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The Distribution and Development of Eastern Equine Encephalitis Virus in its Enzootic Mosquito Vector, Culiseta Melanura

Thomas W. Scott^*, Stephen W. Hildreth AND Barry J. Beaty

The timing and sequence of eastern equine encephalitis (EEE) virus replication was studied in the organs from a colony strain of orally infected Culiseta melanura. Three methods of virus assay were used: fluorescent antibody (FA) staining of dissected organs; virus titration in cell culture of whole mosquitoes, dissected organs, hemolymph, and egg rafts; and transmission electron microscopy (TEM) of infected hindguts. EEE virus replicated rapidly in Cs. melanura, first in the posterior midgut, after which it disseminated into the hemocoel where hemolymph transported virus to other organs causing a systemic infection that eventually involved all organs examined, except ovarioles. No initial decrease in virus titer of whole mosquitoes or dissected organs was observed when mosquitoes were collected at daily intervals. Muscle tissue contained the greatest amount of specific fluorescence and the largest aggregates of virus that were visible by TEM. Dissemination of virus occurred rapidly, in some mosquitoes after 17 hours of extrinsic incubation (EI). All infected mosquitoes had disseminated infections after 3 days of EI. Maximum amounts of virus were obtained from whole mosquitoes on the 7th day of EI. FA staining of hindguts was determined to be a rapid and reliable method for detection of EEE virus dissemination and replication in Cs. melanura.

Accepted for publication August 18, 1983.

^* Present address: Department of Entomology, University of Maryland, College Park, Maryland 20742.

Present address: Department of Pediatrics, Infectious Disease Unit, University of Rochester Medical Center, 601 Elmwood Avenue, Rochester, New York 14642.

Present address: Department of Microbiology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, Colorado 80523.

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