Multistrain Probiotics Alleviate Diarrhea by Modulating Microbiome-Derived Metabolites and Serotonin Pathway.

Diarrhea, a common gastrointestinal symptom in health problems, is highly associated with gut dysbiosis. The purpose of this study is to demonstrate the effect of multistrain probiotics (Sensi-Biome) on diarrhea from the perspective of the microbiome-neuron axis. Sensi-Biome (Lactiplantibacillus plantarum, Bifidobacterium animalis subsp. lactis, Lactobacillus acidophilus, Streptococcus thermophilus, Bifidobacterium bifidum, and Lactococcus lactis) was administered in a 4% acetic acid-induced diarrhea rat model at concentrations of 1 × 108 (G1), 1 × 109 (G2), and 1 × 1010 CFU/0.5 mL (G3). Diarrhea-related parameters, inflammation-related cytokines, and stool microbiota analysis by 16S rRNA were evaluated. A targeted and untargeted metabolomics approach was used to analyze the cecum samples using liquid chromatography and orbitrap mass spectrometry. The stool moisture content (p < 0.001), intestinal movement rate (p < 0.05), and pH (p < 0.05) were significantly recovered in G3. Serotonin levels were decreased in the multistrain probiotics groups. The inflammatory cytokines, serotonin, and tryptophan hydroxylase expression were improved in the Sensi-Biome groups. At the phylum level, Sensi-Biome showed the highest relative abundance of Firmicutes. Short-chain fatty acids including butyrate, iso-butyrate, propionate, and iso-valeric acid were significantly modified in the Sensi-Biome groups. Equol and oleamide were significantly improved in the multistrain probiotics groups. In conclusion, Sensi-Biome effectively controls diarrhea by modulating metabolites and the serotonin pathway.

Biosurfactant potential and antiviral activity of multistrain probiotics.

The COVID-19 caused by the SARS-CoV-2 has become a great threat to humans. However, there is no recommendation for an effective and safe drug to treat the disease. The strategy developed in this study is to utilize biosurfactant potential activity of Lactobacillus spp. and Rhodopseudomonas palustris probiotics to prevent the virus from entering human body. The outer membrane of the virus is comprising of phospholipid compounds. Biosurfactants, are known to have detergent-like properties (able to dissolve lipids) that are safe for in vivo use. Thus, the biosurfactant potential activity of the multistrain probiotics extract is expected to be able to disrupt the phospholipid membrane, resulting in the inactivity of the virus to infect human body. The biosurfactant potential activity of the probiotics extract was evaluated using oil spreading, drop collapse, and emulsification methods. The virus infectivity was evaluated on the SARS-CoV-2 of delta variant as a virus model. The results indicated that the probiotics extract has biosurfactant potential activity, able to inhibit virus growth up to 99.9 % within 48 h in the prevention platform, and up to 99.6 % within 48 h in the treatment platform. Therefore, the multistrain probiotics extract was identified to have potential as a promising antiviral agent.

New insights on strain-specific impacts of probiotics on insulin resistance: evidence from animal study.

BACKGROUND AND AIMS: сomparative animal study of effectiveness of intermittent administration of lyophilized single-, three- and alive multistrain probiotic in short courses on insulin resistance (IR) in rats with experimental obesity. METHODS: 70 rats were divided into 7 groups (n = 10 in each). Rats of group I were left intact. Newborn rats in groups II-VII were administered monosodium glutamate (MSG) (4 mg/g) by injection. Rats in group II (MSG-obesity group) were left untreated. The rats in groups III-V received lyophilized mono-probiotics B.animalis VKL, B.animalis VKB, L.casei IMVB-7280 respectively. The rats in group VI received all three of these probiotic strains mixed together. Group VII was treated with multi-probiotic "Symbiter", containing 14 different live probiotic strains (Lactobacillus, Bifidobacterium, Propionibacterium, Acetobacter genera). RESULTS: Treatment of newborn rats with MSG lead to the development of obesity in all MSG-obesity rats and up to 20-70% after probiotic administration. Additions to probiotic composition, with preference to alive strains (group VII), led to significantly lower rates of obesity, decrease in HOMA-IR (p < 0.001), proinflammatory cytokines levels - IL-1ß (p = 0.003), IL-12Bp40 (p < 0.001) and elevation of adiponectin (p = 0.003), TGF-ß (p = 0.010) in comparison with MSG-obesity group. Analysis of results in groups treated with single-strain probiotics (groups III-V) shows significant decrease in HOMA-IR, but changes were less pronounced as compared to mixture groups and did not achieve intact rats level. Other metabolic parameters were not affected significantly by single strains. CONCLUSION: Our findings provide major clues for how to design and use probiotics with more efficient compositions in obesity and IR management and may bring new insights into how host-microbe interactions contribute to such protective effects.

The Evolution of Human Probiotics: Challenges and Prospects.

In recent years, the intestinal microbiota has been found to greatly influence a number of biological processes important for human health and longevity. Microbial composition changes easily in response to external factors, such as an unbalanced diet, lack of physical activity, and smoking. Probiotics are a key factor in maintaining the optimal composition of the intestinal microbiota. However, a number of important questions related to probiotics, such as indication for prescription, comparative efficacy of monostrain and multistrain probiotics, methods of delivery, and shelf life, remain unresolved. The aim of this review is to highlight existing issues regarding probiotic production and their prescription. The review presents the most recent findings regarding advantages and efficacy of monostrain and multistrain probiotics, preservation of probiotic strains in capsules and microcapsules, production of probiotics in the form of biofilms for improved efficacy and survival, and results of clinical studies evaluating the benefits of probiotics against different pathologies. We believe that this work will be of interest to physicians and researchers alike and will promote the development of new probiotics and ensuing regimens aimed at the treatment of various diseases.

Variability in Probiotic Formulations Revealed by Proteomics and Physico-chemistry Approach in Relation to the Gut Permeability.

Variability in the efficacy, safety, and quality of probiotic formulations depends on many factors, including process conditions used by manufacturers. Developing reliable analytical tools is therefore essential to quickly monitor manufacturing differences in probiotic samples for their quality assessment. Here, multi-strain probiotics from two production sites and countries were investigated by proteomics and physico-chemistry approaches in relation to the protective effect on gut barrier. Proteomic analyses showed differences in protein abundances, identities, and origins of two series of VSL#3 samples from different sites. Even though both formulations were qualitatively similar in thermal and colloidal profiles, significant differences were quantitatively observed in terms of maximum decomposition temperature Tmax (p < 0.05) and phase transition temperature Tm (p < 0.01). Such variability in physical and biochemical features impacts on probiotic functionalities and translates into a differential modulation of gut permeability in mice. Physico-chemical scans provide coherent data with proteomics and represent a new tool for time and cost effective quality control of probiotic-based products.

The effect of supplementation of multistrain probiotic preparation in combination with vitamins and minerals to the basal diet on the growth performance, carcass traits, and physiological response of broilers.

AIM: The aim of the present study was to investigate the effect of supplementation of multistrain probiotic preparation in combination with vitamins and minerals to the broiler chicken diets on their growth performance, hematological parameters, and carcass traits. MATERIALS AND METHODS: Two hundred and eighty-eight Lohmann 1-day-old broiler chicks were randomly allocated to four groups, i.e., control (without additional supplementation) and three experimental treatments where basal diet was enriched by 0.1%, 0.5%, or 1% of multistrain probiotic preparation in combination with vitamins and minerals, respectively. Blood sampling was conducted on day 28, while the selected organs and eviscerated carcasses were collected on day 42. RESULTS: Dietary supplementation did not affect (p>0.05) the final body weight, feed intake, and feed conversion ratio of broilers. Supplementation by 0.1% and 0.5% of multistrain probiotic preparation in combination with vitamins and minerals reduced (p≤0.05) heart relative weight of broilers. Dietary supplementation tended (p=0.07) to increase the relative weight of ileum and pancreas of broilers. Supplemented birds had lower (p≤0.05) numbers of leukocytes and eosinophils compared to unsupplemented birds. There were tendencies that supplementation of multistrain probiotics in combination with vitamins and minerals resulted in lower (p=0.07) counts of lymphocytes and heterophils when compared with no supplementation. Supplementation by 0.5% of multistrain probiotics in combination with vitamins and minerals resulted in lower (p≤0.05) serum concentration of uric acid when compared with control. There was no significant effect of dietary supplementation on carcass traits, pH, and drip loss of broiler breast muscles. CONCLUSION: Dietary supplementation of commercial broiler feeds with 0.5% of multistrain probiotic preparation in combination with vitamins and minerals was potential to improve digestive functions and physiological status of broiler chickens.

Probiotics for experimental obesity prevention: focus on strain dependence and viability of composition.

INTRODUCTION: a comparative animal study of the efficacy of intermittent short-course administration of lyophilised single-, three-, and live multistrain probiotic on obesity. METHODS: We included 70 rats divided into seven groups (n = 10 in each). Rats of group I were intact. Newborn rats of groups II-VII were injected with monosodium glutamate (MSG) (4 mg/g). Rats of group II (MSG-obesity group) were untreated. The group III-V received lyophilised monoprobiotics B. animalis VKL, B. animalis VKB, and L. casei IMVB-7280, respectively. Group VI received the mix of these three probiotic strains. Group VII was treated with multiprobiotic "Symbiter", which contains 14 live probiotic strains (Lactobacillus, Bifi-dobacterium, Propionibacterium, Acetobacter genera). RESULTS: Neonatal treatment with MSG caused stunted growth, which is why, despite the lack of weight gain dynamics and absence of significant food consumption rate and body weight changes at day 120, we noted the development of obesity in all MSG-obesity rats and in up to 20-70% after probiotic administration. Supplementation of probiotic composition, with preference to live strains, led to a significantly lower prevalence of obesity, and reduction of VAT weight and serum lipid levels as compared to single-strain probiotic. In our comparative single-strain analysis a trend towards more pronounced hypolipidaemic effect and VAT weight reduction was observed for lyophilised L. casei IMVB-7280 as compared to B. animalis VKL and VKB strains. CONCLUSIONS: Multistrain formed mutualistic interactions in mixtures and therefore able to share with different metabolites, affect differ-ent receptors and produced various of biologically active compounds which synergistic overall effect greater than the sum of the single effects.

Effects of multistrain probiotics on growth performance, nutrient digestibility, blood profiles, faecal microbial shedding, faecal score and noxious gas emission in weaning pigs.

This experiment was conducted to investigate the efficacy of multistrain probiotics in weaning pigs. A total of 125 28-day-old weaning pigs [(Landrace × Yorkshire) × Duroc] with an initial average body weight (BW) of 7.26 ± 0.76 kg were randomly allotted into 5 treatments, 5 replicate pens/treatment with 5 pigs/pen for 42-day experiment. Dietary treatments were as follows: CON, basal diet; PC1, CON + 0.01% multistrain probiotics; PC2, CON + 0.03% multistrain probiotics; PC3, CON + 0.06% multistrain probiotics; PC4, CON + 0.1% multistrain probiotics. On day 14, pigs fed the PC4 diet had higher BW gain than pigs fed the CON diet. On day 42, pigs fed multistrain probiotics supplementation diets had higher BW gain than pigs fed the CON diet. From days 1 to 14, pigs fed the PC2, PC3 and PC4 diets had higher (p < 0.05) ADG than pigs fed the CON diet. From day 15 to 42, pigs fed the multistrain probiotics supplementation diets had higher (p < 0.05) average daily gain (ADG) and gain: feed ratio (G:F) than pigs fed the CON diet. In the overall period, pigs fed the multistrain probiotics supplementation diets had higher (p < 0.05) ADG and pigs fed the PC2 and PC4 diets had higher (p < 0.05) G:F than pigs fed the CON diet. On day 42, pigs fed the PC4 diet had higher (p < 0.05) apparent total tract digestibility (ATTD) of dry matter (DM), nitrogen (N) and gross energy (GE), faecal Lactobacillus counts and lower (p < 0.05) E. coli counts and NH3 emission than pigs fed the CON diet. Pigs fed the multistrain probiotics supplementation diets had lower (p < 0.05) H2 S and total mercaptans emissions than pigs fed the CON diet. Conclusions, dietary supplementation with 0.1% probiotics improved growth performance, nutrition digestibility and intestinal microflora balance and decreased faecal noxious gas emissions in weaning pigs.

Comparative experimental investigation on the efficacy of mono- and multiprobiotic strains in non-alcoholic fatty liver disease prevention.

BACKGROUND: To investigate the efficacy of different probiotic strains, their combinations and forms (alive or lyophilized) in nonalcoholic fatty liver disease (NAFLD) prevention. METHODS: In this study, 70 rats have been used divided into 7 groups of 10 animals in each: I - intact rats, II-VII - rats with monosodium glutamate (MSG)-induced NAFLD. Rats with NAFLD were untreated (group II, MSG-obesity group) and treated with probiotics (groups III-VII). In order to develop NAFLD, newborn rats of groups II-VII were injected with a solution of monosodium glutamate (MSG) (4 mg/g) subcutaneously (s.c.) at 2nd,4th, 6th, 8th,10th postnatal day. The groups III-V received lyophilized monoprobiotics B. animalis VKL, B. animalis VKB, L.casei IMVB-7280, respectively. The group VI received 2.5 ml/kg of an aqueous solution of a mixture of the three probiotic strains (2:1:1 Lactobacillus casei IMVB-7280, Bifidobacterium animalis VKL, Bifidobacterium animalis VKB) at a dose of 50 mg/kg (5 × 10(9) CFU/kg) (g) (intragastrically). The group VII was treated with multiprobiotic "Symbiter" containing biomass of 14 alive probiotic strains (Lactobacillus + Lactococcus (6 × 10(10) CFU/g), Bifidobacterium (1 × 10(10)/g), Propionibacterium (3 × 10(10)/g), Acetobacter (1 × 10(6)/g)) at a dose of 140 mg/kg (1.4 × 10(10) CFU/kg). The treatment with probiotics was started at the age of 1 month. There were 3 courses of treatment, each included 2-week administration and 2-week break. All parameters were measured in 4-month aged rats. RESULTS: Introduction of MSG during the neonatal period leads to the NAFLD development in the 4-months old rats. For steatosis degree there was no significant difference between MSG-obesity group and lyophilized monocomponent probiotics groups (III-V). The highest manifestation of steatosis was observed for B. animalis VKL group (2.0 ± 0.25) as compared to B. animalis VKB (1.70 ± 0.21) and L. casei IMVB-7280 (1.80 ± 0.20). The steatosis score changes between all monoprobiotics groups (III-V) were insignificant. Administration from birth of both alive (VII) and lyophilized (VI) probiotic mixture lead to a significant decrease by 69.5 % (p < 0.001) and 43.5 % (p < 0.025) of steatosis score respectively as compared to the MSG-obesity group (2.3 ± 0.21 %). For both alive and lyophilized probiotic mixtures, reduction of lobular inflammation was observed. These histological data were confirmed by the significant decrease of total lipids and triglycerides content in the liver approximately by 22-25 % in groups treated with probiotic mixtures (VI, VII) compared to the MSG-obesity group. CONCLUSION: We established failure of NAFLD prevention with lyophilized monoprobiotic strains and the efficacy of probiotic mixture with the preference of alive probiotic strains.