Abstract
Antimicrobial potential of a dairy-origin probiotic bacteria, Propionibacterium freudenreichii, against multidrug-resistant Salmonella Heidelberg (SH) in turkey poults was determined in the current study. Employing in vitro experiments, two strains (subsp.) of P. freudenreichii: P. freudenreichii freudenreichii B3523 (PF) and P. freudenreichii shermanii B4327 (PS) were tested for their ability to resist low pH (2.5) and bile salts (0.3%). In addition, the ability of the strains to adhere to and invade avian epithelial cells was determined after exposure to Propionibacterium strains followed by SH challenge. Moreover, the antibacterial activity of the strains’ cell-free culture supernatants (CFCSs) were tested against three major foodborne pathogens, including SH. Furthermore, the susceptibility of the strains to common antibiotics used for human therapy was determined. The hemolytic properties of the strains were determined in comparison to Streptococcus pyogenes, a known hemolysis-causing pathogen. Appropriate controls were kept in all studies. Using two in vivo experiments, PF was tested against SH colonization of poult ceca and dissemination to liver and spleen. The four treatment groups were: negative control, PF control (PFC), SH control (SC), and a test group (PFS; PF + SH). The poults in the PFC and PFS groups were inoculated with 1010 CFU ml−1 PF on day 1 through crop gavage and subsequently supplemented through drinking water. On day 7, SC and PFS groups were challenged with SH at 106 CFU ml−1, and after 7 days, cecum, liver, and spleen were collected for determining surviving SH populations. Results indicated that both PF and PS resisted pH = 2.5 and 0.3% bile salts with surviving populations comparable to the control and adhered well onto the avian epithelial cell lines. The strains were susceptible to antibiotics and did not invade the epithelial cells or exhibit hemolytic properties. The CFCSs were highly bactericidal against all tested pathogens. In turkey poults, PF significantly reduced cecal colonization of SH and the dissemination of the pathogen to the liver, compared to the SH challenge controls (P < 0.05). Results revealed that PF, a non-host gastrointestinal tract-derived probiotic, could be an antibiotic alternative to prevent the early colonization of SH in poults, improving the preharvest safety of turkeys.
Highlights
Non-typhoidal Salmonella is a major bacterial pathogen that causes ∼11% illnesses, 35% hospitalizations, and 28% deaths associated with foodborne outbreaks in the United States, annually (Scallan et al, 2011)
The results of the current study revealed that both P. freudenreichii strains were highly tolerant to the low pH used in the study
We found that P. freudenreichii subsp. freudenreichii (PF) and P. freudenreichii subsp. shermanii (PS) exhibited high survivability with viable populations of 8.17- and 8.04- log10 CFU ml−1, respectively, in the treatments when exposed to 0.3% bile salts for 3.5 h, comparable to the controls (Figures 2A,B)
Summary
Non-typhoidal Salmonella is a major bacterial pathogen that causes ∼11% illnesses, 35% hospitalizations, and 28% deaths associated with foodborne outbreaks in the United States, annually (Scallan et al, 2011). The occurrence of MDR SH in turkeys is a serious concern since it is a major poultry species produced and consumed in the United States and exported globally in massive volumes (USPEA, 2017) This situation is aggravated due to the increased invasiveness of SH isolated from poultry and poultry products, and their multiple drug resistance profiles (Foley et al, 2011; Medeiros et al, 2011; Hoffmann et al, 2014; Antunes et al, 2016). This step has resulted in the rigorous search for antibiotic alternatives that can produce meaningful reductions of pathogenic bacteria in food animals, thereby reducing the risk of contaminated animal products entering the food chain
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