Probiotic Spores Research Articles

Enhancement of the Anti-inflammatory Effect of Bromelain by Its Immobilization on Probiotic Spore of Bacillus cereus.

The therapeutic application of bromelain is limited due to its sensitivity to operating conditions such as high acidity, gastric proteases in the stomach juice, chemicals, organic solvents and elevated temperature. We hypothesized that bromelain immobilized on probiotic bacterial spores would show enhanced therapeutic activity through possible synergistic or additive effects. In this study, the oedema inhibition potential of bromelain immobilized on probiotic Bacillus spores was compared to the free enzyme using the carrageenan paw oedema model with Wistar rats. In batch A rats (carrageenan-induced inflammation 30min after receiving oral treatments), group 7 rats treated with a lower dose of spore-immobilized bromelain suspension showed the highest oedema inhibition, 89.20±15.30%, while group 4 treated with a lower dose of free bromelain had oedema inhibition of 60.25±13.00%. For batch B rats (carrageenan-induced inflammation after receiving oral treatment for three days), group 7 rats treated with a lower dose of spore-immobilized bromelain suspension showed higher inhibition percentage (81.94±8.86) than group 4 treated with a lower dose of free bromelain (78.45±4.46) after 24h. Our results showed that used alone, the enzyme and the spores produced oedema inhibition and improved the motility of the rats. The spore-immobilized bromelain formulation performed approximately 0.9-fold better than the free bromelain and the free spores at the lower evaluated dose.

Prebiotics-Encapsulated Probiotic Spores Regulate Gut Microbiota and Suppress Colon Cancer.

While microbial-based therapy has been considered as an effective strategy for treating diseases such as colon cancer, its safety remains the biggest challenge. Here, probiotics and prebiotics, which possess ideal biocompatibility and are extensively used as additives in food and pharmaceutical products, are combined to construct a safe microbiota-modulating material. Through the host-guest chemistry between commercial Clostridium butyricum and chemically modified prebiotic dextran, prebiotics-encapsulated probiotic spores (spores-dex) are prepared. It is found that spores-dex can specifically enrich in colon cancers after oral administration. In the lesion, dextran is fermented by C. butyricum, and thereby produces anti-cancer short-chain fatty acids (SCFAs). Additionally, spores-dex regulate the gut microbiota, augment the abundance of SCFA-producing bacteria (e.g., Eubacterium and Roseburia), and markedly increase the overall richness of microbiota. In subcutaneous and orthotopic tumor models, drug-loaded spores-dex inhibit tumor growth up to 89% and 65%, respectively. Importantly, no obvious adverse effect is found. The work sheds light on the possibility of using a highly safe strategy to regulate gut microbiota, and provides a promising avenue for treating various gastrointestinal diseases.

Bacillus subtilis spore vaccines displaying protective antigen induce functional antibodies and protective potency

BackgroundBacillus anthracis is the causative agent of anthrax, a disease of both humans and various animal species, and can be used as a bioterror agent. Effective vaccines are available, but those could benefit from improvements, including increasing the immunity duration, reducing the shot frequency and adverse reactions. In addition, more sophisticated antigen delivery and potentiation systems are urgently required.The protective antigen (PA), one of three major virulence factors associated with anthrax was displayed on the surface of Bacillus subtilis spores, which is a vaccine production host and delivery vector with several advantages such as a low production cost, straightforward administration as it is safe for human consumption and the particulate adjuvanticity. Mice were immunized orally (PO), intranasally (IN), sublingually (SL) or intraperitoneally (IP) with the PA displaying probiotic spore vaccine. Clinical observation, serological analysis and challenge experiment were conducted to investigate the safety and efficacy of the vaccine.ResultsA/J mice immunized with the PA spore vaccine via PO, IN, SL, and IP were observed to have increased levels of active antibody titer, isotype profiles and toxin neutralizing antibody in sera, and IgA in saliva. The immunized mice were demonstrated to raise protective immunity against the challenge with lethal B. anthracis spores.ConclusionsIn this study, we developed a B. subtilis spore vaccine that displays the PA on its surface and showed that the PA-displaying spore vaccine was able to confer active immunity to a murine model based on the results of antibody isotype titration, mucosal antibody identification, and a lethal challenge experiment.

Application of spore-forming probiotic Bacillus in the production of Nabat - A new functional sweetener

The basis of the present study was developing a novel type of functional sweet product with the additive probiotic spore forming bacteria that do not require cold chain or any specific storage condition. In this study, rock candy containing probiotic bacteria was produced in dry ambient condition. Spore-forming bacilli were selected as probiotics that are resistant to thermal processes and osmotic conditions. Rock candies were coated with an edible coating containing carboxymethyl cellulose, sucrose, glucose syrup, fructose syrup and a probiotic strain. After coating process, the parameters of turbidity (created by water dissolution of coated Nabat); effect of drying temperature on bacterial viability; moisture content, water activity, ash content of product; as well as product storage time were evaluated. Under thermal stress during both dissolution in hot liquid and drying, the survivability of the spores used in the coated rock candies did not fall significantly. Viability evaluation of the probiotic bacilli in the product after six months of storage at ambient temperature revealed 90.39 and 89.07% viability of Bacillus coagulans and Bacillus subtilis spores, respectively. Our results showed that spore-forming probiotics embedded in rock candy are highly stable during storage at ambient temperature and dry condition.

Synbiotic Supplementation Containing Whole Plant Sugar Cane Fibre and Probiotic Spores Potentiates Protective Synergistic Effects in Mouse Model of IBD.

Inflammatory bowel diseases (IBD) are a chronic inflammatory disorders with increasing global incidence. Synbiotic, which is a two-point approach carrying probiotic and prebiotic components in mitigating inflammation in IBD, is thought to be a pragmatic approach owing to the synergistic outcomes. In this study, the impacts of dietary supplementation with probiotic Bacillus coagulans MTCC5856 spores (B. coagulans) and prebiotic whole plant sugar cane fibre (PSCF) was assessed using a murine model of IBD. Eight-week-old C57BL/6 mice were fed a normal chow diet supplemented with either B. coagulans, PSCF or its synbiotic combination. After seven days of supplementation, colitis was induced with dextran sulfate sodium (DSS) in drinking water for seven days during the continuation of the supplemented diets. Synbiotic supplementation ameliorated disease activity index and histological score (−72%, 7.38, respectively), more effectively than either B. coagulans (−47%, 10.1) and PSCF (−53%, 13.0) alone. Synbiotic supplementation also significantly (p < 0.0001) prevented the expression of tight junction proteins and modulated the altered serum IL-1β (−40%), IL-10 (+26%), and C-reactive protein (CRP) (−39%) levels. Synbiotic supplementations also raised the short-chain fatty acids (SCFA) profile more extensively compared to the unsupplemented DSS-control. The synbiotic health outcome effect of the probiotic and prebiotic combinations may be associated with a synergistic direct immune-regulating efficacy of the components, their ability to protect epithelial integrity, stimulation of probiotic spores by the prebiotic fibre, and/or with stimulation of greater levels of fermentation of fibres releasing SCFAs that mediate the reduction in colonic inflammation. Our model findings suggest synbiotic supplementation should be tested in clinical trials.

Bacillus clausii UBBC-07 reduces severity of diarrhoea in children under 5 years of age: a double blind placebo controlled study.

Acute diarrhoea is one of the leading causes of mortality in infants and young children. Evidence suggests that probiotics can reduce diarrhoea duration. As the effects of probiotics are strain specific, the effect of Bacillus clausii UBBC-07, a safe probiotic strain in the treatment of acute diarrhoea in children was studied. The double blind, randomised, placebo-controlled, parallel group multicentric study was conducted at two outpatient facility sites in Lucknow, India. Children aged six months to five years suffering from acute diarrhoea, were randomly assigned to receive either probiotic (B. clausii UBBC-07) spore suspension or placebo suspension twice daily apart from oral rehydration solution (ORS). The duration of treatment was for five days with a follow -up until the 10th day. Outcomes evaluated were duration and frequency of diarrhoea, consistency of stool, fever and vomiting. The duration of diarrhoea was significantly shorter (P<0.05) in patients who received B. clausii suspension (75.66±13.23 h) than in placebo treated group (81.6±15.43 h). The average daily number of stools (frequency) was 8.67±3.42 at baseline in treatment group receiving B. clausii and 8.53±3.19 in placebo group. By day 4, there was a significant reduction (P<0.01) in frequency of stools in probiotic treated group (3.46±0.66) as compared to placebo group (4.57±1.59). Improvement in stool consistency was also observed in the probiotic treated group as compared to the placebo group. There was no effect on vomiting and duration of fever. B. clausii UBBC-07 significantly decreased the duration and frequency of diarrhoea as compared to placebo indicating effectiveness of strain in the treatment of acute diarrhoea in children and could be a safe alternative to antibiotics.

Probiotic Spore Formers Enhances Host Health

Probiotic Spore Formers Enhances Host Health

Antibiotic-associated disorders of prothrombin synthesis and their probiotic correction with B. Clausii in breastfeed infants

The aim of the investigation was to study the prothrombin test and PIVKA-II serum levels changes in infants aged 1-6 month after the broad-spectrum antibiotic treatment and determine the possibility of coagulation disorders correction with the help of probiotic Bacillus clausii strains OC , NR , T , SIN spores capable to produce vitamin K 2 . Materials and methods . The study involved 57 convalescents after an acute respiratory infection exclusively breastfed infants aged 1 to 6 months one week after the 7-day parenteral treatment with ceftriaxone. Among them 13 children with laboratory signs of blood hypocoagulation (INR >1.3) had received probiotic Bacillus clausii spores in a daily dose of 2 × 10 9 CFU for 10 days. Another 13 children who had not received a probiotic made up a comparison group. Laboratory data was compared with the results of 27 healthy children of the control group. Prothrombin test parameters as a prothrombin (PT), prothrombin time (PTT), international normalized ratio (INR) and protein induced by vitamin K absence or antagonist II (PIVKA-II) were determined using the hematologic analyzer COAGUCHEC XS and ELISA kit. Results . It was found that after antibacterial treatment in infants PT was decreased and reached only 50 % of the norm. At the same time, PTT and INR were increased up to 2-folds together with a significant PIVKA-II elevation, which is typical for vitamin K deficiency. The use of probiotic treatment within 10 days has contributed to normalization of prothrombin in 11 out of 13 observed infants versus 3 out of 13 in the comparison group (χ 2 = 7,58 corrected by Yates, P < 0.05). Conclusions . Cephalosporin administration for breastfed infants can lead them to latent vitamin K deficiency development, which is manifested by high levels of PIVKA-II and signs of hypocoagulation development revealed by prothrombin test. The Bacillus clausii probiotic spores prescription to infants after antibiotic therapy course for 10 days normalizes the plasma prothrombin concentration while in children of the comparison group it remains lower than normal.

Antagonistic activity of Bacillus subtilis strains isolated from various sources

&lt;p&gt;An important link in solving the problem of healthy food is the intensification of the livestock, poultry and fish farming, which is possible only in the adoption and rigorous implementation of the concept of rational feeding of animals. In the implementation of this concept required is the application of probiotic preparations. Currently, there is an increased interest in spore probiotics. In many ways, this can be explained by the fact that they use no vegetative forms of the bacilli and their spores. This property provides spore probiotics a number of advantages: they are not whimsical, easily could be selected, cultivated, and dried. Moreover, they are resistant to various factors and could remain viable during a long period. One of the most famous spore microorganisms, which are widely used in agriculture, is &lt;em&gt;Bacillus subtilis&lt;/em&gt;. Among the requirements imposed to probiotic microorganisms is mandatory – antagonistic activity to pathogenic and conditional-pathogenic microflora. The article presents the results of the analysis of antagonistic activity of collection strains of &lt;em&gt;B. subtilis&lt;/em&gt;, and strains isolated from commercial preparations. We studied the antagonistic activity on agar and liquid nutrient medias to trigger different antagonism mechanisms of &lt;em&gt;B. subtilis&lt;/em&gt;. On agar media, we applied three diffusion methods: perpendicular bands, agar blocks, agar wells. We also applied the method of co-incubating the test culture (&lt;em&gt;Escherichia coli&lt;/em&gt;) and the antagonist (or its supernatant) in the nutrient broth. Our results demonstrated that all our explored strains of &lt;em&gt;B. subtilis&lt;/em&gt; have antimicrobial activity against a wild strain of &lt;em&gt;E. coli&lt;/em&gt;, but to varying degrees. We identified strains of &lt;em&gt;B. subtilis&lt;/em&gt; with the highest antagonistic effect that can be recommended for inclusion in microbial preparations for agriculture.&lt;/p&gt;&lt;p&gt;&lt;em&gt;&lt;br /&gt;&lt;/em&gt;&lt;em&gt;&lt;/em&gt;&lt;/p&gt;

Effects of the dietary supplementation of mixed probiotic spores of Bacillus amyloliquefaciens 54A, and Bacillus pumilus 47B on growth, innate immunity and stress responses of striped catfish (Pangasianodon hypophthalmus)

The study used the mixed probiotics of Bacillus amyloliquefaciens 54A and B. pumilus 47B isolated from striped catfish (Pangasianodon hypophthalmus) intestine aiming to stimulate growth performance, innate immunity, stress tolerance of striped catfish. The average weight gain (AWG), specific growth rate (SGR), and feed conversion ratio (FCR) were analyzed after fish were fed the mixture of probiotics (B. amyloliquefaciens 54A and B. pumilus 47B) at concentrations of 1 × 108, 3 × 108, and 5 × 108 CFU g−1 feed for 90 days. Immunity parameters, survival rate of fish challenged with Edwardsiella ictaluri and ammonia tolerance were also investigated. The amounts of B. amyloliquefaciens and B. pumilus were counted and identified by specific primer pairs of Ba1-F/Ba1-R, and 16-F/Bpu-R to confirm the presence of probiotics in fish intestine. The AWG (476.6 ± 7.81 g fish−1) of fish fed probiotics at 5 × 108 CFU g−1 was significant higher than the control (390 ± 25.7 g fish−1) after 90 days of feeding, but there was no significant (P > 0.05) effect of probiotics on FCR and SGR. Fish fed diet containing probiotics at 5 × 108 CFU g−1 also expressed resistance to E. ictaluri infection and higher immune parameters such as phagocytic activity, respiratory bursts, and lysozyme activity than the control. Stress response with ammonia showed significantly lower mortality rate (25%, 20% and 27%) of fish fed probiotics at all three levels of 1, 3 and 5 × 108 CFU g−1 than the fish fed control diet (75%). The study also demonstrated that the probiotics survived in the intestine of striped catfish after 90 days of feeding. Therefore, the dietary supplementation of a mixture of B. amyloliquefaciens and B. pumilus at 5 × 108 CFU g−1 can be used to improve the health and growth rate of striped catfish.

Inhibition of Lipopolysaccharide-Induced Interleukin 8 in Human Adenocarcinoma Cell Line HT-29 by Spore Probiotics: B. coagulans and B. subtilis (natto).

Probiotics are used as a treatment for different intestinal disorders. They confer health benefits by different ways. This study was aimed to investigate immunomodulatory effect of Bacillus probiotic spores on the production of lipopolysaccharide (LPS)-induced interleukin 8 (IL-8) in HT-29 intestinal epithelial cells. Differentiated intestinal epithelial cell line was used as a model for the study of colonization of purified spores (Bacillus subtilis (natto) and B. coagulans) and their anti-inflammatory effects. MTT assay and trypan blue staining were used for the detection of optimal concentration of the purified spores and LPS. Pre-treatment assay was done by treatment of the cells with the purified spores for 2h, followed by challenges with LPS for 3 and 18h. Post-treatment assay was done by initial treatment of the cells with LPS for 18h, followed by the spores for 3 and 6h. Levels of IL-8 secretion and its mRNA expression were measured by ELISA and relative Q real-time PCR. Our results showed similar rates of adherence to intestinal epithelial cells by the spore probiotics, while displaying no cytotoxic effect. In the pre-treatment assay, a significant decrease in IL-8, at both protein and mRNA levels, was measured for B. coagulans spores after the addition of LPS, which was higher than those observed for Bacillus subtilis (natto) spores. In the post-treatment assay, while Bacillus subtilis (but not B. coagulans) diminished the LPS-stimulated IL-8 levels after 3h of incubation, the inhibitory effect was not constant. In conclusion, ability of Bacillus spore probiotics for adherence to intestinal epithelial cell and their anti-inflammatory effects, through interference with LPS/IL-8 signaling, was shown in this study. Further studies are needed to characterize responsible bacterial compounds associated with these effects.

Quality Characterization of Processed Cheese Inoculated by Bacillus coagulans During Cold Storage: Compositional and Sensorial Attributes and Probiotic Microorganism Viability

The effect of the addition of probiotic spores (107 and 108 colony-forming unit [cfu]/g) of Bacillus coagulans ATCC 7050 on the critical quality attributes of an Iranian processed cheese including chemical composition, pH, titratable acidity (TA), survival rate and sensory features was studied during 60 days of cold storage. No significant difference was found in the chemical composition by the increasing spore count. A low pH value and a high TA quantity in the cheese inoculated by Ba. coagulans spores in comparison with the control sample were observed (P < 0.05). Results showed that the viability of probiotic cells was mentioned more than 106 cfu/g during the cold storage. The color, odor and texture sensory scores were insignificantly affected by the addition of probiotic spores during storage. However, a significant difference was found for the flavor and overall acceptability among different samples as these sensory attributes decreased with an increase in spore count. Practical Applications Incorporation of probiotic bacteria has been successfully performed in whey cheese matrices. However, probiotic-processed cheese inoculated by Bacillus coagulans spores have not been before made according to the data found in the literature. The use of this microbial source in producing processed cheese led to the improved critical quality attributes during 60 days of storage at refrigerator temperature. This study demonstrated that the processed functional cheese can be a potential carrier for the delivery of probiotic cells.

Low cost medium for spore production of Bacillus KKU02 and KKU03 and the effects of the produced spores on growth of giant freshwater prawn (Macrobrachium rosenbergii de Man).

In order to extend the shelf life of 2 high potential Bacillus probiotic isolates which were Bacillus KKU02 and Bacillus KKU03, the spore forms of these 2 Bacillus isolates were studied for using as probiotic instead. The low cost medium for spore production of these 2 Bacillus isolates was examined in order to produce probiotic spores for feeding the shrimps. It was found that cassava at 100 g L(-1) and supplemented with 20.0 g L(-1) dextrose, 0.1 g L(-1) MgSO4 and 2.0 g L(-1) (NH4)2SO4 showed the highest spore concentration at about 1 x 10(8) CFU mL(-1). The effects of feeding these 2 Bacillus spores on the growth of giant freshwater prawns were further examined. The spores of Bacillus KKU02 and Bacillus KKU03 (-10(7) spore mL(-1)) in pure and mixed culture forms were mixed with commercial prawn feed (200 mL kg(-1)) to give six feed treatments. Body length and weight of the prawns in mixed spore culture tanks after rearing for 90 days (13.5 cm and 59.8 g, respectively) were significantly higher (p = 0.05) than other treatments. The treated prawns were further challenged with Aeromonas hydrophila for 7 days. The percentages of survival after the challenge in the prawns fed with the mixed spores (46.8%) were also found significantly higher (p = 0.05) than others groups, except the mixed live cell treatment (60%). These results indicated that the spores of Bacillus KKU02 and Bacillus KKU03 had a high potential for using as commercial probiotics.

Антагоністична активність пробіотиків із бацил до ізолятів бактерій ротової порожнини пацієнтів із пародонтитом

It is admitted that the normal human microflora plays an important role in supporting homeostasis, forming immune mechanisms and metabolism. Nowadays, there is a constant growth of different diseases due to microbiological imbalance in a human organism. Preparations containing “good bacteria” have been used for therapeutic purposes since ancient times. The mechanism of probiotics influence comprises their ability to compete for adhesion sites with the pathogens, to exhibit antagonistic activity and stimulate the immune system of a host. Most of probiotics commonly used are the spores of Bacillus. Initially the main focus of their use was the prevention of gastrointestinal disorders. So, the use of probiotics in dental practice is a poorly studied area. In recent years, probiotics have been investigated to provide the oral health. Therefore the study of using probiotics for correction of the oral microflora in people with inflammatory diseases of the periodontal tissues is promising. Our previous studies have shown changes in microbial community of an oral cavity in patients with periodontitis. In particular, the reducing number of obligate microorganisms and increasing number of pathogens was demonstrated. The paper describes the current data on the potential benefits and basic properties of the Bacillus spore probiotics, which demonstrate the viability and relevance in dental practice. The study tested antagonistic activity of commercial Bacillus probiotics "Biosporin" ("Biopharma", Ukraine), "Subalinum" ("Biopharma", Ukraine), "Normaflore" ("Sanofi-Aventis Zrt.", Hungary) and "Enterogermina" ("Sanofi-Synthelabo SpA", Italy) against clinical strains of microorganisms isolated from the oral cavity of patients with periodontitis. Thus, further studies on the role of spore probiotics in correction of the oral cavity microflora as a part of complex treatment of periodontitis should be carried out.

Time to Consider Clostridium Probiotics?

Time to Consider <i>Clostridium</i> Probiotics?

Investigation of the antagonistic activity of spore probiotics against Rhodococcus equi in experiments on mice

A method is developed for studying the antagonistic activity of the spore probiotics Sakhabactisubtil and Irilis in vivo against a virulent Rhodococcus equi. Intraperitoneal injection of sterile filtrates of these spore probiotics into mice that received the antibiotic Ampiox and were infected with a lethal dose of the virulent R. equi strain promotes the survival of 70–100% of the animals and reduced the number of rhodococci in internal organs by 6–9 times.

Biosynthesis of metal and oxide nanoparticles using Lactobacilli from yoghurt and probiotic spore tablets

Green, low-cost, and reproducible Lactobacillus-mediated biosynthesis of metal and oxide nanoparticles are reported. Silver and titanium dioxide nanoparticles are synthesized using Lactobacillus sp. procured from yoghurt and probiotic tablets. The synthesis is performed akin to room temperature in the laboratory ambience. X-ray and transmission electron microscopy analyses are performed to ascertain the formation of metallic and oxide nanoparticles. Individual nanoparticles having the dimensions of 10-25 nm (n-Ag) and 10-70 nm (n-TiO(2)) are found. The mechanism involved for the synthesis of metallic and oxide nanoparticles has also been discussed.

Oral Vaccine Delivery by Recombinant Spore Probiotics

Over the past few decades, advancements in molecular and cell biology have allowed scientists to identify a large number of new antigens from a variety of viral and bacterial pathogens. However, successful development of these antigens into effective vaccines strongly relies on delivery systems able to avoid the rapid loss of biological activity that often impairs antigen efficacy. Various delivery systems have been proposed as alternative vaccine vehicles, from live microorganisms to nanoparticles, and all of them have shown advantages but also drawbacks. The bacterial spore is a quiescent cell form that, as a vaccine vehicle, may conjugate some advantages of live microorganisms with those of synthetic nanoparticles and that has recently been proposed as a potentially powerful tool to deliver antigens to mucosal surfaces. Here we review the use of bacterial spores as a delivery system for mucosal immunizations. We will first analyze the nature of the interaction between wild type spores and the gut-associated lymphoid tissue and then address the immune responses that are induced by oral immunizations with recombinant spores displaying heterologous antigens.

Effectiveness of spore probiotics with respect to mycobacteria and nocardioform actinomycetes

The antagonistic activity of commercial bacillus-based probiotic preparations with respect to mycobacteria and nocardioform actinomycetes has been studied. A cell suspension or sterile culture filtrate of bacilli was plated on Loewenstein-Jensen medium and then indicator strains were inoculated.

Bacterial probiotics induce an immune response in the honey bee (Hymenoptera: Apidae).

To explore immune system activation in the honey bee, Apis mellifera L., larvae of four ages were exposed through feeding to spores of a natural pathogen, Paenibacillus larvae larvae, to cells of a diverse set of related nonpathogenic bacteria, and to bacterial coat components. These larvae were then assayed for RNA levels of genes encoding two antibacterial peptides, abaecin and defensin. Larvae exposed to either P. l. larvae or a mix of nonpathogenic bacteria showed high RNA levels for the abaecin gene relative to controls. First instars responded significantly to the presence of the nonpathogenic mix within 12 h after exposure, a time when they remain highly susceptible to bacterial invasion. This response was sustained for two successive instars, eventually becoming 21-fold higher in larvae exposed to probiotic spores versus control larvae. The mixture of nonpathogenic bacteria is therefore presented as a potential surrogate for assaying the immune responses of different honey bee lineages. It also is proposed that nonpathogenic bacteria can be used as a probiotic to enhance honey bee immunity, helping bee larvae, and other life stages, survive attacks from pathogens in the field.