Host associations of mosquitoes at eastern equine encephalitis virus foci in Connecticut, USA

John J. Shepard, Theodore G. Andreadis, Michael C. Thomas, Goudarz Molaei

Eastern equine encephalitis virus (EEEV) is a highly pathogenic mosquito-borne arbovirus, with active transmission foci in freshwater hardwood swamps in eastern North America, where enzootic transmission is maintained between the ornithophilic mosquito, Culiseta melanura, and wild passerine birds. The role of other locally abundant mosquito species in virus transmission and their associations with vertebrate hosts as sources of blood meals within these foci are largely unknown but are of importance in clarifying the dynamics of enzootic and epidemic/epizootic transmission. Blood-engorged mosquitoes were collected from resting boxes at four established EEEV foci in Connecticut during 2010-2011. Mosquitoes were identified to species, and the identity of vertebrate hosts was determined based on mitochondrial cytochrome b and/or cytochrome c oxidase subunit I gene sequences of polymerase chain reaction products. The vertebrate hosts of 378 (50.3 % of engorged mosquitoes) specimens, representing 12 mosquito species, were identified. Culiseta morsitans (n = 54; 67.5 %), Culex restuans (n = 4; 66.7 %), and Cx. pipiens (n = 2; 100 %) acquired blood meals exclusively from avian hosts, whereas Aedes cinereus (n = 6; 66.7 %), Ae. canadensis (n = 2; 100 %), and Ae. stimulans (n = 1; 100 %) obtained blood meals solely from mammals. Species that fed opportunistically on both avian and mammalian hosts included: Ae. thibaulti (n = 21 avian, and n = 181 mammalian; 57.2 %), Anopheles punctipennis (n = 8 and n = 40; 44.0 %), An. quadrimaculatus (n = 1 and n = 23; 35.7 %), Coquillettidia perturbans (n = 3 and n = 3; 46.2 %) and Ae. abserratus (n = 1 and n = 2; 23.1 %). Culex territans obtained blood meals from avian and amphibian hosts (n = 18 and n = 5; 26.6 %). Mixed blood meals originating from both avian and mammalian hosts were identified in An. quadrimaculatus (n = 1), and Cx. territans (n = 2). Our findings indicate that wood thrush, tufted titmouse, and a few other avian species serve as hosts for mosquitoes, and likely contribute to amplification of EEEV. Our study supports the role of Cs. morsitans in enzootic transmission of EEEV among avian species. Culex territans will seek blood from multiple vertebrate classes, suggesting that this species may contribute to epizootic transmission of the virus. Our findings support roles for Cq. perturbans and An. quadrimaculatus as epidemic/epizootic vectors to humans, horses, and white-tailed deer. Despite its abundance, the potential of Ae. thibaulti to serve as a "bridge vector" for EEEV remains unclear in the absence of any definitive knowledge on its competency for the virus. The contribution of white-tailed deer to the dynamics of EEEV transmission is not fully understood, but findings indicate repeated exposure due to frequent feeding by vector competent mosquito species.