The authors characterized the genera of mycelial fungi found in samples of expressed human milk received through home collection by the Human Milk Bank of the Instituto Fernandes Figueira in Rio de Janeiro. A total of 821 samples of expressed human milk were taken randomly from bottles collected at home by the milk donors themselves and were investigated for molds, yeasts, and mesophilic microorganisms. The analyses showed the occurrence of molds and yeasts in 43 (5.2%) of the samples, with counts reaching 103CFU/ml. Some 48 strains of mycelial fungi were identified by standard laboratory techniques, including: Aspergillus Niger group (6.3%), Aspergillus sp. (4.2%), Paecilomyces sp. (12.6%), Penicillium sp. (60.4%), Rhizopus sp. (2.0%), and Syncephalastrum sp. (14.5%). The authors discuss the importance of donor hands' asepsis prior to collecting human milk.