ArticleLiterature Review

Spore-forming bacteria and their utilisation as probiotics

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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.

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... In this study, the application of a PF-based dietary pressure to modulate European sea bass gut microbiota composition and corresponding metabolic functions revealed to be a successful strategy to find carbohydrate-active bacteria with probiotic potential. In particular, we targeted and isolated spore-forming Bacilli, commonly used in probiotic preparations, mainly due to their extreme resistance characteristics and indefinitely survival, advantageous for industrial applications 29,32,34,38 . ...
... For instance, determining the number and type of CAZymes present in each genome will provide deeper understanding on their carbohydrolytic potential but might also allow identification of genomic features responsible for adaptation to life within the gut that may support the role of Bacillus spp. as probiotics 39,[71][72][73] . The growing applications of spores in biomedicine and biotechnology (as oral vaccines, probiotics or display systems) [32][33][34][35]38,74 , and the fact that there are approximately 30 probiotic strains approved as feed additives in European Union, but only one for aquaculture (Bactocell ® , which is not a sporeformer formulation), underscore the importance of this study. ...
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The gastrointestinal microbiota plays a critical role on host health and metabolism. This is particularly important in teleost nutrition, because fish do not possess some of the necessary enzymes to cope with the dietary challenges of aquaculture production. A main difficulty within fish nutrition is its dependence on fish meal, an unsustainable commodity and a source of organic pollutants. The most obvious sustainable alternatives to fish meal are plant feedstuffs, but their nutritive value is limited by the presence of high levels of non-starch polysaccharides (NSP), which are not metabolized by fish. The composition of fish-gut microbial communities have been demonstrated to adapt when the host is fed different ingredients. Thus, we hypothesized that a selective pressure of plant-based diets on fish gut microbiota, could be a beneficial strategy for an enrichment of bacteria with a secretome able to mobilize dietary NSP. By targeting bacterial sporulating isolates with diverse carbohydrase activities from the gut of European sea bass, we have obtained isolates with high probiotic potential. By inferring the adaptive fitness to the fish gut and the amenability to industrial processing, we identified the best two candidates to become industrially valuable probiotics. This potential was confirmed in vivo, since one of the select isolates lead to a better growth and feed utilization efficiency in fish fed probiotic-supplemented plant-based diets, thus contributing for sustainable and more cost-effective aquaculture practices.
... 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]. ...
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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.
... Probiotics are last recent biological methods by using alive and suitable microorganisms that can beneficial effects to the host health when consumed in appropriate and regular quantities [6] . This microorganisms have been used to improve the in vitro fermentation characteristics of the roughages with low-quality [7] The scientific interest to several bacillus strains have been screened for their potential probiotic functionalities with special regard to Bacillus natto and Bacillus subtilis due to their beneficial effects on ruminants [8] . However, these species were firstly used as probiotics product (Enterogermina®) in Italy since 1958 [9] . ...
... After that, 10 mL of preservation solutions were diluted to 100 mL with distilled water as the working solution in which the concentration of N was 10 mg/dL. Subsequently, 0, 1, 2, 4, and 6 mL of working solutions were separately mixed with 10,9,8,6, and 4 mL of distilled water and then all diluted with 0.2 M HCl to 50 mL as the NH 4 Cl standard solutions in which the concentrations of N were 0, 0.2, 0.4, 0.8 and 1.2 mg/dL, respectively. Finally, 0.4 mL of the NH 4 Cl standard solution of each concentration was treated to obtain a standard curve as previously mentioned. ...
Article
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The study was carried out to investigate the impact of Bacillus coagulans supplementation with different concenteration (0, 0.25×107 , 0.50×107 , and 0.75×107 cfu/mL) on in vitro parameters as methane (CH4) parameters, nutrients digestibility and the fermentation character of rumen of fibrous agriculture by-products (maize stover and rice straw) using probiotics preparations in ruminants. The results showed that, maximum gas production (Vf), gas production fraction (k) and the time when half of the maximum gas production was achieved (t0.5) of maize stover were significantly increased when compared to group of the rice straw. Additionally, in vitro dry matter disappearance; (IVDMD), in vitro neutral detergent fiber disappearance (IVNDFD), individual volatile fatty acids (VFAs) and total VFA (TVFA) of maize stover were significantly in related to the rice straw group. Also, the gas production rate at the early incubation stage (FRD0), pH value of rumine and the acetate: propionate ratio of maize stover is significantly lower than the rice straw. Vf of rice straw was quadratic (P
... 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]. ...
Article
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.
... Certain strains of Bacillus are probiotic bacteria, owing to a myriad of benefits they provide for body metabolism [9,10]. Probiotic Bacillus produces a variety of enzymes, such as protease, amylase, and lipase [11][12][13], which are beneficial to animal digestion and the development of the small intestinal micro-morphological structure [12,14]. Bacillus also responds to improving the bacterial composition of the gut microbiota [15,16]. ...
... * means significant difference at 0.05 levels (P < 0.05), and ** means significant difference at 0.01 levels (P < 0.01) affect body metabolism [24,46]. A balanced and optimized intestinal microbiota is critical to improve body growth and keep health, especially to neonatal chicks [11,47]. The particular physiological character of chicks is the lack of colon, which make the roles of cecum more important than in other animals and human being [48]. ...
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The aim of our study was to unveil the promoting function of compound Bacillus sp. in improving chicken digestion-induced higher body growth performance. Strains of Bacillus subtilis and B. licheniformis were jointly supplemented to the chick diet. Digestive enzyme activities in the digesta improved, development of intestinal villus enhanced, and duodenum and ileum villous height increased, while their crypt depth declined, and the cecum’s bacterial composition optimized after 56 days of supplementation. Bacterial composition at the phylum level changed significantly, more Firmicutes, Proteobacteria, Epsilonbacteraeota, and Tenericutes, but fewer Bacteroidetes were detected in cecum digesta in the compound Bacillus supplemented group. Bacterial composition diversity, which improves the abundances of metabolic genes through KEGG pathway classification, became more abundant. Results indicated that the Ruminococcaceae UCG-005, unclassified Ruminococcaceae, and unclassified Lachnospiraceae species are actively correlated with body growth, promoting higher final body weight. In conclusion, owing to digestive enzyme secretion, the development of intestinal villus was stimulated and gastrointestinal bacterial composition optimized, and two combined Bacillus sp. improved chicken body growth. Our findings show the promoting action of Bacillus subtilis and B. licheniformis on digestion, which can be an alternative to antibiotics.
... Bacillus licheniformis has a strong ability to produce protease, lipase, and amylase, which facilitates an efficient degradation of feed for nutrient absorption (Rozs et al. 2001). B. subtilis and B. licheniformis are the two kinds of probiotics that can be supplemented in animal diet (Bader et al. 2012) and the combined use of these two Bacillus species might play a significant role in poultry growth. Multi-strain probiotics have been found to be more effective than single-strain probiotics (Chapman et al. 2011). ...
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A total of 900 one-d-old Chinese Huainan Partridge Shank chickens were randomly allocated into nine groups with five replicates of 20 each. Birds were fed with basal diet, basal diet supplemented with 150 mg kg⁻¹ aureomycin, basal diet supplemented with different proportions of Bacillus licheniformis and Bacillus subtilis, which was 0:1.0×10⁶, 2.5×10⁵:7.5×10⁵, 3.3×10⁵:6.6×10⁵, 5.0×10⁵:5.0×10⁵, 6.6×10⁵:3.3×10⁵, 7.5×10⁵:2.5×10⁵ and 1.0×10⁶:0, respectively. The duration of the experiment was 56 d. The results indicated that dietary supplementation of 6.6×10⁵:3.3×10⁵ of B. lichenifornis:B. subtilis improved final body weight, increased the average daily gain, and reduced feed/gain ratio (P<0.05). The numbers of total Lactobacillus and Bifidobacterium sp. in the caecum significantly increased, and the numbers of Escherichia coli and Salmonella sp. significantly declined compared to that of the control (P<0.05). Intestinal villous height and villous height to crypt depth ratio of the duodenum, jejunum, and ileum were significantly higher than that of the control, and intestinal crypt depth of the duodenum and ileum was significantly lower (P<0.05). The total antioxidant capacity, total superoxide dismutase, and glutathione peroxidase ability in plasma significantly improved, while the malondialdehyde concentration in plasma decreased (P<0.05). Compared to the control, plasma concentrations of ammonia, uric acid and urea nitrogen and the activity of xanthine oxidase were reduced (P<0.05). In conclusion, an inclusion of 6.6×10⁵:3.3×10⁵ of B. licheniformis: B. subtilis to the diet improved the growth performance, caecal microbiota, plasma biochemical profile, and significantly improved the small intestine morphology, while reducing the mortality rate.
... strains have been suggested as feed supplements for both, human beings and other animals [9][10][11]. Recent evidence suggests that Bacillus probiotics, which germinate within the gastrointestinal tract of poultry [5,12], may form a small population within the normal microbiota [13]. In addition, probiotic Bacillus produce a variety of enzymes, such as protease, amylase, and lipase [14,15]. ...
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Sixty-week-old Hy-Line brown laying hens were randomly divided into five groups and fed different diets over a period of 84 days. Experimental treatments included a basal diet (control); the basal diet supplemented with 1.0 × 10⁶B. licheniformis yb-214245; the basal diet supplemented with 1.0 × 10⁶B. subtilis yb-114246; a combination of both strains in a 2:1 ratio (6.6 × 10⁵:3.3 × 10⁵B. licheniformis yb-214245:B. subtilis yb-114246); and the latter, added with 5 mg/kg flavomycin. Basal diet supplementation with the combined Bacillus species improved egg-laying performance in aging hens significantly (P < 0.05). Eggshell strength improved significantly with this treatment, compared to the control or the antibiotic-supplemented groups (P < 0.05). The levels of total cholesterol, triglycerides, and very low-density lipoprotein cholesterol in egg yolk declined significantly more in the Bacillus-treated group than in the control or the antibiotic-supplemented groups (P < 0.01). Small intestinal morphology was better in the hens treated with the Bacillus combination than in the hens in the control group (P < 0.05). The total number of aerobic bacteria (Bacillus, Lactobacillus, and Bifidobacterium) in the cecum was significantly higher in all the Bacillus-supplemented hens than either in the control or the antibiotic-supplemented hens (P < 0.01); additionally, the number of E. coli and Salmonella was significantly lower than in the control group (P < 0.01). In conclusion, diet supplementation with the combination of Bacillus species used here for aging laying hens improved their growth performance, cecal bacterial composition, egg quality, and small intestine morphology.
... До складу біоспорину входить 2 види бацил -Bacillus subtilis та Bacillus licheniformis. Як і попередні препарати, цей пробіотик має високу антагоністичну активність щодо умовно-патогенних і патогенних мікроорганізмів людського організму, не виявляючи негативного ефекту на представників природної мікрофлори [12,13,20]. Крім того, завдяки синтезу бактеріями, які входять до складу біоспорину комплексу ферментів, відбувається стимуляція і регулювання нормального травлення, поліпшення всмоктування в шлунково-кишковому тракті вітамінів Е, Д і нормалізація продукції вітамінів групи В. Разом з тим препарат позитивно впливає на імунну систему: активує макрофаги, стимулює реплікацію імуноглобулінів, підсилює вироблення ендогенного інтерферону та лізоциму. ...
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Introduction. When it is nessecery to apply to the antimicrobial elimination therapy, the application of appropriate biological drugs is thealternative treatment. One of them is Biosporin (includes Bacillus subtilis та Bacillus licheniformis). It has a high antagonistic activity against pathogenic and opportunistic pathogenic microorganisms and expresse the immunomodulatory effect. The purpose of our work was to investigate the effect of probiotic Biosporin on different representatives of intestinal microbiota and confirm its activity against opportunistic microorganisms while maintaining the normal symbionts of intestine. Materials and methods. Experiments were conducted in the albino Wistar rats (males) with initial weighing 180-210 g, which were kept under standard vivarium conditions as required by ethics under the provisions of the European Convention for the protection of vertebrate animals used for experimental and other scientific purposes. Animals were in vivarium under appropriate lighting conditions, temperature and standard diet. During the study they had free access to water. The animals were divided into two groups: 1) control group (n = 12), 2) researched (n = 12). The first group was formed with intact rats. Animals of the second group were given the biological preparation Biosporin in a daily dose 0,2-0,3ml / animal per day. Calculation of doses were performed using doses of individual animals to humans. The biological preparation was administered intragastric every day for 10 days using nontraumatic probe. Faeces were collected on the 10th day into sterile container. When conducting microbiological investigations classic cultural method and differential diagnostic and special nutrient media for the cultivation of certain genera of microorganisms were used. The presence and quantitative values of enterococci, Escherichia, conditionally – pathogenic microorganisms, microaerophilic bacteria – lactobacilli and bifidobacteria were revealed. The results obtained in the investigations of intestinal microflora of experimental rats with the same ones in intact animals were compared. Number of the members of the intestinal microbial landscape was determined by counting of the colonies that grew on media in terms of 1g weight of feces.The conversion to lg of CFU /g (logarithm of the colony forming units in 1 g of faeces) was carry out. The identification of isolated cultures was carried out by morphological, tinctorial, cultural and biochemical properties. State microbiota of the colon was assessed by the population levels of certain species (groups) of microorganisms. Statistical analysis of the results was performed by the method of variational statistic with the determination of the average values of the quantities and the average error. Assessment of the reliability of the differences between average values during the analysis were obtained using unpaired t-Student test. The difference between the values was considered reliable when probability difference p ≤ 0,05. Results and discussion. Giving the Biosporin to the healthy animals slightly affected the studied quantitative microbial symbionts. Thus, the level of representatives of coccal group (staphylococci and streptococci) was partly higher compared with the intact animals. Staphylococci were found in the number of (5,6 ± 0,48) in experimental animals to (5,2 ± 0,35) lg CFU/ g in control group. Deviation of enterococci compared with control were more significant (9,6 ± 0,60 to 7,0 ± 0,46 lgCFU / g). Streptococci, which do not belong to the genus Enterococcus, were not found in the feaces of intact animals, but after given to rats Biosporin, were found in small amounts (1,2 ± 0,2 lgCFU / g). The highest rate was in Escherichia. These enterobacteria were found in the experiment at 6,35 ± 0,57 lg CFU / g to 5.9 ± 0.40 in control. The absence of Proteus under the influence of Biosporin was noticed, at the time, Proteus mirabilis in the amounts 6,0 ± 0,58 in intact animals was found. Microaerophilic microorganisms (Lactobacillus) in experiment and in intact animals at 7.0 were found out. On the part of bifidoflora some of their elimination, accompanied by a decrease of bifidobacteria compared with controls from 7,5 ± 0,76 to 6,75 ± 0,72 lg CFU / g was recorded. Changes of the amount of yeast fungi Candida in both groups were not recorded.The results of the research of the influence of Biosporin on faecal microbiota of healthy rats is an evidence that the drug causes some redistribution of microbial symbionts. It is clear that in the presence of bacteria – antagonist in the organism of rat the proliferation of investigated microorganisms is unlikely. Increasing of the number of enterococci and streptococci appearance is the result of the movement of microbes from the upper digestive tract, where they are more at its distal. A similar phenomenon also explain an activation of Escherichia in faeces. The number of bifidobacteria does not go beyond the norm. Thus, under conditions of a healthy body Biosporin initiates translocation of symbionts from proximal to distal digestive tract. Moreover, the discharge of natural biological niches promotes renewal of the population. With an excess of opportunistic flora, which includes representatives of the genus Proteus, Biosporin causes their elimination, reflected an appreciable therapeutic effect. Conclusions. The results of investigations of Biosporin influence on faecal microbiota indicate a redistribution of microbial symbionts. The increase of enterococci in the number and some activation of Escherichiain colon was established. Changes of lactobacilli and bifidobacteria number are insignificant. Obviously, the positive effect of Biosporin is more prominent in the case of dysbiotic changes (excessive proliferation of microflora). The proclivity of quantitative glycolytic microaerophilic microflora decreasing of Biosporin action, is the reason to use the probiotic preparations for reimplantation of lack bifidobacteria and lactobacilli as the next step after Biosporin utilisation.
... There has been recent interest in utilizing Clostridia as probiotics and there is potential for considering these organisms as probiotics, but they could be difficult to produce since they are anaerobes. Clostridia are already on the market as probiotics for human gastrointestinal applications, so this is an opportunity for new investigations into potential use of clostridia as probiotics during foodanimal production [82]. Finally, the in ovo methodology can be adopted to supply the chicken embryo with additional nutrients prior to hatching which will continue to be utilized by the chick post-hatch during the fasting period. ...
Article
Due to the continuing global concerns involving antibiotic resistance, there is a need for scientific forums to assess advancements in the development of antimicrobials and their alternatives that might reduce development and spread of antibiotic resistance among bacterial pathogens. The objectives of the 2nd International Symposium on Alternatives to Antibiotics were to highlight promising research results and novel technologies that can provide alternatives to antibiotics for use in animal health and production, assess challenges associated with their authorization and commercialization for use, and provide actionable strategies to support their development. The session on microbial-derived products was directed at presenting novel technologies that included exploiting CRISPR-Cas nucleases to produce sequence-specific antimicrobials, probiotics development via fecal microbiome transplants among monogastric production animals such as chickens and mining microbial sources such as bacteria or yeast to identify new antimicrobial compounds. Other research has included continuing development of antimicrobial peptides such as newly discovered bacteriocins as alternatives to antibiotics, use of bacteriophages accompanied by development of unique lytic proteins with specific cell-wall binding domains and novel approaches such as microbial-ecology guided discovery of anti-biofilm compounds discovered in marine environments. The symposium was held at the Headquarters of the World Organisation for Animal Health (OIE) in Paris, France during 12–15 December 2016.
... There has been recent interest in utilizing Clostridia as probiotics and there is potential for considering these organisms as probiotics, but they could be difficult to produce since they are anaerobes. Clostridia are already on the market as probiotics for human gastrointestinal applications, so this is an opportunity for new investigations into potential use of clostridia as probiotics during foodanimal production [82]. Finally, the in ovo methodology can be adopted to supply the chicken embryo with additional nutrients prior to hatching which will continue to be utilized by the chick post-hatch during the fasting period. ...
Article
Full-text available
Due to the continuing global concerns involving antibiotic resistance, there is a need for scientific forums to assess advancements in the development of antimicrobials and their alternatives that might reduce development and spread of antibiotic resistance among bacterial pathogens. The objectives of the 2nd International Symposium on Alternatives to Antibiotics were to highlight promising research results and novel technologies that can provide alternatives to antibiotics for use in animal health and production, assess challenges associated with their authoriza- tion and commercialization for use, and provide actionable strategies to support their development. The session on microbial-derived products was directed at presenting novel technologies that included exploiting CRISPR-Cas nucle- ases to produce sequence-specific antimicrobials, probiotics development via fecal microbiome transplants among monogastric production animals such as chickens and mining microbial sources such as bacteria or yeast to identify new antimicrobial compounds. Other research has included continuing development of antimicrobial peptides such as newly discovered bacteriocins as alternatives to antibiotics, use of bacteriophages accompanied by development of unique lytic proteins with specific cell-wall binding domains and novel approaches such as microbial-ecology guided discovery of anti-biofilm compounds discovered in marine environments. The symposium was held at the Headquarters of the World Organization for Animal Health (OIE) in Paris, France during 12–15 December 2016.
... The use of bacterial spore formers, Bacillus spp. in particular, as probiotics could provide advantages for both humans and animals (Cutting, 2011). The ability to form spores endows probiotics with higher resistance to technological stresses during production and storage processes and also higher resistance to gastric (pH, digestive enzymes) and intestinal environmental conditions (Bader et al., 2012). Indeed, a number of products containing spores of Bacillus spp. ...
Article
An experiment was conducted for the 56 d period in thirty six adult male albino Wister rats divided equally into four groups. After 7 d adaptation period, all the groups were fed with cholesterol enriched diet for 14 d. Thereafter, one group (T1) was fed on sterile skim milk along with cholesterolemic diet for the next 21 d. The two experimental groups (T2 and T3) received cholesterolemic diet plus spore biomass of B. coagulans B37 and B. pumilus B9, respectively, suspended in sterilized skim milk @ 8-9 log cfu/ ml for the next 21 d. The control group was supplied with clean water along with cholesterolemic diet for 21 d. All the animals were fed on the basal diet along with water during a 14 d post-treatment period. After the adaptation period, the blood samples were collected weekly from the experimental rats up to 42 d, and plasma separated, and subjected to the determination of different plasma lipid fractions. The oral administration of both B. coagulans B37 and B. pumilus B9 resulted a decrease (P Less Than 0.05) in plasma cholesterol, low density lipoprotein (LDL)-cholesterol concentrations and atherogenic index in the experimental rats.
... Several butyrate-producing strains have been isolated from chicken caecal content [14], however only a minority have been characterized and are spore-forming bacteria [15][16][17]. Utilization of spores as probiotics is advantageous because of their higher resistance to acidic conditions in the stomach, and the requirement of smaller numbers compared to probiotic vegetative cells [18]. In this study spore-forming butyrate-producing strains with 99 % 16S rRNA gene sequence similarity to each other, obtained from chicken caecal content is characterized in more detail. ...
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Strains of a Gram-stain-negative, rod-shaped and immotile bacterium were isolated from broiler chicken caecal content. The isolates required strict anaerobic conditions for growth, formed spores, were catalase-positive and oxidase-negative. They produced butyrate as the major metabolic end product in reinforced clostridial medium broth. The genomic DNA G+C content of the isolated strains was 32.5-34.6 mol%. The major cellular fatty acids were C16 : 0 FAME, C14 : 0 FAME, C19 : 0CYC 9,10DMA and C16 : 0DMA. The fatty acid composition of the cell wall showed no similarity to any strain in the midi database. 16S rRNA gene sequence analysis showed that the nearest phylogenetic neighbours were Anaerostipes hadrus and Clostridium populeti (92 % sequence similarity) within Clostridium cluster XIVa of the phylum Firmicutes. Therefore, a novel genus is proposed, with the name Caecibacterium sporoformans gen. nov., sp. nov. The type strain of Caecibacterium sporoformans is LMG 27730(T)=DSM 26959(T).
... Besides qualifying the mandatory bench marks for a candidate probiotic; Bacillus spp. offers higher acid tolerance and better stability during heat processing and low temperature storage (Bader et al., 2012). Additionally, they have also been shown to possess pathogen exclusion, anti-oxidant, antimicrobial, immuno-modulatory (Lefevre et al., 2015;Shobharani et al., 2015;Ripert et al., 2016) and food fermentation (Terlabie et al., 2006) abilities. ...
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Spore-forming bacilli are being explored for the production and preservation of food for many centuries. The inherent ability of production of large number of secretory proteins, enzymes, antimicrobial compounds, vitamins, and carotenoids specifies the importance of bacilli in food chain. Additionally, Bacillus spp. are gaining interest in human health related functional food research coupled with their enhanced tolerance and survivability under hostile environment of gastrointestinal tract. Besides, bacilli are more stable during processing and storage of food and pharmaceutical preparations, making them more suitable candidate for health promoting formulations. Further, Bacillus strains also possess biotherapeutic potential which is connected with their ability to interact with the internal milieu of the host by producing variety of antimicrobial peptides and small extracellular effector molecules. Nonetheless, with proposed scientific evidences, commercial probiotic supplements, and functional foods comprising of Bacillus spp. had not gained much credential in general population, since the debate over probiotic vs pathogen tag of Bacillus in the research and production terrains is confusing consumers. Hence, it’s important to clearly understand the phenotypic and genotypic characteristics of selective beneficial Bacillus spp. and their substantiation with those having GRAS status, to reach a consensus over the same. This review highlights the probiotic candidature of spore forming Bacillus spp. and presents an overview of the proposed health benefits, including application in food and pharmaceutical industry. Moreover, the growing need to evaluate the safety of individual Bacillus strains as well as species on a case by case basis and necessity of more profound analysis for the selection and identification of Bacillus probiotic candidates are also taken into consideration.
... An industrial application for clostridial spores is the use of C. butyricum spores as a probiotic (127). A general advantage of sporeformers as probiotics is that the number of spores added to food can be reduced by about 100-fold in comparison with vegetative cells. ...
Article
Physiology and Sporulation in Clostridium, Page 1 of 2 Abstract Clostridia are Gram-positive, anaerobic, endospore-forming bacteria, incapable of dissimilatory sulfate reduction. Comprising approximately 180 species, the genus Clostridium is one of the largest bacterial genera. Physiology is mostly devoted to acid production. Numerous pathways are known, such as the homoacetate fermentation by acetogens, the propionate fermentation by Clostridium propionicum, and the butyrate/butanol fermentation by C. acetobutylicum, a well-known solvent producer. Clostridia degrade sugars, alcohols, amino acids, purines, pyrimidines, and polymers such as starch and cellulose. Energy conservation can be performed by substrate-level phosphorylation as well as by the generation of ion gradients. Endospore formation resembles the mechanism elucidated in Bacillus. Morphology, contents, and properties of spores are very similar to bacilli endospores. Sporulating clostridia usually form swollen mother cells and accumulate the storage substance granulose. However, clostridial sporulation differs by not employing the so-called phosphorelay. Initiation starts by direct phosphorylation of the master regulator Spo0A. The cascade of sporulation-specific sigma factors is again identical to what is known from Bacillus. The onset of sporulation is coupled in some species to either solvent (acetone, butanol) or toxin (e.g., C. perfringens enterotoxin) formation. The germination of spores is often induced by various amino acids, often in combination with phosphate and sodium ions. In medical applications, C. butyricum spores are used as a C. difficile prophylaxis and as treatment against diarrhea. Recombinant spores are currently under investigation and testing as antitumor agents, because they germinate only in hypoxic tissues (i.e., tumor tissue), allowing precise targeting and direct killing of tumor cells.
... In this study we tested the hypothesis that there is a proportional link between bacterial probiotic characteristics and their yielded levan. There is increased interest from a research and commercial point of view in using spore-forming bacteria as probiotics (Foligne et al. 2012;Bader et al. 2012). These bacteria possess several attractive features such as thermo-stability, resistance to harsh environments, and permitting prolonged storage periods without any preparation. ...
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Five bacterial isolates from honey and bee gut were selected based on their high levansucrase activity and levan yield which were strongly positively correlated. All isolates showed good tolerance to temperature up to 70 °C, to NaCl up to 3 M and to 0.1% H2O2. They maintained over 59 and 64% survival at pH 9.0 and 2.0 respectively, but showed varying tolerance to 0.1% bile salts and pancreatic enzymes. Most isolates were susceptible to widely used antibiotics, but demonstrated diverse antimicrobial activity. Non hemolytic isolates were identified on the basis of 16S rRNA sequencing as Bacillus subtilis HMNig-2 and B. subtilis MENO2 with 97% homology. They exhibited promising probiotic characteristics and achieved highest levansucrase activity of 94.1 and 81.5 U/mL respectively. Both exhibited highest biofilm formation ability in static microtiter plate assay. Also, they achieved 34 and 26% adhesion respectively to Caco-2cells and had highest free radical scavenging activity of 30.8 and 26.2% respectively. The levans of the two isolates showed good antimicrobial activity against some pathogens and exhibited positive prebiotic effect (prebiotic index >1) with Lactobacillus casei and Lactobacillus reuteri. Results suggest a correlation between levansucrase production, levan yield and pre-probiotic activities of the studied strains.
... The merits of Se and probiotic bacteria on body were well-documented [19,24]. B. licheniformis and B. subtilis, two strains of probiotic Bacillus, have a better ability to live in adverse environments (low-moisture, high pelleting temperature, and less nutrients) than Lactobacillus, [25], which were more widely developed as additives in the biomedical industry. SECB owing the organic Se and compound Bacillus (B. ...
Article
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Background Both selenium (Se) and probiotic Bacillus regulate the metabolism to help defense clod stress and improve the meat quality in breeding chicks. The purpose of this study was to evaluate the effect of supplemental Se and Bacillus in the form of Se-enriched Bacillus (SECB) on the growth performance, lipid parameters, breast Se and antibiotic levels, and breast meat quality of chicken in winter cold stress. Methods Five hundred 1-d-old chickens were divided into five groups randomly: Control, inorganic Se, compound Bacillus, SECB, and antibiotic. The feed duration was 56 d. Results After 28 d of treatment, chicks feed SECB or compound Bacillus had higher body weights than the control, and after 56 d, chicks given either SECB or compound Bacillus had higher body weights than the control chicks or those given inorganic Se. Adding SECB to feed significantly increased the lightness, redness, and yellowness of breast meat, improved the water-holding capacity, and reduced the shear force and cooking loss. The concentration of Se in the breast muscle very significantly increased after SECB and inorganic Se supplementation, which was opposite to the concentration of flavomycin in antibiotic supplemented chicks. The antioxidative status of plasma and breast meat was significantly improved with added compound Bacillus and SECB: the total antioxidant capacity, total superoxide dismutase, and glutathione peroxidase ability in the breast muscle significantly improved, and the malondialdehyde concentration in plasma decreased. The levels of total cholesterol plasma triglyceride and very-low-density lipoprotein cholesterol in the plasma and breast muscle was decreased compared to that of the control, while the plasma high-density lipoprotein cholesterol concentration increased. Conclusions In conclusion, SECB supplementation promoted the body growth, antioxidative status, and Se concentrations in the plasma and breast meat, and also improved the breast meat quality.
... Alcune formulazioni di probiotici contenenti B. coagulans sono commercialmente disponibili, compreso B. coagulans GBI-30, 6086 (commercializzato come Ganeden BC30, e di seguito denominato BC30). Quest'ultimo si caratterizza per la capacità di sopravvivere ai processi produttivi e di tollerare le alte temperature, mantenendo le sue proprietà probiotiche anche durante la conservazione (Bader et al., 2012). Questo ceppo sporigeno ha ottenuto lo status di "generalmente riconosciuto come sicuro" (GRAS) nel 2012 dalla Food and Drug Administration (FDA), a seguito dei risultati di studi tossicologici (Endres et al., 2011). ...
Article
A new functional pasta using a durum wheat flour rich in polyphenols, with added barley β-glucan and Bacillus coagulans GBI-30, 6086 was developed. Chemical, nutritional and microbiological analyses were carried out to perform a comprehensive evaluation of the final product quality. The settled debranning process of durum wheat produced a flour rich in proteins and phenolic acids (18.18 %s.s. and 392.1 µg/g respectively) that had a positive influence on pasta cooking performances as well as on its nutritional properties. The probiotic strain remained viable during the pasta-making and cooking processes, and its amount in cooked pasta (about 9.0 log CFU/100 g) would be considered sufficient to exert beneficial effects on consumer. The glycemic index of pasta measured in healthy volunteers was 59.7 and the functional ingredients addition did not affect the postprandial glycemic response of pasta in healthy volunteers.
... The use of bacterial spore formers, Bacillus spp. in particular, as probiotics could provide practical advantages for both humans and animals [5]. The ability to form spores endows probiotics with higher resistance to technological stresses during production and storage processes and also higher resistance to gastric (pH, digestive enzymes) and intestinal environmental conditions [6]. Indeed, a number of products containing spores of Bacillus spp. ...
Article
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Aim: To investigate the effect of oral administration of two Bacillus strains on fecal coliforms, Lactobacillus and Bacillus spp. in rat animal model. Materials and Methods: An in vivo experiment was conducted for 49-day period on 36 adult male albino Wister rats divided equally into to four groups. After 7-day adaptation period, one group (T1) was fed on sterile skim milk along with basal diet for the next 28 days. Second (T2) and (T3) groups received spore biomass of Bacillus coagulans B37 and Bacillus pumilus B9, respectively, suspended in sterilized skim milk at 8-9 log colony-forming units/ml plus basal diet for 28 days, while control group (T4) was supplied with clean water along with basal diet. There was a 14-day post-treatment period. A total of 288 fecal samples (8 fecal collections per rat) were collected at every 7-day interval starting from 0 to 49 days and subjected to the enumeration of the counts of coliforms and lactobacilli and Bacillus spores using respective agar media. In vitro acid and bile tolerance tests on both the strains were performed. Results: The rats those (T2 and T3) received either B. coagulans B37 or B. pumilus B9 spore along with non-fermented skim milk showed decrease (p
... When spores of various Bacillus and Clostridium species survive such processes and germinate under conditions that support growth, these organisms can cause food spoilage, leading to significant economic losses (Rosenkvist and Hansen, 1995;Scheldeman et al., 2005Scheldeman et al., , 2006 and can in some cases be the cause of foodborne diseases (Lund, 1990). In contrast, commercially produced Bacillus spore products are increasingly applied as natural insecticides for crop protection or as probiotics (Jensen et al., 2003;Cutting, 2011;Bader et al., 2012). ...
Article
Spore germination shows a large inter-strain variability. Spores of certain Bacillus subtilis strains, including isolates from spoiled food products, exhibit different germination behavior from spores of the well-studied model organism Bacillus subtilis 168, often for unknown reasons. In this study, we analyzed spore germination efficiencies and kinetics of seventeen B. subtilis strains with previously sequenced genomes. A subsequent gene-trait matching analysis revealed a correlation between a slow germination phenotype and the presence of a mobile genetic element, i.e. a Tn1546-like transposon. A detailed investigation of the transposon elements showed an essential role of a specific operon (spoVA(2mob) ) in inhibiting spore germination with nutrients and with the cationic surfactant dodecylamine. Our results indicate that this operon negatively influences release of Ca-DPA by the SpoVA channel and may additionally alter earlier germination events, potentially by affecting proteins in the spore inner membrane. The spoVA(2mob) operon is an important factor that contributes to inter-strain differences in spore germination. Screening for its genomic presence can be applied for identification of spores that exhibit specific properties that impede spore eradication by industrial processes. This article is protected by copyright. All rights reserved.
... Бактерії роду Bacilus продукують велику кількість проти мікробних речовин [34]. Вони включають бактеріоцини та бактеріоциноподібні ингибуючі речовини (Subtilin, Coagu lin), а також поліпептидні антибіотики (Surfactin, Iturin, Bacilysin) [28]. Крім того, бактерії роду Bacillus у процесі своєї життєдіяльності синтезують різні за природою біоло гічно активні речовини та активізують власний синтез анти мікробних пептидів епітелієм ШКТ [33]. ...
Article
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The data of literature about spore-forming probiotics, as well as their usage in the pathology of the gastrointestinal tract in children were sumerised. Analysis of the data allows us to recommend the preparation «Biosporin-Biopharma» for preventive measures and treatment of gastrointestinal tract diseases at all levels of children health care, including infants.
... Bacterial spores are much more resistant to the listed conditions in the stomach and small intestine than living probiotic microorganisms like lactic acid bacteria. Hence they can reach the intestine in higher concentrations and might be more effective as probiotics [17]. Bacillus subtilis and Bacillus licheniformis had been used in human and livestock decades for regulation of innate and adaptive immune responses [18][19][20]. ...
Article
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Although breeding of F4 receptor − negative (F4R−) pigs may prevent post-weaning diarrhea, the underlying immunity is poorly understood. Here, various doses of a Bacillus licheniformis and Bacillus subtilis mixture (BLS-mix) were orally administered to F4ab/acR− pigs for 1 week before F4 (K88) − positive ETEC/VTEC/EPEC challenge. Administration of BLS-mix increased the percentage of Foxp3−IL-10+ T cells but not of Foxp3+IL-10+ regulatory T (Treg) cells among peripheral blood CD4+ T cells. A low dose of BLS-mix feeding resulted in increased the expression of IL-6, TNF-α, IL-10, and the transcription factors Foxp3 and T-bet mRNAs in the jejunum. Administration of either a low or high dose BLS-mix also led to an increase in the percentage of CD4+Foxp3+ Treg cells among intraepithelial lymphocytes and CD4+IL-10+ T cells in the small intestinal Peyer’s patches and the lamina propria of F4ab/acR− pigs following F4+ ETEC/VTEC/EPEC challenge. The increased number of IL-10–producing CD4+ T cells was attributed to an increase in the proportion of Foxp3−IL-10+ Treg cells rather than Foxp3+IL-10+ Treg cells. Our data indicate that oral administration of BLS-mix to newly weaned F4ab/acR− pigs ameliorates enteritis in an F4+ ETEC/VTEC/EPEC model; however, induction of IL-10–producing Foxp3− Treg cells by BLS-mix administration cannot account for the protection of newly weaned F4ab/acR− pigs from F4+ ETEC/VTEC/EPEC infection, and that excessive generation of CD4+IL-10+ T cells following consumption of BLS-mix during episodes of intestinal inflammation that is caused by enteric pathogens might prohibit clearance of the pathogen. Select probiotic mixtures may allow for tailoring strategies to prevent infectious diseases. Electronic supplementary material The online version of this article (doi:10.1186/s13567-015-0223-y) contains supplementary material, which is available to authorized users.
... The majority of microorganisms used as probiotics belong to the lactic acid bacteria (Burgain et al. 2014). However, sporeforming health promoting bacteria are broadly used in agricultural practice (Quigley et al. 2013) and are on the rise in various human applications, with the Clostridium and Bacillus genera being among them (Cutting 2011;Bader et al. 2012). ...
Article
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Unlabelled: DNA protective and antioxidant activity of Bacillus amyloliquefaciens B-1895 and Bacillus subtilis KATMIRA1933 were evaluated by Escherichia coli-based Lux biosensors. Two biosensor strains of E. coli, MG1655 (pColD-lux) and MG1655 (pSoxS-lux), which react on DNA damage and superoxide-anion radical activity, were used. SOS-response and Sox-response were stimulated by addition of dioxidine (2,3-Quinoxalinedimethanol,1,4-dioxide) and paraquat (N,N'-dimethyl-4,4'-bipyridinium dichloride) respectively. Preparations of both Bacillus fermentates demonstrated DNA protective and antioxidant (superoxide scavenging) activity (up to 60·19%). The strain К1933 is, in general, characterized by higher DNA protective activity (28·85%), with parameters of antioxidant activity of both bacilli strains being statistically not significantly different. Sporogenous potential probiotic micro-organisms with antioxidant and DNA protective activities can become an effective tool for compensation of various negative oxidative stress processes in humans. Significance and impact of the study: In humans, oxidative stress is a cause or an important component of many serious diseases, as well as being one of the age influencing factors. Environmental stresses lead to the increase in levels of reactive oxygen species (ROS). Oxidative DNA damage is a side effect of nonspecific inflammation. These human health challenging factors trigger the search for health-promoting bacteria capable of production of antioxidants and DNA-protectors. In this study, two Bacillus strains of interest were shown to produce noticeable DNA protective and antioxidant activities.
... For their spore-forming nature, these latter organisms possess improved viability and stability compared to other probiotic bacterial strains, e.g. the ability to withstand high temperature processes, such as baking and boiling, thus representing an ideal choice for the development of functional cereal-based products. Indeed, the endospores of the genus Bacillus are a very stable duration form and show high stability during the processing and storage of the food product (Bader et al., 2012). Further, in many food products, germination of the spores does not occur and thus the product quality of the food is not affected because of their inactive metabolism. ...
Article
Abstract A new functional pasta using a durum wheat flour rich in polyphenols, with added barley β-glucan and Bacillus coagulans GBI-30,6086 was developed. Chemical, nutritional and microbiological analyses were carried out to perform a comprehensive evaluation of the final product quality. The settled debranning process of durum wheat produced a flour rich in proteins and phenolic acids (18.18 % dm and 392.1 µg/g respectively). These raw material characteristics had a positive influence on pasta cooking performances as well as on its nutritional profile due to the high content of bound ferulic acid content (94% of total phenolic acids). The probiotic strain remained viable during the pasta-making and cooking processes, and its amount in cooked pasta (about 9.0 log CFU/100 g) would be considered sufficient to exert beneficial effects on consumer. The glycemic index of pasta measured in healthy volunteers was 59.7 and the functional ingredients addition did not affect the postprandial glycemic response of pasta in healthy volunteers. The information provided here suggests that the new developed functional pasta could represent a high-quality product for its suitable nutritional appeal and potential beneficial properties.
... Although the use of spores of some Bacillus or Clostridium spp. may appear controversial from a safety point of view, their advantages like heat stability, resistance to low pH of stomach and to other deleterious conditions like bile secretions in the intestinal environment, long term storage without refrigeration, and no need for encapsulation, has increased research interest in them compared with the more vulnerable vegetative cells (Bader et al., 2012;Foligne et al., 2012). While a substantial number of microbial species have been reported to exhibit potential probiotic properties (Holzapfel et al., 2001), only the most documented and robust strains make it to the market (Foligne et al., 2013). ...
Chapter
In recent times, increasing evidence of benefits of probiotics, for health restoration coupled with consumer's inclination towards safe, natural, and cost effective substitutes for drugs, has generated immense interest in exploring their therapeutic potential. Administering probiotics as such not only compromises their viability but also fails to guarantee their successful establishment in gut. In recent times emphasis has been made to take a pharmaceutical approach to probiotics which will not only maintain their viability during storage but will also assist in their successful delivery to and establishment at the site of action in a viable form. Further to above, supplementation of probiotics with prebiot-ics is also gaining predominance in the recent past. Numerous reports indicate that co-encapsulating prebiotics with probiot-ics, improves viability of the latter, apart from the independent growth promoting effect of the former on the inherent healthy gut microbiota of the host. The current chapter aims to analyse probiotics as pharmaceuticals , their encapsulation techniques, existing pharmaceutical preparations, their regulatory status in some countries, and future research needs. The current status of prebiotics in health care is also highlighted.
... Natto) have been used as probiotics for human consumption (13,14). Considering their spore-forming nature, probiotic Bacillus strains possess improved viability and survivability as well as probiotic properties during the harsh manufacturing processes (like cooking), the storage of the product, and finally, passage through the gastrointestinal tract as compared to other probiotic bacterial strains (6,(15)(16)(17)(18). In other words, sporeforming probiotic bacteria could be considered as a valuable solution for overcoming the limitations related to the stability of probiotic strains of Lactobacillus and Bifidobacterium during processing and storage of foods (6,7,11,12,19). ...
Article
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Background and Objectives: The use of Bacillus probiotics is an interesting way for manufacturing healthier meat and meat products as functional foods and also a valuable solution for overcoming the constraints related to the stability of probiotic strains of Lactobacillus and Bifidobacterium during processing and storage of foods. This work is an attempt to investigate the influence of common household cooking methods on the viability of Bacillus coagulans and Bacillus subtilis as probiotic sporeformers inoculated into cooked sausages. Materials and Methods: After preparation of the final suspensions of Bacillus coagulans ATCC 31284 and Bacillus subtilis var. Natto ATCC 15245 spores, they were supplemented in the sausage samples with different meat types and percentages. The inoculated cooked sausages were then submitted to common household cooking methods, i.e. boiling, microwaving, and deep fat frying. Enumeration of the studied spores was carried out on trypticase soy agar after exposure to a heat shock at 68°C for 20 min. One-way analysis of variance (ANOVA) of SPSS-20 was used for statistical analysis of the data. A Taguchi L 9 experimental design comprising 3 variables and 3 levels was performed using MINITAB-17 to determine main effects of the parameters of household cooking process. Results: The results obtained in this study showed that boiling, followed by microwaving and deep fat frying, had the greatest viability and recovery of the studied spores supplemented in the cooked sausages, respectively (P < 0.05). Furthermore, results of the Taguchi experimental method revealed that the type of Bacillus probiotics, followed by the formulation of cooked sausages and the method of household cooking of cooked sausages had main effects on the concentrations of the spores examined, respectively. Conclusions: The information provided here suggests that boiling as a common household cooking method retained maximum viability and recovery of the studied Bacillus probiotics supplemented in the cooked sausage samples. Besides, the type of spores was the principle parameter, which had a critical effect on the response (i.e. concentrations of the studied Bacillus probiotics).
... Bacillus spp., are Gram-positive, rod-shaped, and spore-forming bacteria. They are suitable to be consumed by living organisms due to their capability to withstand highly acidic conditions (Bader et al., 2012). ...
Article
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The presence of heavy metals in aquaculture is a major concern due to possible toxicity effects to the organisms. Bioaccumulation with bacteria is an effective and economical way to remove heavy metals from the water. The objectives of this research were to measure the growth rate of probiotics (Bacillus sp. BpChIAY [BpChIAY] and Bacillus thruingiensis, [Bt]) under different concentrations of selected heavy metals, and to determine the ability of the probiotics to bioaccumulate selected metals. Bacterial strains were grown in nutrient broth with the addition of heavy metals (Cu, Cr, Cd, Zn, Ni) at 37oC to determine the growth under exposure to heavy metals. The bioaccumulation experiment was conducted by exposing the strains' pellets to heavy metals solutions. The concentrations of heavy metals were measured using Atomic Absorption Spectroscopy (AAS). The bacterial growth percentages when grown under 2 ppm Zn, Ni and Cd for BpChIAY were 128%, 103% and 67%, and for BT, the growth rates were 97.7%, 98.8% and 36.2%, respectively. The findings showed the order of the toxicity in the order from the most toxic: Cu>Cr>Cd>Zn>Ni. Both strains were able to bioaccumulate the heavy metals, and BpChIAY was a better metal absorber than BT at the rate of 0.0539 mg/g for Zn, 0.0781 mg/g for Ni and 0.0256 mg/g for Cd.
... 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. ...
Book
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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.
... 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). ...
Article
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.
... 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. ...
... Bacillus coagulans has been found to be an effective biotherapeutic agent in human and animal health promotion programmes (Bader et al. 2012;Aminlari et al. 2018;Haldar and Gandhi 2019). However, inadequate information is available for vacuum-dried milk powder with Bacillus probiotics. ...
Article
The present study explored the possibilities of using Bacillus coagulans as a probiotic culture in vacuum‐dried milk powder. The operational drying temperature at 63 ± 2 °C under 0.7 kg/cm2 pressure emerged as the best temperature at which to prepare dried milk powder within 4.5 h with a mean (±SEM) Bacillus coagulans B37 spore count of 8.78 ± 0.03 log cfu/g and moisture content of 4.82 ± 0.12%. The total spore counts in vacuum‐dried milk powder did not change (P > 0.10) at storage temperatures of 7 °C, 37 °C or 45 °C over a three‐month period.
... Most commercial strains of probiotics are inherently thermolabile and must, therefore, be protected from excess heat. Nevertheless, some exceptions exist, such as Bacillus probiotics, which are also known as soil-based organisms or "SBO" (Bader, Albin, & Stahl, 2012). ...
Article
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The potential health benefits of probiotics may not be realized because of the substantial reduction in their viability during food storage and gastrointestinal transit. Microencapsulation can be used to enhance the resistance of probiotics to unfavorable conditions. A range of oral delivery systems has been developed to increase the level of probiotics reaching the colon including embedding and coating systems. This review introduces emerging strategies for the microencapsulation of probiotics and highlights the key mechanisms of their stress–tolerance properties. Recent in vitro and in vivo models for evaluation of the efficiency of probiotic delivery systems are also reviewed. Encapsulation technologies are required to maintain the viability of probiotics during storage and within the human gut so as to increase their ability to colonize the colon. These technologies work by protecting the probiotics from harsh environmental conditions, as well as increasing their mucoadhesive properties. Typically, the probiotics are either embedded inside or coated with food‐grade materials such as biopolymers or lipids. In some cases, additional components may be coencapsulated to enhance their viability such as nutrients or protective agents. The importance of having suitable in vitro and in vivo models to evaluate the efficiency of probiotic delivery systems is also emphasized.
... 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. ...
Article
Full-text available
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). ...
Article
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. ...
Article
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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.
... Bacillus sp. is capable of spore-forming. Compared with the lactic acid bacteria, spore-forming bacteria have several advantages, such as a higher survival rate on the gastrointestinal tract and stability during processing and storage [24]. erefore, acid-producing bacteria (B. ...
Article
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In this study, the application of an autochthonous microorganism as probiotic on catfish (Clarias sp.) was scarcely reported. This study aimed to obtain probiotic candidates from the digestive tract (intestinal and gastric) of catfish. A total of nine isolates were successfully isolated from the catfish. Almost all bacterial colonies were morphologically round, had flat edges, were yellow, and produced clear zones as a sign of producing acid during culture. The analysis showed that the three isolates had the best activity in inhibiting fish pathogen isolates. Furthermore, molecular analysis revealed that those three isolates were Bacillus velezensis UB-C1, Bacillus amyloliquifaciens UB-C5, and Bacillus cereus UB-C8. Interestingly, those three bacteria were non-lactic acid bacteria.
... 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]. ...
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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.
... There has been recent interest in utilizing Clostridia as probiotics and there is potential for considering these organisms as probiotics, but they could be difficult to produce since they are anaerobes. Clostridia are already on the market as probiotics for human gastrointestinal applications, so this is an opportunity for new investigations into potential use of clostridia as probiotics during foodanimal production [82]. Finally, the in ovo methodology can be adopted to supply the chicken embryo with additional nutrients prior to hatching which will continue to be utilized by the chick post-hatch during the fasting period. ...
... Unlike non-sporeforming probiotics (e.g. strains of lactobacilli and bifidobacteria), sporeformers have a better survival rate when passing through the highly acidic environment of stomach and small intestine, and are more stable during product manufacturing and storage [12,18]. ...
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Probiotic preparations are used in medical treatment and in agricultural practice. They modulate numerous activities in eukaryotic hosts, such as: inhibition of pathogenic microbiota; stimulation of immunological responses; and production of antioxidants, anti-mutagens, and DNA protectors. Also, probiotic bacteria are used as a preventive measure to prevent bacterial diseases of the gastrointestinal tract. Solid-phase fermentation is reported as being used in the production of probiotic formulations where a solid substratum, such as soy and oil meal, is utilized for the growth of beneficial microorganisms. However, there are insufficient reports in the literature related to methodological approaches enabling evaluation of the final products of solid-phase fermentation. We suggest a novel method enabling evaluation of probiotic solid-state fermentation dry powders and observation of their morphology, ultrastructure, and elucidation of the quantitative distribution of probiotic microorganisms in solid substrates using electron microscopy. •The method is intended for ultrastructure microphotography of dry substances - for example, ultrastructure of solid-phase fermentation products.•The method allows preserving the ultrastructure of substrates that are damaged when soaking.•The method does not require additional equipment and reagents and can be used in all laboratories using electron microscopy.
... 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] . ...
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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.
... 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]. ...
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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.
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The use of food-grade organisms as recombinant vaccine expression hosts and delivery vehicles has been explored during the past 25 years, opening new avenues for vaccinology. Considering that oral immunization is a beneficial approach in terms of costs, patient comfort, and protection of mucosal tissues, the use of food-grade organisms can lead to highly advantageous vaccines in terms of costs, easy administration, and safety. The organisms currently used for this purpose are bacteria (Lactobacillus and Bacillus), yeasts, algae, plants, and insect species. Herein, a comparative and updated scenario on the production of oral vaccines in food-grade organisms is provided and placed in perspective. The status of clinical evaluations and the adoption of this technology by the industry are highlighted.
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Objective: The purpose of this randomized double-blind placebo controlled clinical trial was to evaluate the effects of a commercial spore-based probiotic supplement consisting of 5 different spore forming bacilli (Bacillus indicus HU36, Bacillus subtilis HU58, Bacillus coagulans SC-208, Bacillus licheniformis, and Bacillus clausii SC-109) on reducing the triglyceride levels (TG) in patients with mild to moderate hypertriglyceridemia (HT). Study design: A randomized, double-blind, placebo-controlled study with eighty participants with non-fasting triglyceride levels greater than 150 mg/dL. Methods: Eighty participants with non-fasting triglyceride levels greater than 150 mg/dL were randomized to receive oral probiotic supplement consisting of two capsules containing 5 different spore forming bacilli once daily in the morning or a placebo (rice flour). Their non-fasting triglyceride levels were measured again at six weeks and at twelve weeks. Results: Compared to the placebo group, participants in the probiotic supplement group had significant lowering of their triglyceride levels after 90 days. Conclusion: Mild to moderately elevated triglyceride levels can be lowered in patients with mild to moderate HT by a probiotic supplement consisting of five different spore forming bacilli.
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In inflammatory bowel disease (IBD), experimental models have proven to be important tools for evaluating potential therapeutic agents and for investigating the mechanisms of pathogenesis. Oxidative stress and the immune response have been associated with acetic acid (AA)-induced ulcerative colitis (UC). Our study aimed to evaluate, for the first time, the ability of a spore-based probiotic and an amino acid and immunoglobulin supplement in reducing tissue damage and inflammatory responses in an experimental animal model of UC. Forty-two Wistar rats were divided into six groups, receiving 1% carboxymethylcellulose, 4% AA, MegaSporeBiotic™ (MSB; 1 × 10 9 colony forming units/day) and MegaMucosa™ (MM; 70 mg/100 g/day). Pretreatment with MSB or MM alone and in combination significantly lowered inflammation and reduced damage to the colonic mucosa. Pretreatment with these agents resulted in levels of proinflammatory cytokines, vascular tight junction proteins, and measures of oxidative stress similar to those reported for methylprednisolone, one of the first-line therapies for moderate to severe activity of UC. The protection was further confirmed by histologic analysis of the colon tissue. In conclusion, pretreatment with probiotic spore-forming Bacillus strains and a supplement of amino acids in combination with immunoglobulins exhibited anti-inflammatory and antioxidant effects in an AA-induced rat model of UC.
Chapter
Members of the Gram‐positive genera Bacillus and Clostridium and some closely related genera respond to slowed growth or starvation by initiating the process of sporulation, and the resultant spores can cause practical problems in food microbiology as well as human disease. The molecular biology of sporulation and spore resistance and germination in Bacillus subtilis has been extensively studied for many years, and there is detailed knowledge of these processes and many of the regulatory mechanisms involved. With the recent availability of many genome sequences as well as methods for genetic manipulation of clostridia of industrial and medical importance, a detailed molecular understanding of sporulation and spore germination in clostridia is also being developed. This chapter describes the fundamental bases of sporulation, spore germination, and spore resistance and the problems that spores present to the food industry.
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Salmonella infections have become a major health concern in recent decades. This pathogen has evolved to become resistant to antibiotics, which has caused problems in its treatment. As such, finding a novel preventive method is important in the treatment and management of this infection. In recent years, uses of probiotics, especially spore-former genera such as Bacillus spp. has become increasingly popular. In this study spores of two probiotic bacteria, Bacillus subtilis and Bacillus coagulans were fed to rats for three weeks through their daily water intake after which Salmonella Typhimurium was gavaged to the rats. On days 1, 3, 5 and 7 after gavaging, the number of Salmonella was counted in liver, spleen, mesenteric lymph nodes, feces and content of ileum and cecum. Hematological and biochemical parameters, inflammatory mediators, total antioxidant capacity and malondialdehyde were also measured. The results showed that B. subtilis and B. coagulans caused delation in infiltration of Salmonella into the lymph nodes, spleen and liver, reduction of the inflammatory mediators, and decreases in oxidative stress, hematological and biochemical changes. The overall count of Salmonella in the above mentioned parameters has also decreased and a faster return to normal base were also witnessed. The results showed that the use of B. subtilis and B. coagulans can potentially help boost the body’s immune system, to combat the effects of exposure to the Salmonella pathogen.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.