Abstract Background Although antimicrobial stewardship (AS) personnel are faced with occupational stressors that may contribute to burnout, little is known about the degree to which they experience this work-related syndrome. Our objective was to characterize the prevalence of, and identify factors associated with, burnout among AS personnel practicing in United States (US) hospitals. Methods We conducted a cross-sectional survey in October-December 2021. AS personnel (physicians and pharmacists) were approached via email through four US-based professional organizations. Respondents self-administered a questionnaire measuring demographics, AS program structure, resources, and organizational climate. Burnout was measured using the Maslach Burnout Inventory, a 22-item validated instrument with three subscales. Burnout was defined as a dichotomous outcome based on the presence of high scores on the emotional exhaustion subscale along with either a high depersonalization score or low professional accomplishment score. Descriptive statistics and logistic regression analyses were performed. Results A total of 259 AS personnel completed the questionnaire. The majority of respondents were pharmacists (n=201; 78.2%), female (n=166; 64.3%) and worked in a teaching hospital (n=212, 84.1%). The median age was 36 (IQR, 32-41). Thirty-six percent (n=94 of 259) of respondents met the definition for burnout. There were no significant differences in burnout by respondent demographics or professional role. Significant drivers of burnout (P< 0.05) identified in univariable regression modeling included feeling as if AS is not an institutional priority, that hospital leadership is unsupportive of AS, prescriber resistance to AS recommendations is common, having unsupportive infectious diseases or pharmacy colleagues, the AS team does not work well together, and AS work is not intellectually stimulating. Conclusion Over one third of AS personnel in our sample met predefined standardized criteria for burnout. Contextual aspects of the work environment, including leadership, unsupportive peer climate and conflict were associated with burnout. There is a need to address factors that contribute to burnout in AS personnel to ensure an engaged workforce and to minimize turnover. Disclosures Jason Newland, MD, AHRQ: Grant/Research Support|Merck: Grant/Research Support|NIH: Grant/Research Support|PEW Charitable Trust: Grant/Research Support|Pfizer: Grant/Research Support.