Background: Employing standard precautions for infection prevention is increasingly concerned in operating theater. The KAP of Anesthetists plays a vital role in minimizing the risk of infection in surgical patients. Objective: To assess knowledge, attitude and practices of Anesthetists' on standard precaution of infection prevention. Methodology: Facility based cross sectional study was conducted in JUTH from MAY 08 -22/2015. Data was collected from all Anesthetists using pretested self-administered questionnaire. The knowledge level, attitude and practice of respondents were analyzed as predefined scale levels using scientific calculator and computer. Finally the result was presented as total percentage using tables and descriptive statements. Result: Eighteen questionnaires were distributed and 17 (94.44%) were found to be valid and analyzed. Based on grading of knowledge level 14 (82.35%) of them has good knowledge while the rest of 3 (17.64%) have poor knowledge. Among 17, 14 (82.35%) attitude score was > 15 and 3 (17.64%) score was below 15. Therefore 14 (82.35%) Anesthetists have favorable attitude and 3 (17.64%) have unfavorable attitude on standard precaution. Out of 17, 4 (23.52%) was exposed and 3 (75%) of them wash with soap and water and use hand antiseptics as immediate measure after exposure. Conclusion and Recommendation: The majority of Anesthetists had poor practice of hand washing before and between every patient they intended to care. So a continual training program should be arranged for Anesthetists to follow the trend of hand washing with standard protocols for hand hygiene in the health care environment.