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Background. Nosocomial infections result in considerable morbidity and mortality among neonates in high-risk nurseries (HRNs). Purpose. To examine the epidemiology of nosocomial infections among neonates in level III HRNs. Methods. Data were collected from 99 hospitals with HRNs participating in the National Nosocomial Infections Surveillance system, which uses standard surveillance protocols and nosocomial infection site definitions. The data included information on maternal acquisition of and risk factors for infection, such as device exposure, birth weight category (≤1000, 1001 through 1500, 1501 through 2500, and >2500 g), mortality, and the relationship of the nosocomial infection to death. Results. From October 1986 through September 1994, these hospitals submitted data on 13 179 nosocomial infections. The bloodstream was the most frequent site of nosocomial infection in all birth weight groups. Nosocomial pneumonia was the second most common infection site, followed by the gastrointestinal and eye, ear, nose, and throat sites. The most common nosocomial pathogens among all neonates were coagulase-negative staphylococci, Staphylococcus aureus, enterococci, Enterobacter sp, and Escherichia coli. Group B streptococci were associated with 46% of bloodstream infections that were maternally acquired; coagulase-negative staphylococci were associated with 58% of bloodstream infections that were not maternally acquired, most of which (88%) were associated with umbilical or central intravenous catheters. Conclusions. Bloodstream infections, the most frequent nosocomial infections in all birth weight groups, should be a major focus of surveillance and prevention efforts in HRNs. For bloodstream infections, stratification of surveillance data by maternal acquisition will help focus prevention efforts for group B streptococci outside the HRN. Within the nursery, bloodstream infection surveillance should focus on umbilical or central intravenous catheter use, a major risk factor for infection.