To describe the antimicrobial susceptibility patterns of the most commonly isolated bacteria cultured from synovial fluid samples from horses with suspected septic synovitis treated at an equine referral hospital between May 1, 2008, and September 24, 2017. 131 synovial fluid samples from 108 client-owned horses. A retrospective medical record search was conducted to identify horses with suspected septic synovitis and results of synovial fluid bacterial culture and antimicrobial susceptibility testing. Data collected included signalment, known or suspected origin of synovial contamination, synovial structures affected, antimicrobial treatment, and results of synovial fluid cytologic evaluation and bacterial culture and susceptibility testing. Horses were grouped as adults (≥ 6 months old) or foals (< 6 months old). Results of bacterial culture were positive for 34 of 70 (49%) and 18 of 61 (30%) samples from 68 adult horses and 40 foals, respectively. Gram-positive bacteria were more common in adult horses, whereas gram-negative bacteria were more common in foals. No multidrug-resistant microorganisms were identified. For adult horses, 92% (23/25) of gram-positive isolates tested with penicillin and gentamicin were susceptible to the combination. For foals, 94% (15/16) of isolates tested with penicillin, gentamicin, or both had susceptibility to 1 or both antimicrobials. Periodic review of bacterial profiles and antimicrobial susceptibility in horses with septic synovitis can help to detect early changes in bacterial pressure and antimicrobial resistance. Findings suggested that in the geographic area we serve, a combination of penicillin and gentamicin would be an effective empirical antimicrobial treatment for most horses with septic synovitis while results of bacterial culture and susceptibility are pending.