title: Feasibility of Genome-Wide Screening for Biosafety Assessment of Probiotics: A Case Study of Lactobacillus helveticus MTCC 5463.
abstract: Recent years have witnessed an explosion in genome sequencing of probiotic strains for accurate identification and characterization. Regulatory bodies are emphasizing on the need for performing phase I safety studies for probiotics. The main hypothesis of this study was to explore the feasibility of using genome databases for safety screening of strains. In this study, we attempted to develop a framework for the safety assessment of a potential probiotic strain, Lactobacillus helveticus MTCC 5463 based on genome mining for genes associated with antibiotic resistance, production of harmful metabolites, and virulence. The sequencing of MTCC 5463 was performed using GS-FLX Titanium reagents. Genes coding for antibiotic resistance and virulence were identified using Antibiotic Resistance Genes Database and Virulence Factors Database. Results indicated that MTCC 5463 carried antibiotic resistance genes associated with beta-lactam and fluoroquinolone. There is no threat of transfer of these genes to host gut commensals because the genes are not plasmid encoded. The presence of genes for adhesion, biofilm, surface proteins, and stress-related proteins provides robustness to the strain. The presence of hemolysin gene in the genome revealed a theoretical risk of virulence. The results of in silico analysis complemented the in vitro studies and human clinical trials, confirming the safety of the probiotic strain. We propose that the safety assessment of probiotic strains administered live at high doses using a genome-wide screening could be an effective and time-saving tool for identifying prognostic biomarkers of biosafety.
authors.affiliation: Department of Dairy Science, South Dakota State University, Brookings, SD, 57007, USA. Suja.Senan@sdstate.edu.
authors.forename: S
authors.initials: S
authors.lastname: Senan
authors.affiliation: Department of Dairy Microbiology, Anand Agricultural University, Anand, 388110, India.
authors.forename: J B
authors.initials: JB
authors.lastname: Prajapati
authors.affiliation: Department of Animal Biotechnology, Anand Agricultural University, Anand, 388110, India.
authors.forename: C G
authors.initials: CG
authors.lastname: Joshi
mesh_terms: D003257:Consumer Product Safety; D030541:Databases, Genetic; D024881:Drug Resistance, Bacterial; D005240:Feasibility Studies; D016680:Genome, Bacterial; D006460:Hemolysin Proteins / Q000235:genetics; D052198:Lactobacillus helveticus / Q000187:drug effects / Q000235:genetics / Q000472:pathogenicity; D019936:Probiotics; D014774:Virulence / Q000235:genetics
chemical_list: D006460:Hemolysin Proteins
keywords: 16S rRNA; Antibiotic resistance; Lactobacillus spp.; Probiotics