The rise of multidrug-resistant bacterial species in hospitals becomes a global challenge for surgeons who treat healthcare-associated infections. This study aimed to identify the pathogens involved in surgical site infections (SSI) as well as the prevalence of antibiotic resistant bacteria in the Nilgiris region. A hospital-based retrospective study was conducted for three years, at Microbiology Laboratory, the Govt. Medical College Hospital, where the clinical samples were collected, cultured, and identified. Antibiotic susceptibility was assessed using Kirby Bauer’s disc diffusion method. Out of 513 pus samples (from SSI), 242 (47%) have shown positive microbial growth. These isolates were evaluated for antimicrobial resistance using 20 antibiotics belonging to different groups. Staphylococcus aureus was found to be more prominent (69%), followed by Enterococcus species (14.5%) and Streptococcus species (10.3%). Other species like Proteus species, Klebsiella species, Escherichia coli, and Pseudomonas aeruginosa account for less than 2%. These results clearly indicate that Staphylococcus aureus was the leading cause of surgical site infections. Among the antibiotics studied, Staphylococcus aureus was found to be more resistant to Penicillin G (84%) followed by Ampicillin (23%). The high rate of antibiotic resistance highlighted the need for an antibiotic policy that encourages more rational use of antibiotics.