Lactobacillus reuteri and Enterococcus faecium from Poultry Gut Reduce Mucin Adhesion and Biofilm Formation of Cephalosporin and Fluoroquinolone-Resistant Salmonella enterica

Abstract

Non-typhoidal Salmonella (NTS) can cause infection in poultry, livestock, and humans. Although the use of antimicrobials as feed additives is prohibited, the previous indiscriminate use and poor regulatory oversight in some parts of the world have resulted in increased bacterial resistance to antimicrobials, including cephalosporins and fluoroquinolones, which are among the limited treatment options available against NTS. This study aimed to isolate potential probiotic lactic acid bacteria (LAB) strains from the poultry gut to inhibit fluoroquinolone and cephalosporin resistant MDR Salmonella Typhimurium and S. Enteritidis. The safety profile of the LAB isolates was evaluated for the hemolytic activity, DNase activity, and antibiotic resistance. Based on the safety results, three possible probiotic LAB candidates for in vitro Salmonella control were chosen. Candidate LAB isolates were identified by 16S rDNA sequencing as Lactobacillus reuteri PFS4, Enterococcus faecium PFS13, and Enterococcus faecium PFS14. These strains demonstrated a good tolerance to gastrointestinal-related stresses, including gastric acid, bile, lysozyme, and phenol. In addition, the isolates that were able to auto aggregate had the ability to co-aggregate with MDR S. Typhimurium and S. Enteritidis. Furthermore, LAB strains competitively reduced the adhesion of pathogens to porcine mucin Type III in co-culture studies. The probiotic combination of the selected LAB isolates inhibited the biofilm formation of S. Typhimurium FML15 and S. Enteritidis FML18 by 90% and 92%, respectively. In addition, the cell-free supernatant (CFS) of the LAB culture significantly reduced the growth of Salmonella in vitro. Thus, L. reuteri PFS4, E. faecium PFS13, and E. faecium PFS 14 are potential probiotics that could be used to control MDR S. Typhimurium and S. Enteritidis in poultry. Future investigations are required to elucidate the in vivo potential of these probiotic candidates as Salmonella control agents in poultry and animal feed.

Evaluating Enterococcus faecium9 N‐2 as a probiotic candidate from traditional village white cheese

“…There was a 21% increase in viability compared to the initial concentration. Similar results for E. faecium were observed at different phenol concentrations (0.25%, 0.40%, and 0.50%) for 18-24 h at 30-37°C (Sakkaa et al, 2022;Siddique et al, 2021;Tilwani et al, 2022;Vadassery & Pillai, 2020).…”

Anti-Quorum Sensing Activity of Probiotics: The Mechanism and Role in Food and Gut Health

“…acidophilus A4 and CRL 730, respectively [ 25 , 29 ]. The anti-biofilm and anti-QS activity of probiotics against Salmonella was also observed, but the QSIs were not identified [ 35 , 36 , 37 , 38 , 39 ]. In these studies, EPS might be produced by the lactobacillus species as the QSI.…”

“…coryniformis NA-3. Even though not investigated, EPS from the Lactobacillus species could play a role in the interference with the QS in Salmonella through the downregulation of the genes encoding the QS regulator proteins [ 35 , 36 , 38 , 39 , 40 ]. Unfortunately, the mechanisms behind the QS inhibition against E. coli and Salmonella were not well identified.…”

Recombinant Limosilactobacillus (Lactobacillus) delivering nanobodies against Clostridium perfringens NetB and alpha toxin confers potential protection from necrotic enteritis

“…Many Limosilactobacillus species have “Qualified Presumption of Safety (QPS)” status from the European Food Safety Authority (EFSA) and “Generally Regarded As Safe (GRAS)” status from US Food and Drug Administration (FDA). Lactobacilli are key members of the endogenous microbiota of oral, GI, respiratory, and urogenital mucosa and play a key role in regulating local microbiota, restoring barrier function, preventing inflammation associated with GI diseases, improving growth performance, and protecting against infectious diseases (Chen et al, 2021 ; Han et al, 2021 ; Hu et al, 2021 ; Klaenhammer et al, 2002 ; Li et al, 2018 ; Manes‐Lazaro et al, 2017 ; Ouwehand et al, 2002 ; Siddique et al, 2021 ; Vineetha et al, 2017 ; Y. Wang et al, 2021 ; Wu et al, 2021 ). Of particular importance, Lactobacilli have been shown to possess strong antagonistic activity against C. perfringens , inhibit toxin production, reduce proinflammatory cytokines, improve intestinal integrity and immune response, correct microbial dysbiosis, restore performance deficiencies associated with subclinical NE, and protect chickens from clinical NE (Dec et al, 2016 ; Gong et al, 2020 ; Guo et al, 2017 , 2021 ; Khalique et al, 2019 ; Kizerwetter‐Swida & Binek, 2005 , 2009 , 2016 ; La Cao et al, 2012 ; La Ragione et al, 2004 ; Li et al, 2017 ; Shojadoost et al, 2022 ; Xu et al, 2020 ).…”

“…Many Limosilactobacillus species have "Qualified Presumption of Safety (QPS)" status from the European Food Safety Authority (EFSA) and "Generally Regarded As Safe (GRAS)" status from US Food and Drug Administration (FDA). Lactobacilli are key members of the endogenous microbiota of oral, GI, respiratory, and urogenital mucosa and play a key role in regulating local microbiota, restoring barrier function, preventing inflammation associated with GI diseases, improving growth performance, and protecting against infectious diseases (Chen et al, 2021;Han et al, 2021;Hu et al, 2021;Klaenhammer et al, 2002;Li et al, 2018;Manes-Lazaro et al, 2017;Ouwehand et al, 2002;Siddique et al, 2021;Vineetha et al, 2017; Y. Wu et al, 2021).…”