*2.10. Microbiological Analysis*

Samples of a specific weight were pacified to decimal serial dilutions in Ringer's solution (Sigma-Aldrich, Milan, Italy), and 25 g of the fruit samples were homogenized in a flask containing 225 mL of Ringer's solution (Sigma-Aldrich, Milan, Italy) using the Bag-Mixer 400 stomacher (Interscience, Saint Nom, France) for 2 min at the highest speed (blending power 4). All test tubes of serial dilutions used contained Ringer's solution (9 mL). Peptone Dextrose agar (PDA) was used for all plate media. A total of 1 mL of the bacterial suspension was pipetted into a dilution tube containing 9 mL of Ringer's solution. This tube was vortexed, and 1 mL of this volume was removed and placed into a second dilution tube containing 9 mL of Ringer's solution. This process was repeated until the sample was sufficiently diluted. These PDA plates are prepared in three replicates, and 100 mL of this suspension was added to the plates and repeated for every tube in the dilution series. The different microbial groups were investigated as follows: total mesophilic microorganisms (TMM) on plate count agar (PCA) incubated at 37 ◦C for 48 h and total yeasts and molds (TYM) on PDA supplemented with chloramphenicol (0.1 g/L) to avoid the growth of bacteria then incubated for 48 h at 25 ◦C. Plate counts were achieved by the spread plate method [56] by inoculating 100 μL from each sample's suspension of appropriate dilution. All media were supplied from Oxoid (Milan, Italy). At each collection time, the microbiological counts were performed in triplicate.

### *2.11. Statistical Analysis*

Data were subjected to statistical analysis for calculation of means, variance and standard error using CoStat Software Program Version 6.303 (CoHort Software, Monterey, CA, USA) using one-factor analysis of variance (ANOVA, general linear model), followed by Duncan multiple range test for *p* < 0.05 [57] was used to test the differences among treatments.
