Bioprotective Lactic Acid Bacteria and Lactic Acid as a Sustainable Strategy to Combat Escherichia coli O157:H7 in Meat.

Human infection by Enterohemorrhagic Escherichia coli (EHEC) constitutes a serious threat to public health and a major concern for the meat industry. Presently, consumers require safer/healthier foods with minimal chemical additives, highlighting the need for sustainable solutions to limit and prevent risks. This work evaluated the ability of two antagonistic lactic acid bacteria (LAB) strains, Lactiplantibacillus plantarum CRL681 and Enterococcus mundtii CRL35, and their combination in order to inhibit EHEC in beef (ground and vacuum sealed meat discs) at 8 °C during 72 h. The effect of lower lactic acid (LA) concentrations was evaluated. Meat color was studied along with how LAB strains interfere with the adhesion of Escherichia coli to meat. The results indicated a bacteriostatic effect on EHEC cells when mixed LAB strains were inoculated. However, a bactericidal action due to a synergism between 0.6% LA and LAB occurred, producing undetectable pathogenic cells at 72 h. Color parameters (a*, b* and L*) did not vary in bioprotected meat discs, but they were significantly modified in ground meat after 24 h. In addition, LAB strains hindered EHEC adhesion to meat. The use of both LAB strains plus 0.6% LA, represents a novel, effective and ecofriendly strategy to inactivate EHEC in meat.

Biotechnological use of dairy by-products for the production and microencapsulation of the food preservative enterocin CRL35.

Bacteriocins from Gram-positive bacteria have been proposed as natural food preservative and there is a need for large-scale production for commercial purposes. The aim of the present work is to evaluate whey, a cheese industrial by-product, for the production and microencapsulation of enterocin CRL35. Whey proved to be a promising basal medium for bacterial growth although the bacteriocin production was quite low. However, it could be much favored with the addition of yeast extract at concentrations as low as 0.5%. Besides improving bacteriocin production, this peptide was successfully microencapsulated by spray drying using whey protein concentrate and a chitosan derivative as wall materials. Microcapsules averaging 10 ± 5 µm diameter were obtained, with good structural integrity and high antimicrobial activity with a stability of at least 12 weeks at 4°C. In summary, sustainable bacteriocin production and microencapsulation was achieved recycling whey or its derivatives. In addition, the formulation owns high antimicrobial activity with a long shelf life. The development of a food preservative may represent a green solution for handling whey.

Lactiplantibacillus plantarum Strains Modulate Intestinal Innate Immune Response and Increase Resistance to Enterotoxigenic Escherichia coli Infection.

Currently, probiotic bacteria with not transferable antibiotic resistance represent a sustainable strategy for the treatment and prevention of enterotoxigenic Escherichia coli (ETEC) in farm animals. Lactiplantibacillus plantarum is among the most versatile species used in the food industry, either as starter cultures or probiotics. In the present work, the immunobiotic potential of L. plantarum CRL681 and CRL1506 was studied to evaluate their capability to improve the resistance to ETEC infection. In vitro studies using porcine intestinal epithelial (PIE) cells and in vivo experiments in mice were undertaken. Expression analysis indicated that both strains were able to trigger IL-6 and IL-8 expression in PIE cells in steady-state conditions. Furthermore, mice orally treated with these strains had significantly improved levels of IFN-γ and TNF-α in the intestine as well as enhanced activity of peritoneal macrophages. The ability of CRL681 and CRL1506 to beneficially modulate intestinal immunity was further evidenced in ETEC-challenge experiments. In vitro, the CRL1506 and CRL681 strains modulated the expression of inflammatory cytokines (IL-6) and chemokines (IL-8, CCL2, CXCL5 and CXCL9) in ETEC-stimulated PIE cells. In vivo experiments demonstrated the ability of both strains to beneficially regulate the immune response against this pathogen. Moreover, the oral treatment of mice with lactic acid bacteria (LAB) strains significantly reduced ETEC counts in jejunum and ileum and prevented the spread of the pathogen to the spleen and liver. Additionally, LAB treated-mice had improved levels of intestinal IL-10 both at steady state and after the challenge with ETEC. The protective effect against ETEC infection was not observed for the non-immunomodulatory TL2677 strain. Furthermore, the study showed that L. plantarum CRL1506 was more efficient than the CRL681 strain to modulate mucosal immunity highlighting the strain specific character of this probiotic activity. Our results suggest that the improved intestinal epithelial defenses and innate immunity induced by L. plantarum CRL1506 and CRL681 would increase the clearance of ETEC and at the same time, protect the host against detrimental inflammation. These constitute valuable features for future probiotic products able to improve the resistance to ETEC infection.