Spore-forming Probiotics Research Articles

Effect of new probiotic of spore-forming Bacillus Subtilis, cultivated on a local agro-industrial waste, on rabbit productivity and meat quality

The aim of the research was to develop a low-cost, innovative and competitive methodology of production of spore-forming probiotics, demonstrating effectiveness as an alternative to antibiotics, by controlling and preventing gastrointestinal diseases in rabbits, improving their productivity and the quality of manufactured products, achieving economic well-being. A number of scientific and technological tasks were implemented in order to achieve this goal. Probiotic preparations and confirmation of their best effect was carried out in the rabbit enterprise “Organic Solution" LLC. As a result of the conducted 6 experiments, it was determined that the optimal dose of adding new spore-forming Bacillus subtilis probiotics to the feed of the rabbit is 0.04%. In the spore-forming probiotic material made from local agro-industrial raw materials, the spores retain their viability to an equal degree, so their storage in dry ground form is important from the point of view of practical use; The number of spores of the spore-forming probiotics of the fresh preparation mixed with granulated feed and stored for 6 months is unchanged and successfully maintains its activity. The research was carried out with the financial support of the Shota Rustaveli National Science Foundation (grant project CARYS-19-2268)

Employment of Spore-Forming Probiotics to Combat Persister Cells of Staphylococcus Epidermidis.

In this study, spore-forming probiotics were employed to eradicate Staphylococcus epidermidis biofilms and the presence and expression of genes involved in stress response was examined. Polymerase chain reaction (PCR) assay was used to detect rpoS, relA and mazF genes in S. epidermidis ATCC 12228. Biofilm production was investigated by microtiter plate (MTP) assay. 100X minimum inhibitory concentration (MIC) of gentamycin was used to induce persister cells in planktonic and biofilm bacterial cells. The expression of rpoS, relA, and mazF genes was assessed at different time intervals of 2, 8, and 24 h using real-time PCR assay. Then, dilutions of 1, 0.5, and 0.25 µg/ml of the supernatant of Bacillus coagulans culture was used to eradicate the persister cells and the number of colonies was determined. Persister cells of S. epidermidis were formed after 7 h in planktonic and 5 h in the biofilm structure after exposure to 50 µg/ml of gentamycin. The expression of mazF and rpoS in biofilm structure and the expression of rpoS and relA in persister cells were significantly higher compared to the control (p< 0.05). The number of persister cells showed a reduction of log 2.4 and log 0.8 after exposure to 1 and 0.5 µg/ml B. coagulans supernatant, respectively, but no reduction was observed at the concentration of 0.25 µg/ml. The results showed that the supernatant of probiotics containing their secretive metabolites can be used as a novel approach to combat persister cells.

Human in vivo assessment of the survival and germination of Heyndrickxia coagulans SNZ1969 spores delivered via gummy candies

Confectionary products hold promise as unconventional food carriers for probiotic microorganisms. This study explored the delivery of Heyndrickxia coagulans SNZ1969, a spore-forming probiotic, using gummy candies. In this study, we prepared gummy candies containing bacterial spores with a viable count that remained stable during a 24-month shelf-life period, meeting the label claim of at least one billion CFUs per serving (24 g). Then, we carried out an intervention trial involving 24 healthy adults who consumed one serving per day for two weeks followed by an additional two weeks of follow-up. Fecal samples were collected and analyzed with a protocol that allowed the viable counts of SNZ1969, both in spore and vegetative forms. The obtained results revealed that bacterial spores germinated in all volunteers. SNZ1969 persistence in the gut was monitored for two weeks after the end of gummy candy consumption, indicating its potential for prolonged colonization. These findings highlight the potential of unconventional food carriers for probiotic delivery and suggest that spore-forming probiotics can be metabolically active in the human intestine. These findings provide information for the development of food products containing spore-forming probiotics and their potential benefits in promoting gastrointestinal health.

Spore-forming probiotics Weizmannia coagulans isolated from human breast milk improves on the flavor development using micro-cheese platform

Weizmannia coagulans (formerly Bacillus coagulans) is a gram-positive bacterium forming spores and producing lactic acid, employing spore formation as a survival strategy within the host's intestinal and functioning as a probiotic. Therefore, we evaluated the probiotic functionalities of W. coagulans SRCM121381 and SRCM214335, isolated from human breast milk. Both strains demonstrated a remarkable tolerance capacity to acid and bile, and heat resistance. Moreover, we performed on Fermenter for Intestine Microbiome Model (FIMM) to evaluate the effects of these strains on the in vitro gut microbial fermentation based on a metagenomic approach. The FIMM analysis demonstrated that the addition of two W. coagulans strains significantly increased the population of beneficial bacteria, including Bifidobacterium and Blautia. Furthermore, to investigate the potential of two strains in the dairy food industry, we inoculated them into the production of Gouda cheese with a micro-cheese platform. These strains exhibited an enhancement of acid group flavors, accompanied by a reduction of undesirable flavors, such as bitterness, alcohol notes, and alkane group flavors. These results indicated that W. coagulans strains could accelerate cheese maturation and be used as an adjunct starter to facilitate flavor development in dairy products. Collectively, we proposed that the probiotic functionality of W. coagulans SRCM121381 and SRCM 214335 could influence intestinal microbial modulation by the increment of beneficial bacterial populations. And, demonstrated that these strains could applicate as adjunct starters in Gouda cheese production, leading to improved flavor profiles. These findings deepen our understanding of W. coagulans as potential probiotic bacteria in the dairy food industry.

Novel chitinolytic Bacillus spp. increase feed efficiency, feed digestibility, and survivability to Vibrio anguillarum in European seabass fed with diets containing Hermetia illucens larvae meal

Insect meals (IM) are being pointed out as a promising sustainable protein source. However, chitin in IM may negatively interfere with fish performance and nutrient availability. Given that carnivorous fish have a low or inexistent ability to digest chitin, novel strategies are needed to overcome chitin-imposed bottlenecks. Carbohydrolytic sporeforming probiotics offer a competitive industrial approach, being able to resist the harsh feed manufacturing process, transport, storage, and animal's gastrointestinal tract, while being able to increase the digestibility of otherwise indigestible components. Aiming to increase IM chitin digestibility, two chitinolytic sporeforming fish isolates (Fish isolates 645 (FI645) and 658 (FI658)), closely related to Bacillus licheniformis, were isolated from European sea bass (ESB) gastrointestinal tract, and incorporated (individually or as a mixture) in ESB diets with high defatted Hermetia illucens larvae meal (HM) levels (30% inclusion). FI were evaluated regarding their effects on ESB growth performance and digestibility. The spores were able to maintain their viability throughout the 180 days of diet storage at room temperature. Dietary inclusion of FI645 led to higher chitin digestibility. Concomitantly, the digestibility of dry matter, protein, and energy increased, overall leading to higher feed efficiency and protein efficiency ratio. Dietary inclusion of FI645 led to increased plasma N-acetylglucosamine (GlcNAc) levels. This is the first evidence for diet-mediated modulation of plasma GlcNAc concentrations. Dietary inclusion of FI645 also increased the expressions of the genes coding for N-acetylglucosamine kinase (nagk) and GlcNAc phosphomutase (pgm3), key enzymes from the GlcNAc salvage pathway and the hexosamine biosynthetic pathway (HBP). On the other hand, no differences were found for the expression of N-acetyl-D-glucosamine-6-phosphate deacetylase (amdhd2), a precursor enzyme for GlcNAc redirection to glycolysis. These results hint that free GlcNAc resulting from increased chitin digestibility might be preferentially redirected to protein O-GlcNAcylation processes through HBP in fish cells. Additionally, when challenged with Vibrio anguillarum, and after being feed for one month with the same diets, D645 also increased ESB survival from 52.5% to 77.5%, when compared to the control. Together, these results establish the potential of using FI645 as a probiotic for enhancing chitin digestion and acting as a prophylactic agent for ESB protection against V. anguillarum infection. This strategy opens novel possibilities for using commodities, including high chitin levels in aquafeeds, and reducing the use of antibiotics in aquaculture.

Effect of Spore-Forming Probiotics on the Poultry Production: A Review.

Due to the bad aspects associated with the use of antibiotics, the pressure on poultry production prompted the efforts to find out suitable growth-promoting and disease-preventing alternatives. Although many cost-effective alternatives have been developed, currently, one of the most auspicious alternatives for poultry feed is spore-forming probiotics, which can exert more beneficial effects as compared to normal probiotics, because of their ability to withstand the harsh external and internal conditions which result in increased viability. Many studies have already used spore-forming probiotics to improve different parameters of poultry production. Our laboratory has recently isolated a spore-forming bacterial strain, which has the potential to be used as a probiotic. So, to provide a detailed understanding, the current review aimed to collect valuable references to describe the mechanism of action of spore-forming probiotics and their effect on all the key aspects of poultry production.

The probiotic Bacillus subtilis BS50 decreases gastrointestinal symptoms in healthy adults: a randomized, double-blind, placebo-controlled trial

ABSTRACT Durable spore-forming probiotics are increasingly formulated into foods, beverages, and dietary supplements. To help meet this demand, the safety and efficacy of daily supplementation of Bacillus subtilis BS50 for 6 weeks was investigated in a randomized, double-blind, placebo-controlled, parallel clinical trial of 76 healthy adults. Before and during supplementation, gastrointestinal symptoms were recorded daily using a multi-symptom questionnaire. Clinical chemistry, hematology, plasma lipids, and intestinal permeability and inflammation markers were measured at baseline and end of study. Compared to placebo, 2 × 109 colony-forming units (CFU) BS50 per day increased the proportion of participants showing improvement from baseline to week 6 in the composite score for bloating, burping, and flatulence (47.4% vs. 22.2%), whereby the odds of detecting an improvement were higher with BS50 (OR [95% CI]: 3.2 [1.1, 8.7], p = .024). Analyses of individual gastrointestinal symptoms indicate that BS50 increased the proportion of participants showing an improvement at week 6 compared to placebo for burping (44.7% vs. 22.2%, p = .041) and bloating (31.6% vs. 13.9%, p = .071), without affecting other symptoms. There were no clinically meaningful changes in clinical chemistry, hematology, plasma lipids and intestinal permeability and other inflammation markers. In conclusion, the results suggest that dietary supplementation of 2 × 109 CFU Bacillus subtilis BS50 per day is a well-tolerated and safe strategy to alleviate gas-related gastrointestinal symptoms in healthy adults. Abbreviations AE adverse event; BHD bowel habits diary; BMI body mass index; BSS Bristol Stool Scale; CFU colony-forming unit; CRP C-reactive protein; FGID functional gastrointestinal disorder; GI gastrointestinal; GITQ Gastrointestinal Tolerance Questionnaire; GLP-1 glucagon-like peptide 1; GSRS Gastrointestinal Symptom Rating Scale; HDL-C high-density lipoprotein-cholesterol; IBS irritable bowel syndrome; IL-10 interleukin-10; ITT intent-to-treat; LBP lipopolysaccharide binding protein; LDL-C low-density lipoprotein-cholesterol; PP per protocol; PYY peptide YY; TG triglyceride; total-C total cholesterol

Современные спорообразующие пробиотики в лечении нарушений микробиоценоза кишечника: эффективность и безопасность

Приведены данные литературы, характеризующие механизмы действия, эффективность и безопасность спорообразующих бактерий, которые используются в качестве пробиотиков. Изложено влияние пробиотика-биоэнтеросептика энтерожермины, содержащей полиантибиотикорезистентные споры штамма Bacillus clausii, на нарушения микрофлоры кишечника, состояние иммунной системы. Положительными особенностями энтерожермины являются таксономическая идентификация, эффект самоэлиминации, антибиотико-резистентность, высокий профиль безопасности и возможность применения с целью профилактики побочных эффектов со стороны желудочно-кишечного тракта, обусловленных приѐмом антибиотиков и химиотерапевтических средств.

Efficacy and safety of spore-forming probiotics in the treatment of functional dyspepsia: a pilot randomised, double-blind, placebo-controlled trial

Current treatments for functional dyspepsia have limited efficacy or present safety issues. We aimed to assess spore-forming probiotics in functional dyspepsia as monotherapy or add-on therapy to long-term treatment with proton-pump inhibitors. In this single-centre, randomised, double-blind, placebo-controlled pilot trial that took place at University Hospitals Leuven (Leuven, Belgium), adult patients (≥18 years) with functional dyspepsia (as defined by Rome IV criteria, on proton-pump inhibitors or off proton-pump inhibitors) were randomly assigned (1:1) via computer-generated blocked lists, stratified by proton-pump inhibitor status, to receive 8 weeks of treatment with probiotics (Bacillus coagulans MY01 and Bacillus subtilis MY02, 2·5 × 109 colony-forming units per capsule) or placebo consumed twice per day, followed by an open-label extension phase of 8 weeks. Individuals with a history of abdominal surgery, diabetes, coeliac or inflammatory bowel disease, active psychiatric conditions, and use of immunosuppressant drugs, antibiotics, or probiotics in the past 3 months were excluded. All patients and on-site study personnel were masked to treatment allocation in the first 8 weeks. Symptoms, immune activation, and faecal microbiota were assessed and recorded. The primary endpoint was a decrease of at least 0·7 in the postprandial distress syndrome (PDS) score of the Leuven Postprandial Distress Scale in patients with a baseline PDS score of 1 or greater (at least mild symptoms), assessed in the intention-to-treat population. This study is registered with ClinicalTrials.gov, NCT04030780. Between June 3, 2019, and March 11, 2020, of 93 individuals assessed for eligibility, we included 68 patients with functional dyspepsia (51 [75%] women, mean age 40·1 years [SD 14·4], 34 [50%] on proton-pump inhibitors). We randomly assigned 32 participants to probiotics and 36 to placebo. The proportion of clinical responders was higher with probiotics (12 [48%] of 25) than placebo (six [20%] of 30; relative risk 1·95 [95% CI 1·07-4·11]; p=0·028). The number of patients with adverse events was similar with probiotics (five [16%] of 32) and placebo (12 [33%] of 36). Two serious adverse events occurring during the open-label phase (appendicitis and syncope in two separate patients) were assessed as unlikely to be related to the study product. In this exploratory study, B coagulans MY01 and B subtilis MY02 were efficacious and safe in the treatment of functional dyspepsia. Participants had potentially beneficial immune and microbial changes, which could provide insights into possible underlying mechanisms as future predictors or treatment targets. MY HEALTH.

Spore-forming probiotics for functional dyspepsia

Functional dyspepsia is a gastroduodenal disorder affecting nearly 10% of the global population,1 yet the condition shows no structural or biochemical abnormalities. Instead, functional dyspepsia is a disorder of motility and sensation with limited effective treatment options beyond first-line acid suppression with proton-pump inhibitors. The causes of functional dyspepsia are unknown, but evidence suggests a disruption of the homoeostatic balance between the gastrointestinal immune system, microbiota, and host physiology.

Molecular screening of bacteriocin-producing Bacillus spp. isolated from Soumbala, a fermented food condiment from Parkia biglobosa seeds

Bacteriocin-producing Bacillus species have an attractive interest as less studied spore-forming probiotics for food production. In this study, 20 strains of Bacillus were isolated from soumbala, a local fermented food condiment from Burkina Faso. They were characterized based on phenotype, antibacterial activity against Listeria monocytogenes NTCT983 and genotypic analysis by PCR, using specific primers for Bacillus sp., B. subtilis group, B. cereus group, bacteriocin genes, subtilin gene (spaS) and subtilosin gene (sboA). All the isolates were presumptively identified as Bacillus spp. with predominance of B. subtilis species (30%). Out of 20 strains, 11 showed anti-Listeria activity, with inhibition zones ranging from 11.12±0.53 mm (strain B21) to 25.25±0.35 mm (strain F25). PCR profiles revealed that 18 strains belonged to the B. subtilis group while O28 and O46 belonged to the B. cereus species. In addition, 13 strains carried bacteriocin genes, 10 possessed gene sboA and only strain O48 carried gene spaS. This finding highlights the predominance of bacteriocin-producing B. subtilis group species in Soumbala production population. These could be further characterized as probiotic starter cultures carrier for controlled production of high-quality soumbala.

A Review of the Effects and Production of Spore-Forming Probiotics for Poultry.

Simple SummarySpore-forming probiotics are widely used in the poultry industry for their beneficial impact on host health. The main feature that separates spore-forming probiotics from the more common lactic acid probiotics is their high resistance to external and internal factors, resulting in higher viability in the host and correspondingly, greater efficiency. Their most important effect is the ability to confront pathogens, which makes them a perfect substitute for antibiotics. In this review, we cover and discuss the interactions of spore-forming probiotic bacteria with poultry as the host, their health promotion effects and mechanisms of action, impact on poultry productivity parameters, and ways to manufacture the probiotic formulation. The key focus of this review is the lack of reproducibility in poultry research studies on the evaluation of probiotics’ effects, which should be solved by developing and publishing a set of standard protocols in the professional community for conducting probiotic trials in poultry.One of the main problems in the poultry industry is the search for a viable replacement for antibiotic growth promoters. This issue requires a “one health” approach because the uncontrolled use of antibiotics in poultry can lead to the development of antimicrobial resistance, which is a concern not only in animals, but for humans as well. One of the promising ways to overcome this challenge is found in probiotics due to their wide range of features and mechanisms of action for health promotion. Moreover, spore-forming probiotics are suitable for use in the poultry industry because of their unique ability, encapsulation, granting them protection from the harshest conditions and resulting in improved availability for hosts’ organisms. This review summarizes the information on gastrointestinal tract microbiota of poultry and their interaction with commensal and probiotic spore-forming bacteria. One of the most important topics of this review is the absence of uniformity in spore-forming probiotic trials in poultry. In our opinion, this problem can be solved by the creation of standards and checklists for these kinds of trials such as those used for pre-clinical and clinical trials in human medicine. Last but not least, this review covers problems and challenges related to spore-forming probiotic manufacturing.

Orange Juice and Yogurt Carrying Probiotic Bacillus coagulans GBI-30 6086: Impact of Intake on Wistar Male Rats Health Parameters and Gut Bacterial Diversity.

This study aimed to investigate the impact of the food matrix (orange juice and yogurt) on the effects of the spore-forming probiotic microorganism Bacillus coagulans GBI-30 6086 in health parameters and gastrointestinal tract (gut) bacterial diversity in Wistar male rats. Rats (n = 48) were randomly distributed into six groups. The groups were the Control (which received sterile distilled water), Juice (which received orange juice), Yogurt (which received yogurt), Probiotic Bacillus (which received B. coagulans GBI-30 6086 in distilled water), Probiotic Juice (which received orange juice with B. coagulans GBI-30 6086), and Probiotic Yogurt (which received yogurt with B. coagulans GBI-30 6086). Each animal belonging to the different groups was treated for 21 days. The daily administration of probiotic juice or probiotic yogurt did not affect the rats’ food or body weight. Rats fed with Probiotic Yogurt showed lower glucose and triglycerides levels (p < 0.05) in comparison to the control group (p < 0.05), while no changes in these parameters were observed in the rats fed with Probiotic Juice. Rats fed with Probiotic Yogurt showed a higher gut bacterial diversity than the control group (p < 0.05), and higher abundance (p < 0.05) of Vibrionales, Enterobacteriales, Burkholderiales, Erysipelotrichales, and Bifidobacteriales compared to all other groups. No changes were observed in the expression levels of antioxidant enzymes or heat shock protein 70 of rats fed with probiotic yogurt or probiotic juice. Results reveal that the consumption of yogurt containing B. coagulans GBI-30 6086 decreases triglycerides and glucose levels and positively impacts the gut bacterial ecology in healthy rats. These animal model findings indicate that the matrix also impacts the functionality of foods carrying spore-forming probiotics. Besides, this research indicates that yogurt is also a suitable food carrier of Bacillus coagulans GBI-30 6086.

Antimicrobial effect of spore-forming probiotics Bacillus laterosporus and Bacillus megaterium against Listeria monocytogenes.

There are several reports on the detection of Listeria monocytogenes in Iran, which has been identified as a major public health problem. In addition, due to drug resistance and strong biofilm formation of this pathogen, new approaches such as using probiotics are needed to combat this bacterium. The aim of this study was to investigate the effect of spore-forming probiotics on the expression of virulence genes hly, plc, inlA, and invasion of L. monocytogenes. Spore-forming probiotics were co-cultured with L. monocytogenes at different time points and following cell count, the expression level of virulence genes of L. monocytogenes was assessed by real-time PCR. To investigate the effect of the isolated strains on the invasiveness of L. monocytogenes at different time points, HT-29 cell line was used. Sporulated probiotics reduced the growth of L. monocytogenes in broth medium at different time periods. The number of L. monocytogenes co-cultured with Bacillus laterosporus decreased by 1, 3, 3, and 6 log at 2, 4, 8 and 24h after treatment, respectively. In addition, the expression of virulence genes was decreased at different time points and the expression of hly and plcA genes was more affected. The invasion rate of L. monocytogenes on HT-29 cells was decreased by 1 to 6 log at different time points. Both probiotic strains inhibited bacterial growth at different time points; however, more sensitive methods should be established for an accurate assessment.

A Spore‐Forming Probiotic Supplement Improves the Intestinal Immune Response and Protects the Intestinal Health During Recurrent Clostridioides difficile Colonization in Mice

Around 15%-30% of patients develop recurrent Clostridioides difficile infection (CDI) as conventional therapies disrupt protective gut microbiota. We tested if supplementation with a spore-forming probiotic would protect intestinal health in a mouse model of recurrent CD colonization. Methods: Female CF-1 mice were exposed to CD spores (4-log10 colony-forming units/10 μL) and then randomly assigned to receive either saline (CD-S) or probiotic (CD-PRO). Control mice received only saline (control). Following confirmation of initial CD colonization, mice were treated with vancomycin (10 days). After 5 days, mice recolonized with CD were treated again with vancomycin (10 days) and euthanized 5 days later. Fecal samples were collected at select time points for bacterial analysis. Following euthanasia, blood samples, cecum contents, and the intestine were collected for analysis. Probiotic supplementation mitigated the antibiotic-induced changes in cecum weight (P < .001). Probiotic-supplemented mice had increased messenger RNA expression of several immune parameters, accompanied by lower serum iron levels compared with CD-S mice (P < .05). Lower expressions of TNF α and calprotectin (P ≤ .05) were observed in CD-PRO mice compared with CD-S. The probiotics also supported the expression of intestinal tight junction proteins, which were diminished in the proximal colon of CD-S mice (P < .05). Mice supplemented with targeted spore-forming probiotics exhibited improved immune responses and nutrition immunity properties, which were linked with less inflammation and enhanced intestinal barrier proteins during recurrent CD colonization.

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.

Efficacy of Bacillus subtilis and bacitracin methylene disalicylate on growth performance, digestibility, blood metabolites, immunity, and intestinal microbiota after intramuscular inoculation with Escherichia coli in broilers

This experiment was conducted to evaluate the effect of Bacillus subtilis (BS) on broiler performance and health after intramuscular inoculation with E. coli and compare its effect with a growth promoter antibiotic. In a completely randomized design manner, 360 male Ross 308 chicks were divided into 6 treatments and 5 replicates of 12 chicks in each replicate. Experimental treatments included control diet, control + E. coli (0.5 mL of culture containing 108 CFU of E. coli/ml), control + 0.1% BS, control + 0.05% bacitracin methylene disalicylate (BMD), control + E. coli and BS, and control + E. coli and BMD in a factorial arrangement (3 × 2). Addition of BMD or BS to the control diet significantly (P < 0.01) increased body weight and decreased FCR, but E. coli challenge adversely reduced (P < 0.01) body weight and increased FCR, so that the addition of BMD or BS did not compensate growth reduction. E. coli challenged chicks had the lowest vaccine titers for ND, IB, AI, and IBD and the highest were observed in chicks fed BS. The E. coli challenge significantly (P < 0.01) increased albumin, globulin, cholesterol, triglyceride, LDL, ALT, and ALP indices. Addition of BMD and BS decreased albumin and globulin in challenged chick's plasma but had no effect on plasma lipid profile concentration. The E. coli challenge decreased villus height and increased crypt depth and goblet cell numbers significantly (P < 0.01). In birds subjected to BMD or BS, crypt depth decreased and villus height increased (P < 0.01), compared with the control diet. Challenge of E. coli significantly (P < 0.01) increased the bacterial population of E. coli, coliforms, and Salmonella in cecal parts of broilers’ intestines. In challenged birds receiving BMD or BS, E. coli, coliform, and Salmonella populations of ceca showed a significant (P < 0.01) reduction. Both BMD and BS increased the digestibility of nutrients significantly (P < 0.01), but a reduction was observed in E. coli challenged groups. Results of the study suggest that spore-forming probiotics are partially effective in unsuitable rearing situations such as colibacillosis in which the load of harmful bacteria is high.

Potential Spore-Forming Probiotics Isolated from Ntoba mbodi, Alkaline Fermented Leaves of Cassava from the Republic of the Congo

Potential Spore-Forming Probiotics Isolated from Ntoba mbodi, Alkaline Fermented Leaves of Cassava from the Republic of the Congo

Microbiological and molecular characterization of commercially available probiotics containing Bacillus clausii from India and Pakistan

Probiotics are actively used for treatment of diarrhoea, respiratory infections, and prevention of infectious gastrointestinal diseases. The efficacy of probiotics is due to strain-specific features and the number of viable cells; however, several reports of deviations from the label in the actual content of strains in probiotic products are a matter of concern. Most of the available data on quality focuses on probiotic products containing lactobacilli and/or bifidobacteria, while very few data are available on spore-forming probiotics. The present study evaluates the label claims for spore count and species identification in five commercial probiotic products marketed in India and Pakistan that claim to contain Bacillus clausii: Tufpro, Ecogro, Enterogermina, Entromax, and Ospor. Bacterial enumeration from three batches was done by microbiological plating methods by two independent operators. Species identification was done using PCR amplification and sequence analysis of the 16S rRNA gene, and determination of the total amount of species present in the products was done using PCR-denaturing gradient gel electrophoresis (PCR-DGGE) analysis followed by DNA sequencing of the excised bands. Plate count methods demonstrated poor correlations between quantitative label indications and bacteria recovered from plates for Tufpro, Ecogro, and Ospor. The 16S rRNA analysis performed on bacteria isolated from plate counts showed that only Enterogermina and Ospor contained homogenous B. clausii. PCR-DGGE analysis revealed that only Enterogermina had a homogenous B. clausii population while other products had mixed bacterial populations. In conclusion, the current analysis clearly demonstrates that of the five analysed commercial probiotics, only Enterogermina followed the label claims.

Tolerance to challenges miming gastrointestinal transit by spores and vegetative cells of Bacillus clausii

To study Bacillus clausii from a pharmaceutical product (Enterogermina O/C, N/R, SIN, T) and reference strains (B. clausii and Bacillus subtilis) for eco-physiological aspects regarding the gut environment. Spores and vegetative cells were challenged in vitro miming the injury of gastrointestinal transit: pH variations, exposure to conjugated and free bile salts, microaerophilic and anaerobic growth. No relevant differences were found studying the growth at pH 8 and 10, whereas at pH 7 the yields obtained for O/C and SIN were higher than those obtained for N/R and T strains. The spores were able to germinate and grow in the presence of conjugated bile salts (up to 1%, w/v) or free bile salts (0.2%) and also exhibited tolerance for the combined acid-bile challenge. As evidenced by lag-time, growth rate and cell yield the tolerance of Enterogermina isolates for conjugated salts was comparable with that of B. clausii type strain (DSM 8716(T)), and resulted higher than that observed for B. subtilis (ATCC 6051(T)). All the considered B. clausii strains demonstrated microaerophilic growth, but only some grew anaerobically in a nitrate medium. The ability of B. clausii spores to germinate after an acid challenge and grow as vegetative cells both in the presence of bile and under limited oxygen availability is consistent with the beneficial health effects evidenced for spore-forming probiotics in recent clinical studies. The experimental evidence from this study emphasizes some functional properties of B. clausii strains regarding their use as probiotics.