This study characterizes the oral fungal microbiome (mycobiome) in 20 healthy individuals using a novel multitag pyrosequencing approach. The researchers identified 74 culturable and 11 non-culturable fungal genera in the oral cavity, with 101 species in total. Candida species were the most frequently isolated (75% of participants), followed by Cladosporium (65%), Aureobasidium, and Saccharomycetales (50% each), Aspergillus (35%), Fusarium (30%), and Cryptococcus (20%). Four of these genera are known to be pathogenic in humans. The study also found that 15 genera were present in at least 20% of the samples. The oral mycobiome showed differences in distribution based on gender and ethnicity, with White males and Asian males clustering separately, while Asian and White females clustered together. This is the first study to identify the "basal mycobiome" of healthy individuals, providing a foundation for further research on the oral mycobiome in health and disease. The findings highlight the diversity of the oral fungal microbiome and its potential role in health and disease. The study also emphasizes the importance of non-culturable fungi in the oral cavity, which may play a significant role in the oral environment. The results suggest that the oral mycobiome is complex and varies among individuals, with implications for understanding the role of fungi in oral health and disease.This study characterizes the oral fungal microbiome (mycobiome) in 20 healthy individuals using a novel multitag pyrosequencing approach. The researchers identified 74 culturable and 11 non-culturable fungal genera in the oral cavity, with 101 species in total. Candida species were the most frequently isolated (75% of participants), followed by Cladosporium (65%), Aureobasidium, and Saccharomycetales (50% each), Aspergillus (35%), Fusarium (30%), and Cryptococcus (20%). Four of these genera are known to be pathogenic in humans. The study also found that 15 genera were present in at least 20% of the samples. The oral mycobiome showed differences in distribution based on gender and ethnicity, with White males and Asian males clustering separately, while Asian and White females clustered together. This is the first study to identify the "basal mycobiome" of healthy individuals, providing a foundation for further research on the oral mycobiome in health and disease. The findings highlight the diversity of the oral fungal microbiome and its potential role in health and disease. The study also emphasizes the importance of non-culturable fungi in the oral cavity, which may play a significant role in the oral environment. The results suggest that the oral mycobiome is complex and varies among individuals, with implications for understanding the role of fungi in oral health and disease.