Introduction Chronic osteomyelitis (COM) is a devastating infection requiring a multidisciplinary approach, including radiology, microbiology, pathology, and orthopaedic surgery to treat. The present study analysed the bacterial profile causing chronic osteomyelitis and their antibiogram in our region. Patients and methods This prospective study was done on a consecutive group of patients who underwent surgical debridement for long bone COM. Three to six deep tissue samples were collected during the index debridement for microbiology and one sample for histopathology. Antimicrobial sensitivity testing used an automated bacterial identification system. Gram stain was used to identify the bacteria type from its size, shape, and arrangement of bacterial growth. Results Intra-operative deep tissue and bone specimens accurately identified causative bacteria in 84.8% of patients. Gram-ve bacilli (GNB) were the most common causative organisms in 51.6% of all growing samples (36.4% isolated G-ve and 15.2% mixed with G + ve). Thirty-three patients (30 males/three females) were included; the mean age at index debridement surgery was 37.1 years. Half of the cohort had no metalwork. The aetiology of COM was post-operative infection in half of the patients. Conclusion There may be concerning features in our patients’ aetiologies and causative organisms; closed fractures turn into COM postoperatively, several unsuccessful attempts, delayed index debridement, and more GNB. Plans need to be applied to break the cycle and improve outcomes.