Cholesterol Assimilation Research Articles

A potential probiotic Enterococcus faecium isolated from camel rumen, fatty acids biotransformation, antilisteria activity and safety assessment

The probiotic features of a new Enterococcus faecium strain isolated from camel rumen were investigated. The isolate identified as E. faecium 96B4 exhibited notable tolerance to gastrointestinal conditions, with high cell viability at pH 1.5 after 3 h (about 7.48 log10 cfu mL−1). Besides complete inhibition of Listeria monocytogenes growth, E. faecium 96B4 showed high hydrophobicity, auto- and co-aggregations abilities. It showed 76.33% cholesterol assimilation ability and boosted the extracellular unsaturated fatty acid content up to 25.53% in the presence of soybean oil as a substrate. Moreover, 96.17 ± 3.52 μg mL−1 conjugated linoleic acid production and 2.56 fold increase in oleic acid content were highlighted. Significant diacetyl production ability was also demonstrated. E. faecium 96B4 passed the safety assessment successfully. The association between the microbiome and host health makes the camel, as a unique animal, a rich source of beneficial microorganism candidates for designing future foods.

The Expression Levels of Genes Responsible for the Enzymatic Activity of Bile Salt Hydrolase (BSH) and the Relationship of Cholesterol Assimilation in L. plantarum and L. paracasei.

The bile salt hydrolase (BSH) activity is responsible for the cholesterol-lowering effect of the probiotic strains. The present study aimed to investigate the relationship between bsh gene-expression (GE) levels responsible for the BSH activity and the parameters of bile salt resistance of different Lactobacillaceae species. Accordingly, 11 Lactobacillaceae family strains with high cholesterol assimilation ratio (49.21-68.22%) determined by the o-phthalaldehyde method selected from 46 Lactobacillaceaespecies was evaluated for their features including acid tolerance, bile tolerance, and BSH activity. All tested strains survived at pH 2 medium and 0.3% (w/v) bile salt and showed positive BSH activity for glycocholic acid (GCA) and taurocholic acid (TCA). BSH gene expression was performed to provide clear information and to identify the key genes responsible for BSH activity. bsh3 genes were found highest GE level (P < 0.05) in Lactiplantibacillus plantarum and Lacticaseibacillus paracasei strains. The results showed that high cholesterol assimilation ratio were closely correlated with BSH activity and the parameters of bile salt resistance. The results of this study will support the development of a new approach based on phenotypic and genetic analysis to determine the bile salt parameters. The study will be useful for the selection of Lactobacillus strains with high bile salt resistance.

Evaluating the Probiotic Potential of Lactic Acid Bacteria Implicated in Natural Fermentation of Table Olives, cv. Cobrançosa.

The probiotic features of Lactiplantibacillus (L.) pentosus and L. paraplantarum strains, endogenous in Cobrançosa table olives from northeast Portugal, were assessed in terms of functional properties and health benefits. Fourteen lactic acid bacteria strains were compared with Lacticaseibacillus casei from a commercial brand of probiotic yoghurt and L. pentosus B281 from Greek probiotic table olives, in attempts to select strains with higher probiotic performances than those references. For functional properties, the i53 and i106 strains, respectively, exhibited: 22.2 ± 2.2% and 23.0 ± 2.2% for Caco-2 cell adhesion capacity; 21.6 ± 7.8% and 21.5 ± 1.4% for hydrophobicity; 93.0 ± 3.0% and 88.5 ± 4.5% for autoaggregation ability by 24 h of incubation; and ability to co-aggregate with selected pathogens-from 29 to 40% to Gram+ (e.g., Staphylococcus aureus ATCC 25923 and Enterococcus faecalis ATCC 29212); and from 16 to 44% for Gram- (e.g., Escherichia coli ATCC 25922 and Salmonella enteritidis ATCC 25928). The strains proved to be resistant (i.e., halo zone ≤14 mm) to some antibiotics (e.g., vancomycin, ofloxacin, and streptomycin), but susceptible (i.e., halo zone ≥ 20 mm) to others (e.g., ampicillin and cephalothin). The strains exhibited health-beneficial enzymatic activity (such as acid phosphatase and naphthol-AS-BI-phosphohydrolase), but not health-harmful enzymatic activity (such as β-glucuronidase and N-acetyl-β-glucosaminidase). Additionally, the antioxidant activity and cholesterol assimilation features, respectively, of the strains were 19.6 ± 2.8% and 77.5 ± 0.5% for i53, and 19.6 ± 1.8% and 72.2 ± 0.9% for i106. This study indicated that the addition of L. pentosus strains i53 and/or i106 to Cobrançosa table olives is likely to enhance the added value of the final product, in view of the associated potential benefits upon human health.

Evaluation of the Probiotic Properties of Lacticaseibacillus casei 431® Isolated from Food for Special Medical Purposes§.

Increasing awareness of the importance of nutrition in health promotion and disease prevention has driven to the development of foods for special medical purposes (FSMPs). In this study, the probiotic strain Lacticaseibacillus paracasei ssp. paracasei (Lacticaseibacillus casei 431®) was incorporated into FSMPs to develop an innovative product. The aim was to investigate the influence of the FSMP matrix on the specific probiotic properties of L. casei 431® in vitro. A series of in vitro experiments were performed as part of the probiotic approach. After evaluation of antibiotic susceptibility profiles, functional properties such as survival under simulated gastrointestinal tract (GIT) conditions, bile salt deconjugation activities, cholesterol assimilation, antagonistic activity against spoilage bacteria and adhesion to Caco-2 cell line monolayers and extracellular matrix proteins were investigated. The L. casei 431® strain, both the lyophilised strain and the strain isolated from the FSMP matrix, effectively survived the simulated adverse gastrointestinal conditions without significant effects of the food matrix. The effect of the FSMP matrix on the deconjugation activity of the bile salts of L. casei 431® was minimal; however, cholesterol assimilation was increased by 16.4 %. L. casei 431® had antibacterial activity against related lactic acid bacteria regardless of whether it was used in FSMPs or not. Conversely, the probiotic strain isolated from FSMP matrix had significantly higher inhibitory activity against six potential pathogens than the lyophilised culture. The autoaggregation ability of the L. casei 431® cells was not affected by the FSMP matrix. The adhesion of L. casei 431® bacterial cells to the extracellular matrix proteins was reduced after treatment with proteinase K, with the highest adhesion observed to laminin. The adhesion of L. casei 431® reduced the binding of E. coli 3014 by 1.81 log units and the binding of S. Typhimurium FP1 to Caco-2 cell lines by 1.85 log units, suggesting the potential for competitive exclusion of these pathogens. The results support the positive effect of the FSMP matrix on the specific probiotic properties of L. casei 431®, such as antibacterial activity, bile salt deconjugation and cholesterol assimilation, while the incorporation of this probiotic strain adds functional value to the FSMPs. The synergistic effect achieved by the joint application of L. casei 431® and innovative FSMP matrix contributed to the development of the novel formulation of an improved functional food product with added value.

Assessment of Genomic and Metabolic Characteristics of Cholesterol-Reducing and GABA Producer Limosilactobacillus fermentum AGA52 Isolated from Lactic Acid Fermented Shalgam Based on "In Silico" and "In Vitro" Approaches.

This study aimed to characterize the genomic and metabolic properties of a novel Lb. fermentum strain AGA52 which was isolated from a lactic acid fermented beverage called "shalgam." The genome size of AGA52 was 2,001,184bp, which is predicted to carry 2024 genes, including 50 tRNAs, 3 rRNAs, 3 ncRNAs, 15 CRISPR repeats, 14 CRISPR spacers, and 1 CRISPR array. The genome has a GC content of 51.82% including 95 predicted pseudogenes, 56 complete or partial transposases, and 2 intact prophages. The similarity of the clusters of orthologous groups (COG) was analyzed by comparison with the other Lb. fermentum strains. The detected resistome on the genome of AGA52 was found to be intrinsic originated. Besides, it has been determined that AGA52 has an obligate heterofermentative carbohydrate metabolism due to the absence of the 1-phosphofructokinase (pfK) enzyme. Furthermore, the strain is found to have a better antioxidant capacity and to be tolerant to gastrointestinal simulated conditions. It was also observed that the AGA52 has antimicrobial activity against Yersinia enterocolitica ATCC9610, Bacillus cereus ATCC33019, Salmonella enterica sv. Typhimurium, Escherichia coli O157:h7 ATCC43897, Listeria monocytogenes ATCC7644, Klebsiella pneumoniae ATCC13883, and Proteus vulgaris ATCC8427. Additionally, AGA52 exhibited 42.74 ± 4.82% adherence to HT29 cells. Cholesterol assimilation (33.9 ± 0.005%) and GABA production capacities were also confirmed by "in silico" and "in vitro." Overall, the investigation of genomic and metabolic features of the AGA52 revealed that is a potential psychobiotic and probiotic dietary supplement candidate and can bring functional benefits to the host.

Characterization of Short-chain fatty acid-producing and cholesterol assimilation potential probiotic Lactic acid bacteria from Chinese fermented rice

Probiotic strains are phrased as beneficial for life because this bacterial association has substantial effects on human and animal health when administrated orally, among which bio-active metabolite short-chain fatty acids production and cholesterol assimilation are most significant. All the short-chain fatty acids, mainly acetate, propionate, and butyrate reveal valuable effects on human health. In this research, the ability of 9 the potential lactic acid bacteria food-isolated probiotics was investigated to produce bio-active metabolite short-chain fatty acids directly or indirectly, and cholesterol-lowering ability through various mechanisms. Their 16S rRNA gene sequences were analyzed. short-chain fatty acids were quantified by gas chromatography, while cholesterol assimilation was measured using the o-phthalaldehyde method with slight modifications. All isolated strains through this study have the ability to optimize cholesterol content in the culture broth, short-chain fatty acid production, and survival in extreme alkaline and acidic conditions. The highest resistance to bile salt concentrations, γ-hemolysis, and strong antibacterial activity against pathogenic indicators, especially Escherichia coliATCC25922. The highest Lipid accumulation inhibitory effect was noticed in Lentilactobacillus hilgardii LMG 07934 as a potential probiotic isolate on the basis of cholesterol assimilation capacity (i.e., 91.4%), along with the ability to produce abundant i.e., butyric acid, propionic acid, and acetic acid, followed by Lacticaseibacillus paracasei ATCC 334 which has 85.3% cholesterol assimilation ability. These findings demonstrated that these isolated strains from Chinese fermented rice can be used as a potential probiotic and alternate therapeutic supplement because of their ability to produce short-chain fatty acids and their cholesterol-lowering ability.

Multifunctional potentials of lactic acid bacterial isolates from Turkish traditional fermented foods.

This study aimed to isolate LAB strains with antimicrobial activity that can be used as bio-protective agents, from yogurt, pickle, and cheese samples, which are Turkish traditional fermented foods, and to reveal the potential probiotic properties of these isolates with antimicrobial activity. In addition, it was aimed for the first time to examine the potential and uniqueness of these domestic and local traditional fermented foods. In the study, a total of 682 LAB isolates from Turkish traditional fermented samples were isolated. Thirteen isolates with both high and antifungal activities were selected; four Lactiplantibacillus pentosus strains, six Lactiplantibacillus plantarum strains, and one each Enterococcus lactis, Enterococcus durans, and Enterococcus faecalis strains. Ten isolates of them were suggested as potential probiotics (excluding 1Y2-1, 1Y7-3, and 3Y12-4). It was thought that the L. pentosus 2Y7-1, which was slow to produce acid, could be used as a co-culture in fermented products with a long ripening stage, supported by enriching the product of its milk coagulation, β-galactosidase, proteolytic, and cholesterol assimilation abilities in terms of bioactive peptide and prebiotics. Another strain, E. faecalis 3Y4-3, which was high proteolytic activity, was suggested as a co-culture in fermented products to improve both organoleptic properties and increase bioactive peptides. On the other hand, among non-probiotics, L. plantarum 3Y12-4 can be used as a starter culture in fermented milk products in order to evaluate its milk coagulation, cholesterol assimilation, and proteolytic abilities. As a result, these three strains can be used in many areas, such as fermented and/or functional food production and enzyme production. In addition, this has been the first study to examine the potential and uniqueness of Turkish domestic and local traditional fermented foods in Samsun, Turkey, together, in terms of both antimicrobial and probiotic LAB isolate (multifunctional) potential and diversity.

Salutary Attributes of Probiotic Human Gut Lactobacilli for gut health.

Lactobacilli are GRAS organisms and are important members of gut microbiota. They have been strongly recommended as probiotics because of many benefits provided by them to overall human health. Human gut lactobacilli with salutary properties can provide additional advantages. Limosilactobacillus fermentumL1 MW600457, Limosilactobacillus fermentumL3 MW600480, Limosilactobacillus fermentumL4 MW600464, Limosilactobacillus fermentumL5 MW600493, Limosilactobacillus fermentumL6 MW600495, Limosilactobacillus fermentumL7 MW600496, Limosilactobacillus fermentumL8 MW485761, Lactiplantibacillus plantarumL9 MW485746 and Ligilactobacillus salivariusL10 MW600498 with in vitro probiotic properties were explored for salutogenic characteristics. Salutary properties like β-galactosidase activity, anthelminthic property assay, anti-inflammatory assay, anti-diabetic study, cholesterol assimilation assay and biofilm assay were performed. All the isolates were positive for β-galactosidase activity. The anthelminthic property with minimum paralysis time and death time between 16-25min was shown by Limosilactobacillus fermentumL8 MW485761. Anti-inflammatory activity with bovine serum albumin was maximum at 200μg ml-1 concentration. α-amylase inhibitory activity was maximum for Limosilactobacillus fermentumL8 MW485761 (51%). Maximum cholesterol assimilation was reported for Limosilactobacillus fermentumL6 MW600495 (76%). Strong biofilm forming ability for all isolates improved the colonization and stability as probiotics. Human gut lactobacilli with salutary properties can make an efficient probiotic.

PROBIOTIC CHARACTERIZATION OF SACCHAROMYCES CEREVISIAE Y196 AND Y197 ISOLATED FROM RICE CHHANG- A FERMENTED BEVERAGE OF LAHAUL SPITI

Fermented foods and beverages are rich source of probiotic microorganisms and yeasts are the most common microorganisms found in these fermented products. Fermented beverages are less explored for probiotics, therefore; the present study was focused on isolation, screening and characterization of probiotic yeast from rice Chhang, a fermented beverage of Lahaul Spiti. Ten yeast isolates were initially isolated from rice Chhang and out of which two isolates Saccharomyces cerevisiae Y196 and Saccharomyces cerevisiae Y197 were selected based on their antimicrobial activity. These two yeast isolates were examined for various probiotic properties such as survival at low pH, bile tolerance, microbial adhesion to hydrocarbons, cholesterol assimilation, exopolysaccharide production and haemolytic activity. Both isolates showed antimicrobial activity against all the tested pathogens used in this study, also showed high acid tolerance at pH 2 and pH 3 and can withstand high bile concentration as compared to control. Percentage of hydrophobicity was found to be 57 % and 41 % in Saccharomyces cerevisiae Y196 and Saccharomyces cerevisiae Y197, respectively. Both isolates were able to show high cholesterol assimilation with a maximum of 90 % in case of Saccharomyces cerevisiae Y197 with taurocholate. They were also found to be positive for exopolysaccharide production and showed no haemolytic activity. Hence, keeping in view the results of present study, these isolates can be proposed for preparation of various probiotic food products.

The Mechanism of Probiotics in Preventing the Risk of Hypercholesterolemia

Probiotics are beneficial live microorganisms because they provide health benefits to the host after being consumed in sufficient amounts, primarily by increasing the proliferation of native digestive microflora. Hypercholesterolemia is a condition in which cholesterol serum concentration is elevated significantly (exceeds 200 mg/dL). Hypercholesterolemia is characterized by high plasma levels of low-density lipoprotein (LDL) and low plasma concentration of high-density lipoprotein (HDL). This review aims to examine the mechanism of action of probiotics in preventing the risk of hypercholesterolemia and reducing LDL cholesterol levels in the human body. The mechanism of cholesterol-lowering effect of probiotic bacteria can be classified into four basic principles such as 1) the conversion of cholesterol to coprostanol, 2) the deconjugation of cholesterol by probiotics’ bile salt hydrolase enzyme, 3) the alteration of protein expression related to cholesterol synthesis, and 4) the production of Short Chain Fatty Acids (SCFA) by probiotics. Cholesterol converted to coprostanol is not absorbed by the small intestine and is directly excreted with the feces. The deconjugated cholesterol is easily precipitated and thus not absorbed by the small intestine. It also induces the assimilation of cholesterol into the probiotic membrane. Finally, the SCFAs, produced by probiotics fermentation within host digestive tract, can upregulate the synthesis of HDL apolipoprotein.

Distribution, cholesterol-lowering and immunomodulation effects of lactic acid bacteria from fermented mussel (Hoi-dong)

Forty-eight lactic acid bacteria (LAB) isolated from fermented mussels in Thailand were evaluated for their probiotic properties, bile salt hydrolase (BSH), cholesterol assimilation and immunomodulatory effects. They were identified as Companilactobacillus formosensis (Group I, 10 isolates), Lentilactobacillus buchneri (Group II, 8 isolates), Lactiplantibacillus plantarum subsp. plantarum (Group III, 16 isolates), Lacticaseibacillus rhamnosus (Group IV, 1 isolate), Pediococcus pentosaceus (Group V, 5 isolates) and P. acidilactici (Group V, 1 isolate), Enterococcus thailandicus (Group VI, 2 isolates), En. hirae (Group VII, 1 isolate), En. durans (Group VI, 1 isolate), Lactococcus lactis subsp. lactis (Group VII, 1 isolate), Lc. lactis subsp. hordinae (Group VII, 1 isolate), and Leuconostoc lactis (Group VIII, 1 isolate), based on their phenotypic and genetic characteristics. Seven isolates, L. plantarum subsp. plantarum LM6-1, LM6-2, LM7-2-2B, LM12-1, LM14-1, LM15-1P and LM15-2 expressed bile salt hydrolase activity. All isolates assimilated cholesterol ranging from 20.73 to 79.40%. BSH-producing isolates were tolerant to acidic and bile conditions and showed the adhesion ability to Caco-2 cells. The BSH-producing and selected isolates showed the immunomodulatory effects to stimulate interleukin-12 (IL-12), interferon-gamma (IFN-γ), human beta defensin-2 (hBD-2) and nitric oxide (NO) production at various levels. Therefore, these results indicated that the isolates meet the standard probiotic criteria and beneficial effects.

Enterococcus faecium GEFA01 alleviates hypercholesterolemia by promoting reverse cholesterol transportation via modulating the gut microbiota-SCFA axis.

This study aimed to identify cholesterol-lowering commensal strains from healthy lean individuals and to evaluate the cholesterol-lowering capacity of Enterococcus faecium GEFA01 in mice fed a high-cholesterol and high-fat diet. E. faecium GEFA01 was isolated from the feces of a healthy lean individual in a selective basal salt medium supplemented with cholesterol. E. faecium GEFA01 exhibited a cholesterol removal rate (CRR) of 46.13% by coprecipitation, assimilation, and degradation of cholesterol. Moreover, E. faecium GEFA01 significantly decreased the body weight of mice and the levels of serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), hepatic TC, triglycerides (TG), and LDL-C, and increased serum high-density lipoprotein cholesterol (HDL-C) levels in mice fed a high-cholesterol diet compared with the HCD group. We also observed that E. faecium GEFA01 significantly downregulated the gene expression of HMG-CoA reductase (Hmgcr), Srebp-1c, Fxr, Shp, and Fgf 15, upregulated the gene expression of low-density lipoprotein receptor (Ldlr), Abcg5/8, Abca1, cholesterol 7 alpha-hydroxylase (Cyp7a1), and Lxr in the liver of mice in relative to the HCD group, markedly increased the relative abundance of Lactobacillus, Akkermansia, Bifidobacterium, and Roseburia, and decreased the abundance of Helicobacter in the feces. Collectively, we confirmed that E. faecium GEFA01 exhibited cholesterol-lowering effects in mice fed a high-cholesterol diet, which was achieved through assimilation, coprecipitation, and degradation of cholesterol, and through modulation of the gut microbiota short-chain fatty acid (SCFA) axis that promoted reverse cholesterol transport and bile acid excretion. Our study demonstrated that E. faecium GEFA01 may be used as a probiotic candidate to lower cholesterol levels in the future.

Lactobacillus spp. isolated from prebiotic-derived raw goat milk: probiotic characteristics, cholesterol assimilation and folate production.

This work provides to evaluate cholesterol assimilation and folic acid production by determining the probiotic properties of Lactobacillus spp. from raw goat milk with prebiotic properties. We isolated Lactobacilli from goat milk and identified API 50, CHL, and 16sRNA. Probiotic properties were determined according to bile salt and acidic tolerance, hydrophobicity, hemolytic activity, antibiotic sensitivity, antagonistic effect, and exopolysaccharide production. In addition, the cholesterol assimilation and folate production of cultures were determined. L. plantarum GM-12 and L. plantarum GM-15 showed the highest folate production and the highest cholesterol assimilation.These two strains are strong candidates for use as potential probiotics and starter cultures.

Evaluation of probiotic potential of autochthonous lactobacilli strains isolated from Zabuli yellow kashk, an Iranian dairy product.

The aim of this study was to evaluate the probiotic potential and anti-biofilm activity of five lactobacilli strains which isolated and identified from an Iranian product. Five lactobacilli strains, which were isolated from Zabuli yellow kashk, were evaluated for the presence of probiotic properties, such as resistance to low pH, resistance to simulated gastrointestinal conditions, bile salt tolerance, hydrophobicity, and auto- and co-aggregation. In addition, antimicrobial susceptibility, adherence to Caco-2 cells (human colon cancer cell line), anti-adhesion activity, ability against biofilm formation and biofilm degradation of mentioned strains against Pseudomonas aeruginosa PTCC 1707 were assessed. All the strains tested showed acceptable characteristics, but Lactiplantibacillus plantarum TW57-4 appeared of particular interest. Some probiotic properties of this strain were similar and in some cases higher than the commercial probiotic strain Lacticaseibacillus rhamnosus GG (standard sample). Cholesterol assimilation and radical-scavenging activity of Lpb. plantarum TW57-4 were 70.2% and 62.3%, respectively. The adhesion degree of Lpb. plantarum TW57-4 was 10.6%. Applying competition and inhibition assay, this strain showed 55.3% and 62.3% of competition and inhibition activity in adhesion of P. aeruginosa PTCC 1707 to the intestinal cells, respectively. According to the obtained results, it can be concluded that Lpb. plantarum TW57-4 strain can be used as a promising candidate for in-vivo studies with the aim of developing new probiotic starter cultures. The present study furthers our understanding of lactobacilli strains behaviour after consumption to establish their beneficial effects.

Deciphering Beneficial Health Promoting Properties of Lactic Acid Bacteria Isolated From Infant Gut Microbiome

Aim and Objective: To evaluate the probiotic potenties along with safety of probiotics within infant faecal origin. Methods and Results: The present study isolates lactic acid bacteria from infant feces were characterized and identified by 16S rRNA sequencing. On evaluation of in vitro probiotic capability, the test isolates exhibited in vitro probiotic potency, antagonistic property and better cell surface properties. The Bile salt hydrolase activity, cholesterol assimilation, antioxidant activity and angiotensin converting enzyme inhibitory action of the test isolates proves their health conferring potency. Standard protocols were used for the estimation of lymphocyte proliferation rate, phagocytosis rate, cytokine response and faecal enzyme levels. Animal experiment using different doses of L. plantarum (MBTU-S1B) substantiated its immunotolerance against inhibitory effects of supraoptimal concentration of both T and B cell mitogens with significant reduction on β-Glucosidase and Nitroreductase activity with p&lt; 0.05, while contrasting effect was noticed for β-Glucuronidase. Findings of the in vivo study proved higher dose of probiotic would not provide any additional benefit on host body. Conclusions: The test isolates possesed in vitro probiotic potency and the in vivo study with MBTU- S1B proved their applicability in nutraceutical and pharmaceutical industries. Significance and Impact of Study: To conflict disease outbreaks by improving general immunity via probiotics.

In vitro evaluation of probiotic potential and enzymatic profiling of Pichia kudriavzevii Y33 isolated from traditional home-made mango pickle

BackgroundFermented foods are the results of metabolic activities of various microorganisms. People have traditionally known how to culture desirable microorganisms, primarily lactic acid bacteria, yeasts, and filamentous molds, for the manufacture of edible foods. Yeast isolated from home-made mango pickle from Hamirpur, Himachal Pradesh, was assessed for probiotic properties and their enzymatic profiling. ResultsFour yeast isolates were isolated out of which P. kudriavzevii Y33 was selected on the basis of high acid tolerance as well as broadest antimicrobial activity. The selected isolate was observed to have high acid tolerance at pH 2 and show strong antimicrobial activity against all the pathogens examined. P. kudriavzevii Y33 can also withstand high bile concentration and showed high viability index, i.e., 95% at concentration of 2% of bile. The isolate was able to demonstrate high cholesterol assimilation in medium containing ox bile and taurocholate, at 88.58 and 86.83%, respectively. The autoaggregation ability of isolate increases with increasing the time of incubation and showed 87% of autoaggregation after 24 h of incubation. P. kudriavzevii Y33 exhibited resistance towards different antibiotics, found to be positive for exopolysaccharide production and showed no hemolytic activity. The isolate was observed to produce several enzymes such as β-galactosidase, protease, amylase, phytase, and lipase. ConclusionsThe results of the current study revealed that P. kudriavzevii Y33 has various beneficial qualities that suggest it could be used as probiotics. Enzymes produced by yeast isolate help in improving flavor and mineral availability in the fermented products.

Isolation and characterisation of Pulsatilla Radix-utilising bacteria Pediococcus pentosaceus PR-1 from human faeces.

Although probiotics have been isolated from different sources, few were isolated from traditional Chinese medicine. The current study firstly isolates Pulsatilla Radix-utilising Pediococcus pentosaceus PR-1 from human faeces. Subsequently, the tolerance of PR-1 to low pH, bile salts, simulated gastric juice and succus entericus, antioxidant activity, antimicrobial activity, cholesterol assimilation and antibiotics susceptibility were investigated. After 2h of incubation at pH 2.0, over 80% of PR-1 survived. The cell viability of PR-1 at 2h under 0.1% bile salt condition was 99.2%. The survival rate of PR-1 in gastric juice and succus entericus was 64.48% and 81.86%, respectively. Cell-free supernatant of PR-1 culture also showed antimicrobial activity against Escherichia coli, Staphylococcus aureus and Salmonella typhimurium. Besides, antioxidant activity of PR-1 CFS was significantly greater than cell pellet. PR-1 was shown to be resistant to kanamycin, streptomycin, vancomycin and norfloxacin and was able to lower the cholesterol level to 72.5±1.5%. In addition, PR-1 displayed γ-haemolysis and was non-pathogenic.

Characterization of autochthonous lactobacilli from goat dairy products with probiotic potential for metabolic diseases

The present study aimed to design functional fermented goat milk with probiotic potential for metabolic diseases. Thereby, autochthonous lactobacilli from goat dairy products that target improving the inflammatory, lipid, and glycemic profile were characterized. We designed fermented goat milk using Lactobacillus delbrueckii subsp. indicus CRL1447 as starter strain, supplemented with different probiotic consortia formed by Limosilactobacillus fermentum CRL1446, Lactiplantibacillus paraplantarum CRL1449, and CRL1472 strains. These lactobacilli were selected for their positive effects on inhibition of α-glucosidase, bile salts hydrolase activity, cholesterol assimilation, and decreased triglyceride percentage in Caenorhabditis elegans. Furthermore, the lactobacilli oral administration to obese mice caused a significant decrease in body weight gain and ameliorated hyperglycemia and hyperlipemia. These results reveal the potential of this goat dairy product as a functional food to prevent obesity and related pathologies. Goat milk-derived products stand out for their marketing potential. Hence, fermented goat milk incorporating novel probiotics represents a group of food products with broad prospects by their promising nutritive and therapeutic properties for metabolic diseases. The goat dairy product designed in this study could be used in the prevention of dyslipidemia and hyperglycemia in obese people.

Bile salt hydrolase and cholesterol assimilation potential of lactobacilli fromNigerian fermented foods and human sources

Bile salt hydrolases (BSH) results in microbes releasing enzymes conferring physiological functions for both the producing organisms and the host and subsequently lower serum cholesterol. In recent years, these attributes have generated interest towards screening of lactic acid bacteria (LAB) for functional food production. In this present study, lactobacilli (n=34) (13 L. casei, 6 L. paracasei, 11 L. brevis, 3 L. pentosus and 1 L. plantarum) from varied sources viz. Nigerian fermented foods and human specimens were screened qualitatively and quantitatively for BSH activity using the direct plate assay and cell free extracts (CFE) of overnight cultures. To establish a correlation between BSH and cholesterol reduction, 12 isolates displaying high precipitation over bile salt agar plates, and high BSH enzyme activity in CFE were selected for in vitro cholesterol reduction assay. Results showed that 58% of the isolates from humans, and 36% from fermented food origin showed larger zones of precipitation. The highest BSH activity was exhibited by Lactobacillus casei GR4 (76mM) after 10 min of incubation, an isolate from fermented food. Isolates from human specimens expressed higher enzyme activity compared to those from fermented foods with significant increase (P&lt;0.05) after 30 min incubation. Cholesterol level was variable as L. pentosus 8ST5 and 8ST7 had 2 and 4% cholesterol levels in the medium; while L. casei and paracasei group (Lactobacillus casei group, LCG) 3MB4, 8BM6, BK4 and L. brevis GR29 had 7%/ml each proving their application in food formulation, reduction of hypercholesterolemia, and as prophylaxis for controlling serum cholesterol levels.

Mimicking natural cholesterol assimilation to elevate the oral delivery of liraglutide for type II diabetes therapy

Glucagon-like peptide-1 receptor agonists (GLP-1 RA) are a series of polypeptides broadly applied in the long-term treatment of type Ⅱ diabetes. However, administration of GLP-RA is mainly through repetitive subcutaneous injection, which may seriously decrease the compliance and safety. Herein, a bio-inspired oral delivery system was designed to enhance the oral absorption of liraglutide (Lira), a kind of GLP-1 RA, by mimicking the natural cholesterol assimilation. 25-hydroxycholesterol (25HC), a cholesterol derivative, was modified on the surfaced of Lira-loaded PLGA nanoparticles (Lira 25HC NPs) and functioned as a “top-down” actuator to facilitate unidirectional transcytosis across the intestinal epithelium. After oral delivery, Lira 25HC NPs displayed improved therapeutic effect as compared with oral free Lira on type Ⅱ diabetes db/db mice, as evidenced by multiple relieved diabetic symptoms including the enhanced glucose tolerance, repressed weight growth, improved liver glucose metabolism, decreased fasting blood glucose, HbA1c, serum lipid, and increased β cells activity. Surprisingly, the fasting blood glucose, liver glucose metabolism, and HbA1c of oral Lira-loaded 25HC NPs were comparable to subcutaneous injection of free Lira. Further mechanisms revealed that 25HC ligand could mediate the nanoparticles to mimic natural cholesterol absorption by exerting high affinity towards apical Niemann-Pick C1 Like 1 (NPC1L1) and then basolateral ATP binding cassette transporter A1 (ABCA1) overexpressed on the opposite side of intestinal epithelium. This cholesterol assimilation-mimicking strategy achieve the unidirectional transport across the intestinal epithelium, thus improving the oral absorption of liraglutide. In general, this study established a cholesterol simulated platform and provide promising insight for the oral delivery of GLP-1 RA.