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Antimicrobial use contributes to the development of emergence and dissemination of antimicrobial-resistant bacteria among intensive care unit (ICU) patients. There are few published data on antimicrobial use in neonatal (NICU) and pediatric ICU (PICU) patients.Personnel at 31 Pediatric Prevention Network hospitals participated in point prevalence surveys on August 4, 1999 (summer) and February 8, 2000 (winter). Data collected for all NICU and PICU inpatients included demographics, antimicrobials and indications for use and therapeutic interventions.Data were reported for 2647 patients in 29 NICUs (827 patients in summer; 753 in winter) and 35 PICUs (512 patients in summer; 555 in winter). PICU patients were more likely than NICU patients to be receiving antimicrobials on the survey date [758 of 1070 (70.8%) versus 684 of 1582 (43.2%), P < 0.0001]. NICU patients were receiving a higher median number of antimicrobials (2versus 1, P < 0.0001). The most common agents among NICU patients were gentamicin, ampicillin and vancomycin; the most common agents among PICU patients were cefazolin, vancomycin and cefotaxime. Use of aminoglycosides, aminopenicillins and topical antibacterials was significantly more common in NICU patients; first, second and third generation cephalosporins, extended spectrum penicillins, sulfonamides, fluoroquinolones, antianaerobic agents, systemic antifungals and systemic antivirals were more common in PICU patients.This is the first U.S. national multicenter description of antimicrobial use in NICUs and PICUs and demonstrates the high prevalence of antimicrobial use among these patients. Assessment strategies targeting antimicrobial use in pediatrics are needed.