Background: Antibiotic resistance is a major global threat to mankind. Inappropriate antibiotic use has been identified as one of the greatest contributors to antibiotic resistance. There is currently a paucity of data on antibiotic use in aged care facilities, globally and in the Australian setting. Aim: To investigate the current antibiotic prescribing practices and use in aged care facilities in the Riverina, New South Wales (NSW), and metropolitan Australian Capital Territory (ACT). Method: Care managers of aged care facilities in the ACT and Riverina, as well as pharmacists who service aged care facilities in regional NSW and ACT, were asked to complete a questionnaire relating to antibiotic use in aged care facilities. Data obtained were subjected to statistical analysis. Results: Most facilities in the ACT (85%) and NSW (90%) had some form of protocol in place for the prescribing and obtaining of antibiotics for residents. The most commonly prescribed antibiotic across participating facilities was cephalexin and the most common indication for antibiotic use was urinary tract infection. No statistically significant differences in antibiotic use and antibiotic prescribing practices were observed between metropolitan ACT and regional NSW aged care facilities. There was a statistically significant association between facilities with residents taking antibiotics for ongoing conditions and those that have experienced situations in the past 12 months in which the initially prescribed antibiotic did not work and another had to be used (p = 0.004944). Conclusion: The data from this study can inform further studies that may be used to inform the development of antimicrobial stewardship initiatives in aged care facilities.