Probiotic Bacillus Research Articles

Engineering a sustainable protein revolution: Recent advances in cultured meat production

AbstractEmerging as a food of the future, cultured meat is created by growing animal cells outside the organism instead of relying on traditional animal rearing and slaughtering practices. This innovative approach shows great promise in addressing challenges associated with resource utilization, environmental pollution, and public health concerns often encountered in conventional livestock production. In the past decade, popularity of cultured meat has grown enough to be considered as a novel food in regions like Europe, the United Kingdom, and China. This paper discusses the recent advancements in technologies for sustainable cultured meat production. It examines the latest developments in cell sources, cell culture media optimization, bioreactor design, 3D printing, and tissue engineering approaches, which have greatly enhanced the efficiency and scalability of cultured meat production. This food can have a tremendous application as a novel functional food. This review explores the future possibility of applying cultured meat matrix for delivery of hydrophobic nutraceuticals, prebiotics, and probiotics (Bacillus sp).

Transcriptomics reveals the mechanism of high-adhesive Bacillus subtilis inhibiting fresh fish spoilage

Aquatic products are highly susceptible to spoilage caused by adherence of spoilage bacteria during storage, so it is crucial to develop new microecological preservatives. The probiotic B. subtilis can achieve potential preservation effects by adhering to the fish gut to regulate the amount of probiotic colonization in the flora. Here, we demonstrated the surface adhesion characteristics of B. subtilis and the mechanism of adhesion-associated gene-mediated adhesion; the regulation of B. subtilis adhesion ability by nitrogen and carbon sources, and the application of tilapia preservation. Our results revealed 10 B. subtilis adhesion-related genes, among which nitrogen sources up-regulate the adhesion genes hag and motA, carbon sources up-regulate sinR and fliG. Based on flagellum-associated high adhesion strain C6 and high adhesion regulation of B. subtilis C6_H allowed the abundance of probiotic bacteria Bacillus, Rhodobacteraceae to increase, attenuate the content of spoilage flavor substances such as 1-Octen-3-ol, 15-Crown-5 and maintain the freshness of tilapia. The results helped to reveal the adhesion mechanism of B. subtilis and to develop and apply microecological preservatives with high quality adhesion ability.

Effect of Feeding Strategy of Probiotic Bacillus on Improving Water Quality and Growth Performance of Common carp (Cyprinus carpio) Fingerlings

Bacterial probiotics (Bacillus subtilis) and dietary supplements improve the growth and biochemical performance of Common carp (Cyprinus carpio) fingerlings. Fish were nourished for 60 days with a control diet and experimental diets containing B. subtilis 106 to 109 CFU/ ml-l respectively. Dietary supplementation of B. subtilis feeding trial was conducted for 60 days to improve water quality and growth parameters were observed through weight gain, Specific growth rates, and Percent survival of C. carpio fingerlings all through the test period.

Modern probiotics in the treatment of sexually transmitted infections. From the problem to its solution

This article is a review of current scientific publications that are devoted to the problem of sexually transmitted infections (STIs) and the importance of probiotics in their treatment. The article also analyzes the data of regulatory documents of the World Health Organization and statistical data of the Center for Medical Statistics of the Ministry of Health of Ukraine and the State Statistics Service of Ukraine, which highlight the problem of the prevalence of STIs in the world and in Ukraine.Sexually transmitted infections have a serious negative impact on sexual and reproductive health worldwide. In the absence of treatment, they can have serious outcomes, including neurological and cardiovascular diseases, infertility, ectopic pregnancy, increased risk of infection with the human immunodeficiency virus. One of the serious factors which prevent the reduction of STDs increased rate in the world is the drug resistance for these infections. Infections caused by resistant strains of microorganisms are characterized by a more severe course, more often require hospitalization of the patient, increase the duration of the patient’s stay in the hospital, require the use of combined antibiotic therapy with the administration of reserve drugs.According to the results of scientific studies, Bacillus probiotics are an alternative for solving problems related to STIs and their outcomes. The innovative combination of the components of the drugs that contain the SirionProbioProtection probiotic formula helps to overcome such a problem as the resistance of STI pathogens to etiotropic antibiotic therapy. The use of alternative methods of exposure to STIs and their consequences add to success in providing obstetric and gynecological services.

Bacillus licheniformis reduces mortality, improves symptoms, and controls residual pathogenicity in juvenile sea cucumbers Apostichopus japonicus infected with skin ulceration syndrome

Mass mortality caused by skin ulceration syndrome (SUS) is the bottle-neck for the sustainable aquaculture of the sea cucumber Apostichopus japonicus. In the present study, probiotic Bacillus licheniformis (0.25 × 109 CFU/g) was used as the treatment for A. japonicus infected with the SUS that caused by Vibrio harveyi. We found that B. licheniformis significantly reduced the number of infected sea cucumbers 5 days and 7 days after the treatment (group B), compared to those without B. licheniformis treatment (group C) (P < 0.001; P < 0.001). Further, the sea cucumbers fed B. licheniformis had significantly lower mortality at the end of the experiment (<10%) than that of those without the B. licheniformis treatment (>60%) (P < 0.001). These results suggest that the treatment of B. licheniformis is an effective method to reduce the mass mortality resulted from SUS in sea cucumber aquaculture. Further, 3–5 days of treatment significantly improved the adverse symptoms of SUS on the physiology and behavior of sea cucumbers, including the righting behavior, adhesion behavior, food consumption, fecal output and mobility. This indicates B. licheniformis treatment has the advantage in the recovery of sea cucumbers after SUS. Moreover, there was no significant difference observed in the physiology and behavior of sea cucumbers between the SUS infected sea cucumbers after the 7-day treatment of B. licheniformis and the healthy individuals. SUS infected sea cucumbers effectively returned to a stage of normalcy. Further, we found a significantly lower infected rate in sea cucumbers exposed to the culture water of group B (∼5%) than that of those in exposure to the culture water of group C (∼60%). This indicates that the treatment of B. licheniformis efficiently controls the residual pathogenicity of SUS in culture water. The present study demonstrated the effectiveness of B. licheniformis treatment as an environmentally friendly approach to reducing mortality, improving symptoms, and controlling residual pathogenicity in sea cucumber aquaculture.

The Effect of a Bacillus Probiotic and Essential Oils Compared to an Ionophore on the Rumen Microbiome Composition of Feedlot Cattle.

The rising concern of antibiotic growth promoter use in livestock has necessitated the investigation into alternative feed additives. The effect of a probiotic and essential oils to an ionophore on the rumen microbiome composition of Bonsmara bulls raised under feedlot conditions was compared. Forty-eight Bonsmara weaners were allocated to four groups: a group with basal diet (CON) and three groups supplemented with monensin (MON), probiotic (PRO), and essential oils (EO). During the 120 days feeding period, rumen content was collected from four animals per group within each phase via a stomach tube for 16S rRNA and internal transcribed spacer (ITS) sequencing as well as volatile fatty acid analysis. In the starter phase, MON had a significantly lower acetate to propionate ratio and a higher Succinivibrionaceae abundance. The abundance of Lachnospiraceae was significantly higher in EO compared to MON. In the finisher phase, PRO had a significantly higher bacterial diversity. The alpha diversity did not differ between the fungal populations of the groups. The abundance of Proteobacteria was the lowest in PRO compared to the other groups. Limited variation was observed between the rumen microbiome composition of monensin compared to the other treatment groups, indicating that these alternatives can be considered.

Effect of the Probiotic Bacillus subtilis DE-CA9TM on Fecal Scores, Serum Oxidative Stress Markers and Fecal and Serum Metabolome in Healthy Dogs.

There is increasing interest in the use of Bacillus species as probiotics since their spore-forming ability favors their survival in the acidic gastric environment over other probiotic species. The subsequent germination of B. subtilis to their vegetative form allows for their growth in the small intestine and may increase their beneficial effect on the host. B. subtilis strains have also previously been shown to have beneficial effects in humans and production animals, however, no reports are available so far on their use in companion animals. The goal of this study was therefore to investigate the daily administration of 1 × 109 cfu DE-CA9TM orally per day versus placebo on health parameters, fecal scores, fecal microbiome, fecal metabolomics, as well as serum metabolomics and oxidative stress markers in ten healthy Beagle dogs in a parallel, randomized, prospective, placebo-controlled design over a period of 45 days. DE-CA9TM decreased the oxidative status compared to controls for advanced oxidation protein products (AOPP), thiobarbituric acid reactive substances (TBARS) and reactive oxygen metabolites (d-ROMS), suggesting an antioxidant effect of the treatment. Fecal metabolomics revealed a significant reduction in metabolites associated with tryptophan metabolism in the DE-CA9TM-treated group. DE-CA9TM also significantly decreased phenylalanine and homocysteine and increased homoserine and threonine levels. Amino acid metabolism was also affected in the serum metabolome, with increased levels of urea and cadaverine, and reductions in N-acetylornithine in DE-CA9TM compared to controls. Similarly, changes in essential amino acids were observed, with a significant increase in tryptophan and lysine levels and a decrease in homocysteine. An increase in serum guanine and deoxyuridine was also detected, with a decrease in beta-alanine in the animals that ingested DE-CA9TM. Data generated throughout this study suggest that the daily administration of 1 × 109 cfu of DE-CA9TM in healthy Beagle dogs is safe and does not affect markers of general health and fecal scores. Furthermore, DE-CA9TM administration had a potential positive effect on some serum markers of oxidative stress, and protein and lipid metabolism in serum and feces.

Mechanism of TLR4-Mediated Anti-Inflammatory Response Induced by Exopolysaccharide from the Probiotic Bacillus subtilis.

Intestinal inflammatory diseases affect millions of people worldwide, and one class of drugs showing promise toward treatment of several inflammatory diseases is probiotics. Numerous studies have been performed using probiotics to prevent and treat intestinal inflammatory diseases. Most of these studies used intact bacteria, and neither the active molecule nor the molecular mechanisms by which they affect immune responses are known. We have shown that the probiotic Bacillus subtilis is anti-inflammatory and can protect mice from acute colitis induced by the enteric pathogen Citrobacter rodentium. We identified and purified the active molecule, exopolysaccharide (EPS), and showed that it protects mice from C. rodentium-induced colitis by inducing anti-inflammatory M2 macrophages or inhibitory dendritic cells (DCs), both of which inhibit excessive T cell responses. We showed previously that EPS affects macrophages and DCs in a TLR4-dependent manner, and in the current study we asked how EPS induces these anti-inflammatory cells and how they function to inhibit T cells. By investigating the signaling downstream of TLR4 that leads to acquisition of inhibitory properties of macrophages and DCs, we found that EPS induces expression of the inhibitory molecule IDO in bone marrow-derived DCs, and that inhibition of T cell proliferation by IDO-expressing bone marrow-derived DCs utilizes the kynurenine/aryl hydrocarbon receptor circuit. Furthermore, unlike LPS, EPS does not induce inflammatory cytokines upon injection invivo, directly demonstrating different outcomes induced by two different TLR4 agonists.

The Synergic Effect of Gut-Derived Probiotic Bacillus cereus SL1 And Ocimum sanctum on Growth, Intestinal Histopathology, Innate Immunity, and Expression of Enzymatic Antioxidant Genes in Fish, Cirrhinus mrigala (Hamilton, 1822).

An effective alternative approach to combat aquaculture challenges is the strategic application of bioresources, which not only mitigate disease ailment but also optimize fish growth. Hence, current research was undertaken to highlight the synergic role of bioresources such as plant immunostimulant Ocimum sanctum along with potent gut-derived probiotic Bacillus cereus strain SL1 (Gen Bank Accession Number: FJ627945.1) on mrigal (Cirrhinus mrigala) growth, antioxidant status, gut histopathology, and immune response. For 90days, fingerlings (average weight 6.8 ± 0.5g) were fed on diets having varying concentrations of O. sanctum and B. cereus. After the completion of the feeding trial, various growth, immunity, and histological and antioxidant metrics were evaluated according to standard procedures. In comparison to the control and other treatment groups, T3 group showed a significant (P < 0.05) increase in growth parameters, antioxidant enzymatic activity, and hematological and immunological parameters. In addition to it, supplementation of both B. cereus and O. sanctum also upregulated the antioxidant-related gene expressions, such as hepatic catalase gene by 1.89-3.00 folds, hepatic SOD-1 by 4.46-7.52 folds, and GPx-1of the liver by 1.56-1.95 folds. For 10days, fingerlings were challenged with the pathogenic bacterium Aeromonas hydrophila (MTCC-1739), and maximum survival rate (77.77%) was reported in fingerlings of T3 treatment. Further histopathological studies of gut tissues affirm that O. sanctum and B. cereus play a synergic role in the protection of digestive organs from the pathogenic bacterium A. hydrophila. These results suggest that O. sanctum and B. cereus synergically improved the growth performance, immunity, antioxidant status, and gut histology of C. mrigala leading to its sustainable culture.

Male Magelang Duck’s Performance on Grower Period Fed by Probiotics Bacillus amiloliquifaciens And Jengkol Peel Extract

This research was conducted to find out the effect of probiotic bacteria Bacillus amiloliquifaciens and jengkol peel extract (Archidendron jiringa) on the productivity of male Magelang ducks. The treatments in this study were given through water consists of four treatments and five replicants. The treatments in this study were: P0 = Water (without the addition of probiotics and jengkol peel extract, P1 = Water with the addition of 0.5% probiotics + 0.05% jengkol peel extract, P2 = Water with the addition of 1% probiotics + 0.1% jengkol peel extract, P3 = Water with the addition of 1.5% probiotics + 0.15 jengkol peel extract, and P4 = Water with the addition of 2% probiotic + 0.2% jengkol peel extract. Data obtained were analyzed by ANOVA and continued by Duncan’s Multiple Range Test if the results were different. The results showed that the administration of Bacillus amiloliquifaciens and jengkol peel extract had no significant effect on feed intake, water intake, daily weight gain, body weight, and feed conversion of male Magelang ducks (P>0.05). It is concluded that administration of probiotics Bacillus amiloliquifaciens did not improve the performance of male Magelang ducks. Further research is needed on the effectiveness of probiotic bacteria in the intestinal of male Magelang ducks.

Transcriptome analysis of Cryptocaryon irritans tomont responding to Bacillus licheniformis treatment.

Cryptocaryon irritans is a ciliated obligate parasite that causes cryptocaryonosis (white spot disease) and poses great threat to marine fish farming. In recent years, the use of probiotics protects fish from pathogens, which has been identified as the sustainable and environmentally friendly tool to maintain the health and well-being of the host. Accordingly, Cryptocaryon irritans tomont and probiotic Bacillus strain (B.licheniformis, previously isolated from aquaculture water) were co-cultured to detect whether B. licheniformis has anti-C. irritants effect. The result showed that during 4-day incubation, B. licheniformi with 1×107CFU/mL and 1×108CFU/mL concentration effectively inhibited the incubation of C. irritans tomont, indicating that B. licheniformi could inhibit the transformation from reproductive tomont to infective theront of C. irritans. Later, C. irritans samples in the control (without B. licheniformi supplementation) and 1×107CFU/mL B. licheniformi treatment group were sent for transcriptome analysis. Compare with the control group, a total of 3237 differentially expressed genes were identified, among which 626 genes were up-regulated and 2611 genes were down-regulated in 1×107CFU/mL B. licheniformi group. Further Kyoto Encyclopedia of Genes and Genomes pathways analysis showed that anti-C. irritans mechanism of B. licheniformi was mainly involved in the energy metabolism (carbon metabolism, oxidative phosphorylation, biosynthesis of amino acids), transcription and translation (Ribosomes, spliceosomes, RNA transport, etc), lysosome-based degradation (lysosome, phagosome, protein processing in endoplasmic reticulum) and PI3K-Akt pathways. Our study findings raised the possibility of using marine microorganism B. licheniformi in handling aquaculture associated pathogen C. irritans, and preliminarily clarified the molecular mechanism.

Production Optimization and Characterization of Antioxidant Protein Hydrolysate From Piaractus brachypomus Fish Meat By Probiotic Bacillus Strain Isolated From Chicken Gizzard

Production Optimization and Characterization of Antioxidant Protein Hydrolysate From Piaractus brachypomus Fish Meat By Probiotic Bacillus Strain Isolated From Chicken Gizzard

Effects of ribonucleotide supplementation in modulating the growth of probiotic Bacillus subtilis and the synergistic benefits for improving the health performance of Asian seabass (Lates calcarifer).

In aquaculture, due to the requirements for high-density culture, the diseases caused by bacterial pathogens have become a serious issue. To solve this problem, we performed synbiotic application of RNA and Bacillus subtilis as a sustainable and eco-friendly approach to improve the health and immunity of Asian seabass (Lates calcarifer) during cultivation without using any harmful antibiotics or chemicals. Among various forms of nucleic acids, such as mononucleotides and DNA, RNA was found to be most effective in promoting the growth performance of probiotic B. subtilis in all the tested minimal medium conditions. Accordingly, we used the synbiotic combination of B. subtilis and RNA for Asian seabass cultivation. After feed supplementation for fourteen days, the fish that received the combination treatment exhibited a significant increase in innate cellular and humoral immune parameters, including phagocytic activity, phagocytic index, respiratory burst, serum lysozyme and bactericidal activities, as well as upregulated expression of immune-related genes, including HEPC1, A2M, C3, CC, CLEC, LYS, HSP70, and HSP90. Furthermore, significant increases were observed in the ileal villus height and goblet cell numbers in the intestinal villi in all fish treatment groups. The combination treatment did not cause histopathological abnormalities in the intestine and liver, suggesting that the synbiotic treatment is safe for use in fish. The treated Asian seabass also exhibited a significantly increased survival rate after Aeromonas hydrophila challenge. These results indicate that the synbiotic mixture of B. subtilis and RNA can be considered a beneficial feed additive and immunostimulant for Asian seabass cultivation.

Host gut-derived Bacillus probiotics supplementation improves growth performance, serum and liver immunity, gut health, and resistive capacity against Vibrio harveyi infection in hybrid grouper (♀Epinephelus fuscoguttatus×♂Epinephelus lanceolatus).

Several reports have revealed the vital role that probiotics play in fish growth and health. However, few works are available for host gut-derived probiotics on the growth, immunity, and gut microbiota of fish, especially in hybrid grouper (♀Epinephelus fuscoguttatus×♂Epinephelus lanceolatus) due to their isolation difficulty and functional verification. This study aimed at assessing 3 host gut-derived Bacillus species' effects on the growth, immune and antioxidant-biochemical responses, haematological parameters, intestinal morphology, immune-related gene expression, gut microbiota, and disease resistance against Vibrio harveyi in hybrid grouper. A total of 480 hybrid grouper (initial weight=9.03±0.02g) were randomly allotted into 4 groups, namely, the group fed a basal diet without probiotic inclusion (control, B0), the group fed the basal diet with Bacillus velezensis GPSAK4 (BV), the group fed the basal diet with Bacillus subtilis GPSAK9 (BS), and the group fed the basal diet with Bacillus tequilensis GPSAK2 (BT) strains at 1.0×109CFU/g. After a 6-week feeding trial, the results revealed significant improvements (P<0.05) in the growth performance, whole fish-body proximate composition, blood haematological parameters, serum, liver, and intestinal biochemical indexes, intestinal morphology, and protection against V. harveyi pathogen in the probiotic-treated groups compared with the untreated. Additionally, the expressions of intestinal tight junction genes (occludin and ZO1), pro- and anti-inflammatory genes, including IL1β, IL6, IL8, TNFα, MyD88, IL10, and TGFβ, were upregulated (P<0.05) after Bacillus species administration. Host gut-derived Bacillus supplementation shaped the gut microbiota by significantly increasing (P<0.05) the relative abundance of Proteobacteria, Bacteroidetes, Actinobacteria (except the BS group), Acidobacteria (except the BT group), Cyanobacteria (except the BV and BT groups), and Verrucomicrobia phyla, as well as known beneficial genera (Romboutsia, Turicibacter, Epulopiscium, Clostridium_sensu_stricto 1 and 13, Lactobacillus, and Bacillus), but significantly decreased (P<0.05) the abundance of Firmicutes, Chloroflexi, and Fusobacteria phyla, and purported pathogenic genera (Staphylococcus and Photobacterium) compared with the control group. Collectively, the results suggest that B. velezensis GPSAK4, B. subtilis GPSAK9 (especially this strain), B. tequilensis GPSAK2 dietary supplementation at 1.0×109CFU/g has positive effects on the intestinal health of hybrid grouper via microbial composition modulation, thus enhancing the assimilation and absorption of nutrients to boost fish growth, immunity, and disease resistance.

Characterization and anti-inflammatory effect of selenium-enriched probiotic Bacillus amyloliquefaciens C-1, a potential postbiotics

A patented strain of Bacillus amyloliquefaciens C-1 in our laboratory could produce functional sodium selenite (Na2SeO3) under optimized fermentation conditions. With the strong stress resistance and abundant secondary metabolites, C-1 showed potential to be developed as selenium-enriched postbiotics. C-1 has the ability to synthesize SeNPs when incubated with 100 μg/ml Na2SeO3 for 30 h at 30 °C aerobically with 10% seeds-culture. The transformation rate from Na2SeO3 into SeNPs reached to 55.51%. After selenium enrichment, there were no significant morphology changes in C-1 cells but obvious SeNPs accumulated inside of cells, observed by scanning electron microscope and transmission electron microscope, verified by energy dispersive X-ray spectroscopy and X-ray photoelectron spectroscopy. SeNPs had antioxidant activity in radical scavenge of superoxide (O2−), Hydroxyl radical (OH−) and 1,1-diphenyl-2-picryl-hydrazine (DPPH), where scavenging ability of OH− is the highest. Selenium-enriched C-1 had obvious anti-inflammatory effect in protecting integrity of Caco-2 cell membrane destroyed by S. typhimurium; it could preventing inflammatory damage in Caco-2 stressed by 200 μM H2O2 for 4 h, with significantly reduced expression of IL-8 (1.687 vs. 3.487, P = 0.01), IL-1β (1.031 vs. 5.000, P < 0.001), TNF-α (2.677 vs. 9.331, P < 0.001), increased Claudin-1 (0.971 vs. 0.611, P < 0.001) and Occludin (0.750 vs. 0.307, P < 0.001). Transcriptome data analysis showed that there were 381 differential genes in the vegetative growth stage and 1674 differential genes in the sporulation stage of C-1 with and without selenium-enrichment. A total of 22 ABC transporter protein-related genes at vegetative stage and 70 ABC transporter protein-related genes at sporulation stage were founded. Genes encoding MsrA, thiol, glutathione and thioredoxin reduction were significantly up-regulated; genes related to ATP synthase such as atpA and atpD genes showed down-regulated during vegetative stage; the flagellar-related genes (flgG, fliM, fliL, and fliJ) showed down-regulated during sporulation stage. The motility, chemotaxis and colonization ability were weakened along with synthesized SeNPs accumulated intracellular at sporulation stage. B. amyloliquefaciens C-1 could convert extracellular selenite into intracellular SeNPs through the oxidation–reduction pathway, with strong selenium-enriched metabolism. The SeNPs and selenium-enriched cells had potential to be developed as nano-selenium biomaterials and selenium-enriched postbiotics.

Administration of probiotics affects Artemia franciscana metanauplii intestinal ultrastructure and offers resistance against a Photobacterium damselae ssp. piscicida induced oxidative stress response

The effects of Photobacterium damselae ssp. piscicida (Phdp) on immune responses and intestinal ultrastructure of Artemia franciscana following infection and their amelioration by the probiotic bacteria Bacillus subtilis, Lactobacillus plantarum and Lactococcus lactis were evaluated. Pathogen growth inhibition in coculture with each probiotic and its virulence against Artemia were confirmed with an LC50 of 105 CFU mL−1. Phdp administration to Artemia at sublethal levels resulted in depletion of superoxide dismutase, glutathione reductase, glutathione transferase and phenoloxidase activities, extensive lipid peroxidation and reduced survival. Following a combined administration of each probiotic and the pathogen, enzyme activities and survival were significantly higher, while lipid peroxidation was reduced, compared to the infected group with no probiotic treatment (P < 0.05). The transmission electron microscopy study revealed that pathogen infection resulted in disarranged and fragmented microvilli, formation of empty or pathogen containing cytoplasmic vacuoles and damaged mitochondria. In the probiotic-treated and Phdp-infected series, intestinal cells showed normal appearance, except for the presence of pathogen-containing vacuoles and highly ordered but laterally stacked microvilli. The results of the present study indicate that Phdp induces cell death through an oxidative stress response and probiotics enhance Artemia immune responses to protect it against the Phdp induced damage.

Biotransformation of okara (soybean residue) through solid-state fermentation using probiotic Bacillus subtilis and Bacillus coagulans

Okara or soybean residue is a side-stream product of soybean processing generated in large quantities. However, okara is often underutilised as it is prone to spoilage and has a short shelf life. In this study, we explored the feasibility of biotransformation of okara through solid-state fermentation using probiotic Bacillus spp. to modify its nutritional value and flavour property. Two probiotic Bacillus species, namely Bacillus subtilis CU1 (CNCM I-2745) and R0179, and Bacillus coagulans IS-2 and 123 were firstly screened. Further analyses were performed on the B. subtilis R0179 and B. coagulans 123 fermented okara due to their better growth after 24 h (10.28 and 9.24 log colony forming units (CFU)/g respectively). Our results showed that B. subtilis R0179 converted a significant portion of okara insoluble dietary fibre into soluble dietary fibre after fermentation, but not B. coagulans 123. In addition, the samples fermented with B. subtilis R0179 showed higher amounts of maltose, hemicellulosic sugars as well as aromatic, branched-chain and hydrophobic amino acids. Furthermore, the B. subtilis R0179 transformed okara had greater amounts of acetoin, carboxylic acids, pyrazines and branched-chain fatty acids, which may contribute to characteristic natto-like and cheesy flavour. Overall, our study shed light on the possibility of using probiotic bacilli to transform okara with enhanced nutritional value and flavour property, which may increase okara usage in foods.

Development of Nanotechnology-Based Drug Delivery Systems for Controlling Clinical Multidrug-Resistant Staphylococcus aureus and Escherichia coli Associated with Aerobic Vaginitis.

The growing prevalence of resistance to antibiotics potentially makes Escherichia coli and Staphylococcus aureus serious pathogens, necessitating the development of new antimicrobial agents. We extracted crude biosurfactants from a potential probiotic Bacillus spp. to control pathogenic bacteria associated with aerobic vaginal infection. Using nanotechnology formulations, we developed nanoemulsions based on biosurfactants at different concentrations (1% and 3.33%). The results showed that these nanoemulsions were stable, with a weighted index of 0.3, and demonstrated broad-spectrum antibacterial activity against Escherichia coli and Staphylococcus aureus, with MICs ranging between 1.25 and 4 mg/mL. Additionally, the nanoemulsions exhibited interesting antibiofilm effects. All strains became more sensitive to the antibiotics to which they were resistant because of various biosurfactant formulations combined with antibiotics. Lower concentrations of BNE1% and 3.33% were still more efficient than the crude biosurfactants. Our findings demonstrated that the biosurfactant had a strong antibiofilm effect against all tested pathogens. This antibacterial effect can be explained by their ability to alter cell physiology such as cell hydrophobicity and membrane disintegration. Thus, we can conclude that the use of nanotechnology formulations has improved this effect, and the nanoemulsions developed in this study can be used as a potential anti-infectious therapy against multidrug-resistant bacterial strains of clinical origin.

Immunopharmacological aspects of studying immunotropic properties of a novel biocompound

The article presents an experimental study of immunotropic activity of a new compound, a metabiotic, based on metabolites (biologically active substances) produced by a of saprophytic Bacillus subtilis B-9909 strain, being safe for health and standardized at Russian National Collection of Industrial Microorganisms (RNCIM). The aim of our study was to experimentally evaluate the immunotropic effects of metabolites produced by the probiotic Bacillus subtilis B-9909 upon the parameters of cellular immunity tested in the animal model of induced toxic liver damage. The metabolites were isolated from the cultural liquid of Bacillus subtilis culture (RNCIM strain B-9909) during its deep cultivation in a medium containing hydrochloric acid hydrolyzate of soy flour, or pancreatic hydrolyzate of casein. At 16-18 hours of cultivation, the cells were in the exponential growth phase. The indices of cellular status were studied in experimental groups of animals, when assessing therapeutic efficacy of an experimental metabiotic sample, as compared to a group of laboratory animals treated with the reference drug (ursosan). We assessed the quantitative indicators of blood as follows: determination of phagocytic activity (FA) of peripheral blood neutrophils; measurement of metabolic activity in peripheral blood neutrophils by means of NBT-test; quantitative determination of T and B lymphocytes; the number of antibody-forming cells (AFC). Liver damage was studied in a model of acute toxic hepatitis in albino rats. Experimental toxic hepatitis was induced by 40% solution of CCl4 in vaseline oil injected intragastrically for 2 weeks at the dose of 0.2 g/kg. The results of experimental studies showed activation of phagocytic activity by peripheral blood neutrophils at the early terms of experiment, being also confirmed by the results of NBT-testing. Moreover, at the early observation terms, a significantly increased representation of T and B lymphocyte populations as well as AFC numbers, were revealed thus suggesting activation of all components of cellular immunity in response to the carbon tetrachloride toxicity. Thus, the reported studies on the cellular status of laboratory animals treated with metabolites produced by probiotic microorganisms of the genus Bacillus subtilis (B-9909) on laboratory animals with toxic liver damage allow us to conclude that the test sample of this metabiotic preparation exerts a significant immunomodulatory effect, when compared with ursosan. This evidence allows us to consider this compound a promising candidate drug for a new hepatoprotector with immunotropic effect.

Effects of probiotics (Bacillus coagulans) supplementation after antibiotic administration on growth, immunity, and intestinal microflora in turbot Scophthalmus maximus

Effects of probiotics (Bacillus coagulans) supplementation after antibiotic administration on growth, immunity, and intestinal microflora in turbot Scophthalmus maximus