A field bioassay for assessing ivermectin bio-efficacy in wild malaria vectors.

BACKGROUND: Ivermectin (IVM) mass drug administration is a candidate complementary malaria vector control tool. Ingestion of blood from IVM treated hosts results in reduced survival in mosquitoes. Estimating bio-efficacy of IVM on wild-caught mosquitoes requires they ingest the drug in a blood meal either through a membrane or direct feeding on a treated host. The latter, has ethical implications, and the former results in low feeding rates. Therefore, there is a need to develop a safe and effective method for IVM bio-efficacy monitoring in wild mosquitoes. METHODS: Insectary-reared Anopheles gambiae s.s. were exposed to four IVM doses: 85, 64, 43, 21 ng/ml, and control group (0 ng/ml) in three different solutions: (i) blood, (ii) 10% glucose, (iii) four ratios (1:1, 1:2, 1:4, 1:8) of blood in 10% glucose, and fed through filter paper. Wild-caught An. gambiae s.l. were exposed to 85, 43 and 21 ng/ml IVM in blood and 1:4 ratio of blood-10% glucose mixture. Survival was monitored for 28 days and a pool of mosquitoes from each cohort sacrificed immediately after feeding and weighed to determine mean weight of each meal type. RESULTS: When administered in glucose solution, mosquitocidal effect of IVM was not comparable to the observed effects when similar concentrations were administered in blood. Equal concentrations of IVM administered in blood resulted in pronounced reductions in mosquito survival compared to glucose solution only. However, by adding small amounts of blood to glucose solution, mosquito mortality rates increased resulting in similar effects to what was observed during blood feeding. CONCLUSION: Bio-efficacy of ivermectin is strongly dependent on mode of drug delivery to the mosquito and likely influenced by digestive processes. The assay developed in this study is a good candidate for field-based bio-efficacy monitoring: wild mosquitoes readily feed on the solution, the assay can be standardized using pre-selected concentrations and by not involving treated blood hosts (human or animal) variation in individual pharmacokinetic profiles as well as ethical issues are bypassed. Meal volumes did not explain the difference in the lethality of IVM across the different meal types necessitating further research on the underlying mechanisms.

Malaria vector bionomics and transmission in irrigated and non-irrigated sites in western Kenya.

Irrigation not only helps to improve food security but also creates numerous water bodies for mosquito production. This study assessed the effect of irrigation on malaria vector bionomics and transmission in a semi-arid site with ongoing malaria vector control program. The effectiveness of CDC light traps in the surveillance of malaria vectors was also evaluated relative to the human landing catches (HLCs) method. Adult mosquitoes were sampled in two study sites representing irrigated and non-irrigated agroecosystems in western Kenya using a variety of trapping methods. The mosquito samples were identified to species and assayed for host blood meal source and Plasmodium spp. sporozoite infection using polymerase chain reaction. Anopheles arabiensis was the dominant malaria vector in the two study sites and occurred in significantly higher densities in irrigated study site compared to the non-irrigated study site. The difference in indoor resting density of An. arabiensis during the dry and wet seasons was not significant. Other species, including An. funestus, An. coustani, and An. pharoensis, were collected. The An. funestus indoor resting density was 0.23 in irrigated study site while almost none of this species was collected in the non-irrigated study site. The human blood index (HBI) for An. arabiensis in the irrigated study site was 3.44% and significantly higher than 0.00% for the non-irrigated study site. In the irrigated study site, the HBI of An. arabiensis was 3.90% and 5.20% indoor and outdoor, respectively. The HBI of An. funestus was 49.43% and significantly higher compared to 3.44% for An. arabiensis in the irrigated study site. The annual entomologic inoculation rate for An. arabiensis in the irrigated study site was 0.41 and 0.30 infective bites/person/year indoor and outdoor, respectively, whereas no transmission was observed in the non-irrigated study site. The CDC light trap performed consistently with HLC in terms of vector density. These findings demonstrate that irrigated agriculture may increase the risk of malaria transmission in irrigated areas compared to the non-irrigated areas and highlight the need to complement the existing malaria vector interventions with novel tools targeting the larvae and both indoor and outdoor biting vector populations.

Larval density dependence in Anopheles gambiae s.s., the major African vector of malaria.

Anopheles gambiae sensu stricto is the most important vector of malaria in Africa although relatively little is known about the density-dependent processes determining its population size. Mosquito larval density was manipulated under semi-natural conditions using artificial larval breeding sites placed in the field in coastal Kenya; two experiments were conducted: one manipulating the density of a single cohort of larvae across a range of densities and the other employing fewer densities but with the treatments crossed with four treatments manipulating predator access. In the first experiment, larval survival, development rate and the size of the adult mosquito all decreased with larval density (controlling for block effects between 23% and 31% of the variance in the data could be explained by density). In the second experiment, the effects of predator manipulation were not significant, but again we observed strong density dependence in larval survival (explaining 30% of the variance). The results are compared with laboratory studies of A. gambiae larval competition and the few other studies conducted in the field, and the consequences for malaria control are discussed.

Malaria vector bionomics in Taita-Taveta County, coastal Kenya.

BACKGROUND: Estimation of the composition and densities of mosquito species populations is crucial for monitoring the epidemiology of mosquito-borne diseases and provide information on local vectors to public health officials and policy-makers. The aim of this study was to evaluate malaria vector bionomics in ecologically distinct sites in Taita-Taveta County, Kenya. METHODS: Adult mosquitoes were collected using backpack aspirators and paired indoor/outdoor CDC light traps in 10 randomly selected households in six villages with distinct ecologies over a study period of 3 years. All Anopheles mosquitoes were morphotyped, and sibling species of Anopheles gambiae sensu lato (An. gambiae s.l.) were identified and separated by PCR analysis of extracted ribosomal DNA. All female anophelines were tested for sporozoite infectivity, with engorged females screened for blood-meal sources using the enzyme-linked immunosorbent assay technique. A subsample of those testing positive and those testing negative for Plasmodium in the ELISA were subjected to PCR assay. RESULTS: A total of eight different Anopheles species were collected both indoors and outdoors. Anopheles gambiae s.l. (82.6%, n = 5252) was the predominant species sensu lato, followed by Anopheles coustani sensu lato (An. coustani s.l.; (10.5%, n = 666) and Anopheles funestus sensu lato (An. funestus s.l.; 5.6%, n = 357). A subset of 683 mosquito samples representing An. gambiae s.l. (n = 580, approx. 11.0%) and An. funestus s.l. (n = 103, approx. 28.9%) were identified by molecular diagnostic assays into sibling species. The An. gambiae s.l. complex was composed of Anopheles arabiensis (62.5%, n = 363/580), An. gambiae sensu stricto (An. gambiae s.s.; 0.7%, n = 4/580), Anopheles merus (0.7%, n = 4/580) and Anopheles quadriannulatus (0.2%, n = 1/580), with the remaining samples (35.5%, n = 206/580) unamplified. Anopheles funestus s.l. was composed of An. rivulorum (14.6%, n = 15/103) and An. leesoni (11.6%, n = 12/103); the remaining samples were unamplified (73.8%, n = 76/103). A total of 981 samples were subjected to PCR analysis for malaria parasite detection; of these 16 (1.6%) were confirmed to be positive for Plasmodium falciparum. The overall human blood index was 0.13 (32/238). CONCLUSIONS: Anopheles gambiae, An. funestus and An. coustani are key malaria vectors in the Taveta region of Kenya, showing concurrent indoor and outdoor transmission. All of the vectors tested showed a higher propensity for bovine and goat blood than for human blood.

A prospective cohort study of Plasmodium falciparum malaria in three sites of Western Kenya.

BACKGROUND: Malaria in western Kenya is currently characterized by sustained high Plasmodial transmission and infection resurgence, despite positive responses in some areas following intensified malaria control interventions since 2006. This study aimed to evaluate long-term changes in malaria transmission profiles and to assess patterns of asymptomatic malaria infections in school children aged 5-15 years at three sites in western Kenya with heterogeneous malaria transmission and simultaneous malaria control interventions. METHODS: The study was conducted from 2018 to 2019 and is based on data taken every third year from 2005 to 2014 during a longitudinal parasitological and mosquito adult surveillance and malaria control programme that was initiated in 2002 in the villages of Kombewa, Iguhu, and Marani. Plasmodium spp. infections were determined using microscopy. Mosquito samples were identified to species and host blood meal source and sporozoite infections were assayed using polymerase chain reaction. RESULTS: Plasmodium falciparum was the only malaria parasite evaluated during this study (2018-2019). Asymptomatic malaria parasite prevalence in school children decreased in all sites from 2005 to 2008. However, since 2011, parasite prevalence has resurged by > 40% in Kombewa and Marani. Malaria vector densities showed similar reductions from 2005 to 2008 in all sites, rose steadily until 2014, and decreased again. Overall, Kombewa had a higher risk of infection compared to Iguhu (χ2 = 552.52, df = 1, P < 0.0001) and Marani (χ2 = 1127.99, df = 1, P < 0.0001). There was a significant difference in probability of non-infection during malaria episodes (log-rank test, χ2 = 617.59, df = 2, P < 0.0001) in the study sites, with Kombewa having the least median time of non-infection during malaria episodes. Gender bias toward males in infection was observed (χ2 = 27.17, df = 1, P < 0.0001). The annual entomological inoculation rates were 5.12, 3.65, and 0.50 infective bites/person/year at Kombewa, Iguhu, and Marani, respectively, during 2018 to 2019. CONCLUSIONS: Malaria prevalence in western Kenya remains high and has resurged in some sites despite continuous intervention efforts. Targeting malaria interventions to those with asymptomatic infections who serve as human reservoirs might decrease malaria transmission and prevent resurgences. Longitudinal monitoring enables detection of changes in parasitological and entomological profiles and provides core baseline data for the evaluation of vector interventions and guidance for future planning of malaria control.

Urban Ecology of Arboviral Mosquito Vectors Along the Kenyan Coast.

The purpose of this study was to determine the ecology of the common arboviral mosquito vectors in Mombasa, Kilifi and Malindi urban areas of coastal Kenya. Mosquito larvae were collected using standard dippers and pipettes. Egg survivorship in dry soil was evaluated by collecting soil samples from dry potential larval developmental sites, re-hydrating them for hatching and rearing of the eventual larvae to adults. Adult mosquitoes were collected with CDC light traps and BG-Sentinel traps. All blood-fed females were tested for bloodmeal origin. Mosquitoes were screened for arboviruses using RT-qPCR. Overall, the predominant species were Culex quinquefasciatus (Say) 72.4% (n = 2,364) and Aedes aegypti (L.), 25.7%, (n = 838). A total of 415 larval developmental sites were identified indoors (n = 317) and outdoors (n = 98). The most productive larval developmental sites, both indoors and outdoors, were assorted small containers, water tanks, drainages, drums, and jerricans. Overall, 62% (n = 18) of the soil samples collected were positive for larvae which were used as a proxy to measure the presence of eggs. The mosquitoes fed on humans (29.8%) and chickens (3.7%). Of 259 mosquitoes tested for viral infection, 11.6% were positive for Flavivirus only. The most productive larval developmental sites for arboviral vectors indoors were small containers, water tanks, jerricans, and drums whereas small containers, water tanks, drainage channels, buckets, tires, and water troughs were the productive larval developmental sites outdoors.

Anopheles larval abundance and diversity in three rice agro-village complexes Mwea irrigation scheme, central Kenya.

BACKGROUND: The diversity and abundance of Anopheles larvae has significant influence on the resulting adult mosquito population and hence the dynamics of malaria transmission. Studies were conducted to examine larval habitat dynamics and ecological factors affecting survivorship of aquatic stages of malaria vectors in three agro-ecological settings in Mwea, Kenya. METHODS: Three villages were selected based on rice husbandry and water management practices. Aquatic habitats in the 3 villages representing planned rice cultivation (Mbui Njeru), unplanned rice cultivation (Kiamachiri) and non-irrigated (Murinduko) agro-ecosystems were sampled every 2 weeks to generate stage-specific estimates of mosquito larval densities, relative abundance and diversity. Records of distance to the nearest homestead, vegetation coverage, surface debris, turbidity, habitat stability, habitat type, rice growth stage, number of rice tillers and percent Azolla cover were taken for each habitat. RESULTS: Captures of early, late instars and pupae accounted for 78.2%, 10.9% and 10.8% of the total Anopheles immatures sampled (n = 29,252), respectively. There were significant differences in larval abundance between 3 agro-ecosystems. The village with 'planned' rice cultivation had relatively lower Anopheles larval densities compared to the villages where 'unplanned' or non-irrigated. Similarly, species composition and richness was higher in the two villages with either 'unplanned' or limited rice cultivation, an indication of the importance of land use patterns on diversity of larval habitat types. Rice fields and associated canals were the most productive habitat types while water pools and puddles were important for short periods during the rainy season. Multiple logistic regression analysis showed that presence of other invertebrates, percentage Azolla cover, distance to nearest homestead, depth and water turbidity were the best predictors for Anopheles mosquito larval abundance. CONCLUSION: These results suggest that agricultural practices have significant influence on mosquito species diversity and abundance and that certain habitat characteristics favor production of malaria vectors. These factors should be considered when implementing larval control strategies which should be targeted based on habitat productivity and water management.

Spatial distribution, blood feeding pattern, and role of Anopheles funestus complex in malaria transmission in central Kenya.

Studies were conducted to determine the role of sibling species of Anopheles funestus complex in malaria transmission in three agro-ecosystems in central Kenya. Mosquitoes were sampled indoors and outdoors, and rDNA PCR was successfully used to identify 340 specimens. Anopheles parensis (91.8%), A. funestus (6.8%), and Anopheles leesoni (1.5%) were the three sibling species identified. A. parensis was the dominant species at all study sites, while 22 of 23 A. funestus were collected in the non-irrigated study site. None of the 362 specimens tested was positive for Plasmodium falciparum circumsporozoite proteins by enzyme-linked immunosorbent assay. The most common blood-meal sources (mixed blood meals included) for A. parensis were goat (54.0%), human (47.6%), and bovine (39.7%), while the few A. funestus s.s. samples had fed mostly on humans. The human blood index (HBI) for A. parensis (mixed blood meals included) in the non-irrigated agro-ecosystem was 0.93 and significantly higher than 0.33 in planned rice agro-ecosystem. The few samples of A. funestus s.s. and A. funestus s.l. also showed a trend of higher HBI in the non-irrigated agro-ecosystem. We conclude that agricultural practices have significant influence on distribution and blood feeding behavior of A. funestus complex. Although none of the species was implicated with malaria transmission, these results may partly explain why non-irrigated agro-ecosystems are associated with higher risk of malaria transmission by this species compared to irrigated agro-ecosystems.

Larval habitat diversity and mosquito species distribution along the coast of Kenya.

Background: Management of arboviruses relies heavily on vector control. Implementation and sustenance of effective control measures requires regular surveillance of mosquito occurrences, species abundance and distribution. The current study evaluated larval habitat diversity and productivity, mosquito species diversity and distribution in selected sites along the coast of Kenya. Methods: A cross-sectional survey of mosquito breeding habitats, species diversity and distribution was conducted in urban, peri-urban and forested ecological zones in Mombasa and Kilifi counties. Results: A total of 13,009 immature mosquitoes were collected from 17 diverse aquatic habitats along the coast of Kenya. Larval productivity differed significantly (F (16, 243) = 3.21, P < 0.0001) among the aquatic habitats, with tyre habitats recording the highest larval population. Culex pipiens (50.17%) and Aedes aegypti (38.73%) were the dominant mosquito species in urban areas, while Ae. vittatus (89%) was the dominant species in forested areas.  In total, 4,735 adult mosquitoes belonging to 19 species were collected in Haller Park, Bamburi, Gede and Arabuko Sokoke forest. Urban areas supported higher densities of Ae. aegypti compared to peri-urban and forest areas, which, on the other hand, supported greater mosquito species diversity. Conclusions: High Ae. aegypti production in urban and peri-urban areas present a greater risk of arbovirus outbreaks. Targeting productive habitats of Aedes aegypti, such as discarded tyres, containers and poorly maintained drainage systems in urban areas and preventing human-vector contact in peri-urban and forested areas could have a significant impact on the prevalence of arboviruses along the coast of Kenya, forestalling the periodic outbreaks experienced in the region.

Effect of rice cultivation on malaria transmission in central Kenya.

A 12-month field study was conducted between April 2004 and March 2005 to determine the association between irrigated rice cultivation and malaria transmission in Mwea, Kenya. Adult mosquitoes were collected indoors twice per month in three villages representing non-irrigated, planned, and unplanned rice agro-ecosystems and screened for blood meal sources and Plasmodium falciparum circumsporozoite proteins. Anopheles arabiensis Patton and An. funestus Giles comprised 98.0% and 1.9%, respectively, of the 39,609 female anophelines collected. Other species including An. pharoensis Theobald, An. maculipalpis Giles, An. pretoriensis Theobald, An. coustani Laveran, and An. rufipes Gough comprised the remaining 0.1%. The density of An. arabiensis was highest in the planned rice village and lowest in the non-irrigated village and that of An. funestus was significantly higher in the non-irrigated village than in irrigated ones. The human blood index (HBI) for An. arabiensis was significantly higher in the non-irrigated village compared with irrigated villages. For An. funestus, the HBI for each village differed significantly from the others, being highest in the non-irrigated village and lowest in the planned rice village. The sporozoite rate and annual entomologic inoculation rate (EIR) for An. arabiensis was 1.1% and 3.0 infective bites per person, respectively with no significant difference among villages. Sporozoite positive An. funestus were detected only in planned rice and non-irrigated villages. Overall, 3.0% of An. funestus samples tested positive for Plasmodium falciparum sporozoites. The annual EIR of 2.21 for this species in the non-irrigated village was significantly higher than 0.08 for the planned rice village. We conclude that at least in Mwea Kenya, irrigated rice cultivation may reduce the risk of malaria transmission by An. funestus but has no effect on malaria transmission by An. arabiensis. The zoophilic tendency of malaria vectors in irrigated areas accounts partly for low malaria transmission rates despite the presence of higher vector densities, highlighting the potential of zooprophylaxis in malaria control.

Host choice and multiple blood feeding behaviour of malaria vectors and other anophelines in Mwea rice scheme, Kenya.

BACKGROUND: Studies were conducted between April 2004 and February 2006 to determine the blood-feeding pattern of Anopheles mosquitoes in Mwea Kenya. METHODS: Samples were collected indoors by pyrethrum spay catch and outdoors by Centers for Disease Control light traps and processed for blood meal analysis by an Enzyme-linked Immunosorbent Assay. RESULTS: A total of 3,333 blood-fed Anopheles mosquitoes representing four Anopheles species were collected and 2,796 of the samples were assayed, with Anopheles arabiensis comprising 76.2% (n = 2,542) followed in decreasing order by Anopheles coustani 8.9% (n = 297), Anopheles pharoensis 8.2% (n = 272) and Anopheles funestus 6.7% (n = 222). All mosquito species had a high preference for bovine (range 56.3-71.4%) over human (range 1.1-23.9%) or goat (0.1-2.2%) blood meals. Some individuals from all the four species were found to contain mixed blood meals. The bovine blood index (BBI) for An. arabiensis was significantly higher for populations collected indoors (71.8%), than populations collected outdoors (41.3%), but the human blood index (HBI) did not differ significantly between the two populations. In contrast, BBI for indoor collected An. funestus (51.4%) was significantly lower than for outdoor collected populations (78.0%) and the HBI was significantly higher indoors (28.7%) than outdoors (2.4%). Anthropophily of An. funestus was lowest within the rice scheme, moderate in unplanned rice agro-ecosystem, and highest within the non-irrigated agro-ecosystem. Anthropophily of An. arabiensis was significantly higher in the non-irrigated agro-ecosystem than in the other agro-ecosystems. CONCLUSION: These findings suggest that rice cultivation has an effect on host choice by Anopheles mosquitoes. The study further indicate that zooprophylaxis may be a potential strategy for malaria control, but there is need to assess how domestic animals may influence arboviruses epidemiology before adapting the strategy.

Contribution of different aquatic habitats to adult Anopheles arabiensis and Culex quinquefasciatus (Diptera: Culicidae) production in a rice agroecosystem in Mwea, Kenya.

Studies were conducted to determine the contribution of diverse larval habitats to adult Anopheles arabiensis Patton and Culex quinquefasciatus Say production in a rice land agro-ecosystem in Mwea, Kenya. Two sizes of cages were placed in different habitat types to investigate the influence of non-mosquito invertebrates on larval mortalities and the contribution of each habitat type to mosquito productivities, respectively. These emergence traps had fine netting material covers to prevent adult mosquitoes from ovipositing in the area covered by the trap and immature mosquitoes from entering the cages. The emergence of Anopheles arabiensis in seeps, tire tracks, temporary pools, and paddies was 10.53%, 17.31%, 12.50%, and 2.14%, respectively, while the corresponding values for Cx. quinquefasciatus were 16.85% in tire tracks, 8.39% in temporary pools, and 5.65% in the paddies from 0.125 m3 cages during the study. Cages measuring 1 m3 were placed in different habitat types which included paddy, swamp, marsh, ditch, pool, and seep to determine larval habitat productivity. An. arabiensis was the predominant anopheline species (98.0%, n = 232), although a few Anopheles coustani Laveran (2.0%, n = 5) emerged from the habitats. The productivity for An. arabiensis larvae was 6.0 mosquitoes per m2 for the temporary pools, 5.5 for paddy, 5.4 for marsh, 2.7 for ditch, and 0.6 for seep. The Cx. quinquefasciatus larval habitat productivity was 47.8 mosquitoes per m2 for paddies, 35.7 for ditches, 11.1 for marshes, 4.2 for seeps, 2.4 for swamps, and 1.0 for temporary pools. Pools, paddy, and marsh habitat types were the most productive larval habitats for An. arabiensis while paddy, ditch, and marsh were the most productive larval habitats for Cx. quinquefasciatus. The most common non-mosquito invertebrate composition in the cages included Dytiscidae, Notonectidae, Belostomatidae, and Ephemerellidae, and their presence negatively affected the number of emergent mosquitoes from the cages. In conclusion, freshly formed habitats are the most productive aquatic habitats, while old and more permanent habitats are the least productive due to natural regulation of mosquito immatures.

Efficacy and persistence of long-lasting microbial larvicides against malaria vectors in western Kenya highlands.

BACKGROUND: Chemical-based malaria vector control interventions are threatened by the development of insecticide resistance and changes in the behavior of the vectors, and thus require the development of alternative control methods. Bacterial-based larvicides have the potential to target both insecticide resistant and outdoor-biting mosquitoes and are safe to use in the environment. However, the currently available microbial larvicide formulations have a short duration of activity requiring frequent re-applications which increase the cost of control interventions. This study was designed to evaluate the efficacy and duration of activity of two long-lasting formulations of Bacillus thuringiensis var. israelensis (Bti) and Bacillus sphaericus (Bs) (LL3 and FourStar®) under field conditions in western Kenya highlands. METHODS: Three sites were selected for this study in the highlands of western Kenya. In each site, one hundred anopheline larval habitats were selected and assigned to one of three arms: (i) LL3; (ii) FourStar®; and (iii) untreated control larval habitats. Four types of larval habitats were surveyed: abandoned gold mines, drainage canals, fish ponds and non-fish ponds. The habitats were sampled for mosquito larvae by using a standard dipping technique and collected larvae were recorded according to the larval stages of the different Anopheles species. The larvicides were applied at manufacturers' recommended dosage of 1 briquette per 100 square feet. Both treatment and control habitats were sampled for mosquito larvae immediately before treatment (day 0), and then at 24 hours, 3 days and weekly post-treatment for 5 months. RESULTS: Overall larval density in treatment habitats was significantly reduced after application of the two microbial larvicides as compared to the control habitats. Post-intervention reduction in anopheline larval density by LL3 was 65, 71 and 84% for 1 day, 2 weeks and 4 weeks, respectively. FourStar® reduced anopheline larval density by 60, 66 and 80% for 1 day, 2 weeks and 4 weeks, respectively. Comparisons between the treatments reveal that LL3 and FourStar® were similar in efficacy. A higher reduction in Anopheles larval density was observed in the abandoned goldmines, while drainage canals had the lowest reduction. CONCLUSIONS: Both LL3 and FourStar® long-lasting microbial larvicides were effective in reducing immature stages of An. gambiae complex and An. funestus group species, with significant reductions lasting for three months post-application.

Evaluation of four sampling techniques for surveillance of Culex quinquefasciatus (Diptera: Culicidae) and other mosquitoes in African rice agroecosystems.

Field studies were conducted in a rice, Oryza sativa L., agroecosystem in Mwea Kenya to compare the efficiency of CO2-baited Centers for Disease Control (CDC) light traps against nonbaited CDC light traps and gravid traps against oviposition traps in outdoor collection of Culex quinquefasciatus Say (Diptera: Culicidae) and other mosquitoes. Collectively, 21 mosquito species from the genera Culex, Anopheles, Mansonia, Ficalbia, and Aedes were captured during the 10-wk study period. Cx. quinquefasciatus was the predominant species in all trap types with proportions ranging from 57% in the nonbaited CDC light traps to 95% in the gravid traps. Significantly higher numbers of Cx. quinquefasciatus and Culex annulioris Theobald were collected in the CO2-baited CDC light traps than in the nonbaited CDC light traps, but the numbers of other mosquito species, including malaria vectors Anopheles arabiensis Patton and Anopheles funestus Giles did not differ significantly between the two trap types. More Cx. quinquefasciatus females were collected in grass infusion-baited gravid traps than egg rafts of this species in oviposition traps containing the same infusion. Although most mosquitoes captured in CO,-baited and nonbaited CDC light traps were unfed, most of those collected in gravid traps were gravid. From these findings, it is concluded that at least in the rice-growing area of Mwea Kenya, CO2-baited CDC light traps in conjunction with gravid traps can be used in monitoring of Cx. quinquefasciatus both for control and disease surveillance.

Mosquito species succession and physicochemical factors affecting their abundance in rice fields in Mwea, Kenya.

The succession of mosquito species and abiotic factors affecting their distribution and abundance in rice (Oryza spp.) fields was investigated over a 16-wk rice growing cycle covering the period between January and May 2006. Fifteen experimental rice plots were sampled for mosquito larvae and characterized based on rice height, number of tillers, floating vegetation cover, water depth, water temperature, turbidity, salinity, pH, dissolved oxygen, total dissolved solids, and conductivity. Microscopic identification of 3,025 larvae yielded nine mosquito species predominated by Anopheles arabiensis Patton (45.0%), Culex quinquefasciatus Say (35.8%), Anopheles pharoensis Theobald (9.0%) and Ficalbia splendens Theobald (7.1%). Other species, including Anopheles rufipes Gough, Anopheles coustani Laveran, Anonopheles maculipalpis Giles, Culex annulioris Theobald, and Culex poicilipes Theobald made up 3.1% of the total collection. Anopheles gambiae s.l., Cx. quinquefasciatus, and An. pharoensis occurred throughout the cycle, but they were more abundant up to 4 wk posttransplanting with peaks after fertilizer application. As rice plants became established, three groups of mosquitoes were recognized: the first groups included An. rufipes, Fl. splendens, and Cx. annulioris, which occurred throughout much of the second half of the rice cycle, whereas the second group included Cx. poicilipes, which was found in the middle of the rice cycle. An. coustani and An. maculipalpis formed the third group occurring toward the end of the cycle. Dissolved oxygen, number of tillers, and rice height were negatively associated with the abundance ofAn. arabiensis and Cx. quinquefasciatus larvae. In addition, Cx. quinquefasciatus also was associated with water depth (-ve) and turbidity (+ve). Abundance of An. pharoensis larvae was significantly associated with water temperature (+ve), the number of tillers (-ve), and rice height (-ve), whereas Fl. splendens was significantly associated with the number of tillers (+ve). The results demonstrate a complex nature of the interactions between some of the factors in the ecosystem and mosquito species abundance and calls for time-dependent and species-specific mosquito control operations.

Remote and field level quantification of vegetation covariates for malaria mapping in three rice agro-village complexes in Central Kenya.

BACKGROUND: We examined algorithms for malaria mapping using the impact of reflectance calibration uncertainties on the accuracies of three vegetation indices (VI)'s derived from QuickBird data in three rice agro-village complexes Mwea, Kenya. We also generated inferential statistics from field sampled vegetation covariates for identifying riceland Anopheles arabiensis during the crop season. All aquatic habitats in the study sites were stratified based on levels of rice stages; flooded, land preparation, post-transplanting, tillering, flowering/maturation and post-harvest/fallow. A set of uncertainty propagation equations were designed to model the propagation of calibration uncertainties using the red channel (band 3: 0.63 to 0.69 microm) and the near infra-red (NIR) channel (band 4: 0.76 to 0.90 microm) to generate the Normalized Difference Vegetation Index (NDVI) and the Soil Adjusted Vegetation Index (SAVI). The Atmospheric Resistant Vegetation Index (ARVI) was also evaluated incorporating the QuickBird blue band (Band 1: 0.45 to 0.52 microm) to normalize atmospheric effects. In order to determine local clustering of riceland habitats Gi*(d) statistics were generated from the ground-based and remotely-sensed ecological databases. Additionally, all riceland habitats were visually examined using the spectral reflectance of vegetation land cover for identification of highly productive riceland Anopheles oviposition sites. RESULTS: The resultant VI uncertainties did not vary from surface reflectance or atmospheric conditions. Logistic regression analyses of all field sampled covariates revealed emergent vegetation was negatively associated with mosquito larvae at the three study sites. In addition, floating vegetation (-ve) was significantly associated with immature mosquitoes in Rurumi and Kiuria (-ve); while, turbidity was also important in Kiuria. All spatial models exhibit positive autocorrelation; similar numbers of log-counts tend to cluster in geographic space. The spectral reflectance from riceland habitats, examined using the remote and field stratification, revealed post-transplanting and tillering rice stages were most frequently associated with high larval abundance and distribution. CONCLUSION: NDVI, SAVI and ARVI generated from QuickBird data and field sampled vegetation covariates modeled cannot identify highly productive riceland An. arabiensis aquatic habitats. However, combining spectral reflectance of riceland habitats from QuickBird and field sampled data can develop and implement an Integrated Vector Management (IVM) program based on larval productivity.

Environmental covariates of Anopheles arabiensis in a rice agroecosystem in Mwea, Central Kenya.

Water quality of aquatic habitats is an important determinant of female mosquito oviposition and successful larval development. This study examined the influence of environmental covariates on Anopheles arabiensis mosquito abundance in the Mwea Irrigation Scheme, Central Province of Kenya, prior to implementation of a malaria vector control program. Experimental rice plots were used to examine the environmental covariates responsible for regulating abundance and diversity of the aquatic stages of malaria vectors. Mosquito larval sampling and water quality analysis were done weekly from the flooding stage to the rice maturation stage. Sampling for mosquito larvae was conducted using standard dipping technique. During each larval collection, environmental covariates such as pH, temperature, conductivity, salinity, dissolved oxygen, water depth, and rice stage were measured. Anopheles arabiensis larval density was highest between 1 wk before transplanting and 4 wk after transplanting with peaks at weeks 0, 3, and 8. The fluctuation in values of the various environmental covariates showed characteristic patterns in different rice growth phases depending on the changes taking place due to the agronomic practices. Using a backward linear regression model, the factors that were found to be associated with abundance of An. arabiensis larvae at any of the rice growing phases included the following: dissolved oxygen, pH, turbidity, water depth, rice height, number of rice tillers, salinity, conductivity, and temperature. The environmental covariates associated with abundance of An. arabiensis were associated with early vegetative stage of the rice growth. For effective control of developmental stages of mosquito larvae, the application of larvicides should be done at the vegetative stage and the larvicides should persist until the beginning of the reproductive stage of the rice.

Current status of insecticide resistance among malaria vectors in Kenya.

BACKGROUND: Insecticide resistance has emerged as one of the major challenges facing National Malaria Control Programmes in Africa. A well-coordinated national database on insecticide resistance (IRBase) can facilitate the development of effective strategies for managing insecticide resistance and sustaining the effectiveness of chemical-based vector control measures. The aim of this study was to assemble a database on the current status of insecticide resistance among malaria vectors in Kenya. METHODS: Data was obtained from published literature through PubMed, HINARI and Google Scholar searches and unpublished literature from government reports, research institutions reports and malaria control programme reports. Each data source was assigned a unique identification code and entered into Microsoft Excel 2010 datasheets. Base maps on the distribution of insecticide resistance and resistance mechanisms among malaria vectors in Kenya were generated using ArcGIS Desktop 10.1 (ESRI, Redlands, CA, USA). RESULTS: Insecticide resistance status among the major malaria vectors in Kenya was reported in all the four classes of insecticides including pyrethroids, carbamates, organochlorines and organophosphates. Resistance to pyrethroids has been detected in Anopheles gambiae (s.s.), An. arabiensis and An. funestus (s.s.) while resistance to carbamates was limited to An. gambiae (s.s.) and An. arabiensis. Resistance to the organochlorine was reported in An. gambiae (s.s.) and An. funestus (s.s.) while resistance to organophosphates was reported in An. gambiae (s.l.) only. The mechanisms of insecticide resistance among malaria vectors reported include the kdr mutations (L 1014S and L 1014F) and elevated activity in carboxylesterase, glutathione S-transferases (GST) and monooxygenases. The kdr mutations L 1014S and L 1014F were detected in An. gambiae (s.s.) and An. arabiensis populations. Elevated activity of monooxygenases has been detected in both An. arabiensis and An. gambiae (s.s.) populations while the elevated activity of carboxylesterase and GST has been detected only in An. arabiensis populations. CONCLUSIONS: The geographical maps show the distribution of insecticide resistance and resistance mechanisms among malaria vectors in Kenya. The database generated will provide a guide to intervention policies and programmes in the fight against malaria.

Survival of immature Anopheles arabiensis (Diptera: Culicidae) in aquatic habitats in Mwea rice irrigation scheme, central Kenya.

BACKGROUND: The survivorship and distribution of Anopheles arabiensis larvae and pupae was examined in a rice agro-ecosystem in Mwea Irrigation Scheme, central Kenya, from August 2005 to April 2006, prior to implementation of larval control programme. METHODS: Horizontal life tables were constructed for immatures in semi-field condition. The time spent in the various immature stages was determined and survival established. Vertical life tables were obtained from five paddies sampled by standard dipping technique. RESULTS: Pre-adult developmental time for An. arabiensis in the trays in the experimental set up in the screen house was 11.85 days from eclosion to emergence. The mean duration of each instar stage was estimated to be 1.40 days for first instars, 2.90 days for second instars, 1.85 days for third instars, 3.80 days for fourth instars and 1.90 days for pupae. A total of 590 individuals emerged into adults, giving an overall survivorship from L1 to adult emergence of 69.4%. A total of 4,956 An. arabiensis immatures were collected in 1,400 dips throughout the sampling period. Of these, 55.9% were collected during the tillering stage, 42.5% during the transplanting period and 1.6% during the land preparation stage. There was a significant difference in the An. arabiensis larval densities among the five stages. Also there was significant variation in immature stage composition for each day's collection in each paddy. These results indicate that the survival of the immatures was higher in some paddies than others. The mortality rate during the transplanting was 99.9% and at tillering was 96.6%, while the overall mortality was 98.3%. CONCLUSION: The survival of An. arabiensis immatures was better during the tillering stage of rice growth. Further the survival of immatures in rice fields is influenced by the rice agronomic activities including addition of nitrogenous fertilizers and pesticides. For effective integrated vector management, the application of larvicides should target An. arabiensis larvae at the tillering stage (early vegetative stage of rice) when their survival in the aquatic habitats is high to significantly reduce them and the larvicides should be long-lasting to have a significant impact on the malaria vector productivity on the habitats.

Spatially targeting Culex quinquefasciatus aquatic habitats on modified land cover for implementing an Integrated Vector Management (IVM) program in three villages within the Mwea Rice Scheme, Kenya.

BACKGROUND: Continuous land cover modification is an important part of spatial epidemiology because it can help identify environmental factors and Culex mosquitoes associated with arbovirus transmission and thus guide control intervention. The aim of this study was to determine whether remotely sensed data could be used to identify rice-related Culex quinquefasciatus breeding habitats in three rice-villages within the Mwea Rice Scheme, Kenya. We examined whether a land use land cover (LULC) classification based on two scenes, IKONOS at 4 m and Landsat Thematic Mapper at 30 m could be used to map different land uses and rice planted at different times (cohorts), and to infer which LULC change were correlated to high density Cx. quinquefasciatus aquatic habitats. We performed a maximum likelihood unsupervised classification in Erdas Imagine V8.7 and generated three land cover classifications, rice field, fallow and built environment. Differentially corrected global positioning systems (DGPS) ground coordinates of Cx. quinquefasciatus aquatic habitats were overlaid onto the LULC maps generated in ArcInfo 9.1. Grid cells were stratified by levels of irrigation (well-irrigated and poorly-irrigated) and varied according to size of the paddy. RESULTS: Total LULC change between 1988-2005 was 42.1 % in Kangichiri, 52.8 % in Kiuria and and 50.6 % Rurumi. The most frequent LULC changes was rice field to fallow and fallow to rice field. The proportion of aquatic habitats positive for Culex larvae in LULC change sites was 77.5% in Kangichiri, 72.9% in Kiuria and 73.7% in Rurumi. Poorly - irrigated grid cells displayed 63.3% of aquatic habitats among all LULC change sites. CONCLUSION: We demonstrate that optical remote sensing can identify rice cultivation LULC sites associated with high Culex oviposition. We argue that the regions of higher Culex abundance based on oviposition surveillance sites reflect underlying differences in abundance of larval habitats which is where limited control resources could be concentrated to reduce vector larval abundance.