Testing configurations of attractive toxic sugar bait (ATSB) stations in Mali, West Africa, for improving the control of malaria parasite transmission by vector mosquitoes and minimizing their effect on non-target insects.

BACKGROUND: Application methods of |Attractive Toxic Sugar Baits (ATSB) need to be improved for wide-scale use, and effects on non-target organisms (NTOs) must be assessed. The goals of this study were to determine, at the village level, the effect of different configurations of bait stations to (1) achieve < 25% Anopheles mosquito vector daily feeding rate for both males and females and (2) minimize the effect on non-target organisms. METHODS: Dye was added to Attractive Sugar Bait Stations (without toxin) to mark mosquitoes feeding on the baits, and CDC UV light traps were used to monitor for marked mosquitoes. An array of different traps were used to catch dye marked NTOs, indicating feeding on the ASB. Stations were hung on homes (1, 2, or 3 per home to optimize density) at different heights (1.0 m or 1.8 m above the ground). Eight villages were chosen as for the experiments. RESULTS: The use of one ASB station per house did not mark enough mosquitoes. Use of two and three stations per house gave feeding rates above the 25% goal. There was no statistical difference in the percentage of marked mosquitoes between two and three stations, however, the catches using two and three bait stations were both significantly higher than using one. There was no difference in An. gambiae s.l. feeding when stations were hung at 1.0 and 1.8 m. At 1.8 m stations sustained less accidental damage. ASB stations 1.8 m above ground were fed on by three of seven monitored insect orders. The monitored orders were: Hymenoptera, Lepidoptera, Coleoptera, Diptera, Hemiptera, Neuroptera and Orthoptera. Using one or two stations significantly reduced percentage of bait-fed NTOs compared to three stations which had the highest feeding rates. Percentages were as follows: 6.84 ± 2.03% Brachycera followed by wasps (Hymenoptera: Vespidae) 5.32 ± 2.27%, and Rhopalocera 2.22 ± 1.79%. Hanging the optimal number of stations per house for catching mosquitoes (two) at 1.8 m above ground, limited the groups of non-targets to Brachycera, Chironomidae, Noctuoidea, Rhopalocera, parasitic wasps and wasps (Hymenoptera). Feeding at 1.8 m only occurred when stations were damaged. CONCLUSIONS: The goal of marking quarter of the total Anopheles population per day was obtained using 2 bait stations at 1.8 m height above the ground. This configuration also had minimal effects on non-target insects.

Large-scale field trial of attractive toxic sugar baits (ATSB) for the control of malaria vector mosquitoes in Mali, West Africa.

BACKGROUND: The aim of this field trial was to evaluate the efficacy of attractive toxic sugar baits (ATSB) in Mali, where sustained malaria transmission occurs despite the use of long-lasting insecticidal nets (LLINs). ATSB bait stations were deployed in seven of 14 similar study villages, where LLINs were already in widespread use. The combined use of ATSB and LLINs was tested to see if it would substantially reduce parasite transmission by Anopheles gambiae sensu lato beyond use of LLINs alone. METHODS: A 2-day field experiment was conducted to determine the number of mosquitoes feeding on natural sugar versus those feeding on bait stations containing attractive sugar bait without toxin (ASB)-but with food dye. This was done each month in seven random villages from April to December 2016. In the following year, in seven treatment villages from May to December 2017, two ATSB bait stations containing the insecticide dinotefuran were placed on the outer walls of each building. Vector population density was evaluated monthly by CDC UV light traps, malaise traps, pyrethrum spray (PSCs) and human landing catches (HLCs). Female samples of the catch were tested for age by examination of the ovarioles in dissected ovaries and identification of Plasmodium falciparum sporozoite infection by ELISA. Entomological inoculation rates (EIR) were calculated, and reductions between treated and untreated villages were determined. RESULTS: In the 2-day experiment with ASB each month, there was a lower number of male and female mosquitoes feeding on the natural sugar sources than on the ASB. ATSB deployment reduced CDC-UV trap female catches in September, when catches were highest, were by 57.4% compared to catches in control sites. Similarly, malaise trap catches showed a 44.3% reduction of females in August and PSC catches of females were reduced by 48.7% in September. Reductions of females in HLCs were lower by 19.8% indoors and 26.3% outdoors in September. The high reduction seen in the rainy season was similar for males and reductions in population density for both males and females were > 70% during the dry season. Reductions of females with ≥ 3 gonotrophic cycles were recorded every month amounting to 97.1% in October and 100.0% in December. Reductions in monthly EIRs ranged from 77.76 to 100.00% indoors and 84.95% to 100.00% outdoors. The number of sporozoite infected females from traps was reduced by 97.83% at treated villages compared to controls. CONCLUSIONS: Attractive toxic sugar baits used against Anopheles mosquitoes in Mali drastically reduced the density of mosquitoes, the number of older females, the number of sporozoite infected females and the EIR demonstrating how ATSB significantly reduces malaria parasite transmission.

Is outdoor vector control needed for malaria elimination? An individual-based modelling study.

BACKGROUND: Residual malaria transmission has been reported in many areas even with adequate indoor vector control coverage, such as long-lasting insecticidal nets (LLINs). The increased insecticide resistance in Anopheles mosquitoes has resulted in reduced efficacy of the widely used indoor tools and has been linked with an increase in outdoor malaria transmission. There are considerations of incorporating outdoor interventions into integrated vector management (IVM) to achieve malaria elimination; however, more information on the combination of tools for effective control is needed to determine their utilization. METHODS: A spatial individual-based model was modified to simulate the environment and malaria transmission activities in a hypothetical, isolated African village setting. LLINs and outdoor attractive toxic sugar bait (ATSB) stations were used as examples of indoor and outdoor interventions, respectively. Different interventions and lengths of efficacy periods were tested. Simulations continued for 420 days, and each simulation scenario was repeated 50 times. Mosquito populations, entomologic inoculation rates (EIRs), probabilities of local mosquito extinction, and proportion of time when the annual EIR was reduced below one were compared between different intervention types and efficacy periods. RESULTS: In the village setting with clustered houses, the combinational intervention of 50% LLINs plus outdoor ATSBs significantly reduced mosquito population and EIR in short term, increased the probability of local mosquito extinction, and increased the time when annual EIR is less than one per person compared to 50% LLINs alone; outdoor ATSBs alone significantly reduced mosquito population in short term, increased the probability of mosquito extinction, and increased the time when annual EIR is less than one compared to 50% LLINs alone, but there was no significant difference in EIR in short term between 50% LLINs and outdoor ATSBs. In the village setting with dispersed houses, the combinational intervention of 50% LLINs plus outdoor ATSBs significantly reduced mosquito population in short term, increased the probability of mosquito extinction, and increased the time when annual EIR is less than one per person compared to 50% LLINs alone; outdoor ATSBs alone significantly reduced mosquito population in short term, but there were no significant difference in the probability of mosquito extinction and the time when annual EIR is less than one between 50% LLIN and outdoor ATSBs; and there was no significant difference in EIR between all three interventions. A minimum of 2 months of efficacy period is needed to bring out the best possible effect of the vector control tools, and to achieve long-term mosquito reduction, a minimum of 3 months of efficacy period is needed. CONCLUSIONS: The results highlight the value of incorporating outdoor vector control into IVM as a supplement to traditional indoor practices for malaria elimination in Africa, especially in village settings of clustered houses where LLINs alone is far from sufficient.

The invasive shrub Prosopis juliflora enhances the malaria parasite transmission capacity of Anopheles mosquitoes: a habitat manipulation experiment.

BACKGROUND: A neglected aspect of alien invasive plant species is their influence on mosquito vector ecology and malaria transmission. Invasive plants that are highly attractive to Anopheles mosquitoes provide them with sugar that is critical to their survival. The effect on Anopheles mosquito populations was examined through a habitat manipulation experiment that removed the flowering branches of highly attractive Prosopis juliflora from selected villages in Mali, West Africa. METHODS: Nine villages in the Bandiagara district of Mali were selected, six with flowering Prosopis juliflora, and three without. CDC-UV light traps were used to monitor their Anopheles spp. vector populations, and recorded their species composition, population size, age structure, and sugar feeding status. After 8 days, all of the flowering branches were removed from three villages and trap catches were analysed again. RESULTS: Villages where flowering branches of the invasive shrub Prosopis juliflora were removed experienced a threefold drop in the older more dangerous Anopheles females. Population density dropped by 69.4% and the species composition shifted from being a mix of three species of the Anopheles gambiae complex to one dominated by Anopheles coluzzii. The proportion of sugar fed females dropped from 73 to 15% and males from 77 to 10%. CONCLUSIONS: This study demonstrates how an invasive plant shrub promotes the malaria parasite transmission capacity of African malaria vector mosquitoes. Proper management of invasive plants could potentially reduce mosquito populations and malaria transmission.

Modelling optimum use of attractive toxic sugar bait stations for effective malaria vector control in Africa.

BACKGROUND: The development of insecticide resistance and the increased outdoor-biting behaviour of malaria vectors reduce the efficiency of indoor vector control methods. Attractive toxic sugar baits (ATSBs), a method targeting the sugar-feeding behaviours of vectors both indoors and outdoors, is a promising supplement to indoor tools. The number and configuration of these ATSB stations needed for malaria control in a community needs to be determined. METHODS: A hypothetical village, typical of those in sub-Saharan Africa, 600 × 600 m, consisting of houses, humans and essential resource requirements of Anopheles gambiae (sugar sources, outdoor resting sites, larval habitats) was simulated in a spatial individual-based model. Resource-rich and resource-poor environments were simulated separately. Eight types of configurations and different densities of ATSB stations were tested. Anopheles gambiae population size, human biting rate (HBR) and entomological inoculation rates (EIR) were compared between different ATSB configurations and densities. Each simulated scenario was run 50 times. RESULTS: Compared to the outcomes not altered by ATSB treatment in the control scenario, in resource-rich and resource-poor environments, respectively, the optimum ATSB treatment reduced female abundance by 98.22 and 91.80 %, reduced HBR by 99.52 and 98.15 %, and reduced EIR by 99.99 and 100 %. In resource-rich environments, n × n grid design, stations at sugar sources, resting sites, larval habitats, and random locations worked better in reducing vector population and HBRs than other configurations (P < 0.0001). However, there was no significant difference of EIR reductions between all ATSB configurations (P > 0.05). In resource-poor environments, there was no significant difference of female abundances, HBRs and EIRs between all ATSB configurations (P > 0.05). The optimum number of ATSB stations was about 25 for resource-rich environments and nine for resource-poor environments. CONCLUSIONS: ATSB treatment reduced An. gambiae population substantially and reduced EIR to near zero regardless of environmental resource availability. In resource-rich environments, dispersive configurations worked better in reducing vector population, and stations at or around houses worked better in preventing biting and parasite transmission. In resource-poor environments, all configurations worked similarly. Optimum numbers of bait stations should be adjusted according to seasonality when resource availability changes.

A spatial individual-based model predicting a great impact of copious sugar sources and resting sites on survival of Anopheles gambiae and malaria parasite transmission.

BACKGROUND: Agent-based modelling (ABM) has been used to simulate mosquito life cycles and to evaluate vector control applications. However, most models lack sugar-feeding and resting behaviours or are based on mathematical equations lacking individual level randomness and spatial components of mosquito life. Here, a spatial individual-based model (IBM) incorporating sugar-feeding and resting behaviours of the malaria vector Anopheles gambiae was developed to estimate the impact of environmental sugar sources and resting sites on survival and biting behaviour. METHODS: A spatial IBM containing An. gambiae mosquitoes and humans, as well as the village environment of houses, sugar sources, resting sites and larval habitat sites was developed. Anopheles gambiae behaviour rules were attributed at each step of the IBM: resting, host seeking, sugar feeding and breeding. Each step represented one second of time, and each simulation was set to run for 60 days and repeated 50 times. Scenarios of different densities and spatial distributions of sugar sources and outdoor resting sites were simulated and compared. RESULTS: When the number of natural sugar sources was increased from 0 to 100 while the number of resting sites was held constant, mean daily survival rate increased from 2.5% to 85.1% for males and from 2.5% to 94.5% for females, mean human biting rate increased from 0 to 0.94 bites per human per day, and mean daily abundance increased from 1 to 477 for males and from 1 to 1,428 for females. When the number of outdoor resting sites was increased from 0 to 50 while the number of sugar sources was held constant, mean daily survival rate increased from 77.3% to 84.3% for males and from 86.7% to 93.9% for females, mean human biting rate increased from 0 to 0.52 bites per human per day, and mean daily abundance increased from 62 to 349 for males and from 257 to 1120 for females. All increases were significant (P < 0.01). Survival was greater when sugar sources were randomly distributed in the whole village compared to clustering around outdoor resting sites or houses. CONCLUSIONS: Increases in densities of sugar sources or outdoor resting sites significantly increase the survival and human biting rates of An. gambiae mosquitoes. Survival of An. gambiae is more supported by random distribution of sugar sources than clustering of sugar sources around resting sites or houses. Density and spatial distribution of natural sugar sources and outdoor resting sites modulate vector populations and human biting rates, and thus malaria parasite transmission.

Indoor use of attractive toxic sugar bait (ATSB) to effectively control malaria vectors in Mali, West Africa.

BACKGROUND: Attractive toxic sugar bait (ATSB) solutions containing any gut toxins can be either sprayed on plants or used in simple bait stations to attract and kill sugar-feeding female and male mosquitoes. This field study in Mali demonstrates the effect of ATSB bait stations inside houses as a vector control method that targets and kills endophilic African malaria vectors. METHODS: The studies were conducted in five villages located near the River Niger, Mali. Baseline village-wide assessments of densities for female and male Anopheles gambiae sensu lato were performed by pyrethrum spray collections (PSC) in ten houses in each of five villages. To determine the rate of mosquito feeding on bait stations, one bait station per house containing attractive sugar bait (ASB) (without toxin) plus a food dye marker, was set up in ten houses in each of the five villages. PSC collections were conducted on the following day and the percentage of female and male mosquitoes that had fed was determined by visual inspection for the dye marker. Then, a 50-day field trial was done. In an experimental village, one bait station containing ATSB (1% boric acid active ingredient) was placed per bedroom (58 bedrooms), and indoor densities of female and male An. gambiae s.l. were subsequently determined by PSC, and female mosquitoes were age graded. RESULTS: In the five villages, the percentages of An. gambiae s.l. feeding inside houses on the non-toxic bait stations ranged from 28.3 to 53.1% for females and 36.9 to 78.3% for males. Following ATSB indoor bait station presentation, there was a significant reduction, 90% in female and 93% in male populations, of An. gambiae s.l. at the experimental village. A 3.8-fold decrease in the proportion of females that had undergone four or more gonotrophic cycles was recorded at the experimental village, compared to a 1.2-fold increase at the control village. CONCLUSION: The field trial demonstrates that An. gambiae s.l. feed readily from ATSB bait stations situated indoors, leading to a substantial reduction in the proportion of older female mosquitoes. This study demonstrates that ATSB inside houses can achieve impressive malaria vector control in Africa.

Control of Aedes albopictus with attractive toxic sugar baits (ATSB) and potential impact on non-target organisms in St. Augustine, Florida.

The purpose of this study was to test the efficacy of bait stations and foliar applications containing attractive toxic sugar baits (ATSB) and eugenol to control Aedes albopictus. At the same time, the potential impact of these control methods was evaluated on non-target organisms. The study was conducted at five tire sites in St. Augustine, Florida. A. albopictus populations were significantly reduced with ATSB-eugenol applications applied directly to non-flowering vegetation and as bait stations compared with non-attractive sugar baits and control. The application of ATSB made to non-flowering vegetation resulted in more significant reductions of mosquito populations compared to the application of ATSB presented in a bait station. Over 5.5% of the non-targets were stained in the flowering vegetation application site. However, when the attractive sugar bait application was made to non-flowering vegetation or presented in bait stations, the impact on non-target insects was very low for all non-target orders as only 0.6% of the individual insects were stained with the dye from the sugar solutions, respectively. There were no significant differences between the staining of mosquitoes collected in flowering vegetation (206/1000) or non-flowering vegetation (242/1000) sites during the non-target evaluation. Our field studies support the use of eugenol as an active ingredient for controlling the dengue vector A. albopictus when used as an ATSB toxin and demonstrates potential use in sub-tropical and tropical environments for dengue control.

Quantifying the mosquito's sweet tooth: modelling the effectiveness of attractive toxic sugar baits (ATSB) for malaria vector control.

BACKGROUND: Current vector control strategies focus largely on indoor measures, such as long-lasting insecticide treated nets (LLINs) and indoor residual spraying (IRS); however mosquitoes frequently feed on sugar sources outdoors, inviting the possibility of novel control strategies. Attractive toxic sugar baits (ATSB), either sprayed on vegetation or provided in outdoor bait stations, have been shown to significantly reduce mosquito densities in these settings. METHODS: Simple models of mosquito sugar-feeding behaviour were fitted to data from an ATSB field trial in Mali and used to estimate sugar-feeding rates and the potential of ATSB to control mosquito populations. The model and fitted parameters were then incorporated into a larger integrated vector management (IVM) model to assess the potential contribution of ATSB to future IVM programmes. RESULTS: In the Mali experimental setting, the model suggests that about half of female mosquitoes fed on ATSB solution per day, dying within several hours of ingesting the toxin. Using a model incorporating the number of gonotrophic cycles completed by female mosquitoes, a higher sugar-feeding rate was estimated for younger mosquitoes than for older mosquitoes. Extending this model to incorporate other vector control interventions suggests that an IVM programme based on both ATSB and LLINs may substantially reduce mosquito density and survival rates in this setting, thereby substantially reducing parasite transmission. This is predicted to exceed the impact of LLINs in combination with IRS provided ATSB feeding rates are 50% or more of Mali experimental levels. In addition, ATSB is predicted to be particularly effective against Anopheles arabiensis, which is relatively exophilic and therefore less affected by IRS and LLINs. CONCLUSIONS: These results suggest that high coverage with a combination of LLINs and ATSB could result in substantial reductions in malaria transmission in this setting. Further field studies of ATSB in other settings are needed to assess the potential of ATSB as a component in future IVM malaria control strategies.

Attractive toxic sugar bait (ATSB) methods decimate populations of Anopheles malaria vectors in arid environments regardless of the local availability of favoured sugar-source blossoms.

BACKGROUND: Attractive toxic sugar bait (ATSB) methods are a new and promising "attract and kill" strategy for mosquito control. Sugar-feeding female and male mosquitoes attracted to ATSB solutions, either sprayed on plants or in bait stations, ingest an incorporated low-risk toxin such as boric acid and are killed. This field study in the arid malaria-free oasis environment of Israel compares how the availability of a primary natural sugar source for Anopheles sergentii mosquitoes: flowering Acacia raddiana trees, affects the efficacy of ATSB methods for mosquito control. METHODS: A 47-day field trial was conducted to compare impacts of a single application of ATSB treatment on mosquito densities and age structure in isolated uninhabited sugar-rich and sugar-poor oases relative to an untreated sugar-rich oasis that served as a control. RESULTS: ATSB spraying on patches of non-flowering vegetation around freshwater springs reduced densities of female An. sergentii by 95.2% in the sugar-rich oasis and 98.6% in the sugar-poor oasis; males in both oases were practically eliminated. It reduced daily survival rates of female An. sergentii from 0.77 to 0.35 in the sugar-poor oasis and from 0.85 to 0.51 in the sugar-rich oasis. ATSB treatment reduced the proportion of older more epidemiologically dangerous mosquitoes (three or more gonotrophic cycles) by 100% and 96.7%, respectively, in the sugar-poor and sugar-rich oases. Overall, malaria vectorial capacity was reduced from 11.2 to 0.0 in the sugar-poor oasis and from 79.0 to 0.03 in the sugar-rich oasis. Reduction in vector capacity to negligible levels days after ATSB application in the sugar-poor oasis, but not until after 2 weeks in the sugar-rich oasis, show that natural sugar sources compete with the applied ATSB solutions. CONCLUSION: While readily available natural sugar sources delay ATSB impact, they do not affect overall outcomes because the high frequency of sugar feeding by mosquitoes has an accumulating effect on the probability they will be attracted to and killed by ATSB methods. Operationally, ATSB methods for malaria vector control are highly effective in arid environments regardless of competitive, highly attractive natural sugar sources in their outdoor environment.

Field experiments of Anopheles gambiae attraction to local fruits/seedpods and flowering plants in Mali to optimize strategies for malaria vector control in Africa using attractive toxic sugar bait methods.

BACKGROUND: Based on recent studies in Israel demonstrating that attractive toxic sugar bait (ATSB) methods can be used to decimate local anopheline and culicine mosquito populations, an important consideration is whether the same methods can be adapted and improved to attract and kill malaria vectors in Africa. The ATSB approach uses fruit or flower scent as an attractant, sugar solution as a feeding stimulant, and an oral toxin. The ATSB solutions are either sprayed on vegetation or suspended in simple bait stations, and the mosquitoes ingesting the toxic solutions are killed. As such, this approach targets sugar-feeding female and male mosquitoes. This study examines the attractiveness of African malaria vectors to local fruits/seedpods and flowering plants, key biological elements of the ATSB approach for mosquito control. METHODS: Three field experiments were conducted at sites in Mali. The attraction of Anopheles gambiae s.l. to 26 different local fruits and seedpods was determined at a site in the semi-arid Bandiagara District of Mali. Wire mesh glue traps with fruits/seedpods suspended on skewers inside were set along a seasonal lagoon. Seven replicates of each fruit/seedpod species were tested, with a water-soaked sponge and a sugar-soaked sponge as controls. The attraction of An. gambiae s.l. to 26 different types of flowering plants was determined at a site near Mopti in Mali. The flowering plants held in a water-filled buried container were tested using the same glue traps, with controls including water only and sugar solution. Six replicates of each selected plant type were tested on transects between rice paddies. Additional studies using CDC light traps were done to determine the relative densities and periodicity of An. gambiae s.l. attraction to branches of the most highly attractive flowering plant, branches without flowers, human odor, and candescent light. RESULTS: Of the 26 fruits and seedpods tested, 6 were attractive to An. gambiae s.l. females and males, respectively. Guava (Psidium guajava) and honey melon (Cucumis melo) were the two most attractive fruits for both females and males. Of the 26 flowering plants tested, 9 were significantly attractive for females, and 8 were attractive for males. Acacia macrostachya was the most attractive flowering plant. Periodicity studies using this plant showed peaks of An. gambiae s.l. attraction between 1930 and 2200 h and 0400-0500 h, which differed considerably from the response to human odors, which expectedly peaked at around midnight. CONCLUSION: These field experiments in Mali highlight that female and male An. gambiae s.l. have pronounced differences in attraction for diverse types of indigenous fruits/seedpods and flowering plants. The identification of attractive fruits and seedpods shows that a variety of indigenous and locally abundant natural products could potentially be used as juices to make ATSB solution for mosquito control. As well, the simple methods used to identify the most attractive flowering plants provide valuable insights into the natural history of sugar feeding for An. gambiae s.l. These observations can be used to guide future strategies for employing ATSB methods for malaria vector control in Africa. They also provide a basis for subsequent chemical analysis and development of attractive baits for mosquito control.

Successful field trial of attractive toxic sugar bait (ATSB) plant-spraying methods against malaria vectors in the Anopheles gambiae complex in Mali, West Africa.

BACKGROUND: Based on highly successful demonstrations in Israel that attractive toxic sugar bait (ATSB) methods can decimate local populations of mosquitoes, this study determined the effectiveness of ATSB methods for malaria vector control in the semi-arid Bandiagara District of Mali, West Africa. METHODS: Control and treatment sites, selected along a road that connects villages, contained man-made ponds that were the primary larval habitats of Anopheles gambiae and Anopheles arabiensis. Guava and honey melons, two local fruits shown to be attractive to An. gambiae s.l., were used to prepare solutions of Attractive Sugar Bait (ASB) and ATSB that additionally contained boric acid as an oral insecticide. Both included a color dye marker to facilitate determination of mosquitoes feeding on the solutions. The trial was conducted over a 38-day period, using CDC light traps to monitor mosquito populations. On day 8, ASB solution in the control site and ATSB solution in the treatment site were sprayed using a hand-pump on patches of vegetation. Samples of female mosquitoes were age-graded to determine the impact of ATSB treatment on vector longevity. RESULTS: Immediately after spraying ATSB in the treatment site, the relative abundance of female and male An. gambiae s.l. declined about 90% from pre-treatment levels and remained low. In the treatment site, most females remaining after ATSB treatment had not completed a single gonotrophic cycle, and only 6% had completed three or more gonotrophic cycles compared with 37% pre-treatment. In the control site sprayed with ASB (without toxin), the proportion of females completing three or more gonotrophic cycles increased from 28.5% pre-treatment to 47.5% post-treatment. In the control site, detection of dye marker in over half of the females and males provided direct evidence that the mosquitoes were feeding on the sprayed solutions. CONCLUSION: This study in Mali shows that even a single application of ATSB can substantially decrease malaria vector population densities and longevity. It is likely that ATSB methods can be used as a new powerful tool for the control of malaria vectors, particularly since this approach is highly effective for mosquito control, technologically simple, inexpensive, and environmentally safe.

Frequent sugar feeding behavior by Aedes aegypti in Bamako, Mali makes them ideal candidates for control with attractive toxic sugar baits (ATSB).

BACKGROUND: Current tools and strategies are not sufficient to reliably address threats and outbreaks of arboviruses including Zika, dengue, chikungunya, and yellow fever. Hence there is a growing public health challenge to identify the best new control tools to use against the vector Aedes aegypti. In this study, we investigated Ae. aegypti sugar feeding strategies in Bamako, Mali, to determine if this species can be controlled effectively using attractive toxic sugar baits (ATSB). METHODOLOGY: We determined the relative attraction of Ae. aegypti males and females to a variety of sugar sources including flowers, fruits, seedpods, and honeydew in the laboratory and using plant-baited traps in the field. Next, we observed the rhythm of blood feeding versus sugar feeding activity of Ae. aegypti in vegetation and in open areas. Finally, we studied the effectiveness of spraying vegetation with ATSB on Ae. aegypti in sugar rich (lush vegetation) and in sugar poor (sparse vegetation) urban environments. PRINCIPAL FINDINGS: Male and female laboratory sugar feeding rates within 24 h, on 8 of 16 plants offered were over 80%. The survival rates of mosquitoes on several plant sources were nearly as long as that of controls maintained on sucrose solution. In the field, females were highly attracted to 11 of 20 sugar sources, and 8 of these were attractive to males. Peak periods of host attraction for blood-feeding and sugar feeding in open areas were nearly identical and occurred shortly after sunrise and around sunset. In shaded areas, the first sugar-seeking peak occurred between 11:30 and 12:30 while the second was from 16:30 to 17:30. In a 50-day field trial, ATSB significantly reduced mean numbers of landing / biting female Ae. aegypti in the two types of vegetation. At sugar poor sites, the mean pre-treatment catch of 20.51 females on day 14 was reduced 70-fold to 0.29 on day 50. At sugar rich sites, the mean pre-treatment catch of 32.46 females on day 14 was reduced 10-fold to a mean of 3.20 females on day 50. CONCLUSIONS: This is the first study to show how the vector Ae. aegypti depends on environmental resources of sugar for feeding and survival. The demonstration that Ae. aegypti populations rapidly collapsed after ATSB treatment, in both sugar rich and sugar poor environments, is strong evidence that Ae. aegypti is sugar-feeding frequently. Indeed, this study clearly demonstrates that Ae. aegypti mosquitoes depend on natural sugar resources, and a promising new method for vector control, ATSB, can be highly effective in the fight against Aedes-transmitted diseases.

Efficacy of toxic sugar baits against adult cistern-dwelling Anopheles claviger.

Sugar baits were used for mosquito control in cisterns at the building complex of Mar Saba Monastery in the Judean Hills, Israel. These cisterns provide fresh water for the monastery and are breeding and resting sites for Anopheles claviger (Meigen). The baits, including sucrose, fruit juice, dye marker and oral insecticide, were presented at the entrance of the cisterns and caused a drastic decline of A. claviger numbers in the neighbourhood. Pre-treatment catches of 16.8+/-4.0 to 33+/-8.1 females per trap decreased to between 2.5+/-1.3 and 0.3+/-0.6 females per trap six nights after treatment. Numbers of males were initially 9.2+/-1.7 to 21.0+/-4.8 specimens per trap and decreased to between 1.0+/-0.4 and 0.5+/-0.2 per trap post treatment. In the control area, where baits without toxin were presented in cisterns, there were only small fluctuations in the mosquito population. Non-poisonous bait marked >95% of the mosquitoes exiting from a cistern in the control area. Following treatment, the number of human-landing mosquitoes decreased by more than ten-fold in the experimental area, whilst in the control area there was no significant decrease.

An approach to mosquito control: using the dominant attraction of flowering Tamarix jordanis trees against Culex pipiens.

In this study, we identified blossoms that attract Culex pipiens L. s.l. in a Mediterranean habitat by using branches of 26 common plant species as baits for traps. The highest catch, 60.5% of the total, by flowers of Tamarix jordanis Boiss., was approximately 6 times greater than the 10.7% caught by flowering Polygonum equisetiforme Sm., and 10 times higher than the 6.6% caught by flowers of Acacia saligna (Lindle) H. L. Wendl. The catch elicited by the other plants ranged between 4.0 and 0.1%. Plant attraction also was evaluated in a field situation. Experimental and control sites were similar strips of vegetation along water channels with T. jordanis trees in the center. In the experimental site, these trees were sprayed with sucrose solution, food dye, and oral insecticide (Spinosad). Concurrently, patches of plant species and trees in the control site were sprayed with solutions of sucrose and different food dye markers. Cx. pipiens populations in both sites were monitored. The highest proportion (65.2%) of the marked mosquitoes in the control site carried the dye of flowering T. jordanis. The dye of flowering P. equisetiforme and that of A. saligna were found, respectively, in 8.1 and 3.5% of the labeled mosquitoes. The marker of reed groups (Phragmites australis [Cav.] Steudel) above the water was found in 19.4% of mosquitoes, whereas the different marker of dry land reeds was found in only 0.4% of the labeled mosquitoes. In the experimental site, after treatment, the mosquitoes decreased from approximately 255 per trap to approximately 24 mosquitoes per trap, whereas the catch in the control site reached approximately 400 mosquitoes per trap.

Decline of Anopheles sergentii and Aedes caspius populations following presentation of attractive toxic (spinosad) sugar bait stations in an oasis.

The effect of attractive sugar bait stations, including sucrose, juice of nectarine, slow-release substances, preservatives, red food-dye marker, and the oral insecticide spinosad, on Anopheles sergentii and Aedes caspius populations was studied in a small oasis in a southern desert of Israel. Feeding on similar baits without an insecticide was monitored as a control in a similar neighboring oasis. The insecticide caused a drastic decrease in the number of mosquitoes. Compared to the control site, the An. sergentii population was reduced to less than a tenth and that of Ae. caspius declined to a third. The majority of the mosquitoes, 76.0% of An. sergentii females and 74.8% of Ae. caspius females, were marked by the food dye in the control site.

Indoor protection against mosquito and sand fly bites: a comparison between citronella, linalool, and geraniol candles.

The repellent effect of 3 essential-oil-based candles was evaluated in a high biting pressure environment in Israel. In human landing assays, the repellency rate of 5% citronella candles against mosquitoes was 29.0%, of 5% linalool candles was 71.1%, and of 5% geraniol candles was 85.4%. The candles with geraniol were about twice as effective as those with linalool and were about 5 times as effective as citronella candles in protecting a person from being bitten indoors by mosquitoes. The repellency rate of 5% citronella candles towards sand flies was 24.7%, of 5% linalool candles was 55.2%, and of 5% geraniol candles was 79.7%. A geraniol candle was almost 5 times as effective as a citronella candle and about twice as effective as a linalool candle in protecting a person from being bitten indoors by sand flies.

Ability of essential oil candles to repel biting insects in high and low biting pressure environments.

The first goal of this study was to compare the degree of personal protection against biting insects provided by geraniol, linalool, and citronella candle (5%) vapors outdoors, where such products are commonly used. At a distance of 1.0 m, citronella candles reduced the number of female mosquitoes caught in Centers for Disease Control and Prevention traps by 35.4% and sand flies by 15.4%, linalool candles reduced female mosquitoes by 64.9% and sand flies by 48.5%, while geraniol candles reduced female mosquitoes by 81.5% and sand flies by 69.8%. By increasing the distance to 2 m and 3 m, the repellency dropped significantly. The second goal was to compare the degree of personal protection provided by the best performing candle, geraniol, under conditions of high and low biting pressure. The introduction of geraniol candles to protect volunteers in a high biting pressure environment reduced the mosquito pressure by an average of 56% and the sand fly pressure by 62% over a distance of 1.0 m. In the low biting pressure environment, geraniol reduced the mosquito pressure by an average of 62%. No sand flies were present at this site.

Glycolytic and chitinolytic activities of Phlebotomus papatasi (Diptera: Psychodidae) from diverse ecological habitats.

The sand fly Phlebotomus papatasi Scopoli, 1786, the vector of Leishmania major Yakimoff et Schokhor, 1914, is found in desert areas where sugars are scarce but also in habitats that abound in sugar sources. The sand flies require sugar meals from plant sources for their energy requirements and to hydrolyze these complex sugars, they need a repertoire of glycosidases. We presumed that there are differences in the levels of glycosidase activities in flies from such habitats and also assumed that they may be instrumental in modulating the flies' susceptibility to L. major infections. Phlebotomus papatasi originating from diverse ecological habitats ranging from an oasis to desert sites were colonized. They were analyzed for weight changes and glycosidase activities before and after feeding on 1M sucrose solution. Oasis flies were smaller than desert flies but took larger sugar meals. Homogenates of these flies hydrolyzed 16 synthetic and 2 natural glycoside substrates to varying degrees. The arid-region flies tended to produce more glycosidase activity than those originating in sugar-rich environments, especially sucrase, alpha- and beta-glucosidase, aalpha-fucosidase, alpha-mannosidase, and alpha- and beta-N-acetylgalactosaminidase. However, chitinolytic enzyme activities and particularly the beta-N-acetylhexosaminidase activity of oasis flies were higher than other flies tested. In comparing the desert flies, there were also significant differences in glycolytic enzyme activities between the spring-line (flowering season) of flies and the autumn-line (end of dry season) flies. A range of saccharide inhibitors was tested to demonstrate the specificity of the enzymes.

A novel window entry/exit trap for the study of endophilic behavior of mosquitoes.

To overcome the limitations of currently used window entry/exit traps, we developed an efficient new glue net entry/exit trap (GNT) that is economical, easily transported and assembled, and can be utilized for a variety of studies which aim to investigate the dynamics of mosquito movements between indoor and outdoor habitats. Cage experiments were conducted to determine what percentage of mosquitoes trying to pass through the netting are actually being caught. The GNT caught 97% of female and 98% of male Anopheles gambiae s.s., as well as 97.5% of female and 98% of male Culex pipiens attempting to cross into a bait chamber adjoining the release cage. During a six day field study, the bedroom windows of 12 homes in Mali were fitted with entry/exit GNTs. Traps without glue were fitted over the inside and outside bedroom windows of an additional 12 homes as a control. A total of 446 An. gambiae s.l. were caught attempting to exit dwellings while 773 An. gambiae s.l. were caught attempting to enter. The number of males and females attempting to exit dwellings were roughly similar (215 and 231 respectively) while there was a slight difference in the number of males and females trying to enter (382 and 430 respectively). Pyrethrum spray catches (PSC's) conducted inside the dwellings on the last day of the experiment yielded only six females and a single male.