Reuterin production from L. reuteri DSM 17938 cells was tested by an antagonistic deferred agar spot test [26]. Ten microlitres of sample (free or microencapsulated cells) were spotted on plates of MRS Agar supplemented with 250 mM glycerol and incubated in anaerobic conditions at 37 °C for 24 h. Then, plates were covered with 10 mL of TSB supplemented with 0.75% agar and 0.5% yeast extract and seeded with 2% of an overnight culture of the indicator strain (Escherichia coli 32, Hafnia alvei 53M or Pseudomonas fragi 25P). After the incubation at optimal growth temperature for 24 h, the antimicrobial activity was observed as an inhibition zone of the indicator organism around the sample.

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