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Use of probiotic biofilms can be an alternative approach for reducing the formation of pathogenic biofilms in food industries. The aims of this study were (i) to evaluate the probiotic properties of bacteriocinogenic ( Lactococcus lactis  VB69,  L. lactis  VB94,  Lactobacillus sakei  MBSa1, and  Lactobacillus curvatus  MBSa3) and non-bacteriocinogenic ( L. lactis  368,  Lactobacillus helveticus  354,  Lactobacillus casei  40, and  Weissela viridescens  113) lactic acid bacteria (LAB) isolated from Brazilian’s foods and (ii) to develop protective biofilms with these strains and test them for exclusion of  Listeria monocytogenes, Escherichia coli  O157:H7, and  Salmonella  Typhimurium. LAB were tested for survival in acid and bile salt conditions, surface properties, biosurfactant production, β-galactosidase and gelatinase activity, antibiotic resistance and presence of virulence genes. Most strains survived exposure to pH 2 and 4% bile salts. The highest percentages of auto-aggregation were obtained after 24 h of incubation. Sixty-seven percentage auto-aggregation value was observed in  W. viridescens  113 and  Lactobacillus curvatus  MBSa3 exhibited the highest co-aggregation (69% with  Listeria monocytogenes  and 74.6% with  E. coli  O157:H7), while the lowest co-aggregation was exhibited by  W. viridescens  113 (53.4% with  Listeria monocytogenes  and 38% with  E. coli  O157:H7). Tests for hemolytic activity, bacterial cell adherence with xylene, and drop collapse confirmed the biosurfactant-producing ability of most strains. Only one strain ( L. lactis  368) produced β-galactosidase. All strains were negative for virulence genes  cob, ccf, cylLL, cylLs, cyllM, cylB, cylA  and  efaAfs  and gelatinase production. The antibiotic susceptibility tests indicated that the MIC for ciprofloxacin, clindamycin, gentamicin, kanamycin, and streptomycin did not exceed the epidemiological cut-off suggested by the European Food Safety Authority. Some strains were resistant to one or more antibiotics and resistance to antibiotics was species and strain dependent. In the protective biofilm assays, strains  L. lactis  368 (bac-),  Lactobacillus curvatus  MBSa3 (bac+), and  Lactobacillus sakei  MBSa1 (bac+) resulted in more than six log reductions in the pathogens counts when compared to the controls. This effect could not be attributed to bacteriocin production. These results suggest that these potential probiotic strains can be used as alternatives for control of biofilm formation by pathogenic bacteria in the food industry, without conferring a risk to the consumers.

frontiers in microbiology

Use of Potential Probiotic Lactic Acid Bacteria (LAB) Biofilms for the Control of Listeria monocytogenes, Salmonella Typhimurium, and Escherichia coli O157:H7 Biofilms Formation