To assess heat tolerance of microencapsulated L. reuteri DSM 17938 and L. plantarum 48M cells, bacteria survival was tested against four time-temperature combinations: 60 °C for 3 min, 70 °C for 2 min, 80 °C for 2 min, 80 °C for 5 min. Samples were prepared as follows: free or microencapsulated cells were 10-fold diluted in MRS Broth and sealed in glass capillary tubes (Ø 2 mm). The tubes were then placed in a thermostatic water bath and treated at the conditions cited above. At the end of the period, glass tubes were removed and rapidly cooled under running tap water.

Aliquots were sampled before and after thermal treatments and counted by serially diluting 1 mL aliquots of the test material, pour plating on MRS Agar and incubating at optimal growth temperature for the tested strain for 48 h. Results were expressed in terms of surviving percentage as Nt/N0 × 100, where N0 and Nt are cell loads expressed in CFU/mL before and after heat exposure, respectively. Three independent experiments were carried out.

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