In vivo and in vitro sensitivity of Trypanosoma evansi and T. equiperdum to diminazene, suramin, MelCy, quinapyramine and isometamidium

Abstract

The sensitivity of three Trypanosoma equiperdum clones and thirteen Trypanosoma evansi clones originating from the People's Republic of China, the Philippines, Ethiopia and elsewhere to a series of drugs was determined in vivo and in vitro. The drugs tested were diminazene aceturate (Berenil), suramin (Naganol), MelCy (Cymelarsan), quinapyramine sulfate (Trypacide) and isometamidium chloride (Samorin). The activity of each drug was expressed as: 1) in vitro: the minimal effective concentration which killed trypanosome population by 100% within 24 h of drug exposure (MEC100); the maximum tolerated concentration in which trypanosomes could propagate at the same rate as the controls during 48 h of drug exposure (MTC100); 2) in vivo: the curative dosage in 100% of infected mice (CD100); the highest ineffective dosage: 100% of infected mice remain infected (ID100). MEC100 values of diminazene aceturate ranged from 0.0556 microgram/ml to 14.24 micrograms/ml for the eleven tested clones (differed by 256-fold); CD100 values of this drug ranged from 2.25 mg/kg to greater than 89 mg/kg (differed by greater than 40-fold). Diminazene aceturate at up to 89 mg/kg had no effect on T. evansi SHBR, T. equiperdum PBR (Berenil resistant organisms selected by continual passage of the organisms through mice treated with increasing concentrations of drug), or T. evansi AH (strain isolated in the field). Comparable MEC100 values for other trypanocides tested were 1-8 micrograms/ml for suramin, 0.005-0.04 microgram/ml for MelCy, 1-16 micrograms/ml for quinapyramine sulfate and 1-4 micrograms/ml for isometamidium chloride. Clones selected for resistance to diminazene aceturate were not cross-resistant to suramin and isometamidium chloride. In contrast, the clones resistant to diminazene were shown to be more sensitive to quinapyramine sulfate than the normal strains in in vivo tests. The results indicate that resistance to diminazene aceturate by T. evansi and T. equiperdum clones in vivo also occurred in vitro. Resistance to isometamidium chloride in the clones tested in vivo was not observed in vitro, except for T. equiperdum SA. It therefore appears that drug bioavailability is altered or drug biotransformation occurs during the in vivo test. We conclude that the in vitro assay procedure may be of potential use for screening new trypanocides and also for the rapid detection of drug resistant isolates of T. evansi and T. equiperdum.