Probiotic Enterococcus Faecium Research Articles

Pharmacological Effects of Marine-Derived Enterococcus faecium EA9 against Acute Lung Injury and Inflammation in Cecal Ligated and Punctured Septic Rats.

Microorganisms obtained from the marine environment may represent a potential therapeutic value for multiple diseases. This study explored the possible protective role of marine-derived potential probiotic Enterococcus faecium EA9 (E. faecium) against pulmonary inflammation and oxidative stress using the cecal ligation and puncture (CLP) model of sepsis in Wistar rats. Animals were pretreated with E. faecium for 10 days before either sham or CLP surgeries. Animals were sacrificed 72 hours following the surgical intervention. The histological architecture of lung tissues was evaluated as indicated by the lung injury score. In addition, the extend of pulmonary edema was determined as wet/dry weight ratio. The inflammatory cytokines were estimated in lung tissues, including tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and interleukin-1 beta (IL-1β) using the enzyme-linked-immunosorbent-assay (ELISA) technique. Moreover, markers for lipid peroxidation such as thiobarbituric acid reaction substances (TBARs), and endogenous antioxidants, including reduced glutathione (GSH) were determined in lung tissues. Finally, the enzymatic activities of antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione reductase (GR) were assayed in the lungs. Pretreatment with E. faecium markedly attenuated CLP-induced lung injury and pulmonary edema. Markers for inflammation, including TNF-α, IL-6, and IL-1β were augmented in the lung tissues of CLP animals, while E. faecium ameliorated their augmented levels. E. faecium pretreatment also restored the elevated TBARS levels and the prohibited CAT, SOD, and GPx enzymatic activities in CLP animals. GSH levels were corrected by E. faecium in CLP animals. The inflammatory and lipid peroxidation mediators were positively correlated, while antioxidant enzymatic activities were negatively correlated with CLP-induced lung injury and pulmonary edema. Collectively, marine-derived Enterococcus faecium EA9 might be considered as a prospective therapeutic tool for the management of pulmonary dysfunction associated with sepsis.

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.

Prebiotic and synbiotic effect on rumen papilla length development and rumen pH in 12-week-old calves.

Background and Aim:Europe and the USA have banned antibiotics use as growth promoters. There is a need for alternative products that can ensure production and health protection. Prebiotics has been proposed as alternatives because these materials have wide-ranging physiological effects on gut function, activity of the large intestinal microflora, mineral absorption, and immunity. The aim of this study was to determine the effect of three different doses of inulin, a prebiotic, in combination with probiotic Enterococcus faecium (a new synbiotic) on postnatal rumen development by comparing rumen papilla length, width, muscle layer thickness, and content pH level.Materials and Methods:Randomly selected 23 (±5)-days-old healthy male Holstein crossbreed calves, weighing 50 kg (±5 kg), were randomly allocated to seven groups, ten in each group. The calves were kept in a pen of 5, under the same conditions and were fed twice a day, ~3.5 liters of whole milk per feeding. Control group (C n=10) was fed with whole milk only (no additives were added). The six other groups (three prebiotics and three synbiotics) received food additives with their morning milk feeding. The source of prebiotics, Jerusalem artichoke powder concentrate (JAPC) contained 50% of inulin. JAPC in doses of 6 g, 12 g, or 24 g were added to the milk. Formed prebiotic groups were denoted as PreG6, PreG12, and PreG24. To evaluate if the addition of the probiotic E. faecium 2×109 colony forming unit g−1 to manufacturer recommended dose of 0.25 g improves inulin effect on rumen, it was added to all their JAPC doses. The new content synbiotic groups were denoted as SynG6, SynG12, and SynG24. On day 57 of the study, when all calves were approximately 12 weeks old, they were slaughtered in a certified slaughterhouse. Tissue cultures for histological analysis were obtained from Saccus dorsalis and Saccus ventralis of the rumen. Tissue culture staining for histology was carried out using hematoxylin and eosin staining method. Rumen histological samples were used to measure papilla length, width, and muscle layer thickness. Each sample was used to make five measurements on the present rumen papilla.Results:The results showed that by adding 12 g of inulin to whole milk when feeding calves improves rumen papilla development, which is seen by increased length and width of papilla, especially in the Saccus ventralis region. By combing this dose of inulin with 0.25 g of E. faecium, a significant increase of papilla is achieved. Saccus ventralis muscle layer in the rumen is thicker than it is in Saccus dorsalis regardless of addition of prebiotics or synbiotics.Conclusion:The addition of inulin to whole milk can influence the pH of the rumen by making it more alkaline. The addition of prebiotic inulin and a novel synbiotic (inulin combined with E. faecium) can accelerate postnatal rumen development and improve its functionality.

Potential Probiotic Enterococcus faecium OV3-6 and Its Bioactive Peptide as Alternative Bio-Preservation.

Probiotic Enterococcus faecium OV3-6 and its secreted active peptide were characterized and investigated. The strain survived in simulated gastric and small intestinal conditions at 88.16% and 94.33%, respectively. The safety assessment revealed that the strain was shown α-hemolysis and susceptible to most clinically relevant antibiotics, but intermediate sensitivity to erythromycin and kanamycin was found. It does not harbor any virulence genes except for the efaAfm gene. Both of its living cells and the cell-free supernatants (CFS) of the strain significantly reduced the adhesion of E. coli and S. Typhi on Caco-2 cells. The strain can regulate the secretion of pro and inflammatory cytokines, IL-6 and IL-12 and induce the secretion of anti-inflammatory IL-10 of the Caco-2 cell. The strain can prevent the growth of Gram-positive strains belonging to the genera Bacillus, Carnobacterium, Listeria, and Staphylococcus. It also presented the entP gene that involves the production of bacteriocin named enterocin P. The antimicrobial peptide was matched 40% with 50S ribosomal proteins L29 (7.325 kDa), as revealed by LC-MS/MS. This active peptide exhibits heat stability, is stable over a wide pH range of 2−10, and maintains its activity at −20 and 4 °C for 12 weeks of storage. Altogether, E. faecium OV3-6 thus has potential for consideration as a probiotic and bio-preservative for applied use as a fermented food starter culture and in functional food or feed industries.

Enterococcus faecium HDRsEf1 inhibits Lipopolysaccharide-induced downregulation of zona occludens -1 expression via toll-like receptor 2/4-mediated c-Jun N-terminal kinase/activator protein-1 signalling pathways.

Zona occludens-1 (ZO-1) is a key regulatory tight junction protein that plays an important role in maintaining gastrointestinal health. In this study, we investigated the protective effect and regulation mechanism of the probiotic Enterococcus faecium HDRsEf1 on tight junction protein ZO-1 at the cellular and molecular levels. We established lipopolysaccharide (LPS)-induced intestinal epithelial cell injury model and detected the protective effect of HDRsEf1 on ZO-1 in IPEC-J2 cells by real-time polymerase chain reaction and Western blot. The results showed that HDRsEf1 inhibited the downregulation of ZO-1 expression induced by LPS. HDRsEf1 stabilized the destruction of the ZO-1 structure caused by LPS in an immunofluorescence assay. Through gene overexpression and siRNA interference tests, we found that transcription factor activator protein 1 (AP-1) inhibited the level of ZO-1 expression. Silencing experiment further supported that the protective effect of HDRSEF1 might be mediated by suppression of LPS-provoked activation of apoptosis signal-regulating kinase 1 (ASK1)/mitogen-activated protein kinase kinase 7 (MKK7)/c-Jun N-terminal kinase (JNK) signalling pathways. In addition, HDRsEf1 could stabilize ZO-1 expression by increasing toll-like receptor 2 (TLR2) expression and competing with LPS for the TLR4 binding site. More interestingly, we also found that HDRsEf1 could stabilize ZO-1 expression through inhibiting the production of tumour necrosis factor-α (TNF-α) induced by LPS. HDRsEf1 could protect the IPEC-J2 cell against LPS induced downregulation of ZO-1 expression by inhibiting the activation of TLR2/4-mediated JNK-AP-1 and signalling cascade and the production of TNF-α. This study can provide a theoretical basis for probiotics to regulate the expression of intestinal tight junction proteins, and supply technical support for probiotics to prevent and treat animal intestinal infectious diseases.

Serotonin Exposure Improves Stress Resistance, Aggregation, and Biofilm Formation in the Probiotic Enterococcus faecium NCIMB10415

The role of the microbiota–gut–brain axis in maintaining a healthy status is well recognized. In this bidirectional flux, the influence of host hormones on gut bacteria is crucial. However, data on commensal/probiotics are scarce since most reports analyzed the effects of human bioactive compounds on opportunistic strains, highlighting the risk of increased pathogenicity under stimulation. The present investigation examined the modifications induced by 5HT, a tryptophan-derived molecule abundant in the intestine, on the probiotic Enterococcus faecium NCIMB10415. Specific phenotypic modifications concerning the probiotic potential and possible effects of treated bacteria on dendritic cells were explored together with the comparative soluble proteome evaluation. Increased resistance to bile salts and ampicillin in 5HT-stimulated conditions relate with overexpression of specific proteins (among which Zn-beta-lactamases, a Zn-transport protein and a protein involved in fatty acid incorporation into the membrane). Better auto-aggregating properties and biofilm-forming aptitude are consistent with enhanced QS peptide transport. Concerning interaction with the host, E. faecium NCIMB10415 enhanced dendritic cell maturation, but no significant differences were observed between 5HT-treated and untreated bacteria; meanwhile, after 5HT exposure, some moonlight proteins possibly involved in tissue adhesion were found in higher abundance. Finally, the finding in stimulated conditions of a higher abundance of VicR, a protein involved in two-component signal transduction system (VicK/R), suggests the existence of a possible surface receptor (VicK) for 5HT sensing in the strain studied. These overall data indicate that E. faecium NCIMB10415 modifies its physiology in response to 5HT by improving bacterial interactions and resistance to stressors.

Orodispersible Film Loaded with Enterococcus faecium CRL183 Presents Anti-Candida albicans Biofilm Activity In Vitro.

Background: Probiotic bacteria have been emerging as a trustworthy choice for the prevention and treatment of Candida spp. infections. This study aimed to develop and characterize an orodispersible film (ODF) for delivering the potentially probiotic Enterococcus faecium CRL 183 into the oral cavity, evaluating its in vitro antifungal activity against Candida albicans. Methods and Results: The ODF was composed by carboxymethylcellulose, gelatin, and potato starch, and its physical, chemical, and mechanical properties were studied. The probiotic resistance and viability during processing and storage were evaluated as well as its in vitro antifungal activity against C. albicans. The ODFs were thin, resistant, and flexible, with neutral pH and microbiologically safe. The probiotic resisted the ODF obtaining process, demonstrating high viability (>9 log10 CFU·g−1), up to 90 days of storage at room temperature. The Probiotic Film promoted 68.9% of reduction in fungal early biofilm and 91.2% in its mature biofilm compared to the group stimulated with the control film. Those results were confirmed through SEM images. Conclusion: The probiotic ODF developed is a promising strategy to prevent oral candidiasis, since it permits the local probiotic delivery, which in turn was able to reduce C. albicans biofilm formation.

Optimization of cryoprotectants for freeze-dried potential probiotic Enterococcus faecalis and evaluation of its storage stability

Probiotic application remains a challenge since decades, as microorganisms lose their viability and functionality during the manufacture and storage of freeze-dried powders. Appropriate cryoprotectants are important in achieving particular probiotic preparation. Recently, Enterococcus spp. such as Enterococcus faecalis and E. faecium have been used as probiotics for human or animal health promotion. We aimed to determine a suitable protectant to protect the viability and pathogen inhibition potential of E. faecalis probiotic strains during freeze-drying and subsequent storage. E. faecalis probiotic strains were lyophilized in the presence of glucose, fructose, sucrose, skim milk, and distilled water. The lyophilized vials were stored at 4 °C for up to 30 days. Then, the cells from these vials were evaluated for their probiotic functionality. Among several protectants, skim milk exhibited highly protective capability to protect E. faecalis cell viability during freeze-drying and 30-day storage, with survival rates of E. faecalis strains ranging from 95.96 ± 0.20% to 103.42 ± 1.96% and 95.68 ± 0.72% to 102.95 ± 2.12%, respectively. The presence of skim milk could also effectively enhance the antimicrobial activity of all freeze-dried E. faecalis strains against toxigenic Clostridium difficile strains and maintain their activity after 30-day storage. Furthermore, hemolytic analysis demonstrated the safety of these freeze-dried E. faecalis strains. Overall, the results indicate that skim milk could be used as a suitable cryoprotective substance for freeze-dried E. faecalis probiotics, with potential applications in functional food, medicine, and feed industries.

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.

Heat Resistance of Candidate Probiotic Enterococcus faecalis R22B in Different Matrices

Fermented fish, including rusip and bekasam, contains lactic acid bacteria, some of which have probiotic properties. Considering the beneficial effect, the count of viable cells in probiotic carrier foods must be at least 106 CFU/g throughout the product shelf-life. The processing and storage condition significantly affect the viability of probiotics in carrier foods. This research was conducted to determine the heat resistance of probiotic candidate Enterococcus faecalis R22B isolated from rusip in three different media: bekasam, rusip, and physiological saline solution (NaCl 0.85%). E. faecalis R22B was heated in each medium at a temperature of 70 °C for 0, 2, 4, 8, and 12 min. The media characteristics showed differences within the parameters included pH values of 4.41-6.99; aw 0.67-0.92; moisture content 72.56-98.09%; ash content 0.69-12.32%; protein content 0-14.7%; fat content 0-0.84%; and carbohydrate content from 0-4.93%. The results showed that the difference in the heating medium had no significant effect (p&gt;0.05) on the heat resistance of the bacteria, as indicated by the D70 value. The required average heating time at 70 oC to decrease E. faecalis R22B cells by 1log10 (D70 value) in bekasam, rusip, and physiological saline media were 3.17, 3.29, and 3.55 min, respectively. Despite having a lower DT-value, the reduction of bacteria cells in three media heated at 70 oC showed no difference (p&gt;0.05); therefore, bekasam and rusip are suitable for probiotic carrier food.

IN VITRO ANTIOXIDANT AND IN VIVO ANTIDIABETIC ACTIVITY OF TWO POTENTIAL PROBIOTIC ENTEROCOCCUS SPP. ON ALLOXAN-INDUCED DIABETIC RATS

Objective: In vitro antioxidant activity, in vivo antidiabetic property and intestinal attachment by two potential probiotic bacterial strains, namely, Enterococcus faecium and Enterococcus hirae were studied using albino rats.&#x0D; Methods: Antioxidant the activity was assessed using 2,2-Diphenyl-1-picrylhydrazyl radicals scavenging assay. Alloxan was administered intraperitoneally to induce diabetic conditions in experimental rats. Animals were treated with oral administration of Enterococcus spp., such as E. faecium, and E. hirae isolated from goat and sheep milk. The control animal group received normal saline for the same days. Glibenclamide drug was used as a positive control against probiotic bacterial cells.&#x0D; Results: However, administration of probiotic bacterial strains E. faecium and E. hirae, in albino rats significantly (p&lt;0.05) at varying doses lowered blood glucose levels in diabetic rats as compared to the diabetic control group. Both the species of Enterococcus increased the bodyweight of experimental rats. However, E. faecium was the best antidiabetic strain having the antioxidant activities also in comparison to E. hirae. The attachment of probiotic bacterial cells E. faecium on the rat’s intestine wall against pathogens was examined. Furthermore, E. faecium showed its aggregation with pathogens by attachment of the intestines of albino rats. This showed that both the bacterial strains exhibited in vivo antidiabetic effect.&#x0D; Conclusion: The results of this study showed that probiotic bacteria possess antioxidant, antidiabetic activities, and attachment of intestine.

Effect of a Synbiotic Supplement on Fear Response and Memory Assessment of Broiler Chickens Subjected to Heat Stress.

Simple SummaryHeat stress is a serious environmental problem, challenging poultry health and welfare globally, especially during summer season. The breeding program of faster-growing broiler chickens affects their biological homeostasis, causing structural and functional damage to the brain, leading to mental disorders. Improvement in the gut microbiota with synbiotic dietary supplements has become a useful biotherapeutic method for treating various diseases, including neuroinflammation-induced mental illness and memory damage. Therefore, this study aimed to assess the effect of a dietary synbiotic supplement on fear response and memory assessment in heat-stressed broiler chickens. We used 360 1-day-old broiler chicks and assigned them to one of three dietary treatments: a regular diet mixed with a synbiotic containing a probiotic (Enterococcus faecium, Pediococcus acidilactici, Bifidobacterium animalis, and Lactobacillus reuteri) and a prebiotic (fructooligosaccharides) at 0, 0.5, and 1.0 g/kg. Object memory, touch, novel object, isolation, and tonic immobility tests were conducted at relative days of age. At 42 day, blood was collected for detecting corticosterone and tryptophan concentrations and examining heterophile/lymphocyte ratios. The data suggest that the synbiotic-reduced heat-stress responses and related emotional disorder may be mainly caused by increasing the activation of the serotonergic system via the microbiota–gut–brain axis. The aim of this study was to evaluate the effect of a synbiotic containing a probiotic (Enterococcus faecium, Pediococcus acidilactici, Bifidobacterium animalis, and Lactobacillus reuteri) and a prebiotic (fructooligosaccharides) on fear response, memory assessment, and selected stress indicators in broilers subjected to heat stress. A total of 360 1-day-old Ross 708 chicks were evenly divided among three treatments: a basal diet mixed with a synbiotic at 0 (G-C), 0.5 (G-0.5X), and 1.0 (G-1.0X) g/kg. After 15 d, the broilers were exposed to 32 °C for 9 h daily until 42 d. The object memory test was conducted at 15 day; touch, novel object, and isolation tests were conducted at 35 day; tonic immobility (TI) took place at 41 day. At 42 day, plasma corticosterone and tryptophan concentrations and heterophile/lymphocyte (H/L) ratios were measured. Compared to controls, synbiotic-fed broilers, regardless of concentration, had a shorter latency to make the first vocalization, with higher vocalization rates during the isolation test (p = 0.001). the G-1.0 group had the lowest H/L ratio (p = 0.001), but higher plasma tryptophan concentrations and a greater number of birds could reach the observer during the touch test (p = 0.001 and 0.043, respectively). The current results indicate that the synbiotic can be used as a growth promoter to reduce the fear response and stress state of heat-stressed broilers.

A Comparative Transcriptomic Analysis of Human Placental Trophoblasts in Response to Pathogenic and Probiotic Enterococcus faecalis Interaction.

With the ability to cross placental barriers in their hosts, strains of Gram-positive Enterococcus faecalis can exhibit either beneficial or harmful properties. However, the mechanisms underlying these effects have yet to be determined. A comparative transcriptomic analysis of human placental trophoblasts in response to pathogenic or probiotic E. faecalis was performed in order to investigate the molecular basis of different traits. Results indicated that both E. faecalis Symbioflor 1 and V583 could pass through the placental barrier in vitro with similar levels of invasion ability. In total, 2353 (1369 upregulated and 984 downregulated) and 2351 (1233 upregulated and 1118 downregulated) DEGs were identified in Symbioflor 1 and V583, respectively. Furthermore, 1074 (671 upregulated and 403 downregulated) and 1072 (535 upregulated and 537 downregulated) DEGs were only identified in Symbioflor 1 and V583 treatment groups, respectively. KEGG analysis showed that 6 and 9 signaling pathways were associated with interactions between Symbioflor 1 and V583. GO analysis revealed that these DEGs were mainly related to cellular and metabolic processes and biological regulation. However, 28 and 44 DEGs were classified into terms associated with placental and embryonic development in Symbioflor 1 and V583 treatment groups, respectively. Notably, 9 and 25 unique DEGs were identified only in Symbioflor 1 and V583 treatment groups, respectively. A large proportion of transcriptional responses differed when compared between pathogenic and probiotic E. faecalis interaction, and several unique DEGs and signal pathways were identified in the two different groups. These data enhance our understanding of how different traits can be affected by pathogenic and probiotic E. faecalis and the mechanisms underlying these effects.

In vitro evaluation of the antiviral activity and detection of the enterocin coding genes of the probiotic bacterium Enterococcus faecium NM213

In vitro evaluation of the antiviral activity and detection of the enterocin coding genes of the probiotic bacterium Enterococcus faecium NM213

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.

Probiotic Enterococcus faecalis AG5 mitigated high fat diet induced obesity and produced propionic acid stimulated apoptosis in 3T3-L1 pre-adipocyte

AimsProbiotics and their metabolites (SCFA) can regulate energy homeostasis. The present study thus evaluates the synergistic effect of probiotic Enterococcus faecalis AG5 on HFD induced obesity and the role of propionic acid (PA) in apoptosis induction of 3T3-L1 pre-adipocyte. MethodsMale Wistar rats (n = 24) were used to develop an HFD induced obesity model for 24 weeks. The effect of the orally administered probiotic AG5 (18th–24th weeks, 1 × 109 CFU/ml) was evaluated using physiological, biochemical, anthropometry, histopathological and serological analyses. Apoptosis in 3T3-L1 pre-adipocyte was assessed using flow cytometry, protein expression of PPARγ, 5-LOX, NF-κB, p-AKT, caspase 10 and detection of caspase 3/7 by Immunofluorescence confirmed the apoptosis induced by PA. Key findingsProbiotic AG5 significantly reduced body weight, BMI, serum cholesterol, triglycerides (p < 0.05) and improved HDL, insulin and leptin but lowered LDL and VLDL (p > 0.05). An inflammatory response was reduced as evident by TNF-α IHC. AG5 reduced adipocyte hypertrophy and fatty acid accumulation. Flow cytometry confirmed late apoptosis in PA-AG5 and standard PA treated 3T3-L1 cells. 5-LOX inhibition is associated with apoptosis induction, and increased caspase 1p 10 is related to cell death initiation. The study initially showed a low PPARγ activity inhibiting 5-LOX which may relate to adipose apoptosis. Finally caspase 3/7 detection using immunofluorescence proved the role of PA in adipocyte apoptosis. SignificanceThe present study is a novel approach towards obesity mitigation involving adipocyte apoptosis. The role of SCFA in adipocyte apoptosis is very limited which can prove to be novel therapeutic approach in the future.

Field evaluation of the influence of garlic extract and probiotic cultures on sows and growing pigs

The present study aimed to evaluate the influence of a garlic extract and probiotic strains, Enterococcus faecium, Lactobacillus rhamnosus and Lactobacillus fermentum, on sows and their offspring. A total of 24 sows and the 301 piglets they farrowed were used in this trial. The animals (sows and piglets) were assigned to 3 groups: group A, supplemented with garlic extract; group B, supplemented with probiotics; and the control group C, without supplementation. The trial lasted from the 80th day of gestation to the weaning day for sows. For piglets, results were analyzed during two periods, from the 8th day to the 56th or the 147th day of life, in order to determine the more desirable duration of supplementation. Morphological and biochemical blood parameters, as well as body weight and piglet losses, were recorded. At weaning, the piglets supplemented with probiotics were heavier than the rest of experimental population (P ≤ 0.01). On the day of sale for slaughter, the pigs from the control group were on average 15 kg lighter (P ≤ 0.01) than those from both supplemented groups. Differences in blood parameters were also noted between the groups. Our findings suggest that the use of a garlic extract and probiotic bacteria may help improve pig performance at various stages of the production cycle.

Effect of fermented sausages with probiotic Enterococcus faecium CRL 183 on gut microbiota using dynamic colonic model

Considering the importance of nutrition in maintaining health conditions, this study is aimed at evaluating the effect of the consumption of a potentially probiotic sausage, with reduction of curing salts and fat, fermented with Enterococcus faecium CRL 183 into the gut microbiota, using the simulator of the human microbial ecosystem (SHIME®). The SHIME® experiment was performed using the probiotic sausage for two weeks. The gut microbiota composition was monitored weekly in all colon vessels by plate counts and denaturing gradient gel electrophoresis (PCR-DGGE). Samples of the colonic contents were evaluated microbiologically by plate counts and denaturing gradient gel electrophoresis (PCR-DGGE) and chemically by the production of SCFA (acetate, propionate and butyrate) and ammonium ions (NH+4). It was observed that the consumption of the potentially probiotic sausage can result in a reduced count of Lactobacillus spp. (colon vessels 3, 4 and 5), Bacteroides spp. (colon vessel 5) and Enterobacteriaceae (colon vessels 4 and 5) and increased levels of ammonium ions. The butyric and propionic acids increased (P < 0.05) while acetic acid decreased (P < 0.05) in treatment period. The PCR-DGGE analysis suggested that probiotic sausage increased the microbiota diversity during the treatment phase. Overall, the findings showed that potentially probiotic sausage represents a healthier alternative of fermented meat, with a positive effect on the microbial diversity and metabolite production.

Exopolysaccharide produced by potential probiotic Enterococcus faecium MS79: Characterization, bioactivities and rheological properties influenced by salt and pH

Microbial exopolysaccharide (EPS) is of interest due to their applications in food processing and potential health benefits following consumption. The objectives of this study were to: 1) isolate, purify and characterize EPS produced by Enterococcus faecium MS79 (EPS-MS79); 2) investigate bioactive properties of EPS-MS79; and 3) investigate the rheological properties of the EPS-MS79 as influenced by salt type and pH level. The average molecular weight of the EPS-MS79 was 8.3 × 105 Da and it is composed of the three monosaccharides arabinose, mannose, and glucose in a molar ratio of 0.8:1.7:11.3. Scavenging activities of EPS-MS79 were 76% and 85% as measured by DPPH and 26% and 44% as measured by ABTS. The antiproliferative activities against colon (72% and 77%) and breast (43% and 56%) cancer cell lines. EPS-MS79 inhibited the growth of bacterial pathogens by up to 3 logs compared to the control. All EPS-MS79 solutions displayed shear thinning behavior. The storage and loss moduli of all EPS-MS79 solutions increased alongside with the frequency increase. Salt type and pH level impacted on the rheological properties of EPS-MS79. The EPS-MS79 could be a potent prebiotic promoting health and rheological properties of foods.