Catria is one of the 22 native Italian horse breeds that now survive from a larger number. 30 individuals, representative of the Catria horse, were analyzed for 11 microsatellites and compared to data of 10 breeds reared in Italy. Three different approaches, genetic distances, correspondence analysis and clustering methods, were considered to study genetic relationships among Catria and the other horse populations. Genetic differentiation among breeds was highly significant (P < 0.01) for all loci. Average FST values indicate that around 10% of the total genetic variation was explained by the between-breed differences and the three approaches utilized gave similar results. Italian native breeds are clearly separated from the other examined breeds. However, by the correspondence analysis, the Catria appears closer to Maremmano and Murgese. The results of Bayesian approaches give further information showing for Catria a common origin with Maremmano and Italian Heavy Draught. Genetic relationships among Catria and the other breeds are consistent with the breed’s documented history. The data and information found here can be utilized in the organization of conservation programmes planned to reduce inbreeding and to minimize loss of genetic variability.