Pediococcus Acidilactici Inhibit Biofilm Formation of Food-Borne Pathogens on Abiotic Surfaces

In this study,we aimed to examine the inhibitory effect of PA003,a Pediococcus acidilactici that produces lactic acid and antimicrobial peptides pediocin,on pathogenic biofilm formation on abiotic surfaces.PA003 and pathogens(Escherichia coli,Salmonella enterica serovar Typhimurium,Staphylococcus aureus and Listeria monocytogenes) were used to evaluate auto-aggregation,hydrophobicity,biofilm formation and biofilm formation inhibition on stainless steel,polyvinyl chloride and glass slides in terms of exclusion,displacement and competition.The results showed the highest auto-aggregation abilities were observed for one of the E.coli strains EAggEC(E58595) and the highest hydrophobic strain was observed with EPEC(E2348/69)(51.9%).The numbers of biofilm cells of E.coli,S.Typhimurium,S.aureus and L.monocytogenes on stainless steel,polyvinyl chloride and glass slide coupons were effectively reduced by approximately 4log CFU/coupon.These results demonstrate that lactic acid bacteria can be used as an alternative to effectively control the formation of biofilms by food-borne pathogens.

Green Fluorescent Protein Recombinant Nisin as a Probe for Detection of Gram-Positive Bacteria

A great amount of foodborne pathogens were Gram-positive(G+) bacteria, a threat to public health. In this study, considering the binding ability of nisin towards G+ bacteria and the stable fluorescent ability of EGFP protein, a fluorescent nisin–EGFP protein probe was constructed by a gene engineering method. Nisin and EGFP were used as the receptor and fluorophore, respectively, to detect G+ bacteria. The nisin and egfp gene were amplified separately according to the sequence published in Gen Bank using unique primers. The two genes were cloned into a pET-28b(+) vector resulting in apET-28b(+)–nisin–egfp vector. The vector was transferred into Escherichia coli(E. coli) BL21(DE3) for expression. The expressed protein was extracted, purified by a Ni–NTA column, and then tested by the SDS-PAGE method to confirm its molecular weight. Listeria monocytogenes(L.monocytogenes), Staphylococcus aureus(S. aureus), and Micrococcus luteus(M. luteus) were used as the representations of G+ bacteria. E. coli O157, representing the gram-negative(G-) bacteria, was used as a negative control. The binding specificity of the recombinant protein was performed on two types of bacteria and then detected through fluorescent microscopy. The results indicated that the nisin–EGFP probe could detect G+ bacteria at 10~8CFU/mL.