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Strain-level diversity of commercial probiotic isolates of Bacillus, Lactobacillus, and Saccharomyces species illustrated by molecular identification and phenotypic profiling.

ABSTRACT: Probiotic products are becoming more prevalent as awareness of the role of beneficial microbes in health increases. Ingredient labels of these products often omit identifications at the strain level, making it difficult to track down applicable published research. In this study, we investigated whether products labeled with the same species name contained different strains of those species. From 21 commercially available probiotic supplements and beverages, we cultured five main species: Bacillus coagulans, Bacillus subtilis, Lactobacillus plantarum, Lactobacillus rhamnosus, and the yeast Saccharomyces boulardii. To confirm the identity of each bacterial isolate, we applied standard molecular approaches: 16S rRNA gene sequencing and Matrix Assisted Laser Desorption Ionization Time-of-Flight mass spectrometry (MALDI-TOF MS). Phenotypic profiling and identification were performed with the Biolog Microbial Identification system. All of the bacterial isolates were correctly identified by at least one approach. Sequencing the 16S rRNA gene led to 82% of species identifications matching the product label, with 71% of isolates identified by MALDI-TOF MS and 60% identified correctly with the Biolog system. Analysis of the Biolog phenotypic profiles revealed different patterns of carbon source usage by each species, with sugars preferentially utilized by all except B. subtilis. To assess the strain-level differences, we compared strains of the same species and found variability in carbohydrate utilization and tolerance to environmental stressors (salt, acidity, antibiotics). By demonstrating that products listing the same species often contain strains with different 16S sequences and phenotypes, this study highlights that current labels of probiotic supplements do not sufficiently convey the strain diversity in these products.

SUBMITTER: Ansari JM

PROVIDER: S-EPMC6430388 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Project description:BACKGROUND:In the past decade, the use of probiotic-containing products has been explored as a potential alternative in oral health therapy. A widely available probiotic drink, Yakult, was evaluated for oral health applications in this longitudinal study. Selected oral health parameters, such as levels and composition of salivary and tongue plaque microbiota and of malodorous gases, in dentate healthy individuals were investigated for changes. The persistence of the probiotic strain in the oral cavity was monitored throughout the study period. METHODS:A three-phase study (7 weeks) was designed to investigate simultaneously the effect of 4-week consumption of the probiotic-containing milk drink Yakult on the microbiota of saliva and dorsum tongue coating in healthy dentate people (n = 22) and levels of volatile sulphur compounds (VSCs) in morning breath. Study phases comprised one baseline visit, at which 'control' levels of oral parameters were obtained prior to the probiotic product consumption; a 4-week period of daily consumption of one 65 ml bottle of Yakult, each bottle containing a minimum of 6.5×10(9) viable cells of Lactobacillus casei strain Shirota (LcS); and a 2-week washout period. The microbial viability and composition of saliva and tongue dorsum coating were assessed using a range of solid media. The presence of LcS in the oral cavity was investigated using a novel selective medium, 'LcS Select'. Portable sulphur monitors Halimeter(®) and OralChroma(TM) were used to measure levels of VSCs in morning breath. RESULTS:Utilization of the LcS Select medium revealed a significant (p < 0.05) but temporary and consumption-dependent presence of LcS in saliva and tongue plaque samples from healthy dentate individuals (n = 19) during the probiotic intervention phase. LcS was undetectable with culture after 2 weeks of ceasing its consumption. Morning breath scores measured with Halimeter and OralChroma were not significantly affected throughout the trial, except in a small number of individual cases where Halimeter scores were significantly reduced during the probiotic intervention period. Natural fluctuations in resident acidogenic populations, and numbers of Candida and anaerobic species, including malodourous Gram-negative anaerobes, were unaffected. CONCLUSION:While no broad ecological changes in the mouth were induced by consumption of Yakult in healthy dentate individuals, findings of this study confirm the temporary and intake-dependent presence of LcS. Future studies could focus on subjects at greater risk of oral infection, where ill-defined microbiota (e.g. an increased presence of periopathogens) or clinically diagnosed halitosis might be significantly affected by consumption of this probiotic.

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Strain-level diversity of commercial probiotic isolates of Bacillus, Lactobacillus, and Saccharomyces species illustrated by molecular identification and phenotypic profiling.

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