Spatial and temporal distribution of resting female mosquitoes (Diptera: Culicidae) in the coastal plain of North Carolina.

The spatial and temporal distribution of 28 species of female mosquitoes resting in natural (swamp, woods, and swamp-woods ecotone) and human-made (beneath bridges) habitats in blackwater stream-associated ecosystems in Duplin County, N.C., was determined by sampling with a D-Vac vacuum aspirator during 1984 and 1985. Two types of resting distributions were evident. In type one, species including Aedes atlanticus Dyar & Knab, Ae. canadensis (Theobald), Ae. triseriatus (Say), and Psorophora ferox (von Humboldt) rested predominantly on vegetation and were not collected beneath bridges. In type two, species including Anopheles punctipennis (Say), Culex erraticus (Dyar & Knab), Cx. peccator Dyar & Knab, Cx. pipiens L. and Cx. quinquefasciatus Say and their hybrids, Cx. restuans Theobald, Cx. territans Walker, and Uranotaenia sapphirina (Osten Sacken) rested on vegetation, in natural shelters in swamp habitats, and beneath bridges. Differences in the proportion of gravid mosquitoes among habitats were evident for Ae. canadensis, An. punctipennis, Cx. pipiens and Cx. quinquefasciatus and their hybrids, Cx. restuans, Cx. territans, Ps. ferox, and Ur. sapphirina, indicating that gonotrophic condition may influence resting site selection.     

Spatial dispersion of adult mosquitoes (Diptera: Culicidae) in a sylvatic focus of Venezuelan equine encephalitis virus.

We studied the spatial localization of mosquitoes in sylvatic focus of Venezuelan equine encephalitis virus in western Venezuela to identify mosquito species potentially involved in the hypothesized transport of viruses out of enzootic foci. The following criteria were used to identify species with potential for virus export: (1) common in the forest and surrounding area, (2) feeding on a wide range of vertebrates; (3) long dispersal capabilities, and (4) established vectorial competence for enzootic or epizootic VEE viruses. CDC traps baited with light/CO2 were operated for four and 12-h intervals to collect mosquitoes at four stations along two forest/open area transects from September to November 1997. We collected 60,444 mosquitoes belonging to 11 genera and 34 species. The most common species were Aedes serratus (Theobald), Ae. scapularis (Rondani), Ae. fulvus (Wiedmann), Culex nigripalus Theobald, Cx, (Culex) "sp", Cx. mollis Dyar & Knab, Cx. spissipes (Theobald), Cx. pedroi Sirivanakarn and Belkin, Psorophora ferox (Humboldt), Ps. albipes (Theobald), and Ps. cingulata (F.). Very few mosquitoes were captured during the (day in the open area outside the forest, suggesting that any virus export from the forest may occur at night. The following mosquitoes seemed to be mostly restricted to the forest habitat: Ae. serratus, Ps. ferox, Ps. albipes, sabethines, Cx. spissipes, Cx. pedroi, Cx. dunni Dyar, and Ae. fulvus. The main species implicated its potential virus export were Cx. nigripalpus, Ae. scapularis, and Mansonia titillans (Walker).     

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.     

Targeted trapping of mosquito vectors in the Chesapeake Bay area of Maryland

Most adult mosquito surveillance in Maryland is performed using dry ice-baited or unbaited Centers for Disease Control (CDC) miniature light traps suspended approximately 1.5 m above the ground. However, standardized trapping methods may miss mosquito species involved in disease transmission cycles. During a 2-yr study, the effectiveness of the olfactory attractant 1-octen-3-ol alone and in combination with carbon dioxide was evaluated for collecting mosquito vector species. In addition, trap heights were examined to determine the optimal vertical placement to target various species. We evaluated the results during the second year by targeting selected species by using various habitat- height-bait combinations. Although Culex erraticus Dyar & Knab and Anopheles quadrimaculatus Say were not successfully targeted, Culex salinarius Coquillett, Aedes vexans Meigen, Anopheles bradleyi/crucians King, Coquillettidia perturbans Walker, Aedes sollicitans Walker, and Aedes taeniorhynchus Wiedemann were preferentially captured using targeted trapping schemes.     

Susceptibility of Peruvian mosquitoes to eastern equine encephalitis virus

Mosquitoes were collected in the Amazon Basin, near Iquitos, Peru, and used in experimental studies to evaluate their susceptibility to strains of eastern equine encephalitis virus (EEEV) that were isolated from mosquitoes captured within 20 km of Iquitos. When fed on hamsters or chickens with a viremia of 4105 plaque-forming units (PFU) of EEEV/ml, Culex pedroi Sirivanakarn and Belkin, Aedesfulvus (Wiedemann), Psorophora albigenu (Peryassu), and Psorophoraferox (Von Humboldt) were susceptible to infection, whereas none of the Aedes serratus (Theobald), Culex vomerifer Komp, Culex gnomatos Sallum, Huchings, and Ferreira, Culex portesi Senevet and Abonnenc, or Culex coronator Dyar and Knab became infected, even though they fed on the same viremic blood sources. When these mosquito species fed on animals with viremias of approximately 10(8) PFU/ml, Cx. pedroi, Ae.II (Brazil-Peru) and a lineage III (Argentina-Panama) isolate of EEEV. This study, combined with the repeated isolation of strains of EEEV from Cx. pedroi captured in the Amazon Basin region of Peru, suggests that Cx. pedroi may be the primary enzootic vector of EEEV in this region.     

West Nile virus infection in mosquitoes in the mid-south USA, 2002-2005

West Nile virus (family Flaviviridae, genus Flavivirus, WNV) was first detected in the Tennessee Valley and in Alabama in August 2001. In summer 2002, intensive viral activity was seen, but in subsequent years, viral activity settled into an enzootic pattern. Here, we report an analysis of viral activity in the mosquito fauna in the Mid-South from 2002 (the first year viral activity was detected in mosquitoes) through 2005. Eight mosquito species were infected with WNV during 2002. However, viral activity was only detected in four species--Culex salinarius Coquillett, Culex erraticus Dyar & Knab, Coquillettidia perturbans Walker, and Aedes vexans Meigen--in multiple years. The greatest number of positive pools was in Cx. erraticus and Cx. salinarius. Despite being specifically targeted for collection, Aedes albopictus Skuse was only found to be infected during the epiornitic year (2002), suggesting that under enzootic transmission conditions its role as a bridge vector in the region may not be significant. Virus-positive pools of Cx. erraticus were identified from winter-resting and early season dry ice-baited trap collections in 2005, implicating this species in WNV overwintering in Alabama. Molecular analysis of individuals initially identified as members of the Culex pipiens L. complex suggested that alleles characteristic of Cx. pipiens predominated in mosquitoes collected in Huntsville, AL, whereas alleles in the Auburn, AL, population were predominately characteristic of Culex quinquefaciatus Say. The southern boundary of the overlap zone of the two species seems to be located primarily between Huntsville and Auburn, a distance of 350 km.     

Molecular phylogeny and evolutionary relationships among mosquitoes (Diptera: Culicidae) from the northeastern United States based on small subunit ribosomal DNA (18S rDNA) sequences

The phylogenetic relationships of Culicidae native to the northeastern United States were investigated by analyzing small subunit ribosomal DNA (18S rDNA) sequences obtained from 39 species representing nine genera. Molecular phylogenies were consistent with traditional classifications based on morphological characters except for the placements of Psorophora Robineau-Desvoidy and Uranotaenia Lynch Arribalzaga. In our analyses, 1) Anopheles Meigen was strongly supported as the sister taxon to the remaining Culicidae; 2) Toxorhynchites Theobald was represented as a distinct monophyletic sister group to the Culicinae; 3) Psorophora formed a basal clade to Culiseta Felt, Coquillettidia Dyar, and Culex L. but also was shown as a sister taxon to Aedes Meigen and Ochlerotatus Lynch Arribalzaga; 4) Coquillettidia perturbans (Walker) seems to be a sister group to Culiseta; 5) placement of Uranotaenia was inconclusive and seemed to be either a sister group to the Aedes and Ochlerotatus or a basal taxon to all other culicines; and 6) Aedes and Ochlerotatus formed two separate and distinct clades, providing phylogenetic data consistent with the recent elevation of Ochlerotatus to the generic level as proposed by Reinert (2000). The utility of 18S rDNA for evaluating phylogenetic and evolutionary relationships among mosquito taxa was demonstrated at the genus and species levels. To our knowledge, this study represents the first molecular-based phylogenetic study of mosquito species occurring within this geographic region of North America and contains the largest number of species that have been examined among the genera Aedes and Ochlerotatus.     

Vector competence of three Venezuelan mosquitoes (Diptera: Culicidae) for an epizootic IC strain of Venezuelan equine encephalitis virus.

Experimental studies were undertaken to evaluate the vector competence of selected mosquito species [Aedes taeniorhynchus (Wiedemann), Culex declarator Dyar and Knab, and Mansonia titillans (Walker)] from northwestern Venezuela for the epizootic (IC) strain of Venezuelan equine encephalitis (VEE) virus that was responsible for the 1995 outbreak of VEE in this area. Ae. taeniorhynchus was highly susceptible to infection (94% of 35), and 89% had a disseminated infection. Virus-exposed Ae. taeniorhynchus that refed on susceptible hamsters readily transmitted virus, confirming that this species was an efficient vector of VEE virus. In contrast, only 1 of 28 (4%) Cx. declarator was infected, and that individual did not develop a disseminated infection. Ma. titillans was moderately susceptible (3 of 8 infected, 38%), and 2 (25%) of these had a disseminated infection. These data indicate that Ae. taeniorhynchus was an important epizootic vector during the 1995 VEE outbreak in Columbia and Venezuela.     

Host specificity of Amblyospora connecticus (Microsporida: Amblyosporidae), a polymorphic microsporidian parasite of Aedes cantator (Diptera: Culicidae)

Amblyospora connecticus Andreadis, a microsporidian parasite of the brown saltmarsh mosquito, Aedes cantator (Coquillett), was successfully transmitted to Ae. atropalpus (Coquillett), Ae. epactius Dyar and Knab, Ae. sierrensis (Ludlow), and Ae. triseriatus (Say) following larval exposure to spores from the intermediate copepod host, Acanthocyclops vernalis (Fischer). The microsporidium underwent normal vegetative growth and multiplication in fat body, muscle, and Malpighian tubule tissues of all four alternate mosquito hosts but produced spores only in female Ae. epactius. However, in no alternate host was A. connecticus able to infect the ovaries and complete its life cycle via transovarial transmission, thus indicating a high degree of specificity for Ae. cantator. Fifteen other mosquito species in Aedes, Anopheles, Culex, Culiseta, and Psorophora also were tested and found to be nonsusceptible.     

Estimation of dispersal distances of Culex erraticus in a focus of eastern equine encephalitis virus in the southeastern United States

Patterns of mosquito dispersal are important for predicting the risk of transmission of mosquito-borne pathogens to vertebrate hosts. We studied dispersal behavior of Culex erraticus (Dyar & Knab), a potentially significant vector of eastern equine encephalitis virus (EEEV) that is often associated with foci of this pathogen in the southeastern United States. Using data on the relative density of resting adult female Cx. erraticus around known emergence sites in Tuskegee National Forest, Alabama, we developed a model for the exponential decay of the relative density of adult mosquitoes with distance from larval habitats through parameterization of dispersal kernels. The mean and 99th percentile of dispersal distance for Cx. erraticus estimated from this model were 0.97 and 3.21 km per gonotrophic cycle, respectively. Parameterized dispersal kernels and estimates of the upper percentiles of dispersal distance of this species can potentially be used to predict EEEV infection risk in areas surrounding the Tuskegee National Forest focus in the event of an EEEV outbreak. The model that we develop for estimating the dispersal distance of Cx. erraticus from collections of adult mosquitoes could be applicable to other mosquito species that emerge from discrete larval sites.     

Isolation and sequence analysis of Culex flavivirus from Culex interrogator and Culex quinquefasciatus in the Yucatan Peninsula of Mexico

Previously, we reported a high prevalence of Culex flavivirus (CxFV) in Culex quinquefasciatus (Say) in the Yucatan Peninsula of Mexico. To determine whether other Culex spp. mosquitoes in this region are susceptible to natural CxFV infection, Cx. bahamensis (Dyar and Knab), Cx. coronator (Dyar and Knab), Cx. interrogator (Dyar and Knab), Cx. nigripalpus (Theobald) and Cx. opisthopus (Komp) in the Yucatan Peninsula of Mexico were tested for CxFV. Two pools of Cx. interrogator were positive. The envelope protein genes of these isolates and 16 isolates from Cx. quinquefasciatus were sequenced and shown to have ≥99.2% nucleotide identity. These data suggest that there is limited genetic diversity among CxFV isolates in the Yucatan Peninsula of Mexico.     

Developing models for the forage ratios of Culiseta melanura and Culex erraticus using species characteristics for avian hosts

Studies of mosquito preferences for avian hosts have found that some bird species are at greater risk than others of being fed upon by mosquitoes. The ecological factors that determine this interspecific variation in avian host use by mosquitoes have been little studied, despite the possibility that such variation may influence spatial and temporal patterns of the occurrence of mosquito-borne pathogens. Our objective was to identify ecological variables associated with the avian host forage ratios estimated from a previous study of mosquito feeding patterns in Tuskegee National Forest, AL. We used species' characteristics derived from the literature to develop multiple linear regression models for the forage ratios of Culiseta melanura (Coquillett) and Culex erraticus (Dyar & Knab) for avian hosts. We found that habitat-edge association and body mass of avian host species were the best predictors of forage ratios of Cx. erraticus for avian hosts. Although no avian host traits were inferred to be strong predictors of forage ratios of Cs. melanura, body mass had the greatest importance weight among those considered. Our results suggest that characteristics of avian hosts may predict their levels of use by some mosquito species.     

Eastern equine encephalitis in Tennessee: 2002-2008

Human and equine outbreaks caused by eastern equine encephalomyelitis virus (EEEV) typically occur in North America adjacent to coastal wetlands associated with the presence of Culiseta melanura (Coquillet) mosquitoes. Eastern equine encephalomyelitis (EEE) is an emerging disease in Tennessee, as the first records of equine disease began in 2002. In 2006 we trapped and tested mosquitoes for EEEV at hardwood swamps in western Tennessee that were at the epicenter of a multi-equine outbreak in fall of 2005. Additionally, the Tennessee Valley Authority tested mosquito pools collected in Tennessee swamps from 2000 to 2007 for the presence of arboviruses. Two pools of EEEV positive Culex erraticus (Dyer and Knab) mosquitoes were found (one each in 2003 and 2004) in a county adjacent to where the 2005 outbreak occurred. In 2008, another EEE outbreak involving multiple horses occurred in West Tennessee. A brain specimen was collected from a horse during this outbreak and the first isolate of EEEV from Tennessee was obtained. In total, 74,531 mosquitoes collected from 2000 to 2008 were tested via polymerase chain reaction and VecTest for EEEV. The traditional enzootic vector, Cs. melanura, was found in low numbers at all collection sites. Cx. erraticus, however, was consistently found in high numbers and was the only mosquito species in which EEEV was detected. We suggest that EEE transmission may be maintained by Cx. erraticus in a nontraditional cycle. We discuss the importance of a nontraditional cycle from the perspective of EEEV adaptation and emergence.     

Potential for North American mosquitoes (Diptera: Culicidae) to transmit rift valley fever virus

To determine which arthropods should be targeted for control should Rift Valley fever virus (RVFV) be detected in North America, we evaluated Culex erraticus (Dyar and Knab), Culex erythrothorax Dyar, Culex nigripalpus Theobald, Culex pipiens L., Culex quinquefasciatus Say, Culex tarsalis Coquillett, Aedes dorsalis (Wiedemann), Aedes vexans (Meigen), Anopheles quadrimaculatus Say, and Culicoides sonorensis Wirth and Jones from the western, midwestern, and southern United States for their ability to transmit RVFV. Female mosquitoes were allowed to feed on adult hamsters inoculated with RVFV, after which engorged mosquitoes were incubated for 7-21 d at 260C, then allowed to refeed on susceptible hamsters, and tested to determine infection, dissemination, and transmission rates. Other specimens were inoculated intrathoracically, held for 7 d, and then allowed to feed on a susceptible hamster to check for a salivary gland barrier. When exposed to hamsters with viremias > or =10(8.8) plaque-forming units/ml blood, Cx. tarsalis transmitted RVFV efficiently (infection rate = 93%, dissemination rate = 56%, and estimated transmission rate = 52%). In contrast, when exposed to the same virus dose, none of the other species tested transmitted RVFV efficiently. Estimated transmission rates for Cx. erythrothorax, Cx. pipiens, Cx. erraticus, and Ae. dorsalis were 10, 8, 4, and 2%, respectively, and for the remaining species were < or = 1%. With the exception of Cx. tarsalis and Cx. pipiens, all species tested had moderate to major salivary gland barriers. None of the C. sonorensis became infected and none of the An. quadrimaculatus tested transmitted RVFV by bite, even after intrathoracic inoculation, indicating that these species would not be competent vectors of RVFV. Although Ae. vexans from Florida and Louisiana were relatively efficient vectors of RVFV, specimens of this species captured in Colorado or California were virtually incompetent, illustrating the need to evaluate local population for their ability to transmit a pathogen. In addition to laboratory vector competence, factors such as seasonal density, host feeding preference, longevity, and foraging behavior should be considered when determining the potential role that these species could play in RVFV transmission.     

Isolation of Rift Valley fever virus from mosquitoes (Diptera: Culicidae) collected during an outbreak in domestic animals in Kenya.

During an outbreak of Rift Valley fever (RVF) in livestock near Lake Naivasha, Rift Valley Province, Kenya, 61,347 mosquitoes (1,287 pools) collected in CO2-baited light traps yielded seven viral isolates. Five isolates of RVF virus were recovered from 18,831 Culex zombaensis Theobald and one from 14,439 Mansonia africana (Theobald). One isolate of a Bunyamwera group virus was recovered from 1,175 Aedes quasiunivittatus (Theobald).     

Comparison of specific composition and mosquitoes population dynamics in two villages of the centre of Côte-d'Ivoire, with and without irrigated rice growing area

Entomological surveys were undertaken between October 1994 and August 1995 and from April to August 1995 in the villages of Kafiné and Pékaha, respectively These villages are located in the centre of C?te-d'Ivoire in a humid savannah area in the prefecture of Niakaramandougou. Rice plots are located to about 500 meters from the first habitations of Kafiné. They also form an enclosure around the village causing a great culicidian nuisance. On the other hand in the village of Pékaha, situated at 8 km from the enclosure, one can find a non-developed low ground area with swamps according to the season which has caused the multiplication of anthropophilic Culicidae. Five genera of mosquito were caught in the two villages: Anopheles, Aedes, Culex, Mansonia and Uranotaenia. The genus Anopheles is by far the most predominant in Kafiné with 84.3% (n=17556). Mansonia, Culex and Aedes respectively constitute 12%, 3% and 0.01% of the total anthropophilic mosquitoes. In the contrary the genus Mansonia was the most predominant in Pékaha with 78.2% (n=6642). Anopheles, Aedes and Culex respectively represent 13.2%, 8.3% and 0.1% of the total number of mosquitoes caught during the surveys. If in the village of Kafiné, the culicidian nuisance is mainly due to anopheles mosquitoes (84.3%) and particularly to An. gambiae s.s. (70.6%), in the village of Pékaha, dwellers suffer more from Mansonia (78.2%) bites than those of Anopheles (13.2%).