Isolation and propagation of Trypanosoma Bruce gambiense from sleeping sickness patients in south Sudan

Abstract

This study aimed at Isolating Trypanosoma brucei gamblense from human African trypanosomiasis (HAT) patients from south Sudan. Fifty HAT patients Identified during active screening surveys were recruited, most of whom (49/50) were in second-stage disease" Blood and cerebrospinal fluid samples collected from the patients were cryopreserved using Tryadyl as the cryomedium. The samples were stored at -150' C In liquid nitrogen vapour in a dry shipper. Eighteen patient stablates could be propagated in Immunosuppressed Mastomys natalensis and/or SCID mice. Parasitaemia was highest In SCID mice. Further subpassages In M. natalensis increased the virulence of the trypanosomes and all 18 isolates recovered from M. natalensis or SCID mice became infective to other immunosuppressed mouse breeds" A comparison of Immunosuppressed M. nataiensis and Swiss White, C571BL and BALBlc mice demonstrated that all rodent breeds were susceptible after the second subpassage and developed a parasitaemia >106/ml by Day 5 post infection. The highest parasitaemias were achieved in C571BL and BALB/c mice. These results indicate that propagation of T. b" gambiense isolates after Initial isolation In Immunosuppressed M. natalensis or SCID mice can be done in a range of Immunosuppressed rodents.