A thesis submitted to the Faculty of Science, University of the Witwatersrand, Johannesburg, in fulfillment of the requirements for the degree of Doctor of Philosophy. / The aim of this thesis was to develop DNA probes to distinguish between the protozoan parasites Theileria parva parva and T. p. lawrencei which cause East Coast fever (ECF) and Corridor disease respectively. ECF was eradicated from South Arrlca in 1954, and today Corridor disease has become the most important form of theileriosis. Although ECF has been eradicated, the vector ticks are still prevalent in South Africa and the cattle population would be highly susceptible to a recurrence of the disease, At present there is no reliable means of distinguishing between T.p. parva and T. p. lawrencei. Sequence differences between T. parva and other Theileria species have previously been found in the small subunit ribosomal RNA (rRNA) gene; probes designed to detect these sequence differences Can be used to distinguish between Theileria species. We therefore decided to search for differences in the rRNA genes of T. p. parva and T.p. lawrencei. To this end, the entire "RNA transcription unit was amplified from a cloned T. p, lawrence; parasite; the unit comprises the small subunit rRNA (SSUrRNA) gene, the internal transcribed spacer (ITS) and the large subunit rRNA (LSUrRNA) gene. The amplification products were cloned and sequenced, and the T.p, lawrencei rRNA sequence was compared to that of T. p, parva, While there was little variation in their SSUrRNA and LSUrRNA gene sequences, there was major sequence variation in the ITS The ITSs from twelve T. parva isolates were amplified, cloned and sequenced, and eleven characterisation oligonucleotide probes were identified. The T. p, parva isolates screened in this study hybridised with a limited subset of the probes, While the T. p. lawrencei isolates, hybridised with many more of the probes, indicating that the T. parva population in cattle is more homogenous than that in buffalo. There thus appears to have been a selection in cattle of a relatively homogenous subpopuiation of T. parva from a much larger, more diverse gene pool in buffalo. Although most T.p. parva isolates (93.5%) were detected by probe TPPI, and most T.p, lawrencei isolates (81.8%) were detected by / AC2017

Text in English / Corridor disease is an economically important tick-borne disease of cattle in southern Africa. The disease is caused by Theileria parva and transmitted by the vectors, Rhipicephalus appendiculatus and R. zambeziensis. There is currently no vaccine to protect cattle against T. parva that is permitted in South Africa. To develop recombinant anti-tick vaccines against Corridor disease, comprehensive databases of genes expressed in the tick’s salivary glands are required. Therefore, in Chapters 2 and 3, mRNA from the salivary glands of R. appendiculatus and R. zambeziensis was sequenced and assembled using next generation sequencing technologies. Respectively, 12 761 and 13 584 non-redundant protein sequences were predicted from the sialotranscriptomes of R. appendiculatus and R. zambeziensis and uploaded to public sequence domains. This greatly expanded the number of sequences available for the two vectors, which will be invaluable resources for the selection of vaccine candidates in future. Further, in Chapter 3, differential gene expression analysis in R. zambeziensis revealed dynamic expression of secretory protein transcripts during feeding, suggestive of stringent transcriptional regulation of these proteins. Knowledge of these intricate expression profiles will further assist vaccine development in future. In Chapter 4, comparative sialotranscriptomic analyses were performed between R. appendiculatus and R. zambeziensis. The ticks have previously shown varying vector competence for T. parva and this chapter presents the search for correlates of this variance. Phylogenetic analyses were performed using these and other publically available tick transcriptomes, which indicated that R. appendiculatus and R. zambeziensis are closely related but distinct species. However, significant expression differences were observed between the two ticks, specifically of genes involved in tick immunity or pathogen transmission, signifying potential bioinformatic signatures of vector competence. Furthermore, nearly four thousand putative long non-coding RNAs (lncRNAs) were predicted in each of the two ticks. A large number of these showed differential expression and suggested a potential transcriptional regulatory function of lncRNA in tick blood feeding. LncRNAs are completely unexplored in ticks. Finally, in Chapter 5, concluding remarks are given on the potential impact the R. appendiculatus and R. zambeziensis sialotranscriptomes may have on future vaccine developments and some future research endeavours are discussed. / Life and Consumer Sciences / Ph. D. (Life Sciences)