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Recently, the interest in donkey milk has increased considerably because it proved high nutritive and functional values of their ingredients. Its chemical composition is widely studied, but its microbiota, especially lactic acid bacteria, remains less studied. This study focuses on analyzing, isolating, and identifying lactic acid bacteria and evaluating their capacity to produce biomolecules with antibacterial activity. Among 44 strains identified, 43 are Gram-positive, and most are catalase-negative and cocci-shaped. Five strains were selected to evaluate their antibacterial activity against Listeria monocytogenes, Staphylococcus aureus, and Escherichia coli. Different induction methods allowed to amplify the antibacterial effects against these pathogenic strains.

Raman optical spectroscopy promises label-free bacterial detection, identification, and antibiotic susceptibility testing in a single step. However, achieving clinically relevant speeds and accuracies remains challenging due to weak Raman signal from bacterial cells and numerous bacterial species and phenotypes. Here we generate an extensive dataset of bacterial Raman spectra and apply deep learning approaches to accurately identify 30 common bacterial pathogens. Even on low signal-to-noise spectra, we achieve average isolate-level accuracies exceeding 82% and antibiotic treatment identification accuracies of 97.0±0.3%. We also show that this approach distinguishes between methicillin-resistant and -susceptible isolates of Staphylococcus aureus (MRSA and MSSA) with 89±0.1% accuracy. We validate our results on clinical isolates from 50 patients. Using just 10 bacterial spectra from each patient isolate, we achieve treatment identification accuracies of 99.7%. Our approach has potential for culture-free pathogen identification and antibiotic susceptibility testing, and could be readily extended for diagnostics on blood, urine, and sputum. The use of Raman spectroscopy for pathogen identification is hampered by the weak Raman signal and phenotypic diversity of bacterial cells. Here the authors generate an extensive dataset of bacterial Raman spectra and apply deep learning to identify common bacterial pathogens and predict antibiotic treatment from noisy Raman spectra.

The morphogenesis of ovococci has been reviewed extensively. Recent results have provided new insights concerning the mechanisms of elongation in ovoid bacteria. We present here the proteins involved in the elongation (firmly established and more or less hypothetical) and discuss the relationship between elongation and division of ovococci.

Gold nanoparticles (AuNPs) have become frequently used materials in biotechnological and biomedical applications including cancer. They can be commonly synthesized by biological and chemical methods. In the present study, we synthesized Enterococcus -mediated AuNPs and evaluated their cytotoxicity in human colorectal cancer cell line (HT-29). AuNPs are synthesized intracellularly using Enterococcus sp. RMAA. Characterization of AuNPs has done using UV spectrophotometry and transmission electron microscope. Cytotoxicity was evaluated by MTT assay. Intercellular reactive oxygen species (ROS) expression and apoptosis-related morphology were evaluated by dichlorodihydrofluorescein diacetate and acridine orange/ethidium bromide staining via fluorescence microscopy. JC-1 staining and caspase 3 immunofluorescence expression were analyzed by confocal microscopy. Enterococcus sp. RMAA–mediated AuNPs are spherical and induced concentration-dependent cytotoxicity in HT-29 cells. AuNP treatments also induced ROS and caspase-3 expressions and reduced the mitochondrial membrane potential. Morphology related to apoptotic changes was also noticed after AuNP treatments in HT-29 cells. The present study revealed that Enterococcus -derived AuNPs induced apoptotic cell death in HT-29 cells and suggests that AuNPs could be used as a pro apoptotic agent for colon cancer treatment.

The occurence and spread of animal-derived methicillin resistant staphylococci (MRS) worldwide is a current problem, especially due to their increasing incidence in food animals and their products, with possible contamination of food consumers and handlers. Staphylococci isolated from animals (n = 123) were identified with MALDI-TOF mass spectrometry and screened for methicillin/oxacillin/cefoxitin resistance (MR) using the disk diffusion method. Twenty-three phenotypically MRS strains were analysed using PBP2' Latex Agglutination Test Kit to confirm the phenotypic MR and PCR was performed for mecA gene detection; mecA gene positive strains were furtherly confirmed by means of sequencing. The susceptibility of MRS to 11 partially-purified enterocins (Ent) produced by E. faecium, E. durans and E. mundtii strains of animal, feed/food and environmental origin was checked using agar spot tests. Out of 23 MRS, PBP testing confirmed MR in 17 strains. Three Staphylococcus epidermidis and one S. vitulinus were mecA positive. The majority of MRS, including two mecA gene-positive strains S. epidermidis R44/1 and P3/Tr2a, were susceptible to the tested enterocins, mainly to Ent7420, EntA(P)/EK13, Ent412, Ent55 and Ent9296 (in the range 100 – 12,800 AU/mL). The most susceptible strains appeared to be the mecA gene-positive S. epidermidis SE R44/1 and SE P3/Tr2a strains, inhibited by eight enterocins out of 11 tested (100–200 AU/mL). Only four strains (including mecA gene positive S. epidermidis SE P3/Tr1 and S. vitulinus SV K12PL/1) were resistant to the tested antimicrobial substances. These results indicate that the enterocins used offer a promising option for prevention and treatment of bacterial infection caused by MRS in animals.

This research paper presents the characterization of an enterocin-producing Enterococcus durans MF5 isolate and the determination of the in vitro antilisterial activity of enterocin produced by this isolate, named Ent-MF5. PCR-based screening for bacteriocin biosynthetic genes revealed that E. durans MF5 harbors multiple enterocin-encoding genes (ent A, B, P and X), classified as class II bacteriocins and enterocin-P of Enterococcus faecium (sharing up to 99% similarity at the genetic level). E. durans MF5 is sensitive to eight clinically important antibiotics and does not possess cytolysin activator -cylA, gelatinase -gelE and hyaluronidase -hylA virulence genes. The antilisterial activity of Ent-MF5 was abolished by trypsin, α-chymotrypsin, protease and proteinase-K. Ent-MF5 showed thermal and pH stability. In addition, the activity of Ent-MF5 was unaffected in the presence of various surfactants (1% SDS, Triton X-100, Tween 20, and Tween 80). Ent-MF5 exhibited antimicrobial activity against Listeria monocytogenes, Listeria innocua, Listeria ivanovii and Listeria seeligeri at concentrations as low as 0.13 μg/ml. Ent-MF5 had a bactericidal effect against L. monocytogenes with a significant reduction in surviving cells at concentrations equal to or greater than 0.13 μg/ml. A 75–100% reduction in L. monocytogenes growth and bactericidal effect determined by CFU counts was observed following treatment with Ent-MF5 at 4.47 μg/ml at time points starting at 2 and 4 h, respectively. Ent-MF5 action is associated with Listeria cell membrane damage, as observed by flow cytometry and fluorescence microscopy. Thus, the effective antilisterial activity and stability of Ent-MF5 presents promising perspectives for application as biopreservatives in the food industry.

The aim of this study was to evaluate the efficacy of durancin 61A alone or in combination with nisin, pediocin PA-1, reuterin, microcin J25, vancomycin or tetracycline as an inhibitor of resistant clinical pathogens and to shed light on its mode of action. Durancin and reuterin were effective inhibitors of Clostridium difficile, vancomycin-resistant Enterococcus faecium and methicillin-resistant Staphylococcus aureus. The combination of durancin and reuterin was highly synergistic against C. difficile (fractional inhibitory concentration index = 0.2). Durancin/vancomycin combination was synergistic against S. aureus ATCC 700699 (fractional inhibitory concentration index = 0.3). Conclusion & future perspective: Durancin 61A alone or combined with other bacteriocins or antibiotics may therefore provide a possible therapeutic option for the treatment of infections by these pathogens.

The purpose of this investigation was to compare the performance of species-specific polymerase chain reaction (PCR), matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) and phenotypic identification systems for the identification of Enterococcus species. A total of 132 clinical isolates were investigated by the following: (1) a multiplex real-time PCR assay targeting ddl Enterococcus faecium , ddl Enterococcus faecalis , vanC1 and vanC2 / C3 genes, and a high-resolution melting (HRM) analysis of the groESL gene for the differentiation of Enterococcus casseliflavus and Enterococcus gallinarum ; (2) Bruker MS; (3) VITEK MS; and (4) the VITEK 2 system. 16S rRNA gene sequencing was used as a reference method in the study. The 132 isolates were identified as 32 E. faecalis , 63 E. faecium , 16 E. casseliflavus and 21 E. gallinarum . The multiplex PCR, Bruker MS and VITEK MS were able to identify all the isolates correctly at the species level. The VITEK 2 system could identify 131/132 (99.2 %) and 121/132 (91.7 %) of the isolates at the genus and species levels, respectively. The HRM- groESL assay identified all (21/21) E. gallinarum isolates and 81.3 % (13/16) of the E. casseliflavus isolates. The PCR methods described in the present study are effective in identifying the enterococcal species. MALDI-TOF MS is a rapid, reliable and cost-effective identification technique for enterococci. The VITEK 2 system is less efficient at detecting non- faecalis and non- faecium Enterococcus species.

Lactic acid bacteria (LAB) isolated from different sources (dairy products, fruits, fresh and fermented vegetables, fermented cereals) were screened for antimicrobial activity against other bacteria, including potential pathogens and food spoiling bacteria. Six strains have been shown to produce bacteriocins: Lactococcus lactis 19.3, Lactobacillus plantarum 26.1, Enterococcus durans 41.2, isolated from dairy products and Lactobacillus amylolyticus P40 and P50, and Lactobacillus oris P49, isolated from bors. Among the six bacteriocins, there were both heat stable, low molecular mass polypeptides, with a broad inhibitory spectrum, probably belonging to class II bacteriocins, and heat labile, high molecular mass proteins, with a very narrow inhibitory spectrum, most probably belonging to class III bacteriocins. A synergistic effect of some bacteriocins mixtures was observed. We can conclude that fermented foods are still important sources of new functional LAB. Among the six characterized bacteriocins, there might be some novel compounds with interesting features. Moreover, the bacteriocin-producing strains isolated in our study may find applications as protective cultures.

Probiotic properties of Enterococcus strains have been reported previously. In this study, we investigated the effects of Enterococcus (E.) durans TN-3 on the development of dextran sulfate sodium (DSS) colitis. BALB/c mice were fed with 4.0% DSS in normal chow. Administration of TN-3 (10mg/day) was initiated 7days before the start of DSS feeding. Mucosal cytokine expression was analyzed by real time-PCR and immunohistochemistry. The lymphocyte subpopulation were analyzed by flow cytometry. The gut microbiota profile was analyzed by a terminal-restriction fragment length polymorphism method (T-RFLP). The disease activity index and histological colitis score were significantly lower in the DSS plus TN-3 group than in the DSS group. The mucosal mRNA expression of proinflammatory cytokines (IL-1β, IL-6, IL-17A and IFN-γ) decreased significantly in the DSS plus TN-3 group as compared to the DSS group. The proportion of regulatory T cells (Treg cells) in the mucosa increased significantly in the DSS plus TN-3 group as compared to the DSS group. Both fecal butyrate levels and the diversity of fecal microbial community were significantly higher in the TN-3 plus DSS group than in the DSS group. E. durans TN-3 exerted an inhibitory effect on the development of DSS colitis. This action might be mediated by the induction of Treg cells and the restoration of the diversity of the gut microbiota.