Purpose To identify temporal changes in antimicrobial resistance of ocular surface bacteria isolated from clinically symptomatic equine eyes in the South West of the UK. Study design Retrospective. Methods Clinical and laboratory records of horses treated for suspected bacterial ocular surface disease (ulcerative and non‐ulcerative) at a single facility between January 2011 and December 2019 were reviewed. Cases were included if they underwent ocular surface sampling, aerobic bacterial culture, and antimicrobial susceptibility testing. Cases were split into two time periods based on when sampling occurred: “early” (2012‐2015) and “late” (2016‐2019) to enable identification of temporal trends in resistance to chloramphenicol, gentamicin, fusidic acid, neomycin, cloxacillin, ofloxacin, and polymyxin B. Results A total of 125 samples from 110 horses were included in analyses. Culture‐positive isolates were identified in 76/110 (60.8%) samples. Principal isolates included Staphylococci spp. (n = 45; 64.3%), Streptococci spp. (n = 14; 20%), and Enterobacter spp. (n = 11; 15.7%). There was a significant increase in resistance to chloramphenicol over time (P = .007) and a decrease in resistance to ofloxacin that approached significance (P = .059). Chloramphenicol (100%) and gentamicin (85.7%) had the highest overall in‐vitro efficacy during the early and late periods, respectively. There was no significant difference in the type of bacteria isolated across the two time periods. Conclusions These results suggest a potential increase in resistance to chloramphenicol among bacteria isolated from the ocular surface of horses in the South West UK, reinforcing the value of surveillance to guide the empirical use of antimicrobials.