The North American (NA) porcupine (Erethizon dorsatum) is a rodent species with specialized hair structures called quills designed to detach and penetrate into tissues of any human or animal coming into contact with them. The objective of this study was to characterize the fungal flora of the quills in the region of the rosette in wild NA porcupines to further define health risks to NA porcupines and any animal coming into contact with the quills. A total of 17 adult NA porcupines were sampled, and fungal culture was performed. Fungal organisms were cultured from 15 (88.2%) of 17 samples. Thirty-three isolates of 10 different fungal genera were cultured. The most frequently isolated fungi were Lodderomyces elongisporus (n = 7, 41.2%), Candida spp. (n = 3, 17.6%), and Penicillium spp. (n = 2, 11.8%). Eleven (64.7%) individuals grew multiple fungal organisms. In humans and animals quilled by porcupines, fungal culture should be considered in cases of infection, and if isolates resembling Candida spp. are isolated, matrix-assisted laser desorption–ionization time of flight or molecular methods are necessary to rule out L. elongisporus.