In current clinical practice, in case of symptoms-based suspicion of diabetic foot infection, immediate empiric antibiotic therapy is recommended. Prevailing guidelines do not provide region-specific therapy schemes. To validate existing recommendations, there is an urgent need for a report on diabetic foot infection microbiota patterns in Central Europe. This study aimed to describe diabetic foot infections microbiota and its antibiotic susceptibility in Poland. We conducted a single-center descriptive study at the General Surgery Department, Siedlce Hospital, Poland. Data for all patients diagnosed with diabetic foot infection between January 1, 2015, and December 31, 2016, and corresponding antimicrobial susceptibility tests were extracted. A total of 54 patients were included in the study, with a total of 102 microbiological samples. Among 81 positive samples, 77.1% of the isolated bacteria were Gram-negative. A total of 93.4% of the isolates were facultatively anaerobic bacteria. No obligatory anaerobic bacteria and no yeasts were isolated. Facultatively anaerobic, Gram-negative bacteria, mainly Proteus spp and Escherichia coli, were the most common organisms cultured in diabetic foot infections. This study suggests that the currently implemented treatment recommendations might not be adequate in Poland.