Mosquito surveillance is an integral part of understanding mosquito-borne disease, including and not limited to the La Crosse virus. The primary container-inhabiting Aedeni vectors include Aedes triseritatus, Aedes albopictus, and Aedes japonicus. To better understand the efficacy of gravid traps and oviposition traps as surveillance methods for these mosquitoes, field studies were conducted in three different habitat types. Ae. triseriatus, Ae. albopictus, Ae. japonicus, and Aedes aegypti eggs were described with the aid of color images and SEM. All eggs were broadly cigar shaped with Ae. triseriatus and Ae. japonicus eggs being dull or matte black while Ae. albopictus and Ae. aegypti eggs were shiny jet black. Ae. triseriatus eggs were larger, lighter in color, and have a rougher appearance when compared to Ae. japonicus. Ae. albopictus and Ae. aegypti can be differentiated by the distinct presence of a micropylar collars in Ae. aegypti. Ovitrap and gravid trap efficacy for the surveillance of Ae. triseriatus, Ae. albopictus, and Ae. japonicus were measured in three different habitats on two different sites. Both sites contained the same 2ha habitats with varying degrees of forest canopy disturbance. Ae. triseriatus was the most abundant mosquito on all sites. Ovitraps and gravid traps were efficient in collecting Ae. triseriatus, while ovitraps were not efficient in collecting Ae. japonicus and Ae. albopictus. / Master of Science in Life Sciences

Public Health and clinical laboratories occasionally obtain viral isolates that cannot be typed by routine methods. Therefore, the sequence-independent, single primer amplification (SISPA) technique was adapted to rapidly identify and characterize viral isolates of unknown etiology. A panel of known (West Nile virus and St. Louis encephalitis virus) and unknown viral isolates (environmental samples collected in Florida) were used to develop and refine the SISPA technique. Selectivity for viral genomic sequences was obtained through enriching viral particles by centrifugation, removal of cellular debris by filtration and removal of host genomic material by benzonase application. The SISPA method successfully amplified the panel of known viruses and a previously unknown environmental viral isolate. The previously unknown environmental viral isolate was determined to be closely related, if not identical, to Flanders virus, a member of Rhabdoviradae. A Flanders virus specific RT-PCR assay identified a total of five previously unknown environmental viral isolates as Flanders virus. Unidentified viral isolates were obtained during arbovirus surveillance efforts in Florida, either from the Florida Department of Health program (BOL-Tampa) during 2005 – 2009, or collected during an ongoing project at the University of South Florida studying the ecology of arthropod-borne encephalitis viruses at sites located in Florida. In a concurrent study, SISPA was successfully used to characterize an unidentifiable virus isolate related to members of the Bunyaviradae family which was designated as Infirmatus virus. Natural mosquito population (10,557 mosquitoes) collected in Florida was screened for Flanders virus and members of Bunyaviradae to determine infection prevalence. Although Flanders virus was not detected in this population, Infirmatus virus was identified in 14 mosquito pools with the highest infection prevalence in Cx. quinquefasciatus mosquitoes. The SISPA technique was successful for the genetic identification of unknown viral isolates and application of this method to samples with suspected or unidentified viral etiologies may be used to enhance public health surveillance of emerging or re-emerging viruses in Florida.

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