ABSTRACT Páramo ecosystems are of great importance because they are considered hotspots within the Tropical Andes. They are also very important for their role as producers and regulators of water processes in the Neotropic. However, the human occupation of the Colombian Páramos has generated conflict between environmental benefits and productive land uses, specifically the potato cultivation and livestock. To assess possible changes associated with potato cultivation (Solanum tuberosum L.) and livestock on the microbial communities of Páramo soils, the objective of this research was to evaluate the possible effects of potato cultivation and livestock farming on the soil microorganisms associated with different functional groups (nitrogen fixers, phosphate solubilizers and cellulolytic) in the Páramo of Nevados National Natural Park (Nevados NNP), Colombia. Samples were collected from soils under potato cultivation, livestock, and Páramo conservation areas over two climatic seasons (rainy and dry) in three farms at different elevations (3769, 3590, and 3432 m a.s.l.). The microorganisms were isolated using selective culture media for each functional group and identified using molecular markers; microbial diversity was analyzed using multivariate statistical tools. Changes were dependent on land use, elevation, and climate and were statistically significant in the rainy season on all three farms and one of the farms during the dry season. Similarly, the results indicated that climate has a greater impact on the evaluated microbial communities than land use does; the changes were significantly different between the soil under potato cultivation and in conserved Páramo sites at most of the evaluated locations and between soil subjected to livestock farming and Páramo in certain locations. However, the differences between potato cultivation and livestock farming were smaller. This study showed for the first time that the microbial structure (abundance and composition) of microorganism functional groups was different as a result of potato cultivation and livestock farming on Páramo soils, although these changes were dependent on farm elevation and climate.