Large-scale production of mosquitoes is a key factor for a successful sterile insect technique program. A manageable mass-production cage must contain appropriate features for adult resting, mating, feeding, and ovipositional activities. In order to maximize egg collections, tests were conducted to determine the physical characteristics of ovipositional sites for caged Anopheles arabiensis. Effects of texture, shade, height, and shape of the ovipositional container on female behavior were investigated. Results indicate a strong preference for oviposition on humid substrates over free-standing water. The shade and texture of the cup's walls also influenced site choice, with black rough inner vertical walls of the cup resulting in the largest number of eggs. Ovipositional sites with square shape were preferred rather than circular cups, and in the square cups, >60% of the eggs were laid in the corners. Height also affected oviposition, as An. arabiensis significantly favored the lowest sites even though some oviposition occurred at higher sites. Based on this study and from available literature, we determined the characteristics of an artificial ovipositional site that includes all these characteristics, which will yield large numbers of eggs required for mass production of this species.

Moisture plays a major role in the dynamics of mosquito populations, especially those breeding in container habitats. Despite this importance, the role of moisture conditions as they affect oviposition and egg development in Aedes vectors remains largely unexplored. We investigated the effect of exposing gravid female Aedes albopictus mosquitoes and their eggs to different moisture levels (MLs) for various periods on oviposition and hatching. Overall, high-moisture substrates (HMSs; 66% and 72%) provided better environments for egg laying. The timing of initial egg laying was far longer at the lowest substrate moisture level (LSML, 25% and 41.2%) than at HMSs. The numbers of eggs laid were much lower in the drier environments. At LSMLs, gravid females retained increasing numbers of mature eggs until death, and egg retention decreased gradually with increasing ML. The HMSs also provided better environments for larval eclosion. The numbers of eggs hatched were lower at the LSML than the HSML environment. No egg hatching occurred after 1 h exposure to moisture. However, egg hatching occurred by installment, with spontaneous hatching (SH) increasing gradually with increasing ML. High-moisture conditions combined with long exposure (30 h and 48 h) favored SH. These results suggest that Ae. albopictus females can respond to better moisture conditions for increased success of embryonation and larval eclosion. This information may be useful in the colonization of floodwater Aedes species.

We describe here an in vitro behavioral assay for testing mosquito repellents applied in a dose-based manner to a warm body (34°C) in test cages. The system was used to assess the sensitivity of 4–6-day-old Anopheles gambiae to the insect repellent diethyl methyl benzamide (deet). These tests were made in the absence and presence of additional carbon dioxide (CO₂) applied as a pulse to activate mosquitoes in the cages. In the absence of the CO₂ pulse the mosquitoes hardly responded to the warm body. Increasing the CO₂ level in the cage by 1,000 parts per million caused a 25-fold increase in the number of landings by mosquitoes on the warm body in 2-min tests. This mosquito activation allowed the measurement of a significant reduction in the number of landings to bite on the warm body with increasing doses of deet (0.4 to 3.8 µg/cm²). An asymptotic nonlinear model fitted to the repellency data in the presence of CO₂ allowed estimation of the effective dose of deet that reduced landings to bite by 50% (ED₅₀) at 0.95 µg/cm² (5 nmol/cm²) and the corresponding ED₉₅ at 4.12 µg/cm² (21.5 nmol/cm²). This in vitro bioassay has the advantage of permitting a fast throughput of test products under standardized conditions and is suitable for screenings designed for the purpose of discovering lead products with as yet unknown human toxicological and dermatological profiles.

The efficacy of alpha-cypermethrin (Cyperthor®) and lambda-cyhalothrin (Demand®) to prevent mosquito larval colonization of water-containing receptacles was investigated using 2 differing applications in disused car tires in Darwin, Australia. Insecticide treatments were applied uniformly to the inside surfaces of 2 categories of tires: 1) dry tires that were partially filled with water 24 h after spraying and 2) wet tires partially filled with water prior to spraying. All mosquito larvae, pupae, and dead adults were collected from the treatment and control tires weekly over the 24-wk study period and were later identified to species in the laboratory. Control tires were colonized by Aedes notoscriptus in wk 2 and by Culex quinquefasciatus in wk 4. Aedes notoscriptus failed to colonize any alpha-cypermethrin–treated tires until wk 22 and did not colonize any lambda-cyhalothrin–treated tires during the 24-wk trial. Culex quinquefasciatus colonized alpha-cypermethrin– and lambda-cyhalothrin–treated tires from wk 11 and wk 15, respectively. These results indicate both insecticides using either application method can prevent colonization of Ae. notoscriptus for at least 20 wk and demonstrate great potential for the prevention of breeding in receptacles for other receptacle-breeding Aedes species, such as the dengue vectors, Ae. aegypti and Ae. albopictus.

The purpose of this study was to evaluate the use of boric acid, imidacloprid, ivermectin, or abamectin incorporated into sugar baits as oral toxicants for adult phlebotomine sand flies. Variable toxicity of insecticide–sugar bait solutions to adult male and female sand flies was demonstrated, based on male–female median lethal concentration values of 0.10–0.08, 6.13–9.53, and 9.03–18.11 mg/liter of imidacloprid, ivermectin, and abamectin, respectively. Complete control of sand flies could not be achieved with as high as 40 g/liter of boric acid in sugar bait solution; concentrations >40 g/liter were found repellent to the sand flies. Uranine O (a fluorescent tracer dye that can be used to measure the ingestion of sugar baits by sand flies) did not interact negatively with imidacloprid, ivermectin, or abamectin when it was combined with the insecticides in a sugar bait. Also, incorporation of imidacloprid, ivermectin, or abamectin into sugar baits did not reduce the effect whether adult male and female sand flies fed on these sugar baits. We propose that imidacloprid, ivermectin, or abamectin could be used to control adult sand fly populations with targeted use of insecticide-treated sugar baits.

Aedes aegypti larvae from 8 sites in Trinidad and 1 in Tobago were assayed against temephos, malathion, and fenthion using the Centers for Disease Control and Prevention time-mortality–based bioassay method. Resistance ratios (RRs) and resistance thresholds (RTs) for each insecticide were calculated in relation to the Caribbean Epidemiology Center reference susceptible strain. Results showed that the Haleland Park and Tobago strains were susceptible to fenthion and malathion, respectively (RRs < 1), while the San Fernando strain had a high RR (33.92) to malathion. All other strains had low-level resistance to fenthion and malathion. Resistance to temephos was more intense with 4 strains showing high-level resistance. The established RT was 60 min for fenthion, 75 min for bendiocarb, and 120 min for temephos and malathion. At the RTs, all Trinidad strains were resistant to temephos (11.50–74.50% mortality), 7 resistant to fenthion (21.25–78.75% mortality), and 5 resistant to malathion (56.25–77.50% mortality). The other strains were incipiently resistant (80–97% mortality). Despite the discrepancies between the RR levels and RT status, it is evident that the organophosphate insecticide resistance is prevalent in Trinidad and Tobago populations of Ae. aegypti. These results suggest that operational failure could soon occur and alternative strategies should be developed and implemented to reduce the probability of further selection pressure on resistant Ae. aegypti populations in Trinidad and Tobago.

The use of bioassay cages in the efficacy assessment of pesticides, application techniques, and technologies is common practice using numerous cage designs, which vary in both shape and size as well as type of mesh. The objective of this work was to examine various cage shapes and mesh types for their filtration effects on air speed, spray droplet size, and spray volume. Reductions in wind speed and droplet size seen inside the cages were measured by placing cages in a low-speed wind tunnel at air speeds of 0.5 m/sec, 1 m/sec, 2 m/sec, and 4 m/sec and cage face orientations (relative to the air stream) of 0°, 10°, 22.5°, and 45°. Reduction in spray volume inside a select number of cages was also evaluated under similar conditions. Generally, greater air speed reductions were seen at lower external air speeds with overall reductions ranging from 30% to 88%, depending on cage type and tunnel air speed. Cages constructed with screens of lower porosities and smaller cylindrical-shaped cages tended to provide greater resistance to air flow and spray volume. Overall, spray droplet size inside the cages was minimally reduced by 0–10%. There was a 32–100% reduction in concentration of the spray volume applied relative to that recovered inside the bioassay cages, depending on the cage geometry and screening material used. In general, concentration reductions were greatest at lower air speeds and for cages with lower porosity screens. As a result of this work, field researchers involved in assessing the efficacy of vector control applications will have a better understanding of the air speed and spray volume entering insect bioassay cages, relative to the amount applied, resulting in better recommended application techniques and dosage levels.

A study was conducted to analyze the performance of 3 electrostatic (Electrolon BP-2.5™, Spectrum Electrostatic 4010, and Spectrum Electrostatic head on a Stihl 420) and 2 conventional (Buffalo Turbine CSM2 and Stihl 420) sprayers for barrier sprays to suppress an adult mosquito population in an enclosed area. Sprayer characteristics such as charge-mass ratio, air velocity, flow rate, and droplet spectra were measured while spraying water. Dispersion of the spray cloud from these sprayers was determined using coverage on water-sensitive cards at various heights (0.5 m, 1.0 m, 1.5 m, 2.0 m, 2.5 m, and 3.0 m) and depths (1 m, 3 m, and 5 m) into the under-forest vegetation while spraying bifenthrin (Talstar™ 7.9% AI; FMC Corporation, Philadelphia, PA) at the rate of 21.8 ml/300 m of treated row. The charge-mass ratio data show that Electrostatic head on a Stihl 420 did not impart enough charge to the droplets to be considered as an electrostatic sprayer. In general, the charged spray cloud moved down toward the ground. The Electrolon BP 2.5 had significantly lower spray coverage on cards, indicating lack of spray dispersion. This sprayer had the lowest air velocity and did not have the air capacity needed to deliver droplets close to the target for electrostatic force to affect deposition. The analysis shows that these 2 sprayers are not a suitable choice for barrier sprays on vegetation. The results indicate that the Buffalo Turbine is suitable for barriers wider than 3 m, and the Spectrum 4010 and Stihl 420 are suitable for 1–3-m–wide barriers.

The malarial parasite, Plasmodium vivax, was detected in 4 species of Anopheles (Hyrcanus Group) mosquitoes, namely An. kleini, An. pullus, An. belenrae, and An. sinensis, from Gyeonggi Province, Republic of Korea (ROK). This study confirmed that An. belenrae was infected by P. vivax, and implicated this species as a potential vector of vivax malaria in the ROK.

The larvicidal efficacy of the acetone extract of the thalli of Caulerpa scalpelliformis var. denticulata was determined against late 2nd to early 3rd instars of Culex pipiens at concentrations ranging from 100 to 2,000 parts per million (ppm). At 1,200 ppm, the extract caused >70% larval mortality at 24-, 48-, and 72-h exposure. The LC₅₀ (lethal concentration) and LC₉₀ values of C. scalpelliformis were 338.91 and 1,891.31 ppm, respectively. Our data showed that this species of seaweed contains components with larvicidal properties against mosquitoes.

Tissue cultures are used to propagate viruses for use in mosquito infection studies and to detect live virus in field-collected specimens. Microscopic evaluation of cytopathic effects is used to visualize virus presence. In this study, both low-passage (LP; n  =  35) and high-passage (HP; n  =  218) lines of African green monkey kidney (Vero) cells were infected with West Nile virus, and virus growth kinetics were quantitated over different incubation periods (IPs) (2, 6, 10, 24, 48, 72, and 96 h postinfection). Virus titers were significantly (P < 0.05) higher in the HP compared with LP line 24, 48, and 96 h postinfection. No differences were observed in plaque morphology between the LP and HP lines. Culex pipiens quinquefasciatus were fed infectious blood meals created using supernatant from the 2 lines and maintained at 28°C for a 12-day IP. Although the virus dose was higher (P < 0.05) in the HP compared with the LP line, there were no significant differences in mosquito infection or dissemination rates at the end of the IP. The significance of these observations for assessing virus presence and pathogenicity is discussed.

The phlebotomine sand fly, Lutzomyia shannoni (Dyar), is considered both zoophillic and anthropophillic and may be a potential vector of human and livestock pathogens. While the known range of this species in North America has recently been expanded, many aspects of its natural history, which are fundamental components for assessments of disease risk or development of integrated pest management programs for the vector, remain unclear. One is the time of day when individuals are active and searching for blood meals. Through the use of a rotating trap mechanism, male L. shannoni were found to be active earlier in the evening (1–4 h after sunset), whereas the majority of female activity occurred after midnight.

Rift Valley fever (RVF), a disease of ruminants and humans, has been responsible for large outbreaks in Africa that have resulted in hundreds of thousands of human infections and major economic disruption due to loss of livestock and to trade restrictions. As indicated by the rapid spread of West Nile viral activity across North America since its discovery in 1999 and the rapid and widespread movement of chikungunya virus from Africa throughout the Indian Ocean Islands to Asia and Europe, an introduced exotic arbovirus can be rapidly and widely established across wide geographical regions. Although RVF virus (RVFV) is normally transmitted by mosquitoes, we wanted to determine the potential for this virus to replicate in 2 of the most globally distributed and common higher flies: house flies, Musca domestica, and stable flies, Stomoxys calcitrans. Neither species supported the replication of RVFV, even after intrathoracic inoculation. However, S. calcitrans was able to mechanically transmit RVFV to susceptible hamsters (Mesocricetus auratus) after probing on infected hamsters with high viral titers. Therefore, S. calcitrans, because of its close association with domestic animals that serve as amplifying hosts of RVFV, should be considered a possible mechanical vector of RVFV, and it may contribute to the rapid spread of a RVF outbreak. Other Stomoxys species present in Africa and elsewhere may also play similar roles.

The effectiveness of the attractants l-octen-3-ol (octenol) and l-lactic acid (Lurex™) on the collection of Aedes albopictus, Culex tritaeniorhynchus, Cx. pipiens pallens, and Anopheles sinensis was first evaluated in Mosquito Magnet® Pro traps in Yamenkou and Badachu residential areas, Beijing City, and Lishui area, Zhejiang Province, China. The Mosquito Magnet Pro traps baited with octenol collected significantly more Ae. albopictus, Cx. tritaeniorhynchus, and An. sinensis, but fewer Cx. pipiens pallens than collection by the traps alone. There were no significant differences in the numbers of Cx. pipiens pallens, Cx. tritaeniorhynchus, and An. sinensis collected by Mosquito Magnet Pro traps baited with Lurex compared to the traps alone, but the Mosquito Magnet Pro traps baited with Lurex collected significantly more Ae. albopictus than the number collected by the traps alone at 2 areas in Beijing.