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. 2015 Mar 19;3(1):80-93.
doi: 10.3390/microorganisms3010080.

Isolation and Taxonomic Identity of Bacteriocin-Producing Lactic Acid Bacteria From Retail Foods and Animal Sources

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Free PMC article

Isolation and Taxonomic Identity of Bacteriocin-Producing Lactic Acid Bacteria From Retail Foods and Animal Sources

Chris Henning et al. Microorganisms. .
Free PMC article

Abstract

Bacteriocin-producing (Bac⁺) lactic acid bacteria (LAB) were isolated from a variety of food products and animal sources. Samples were enriched in de Man, Rogosa, and Sharpe (MRS) Lactocilli broth and plated onto MRS agar plates using a "sandwich overlay" technique. Inhibitory activity was detected by the "deferred antagonism" indicator overlay method using Listeria monocytogenes as the primary indicator organism. Antimicrobial activity against L. monocytogenes was detected by 41 isolates obtained from 23 of 170 food samples (14%) and 11 of 110 samples from animal sources (10%) tested. Isolated Bac⁺ LAB included Lactococcus lactis, Lactobacillus curvatus, Carnobacterium maltaromaticum, Leuconostoc mesenteroides, and Pediococcus acidilactici, as well as Enterococcus faecium, Enterococcus faecalis, Enterococcus hirae, and Enterococcus thailandicus. In addition to these, two Gram-negative bacteria were isolated (Serratia plymuthica, and Serratia ficaria) that demonstrated inhibitory activity against L. monocytogenes, Staphylococcus aureus, and Enterococcus faecalis (S. ficaria additionally showed activity against Salmonella Typhimurium). These data continue to demonstrate that despite more than a decade of antimicrobial interventions on meats and produce, a wide variety of food products still contain Bac⁺ microbiota that are likely eaten by consumers and may have application as natural food preservatives.

Keywords: Listeria monocytogenes; bacteriocin; food preservative; lactic acid bacteria.

Figures

Figure 1
Figure 1
Various protocols used in this study. Panel A, colony overlay assay (deferred antagonism) to identify Bac+ colonies by the sandwich-overlay technique. Panel B, isolation of Bac+ colonies from sandwich overlay plates. Panel C, the double patch plate method for purification and confirmation of Bac+ colonies and generation of the stock culture.
Figure 2
Figure 2
Bacteriocin inhibition zones from overlaid producer colonies, deferred antagonism spots, or patch-plate isolations. Panel A, representative bacteriocin inhibition zones obtained with “sandwich overlay” plates of food enrichment samples overlaid with Listeria monocytogenes 39-2 indicator lawns. Panel B, deferred antagonism assay of culture spots overlaid with Staphylococcus aureus ATCC 12600. Panel C, patch plate of Serratia plymuthica POT-1 (left) overlaid with S. aureus ATCC 12600 and an inhibition zone from Serratia ficaria CCEL-1 (right) against Salmonella Typhimurium H3380 used as a primary indicator screen.
Figure 3
Figure 3
Phylogenetic tree of 16S rRNA partial sequences from Bac+ bacteria by the Maximum Likelihood method. The phylogenetic relationship was inferred using the Tamura-Nei model [21] showing the tree with the highest log likelihood (−2162.4232). The percentage of trees in which the associated taxa clustered together is shown next to the branches.

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