Background: Current evidence suggests that more than half of all antimicrobials are used in the sector of food-producing animals, thus constituting a major risk factor for development and spread of antimicrobial resistance (AMR). Methods: This cross-sectional study assessed the knowledge, attitudes, and practices regarding antimicrobial resistance (AMR) and antimicrobial use (AMU) among veterinarians (n = 26) working with food-producing animals and operators (n = 165) of establishments that keep food-producing animals, across all districts of Cyprus between October and December 2024. Results: Most veterinarians demonstrated sufficient knowledge regarding AMR and AMU; however, certain knowledge gaps were identified. There was a general trend toward desired responses aimed at reducing AMU. Despite this, the level of responses advocating for restrictions on the use of specific priority antimicrobials and broad-spectrum antibacterials was unsatisfactory. Over half of veterinarians prescribed Category B substances. Furthermore, there was no significant association between the use of “restricted” antibiotics and veterinarians’ level of knowledge nor between antibiotic use and the practice of microbiological culture and susceptibility testing. Among operators, positive attitudes were significant predictors of implementing of good practices. Additionally, operators who had contracts with a veterinarians were more likely to follow good practices. Conclusions: There is a need for further education on AMR in the veterinary sector in the Republic of Cyprus. Raising awareness among animal producers, is also crucial, along with enforcing a policy on antimicrobial use. Comprehensive governance involving all stakeholders must be implemented to address AMR more effectively.