We investigated the distribution and antimicrobial resistance of 120 Staphylococcus felis isolates from feline patients in Japan, mainly from the urinary tract (28.3%), abscesses (23.3%), ears (22.5%), and nasal cavity (10.8%). The distribution of S. felis differed from those of previous studies in Japan and other countries. Antimicrobial susceptibility testing revealed a relatively high resistance to penicillin (PEN, 33.3%), followed by erythromycin (ERY, 15.8%), clindamycin (CLI, 13.3%), and levofloxacin (5.0%). However, oxacillin resistance was not detected. Notably, 11/120 (9.2%)S. felis isolates exhibited multidrug resistance, i.e., resistance to more than three classes of drugs, which mainly consisted of PEN-ERY-CLI resistance phenotypes. This is the first investigation on antimicrobial-resistant S. felis isolates from feline patients in Japan.