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Background: Tuber aestivum Vittad., known as black summer truffle, represents high-value food especially used as garnishment in nouvelle cuisine. The aim of this study was to investigate on the viable microbial populations associated with T. aestivum ascomata collected in different sites of Sicily and one locality of Umbria (Italy). Methods: The ripe ascomata of black summer truffles were collected from Central Italy. Cell densities of spoilage bacteria, fecal indicators, potential pathogens, yeasts, and molds were analyzed. Statistical analysis was conducted with XLSTAT software. Results: The microbiological counts of truffles ranged between 6.00 and 9.63 log Colony Forming Unit (CFU)/g for total mesophilic count and between 6.18 and 8.55 log CFU/g for total psychrotrophic count; pseudomonads were in the range 6.98-9.28 log CFU/g. Listeria spp. and coagulase-positive streptococci detected in no samples. Coagulase-negative streptococci were found in some samples with 2.11-4.76 log CFU/g levels. Yeasts and filamentous fungi were detected at consistent levels of 3.60-7.81 log CFU/g. Significant differences (p<0.01) were found between samples and also for all microbial groups. Conclusion: This study evidenced that the common brushing procedure applied for preparation of truffles is not sufficient to eliminate microbial risks for consumers. The application of an efficient decontamination treatment is strongly suggested before consumption of fresh truffles. [ABSTRACT FROM AUTHOR]