Fifty species of guanophilic (bat guano-loving) fungi were isolated from field-collected samples within three caves in southwestern Puerto Rico; most were mitosporic fungi (23 species). The caves studied were Cueva La Tuna (Cabo Rojo), Cueva de Malano (Sistema de Los Chorros, San Germán), and Cueva Viento (El Convento Cave-Spring System, Guayanilla-Peñuelas). The most conspicuous fungus by far was the zygomycete Circinella umbellata (Mucorales). Circinella umbellata dominated the bat guano incubation chambers (Petri dishes lined with sterile filter paper moistened with sterile water) at ambient laboratory conditions. Nineteen species of basidiomycetes (e.g., Ganoderma cf. resinaceum, Geastrum cf. minimum, Lepiota sp., Polyporus sp., Ramaria sp.) and three species of ascomycetes (Hypoxylon sp., Xylaria anisopleura, and X. kegeliana) were also recorded. They were found on soil, rotting leaves, bark and rotting wood, buried in bat guano located below natural skylights or sinkholes.