Background: Routine anaerobic blood culture collection in febrile children is controversial, as clinicians try to account for the severe but relative infrequency of anaerobic bacteremia. Furthermore, clinical and laboratory practice variation among institutions may lead to potentially inaccurate epidemiological data. Our goal was to assess blood culture practices in pediatric patients throughout an international network of hospitals in industrialized countries. Methods: We conducted a survey of current clinical and laboratory practice patterns in a convenience sample of international institutions participating in 6 pediatric emergency research networks in the US, Canada, Europe, Australia, and New Zealand. A lead clinician at each institution queried institutional practices from the emergency department, pediatric intensive care unit, and oncology medical directors. The microbiology director at each institution completed the laboratory survey. Results: Sixty-five of 160 (41%) invited institutions participated in the survey. Routine anaerobic blood cultures are collected in 30% of emergency departments, 30% of intensive care units, and 48% of oncology wards. Reasons for restricting anaerobic culture collection included concerns regarding blood volume (51%), low pretest probability (22%), and cost-effectiveness (16%). The most common reasons institutions allow for selectively obtaining anaerobic cultures are clinical suspicion (64%) and patients who are immunosuppressed (50%). The microbiology survey showed variation in systems, although most use the BACTEC™ culture system and MALDI-TOF for organism identification. Conclusions: There is broad variation in anaerobic blood culture practices among a network of pediatric hospitals in industrialized countries.