Isolation of viruses from mosquitoes (Diptera: Culicidae) collected in the Amazon Basin region of Peru.

As part of a comprehensive study on the ecology of arthropod-borne viruses in the Amazon Basin region of Peru, we assayed 539,694 mosquitoes captured in Loreto Department, Peru, for arboviruses. Mosquitoes were captured either by dry ice-baited miniature light traps or with aspirators while mosquitoes were landing on human collectors, identified to species, and later tested on Vero cells for virus. In total, 164 virus isolations were made and included members of the Alphavirus (eastern equine encephalomyelitis, Trocara, Una, Venezuelan equine encephalomyelitis, and western equine encephalomyelitis viruses), Flavivirus (Ilheus and St. Louis encephalitis), and Orthobunyavirus (Caraparu, Itaqui, Mirim, Murutucu, and Wyeomyia viruses) genera. In addition, several viruses distinct from the above-mentioned genera were identified to the serogroup level. Eastern equine encephalomyelitis virus was associated primarily with Culex pedroi Sirivanakarn & Belkin, whereas Venezuelan equine encephalomyelitis virus was associated primarily with Culex gnomatos Sallum, Huchings & Ferreira. Most isolations of Ilheus virus were made from Psorophora ferox (Von Humboldt). Although species of the Culex subgenus Melanoconion accounted for only 45% of the mosquitoes collected, 85% of the virus isolations were made from this subgenus. Knowledge of the viruses that are being transmitted in the Amazon Basin region of Peru will enable the development of more effective diagnostic assays, more efficient and rapid diagnoses of clinical illnesses caused by these pathogens, risk analysis for military/civilian operations, and development of potential disease control measures.

Vector competence of selected North American Culex and Coquillettidia mosquitoes for West Nile virus.

To control West Nile virus (WNV), it is necessary to know which mosquitoes are able to transmit this virus. Therefore, we evaluated the WNV vector potential of several North American mosquito species. Culex restuans and Cx. salinarius, two species from which WNV was isolated in New York in 2000, were efficient laboratory vectors. Cx. quinquefasciatus and Cx. nigripalpus from Florida were competent but only moderately efficient vectors. Coquillettidia perturbans was an inefficient laboratory vector. As WNV extends its range, exposure of additional mosquito species may alter its epidemiology.

Ochlerotatus j. japonicus in Frederick County, Maryland: discovery, distribution, and vector competence for West Nile virus.

Ochlerotatus japonicus japonicus is reported for the 1st time south of the Mason-Dixon Line, in Frederick County, Maryland. Fifty-seven oviposition trap samples were collected throughout the county between June 30 and August 24, 2000. From 971 larvae reared from the oviposition traps, 5 species were identified: Ochlerotatus triseriatus (45%), Oc. j. japonicus (43%), Aedes albopictus (7%), Culex pipiens (4%), and Toxorynchites ritulus septentrionalis (<1%). Ochlerotatus j. japonicus was found widely distributed over the area sampled. This is the 1st record of Ae. albopictus in the county as well. Vector competence studies indicated that Oc. j. japonicus is an efficient laboratory vector of West Nile (WN) virus. Depending on the viral titer at time of feeding, the estimated transmission rates for Oc. j. japonicus for WN virus were 2-4 times higher than that for Cx. pipiens. Studies of the viral titer in mosquitoes over time showed that titers in the bodies of infected Oc. j. japonicus reached their peak (approximately 10(6.5) plaque-forming units/mosquito) between 7 and 11 days after taking an infectious blood meal, and that virus became detectable in the legs (an indicator of disseminated infection) as early as 3 days after taking an infectious blood meal.

Aedes (Finlaya) japonicus (Diptera: Culicidae), a newly recognized mosquito in the United States: analyses of genetic variation in the United States and putative source populations.

Introduction of potential disease vectors into a new geographic area poses health risks to local human, livestock, and wildlife populations. It is therefore important to gain understanding of the dynamics of these invasions, in particular its sources, modes of spread after the introduction, and vectorial potential. We studied the population genetics of Aedes (Finlaya) japonicus japonicus (Theobald), an Asian mosquito that was recognized for the first time in the United States in 1998. We examined patterns of genetic diversity using random amplified polymorphic DNA and sequences of ND4 of mtDNA by comparing samples from populations spanning the range of this mosquito in Japan (six samples) and the United States (nine samples) as well as specimens intercepted in New Zealand in 1999. We found geographically differentiated populations in Japan, indicating limited gene flow even on small spatial scales. In the United States, we found evidence of significant genetic differentiation between samples from New York, Connecticut, and New Jersey and those from mid-Pennsylvania and Maryland. We were unable to pinpoint the source location(s) in Japan, although some of the U.S. samples are genetically close to samples from south Honshu and western Kyushu. Further studies should include samples from Korean populations. Distinct genetic signatures in U.S. populations undergoing expansion suggest the possibility of local increases in genetic diversity if and where they meet.

Potential North American vectors of West Nile virus.

The outbreak of disease in the New York area in 1999 due to West Nile (WN) virus was the first evidence of the occurrence of this virus in the Americas. To determine potential vectors, more than 15 mosquito species (including Culex pipiens, Cx. nigripalpus, Cx. quinquefasciatus, Cx. salinarius, Aedes albopictus, Ae. vexans, Ochlerotatus japonicus, Oc. sollicitans, Oc. taeniorhynchus, and Oc. triseriatus) from the eastern United States were evaluated for their ability to serve as vectors for the virus isolated from birds collected during the 1999 outbreak in New York. Mosquitoes were allowed to feed on one- to four-day old chickens that had been inoculated with WN virus 1-3 days previously. The mosquitoes were incubated for 12-15 days at 26 degrees C and then allowed to refeed on susceptible chickens and assayed to determine transmission and infection rates. Several container-breeding species (e.g., Ae. albopictus, Oc. atropalpus, and Oc. japonicus) were highly efficient laboratory vectors of WN virus. The Culex species were intermediate in their susceptibility. However, if a disseminated infection developed, all species were able to transmit WN virus by bite. Factors such as population density, feeding preference, longevity, and season of activity also need to be considered in determining the role these species could play in the transmission of WN virus.

Vector competence of Peruvian mosquitoes (Diptera: Culicidae) for epizootic and enzootic strains of Venezuelan equine encephalomyelitis virus.

Mosquitoes collected in the Amazon Basin, near Iquitos, Peru, were evaluated for their susceptibility to epizootic (IAB and IC) and enzootic (ID and IE) strains of Venezuelan equine encephalomyelitis (VEE) virus. After feeding on hamsters with a viremia of approximately 10(8) plaque-forming units of virus per milliliter, Culex (Melanoconion) gnomatus Sallum, Huchings, & Ferreira, Culex (Melanoconion) vomerifer Komp, and Aedes fulvus (Wiedemann) were highly susceptible to infection with all four subtypes of VEE virus (infection rates > or = 87%). Likewise, Psorophora albigenu (Peryassu) and a combination of Mansonia indubitans Dyar & Shannon and Mansonia titillans (Walker) were moderately susceptible to all four strains of VEE virus (infection rates > or = 50%). Although Psorophora cingulata (Fabricius) and Coquillettidia venezuelensis (Theobald) were susceptible to infection with each of the VEE strains, these two species were not efficient transmitters of any of the VEE strains, even after intrathoracic inoculation, indicating the presence of a salivary gland barrier in these species. In contrast to the other species tested, both Culex (Melanoconion) pedroi Sirivanakarn & Belkin and Culex (Culex) coronator Dyar & Knab were nearly refractory to each of the strains of VEE virus tested. Although many of the mosquito species found in this region were competent laboratory vectors of VEE virus, additional studies on biting behavior, mosquito population densities, and vertebrate reservoir hosts of VEE virus are needed to incriminate the principal vector species.

Limited potential for transmission of live dengue virus vaccine candidates by Aedes aegypti and Aedes albopictus.

To evaluate the transmission risk of four live dengue (DEN) vaccine candidates developed by the U.S. Army (DEN-1, 45AZ5 PDK 20; DEN-2, S16803 PDK 50; DEN-3, CH53489 PDK 20; and DEN-4, 341750 PDK 20), we tested 3,010 Aedes aegypti and 1,576 Aedes albopictus mosquitoes blood-fed on 21 volunteers who had been administered one of the four vaccine candidates or the licensed yellow fever (YF) vaccine (17D). We used an indirect immunofluorescence assay (IFA) to detect DEN or YF viral antigen in the heads of mosquitoes. Corresponding to the lack of a detectable viremia among volunteers inoculated 8-13 days previously with live DEN-1 or DEN-2 vaccine candidates, only six mosquitoes developed disseminated infections after feeding on these volunteers. These six mosquitoes included 4 of 247 Ae. albopictus fed on volunteers inoculated with the DEN-1 vaccine candidate and 2 of 528 Ae. aegypti fed on volunteers inoculated with the DEN-2 vaccine candidate. Infection was confirmed in each of these IFA-positive mosquitoes by isolating infectious virus from the mosquito's body in Vero-cell culture. None of the 1,252 or the 969 mosquitoes fed on DEN-3 or DEN-4 recipients, respectively, were infected. Overall, dissemination rates in Ae. albopictus and Ae. aegypti were low. Dissemination rates were 0.5%, 0.3%, < 0.1%, and < 0.1% for the DEN-1 through DEN-4 vaccine candidates, respectively. Because of the observed low dissemination rates, it is unlikely that these vaccine viruses would be transmitted under natural conditions.

Barrier spraying to control the malaria vector Anopheles albimanus: laboratory and field evaluation in the Dominican Republic.

To develop a new strategy for control of exophilic/exophagic malaria vectors which rest on peridomestic foliage, Anopheles albimanus was used for laboratory bioassays of mosquito adulticides applied to various vegetation types. Of the three classes of insecticide tested, the pyrethroid (permethrin at 112 g/ha) showed greater foliar persistence than either the carbamate (bendiocarb at 340 g/ha) or the organophosphorus compound (malathion at 277 g/ha). Field evaluation of barrier spraying against An.albimanus was evaluated in rural villages of southwest Dominican Republic during 1989. The pyrethroid deltamethrin was sprayed aerially for ultra-low volume application at treatment rates of 17-19 g a.i./ha in a radius of 500 m around two villages. An.albimanus were monitored by light-traps and human bait collections at both treated villages, compared with two similar untreated villages, up to 9 nights posttreatment. Densities of female An.albimanus were significantly reduced in the sprayed villages for at least 8 or 9 nights. Further evaluation of barrier spraying is recommended to determine optimal pyrethroid formulations and applications rates, their impact on non-target fauna and efficacy against malaria transmission.

Field evaluation of two formulations of cyfluthrin for control of Ixodes dammini and Amblyomma americanum (Acari: Ixodidae).

Liquid or granular cyfluthrin was applied in the spring and fall to different test plots at a rate of 0.41 (AI) kg/ha to control Ixodes dammini Spielman, Clifford, Piesman & Corwin and Amblyomma americanum (L.). Spring application of liquid cyfluthrin showed the most significant decreases of both species. I. dammini nymphs were reduced 97% at 10 d, 100% at 2 mo, and 100% at 1 yr; and A. americanum adults and nymphs were reduced 91-93% at 10 d and 100% at 2 mo posttreatment. Granular cyfluthrin applied in the spring gave 97 and 87% control of nymphal I. dammini 10 d and 2 mo posttreatment, respectively.

Quantitative evaluation of sampling methods for Ixodes dammini and Amblyomma americanum (Acari: Ixodidae).

Three tick-sampling methods (dry ice-baited tick traps, cloth drags, and ambulatory human host) were evaluated to determine which technique yielded the greatest capture of host-seeking stages of Ixodes dammini Spielman, Clifford, Piesman & Corwin and Amblyomma americanum (L.). The most reliable method, catching more stages and significantly more numbers of I. dammini and A. americanum; was dry ice-baited tick traps. There were no significant differences between the drag and human-host methods for any stage of ticks (I. dammini and A. americanum) collected. The numbers of ticks caught during the study were 5,052 by dry ice-baited tick traps, 199 by cloth drags, and 89 by ambulatory human host.

Comparison of ground and aerial ultra-low volume applications of malathion against Aedes aegypti in Santo Domingo, Dominican Republic.

Efficacy of ground and aerial ultra-low volume (ULV) applications of 91% malathion at 438 ml/ha against Aedes aegypti in the Dominican Republic was evaluated using indoor collections, oviposition trapping and adult sentinel mortality rates. Ground compared to aerial ULV applications in this study were found to have a greater effect on Ae. aegypti when measured by the described sampling techniques. Neither application method provided the level of Ae. aegypti suppression believed necessary for control in the event of a dengue virus epidemic.

Reduction of the Amblyomma americanum (Acari: Ixodidae) population at Fort A.P. Hill, Virginia, by aerial application of diazinon granules: correlation of percentage control with received dose.

Habitat (280 ha) of lone star tick, Amblyomma americanum (L.), was aerially treated with 14% diazinon granules. Concentration of acaricide reaching the forest floor ranged from 0.02 to 1.25 kg (AI)/ha. Greater than 85% control was achieved at study plots receiving the highest dose. A positive correlation was shown between dose received at the forest floor and percentage of control of adults (r = 0.91) and nymphs (r = 0.86).

A new type of collapsible insect-surveillance light trap for sampling Diptera.

The newly designed Army Collapsible Insect Surveillance trap for nocturnal collection of flying Diptera that are attracted by light is described. A polyvinyl chloride construction increases durability, while the capability to collapse from a height of 92.1 cm to 24.1 cm facilitates transportation and storage.