Inappropriate use of antibiotics in humans and animals contributes to decreased antimicrobial susceptibility in bacteria of medical importance. Resistant bacteria being transferred from animals to humans are causing public health concern. In-person interviews were conducted with 20 dairy farmers in rural counties of South Carolina to determine farmers' knowledge and attitudes about prudent antibiotic use among livestock. Four focus groups (n = 22) were also conducted to ascertain farmers' specific information needs about proper antibiotic use. Survey results showed that participants (100%) typically determined a need for antibiotic treatment using symptom assessment and reported following some form of operating procedures regarding administration of antibiotics. Few farmers (32%) had actual written antibiotic protocols. Preferred information sources about antibiotics were veterinarians (100%) and other dairy farmers (50%). Most farmers (86%) were not concerned that overuse of antibiotics in animals could result in antibiotic resistance among farm workers. Qualitative analysis of focus groups revealed significant barriers to following proper antibiotic procedures including limited finances and lack of time. The need for bilingual educational resources for Hispanic/Latino dairy workers was expressed. Desired formats for educational materials were posters, flowcharts, videos, and seminars. Education of South Carolina dairy farmers by veterinarians and public health professionals on the appropriate use of antibiotics in dairy cattle is needed to ensure antibiotic effectiveness in both animals and humans.