If a bone or joint infection is suspected, perioperative antibiotic prophylaxis is frequently withheld until the intraoperative microbiological sampling has been performed. This practice builds upon the hypothesis that perioperative antibiotics could render culture results negative and thus impede tailored antibiotic treatment of infections. We aimed to assess the influence of antibiotic prophylaxis within 30 to 60 minutes before surgery on time to positivity of microbiological samples and proportion of positive samples in Cutibacterium acnes bone and joint infections. Patients with at least one positive C. acnes sample between January 2005 and December 2015 were included and classified as 'infection' if at least 2 samples were positive, otherwise they were considered a 'contamination'. Kaplan-Meier curves were used to illustrate time to culture positivity. We found 64 cases with a C. acnes infection and 46 classified as a C. acnes contamination.Application of perioperative prophylaxis significantly differed between the 'infection' and 'contamination' group (72.8% versus 55.8%, p<0.001). Within the 'infection' group, we found no difference in time to positivity between those who had or had not received a perioperative prophylaxis (7.07 days (95% CI 6.4-7.7) vs. 7.11 days (95% CI 6.8-7.5), p=0.3). Also, there was no association between the proportion of sample positivity and the application of perioperative prophylaxis (71.6% versus 65.9%, p=0.39). Since perioperative prophylaxis did not negatively influence the microbiological yield in C. acnes infections, routine antibiotic prophylaxis can be routinely given to avoid surgical site infections.