The Use of Microdispensers with Spatial Repellents for Personal Protection Against Mosquito Biting.

Mosquito-borne pathogens affect millions of people worldwide. This work describes a new method to deliver spatial repellents. Functional microdispensers (FMDs) were designed to deliver spatial repellents against mosquitoes. In vivo trials showed that FMDs protect human subjects against mosquitoes by reducing 70­90% of bites received, with a protection that lasted up to 4 weeks. FMDs can be cost-effectively implemented as wearable or field-dispensed devices for local area protection, defined as a confined geographical region.

Functional Micro-Dispensers based on Micro-Electro-Mechanical-Systems (MEMS) integrated with fabrics as functional materials to protect humans from mosquito feeding.

Functional Micro-Dispensers (FMDs) based on Micro-Electro-Mechanical-Systems (MEMS) were designed to deliver spatial repellents that reduce the ability of mosquitoes to feed on humans. FMDs were integrated with fabrics as functional materials for protection against mosquito bites. The use of MEMS devices provides an unprecedented control over the release kinetics by means of integration with electronics for selective and timely activation of each device to perform controlled release of pesticides in air. In addition, because MEMS manufacturing techniques evolved from the microelectronic industry, FMDs can be mass produced at very low cost. Trials using FMDs that contained transfluthrin improved protection against mosquito feeding in human subjects above that of permethrin-treated uniform fabric worn on the arm of the volunteer. The overall reduction in feeding was approximately 90% compared to the untreated fabric control, and about 50% reduction compared to the permethrin-treated fabric control. The devices were efficacious over course of 32 days. FMDs have the potential for a simple and cost-effective implementation for mass adoption as wearable devices integrated in fabrics as active functional materials.

Economic evaluation of an area-wide integrated pest management program to control the Asian tiger mosquito in New Jersey.

Aedes albopictus is the most invasive mosquito in the world, an important disease vector, and a biting nuisance that limits outdoor activities. Area-wide integrated pest management (AW-IPM) is the recommended control strategy. We conducted an economic evaluation of the AW-IPM project in Mercer and Monmouth Counties, New Jersey with a controlled design (AW-IPM vs. control) from 2009 through 2011. The study analyzed financial documents and staff time for AW-IPM and surveyed an average of 415 randomly chosen households in AW-IPM and control areas each fall from 2008 through 2011. Hours lost from yard and porch activities were calculated as differences between actual and potential hours of these activities in an average summer week if there had been no mosquito concerns. Net estimated benefits of AW-IPM were based on cross-over and difference-in-difference analyses. Reductions in hours lost were valued based on respondents' willingness to pay for a hypothetical extra hour free of mosquitoes spent on yard or porch activities and literature on valuation of a quality adjusted life year (QALY). The incremental cost of AW-IPM per adult was $41.18 per year. Number of hours lost due to mosquitoes in AW-IPM areas between the base year (2008) and the intervention years (2009-2011) declined by 3.30 hours per summer week in AW-IPM areas compared to control areas. Survey respondents valued this improvement at $27.37 per adult per summer week. Over the 13-week summer, an average adult resident gained 42.96 hours of yard and porch time, worth $355.82. The net benefit over the summer was $314.63. With an average of 0.0027 QALYs gained per adult per year, AW-IPM was cost effective at $15,300 per QALY gained. The benefit-cost ratio from hours gained was 8.64, indicating that each $1 spent on AW-IPM gave adults additional porch and yard time worth over $8.

Insecticide resistance status of United States populations of Aedes albopictus and mechanisms involved.

Aedes albopictus (Skuse) is an invasive mosquito that has become an important vector of chikungunya and dengue viruses. Immature Ae. albopictus thrive in backyard household containers that require treatment with larvicides and when adult populations reach pest levels or disease transmission is ongoing, adulticiding is often required. To assess the feasibility of control of USA populations, we tested the susceptibility of Ae. albopictus to chemicals representing the main insecticide classes with different modes of action: organochlorines, organophosphates, carbamates, pyrethroids, insect growth regulators (IGR), naturalytes, and biolarvicides. We characterized a susceptible reference strain of Ae. albopictus, ATM95, and tested the susceptibility of eight USA populations to five adulticides and six larvicides. We found that USA populations are broadly susceptible to currently available larvicides and adulticides. Unexpectedly, however, we found significant resistance to dichlorodiphenyltrichloroethane (DDT) in two Florida populations and in a New Jersey population. We also found resistance to malathion, an organophosphate, in Florida and New Jersey and reduced susceptibility to the IGRs pyriproxyfen and methoprene. All populations tested were fully susceptible to pyrethroids. Biochemical assays revealed a significant up-regulation of GSTs in DDT-resistant populations in both larval and adult stages. Also, ß-esterases were up-regulated in the populations with suspected resistance to malathion. Of note, we identified a previously unknown amino acid polymorphism (Phe → Leu) in domain III of the VGSC, in a location known to be associated with pyrethroid resistance in another container-inhabiting mosquito, Aedes aegypti L. The observed DDT resistance in populations from Florida may indicate multiple introductions of this species into the USA, possibly from tropical populations. In addition, the mechanisms underlying DDT resistance often result in pyrethroid resistance, which would undermine a remaining tool for the control of Ae. albopictus. Continued monitoring of the insecticide resistance status of this species is imperative.

Transcriptional analysis of four family 4 P450s in a Puerto Rico strain of Aedes aegypti (Diptera: Culicidae) compared with an Orlando strain and their possible functional roles in permethrin resistance.

A field strain of Aedes aegypti (L.) was collected from Puerto Rico in October 2008. Based on LD50 values by topical application, the Puerto Rico strain was 73-fold resistant to permethrin compared with a susceptible Orlando strain. In the presence of piperonyl butoxide, the resistance of Puerto Rico strain of Ae. aegypti was reduced to 15-fold, suggesting that cytochrome P450-mediated detoxification is involved in the resistance of the Puerto Rico strain to permethrin. To determine the cytochrome P450s that might play a role in the resistance to permethrin, the transcriptional levels of 164 cytochrome P450 genes in the Puerto Rico strain were compared with that in the Orlando strain. Of the 164 cytochrome P450s, 33 were significantly (P < 0.05) up-regulated, including cytochrome P450s in families four, six, and nine. Multiple studies have investigated the functionality of family six and nine cytochrome P450s, therefore, we focused on the up-regulated family 4 cytochrome P450s. To determine whether up-regulation of the four cytochrome P450s had any functional role in permethrin resistance, transgenic Drosophila melanogaster Meigen lines overexpressing the four family 4 P450 genes were generated, and their ability to survive exposure to permethrin was evaluated. When exposed to 5 microg per vial permethrin, transgenic D. melanogaster expressing CYP4D24, CYP4H29, CYP4J15v1, and CYP4H33 had a survival rate of 60.0 +/- 6.7, 29.0 +/- 4.4, 64.4 +/- 9.7, and 11.0 +/- 4.4%, respectively. However, none of the control flies survived the permethrin exposure at the same concentration. Similarly, none of the transgenic D. melanogaster expressing CYP4J15v1 or CYP4H33 ?5 survived when they were exposed to permethrin at 10 microg per vial. However, transgenic D. melanogaster expressing CYP4D24 and CYP4H29 had a survival rate of 37.8 +/- 4.4 and 2.2 +/- 2.2%, respectively. Taken together, our results suggest that CYP4D24 might play an important role in cytochrome P450-mediated resistance to permethrin.

Field evaluation of four spatial repellent devices against Arkansas rice-land mosquitoes.

Four commercially available spatial repellent devices were tested in a rice-land habitat near Stuttgart, AR, after semi-field level assessments had been made at the Center for Medical, Agricultural, and Veterinary Entomology, Agricultural Research Service, US Department of Agriculture in Gainesville, FL. OFF! Clip-On(metofluthrin), Mosquito Cognito (linalool), No-Pest Strip (dichlorvos), and ThermaCELL (d-cisltrans allethrin) were selected for this study from >20 candidate products. The units based on metofluthrin, linalool, or d-cisltrans allethrin significantly reduced captures of 1 or more of the mosquito species at surrogate human sites (unlit Centers for Disease Control and Prevention traps with CO2 and octenol). Among the mosquito species analyzed statistically (Anopheles quadrimaculatus, Culex erraticus, and Psorophora columbiae), there were significantly different responses (up to 84% reduction) to individual products, suggesting that combinations of certain spatial repellents might provide significantly greater protection.

Quantifying the impact of mosquitoes on quality of life and enjoyment of yard and porch activities in New Jersey.

The recent expansion of Aedes albopictus, a day-biting mosquito, to densely inhabited areas in the northeastern Atlantic states of the USA has dramatically increased the problem that mosquitoes create for urban and suburban residents. We quantified the impact of mosquitoes on residents' quality of life within the context of a comprehensive area-wide integrated pest management program to control Ae. albopictus in two counties (Mercer and Monmouth) in New Jersey. We interviewed residents of 121 randomly selected households in both counties between October and November 2010. We asked residents about their experience with mosquitoes in their neighborhood and the importance of the ability to relax outdoors without mosquitoes compared to other neighborhood characteristics (1 = not important, 5 = extremely important). We rated residents' utility based on paired comparisons to known states from the EuroQol health description system. The majority (54.6%) of respondents considered mosquitoes to be a problem. Respondents reported an average of 7.1 mosquito bites in a typical week during that summer. Mosquitoes prevented 59.5% of residents from enjoying their outdoor activities at least to some extent. Residents rated the mosquito acceptability (mean ± standard deviation) during that summer on a scale of 0 (mosquito invasion) to 100 (no mosquitoes) at 56.7±28.7, and their overall utility at 0.87±0.03. This is comparable to living with up to two risk factors for diabetes (i.e., abdominal obesity, body mass index of 28 or more, reported cholesterol problems, diagnosis of hypertension, or history of cardiovascular disease) or women experiencing menstrual disorders. Respondents rated the importance of enjoying outdoor activities without mosquitoes (4.69±0.80) comparable to that of neighborhood safety (4.74±0.80) and higher than that of a clean neighborhood (4.59±0.94). In conclusion, New Jersey residents reported that mosquitoes decreased their utility by 0.13, comparable to the loss from worrisome health risk factors, underscoring the importance of controlling this problem.

Mosquito vector biology and control in latin america-a 24th symposium.

The 24th Annual Latin American Symposium presented by the American Mosquito Control Association (AMCA) was held as part of the 80th Annual Meeting in Seattle, WA, in February 2014. The principal objective, for the previous 23 symposia, was to promote participation in the AMCA by vector control specialists, public health workers, and academicians from Latin America. This publication includes summaries of 26 presentations that were given orally in Spanish or presented as posters by participants from Colombia, Mexico, and the USA. Topics addressed in the symposium included: surveillance, ecology, chemical control, studies of dengue viruses, and insecticide resistance associated with Aedes aegypti; Anopheles vectors of malaria; essential oils; and ethnic groups and vector-borne diseases.

Mosquito vector biology and control in Latin America--a 23rd symposium.

The 23rd Annual Latin American Symposium presented by the American Mosquito Control Association (AMCA) was held as part of the 79th Annual Meeting in Atlantic City, NJ, in February 2013. The principal objective, as for the previous 22 symposia, was to promote participation in the AMCA by vector control specialists, public health workers, and academicians from Latin America. This publication includes summaries of 49 presentations that were given orally in Spanish or presented as posters by participants from Colombia, Mexico, and the USA. Topics addressed in the symposium included: surveillance, ecology, chemical and biological control, and insecticide resistance associated with Aedes aegypti; surveillance and control of Anopheles vectors of malaria; and studies of dengue and West Nile viruses, Chagas disease, and Lutzomyia.

Behavioral responses of two dengue virus vectors, Aedes aegypti and Aedes albopictus (Diptera: Culicidae), to DUET and its components.

Ultralow volume droplets of DUET, prallethrin, and sumithrin at a sublethal dose were applied to unfed (nonbloodfed) and bloodfed female Aedes aegypti L. and Aedes albopictus (Skuse) in a wind tunnel. Control spray droplets only contained inert ingredients. Individual mosquitoes were videotaped before, during, and after spraying and various behaviors analyzed. During the spray periods of all three pesticide treatments, mosquitoes spent a greater percentage of time moving, and the distance moved was greater than for mosquitoes in the control treatments. In the postspray period, the percent of time moving increased for mosquitoes exposed to all pesticide treatments compared with the controls. After treatment, all females spent more time walking compared with controls, with unfed Ae. aegypti females walking more after exposure to DUET and sumithrin than after exposure to prallethrin and the control. Pesticide exposure increased flying in both species. Sumithrin exposure increased activity and velocity of unfed mosquitoes more than bloodfed mosquitoes. DUET and sumithrin treatments enhanced activity of Ae. aegypti females more than Ae. albopictus females.

Promising Aedes aegypti repellent chemotypes identified through integrated QSAR, virtual screening, synthesis, and bioassay.

Molecular field topology analysis, scaffold hopping, and molecular docking were used as complementary computational tools for the design of repellents for Aedes aegypti, the insect vector for yellow fever, chikungunya, and dengue fever. A large number of analogues were evaluated by virtual screening with Glide molecular docking software. This produced several dozen hits that were either synthesized or procured from commercial sources. Analysis of these compounds by a repellent bioassay resulted in a few highly active chemicals (in terms of minimum effective dosage) as viable candidates for further hit-to-lead and lead optimization effort.

Area-wide management of Aedes albopictus. Part 2: gauging the efficacy of traditional integrated pest control measures against urban container mosquitoes.

BACKGROUND: Aedes (Stegomyia) albopictus (Skuse) is an important disease vector and biting nuisance. During the 2009 active season, six ∼1000-parcel sites were studied, three in urban and three in suburban areas of New Jersey, United States, to examine the efficacy of standard integrated urban mosquito control strategies applied area wide. Active source reduction, larviciding, adulticiding and public education (source reduction through education) were implemented in one site in each county, an education-only approach was developed in a second site and a third site was used as an untreated experimental control. Populations were surveyed weekly with BG-Sentinel traps and ovitraps. RESULTS: A substantial reduction in Ae. albopictus populations was achieved in urban sites, but only modest reductions in suburban sites. Education alone achieved significant reductions in urban adult Ae. albopictus. Egg catches echoed adult catches only in suburban sites. CONCLUSIONS: There are significant socioeconomic and climatic differences between urban and suburban sites that impact upon Ae. albopictus populations and the efficacy of the control methods tested. An integrated pest management approach can affect abundances, but labor-intensive, costly source reduction was not enough to maintain Ae. albopictus counts below a nuisance threshold. Nighttime adult population suppression using truck-mounted adulticides can be effective. Area-wide cost-effective strategies are necessary.

Insecticidal, repellent and fungicidal properties of novel trifluoromethylphenyl amides.

Twenty trifluoromethylphenyl amides were synthesized and evaluated as fungicides and as mosquito toxicants and repellents. Against Aedes aegypti larvae, N-(2,6-dichloro-4-(trifluoromethyl)phenyl)-3,5-dinitrobenzamide (1e) was the most toxic compound (24 h LC50 1940 nM), while against adults N-(2,6-dichloro-4-(trifluoromethyl)phenyl)-2,2,2-trifluoroacetamide (1c) was most active (24 h LD50 19.182 nM, 0.5 µL/insect). However, the 24 h LC50 and LD50 values of fipronil against Ae. aegypti larvae and adults were significantly lower: 13.55 nM and 0.787 × 10(-4) nM, respectively. Compound 1c was also active against Drosophila melanogaster adults with 24 h LC50 values of 5.6 and 4.9 µg/cm(2) for the Oregon-R and 1675 strains, respectively. Fipronil had LC50 values of 0.004 and 0.017 µg/cm(2) against the two strains of D. melanogaster, respectively. In repellency bioassays against female Ae. aegypti, 2,2,2-trifluoro-N-(2-(trifluoromethyl)phenyl)acetamide (4c) had the highest repellent potency with a minimum effective dosage (MED) of 0.039 µmol/cm(2) compared to DEET (MED of 0.091 µmol/cm(2)). Compound N-(2-(trifluoromethyl)phenyl)hexanamide (4a) had an MED of 0.091 µmol/cm(2) which was comparable to DEET. Compound 4c was the most potent fungicide against Phomopsis obscurans. Several trends were discerned between the structural configuration of these molecules and the effect of structural changes on toxicity and repellency. Para- or meta- trifluoromethylphenyl amides with an aromatic ring attached to the carbonyl carbon showed higher toxicity against Ae. aegypti larvae, than ortho- trifluoromethylphenyl amides. Ortho- trifluoromethylphenyl amides with trifluoromethyl or alkyl group attached to the carbonyl carbon produced higher repellent activity against female Ae. aegypti and Anopheles albimanus than meta- or para- trifluoromethylphenyl amides. The presence of 2,6-dichloro- substitution on the phenyl ring of the amide had an influence on larvicidal and repellent activity of para- trifluoromethylphenyl amides.

Mosquito vector biology and control in Latin America--a 22nd symposium.

The 22nd Annual Latin American Symposium presented by the American Mosquito Control Association (AMCA) was held as part of the 78th Annual Meeting in Austin, TX, in February 2012. The principal objective, as for the previous 21 symposia, was to promote participation in the AMCA by vector control specialists, public health workers, and academicians from Latin America. This publication includes summaries of 21 presentations that were given orally in Spanish or presented as posters by participants from Mexico, Colombia, Venezuela, and the USA. Topics addressed in the symposium included surveillance, chemical control, insecticide resistance, and genetics associated with Aedes aegypti; food sources and control of Culex; taxonomy, surveillance, and control of Anopheles vectors of malaria; and studies of dengue virus and Leishmania.

Willingness-to-pay for an area-wide integrated pest management program to control the Asian tiger mosquito in New Jersey.

Using contingent valuation we estimated the perceived value of an area-wide integrated pest management program for the Asian tiger mosquito, Aedes albopictus, implemented in Monmouth and Mercer counties, NJ. We estimated residents' maximum willingness-to-pay and perceived monetary benefits (willingness-to-pay excluding residents who protested all types of payments) and payment modality through a telephone survey of 51 randomly selected households. The mean (+/- SE) perceived monetary benefits for an enhanced mosquito abatement program was $9.54 +/- 2.90 per capita per year. Most respondents would have been willing to pay through taxes (35%) or charitable donations (6%) starting then, or through one of these approaches in the future (43%), whereas 16% were completely unwilling to pay any additional costs whatsoever. We projected that the perceived monetary benefits to the counties' 1.01 million residents for an enhanced mosquito control program would be $9.61 million annually. Thus, collectively residents perceived monetary benefits of 3.67 times the combined 2008 annual operating costs of the counties' existing mosquito control programs of $2.61 million.

Countering a bioterrorist introduction of pathogen-infected mosquitoes through mosquito control.

The release of infected mosquitoes or other arthropods by bioterrorists, i.e., arboterrorism, to cause disease and terror is a threat to the USA. A workshop to assess mosquito control response capabilities to mount rapid and effective responses to eliminate an arboterrorism attack provided recommendations to improve capabilities in the USA. It is essential that mosquito control professionals receive training in possible responses, and it is recommended that a Council for Emergency Mosquito Control be established in each state to coordinate training, state resources, and actions for use throughout the state.

Source reduction behavior as an independent measurement of the impact of a public health education campaign in an integrated vector management program for the Asian tiger mosquito.

The goal of this study was to evaluate the effectiveness of a public health educational campaign to reduce backyard mosquito-larval habitats. Three communities each, within two New Jersey counties, were randomly selected to receive: (1) both education and mosquito control, (2) education only, and (3) no education or mosquito control. Four separate educational events included a 5-day elementary school curriculum in the spring, and three door to door distributions of educational brochures. Before and after each educational event, the numbers of mosquito-larval container habitats were counted in 50 randomly selected homes per study area. Container surveys allowed us to measure source reduction behavior. Although we saw reductions in container habitats in sites receiving education, they were not significantly different from the control. Our results suggest that traditional passive means of public education, which were often considered the gold standard for mosquito control programs, are not sufficient to motivate residents to reduce backyard mosquito-larval habitats.

Changes in host-seeking behavior of Puerto Rican Aedes aegypti after colonization.

The effects of colonization on host-seeking behavior of mosquitoes was examined by comparing attraction responses of newly colonized Aedes aegypti (L.) from field-collected eggs in Puerto Rico to that of the Gainesville (Florida) strain, originally from Orlando (Florida) and in colony since 1952. Females from the Orlando and the F0 through F10 generations of the Puerto Rico strain were evaluated using attractant odors in a triple-cage dual-port olfactometer. Two attractant sources were used: odors from the hand of a volunteer and a standard blend of L-lactic acid, acetone, and dimethyl disulfide. Convergence of the percentage of attraction responses occurred around the F4-F6 generations of the Puerto Rico strain. Both the Orlando and Puerto Rico strains exhibited similar responses for tests with the remaining F7-F10 generations. A temporal effect on mosquito responses was observed for both strains regardless of the attractant blend used in tests. This study indicates that Ae. aegypti host-seeking behavior changes significantly over the first four to six generations after introduction into the laboratory, whereas the field-collected strain increases in attraction response until it stabilizes at a new level.

Identification and transcription profiling of trypsin in Aedes taeniorhynchus (Diptera: Culicidae): developmental regulation, blood feeding, and permethrin exposure.

The cDNA of a trypsin gene from Aedes (Ochlerotatus) taeniorhynchus (Weidemann) was cloned and sequenced. The full-length mRNA sequence (890 bp) for trypsin from Ae. taeniorhynchus (AetTryp1) was obtained, which encodes an open reading frame of 765 bp (i.e., 255 amino acids). To detect whether AetTryp is developmentally regulated, a quantitative real-time polymerase chain reaction was used to examine AetTrypl mRNA expression levels in different developmental stages of Ae. taeniorhynchus. AetTryp1 was expressed at low levels in egg, larval, and pupal stages, but was differentially expressed in adult Ae. taeniorhynchus, with highest levels found in 5-d-old female adults when compared with teneral adults. In addition, AetTryp1 mRNA expression differed between sexes, with expression levels much lower in males. However, in both males and females, there was a significant increase in AetTryp1 transcription levels as age increased and peaked in 5-d-old adults. AetTrypl expressed in 5-d-old female Ae. taeniorhynchus significantly increased after 30 min postblood feeding compared with the control. The AetTryp1 mRNA expression in 5-d-old female Ae. taeniorhynchus was affected by different concentrations of permethrin.

Area-wide management of Aedes albopictus: choice of study sites based on geospatial characteristics, socioeconomic factors and mosquito populations.

BACKGROUND: Aedes (Stegomyia) albopictus (Skuse), the Asian tiger mosquito, is an introduced invasive species in the United States that is responsible for a significant proportion of service requests to local mosquito control programs. This container-utilizing mosquito is refractory to standard mosquito abatement measures in the United States. This study is part of a USDA-ARS project to develop an area-wide management strategy for Ae. albopictus. The goal was to identify three study sites, similar in socioeconomic parameters, geography and Ae. albopictus abundance, in urban and suburban areas in Mercer and Monmouth counties in New Jersey. Prior service requests and light trap counts and also detailed county maps were used to chose nine preliminary sites (four in Mercer and five in Monmouth) where weekly surveillance for Ae. albopictus was performed throughout the 2008 active season. RESULTS: Although outliers were detected, socioeconomic variables in the study sites within each county were fairly consistent. Ae. albopictus abundance was associated with poverty levels and had the highest maxima in Mercer, although average mosquito abundance was similar in urban Mercer and suburban Monmouth. CONCLUSION: Three study sites in each county were identified for future studies. The summer-long surveillance also revealed socioeconomic variables critical for the development of integrated mosquito management.