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Accurate identification of disease vector insects is crucial when collecting epidemiological data. Traditionally, mosquitoes that transmit diseases like malaria, yellow fever, chikungunya, and dengue fever have been identified by looking at their external morphological features at different life cycle stages. This process is tedious and labour intensive.In this paper, the potential of Raman spectroscopy in combination with Linear and Quadratic Discriminant Analysis to classify three mosquito species, namely: Aedes aegypti, Anopheles gambiae and Culex quinquefasciatus, was explored. The classification was based on the mosquitoes’ cuticular melanin. The three mosquito species represented two subfamilies of medically important mosquitoes, i.e. the Anophelinae and the Culicinae. The housefly (Musca domestica) was included as a ‘control’ group to assess the discrimination ability of the classifiers. This study is the first to use Raman spectroscopy to classify mosquitoes. Fresh mosquitoes were anaesthetized with chloroform, and a dispersive Raman microscope was used to capture spectra from their legs. Broad melanin peaks centred around 1400 cm-1, 1590 cm-1, and 2060 cm-1 dominated the spectra. Variance Threshold (VT) and Principal Component Analysis (PCA) were used for feature selection and feature extraction respectively from the preprocessed data. The extracted features were then used to train and test Linear Discriminant Analysis (LDA) and Quadratic Discriminant Analysis (QDA) classifiers.The VT/PCA/QDA classification model performed better than VT/PCA/LDA. VT/PCA/QDA achieved an overall accuracy of 94%, sensitivity of 87% and specificity of 96%, whereas VT/PCA/LDA attained an accuracy of 85%, a sensitivity of 69% and a specificity of 90%. The success of these relatively simple classification models on Raman spectroscopy data lays the groundwork for future development of models for discriminating morphologically indistinguishable insect species.

Background: Behavioural responses of mosquito vectors to insecticide-treated materials are critical aspects of disease control and for insecticide resistance management. Physiological resistance to insecticides may change mosquitoes’ behavioural response to insecticide-treated materials such as bed nets. Methods: Using free-flying bioassay conducted in cubic-shape cages with different net materials on different sides of the cage, we examined the behavioural responses of insecticide-resistant (Bungoma strain) and susceptible (Kisumu strain) Anopheles gambiae sensu stricto mosquitoes to different types of long-lasting insecticidal nets (LLINs) including the piperonyl butoxide treated (PBO) LLIN using untreated nets as control. We conducted video tracking of mosquito flying paths and analysed landing attempts, resting preference and resting duration on treated and untreated netting materials. Kisumu susceptible strain (An. gambiae s.s) was used as a control.Results: We found that for both field and susceptible strain mosquitoes PBO nets caused significantly higher mortalities. Direct observation found that the resistant strain mosquitoes showed significant inactive overall compared to susceptible mosquitoes regardless of net material. Direct observations found that resistant mosquitoes showed 5-10 fold higher proportion of resting on the non-treated nets (10~20% resting rate) compared to treated nets (Conclusion: Based on the overall behaviour analysis, the evidence suggested that the resistant An. gambiae s.s. mosquitoes were inactive against insecticide-treated nets. The results from this study explain why mortality in resistant mosquitoes was low in the presence of LLINs – they did not contact often and did not rest often on the treated nets.

Background Accurate malaria diagnosis and appropriate treatment at local health facilities are critical to reducing morbidity and human reservoir of infectious gametocytes. The current study assessed the accuracy of malaria diagnosis and treatment practices in three health care facilities in rural western Kenya. Methods The accuracy of malaria detection and treatment recommended compliance was monitored in two public and one private hospital from November 2019 through March 2020. Blood smears from febrile patients were examined by hospital laboratory technicians and re-examined by an expert microscopists thereafter subjected to real-time polymerase chain reaction (RT-PCR) for quality assurance. In addition, blood smears from patients diagnosed with malaria rapid diagnostic tests (RDT) and presumptively treated with anti-malarial were re-examined by an expert microscopist. Results A total of 1131 febrile outpatients were assessed for slide positivity (936), RDT (126) and presumptive diagnosis (69). The overall positivity rate for Plasmodium falciparum was 28% (257/936). The odds of slide positivity was higher in public hospitals, 30% (186/624, OR:1.44, 95% CI = 1.05–1.98, p p p p = 0.004). RDT was the most sensitive (73.8%, 95% CI = 39.5–57.4) and specific (89.2%, 95% CI = 78.5–95.2) followed by hospital microscopy (sensitivity 47.6%, 95% CI = 38.2–57.1) and specificity (86.7%, 95% CI = 80.8–91.0). Presumptive diagnosis had the lowest sensitivity (25.7%, 95% CI = 13.1–43.6) and specificity (75.0%, 95% CI = 50.6–90.4). RDT had the highest non-treatment of negatives [98.3% (57/58)] while hospital microscopy had the lowest [77.3% (116/150)]. Health facilities misdiagnosis was at 27.9% (77/276). PCR confirmed 5.2% (4/23) of the 77 misdiagnosed cases as false positive and 68.5% (37/54) as false negative. Conclusions The disparity in malaria diagnosis at health facilities with many slide positives reported as negatives and high presumptive treatment of slide negative cases, necessitates augmenting microscopic with RDTs and calls for Ministry of Health strengthening supportive infrastructure to be in compliance with treatment guidelines of Test, Treat, and Track to improve malaria case management.

Background: Malaria elimination and eradication efforts can be advanced by including transmission-blocking or reducing vaccines (TBVs) alongside existing interventions. Key transmission-blocking vaccine candidates, such as Pfs230 domain one and Pfs48/45 domain 3, should be genetically stable to avoid developing ineffective vaccines due to antigenic polymorphisms. We evaluated genetic polymorphism and temporal stability of Pfs230 domain one and Pfs48/45 domain three in Plasmodium falciparum parasites from western Kenya.Methods: Dry blood spots on filter paper were collected from febrile malaria patients reporting to community health facilities in endemic areas of Homa Bay and Kisumu Counties and an epidemic-prone area of Kisii County in 2018 and 2019. Plasmodium speciation was performed using eluted DNA and real-time PCR. Amplification of the target domains of the two Pfs genes was performed on P. falciparum positive samples. We sequenced Pfs230 domain one on 156 clinical isolates and Pfs48/45 domain three on 118 clinical isolates to infer the levels of genetic variability, signatures of selection, genetic diversity indices and perform other evolutionary analyses.Results:Pfs230 domain one had low nucleotide diversity (π = 0.15 × 10–2) with slight variation per study site. Six polymorphic sites with nonsynonymous mutations and eight haplotypes were discovered. I539T was a novel variant, whereas G605S was nearing fixation. Pfs48/45 domain three had a low π (0.063 × 10–2), high conservation index, and three segregating sites, resulting in nonsynonymous mutation and four haplotypes. Some loci of Pfs230 D1 were in positive or negative linkage disequilibrium, had negative or positive selection signatures, and others (1813, 1955) and (1813, 1983) had a history of recombination. Mutated loci pairs in Pfs48/45 domain three had negative linkage disequilibrium, and some had negative and positive Tajima’s D values with no history of recombination events.Conclusion: The two transmission blocking vaccine candidates have low nucleotide diversity, a small number of zone-specific variants, high nucleotide conservation index, and high frequency of rare alleles. With the near fixation a polymorphic site and the proximity of mutated codons to antibody binding epitopes, it will be necessary to continue monitoring sequence modifications of these domains when designing TBVs that include Pfs230 and Pfs48/45 antigens.

Food insecurity, recurrent famine, and poverty threaten the health of millions of African residents. Construction of dams and rural irrigation schemes is key to solving these problems. The sub-Saharan Africa International Center of Excellence for Malaria Research addresses major knowledge gaps and challenges in Plasmodium falciparum and Plasmodium vivax malaria control and elimination in malaria-endemic areas of Kenya and Ethiopia where major investments in water resource development are taking place. This article highlights progress of the International Center of Excellence for Malaria Research in malaria vector ecology and behavior, epidemiology, and pathogenesis since its inception in 2017. Studies conducted in four field sites in Kenya and Ethiopia show that dams and irrigation increased the abundance, stability, and productivity of larval habitats, resulting in increased malaria transmission and a greater disease burden. These field studies, together with hydrological and malaria transmission modeling, enhance the ability to predict the impact of water resource development projects on vector larval ecology and malaria risks, thereby facilitating the development of optimal water and environmental management practices in the context of malaria control efforts. Intersectoral collaborations and community engagement are crucial to develop and implement cost-effective malaria control strategies that meet food security needs while controlling malaria burden in local communities.

Background The gold standard for diagnosing Plasmodium falciparum infection is microscopic examination of Giemsa-stained peripheral blood smears. The effectiveness of this procedure for infection surveillance and malaria control may be limited by a relatively high parasitaemia detection threshold. Persons with microscopically undetectable infections may go untreated, contributing to ongoing transmission to mosquito vectors. The purpose of this study was to determine the magnitude and determinants of undiagnosed submicroscopic P. falciparum infections in a rural area of western Kenya. Methods A health facility-based survey was conducted, and 367 patients seeking treatment for symptoms consistent with uncomplicated malaria in Homa Bay County were enrolled. The frequency of submicroscopic P. falciparum infection was measured by comparing the prevalence of infection based on light microscopic inspection of thick blood smears versus real-time polymerase chain reaction (RT-PCR) targeting P. falciparum 18S rRNA gene. Long-lasting insecticidal net (LLIN) use, participation in nocturnal outdoor activities, and gender were considered as potential determinants of submicroscopic infections. Results Microscopic inspection of blood smears was positive for asexual P. falciparum parasites in 14.7% (54/367) of cases. All of these samples were confirmed by RT-PCR. 35.8% (112/313) of blood smear negative cases were positive by RT-PCR, i.e., submicroscopic infection, resulting in an overall prevalence by RT-PCR alone of 45.2% compared to 14.7% for blood smear alone. Females had a higher prevalence of submicroscopic infections (35.6% or 72 out of 202 individuals, 95% CI 28.9–42.3) compared to males (24.2%, 40 of 165 individuals, 95% CI 17.6–30.8). The risk of submicroscopic infections in LLIN users was about half that of non-LLIN users (OR = 0.59). There was no difference in the prevalence of submicroscopic infections of study participants who were active in nocturnal outdoor activities versus those who were not active (OR = 0.91). Patients who participated in nocturnal outdoor activities and use LLINs while indoors had a slightly higher risk of submicroscopic infection than those who did not use LLINs (OR = 1.48). Conclusion Microscopic inspection of blood smears from persons with malaria symptoms for asexual stage P. falciparum should be supplemented by more sensitive diagnostic tests in order to reduce ongoing transmission of P. falciparum parasites to local mosquito vectors.

Background Long-lasting insecticidal nets are an effective tool in reducing malaria transmission. However, with increasing insecticide resistance little is known about how physiologically resistant malaria vectors behave around a human-occupied bed net, despite their importance in malaria transmission. We used the Mbita bednet trap to assess the host-seeking behavior of insecticide-resistant Anopheles gambiae mosquitoes under semi-field conditions. The trap incorporates a mosquito netting panel which acts as a mechanical barrier that prevents host-seeking mosquitoes from reaching the human host baiting the trap. Methods Susceptible and pyrethroid-resistant colonies of female Anopheles gambiae mosquitoes aged 3–5 days old were used in this study. The laboratory-bred mosquitoes were color-marked with fluorescent powders and released inside a semi-field environment where a human subject slept inside a bednet trap erected in a traditional African hut. The netting panel inside the trap was either untreated (control) or deltamethrin-impregnated. The mosquitoes were released outside the hut. Only female mosquitoes were used. A window exit trap was installed on the hut to catch mosquitoes exiting the hut. A prokopack aspirator was used to collect indoor and outdoor resting mosquitoes. In addition, clay pots were placed outside the hut to collect outdoor resting mosquitoes. The F1 progeny of wild-caught mosquitoes were also used in these experiments. Results The mean number of resistant mosquitoes trapped in the deltamethrin-impregnated bed net trap was higher (mean = 50.21± 3.7) compared to susceptible counterparts (mean + 22.4 ± 1.31) (OR = 1.445; P Conclusion The results show that deltamethrin-treatment of netting panels inside the bednet trap did not alter the host-seeking behavior of insecticide-resistant female An. gambiae mosquitoes. On the contrary, susceptible females exited the hut and remained outdoors when a treated net was used. However, further investigations of the behavior of resistant mosquitoes under natural conditions should be undertaken to confirm these observations and improve the current intervention which are threatened by insecticide resistance and altered vector behavior.

Background An. funestus is a major Afrotropical vector of human malaria. This study sought to investigate the larval ecology, sporozoite infection rates and blood meal sources of An. funestus in western Kenya. Methods Larval surveys were carried out in Bungoma (Highland) and Kombewa (lowland) of western Kenya. Aquatic habitats were identified, characterized, georeferenced and carefully examined for mosquito larvae and predators. Indoor resting mosquitoes were sampled using pyrethrum spray catches. Adults and larvae were morphologically and molecularly identified to species. Sporozoite infections and blood meal sources were detected using real-time PCR and ELISA respectively. Results Of the 151 aquatic habitats assessed, 62/80 (78%) in Bungoma and 58/71(82%) in Kombewa were positive for mosquito larvae. Of the 3,193 larvae sampled, An. funestus larvae constitute 38% (1224/3193). Bungoma recorded a higher number of An. funestus larvae (85%, 95%, CI, 8.722–17.15) than Kombewa (15%, 95%, CI, 1.33–3.91). Molecular identification of larvae showed that 89% (n = 80) were An. funestus. Approximately 59%, 35% and 5% of An. funestus larvae co-existed with An. gambiae s.l, Culex spp and An. coustani in the same habitats respectively. Of 1,221 An. funestus s.l adults sampled, molecular identifications revealed that An. funestus constituted 87% (n = 201) and 88% (n = 179) in Bungoma and Kombewa, respectively. The Plasmodium falciparum sporozoite rate of An. funestus in Bungoma and Kombewa was 2% (3/174) and 1% (2/157), respectively, and the human blood index of An. funestus was 84% (48/57) and 89% (39/44) and for Bungoma and Kombewa, respectively. Conclusion Man-made ponds had the highest abundance of An. funestus larvae. Multiple regression and principal component analyses identified the distance to the nearest house as the key environmental factor associated with the abundance of An. funestus larvae in aquatic habitats. This study serves as a guide for the control of An. funestus and other mosquito species to complement existing vector control strategies.

Background Everyday hundreds of people, mainly men, set out to take part in a vibrant artisanal capture fishing (ACF) industry on Lake Victoria. It is not known whether actions of artisanal fishers, in their unrelenting quest for existence, surpass ecosystems’ sustainability thresholds with potentially negative repercussions on human health with respect to malaria transmission potential. This article sought to fill this information gap. Methods This study used an ecosystem approach to find out how ACF processes facilitate the breeding of mosquitoes. The observational study adopted a cross-sectional design and was carried out on Mageta Island situated inside Lake Victoria in western Kenya. Results Of the 87 mosquito larval habitats identified 27 (31%) were created through ACF activities. The ACF-related habitats, hereafter collectively referred to as ‘fishing habitats’, included fishing boats (24), trenches (1) and fish bait mines (2). About half (48%) of Anopheles larvae were recovered from fishing habitats. The mean larval density in the fishing habitats (35.7 ± 1.15) was double that in non-fishing habitats (17.4 ± 0.539). Despite being the most common ‘non-fishing habitat’ type (N = 32), the mean number of Anopheles larvae present in rock pools (30.81 ± 10.54) was significantly less than those found inside fishing boats (N = 24; 40.08 ± 10.16). Overall, man-made habitats and those used to support livelihoods contained significantly more Anopheles larvae. Conclusions These data show that artisanal capture fishing is a key driver of malaria epidemiology on Mageta Island. This suggests that larval source management strategies in the global south should pay attention to the heterogeneity in Anopheles breeding habitats created through livelihood activities.

Reliance on broad-spectrum insecticides and chemotherapeutic agents to control hematophagous insect vectors, and their related diseases is threatened by increasing insecticide and drug resistance, respectively. Thus, development of novel, alternative, complementary and effective technologies for surveillance and control of such insects is strongly encouraged. Semiochemicals are increasingly developed for monitoring and intervention of insect crop pests, but this has not been adequately addressed for hematophagous insects of medical and veterinary importance. This review provides an insight in the application of semiochemicals for control of hematophagous insects. Here, we provide specific information regarding the isolation and identification of semiochemical compounds, optimization approaches, detection, perception and discrimination by the insect olfactory system. Navigation of insects along wind-borne odor plumes is discussed and methods of odor application in field situations are reviewed. Finally, we discuss prospects and future challenges for the application of semiochemical-based tools with emphasis on mosquitoes. The acquired knowledge can guide development of more effective components of integrated vector management, safeguard against emerging resistance of insects to existing insecticides and reduce the burden of vector-borne diseases.

Genetic strategies for large scale pest or vector control using modified insects are not yet operational in Africa, and currently rely on import of the modified strains to begin preliminary, contained studies. Early involvement of research teams from participating countries is crucial to evaluate candidate field interventions. Following the recommended phased approach for novel strategies, evaluation should begin with studies in containment facilities. Experiences to prepare facilities and build international teams for research on transgenic mosquitoes revealed some important organizing themes underlying the concept of “facilities readiness,” or the point at which studies in containment may proceed, in sub-Saharan African settings. First, “compliance” for research with novel or non-native living organisms was defined as the fulfillment of all legislative and regulatory requirements. This is not limited to regulations regarding use of transgenic organisms. Second, the concept of “colony utility” was related to the characteristics of laboratory colonies being produced so that results of studies may be validated across time, sites, and strains or technologies; so that the appropriate candidate strains are moved forward toward field studies. Third, the importance of achieving “defensible science” was recognized, including that study conclusions can be traced back to evidence, covering the concerns of various stakeholders over the long term. This, combined with good stewardship of resources and appropriate funding, covers a diverse set of criteria for declaring when “facilities readiness” has been attained. It is proposed that, despite the additional demands on time and resources, only with the balance of and rigorous achievement of each of these organizing themes can collaborative research into novel strategies in vector or pest control reliably progress past initial containment studies.

The host preferences of the anthropophilic mosquito species in the Anopheles gambiae complex (Diptera: Culicidae) are mediated by skin bacterial volatiles. However, it is not known whether these mosquitoes respond differentially to skin bacterial volatiles from non-human host species. In this study, the responses of two malaria mosquito species in the An. gambiae complex, Anopheles gambiae s.s. (hereafter, An. gambiae) and Anopheles arabiensis, with different host preferences, to volatiles released from skin bacteria were tested. Skin bacteria collected from human, cow and chicken skin significantly increased trap catches; traps containing bacteria collected from human skin caught the highest proportions of An. gambiae and An. arabiensis. Traps with bacteria of human origin caught a significantly higher proportion of An. gambiae than of An. arabiensis, whereas bacterial volatiles from the chicken attracted significantly higher numbers of An. arabiensis than of An. gambiae. Additionally, An. gambiae showed a specialized response to volatiles from four specific bacteria, whereas An. arabiensis responded equally to all species of bacteria tested. Skin bacterial volatiles may therefore play important roles in guiding mosquitoes with different host preferences. The identification of these bacterial volatiles can contribute to the development of an odour blend that attracts mosquitoes with different host preferences.

The effects of formulated essential oils of Tagetes minuta and Tithonia diversifolia on Rhipicephalus appendiculatus infesting livestock were evaluated in semi-field experiments. Forty-five zebu cattle naturally infested with ticks were randomly selected from 15 herds, three animals from each. Of the three animals within each herd, one was treated with 1 g of petroleum jelly (control), one with 1 g of essential oil of T. minuta and one with 1 g of essential oil of T. diversifolia on the inner side of ear pinna, the preferred feeding site of R. appendiculatus. Tick infestation on each treated host animal was monitored daily for 18 days by counting the number of ticks attached to the animals. Within 1–4 days post-treatment, the number of ticks on animals treated with essential oils was reduced by more than half the original population. By the 5th day post-treatment, more than 75 and 60% of R. appendiculatus and other tick species, respectively, became dislodged and dropped off. A stronger repellent effect was shown by the essential oil of T. minuta than the essential oil of T. diversifolia. Mean residual protection afforded by T. minuta was 12.5 days and for T. diversifolia 7.9 days. There was no significant difference in the effectiveness of essential oils between male and female R. appendiculatus. Both T. minuta and T. diversifolia essential oils affected several other less dominant but economically important tick species. Results suggest the potential for essential oil formulations in integrated pest management of ticks and associated tick-borne diseases among resource-limited livestock farmers in tropical Africa.

Background Long Lasting Insecticidal Nets (LLINs) and indoor residual spraying (IRS) represent powerful tools for controlling malaria vectors in sub-Saharan Africa. The success of these interventions relies on their capability to inhibit indoor feeding and resting of malaria mosquitoes. This study sought to understand the interaction of insecticide resistance with indoor and outdoor resting behavioral responses of malaria vectors from Western Kenya. Methods The status of insecticide resistance among indoor and outdoor resting anopheline mosquitoes was compared in Anopheles mosquitoes collected from Kisumu and Bungoma counties in Western Kenya. The level and intensity of resistance were measured using WHO-tube and CDC-bottle bioassays, respectively. The synergist piperonyl butoxide (PBO) was used to determine if metabolic activity (monooxygenase enzymes) explained the resistance observed. The mutations at the voltage-gated sodium channel (Vgsc) gene and Ace 1 gene were characterized using PCR methods. Microplate assays were used to measure levels of detoxification enzymes if present. Results A total of 1094 samples were discriminated within Anopheles gambiae s.l. and 289 within An. funestus s.l. In Kisian (Kisumu county), the dominant species was Anopheles arabiensis 75.2% (391/520) while in Kimaeti (Bungoma county) collections the dominant sibling species was Anopheles gambiae s.s 96.5% (554/574). The An. funestus s.l samples analysed were all An. funestus s.s from both sites. Pyrethroid resistance of An.gambiae s.l F1 progeny was observed in all sites. Lower mortality was observed against deltamethrin for the progeny of indoor resting mosquitoes compared to outdoor resting mosquitoes (Mortality rate: 37% vs 51%, P = 0.044). The intensity assays showed moderate-intensity resistance to deltamethrin in the progeny of mosquitoes collected from indoors and outdoors in both study sites. In Kisian, the frequency of vgsc-L1014S and vgsc-L1014F mutation was 0.14 and 0.19 respectively in indoor resting malaria mosquitoes while those of the outdoor resting mosquitoes were 0.12 and 0.12 respectively. The ace 1 mutation was present in higher frequency in the F1 of mosquitoes resting indoors (0.23) compared to those of mosquitoes resting outdoors (0.12). In Kimaeti, the frequencies of vgsc-L1014S and vgsc-L1014F were 0.75 and 0.05 respectively for the F1 of mosquitoes collected indoors whereas those of outdoor resting ones were 0.67 and 0.03 respectively. The ace 1 G119S mutation was present in progeny of mosquitoes from Kimaeti resting indoors (0.05) whereas it was absent in those resting outdoors. Monooxygenase activity was elevated by 1.83 folds in Kisian and by 1.33 folds in Kimaeti for mosquitoes resting indoors than those resting outdoors respectively. Conclusion The study recorded high phenotypic, metabolic and genotypic insecticide resistance in indoor resting populations of malaria vectors compared to their outdoor resting counterparts. The indication of moderate resistance intensity for the indoor resting mosquitoes is alarming as it could have an operational impact on the efficacy of the existing pyrethroid based vector control tools. The use of synergist (PBO) in LLINs may be a better alternative for widespread use in these regions recording high insecticide resistance.

Long-lasting insecticidal nets and indoor residual spraying have contributed to a decline in malaria over the last decade, but progress is threatened by the development of physiological and behavioral resistance of mosquitoes against insecticides. Acknowledging the need for alternative vector control tools, we quantified the effects of eave screening in combination with a push-pull system based on the simultaneous use of a repellent (push) and attractant-baited traps (pull). Field experiments in western Kenya showed that eave screening, whether used in combination with an attractant-baited trap or not, was highly effective in reducing house entry by malaria mosquitoes. The magnitude of the effect varied for different mosquito species and between two experiments, but the reduction in house entry was always considerable (between 61% and 99%). The use of outdoor, attractant-baited traps alone did not have a significant impact on mosquito house entry but the high number of mosquitoes trapped outdoors indicates that attractant-baited traps could be used for removal trapping, which would enhance outdoor as well as indoor protection against mosquito bites. As eave screening was effective by itself, addition of a repellent was of limited value. Nevertheless, repellents may play a role in reducing outdoor malaria transmission in the peridomestic area.

Summary Background Odour baits can attract host-seeking Anopheles mosquitoes indoors and outdoors. We assessed the effects of mass deployment of odour-baited traps on malaria transmission and disease burden. Methods We installed solar-powered odour-baited mosquito trapping systems (SMoTS) to households on Rusinga Island, Lake Victoria, western Kenya (mean population 24 879), in a stepped-wedge cluster-randomised trial. All residents in the completed health and demographic surveillance system were eligible to participate. We used the travelling salesman algorithm to assign all households to a cluster (50 or 51 geographically contiguous households); nine contiguous clusters formed a metacluster. Initially, no cluster had SMoTS (non-intervened). During the course of the intervention roll-out SMoTS were gradually installed cluster by cluster until all clusters had SMoTS installed (intervened). We generated 27 cluster randomisations, with the cluster as unit of randomisation, to establish the order to install the traps in the clusters until all had a SMoTS installed. Field workers and participants were not masked to group allocation. The primary outcome of clinical malaria was monitored through repeated household visits covering the entire population, once before roll-out (baseline) and five times throughout the 2-year roll-out. We measured clinical malaria as fever plus a positive result with a rapid diagnostic test. The SolarMal project was registered on the Dutch Trial Register (NTR 3496). Findings We enrolled 34 041 participants between April 25, 2012, and March 23, 2015, to 81 clusters and nine metaclusters. 4358 households were provided with SMoTS during roll-out between June 3, 2013, and May 16, 2015. 23 clinical malaria episodes were recorded in intervened clusters and 33 episodes in non-intervened clusters (adjusted effectiveness 40·8% [95% CI −172·8 to 87·1], p=0·5) during the roll-out. Malaria prevalence measured by rapid diagnostic test was 29·8% (95% CI 20·9–38·0) lower in SMoTS clusters (prevalence 23·7%; 1552 of 6550 people) than in non-intervened clusters (prevalence 34·5%; 2002 of 5795 people). Interpretation The unexpectedly low clinical incidence of malaria during roll-out led to an imprecise estimate of effectiveness from the clinical incidence data. The substantial effect on malaria prevalence is explained by reduction in densities of Anopheles funestus . Odour-baited traps might be an effective malaria intervention. Funding COmON Foundation.

Background The World Health Organization (WHO) recommends integrated vector management (IVM) as a strategy to improve and sustain malaria vector control. However, this approach has not been widely adopted. Objectives We comprehensively assessed experiences and findings on IVM in Kenya with a view to sharing lessons that might promote its wider application. Methods The assessment used information from a qualitative external evaluation of two malaria IVM projects implemented between 2006 and 2011 and an analysis of their accumulated entomological and malaria case data. The project sites were Malindi and Nyabondo, located in coastal and western Kenya, respectively. The assessment focused on implementation of five key elements of IVM: integration of vector control methods, evidence-based decision making, intersectoral collaboration, advocacy and social mobilization, and capacity building. Results IVM was more successfully implemented in Malindi than in Nyabondo owing to greater community participation and multistakeholder engagement. There was a significant decline in the proportion of malaria cases among children admitted to Malindi Hospital, from 23.7% in 2006 to 10.47% in 2011 (p < 0.001). However, the projects’ operational research methodology did not allow statistical attribution of the decline in malaria and malaria vectors to specific IVM interventions or other factors. Conclusions Sustaining IVM is likely to require strong participation and support from multiple actors, including community-based groups, non-governmental organizations, international and national research institutes, and various government ministries. A cluster-randomized controlled trial would be essential to quantify the effectiveness and impact of specific IVM interventions, alone or in combination. Citation Mutero CM, Mbogo C, Mwangangi J, Imbahale S, Kibe L, Orindi B, Girma M, Njui A, Lwande W, Affognon H, Gichuki C, Mukabana WR. 2015. An assessment of participatory integrated vector management for malaria control in Kenya. Environ Health Perspect 123:1145–1151; http://dx.doi.org/10.1289/ehp.1408748

Behavioral responses of the malaria mosquito Anopheles coluzzii (An. gambiae sensu stricto molecular 'M form') to an expanded blend of human-derived volatiles were assessed in a dual-port olfactometer. A previously documented attractive three-component blend consisting of NH3, (S)-lactic acid, and tetradecanoic acid served as the basis for expansion. Adding 4.5 % CO2 to the basic blend significantly enhanced its attractiveness. Expansion of the blend with four human-derived C4-volatiles was then assessed, both with and without CO2. Only when CO2 was offered simultaneously, did addition of a specific concentration of 3-methyl-1-butanol or 3-methyl-butanoic acid significantly enhance attraction. The functional group at the terminal C of the 3-methyl-substituted C4 compounds influenced behavioral effectiveness. In the absence of CO2, addition of three concentrations of butan-1-amine caused inhibition when added to the basic blend. In contrast, when CO2 was added, butan-1-amine added to the basic blend strongly enhanced attraction at all five concentrations tested, the lowest being 100,000 times diluted. The reversal of inhibition to attraction by adding CO2 is unique in the class Insecta. We subsequently augmented the three-component basic blend by adding both butan-1-amine and 3-methyl-1-butanol and optimizing their concentrations in the presence of CO2 in order to significantly enhance the attractiveness to An. coluzzii compared to the three- and four-component blends. This novel blend holds potential to enhance malaria vector control based on behavioral disruption.

Background The strategy for malaria vector control in the context of reducing malaria morbidity and mortality has been the scale-up of long-lasting insecticidal nets to universal coverage and indoor residual spraying. This has led to significant decline in malaria transmission. However, these vector control strategies rely on insecticides which are threatened by insecticide resistance. In this study the status of pyrethroid resistance in malaria vectors and it’s implication in malaria transmission at the Kenyan Coast was investigated. Results Using World Health Organization diagnostic bioassay, levels of phenotypic resistance to permethrin and deltamethrin was determined. Anopheles arabiensis showed high resistance to pyrethroids while Anopheles gambiae sensu stricto (s.s.) and Anopheles funestus showed low resistance and susceptibility, respectively. Anopheles gambiae sensu lato (s.l.) mosquitoes were further genotyped for L1014S and L1014F kdr mutation by real time PCR. An allele frequency of 1.33% for L1014S with no L1014F was detected. To evaluate the implication of pyrethroid resistance on malaria transmission, Plasmodium falciparum infection rates in field collected adult mosquitoes was determined using enzyme linked immunosorbent assay and further, the behaviour of the vectors was assessed by comparing indoor and outdoor proportions of mosquitoes collected. Sporozoite infection rate was observed at 4.94 and 2.60% in An. funestus s.l. and An. gambiae s.l., respectively. A higher density of malaria vectors was collected outdoor and this also corresponded with high Plasmodium infection rates outdoor. Conclusions This study showed phenotypic resistance to pyrethroids and low frequency of L1014S kdr mutation in An. gambiae s.l. The occurrence of phenotypic resistance with low levels of kdr frequencies highlights the need to investigate other mechanisms of resistance. Despite being susceptible to pyrethroids An. funestus s.l. could be driving malaria infections in the area.

Background Most odour baits designed to attract host-seeking mosquitoes contain carbon dioxide (CO2), which enhances trap catches, given its role as a mosquito flight activator. However, the use of CO2 is expensive and logistically demanding for prolonged area-wide use. Methods This study explored the possibility of replacing organically-produced CO2 with 2-butanone in odour blends targeting host-seeking malaria mosquitoes. During semi-field and field experiments MM-X traps were baited with a human odour mimic (MB5 blend) plus CO2 or 2-butanone at varying concentrations. Unbaited traps formed a control. The attraction of Anopheles gambiae s.s., Anopheles arabiensis and Anopheles funestus to these differently baited traps was measured and mean catch sizes were compared to determine whether 2-butanone could form a viable replacement for CO2 for these target species. Results Under semi-field conditions significantly more female An. gambiae mosquitoes were attracted to a reference attractant blend (MB5 + CO2) compared to MB5 without CO2 (P

Malaria control in Africa mainly explores top-down Government-led initiatives (vertical) rather than horizontal approaches, which normally embrace active participation of communities. African malaria mosquitoes mainly breed in man-made habitats such as brick-making pits, fishponds, irrigation channels etc. This underscores the need to have communities living in affected areas to understand their role in propagating malaria and henceforth, how to contribute in its control. Malaria disproportionately affects poor people whose need for income to support basic survival far exceeds other needs. It is therefore important to integrate income generation activities (IGA) into disease control interventions. A cross-sectional survey was conducted using a questionnaire with open and closed ended questions to determine the potential integration of fish farming and mosquito control on Nyabondo plateau in western Kenya. Some of the questions asked included reasons for fish pond construction, pond condition (whether well maintained in productive state or abandoned), pond ownership (self or group), challenges faced and the respondentA¯Â?½s biodata. A total of 115 fish ponds were visited during the survey. Seventy percent of these were self owned while 30% were owned by local groups. Ponds were either maintained in active productive state or abandoned depending on the education level of the owner. Abandoned fish ponds harbored more Anopheles (malaria) mosquito larvae than active ones. Ninety nine percent of the pond owners practiced fish farming solely for income generation. There were no observable indicators that active fish farming was integrated with mosquito control. There is need to create awareness among the local communities about the importance of deliberately directing fish farming practices for integrated vector management. This will ensure proper maintenance of the ponds, assure nutrition and improve the socio-economic status of malaria burdened rural communities.

Ecological changes associated with anthropogenic ecosystem disturbances can influence human risk of exposure to malaria and other vector-borne infectious diseases. This study in Mwea, Kenya, investigated the pattern of insecticide use in irrigated and nonirrigated agroecosystems and association with the density, survival, and blood-feeding behavior of the malaria vector Anopheles arabiensis. The parity rates of adult An. arabiensis from randomly selected houses were determined by examining their ovaries for tracheal distension, and polymerase chain reaction was used to identify the host blood meals. In addition, structured questionnaires were used to generate data on insecticide use. Anopheles arabiensis densities were highest in irrigated rice agroecosystems, intermediate in irrigated French beans agroecosystems, and lowest in the nonirrigated agroecosystem. Anopheles arabiensis adult survivorship was significantly lower in irrigated rice agroecosystems than in irrigated French beans agroecosystems. The human blood index (HBI) was significantly higher in the nonirrigated agroecosystem compared to irrigated agroecosystems. Moreover, there was marked variation in HBI among villages in irrigated agroecosystems with significantly lower HBI in Kangichiri and Mathangauta compared to Kiuria, Karima, and Kangai. The proportion of mosquitoes with mixed blood meals varied among villages ranging from 0.25 in Kangichiri to 0.83 in Kiuria. Sumithion, dimethoate, and alpha cypermethrin were the most commonly used insecticides. The 1st was used mostly in irrigated rice agroecosystems, and the last 2 were used mostly in irrigated French beans agroecosystems. These findings indicate that agricultural practices may influence the ecology and behavior of malaria vectors and ultimately the risk of malaria transmission.

The health and demographic surveillance system on Rusinga Island, Western Kenya, was initiated in 2012 to facilitate a malaria intervention trial: the SolarMal project. The project aims to eliminate malaria from Rusinga Island using the nationwide adopted strategy for malaria control (insecticide-treated bed nets and case management) augmented with mass trapping of anopheline mosquitoes. The main purpose of the health and demographic surveillance is to measure the effectiveness of the trial on clinical malaria incidence, and to monitor demographic, environmental and malaria-related data variables. At the end of 2014, the 44 km(2) island had a population of approximately 25 000 individuals living in 8746 residential structures. Three times per year, all individuals are followed up and surveyed for clinical malaria. Following each round of surveillance, a randomly selected cross-section of the population is subject to a rapid diagnostic test to measure malaria. Additionally, extensive monitoring of malaria vectors is performed. Data collection and management are conducted using the OpenHDS platform, with tablet computers and applications with advanced software connected to a centralized database. Besides the general demographic information, other health-related data are collected which can be used to facilitate a range of other studies within and outside the current project. Access to the core dataset can be obtained on request from the authors.

Background Increasing levels of insecticide resistance as well as outdoor, residual transmission of malaria threaten the efficacy of existing vector control tools used against malaria mosquitoes. The development of odour-baited mosquito traps has led to the possibility of controlling malaria through mass trapping of malaria vectors. Through daily removal trapping against a background of continued bed net use it is anticipated that vector populations could be suppressed to a level where continued transmission of malaria will no longer be possible. Methods/design A stepped wedge cluster-randomised trial design was used for the implementation of mass mosquito trapping on Rusinga Island, western Kenya (the SolarMal project). Over the course of 2 years (2013–2015) all households on the island were provided with a solar-powered mosquito trapping system. A continuous health and demographic surveillance system combined with parasitological surveys three times a year, successive rounds of mosquito monitoring and regular sociological studies allowed measurement of intervention outcomes before, during and at completion of the rollout of traps. Data collection continued after achieving mass coverage with traps in order to estimate the longer term effectiveness of this novel intervention. Solar energy was exploited to provide electric light and mobile phone charging for each household, and the impacts of these immediate tangible benefits upon acceptability of and adherence to the use of the intervention are being measured. Discussion This study will be the first to evaluate whether the principle of solar-powered mass mosquito trapping could be an effective tool for elimination of malaria. If proven to be effective, this novel approach to malaria control would be a valuable addition to the existing strategies of long-lasting insecticide-treated nets and case management. Sociological studies provide a knowledge base for understanding the usage of this novel tool. Trial registration Trialregister.nl: NTR3496 – SolarMal. Registered on 20 June 2012. Electronic supplementary material The online version of this article (doi:10.1186/s13063-016-1469-z) contains supplementary material, which is available to authorized users.

Background: Large reductions in malaria transmission and mortality have been achieved over the last decade, and this has mainly been attributed to the scale-up of long-lasting insecticidal bed nets and indoor residual spraying with insecticides. Despite these gains considerable residual, spatially heterogeneous, transmission remains. To reduce transmission in these foci, researchers need to consider the local demographical, environmental and social context, and design an appropriate set of interventions. Exploring spatially variable risk factors for malaria can give insight into which human and environmental characteristics play important roles in sustaining malaria transmission. Methods: On Rusinga Island, western Kenya, malaria infection was tested by rapid diagnostic tests during two cross-sectional surveys conducted 3 months apart in 3632 individuals from 790 households. For all households demographic data were collected by means of questionnaires. Environmental variables were derived using Quickbird satellite images. Analyses were performed on 81 project clusters constructed by a traveling salesman algorithm, each containing 50-51 households. A standard linear regression model was fitted containing multiple variables to determine how much of the spatial variation in malaria prevalence could be explained by the demographic and environmental data. Subsequently, a geographically-weighted regression (GWR) was performed assuming non-stationarity of risk factors. Special attention was taken to investigate the effect of residual spatial autocorrelation and local multicollinearity. Results: Combining the data from both surveys, overall malaria prevalence was 24 %. Scan statistics revealed two clusters which had significantly elevated numbers of malaria cases compared to the background prevalence across the rest of the study area. A multivariable linear model including environmental and household factors revealed that higher socioeconomic status, outdoor occupation and population density were associated with increased malaria risk. The local GWR model improved the model fit considerably and the relationship of malaria with risk factors was found to vary spatially over the island; in different areas of the island socio-economic status, outdoor occupation and population density were found to be positively or negatively associated with malaria prevalence. Discussion: Identification of risk factors for malaria that vary geographically can provide insight into the local epidemiology of malaria. Examining spatially variable relationships can be a helpful tool in exploring which set of targeted interventions could locally be implemented. Supplementary malaria control may be directed at areas, which are identified as at risk. For instance, areas with many people that work outdoors at night may need more focus in terms of vector control. Trial registration: Trialregister.nl NTR3496 - SolarMal, registered on 20 June 2012.

Estimating the biting fraction of mosquitoes is of critical importance for risk assessment of malaria transmission. Here, we present a novel odor-based tool that has been rigorously assessed in semi-field assays and traditional African villages for estimating the number of mosquitoes that enter houses in search of a blood meal. A standard synthetic blend (SB) consisting of ammonia, (S)-lactic acid, tetradecanoic acid, and carbon dioxide was complemented with isovaleric acid, 4,5 dimethylthiazole, 2-methyl-1-butanol, and 3-methyl-1-butanol in various combinations and concentrations, and tested for attractiveness to the malaria mosquito Anopheles gambiae. Compounds were released through low density polyethylene (LDPE) material or from nylon strips (nylon). Studies were done in a semi-field facility and two traditional villages in western Kenya. The alcohol 3-methyl-1-butanol significantly increased the attraction of SB. The other compounds proved less effective or inhibitory. Tested in a village, 3-methyl-1-butanol, released from LDPE, increased the attraction of SB. Further studies showed a significantly enhanced attraction of adding 3-methyl-1-butanol to SB compared to previously-published attractive blends both under semi-field and village conditions. Other mosquito species with relevance for public health were collected with this blend in significantly higher numbers as well. These results demonstrate the advent of a novel, reliable odor-based sampling tool for the collection of malaria and other mosquitoes. The advantage of this odor-based tool over existing mosquito sampling tools is its reproducibility, objectiveness, and relatively low cost compared to current standards of CDC light traps or the human landing catch.

Host seeking by the malaria mosquito Anopheles gambiae Giles sensu stricto (Diptera: Culicidae) is mainly guided by volatile chemicals present in human odours. The skin microbiota plays an important role in the production of these volatiles, and skin bacteria grown on agar plates attract An. gambiae s.s. in the laboratory. In this study, the attractiveness of volatiles produced by human skin bacteria to An. gambiae s.s. was tested in laboratory, semi-field, and field experiments to assess these effects in increasing environmental complexity. A synthetic blend of 10 compounds identified in the headspace of skin bacteria was also tested for its attractiveness. Carbon dioxide significantly increased mosquito catches of traps baited with microbial volatiles in the semi-field experiments and was therefore added to the field traps. Traps baited with skin bacteria caught significantly more An. gambiae s.s. than control traps, both in the laboratory and semi-field experiments. Traps baited with the synthetic blend caught more mosquitoes than control traps in the laboratory experiments, but not in the semi-field experiments. Although bacterial volatiles increased mosquito catches in the field study, trapping several mosquito vector species, these effects were not significant for An. gambiae s.l. It is concluded that volatiles from skin bacteria affect mosquito behaviour under laboratory and semi-field conditions and, after fine tuning, have the potential to be developed as odour baits for mosquitoes.

Genetic modification (GM) of mosquitoes (which renders them genetically modified organisms, GMOs) offers opportunities for controlling malaria. Transgenic strains of mosquitoes have been developed and evaluation of these to 1) replace or suppress wild vector populations and 2) reduce transmission and deliver public health gains are an imminent prospect. The transition of this approach from confined laboratory settings to open field trials in disease-endemic countries (DECs) is a staged process that aims to maximize the likelihood of epidemiologic benefits while minimizing potential pitfalls during implementation. Unlike conventional approaches to vector control, application of GM mosquitoes will face contrasting expectations of multiple stakeholders, the management of which will prove critical to safeguard support and avoid antagonism, so that potential public health benefits can be fully evaluated. Inclusion of key stakeholders in decision-making processes, transfer of problem-ownership to DECs, and increased support from the wider malaria research community are important prerequisites for this. It is argued that the many developments in this field require coordination by an international entity to serve as a guiding coalition to stimulate collaborative research and facilitate stakeholder involvement. Contemporary developments in the field of modern biotechnology, and in particular GM, requires competencies beyond the field of biology, and the future of transgenic mosquitoes will hinge on the ability to govern the process of their introduction in societies in which perceived risks may outweigh rational and responsible involvement.

Malaria continues to place a disease burden on millions of people throughout the tropics, especially in sub-Saharan Africa. Although efforts to control mosquito populations and reduce human-vector contact, such as long-lasting insecticidal nets and indoor residual spraying, have led to significant decreases in malaria incidence, further progress is now threatened by the widespread development of physiological and behavioural insecticide-resistance as well as changes in the composition of vector populations. A mosquito-directed push-pull system based on the simultaneous use of attractive and repellent volatiles offers a complementary tool to existing vector-control methods. In this study, the combination of a trap baited with a five-compound attractant and a strip of net-fabric impregnated with micro-encapsulated repellent and placed in the eaves of houses, was tested in a malaria-endemic village in western Kenya. Using the repellent delta-undecalactone, mosquito house entry was reduced by more than 50%, while the traps caught high numbers of outdoor flying mosquitoes. Model simulations predict that, assuming area-wide coverage, the addition of such a push-pull system to existing prevention efforts will result in up to 20-fold reductions in the entomological inoculation rate. Reductions of such magnitude are also predicted when mosquitoes exhibit a high resistance against insecticides. We conclude that a push-pull system based on non-toxic volatiles provides an important addition to existing strategies for malaria prevention.

Background The anthropophilic malaria mosquito Anopheles gambiae sensu stricto (hereafter termed Anopheles gambiae) primarily takes blood meals from humans, whereas its close sibling Anopheles arabiensis is more opportunistic. Previous studies have identified several compounds that play a critical role in the odour-mediated behaviour of An. gambiae. This study determined the effect of natural and synthetic odour blends on mosquitoes with different host preferences to better understand the host-seeking behaviour of mosquitoes and the potential of synthetic odour blends for standardized monitoring. Methods Odour blends were initially tested for their attractiveness to An. gambiae and An. arabiensis in a semi-field system with MM-X traps baited with natural and synthetic odours. Natural host odours were collected from humans, cows and chickens. The synthetic odour blends consisted of three or five previously identified compounds released with carbon dioxide. These studies were continued under natural conditions where odour blends were tested outdoors to determine their effect on species with different host preferences. Results In the semi-field experiments, human odour attracted significantly higher numbers of both mosquito species. However, An. arabiensis was also attracted to cow and chicken odours, which confirms its opportunistic behaviour. A five-component synthetic blend was highly attractive to both mosquito species. In the field, the synthetic odour blend caught significantly more An. funestus than traps baited with human odour, while no difference was found for An. arabiensis. Catches of An. arabiensis and Culex spp. contained large numbers of blood-fed mosquitoes, mostly from cows, which indicates that these mosquitoes had fed outdoors. Conclusions Different odour baits elicit varying responses among mosquito species. Synthetic odour blends are highly effective for trapping mosquitoes; however, not all mosquitoes respond equally to the same odour blend. Combining fermenting molasses with synthetic blends in a trap represents the most effective tool to catch blood-fed mosquitoes outside houses, which is essential for understanding outdoor malaria transmission. Electronic supplementary material The online version of this article (doi:10.1186/s12936-015-0635-1) contains supplementary material, which is available to authorized users.

The use of odor baits for surveillance and control of malaria mosquitoes requires robust dispensing tools. In this study, the residual activity of a synthetic mosquito attractant blend dispensed from nylon or low density polyethylene (LDPE) sachets was evaluated at weekly intervals for one year without re-impregnation. The potential role of bacteria in modulating the attraction of mosquitoes to odor-treated nylon that had been used repeatedly over the one year study period, without re-impregnation, was also investigated. Significantly higher proportions of female Anopheles gambiae sensu stricto mosquitoes were consistently attracted to treated nylon strips than the other treatments, up to one year post-treatment. Additional volatile organic compounds and various bacterial populations were found on the treated nylon strips after one year of repeated use. The most abundant bacteria were Bacillus thuringiensis and Acinetobacter baumannii. Autoclaving of treated nylon strips prior to exposure had no effect on trap collections of laboratory-reared female An. Gambiae (P = 0.17) or wild female An. Gambiae sensu lato (P = 0.26) and Mansonia spp. (P = 0.17) mosquitoes. Trap catches of wild female An. Funestus (P < 0.001) and other anophelines (P < 0.007) were higher when treated strips had been autoclaved prior to deployment as opposed to when the treated nylon strips were not autoclaved. By contrast, wild female Culex mosquitoes were more strongly attracted to non-autoclaved compared to autoclaved treated nylon strips (P < 0.042). This study demonstrates the feasibility of using odor baits for sampling and surveillance of malaria as well as other mosquito vectors over prolonged periods of time. Preliminary evidence points towards the potential role of bacteria in sustaining prolonged use of nylon material for dispensing synthetic attractant odorants for host-seeking malaria and other mosquito vectors but further investigations are required.

Sampling indoor resting African malaria vectors is traditionally done by hand catches with oral or mechanical aspirators and pyrethrum spray catches (PSCs). In this study, we designed and briefly evaluated an inexpensive but practical alternative by using a cloth resting box or wicker resting basket and a ceiling net. Evaluations were performed in greenhouse and field situations in rural Kenya by comparing capture rates of Anopheles gambiae s.l. and Anopheles funestus (Giles) in these traps to hand collections and PSCs. A resting box and a ceiling net when used together collected more mosquitoes than a single collector using a hand-held aspirator but only one-third the number collected by PSCs. At sites where PSCs are impractical, a resting box and ceiling net can be effectively used as an alternative to hand catches in malaria surveillance.

Background There has been increasing effort in recent years to incorporate user needs in technology design and re-design. This project employed a bottom-up approach that engaged end users from the outset. Bottom-up approaches have the potential to bolster novel interventions and move them towards adaptive and evidence-based strategies. The present study concerns an innovative use of solar-powered mosquito trapping systems (SMoTS) to control malaria in western Kenya. Our paper highlights the co-dependence of research associated with the development of the SMoTS technology on one hand and research for enhancing the sustainable uptake of that very same intervention within the community on the other. Methods During the pre-intervention year, we examined the design, re-design and piloting of a novel technology to generate lessons for malaria elimination on Rusinga Island. Initial ideas about many technological necessities were evaluated and re-designed following feedback from various sources, including technical and social research as well as broader interactions with the social environment. We documented the interlocking of the multiple processes and activities that took place through process observation and document reviews. We analysed the data within the conceptual framework of system innovation by identifying mutual shaping between technical and social factors. Results Our findings illustrate how various project stakeholders including project staff, collaborators, donor, and community members simultaneously pursued interdependent technological transformations and social interests. In the ongoing process, we observed how partial outcomes in the technological domain influenced social events at a later phase and vice versa. Conclusions Looking at malaria intervention projects employing novel technologies as niches that may evolve towards system innovation, helps to reveal interrelations between the various technical and social aspects. Revealing these interrelations requires a different role for research and different perspective on innovation where innovation is more than the technical aspects. This approach therefore requires that research is designed in a way that enables obtaining feedback from both aspects. Keywords: Malaria; Co-evolution; Socio-technical; System innovation; Solar; Mosquito trap; Feedback; Community; Kenya

Background: The full-scale impact of odour-baited technology on the surveillance, sampling and control of vectors of infectious diseases is partly limited by the lack of methods for the efficient and sustainable dispensing of attractants. In this study we investigated whether locally-available and commonly used textiles are efficient substrates for the release of synthetic odorant blends attracting malaria mosquitoes. Methods: The relative efficacy of (a) polyester, (b) cotton, (c) cellulose + polyacrylate, and (d) nylon textiles as substrates for dispensing a synthetic odour blend (Ifakara blend 1(IB1)) that attracts malaria mosquitoes was evaluated in western Kenya. The study was conducted through completely randomized Latin square experimental designs under semi-field and field conditions. Results: Traps charged with IB1-impregnated polyester, cotton and cellulose + polyacrylate materials caught significantly more female Anopheles gambiae sensu stricto (semi-field conditions) and An. gambiae sensu lato (field conditions) mosquitoes than IB1-treated nylon (P = 0.001). The IB1-impregnated cellulose + polyacrylate material was the most attractive to female An. funestus mosquitoes compared to all other dispensing textile substrates (P < 0.001). The responses of female An. funestus mosquitoes to IB1-treated cotton and polyester were equal (P = 0.45). Significantly more female Culex mosquitoes were attracted to IB1-treated cotton than to the other treatments (P < 0.001). Whereas IB1-impregnated cotton and cellulose + polyacrylate material attracted equal numbers of female Mansonia mosquitoes (P = 0.44), the catches due to these two substrates were significantly higher than those associated with the other substrates (P < 0.001). Conclusion: The number and species of mosquitoes attracted to a synthetic odour blend is influenced by the type of odour-dispensing material used. Thus, surveillance and intervention programmes for malaria and other mosquito vectors using attractive odour baits should select an odour-release material that optimizes the odour blend.

Essential oils of eight plants, selected after an ethnobotanical survey conducted in Bukusu community in Bungoma County, western Kenya (Tagetes minuta, Tithonia diversifolia, Juniperus procera, Solanecio mannii, Senna didymobotrya, Lantana camara, Securidaca longepedunculata, andHoslundia opposita), were initially screened (at two doses) for their repellence against brown ear tick,Rhipicephalus appendiculatus, using a dual-choice climbing assay. The oils ofT. minutaandT. diversifoliawere then selected for more detailed study. Dose-response evaluations of these oils showed thatT. minutaoil was more repellent (RD50= 0.0021 mg) than that ofT. diversifolia(RD50= 0.263 mg). Gas chromatography-linked mass spectrometric (GC-MS) analyses showed different compositions of the two oils.T. minutaoil is comprised mainly ofcis-ocimene (43.78%), dihydrotagetone (16.71%), piperitenone (10.15%),trans-tagetone (8.67%), 3,9-epoxy-p-mentha-1,8(10)diene (6.47%),β-ocimene (3.25%), andcis-tagetone (1.95%), whereasT. diversifoliaoil is comprised mainly ofα-pinene (63.64%),β-pinene (15.00%), isocaryophyllene (7.62%), nerolidol (3.70%), 1-tridecanol (1.75%), limonene (1.52%), and sabinene (1.00%). The results provide scientific rationale for traditional use of raw products of these plants in controlling livestock ticks by the Bukusu community and lay down some groundwork for exploiting partially refined products such as essential oils of these plants in protecting cattle against infestations withR. appendiculatus.

Background Prior to the commencement of a large-scale malaria intervention study on Rusinga Island, western Kenya, intensive baseline surveillance of the mosquito population was performed using odour-baited traps. The survey aimed to determine the relative abundance and species composition of malaria vectors, and to measure seasonal and spatial heterogeneity in populations. Human malaria prevalence was combined with entomological data to provide information about malaria transmission risk before the intervention began. Materials and methods From September 2012 until June 2013, mosquito monitoring took place over successive six-week sampling periods. MM-X traps baited with attractant lures and carbon dioxide were used to collect mosquitoes from inside and outside houses, and a new random sample of 80 households was drawn for each sampling round. During the baseline period, malaria prevalence was measured twice in a randomly selected 10% of the human population. A QuickBird satellite image and digital elevation map were used to describe environmental features of the island. Mosquitoes were initially identified on the basis of morphology and anophelines were processed by PCR to confirm species identifications. Results Odour-baited MM-X traps proved to be a good tool for monitoring malaria vectors inside and outside houses. Using this tool a marked temporal and spatial heterogeneity was described for the malaria vector species Anopheles gambiae s.s., An. arabiensis and An. funestus. Regions of potentially high malaria transmission intensity were identified after mapping the distributions of malaria mosquitoes and Plasmodium-positive persons. Despite studying a range of environmental and topographical features, no strong associations were found between environmental variables and the presence or absence of adult Anopheles. Conclusions Malaria vectors and malaria prevalence are not homogeneously distributed across Rusinga Island; the risk of malaria transmission is therefore greater in some areas than others. The finding that environmental features were not closely associated with adult malaria vector distribution, indicates that other factors, such as house construction or the presence of livestock, may play a more important role in the decision of a female anopheline to approach the domestic environment of a particular house in search of a blood meal. The findings of this study demonstrate how trans-disciplinary data can be integrated to provide a better understanding of mosquito population dynamics and malaria transmission risk. Intensive mosquito monitoring before the commencement of, as well as during, a large-scale malaria intervention study, contributes valuable information which will be used in describing the eventual impact of the intervention.

Background. Most odour baits for haematophagous arthropods contain carbon dioxide (CO2). The CO2 is sourced artificially from the fermentation of refined sugar (sucrose), dry ice, pressurized gas cylinders or propane. These sources of CO2 are neither cost-effective nor sustainable for use in remote areas of sub-Saharan Africa. In this study, molasses was evaluated as a potential substrate for producing CO2 used as bait for malaria mosquitoes. Methods. The attraction of laboratory-reared and wild Anopheles gambiae complex mosquitoes to CO2 generated from yeast-fermentation of molasses was assessed under semi-field and field conditions in western Kenya. In the field, responses of wild Anopheles funestus were also assessed. Attraction of the mosquitoes to a synthetic mosquito attractant, Mbita blend (comprising ammonia, L-lactic acid, tetradecanoic acid and 3-methyl-1-butanol) when augmented with CO2 generated from yeast fermentation of either molasses or sucrose was also investigated. Results. In semi-field, the release rate of CO2 and proportion of An. gambiae mosquitoes attracted increased in tandem with an increase in the quantity of yeast-fermented molasses up to an optimal ratio of molasses and dry yeast. More An. gambiae mosquitoes were attracted to a combination of the Mbita blend plus CO2 produced from fermenting molasses than the Mbita blend plus CO2 from yeast-fermented sucrose. In the field, significantly more female An. gambiae sensu lato mosquitoes were attracted to the Mbita blend augmented with CO2 produced by fermenting 500 g of molasses compared to 250 g of sucrose or 250 g of molasses. Similarly, significantly more An. funestus, Culex and other anopheline mosquito species were attracted to the Mbita blend augmented with CO2 produced from fermenting molasses than the Mbita blend with CO2 produced from sucrose. Augmenting the Mbita blend with CO2 produced from molasses was associated with high catches of blood-fed An. gambiae s.l. and An. funestus mosquitoes. Conclusion. Molasses is a suitable ingredient for the replacement of sucrose as a substrate for the production of CO2 for sampling of African malaria vectors and other mosquito species. The finding of blood-fed malaria vectors in traps baited with the Mbita blend and CO2 derived from molasses provides a unique opportunity for the study of host-vector interactions.

Background & objectives: All organisms that are cultured for laboratory bioassays need food for sustenance and amplification of the colony to such numbers that would not compromise the progress of the research at hand. For effective turnover rate in the population generation, the diet should be such that it is readily available and provide adequate nutrients to the organisms. The aim of this study was to test and evaluate the efficacy of Rastrineobola argentea as a diet source for rearing Anopheles arabiensis Patton mosquitoes for research. Methods: Crushed fish, Rastrineobola argentea and two other commercial larval feeds (Tetramin  Baby fish food and baker's yeast) were compared for their efficacy as diet sources for feeding Anopheles arabiensis mosquito larvae. The effects of these diet types were determined against fecundity, size, longevity and male mating competitiveness.