The membrane lipid ergosterol is found almost exclusively in fungi, and is frequently used by environmental microbiologists as an indicator of living fungal biomass, based on the assumption that ergosterol is labile, and therefore rapidly degraded after the death of fungal hyphae. We studied the degradation of ergosterol in environmental samples without living fungi. Under the conditions used in this study, ergosterol was very stable both when added as a pure compound and when associated with dead fungi. The decrease of ergosterol was at most 34% during 2 months when protected from sunlight. Presence of a natural bacterial assemblage did not enhance degradation over this time period, as compared to sterile controls. However, photochemical degradation was significant, and led to a 43% decrease of in ergosterol content during 24 h. These results suggest that ergosterol should be used cautiously as a biomarker for living fungi.