Unraveling the genetic diversity of creole breeds is central to understanding their value and importance for conservation and improvement in diverse production environments in America. In most of its countries, information on attributes of many livestock breeds is scanty and do not support well-informed decision-making upon relevant management strategies. This study aimed at investigating the “Paisanita” cattle breed, a landrace breed adapted to different ecological regions of Guatemala. For this purpose, a total of 47 adult females belonging to 4 different geographical areas of the country were analysed on the basis of 24 quantitative traits. Results showed no differences between areas and so, the possible presence of ecotypes was discarded. This conclusion helps the objectives of conservation, and future monitoring of “Paisanita” cattle.