The use of molecular tools for Pan-Trypanosoma analysis and epigenetics of the host
Ideozu, EJ 2015, The use of molecular tools for Pan-Trypanosoma analysis and epigenetics of the host , PhD thesis, University of Salford Manchester.
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Trypanosomes are a major cause of disease and death in both animal and human populations. Resistance or susceptibility to African trypanosomes has been associated with TLR9 gene. Hence, the primary objectives of this study were to apply molecular tools to investigate trypanosome infections in British badgers and Nigerian cattle, and to investigate variation in bovine TLR9 gene in relation to their trypanosome infection status. The ITS Nested PCR method was used to detect trypanosomes in the British badgers and Nigerian cattle. Two novel hemi-nested PCRs targeting two bovine TLR9 CpG Islands were developed to derive genetic and epigenetic DNA sequence data of 72 African bovine samples. Twenty-nine out of 82 badger samples amplified tested positive for trypanosomes (35.4% prevalence). Analysis of sequence data showed the badgers were infected with Trypanosoma (Megatrypanum) pestanai and as expected our phylogenetic analysis shows the badger trypanosome to cluster together with T. pestanai (100% bootstrap support) in the Megatrypanum clade. Ten out of 80 Southern Nigerian cattle were shown to be positive for trypanosomes resulting in a 12.5% prevalence rate. A total of 9 polymorphisms were found in targeted region of bovine TLR9 gene. The study showed no significant association between SNPs and trypanosomiasis (p = >0.05). Our results confirm the methylation of bovine TLR9 gene and identified CpG-SNPs (2256 and 2865) which removes a C-G site and perhaps could alter DNA methylation as potential epigenetic markers for bovine TLR9 gene. Also, it reports the significant correlation between CpG Island SNPs (p = <0.05, all cases), suggesting possession of one Island SNP is a predicting tool for possession of the others. Future work is targeted at publishing papers in peer reviewed journals based on results from these studies.
|Item Type:||Thesis (PhD)|
|Themes:||Built and Human Environment|
|Schools:||Schools > School of Environment and Life Sciences > Biomedical Research Centre|
|Funders:||Non funded research|
|Depositing User:||EJ Ideozu|
|Date Deposited:||23 Jul 2015 12:03|
|Last Modified:||01 Jul 2016 01:38|
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