Probiotic Strain Enterococcus Faecium Research Articles

Comparative Efficacy of Parenteral and Mucosal Recombinant Probiotic Vaccines Against SARS-CoV-2 and S. pneumoniae Infections in Animal Models

Background: The accumulation of specific IgG antibodies in blood serum is considered a key criterion for the effectiveness of vaccination. For several vaccine-preventable infections, quantitative indicators of the humoral response have been established, which, when reached, provide a high probability of protection against infection. The presence of such a formal correlate of vaccine effectiveness is crucial, for example, in organizing preventive measures and validating newly developed vaccines. However, can effective protection against infection occur when the level of serum antibodies is lower than that provided by parenteral vaccination? Will protection be sufficient if the same vaccine antigen is administered via mucosal membranes without achieving high levels of specific IgG circulating in the blood? Methods: In this study, we compared the immunogenicity and protective efficacy of parenteral and mucosal forms of vaccines in experimental animals, targeting infections caused by the SARS-CoV-2 coronavirus and Streptococcus pneumoniae. We investigated the protective properties of a fragment of the coronavirus S1 protein administered intramuscularly with an adjuvant and orally as part of the probiotic strain Enterococcus faecium L3 in a Syrian hamster model. A comparative assessment of the immunogenicity and protective efficacy of a recombinant tandem (PSP) of immunogenic peptides from S. pneumoniae surface proteins, administered either parenterally or orally, was performed in a Balb/c mouse model. Results: Both models demonstrated significant differences in the immunogenicity of parenteral and oral vaccine antigens, but comparable protective efficacy.

Comparative analysis of the effectiveness of probiotic enterococcus strains in gastroenteritis in piglets

According to a number of authors, a sharp change in the type of feeding and the effects of stress factors in piglets after weaning lead to disorders not only of metabolism, but also of membrane digestion, which further contributes to a decrease in body resistance and the occurrence of various diseases of the digestive organs.In the course of our study, it was found that the use of probiotic strains Enterococcus faecium L-3 and Enterococcus faecium 1-35 for the treatment of gastroenteritis in piglets during weaning had a similar effect on restoring intestinal enzymatic activity. In piglets that were not treated with probiotics (control 1), enzymatic activity in the digestive tract was only partially normalized during the study. We also evaluated some parameters of the ongoing metabolic processes in the body of weaning piglets by studying the biochemical blood parameters of the experimental groups after prescribing the probiotic strain Enterococcus faecium L-3 (experiment 1) and Enterococcus faecium 1-35 (experiment 2) comparing them with those of the control group of animals.

Fabrication of hyaluronic acid-inulin coated Enterococcus faecium for colon-targeted delivery to fight Fusobacterium nucleatum

The abundance of Fusobacterium nucleatum (F. nucleatum) is highly associated with the development and poor prognosis of colorectal cancer (CRC), which is regarded as a promising target for CRC. However, until now, the novel strategy to clear F. nucleatum in the colon and CRC has not been well proposed. Herein, a probiotic strain Enterococcus faecium (E. faecium, EF47) is verified to secrete various organic acids and bacteriocins to exert superior antimicrobial activity towards F. nucleatum. However, the oral delivery of EF47 is affected by the complex digestive tract environment, so we design the hyaluronic acid-inulin (HA-IN) coated EF47 for colon-targeted delivery to fight F. nucleatum. IN can protect EF47 from the harsh gastrointestinal tract environment and is degraded specifically in the colon, acting as prebiotics to further promote the proliferation of EF47. The exposed HA can also enhance the targeting effect to the tumor area via the interaction with the CD44 receptor on the tumor cells, which is confirmed to increase the adhesive ability in tumor tissues and inhibit the growth of F. nucleatum. Therefore, this colon-targeted delivery system provides a novel platform to realize high-activity and adhesive delivery of probiotics to assist the therapeutic efficiency of CRC.

Antimicrobial activity of probiotic strain Enterococcus faecium 1-35

The study examines the search for alternative solutions to replace feed antibiotics for poultry farming. During the study of the probiotic strain Enterococcus faecium 1-35, its antimicrobial properties against some pathogenic microorganisms were established. Using a bioinformatics approach, genes for non-ribosomal peptide synthetases were discovered in the genome of Enterococcus faecium 1-35. A synthesis product of which is believed to have biological activity. Enterococcus faecium strain 1-35, capable of synthesizing active peptide products, can be regarded as an alternative solution when replacing feed antibiotics. The results obtained require additional studies of practical application to confirm the expected effect.

Development of Bioactive Opuntia ficus-indica Edible Films Containing Probiotics as a Coating for Fresh-Cut Fruit.

Bioactive edible films have received more attention in recent years as a method for food preservation with value-added functions. The aim of this study was to develop a bioactive edible film containing mucilage of cactus (Opuntia ficus-indica) and incorporating the probiotic strain Enterococcus faecium FM11-2 as an active component to promote consumer health benefits. Opuntia ficus-indica is rich in nutritional and bioactive compounds and the abundance of this cactus makes it attractive for food applications. Mucilage of Opuntia ficus-indica contained 0.47 ± 0.06 mg/g total sugar, 0.33 ± 0.06 mg AGE/mL phenolic content, 0.14 mg/ mL vitamin C, and possessed 35.51 ± 1.88% DPPH scavenging activity. The edible film that was developed exhibited the following characteristics: thickness of 0.02-0.11 mm, percent moisture content 0.19-0.24%, water solubility 30.66-59.41% and water vapor permeability of 0.15-1.5 g·mm/m2·min·kpa, while the range of the variation depended on the type of plasticizer used (either sorbitol or glycerol). The addition of sorbitol in the film provided the maximum mechanical strength based on the evaluation of tensile strength, Young's modulus and elongation at break (44.71 ± 0.78 MPa, 113.22 ± 0.23 MPa and 39.47 ± 0.61%, respectively). The optimal formulation of the edible film, according to the physicochemical, physical and maintenance of fresh-cut apple slices, contained cactus mucilage, gelatin, glycerol and a probiotic. The incorporation of a probiotic into the cactus film created a bioactive edible film that could provide a health benefit. While improvement is needed to maintain the survival rate of the probiotic, this work presents an exciting method for furthering the study of food preservation with edible films.

A virulence factor as a therapeutic: the probiotic Enterococcus faecium SF68 arginine deiminase inhibits innate immune signaling pathways

ABSTRACT The probiotic bacterial strain Enterococcus faecium SF68 has been shown to alleviate symptoms of intestinal inflammation in human clinical trials and animal feed supplementation studies. To identify factors involved in immunomodulatory effects on host cells, E. faecium SF68 and other commensal and clinical Enterococcus isolates were screened using intestinal epithelial cell lines harboring reporter fusions for NF-κB and JNK(AP-1) activation to determine the responses of host cell innate immune signaling pathways when challenged with bacterial protein and cell components. Cell-free, whole-cell lysates of E. faecium SF68 showed a reversible, inhibitory effect on both NF-κB and JNK(AP-1) signaling pathway activation in intestinal epithelial cells and abrogated the response to bacterial and other Toll-like receptor (TLR) ligands. The inhibitory effect was species-specific, and was not observed for E. avium, E. gallinarum, or E. casseliflavus. Screening of protein fractions of E. faecium SF68 lysates yielded an active fraction containing a prominent protein identified as arginine deiminase (ADI). The E. faecium SF68 arcA gene encoding arginine deiminase was cloned and introduced into E. avium where it conferred the same NF-κB inhibitory effects on intestinal epithelial cells as seen for E. faecium SF68. Our results indicate that the arginine deiminase of E. faecium SF68 is responsible for inhibition of host cell NF-κB and JNK(AP-1) pathway activation, and is likely to be responsible for the anti-inflammatory and immunomodulatory effects observed in prior clinical human and animal trials. The implications for the use of this probiotic strain for preventive and therapeutic purposes are discussed.

The use of <i>E. faecium</i> probiotic and autoprobiotic in patients with type 2 diabetes mellitus

BACKGROUND: Bacteria in the gastrointestinal tract regulate eating behavior and metabolic processes. A number of probiotic strains have a positive effect on glucose and insulin metabolism. A series of studies have demonstrated the effectiveness of autoprobiotic therapy in the treatment of patients with gastrointestinal diseases. The efficacy and safety of an indigenous strain of Enterococcus faecium in overweight patients with type 2 diabetes has not been studied in the past.
 AIM: To evaluate the effectiveness of products containing the probiotic strain Enterococcus faecium L3 or the indigenous strain Enterococcus faecium in patients with diabetes mellitus (DM) type 2.
 MATERIALS AND METHODS: Patients with diabetes mellitus type 2 were randomized into two groups: group of probiotic therapy with an industrial strain of Enterococcus faecium L3 (11 patients); group of autoprobiotic therapy based on an indigenous strain of Enterococcus faecium (9 patients). Therapy was for 14 days. Before and 10-14 days after the end of therapy, anthropometric parameters were assessed; psychometric testing was carried out; biochemical parameters of blood serum were studied. The intestinal microbiota was studied by real-time polymerase chain reaction.
 RESULTS: Probiotic therapy based on Enterococcus faecium L3 and autoprobiotic therapy based on indigenous Enterococcus faecium had no significant effect on glucose metabolism in patients with type 2 DM. In the Enterococcus faecium L3 group and the group of indigenous Enterococcus faecium there was a significant decrease in gastroenterological complaints on scales of the GSRS questionnaire. Patients in both groups showed a statistically significant decrease in the total bacterial mass, an increase in the quantitative content of Lactobacillus spp., a decrease in the population of Bifidobacterium spp. and Bacteroides thetaiotaomicron.
 CONCLUSIONS: The probiotic strain Enterococcus faecium L3 and the indigenous Enterococcus faecium do not have a significant effect on the metabolism of glucose and insulin, while they contribute to a similar change in the component composition of the microbiota and a decrease in the severity of gastroenterological complaints.

Developing a Live Probiotic Vaccine Based on the Enterococcus faecium L3 Strain Expressing Influenza Neuraminidase.

Probiotic microorganisms are currently considered as a promising platform for the development of recombinant vaccines expressing foreign antigens. In this study, we generated and evaluated the live mucosal recombinant vaccine by integrating genes encoding influenza virus neuraminidase (NA) of the N2 subtype into the DNA of the probiotic strain Enterococcus faecium L3 (L3). We confirmed NA expression in the pili of L3 using immune electron microscopy. Mice were fed with a probiotic vaccine containing the NA gene (L3-NA) or pure L3. Oral administration of L3-NA caused detectable increase in virus-specific serum IgG and local IgA after the third feeding. Immunization with L3-NA increased the survival rate by 34% when the mice were infected using A(H1N1)pdm09 influenza virus after the third feeding. After S. pneumoniae post-influenza infection, the L3-NA-immunized mice were 50% more protected from lethality in comparison with L3-fed mice. Thus, a live probiotic vaccine candidate based on L3 induced the formation of systemic and local immunity and provide partial protection against complicated influenza.

Development of a potentially functional chocolate spread containing probiotics and structured triglycerides

New functional hazelnut and chocolate spreads, containing the probiotic strain Enterococcus faecium CRL 183 and different concentrations of medium-long-medium (MLM)-type structured triacylglycerols (TAGs) were developed. TAGs did not alter the viability of the probiotic strain, during 180 days of storage at 20 °C. The formulations presented similar contents of proteins, lipids, and carbohydrates, water activity below 0.5, thermal stability and were microbiologically stable. The decrease of saturated fat had a negative effect on the rheological properties of the spreads, with reduction on viscosity, Casson yield value and texture parameters. However, only the spread with 75% substitution of vegetable fat by structured TAG showed severe reduction in rheological properties that affected the product spreadability. The use of probiotic strain and replacement of fat (50%) by MLM-type TAG resulted in a new functional chocolate spread with significant increment in medium-chain and unsaturated fatty acids.

Determination of the biosafety of potential probiotic Enterococcus faecalis and Enterococcus faecium strains isolated from traditional white cheeses

The aims of this study were to characterize antibiotic resistance, determination of genotypic and phenotypic virulence characters; and to evaluate the safety of 12 Enterococcus faecalis and 8Enterococcus faecium strains isolated from Iranian and Turkish white cheeses. All strains were researched for production capability of cytolysin and gelatinase, biofilm formation and sensitivity against ten antibiotics. The presence of vancomycin resistance genes and virulence factors genes like cylA and cylB, esp, agg, asa1, EfaAfs, EfaAfm, cob, ccf, and hly were determined by polymerase chain reaction and sequence analysis. All the investigated strains were resistant to at least one class of antibiotic, and 40% of the strains were classified as multi drug resistant. E. faecalis RI 11, RI 17, RI 21 strains demonstrated both gelatinase and biofilm activities. Moreover, there was a significant correlation between gelatinase activity and the presence of gelE (P < 0.05). While ccf and asa1 genes were determined in all strains, efaAfm and clyB genes were not found in any strains. E. faecium RI 56, RI 58 and RI 59 strains were found to contain different molecular sizes of plasmid DNA. The phenotypic and genotypic characteristics of E. faecalis and E. faecium strains were determined in terms of functional and safety perspectives.

Effective microencapsulation of Enterococcus faecium in biopolymeric matrices using spray drying.

The objective of this work was to evaluate the potential of whey protein concentrate (WPC), native agave fructans (NAF), and their mixture (WPC-NAF, 1:1 w/w) as wall materials and evaluate the physicochemical properties and stability of encapsulated Enterococcus faecium during the spray drying, storage, and passage through the simulated gastrointestinal tests. The encapsulated microorganisms with WPC-NAF by spray drying showed greater viability (9.26 log CFU/g) and a higher microencapsulation yield (88.43%). They also had a smaller reduction in the cell count (0.61 log cycles), while the microcapsules produced with NAF had the greatest reduction in viability during the simulated gastrointestinal tests. Similarly, probiotics encapsulated with WPC-NAF revealed a higher survival rate (> 8 log CFU/g) when stored at a water activity of 0.328. The thermal analysis showed that the addition of NAF to the WPC produced a slight shift in the Tg towards temperatures higher than that shown by NAF. Therefore, this study provides evidence that the spray drying process was appropriate to encapsulate the probiotic strain Enterococcus faecium and that the mixture WPC-NAF protected it from adverse drying conditions and improved the viability of Enterococcus faecium during storage and simulated gastrointestinal tests, demonstrating that the combination of NAF and WPC as encapsulating material is adequate in the production of more stable microcapsules with potential application in various foods.Key Points• E. faecium was successfully encapsulated in WPC and NAF.• WPC-NAF offered protection to E. faecium in the gastrointestinal tests and during storage.• Aw around 0.328 positively influenced the viability of the microorganism during storage. Graphical abstract.

Draft Genome Sequences of Five Potentially Probiotic Enterococcus faecium Strains Isolated from an Artisanal Tunisian Meat (Dried Ossban).

Here, we report the first draft genome sequences of five bacteriocinogenic and potentially probiotic Enterococcus faecium strains (MZF1 to MZF5), which were isolated from homemade Tunisian meat (dried ossban). The estimated median genome sizes were about 2,582,641 ± 109,039 bp, with a median G+C content of 40% ± 0.4%.

Development of experimental pneumococcal vaccine for mucosal immunization.

Streptococcus pneumonia is an important human pathogen that causes various severe diseases such as pneumonia, otitis and meningitis. Vaccination against S. pneumoniae is implemented in many developed countries. The presently used vaccines are safe, well tolerated but relatively expensive and require modification due to the immunological changes of the epidemic strains. This paper describes the development of a new pneumococcal vaccine candidate for immunization on mucosal surfaces. For this purpose the antigens of chimeric protein PSPF, previously suggested for an injectable S. pneumoniae vaccine, were expressed on the surface of the live probiotic strain Enterococcus faecium L3. Experiments on laboratory mice vaccinated with live bacteria demonstrated the appearance of the specific IgA and IgG which provide protection against the lethal S. pneumoniae infection.

Post hatch recovery of a probiotic Enterococcus faecium strain in the yolk sac and intestinal tract of broiler chickens after in ovo injection

ABSTRACTConcerns about antibiotic-resistant bacteria and their presence in animal products grow and thus alternatives to use of antibiotics in animal production are being investigated. Probiotics have gained increased focus due to improvements in performance, immune health and pathogen reduction when provided to poultry through feed. These traits may be further improved if probiotics can be provided to the embryo before hatch, before meeting environmental pathogens. The objective was to determine the faith of a probiotic Enterococcus faecium (M74) strain in the yolk sac and intestinal tract of broiler chickens after injection into hatching eggs. E. faecium M74 (1.4 × 107 CFU/egg) was applied in ovo at day 18 of incubation. From 1- and 7-day-old chickens, 20 samples from yolk sac, caecal tonsils and rest of the intestinal tract were subjected to CFU counting. Isolates from a sample subset were typed by pulsed-field gel electrophoresis (PFGE). Enterococci were found in varying numbers: 1.0 × 104–2.2 × 1010 CFU/g. The prevalence of M74 PFGE profiles was high in 1-day-old (88%) and 7-day-old chickens (67%). This demonstrates that the embryos ingested M74 before hatching, that M74 is viable for intestinal colonization through in ovo administration, and that the strain multiplies in the chickens gastrointestinal tract post hatching.

Post hatch recovery of a probiotic Enterococcus faecium strain in the yolk sac and intestinal tract of broiler chickens after in ovo injection.

Concerns about antibiotic-resistant bacteria and their presence in animal products grow and thus alternatives to use of antibiotics in animal production are being investigated. Probiotics have gained increased focus due to improvements in performance, immune health and pathogen reduction when provided to poultry through feed. These traits may be further improved if probiotics can be provided to the embryo before hatch, before meeting environmental pathogens. The objective was to determine the faith of a probiotic Enterococcus faecium (M74) strain in the yolk sac and intestinal tract of broiler chickens after injection into hatching eggs. E. faecium M74 (1.4×107 CFU/egg) was applied in ovo at day 18 of incubation. From 1- and 7-day-old chickens, 20 samples from yolk sac, caecal tonsils and rest of the intestinal tract were subjected to CFU counting. Isolates from a sample subset were typed by pulsed-field gel electrophoresis (PFGE). Enterococci were found in varying numbers: 1.0×104-2.2×1010 CFU/g. The prevalence of M74 PFGE profiles was high in 1-day-old (88%) and 7-day-old chickens (67%). This demonstrates that the embryos ingested M74 before hatching, that M74 is viable for intestinal colonization through in ovo administration, and that the strain multiplies in the chickens gastrointestinal tract post hatching.

Potential use of probiotic Enterococcus faecium and Enterococcus durans strains in Izmir Tulum cheese as adjunct culture.

In this research, Enterococcus species with probiotic properties were isolated from raw milk and some traditional dairy products and these species were identified by using biochemical, phenotypical and genotypic methods and their potential use in Izmir Tulum cheese production as adjunct culture were investigated. Among the isolated and identified Enterococcus species, three Enterococcus faecium and one Enterococcus durans strains with no vancomycin resistance, no decarboxylation activity, high antimicrobial activity and resistance to acidity were selected, and a mix culture was produced. This culture was mixed with commercial cheese culture containing Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. cremoris in different proportions and used in Izmir Tulum cheese production. Cheese samples were left to ripening for 180days, and their physical, chemical, microbiological and sensory properties were evaluated with comparison to prior studies. Evaluations showed that using adjunct culture in Izmir Tulum cheese production has no negative effect on physical, chemical and microbiological properties and higher proportions of probiotic Enterococci in the culture have a positive effect on the sensory properties, especially on flavor.

Heavy Metals in some Meat Products and Its Biocontrol by Probiotic Strain Enterococcus Faecium (M74) in Minced Meat

Heavy Metals in some Meat Products and Its Biocontrol by Probiotic Strain Enterococcus Faecium (M74) in Minced Meat

Effects of a pathogenic ETEC strain and a probiotic Enterococcus faecium strain on the inflammasome response in porcine dendritic cells

Dendritic cells (DC) are crucial for maintaining intestinal homeostasis and generating proper immune responses to bacteria occurring in the gut. Microbial stimuli can be recognized by intracellular receptors called inflammasomes, e.g., nucleotide oligomerization domain (NOD)-like receptor protein 3 (NLRP3). The aim of the present study was to unravel the inflammasome response of porcine monocyte-derived DC (MoDC). We investigated the capacity of probiotic Enterococcus faecium NCIMB 10415 (E. faecium) and enterotoxigenic Escherichia coli (ETEC) to elicit inflammasome activation. Since inflammasome activation normally requires a two-step process, MoDC were initially incubated with lipopolysaccharide (LPS) in order to prime cells. Primed and unprimed cells were then stimulated with the aforementioned bacterial strains. We also assessed whether preincubation with the probiotic prior to ETEC infection modified the immune response via the inflammasome pathway.Phenotypical analysis by flow cytometry showed that monocytes and MoDC expressed the surface markers CD14, CD16, and CD1 continuously, whereas swine leucocyte antigen (SLA) II was upregulated during differentiation.Following LPS priming, NLRP3, interleukin (IL)-1β and IL-18 mRNA expression, and IL-1β protein release increased. In unprimed cells, ETEC upregulated the expression of inflammasome components at later time points than in LPS-primed MoDC. Preincubation with the probiotic did not influence NLRP3 inflammasome activation in comparison with cells infected with ETEC alone.We conclude that ETEC, but not E. faecium, was able to stimulate inflammasome components in porcine MoDC. The present experimental conditions revealed no NLRP3 inflammasome-dependent protective effects of E. faecium during a pathogenic ETEC challenge.

Enterococcus faecium L-3 in Eradication of Helicobacter pylori: In-vivo and In-vitro

Usage of probiotics as component of anti-helicobacter therapy had been recently approved as a promising trend in the clinical practice. Over the years, we observed high efficacy of probiotic multi bacteriocinogenic strain Enterococcus faecium L-3 as an additional component of the standard triple therapy in eradication of Helicobacter pylori (H.p.) in patients with chronic gastritis associated with H. pylori infection. The cultivation of H.p. in the presence of live probiotic cultures of E. faecium L-3, Escherichia coli M17, Lactobacillus plantarum 8R-A3 showed superior antagonistic activity of enterococci relatively to other tested bacteria. The anti-helicobacter activity of chemically synthesized enterococcal bacteriocins EntB and Enxβ were studied using 6 strains of H. pylori. The strain-specific action of synthetic enterocins used was determined. Testing of sensitivity of H. pylori. to probiotics and their metabolites in vitro would be important for the selection and prediction of the clinical effectiveness of probiotics as well as development of new anti-helicobacter drugs and lines of combined therapies based on enterocins.

Pilot experiment in chickens challenged with Campylobacter jejuni CCM6191 administered enterocin M-producing probiotic strain Enterococcus faecium CCM8558 to check its protective effect

Campylobacter jejuni is one of the most common food-borne pathogens and chickens are the main source of these bacteria. Enterococcus faecium AL41, enterocin M-producing strain (deponed to the Czech Culture Collection of Microorganisms, Brno, Czech Republic – CCM8558) is our isolate previously applied e.g. in broiler rabbits with beneficial effect. In this study it was used in a 11-day experiment with chickens (1-day-old, breed Cobb 500, n = 40) challenged with C. jejuni. Birds had free access to feed and water; they were randomly divided into four groups per 10 chicks each: control (CG), E. faecium CCM8558 (EG1), CCM8558 + C. jejuni CCM6191 (EG2), CCM6191 (EG3). E. faecium CCM8558 (10&lt;sup&gt;9 &lt;/sup&gt;CFU/ml, 200 µl) in Ringer solution was administered daily per os to EG1 and EG2 for 7 days (from day 0–1). EG3 and EG2 were infected individually per os (day 4, CCM6191, 10&lt;sup&gt;8&lt;/sup&gt; CFU/ml in Ringer solution, 200 µl). For microbiota evaluation, faecal mixtures (n = 5) were sampled on day 0–1 (1&lt;sup&gt;st&lt;/sup&gt; sampling), on day 7 (2&lt;sup&gt;nd&lt;/sup&gt; sampling), and on day 11 (3&lt;sup&gt;rd&lt;/sup&gt; sampling) (day 7 of CCM8558 application, day 4 post-infection; day 4 of CCM8558 cessation, day 7 post-infection). Five birds from each group were sacrificed. CCM8558 sufficiently colonized chickens. In faeces of EG2 a tendency to reduce Campylobacter spp. (day 7 of application, day 4 of infection) was noted compared to EG2 (day 11, day 4 of cessation, day 7 post-infection; difference 1.21 log cycles), while in EG3 CCM6191 strain was not reduced. Phagocytic activity (PA) values were significantly higher in infected groups compared to CG and EG1. A significant increase in PA was also noted in EG2 and EG3 at the end of experiment compared to CG or EG1. The strain additive did not evoke oxidative stress. Biochemical parameters were influenced to the reference levels.