We evaluated the ability of the Rapid Analyte Measurement Platform (RAMP^®) mosquito-grinding buffer to inactivate West Nile virus (WNV) by subjecting WNV-positive samples ground in RAMP buffer to incubation intervals ranging from 5 min to 60 min. At each time point an aliquot was removed and serially diluted in bovine albumin (BA)-1 cell culture media to stop the inactivation process by RAMP buffer. Each BA-1 sample was tested for viable virus using Vero 6-well cell culture plaque assay and observed for plaques. We observed very limited inactivation of WNV (1–2 log₁₀ plaque-forming units/ml) by RAMP buffer. Concerned for RAMP operators who may be using this assay in low-level biocontainment facilities, we developed an alternate sample homogenization protocol using Triton X-100 detergent that ensures complete WNV inactivation without compromising the performance of the RAMP assay.

An adult mosquito survey was conducted at 12 sites using carbon dioxide traps in northern Minnesota throughout the summer of 2012. Specimens were counted, identified to species, sorted into pools, and tested for eastern equine encephalitis (EEEV) and West Nile virus (WNV). Our findings extend the known range of Culiseta melanura, Anopheles barberi, and An. quadrimaculatus and document the presence and abundance of 27 other mosquito taxa in the region. None of the pools tested positive for EEEV or WNV.

Gravid mosquito collections were compared among several large-volume (infusion volume ≥35 liters) gravid trap designs and the small-volume (infusion volume  =  6 liters) Centers for Disease Control and Prevention (CDC) gravid trap used routinely by vector control districts for vector and pathogen surveillance. The numbers of gravid Culex quinquefasciatus, Cx. tarsalis, and Cx. stigmatosoma collected by large gravid traps were greater than by the CDC gravid trap during nearly all overnight trials. Large-volume gravid traps collected on average 6.6-fold more adult female Culex mosquitoes compared to small-volume CDC gravid traps across 3 seasons during the 3 years of the studies. The differences in gravid mosquito collections between large-versus small-volume gravid traps were greatest during spring, when 8- to 56-fold more Culex individuals were collected using large-volume gravid traps. The proportion of gravid females in collections did not differ appreciably among the more effective trap designs tested. Important determinants of gravid trap performance were infusion container size and type as well as infusion volume, which determined the distance between the suction trap and the infusion surface. Of lesser importance for gravid trap performance were the number of suction traps, method of suction trap mounting, and infusion concentration. Fermentation of infusions between 1 and 4 wk weakly affected total mosquito collections, with Cx. stigmatosoma collections moderately enhanced by comparatively young and organically enriched infusions. A suction trap mounted above 100 liters of organic infusion housed in a 121-liter black plastic container collected the most gravid mosquitoes over the greatest range of experimental conditions, and a 35-liter infusion with side-mounted suction traps was a promising lesser-volume alternative design.

Phlebotominae are the vectors of Leishmania parasites. It is important to have available surveillance and collection methods for the sand fly vectors. The objectives of the present study were to evaluate and compare traps for the collection of sand fly species and to analyze trap catches along months and transects. Field evaluations over a year were conducted in an endemic area of leishmaniasis in the state of Quintana Roo, Mexico. A randomized-block design was implemented in study area with tropical rainforest vegetation. The study design utilized 4 transects with 11 trap types: 1) Centers for Disease Control and Prevention (CDC) light trap with incandescent bulb (CDC-I), 2) CDC light trap with blue light-emitting diodes (LEDs) (CDC-B), 3) CDC light trap with white LEDs (CDC-W), 4) CDC light trap with red LEDs (CDC-R), 5) CDC light trap with green LEDs (CDC-G), 6) Disney trap, 7) Disney trap with white LEDs, 8) sticky panels, 9) sticky panels with white LEDs, 10) delta-like trap, and 11) delta-like trap with white LEDs. A total of 1,014 specimens of 13 species and 2 genera (Lutzomyia and Brumptomyia) were collected. There were significant differences in the mean number of sand flies caught with the 11 traps; CDC-I was (P  =  0.0000) more effective than the other traps. Other traps exhibited the following results: CDC-W (17.46%), CDC-B (15.68%), CDC-G (14.89%), and CDC-R (14.30%). The relative abundance of different species varied according to trap types used, and the CDC-I trap attracted more specimens of the known vectors of Leishmania spp., such as like Lutzomyia cruciata, Lu. shannoni, and Lu. ovallesi. Disney trap captured more specimens of Lu. olmeca olmeca. Based on abundance and number of species, CDC light traps and Disney traps appeared to be good candidates for use in vector surveillance programs in this endemic area of Mexico.

Based on an ethnobotanical study on use for plant species against mosquito bites in the Kota Tinggi District, Johor State, Malaysia, 3 plants selected for study, Citrus aurantifolia (leaves), Citrus grandis (fruit peel), and Alpinia galanga (rhizome), were extracted using hydrodistillation to produce essential oils. These essential oils were then formulated as a lotion using a microencapsulation process and then tested for their repellent effect against Aedes aegypti. N,N-diethyl-m-toluamide (deet) was also prepared in the same formulation and tested for repellency as controls. Four commercial plant-based repellent (KAPS^®, MozAway^®, BioZ Natural^®, and Mosiquard^®) also were incorporated in the bioassay for comparison purposes. Bioassays revealed that at 20% concentration all repellent formulations demonstrated complete protection for 2 h and >90% for 4 h post-application. The A. galanga–based formulation provided the greatest level of protection (98.91%), which extended for 4 h post-application and was not significantly different from deet at similar concentration. When compared with commercial plant-based repellents (KAPS^®, MozAway^®, and BioZ Natural^®), the 3 lotion formulations showed significantly better protection against Ae. aegypti bites, providing >90% protection for 4 h. In conclusion, our 3 plant-based lotion formulations provided acceptable levels of protection against host-seeking Ae. aegypti and should be developed.

Malaria control methods rely mostly on adult mosquito control using insecticide-treated nets and indoor residual spraying with insecticides. Plants such as endod (Phytolacca dodecandra) can potentially be used for the control of mosquito larvae as a supplement to adult control methods. Following the discovery of endod, a molluscicide plant, more than 5 decades ago in Ethiopia, subsequent studies have shown that its potency can further be increased by simple procedures such as aging endod berry powder in water. This study was conducted to evaluate the killing effect of fresh and aged endod solution against 4th-stage larvae of Anopheles arabiensis. Laboratory-reared An. arabiensis larvae exposed to different concentrations of endod preparation using distilled or spring water had 50% lethal concentration (LC₅₀)  =  49.6 ppm and 90% lethal concentration (LC₉₀)  =  234 ppm for fresh and LC₅₀  =  36.4 ppm and LC₉₀  =  115.7 ppm for the aged endod solution in distilled water against the laboratory population. Against field-collected larvae of the same species, aged preparations in habitat water resulted in higher LC₅₀ (472.7 ppm) and LC₉₀ (691 ppm) values, with only a slight improvement over fresh preparations in habitat water (LC₅₀  =  456.2 ppm; LC₉₀  =  896.1 ppm). In general, although aged preparations of endod required lower concentrations than fresh to kill at least 90% of the larvae, these concentrations were much higher (12–70×) than that required for schistosome-transmitting snails.

The efficacy of a metofluthrin-impregnated net (MIN) known as the “Mushikonazu” on the house entry behavior of female Aedes aegypti and Anopheles dirus mosquitoes was evaluated using a semi-field 50-m tunnel setup. While the MIN is labeled for the control of chironomids and moth flies, this study determined the feasibility of using the device, given its current construction and metofluthrin formulation, as a spatial repellent against mosquitoes. Sentinel and cone bioassays were used to determine the insecticidal effect of the MIN. A spatial activity index (SAI) was calculated to evaluate responses of the mosquitoes. For the spatial repellent evaluation against Ae. aegypti, the overall mean of SAI was slightly less than 0 at wk 1 after the MIN application and then decreased for the last 4 wk showing a preference to treatment tent. For An. dirus, the mean SAI at wk 1 was positive, indicating a presumed repellent effect of the MIN against An. dirus. For the subsequent 4 wk, the SAI was negative, indicating a preference for the MIN. Results suggested that the MIN may not be a promising approach to repel Ae. aegypti and An. dirus under field conditions in Thailand. However, it remains probable that the MIN may be effective as a spatial repellent if modifications are made to the metofluthrin concentration or formulation and/or the construction of the device.

Ambient insecticides are receiving increasing attention in many developed countries because of their value in reducing mosquito nuisance. As required by the European Union Biocidal Products Regulation 528/2012, these devices require appropriate testing of their efficacy, which is based on estimating the knockdown and mortality rates of free-flying (free) mosquitoes in a test room. However, evaluations using free mosquitoes present many complexities. The performances of 6 alternative methods with mosquitoes held in 2 different cage designs (steel wire and gauze/plastic) with and without an operating fan for air circulation were monitored in a test room through a closed-circuit television system and were compared with the currently recommended method using free mosquitoes. Results for caged mosquitoes without a fan showed a clearly delayed knockdown effect, whereas outcomes for caged mosquitoes with a fan recorded higher mortality at 24 h, compared to free mosquitoes. Among the 6 methods, cages made of gauze and plastic operating with fan wind speed at 2.5–2.8 m/sec was the only method without a significant difference in results for free mosquitoes, and therefore appears as the best alternative to assess knockdown by ambient insecticides accurately.

Widespread resistance of insect pests to insecticides has been widely reported in China and there is consequently an urgent need to adjust pest management strategies appropriately. This requires detailed information on the extent and causes of resistance. The aim of the present study was to investigate levels of resistance to 5 insecticides among 12 strains of Culex tritaeniorhynchus, a major vector of Japanese encephalitis in China. Resistance to deltamethrin, beta-cypermethrin, permethrin, dichlorvos, and propoxur were measured using larval bioassays. The allelic frequency of knockdown resistance (kdr) and acetylcholinesterase (AChE) mutations were determined in all strains. Larval bioassay results indicated that the field strains collected from different sites were resistant to deltamethrin, beta-cypermethrin, permethrin, dichlorvos, and propoxur, with resistance ratio values ranging from 1.70- to 71.98-fold, 7.83- to 43.07-fold, 3.54- to 40.03-fold, 291.85- to 530.89-fold, and 51.32- to 108.83-fold, respectively. A polymerase chain reaction amplification of specific alleles method for individual was developed to detect genotypes of the AChE gene mutation F455W in Cx. tritaeniorhynchus. The frequency of the AChE gene mutation F455W was 100.00% in all strains, making this mutation of no value as a marker of resistance to organophosphorous and carbamate pesticides in Cx. tritaeniorhynchus in China. The kdr allele was present in all strains at frequencies of 10.00–29.55%. Regression analysis indicated a significant correlation between kdr allele frequencies and levels of resistance to deltamethrin, beta-cypermethrin, and permethrin. These results highlight the need to monitor and map insecticide resistance in Cx. tritaeniorhynchus and to adjust pesticide use to minimize the development of resistance in these mosquitoes.

The larvae of the genus Toxorhynchites are typically found in sylvatic habitat environments. They have been proposed as potential biocontrol agents against disease vectors such as Aedes, specifically the larvae of Ae. aegypti and Ae. albopictus. The only 2 species of Toxorhynchites identified in Taiwan are Tx. aurifluus and Tx. manicatus, but there have been no published records of either species since 1984. In January 2014, we identified 2 4th-stage Tx. aurifluus larvae and 2 4th-stage Tx. manicatus larvae in artificial habitats close to dwellings in Guichong village, Pingtung County, Taiwan. Here we update the bionomics of the larvae of both species, including coexisting aquatic fauna, the physicochemical parameters of the larval habitats, and developmental time. We further compare the findings with past records.

Culex pipiens and Cx. restuans are the main vectors of West Nile virus and the primary target species of surveillance and control programs in Pennsylvania. Performing adult control, specifically ultra-low volume (ULV) applications, at night during peak oviposition activity time(s) is necessary to control these species. In July and August of 2009, collections were made at 15-min intervals starting at sunset and continuing until 3 h after sunset to establish a more accurate timeline of Cx. pipiens and Cx. restuans oviposition flight activity. The highest numbers of Cx. pipiens and Cx. restuans were collected during the 15–30, 30–45, and 45–60 min postsunset time intervals (P < 0.05). Oviposition activity began to decrease after 60 min postsunset. These observations have identified a smaller oviposition activity period for Cx. pipiens and Cx. restuans than noted from other studies, thus potentially improving the timing of ULV operations to control these 2 vector species.

Aedes albopictus is an invasive species that poses a health threat in many residential neighborhoods throughout Florida. Aedes albopictus is a high priority for mosquito control efforts in the state. The efficacy of DeltaGard® (AI 2% deltamethrin) application against Ae. albopictus was evaluated in a residential area in St. Augustine, FL. DeltaGard was applied using a truck-mounted ultra-low-volume aerosol generator along 3 streets in a residential neighborhood. Caged mosquito mortality and droplet density data were recorded. Leaf clippings from houses on treated streets were bioassayed against laboratory-reared Ae. albopictus. Overall, the DeltaGard application was found to be more effective in the front yard of the houses, resulting in 78.3% mortality in caged mosquitoes, 42 % mortality in leaf bioassays, and 50.5 nl/cc in spray density. Based on the amount of vegetation and residential barriers around the houses, the application caused only 46.3% mortality in caged mosquitoes, 7.5% mortality in leaf bioassays, and 5.4 nl/cc in spray density in the back yard sites.

The Anastasia Mosquito Control District (AMCD) tests all equipment before field use to determine if machines are suitable for the needs of the district. Three handheld ultra-low-volume (ULV) foggers—the American LongRay (ULV) Fogger Model 3600B with rechargeable lithium battery (DC model), American LongRay ULV Fogger Model 3600E with 110V or 220V AC power (AC model), and Boston Fog Battery Motorized Fogger (Boston Fogger)—were compared to determine which fogger would be most suitable for use by AMCD. Mortality of caged Aedes albopictus was analyzed after 24 h to determine the success of a single application. All 3 foggers resulted in 100% mortality after 24 h using the insecticide Aqualuer 20-20 (active ingredients permethrin 20.6% and piperonyl butoxide 20.6%) 1:5 dilution with reverse osmosis water. Based on operator safety, robustness, and operational performance, the American LongRay DC model was found to be the most suitable at administering Aqualuer 20-20 against caged adult Ae. albopictus.

Testing of the PSO BASDKA-AC1200 multifunction ultrafine particle atomization machine, a thermal fog machine, with Aqualuer 20-20^® (permethrin 20.6%, piperonyl butoxide 20.6%) was conducted against Aedes albopictus. The machine was set at a 40 sec maximum burst interval dispersing 36 ml of chemical with an average droplet volume of 50%. Female adult Ae. albopictus were placed into cylindrical paper cages and adhered to poles at 5, 8, 10, 15, and 25 m from the center point of the machine. Control cages consisted of 1 cage placed at 5, 10, and 25 m. Control and treatment groups were left in the experiment area for 15 min. Initial knockdown after 15 min and 24 h mortality were documented. At 15 min post-treatment, Ae. albopictus displayed less than 50% knockdown. After 24 h, all treatment cages displayed greater than 90% mortality. Further bottle bioassays were conducted to determine the lowest chemical dose possible to achieve a lethal dose of 90%. A 1% dilution (10 ml Aqualuer 20-20 to 1,000 ml of polyether) of Aqualuer showed high mortality in the laboratory. However, after running 3 repetitions of a 1% dilution, there was no significant difference between the mortality of the mosquitoes at any of the distances 24 h post-treatment. This study indicates that the test machine would be an applicable and suitable machine for control of Ae. albopictus in enclosed spaces.