A total of 28 species of mosquitoes was collected in dry-ice–baited light traps on Key Largo, Florida during 2002–2004. Total number of mosquitoes increased each year during the study.

Parts 1 to 6 of this series on the mosquitoes of the mangrove forests of India dealt with the mosquito species recorded in the mangroves of Bhitarkanika, Sundarbans, Andaman and Nicobar islands, Coringa, Chorao and Vikhroli, and Kundapur and Kannur. This concluding part provides an overview of the distribution of the mosquito species in different mangrove forests, including the mangroves of Muthupet in Tamilnadu and the mangroves of Gulf of Kutch and Gulf of Kambhat in Gujarat, species collected as larvae, species in relation to the salinity of the larval habitats, species landing on humans for feeding in the mangroves, and the impact of habitat degradation on species diversity.

To verify the genetic proximity of Anopheles sinensis strains in Korea, China, Japan, and Thailand, we established 5 iso-female lines, and investigated their genetic relationships by analyzing crossing tests and comparing the DNA sequences of rDNA internal transcribed spacer 2 (ITS2) and mitochondrial cytochrome c oxidase subunit 2 (CO2). The results of reciprocal and F₁-hybrid crosses between Thailand Form B (X, Y2) with Japanese Form A (X, Y1), Japanese Form B with Korean Form B, and Korean Form B with Chinese Form B indicated that these strains were genetically compatible, providing viable progenies and completely synaptic polytene chromosomes. The sequences of ITS2 and CO2 among 10 Asian An. sinensis strains were almost identical to each other, and the sequence variation was very low (<0.6% in both regions). Based on the above results, it was confidently concluded that An. sinensis is a single cosmopolitan species even though its geographical distribution ranges are comparatively wide (>2,000 mi) and that the malaria-vector abilities of the various strains may vary depending on their geographical origins.

Monitoring reproductive rates in experiments involving aedine mosquitoes is tedious and time demanding. Here, we demonstrate a protocol for rapid estimation of aedine mosquito egg number. The protocol uses ImageJ, a publicly available image analysis program developed at the US National Institutes of Health. The method relies upon the oviposition behavior of Aedes (i.e., ovipositing on a moist substrate instead of water surface) and upon the contrast between dark-colored aedine eggs and a light, uniformly colored paper that is used as an oviposition substrate. The results for 3 Aedes species show that, following the generation of separate calibration curves for each species, the protocol allows for the accurate and repeatable estimation of samples containing hundreds of aedine eggs. We discuss the use of the protocol for monitoring immature aedine populations in both laboratory and field applications.

The rapid spread of West Nile viral activity across North America since its discovery in 1999 illustrates the potential for an exotic arbovirus to be introduced and widely established across North America. Rift Valley fever virus (RVFV) 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. However, little is known about the potential for North American mosquitoes to transmit this virus should it be introduced into North America. Therefore, we evaluated selected mosquito species from the southeastern United States for their ability to serve as potential vectors for RVFV. Mosquitoes were fed on adult hamsters inoculated 1 day previously with RVFV. These mosquitoes were tested for infection and ability to transmit RVFV after incubation at 26°C for 7–21 days. None of the species tested (Aedes taeniorhynchus, Ae. vexans, Culex erraticus, Cx. nigripalpus, Cx. quinquefasciatus, and Cx. salinarius) were efficient vectors after they fed on hamsters with viremias ranging from 10^4.1 to 10^6.9 plaque-forming units (PFU)/ml. However, Ae. taeniorhynchus, Ae. vexans, and Cx. erraticus all developed disseminated infections after they fed on hamsters with viremias between 10^8.5 and 10^10.2 PFU/ml, and both Ae. vexans and Cx. erraticus transmitted RVFV by bite. These studies illustrate the need to identify the ability of individual mosquito species to transmit RVFV so that appropriate decisions can be made concerning the application of control measures during an outbreak.

We report that an aqueous solution containing 10⁻³ M adenosine triphosphate (ATP) and citrate-phosphate-dextrose-adenine (CPDA-1) can effectively replace transfusable human red blood cells in an in vitro Klun and Debboun bioassay system for evaluating chemicals for mosquito feeding-deterrent activity, using either Aedes aegypti or Anopheles stephensi. These species fed with similar avidity through collagen membrane covering aqueous 10⁻³ M ATP plus CPDA-1 or red blood cells in CPDA-1 supplemented with ATP. In a 2nd experiment, we evaluated the feeding-deterrent activity of N,N-diethyl-3-methylbenzamide and a newly discovered natural product chemical, (-)-isolongifolenone, against these 2 mosquito species. We found that the feeding-deterrent efficacy of the 2 chemicals was similar whether the feeding stimulant was red blood cells supplemented with ATP or ATP alone with CPDA-1. Since the use of human red blood cells in bioassays raises important health and logistic issues, aqueous ATP with CPDA-1 is a reasonable alternative to human blood cells for routine in vitro chemical screening.

An investigation of the biological effect of catnip oil (Nepeta cataria L.) on the behavioral response of field collected Aedes aegypti and Anopheles harrisoni was conducted using an automated excito-repellency test system. Aedes aegypti showed significantly higher escape rates from the contact chamber at 5% catnip oil compared to other concentrations (P < 0.05). With Anopheles harrisoni, a high escape response was seen at 2.5% catnip oil from the contact chamber, while in the noncontact chamber a higher escape response was observed at a concentration of 5%. Results showed that this compound exhibits both irritant and repellent actions.

We conducted trials in Cairns, Australia, to examine if novel updraft light traps collected significantly more mosquitoes than the Centers for Disease Control and Prevention (CDC) model 512 miniature light trap. Two new updraft traps, the Northern Australia Quarantine Strategy (NAQS) Mozzie Trap and a CDC updraft trap, both collected significantly more mosquitoes than the standard CDC light trap, with a mean CDC Trap Index (trap collections relative to paired standard CDC light trap collections) of 3.3 and 2.3, respectively. These traps both had large horizontal suction areas that increased the probability that attracted mosquitoes entered the trap updraft. However, if the CO₂ source was located within the updraft of the CDC updraft trap, mosquito collections decreased considerably, indicating that placement of the bait is critical to trap performance. Creating an updraft by simply inverting the CDC trap body did not increase collections. The Mosquito Magnet X trap also did not collect significantly more mosquitoes than the CDC trap. Two CDC light traps sharing a 600 ml CO₂/min gas line collected ca. 50% more mosquitoes than a single CDC trap baited with 600 ml CO₂/min, suggesting that a single gas source could be used on a trap line consisting of multiple trap units. These studies suggest that the optimal trap design should incorporate a CO₂ release system that lures mosquitoes to a large updraft within a bowl-shaped trap intake.

The Mosquito Sentinel 360™ (MS) trap was evaluated in back yards of 6 residences in and near Gainesville, FL. Carbon dioxide and blue-light–emitting diode-modified Centers for Disease Control and Prevention (CDC) traps were utilized as a human substitute host to provide a measure of the effectiveness of the MS trap in reducing mosquito presence at the back door area of the houses. At 5 of the 6 residences, a MS trap was operated with or without carbon dioxide, Lurex3 octenol, or an experimental attractant. All MS traps were operated in tandem with a CDC trap, with the 6th site occupied only by a CDC trap. Over 67,000 mosquitoes from 19 species were recovered over the 12 trial days. Of these species, Anopheles crucians, An. quadrimaculatus, Coquillettidia perturbans, Culex erraticus, and Mansonia titillans were the predominant, nuisance, and medically important species captured and accounted for >92% of specimens recovered. Overall, attractant-modified MS traps captured more mosquitoes than the paired CDC trap. Carbon dioxide was found to greatly increase the capture of mosquitoes; however, the Lurex3 octenol combination was significant with Cq. perturbans. All attractant-modified MS traps captured more Cx. erraticus and Ma. titillans than did the CDC, CO₂-baited host mimic traps. The use of the MS 360 trap will undoubtedly require the addition of baits and perhaps the inclusion of a host masking or repellent approach to ensure protection from nuisance mosquitoes in residential environments.

Two separate but related studies were conducted regarding management of Anopheles quadrimaculatus larval populations in commercial rice fields near Cleveland, MS, in 2004. Study 1 was to evaluate the effectiveness of 2 treatments of aerially applied ultra-low volume applications of Bacillus thuringiensis var. israelensis (Bti) against An. quadrimaculatus larvae in dense, high-canopy mid- to late-season rice crop. Study 2 was to investigate the effect of preflood treatments of lambda-cyhalothrin (Karate®), which is commonly used against rice water weevil (Lissorhoptrus oryzophilus), on An. quadrimaculatus larvae. Excellent initial, but short residual control (>99% control 1 day after treatment) was observed in the Bti-treated fields in both mid- and late-season rice. Little or no effect on mosquito larvae was observed in the lambda-cyhalothrin–treated fields. Results indicate that Bti can be effectively used by mosquito management personnel to control larval populations of An. quadrimaculatus in late-season rice fields; however, lambda-cyhalothrin did not effectively control larval An. quadrimaculatus when applied preflood to rice fields.

The efficacy and persistence of 2 bacterial larvicides, Vectobac-DT (Bacillus thuringiensis israelensis [Bti]) and CulinexCombi (Bti and Bacillus sphaericus [Bs]), were tested against Anopheles gambiae and Culex quinquefasciatus in temporarily unused swimming pools with rainwater in Malindi, Kenya. Pre- and posttreatment larval densities were recorded by sampling with the standard WHO dipping technique for 8 consecutive days. The larvicides were applied to the pools with a knapsack sprayer. The data showed that Vectobac-DT was highly effective against early instars of An. gambiae with 89% reduction within 24 h but not as effective against the early stages of Cx. quinquefasciatus with reduction of only 46%. CulinexCombi resulted in high mortalities to early instars of both species with over 97% reduction within 24 h, but showed a drastic reduction 48 h after application. Both Vectobac-DT and CulinexCombi were highly effective against late instars of both species, whereby Vectobac-DT persisted much longer than CulinexCombi. Anopheles gambiae was found to be more susceptible to both larvicides than Cx. quinquefasciatus. By their high efficacy and good persistence against mosquito larvae, both Vectobac-DT and CulinexCombi can be recommended for use in integrated mosquito control programs.

Backpack sprayer applications of permethrin, d-phenothrin, and resmethrin to vegetation and plants at Anastasia Island, St. Augustine, FL, were evaluated for duration of residual efficacy against adult mosquitoes. All treatments produced 100% mortality (24 h) of mosquitoes in test cages placed within the vegetation. At 48 h and 1 wk posttreatment, insecticide treatments resulted in 70–100% reduction of adult mosquitoes caught by Centers for Disease Control and Prevention traps baited with 1-octen-3-ol. Insecticide residues in excised leaves from both treated and control areas of the study sites were evaluated against adult female Culex quinquefasciatus by laboratory bioassay. Permethrin produced 90% mortality up to 1 wk postapplication. Both d-phenothrin and resmethrin produced nearly this level of mortality for a much shorter duration of <48 h postapplication, with residual effects dropping significantly thereafter. Average insecticide concentrations in leaves were quantified by gas chromatography/mass spectroscopy, and some correlation was observed between chemical and biological results.

Droplet size spectra from different sprayers used to generate insecticide-laden fogs for controlling flying insects were measured by a laser diffraction instrument and Teflon-coated slides. The objectives of this work were to present not only information on spray-system droplet size generated by different sprayers, but to compare methodologies by which other similar systems can be evaluated and give applicators sprayer-system performance data. Data from 45 replicated spray tests, comprising 11 sprayers and 5 pesticides, showed a wide range in the droplet size spectra produced. The volume median diameter measurements ranged from 2.6 to 75.5 μm for diesel-diluted sprays and from 27.9 to 59.9 μm for water-diluted sprays. Similarly, the percent volume <20 μm ranged between 12.0–100% and 8.5–30.7%, for diesel- and water-diluted sprays, respectively. The droplet sizes measured by the swinging slide and laser diffraction methods were not consistent. The information presented aids users in sprayer selection and operation to produce the specific droplet size spectra required for a particular application.

Several manufacturers and pest management companies have begun to market and install outdoor automatically timed insecticide application systems that claim to provide an envelope of protection against host-seeking mosquitoes within a defined area, e.g., residential backyards. A typical system consists of a multi-gallon reservoir attached to a continuous loop of plastic tubing with multiple single spray head nozzles. Nozzles are usually placed along the perimeter of a backyard in landscaping or other areas suitable for mosquito harborage. This array is then connected to a programmable electric pump set to automatically apply an insecticide at predetermined intervals. An operational field study was conducted to evaluate this technology using previously installed MistAway© systems at 3 residences in northwestern Florida. This system applied a mist-like application of 0.05% AI synergized pyrethrins for 45 sec at dawn and again at dusk in each backyard. Twice-weekly collections from ABC suction light traps, baited with carbon dioxide, were used as the evaluation tool. Female mosquitoes from treatment backyards were compared with trap collections from 3 backyards without automatic misting systems used as controls. We found that weekly mosquito reduction was highly variable and ranged from 98% to 14% during the 35-wk study. Because the primary method of reduction by these application systems was not well understood, a MistAway system was installed in an outdoor simulated residential backyard to determine exposure pathway under controlled conditions with field cage and excised-leaf bioassays. Using laboratory-reared females of Aedes albopictus and Culex quinquefasciatus in those assays, we found that reduction by the MistAway system was primarily achieved by direct exposure of the mosquitoes to the insecticide application and not from residual deposits on treated vegetation.

Spring acaricide applications directed against nymphal Ixodes scapularis have been shown to be effective, but are perceived by the public as having significant adverse environmental impacts, particularly against nontarget organisms. Targeting the adult stage of I. scapularis in the fall would hypothetically result in indirect control of subsequent subadult stages while avoiding other arthropods that are typically inactive during this period. We demonstrate that single fall applications of deltamethrin for 3 consecutive years immediately reduced fall questing adults, while also rapidly reducing the abundance of all postembryonic stages. Deltamethrin applied to the shrub-layer vegetation resulted in levels of control between 97.1% and 100% at 7 days postapplication. Repeated applications against the reproductive stage of I. scapularis progressively reduced the abundance of larvae and nymphs in treated plots, reaching 91.4% and 100% by the conclusion of the study.

The 18th Annual Latin American symposium presented by the American Mosquito Control Association (AMCA) was held as part of the 74th Annual Meeting in Sparks, NV, in March 2008. The principal objective, as for the previous 17 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 35 presentations that were given orally in Spanish or presented as posters by participants from 5 countries in Latin America and Puerto Rico. Topics addressed in the symposium included surveillance, chemical and biological control, insecticide resistance, and repellency of Aedes aegypti; distribution, behavior, transmission of West Nile virus, and control of Culex; bionomics, ecology, and chemical and biological control of Anopheles vectors of malaria; insecticide resistance; and studies of Triatoma and Rhodnius.

Aedes japonicus japonicus was collected via gravid trapping in Wisconsin in the summers of 2004 and 2005 at Fort McCoy, Monroe County. Subsequently, in the summer of 2007, Ae. japonicus was captured in a human landing catch in Dane County, Madison, WI. Additional collections were made at this site in the spring of 2008. Invasion is in progress, but significant population increases have not yet been confirmed.

Culex coronator, a mosquito species common to the American tropics, has been recently documented from a number of temperate areas in the USA. Since 2002 specimens have been reported for the first time from Oklahoma, Louisiana, Mississippi, Alabama, and Florida. Here we provide new collection records for Cx. coronator in east-central Alabama. In October 2007, 2 larvae of Cx. coronator were collected from an artificial container in Tuskegee National Forest in Macon County, AL. The distribution of Cx. coronator in the USA seems to be expanding at a prodigious rate, for reasons that remain unclear.

In 2007, adult Culex coronator were collected in Chatham County, Georgia, and Jasper County, South Carolina, during nuisance and disease vector surveillance efforts. A total of 75 specimens of this species were collected at 8 widely separated locations in Chatham County, Georgia, and 4 closely situated sites in Jasper County, South Carolina. These represent the first Atlantic coastal records of this species in Georgia and the first confirmed records of Cx. coronator in South Carolina.

We describe methodology used for the laboratory colonization of Aedes japonicus japonicus, an exotic mosquito species native to eastern Asia and first collected in New Jersey as larvae in 1999. We created a free mating colony in 2000 that readily bloodfed on restrained bobwhite quail (Colinus virginianus). A larval diet of finely ground Purina Lab Diet© dissolved in dechlorinated water has proven acceptable. This is the first report of Ae. j. japonicus colonization from mosquitoes collected in the United States.

A novel device for the containment and precise release of aromatic substances is described. The device consists of a threaded-tubular polyvinyl chloride chamber (and screw-top cap) with ports for introduction and release of gaseous compounds. This chamber is inexpensive, easy to assemble, and useful for evaluating the combined release of carbon dioxide and aromatic hygroscopic substances as mosquito attractants in field studies.

Laboratory studies were conducted to evaluate the effects of treating military canvas tent fabric with bifenthrin and permethrin on the survival of the eggs and larvae of Aedes aegypti. Gravid female Ae aegypti were able to oviposit on tent canvas treated with either bifenthrin or permethrin. However, none of the eggs laid on treated canvas hatched, and no larvae added to water in treated trays survived. Low residual concentrations of bifenthrin and permethrin on treated canvas prevented the development of eggs and larvae of Ae. aegypti. Inhibition of bloodfeeding was shown when Ae. aegypti adults were exposed to lower concentrations (10–50% of operational concentrations) of bifenthrin- and permethrin-treated canvas tent fabric. These experiments have shown that military tent canvas treated with either bifenthrin or permethrin can reduce the development of Ae. aegypti eggs and larvae and reduce bloodfeeding success of adults.

In an attempt to find diurnal resting sites of adult phlebotomine sand flies, potential phlebotomine adult habitats were aspirated in the village of Bahrif in Aswan, Egypt. During this survey, sand flies were aspirated from low (30–45 cm high) irregular piles of mud bricks found under high date palm canopies between the village and the Nile River. There were 5 ♂♂ and 7 ♀♀ of Phlebotomus papatasi and 3 ♂♂ of Sergentomyia schwetzi. Six of the 7 aspirated females were engorged with blood. A total of 78 sand flies was captured on 3 glue boards placed overnight on the ground next to the mud bricks. Attempts to aspirate sand flies from adjacent walls and plants were unsuccessful. The identification of diurnal resting sites in less structured habitats may ultimately lead to more effective adult sand fly control.

For mosquito vector population monitoring, a new commercial trap, Mosquito Magnet™ Pro (MM-PRO), was tested for its usefulness in Goa, India. Anopheles stephensi was tested for the presence of Plasmodium sporozoite infection in the salivary glands. Using the MM-PRO 24 h a day for 34 days, 2,329 mosquitoes belonging to 16 species were collected. These included 6 species each of the genera Anopheles and Culex, 2 species of Aedes, and 1 each of Mansonia and Armigeres. Most (91%) of the mosquitoes caught were females. Among these the number and percentage of each species were Anopheles stephensi 59 (2.78%), Culex quinquefasciatus 1013 (47.78%), Culex vishnui 551 (26.0%), Mansonia uniformis 216 (10.19%), and Aedes albopictus 1 (0.04%). Of the 54 An. stephensi females tested for the presence of circumsporozoite protein (CSP) by an ELISA technique, 1 was found to be Plasmodium falciparum CSP positive. The MM-PRO device was found useful for mosquito population sampling in the urban setting of Goa.

Monitoring of resistance of Aedes aegypti to temephos was implemented in the provinces of Formosa and Misiones, Argentina, as a response to the need to improve the vigilance for the dengue vector in areas of high risk of dengue. Eggs collected in each locality were reared, and susceptibility to temephos was assayed using larval bioassays. A weak decrease in susceptibility of larvae to temephos was observed in Clorinda and Puerto Iguazú, indicating an incipient resistance with a resistance ratio of 3. No control failures have been observed yet, and this program should allow the early detection of a real problem in our country.