Genetic variability in the protozoan causative agent of Chagas' disease, Trypanosoma cruzi, has been analysed in some Latin American countries; Brazil, Bolivia, Chile and Paraguay. Although Colombia is a country displaying enormous biological diversity, few studies have been conducted from the perspective of the population genetics of Trypanosoma cruzi. This study was carried out using 23 Colombian stocks of this protozoan, analysed for 13 isoenzyme loci. The main population genetic results were: (1) Colombia is one of the distribution areas where T. cruzi appears to have the highest genetic variability and heterogeneity in Latin America; (2) the Pgm locus was found in fixed heterozygosis, supporting the presence of diploidy in this organism; (3) the absence of segregating genotypes and the absence of Hardy-Weinberg equilibrium support the view of the existence of a clonal structure as claimed by Tibayrenc and Ayala. Nevertheless, one characteristic of clonal structure, that of over-representation of some identical zymodemes in vast areas of varying environmental conditions, was not found in Colombia. In this country, a strong spatial autocorrelation, with a classic structure of regional patches, was observed.