Trypanosomal infections in female Glossina morsitans morsitans were investigated in an area in the Eastern Province of Zambia between 1992 and 1994. A total of 4416 flies were captured, aged using the ovarian ageing method and screened for trypanosomal infections in both the mouthparts, salivary glands and the midgut. Congolense-type infections were identified in 4.8% of the flies. Vivax-type and immature infections were identified in 1.8% and 6.8% of the flies, respectively. The prevalence of congolense-type, vivax-type and immature infections increased with age. For vivax-type infections the age-prevalence relationship could be described by a model assuming a constant per capita rate of infection. For congolense-type and midgut infections, a polynomial term was added to the model significantly improving the fit. The per capita at which flies become infected was significantly higher for immature compared to mature infections. Observations strongly suggest that tsetse acquire new midgut infections at any age and that maturation of these infections is not limited to those obtained during the first blood meal.