Point Protection with Transfluthrin against Musca domestica L. in a Semi-Field Enclosure.

House flies are notoriously difficult to control, owing to their tendency to live in close relationships with humans and their livestock, and their rapid development of resistance to chemical controls. With this in mind, we explored an alternative chemical control, a spatial repellent to deter Musca domestica L. from points we wanted to protect (i.e., a baited trap). Our results demonstrated that the synthetic spatial repellent, transfluthrin, is effective in preventing M. domestica adults from entering protected traps for both a susceptible strain (CAR21) and a field-acquired permethrin-resistant strain (WHF; 24 h LD50 resistance ratio of 150), comprising 22% and 28% of the total number of flies collected, respectively. These results are promising and demonstrate that transfluthrin can be an effective spatial repellent to protect points of interest where needed.

Comparison of Trap and Equine Attraction to Mosquitoes.

Mosquitoes are pests of horses, but mosquito trap efficacy data, especially the ability of traps to protect horses, are lacking. Studies were conducted to investigate the comparative attraction between traps and horses, increase trap attraction by adding horse odors to the airstream of a trap, determine the spatial distribution of adult mosquitoes, estimate the numbers of mosquitoes feeding on horses, determine the relative attraction of horses to mosquitoes, and estimate the range of mosquitoes' attraction between two horses. When a horse and a mosquito trap were placed 3.5 m apart, there was a significant reduction in mosquitoes entering the trap. Adding horse odors to the airstream of a trap produced equivocal results because the horse providing the odors influenced the trap catches. Mosquitoes were not evenly distributed across the study site, which emphasized the importance of trap placement. Vacuuming mosquitoes from the horses in different seasons demonstrated that 324 and 359 mosquitoes per hour were feeding during the two studies. Separate analysis of data from the two horses vacuumed simultaneously revealed that one horse attracted twice as many mosquitoes as the other. This caused the results of a study to determine the attraction range of two horses moved from 3.5 to 20.4 m apart to be inconclusive.

Semi-field evaluation of a novel controlled release device using transfluthrin as spatial repellent to prevent entry of mosquitoes into military tents.

Mosquitoes can impact military operational readiness by transmission of disease-causing pathogens or through secondary effects, e.g., annoyance and bites. The focus of this research was to determine if an array of novel controlled release passive devices (CRPD) utilizing the spatial repellent, transfluthrin (TF), as the active ingredient could prevent entry of mosquitoes into military tents for up to 4 weeks. The TF-charged CRPDs were spaced along six strands of monofilament and hung across the tent entrance. Efficacy was evaluated with caged Aedes aegypti to indicate knockdown/mortality effects, and four species of free-flying mosquitoes, Ae. aegypti, Aedes taeniorhynchus, Anopheles quadrimaculatus and Culex quinquefasciatus, to indicate repellent effects. Bioassay cages containing Ae. aegypti were hung vertically at 0.5, 1.0 and 1.5 â€‹m above ground level at designated locations inside of the tents. Knockdown/mortality counts were made every 15 min for the first hour, then at 2, 4 and 24 h post-exposure. Free fliers were recaptured in BG traps operated from 4 to 24 h post-exposure. Knockdown/mortality was gradual until 4 h post-exposure. This increased to near 100% by 24 h in the treated tent but was < 2% in the control tent. There was a significant reduction in the recapture rates of all free-flying species in the treated tent compared with the control tent. Results indicate that TF-charged CRPDs can significantly reduce the numbers of mosquitoes entering military tents and that the four species were affected similarly by the TF. The needs for additional research are discussed.

Studies on the Volatiles Composition of Stored Sheep Wool, and Attractancy toward Aedes aegypti Mosquitoes.

To discover new natural materials for insect management, commercially available stored sheep wool was investigated for attractancy to female adult Aedes aegypti mosquitoes. The volatiles from sheep wool were collected by various techniques of headspace (HS) extractions and hydrodistillation. These extracts were analyzed using gas chromatography-mass spectrometry (GC-MS) and gas chromatography-flame ionization detector (GC-FID) coupled with GC-MS. Fifty-two volatile compounds were detected, many of them known for their mosquito attractant activity. Seven compounds were not previously reported in sheep products. The volatile composition of the extracts varied significantly across collections, depending on the extraction techniques or types of fibers applied. Two types of bioassay were conducted to study attractancy of the sheep wool volatiles to mosquitoes: laboratory bioassays using glass tubes, and semi-field bioassays using large, screened outdoor cages. In bioassays with glass tubes, the sheep wool hydrodistillate and its main component, thialdine, did not show any significant attractant activity against female adult Ae. aegypti mosquitoes. Semi-field bioassays in two large screened outdoor cages, each equipped with a U.S. Centers for Disease Control (CDC) trap and the various bait setups with Vortex apparatus, revealed that vibrating wool improved mosquito catches compared to the setups without wool or with wool but not vibrating. Sheep wool, when vibrated, may release intensively volatile compounds, which could serve as olfactory cues, and play significant role in making the bait attractive to mosquitoes. Sheep wool is a readily available, affordable, and environment-friendly material. It should have the potential to be used as a mosquito management and surveillance component in dynamic bait setups.

Spatial Repellents Protect Small Perimeters from Riceland Mosquitoes in a Warm-Humid Environment.

Recent experiments suggest spatial repellents may significantly reduce biting pressure from host-seeking riceland mosquitoes, such as Anopheles quadrimaculatus, in a warm-humid open-field habitat. However, little is known regarding efficacy of these formulations in partially enclosed spaces where US military personnel may be sheltered or concealed in an operational environment. In this study we investigated the capability of 3 spatial repellents-metofluthrin, linalool, and d-cis/trans allethrin-to reduce mosquito incursion into small open-top enclosures of US military camouflage netting. We found that metofluthrin was more effective in partially enclosed spaces compared with the open field, whereas both linalool and d-cis/trans allethrin provided superior protection in the open. These findings support strategic selection of spatial repellents depending on the environment immediately surrounding the host.

Olfactometric Comparison of the Volatile Insecticide, Metofluthrin, Through Behavioral Responses of Aedes albopictus (Diptera: Culicidae).

Testing behavioral response to insecticidal volatiles requires modifications to the existing designs of olfactometers. To create a testing apparatus in which there is no chemical memory to confound tests, we detail the technical aspects of a new tool with design influences from other olfactometry tools. In addition, this new tool was used to evaluate a novel formulation of metofluthrin for use as an outdoor residual treatment. After sourcing materials to prioritize glass and metal construction, a modular wind tunnel was developed that hybridizes wind tunnel and olfactometer specifications. Volatile contaminants were removed by strong ultraviolet light within the chamber before and between trials. Repellent trials were conducted with an experimental formulation of metofluthrin and a commercial formulation of esfenvalerate, prallethrin, and piperonyl butoxide (Onslaught Fast Cap) against Aedes albopictus (Skuse). Toxicant vapors were delivered with attractants from a lure with behavioral responses scored 20 min post-exposure. Upwind attraction to the attractant lure and the Onslaught Fast Cap plus lure resulted in 90 and 75% capture, respectively. In contrast, metofluthrin vapors resulted in less than 10% attraction, while also causing repellency, disorientation, knockdown, and mortality effects. Our findings demonstrated that an inert modular wind tunnel was functional for mitigating toxic secondary exposures of spatial repellents amidst complex behavioral analysis in mosquitoes. The resulting observations with formulated metofluthrin positively reinforce the merit of transitioning metofluthrin into expanded roles in mosquito management.

A new approach to improve acoustic trapping effectiveness for Aedes aegypti (Diptera: Culicidae).

Monitoring mosquito populations is essential to designing and implementing control strategies. Recent strategies based on releasing biologically modified mosquitoes have increased the need to effectively monitor mosquito abundance. Unfortunately, existing surveillance traps are of limited value due to their high cost and low capture rates. Here, we report the results of experiments designed to evaluate the effectiveness of an acoustic trap prototype. Stimuli synthesized from recordings of Ae. aegypti wingbeat signals and pure tones were evaluated as attractants to males in indoor and semi-field conditions. Overall, the acoustic trap´s efficacy differed significantly between indoor and semi-field conditions. After two hours of indoor recapture, ∼69% of males were collected from acoustic traps broadcasting pure tones while ∼78% of males were collected using synthesized wingbeat signals. Under semi-field conditions, however, acoustic traps collected less than ∼1.7% of the males released. Increasing the intensity of the signals up to 90 dB (SPL re. 20 uPa at 1 m from the trap) did not improve the capture rate under semi-field conditions. Overall, our results indicate that acoustic signals synthesized from recordings of wingbeats can be used to enhance capture of male Ae. aegypti.

A Low-Cost, Passive Release Device for the Surveillance and Control of Mosquitoes.

Mosquitoes continue to be a major threat to global health, and the ability to reliably monitor, catch, and kill mosquitoes via passive traps is of great importance. Global, low-cost, and easy-to-use outdoor devices are needed to augment existing efforts in mosquito control that combat the spread of disease, such as Zika. Thus, we have developed a modular, portable, non-powered (passive), self-contained, and field-deployable device suitable for releasing volatiles with a wide range of applications such as attracting, repelling, and killing mosquitoes. This unique device relies on a novel nested wick and two-reservoir design that achieves a constant release of volatiles over several hundred hours. Devices loaded with one of either two compounds, geraniol or 1-methylpiperazine (MP), were tested in a controlled environment (32 °C and 70% relative humidity), and both compounds achieved a constant release from our devices at a rate of 2.4 mg/h and 47 mg/h, respectively. The liquid payload can be volatile attractants or repellants as well as mosquitocide-containing feeding solutions for capture and surveillance. This low-cost device can be utilized for both civilian and military mosquito control purposes, but it will be particularly important for protecting those in economically repressed environments, such as sub-Saharan Africa and Central and South America.

A combined experimental-computational approach for spatial protection efficacy assessment of controlled release devices against mosquitoes (Anopheles).

This work describes the use of entomological studies combined with in silico models (computer simulations derived from numerical models) to assess the efficacy of a novel device for controlled release of spatial repellents. Controlled Release Devices (CRDs) were tested with different concentrations of metofluthrin and tested against An. quadrimaculatus mosquitoes using arm-in cage, semi-field, and outdoor studies. Arm-in-cage trials showed an approximate mean values for mosquito knockdown of 40% and mosquito bite reduction of 80% for the optimal metofluthrin formulation for a 15-minute trial. Semi-field outdoor studies showed a mean mortality of a 50% for 24 hour trial and 75% for a 48 hour trial for optimal concentrations. Outdoors studies showed an approximate mean mortality rate of 50% for a 24 hour trial for optimal concentrations. Numerical simulations based on Computational Fluid Dynamics (CFD) were performed in order to obtain spatial concentration profiles for 24 hour and 48 hour periods. Experimental results were correlated with simulation results in order to obtain a functional model that linked mosquito mortality with the estimated spatial concentration for a given period of time. Such correlation provides a powerful insight in predicting the effectiveness of the CRDs as a vector-control tool. While CRDs represent an alternative to current spatial repellent delivery methods, such as coils, candles, electric repellents, and passive emanators based on impregnated strips, the presented method can be applied to any spatial vector control treatment by correlating entomological endpoints, i.e. mortality, with in-silico simulations to predict overall efficacy. The presented work therefore presents a new methodology for improving design, development and deployment of vector-control tools to reduce transmission of vector-borne diseases, including malaria and dengue.

Controlled release spatial repellent devices (CRDs) as novel tools against malaria transmission: a semi-field study in Macha, Zambia.

BACKGROUND: The emergence of mosquitoes that can avoid indoor-deployed interventions, such as treated bed nets and indoor residual spraying, threatens the mainstay of malaria control in Zambia. Furthermore, the requirement for high coverage of these tools poses operational challenges. Spatial repellents are being assessed to supplement these vector control tools, but limitations exist in the residual effect of the repellent and the need for external power or heat for diffusion of the volatiles. METHODS: A semi-field evaluation of a novel controlled release spatial repellent device (CRD) was conducted in Macha, Zambia. These devices emanate metofluthrin with no need for external power. Devices were deployed in huts within the semi-field system (SFS). Female Anopheles gambiae sensu stricto released within the SFS were trapped overnight by light traps and collected by aspiration the next morning inside and outside of huts to determine the extent of mosquito repellency and the impact on host-seeking and survival. Experiments studied the impact of number of devices as well as the presence of hut occupants. The study was complemented with numerical methods based on computational fluid dynamics to simulate spatial distribution of metofluthrin. RESULTS: Presence of CRDs was associated with significant reductions in indoor counts of mosquitoes, regardless of whether huts were occupied or not. Repellency ranged from 15 to 60% compared to huts with no devices. Reducing the number of devices from 16 to 4 had little impact on repellency. When huts were occupied, indoor mosquito host-seeking was higher in the presence of CRDs, whilst survival was significantly reduced. CONCLUSIONS: This study demonstrated that deployment of as few as four CRDs within a hut was associated with reduced indoor mosquito densities. As would be expected, presence of occupants within huts, resulted in greater indoor catches (both with and without devices). The increased indoor mosquito host-seeking and mortality in huts when devices were present may be explained by the excito-repellency activity of metofluthrin. These semi-field experiments provide preliminary data on the utility of CRD spatial repellents to reduce indoor densities of An. gambiae mosquitoes. Studies will further investigate the impact of CRDs on mosquito behaviour as well as epidemiological protective efficacy.

A comparison of the Nzi, Horse Pal® and Bite-Lite® H-traps and selected baits for the collection of adult Tabanidae in Florida and North Carolina.

Despite the veterinary and medical importance of horse flies, deer flies, and yellow flies, only a few trap types have been evaluated to monitor adult population dynamics. Currently, three trap types are being utilized (H-trap, Horse Pal® (HP), and Nzi trap), but no head-to-head comparisons have been reported. Thus, we conducted comparative trapping studies in Florida and North Carolina. At two study sites in Florida, the efficacy of all three trap types was compared, but only the H-trap and HP were compared in North Carolina. Although trap type was significant at all sites, the trap type which caught the most specimens was not the same. In Florida at the Lower Suwannee Wildlife Refuge (LSWR) site, the H-trap caught the most specimens (2,006), followed in decreasing order by Nzi (938) and HP (541). At the Cedar Ridge Ranch site, the Nzi caught significantly more specimens (1,439) than the H-trap (215) and HP (161), which were not significantly different from each other. In North Carolina, the H-trap caught approximately twice as many specimens as the HP (1,458 vs 720). These trap comparison studies were followed up by a study on the efficacy of various bait combinations: (No Bait (NB), dry ice only (DI), Trap Tech Lure (TTL) only, and DI + TTL), which was conducted only at the two Florida sites with H-traps. At both sites, bait combinations significantly affected trap collections. One pattern (DI +TTL > DI > TTL > NB) was recorded at the LSWR, while at the Cedar River Ranch the pattern was DI > DI +TTL > TTL > NB. Our data showed that trap type and bait combination significantly influence overall adult tabanid abundance as well as individual species composition.

Evaluation of Pyriproxyfen Dissemination via Aedes albopictus From a Point-Source Larvicide Application in Northeast Florida.

The Asian tiger mosquito, Aedes albopictus , ranks among the most important vectors of dengue fever, Zika virus, and chikungunya virus. With no specific medications or vaccines available, vector control is the only way to combat these diseases. Autodissemination of the insect growth regulator pyriproxyfen (NyGuard®) from a point-source treatment was evaluated in field settings in northeast Florida. The objective of this study was to investigate the possibility of pyriproxyfen dissemination from a treatment site to nontreated oviposition sites via the skip oviposition behavior of Ae. albopictus. A spray application was made to a tire pile using a Stihl® SR 420 backpack sprayer. Autodissemination oviposition vases containing oak infusion water were positioned in groups of five at 25 to 400 m in 4 transects surrounding the tire pile. Two sets of 5 control vases containing oak infusion water were placed 1,500 m from the tire pile and oak infusion water samples were collected directly from the tire pile. Fifty milliliter samples were extracted from each vase weekly and preserved for pyriproxyfen residue analysis. All vases were analyzed at week 0 (4 h post-treatment), 1, 2, 4, and 6. Overall, there were no differences in pupal mortality between the control and autodissemination vases. The tire pile samples had significantly more mortality (P < 0.0001) out to 4 wk when compared to autodissemination and control vases.

The Knight Stick Trap and Knight Stick Sticky Wraps: New Tools for Stable Fly (Diptera: Muscidae) Management.

Stable fly, Stomoxys calcitrans (L.) (Diptera: Muscidae), management can be difficult, especially in situations where pesticide usage is restricted or disallowed. Traps have been used for monitoring stable flies, but have rarely been used for management. The Knight Stick (KS) trap recently became available, and preliminary studies indicated that it might be an improvement to traps currently in use. The Olson Sticky Fly trap was chosen as the control trap for the purpose of comparisons. Both traps attract stable flies by alteration of light waves and capture flies on a sticky wrap covering the trap base. The KS trap captured 3× more stable flies than the Olson trap, whereas the Olson trap base covered with the KS Sticky wrap captured 3-5× more stable flies than the Olson trap base with the standard Olson Sticky wrap. This indicated inherent attraction from the KS Sticky wrap. This was supported when KS Tank wraps, a larger version of the KS Sticky wraps, applied to 51 kg of liquid propane (LP) tanks on Mosquito Magnet Independence traps producing CO2, captured significantly more stable flies and significantly more stable flies per square centimeter of sticky wrap than Olson Sticky Sleeve wraps applied to the LP tanks. In a final study, when two configurations of KS Tank wraps were applied to white plastic barrels and compared with three standard KS traps, mean numbers of stable flies captured were numerically similar. The significance of findings and potential uses for the traps are discussed.

Evaluations of dual attractant toxic sugar baits for surveillance and control of Aedes aegypti and Aedes albopictus in Florida.

BACKGROUND: Dual attractant toxic sugar baits (D-ATSB) containing two host kairomones, L-lactic (LA) and 1-octen-3-ol (O), and fruit-based attractants were evaluated through olfactory, consumption and mortality, and semi-field experiments to determine if host kairomones could first, enhance attraction of a fruit-based (attractant) toxic sugar bait (ATSB), and second, increase the efficacy of a fruit based attractive toxic sugar bait (ATSB). METHODS: Four combinations of LA and O were incorporated into the ATSB and evaluated in an olfactometer to determine if these combinations could enhance attraction of Aedes aegypti (L.) to the bait. Ae. albopictus (Skuse) and Ae. aegypti were used to determine bait consumption through excrement droplet counts and percent mortality, of the most attractive D-ATSB (1% LA and 1% O) from the olfactory study. Semi-field evaluations were conducted in screened portable field cages to determine if the D-ATSB applied to non-flowering plants controlled more mosquitoes than the fruit-based ATSB, and ASB. Mosquitoes were exposed to D-ATSB and the two controls for 48 h and collected with BGS traps. The catch rates of the BGS traps were compared to determine efficacy of the D-ATSB. RESULTS: During olfactometer evaluations of D-ATSB, Ae. aegypti mosquitoes were more attracted to 1% LA and 1% O compared to the fruit-based toxic sugar bait alone. Both species of mosquito consumed more fruit-based non-toxic bait (ASB) and ATSB than the D-ATSB. For both species, percent mortality bioassays indicated D-ATSB controlled mosquitoes, as compared to non-toxic control, but not more than the fruit based ATSB. Semi-field evaluations, BioGents sentinel traps at 48 h confirmed that ATSB (positive control) controlled Ae. albopictus, but there was no statistical difference between ASB (negative control) and the D-ATSB. No differences were observed between the mosquitoes caught in any of the experimental formulations for Ae. aegypti. CONCLUSIONS: L-lactic (1%) and 1-octen-3-ol (1%) added to a fruit-based sugar bait increased attraction of Ae. aegypti and may have future implications in mosquito trapping devices. The addition of the host kairomones did not enhance the consumption and efficacy of the ATSB in laboratory or semi-field evaluations for both mosquito species. We attribute to the absence of other host cues leading to lack of alighting onto bait surfaces to imbibe the toxic bait, as well as a possible decrease in palatability of the bait caused by the addition of the host kairomones.

Assessment of Aedes albopictus (Skuse) (Diptera: Culicidae) clutch size in wild and laboratory populations.

Aedes albopictus (Skuse) is an invasive mosquito species found across the southern U.S. with range expansion into many northern states. Intra- and interspecific larval competition have been evaluated for Ae. albopictus with respect to subsequent adult size, immature and adult survivability, and its capacity to vector pathogens as an adult. However, limited data are available on egg production as related to larval rearing conditions. Because Ae. albopictus is a container-inhabiting mosquito that oviposits in resource-limited habitats, it is found under variable density-dependent conditions. Therefore, we examined the impact of specific rearing conditions on Ae. albopictus clutch size and adult body size; comparing the egg production values and wing lengths from known developmental densities to those from field-collected populations. Field populations varied significantly among collection sites in mean clutch size (23 to 46). These clutch sizes were comparable to the mean clutch sizes of females reared at the larval densities of nine (20 eggs) and three (53 eggs) larvae per 3 ml of water in the laboratory. Field populations experienced density-dependent effects impacting adult mosquito size. Mosquitoes from the four sample sites had mean wing lengths of 1.99, 2.47, 2.51, and 2.54 mm, which were less than the mean wing length of mosquitoes reared at larval densities of three larvae per 3 ml of water (2.57 mm).

Aedes albopictus (Diptera: Culicidae) Oviposition Preference as Influenced by Container Size and Buddleja davidii Plants.

Aedes albopictus (Skuse) is a container-breeding mosquito commonly found in residential areas of its range in the United States. Mosquitoes are known to utilize flowering plants for sugar acquisition. Limited information is known about the influences on oviposition site selection, outside of container size. Residential areas are often landscaped with a variety of flowering plants and are known to provide numerous sizes of potential larval developmental sites for container-breeding mosqutioes. Through screened enclosure and field studies, the oviposition preference of Ae. albopictus for containers of three selected sizes (473, 946 and 1,892 ml) and the influence of flowering butterfly bush (Buddleja davidii Franchett cultivar 'Guinevere') plants were examined. Our results document that significantly more eggs were oviposited in the largest containers. Additionally, significantly more eggs were oviposited in containers adjacent to flowering butterfly bushes than in those without a flowering butterfly bush. Finally, our results document that flowering butterfly bushes exerted greater influence over Ae. albopictus oviposition decisions than did container size. Our findings can be applied to several aspects of Ae. albopictus surveillance and control.

Development and Evaluation of an Attractive Self-Marking Ovitrap to Measure Dispersal and Determine Skip Oviposition in Aedes albopictus (Diptera: Culicidae) Field Populations.

Aedes albopictus (Skuse) is a container-breeding species with considerable public health importance. To date, Ae. albopictus oviposition behavior has been assessed in outdoor conditions, but only with laboratory-reared specimens. In outdoor large-cage and field studies, we used an attractive self-marking ovipositional device to assess Ae. albopictus skip oviposition behavior. In field studies, 37 wild Ae. albopictus that visited an attractive self-marking ovisite were subsequently captured at a sticky ovitrap within a 4-d period. Because the average Ae. albopictus gonotrophic period is 4.5-6 d, the wild-caught Ae. albopictus visited at least two oviposition sites within a single gonotrophic period. This provided field-based indirect evidence of skip oviposition. The mean distance traveled (MDT) during the 20-d evaluations ranged from 58 to 78 m. The maximum observed distance traveled was 149 m, which was the outer edge of our trapping ability. As populations of Ae. albopictus increased, the MDT during the 4- and 20-d post-marking period increased significantly. Additional observations of wild-marked and captured Aedes triseriatus (Say) are discussed.

The Effects of Larval Habitat Quality on Aedes albopictus Skip Oviposition.

Aedes albopictus is an invasive mosquito species that transmits human-disease-causing pathogens. It is a container-inhabiting species that oviposits in resource-limited habitats. To mitigate larval competition, Ae. albopictus females may choose to distribute eggs from a single gonotrophic cycle among multiple containers through skip oviposition. With the use of individual females released in indoor and outdoor caged trials, we evaluated the oviposition choices made by gravid Ae. albopictus offered larval habitats with different qualities. Our results demonstrate that Ae. albopictus performs skip oviposition and that the degree of egg distribution is related to the quality of the larval habitat. In a 4-choice arena, individual Ae. albopictus oviposited in fewer containers when presented with ovisites of high-quality larval habitat (uncrowded conditions) compared with oviposition in low-quality (crowded conditions) larval habitats. Additionally, the females selectively oviposited in high-quality habitats when offered both low- and high-quality habitats, but distributed eggs more evenly among multiple high-quality habitats. Our results have important implications for mosquito management plans that include the use of lethal ovitraps, as well as the role of this behavior in distribution of disease-causing pathogens.

A Comparison of Carbon Dioxide Sources for Mosquito Capture in Centers for Disease Control and Prevention Light Traps on the Florida Gulf Coast (1).

Traditional sources of carbon dioxide (CO2), dry ice, and compressed gas, were tested against 3 combinations of food-grade reagents known to generate CO2using a compact, lightweight generator delivery system with Centers for Disease Control and Prevention light traps. Three 6 × 6 Latin square trials were completed near the Florida Gulf Coast in the Lower Suwannee Wildlife Refuge during the summer of 2013, collecting a total of 31,632 female mosquitoes. Treatments included dry ice, compressed CO2gas, a control trap (no CO2), citric acid + sodium bicarbonate, vinegar + sodium bicarbonate, and yeast + sugar. Decreasing order of trap collections (treatment mean number of mosquitoes per trap night ± standard error) were dry ice 773.5 (± 110.1) > compressed gas 440.7 (± 42.3) > citric acid + sodium bicarbonate 197.6 (± 30.4), yeast + sugar 153.6 (± 27.4) > vinegar + sodium bicarbonate 109.6 (± 16.2) > control 82.4 (± 14.0). A 2-way Kruskal-Wallis analysis by treatment, site, and treatment × site interaction identified significant differences between all treatments. Although dry ice and compressed CO2gas collected significantly more mosquitoes than other combinations (P < 0.05), use of citric acid and sodium bicarbonate or yeast and sugar greatly outperformed unbaited traps and offer a good alternative to dry ice and compressed gas in areas where these agents are not readily available or are difficult to obtain due to logistical constraints. An inexpensive, portable CO2generator for use with food-grade reagents is described.

Control of sand flies with attractive toxic sugar baits (ATSB) and potential impact on non-target organisms in Morocco.

BACKGROUND: The persistence and geographical expansion of leishmaniasis is a major public health problem that requires the development of effective integrated vector management strategies for sand fly control. Moreover, these strategies must be economically and environmentally sustainable approaches that can be modified based on the current knowledge of sand fly vector behavior. The efficacy of using attractive toxic sugar baits (ATSB) for sand fly control and the potential impacts of ATSB on non-target organisms in Morocco was investigated. METHODS: Sand fly field experiments were conducted in an agricultural area along the flood plain of the Ourika River. Six study sites (600 m x 600 m); three with "sugar rich" (with cactus hedges bearing countless ripe fruits) environments and three with "sugar poor" (green vegetation only suitable for plant tissue feeding) environments were selected to evaluate ATSB, containing the toxin, dinotefuran. ATSB applications were made either with bait stations or sprayed on non-flowering vegetation. Control sites were established in both sugar rich and sugar poor environments. Field studies evaluating feeding on vegetation treated with attractive (non-toxic) sugar baits (ASB) by non-target arthropods were conducted at both sites with red stained ASB applied to non-flowering vegetation, flowering vegetation, or on bait stations. RESULTS: At both the sites, a single application of ATSB either applied to vegetation or bait stations significantly reduced densities of both female and male sand flies (Phlebotomus papatasi and P. sergenti) for the five-week trial period. Sand fly populations were reduced by 82.8% and 76.9% at sugar poor sites having ATSB applied to vegetation or presented as a bait station, respectively and by 78.7% and 83.2%, respectively at sugar rich sites. The potential impact of ATSB on non-targets, if applied on green non-flowering vegetation and bait stations, was low for all non-target groups as only 1% and 0.7% were stained with non-toxic bait respectively when monitored after 24 hours. CONCLUSIONS: The results of this field study demonstrate ATSB effectively controls both female and male sand flies regardless of competing sugar sources. Furthermore, ATSB applied to foliar vegetation and on bait stations has low non-target impact.