Background and objectives Intra-abdominal infections (IAIs) are associated with significant morbidity and mortality. Pathogenic isolates and emerging resistance to commonly used antimicrobials have been a matter of concern in IAIs. In the present study, bacteriological profile and antimicrobial susceptibility of isolates from IAIs were investigated. Materials and methods A total of 145 samples (ascitic fluids, n = 56; bile, n = 20; and pus, n = 36) were collected from suspected IAI of patients reporting to the hospital and cultured. Identification of the isolates was done using standard identification protocol. Antimicrobial susceptibility was performed by Kirby-Bauer disk diffusion method and interpretation was done according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. Results Of 145 samples, 112 were culture positive and 33 were sterile. Gram-negative organisms (n = 85) outnumbered the Gram-positive organisms (n = 27). Among the Gram-negative organisms, Escherichia coli (n = 31) was the most commonly isolated organism followed by Klebsiella sp. (n = 19), Acinetobacter sp. (n = 14), Pseudomonas sp. (n = 10), Proteus sp. (n = 5), Citrobacter sp. (n = 3), and Enterobacter sp. (n = 3). Among the Gram-positive bacteria, the most common organism was Staphylococcus aureus (n = 19) followed by Enterococcus faecalis (n = 8). Gram-negative bacilli showed significant resistance to almost all of the commonly used antibiotics. The rate of methicillin-resistant S. aureus (MRSA) was 36.84%. Conclusion Prompt starting of empirical antimicrobials based on the local susceptibility pattern, followed by modification of treatment in accordance with the antimicrobial susceptibility report can significantly reduce the morbidity and the mortality associated with IAIs. How to cite this article Garg R, Singh VA. Bacteriological Profile and Antimicrobial Susceptibility Pattern of Intra-abdominal Infections: A Study from a Tertiary Care Hospital of North India. J Gastro Infect 2017;7(1):21-25.