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. 2018 Aug 6:9:1685.
doi: 10.3389/fmicb.2018.01685. eCollection 2018.

Evaluation of Probiotic Properties and Safety of Enterococcus faecium Isolated From Artisanal Tunisian Meat "Dried Ossban"

Mohamed Zommiti  1 , Mélyssa Cambronel  2 , Olivier Maillot  2 , Magalie Barreau  2 , Khaled Sebei  1 , Marc Feuilloley  2 , Mounir Ferchichi  1   3 , Nathalie Connil  2
Affiliations

Evaluation of Probiotic Properties and Safety of Enterococcus faecium Isolated From Artisanal Tunisian Meat "Dried Ossban"

Mohamed Zommiti et al. Front Microbiol. .

Abstract

Enterococcus faecium strains were isolated from an original biotope, artisanal dried Tunisian meat "Dried Ossban," and evaluated for safety and capacity as probiotics. Gram-positive, catalase negative, and bacteriocin-producing bacteria were screened using selective microbiological media. All isolates were identified by phenotypic and molecular tools. Five E. faecium strains (MZF1, MZF2, MZF3, MZF4, and MZF5) were selected and further assessed for their probiotic properties. They were found to be resistant to the physiological concentrations of bile salts, and the harsh conditions of the gastrointestinal tract, and showed autoaggregation and adhesion ability. All these isolates possess at least one enterocin and could efficiently inhibit the growth of Listeria innocua HPB13. The analysis of their safety profile revealed for almost all the strains the absence of cytotoxicity and virulence determinants, and susceptibility to clinically important antibiotics such as vancomycin. These data suggest that these bacteria, isolated from "Dried Ossban," do not present a risk to human health, and may be considered as interesting candidates for future use as probiotics and bioprotective cultures for application in the food and/or feed industries.

Keywords: Dried Ossban; Enterococcus faecium; antibiotic resistance; probiotics; safety; virulence determinants.

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Figures

FIGURE 1
FIGURE 1
MALDI-TOF MS based phylogenetic tree. (A) Distance level between the E. faecium strains isolated from Tunisian “Dried Ossban” and (B) distance level of the isolates compared to the E. faecium reference strains available in the Biotyper library.
FIGURE 2
FIGURE 2
Autoaggregation of the E. faecium strains isolated from “Dried Ossban.” Data are expressed as mean ± SE.
FIGURE 3
FIGURE 3
Adhesion of the E. faecium isolates to the intestinal Caco-2/TC7 cells. Data are expressed as mean ± SE.
FIGURE 4
FIGURE 4
Cytotoxicity of the E. faecium isolates measured by LDH release after overnight incubation of Caco-2/TC7 cells with 108 bacteria/ml. Data are expressed as mean ± SE.
FIGURE 5
FIGURE 5
TEER at time 0, 14, and 24 h of Caco-2/TC7 cells exposed to the E. faecium isolates (108 bacteria/ml). Data are expressed as percentages of the initial level measured in the insert and as a mean ± SE. ∗P < 0.05 and ∗∗P < 0.01 compared to Caco-2/TC7 control.
FIGURE 6
FIGURE 6
PCR amplification of the tyrosine decarboxylase (tdc) gene in the five E. faecium strains isolated from “Dried Ossban” Molecular Weight marker (MW) 1 kB.
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