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

The efficacy of attractive toxic sugar baits (ATSB) with the active ingredient eugenol, an Environmental Protection Agency exempt compound, was evaluated against vector and nuisance mosquitoes in both laboratory and field studies. In the laboratory, eugenol combined in attractive sugar bait (ASB) solution provided high levels of mortality for Aedes aegypti, Culex quinquefasciatus, and Anopheles quadrimaculatus. Field studies demonstrated significant control: > 70% reduction for Aedes atlanticus, Ae. infirmatus, and Culex nigripalpus and > 50% reduction for An. crucians, Uranotaenia sapphirina, Culiseta melanura, and Cx. erraticus three weeks post ATSB application. Furthermore, non-target feeding of six insect orders, Hymenoptera, Lepidoptera, Coleoptera, Diptera, Hemiptera, and Orthoptera, was evaluated in the field after application of a dyed-ASB to flowering and non-flowering vegetation. ASB feeding (staining) was determined by dissecting the guts and searching for food dye with a dissecting microscope. The potential impact of ATSB on non-targets, applied on green non-flowering vegetation was low for all non-target groups (0.9%). However, application of the ASB to flowering vegetation resulted in significant staining of the non-target insect orders. This highlights the need for application guidelines to reduce non-target effects. No mortality was observed in laboratory studies with predatory non-targets, spiders, praying mantis, or ground beetles, after feeding for three days on mosquitoes engorged on ATSB. Overall, our laboratory and field studies support the use of eugenol as an active ingredient for controlling important vector and nuisance mosquitoes when used as an ATSB toxin. This is the first study demonstrating effective control of anophelines in non-arid environments which suggest that even in highly competitive sugar rich environments this method could be used for control of malaria in Latin American countries.

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. Aedes 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 Ae. albopictus when used as an ATSB toxin and demonstrates potential use in sub-tropical and tropical environments for dengue control.

We evaluated the efficacy of attractive toxic sugar baits (ATSB) in the laboratory and field with the low-risk active ingredient dinotefuran against mosquito populations. Preliminary laboratory assays indicated that dinotefuran in solution with the sugar baits was ingested and resulted in high mortality of female Culex quinquefasciatus Say and Aedes aegypti Linnaeus. Field studies demonstrated >70% reduction of mosquito populations at 3 wk post-ATSB application. Nontarget feeding of seven insect orders—Hymenoptera, Lepidoptera, Coleoptera, Diptera, Hemiptera, Orthoptera, and Neuroptera—was evaluated in the field after application of attractive sugar baits (ASB) on vegetation by dissecting the guts and searching for food dye with a dissecting microscope. Nontargets were found stained with ASB 0.9% of the time when the application was applied on green nonflowering vegetation. Only two families were significantly impacted by the ASB application: Culicidae (mosquitoes) and Chironomidae (nonbiting midges) of the order Diptera. Pollinators of the other insect orders were not significantly impacted. No mortality was observed in the laboratory studies with predatory nontargets, wolf spiders or ground beetles, after feeding for 3 d on mosquitoes engorged on ATSB applied to vegetation. Overall, this novel control strategy had little impact on nontarget organisms, including pollinators and beneficial insects, and was effective at controlling mosquito populations, further supporting the development of ATSB for commercial use.

Attractive toxic sugar baits (ATSBs) were used to control mosquitoes in the storm drains of a residential area on the outskirts of St Augustine, Florida. The drainage system was newly constructed and no mosquitoes were breeding inside it. The area covered by the storm drains was divided in half; 10 drains served as control drains and 16 drains served as experimental drains. The baits, which consisted of a mixture of brown sugar, fruit juice, green dye marker and boric acid, were presented at the entrances of the treated drains and exit traps were positioned over the drain openings and the connecting tubes leading to retention ponds. Similar baits with orange dye and without toxin were presented at the entrances of control drains. A total of 220 pupae of Culex quinquefasciatus (Diptera: Culicidae) were released in each control and toxin-treated drain, and the numbers of recovered mosquitoes were examined to determine the effectiveness of ATSBs in the storm drain system. An average of 178.2 mosquitoes exited each drain in the control area; 87.0% of these had fed on the baits and were stained orange, whereas 13.0% were unstained. In the toxin-treated drains, 83.7% of hatched females and 86.6% of hatched males were controlled by the baits.