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Blood cultures (BCs) are essential microbiologic tests, but blood culturing diagnostic stewardship is frequently poor. We aimed to study the process-related failures and to evaluate the effect of an emergency department (ED) intervention on BCs collection practices and yield. We implemented an ED-quality improvement intervention including educational sessions, phlebotomists addition, promoting single-site strategy for BC-collection and preanalytical data feedback. BC-bottles collected, positive BCs, blood volumes and documentation of collection times were measured, before (December 2021-August 2022) and after (September 2022-July 2023) intervention. Results were corrected to hospitalizations admissions or days. We used interrupted-time series analyses for comparisons. A total of 64,295 BC bottles were evaluated, 26,261 before and 38,034 postintervention. The median ED-BCs collected per week increased from 88 to 105 BCs (P <.0001), resulting from increased early sampling (P =.0001). Solitary BCs decreased (95%-28%), documented times increased (2.8%-25%), and average blood volume increased (3 mL to 4.5 mL) postintervention. Community-onset Bloodstream infections (BSIs) increased (39.6-52 bottles/1,000 admissions, P =.0001), while Health care–associated BSIs decreased (39-27 bottles/10,000 days, P =.0042). Contamination rates did not change. An ED-focused intervention based on the education sessions and single-site strategy improved culturing stewardship and facilitated the early identification of BSI without an increase in contamination. • Diagnostic stewardship of blood cultures is frequently poor. • ED intervention was based on switching to single-site strategy for blood culturing. • Admission blood cultures number increased, and solitary blood cultures reduced. • Community-onset bloodstream infections detection increased after the intervention. • There was no increase in blood-culture contamination rates. [ABSTRACT FROM AUTHOR]