Ambient particles comprise approximately 25% of fungal spores, which cause adverse health outcomes such as respiratory diseases, allergy, and infection. In this study, we investigated temporal variations and distributions of ambient fungal spores in an urban area of the Taipei metropolis for over 1 year. A Burkard 7-day volumetric spore trap was used to collect air samples. Samples collected daily were stained, counted, and identified on the basis of morphological characteristics. The associations between fungal spores and environmental parameters were then evaluated through multiple regression analysis. Daily monitoring data revealed a large variation in fungal spore concentrations. Specifically, fungal spores peaked during summer months (June–August) and declined during winter months (December–early March); moreover, the average concentration of total fungal spores was 3,607.97 ± 3,181.81 spores m–3. Ascospores were the most prevalent taxon that was recovered from the samples, followed by basidiospores, Aspergillus/Penicillium, and Cladosporium. Multiple regression analysis revealed that meteorological parameters were the main predictors of fungal concentrations. Temperature, wind speed, and humidity were consistently correlated with total fungi and major fungal taxa, and sunlight had a negative association with ascospores. Among the atmospheric pollutants, particulate matter with an aerodynamic diameter ≤ 10 µm (PM10) and ozone were positively associated with fungal spores. Carbon monoxide (CO) at lag day 1 had a negative association with basidiospores. This is the first study to characterize daily concentrations and determinants of ambient fungal spores in an urban area of Taipei metropolis. The obtained data can be used to evaluate the health impact of fungal spore exposure on the residents of the Taipei metropolitan area.