Vector competence of Mexican and Honduran mosquitoes (Diptera: Culicidae) for enzootic (IE) and epizootic (IC) strains of Venezuelan equine encephalomyelitis virus

Experimental studies evaluated the vector competence of Ochlerotatus taeniorhynchus (Wiedemann), Culex cancer Theobald, Culex pseudes (Dyar and Knab), Culex taeniopus Dyar and Knab, and a Culex (Culex) species, probably Culex quinquefasciatus Say, and Culex nigripalpus Theobald from Chiapas, Mexico, and Tocoa, Honduras, for epizootic (IC) and enzootic (IE) strains of Venezuelan equine encephalomyelitis (VEE) virus. Culex pseudes was highly susceptible to infection with both the IC and IE strains of VEE (infection rates >78%). Patterns of susceptibility to VEE were similar for Oc. taeniorhynchus collected in Mexico and Honduras. Although Oc. taeniorhynchus was highly susceptible to the epizootic IC strains (infection rates > or = 95%, n = 190), this species was less susceptible to the enzootic IE strain (infection rates < or = 35%, n = 233). The Culex (Culex) species were refractory to both subtypes of VEE, and none of 166 contained evidence of a disseminated infection. Virus-exposed Cx. pseudes that refed on susceptible hamsters readily transmitted virus, confirming that this species was an efficient vector of VEE. Although Oc. taeniorhynchus that fed on hamsters infected with the epizootic IC strain transmitted VEE efficiently, only one of six of those with a disseminated infection with the enzootic IE virus that fed on hamsters transmitted virus by bite. These data indicate that Cx. pseudes is an efficient laboratory vector of both epizootic and enzootic strains of VEE and that Oc. taeniorhynchus could be an important vector of epizootic subtypes of VEE.     

Transmission of Venezuelan equine encephalomyelitis virus by Aedes sollicitans and Aedes taeniorhynchus (Diptera: Culicidae).

Experimental studies compared the vector competence of Aedes sollicitans (Skuse) and Ae. taeniorhynchus (Wiedemann) collected on Assateague Island, Va., for an epizootic strain (Trinidad donkey) of Venezuelan equine encephalomyelitis (VEE) virus. Infection rates were significantly higher in Ae. sollicitans (101/107, 94%) than in Ae. taeniorhynchus (103/175, 59%), even though both species fed concurrently on the same infected hamsters. Similarly, dissemination and transmission rates were significantly higher in the Ae. sollicitans population tested. Although both Ae. taeniorhynchus and Ae. sollicitans are natural vectors of VEE virus, the latter species should be considered a more efficient vector of VEE epizootic strains, based on its greater susceptibility to infection and higher transmission rates.     

Brugia malayi microfilariae (Nematoda: Filaridae) enhance the infectivity of Venezuelan equine encephalitis virus to Aedes mosquitoes (Diptera: Culicidae)

We examined the potentially conflicting effects that microfilarial (MF) enhancement of viral infectivity and MF-induced mortality in mosquitoes have on the vectorial capacity of Aedes aegypti (L.), Aedes triseriatus (Say), and Aedes taeniorhynchus (Wiedemann) for Venezuelan equine encephalitis virus (VEE) when mosquitoes feed on gerbils co-infected with Brugia malayi (Buckley). Groups of mosquitoes were fed on gerbils that were either dually infected (VEE plus B. malayi MF) or singly infected (VEE only). Mosquito mortality was recorded daily, and 5-8 d later, surviving mosquitoes were assayed for disseminated viral infection. The contrasting effects of MF enhancement and MF-induced mortality differed among mosquito species and were determined by the nature and consequences of MF penetration through the mosquito midgut, but not to differences in mosquito susceptibilities to parenterally introduced virus. In Ae. aegypti, MF-induced mortality was high and tended to eliminate any significant effect of MF enhancement. In Ae. triseriatus, MF-induced mortality was low, and feeding on dually infected hosts resulted in 9 times as many mosquitoes with disseminated viral infections as did feeding on singly-infected hosts. In Ae. taeniorhynchus, MF-induced mortality was extremely high, yet under our experimental conditions, feeding on a dually infected hosts resulted in nearly 30 times as many disseminated infections as did feeding on singly infected hosts. The final outcome on vectorial capacity depended on the specific combination of MF, virus, and mosquito species involved. Therefore, future efforts toward understanding MF enhancement should be directed toward mosquito-virus-parasite species combinations that occur together in nature.     

Transmission of Venezuelan equine encephalomyelitis virus by strains of Aedes albopictus (Diptera: Culicidae) collected in North and South America

Experimental studies were undertaken to ascertain the vector potential of North American (Houston and Alsace) and South American (Sao Paulo and Santa Teresa) strains of Aedes albopictus (Skuse) for an epizootic (Trinidad donkey) strain of Venezuelan equine encephalomyelitis (VEE) virus. Infection rates were similar in all four strains of Ae. albopictus tested after ingestion of VEE virus from a viremic hamster. Virus disseminated from the midgut to the hemocoel in about 80% of infected mosquitoes, regardless of the dose ingested (10(4.6) to 10(5.7) plaque-forming units per mosquito) or the time of extrinsic incubation (7-35 d). Although all four strains of this mosquito transmitted VEE virus by bite to hamsters, transmission rates were significantly higher for the South American strains (24%, 40 of 170) than for the North American strains (5%, 9 of 165). Although VEE virus has never been isolated from Ae. albopictus, the introduction of this species into the Americas may allow it to serve as an amplification vector in areas where epizootic strains of VEE are found or introduced.     

Susceptibility of Ochlerotatus taeniorhynchus (Diptera: Culicidae) to infection with epizootic (subtype IC) and enzootic (subtype ID) Venezuelan equine encephalitis viruses: evidence for epizootic strain adaptation

To test the hypothesis that adaptation to epizootic mosquito vectors mediates emergence of Venezuelan equine encephalitis virus (VEEV) from enzootic progenitors, experimental infection studies were conducted to determine the susceptibility of Ochlerotatus taeniorhynchus (Wiedemann) to epizootic and enzootic strains. Artificial blood meals containing epizootic subtype IC strains isolated during the 1962-1964, 1992-1993, and 1995 Venezuelan/Colombian epizootics and closely related Venezuelan enzootic subtype ID strains were used to compare infectivity and transmission potential. Their greater infectivity and replication suggested that adaptation of epizootic strains to Oc. taeniorhynchus may have enhanced epizootic transmission during the 1962-1964 and 1995 IC coastal epizootics. However, strains from the small 1992-1993 Venezuelan outbreak that did not extend to coastal regions do not seem to infect this species better than closely related subtype ID strains. Adaptation of VEEV to epizootic vectors such as Oc. taeniorhynchus mosquitoes may be a determinant of some but not all VEE emergence events and may influence spread into coastal regions.     

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.     

Experimental transmission of Venezuelan equine encephalomyelitis virus by a strain of Aedes albopictus (Diptera: Culicidae) from New Orleans, Louisiana.

Experimental studies were undertaken to ascertain the vector competence of a strain of Aedes albopictus (Skuse) collected in New Orleans, LA, (Gentilly strain) for an epizootic (Trinidad donkey) strain of Venezuelan equine encephalomyelitis (VEE) virus. This strain of Ae. albopictus was significantly more susceptible to infection with VEE virus than were any of the four strains tested previously, including two from North America and two from South America. Likewise, dissemination (148 of 180) (82%) and transmission (40 of 88) (45%) rates were significantly higher in the Gentilly strain than in any of the strains previously tested. Analysis of the results of the present study along with those of a previous study with a second alphavirus, chikungunya (CHIK) virus, indicated that, although all three strains of Ae. albopictus tested were more susceptible to VEE virus than to CHIK virus, susceptibility to infection and dissemination with one alphavirus appeared to be directly related to susceptibility to infection and dissemination with the other virus and may indicate shared receptor sites for these two alphaviruses in Ae. albopictus.     

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).     

Genetic evidence that epizootic Venezuelan equine encephalitis (VEE) viruses may have evolved from enzootic VEE subtype I-D virus.

An important question pertaining to the natural history of Venezuelan equine encephalitis (VEE) virus concerns the source of epizootic, equine-virulent strains. An endemic source of epizootic virus has not been identified, despite intensive surveillance. One of the theories of epizootic strain origin is that epizootic VEE viruses evolve from enzootic strains. Likely enzootic sources of VEE virus occur in Colombia and Venezuela where many of the epizootic outbreaks of VEE have occurred. We have determined the nucleotide sequences of the entire genomes of epizootic VEE subtype I-C virus, strain P676, isolated in Venezuela, and of enzootic VEE subtype I-D virus, strain 3880, isolated in Panama. VEE subtype I-D viruses are maintained in enzootic foci in Panama, Colombia, and Venezuela. The genomes of P676 and 3880 viruses differ from that of VEE subtype I-AB virus, strain Trinidad donkey (TRD), by 417 (3.6%) and 619 (5.4%) nucleotides, respectively. The translated regions of P676 and 3880 genomes differ from those of TRD virus by 54 (1.4%) and 66 (1.8%) amino acids, respectively. This study and the oligonucleotide fingerprint analyses of South American I-C and I-D viruses (Rico-Hesse, Roehrig, Trent, and Dickerman, 1988, Am. J. Trop. Med. Hyg. 38, 187-194) provide the most conclusive evidence to date suggesting that equine-virulent strains of VEE virus arise naturally from minor variants present in populations of I-D VEE virus maintained in enzootic foci in northern South America.     

Vector competence of rural and urban strains of Aedes (Stegomyia) albopictus (Diptera: Culicidae) from São Paulo State, Brazil for IC, ID, and IF subtypes of Venezuelan equine encephalitis virus

Aedes albopictus (Skuse) is an Asiatic mosquito species that has spread and colonized all continents except Antarctica. It has major public health importance because it is a potential vector of several pathogens. The objectives of our study were to analyze the vector competence of urban and rural strains of Ae. albopictus from S?o Paulo State (Brazil) for Venezuelan equine encephalitis virus (VEE) subtypes IC, ID, and IF, and to evaluate the effect of infection with subtype IC of VEE on mosquito longevity. Both mosquito strains were susceptible to subtypes IC and ID, but the infection rate for subtype IF was low. Infection and transmission rates of Ae. albopictus for subtype IC were similar to those reported for Ochlerotatus taeniorhynchus (Wiedemann). The high infection, dissemination, and transmission rates for subtype ID reported for Oc. fulvus (Wiedemann) and Culex (Melanoconion) spp. are comparable with those found in this study. We found significant differences in the susceptibility to subtype IC between rural and urban populations of S?o Paulo. Significant survival rate differences were observed between uninfected and infected mosquitoes, but there were no differences in survival between rural and urban mosquito strains.     

Vector competence of North American mosquitoes (Diptera: Culicidae) for West Nile virus

We evaluated the potential for several North American mosquito species to transmit the newly introduced West Nile (WN) virus. Mosquitoes collected in the New York City metropolitan area during the recent WN virus outbreak, at the Assateague Island Wildlife Refuge, VA, or from established colonies were allowed to feed on chickens infected with WN virus isolated from a crow that died during the 1999 outbreak. These mosquitoes were tested approximately 2 wk later to determine infection, dissemination, and transmission rates. Aedes albopictus (Skuse), Aedes atropalpus (Coquillett), and Aedes japonicus (Theobald) were highly susceptible to infection, and nearly all individuals with a disseminated infection transmitted virus by bite. Culex pipiens L. and Aedes sollicitans (Walker) were moderately susceptible. In contrast, Aedes vexans (Meigen), Aedes aegypti (L.), and Aedes taeniorhynchus (Wiedemann) were relatively refractory to infection, but individual mosquitoes inoculated with WN virus did transmit virus by bite. Infected female Cx. pipiens transmitted WN virus to one of 1,618 F1 progeny, indicating the potential for vertical transmission of this virus. In addition to laboratory vector competence, host-feeding preferences, relative abundance, and season of activity also determine the role that these species could play in transmitting WN virus.     

Transmission of western equine encephalomyelitis virus by Argentine Aedes albifasciatus (Diptera: Culicidae).

Aedes albifasciatus (Macquart) has been considered a potential vector of western equine encephalitis (WEE) virus in Argentina because it has been found naturally infected, it is susceptible to infection per os, and its distribution and feeding habits are compatible with those of an epizootic and epidemic vector. Ae. albifasciatus females collected in the vicinity of Cordoba, Argentina, were highly susceptible to WEE virus perorally (ID50 less than 0.5 Vero cell plaque-forming units) when fed on viremic chicks. Virus transmission trials were done 9 to 16 d after infection by feeding potentially infected mosquitoes individually on chicks. Among 31 mosquitoes that survived the incubation period, six refed on susceptible chicks. Virus transmission by bite was demonstrated by five of six (83%) mosquitoes. Therefore, the final piece of evidence incriminating Ae. albifasciatus as a natural vector of WEE virus has been obtained. This is the first mosquito species to be incriminated as a vector of WEE virus in South America.     

Venezuelan equine encephalomyelitis virus infection in and transmission by the tick Amblyomma cajennense (Arachnida: Ixodidae)

To assess a possible role of ticks as the maintenance host for epizootic strains of Venezuelan equine encephalomyelitis (VEE) virus, laboratory experiments were conducted to determine if ticks could become infected, maintain, and transmit the virus. Larval and nymphal Amblyomma cajennense (F.) and larval Dermacentor nitens Neumann ticks were exposed to epizootic VEE virus (Trinidad donkey strain) by allowing them to feed on viremic guinea pigs (strain 13). In A. cajennense, transstadial transmission was observed from larvae to nymphs and adults. Horizontal viral transmission to a mammalian host was accomplished by nymphs. Infection rates in nymphs and adults were 2% (42/2,750) and 4% (9/244), respectively, afer ingestion of virus as larvae. Virus was detected in A. cajennense adult ticks for up to 171 d after infection in the larval stage. A cajennense, exposed as nymphs, ingested virus but did not become infected (0/164 after 10 d after taking an infective bloodmeal). No virus was detected in D. nitens 7 d after exposure. These findings suggest that A. cajennense potentially could be involved in an interepizootic maintenance cycle of epizootic VEE viral strains.     

Ecological characterization of the aquatic habitats of mosquitoes (Diptera: Culicidae) in enzootic foci of Venezuelan equine encephalitis virus in western Venezuela

We studied the aquatic mosquito habitats in and around enzootic foci of Venezuelan Equine Encephalitis virus (VEE) in western Venezuela. Specimens were sampled for 5 mo in three types of vegetation: tall lowland tropical forests, short inundated/secondary growth forests, and pastures/herbaceous vegetation around forests. Ground pools, flooded pastures, swamps, ponds, and canals predominated. We used a multivariate statistical approach to quantitatively assess the relationships of mosquito species with broad categories of the landscape, and with environmental variables within each aquatic habitat. Twenty-four mosquito species in the genera Aedes, Psorophora, Culex, Mansonia, and Uranotaenia were collected. Species richness was higher in the tall forests than in other types of vegetation. Discriminant Function Analysis showed a strong association between landscape category and mosquito species assemblage and identified Culex erraticus Dyar & Knab and Mansonia titillans Walker as indicator species of open areas, and Aedes serratus Theobald as an indicator of tall forests. M. titillans, Uranotaenia geometrica Theobald, Cx. erraticus, and Culex dunni Dyar were associated with unshaded, warm, vegetated waters in flooded pastures and swamps, whereas Ae. serratus, Aedes fulvus (Wiedemann), Psorophora albipes Theobald, Psorophora ferox (Humboldt), Culex caudelli Dyar & Knab, and Culex pedroi Sirivanakarn & Belkin were associated with small, shaded ground pools within the tall forests. Culex coronator Dyar & Knab was associated with partially exposed sites within short forests. These results allowed us to interpret better our previous studies on mosquito adult spread in the study area and their possible role as VEEV disseminators.     

Human Venezuelan equine encephalitis virus infection and diabetes in Zulia State, Venezuela

Venezuelan equine encephalitis (VEE) virus has been implicated as producing alterations in glucose metabolism in animals. We performed oral glucose tolerance tests and measured serum immunoreactive insulin responses in 13 patients who were infected by VEE virus during an epidemic in 1969, in Zulia State, Venezuela. No significant alterations in the glucose tolerance test were found. Sera of 86 diabetic outpatients and 98 control individuals with normal glycemia at a local hospital were tested for antibodies to VEE virus by hemagglutination inhibition. No statistically significant difference was found between the two groups; 10.4% of the diabetic patients had detectable antibodies against VEE virus, compared to 7.1% of controls. Seventy-three percent of the diabetics with antibodies were individuals over 40 yr old, whose diabetes could be catalogued as insulin independent. The results of these studies indicate no relationship of VEE virus infection to subsequent diabetes.     

Comparative potential of Aedes triseriatus, Aedes albopictus, and Aedes aegypti (Diptera: Culicidae) to transovarially transmit La Crosse virus

Aedes triseriatus (Say) (Diptera: Culicidae), the major vector of La Crosse (LAC) virus, efficiently transmits LAC virus both horizontally and transovarially. We compared the vector competence and transovarial transmission ability of Ae. triseriatus, Aedes albopictus Skuse, and Aedes aegypti (L.) for LAC virus. Ae. triseriatus and Ae. albopictus were significantly more susceptible to oral infection with LAC virus than Ae. aegypti. The three species also differed in oral and disseminated infection rates (DIRs). Transovarial transmission (TOT) rates and filial infection rates (FIRs) were greater for Ae. triseriatus than either Ae. albopictus or Ae. aegypti. These measures were integrated into a single numerical score, the transmission amplification potential (TAP) for each species. Differences in TAP scores were due mainly to the differences in DIRs and FIRs among these mosquitoes. Although the TAP score for Ae. albopictus was lower than that of Ae. triseriatus, it was 10-fold greater than that for Ae. aegypti.     

Susceptibility of Psorophora confinnis (Diptera: Culicidae) to infection with epizootic (subtype IC) and enzootic (subtype ID) Venezuelan Equine encephalitis viruses

To test the hypothesis that adaptation to epizootic mosquito vectors mediates the emergence of Venezuelan equine encephalitis virus (family Togaviridae, genus Alphavirus, VEEV) from enzootic progenitors, the susceptibility of the epizootic vector Psorophora confinnis (Lynch-Arribalzaga) to epizootic versus enzootic strains was evaluated. Artificial bloodmeals containing subtype IC strains isolated during the 1962-1964, 1992-1993, and 1995 Venezuelan/Colombian epizootics and closely related Venezuelan enzootic subtype ID strains were used to compare mosquito infectivity and transmission potential. Strains from the smaller 1992-1993 epizootic showed lower or equal infectivity and replication compared with enzootic viruses and to strains isolated during the larger 1962-1964 and 1995 epizootics. These experiments failed to provide evidence that Ps. confinnis selects for epizootic VEEV viruses with higher infectivity, as has been shown for Aedes (Ochlerotatus) taeniorhynchus (Wiedemann). Nonetheless, its high susceptibility, abundance in enzootic and epizootic regions, and feeding behavior suggest that Ps. confinnis is an important bridge vector for both enzootic and epizootic VEEV.     

Replication and dissemination of La Crosse virus in the competent vector Aedes triseriatus and the incompetent vector Aedes hendersoni and evidence for transovarial transmission by Aedes hendersoni (Diptera: Culicidae)

The time course and pattern of the replication and dissemination of La Crosse virus was studied in orally infected Aedes triseriatus (Say) and Ae. hendersoni Cockerell. Development of La Crosse virus was approximately the same in both species when plaque assay titers of intact mosquitos or dissected tissues were compared. The mosquitoes were equally susceptible to infection; all Ae. hendersoni and 99% of the Ae. triseriatus tested showed detectable midgut infections. Virus was first detected in hemolymph, salivary glands, and ovaries 10-13 d after infection in both species. The pattern of infection suggests virus dissemination beyond the midgut to be via the hemolymph. By 21 d after infection, 100% (10 of 10) of Ae. triseriatus and 70% (7 of 10) of Ae. hendersoni had infected salivary glands, and the geometric mean titer of Ae. hendersoni salivary glands was 10 times higher than the geometric mean titer of those of Ae. triseriatus, However, when tested for transmission 22 d after infection by refeeding on suckling mice, only 9% (2 of 22) of the Ae. hendersoni with disseminated infections transmitted virus versus 71% (12 of 17) of the Ae. triseriatus. A salivary gland escape barrier was shown to be primarily responsible for the failure of Ae. hendersoni to orally transmit La Crosse virus. However, eight parenterally infected Ae. hendersoni females transovarially transmitted the virus to 25% (5 of 20) of their progeny.     

Experimental Venezuelan Equine Encephalomyelitis Virus Infection of the Bovine

Two groups of four dairy cows (Bos taurus) were infected subcutaneously with the epizootic Venezuelan equine encephalomyelitis virus (VEE) strains MF-8 and San Pelayo, respectively. Animals experienced no clinical illness, but all developed significant neutropenia. Virus was recovered once each from the blood of three animals but did not exceed 10(2.2) SMICLD(50) (Suckling mouse intracerebral lethal dose(50))/ml. Specific neutralizing antibodies appeared in the serum of all animals, but there were no significant differences in titers against different naturally occurring VEE subtypes. Dairy cattle thus appear to play no role in virus transmission during VEE epizootics but may serve as retrospective immunological sentinels of virus activity.     

Transmission of Ockelbo virus by Aedes cinereus, Ae. communis, and Ae. excrucians (Diptera: Culicidae) collected in an enzootic area in central Sweden

Studies were conducted to determine the ability of Aedes excrucians (Walker), Ae. cinereus Meigen, and Ae. communis (De Geer) group mosquitoes, collected in an Ockelbo (OCK) virus enzootic area in central Sweden, to transmit this virus. All three species were highly susceptible to infection; at least 96% of the specimens of each species became infected after ingesting blood from a viremic chicken. Recovery of virus from the legs of all 61 of the Ae. excrucians and from 51% (24 of 47) and 75% (6 of 8) of the Ae. cinereus and Ae. communis tested, respectively, indicated that OCK virus readily disseminated from the midgut to the hemocoel in these species. Although none of the Ae. communis refed, Ae. cinereus and Ae. excrucians successfully transmitted OCK virus by bite. Because these Aedes species are attracted to avian and mammalian hosts and because OCK virus has been isolated from field-collected specimens in Sweden and the USSR, Aedes mosquitoes should be considered a possible link between human infections and the enzootic cycle involving birds and Culex and Culiseta mosquitoes.