Abstract/Summary

Arenaviruses are enveloped RNA viruses with a genome of two segments of single stranded RNA which is classified as ambisense in its coding strategy. Arenaviruses within the Reptarenavirus genus infect many species of snake and are associated with inclusion body disease (IBD). Reptarenavirus infection of humans has not been reported. In this thesis molecular techniques were utilized for RNA analysis and gene amplification by reverse transcription PCR (RT-PCR) in order to find evidence of reptarenavirus sequences in reptilian samples. Total RNAs were isolated from frozen specimens of snakes and other reptiles followed by an evaluation of RNA integrity. Degenerate primers of ribosomal mitochondrial and reptarenavirus genes were used for quality control of the RNA and for identification of virus sequence respectively, and then the results were confirmed by Sanger sequencing. The mitochondrial RNA was successfully amplified, whereas no reptarenavirus sequences were found in the samples screened. Spiking experiments determined that the cut-off for detection was less than 108 copies of virus RNA per sample. Although no evidence for the circulation of reptarenaviruses was found the study serves as a guide for reptarenavirus investigations using the RT-PCR technique in cases where new samples are suspected of having been infected with an existing or novel reptarenavirus. In addition, a related study investigating the role of replication organelles in the pathogenicity of infectious bronchitis virus (IBV) was performed using Transmission Electron Microscopy (TEM) of infected cells. Further, an assessment was made of the use of the arenavirus L sequence to look for related sequences associated with currently unknown viruses in the TSA database. Several matches were found which, along with a reasoned approach to their phyla, was used to provide a likelihood score of their zoonotic potential.