Introduction: Polymicrobial biofilm in chronic diabetic foot wounds is a major challenge for the treatment and lead to several complications, including increased morbidity, mortality, and limb loss.   Methodology: In this study, a spectrum of bacterial and fungal isolates was carried out in 66-foot ulcer samples from diabetic patients. All bacterial isolates were identified by conventional culture methods and biochemical identification tests. Filamentous fungi and yeasts were identified morphologically and by staining methods.   Results: Among 66 samples, 62 (94%) samples were culture-positive, and four samples (6%) were culture-negative by the conventional culture method. Polymicrobial growth was found in 11 samples (17%) and monomicrobial in 51 wound samples (77%). Both Gram-positive and Gram-negative bacterial species were isolated. The number of Staphylococcus aureus isolates was more (61%).  Fungi were isolated in 4 patients, and all the isolates were identified as Candida tropicalis. Several clinical complications were found including hypertension (47%), osteoarthritis (6%), rheumatoid arthritis (8%), cardiovascular disease (19%), peripheral vascular disease (20%), amputation (35%), foot cramps (71%) and edema (8%).   Conclusion: Monomicrobial growth was predominant than polymicrobial growth, and S. aureus was the predominant pathogen isolated. Adequate treatment management of diabetic foot is needed to reduce the risk of amputation.