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Polymorphism of Attachin D Gene in Glossina pallidipes Tsetse Fly Populations of Nguruman and Busia-Teso, Kenya

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dc.contributor.author Mayoke, Abraham
dc.date.issued 2011-03
dc.date.accessioned 2022-09-28T12:20:21Z
dc.date.available 2022-09-28T12:20:21Z
dc.description.abstract Tsetse flies (Glassina spp) that spread African trypanosomes express Attacin, an immune system protein, during trypanosome infection. The actual mechanism of protection by Attacin proteins in trypanosome-infected Glossina pallidipes is unknown. The objectives for this study was to assess polymorphism of Attacin gene in natural G. pallidzpes populations isolated from Nguruman and Busia-Teso regions of Kenya (regions with different disease transmission levels and genetic diversity of tsetse flies) and correlate polymorphism of the gene to trypanosome infection in G. pallidzpes from the study regions. Biconical traps and suitable attractants were used to lure and capture the G. pallidipes flies. Midguts were dissected from the captured flies and preserved in Trizol reagent. Trypanosome parasitemia in the salivary glands of the flies were determined through standard microscopy. PCR of Attacin D (AttD) gene was conducted on Genomic DNA (extracted from carcasses), and the product sequenced using automated sequencer. Trypanosome infections in the rnidgut tissues were detected by microscopy and their presence confirmed via nested RT-PCR using trypanosome species specific primers. A total of 586 non-teneral and l5l teneral G. pallidipes were sampled fiom Nguruman. In Busia ~ Teso, 17 and 16 non-teneral and teneral G. palidipes were sampled. A homologue of G. m. morsitans Attacin D in G. pallidzpes was successfully identified fiom a region of 484 base pairs. Most polymorphism were mutations, deletions and substitutions but with predominance of mutation in coding sites. Two types of AttD were revealed, a conserved region and several introns in all individual flies’ sequences in position 170 to 180. Attacin D polymorphism in G. pallidzpes from Nguruman and Busia-Teso were predominant downstream from position 199 to position 298. Microcopy examination revealed putative presence of trypanosome in midgut and proboscis of two flies, one each from Nguruman and Busia~Teso, while PCR results revealed putative infections in two and four flies from respective towns. Putative G. pallidipes Attacin D homologous were identified only in G. m. morsitans and in Aedes aegypti among insect vectors of pathogens in Glossina, Aedes and Rhodnius genera. Interproscan analyses did not reveal any differences in domain architecture among the putative G. pallidipes attacin D sequences. The observed differences in polymorphism of Atttacin D populations in Ngururnan and Busia-Teso may be responsible for the differences in Trypanosomiasis incidences in the two towns. en_US
dc.description.sponsorship Biosciences Eastern and Central Africa through Afiica Union-NEPAD and Canadian International Development Agency en_US
dc.language.iso en en_US
dc.publisher Egerton University en_US
dc.subject Polymorphism -- Attachin D Gene -- Tsetse Fly en_US
dc.title Polymorphism of Attachin D Gene in Glossina pallidipes Tsetse Fly Populations of Nguruman and Busia-Teso, Kenya en_US
dc.type Thesis en_US

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