Since Þrst discovered in the New York City area in 1999, West Nile virus (WNV) has become established over much of the continental United States and has been responsible for Ͼ10,000 cases of severe disease and 400 human fatalities, as well as thousands of fatal infections in horses. To develop appropriate surveillance and control strategies, the identiÞcation of which mosquito species are competent vectors and how various factors inßuence their ability to transmit this virus must be determined. Therefore, we evaluated numerous mosquito species for their ability to transmit WNV under laboratory conditions. This report contains data for several mosquito species not reported previously, as well as a summary of transmission data compiled from previously reported studies. Mosquitoes were allowed to feed on chickens infected with WNV isolated from a crow that died during the 1999 outbreak in New York City. These mosquitoes were tested Ϸ2 wk later to determine infection, dissemination, and transmission rates. All Culex species tested were competent vectors in the laboratory and varied from highly efÞcient vectors (e.g., Culex tarsalis Coquillett) to moderately efÞcient ones (e.g., Culex nigripalpus Theobald). Nearly all of the Culex species tested could serve as efÞcient enzootic or amplifying vectors for WNV. Several container-breeding Aedes and Ochlerotatus species were highly efÞcient vectors under laboratory conditions, but because of their feeding preferences, would probably not be involved in the maintenance of WNV in nature. However, they would be potential bridge vectors between the avianÐCulex cycle and mammalian hosts. In contrast, most of the surface pool-breeding Aedes and Ochlerotatus species tested were relatively inefÞcient vectors under laboratory conditions and would probably not play a signiÞcant role in transmitting WNV in nature. In determining the potential for a mosquito species to become involved in transmitting WNV, it is necessary to consider not only its laboratory vector competence but also its abundance, host-feeding preference, involvement with other viruses with similar transmission cycles, and whether WNV has been isolated from this species under natural conditions.

Culex salinarius is a bridge vector to humans, while Cx. pipiens and Cx. restuans are more efficient enzootic vectors.

n Ixodes scapularis is the principal vector of Lyme disease on the East Coast and in the upper Midwest regions of the UnitedStates, yet the tick is also present in the Southeast, where Lyme disease is absent or rare. A closely related species, I. affinis, also carries the pathogen in the South but does not seem to transmit it to humans. In order to better understand the geographic diversity of the tick, we analyzed the microbiota of 104 adult I. scapularis and 13 adult I. affinis ticks captured in 19 locations in South Carolina, North Carolina, Virginia, Connecticut, and New York. Initially, ticks from 4 sites were analyzed by 454 pyrosequencing. Subsequently, ticks from these sites plus 15 others were analyzed by sequencing with an Illumina MiSeq machine. By both analyses, the microbiomes of female ticks were significantly less diverse than those of male ticks. The dissimilarity between tick microbiomes increased with distance between sites, and the state in which a tick was collected could be inferred from its microbiota. The genus Rickettsia was prominent in all locations. Borrelia was also present in most locations and was present at especially high levels in one site in western Virginia. In contrast, members of the family Enterobacteriaceae were very common in North Carolina I. scapularis ticks but uncommon in I. scapularis ticks from other sites and in North Carolina I. affinis ticks. These data suggest substantial variations in the Ixodes microbiota in association with geography, species, and sex.

Two hundred and ten isolations of West Nile virus (WNV) were obtained from 17 mosquito species in six genera in statewide surveillance conducted in Connecticut from June through October, 1999-2003. Culex pipiens (86), Culex salinarius (32), Culex restuans (26), Culiseta melanura (32), and Aedes vexans (12) were implicated as the most likely vectors of WNV in the region based on virus isolation data. Culex pipiens was abundant from July through September and is likely involved in early season enzootic transmission and late season epizootic amplification of the virus in wild bird populations. Epidemic transmission of WNV to humans in urban locales is probable. The abundance of Cx. restuans in June and July and isolations of WNV in early July suggest that this species may play an important role as an enzootic vector involved in early amplification of WNV virus among wild birds. Its involvement as a bridge vector to humans is unlikely. Culex salinarius was the most frequently captured Culex species and was abundant in August and September when virus activity was at its height. Frequent isolations of WNV from this species in September when the majority of human cases were reported in union with its abundance at this time of the year, demonstrated vector competence, and broad feeding habits, make Cx. salinarius a likely bridge vector to humans, horses and other mammals. Multiple isolations WNV from Cs. melanura collected in more rural locales in late August and September, provide supportive evidence to suggest that this predominant avian feeder may play a significant role in epizootic amplification of the virus among wild bird populations in these environs. Aedes vexans was the only species of Aedes or Ochlerotatus from which multiple isolations of WNV were made in more than one year and was among the most frequently trapped and abundant species throughout the season. Since Ae. vexans predominately feeds on mammals it is unlikely to play a significant role in epizootic amplification of WNV, however, because of its abundance and aggressive mammalian and human biting behavior it must receive strong consideration as a bridge vector to humans and horses. The occasional virus isolations obtained from Aedes cinereus (4), Uranotaenia sapphirina (3), Ochlerotatus canadensis (2), Ochlerotatus trivittatus (2), Ochlerotatus sollicitans (2), Ochlerotatus sticticus (2), Psorophora ferox (2), Anopheles punctipennis, Anopheles walkeri, Ochlerotatus cantator, Ochlerotatus taeniorhynchus, and Ochlerotatus triseriatus in conjunction with their inefficient vector competency and host feeding preferences indicate that these species likely play a very minor role in either the enzootic maintenance or epizootic transmission of WNV in this region. The principal foci of WNV activity in Connecticut were identified as densely populated (>3,000 people/mi2) residential communities in coastal Fairfield and New Haven Counties, and in the case of 2002, similar locales in proximity of the city of Hartford in central Hartford County. In almost all insta...

West Nile (WN) virus, a mosquito-transmitted virus native to Africa, Asia, and Europe, was isolated from two species of mosquitoes, Culex pipiens and Aedes vexans, and from brain tissues of 28 American crows, Corvus brachyrhynchos, and one Cooper's hawk, Accipiter cooperii, in Connecticut. A portion of the genome of virus isolates from four different hosts was sequenced and analyzed by comparative phylogenetic analysis. Our isolates from Connecticut were similar to one another and most closely related to two WN isolates from Romania (2.8 and 3.6 percent difference). If established in North America, WN virus will likely have severe effects on human health and on the health of populations of birds.