In June, 1989, an outbreak of nosocomial listeriosis occurred in Costa Rica. Listeria monocytogenes was isolated from 9 ill infants 4 to 8 days old who were born after the delivery of an infant with early onset listeriosis. One nosocomial infection was fatal, 2 required mechanical ventilation and 1 resulted in hemiparesis. A higher proportion of cases than other infants born during the outbreak were delivered by cesarean section (55% vs. 24%, P = 0.04). Compared with the mothers of 36 random controls, case mothers were more often primiparous (odds ratio, 6.2, P = 0.03) or received general anesthesia before delivery (odds ratio, 4.4, P = 0.09). All infants were bathed with mineral oil from a multidose container. Culture of the oil by cold enrichment grew L. monocytogenes 4b with the same electrophoretic enzyme type as the outbreak strain. We hypothesize that aspiration of contaminated oil may have resulted in systemic listeriosis. General anesthesia may have increased the risk of aspiration. Lung tissue from the infant who died showed lipid-laden macrophages consistent with oil aspiration and had evidence of L. monocytogenes DNA detected by polymerase chain reaction. This is the first nosocomial outbreak of listeriosis in which a common source suggested epidemiologically was microbiologically confirmed. The high attack rate (greater than 200 times the United States rate of perinatal listeriosis) emphasizes the susceptibility of healthy neonates to L. monocytogenes. The results of our study led to the discontinuation of the use of mineral oil for bathing neonates in Costa Rica.