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.

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.

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.

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.

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.

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 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.

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.

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.

Synthesis and bioassay of improved mosquito repellents predicted from chemical structure.

Mosquito repellency data on acylpiperidines derived from the U.S. Department of Agriculture archives were modeled by using molecular descriptors calculated by CODESSA PRO software. An artificial neural network model was developed for the correlation of these archival results and used to predict the repellent activity of novel compounds of similar structures. A series of 34 promising N-acylpiperidine mosquito repellent candidates (4a-4q') were synthesized by reactions of acylbenzotriazoles 2a-2p with piperidines 3a-3f. Compounds (4a-4q') were screened as topically applied mosquito repellents by measuring the duration of repellency after application to cloth patches worn on the arms of human volunteers. Some compounds that were evaluated repelled mosquitoes as much as three times longer than N,N-diethyl-m-toluamide (DEET), the most widely used repellent throughout the world. The newly measured durations of repellency were used to obtain a superior correlation equation relating mosquito repellency to molecular structure.

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.

Expression of AeaHsp26 and AeaHsp83 in Aedes aegypti (Diptera: Culicidae) larvae and pupae in response to heat shock stress.

Immature mosquito development and survival of adults are highly sensitive to environmental temperature, which can alter gene expression during the mosquito life-cycle. To further understand how heat shock proteins are developmentally expressed in mosquitoes, we subjected first instar larvae, 16-h old pupae and female of Aedes aegypti (L.) (Diptera: Culicidae) to heat shock treatment for 0, 15, 30, 60, and 180 min at 23 and 42 degrees C. The heat shock protein genes AeaHsp26, AeaHsp83, and AeaHsc70 were examined by comparing relative transcript expression levels at 42 degrees C compared with 23 degrees C. Upregulated transcripts from heat shock treatment at 42 degrees C and control were further confirmed and quantified by quantitative real-time polymerase chain reaction. Data revealed that first instar larvae were more sensitive to heat shock treatment than pupae and adults (i.e., relative AeaHsp26 expression levels in larvae were 10-fold greater than in the females. AeaHsp83 expression levels in larvae, pupae and adults were upregulated 2- to 50-fold greater by heat shock treatment at 42 degrees C compared with 23 degrees C. AeaHsc70 expression levels in larvae, pupae and adults, however, were upregulated less than AeaHsp26 and AeaHsp83 at the higher temperature. Statistical analysis indicated that AeaHsp26 and AeaHsp83 genes were significantly upregulated in Ae. aegypti larvae and pupae after 15, 30, 60, and 180 min exposure to high temperature (42 degrees C). The current study has shown that AeaHsp26 and AeaHsp83 are important markers of stress and may function as critical proteins to protect and enhance survival of Ae. aegypti larvae and pupae.

Prallethrin-induced excitation increases contact between sprayed ultralow volume droplets and flying mosquitoes (Diptera: Culicidae) in a wind tunnel.

Female Culex quinquefasciatus Say (Diptera: Culicidae) mosquitoes were exposed to sublethal amounts of prallethrin, sumithrin, and piperonyl butoxide applied as ultralow volume (ULV) droplets in a wind tunnel. Mosquitoes were video recorded before, during, and after treatment, and the number and size of droplets on their bodies were later determined using a compound microscope. A positive correlation was found between mosquitoes that spent more time flying during the time of spraying and number of droplets on their bodies. Excitation, in the form of increased speed and duration of flight, was immediate in mosquitoes exposed to prallethrin, whereas exposure to sumithrin did not increase their exposure to the ULV droplets. The location of droplets on mosquitoes, the effects of droplet volume, and subsequent mortality are discussed.

Structure-activity relationships of 33 carboxamides as toxicants against female Aedes aegypti (Diptera: Culicidae).

Aedes aegypti L. is the primary vector of dengue and yellow fever viruses, and use of aerosolized insecticides is one of the primary ways to control this medically important mosquito. However, few new insecticides have been developed for mosquito control in recent years. As a part of our effort to search for new insecticides to control mosquitoes, toxicities of 33 carboxamides were evaluated against female A. aegypti by topical application. This group included nine different categories of compounds, namely benzamides, phenyl-propenamides, propanamides, butanamides, butenamides, pentanamides, pentenamides, hexanamides, and hexenamides, that exhibited varying levels of toxicity against this mosquito species. The most toxic compound tested was hexahydro-1-(1-oxohexyl)-1H-azepine, with a 24-h LD50 value of 0.4 microg per mosquito, whereas the most toxic compound at the LD95 level was N-ethyl-2-methyl-N-phenyl-benzamide (1.82 microg per mosquito). The least toxic compound was N,N-bis (2-methylpropyl)-3-phenyl-2-propenamide, with LD50 and LD95 values of 15.66 and 72.07 microg per mosquito, respectively. Results from this initial study may prove useful in guiding further carboxamide modifications for the development of potential new insecticides.

The biological activity of alpha-mangostin, a larvicidal botanic mosquito sterol carrier protein-2 inhibitor.

alpha-Mangostin derived from mangosteen was identified as a mosquito sterol carrier protein-2 inhibitor via high throughput insecticide screening, alpha-Mangostin was tested for its larvicidal activity against third instar larvae of six mosquito species, and the median lethal concentration values range from 0.84 to 2.90 ppm. The residual larvicidal activity of alpha-mangostin was examined under semifield conditions. The results indicated that alpha-mangostin was photolytic with a half-life of 53 min in water under full sunlight exposure. The effect of alpha-mangostin on activities of major detoxification enzymes such as P450, glutathione S-transferase, and esterase was investigated. The results showed that alpha-mangostin significantly elevated activities of P450 and glutathione S-transferase in larvae, whereas it suppressed esterase activity. Toxicity of alpha-mangostin against young rats was studied, and there was no detectable adverse effect at dosages as high as 80 mg/kg. This is the first multifaceted study of the biological activity of alpha-mangostin in mosquitoes. The results suggest that alpha-mangostin may be a lead compound for the development of a new organically based mosquito larvicide.

Identification and expression profile of multiple genes in response to magnesium exposure in Culex quinquefasciatus larvae.

Magnesium is crucial for baculovirus transmission in Culex nigripalpus (Theobald) and Culex quinquefasciatus (Say) larvae. However, the mechanistic role of magnesium in baculovirus transmission is unknown. To investigate the possible role of host response factors in baculovirus transmission, suppression subtractive hybridization was used to identify genes differentially transcribed after magnesium exposure in Cx. quinquefasciatus larvae. Suppression subtractive hybridization was performed in both directions enriching for cDNAs differentially transcribed between a nonmagnesium larval control and magnesium (15 mM MgSO4) treatment of Cx. quinquefasciatus larvae held for 1 h at 27 degrees C. Clones from differentially transcribed genes were evaluated by sequencing, and relative gene transcription levels were analyzed using quantitative real-time polymerase chain reaction quantitative real-time polymerase chain reaction. Target transcripts up/downregulated by magnesium included Cx. quinquefasciatus troponin C, isocitrate dehydrogenase, allergen, cytochrome b5, chymotrypsinogen, apolipophorins, tryptase gamma, carboxylesterase, prolylcarboxypeptidase, imaginal disc growth factor, aldehyde dehydrogenase, tropomyosin-1, chitotriosidase, heat shock protein 70 B2, inorganic phosphate cotransporter, and many other hypothetical protein genes. Magnesium can alter gene transcription in a vector mosquito population, and understanding this process can provide insight into the mechanistic role of magnesium in baculovirus transmission.

Novel carboxamides as potential mosquito repellents.

A model was developed using 167 carboxamide derivatives, from the United States Department of Agriculture archival database, that were tested as arthropod repellents over the past 60 yr. An artificial neural network employing CODESSA PRO descriptors was used to construct a quantitative structure-activity relationship model for prediction of novel mosquito repellents. By correlating the structure of these carboxamides with complete protection time, a measure of repellency based on duration, 34 carboxamides were predicted as candidate mosquito repellents. There were four additional compounds selected on the basis of their structural similarity to those predicted. The compounds were synthesized either by reaction of 1-acylbenzotriazoles with secondary amines or by reaction of acid chlorides with secondary amines in the presence of sodium hydride. The biological efficacy was assessed by duration of repellency on cloth at two dosages (25 and 2.5 micromol/cm2) and by the minimum effective dosage to prevent Aedes aegypti (L.) (Diptera: Culicidae) bites. One compound, (E)-N-cyclohexyl-N-ethyl-2-hexenamide, was superior to N,N-diethyl-3-methylbenzamide (deet) at both the high dosage (22 d versus 7 d for deet) and low dosage (5 d versus 2.5 d for deet). Only one of the carboxamides, hexahydro-1-(l-oxohexyl)-1H-azepine, had a minimum effective dosage that was equivalent or slightly better than that of deet (0.033 micromol/cm2 versus 0.047 micromol/cm2).

Mosquito vector biology and control in Latin America--a 20th symposium.

The 20th Annual Latin American Symposium presented by the American Mosquito Control Association (AMCA) was held as part of the 76th Annual Meeting in Lexington, KY, in March 2010. The principal objective, as for the previous 19 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 40 presentations that were given orally in Spanish or presented as posters by participants from 5 countries in Latin America, the United Kingdom, and the USA. Topics addressed in the symposium included: surveillance, chemical and biological control, and insecticide resistance associated with Aedes aegypti; distribution, behavior, and control of Culex; bionomics, ecology, and chemical and biological control of Anopheles vectors of malaria; insecticide resistance; and studies of dengue, West Nile virus, and Triatoma.