Design of novel urogenital pharmabiotic formulations containing lactobacilli,salivaricin CRL 1328 and non-microbial compounds with different functionalities

Context: The administration of pharmabiotics is a promising alternative to antimicrobial drugs for the treatment and/or prevention of female urogenital infections.

Objective: To design pharmabiotic formulations including bioactive ingredients of microbial origin combined with non-microbial substances and then to evaluate the stability of the combinations during freeze-drying and storage.

Materials and methods: Different formulations including Lactobacillus gasseri CRL 1263, Lactobacillus salivarius CRL 1328, salivaricin CRL 1328 (a bacteriocin) and non-microbial compounds (lactose, inulin and ascorbic acid) were assayed, and the ingredients were freeze-dried together or separately. The formulations were stored in gelatin capsules at 4?°C for 360?d.

Results: The viability of lactobacilli was affected to different extents depending on the strains and on the formulations assayed. L. salivarius and ascorbic acid were successfully combined only after the freeze-drying process. Salivaricin activity was not detected in formulations containing L. gasseri. However, when combined with ascorbic acid, lactose, inulin or L. salivarius, the bacteriocin maintained its activity for 360?d. The selected microorganisms proved to be compatible for their inclusion in multi-strain formulations together with lactose, inulin and ascorbic acid. Salivaricin could be included only in a L. salivarius CRL 1328 single-strain formulation together with non-microbial substances.

Conclusions: This study provides new insights into the design of urogenital pharmabiotics combining beneficial lactobacilli, salivaricin CRL 1328 and compounds with different functionalities.     

A Newly Developed Synbiotic Yogurt Prevents Diabetes by Improving the Microbiome–Intestine–Pancreas Axis

The prevalence of type 2 diabetes mellitus (T2D) is increasing worldwide, and there are no long-term preventive strategies to stop this growth. Emerging research shows that perturbations in the gut microbiome significantly contribute to the development of T2D, while microbiome modulators may be beneficial for T2D prevention. However, microbiome modulators that are effective, safe, affordable, and able to be administered daily are not yet available. Based on our previous pro- and prebiotic studies, we developed a novel synbiotic yogurt comprised of human-origin probiotics and plant-based prebiotics and investigated its impact on diet- and streptozotocin-induced T2D in mice. We compared the effects of our synbiotic yogurt to those of a commercially available yogurt (control yogurt). Interestingly, we found that the feeding of the synbiotic yogurt significantly reduced the development of hyperglycemia (diabetes) in response to high-fat diet feeding and streptozotocin compared to milk-fed controls. Surprisingly, the control yogurt exacerbated diabetes progression. Synbiotic yogurt beneficially modulated the gut microbiota composition compared to milk, while the control yogurt negatively modulated it by significantly increasing the abundance of detrimental bacteria such as Proteobacteria and Enterobacteriaceae. In addition, the synbiotic yogurt protected pancreatic islet morphology compared to the milk control, while the control yogurt demonstrated worse effects on islets. These results suggest that our newly developed synbiotic yogurt protects against diabetes in mice and can be used as a therapeutic to prevent diabetes progression.     

乳品中活性益生菌选择性检测方法的研究

研究了含活性益生菌乳品中嗜热乳链球菌、保加利亚乳杆菌、嗜酸乳杆菌、双歧杆菌和干酪乳杆菌的选择性计数方法。结果表明：ST—琼脂培养基适合嗜热乳链球菌的计数；LC—琼脂培养基适合干酪乳杆菌的计数；MRS—NNLP琼脂培养基适合双歧杆菌的计数；MRS—pH5．2琼脂培养基适合保加利亚乳杆菌的计数；MRS—柳醇琼脂或MRS—山梨醇琼脂可用于嗜酸乳杆菌和干酪乳杆菌的计数；若产品中只含有嗜酸乳杆菌和干酪乳杆菌时，可先测其总数，再用LC—琼脂培养基选择计数干酪乳杆菌数，两者之差即为嗜酸乳杆菌数。     

Lactobacillus casei MYL01 modulates the proinflammatory state induced by ethanol in an in vitro model

Accumulating studies have suggested that probiotics have beneficial effects on liver injury but the underlying mechanism has remained unclear. Toll-like receptors (TLR) expressed on immune cells and hepatocytes recognize bacterial components that are translocated from the gut into the portal vein. To date, it has been demonstrated that ethanol alone, without microbial components, is able to activate TLR, leading to promotion of proinflammatory cytokine production. Because the enhanced signaling of TLR triggers persistent inflammation, we hypothesized that development of hepatocyte TLR tolerance to repetitive stimulation plays an important role in protecting the liver from hypergeneration of proinflammatory cytokines. In this study, we showed that Lactobacillus casei MYL01 modulated the proinflammatory state induced by ethanol and investigated in detail the mechanism underlying the observation that L. casei MYL01 gave rise to TLR tolerance toward ethanol stimulation. The effects of L. casei MYL01 in the attenuation of ethanol-induced liver damage were due to enhancement of IL-10 production, which limited the proinflammatory process. Furthermore, better defense of hepatocytes against ethanol challenge by treatment of L. casei MYL01 was attributed to previous induction of toll interacting protein (TOLLIP) and suppressor of cytokine signaling (SOCS)1 and SOCS3 expression via activation of TLR1, TLR2, TLR6, and TLR9, an action that cross-regulated ethanol–TLR4–nuclear factor κB signal transduction events. This finding might help establish an in vitro platform for selecting hepatoprotective probiotic strains in terms of ethanol-induced liver damage.     

Innovative production of multistrain synbiotic product using Thai-pigmented rice and rice bran oil

This study found that the isolated probiotics Lactobacillus paracasei KUKPS6201, L. acidophilus KUKPS6107, L. reuteri KUKPS6103, L. rhamnosus KUKPS6007, L. salivarius KUKPS6202, Bacillus coagulans KPSTF02 and Saccharomyces boulardii KUKPS6005 had high potential for probiotic properties. All strains had antibacterial activity and high antioxidant activity of 1.654 ± 0.017 mg Trolox mL⁻¹ probiotic extract. The selected strains could survive in a simulated gastrointestinal tract under anaerobic conditions and showed no haemolytic activity. Furthermore, the probiotic strains were strongly auto-aggregated and also showed co-aggregated ability with pathogenic bacteria. The probiotic microorganisms demonstrated high ability to adhere to Thai-pigmented rice grains. The results of analysis of these probiotics showed that Riceberry rice bran oil was an excellent prebiotic. A synbiotic product containing Thai-pigmented rice grains (cultivar Riceberry, Luem Pua and Black Jasmine) and rice bran oil was produced. After 8 weeks of storage, the viability of the probiotics in terms of multistrains was 7.36 ± 0.04 log CFU g^–1 (85.78% survival rate). Microbiological safety testing indicated that the amounts of contaminants were acceptable. This study provided the first scientific report on the feasibility of applying Thai-pigment rice, rice bran oil and mixed-culture probiotics as a novel functional synbiotic product.     

Viability of Lactobacillus acidophilus in rice bran‐enriched stirred yoghurt and the physicochemical and sensory characteristics of product during refrigerated storage

This study was aimed at investigating the fortification of probiotic yoghurt with rice bran to increase nutritional properties of the product. The different levels of rice bran (0.3%, 0.6%, 0.9% and 1.2%) were incorporated into milk. The yoghurt samples were produced after pasteurisation, addition of starter culture and 1% Lactobacillus acidophilus suspension (6 × 10⁸ CFU mL^?1) and incubation. During sample storage in refrigerator, the viability of L. acidophilus, viscosity and physicochemical and sensory properties of product were investigated. Rice bran significantly increased the viability of L. acidophilus (P < 0.05). In addition, all probiotic yoghurts incorporating rice bran indicated higher viscosity and acidity and lower pH and syneresis compared to plain yoghurts. Furthermore, increments in rice bran incorporation levels resulted in a reduction in consumers' sample preferences. In general, the addition of rice bran at a suitable level could increase L. acidophilus viability and improve quality attributes of yoghurt.     

Lactobacillus delbrueckii subsp. bulgaricus和Streptococcus thermophilus接种比例对Bifidobacterium lactis益生菌发酵乳品质的影响

将Lactobacillus delbrueckii subsp.bulgaricus ND02(LB-ND02)和Streptococcus thermophilus ND03(ST-ND03)按1∶1、1∶10、1∶100、1∶1000接种于脱脂乳中,同时接入益生菌Bifidobacterium lactis V9(B.lactis V9,接种量为2.0×107g-1),于42℃进行发酵。通过对发酵及贮藏过程中发酵乳指标的测定,评价LB-ND02和ST-ND03的接种比例对发酵乳品质的影响。结果表明,随着LB-ND02接种比例减小,凝乳时间显著延长,B.lactis V9活菌数显著提高。4℃贮藏28 d后,随LB-ND02接种比例减小,B.lactis V9存活率差异显著,后酸化也显著减弱。研究发现,LB-ND02和ST-ND03的接种比例,显著影响发酵乳的发酵时间、B.lactis V9活菌数、后酸化及黏度。     

Effect of different levels of fat and inulin on the microbial growth and metabolites in probiotic yogurt containing nonviable bacteria

Effects of different levels of fat and inulin on bacterial cell counts, degree of proteolysis and concentrations of organic acids in the yogurt containing inactivated cells of probiotic strains Bifidobacterium animalis and Lactobacillus acidophilus were investigated. Results showed that both L. acidophilus and B. animalis grew well in the yogurt samples reaching cell counts higher than 10⁶ CFU mL^?1 at the final pH of 4.5. Inulin at the concentration of 1% had no significant effects on the production of organic acids and cell counts of L. acidophilus, but promoted the growth of B. animalis with a reduction in the degree of proteolysis. Generally, different fat levels showed significant effects on the production of organic acids and nonsignificant effects on the cell counts of probiotic bacteria and degree of proteolysis. In case of lactic acid, the ratio of L‐ (+)to D‐ (?) isomer ranged from 50/50 to 80/20 in yogurt samples.     

Invited review: Potential antiobesity effect of fermented dairy products

The growing prevalence of obesity affects millions of people around the world and has gained increased attention over the years because it is associated with the development of other chronic degenerative diseases. Different organizations recommend lifestyle changes to treat obesity; nevertheless, other strategies in addition to lifestyle changes have recently been suggested. One of these strategies is the use of probiotics in fermented dairy products; however, a need exists to review the different studies available related to the potential antiobesity effect of these products. Because probiotic fermented dairy products that support weight management are not available in the market, there is a great opportunity for the development of functional dairy products with new lactic acid bacteria that may present this added health benefit. Thus, the purpose of this overview is to highlight the importance of probiotic fermented dairy products as potential antiobesogenic functional foods and present in vitro and in vivo studies required before this kind of product may be introduced to the market. Overall, most studies attributed the antiobesity effect of fermented dairy foods to the probiotic strains present; however, bioactive peptides released during milk fermentation may also be responsible for this effect.     

Microbiological,immunological, and histological changes in the gut of Salmonella Enteritidis-challenged rats fed goat cheese containing Lactobacillus rhamnosus EM1107

Cheeses are able to serve as suitable matrices for supplying probiotics to consumers, enabling appropriate conditions for bacteria to survive gastric transit and reach the gut, where they are assumed to promote beneficial processes. The present study aimed to evaluate the microbiological, immunological, and histological changes in the gut of Salmonella Enteritidis-challenged rats fed goat cheese supplemented with the probiotic strain Lactobacillus rhamnosus EM1107. Thirty male albino Wistar rats were randomly distributed into 5 experimental groups with 6 animals each: negative (NC) and positive (PtC) control groups, control goat cheese (CCh), goat cheese added with L. rhamnosus EM1107 (LrCh), and L. rhamnosus EM1107 only (EM1107). All animals, except NC group were challenged with Salmonella Enteritidis (10⁹ cfu in 1 mL of saline through oral gavage). Microbial composition was assessed with high-throughput 16S rRNA sequencing by means of Illumina MiSeq (Illumina, San Diego, CA). Nuclear factor kappa B (NF-κB) from the animal cecum tissue was determined by real-time PCR and interleukins (TNF-α, IL-1β, IL-10, and IFN-γ) by means of ELISA. Myeloperoxidase and malondialdehyde levels were determined biochemically. The administration of the L. rhamnosus EM1107 probiotic strain, either as a pure culture or added to a cheese matrix, was able to reduce Salmonella colonization in the intestinal lumen and lessen tissue damage compared with rats from PtC group. In addition, the use of cheese for the probiotic strain delivery (LrCh) was associated with a marked shift in the gut microbiota composition toward the increase of beneficial organisms such as Blautia and Lactobacillus and a reduction in NF-κB expression. These findings support our hypothesis that cheeses might be explored as functional matrices for the efficacious delivery of probiotic strains to consumers.