Salmonella enterica serovar Typhimurium (ST) remains a major infectious agent in the USA, with an increasing antibiotic resistance pattern, which requires the development of novel antimicrobials capable of controlling ST. Polyphenolic compounds found in plant extracts are strong candidates as alternative antimicrobials, particularly phenolic acids such as gallic acid (GA), protocatechuic acid (PA) and vanillic acid (VA). This study evaluates the effectiveness of these compounds in inhibiting ST growth while determining changes to the outer membrane through fluorescent dye uptake and scanning electron microscopy (SEM), in addition to measuring alterations to virulence genes with qRT-PCR. Results showed antimicrobial potential for all compounds, significantly inhibiting the detectable growth of ST. Fluorescent spectrophotometry and microscopy detected an increase in relative fluorescent intensity (RFI) and red-colored bacteria over time, suggesting membrane permeabilization. SEM revealed severe morphological defects at the polar ends of bacteria treated with GA and PA, while VA-treated bacteria were found to be mid-division. Relative gene expression showed significant downregulation in master regulator hilA and invH after GA and PA treatments, while fliC was upregulated in VA. Results suggest that GA, PA and VA have antimicrobial potential that warrants further research into their mechanism of action and the interactions that lead to ST death.

In this study, the effect of sustainable probiotics on Campylobacter jejuni colonization and gut microbiome composition was evaluated using chicken as a model organism. Chickens were given Lactobacillus casei over-expressing myosin-cross-reactive antigen (LC+mcra). LC+mcra can generate bioactive compounds in larger quantity including conjugated linoleic acid. A total of 120 chickens were used in duplicate trials to investigate the effectiveness of LC+mcra in decreasing C. jejuni colonization by means of kanamycin resistant strain compared to the control group. We observed that LC+mcra can efficiently colonize various parts of the chicken gut and competitively reduce colonization of natural and challenged C. jejuni and natural Salmonella enterica. LC+mcra was found to reduce C. jejuni colonization in cecum, ileum and jejunum, by more than one log CFU/g when compared to the no-probiotic control group. Furthermore, 16S rRNA compositional analysis revealed lower abundance of Proteobacteria, higher abundance of Firmicutes, along with enriched bacterial genus diversity in gut of LC+mcra fed chicken. Decreased contamination of drinking water by C. jejuni and S. enterica was also observed, suggesting a potential function of reducing horizontal transfer of enteric bacteria in poultry. Outcomes of this study reveal high potential of LC+mcra as sustainable approach to decrease colonization of C. jejuni and S. enterica in poultry gut along with other beneficial attributes.

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Probiotics in fermented foods or commercially available supplements benefit the host by providing metabolites and peptides. The production of these metabolites varies with the available substrates or prebiotics present in the system and their concentration. In this study, 0.5% peanut flour (PF) was used to stimulate the growth and production of metabolites of wild-type Lactobacillus casei (LCwt) and compare with an engineered L. casei (LCCLA) capable of converting a higher amount of conjugated linoleic acid (CLA). The total extracellular metabolites present in the cell-free cultural supernatant (CFCS) of LCwt (without peanut), LCwt+PF (with peanut), and LCCLA were collected after 24 and 48 h of incubation, and their antagonistic activities against enterohemorrhagic Escherichia coli (EHEC EDL933) growth and pathogenesis were evaluated. All collected metabolites exhibited varying efficiency in restraining EHEC EDL933 growth, whereas supplementing a low concentration of CLA to the 48-h CFCS from LCwt showed augmented antagonism toward EHEC EDL933. A downregulation of key virulence genes was observed from metabolites collected at the 48-h time point. These observations indicate that the presence of metabolites in CFCSs—including CLA, which is produced by Lactobacillus and was identified by gas chromatography–mass spectrometry—plays a critical role. This study demonstrates the potential applicability of Lactobacillus-originated CLA in the prevention of EHEC EDL933–mediated illnesses. HIGHLIGHTS

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