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Spore-forming bacteria and their utilisation as probiotics
Abstract
In this review article, the beneficial application of bacterial spore formers as probiotics in the food industry is discussed based on the knowledge gleaned from current publications. The summary of new scientific results provides evidence of the advantages of the utilisation of Bacillus or Clostridium strains in the food industry. Both bacteria are able to produce a very stable duration form: the endospore. Compared to the widely used lactic acid bacteria, bacterial spores offer the advantage of a higher survival rate during the acidic stomach passage and better stability during the processing and storage of the food product. In many food products, germination of the spores does not occur. Hence the product quality of the food is not affected because of their inactive metabolism. Besides the possible utilisation and functional properties, an overview of the fast-developing knowledge about the mechanisms of the beneficial health effects of spore-forming bacteria is provided.
... In this same study (2), there was a decrease in the abundance of beneficial microbes, such as Bifidobacterium, Bacteriodetes, Lactobacillus and Clostridaceae. Importantly, Bifidobacterium and Lactobacillus are commonly used as probiotics with the intention of restoring a healthy GI microbial state, while Clostridaceae is a family of spore forming bacteria (SFB) that has also shown beneficial properties (4). Bacteriodetes have also been found to exhibit a mutualistic relationship within the GI tract of their human hosts (5). ...
... In the present study, we interrogated changes in several core members of the GI microbiota that have been previously linked to perturbations to the gut-brain axis (SFB, Bifidobacterium, Lactobacillus and Akkermansia) after exposure to 4 He radiation, as well as alterations in the GI tract and neurobehavioral performance. We report alterations to these core components as well as deficits in social odor recognition memory after 4 He exposure. ...
... Consequently, any behavioral testing performed, or samples collected within these 10 days occurred at BNL (Fig. 1). Rats were placed in individual plastic holders, and then acutely exposed to 4 He ions (250 MeV/n) generated at the NASA Space Radiation Laboratory (NSRL) facility at BNL (sham: n ¼ 25; 5 cGy: n ¼ 29; and 25 cGy: n ¼ 29). 4 He ions of this energy have a range in water of 37.6 cm with an average linear energy transfer (LET) of 1.58 keV/lm. ...
Beyond low-Earth orbit, space radiation poses significant risks to astronaut health. Previous studies have shown that the microbial composition of the gastrointestinal (GI) microbiome changes upon exposure to high-linear energy transfer radiation. Interestingly, radiation-induced shifts in GI microbiota composition are linked to various neuropsychological disorders. Herein, we aimed to study changes in GI microbiota and behaviors of rats exposed to whole-body radiation (0, 5 or 25 cGy 4He, 250 MeV/n) at approximately 6 months of age. Fecal samples were collected 24 h prior to 4He irradiation and 24 h and 7 days postirradiation for quantitative PCR analyses to assess fecal levels of spore-forming bacteria (SFB), Bifidobacterium, Lactobacillus and Akkermansia. Rats were also tested in the social odor recognition memory (SORM) test at day 7 after 4He exposure. A subset of rats was euthanized 90 min after completion of the SORM test, and GI tissue from small intestine to colon were prepared for examining overall histological changes and immunohistochemical staining for serotonin (5-HT). No notable pathological changes were observed in GI tissues. Akkermansia spp. and SFB were significantly decreased in the 25 cGy group at 24 h and 7 days postirradiation compared to pre-exposure, respectively. Bifidobacterium and Lactobacillus spp. showed no significant changes. 5-HT production was significantly higher in the proximal small intestine and the cecum in the 25 cGy group compared to the sham group. The 25 cGy group exhibited deficits in recognition in SORM testing at day 7 postirradiation. Taken together, these results suggest a connection between GI microbiome composition, serotonin production, and neurobehavioral performance, and that this connection may be disrupted upon exposure to 25 cGy of 4He ions.
... B. coagulans has a number of physiological characteristics that distinguish it from other probiotic Bacillus species, including growth conditions, biochemical reactions, the use of carbon sources, and a cell wall with high lipid contents compared to other Gram-positive bacteria [9,10]. Compared to non-spore-forming probiotic bacteria such as Lactobacillus spp., B. coagulans offers the advantage of improved stability during industrial processing and storage in functional foods [11,12]. The ability to withstand stomach acid and bile salts provides B. coagulans an edge for longer survival periods in the gastrointestinal tract [13][14][15][16]. ...
... Microorganisms 2023,11, 2564 ...
Spore-forming probiotic bacteria, including Bacillus coagulans, are resilient and produce a variety of beneficial metabolites. We evaluated the immune-modulating effects of the novel probiotic strain Bacillus coagulans JBI-YZ6.3, where the germinated spores, metabolite fraction, and cell wall fraction were tested in parallel using human peripheral blood mononuclear cell cultures under both normal and lipopolysaccharide-induced inflamed culture conditions. The expression of CD25 and CD69 activation markers was evaluated via flow cytometry. Supernatants were tested for cytokines, interferons, chemokines, and growth factors using Luminex arrays. The germinated spores were highly immunogenic; both the cell wall and metabolite fractions contributed significantly. Under normal culture conditions, increased levels of immune activation were observed as increased expressions of CD25 and CD69 relative to natural killer cells, suggesting an increased ability to attack virus-infected target cells. On monocytes, a complex effect was observed, where the expression of CD25 increased under normal conditions but decreased under inflamed conditions. This, in combination with increased interleukin-10 (IL-10) and decreased monocyte chemoattractant protein-1 (MCP-1) production under inflamed conditions, points to anti-inflammatory effects. The production of the stem cell-related growth factor granulocyte colony-stimulating Factor (G-CSF) was enhanced. Further research is warranted to characterize the composition of the postbiotic metabolite fraction and document the characteristics of immunomodulating agents secreted by this probiotic strain.
... Bacillus species have been used as probiotics for at least 50 years and are commonly employed as growth promoters and competitive exclusion agents in animal production [9]. This is attributed to their high-temperature resistance [10,11], acid and alkali resistance [12,13], ease of culture, storage stability, and low processing loss [13][14][15]. Amongst these species, Bacillus subtilis is a typical representative; its subordinate strain, Bacillus amyloliquefaciens, can secrete various protein-active substances. ...
... Bacillus species have been used as probiotics for at least 50 years and are commonly employed as growth promoters and competitive exclusion agents in animal production [9]. This is attributed to their high-temperature resistance [10,11], acid and alkali resistance [12,13], ease of culture, storage stability, and low processing loss [13][14][15]. Amongst these species, Bacillus subtilis is a typical representative; its subordinate strain, Bacillus amyloliquefaciens, can secrete various protein-active substances. The gryA gene is an important taxonomic basis of Bacillus amyloliquefaciens [16,17]. ...
[Background] Bacillus LFB112 is a strain of Bacillus amyloliquefaciens screened in our laboratory. Previous studies found that it has a strong ability for fatty acid metabolism and can improve the lipid metabolism of broilers when used as feed additives. [Methods] This study aimed to confirm the fatty acid metabolism of Bacillus LFB112. Sterilized soybean oil (SSO) was added to the Beef Peptone Yeast (BPY) medium, and its effect on fatty acid content in the supernatant and bacteria, as well as expression levels of genes related to fatty acid metabolism, were studied. The control group was the original culture medium without oil. [Results] Acetic acid produced by the SSO group of Bacillus LFB112 decreased, but the content of unsaturated fatty acids increased. The 1.6% SSO group significantly increased the contents of pyruvate and acetyl-CoA in the pellets. Furthermore, the mRNA levels of enzymes involved in the type II fatty acid synthesis pathway of FabD, FabH, FabG, FabZ, FabI, and FabF were up-regulated. [Conclusions] Soybean oil increased the content of acetyl-CoA in Bacillus LFB112, activated its type II fatty acid synthesis pathway, and improved the fatty acid metabolism level of Bacillus LFB112. These intriguing results pave the way for further investigations into the intricate interplay between Bacillus LFB112 and fatty acid metabolism, with potential applications in animal nutrition and feed additive development.
... Among these probiotics, microbes of the Bacillus species represent an ever-growing supplement in human studies (Acosta-Rodriguez-Bueno et al., 2022;Anaya-Loyola et al., 2019;Bader et al., 2012;Elshaghabee et al., 2017;Goldenberg et al., 2015;Guo et al., 2022). Bacillus spp. ...
... The use of Bacillus probiotics in maintenance of gut health has been largely supported in the last years and has driven its clinical applications (Acosta- Rodriguez-Bueno et al., 2022;Bader et al., 2012;Elshaghabee et al., 2017;Goldenberg et al., 2015;Nicholson, 2002;Wong-Chew et al., 2022). Their favourable effects have been linked to several properties, such as antimicrobial and immunomodulatory activity, regulation of cell growth and differentiation, cellcell signalling, cell adhesion, signal transcription and transduction, production of vitamins and gut protection from genotoxic agents. ...
In the present study, the safety, tolerance and impact of 1×109 cfu Bacillus clausii CSI08, 1×109 cfu Bacillus megaterium MIT411 and a probiotic cocktail containing Bacillus subtilis DE111®, Bacillus megaterium MIT411, Bacillus coagulans CGI314, and Bacillus clausii CSI08 with a total count of 2.0×109 cfu administered daily were assessed as compared with a maltodextrin containing placebo control. A total of 98 study participants received daily doses for 45 days, followed by a washout period of 2 weeks. A questionnaire to capture the incidence and duration of upper respiratory tract, urinary tract and/or gastrointestinal complaints and a diary to capture stool regularity and consistency was kept daily to record compliance throughout the 45 days. Faecal and blood samples were collected for microbiological and haematological analysis at the start and end of the treatment period. The probiotic cocktail significantly decreased the incidence of loose stools throughout the entire study. The recorded respiratory, urinary and gastrointestinal symptoms, defecation frequency and other stool consistency were not influenced. No clinically relevant changes in blood parameters, such as liver and kidney function and no serious adverse events appeared during and after administration. There were no changes in symptoms including sadness, irritability, energy, appetite, tension, stress, sleep, cardiovascular events, aches and pains, and dizziness as determined by a mood questionnaire administered to participants at baseline and at the end of the treatment period. Similarly, the measured inflammatory cytokines, antioxidant levels, cholesterol, triglycerides, free amino acids or minerals remained unaffected. There were no negative changes in alpha or beta diversity of the microbiota with any of the treatment groups. These promising data suggest that these treatments were safe and well tolerated, and further work with larger cohorts are justified to determine the efficacy of these potential probiotics in select demographic groups. Trial registration number with clinicaltrials.gov at NCT04758845.
... Of these six candidates, only Lysinibacillus was a spore former. This characteristic might benefit the colonization of the gut, since these endospores have a higher survival rate through the acidic gut passage [60]. In fact, probiotics must pass the lumen of the middle midgut with a low pH around 2 and high lysozyme activity to arrive in the posterior midgut with a pH of approximately 8.5 [61]. ...
... Further, inoculating the substrate on a more frequent basis (for instance daily as executed by [40], but not discussed by the authors) could probably enhance gut colonization more than a one-off inoculation [14], though it also increases the risk of observing nutritional effects of the continuous supplementation. As already discussed in Section 7.1, the form in which the probiotic is offered to the larvae can affect the outcome of the inoculation trial, and it might be advised to use lyophilized or protected cells or if possible spores, which are more likely to persist the passage through the middle midgut [14,60,70]. Finally, for multi-bacterial inoculants, the outcomes were dependent on the candidates and on the ratio of the candidates [38,42]. ...
Given the novelty of the industrial production of the edible insects sector, research has primarily focused on the zootechnical performances of black soldier fly larvae (BSFL) in response to different substrates and rearing conditions as a basis to optimize yield and quality. However recently, research has started to focus more on the associated microbes in the larval digestive system and their substrates and the effect of manipulating the composition of these communities on insect performance as a form of microbiome engineering. Here we present an overview of the existing literature on the use of microorganisms during rearing of the BSFL to optimize the productivity of this insect. These studies have had variable outcomes and potential explanations for this variation are offered to inspire future research that might lead to a better success rate for microbiome engineering in BSFL.
... Bacillus spp. are the most commonly used probiotic microorganisms because of their ability to form endospores [6]. This enables them to survive the feed manufacturing process and the passage through the stomach. ...
... This enables them to survive the feed manufacturing process and the passage through the stomach. Moreover, spores allow easy administration, storage and prolonged shelf-life [6]. One frequently used species, Bacillus subtilis, is considered to be safe for consumption [7,8]. ...
Simple Summary
Bacterial strains that are consumed by humans or by animals to promote health are called probiotics. In poultry, Bacillus strains are widely used as feed additives for this purpose. Although different modes of action have been proposed, studies showing effects on what metabolites the bacteria produce in a test tube, and whether these can also be found in the intestine of animals that were given these strains as feed additives, are lacking. In the current study, we show that administration of a Bacillus strain to broiler chickens changes the microbial composition in the gut by reducing opportunistic pathogenic bacterial families and promoting beneficial bacterial families. We show that two molecules, hypoxanthine and nicotinic acid, are produced by the Bacillus strain and are elevated in the intestinal tract of these animals. We hypothesize that nicotinic acid can be used by beneficial microbes and is essential for their intestinal colonization, and that both molecules can have a positive effect on the intestinal wall. These data can be used to evaluate and develop novel feed additives to promote health of chickens, and reduce the need for antibiotic usage.
Abstract
The probiotic Bacillus subtilis strain 29784 (Bs29784) has been shown to improve performance in broilers. In this study, we used a metabolomic and 16S rRNA gene sequencing approach to evaluate effects of Bs29874 in the broiler intestine. Nicotinic acid and hypoxanthine were key metabolites that were produced by the strain in vitro and were also found in vivo to be increased in small intestinal content of broilers fed Bs29784 as dietary additive. Both metabolites have well-described anti-inflammatory effects in the intestine. Furthermore, Bs29784 supplementation to the feed significantly altered the ileal microbiome of 13-day-old broilers, thereby increasing the abundance of genus Bacillus, while decreasing genera and OTUs belonging to the Lactobacillaceae and Enterobacteriacae families. Moreover, Bs29784 did not change the cecal microbial community structure, but specifically enriched members of the family Clostridiales VadinBB60, as well as the butyrate-producing families Ruminococcaceae and Lachnospiraceae. The abundance of various OTUs and genera belonging to these families was significantly associated with nicotinic acid levels in the cecum, suggesting a possible cross-feeding between B. subtilis strain 29784 and these beneficial microbes. Taken together, the data indicate that Bs29784 exerts its described probiotic effects through a combined action of its metabolites on both the host and its microbiome.
... However, the market size of Bacillus-based products should increase significantly as demand is enhanced! Bacillus spores offer the advantage of a higher survival rate during acidic stomach passage, easy multiplication and maintained viability in the GIT, and improved stability during the processing and storage of the probiotic [23,25]. ...
... This review does not intend to itemize all the sporeforming bacteria or all Bacillus sp. probiotic actions; for a more detailed description, refer to Bader et al. [25], Permpoonpattana et al. [34], Elshaghabee et al. [14], Bernardeau et al. [35], and Mingmongkolchai and Panbangred [36]. ...
The growing global demand for animal products and processed meat has created a challenge for the livestock sector to enhance animal productivity without compromising product quality. The restriction of antibiotics in animal feeds as growth promoters makes the use of probiotics a natural and safe alternative to obtain functional foods that provide animal health and quality and to maintain food safety for consumers. To incorporate these additives into the diet, detailed studies are required, in which in vitro and in vivo assays are used to prove the efficacy and to ensure the safety of probiotic candidate strains. Studies on the use of Bacillus subtilis natto as a spore-forming probiotic bacterium in animal nutrition have shown no hazardous effects and have demonstrated the effectiveness of its use as a probiotic, mainly due to its proven antimicrobial, anti-inflammatory, antioxidant, enzymatic, and immunomodulatory activity. This review summarizes the recent scientific background on the probiotic effects of B. subtilis natto in animal nutrition. It focuses on its safety assessment, host-associated efficacy, and industrial requirements.
... Metabolites characteristic has contributed to the massive potential of antibiotics in multiple fields of agriculture, industry, and medications (Bahaddad et al., 2023;Danilova and Sharipova, 2020). These species are considered safe and efficacious probiotics, as they do not produce toxic substances and are manageable in terms of their growth requirements and tolerance to high temperatures (Saggese et al., 2021), in addition to classic probiotic traits, that are characterized by high stability towards processing and storage (Bader et al., 2012), and at low production costs. Several modes of action mediate its beneficial effects) (de Simone, 2019; Wieërs et al., 2020). ...
... Metabolites characteristic has contributed to the massive potential of antibiotics in multiple fields of agriculture, industry, and medications (Bahaddad et al., 2023;Danilova and Sharipova, 2020). These species are considered safe and efficacious probiotics, as they do not produce toxic substances and are manageable in terms of their growth requirements and tolerance to high temperatures (Saggese et al., 2021), in addition to classic probiotic traits, that are characterized by high stability towards processing and storage (Bader et al., 2012), and at low production costs. Several modes of action mediate its beneficial effects) (de Simone, 2019;Wieërs et al., 2020). ...
... Bacillus species are renowned for their remarkable ability to withstand extreme environmental conditions, such as acids and bile salts in the gastrointestinal tracts, heat processing, and low-temperature storage [146,147]. This resilience is largely due to their capacity to form endospores, a complex developmental process where the bacterial cells differentiate into resistant spores, allowing their survival in extreme temperatures, variations in pH, ultraviolet radiations, harmful chemicals, and nutrient shortage [148]. ...
Mycotoxins are toxic secondary metabolites produced by various types of fungi that are known to contaminate various food products; their presence in the food chain poses significant risks to human and animal health and leads to enormous economic losses in the food and feed industry worldwide. Ensuring food safety and quality by detoxifying mycotoxin is therefore of paramount importance. Several procedures to control fungal toxins have been extensively investigated, such as preventive measures, physical and chemical methods, and biological strategies. In recent years, microbial degradation of mycotoxins has attracted much attention due to its reliability, efficiency, and cost-effectiveness. Notably, bacterial species from the Bacillus genus have emerged as promising candidates for mycotoxin decontamination owing to their diverse metabolic capabilities and resilience in harsh environmental conditions. This review manuscript aims to provide a summary of recent studies on the biodegradation of fungal toxins by Bacillus bacteria, thereby illustrating their potential applications in the development of mycotoxin-degrading products.
... Bacillus spp. generally are advantageous compared to commonly used probiotics such as lactic acid bacteria, largely due to their capacity to form spores. Bacterial spores offer high survival rates, enhanced tolerance to stress, and stability during processing and storage of product (Bader, Albin, and Stahl 2012;Elshaghabee et al. 2017). When incorporated onto dry feeds, metabolically inactive spores do not germinate for months or longer, which contributes to the extended shelf life (Wang, Li, and Lin 2008). ...
Early mortality syndrome (EMS) or acute hepatopancreatic necrosis syndrome (AHPND) is an epizootic bacterial infection of shrimp that has had significant negative impact on the global shrimp aquaculture industry. This disease is largely attributed to pathogenic Vibrio parahaemolyticus EMS strains (VP‐EMS). Application of probiotics was examined for control of pathogenic VP‐EMS infections in Pacific white shrimp (Litopenaeus vannamei) under laboratory‐scale conditions. Two probiotic products, a single strain (O14VRQ) of Bacillus subtilis and a blend (Plus10) of five strains of Bacillus from four different species (Bacillus amyloliquifaciens, Bacillus megaterium, Bacillus brevis, and two distinct strains of Bacillus subtilis), were evaluated at various concentrations as feed additives (applied as a top coat on commercial feed) or dosed directly into the culture water. Two trials were conducted in which shrimp were fed either a control feed (no probiotics) or probiotic‐coated feed for 7 days prior to be being challenged with VP‐EMS. Shrimp were observed for clinical signs of disease and mortalities during the disease challenge study. These experiments demonstrated that while both Bacillus probiotic products were shown to significantly (p < 0.05) improve shrimp survival, the O14VRQ strain provided the most consistent protection across the trials in top‐coated feed. Overall, better efficacy was observed with probiotic‐coated feeds. Findings were directly and positively correlated with concentration for top‐coated and even more so for direct in‐tank applications. Collectively, these findings contribute to our understanding about how probiotic strains could be applied to enhance shrimp health in aquaculture.
... Moreover, the acid tolerance of these isolates could be due to their origin and H+-ATPase activity (9). Further, the tolerance to stressful and acidic conditions signposts them as suitable candidates for probiotic applications (42). For probiotics the autochthonous bacteria isolated from the same host are preferred for being more compatible with the intestinal microbiota and prevailing micro-environmental conditions (9). ...
Background and Objectives
Rumen microbiologists are looking for new probiotics to improve the digestibility of livestock diets. This study intended to screen and evaluate the ruminal cellulolytic bacteria (CBs) and their potential application as probiotics.
Materials and Methods
Microbial culture and molecular techniques performed to isolate CBs from the rumen of camels, deer and rams. Their antibacterial and antibiogram tests were done using disc diffusion method. Their potential to degrade cellulose, starch, tannin and protein were investigated using clear zone halo, and spectrophotometric techniques. Bilious, saline, and acidic broth media were used to study the resistance of isolates in intestinal conditions.
Results
The phylogenetic analysis revealed that the strains belonged to Firmicutes and Proteobacteria phyla, Citrobacter murliniae, Ornithinibacillus bavariensis, C. braakii, and Bacillus subtilis. The highest cellulase (CAS) activity was recorded by C. murliniae Dez wildlife13A (2.98 UmL⁻¹), whereas C. braakii Loot desert 111A (1.14 Uml⁻¹) was produced the lowest enzyme. The isolates were highly resistant to synthetic conditions of intestine (pH 2.5–3.5, bile 0.3–2%), as well as tolerated higher concentrations of NaCl (up to 10%). They effectively inhibited standard pathogen strains, and showed sensitivity to the used antibiotics.
Conclusion
This study reports the cellulolytic O. bavariensis Tabbas desert 32A for the first time from the rumen, which will have potential biotechnological applications.
... The most well-known probiotic bacteria are lactic acidproducing Lactobacillus and Bifidobacterium organisms. Spore-forming Bacillus probiotics have attracted more attention in recent years due to their intrinsic ability to survive harsh industrial processing conditions (thermal processing and low moisture) and passage through the gastrointestinal tract (low pH and bile salts) [12,13]. Specifically, Bacillus coagulans exhibits characteristics of both Bacillus and Lactobacillus and has been developed into various commercial products (BC30, LactoSpore, Sporlac, LACBON). ...
Spore-forming Bacillus coagulans has been widely recognized as an important probiotic, which is commonly used in products for human consumption and animal feeds. B. coagulans exhibits beneficial traits from both Bacillus and lactic acid-producing Lactobacillus. The present study evaluated the safety of the newly isolated B. coagulans strain JBI-YZ6.3, using combined genomic and phenotypic analysis approaches. The taxonomic classification based on genome sequence and biochemical tests identified strain JBI-YZ6.3 as B. coagulans. Comprehensive genome-based analyses established JBI-YZ6.3 as a novel strain of B. coagulans. Antibiotic susceptibility testing showed that the strain JBI-YZ6.3 was sensitive to a panel of fourteen antibiotics, and no genes related to antibiotic resistance were found in its genome. The spores of strain JBI-YZ6.3 exhibited tolerance to acid and bile salts, as well as stability at ambient and elevated conditions of temperature and relative humidity. There were no homologs of Bacillus toxin genes identified in the genome of JBI-YZ6.3, and the strain exhibited no cytotoxicity towards Vero cells and human peripheral blood mononuclear cells. In conclusion, findings from this study support the safety of B. coagulans strain JBI-YZ6.3, which can be developed into new probiotic products for preventive and therapeutic benefits in human and animal hosts.
... Moreover, the acid tolerance of these isolates could be due to their origin and H+-ATPase activity (9). Further, the tolerance to stressful and acidic conditions signposts them as suitable candidates for probiotic applications (42). For probiotics the autochthonous bacteria isolated from the same host are preferred for being more compatible with the intestinal microbiota and prevailing micro-environmental conditions (9). ...
Background and Objectives: Rumen microbiologists are looking for new probiotics to improve livestock diets. This study intended to screen and evaluate the ruminal cellulolytic bacteria (CBs) and their potential application as probiotics. Materials and Methods: Microbial culture and molecular techniques performed to isolate CBs from the rumen of camels, deer and rams. Their antibacterial and antibiogram tests were done using disc diffusion method. Their potential to degrade cellulose, starch, tannin and protein were investigated using clear zone halo, and spectrophotometric techniques. Bilious, saline, and acidic broth media were used to study the resistance of isolates in intestinal conditions. Results: The phylogenetic analysis revealed that the strains belonged to Firmicutes and Proteobacteria phyla, Citrobacter murliniae, Ornithinibacillus bavariensis, C. braakii, and Bacillus subtilis. The highest cellulase (CAS) activity was recorded by C. murliniae Dez wildlife13A (2.98 UmL-1), whereas C. braakii Loot desert 111A (1.14 Uml-1) was produced the lowest enzyme. The isolates were highly resistant to synthetic conditions of intestine (pH 2.5-3.5, bile and 0.3-2%), as well as tolerated higher concentrations of NaCl (up to 10%). They effectively inhibited standard pathogen strains, and showed sensitivity to the used antibiotics. Conclusion: This study reports the cellulolytic O. bavariensis Tabbas desert 32A for the first time from the rumen, which will have potential biotechnological applications.
... The results for E. faecium IDCC 2102 indicated an approximately 80% viability with increased omega-3 fatty acids concentrations under acidic stress, whereas that of B. coagulans IDCC 1201 decreased with increased omega-3 fatty acids concentrations from 100% to 80% when compared to the control (0%) (Figure 3). The viability of B. coagulans IDCC 1201 was superior to that of E. faecium IDCC 2102, possibly due to spore formation observed in the B. coagulans strain (1 × 10 8 CFU·mL -1 ) were inoculated in acidic medium at 37 °C at 3 h; the acidic medium adjusted pH 2.5 and treated omega-3 fatty acids with concentrations of 0, 0.5, 1, and 2%; the standard error (n = 3 independent experiments) is indicated in the error bar; CFU -colony forming unit (Bader et al. 2012). The bile tolerance test showed that omega-3 fatty acids supplementation did not affect viability at 0.3% (w/v) bile acid when compared to the control (0%) (Figure 4). ...
... It was found that B. cereus and C. perfringens existed in approximately a third of the tested hundred vegetable samples, which were mostly in the form of salads, obtained from United Kingdom retail stores. Only when the produce is processed in a manner that permits the growth of spores and the growth of vegetative cells is there a threat to public health posed by these spore-forming bacteria (Bader et al., 2012). ...
... The observed anti-diabetic effects are due to the modulation of gut microbiota composition, regulation of immune reactions, and improvement of energy metabolism [9]. Spores-based probiotics have additional benefits, such as higher resistance to gastric acid and increased stability at room temperature [10]. MSB, a probiotic mixture of five spore-forming Bacillus strains, has enriched the microbial diversity in an in vitro model of a simulator of the human intestinal microbial ecosystem (SHIME ® ) and ameliorated the production of short-chain fatty acids [11]. ...
Glucocorticoids are effective anti-inflammatory and immunosuppressive agents. Long-term exposure is associated with multiple metabolic side effects. Spore-forming probiotic bacteria have shown modulatory properties regarding glycolipid metabolism and inflammation. The aim of this study was to evaluate, for the first time, the effects of Bacillus species spores (B. licheniformis, B. indicus, B. subtilis, B. clausii, and B. coagulans) alone and in combination with metformin against dexamethasone-induced systemic disturbances. A total of 30 rats were randomly divided into 5 groups: group 1 served as control (CONTROL), group 2 received dexamethasone (DEXA), group 3 received DEXA and MegaSporeBiotic (MSB), group 4 received DEXA and metformin (MET), and group 5 received DEXA, MSB, and MET. On the last day of the experiment, blood samples and liver tissue samples for histopathological examination were collected. We determined serum glucose, total cholesterol, triglycerides, tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), interleukin-10 (IL-10), catalase, total antioxidant capacity (TAC), and metformin concentration. DEXA administration caused hyperglycemia and hyperlipidemia, increased inflammation cytokines, and decreased antioxidant markers. Treatment with MSB reduced total cholesterol, suggesting that the administration of Bacillus spores-based probiotics to DEXA-treated rats could ameliorate metabolic parameters.
... The high temperature and humidity of the storage conditions can induce lethal injury that promotes loss of cell survival in freeze-and spray-dried probiotics throughout the storage period [13]. Bacillus subtilis KKU213 appears to be more tolerant during storage under cold and ambient air temperatures due to its ability to produce an endospore, which may lead to higher stability and survival rates during storage [24]. In addition, the low k values of freeze-and spray-dried B. subtilis KKU213 also support the notion that spore-forming B. subtilis exhibits higher storage stability. ...
Background and Aim
Many strains of probiotics have been exploited and used as animal dietary supplements for broiler production. The efficacy and survival of probiotics during production may reflect better activities of the probiotics in the host. This study investigated the effects of freeze- and spray-drying on the survivability and properties of probiotics and their ability to improve the growth and health performance of broilers.
Materials and Methods
Probiotic powders of four strains of lactic acid bacteria, Enterococcus faecium CA4, Enterococcus durans CH33, Ligilactobacillus salivarius CH24, Pediococcus acidilactici SH8, and Bacillus subtilis KKU213, were prepared using rice bran/chitosan/carboxy methyl cellulose as the carrier. The survival of each probiotic strain was investigated under stress conditions, including freeze-drying, spray-drying, and simulated gastrointestinal conditions. The body weight gain (BWG) and intestinal histomorphology were determined to assess broiler growth performance.
Results
All dried probiotics yielded a high survival rate during freeze-drying (95.8–98.6%) and spray-drying (94.4–98.2%). In addition, an analysis of the main effect revealed that the effectiveness of freeze-drying was higher than that of spray-drying in minimizing the loss of cell viability. The antimicrobial activity of all immobilized dried probiotic strains against Salmonella was maintained. The immobilized probiotics tolerated a low pH value of 2.0 and 0.5% (w/v) bile salt. Probiotic administration of a mixture of the five dried probiotics to 1-day-old hatched male broilers at early and late ages resulted in potential colonization in the broiler intestine, and enhancements in the BWG, lipid metabolism, and gut health (villus height and cryptal depth) were observed in the probiotic-treated groups.
Conclusion
The administration of three doses of the spray-dried probiotic mixture at days 15, 17, and 19 after hatching was sufficient to achieve long-term growth and health benefits in broilers. This finding might provide a cost-effective alternative to the administration of commonly used antibiotics in broiler production.
... Dormant bacterial spores are extremely resistant and can cope with the toxic effects of atmospheric oxygen, ultraviolet radiation, lack of nutrients, adverse temperatures, and desiccation (Browne et al., 2017). Bacterial spores have great application potential as probiotic food supplements, as they offer the benefit of higher survival rates during passage through the acidic stomach and better stability during processing and storage than the widely used lactic acid bacteria (Bader et al., 2012). ...
The host–intestinal microbiome interaction has gained much scientific attention in the past two decades, boosted by advances in DNA sequencing and cultivation techniques. An accumulating amount of evidence shows that gut microbes play crucial roles in gut homeostasis, immune system education, and are associated with quality-of-life indicators. Beneficial health factors are associated with the digestion of dietary fibres in the colon and the subsequent production of short-chain fatty acids, including acetate, propionate, and butyrate. Coprococcus is a butyrate-producing genus in the phylum Firmicutes, and its abundance is inversely correlated with several neuropsychological and neurodegenerative disorders. Case–control studies provide strong evidence of decreased abundance of Coprococcus spp. in depressed individuals. The species Coprococcus eutactus has the unique capacity to use two separate pathways for butyrate synthesis and has been found to be depleted in children with delayed language development and adults with Parkinson’s disease. The combined literature on Coprococcus and the gut microbiota–brain axis points towards enhanced butyrate production and reduced colonisation of pathogenic clades as factors explaining its association with health effects. The genus Coprococcus is a promising candidate for a mental health biomarker and an interesting lead for novel dietary-based preventive therapies for specific neurological disorders.
... Probiotic supplementation could represent a novel therapeutic strategy in the treatment of gut microbiota dysbiosis in patients with psoriasis, as supplementation has been associated with significant alleviation of psoriasis-like pathogenic characteristics and a reduced proinflammatory status, in both experimental and clinical trials [12][13][14]. Sporeforming Bacillus probiotics have several advantages over non-spore-forming probiotics, as they can tolerate room temperature storage and are more resistant to stomach acids [15]. As such, they survive the harsh conditions of gastric passage, entering into the intestines completely viable, where they are able to germinate in high numbers [16,17]. ...
Psoriasis is a chronic inflammatory skin disease with autoimmune pathological characteristics. Recent research has found a link between psoriasis, inflammation, and gut microbiota dysbiosis, and that probiotics and prebiotics provide benefits to patients. This 12-week open-label, single-center clinical trial evaluated the efficacy of probiotics (Bacillus indicus (HU36), Bacillus subtilis (HU58), Bacillus coagulans (SC208), Bacillus licheniformis (SL307), and Bacillus clausii (SC109)) and precision prebiotics (fructooligosaccharides, xylooligosaccharides, and galactooligosaccharides) in patients with psoriasis receiving topical therapy, with an emphasis on potential metabolic, immunological, and gut microbiota changes. In total, 63 patients were evaluated, with the first 42 enrolled patients assigned to the intervention group and the next 21 assigned to the control group (2:1 ratio; non-randomized). There were between-group differences in several patient characteristics at baseline, including age, psoriasis severity (the incidence of severe psoriasis was greater in the intervention group than in the control group), the presence of nail psoriasis, and psoriatic arthritis, though it is not clear whether or how these differences may have affected the study findings. Patients with psoriasis receiving anti-psoriatic local therapy and probiotic and prebiotic supplementation performed better in measures of disease activity, including Psoriasis Area and Severity Index, Dermatology Life Quality Index, inflammatory markers, and skin thickness compared with those not receiving supplementation. Furthermore, in the 15/42 patients in the intervention group who received gut microbiota analysis, the gut microbiota changed favorably following 12 weeks of probiotic and prebiotic supplementation, with a shift towards an anti-inflammatory profile.
... Unfortunately, little is known about the actual mechanisms through which these bacteria produce their therapeutic effects. Studies have shown that the use of spore-forming probiotics in the place of antibiotics leads to strengthen immune system and improve animal growth (Bader et al., 2012;Cutting, 2011;Shiravamaiah et al., 2011). In a study conducted by Permpoonpattana et al. (2012), the authors suggested that spore-forming probiotics administered to mice are able to suppress infectious agents like Escherichia coli, Clostridium perfringens, and Salmonella enteritidis. ...
Background: Higher resistance of microbial spores to high temperatures, low pH and high pressures coupled with characteristics such as good stability and rapid germination make their use extremely desirable for the innovative formulation of several probiotic foods, mainly due to the inherent technological limitations of lactic acid bacteria (LAB), which represent the main probiotic microorganisms applied in food. Probiotic spores are resistant to several processing conditions employed in the food industry, thus presenting high feasibility of application and guaranteeing the functionality of probiotic foods; in addition, they are able to maintain such characteristics throughout the shelf life of such products stored at room temperature or under refrigeration. Scope and approach: This study presents a review of the main characteristics, technological implications, and types of consumption and health benefits of potential probiotic Bacillus spores. Key findings and conclusions: Spore-forming probiotics are available in different pharmaceutical forms (sachets, capsules, tablets, pills, liquid suspensions) and used in medical and veterinary fields, and more recently in the food industry. Health benefits of spore-forming probiotics should be intensively studied, evaluating their behavior in the GIT and other anatomical sites, determining the mechanisms of action involved and correlating them with the main beneficial effects on health. Spore-forming probiotics have great potential for applications in the food industry, since they present high resistance to the technological processes and physiological mechanisms of the body, which would be lethal to LAB.
... Other studies have established links between several Clostridium species and a good diet. 31,32 This strain-level phylogeny effect reflects the phylogenetic tree structure of this single species (Fig. 5A); that is, the fruit-eating phenotype is associated with a particular locations on the strain-level phylogenetic tree of this bacterial species. Therefore, even within the same species, different strains have different phenotype relationships. ...
Despite a growing interest in the gut microbiome of non-industrialized regions of the world, data linking microbiome features from such settings to diverse phenotypes remains uncommon. Here, using metagenomic data from a community-based cohort of 1,187 people from isolated villages in the Mesoamerican highlands of Western Honduras, we report 7,117 statistically robust associations spanning 788 gut microbial species (including both known and unknown taxa) and 126 phenotypes (including physical and mental health, medication use, diet, animal exposure, and social and economic measures). We report 394 new associations with mental health phenotypes alone, as well as 3,004 associations with diverse socioeconomic phenotypes. Distinctly, we also found 1,210 associations with microbiome metabolic pathways. We also report 302 significant associations after including strain-level phylogenies from 666 microbial species. Including the strain-phylogenetic information changes the overall relationship between gut microbiome and these phenotypes, and strain-level phylogenetic information enhances the observed relationship between microbiome and phenotypes as a whole. Our findings suggest new roles that gut microbiome surveillance can play in understanding broad features of individual and public health.
... Most commercial probiotic strains are inherently thermolabile and therefore must be protected from overheating. Th e exception is representatives of the genus Bacillus, which are resistant to negative environmental infl uences [36]. ...
Numerous studies in recent years have shown that the gut microbiome plays an important role in maintaining various physiological processes in the body, including digestion, metabolism, immune system function, defense against pathogens, biosynthesis of unique metabolites, elimination of toxins, and regulation of the function of the gut-brain axis. The gut microbiota is influenced by the way of birth, child’s feeding, genetic background, and lifestyle, including diet, exercises, medication, stress, and general host’s health. Intestinal microbial populations can vary significantly from person to person, including healthy individuals. Unfavorable changes in the microbial composition and in its functions are characteristic of dysbiosis and indicate pathological disorders in the body. The introduction of pro-, pre-, synbiotics and their other derivatives into the body, as well as transplantation of fecal microbiota, can restore the disturbed microbiota of the gastrointestinal tract (GIT). There is now a growing interest in functional innovative foods as ideal carriers for probiotics. However, many commercial probiotic products are ineffective because the beneficial bacteria they contain do not survive food processing, storage, and passage through the upper GIT. Th erefore, modern effective strategies are needed to improve the stability of probiotic microorganisms. One of the such strategies is a modern microencapsulation method. Using this technology in the manufacture of functional foods allows maintaining the stability of probiotic microorganisms during storage, protects them from the aggressive conditions of the GIT, and promotes their colonization on the mucous membrane of the large intestine. To achieve better protection and controlled release of probiotics, alginate microgels are most widely used as microcapsule shells.
... Probiotics are generally defined as 'live microorganisms which when administered in adequate amounts confer a health benefit on the host (Hill et al., 2014) having potential to support and maintain health in the population. Different Bacillus species are regarded as effective probiotics for human health (Anaya-Loyola et al., 2019;Bader et al., 2012;Lefevre et al., 2017;Maher, 2019;Suva et al., 2016) and have been safely used to produce alkaline-fermented and soyabased products for centuries (Inatsu et al., 2006;Ray et al., 2000;Wang and Fung, 1996). Moreover, Bacillus species have been found in the gut of healthy humans, indicating a potential role in the normal gut microbiota (Barbosa et al., 2005;Hoyles et al., 2012). ...
Previous studies using ileostomy samples from study participants demonstrated that the spore-forming probiotic Bacillus subtilis DE111® can germinate in the small intestine as early as 4 hours after ingestion. Metabolomics, proteomics and sequencing technologies, enabled further analysis of these samples for the presence of hypoglycaemic, hypolipidemic, antioxidant, anti-inflammatory and antihypertensive molecules. In the DE111 treatment group, the polyphenols trigonelline and 2,5-dihydroxybenzoic acid, orotic acid, the non-essential amino acid cystine and the lipokine 12,13-diHome were increased. DE111 also reduced acetylcholine levels in the ileostomy samples, and increased the expression of leucocyte recruiting proteins, antimicrobial peptides and intestinal alkaline phosphatases of the brush border in the small intestine. The combination of B. subtilis DE111 and the diet administered during the study increased the expression of the proteins phosphodiesterase ENPP7, ceramidase ASAH2 and the adipokine Zn-alpha-2-glycoprotein that are involved in fatty acid and lipid metabolism. Acute B. subtilis DE111 ingestion had limited detectable effect on the microbiome, with the main change being its increased presence. These findings support previous data suggesting a beneficial role of DE111 in digestion, metabolism, and immune health that appears to begin within hours of consumption.
... Genomic determinants of sporulation in B. subtilis have been investigated in this study. As bacterial spores offer advantages over non-spore forming bacteria, as spores have a higher survival rate during the acidic stomach passage and better stability during the processing and storage of the food products (Bader et al., 2012). Studies have reported a minimal set of 60 genes that are essential for sporulation in this species involved in the pre-septation (Stage 0), post-septation (Stage II), post-engulfment (Stage III-VI), spore coat formation and germination (Galperin et al., 2012). ...
Bacillus subtilis DE111® is a safe, well-tolerated commercially available spore-forming probiotic that has been clinically shown to support a healthy gut microbiome, and to promote digestive and immune health in both adults and children. Recently it was shown that this spore-forming probiotic was capable of germinating in the gastrointestinal tract as early as 3 h after ingestion. However, a better understanding of the mechanisms involved in the efficacy of DE111® is required. Therefore, the present investigation was undertaken to elucidate the functional properties of DE111® through employing a combination of in vitro functional assays and genome analysis. DE111® genome mining revealed the presence of several genes encoding acid and stress tolerance mechanisms in addition to adhesion proteins required to survive and colonize harsh gastrointestinal environment including multi subunit ATPases, arginine deiminase (ADI) pathway genes (argBDR), stress (GroES/GroEL and DnaK/DnaJ) and extracellular polymeric substances (EPS) biosynthesis genes (pgsBCA). DE111® harbors several genes encoding enzymes involved in the metabolism of dietary molecules (protease, lipases, and carbohyrolases), antioxidant activity and genes associated with the synthesis of several B-vitamins (thiamine, riboflavin, pyridoxin, biotin, and folate), vitamin K2 (menaquinone) and seven amino acids including five essential amino acids (threonine, tryptophan, methionine, leucine, and lysine). Furthermore, a combined in silico analysis of bacteriocin producing genes with in vitro analysis highlighted a broad antagonistic activity of DE111® toward numerous urinary tract, intestinal, and skin pathogens. Enzymatic activities included proteases, peptidases, esterase’s, and carbohydrate metabolism coupled with metabolomic analysis of DE111® fermented ultra-high temperature milk, revealed a high release of amino acids and beneficial short chain fatty acids (SCFAs). Together, this study demonstrates the genetic and phenotypic ability of DE111® for surviving harsh gastric transit and conferring health benefits to the host, in particular its efficacy in the metabolism of dietary molecules, and its potential to generate beneficial SCFAs, casein-derived bioactive peptides, as well as its high antioxidant and antimicrobial potential. Thus, supporting the use of DE111® as a nutrient supplement and its pottential use in the preparation of functional foods.
... Also, the massive proliferation of beneficial bacteria can competitively adhere to intestinal mucosal cells and reduce the chances of pathogenic bacteria colonization (Valeriano et al., 2017). With the use of probiotic FFs, many studies have demonstrated the promotion effect of B. subtilis on animal growth performance (Bader et al., 2012;Liu et al., 2012;Chen and Yu, 2020), and even more, studies have demonstrated the improvement of growth performance under heat stress and immune stress (Musa et al., 2019;Sokale et al., 2019;Abdelqader et al., 2020). In line with this result, our study demonstrated that the addition of B. subtilis was effective in increasing ADG, ADFI and decreasing F/G, which was related to B. subtilis metabolites such as extracellular digestive enzymes and antifungal proteins (Kim et al., 2004;Sahu et al., 2008). ...
Fermented yellow wine lees (FYWL) are widely used to increase feed utilization and improve pig performance. Based on the preparation of co-FYWL using Bacillus subtilis and Enterococcus faecalis , the purpose of this study was to investigate the effects of co-FYWL on growth performance, gut microbiota, meat quality, and immune status of finishing pigs. 75 pigs were randomized to 3 treatments (5 replicates/treatment), basal diet (Control), a basal diet supplemented with 4%FYWL, and a basal diet supplemented with 8%FYWL, for 50 days each. Results showed that the 8% FYWL group significantly reduced the F/G and increased the average daily weight gain of pigs compared to the control group. In addition, 8% FYWL improved the richness of Lactobacillus and B. subtilis in the gut, which correlated with growth performance, serum immune parameters, and meat quality. Furthermore, acetate and butyrate in the feces were improved in the FYWL group. Simultaneously, FYWL improved the volatile flavor substances of meat, increased the content of flavor amino acids, and played a positive role in the palatability of meat. In addition, FYWL increased serum IgA, IgM, IL-4 and IL-10 levels. Overall, the growth performance, the gut microbiota associated with fiber degradation, meat quality, and immune status were improved in the 8% FYWL group.
... The predominant probiotic species on the market are strains of Lactobacillus, Bifidobacterium, and Saccharomyces. However, there is increasing interest in the use of a number of different Bacillus species as safe and effective probiotics for humans (Bader et al., 2012;Suva et al., 2016;Lefevre et al., 2017;Anaya-Loyola et al., 2019;Maher, 2019). In order to be efficacious, probiotics need to reach their target location and remain viable. ...
This E-book aims to collect the latest research on the authenticity evaluation of probiotic foods and dietary supplements. It covers a total of six articles, including three original researches, two methods, and one review, with a focus on the legislation, assessment, development, and application of chemical, molecular, and omics methods to evaluate the authenticity of probiotic foods and supplements.
... The beneficial effects of Se and probiotic bacteria on growth performance of chickens have been well-documented (Yang et al., 2017;Zhang et al., 2012). B. subtilis, a probiotic Bacillus strain, has a greater ability to survive in harsh conditions (low moisture, high pelleting temperature, and less nutrients) than Lactobacillus, which has been more widely developed as a biomedical additive (Bader et al., 2012). The compound additive (SNEBS) that containing SeNPs and Bacillus subtilis would combine considerably favourable impacts of the two elements on the growth performance of lambs. ...
... Some of the benefits in spore as a probiotic candidate are surviving in acid pH at 2-2.5 (Ananthanarayanan and Dubhashi 2016), heat stability, and storage in dry form (Cutting 2011). From these benefits, bacteria in the form of spores show good stability when applied as probiotics (Bader et al. 2012;Lakshmi et al. 2017). ...
Mahariawan IMD, Kusuma WE, Yuniarti A, Beltran MAG, Hariati AM. 2021. Application of wheat flour (Triticum aestivum) on spore density and sporulation efficiency of Bacillus megaterium isolated from Litopenaeus vannamei gastrointestinal tract. Biodiversitas 22: 3709-3715. Bacillus megaterium is frequently used in fish farming, such as white shrimp (Litopenaeus vannamei) pond, which can produce spores with high stability in its implementation. Currently, spore production still requires the availability of high-cost carbon sources. The objective of this research was to evaluate the effect of different wheat flour doses on spore density and sporulation efficiency of B. megaterium BM1. In flasks, 50 mL of each test medium was treated with different doses of wheat (10, 20, 30 and 40 g. L-1, respectively) and glucose was used as a control. Each treatment was inoculated with B. megaterium BM1 (2.6 x 108 CFU. mL-1) and incubated in a shaker incubator (120 rpm) at 37 °C for 120 hours. The results showed that the highest vegetative cell concentration (17 x 108 CFU. mL-1), growth rate (0.8 hour-1) and spore (14.7 x 108 spores. mL-1) were found in the wheat flour dose of 30 g. L-1. Furthermore, the highest sporulation efficiency was achieved at 20 g. L-1 of wheat (91.30%) and germination should be done at a dose less than 40 g. L-1. The size of the spores was 1.35-1.39 µm. Thus, 30 g. L-1 of wheat flour is a potential dose to produce spore for probiotic candidates.
... The predominant probiotic species on the market are strains of Lactobacillus, Bifidobacterium, and Saccharomyces. However, there is increasing interest in the use of a number of different Bacillus species as safe and effective probiotics for humans (Bader et al., 2012;Suva et al., 2016;Lefevre et al., 2017;Anaya-Loyola et al., 2019;Maher, 2019). In order to be efficacious, probiotics need to reach their target location and remain viable. ...
Spore-based probiotics offer important advantages over other probiotics as they can survive the harsh gastric conditions of the stomach and bile salts in the small intestine, ultimately germinating in the digestive tract. A novel clinical trial in 11 ileostomy participants was conducted to directly investigate the presence and germination of the probiotic strain Bacillus subtilis DE111® in the small intestine. Three hours following ingestion of DE111®, B. subtilis spores (6.4 × 104 ± 1.3 × 105 CFU/g effluent dry weight) and vegetative cells (4.7 × 104 ± 1.1 × 105 CFU/g effluent dry weight) began to appear in the ileum effluent. Six hours after ingestion, spore concentration increased to 9.7 × 107 ± 8.1 × 107 CFU/g and remained constant to the final time point of 8 h. Vegetative cells reached a concentration of 7.3 × 107 ± 1.4 × 108 CFU/g at 7 h following ingestion. These results reveal orally ingested B. subtilis DE111® spores are able to remain viable during transit through the stomach and germinate in the small intestine of humans within 3 h of ingestion.
... Because of the low efficiency of the licensed human adjuvants, particularly in immunocompromised people, there is a great need to develop new potent and cost-effective adjuvants, which can induce a strong and long-lasting immune response (1). Spores of Bacillus subtilis, a Gram-positive bacterium, are extensively used as a probiotic in the food industry (4)(5)(6). Because of the safety of this bacterium and its potential to stimulate immune responses, it has been largely investigated for its adjuvanticity (7)(8)(9). ...
Background:
Bacterial spores are among the most efficient vaccine delivery vehicles. Because of their safety and efficacy, Bacillus subtilis spores are increasingly used in this regard. The negatively charged surfaces of the spores allow antigens to be adsorbed onto these structures. In this study, a candidate vaccine against Salmonella enterica serovar Typhi was adsorbed onto B. subtilis spores and the immunogenicity of the formulation was investigated in BALB/c mice.
Methods:
This work was performed during 2018-2019 in Islamic Azad University of Lahijan. FliC protein was recombinantly expressed in E. coli BL21 (DE3) cells and purified by affinity chromatography. On the other hand, B. subtilis strain PY79 (ATCC1609) was cultured in DSM medium and after the sporulation, FliC protein was adsorbed onto the spores in three different pH values (4, 7 and 10) and the adsorption was verified using dot-blot assay. FliC-adsorbed spores were then administered to BALB/c mice through the subcutaneous route. Mice immunization was evaluated by serum IgG assessment and challenge study.
Results:
FliC protein was successfully expressed and purified. Sporulation was controlled by phase-contrast microscopy. Serum IgG assay showed significant stimulation of the mice's humoral immune system. Immunized mice were able to resist bacterial infection.
Conclusion:
The results showed the efficiency of spores as natural adjuvants for the stimulation of mice immune system. The formulation can be exploited for the delivery of recombinant vaccines against bacterial pathogens.
... With the growing interest in probiotics and their application in animal research, more and mounting supports the concept that dietary B. subtilis or B. licheniformis could promote the growth performance of chickens (Bader and Albin et al., 2012;Liu et al.,2012;Chen and Yu, 2020). Consistent with these studies, our results demonstrated that supplementation of broiler diets with B. subtilis or B. licheniformis improved growth performance by increasing the BW, ADG, ADFI, and reducing the F:G ratio, during the starter feeding phase. ...
This study investigated the effects of dietary supplementation with Bacillus subtilis (B. subtilis) or Bacillus licheniformis (B. licheniformis) on growth performance, immunity, antioxidant capacity, short chain fatty acid (SCFA) production, and the cecal microflora in broiler chickens. In total, 360 male, 1-day-old Cobb 500 birds were randomly divided into 3 groups: the control group was fed a basal diet; the B. subtilis group was fed a basal diet supplemented with 1.5 × 10⁹ CFU/kg B. subtilis; the B. licheniformis group was fed a basal diet supplemented with 1.5 × 10⁹ CFU/kg B. licheniformis. Results showed that chickens supplemented with either B. subtilis or B. licheniformis had comparatively higher (P < 0.05) body weight and average daily gain, whereas no difference (P > 0.05) was observed in feed efficiency. Concentrations of serum IgA, IgY and IgM, as well as anti-inflammatory IL-10 were significantly increased (P < 0.05), and pro-inflammatory IL-1β and IL-6 were significantly decreased (P < 0.05) by B. subtilis or B. licheniformis supplementation. Moreover, chickens fed with diets supplemented by either B. subtilis or B. licheniformis had greater antioxidant capacity, indicated by the notable increases (P < 0.05) in glutathione peroxidase, superoxide dismutase, and catalase, along with decrease (P < 0.05) in malondialdehyde. Compared to the control group, levels of SCFA, excluding acetic and propionic acid, in cecal content had improved (P < 0.05) by adding B. licheniformis, and significant increase (P < 0.05) in acetic and butyric acid was observed with B. subtilis supplementation. Microbial analysis showed that both B. subtilis or B. licheniformis supplementation could increase butyrate-producing bacteria such as Alistipes and Butyricicoccus, and decrease pathogenic bacteria such as the Synergistetes and Gammaproteobacteria. In summary, dietary supplemented with B. subtilis or B. licheniformis improved growth performance, immune status, and antioxidant capacity, increased SCFA production, and modulated cecal microbiota in chickens. Moreover, B. licheniformis was more effective than B. subtilis with the same supplemental amount.
... offer high tolerance towards acid, dehydration, γ-ray and ultraviolet radiation; they are stable during heat processing and low-temperature storage [11][12][13][14]. Several Bacillus strains have been screened for their potential probiotic functionalities in animal husbandry, bionematicides and antibiotic alternatives [15,16]. Additionally, they have also been verified to possess pathogen exclusion, anti-oxidant, immuno-modulatory and food fermentation abilities [17][18][19][20]. ...
Foodborne diseases have been witnessing a constant rising trend worldwide, mainly caused by pathogenic microorganisms, such as Bacillus spp., posing a direct threat to public health. The purpose of this study was to evaluate the biological risk of foodborne and probiotic Bacillus spp. in Beijing markets. A total of 55 Bacillus isolates, including 29 B. cereus, 9 B. licheniformis and 7 B. subtilis, mostly found in dairy products (32.7%), were recovered from 106 samples and identified by matrix-assisted laser desorption/ionization mass spectrometry and polymerase chain reaction methods. The susceptibility towards 16 antibiotics was determined using a broth microdilution method. Bacillus showed a high level of resistance to florfenicol (100%), lincomycin (100%), tiamulin (78.2%) and ampicillin (67.3%), while they were all susceptible or intermediate to vancomycin and rifampin. Additionally, we obtained the whole genome of 19 Bacillus strains using high-throughput sequencing, and the rates of resistance genes van, fosB, erm and tet were 57.9%, 57.9%, 21.1% and 26.3%, respectively. Moreover, 100%, 9.1%, 45.5% and 100% of these isolates carried virulence genes nhe, hbl, cytK and entFM, respectively. Lastly, 60% Bacillus strains were positive in hemolysis tests, and 3 B. licheniformis strains displayed an inhibitory activity on the growth of S. aureus ATCC 29213 using agar overlay technique. Our study outlines the characteristics of foodborne Bacillus spp. and provides information for the monitoring of food safety.
... Actinomycetes can be isolated from different environmental sources including soil, water, decaying plants, and animals 11,12 . Unlike many bacterial communities including Bacillus and Clostridium which produce endospore as a dormant structure, in actinomycetes, mycolic acid and a peptidoglycan layer form a non-productive and tough structure to survive under harsh conditions for a long period of time [29][30][31][32] . ...
Recent advancements in DNA-based approaches have led to the identification of uncommon and rare bacterial pathogens. In this study, by utilizing a DNA-based approach, a total of 1043 clinical specimens were processed for the identification of actinobacteria targeting the 16S rRNA and gyrB genes. Drug susceptibility testing was also conducted using micro-broth dilution and PCR. Two isolates of Nocardia flavorosea and Rhodococcus erythropolis were reported for the first time in Iran. Also, Nocardiopsis dassonvillei, Streptomyces olivaceus, and Streptomyces griseus were reported for the first time in Asia. Infections caused by Nocardia caishijiensis and Prauserella muralis have also been reported in this study. The first Asian case of pulmonary infection caused by Nocardia ignorata and the first global case of brain abscess caused by Nocardia ninae and Nocardia neocaledoniensis have been reported in this study. Overall 30 isolates belonging to 6 genera (Nocardia, Streptomyces, Rodoccoccus, Nocardiopsis, Rothia, and
Prauserella) were detected in 30 patients. All 30 isolates were susceptible to amikacin and linezolid. Three isolates including Nocardia otitidiscaviarum (n=2) and Nocardia flavorosea (n=1) were resistant to trimethoprim-sulfamethoxazole which were the first trimethoprim-sulfamethoxazole resistant clinical actinomycetes in Iran. Isolation of rare species of actinomycetes particularly Nocardia spp. requires urgent action before they spread clinically particularly among immunocompromised patients.
... Bacillus strains with claimed probiotic properties (PB) has shown greater resistance to harsh food processing as well as gastrointestinal conditions compared to probiotic strains of Lactobacillus and Bifidobacterium, primarily because their spore-forming ability (Nithya & Halami, 2013;Soares, Almada, et al., 2019, Cao et al., 2020. The endospores of the genus Bacillus show high stability during the processing and storage of the food products (Bader, Albin, & Stahl, 2012). Further, germination of the spores does not occur, thus the quality of the product is not affected because of their inactive metabolism (Fares et al., 2015). ...
This study assessed the impact of eight unit operations [slow pasteurization, high-temperature short time (HTST) pasteurization, cooking, baking, drying, fermentation, supercritical carbon dioxide (CO2), irradiation and extrusion] in different food matrices (milk, orange juice, meatballs, bread, crystallized pineapple, yogurt, orange juice, ground black pepper, snacks, and spaghetti) on the resistance of eight (Bacillus flexus Hk1 Bacillus subtilis Bn1, Bacillus licheniformis Me1, Bacillus mojavensis KJS3, Bacillus subtilis PXN21, Bacillus subtilis PB6, Bacillus coagulans MTCC 5856 and Bacillus coagulans GBI-30, 6086) Bacillus strains with claimed probiotic properties (PB). The number of decimal reductions (γ) caused by the unit operations varied (p < 0.05) amongst the PB. Most of the unit operations caused ≤ 2 γ of PB in the food matrices evaluated. Irradiation caused up to 4.9 γ (p < 0.05) amongst the PB tested. B. subtilis Bn1, B. mojavensis KJS3, B. licheniformis Me1, and B. coagulans GBI-30 showed higher resistance to most of the tested unit operations. These results indicate that the choice of PB for application in foods should also be based on their resistance to unit operations employed during processing. Finally, the high resistance of PB to the unit operations tested comprise valuable data for the development and diversification of probiotic foods with sporeforming strains with claimed probiotic properties.
... Spores of Bacillus cereus cause food poisoning and present a challenge to the food industry [2,3], while spores of B. anthracis, which causes anthrax, are a concern because of their potential use as agents of bioterrorism and biowarfare [4]. In contrast, the durability of spores has seen their development for use as probiotics in the food industry [5,6]. Meanwhile their temperature resistance and their ability capacity to resist dessication is leading to the development of spores as delivery systems for vaccines [7]. ...
Bacillus subtilis spore coat is a bacterial proteinaceous structure with amazing characteristics of self-organization, unique resiliency, toughness and flexibility in the same time. The spore coat represents a complex multilayered protein structure which is composed of over 80 coat proteins. Some of these proteins form two dimensional crystal structures who’s low resolution ternary structure as was determined by electron microscopy. However, there are no 3D structure of these proteins known, due to a problem of preparing 3D crystals which could be analyzed by synchrotron X-ray sources. In the present study, Grazing-Incidence Wide-Angle X-ray Scattering (GIWAXS) was applied to investigate a diffraction pattern of CotY 2D crystals formed on Langmuir monolayer films. We observed two distinct diffraction rings and their position corresponds to a structure with the lattice spacing of 10.6 Å and 5.0 Å, respectively. Obtaining diffractions of 2D crystals pave the way to determination of 3D structure of coat proteins by using strong X-ray sources.
Bacillus sp. THPS1 is a novel strain isolated from a high-temperature hot spring in Thailand, exhibiting distinctive genomic features that enable adaptation to an extreme environment. This study aimed to characterize the genomic and functional attributes of Bacillus sp. THPS1 to understand its adaptation strategies and evaluate its potential for biotechnological applications. The draft genome is 5.38 Mbp with a GC content of 35.67%, encoding 5606 genes, including those linked to stress response and sporulation, which are essential for survival in high-temperature conditions. Phylogenetic analysis and average nucleotide identity (ANI) values confirmed its classification as a distinct species within the Bacillus genus. Pangenome analysis involving 19 others closely related thermophilic Bacillus species identified 1888 singleton genes associated with heat resistance, sporulation, and specialized metabolism, suggesting adaptation to nutrient-deficient, high-temperature environments. Genomic analysis revealed 12 biosynthetic gene clusters (BGCs), including those for polyketides and non-ribosomal peptides, highlighting its potential for synthesizing secondary metabolites that may facilitate its adaptation. Additionally, the presence of three Siphoviridae phage regions and 96 mobile genetic elements (MGEs) suggests significant genomic plasticity, whereas the existence of five CRISPR arrays implies an advanced defense mechanism against phage infections, contributing to genomic stability. The distinctive genomic features and functional capacities of Bacillus sp. THPS1 make it a promising candidate for biotechnological applications, particularly in the production of heat-stable enzymes and the development of resilient bioformulations.
Eman M. Ragab, Sanad T. Atallah, Elrefaey Aboelftouh, Eman M. El-Ktany* Original Research Assessment of the Impact of Bacillus Probiotics, Coconut Oil and Nigella sativa Oil on Productive Performance, Economic Indicators and Carcass Characteristics of cobb 500 Broiler Chickens This experiment was conducted to evaluate the impact of dietary inclusion of probiotic mixture (B. subtilis and B. licheniformis), Coconut oil (CO) and Nigella sativa oil (NSO) or Black cumin oil (BCO) on productive performance, economic efficiency indicators and carcass characteristics of broiler chicks. For this purpose, 140 day-old broiler chicks (Cobb500) were weighed individually and assigned randomly into four treatments (T) 35 chicks in each, divided into five replicates of 7 chicks in each. (T1) control fed a basal diet, (T2) fed a basal diet + Coconut oil (10 ml/kg feed); (T3) fed a basal diet + Nigella sativa oil (1ml/kg feed) and (T4) fed a basal diet + Bacillus probiotics (1g/kg of diet). Results clarified that CO, NSO and probiotic mixture groups improved significantly (P>0.05) the productive parameters (final body weight (FBW), body weight gain (BWG) and feed conversion ratio (FCR)) compared to control group. CO-based diet group recorded the highest BW and BWG, while probiotics mixture group was superior in feed intake value. The lowest FCR value was recorded in CO and NSO-based diet groups. The best economic efficiency measures were scored in CO followed by NSO then Bacillus probiotics. Also, there are significant differences (P<0.05) among dietary treatments on relative weight of carcass, inner organs and immune organs. Diet containing NSO acquired the highest dressing percent (73.50%), while the highest relative weight of (heart, proventriculus, thymus, spleen %) was registered in broilers fed probiotics mixture diet. In conclusion CO, NSO, probiotics mixture-based diet has a beneficial influence on productive performance and economic indicators, thus highly recommended utilizing those supplements as natural feed additives.
Probiotics with diverse and crucial properties and functions have attracted broad interest from many researchers, who adopt intestinal delivery of probiotics to modulate the gut microbiota. However, the major problems faced for the therapeutic applications of probiotics are the viability and colonization of probiotics during their processing, oral intake, and subsequent delivery to the gut. The challenges of simple oral delivery (stability, controllability, targeting, etc.) have greatly limited the use of probiotics in clinical therapies. Nanotechnology can endow the probiotics to be delivered to the intestine with improved survival rate and increased resistance to the adverse environment. Additionally, the progress in synthetic biology has created new opportunities for efficiently and purposefully designing and manipulating the probiotics. In this article, a brief overview of the types of probiotics for intestinal delivery, the current progress of different probiotic encapsulation strategies, including the chemical, physical, and genetic strategies and their combinations, and the emerging single‐cell encapsulation strategies using nanocoating methods, is presented. The action mechanisms of probiotics that are responsible for eliciting beneficial effects are also briefly discussed. Finally, the therapeutic applications of engineered probiotics are discussed, and the future trends toward developing engineered probiotics with advanced features and improved health benefits are proposed.
Aeriscardovia aeriphila, also known as Bifidobacterium aerophilum, was first isolated from the caecal contents of pigs and the faeces of cotton-top tamarin. Bifidobacterium species play important roles in preventing intestinal infections, decreasing cholesterol levels, and stimulating the immune system. In this study, we isolated a strain of bacteria from the duodenal contents of broiler chickens, which was identified as A. aeriphila, and then evaluated the effects of A. aeriphila on growth performance, antioxidant functions, immune functions, and gut microbiota in commercial broiler chickens. Chickens were orally gavaged with A. aeriphila (1×109 CFU/mL) for 21 d. The results showed that A. aeriphila treatment significantly increased the average daily gain and reduced the feed conversion ratio (P<0.001). The levels of serum growth hormone (GH) and insulin-like growth factor 1 (IGF-1) were significantly increased following A. aeriphila treatment (P<0.05). Blood urea nitrogen and aspartate aminotransferase levels were decreased, whereas glucose and creatinine levels increased as a result of A. aeriphila treatment. Furthermore, the levels of serum antioxidant enzymes, including catalase (P<0.01), superoxide dismutase (P<0.001), and glutathione peroxidase (P<0.05), and total antioxidant capacity (P<0.05) were enhanced following A. aeriphila treatment. A. aeriphila treatment significantly increased the levels of serum immunoglobulin A (IgA) (P<0.05), IgG (P<0.01), IgM (P<0.05), interleukin-1 (IL-1) (P<0.05), IL-4 (P<0.05), and IL-10 (P<0.05). The broiler chickens in the A. aeriphila group had higher secretory IgA (SIgA) levels in the duodenum (P<0.01), jejunum (P<0.001), and cecum (P<0.001) than those in the control group. The messenger RNA (mRNA) relative expression levels of IL-10 (P<0.05) and IL-4 (P<0.001) in the intestinal mucosa of chickens were increased, while nuclear factor-κB (NF-κB) (P<0.001) expression was decreased in the A. aeriphila group compared to the control group. Phylum-level analysis revealed Firmicutes as the main phylum, followed by Bacteroidetes, in both groups. The data also found that Phascolarctobacterium and Barnesiella were increased in A. aeriphila-treated group. In conclusion, oral administration of A. aeriphila could improve the growth performance, serum antioxidant capacity, immune modulation, and gut health of broilers. Our findings may provide important information for the application of A. aeriphila in poultry production.
Metabolic endotoxemia (ME) is a condition that affects many people and plays a role in the progression of various chronic diseases. ME occurs when there is a chronic, high level of bacterial endotoxin in the blood. This leads to chronic inflammation via the binding of endotoxin to Toll-like receptor 4 which results in the release of pro-inflammatory cytokines. ME is associated with high-fat diet, obesity, chronic alcohol consumption, smoking, periodontal disease, diet-induced low microbial diversity, and aging. Chronic diseases and conditions for which there is evidence that ME plays a role include atherosclerosis, diabetes, and obesity. Promising interventions for ME include lifestyle changes, interventions to increase secretory IgA and mucin production, and interventions to improve gut microbiome health. Considering the correlation between microbiota, nutrition, gastrointestinal (GI) permeability, and disease risk, we and others have speculated that these changes present treatment targets for intervention with probiotics. Spore-based Bacillus probiotics hold considerable promise for mitigating ME. This chapter details what ME is and its causes and pathophysiology, associated diseases, and interventions.Keywords
Bacillus
InflammationLipopolysaccharideMetabolic endotoxemiaMicrobiomePathophysiologyProbiotics
Spore-forming bacteria of the Bacillus genus have demonstrated potential as probiotics for human use. Bacillus clausii have been recognized as efficacious and safe agents for preventing and treating diarrhea in children and adults, with pronounced immunomodulatory properties during several in vitro and clinical studies. Herein, we characterize the novel strain of B. clausii CSI08 (Munispore®) for probiotic attributes including resistance to gastric acid and bile salts, the ability to suppress the growth of human pathogens, the capacity to assimilate wide range of carbohydrates and to produce potentially beneficial enzymes. Both spores and vegetative cells of this strain were able to adhere to a mucous-producing intestinal cell line and to attenuate the LPS- and Poly I:C-triggered pro-inflammatory cytokine gene expression in HT-29 intestinal cell line. Vegetative cells of B. clausii CSI08 were also able to elicit a robust immune response in U937-derived macrophages. Furthermore, B. clausii CSI08 demonstrated cytoprotective effects in in vitro cell culture and in vivo C. elegans models of oxidative stress. Taken together, these beneficial properties provide strong evidence for B. clausii CSI08 as a promising potential probiotic.
This research aimed to assess the impact of probiotic supplementation in the broiler diet on growth performance, nutrient utilization, noxious gas emissions, excreta micromiota and meat quality. One thousand six hundred and twenty male Ross 380 broilers (one‐day‐old, body weight, 42 ± 0.5 g and 5‐week trial) were arbitrarily chosen and assigned to three nutritive treatments (basal diet and basal diet included with 0.1%, and 0.2% probiotic mixture [Bacillus subtilis 7.0 × 107 cfu/g, Bacillus licheniformis 4.1 × 107 cfu/g]) with 30 duplicates (18 birds each). Probiotic inclusion linearly increased (p < 0.05) broiler body weight gain (BWG) during Phases 1, 2 and the overall period and decreased (p < 0.05) feed conversion ratio (FCR) linearly on Phase 2 and the overall period. However, feed intake (FI) and mortality rate remained unaffected (p > 0.05). Though nutrient digestibility of nitrogen (N) tendency to increase (p < 0.05), dry matter (DM) and energy (E) did not influence (p > 0.05). Inclusion of a probiotic supplement linearly increased (p < 0.05) Lactobacillus and reduced Salmonella (p < 0.05) counts in broilers. Moreover, broilers fed a diet supplement with probiotic addition linearly decreased (p < 0.05) NH3, H2S, C2O and acetic acid emissions. The graded level of probiotic addition linearly reduced (p < 0.05) cooking loss and the tendency to decrease (p < 0.05) weight of bursa of Fabricius, but had no effect (p > 0.05) on other meat quality measures. These findings indicated that increasing the level of probiotics in feed could improve growth efficiency, nutrient absorption, microbial index, meat quality and reduce gas emissions in broilers.
Bacillus coagulans (B. coagulans) can improve and prevent functional gastrointestinal disorders. However, there has been little discussion in the literature on the difference between spores and vegetative cells for relieving...
The usage of probiotic bacteria in aquaculture is profoundly ranging from growth promotion of cultured fishes to improvement of the rearing environment. Among the different groups of probiotic bacteria, the Bacillus spp. stand out due to their capability to withstand the harsh environment in nature and in fish gut. Although the probiotic Bacillus spp. in sustainable aquaculture is widely applicable, the mechanisms of their beneficial effects are not comprehensively reviewed. This chapter aimed to update our knowledge of the mode of actions of Bacillus spp. as probiotic in the promotion of aquaculture. The probiotic Bacillus spp. secrete diverse enzymes that help fishes in the digestion of complex feedstuffs thus decrease the feed conversion ratio and increase the specific growth rate of the host fishes. They can colonize the fish gut and produce diverse antimicrobial substances like bacteriocins (antibiotics) and lytic enzymes and promote immunity in fishes to protect from pathogens. They also compete for nutrient, energy, and binding sites and thus exclude pathogens from the fish guts. The probiotic Bacillus spp. also boost the host’s genes expression associated with the growth and development of the fishes. Conversion of toxic nitrogenous wastes by Bacillus spp. detoxifies the rearing water and environment. This chapter is compendium of existing knowledge on the mechanism of beneficial actions of Bacillus spp. in aquaculture.KeywordsGreen aquacultureBiocontrol of fish diseasesQuorum quenchingGene expressionISRFish probiotics
Nowadays, the treatment of gastrointestinal disorders with probiotics is a
widely used remedy for intestinal complications in humans. This essential
sources of microorganisms was originally developed for the treatment of
health illness, based on the ability to functioning the normal pathway of
intestine moreover, the clinical effects were reported as a remarkable
advancement over commonly used treatment in GIT problems. Subsequently, it
was realized that the clinical applicability of the enhanced intestinal floral
development of used probiotics has been expanded to the therapeutic
management of other gastrointestinal disorders, including cancer, heart
failure etc.
The most well known food-based strategies to modulate the composition of the intestinal microbiota are the dietary use of prebiotics, probiotics and their combination, synbiotics. Currently established prebiotic compounds are mainly targeting the bifidobacteria population of the colon microbiota. A good illustration of the importance of high colonic bifidobacteria levels is the observation that breast milk creates an environment in the colon (because of its high amount in galacto-oligosaccharides with prebiotic activity) favouring the development of a simple flora, dominated by bifidobacteria to which various health benefits have been ascribed. Currently, high colonic bifidobacteria levels has been considered favourably at all ages and strategies to augment their presence have been demonstrated in placebo-controlled intervention studies; e.g. in toddlers to reduce sickness events, in adults to reduce the risk for developing gastrointestinal diseases and in the elderly to re-enhance their declining immune activity. The intestinal microbiota can be considered as a metabolically adaptable and rapidly renewable organ of the body. However, unbalances in its microbial community and activities are found to be implicated in disease initiation and progression, such as chronic inflammatory bowel diseases and colonic cancers. Restoration of this balance by increasing bifidobacteria levels has demonstrated to reduce disease severity of patients and to improve well-being in healtly volunteers. New emerging evidence on the difference in the composition of the colonic microbiota between obese and lean volunteers has opened new areas for pre-, pro- and synbiotic research. Additionally, as knowledge will increase about the microbial bio-conversion of polyphenolic compounds into bioactive metabolites in the colon and whether food-based strategies can augment such bioconversion into more potent compounds with anti-oxidant and/or anti-inflammatory activity new areas of research will be discovered. This paper provides an up-to-date review of the health benefits associated to the induction of high bifidobacteria levels in the colon by the use of prebiotics (inulin and oligofructose). New areas of emerging science will be discussed as well.
The present paper summarizes the consensus views of a group of 9 European clinicians and scientists on the current state of scientific knowledge on probiotics, covering those areas where there is substantial evidence for beneficial effects and those where the evidence base is poor or inconsistent. There was general agreement that probiotic effects were species and often strain specific. The experts agreed that some probiotics were effective in reducing the incidence and duration of rotavirus diarrhoea in infants, antibiotic-associated diarrhoea in adults and, for certain probiotics, Clostridium difficile infections. Some probiotics are associated with symptomatic improvements in irritable bowel syndrome and alleviation of digestive discomfort. Probiotics can reduce the frequency and severity of necrotizing enterocolitis in premature infants and have been shown to regulate intestinal immunity. Several other clinical effects of probiotics, including their role in inflammatory bowel disease, atopic dermatitis, respiratory or genito-urinary infections or H.pylori adjuvant treatment were thought promising but inconsistent.
Coevolution of mammals and their gut microbiota has profoundly affected their radiation into myriad habitats. We used shotgun sequencing of microbial community DNA and targeted sequencing of bacterial 16S ribosomal RNA genes to gain an understanding of how microbial communities adapt to extremes of diet. We sampled fecal DNA from 33 mammalian species and 18 humans who kept detailed diet records, and we found that the adaptation of the microbiota to diet is similar across different mammalian lineages. Functional repertoires of microbiome genes, such as those encoding carbohydrate-active enzymes and proteases, can be predicted from bacterial species assemblages. These results illustrate the value of characterizing vertebrate gut microbiomes to understand host evolutionary histories at a supraorganismal level.
Microbial colonization of mammals is an evolution-driven process that modulate host physiology, many of which are associated with immunity and nutrient intake. Here, we report that colonization by gut microbiota impacts mammalian brain development and subsequent adult behavior. Using measures of motor activity and anxiety-like behavior, we demonstrate that germ free (GF) mice display increased motor activity and reduced anxiety, compared with specific pathogen free (SPF) mice with a normal gut microbiota. This behavioral phenotype is associated with altered expression of genes known to be involved in second messenger pathways and synaptic long-term potentiation in brain regions implicated in motor control and anxiety-like behavior. GF mice exposed to gut microbiota early in life display similar characteristics as SPF mice, including reduced expression of PSD-95 and synaptophysin in the striatum. Hence, our results suggest that the microbial colonization process initiates signaling mechanisms that affect neuronal circuits involved in motor control and anxiety behavior.
Probiotics, live cells with different beneficiary characteristics, have been extensivelly studied and explored commercially in many different products in the world. Their benefits to human and animal health have been proven in hundreds of scientific research. Lactobacillus and Bifidobacterium are the main probiotic groups; however, there are reports on the probiotic potential of Pediococcus, Lactococcus, Bacillus and yeasts. Some of the identified probiotic strains exhibit powerful anti-inflammatory, antiallergic and other important properties. Apart from that, the consumption of dairy and non-dairy products stimulates the immunity in different ways. Various food matrices have been used with probiotics, which are briefly documented. In this review, the history of probiotics, their application in the health and food areas and new trends in probiotic products and processes are presented.
There is increasing evidence that the postprandial state is an important contributing factor to chronic disease. The role of fruit phenolic compounds to protect health and lower disease risk through their actions in mitigating fed-state metabolic and oxidative stressors is of interest and the topic of the present paper. Two main questions are posed: first, what is the role of plant foods, specifically fruits rich in complex and simple phenolic compounds in postprandial metabolic management; and second, does the evidence support consuming these fruits with meals as a practical strategy to preserve health and lower risk for disease? This review provides an overview of the postprandial literature, specifically on the effect of fruits and their inherent phenolic compounds in human subjects on postprandial lipaemia, glycaemia/insulinaemia and associated events, such as oxidative stress and inflammation. Among the identified well-controlled human trials using a postprandial paradigm, >50 % of the trials used wine or wine components and the remaining used various berries. Notwithstanding the need for more research, the collected data suggest that consuming phenolic-rich fruits increases the antioxidant capacity of the blood, and when they are consumed with high fat and carbohydrate 'pro-oxidant and pro-inflammatory' meals, they may counterbalance their negative effects. Given the content and availability of fat and carbohydrate in the Western diet, regular consumption of phenolic-rich foods, particularly in conjunction with meals, appears to be a prudent strategy to maintain oxidative balance and health.
This work was aimed at identifying strains which can degrade quorum-sensing (QS) molecules from fish gut, with properties suitable for use as probiotic in aquaculture.
A total of 200 strains were obtained from the intestine gut of Carassius auratus gibelio after enrichment in KG medium contained 500 μg l(-1) of C6-HSL as the sole source of carbon and nitrogen; one strain named QS inhibitor (QSI)-1 was identified as the genus Bacillus spp. by morphological phenotypes, and the strain also possessed an aiiA homologue gene using PCR amplification. In vitro, QSI-1 strongly interfered with violacein production by Chromobacterium violaceum. Coculture of QSI-1 with fish pathogen effectively reduced the amount of acyl-homoserine lactones (AHLs) and the extracellular proteases activity of Aeromonas hydrophila YJ-1. The oral LD50 of QSI-1 to fish was more than 10(11) CFU shown that it was avirulent to fish. Fish fed diet supplemented with QSI-1 had good survival, suggesting that QSI-1 showed protection against Aer. hydrophila infection.
The results indicate that the isolate QSI-1 might have the potential possibility to be used as a probiotic in aquaculture.
This is the first report to describe a bacterium isolated from the intestine gut of C. auratus gibelio which can degrade AHLs and has the probiotic characteristics for its use in aquaculture.
The different compartments of the gastrointestinal tract are inhabited by populations of micro-organisms. By far the most important predominant populations are in the colon where a true symbiosis with the host exists that is a key for well-being and health. For such a microbiota, 'normobiosis' characterises a composition of the gut 'ecosystem' in which micro-organisms with potential health benefits predominate in number over potentially harmful ones, in contrast to 'dysbiosis', in which one or a few potentially harmful micro-organisms are dominant, thus creating a disease-prone situation. The present document has been written by a group of both academic and industry experts (in the ILSI Europe Prebiotic Expert Group and Prebiotic Task Force, respectively). It does not aim to propose a new definition of a prebiotic nor to identify which food products are classified as prebiotic but rather to validate and expand the original idea of the prebiotic concept (that can be translated in 'prebiotic effects'), defined as: 'The selective stimulation of growth and/or activity(ies) of one or a limited number of microbial genus(era)/species in the gut microbiota that confer(s) health benefits to the host.' Thanks to the methodological and fundamental research of microbiologists, immense progress has very recently been made in our understanding of the gut microbiota. A large number of human intervention studies have been performed that have demonstrated that dietary consumption of certain food products can result in statistically significant changes in the composition of the gut microbiota in line with the prebiotic concept. Thus the prebiotic effect is now a well-established scientific fact. The more data are accumulating, the more it will be recognised that such changes in the microbiota's composition, especially increase in bifidobacteria, can be regarded as a marker of intestinal health. The review is divided in chapters that cover the major areas of nutrition research where a prebiotic effect has tentatively been investigated for potential health benefits. The prebiotic effect has been shown to associate with modulation of biomarkers and activity(ies) of the immune system. Confirming the studies in adults, it has been demonstrated that, in infant nutrition, the prebiotic effect includes a significant change of gut microbiota composition, especially an increase of faecal concentrations of bifidobacteria. This concomitantly improves stool quality (pH, SCFA, frequency and consistency), reduces the risk of gastroenteritis and infections, improves general well-being and reduces the incidence of allergic symptoms such as atopic eczema. Changes in the gut microbiota composition are classically considered as one of the many factors involved in the pathogenesis of either inflammatory bowel disease or irritable bowel syndrome. The use of particular food products with a prebiotic effect has thus been tested in clinical trials with the objective to improve the clinical activity and well-being of patients with such disorders. Promising beneficial effects have been demonstrated in some preliminary studies, including changes in gut microbiota composition (especially increase in bifidobacteria concentration). Often associated with toxic load and/or miscellaneous risk factors, colon cancer is another pathology for which a possible role of gut microbiota composition has been hypothesised. Numerous experimental studies have reported reduction in incidence of tumours and cancers after feeding specific food products with a prebiotic effect. Some of these studies (including one human trial) have also reported that, in such conditions, gut microbiota composition was modified (especially due to increased concentration of bifidobacteria). Dietary intake of particular food products with a prebiotic effect has been shown, especially in adolescents, but also tentatively in postmenopausal women, to increase Ca absorption as well as bone Ca accretion and bone mineral density. Recent data, both from experimental models and from human studies, support the beneficial effects of particular food products with prebiotic properties on energy homaeostasis, satiety regulation and body weight gain. Together, with data in obese animals and patients, these studies support the hypothesis that gut microbiota composition (especially the number of bifidobacteria) may contribute to modulate metabolic processes associated with syndrome X, especially obesity and diabetes type 2. It is plausible, even though not exclusive, that these effects are linked to the microbiota-induced changes and it is feasible to conclude that their mechanisms fit into the prebiotic effect. However, the role of such changes in these health benefits remains to be definitively proven. As a result of the research activity that followed the publication of the prebiotic concept 15 years ago, it has become clear that products that cause a selective modification in the gut microbiota's composition and/or activity(ies) and thus strengthens normobiosis could either induce beneficial physiological effects in the colon and also in extra-intestinal compartments or contribute towards reducing the risk of dysbiosis and associated intestinal and systemic pathologies.
Evidence suggests that probiotics reduce rotavirus diarrhoea duration. Although there are several probiotic strains potentially useful, daily practice is often limited by the type and number of products locally available. In general, information about combined products is scarce. In this study we compare the effect of two probiotic products in the treatment of diarrhoea in children less than 2 years of age.
A Randomized double-blind controlled clinical trial in children hospitalized for acute rotavirus diarrhoea, in the Paediatric Centre Albina Patino, Cochabamba, Bolivia.Participants were children aged 1 - 23 months, who were randomly assigned to receive one of three treatments: Oral rehydration therapy plus placebo; Oral rehydration solution plus Saccharomyces boulardii; or Oral rehydration solution plus a compound containing Lactobacillus acidophilus, Lactobacillus rhamnosus, Bifidobacterium longum and Saccharomyces boulardii. Sample size was 20 per group and the outcomes were duration of diarrhoea, of fever, of vomiting and of hospitalization.
64 cases finished the protocol. On admission, patients' characteristics were similar. Median duration of diarrhoea (p = 0.04) in children who received the single species product (58 hours) was shorter than in controls (84.5 hrs). Comparing children that received the single probiotic product and controls showed shorter duration of fever (18 vs 67 hrs) (p = 0.0042) and the mixed probiotic of vomiting (0 vs 42.5 hrs) (p = 0.041). There was no effect on duration of hospitalization (p = 0.31). When experimental groups were merged, statistical significance of changes increased (total duration of diarrhoea, fever and vomiting P = 0.025, P = 0.025 and P = 0.014, respectively).
Both products decreased the duration of diarrhoea compared to oral rehydration solution alone. This decrease was significant only for the single species product which also decreased the duration of fever. With the multiple species product there was no vomiting subsequent to the initiation of treatment. The quantity of probiotic bacteria needed for optimum treatment of gastroenteritis remains to be determined, particularly when multiple species are included in the product.Trial registration: ClinicalTrials.gov ID: NCT00981877Link: https://register.clinicaltrials.gov/prs/app/action/SelectProtocol/sid/S0002653/selectaction/View/ts/2/uid/U0000N04
Clinical trials NCT ID: NCT00981877.
To investigate the immune response of peripheral blood mononuclear cells (PBMCs) and dendritic cells (DCs) that were stimulated by probiotic preparations.
PBMCs were isolated, cultured, and stimulated with Bio-Three (a mixture of Bacillus mesentericus, Clostridium butyricum and Enterococcus faecalis; 10(5), 10(6) and 10(7) CFU/mL for 24 h). Cytokine production of (1) circulating PBMCs; (2) PBMCs stimulated by probiotic preparation; (3) monocyte-derived DCs; and (4) DC and T cell co-culture was determined by enzyme-linked immunosorbent assay. Phenotypic analysis of circulating PBMCs was also investigated by flow cytometry. Blood was obtained from individuals who consumed Bio-Three (10(9) CFU/d B. mesentericus, C. butyricum and E. faecalis) for 2 wk, or those who did not take probiotics orally.
In culture supernatants, interferon-gamma (IFN-gamma) and interleukin (IL)-10 production increased, but IL-4 and tumor necrosis factor-alpha (TNF-alpha) production by PBMCs decreased after 1 and 2 wk of probiotic treatment. Flow cytometry was also performed on day 14 and detected enhanced expression of CD11b, HLA-DR, CD4, CD45RA, CD25, CD44 and CD69 in response to Bio-Three. Furthermore, IL-10 and IL-12 were upregulated in supernatants of monocyte-derived DCs, and IFN-gamma and IL-10 were enhanced in supernatants of CD4(+) T cells co-cultured with DCs.
Bio-Three appeared to stimulate the Th1 immune response, downregulate pro-inflammatory cytokines (TNF-alpha) and upregulate anti-inflammatory cytokine (IL-10). Probiotics could be effective in activation of PBMCs and DCs.
This study was performed to evaluate anti-inflammatory and immune modulating properties of the probiotic, spore-forming bacterial strain: Bacillus coagulans: GBI-30, (PTA-6086, GanedenBC30TM). In addition, cell wall and metabolite fractions were assayed separately to address whether biological effects were due to cell wall components only, or whether secreted compounds from live bacteria had additional biological properties. The spores were heat-activated, and bacterial cultures were grown. The culture supernatant was harvested as a source of metabolites (MTB), and the bacteria were used to isolate cell wall fragments (CW). Both of these fractions were compared in a series of in vitro assays.
Both MTB and CW inhibited spontaneous and oxidative stress-induced ROS formation in human PMN cells and increased the phagocytic activity of PMN cells in response to bacteria-like carboxylated fluorospheres. Both fractions supported random PMN and f-MLP-directed PMN cell migration, indicating a support of immune surveillance and antibacterial defense mechanisms. In contrast, low doses of both fractions inhibited PMN cell migration towards the inflammatory mediators IL-8 and LTB4. The anti-inflammatory activity was strongest for CW, where the PMN migration towards IL-8 was inhibited down to dilutions of 1010.Both MTB and CW induced the expression of the CD69 activation marker on human CD3- CD56+ NK cells, and enhanced the expression of CD107a when exposed to K562 tumor cells in vitro.The fractions directly modulated cytokine production, inducing production of the Th2 cytokines IL-4, IL-6, and IL-10, and inhibiting production of IL-2.Both fractions further modulated mitogen-induced cytokine production in the following manner: Both fractions enhanced the PHA-induced production of IL-6 and reduced the PHA-induced production of TNF-alpha. Both fractions enhanced the PWM-induced production of TNF-alpha and IFN-gamma. In addition, MTB also enhanced both the PHA- and the PWM-induced expression of IL-10.
The data suggest that consumption of GanedenBC30TM may introduce both cell wall components and metabolites that modulate inflammatory processes in the gut. Both the cell wall and the supernatant possess strong immune modulating properties in vitro. The anti-inflammatory effects, combined with direct induction of IL-10, are of interest with respect to possible treatment of inflammatory bowel diseases as well as in support of a healthy immune system.
The first exposure to microorganisms at mucosal surfaces is critical for immune maturation and gut health. Facultative anaerobic bacteria are the first to colonise the infant gut, and the impact of these bacteria on intestinal epithelial cells (IEC) may be determinant for how the immune system subsequently tolerates gut bacteria.
To mirror the influence of the very first bacterial stimuli on infant IEC, we isolated IEC from mouse foetuses at gestational day 19 and from germfree neonates. IEC were stimulated with gut-derived bacteria, Gram-negative Escherichia coli Nissle and Gram-positive Lactobacillus acidophilus NCFM, and expression of genes important for immune regulation was measured together with cytokine production. E. coli Nissle and L. acidophilus NCFM strongly induced chemokines and cytokines, but with different kinetics, and only E. coli Nissle induced down-regulation of Toll-like receptor 4 and up-regulation of Toll-like receptor 2. The sensitivity to stimulation was similar before and after birth in germ-free IEC, although Toll-like receptor 2 expression was higher before birth than immediately after.
In conclusion, IEC isolated before gut colonisation occurs at birth, are highly responsive to stimulation with gut commensals, with L. acidophilus NCFM inducing a slower, but more sustained response than E. coli Nissle. E. coli may induce intestinal tolerance through very rapid up-regulation of chemokine and cytokine genes and down-regulation of Toll-like receptor 4, while regulating also responsiveness to Gram-positive bacteria.
Lactic acid-producing bacteria (LAB) probiotics demonstrate immunomodulating and anti-inflammatory effects and the ability to lessen the symptoms of arthritis in both animals and humans. This randomized, double-blind, placebo-controlled, parallel-design, clinical pilot trial was conducted to evaluate the effects of the LAB probiotic preparation, Bacillus coagulans GBI-30, 6086, on symptoms and measures of functional capacity in patients with rheumatoid arthritis (RA) in combination with pharmacological anti-arthritic medications.
Forty-five adult men and women with symptoms of RA were randomly assigned to receive Bacillus coagulans GBI-30, 6086 or placebo once a day in a double-blind fashion for 60 days in addition to their standard anti-arthritic medications. Arthritis activity was evaluated by clinical examination, the American College of Rheumatology (ACR) criteria, the Stanford Health Assessment Questionnaire Disability Index (HAQ-DI), and laboratory tests for erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP).
Subjects who received Bacillus coagulans GBI-30, 6086 experienced borderline statistically significant improvement in the Patient Pain Assessment score (P = .052) and statistically significant improvement in Pain Scale (P = .046) vs placebo. Compared with placebo, Bacillus coagulans GBI-30, 6086 treatment resulted in greater improvement in patient global assessment and self-assessed disability; reduction in CRP; as well as the ability to walk 2 miles, reach, and participate in daily activities. There were no treatment-related adverse events reported throughout this study.
Results of this pilot study suggest that adjunctive treatment with Bacillus coagulans GBI-30, 6086 LAB probiotic appeared to be a safe and effective for patients suffering from RA. Because of the low study population size, larger trials are needed to verify these results.
ACTRN12609000435280.
To assess whether the probiotic food supplements, produced and distributed on the Italian market during 2005-2006, complied with the Italian Guidelines on Prebiotics and Probiotics, 72 samples from 29 processing plants were analyzed. The survey included 41 samples from processing plants and 31 samples of the same brand from retailers collected at timed intervals (3, 8 and 13 months). A polyphasic approach based on a suitable analytical collection method (genotypic identification of total bacteria - differential presumptive enumeration - genotypic identification of viable bacteria) was adopted to identify and quantify the microorganisms labelled and recovered from the probiotic supplements examined. Most supplements analyzed (87%) did not conform to the Italian guidelines and the differences were both quantitative and qualitative (number determination, purity, types and viability of microorganisms). Even though most labelled supplements (25 samples) indicated the presence of Bifidobacterium bifidum, this organism was only detected sporadically and always as dead cells. Unexpected results were obtained during our survey due to the absence of viability of Bacillus coagulans spores in some labelled supplements. Besides this, some of these supplements also contained other spore-forming species, identified as B. cereus that are toxin producing. We have also documented a widespread use of misclassified microbial species or species with fictitious names. The main factors involved in the absence of compliance were examined and the poor quality control applied by manufacturers was emphasized.
This randomized double blind placebo controlled dual site clinical trial compared a probiotic dietary supplement to placebo regarding effects on gastrointestinal symptoms in adults with post-prandial intestinal gas-related symptoms (abdominal pain, distention, flatulence) but no gastrointestinal (GI) diagnoses to explain the symptoms.
Sixty-one adults were enrolled (age 36.5 +/- 12.6 years; height 165.1 +/- 9.2 cm; weight 75.4 +/- 17.3 kg) and randomized to either Digestive Advantage Gas Defense Formula - (GanedenBC30 Bacillus coagulans GBI-30, 6086): n = 30; or Placebo: n = 31. Study subjects were evaluated every two weeks over a four-week period using validated questionnaires and standard biochemical safety testing. Outcome criteria of interest included change from baseline in Gastrointestinal Symptom Rating Scale (GSRS) abdominal pain, abdominal distention, flatus, and the Severity of Dyspepsia Assessment (SODA) bloating and gas subscores over four weeks of product use.
Measured against the placebo, subjects in the probiotic group achieved significant improvements in GSRS abdominal pain subscore (p = 0.046) and the GSRS total score (p = 0.048), with a strong trend for improvement on the GSRS abdominal distension subscore (p = 0.061). A strong placebo effect was evident which could explain the lack of statistical significant differences between the groups for many of the efficacy variables.
In conclusion, the Bacillus coagulans-based product was effective in improving the quality of life and reducing gastrointestinal symptoms in adults with post prandial intestinal gas-related symptoms and no GI diagnoses.
ClinicalTrials.gov Identifier: NCT00881322.
The bacterial spore, the hardiest known life form, can survive in a metabolically dormant state for many years and can withstand high temperatures, radiation, and toxic chemicals. The molecular basis of spore dormancy and resistance is not understood, but the physical state of water in the different spore compartments is thought to play a key role. To characterize this water in situ, we recorded the water (2)H and (17)O spin relaxation rates in D(2)O-exchanged Bacillus subtilis spores over a wide frequency range. The data indicate high water mobility throughout the spore, comparable with binary protein-water systems at similar hydration levels. Even in the dense core, the average water rotational correlation time is only 50 ps. Spore dormancy therefore cannot be explained by glass-like quenching of molecular diffusion but may be linked to dehydration-induced conformational changes in key enzymes. The data demonstrate that most spore proteins are rotationally immobilized, which may contribute to heat resistance by preventing heat-denatured proteins from aggregating irreversibly. We also find that the water permeability of the inner membrane is at least 2 orders of magnitude lower than for model membranes, consistent with the reported high degree of lipid immobilization in this membrane and with its proposed role in spore resistance to chemicals that damage DNA. The quantitative results reported here on water mobility and transport provide important clues about the mechanism of spore dormancy and resistance, with relevance to food preservation, disease prevention, and astrobiology.
Bacillus subtilis grows in the absence of oxygen using nitrate ammonification and various fermentation processes. Lactate, acetate, and 2,3-butanediol
were identified in the growth medium as the major anaerobic fermentation products by using high-performance liquid chromatography.
Lactate formation was found to be dependent on the lctEP locus, encoding lactate dehydrogenase and a putative lactate permease. Mutation of lctE results in drastically reduced anaerobic growth independent of the presence of alternative electron acceptors, indicating
the importance of NADH reoxidation by lactate dehydrogenase for the overall anaerobic energy metabolism. Anaerobic formation
of 2,3-butanediol via acetoin involves acetolactate synthase and decarboxylase encoded by the alsSD operon. Mutation ofalsSD has no significant effect on anaerobic growth. Anaerobic acetate synthesis from acetyl coenzyme A requires phosphotransacetylase
encoded by pta. Similar to the case for lctEP, mutation of pta significantly reduces anaerobic fermentative and respiratory growth. The expression of both lctEP and alsSD is strongly induced under anaerobic conditions. Anaerobic lctEP andalsSD induction was found to be partially dependent on the gene encoding the redox regulator Fnr. The observed fnrdependence might be the result of Fnr-induced arfM(ywiD) transcription and subsequent lctEP andalsSD activation by the regulator ArfM (YwiD). The two-component regulatory system encoded by resDE is also involved in anaerobic lctEP induction. No directresDE influence on the redox regulation ofalsSD was observed. The alternative electron acceptor nitrate represses anaerobic lctEP andalsSD transcription. Nitrate repression requiresresDE- and fnr-dependent expression ofnarGHJI, encoding respiratory nitrate reductase. The genealsR, encoding a regulator potentially responding to changes of the intracellular pH and to acetate, is essential for anaerobic
lctEP and alsSD expression. In agreement with its known aerobic function, no obvious oxygen- or nitrate-dependent pta regulation was observed. A model for the regulation of the anaerobic fermentation genes in B. subtilis is proposed.
Several Bacillus strains isolated from commercial probiotic preparations were identified at the species level, and their adhesion capabilities to three different model intestinal surfaces (mucin, Matrigel and Caco-2 cells) were assessed. In general, adhesion of spores was higher than that of vegetative cells to the three matrices, and overall strain Bacillus cereus(CH) displayed the best adhesion. Different biochemical treatments revealed that surface proteins of B. cereus(CH) were involved in the adhesion properties of the strain. Surface-associated proteins from vegetative cells and spores of B. cereus(CH) were extracted and identified, and some proteins such as S-layer components, flagellin and cell-bound proteases were found to bind to mucin or fibronectin. These facts suggest that those proteins might play important roles in the interaction of this probiotic Bacillus strain within the human gastrointestinal tract.
The capability of endospores of Bacillus subtilis to withstand extreme environmental conditions is secured by several attributes. One of them, the protein shell that encases the spore and is known as the coat, provides the spore with its characteristic resistance to toxic chemicals, lytic enzymes, and predation by unicellular and multicellular eukaryotes. Despite most of the components of the spore coat having been identified, we have only a vague understanding of how such a complex structure is assembled. Using the yeast two-hybrid system, we attempted to identify direct contacts among the proteins allocated to the insoluble fraction of the spore coat: CotV, CotW, CotX, CotY, and CotZ. We also examined whether they could interact with CotE, one of the most crucial morphogenetic proteins governing outer coat formation and also present in the insoluble fraction. Out of all 21 possible interactions we tested, 4 were found to be positive. Among these interactions, we confirmed the previous observation that CotE forms homo-oligomers. In addition, we observed homotypic interactions of CotY, strong interactions between CotZ and CotY, and relatively weak, yet significant, interactions between CotV and CotW. The results of this yeast two-hybrid analysis were confirmed by size exclusion chromatography of recombinant coat proteins and a pull-down assay.
The effect of Bacillus natto on T and B lymphocytes in the chicken spleen was investigated. Bacillus natto is isolated from fermented food and is used in Japan in a preparation that enhances growth of farm animals. When chickens were fed 10(7) colony-forming units of B. natto (in spores) per gram of feed from hatching to 27 days of age there was an increase in the percentage of T and B lymphocytes in the spleens compared with non-B. natto-fed controls. The findings suggested that dietary Bacillus natto had an effect on cellular immune responses.
Probiotics have been suggested to improve gastrointestinal health in humans. To investigate the effects of Bacillus polyfermenticus SCD administration on fecal microflora and putrefactive metabolites in humans, Bacillus polyfermenticus SCD (4.00×105CFU/mg) was administrated to ten healthy subjects (5 men and 5 women, average age 24 years) three times a day for 2 weeks. Fecal samples were collected before (1st and 2nd weeks, control), during (3rd and 4th weeks), and 2 weeks after the administration. The following microbial groups were evaluated in the feces: aerobic and anaerobic bacteria, Bacillus polyfermenticus SCD, Lactobacillus, Bifidobacterium, total lactic acid bacteria, Salmonella, Clostridium, Clostridium perfringens, Eubacterium, Staphylococcus, Coliform bacteria, Pseudomonas, and Yeast. Fecal concentrations of total aerobic bacteria (p < 0.05, p < 0.01, 3rd and 4th weeks), total lactic acid bacteria (p < 0.01, 3rd, 4th and 5th weeks), and Bifidobacteria (p < 0.05, 4th and 5th weeks) were significantly increased in all subjects, compared to the control, from the 3rd week after the administration of the products. Clostridium (p < 0.01, 4th week), Clostridium perfringens (p < 0.05, p<0.01, 3rd and 4th weeks), and coliform (p < 0.01, 5th week) were significantly reduced from the 3rd week of administration. No significant changes in the fecal concentrations of Pseudomonas, Lactobacillus, Eubacterium, Staphylococcus, yeast, and total anaerobes were observed. Six weeks after the administration, the concentration of all microorganisms returned to the basal level. Bacillus polyfermenticus SCD was significantly maintained from the 3rd week to 6th week of the study. Despite the absence of a statistical significance, the putrefactive metabolites (ammonia, indole, skatole, and ρ-cresol) and the pH value tended to be lower during and after the test periods than the base line. These results show that this probiotic preparation is able to colonize the intestine, and suggest that it may be useful as a beneficial probiotic in humans.
While many food science programs offer courses in the microbiology and processing of fermented foods, no recently published texts exist that fully address the subject. Food fermentation professionals and researchers also have lacked a single book that covers the latest advances in biotechnology, bioprocessing, and microbial genetics, physiology, and taxonomy. In Microbiology and Technology of Fermented Foods, Robert Hutkins has written the first text on food fermentation microbiology in a generation. This authoritative volume also serves as a comprehensive and contemporary reference book. A brief history and evolution of microbiology and fermented foods, an overview of microorganisms involved in food fermentations, and their physiological and metabolic properties provide a foundation for the reader. How microorganisms are used to produce fermented foods and the development of a modern starter culture industry are also described. Successive chapters are devoted to the major fermented foods produced around the world with coverage including microbiological and technological features for manufacture of these foods: Cultured Dairy Products. Cheese. Meat Fermentation. Fermented Vegetables. Bread Fermentation. Beer Fermentation. Wine Fermentation. Vinegar Fermentation. Fermentation of Foods in the Orient. Examples of industrial processes, key historical events, new discoveries in microbiology, anecdotal materials, case studies, and other key information are highlighted throughout the book. Comprehensively written in a style that encourages critical thinking, Microbiology and Technology of Fermented Foods will appeal to anyone dealing in food fermentation - students, professors, researchers, and industry professionals.
AIM: To evaluate the therapeutic effects of a probiotic supplement (Clostridium butyricum, CGMCC0313) in a chemically-induced rat model of experimental colitis.
METHODS: An experimental ulcerative colitis model was established by rectal injection of oxazolone into the colon of 40 Wistar rats randomly divided into four groups. The positive control group was sacrificed 3 d after colitis onset. The remaining groups were fed daily with either 2 mL of C. butyricum (2.3 × 1011 CFU/L), 2 mL of mesalamine (100 g/L), or 1 mL of sodium butyrate (50 mmol/L) for 21 d. The animals’ body weight, behavior, and bowel movements were recorded weekly. After sacrifice, visual and microscopic observations of pathological changes of colon tissue were made, body weight and wet colon mass index were measured and recorded, and serum levels of interleukin-23 (IL-23) and TNF-α were measured using ELISA. Expression of calcitonin gene-related peptide in colon tissue was measured by RT-PCR. Finally, changes in rat intestinal microflora status were measured in all groups.
RESULTS: We found that treatment with C. butyricum lowered the serum levels of both IL-23 and tumor necrosis factor-α (TNF-α) with similar or even better efficiency than that of mesalamine or sodium butyrate. The rat intestinal flora appeared to recover more quickly in the group treated with C. butyricum than in the mesalamine and sodium butyrate groups. Finally, we found that the expression level of calcitonin gene related peptide was elevated in colon tissue in the sodium butyrate treated group but not in the C. butyricum or mesalamine treated groups, indicating a sensitization of colon following sodium butyrate treatment.
CONCLUSION: In our experimental colitis model, treatment with C. butyricum CGMCC0313, a probiotic supplement, is at least as efficient as treatment with mesalamine.
A mouse monoclonal antibody (MAb) specific to the vegetative cell of Clostridium butyricum strain MIYAIRI 588 (CBM588) was produced by immunisation with whole vegetative cells. This MAb (MAb-MS35) recognised a 35 KDa surface protein on CBM588 and did not cross-react with other intestinal bacteria including 16 strains of C. butyricum. A close correlation was observed between CBM588 vegetative cell counts in rat faeces and enzyme-linked immunosorbent assay (ELISA) values using MAb-MS35. Consequently, this ELISA was used to specifically determine the concentrations of CBM588 in the faeces of rats. The aim of this study was to investigate the fate of CBM588 in the rat gastrointestinal tract after a single oral administration. CBM588 spores were administered intragastrically at 107 cells per rat, and the faeces of each rat were collected twice daily. CBM588 vegetative cells were detected in the faeces for 3 d after administration. Total recovery of CBM588 vegetative cells from faeces were three to six times the spore dose. In addition, CBM588 vegetative cells were observed microscopically using faecal smears and an immunogold-silver staining method with MAb-MS35. These findings indicate that CBM588 spores germinate and the vegetative cells multiply in the gastrointestinal tract of rats.
C. g-(D,L)-Polyglutamic acid [g-(D,L)-PGA, or g-PGA] produced by Bacillus subtilis (natto) both on laboratory and pilot scale fermenter systems has been characterized. g-PGA in its free acid form (H +) is in-soluble in water. The salt forms of K + , Na + , NH 4 + , Ca 2+ , and Mg 2+ of g-polyglutamates are fully soluble in water. The structural characteristics of the salts of g-Polyglutamates (Na + , K + , NH 4 + , Ca 2+ , and Mg 2+) were determined with 1 H-and 13 C-NMR spectroscopy and FT-IR spectroscopy. The thermal properties were determined with thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC). The typical physical and chemical properties including pH-titration curve, concentration-dependent viscosity, temperature-dependent viscosity, and heavy metal binding properties with Pb 2+ , Cd 2+ , and Cu 2+ were also determined. The biological functionalities were partially characterized with in vivo feeding studies with broilers and egg-layers, and activation of GTF (Glucose tolerance factor) activity with rat 3T3-L1 cell cul-ture studies. g-(D,L)-Polyglutamic acid [g-(D,L)-PGA] appears to have five different conformations de-pending on the environmental conditions. They are the a-helix, b-sheet, helix-to-random coil transition, random-coil and enveloped aggregate. The conformational states, hydrogen bonding and polyanionic na-ture make g-PGA a versatile multi-functional biopolymer possessing many useful biological function-alities.
Aims: To conduct in vitro and in vivo assessments of the safety of two species of Bacillus, one of which, Bacillus subtilis, is in current use as a food supplement.
Methods and Results: Cultured cell lines, Caco-2, HEp-2 and the mucus-producing HT29-16E cell line, were used to evaluate adhesion, invasion and cytotoxicity. The Natto strain of B. subtilis was shown to be able to invade and lyse cells. Neither species was able to adhere significantly to any cell line. The Natto strain was also shown to form biofilms. No strain produced any of the known Bacillus enterotoxins. Disc-diffusion assays using a panel of antibiotics listed by the European Food Safety Authority (EFSA) showed that only Bacillus indicus carried resistance to clindamycin at a level above the minimum inhibitory concentration breakpoints set by the EFSA. In vivo assessments of acute and chronic dosing in guinea pigs and rabbits were made. No toxicity was observed in animals under these conditions.
Conclusions: Bacillus indicus and B. subtilis should be considered safe for oral use although the resistance of B. indicus to clindamycin requires further study.
Significance and Impact of the Study: The results support the use of B. subtilis and B. indicus strains as food supplements.
Die Biotechnologie begleitet den Menschen seit Jahrtausenden. Sie war lange eher unbeachtet empirisch immer weiter entwickelt worden und hat so stetig zur gesellschaftlichen Entwicklung des Menschen beigetragen. In Zeiten materieller Engpässe wurden in rasantem Tempo biotechnologische Prozesse entwickelt. Beispiele sind die Verfahren zur Herstellung von Aceton, Glycerin oder Butanol während des Ersten Weltkrieges und die Entwicklung der Antibiotika während des Zweiten Weltkrieges.
Biotechnology is one of the leading fields both in research and in industry. Tremendous effort has been made during the last two decades. However, biotechnology is much older than a few decades. Classical biotechnological products and processes are used from mankind over more than 5000 years. This article reflects the historical development of biotechnology — starting from the Egypt breweries over the production of indigo and saltpetre in the middle age to the discoveries of Pasteur, Fleming and Weizmann.
The value of exogenously supplied live bacteria for the maintenance of health in humans has been recognized both scientifically in the published literature and commercially in the availability of probiotic products. Although many bacteria characterized as probiotics are strains of Lactobacillus or Bifidobacterium, sporeforming bacteria, primarily of the genus Bacillus and related genera, have also been studied and commercialized as probiotics. This article reviews the characterization, efficacy, and safety of sporeformers used as probiotics.
Sourdough technology is widely used; it is employed in bread making and for the production of cakes. Sourdough is characterized by a complex microbial ecosystem, mainly represented by lactic acid bacteria and yeasts, whose fermentation confers to the resulting bread its characteristic features such as palatability and high sensory quality. Investigation of the microbial composition of sourdough is relevant in order to determine the potential activities of sourdough microorganisms. This review focuses on the role of the most important group of sourdough fermenting bacteria that consists of lactobacilli; species that belong to the Lactobacillus genus are the main responsible of flavor development, improvement of nutritional quality as well as stability over consecutive refreshments of sourdough. Lactobacilli also establish some durable microbial associations. An overview of the tools for monitoring predominant species is also reported.
Post-weaning diarrhoea syndrome (PWDS) of piglets is caused mainly by enterotoxigenic Escherichia coli (ETEC) strains. A new in-feed probiotic, LSP 122 (Alpharma), containing viable spores of Bacillus licheniformis was tested for its efficacy to control PWDS in piglets in a low health-status farm, using four groups with a total of 256 weaned piglets for a 28-day period. One group (negative control) was offered antimicrobial-free and probiotics-free fed, one group was offered feed supplemented with 10(6)viable spores of Bacillus toyoi (Toyocerin(R)) per gram of feed and two groups were offered feed supplemented with 10(6)and 10(7)viable spores of B. licheniformis per gram of feed, respectively, and were compared with regard to the appearance of clinical signs, mortality, weight gain and feed conversion. The results showed that all groups supplemented with probiotics exhibited a reduced incidence and severity of diarrhoea. Mortality in all probiotic supplemented pigs was significantly lower compared with the negative control group (P<0.05). The evaluation of the weight gain data, as well as feed conversion ratio, indicated that the three treated groups performed remarkably better than the negative control group (P<0.05) and the group receiving the high inclusion of LSP 122 performed better than the two other groups receiving probiotics (P<0.05). No ETEC strains were detected on day 22 in the high inclusion of LSP 122 and Toyocerin groups as compared with the untreated control. It was concluded that the high dosage schedule of LSP 122, providing 10(7)viable spores of B. licheniformis per g of feed, is a very useful agent for the control of PWDS due to ETEC.
Bacillus spp. ST13, isolated from human stool, was evaluated for siderophoregenic and probiotic qualities prior to its possible application for iron nutrition in humans and animals. It was tested for siderophore production in iron-limiting conditions and found to produce catecholate type of siderophore on the basis of high-performance liquid chromatography (HPLC), FT-IR, NMR, and mass spectra analysis. The isolate was screened for probiotic properties as per WHO and FAO guidelines. The strain ST13 can survive stomach acidity, bile salt and partially simulated gastrointestinal tract conditions. It was susceptible to most of the antibiotic tested and showed antimicrobial activity against enteric pathogens like Salmonella typhimurium, Streptococcus pyogenes, and Staphylococcus aureus. Strain ST13 showed close similarity with Bacillus subtilis using 16S r-RNA gene sequence analysis and biochemical characterization. The methanolic extract of ST13 siderophore was evaluated for DPPH radical scavenging activity, which showed 94.55 ± 0.9% of radical scavenging effect.
The use of succinylated β-lactoglobulin as a novel functional tablet excipient for the protection of probiotic bacteria against the adverse gastric conditions and their delivery in the intestine was studied. Tablets were produced by direct compression of a dry mixture of Bifidobacterium longum HA-135 and the tested excipient. The results showed that tablets made of native β-lg did not ensure cell survival while grafting carboxylic acid groups on the protein revealed to be an innovative method to create a gastroresistant matrix that could allow the survival of up to 10(8)CFU and 10(7)CFU after 1h and 2h gastric incubation, respectively. When compared to other polymers, succinylated β-lg promoted the best survival both upon compression and after simulated gastric passage. The proportion of succinylated β-lg in the formulation could be lowered to 60% without modifying the protective ability of the matrix. Additionally, the tablets proved to be stable over a period of 3 months when refrigerated. Succinylated β-lg tablets are an interesting vehicle for the protection of acid-sensitive bacteria during transit in the upper gastro-intestinal tract.
The aim of this study was to investigate whether consumption of Lactobacillus plantarum HEAL 9 (DSM 15312) and Lactobacillus paracasei 8700:2 (DSM 13434) could affect naturally acquired common cold infections in healthy subjects.
A randomised, parallel, double-blind placebo-controlled study was performed to investigate whether intake of this probiotic mixture could reduce the risk of common cold episodes, number of days with common cold symptoms, frequency and severity of symptoms, and cellular immune response in common cold infections. A total of 272 subjects were supplemented daily with either 10(9) cfu (colony forming units) of probiotics (N = 135) or control (N = 137) for a 12-week period.
The incidence of acquiring one or more common cold episode was reduced from 67% in the control group to 55% in the probiotic group (p < 0.05). Also, the number of days with common cold symptoms were significantly (p < 0.05) reduced from 8.6 days in the control group to 6.2 days, in the probiotic group, during the 12-week period. The total symptom score was reduced during the study period from a mean of 44.4 for the control group to 33.6 for the probiotic group. The reduction in pharyngeal symptoms was significant (p < 0.05). In addition, the proliferation of B lymphocytes was significantly counteracted in the probiotic group (p < 0.05) in comparison with the control group.
In conclusion, intake of the probiotic strains Lactobacillus plantarum HEAL 9 (DSM 15312) and Lactobacillus paracasei 8700:2 (DSM 13434) reduces the risk of acquiring common cold infections.
This study aimed to evaluate the safety of Clostridium butyricum and to investigate the effect of C. butyricum on mice ecosystem in the intestinal tract by way of examining the population of different microorganisms isolated from caecal contents. We firstly evaluated the safety of C. butyricum using acute toxicity test and Ames test. Then forty male BALB/c mice were divided into the following four treatment groups, each consisting of ten mice: normal group, low-dose group, medium-dose group and high-dose group. Caecal contents were removed aseptically, immediately placed into an anaerobic chamber, and dissolved in sterile pre-reduced PBS. The determination of Enterococcus spp., Enterobacter spp., Lactobacillus spp., Bifidobacterium spp. and Clostridium perfringens was analyzed by the spread plate method, cell morphologies and biochemical profiles. The results showed the oral maximum tolerated dose of C. butyricum was more than 10 g/kg body weight in mice and no mutagenicity judged by negative experimental results of Ames test. And in medium- and high-dose groups, the populations of Bifidobacterium spp. and Lactobacillus spp. increased in caecum, as well as the ratios of Bifidobacterium spp. and Lactobacillus spp. to Clostridium perfringens (P < 0.01) as compared with the normal group. This research showed the intake of C. butyricum significantly improved the ecosystem of the intestinal tract in BALB/c mice by increasing the amount of probiotics and reducing the populations of unwanted bacteria.
Three studies were conducted using Clostridium perfringens as an intestinal challenge to produce necrotic enteritis (NE). The studies consisted of two battery screening studies and one production study in floor pens, which were used to test the effect of the addition of Bacillus licheniformis (DSM 17236) spores at different doses and feeding periods in comparison to birds fed diets with subtherapeutic levels of virginiamycin (15 g/ton feed). In all three studies the use of B. licheniformis (1.6 x 10(6) - 8 x 10(7) CFUs/g) or virginiamycin (15 g/ton feed) showed no difference in effect with regard to feed conversion ratio, weight gain, NE lesion score, and NE mortality. In the two battery studies, both treatments showed a significantly decreased feed conversion ratio, increased weight gain, reduced NE lesion score, and NE-reduced mortality compared to the nonmedicated C. perfringens-challenged group. In general, none of the treatments performed as well as the no-challenge group. The present data indicate that the use of B. licheniformis spores as a probiotic or direct-fed microbial could be an alternative to adding medication to the feed to overcome NE under commercial-like conditions and could therefore be of direct use in preventing antibiotic-resistant pathogens in chickens.
Angiogenesis is required for wound healing and repair, but dysregulated angiogenesis is involved in gastrointestinal inflammation. Bacillus polyfermenticus (B.P.) is a probiotic bacterium clinically used for a variety of intestinal disorders in East Asia. Here we investigated the effect of B.P. on angiogenesis of human intestinal microvascular endothelial cells (HIMECs) and wound healing in intestinal mucosa. Exposure of HIMECs to the conditioned medium of B.P. cultures (B.P. CM) increased cell migration, permeability, and tube formation. Production of the proangiogenic cytokine IL-8 was increased by B.P. CM, and neutralizing antibodies against IL-8 or IL-8 receptor CXCR2 reduced tube formation as well as actin stress fiber formation. B.P. CM also increased NF-kappaB activation, and inhibitors of NF-kappaB suppressed B.P. CM-induced tube formation and IL-8 production. Furthermore, B.P. facilitated recovery of mice from colitis as shown by increased body weight and reduced rectal bleeding and histological severity. B.P. also increased angiogenesis and mouse IL-8 production in the mucosal layer. Collectively, these results show that B.P. increases angiogenesis of HIMECs in a NF-kappaB/IL-8/CXCR2-dependent manner. Moreover, B.P. promotes angiogenesis in the mucosa during recovery of mice from colitis, suggesting that this probiotic may be clinically used to facilitate intestinal wound healing.
In the present study intestinal delivery systems resistant to gastric juice, loaded with the probiotic bacteria Lactobacillus acidophilus LA14 and Bifidobacterium lactis BI07, were produced by the polyelectrolyte complexation. First, beads were prepared by the traditional extrusion method and nine formulations were developed using alginate as main carrier and the biopolymer, xanthan gum (XG), as hydrophilic retardant polymer or the cellulose derivative, cellulose acetate phthalate (CAP), as gastro-resistant polymer. The results showed that the incorporation of the 0.5% (w/v) of XG or the 1% (w/v) of CAP within the 3% (w/v) of alginate solution increased the survival of the probiotic bacteria in acid conditions from 63% of the freeze-dried bacteria up to 76%. Subsequently, these formula was used to prepare smaller microcapsules by means of an atomization device. Despite of the high viscosity of the biomass suspension, the spraying system produced spherical and non-aggregated microcapsules able to survive in harsh condition better than beads: the survival of the probiotic bacteria after acid incubation was 91%. The performance of the microcapsules in simulated gastric fluid (SGF) containing pepsin and in gut medium (GM) containing bile salts was excellent (viability>95%). Furthermore, the viability of probiotic bacteria was maintained after an incubation of 24h in GM. Finally, stability tests performed at 5 degrees C highlighted a bacterial viability of about 82% and 70% after 6 and 9 months, respectively.
Animal feed often contains probiotic Bacillus strains used as feed additives. Spores of the non-pathogenic B. cereus var. toyoi (product name Toyocerin) are used. Distinguishing between toxic wild-type Bacillus cereus strains and this probiotic strain is essential for evaluating the quality and risk of feed. Bacillus cereus CIP 5832 (product name Paciflor was used as probiotic strain until 2001. The properties of the two probiotic strains are quite similar. Differentiating between probiotic strains and wild-type B. cereus strains is not easy. ss-lactam antibiotics such as penicillin and cefamandole exhibit an inhibition zone in the agar diffusion test of probiotic B. cereus strains which are not seen for wild-type strains. Therefore, performing the agar diffusion test first may make sense before FT-IR testing. When randomly checking these strains by Fourier transform infrared spectroscopy (FT-IR), the probiotic B. cereus strains were separated from wild-type B. cereus/B. thuringiensis/B. mycoides/B. weihenstephanensis strains by means of hierarchical cluster analysis. The discriminatory information was contained in the spectral windows 3000-2800 cm(-1) ("fatty acid region"), 1200-900 cm(-1) ("carbohydrate region") and 900-700 cm(-1) ("fingerprint region"). It is concluded that FT-IR spectroscopy can be used for the rapid quality control and risk analysis of animal feed containing probiotic B. cereus strains.
The pharmacology, uses, dosages, safety, drug interactions, and contraindications of probiotics are reviewed.
Probiotics are live nonpathogenic microorganisms administered to improve microbial balance, particularly in the gastrointestinal tract. They consist of Saccharomyces boulardii yeast or lactic acid bacteria, such as Lactobacillus and Bifidobacterium species, and are regulated as dietary supplements and foods. Probiotics exert their beneficial effects through various mechanisms, including lowering intestinal pH, decreasing colonization and invasion by pathogenic organisms, and modifying the host immune response. Probiotic benefits associated with one species or strain do not necessarily hold true for others. The strongest evidence for the clinical effectiveness of probiotics has been in the treatment of acute diarrhea, most commonly due to rotavirus, and pouchitis. More research is needed to clarify the role of probiotics for preventing antibiotic-associated diarrhea, Clostridium difficile infection, travelers' diarrhea, irritable bowel syndrome, ulcerative colitis, Crohn's disease, and vulvovaginal candidiasis. There is no consensus about the minimum number of microorganisms that must be ingested to obtain a beneficial effect; however, a probiotic should typically contain several billion microorganisms to increase the chance that adequate gut colonization will occur. Probiotics are generally considered safe and well tolerated, with bloating and flatulence occurring most frequently. They should be used cautiously in patients who are critically ill or severely immunocompromised or those with central venous catheters since systemic infections may rarely occur. Bacteria-derived probiotics should be separated from antibiotics by at least two hours.
Probiotics have demonstrated efficacy in preventing and treating various medical conditions, particularly those involving the gastrointestinal tract. Data supporting their role in other conditions are often conflicting.
A wealth of data implicates that ErbB receptors have essential roles in tumor development. Probiotic bacteria are known to exert an anticancer activity in animal studies. Bacillus polyfermenticus (B.P.), a probiotic bacterium, has been clinically used for a variety of gastrointestinal disorders in East Asia. Here, we investigated the effect of B.P. on the growth of tumors and its putative mechanism of actions. Conditioned medium of B.P. cultures (B.P. CM) inhibited the growth of human colon cancer cells including HT-29, DLD-1 and Caco-2 cells. Moreover, B.P. CM suppressed colony formation of HT-29 cells cultured on soft agar and reduced carcinogen-induced colony formation of normal colonocytes. Furthermore, data from the mouse xenograft model of human colon cancer cells showed reduced tumor size in B.P. CM-injected mice when compared to E. coli conditioned medium-injected mice. Exposure of B.P. CM to HT-29 cells for 24 hr, 48 hr and 2 weeks reduced ErbB2 and ErbB3 protein expression as well as mRNA levels. Moreover, cyclin D1 expression that is required for ErbB-dependent cell transformation was decreased by B.P. CM. Furthermore, transcription factor E2F-1 that regulates cyclin D1 expression was also decreased by B.P. CM. These results show that B.P. inhibits tumor growth and its anticancer activity occurs, at least in part, through suppressing ErbB2 and ErbB3. Taken together, our study suggests that this probiotic may be clinically used as a prophylactic treatment to prevent colon cancer development.
Use of sourdough is of expanding interest for improvement of flavour, structure and stability of baked goods. Cereal fermentations also show significant potential in improvement and design of the nutritional quality and health effects of foods and ingredients. In addition to improving the sensory quality of whole grain, fibre-rich or gluten-free products, sourdough can also actively retard starch digestibility leading to low glycemic responses, modulate levels and bioaccessibility of bioactive compounds, and improve mineral bioavailability. Cereal fermentation may produce non-digestible polysaccharides, or modify accessibility of the grain fibre complex to gut microbiota. It has also been suggested that degradation of gluten may render bread better suitable for celiac persons. The changes in cereal matrix potentially leading to improved nutritional quality are numerous. They include acid production, suggested to retard starch digestibility, and to adjust pH to a range which favours the action of certain endogenous enzymes, thus changing the bioavailability pattern of minerals and phytochemicals. This is especially beneficial in products rich in bran to deliver minerals and potentially protective compounds in the blood circulation. The action of enzymes during fermentation also causes hydrolysis and solubilisation of grain macromolecules, such as proteins and cell wall polysaccharides. This changes product texture, which may affect nutrient and non-nutrient absorption. New bioactive compounds, such as prebiotic oligosaccharides or other metabolites, may also be formed in cereal fermentations.
Although human consumption of Bacillus polyfermenticus provides several health benefits, the probiotic effect of this bacterium against colonic inflammation has not yet, to our knowledge, been studied. Therefore, we induced colitis in mice by oral or intrarectal administration of dextran sodium sulfate (DSS) or trinitrobenzenosulfonic acid (TNBS), respectively, and investigated the effect of B. polyfermenticus on colitis. We found that mice treated with DSS or TNBS along with B. polyfermenticus had reduced mortality and severity of colitis (weight loss, diarrhea, and mucosal damages) than mice treated with DSS or TNBS alone. B. polyfermenticus also reduced the expression of inflammatory molecules, including chemokine (C-X-C motif) ligand 1, intercellular adhesion molecule, and tumor necrosis factor-alpha, but enhanced the expression of the antiinflammatory cytokine interleukin-10 in the inflamed mouse colon. Moreover, B. polyfermenticus suppressed apoptosis both in vivo in inflamed colonic mucosa and in vitro in colonic epithelial cells stimulated with apoptosis-inducing agents (FasL or Clostridium difficile Toxin A) when the apoptotic response was determined by a terminal deoxynucleotidyl transferase dUTP nick end labeling assay and cleavage of poly(ADP-ribose) polymerase or caspase-3, respectively. Treating colonic epithelial cells with B. polyfermenticus-conditioned medium (BPCM) enhanced cell proliferation and induced the phosphoinositide 3-kinases/Akt signaling pathway, suggesting that this bacterium can promote epithelial cell proliferation. BPCM also promoted the migration of colonic epithelial cells. These data suggest that B. polyfermenticus ameliorates colonic inflammation by suppressing apoptosis and promoting epithelial cell proliferation and migration.
To evaluate the therapeutic effects of a probiotic supplement (Clostridium butyricum, CGMCC0313) in a chemically-induced rat model of experimental colitis.
An experimental ulcerative colitis model was established by rectal injection of oxazolone into the colon of 40 Wistar rats randomly divided into four groups. The positive control group was sacrificed 3 d after colitis onset. The remaining groups were fed daily with either 2 mL of C. butyricum (2.3 x 10(11) CFU/L), 2 mL of mesalamine (100 g/L), or 1 mL of sodium butyrate (50 mmol/L) for 21 d. The animals' body weight, behavior, and bowel movements were recorded weekly. After sacrifice, visual and microscopic observations of pathological changes of colon tissue were made, body weight and wet colon mass index were measured and recorded, and serum levels of interleukin-23 (IL-23) and TNF-alpha were measured using ELISA. Expression of calcitonin gene-related peptide in colon tissue was measured by RT-PCR. Finally, changes in rat intestinal microflora status were measured in all groups.
We found that treatment with C. butyricum lowered the serum levels of both IL-23 and tumor necrosis factor-alpha (TNF-alpha) with similar or even better efficiency than that of mesalamine or sodium butyrate. The rat intestinal flora appeared to recover more quickly in the group treated with C. butyricum than in the mesalamine and sodium butyrate groups. Finally, we found that the expression level of calcitonin gene related peptide was elevated in colon tissue in the sodium butyrate treated group but not in the C. butyricum or mesalamine treated groups, indicating a sensitization of colon following sodium butyrate treatment.
In our experimental colitis model, treatment with C. butyricum CGMCC0313, a probiotic supplement, is at least as efficient as treatment with mesalamine.
Siderophoregenic Bacillus strain DET9 has been selectively isolated from dairy waste. It was evaluated for probiotic characteristics and susceptibility pattern against antibiotics. Its spores showed excellent tolerance to simulated gastrointestinal tract conditions and exhibited antimicrobial activity against organisms such as Escherichia coli, Micrococcus flavus, and Staphylococcus aureus. Its susceptibility to antibiotics reduces the prospect to donate resistance determinants if administered in the form of probiotic preparations. It was observed to produce approximately 60 mg/l catecholate type of siderophore under iron stressed conditions, identified as a 2,3-dihydroxy benzoic acid by high-performance liquid chromatography, infrared spectroscopy, nuclear magnetic resonance, and mass spectral analysis. Partial 16S-rRNA gene sequencing analysis shows that the isolate exhibited homology with Bacillus thuringiensis and Bacillus weihenstephanensis, whereas biochemical characterization revealed its novelty. DET9 exhibited no mortality of fishes in a 60-day trial, when fishes (surfi tetra) were challenged up to 100 ppm cell concentration, with their daily diet.
The work aims to provide evidence on the viability of Lactobacillus spp. and a spore form of Bacillus subtilis from nonprocessed bacteria to coated dosage forms (i.e., mini-tablets, pellets, and their coated forms). Lactobacillus spp. were cultivated overnight in MRS broth (10(9) cfu/mL) and B. subtilis spores were produced on plate count agar (10(7) cfu/mL) for 2 weeks. Bacteria and spores were freeze-dried in skim milk enriched with glycerol. The cakes were further processed into tablets (2.5 mm diameter) by direct compression with or without microcrystalline cellulose and inulin. Pellets (1-1.4 mm diameter) were produced by extrusion-spheronization of bacterial and spore suspensions with microcrystalline cellulose, lactose, inulin, and skim milk. Both tablets and pellets were film coated. The properties of the dosage forms, particularly the bacterial viability, were evaluated immediately after production and throughout storage for 6 months at 4 degrees C. The study has shown that for an adequate stabilization of the bacteria a protective matrix (e.g., skim milk) and cryoprotectors (e.g., glycerol) must be present at early stages of bacterial de-hydration. Tabletting had a less deleterious effect (<2 log units) on bacteria when compared to pelletization (in some cases 3 log units). Enteric coating (15%, w/w) of either tablets or pellets did not affect the viability of the bacteria.
To characterize the antimicrobial and adhesion ability of candidate probiotic Clostridium butyricum CB2 for farmed fish in vitro.
The potential probiotic Cl. butyricum CB2 had been evaluated for its adhesion capacity and antagonistic effect against two fish pathogens Aeromonas hydrophila and Vibrio anguillarum by the intestinal cell model. In addition, the aggregation ability and antimicrobial property on agar plate were assayed. The results indicated that the candidate probiotic Cl. butyricum CB2 have strong adhesion property and a higher antagonistic activity to Aer. hydrophila and V. anguillarum both on agar plate and cell model. Clostridium butyricum showed a higher aggregation which might be the reasons for bacteria adhesion and antimicrobial activity.
The strain Cl. butyricum CB2 could be used as potential probiotic to inhibit pathogens growth and prevent their colonization in fish intestinal tract.
This study revealed the antimicrobial and adhesion characteristic of Cl. butyricum CB2 which was selected as the potential probiotic to farmed fish.