Genome-based taxonomic identification and safety assessment of an Enterococcus strain isolated from a homemade dairy product.

The aim of this study was to explore the taxonomic identification and evaluate the safety of a bacterium, Enterococcus lactis IDCC 2105, isolated from homemade cheese in Korea, using whole genome sequence (WGS) analysis. It sought to identify the species level of this Enterococcus spp., assess its antibiotic resistance, and evaluate its virulence potential. WGS analysis confirmed the bacterial strain IDCC 2105 as E. lactis and identified genes responsible for resistance to erythromycin and clindamycin, specifically msrC, and eatAv, which are chromosomally located, indicating a minimal risk for horizontal gene transfer. The absence of plasmids in E. lactis IDCC 2105 further diminishes the likelihood of resistance gene dissemination. Additionally, our investigation into seven virulence factors, including hemolysis, platelet aggregation, biofilm formation, hyaluronidase, gelatinase, ammonia production, and ß-glucuronidase activity, revealed no detectable virulence traits. Although bioinformatic analysis suggested the presence of collagen adhesion genes acm and scm, these were not corroborated by phenotypic virulence assays. Based on these findings, E. lactis IDCC 2105 presents as a safe strain for potential applications, contributing valuable information on its taxonomy, antibiotic resistance profile, and lack of virulence factors, supporting its use in food products.

Isolation, Identification, and Characterization of the Native Yeast Strains from Homemade Cheese to Assess their Eliminating Impact on the Aflatoxin B1 and M1 of the Simulated Gastrointestinal Fluid.

Background: The occurrence of aflatoxins in food products is a silent threat to human health worldwide. A range of strategies has been introduced to address the bioavailability of aflatoxins, which are considered microbial tools to provide a low-cost and promising approach. Objectives: The present study focused on the separation of yeast strains from the homemade cheese rind layer to investigate the ability of native yeasts to eliminate AB1 and AM1 from simulated gastrointestinal fluids. Material and Methods: Homemade cheese samples were prepared from different locations in Tehran provinces and yeast strains were isolated and identified through the biochemical methods and molecular analysis of internal transcribed spacer and D1/D2 domain of 26S rDNA regions. Isolated strains were screened using simulated gastrointestinal fluids, and the ability of yeast strains to absorb aflatoxin was evaluated. Results: Out of 13 strains, 7 yeast strains were not affected by 5 ppm AFM1 while 11 strains did not show any significant response to 5 mg.L-1 (ppm) of AFB1. On the other hand, 5 strains were able to successfully tolerate 20 ppm AFB1. Candidate yeasts showed different abilities to remove aflatoxins B1 and M1. In addition, C. lusitaniae, G. geotrichum, G. candidum, and C. sanyaensis exhibited a significant ability to detoxify aflatoxins from the gastrointestinal fluid, respectively. Conclusion: Our data suggest that yeast communities with essential effects on the quality of homemade cheese appear to be precise candidates for the potential elimination of aflatoxins from the gastrointestinal fluid.

Safety evaluation of Lactococcus lactis IDCC 2301 isolated from homemade cheese.

For applications of microorganisms as probiotics in the food industry, safety evaluation has increasingly become important to ensure the health of consumers. Although people have been using various lactic acid bacteria for different purposes, some studies have reported that certain lactic acid bacteria exhibit properties of virulence and produce toxic compounds. Thus, it is necessary to examine the characteristics associated with lactic acid bacteria that are safe for use as probiotics. This research aimed to assess the safety of Lactococcus lactis IDCC 2301 isolated from homemade cheese using in vitro and in vivo assays, including antibiotic resistance, hemolytic activity, toxin production, infectivity, and metabolic activity in immune-compromised animal species. The results demonstrated that the strain was susceptible to nine antibiotics suggested by the European Food Safety Authority (EFSA). Whole-genome analysis revealed that L. lactis IDCC 2301 neither has toxigenic genes nor harbors antibiotic resistance. Moreover, L. lactis IDCC 2301 showed neither hemolytic nor ß-glucuronidase activity. Furthermore, none of the D-lactate and biogenic amines were produced by L. lactis IDCC 2301. Finally, it was demonstrated that there was no toxicity and mortality using single-dose oral toxicity tests in rats. These results indicate that L. lactis IDCC 2301 can be safely used as probiotics for human consumption.

Occurrence of non-O157 Shiga toxin-encoding Escherichia coli in artisanal mozzarella cheese in Brazil: risk factor associated with food workers

Abstract Mozzarella cheese is considered a safe food due to the high temperature treatment used in the traditional process of stretching into hot water; however, a post-process contamination during the cheese manufacture or during the processing (before distribution) could occur. This study investigated the occurrence of Shiga toxin-encoding Escherichia coli (STEC) strains in homemade-mozzarella cheese. In total, 59 raw milk cheese samples collected at a local producer in the Jequitinhonha Valley (Northeast of Minas Gerais, Brazil) were submitted for microbiological analysis, and 38 (64.4%) tested positive for E. coli. Then, 147 strains of E. coli were isolated from positive samples and screened by polymerase chain reaction for the presence of the genes encoding the following virulence factors: stx1 and stx 2 (verotoxin types 1 and 2) and eae (intimin). All the isolates were negative for the stx 2, 14 isolates (9.5%) were positive for the stx1 gene, and 11 of them also harbored the eae gene. A food worker was probably involved in cheese contamination during the manufacture schedule. While the development of STEC infection in humans is not completely understood, all STEC- contaminated food can be considered potentially hazardous.