Identification of Trypanosoma congolense genes with metacyclic stage-specific expression through transcriptomic analysis of wild-caught tsetse flies (Glossina palpipes).


K Mathenge1; A P Jackson2
1 Institute of Infection and Global Health, UK;  2 University of Liverpool, UK


Animal African trypanosomiasis is a lethal disease of wild and domestic animals, endemic throughout sub-Saharan Africa and caused by African trypanosomes (Trypanosoma spp.). In Eastern Kenya, T. congolense is transmitted by tsetse flies (G. palpipes), when infectious metacyclic-stage parasites that reside within the fly mouthparts are introduced while feeding. To better understand the developmental regulation of parasite genes during the infectious metacyclic stage, we used RNA-seq to estimate transcript abundance in dissected mouthparts, and then DE-seq to identify transcripts preferentially expressed in metacyclic cells. Tsetse flies were caught using baited traps in the Shimba Hills national park, Kenya, and metacyclic trypanosomes were sluiced from dissected mouthparts. Extracted RNA was sequenced on the Illumina Hiseq4000 platform and mapped to the T. congolenseIL3000 PacBio reference genome. Our analysis has identified 793 genes, transcripts of which are significantly over-represented in metacyclic-stage cells. These include diverseT. congolense-specific genes, such as a Variant Surface Glycoprotein (VSG) sequence, that might offer new opportunities for vaccine development, as well as new insights into the molecular host-parasite interactions during the early stages of trypanosome infection following tsetse bite. 

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