title: Isolation and propagation of Trypanosoma brucei gambiense from sleeping sickness patients in south Sudan.
abstract: This study aimed at isolating Trypanosoma brucei gambiense 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 Triladyl as the cryomedium. The samples were stored at -150 degrees C in liquid nitrogen vapour in a dry shipper. Eighteen patient stabilates 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. natalensis and Swiss White, C57/BL and BALB/c mice demonstrated that all rodent breeds were susceptible after the second subpassage and developed a parasitaemia >10(6)/ml by Day 5 post infection. The highest parasitaemias were achieved in C57/BL 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.
authors.affiliation: Trypanosomiasis Research Centre (TRC) of KARI, PO Box 362, Kikuyu, Kenya.
authors.forename: Naomi W N
authors.initials: NW
authors.lastname: Maina
authors.forename: Michael
authors.initials: M
authors.lastname: Oberle
authors.forename: Charles
authors.initials: C
authors.lastname: Otieno
authors.forename: Christina
authors.initials: C
authors.lastname: Kunz
authors.forename: Pascal
authors.initials: P
authors.lastname: Maeser
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Ndung'u
authors.forename: Reto
authors.initials: R
authors.lastname: Brun
mesh_terms: D000293:Adolescent; D000328:Adult; D000818:Animals; D002648:Child; D002675:Child, Preschool; D005260:Female; D006801:Humans; D007223:Infant; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D016513:Mice, SCID; D008875:Middle Aged; D051189:Murinae / Q000469:parasitology*; D017422:Sequence Analysis, DNA; D013397:Sudan; D014347:Trypanosoma brucei gambiense / Q000235:genetics / Q000302:isolation & purification*; D014353:Trypanosomiasis, African / Q000097:blood / Q000134:cerebrospinal fluid