MegaSporeBiotic™ is an oral, spore-based probiotic comprised of five Bacillus spp. The study demonstrates L-alanine works well as a probiotic Bacillus adjuvant in improving intestinal health, and it also provides a solution for the practical and accurate regulation of their use as antibiotic alternatives in animal production. Improved species diversity and abundance of fecal microbiota were only observed in the group with L-alanine-treated S-2 spores ( p < 0.05). subtilis S-2 spores significantly improved the growth of the SD rats, alleviated histopathological GIT lesions, and improved the ratio of jejunal villus length to crypt depth in comparison to the B. Besides, supplement with L-alanine-treated B. subtilis S-2 spores, but not 312 spores, to decrease inflammatory factors (IL-6, IL-8, IL-1 β, TNF-α p < 0.05) and promote the expression of occludin in IPEC-J2 cells. The result indicated that L-alanine helped B. The protective effect of L-alanine-induced spore germination of the two strains was comparatively investigated both in the IPEC-J2 cell model and a Sprague–Dawley (SD) rat model challenged by enterotoxigenic Escherichia coli K99.

Consequently, after L-alanine treatment, the gray phase was largely increased under microscopy, and the expression of the germination response genes was significantly up-regulated in the B. Two strains of Bacillus subtilis, S-2 and 312, with or without a high spore germination response to L-alanine, were selected to study their morphological and genic differences induced by L-alanine through transmission electron microscopy and comparative transcriptomics analysis. The results showed the germination response was strain-specific and germinant-related, and nutrient germinant L-alanine significantly promoted the growth of strains with germination potential. In this study, based on 14 potential probiotic Bacillus strains from fermented food and feed, we detected the germination response of these Bacillus spores in relation to different germinating agents. Spores undergo strain-related germination in the gastrointestinal tract, but it is still unknown whether the probiotic function of the Bacillus depends on the germination of spores in vivo. No clear effect of high negative redox potential, which is a unique property of AlEW, on the reduction of heat resistance was recognized.Īs alternatives to antibiotics in feed, probiotic Bacillus carries multiple advantages in animal production. It was considered that the decrease in heat resistance by AlEW was resulted from the weakening of surface layer of spores by protein dissolution with alkaline substance. In case of the temperature during HPP treatment is high with lower pressure levels, bacterial spores did not reduce its heat resistance even when AlEW was combined as the pretreatment. In case of the temperature during HPP treatment was 70℃, bacterial spores did not reduce its heat resistance with lower pressuring levels.

The obtained D value of bacterial spores treated by HPP was decreased to one-eighth by the pretreatment with AlEW when compared with the control sample. An additional 2 hours duration of HPP treatment could inactivate more 2 logs of the viable bacterial spores.

However, a 3-log decrease in the viable bacterial spores was obtained by the combination of AlEW pretreatment with 1 hour of HPP treatment. No reduction of heat resistance of bacterial spores, which was defined as the spore survival by heat treatment at 80℃ for 15 min, was observed by the treatment of single HPP with 30MPa at 50℃ even for 6 hours. The effect of combined use of alkaline electrolyzed water （AlEW） on the reduction of heat resistance of bacterial spores by high hydrostatic pressure processing（ HPP） was investigated in this study.

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