (MVD), man biting rates (MBR), and residual insecticidal effects (RIE) of permethrin were determined every two weeks. MVD was reduced by 97.7% and 98.7% in houses from the 2 experimental groups with a 60% reduction with unimpregnated curtains. MBR varied from 6.4 (no curtains), 1.7 (unimpregnated), 0.7 (0.5g/m2 curtains) to 0.4 (1.0g/m2 curtains). RIE begun to decline after the fourth month. Malaria incidence remained similar at the two sites. We conclude that covering of eaves and windows with permethrin impregnated sisal curtains can reduce MVD and the number of mosquito bites to individuals sleeping in protected houses.

Progress towards understanding the ecology and epidemiology of malaria in the western Kenya highlands: opportunities and challenges for control under climate change risk.

Following severe malaria epidemics in the western Kenya highlands after the late 1980s it became imperative to undertake eco-epidemiological assessments of the disease and determine its drivers, spatial-temporal distribution and control strategies. Extensive research has indicated that the major biophysical drivers of the disease are climate change and variability, terrain, topography, hydrology and immunity. Vector distribution is focalized at valley bottoms and abundance is closely related with drainage efficiency, habitat availability, stability and productivity of the ecosystems. Early epidemic prediction models have been developed and they can be used to assess climate risks that warrant extra interventions with a lead time of 2-4 months. Targeted integrated vector management strategies can significantly reduce the cost on the indoor residual spraying by targeting the foci of transmission in transmission hotspots. Malaria control in the highlands has reduced vector population by 90%, infections by 50-90% in humans and in some cases transmission has been interrupted. Insecticide resistance is increasing and as transmission decreases so will immunity. Active surveillance will be required to monitor and contain emerging threats. More studies on eco-stratification of the disease, based on its major drivers, are required so that interventions are tailored for specific ecosystems. New and innovative control interventions such as house modification with a one-application strategy may reduce the threat from insecticide resistance and low compliance associated with the use of ITNs.

Variation in malaria transmission dynamics in three different sites in Western kenya.

The main objective was to investigate malaria transmission dynamics in three different sites, two highland villages (Fort Ternan and Lunyerere) and a lowland peri-urban area (Nyalenda) of Kisumu city. Adult mosquitoes were collected using PSC and CDC light trap while malaria parasite incidence data was collected from a cohort of children on monthly basis. Rainfall, humidity and temperature data were collected by automated weather stations. Negative binomial and Poisson generalized additive models were used to examine the risk of being infected, as well as the association with the weather variables. Anopheles gambiae s.s. was most abundant in Lunyerere, An. arabiensis in Nyalenda and An. funestus in Fort Ternan. The CDC light traps caught a higher proportion of mosquitoes (52.3%) than PSC (47.7%), although not significantly different (P = 0.689). The EIR's were 0, 61.79 and 6.91 bites/person/year for Fort Ternan, Lunyerere and Nyalenda. Site, month and core body temperature were all associated with the risk of having malaria parasites (P < 0.0001). Rainfall was found to be significantly associated with the occurrence of P. falciparum malaria parasites, but not relative humidity and air temperature. The presence of malaria parasite-infected children in all the study sites provides evidence of local malaria transmission.

The effect of water turbidity on the near-surface water temperature of larval habitats of the malaria mosquito Anopheles gambiae.

Water temperature is an important determinant in many aquatic biological processes, including the growth and development of malaria mosquito (Anopheles arabiensis and A. gambiae) immatures. Water turbidity affects water temperature, as suspended particles in a water column absorb and scatter sunlight and hence determine the extinction of solar radiation. To get a better understanding of the relationship between water turbidity and water temperature, a series of semi-natural larval habitats (diameter 0.32 m, water depth 0.16 m) with increasing water turbidity was created. Here we show that at midday (1300 hours) the upper water layer (thickness of 10 mm) of the water pool with the highest turbidity was on average 2.8 degrees C warmer than the same layer of the clearest water pool. Suspended soil particles increase the water temperature and furthermore change the temperature dynamics of small water collections during daytime, exposing malaria mosquito larvae, which live in the top water layer, longer to higher temperatures.

Low larval vector survival explains unstable malaria in the western Kenya highlands.

Several highland areas in eastern Africa have recently suffered from serious malaria epidemics. Some models predict that, in the short term, these areas will experience more epidemics as a result of global warming. However, the various processes underlying these changes are poorly understood. We therefore investigated malaria prevalence, malaria vector densities and malaria vector survival in a highland area in western Kenya, ranging from approximately 1,550-1,650 m altitude. Although only five adult malaria vectors were collected during 180 light traps and 180 resting collections over a 23-month study period, malaria was prevalent among school children (average parasite prevalence: 10%). During an extensive survey of potential larval habitats, we identified only seven habitats containing Anopheles gambiae Giles s.l. larvae. Their limited number and low larval densities suggested that their contribution to the adult vector population was small. Experiments on adult and larval survival showed that at this altitude, adult mosquitoes survived inside local houses, but that larval development was severely retarded: only 2 of 500 A. gambiae s.l. larvae developed to the pupal stage, whereas all other larvae died prior to pupation. At present, high vector densities are unlikely because of unfavourable abiotic conditions in the area. However, temporary favourable conditions, such as during El Niño years, may increase larval vector survival and may lead to malaria epidemics.

Climate change and vector-borne diseases: a regional analysis.

Current evidence suggests that inter-annual and inter-decadal climate variability have a direct influence on the epidemiology of vector-borne diseases. This evidence has been assessed at the continental level in order to determine the possible consequences of the expected future climate change. By 2100 it is estimated that average global temperatures will have risen by 1.0-3.5 degrees C, increasing the likelihood of many vector-borne diseases in new areas. The greatest effect of climate change on transmission is likely to be observed at the extremes of the range of temperatures at which transmission occurs. For many diseases these lie in the range 14-18 degrees C at the lower end and about 35-40 degrees C at the upper end. Malaria and dengue fever are among the most important vector-borne diseases in the tropics and subtropics; Lyme disease is the most common vector-borne disease in the USA and Europe. Encephalitis is also becoming a public health concern. Health risks due to climatic changes will differ between countries that have developed health infrastructures and those that do not. Human settlement patterns in the different regions will influence disease trends. While 70% of the population in South America is urbanized, the proportion in sub-Saharan Africa is less than 45%. Climatic anomalies associated with the El Niño-Southern Oscillation phenomenon and resulting in drought and floods are expected to increase in frequency and intensity. They have been linked to outbreaks of malaria in Africa, Asia and South America. Climate change has far-reaching consequences and touches on all life-support systems. It is therefore a factor that should be placed high among those that affect human health and survival.

Field testing an enzyme-linked synthetic oligonucleotide probe for identification of Anopheles gambiae s.s. and An. arabiensis.

Tests were carried out in Kenya to determine whether the enzyme-linked synthetic oligonucleotide probe (pAna 1) developed for identifying species of the Anopheles gambiae complex could be used under field conditions. The An. arabiensis male-specific pAna 1 probe was able to identify all male larval instars and adult males. However, the non-radioactive assay was not sufficiently sensitive to identify male sperm DNA in all the mated female An. arabiensis. Although the ratio of An. arabiensis to An. gambiae s.s. identified with pAna 1 in males during the dry season was in agreement with the ratio in half-gravid females identified cytogenetically, the ratios were different during the wet season. This study demonstrates that the enzyme-linked DNA probe assay is applicable under field conditions.

Plasmodium falciparum sporozoite and entomological inoculation rates at the Ahero rice irrigation scheme and the Miwani sugar-belt in western Kenya.

Anopheles arabiensis and An. funestus were collected by pyrethrum spray sheet collections in houses and by human-bait catches at a village in western Kenya adjacent to the Ahero rice irrigation scheme; and using the same methods, An. gambiae s.l. and An. funestus were collected at Miwani, a village in the sugar-cane belt. Plasmodium falciparum sporozoite rates were determined by ELISA. At Ahero the mean sporozoite rates were 1.1% and 4.3% in An. arabiensis and An. funestus, respectively, while at Miwani the rates were 6.0% in An. gambiae s.l. and 4.3% in An. funestus. Entomolgoical inoculation rates (EIR) were derived from both human-bait collections (IR-HBC) and by the proportion of human blood-fed females caught resting indoors (IR-HBF). The IR-HBF appeared to be a more realistic index of EIR. At Ahero and Miwani people were exposed to an average of 416 and 91 infective bites/person/year, respectively. The main vectors were An. funestus at Ahero and An. gambiae s.l. at Miwani. In view of the intense and perennial malaria transmission at Ahero, vector control by insecticides should be considered, while at Miwani, where transmission is seasonal, permethrin-impregnated bed nets could be an alternative to indoor spraying. These measures must be augmented with availability of effective antimalarials.