A drug incubation infectivity test (DIIT) for assessing resistance in trypanosomes

Blood stream forms of drug-resistant and sensitive Trypanosoma brucei brucei, Trypanosoma brucei evansi and Trypanosoma vivax were incubated in a liquid medium for 24 h at 37 degrees C in the presence of various concentrations of diminazene aceturate (Berenil) or isometamidium chloride (Samorin), and assayed for infectivity in mice. Whereas the infectivity to mice of all Samorin-sensitive trypanosomes was decreased after incubation with 1 ng Samorin ml-1, the Samorin-resistant stocks remained infective for mice. Two of the Samorin-resistant stocks remained infective after incubation with Samorin concentrations of up to 50 ng ml-1. The infectivity of Berenil-resistant trypanosome stocks were also retained after incubation with drug concentrations (0.5 or 1.0 micrograms ml-1) which otherwise inhibited the infectivity of Berenil-sensitive trypanosome stocks. In addition, differences in infectivity were observed when Berenil-resistant and sensitive trypanosome stocks were incubated in medium supplemented with serum from goats previously treated with Berenil. Thus, drug-resistant and sensitive trypanosomes can be clearly distinguished using the drug incubation infectivity test.