Microbiological surveillance in the operating theatres is one of the most important strategies for determining potential sources of contamination and putting infection control strategies into effect. Regular monitoring establishes the environment conducive to surgical site infections and helps in formulating necessary interventions. Prior studies have shown that the increased levels of microbial contamination in the surgical environment necessitate strict microbiological monitoring. The present study was conducted to identify bacterial colonization of surfaces and equipment in the OTs and to determine the microbial contamination of air in the OTs of a tertiary care hospital.