Year

2013

Season

Spring

Paper Type

Master's Thesis

College

College of Arts and Sciences

Degree Name

Master of Science in Biology (MS)

Department

Biology

NACO controlled Corporate Body

University of North Florida. Department of Biology

First Advisor

Dr. Doria F. Bowers

Second Advisor

Dr. James J. Gelsleichter

Third Advisor

Dr. Raquel Hernandez

Department Chair

Dr. Daniel C. Moon

College Dean

Dr. Barbara A. Hetrick

Abstract

In the present investigation two container breeding mosquitoes, Aedes aegypti and Aedes albopictus were exposed to Sindbis virus (SINV). Mosquitoes were proffered a viremic bloodmeal using a vertical membrane feeder in an attempt to infect females while preserving virus-gut interactions that are speculated to be responsible for amplification and selective roles encountered in nature. Cohorts of 50 mosquitoes of each species were exposed to SINV strain TR339 or EMEM in a bloodmeal and incubated in insectary conditions for 30 days. Once infected, the mosquito remains persistently infected for life, accumulating mutations in the virus RNA genome. Plaque size variants may indicate changes to the virus genome, therefore in this investigation TR339 persistent disseminated isolates (TR339-PDIs) were isolated from mosquito legs 30 days post infection (p.i.) and assayed for plaque size differences. These TR339-PDIs represent virus selected for by the whole mosquito. TR339-PDIs following a long term infection were used to study virus adaptations selected for during persistent infection of the mosquito host. The two mosquito species exhibited different host responses to SINV; survivorship, dissemination rates, and immunohistochemistry of mosquito midguts and hindguts were assayed at 30 days p.i. Aedes aegypti and Ae. albopictus along with SINV strain TR339 provide a good model system to study the selective pressures that occur within the whole mosquito. Greater knowledge of the mechanisms of virus persistence in the mosquito host is vital for controlling arthropod-borne disease at the vector level.

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