AimsBacillus probiotics recently gained attention due to the production of resistant cells. The invitro probiotic potentials and safety assessment were evaluated for three Bacillus strains obtained from traditional pickle. Methods and ResultsThree bacterial strains designated as 437F, 1630F and 1020G were isolated from a traditional pickle and identified as members of the genus Bacillus. The novel strains showed high acid and bile tolerance. They exhibited antagonistic activity against various pathogens. Antioxidant activity, auto- and co-aggregation ability as well as their surface hydrophobicity and attachment capacity to the Caco-2 cells were in the range of other well-known probiotic strains. They were susceptible to various antibiotics. The enterotoxin (HBl and NHe), cytotoxin (Cytk1) and emetic (Ces) genes were not detected based on PCR assay. They were not toxic against HT-29 cells. ConclusionWith respect to their characteristics and safety aspect, these Bacillus species may have potential to consider as probiotics for animal and/or human applications. Significance and Impact of the StudyNondairy-fermented foods are interesting sources for isolation of novel probiotics. Identification of novel Bacillus strains with remarkable probiotic potentials would increase their contribution in food/feed and pharmaceutical industries.