Probiotic Activity of Enterococcus faecium and Lactococcus lactis Isolated from Thai Fermented Sausages and Their Protective Effect Against Clostridium difficile

Abstract

Lactic acid bacteria, Enterococcus faecium and Lactococcus lactis, previously isolated from Thai fermented sausages were elucidated their probiotic properties especially in the control of Clostridium difficile 630. Both isolates survived in simulated gastric solution at pH 3 followed in simulated intestinal solution at pH 8. The presence of skimmed milk also helped the bacteria to survive through acidic and alkaline in gastrointestinal conditions. The adhesion properties of both isolates were tested using a human colon adenocarcinoma cell line. The result showed that both isolates exhibited desirable probiotic properties which adhered to Caco-2 cells. The neutralized cell-free supernatant of both isolates demonstrated that no cytotoxicity toward Caco-2 cells vice versa cell-free supernatant of C. difficile 630 toward Caco-2 cell demonstrated high toxicity. The immunomodulation effect in response to bacterial neutralized cell-free supernatant and cell-free supernatant was also studied. The expression level of pro-inflammatory cytokine of Caco-2 cell which are tumor necrosis factor-α and interleukin-8 was evaluated using quantitative reverse transcriptase PCR. Both isolates were able to diminish the expression level of TNF-α and IL-8 induced by the cell-free supernatant of C. difficile 630. Hence, these isolates would be able to improve the gut health through counteracting the C. difficile-associated intestinal inflammation in human cell lines. These results may contribute to the development of the isolates using as probiotics.

Characterization of Lactic Acid Bacteria Isolated From the Gastrointestinal Tract of a Wild Boar as Potential Probiotics

“…Similar results have been observed in the current study that the L. mucosae M6-29 and L. salivarius M2-71, with strong aggregation ability, have stronger capacity to adhere to cells than other isolates. It is supposed that aggregation factors which favorably increase H-bonding between and among cell surfaces have correlation with increased general adhesion ability (29). Our results support the hypothesis that a notable correlation was observed between aggregation and adhesion.…”

“…The Caco-2 cells and IPEC-J2 cells were cultivated as described by Dowdell et al (29). Briefly, the cells were plated into 24well plates.…”

Enterococcus faecalis Isolated From Infant Feces Inhibits Toxigenic Clostridioides (Clostridium) difficile

“…They showed that Bifidobacterium longum and Bifidobacterium breve were able to reduce the toxic effects of the pathogen on HT-29 cell lines and rounding was prevented and F-actin microstructure and tight junctions between adjacent cells were preserved. E. faecium and L. lactis have also been shown to help protect epithelial cells from C. difficile toxins (11). Similarly, the results of our study showed that six E. faecalis may secrete antibacterial agents that reduce the cytotoxic effects of toxins of C. difficile and protect HT-29 cells.…”

“…Various antimicrobial mechanisms have been attributed to LAB probiotics. These include nutrient competition, prevention of mucosal cell adhesion, and internalization of pathogens such as C. difficile ( 9 , 11 , 12 ). Moreover, LAB strains can produce lactic acid and certain antimicrobial molecules such as hydrogen peroxide, fatty acid, and bacteriocins to exert their antimicrobial activity ( 13 , 14 ).…”

Postbiotics against Pathogens Commonly Involved in Pediatric Infectious Diseases

“…New evidence shows that CFS from various lactic acid bacteria (LAB) cocultured with multi-resistant C. difficile can reduce the cytotoxic effects of clostridial toxins on different cell lines compared to pure C. difficile CFS. Additionally, neutralized CFS from LAB + C. difficile can reduce the level of IL-8 and tumor necrosis factor α (TNF-α), two pro-inflammatory cytokines released by intestinal epithelial cells, caused by the CFS of C. difficile [ 67 , 68 ].…”