Populations from different countries may present different cellular modifications among themselves, and the Buccal Micronucleus Cytome (BMCyt) assay in human buccal mucosal tissue may be a marker to evaluate these modifications. This study evaluated and compared DNA damage and other nuclear anomalies between Brazilian and Spanish gym users. This is a cross-sectional study carried out with gym users of Santa Cruz do Sul/Brazil and Madrid/Spain. The BMCyt assay was performed for biomarkers of DNA damage (micronuclei and/or nuclear buds), cytokinetic defects (binucleated cells), proliferative potential (basal cell frequency) and/or cell death (condensed chromatin, karyorrhexis, pyknotic and karyolytic cells) in human buccal mucosal. Of the 228 individuals evaluated, 163 were Brazilian, and 65 were Spanish. Gym users of both countries differed between weight, body mass index, body fat, and muscle mass. The Brazilians presented a significantly higher frequency of micronuclei, nuclear buds, cells with condensed chromatin and karyorrhexis. Spaniards, however presented a significantly higher frequency of karyolytic cells. In conclusion, Brazilian gym users presented significantly higher rates of DNA damage and cell death, while the Spanish presented a higher frequency of advanced stage cell death.