Bacillus subtilis spores are being used for oral bacteriotherapy and bacterioprophylaxis of gastrointestinal disorders in both humans and animals. Since B. subtilis is an aerobic saprophyte, how spores may benefit the gut microbiota is an intriguing question, since other probiotics such as Lactobacillus spp. which colonize the gut are anerobes. As a first step in understanding the potential effects of ingesting spores, we have characterized five commercial products. An extensive biochemical, physiological, and phylogenetic analysis has revealed that four of these products are mislabeled. Moreover, four of these products showed high levels of antibiotic resistance.Probiotics, or "friendly bacteria," are becoming increasingly available to the public as beneficial functional foods that purport to promote specific health benefits to consumers (2,14,18). In some countries probiotics are available for oral bacteriotherapy and bacterioprophylaxis of gastrointestinal disorders in humans. Often these disorders, many of which lead to diarrhea, are a direct result of antibiotic use, which produces an imbalance in the composition of the normal intestinal microbial flora. In the livestock industry the use of probiotics has potential as an alternative to antibiotics by competitive exclusion of pathogenic microorganisms (19), with some commercial products, such as Paciflor, already available. Bacteria most commonly used as probiotics include the lactic acid bacteria (e.g., lactobacilli, enterococci, streptococci, and bifidobacteria). Experimental evidence now suggests that the ingestion of substantial numbers of harmless bacteria does indeed provide a beneficial effect to the enteric flora (18). Precisely how this is achieved and whether the commercial claims are justified remains a contentious issue, though (14).In addition to the lactic acid bacteria, Bacillus species are also sold as probiotics. These consist of preparations of bacterial spores, with the potential advantage that the spore can survive transit through the stomach intact. Bacillus species are substantially different from other probiotic bacteria, though, being primarily aerobic saprophytes found in the soil. If indeed they have any health benefit, then one obviously important question is how? Do spores germinate and colonize the gut, do they competively exclude colonization by potential pathogens, or does the dormant spore provide some unique stimulus to the gut microbiota, such as enhanced local immunity?In an earlier study we have shown that one major Bacillus probiotic marketed in Europe contained spores of a taxonomically and phylogenetically unrelated Bacillus species (4). This was surprising, considering that in Europe probiotics must be licensed to be used as a functional or novel food.In this work we have examined and characterized five commercial Bacillus spore probiotics as a first step in understanding the nature of spore probiotics. MATERIALS AND METHODSBacterial strains. Bacteria were recovered by suspension of dried probiotic preparations in dist...
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