Disseminated candidiasis is the most common nosocomial fungal infection, and Candida albicans has been reported to account for 50% to more than 70% of cases of invasive candidiasis. However, recent reports have also suggested the emergence of infections caused by non-albicans species. In addition, less-common pathogenic yeasts (Malassezia, Trichosporon, Rhodotorula, Debaryomyces and Pichia) have recently been reported, with increased frequency, as causes of nosocomial infections with high mortality. This article describes two cases of fungemia caused by Pichia anomala in newborns that occurred in an intensive care unit (ICU), in November 2004 at the Instituto da Criança (Pediatric Institute) of the Hospital das Clínicas of the School of Medicine, São Paulo University, Brazil. The principal factors related to virulence (proteinase and phospholipase) and the susceptibility of the isolated strains to antifungal agents were also evaluated, and the biotype of each strain was determined through the use of an epidemiological marker (killer biotype).