ABSTRACT.- Introduction:Bovine mastitis is the most common disease affecting the dairy industry, with staphylococci being considered as one of the most significant and prevalent causes. This study aimed to assess the presence of staphylococcal subclinical mastitis (SCM) in Uruguayan dairy farms and to identify Staphylococcus aureus (SA) and non-aureus staphylococci (NAS) in milking cows. In addition, the antibiotic susceptibility of isolated staphylococci was evaluated. Methodology:We tested 546 apparently healthy milking cows from 11 farms for detecting SCM using the California Mastitis Test (CMT). The cows were not treated with antibiotics. CMT-positive samples were cultured, and colonies compatible with Staphylococcus spp. were further identified through molecular techniques. The susceptibility of the Staphylococcus spp. isolates against thirteen antibiotics was determined using the disk diffusion method. Results:Subclinical staphylococcal mastitis was present in almost all (82%) farms. SA (n = 39) was more common than NAS (n = 9) in the 48 samples tested. Isolates exhibited resistance to one, two, and even three different antibiotics. Resistance to penicillin was the most frequent among SA (23/39) and NAS (4/9). No staphylococci isolates exhibited resistance to cefoxitin, vancomycin, trimethoprim-sulfamethoxazole, erythromycin, or clindamycin. Conclusions:Staphylococcal SCM is one of the most common diseases in Uruguayan dairy farms. SA was the prevalent pathogen, however SA and NAS mastitis coexisted in many farms. NAS were identified and its distribution was similar to other countries. Penicillin had the highest and most frequent percentage of resistance. Copyright © 2022 de los Santos et al.