Strains Of Probiotic Bacteria Research Articles

Effect of the Bacillus subtilis BS20 on physiological and immune parameters in mutant mice Muc2–/–

Introduction. Inflammatory bowel disease (IBD) is a common pathology that cannot be completely cured and requires lifelong therapy. One of the promising and sparing therapeutic strategies is the use of probiotics. Unlike Lactobacillus and Bifidobacterium, which represent the basis of most classical probiotics, Bacillus spp. are spore-forming bacteria that contributes to their resistance to the acidic environment of the stomach. It has been shown that the bacteria Bacillus spp. is an effective therapeutic drug for the relief of intestinal diseases with signs of inflammation and bacterial origin. Since the main factors causing IBD are dysbiosis and inflammation, and strains of Bacillus spp. as probiotics can be potential modulators of immunity and influence the intestinal microbiome, it is necessary to search for new strains of probiotic bacteria, including representatives of Bacillus spp., which have an effect on immunity and physiological indicators in experimental models of IBD. The aim of the scientific research — to study the effect of Bacillus subtilis BS20 spores on the physiological and immune parameters of the mutant mouse line Muc2–/–. Research objectives: Investigate survival of Muc2–/– mice, weight, immune parameters (cytokines and immune cells) and amino acid composition of muscles. Materials and methods. Muc2–/– females were fed spores of Bacillus subtilis BS20 in the amount of 109 CFU for 2 months. Analysis of the amino acid composition of the femoral muscle was performed by capillary electrophoresis. The concentration of cytokines in the supernatant was measured using the Magnetic Luminex assay kit. Determination of the number of lymphocytes was performed by flow cytometry. Results. The addition of Bacillus subtilis BS20 to the diet of mice reduced mortality and body weight loss in Muc2–/– females. We found a decrease in interleukin-6 and an increase in interleukin-17 in the descending colon and an increase in B and T helper lymphocytes in the spleen. Conclusion. B. subtilis BS20 improves the general condition of Muc2–/– mutant mice, has an anti-inflammatory and immunostimulating effect, reducing the level of the cytokine interleukin-6 and increasing the percentage of B cells and T helper cells in the spleen.

Interrelations Between Probiotics, Gut Microbiota, Intestinal Barrier, and Immune Response; Focusing on Diarrhea in Dairy Calves1

The interplay between gut microbiota and host health has attracted significant interest in the animal science community. Maintaining gut microbiota homeostasis by supplementing probiotics to treat clinical conditions like calf diarrhea is an emerging area of research nowadays because of increased concerns regarding antimicrobial resistance (AMR) and drug residues in animal products. Probiotics reduce the incidence of calf diarrhea by increasing the gut microbiota diversity and richness with more commensal bacteria such as Lactobacillus and Bifidobacterium that produce antimicrobial compounds, as well as modulating the immune response by increasing cytokines, Interleukin-2 (IL-2), IL-4, IL-6, IL-10, and reducing tumor necrosis factor-α (TNF-α), by increasing production of antibodies, especially immunoglobulin E (IgE), also IgG, differentiating naïve Th lymphocytes (Tho) into Th1, hence stimulate innate immunity and prime the adaptive immune response. Specific probiotic strains of bacteria and yeast (Saccharomyces cerevisiae) derived probiotics maintain the integrity of the intestinal barrier. In this review, data is being organized to address the role of probiotics in treating calf diarrhea by modulating gut microbiota and stimulating an immune response against notorious pathogens, to present animal and veterinary scientists and nutritionists with a new concept to treat infectious diseases from the perspective of the gut microbiota, increasing animal health, performance, and welfare. In conclusion, health status and gut microbiome are strongly interlinked. Research data indicated a significant reduction in the incidence of diarrhea after probiotic administration. If interrelations between probiotics and existing gut microbiota are explored more quantitatively, novel antibiotic substitutes can emerge in the future.

Abstract 5237: Engineered probiotics direct the antigen specificity of CAR-T cells in situ

Abstract Antigen-targeting therapies such as chimeric antigen receptor (CAR)-T cells have achieved unparalleled success in the treatment of hematological malignancies. However, broad success has remained bottlenecked by the lack of targets that are specifically, and uniformly, expressed on heterogenous solid tumors. In contrast, certain strains of bacteria are gaining recognition as a new class of antigen-agnostic cell therapies due to their selective growth within the immuno-suppressive niche of the solid tumor microenvironment (TME). Bridging these approaches, we developed a platform of probiotic-guided CAR-T cells (ProCARs) - in which tumor-colonizing probiotics release synthetic targets that efficiently tag tumor tissue for CAR-mediated lysis. Here, we engineered a well-characterized strain of probiotic bacteria, E. coli Nissle 1917, to sustain the intratumoral production of genetically encoded CAR targets and T cell attracting chemokines in multiple xenograft and syngeneic models of human and murine tumors. We design orthogonal CAR targets (Tags) as modified dimers of GFP that broadly anchor to collagens, fibronectins, and heparin sulfates found in high abundance within the solid TME to achieve antigen-agnostic tumor targeting. We demonstrate that Tags robustly coat the surface of cancer cell lines through interaction with cell surface matrix proteins, thus leading to the activation of GFP CAR-T cells and Tag-dependent killing across a panel of genetically distinct target cells. We additionally show that injected probiotics selectively grow within the tumor core and maintain the intratumoral production of Tags and human chemokines - leading to therapeutic efficacy across multiple subcutaneous and orthotopic tumor models. Moreover, we demonstrate a systemic antitumor benefit in immune-competent hosts, whereby unilateral treatment of primary tumors leads to a significant reduction in the growth rate of distal, untreated tumors. Finally, we show that intratumoral bacteria provide natural TLR agonists that trigger substantial activation of human and murine ProCAR-T cells. Our findings highlight the potential of the ProCAR platform to address the critical roadblock of identifying suitable CAR targets by providing an antigen in situ that is orthogonal to both healthy tissue and tumor genetics. Altogether, the use of a bacterial delivery platform in the ProCAR system offers a partner organism that naturally enhances CAR-T cell effector function and broadens the scope of CAR-T cell therapy to include previously difficult to target tumors. Citation Format: Rosa L. Vincent, Fangda Li, Ana Vardoshvili, Candice Gurbatri, Nicholas Arpaia, Tal Danino. Engineered probiotics direct the antigen specificity of CAR-T cells in situ [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5237.

Evaluation of probiotic properties of a Brevibacillus laterosporus strain.

Brevibacillus laterosporus is a strain of probiotic bacteria that has been widely used in pest control, cash crop, and other production areas. However, few studies have been conducted on its use as a feed additive in animals. Therefore, the probiotic potential of B. laterosporus PBC01 was evaluated by characterizing hydrophobicity, auto-aggregation activity, bile salt and simulated gastrointestinal fluid tolerance, bienzymatic, and antibacterial activity. Antibiotic susceptibility, hemolysis assays, and supplemental feeding of mice were also performed to evaluate safety features. Our results showed that B. laterosporus PBC01 had moderate hydrophobicity, high auto-agglutination ability. Meanwhile, B. laterosporus PBC01 had good tolerance to bile salt and simulated gastrointestinal fluid. It had the ability to secrete protease, cellulase, and to inhibit various pathogens. In addition, B. laterosporus PBC01 was sensitive to many antibiotics, and did not produce hemolysin. In the safety assessment of mice, it did not cause any deaths, nor did it affect the cell components of blood, antioxidant capacity, and reproductive health. The study indicated the great probiotic characteristics and safety of B. laterosporus PBC01. This may provide a theoretical basis for the clinical application and development of probiotic-based feed additives.

APPLICATION OF WHEY PROTEIN HYDROLYSATES AS A FERMENTATION MEDIUM WITH STRAINS OF LACTIC ACID BACTERIA

The aim of the present study is to obtain enzymatic hydrolysates from whey protein and evaluate their potential application as a substrate for fermentation with lactic acid bacteria. For hydrolysis of whey proteins, papain is used in 3 concentrations - 0.1, 0.5 and 1.0 mg mL-1. The degree of hydrolysis (DH) was determined on the obtained whey protein hydrolysates (WPHs), the highest percentage being achieved with hydrolysate WPH3 using the papain enzyme at a concentration of 1 mg mL-1. Fermentation of the whey protein hydrolysates was carried out with the participation of the probiotic strains Lactiplantibacillus plantarum subsp. plantarum NBIMCC 3447 and Lactobacillus gasseri NBIMCC 2450. The survival of two strains of probiotic lactic acid bacteria was investigated. In the samples obtained from hydrolyzed whey medium with 0.5 mg mL-1papain, the highest values of viable probiotic bacterial cells were recorded - for Lactiplantibacillus plantarum NBIMCC 3447 2.5 х 109 CFU mL-1, and for Lactobacillus gasseri NBIMCC 2450 2.1 х 109 CFU mL-1, respectively. Antioxidant activity is reported on the fermented products. The WPH2 variant has the highest antioxidant potential - 9.14 mg TE/100 mL.

Ligilactobacillus Salivarius improve body growth and anti-oxidation capacity of broiler chickens via regulation of the microbiota-gut-brain axis

Certain strains of probiotic bacteria can secret functional substances namely digestive enzymes and functional peptides to regulate physiological conditions such as digestion and anti-oxidation, which are often incorporated in industrial broiler chick production. However, few studies have detailed the action mechanisms and effects of these bacteria on regulating growth and anti-oxidation levels in broiler chickens. Ligilactobacillus salivarius is a strain of probiotic bacteria used as dietary supplement. In the present study, Ligilactobacillus salivarius was evaluated for its secreted digestive enzymes in vitro. To detailed evaluate the action mechanisms and effects of gastrointestinal tract (GIT) microbiota on alleviating anti-oxidation levels of broiler chickens through the gut-brain axis. Ligilactobacillus salivarius was cultured and supplemented in the food of broilers to evaluate the probiotic effect on growth and anti-oxidation by modulation of gut microbial composition and its functional metabolites using metagenomic and metabolomic assays. Biochemical results showed that Ligilactobacillus salivarius secreted digestive enzymes: protease, lipase, and amylase. Broiler chickens with Ligilactobacillus salivarius supplemented for 42 days, showed increased body weights, a reduced oxidative status, decreased malondialdehyde levels, and improved activities rates of total superoxide dismutase, glutathione peroxidase IIand IV improved. The microbial composition of caecum was more abundant than those broiler without probiotics supplementation, owing 400 of total number (489) of bacterial operational taxonomic units (OTU). The genera of Lactobacillus, Megamonas, Ruminoccoccaceae, Ruminococcus, Alistipes and Helicobacter shared the dominant proportion of Candidatus _Arthromitus compared with the control chickens. These functional bacteria genera assisted in the transportation and digestion of amino acids, carbohydrates, and ions, synthesis of cellular membranes, and anti-oxidation. Uncultured_organism_g_ Anaerosporobacter, Lactobacillus salivarius, uncultured_bacterium_g_ Ruminococcaceae_UCG-014, uncultured_bacterium_g_ Peptococcus were strongly and positively correlated with body growth performance and anti-oxidation. A metabonomic assay suggested that the secreted of gamma-aminobutyric acid and monobactam was metabolized according to the Kyoto Encyclopedia of Genes and Genomes analysis. In conclusion, Ligilactobacillus salivarius optimized microbial composition of the caecum and secreted functional peptides through gut-brain axis to improve the body growth and antioxidation of broiler chicken.

Isolation, identification, and molecular characterization of probiotic bacteria from locally selected Ethiopian free range chickens gastrointestinal tract

The poultry industry is facing continuous challenges with regard to increased feed costs and loss due to infectious disease. To overcome this challenge, several antibiotics have been used along with chicken feeds to promote growth. Nevertheless, the use of antibiotics as growth promoter has been banned in many countries, due to the concerns associated with potential risks of emerging and horizontal transfer of multidrug resistant genes to bacteria in animal tissues. The objective of this study was to identify and characterize potential probiotic bacteria strains from the gastrointestinal tract of free-range locally selected chickens. The bacterial isolates were screened, purified and characterized based on morphological, biochemical and molecular characteristics from 12 well adopted free-range healthy young chickens. Low pH and bile salt tolerance, antagonistic activity, antibiotic activity, hemolysis activity, adhesion to the chicken intestine and carbohydrate fermentation tests was conducted to identify potential probiotic bacteria. Twelve bacterial isolates were screened based on their ability for their tolerance to low pH and bile salt. The isolates were identified by using 16S rRNA gene partial sequencing method. All screened isolates showed great survival percentage at low pH, i.e. (89.2±0.75 to 97.1±0.64) survived at 3hrs and (83.6±0.75 to 95.2±0.63) at 6hrs challenge at pH2. Isolate GCM112 was the least tolerant strain in 6.0% salt concentration at 12 and 24hrs exposure time (82.1±1.28 and 79.9±1.96%) respectively. The result revealed no strain tests in this study exhibited α- and β-hemolytic activity when cultured in sheep blood agar. Most isolated strains showed best growth at 37°C temperature and up to 4% NaCl concentration. Based on the reported result from in vitro data, GCH212 and GCM412 isolates were recognized as best potential probiotic bacteria for chicken against pathogens but further studies are needed on in vivo assessment on the health benefits in the real life situation.

Natural resistance and immunological response of young Volyn meat breed to the action of probiotics

To investigate the response of natural resistance and immunological reactions of newborn young animals of the Volyn beef breed to the use of probiotic preparations. To conduct experimental research, three groups were formed from young Volyn meat breeds: the control group was fed only on mother's milk, the first group received a probiotic preparation with Bacillus Subtilis strain in addition to cow's milk, the second experimental group –– Lactobacillus spp. The activity of natural resistance and the body's immune response were studied on the 5th, 10th, and 30th day of the experiment. The level of nonspecific natural resistance was determined by indicators of bactericidal, lysozyme, phagocytic, and complementary activity of blood serum, and the immunological response was determined by the concentration of immunoglobulins of classes G, M, and A in the blood. It was established that starting from the 5th day of life, a difference in the nonspecific resistance of control and experimental animals was noted groups In separate age periods, probiotic supplements had a different degree of influence on the natural resistance of young animals. The greatest difference in the direction of growth in such indicators of non-specific resistance as bactericidal and complementary activity of blood serum was noted under the influence of the probiotic supplement Bacillus Subtilis (I experimental group), and indicators of lysozyme and phagocytic activity of blood serum were greater under the action of Lactobacillus spp. (II experimental group). The difference between indicators of bactericidal activity of blood under the action of Lactobacillus spp. in comparison with the control was at the age of 30 days (16.85 %, Р<0.01), and under the action of Bacillus Subtilis –– at the age of 60 days (25.49 %, Р<0.001). The level of phagocytic and complementary blood activity was the highest at the age of 60 days in the I experimental group –– by 17.17 % (Р<0.001) and 32.57 % (Р<0.001), in the II experimental group –– by 23.20 % (Р <0.001) and 36.34 % (Р<0.001), respectively. Regarding the concentration of immunoglobulins, the largest changes in their indicators were noted in the group that received Lactobacillus spp. Addition of probiotic strains of bacteria to the diet of young animals has a positive effect on the natural resistance and immune response of the body, which in the future will contribute to reducing the level of morbidity in young animals. Key words: calves, probiotics, bacterial strains, microorganisms, immunoglobulins, non-specific resistance of the organism, T-lymphocytes, phagocytic activity, bactericidal activity of blood serum.

Isolation and Identification of Dextran-Producing Weissella cibaria from Pickled Sauerkraut

The presence of certain bacteria in food products are particularly desirable. Probiotic strains of lactic acid bacteria (LAB), which have a number of favorable characteristics, including antimicrobial properties that support the development of beneficial microflora, are among them. In addition, because of their biochemical activity, they have an impact on the dietary, nutritional, and organoleptic qualities of food products. Before being released into the market, each strain must undergo independent testing to ascertain its probiotic qualities and potency. The purpose of this study was to separate and identify the LAB strain (Weissella cibaria bacteria) at the molecular level from a sample of pickled Sauerkraut that might be employed in the dairy sector and maybe is regarded as a probiotic in the future. Directly from the can, the shredded picked sauerkraut was brined in 2.25% salt (NaCl) for 48 hours before being inoculated aseptically into deMan, Rogosa, and Sharpe agar (MRS) and incubated for two days at 37 °C. Gram’s reaction, catalase production, and sugar fermentation tests employing glucose, sucrose, maltose, lactose, and fructose were among the traditional assays performed on isolation. The sequence was acquired and checked against those in the GenBank database using the BLAST tool at the NCBI-GenBank database after the isolate was extracted using the AccuPrep Genomic DNA Extraction kit, amplified, and agarose gel electrophoresis. The isolated LAB strains were validated using SEM and a genetic PCR assay, and the matching rate of the two isolated strains was 99%. The obtained strains showed their ability to produce dextran by observing the viscous growth characteristic when they were grown in MRS medium supplemented with 15% sucrose. To identify the bacterium, many culture and biochemical tests were carried out. The overall study concluded that the strains of Weissella cibaria bacteria can be easily isolated from pickled Sauerkraut using the developed procedure with the possibility of using the dextran produced by the bacteria as a prebiotic in the food sector.

The role of probiotics in the treatment of depressive disorders. A critical review

Depressive disorders are a widespread problem in modern medicine. According to current data from the World Health Organisation, an estimated 280 million people worldwide suffer from depression. In recent years, there have been reports of a correlation between the composition of the gut microbiota and the development of depressive disorders and attempts to modify it through the use of psychobiotics.
 The literature from the PubMed database published between 2018-2023 has been explored. 596 articles were selected based on the keywords "probiotics" and "depression". Six randomized clinical trials were finally included in the analysis.
 As defined elsewhere, psychobiotics are probiotic bacteria which supplemented in adequate amounts, interact with the gut-brain axis and show beneficial effects on patients' mental health. Results from recent RCTs suggest that daily probiotic supplementation significantly reduces the severity of depression compared to placebo (p<0.05). Additionally, this effect may be enhanced by the combined use of a probiotic with a prebiotic. Furthermore, some researchers indicate that probiotics may lead to significant improvements in cognitive function in patients suffering from depressive disorders.
 In conclusion, intestinal dysbiosis may be an important factor leading to the development of mental illness. Results of recent studies suggest that specific strains of probiotic bacteria may offer therapeutic benefits in the treatment of the aforementioned disorders. However, further clinical studies are needed to objectively confirm the relationship between gut microbiota composition and the development of depression.

A review and comparative perspective on health benefits of probiotic and fermented foods

SummaryFermented foods such as yogurt, kefir and sauerkraut have been part of the human diet throughout history and have gained attention in recent years due to their immense health and nutritional benefits. As a result, fermented foods are considered biofuel for the human microbiome which helps to boost the immune system. Fermented foods are those foods and beverages that are produced by employing specific microbial‐based fermentation aids such as yeasts and bacteria, particularly lactic acid bacteria (LAB). Through controlled enzymatic reactions, these microbial cultures transform food components as substrates into value‐added products promoting various healthy fermentative activities. These microbes in fermented foods also produce compounds that can inhibit food spoilage and pathogenic microorganisms, thereby extending the product shelf of fermented products. Probiotics are live microbiota with beneficial health properties, prevent gastrointestinal diseases and modulate the human microbiome. Thus, foods that are fermented by certain strains of probiotic bacteria that exhibit evidence of health benefits are referred to as probiotic fermented foods. This review describes fermented and functional foods, probiotics and their relationship to human health. In addition, we offer our perspective on the distinct differences between probiotic and fermented foods to promote awareness for consumers and key stakeholders regarding these highly functional and nutritionally fermented food products.

The Enzymatic Hydrolysis of Human Milk Oligosaccharides and Prebiotic Sugars from LAB Isolated from Breast Milk.

Breastfeeding is essential in the first months of a newborn's life. Breast milk is a source of crucial macronutrients, prebiotic oligosaccharides, and potential probiotic strains of bacteria. Oligosaccharides from breast milk (HMOs) are a significant part of the composition of breast milk and represent a complex of digestible sugars. This study aims to elucidate the enzymatic hydrolysis of these oligosaccharides and other prebiotics by the bacteria present in breast milk. We used modified methods to isolate oligosaccharides (HMOs) from human milk. Using unique techniques, we isolated and identified different bacteria from breast milk, mainly Lactobacillus fermentum. Using enzymatic analyses, we established the participation of α-fucosidase, α-glucosidase, β-galactosidase, and β-glucosidase from breast milk bacteria in the hydrolysis of prebiotic sugars. We also optimized the scheme for isolating oligosaccharides from breast milk by putting the lyophilized product into different food media. We found that the oligosaccharides from breast milk (HMOs) are a potent inducer for the secretion of the studied bacterial enzymes. Also, we found that all the lactobacilli strains we studied in detail could digest mucin-linked glycans. The degradation of these sugars is perhaps a built-in defense mechanism in cases where other sugars are lacking in the environment. We also determined fucosidase activity in some of the isolated strains. We recorded the highest values (2.5 U/mg in L. fermentum ss8) when the medium's oligosaccharides isolated from breast milk were present. Lactobacilli and Bifidobacteria supplied with breast milk are the first colonizers in most cases in the gastrointestinal tract of the newborn. The presence and study of different genes for synthesizing other enzyme systems and transporters of various sugars in this type of bacteria are essential.

Probiotic Fermented Goat's and Sheep's Milk: Effect of Type and Dose of Collagen on Survival of Four Strains of Probiotic Bacteria during Simulated In Vitro Digestion Conditions.

Microbial tolerance of digestive stresses depends not only on the bacterial strain but also on the structure and physicochemical properties of the supply chain and the foods that contain it. In the present study, we aimed to evaluate the effects of the type of milk (ovine, caprine) and the type and dose of collagen on the viability of four probiotic strains, Lacticaseibacillus paracasei L-26, Lacticaseibacillus casei 431, Lactobacillus acidophilus LA-5, and Lacticaseibacillus rhamnosus Lr-32, during in vitro gastrointestinal digestion. The highest survival rate under simulated in vitro digestion conditions compared to the number of cells before digestion was found in two strains, L. casei and L. paracasei, where survival rates were greater than 50% in each batch. The survival rate of the L. rhamnosus strain ranged from 41.05% to 64.23%. In caprine milk fermented by L. acidophilus, a higher survival rate was found in milk with 1.5% hydrolysate than the control, by about 6%. Survival of the L. rhamnosus strain was favorably affected by the 3% addition of bovine collagen in caprine milk, which increased survival by about 14% compared to the control sample. Adding 3% of hydrolysate to sheep's and goat's milk enhanced the survival of the L. rhamnosus strain by 3% and 19%, respectively. This study reports that fermented caprine and ovine milk may be suitable matrices for the probiotic supply of commercial dairy starter cultures and promote gut homeostasis.

Effect of oat bran addition on the survival of selected probiotic strains in Turkish fermented sausage during cold storage

The aim of this study was to determine the probiotic survivability of selected probiotic in the oat-bran added sausages. In this study, traditional fermented Turkish sausage was produced by adding oat bran at levels of 0.0%, 1.0%, 2.5%, and 5.0% of raw meat, as well as inoculating it with probiotic bacteria strains Lactobacillus acidophilus DSM 20079, Lacticaseibacillus casei 431®, and Lactobacillus acidophilus NCFM. The effects of these additions on the sausage were investigated by creating 16 different groups of samples (4 groups for each level of oat bran and control group (no inoculation) + 3 groups inoculated with different probiotic strains). After fermentation, the inoculated sausages were stored under highly controlled conditions for 30 days. Sausage samples underwent physicochemical and microbiological analyses at different storage stages (days 0, 15, and 30). It was found that the survival of probiotic bacteria was at an acceptable level (8–9 log10 CFU/g). The addition of oat bran and the inoculation of probiotic bacteria increased the thiobarbituric acid (TBARS) number of the sausage. It has been found that a 5% oat bran content contributes to the survival of L. acidophilus DSM 20079 and reduces the number of yeasts and moulds. L. acidophilus DSM 20079 also had the longest life cycle when compared to the other probiotics. Despite probiotic bacteria's potential for increasing oxidation, certain study groups showed lower TBARS values. To further improve their functional properties, it may be beneficial to consider using different starter cultures or a combination of starter cultures.

Bifidobacterium bifidum prevents the IL-1B induced increase in intestinal permeability by a novel mechanism: TLR-2 dependent activation of PPAR-gamma and inhibition of NF-kB signaling pathway

Background: Bifidobacterium bifidum (BB) is one of the most common probiotic bacteria strains that have been shown to have protective effects against intestinal inflammation, primarily by enhancing the intestinal epithelial tight junction (TJ) barrier function. IL-1β contributes to the development of intestinal inflammation in inflammatory bowel disease (IBD) in part by causing an increase in intestinal permeability. The IL-1β induced increase in intestinal epithelial TJ permeability is mediated by an NF-κB-dependent activation of myosin light chain kinase (MLCK) gene and disruption of the intestinal TJ barrier. The peroxisome proliferator-activated receptor gamma (PPAR-γ) is an important nuclear receptor in enterocytes which is known to have anti-inflammatory activity by interfering with NF-κB activation. However, the role of BB/PPAR-γ (and the possible mechanism involved) in protecting against IL-1β increase in intestinal permeability remain unclear. Aims: The major purpose of this study was to delineate the protective effect of BB against the IL-1β induced increase in intestinal TJ permeability and the intracellular mechanism involved. Methods: Filter-grown Caco-2 monolayers ( in vitro) and recycling intestinal perfusion of live mice ( in vivo) were used to assess intestinal permeability by using a paracellular marker (dextran-10kDa). Results: IL-1β caused a rapid activation of NF-κB and NF-κB and MLCK-dependent increase in intestinal epithelial TJ permeability in vitro and in vivo. A specific strain of BB (1x108 CFU/ml) referred to as BB1 strain inhibited the IL-1β increase in intestinal TJ permeability while strain BB4 had no effect. Other BB strains examined (5 total strains including BB4) did not inhibit the IL-1β increase in intestinal TJ permeability, suggesting that the BB enhancement of Caco-2 TJ barrier function was strain-specific. BB1, not BB4 enhancement of the TJ barrier was associated with an increase in TLR-2 expression, and PPAR-γ activation; BB1 also inhibited NF-kB activation and MLCK activity in Caco-2 monolayers and in mouse enterocyte. The inhibitory effect of BB on 1L-1β induced increase intestinal permeability, NF-κB activation, MLCK expression and activity was blocked by siRNA-induced knockdown of TLR-2 or PPAR-γ in Caco-2 monolayers. We also generated Villin-cre intestinal epithelial specific PPAR-γ knockout mice to study the involvement of enterocytes TLR-2/PPAR-γ on BB1 effect. BB1 did not inhibit the IL-1β-induced activation of NF-κB, increase in MLCK expression and increase in mouse intestinal permeability in TLR-2 or PPAR-γ intestinal deficient mice. Conclusion: These studies provide a novel insight into the BB the protective mechanism against the IL-1β-induced increase in intestinal TJ permeability in -vitro and in-vivo. Our data show that BB protects against the IL-1β induced increase in intestinal TJ permeability by a novel mechanism involving TLR-2/PPAR-g mediated inhibition of NF-kB and MLCK gene activatiation. NIH funding This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Prevalence of Aflatoxins in Camel Milk from the Arabian Peninsula and North Africa: A Reduction Approach Using Probiotic Strains.

Camel milk is known as a source of nutritional and health supplements. It is known to be rich in peptides and functional proteins. One main issue facing it is related to its contamination, mainly with aflatoxins. The present study aimed to evaluate camel milk samples from different regions while trying to reduce its toxicity using safe approaches based on probiotic bacteria. Collected samples of camel milk were sourced from two main regions: the Arabic peninsula and North Africa. Samples were tested for their contents of aflatoxins (B1 and M1) using two techniques to ensure desired contamination levels. Additionally, feed materials used in camel foods were evaluated. Applied techniques were also tested for their validation. The antioxidant activity of camel milk samples was determined through total phenolic content and antioxidant activity assays. Two strains of probiotic bacteria (Lactobacillus acidophilus NRC06 and Lactobacillus plantarum NRC21) were investigated for their activity against toxigenic fungi. The result revealed high contamination of aflatoxin M1 for all samples investigated. Furthermore, cross-contamination with aflatoxin B1 was recorded. Investigated bacteria were recorded according to their significant inhibition zones against fungal growth (11 to 40 mm). The antagonistic impacts were between 40% and 70% against toxigenic fungi. Anti-aflatoxigenic properties of bacterial strains in liquid media were recorded according to mycelia inhibition levels between 41 to 52.83% against Aspergillus parasiticus ITEM11 with an ability to reduce aflatoxin production between 84.39% ± 2.59 and 90.4% ± 1.32 from media. Bacteria removed aflatoxins from the spiked camel milk in cases involving individual toxin contamination.

75 Effect of probiotic supplementation on intestinal permeability in subjects with overweight and obesity: A systematic review of randomized controlled trials

OBJECTIVES/GOALS: Obesity is associated with gut dysbiosis, inflammation, and increased intestinal permeability. Probiotic consumption may reverse these outcomes. The goal of this study is to evaluate the evidence linking probiotic consumption to changes in intestinal permeability in subjects with overweight or obesity. METHODS/STUDY POPULATION: Articles were searched in Pubmed, Web of Science, and CAB Direct through February 2022 using search terms: intestinal permeability, overweight or obesity, and probiotic supplementation. 694 articles were exported, and 289 duplicates were identified. Titles and abstract were screened in the 405 remaining references by two investigators to determine eligibility. Eligible studies had data extracted on study participant characteristics, probiotic strain used, probiotic dosage, length of intervention, and intestinal barrier outcomes. Results were summarized in tabular form based on intestinal permeability response to probiotics. Quality of the studies was assessed based on Cochrane–Risk of Bias’ tool (RoB2). RESULTS/ANTICIPATED RESULTS: Thirteen eligible studies were identified. Probiotic genera included Akkermansia, Bifidobacterium, Lactobacillus, Streptococcus, Lactococcus, and Bacillus. Single strain probiotics were used in 3 studies, while the other 10 used multi-strain formulas. Dosage and length of probiotic supplementation ranged from 2.4 x 10^7 to 5 x 10^10 CFU/person/day and 3 to 26 weeks, respectively. The most widely used gut permeability outcomes were serum lipopolysaccharide (LPS) (n=10) and mixed sugar solution consumption with urine analysis (n=6). Five of the 10 studies reported decreases in serum LPS following probiotic consumption, while the other 5 showed no effect. Urinary lactulose/mannitol ratio decreased in 1 of the 4 studies, and urinary lactulose percent decreased in 2 studies. DISCUSSION/SIGNIFICANCE: Probiotic supplementation may be remediating an obesity-induced increase in intestinal permeability as evidenced from the effect on serum LPS and mixed sugar solution assays. However, additional studies are needed to further clarify which strain of probiotic bacteria is most effective and the optimal intervention length in subjects with obesity.

Preparation and Evaluation of a Dosage Form for Individualized Administration of Lyophilized Probiotics

Probiotics have been used in human and veterinary medicine to increase resistance to pathogens and provide protection against external impacts for many years. Pathogens are often transmitted to humans through animal product consumption. Therefore, it is assumed that probiotics protecting animals may also protect the humans who consume them. Many tested strains of probiotic bacteria can be used for individualized therapy. The recently isolated Lactobacillus plantarum R2 Biocenol™ has proven to be preferential in aquaculture, and potential benefits in humans are expected. A simple oral dosage form should be developed to test this hypothesis by a suitable preparation method, i.e., lyophilization, allowing the bacteria to survive longer. Lyophilizates were formed from silicates (Neusilin® NS2N; US2), cellulose derivates (Avicel® PH-101), and saccharides (inulin; saccharose; modified starch® 1500). They were evaluated for their physicochemical properties (pH leachate, moisture content, water absorption, wetting time, DSC tests, densities, and flow properties); their bacterial viability was determined in conditions including relevant studies over 6 months at 4 °C and scanned under an electron microscope. Lyophilizate composed of Neusilin® NS2N and saccharose appeared to be the most advantageous in terms of viability without any significant decrease. Its physicochemical properties are also suitable for capsule encapsulation, subsequent clinical evaluation, and individualized therapy.

An Insight into the Immunomodulatory Effects of Probiotics in the Prevention of COVID-19 Disease

Abstract: The coronavirus pandemic hit the world with different variants of SARS-CoV-2; reliable therapeutics are needed every hour to control and minimize the infection. To date, the way to menace the chaos of post-COVID infection is not confined rationally. Researchers are still on their way to the progression of an efficient way to eradicate the disease. However, to prevent it from causing infection post-entry into the body, there have been a few strategies to maintain and boost the immune system. At the onset of infection when no antiviral therapeutics were available, convalescent plasma therapies as a proposed mechanism were adapted to treat the post-COVID infection. Researchers have formulated the administration of different types of vaccines based on attenuated or inactivated nucleic acids or subunits after approval from the FDA and still continue to find the best reliable vaccines for better enhancement in inducing immunogenicity of the immune system to fight against the disease. The COVID-19 infection affects the gut and lung axis and there has been dysbiosis of microbiota which leads to cause secondary infections. To accomplish homeostasis of essential microbiota in the body, the administration of different strains of probiotic bacteria has been one way to induce immunogenicity and combat the disease.

BIOTECHNOLOGICAL APPROACHES TO THE CREATION OF NEW FERMENTED DAIRY PRODUCTS

The industrial production of balanced and healthy food has always been one of the most important tasks facing humanity. To solve this problem, research is relevant in the field of creating functional milk-based synbiotic products, which, along with traditional lactic acid microorganisms, include strains of probiotic bacteria with a proven specific positive effect on the macroorganism, and prebiotic substances that increase the selectivity of the benefits of the "useful" microflora of the gastrointestinal tract-intestinal tract and its biological activity.Along with lactic acid bacteria used as starters in fermented milk products, there is a population of predominantly adventitious heterofermentative lactobacilli that can have a positive effect on dairy products.This article describes the use of facultatively and obligately heterofermentative species such as Lactobacillus bulgaricus, Streptococcus thermophilus and Lactobacillus fermentum 14. The general characteristics of these species are described, as well as the desired properties (probiotic activity and taste formation) that affect the characteristics of dairy products.The general characteristics of these species are described along with the desired (probiotic activity and taste formation) properties affecting the characteristics of dairy products.Samples of yogurt based on probiotic starter cultures enriched with plant fillers were studied. The technological process of production was carried out in the traditional way. The dynamics of acid accumulation was studied, titratable and active acidity were determined. As a result of the study, it was revealed that the combination of fermented milk products with plant fillers slightly accelerates the fermentation process.