The aim of this article is to provide an overview of the antimicrobial resistance of S. uberis, which is today one of the most important pathogens causing mammary gland inflammations in the Czech dairy cows. In the course of 2017 and 2018, 3,719 quarter milk samples from clinical and subclinical mastitis from 112 farms in the Czech Republic were examined. S. uberis was the most commonly isolated mam­mary gland pathogen, isolated in 17.4% of the samples, making up 28.4% of gram-positive isolates. The susceptibility to ten antimicrobials was evaluated in 163 S. uberis isolates by measuring their minimum inhibitory concentration. Isolates of S. uberis showed resistance mainly to tetracycline (63.2%), strepto­mycin (52.1 %), clindamycin (30.1 %) and rifampicin (2.5%).The high percentage of intermediate suscep­tible isolates were reported for rifampicin (63.2%) and penicillin (35%). All S. uberis strains were suscep­tible to gentamicin and trimethoprim-sulfamethoxazole. However, only 6.7% of the isolates were sus­ceptible to all antimicrobials tested and 38.7% of the isolates were multidrug resistant. The results of this work provide a comprehensive overview of the susceptibility of S. uberis isolates that occur in Czech farms and are intended to assist veterinarians in deciding on appropriate antibiotic treatment of S. uberis intramammary infections.