Background: Post-operative sepsis remains a significant cause of morbidity and mortality in surgical patients. While the importance of timely antibiotic administration in sepsis is established, the specific impact of beta-lactam timing on outcomes in post-operative sepsis has not been well characterized across different surgical populations. Methods: We conducted a multi-center retrospective cohort study of 150 patients (75 orthopedic and 75 general surgery) who developed sepsis within 30 days following surgery at five tertiary care hospitals between 2020-2023. The primary exposure was time from sepsis recognition to beta-lactam administration, categorized as ≤1 hour, 1-3 hours, 3-6 hours, and >6 hours. The primary outcome was 30-day mortality. Secondary outcomes included ICU and hospital length of stay, time to resolution of organ dysfunction, and functional status. Results: Median time to beta-lactam administration was 2.4 hours (IQR 1.3-4.7), with only 18.7% of patients receiving antibiotics within 1 hour. Thirty-day mortality was 19.3% overall, with a clear dose-response relationship across timing categories: 7.1% (≤1 hour), 15.3% (1-3 hours), 23.8% (3-6 hours), and 38.1% (>6 hours). In multivariable analysis, each hour delay in beta-lactam administration was associated with a 20% increase in the odds of 30-day mortality (adjusted OR 1.20, 95% CI 1.09-1.33, p<0.001). This association was stronger in general surgery patients (adjusted OR 1.28, 95% CI 1.13-1.46) compared to orthopedic surgery patients (adjusted OR 1.12, 95% CI 1.01-1.24). Delayed administration was also associated with prolonged ICU stay, hospital length of stay, and slower resolution of organ dysfunction. In the subset of patients with plasma drug measurements, achieving therapeutic concentrations within 2 hours was associated with improved survival (88.2% vs. 62.5%, p=0.027). Conclusions: In post-operative sepsis, each hour delay in beta-lactam administration is associated with a 20% increase in 30-day mortality, with a more pronounced effect in general surgery patients. These findings highlight the critical importance of early recognition and prompt antibiotic therapy for post-operative sepsis and suggest potential benefits from specialized sepsis protocols for surgical patients
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