The pathogenicity of blood stream and central nervous system forms of Trypanosoma brucei rhodesiense trypanosomes in laboratory mice: a comparative study.

Background: Human African trypanosomiasis (HAT) develops in two stages namely early stage when trypanosomes are found in the blood and late stage when trypanosomes are found in the central nervous system (CNS). The two environments are different with CNS environment reported as being hostile to the trypanosomes than the blood environment. The clinical symptoms manifested by the disease in the two environments are different. Information on whether blood stream are pathologically different from CNS trypanosomes is lacking. This study undertook to compare the inter-isolate pathological differences caused by bloodstream forms (BSF) and central nervous system (CNS) of five Trypanosoma brucei rhodesiense ( Tbr) isolates in Swiss white mice. Methods: Donor mice infected with each of the five isolates were euthanized at 21 days post infection (DPI) for recovery of BSF trypanosomes in heart blood and CNS trypanosomes in brain supernatants. Groups of Swiss white mice (n = 10) were then infected with BSF or CNS forms of each isolate and monitored for parasitaemia, packed cell volume (PCV), body weight, survivorship, trypanosome length, gross and histopathology characteristics. Results: Amplification of SRA gene prior to trypanosome morphology and pathogenicity studies confirmed all isolates as T. b. rhodesiense. At 21 DPI, CNS trypanosomes were predominantly long slender (LS) while BSF were a mixture of short stumpy and intermediate forms. The density of BSF trypanosomes was on average 2-3 log-scales greater than that of CNS trypanosomes with isolate KETRI 2656 having the highest CNS trypanosome density. Conclusions: The pathogenicity study revealed clear differences in the virulence/pathogenicity of the five (5) isolates but no distinct and consistent differences between CNS and BSF forms of the same isolate. We also identified KETRI 2656 as a suitable isolate for acute menigo- encephalitic studies.

Microbiome frequency and their association with trypanosome infection in male Glossina morsitans centralis of Western Zambia.

Tsetse flies (Diptera: Glossinidae) are considered primary cyclical vectors that transmit pathogenic trypanosomes in Africa. They harbour a variety of microbes including Wolbachia, Sodalis and the salivary gland hypertrophy virus (SGHV) which are all vertically transmitted. Knowledge on tsetse microbiome and their interactions may identify novel strategies for tsetse fly and trypanosomiasis control. Area-wide application of such strategies requires an understanding of the natural microbiome frequency in the different species and subspecies of Glossina in their geographical populations. Consequently, this study determined the prevalence of Sodalis, Wolbachia, SGHV and trypanosome infections in Glossina morsitanscentralis from two sites of Western Zambia. We also explored possible associations of the microbes with trypanosome infections. Male G. morsitanscentralis samples were collected from two sites (Lyoni and Lusinina) in Western Zambia. The age structure of the flies at each site was determined using the wing fray method. DNA was extracted from the samples and analyzed for Wolbachia, Sodalis, SGHV and trypanosome presence using PCR. Associations and measures of associations between trypanosome infection and microbes in the fly were determined. The flies from the two locations (Lusinina, n=45 and Lyoni, n=24) had a similar age structure with their median fray category not being significantly different (p=0.698). The overall prevalence of Wolbachia was 72.5% (95% CI: 61.6-83.3%), Sodalis was 15.9% (95% CI: 7.1-24.8%), SGHV was 31.9% (95% CI: 20.6-43.2%) and Trypanosoma species was 23.2% (95% CI: 13-33.4%). The prevalence of Wolbachia was significantly higher in Lusinina than Lyoni (p=0.000). However this was not the case for Sodalis, SGHV and Trypanosoma species. Despite the low number of flies that were positive for both trypanosome and Sodalis (6; 8.7%), a statistically significant association (p=0.013; AOR 6.2; 95% CI: 1.5-25.8) was observed in G. morsitanscentralis. The study showed that the prevalence of microbiota may vary within the same species of the tsetse depending on the geographical location as was the case of Wolbachia. Further it showed that infection with Sodalis could affect vector competence. The study concludes that Sodalis could be an ideal candidate for symbiont-mediated trypanosomiasis control interventions in G. morsitanscentralis.