Promising clinical and immunological efficacy of Bacillus clausii spore probiotics for supportive treatment of persistent diarrhea in children.

Persistent diarrhea is a severe gastroenteric disease with relatively high risk of pediatric mortality in developing countries. We conducted a randomized, double-blind, controlled clinical trial to evaluate the efficacy of liquid-form Bacillus clausii spore probiotics (LiveSpo CLAUSY; 2 billion CFU/5 mL ampoule) at high dosages of 4-6 ampoules a day in supporting treatment of children with persistent diarrhea. Our findings showed that B. clausii spores significantly improved treatment outcomes, resulting in a 2-day shorter recovery period (p < 0.05) and a 1.5-1.6 folds greater efficacy in reducing diarrhea symptoms, such as high frequency of bowel movement of ≥ 3 stools a day, presence of fecal mucus, and diapered infant stool scale types 4-5B. LiveSpo CLAUSY supportive treatment achieved 3 days (p < 0.0001) faster recovery from diarrhea disease, with 1.6-fold improved treatment efficacy. At day 5 of treatment, a significant decrease in blood levels of pro-inflammatory cytokines TNF-α, IL-17, and IL-23 by 3.24% (p = 0.0409), 29.76% (p = 0.0001), and 10.87% (p = 0.0036), respectively, was observed in the Clausy group. Simultaneously, there was a significant 37.97% decrease (p = 0.0326) in the excreted IgA in stool at day 5 in the Clausy group. Overall, the clinical study demonstrates the efficacy of B. clausii spores (LiveSpo CLAUSY) as an effective symptomatic treatment and immunomodulatory agent for persistent diarrhea in children.Trial registration: NCT05812820.

Septicemia due to Bacillus clausii after the use of probiotics. A complication to keep in mind. / Septicemia por Bacillus clausii posterior al uso de probióticos. Una complicación para tener presente.

Probiotics are live microorganisms that benefit the host in different clinical situations. Bacillus clausii is one of the most frequently used, but it is not without risk. To date, there are few reports of complications secondary to this agent in pediatric patients. OBJECTIVE: To describe the case of an infant who developed after treatment sepsis due to Bacillus clausii. CLINICAL CASE: A 4-month-old female infant of indigenous ethnicity, from a rural area in the interior of Panama, 3 hours away from the nearest health sub-center by canoe, and with protein-calorie malnutrition, presented with acute diarrhea and moderate-severe dehydration, receiving Enterogermina as part of the initial treatment. She was transferred to a tertiary hospital, where she arrived with impaired consciousness, respiratory distress, and signs of shock. The initial blood culture reported growth of methicillin-resistant Staphylococcus aureus (MRSA), the gastrointestinal panel was positive for Clostridiodes difficile, and later serial blood cultures of peripheral blood and central venous catheter confirmed growth of Bacillus clausii. With a torpid evolution and resistance to multiple antibiotic schemes, she died due to multisystem organ failure twelve days after admission. CONCLUSIONS: The use of probiotics as concomitant treatment in patients with some degree of immunosuppression should be administered with caution, considering the presence of risk criteria for complications such as malnutrition or intestinal epithelial damage due to severe diarrhea since they predispose to the development of bacteremia and/or sepsis.

Structural insights into the DNA recognition mechanism by the bacterial transcription factor PdxR.

Specificity in protein-DNA recognition arises from the synergy of several factors that stem from the structural and chemical signatures encoded within the targeted DNA molecule. Here, we deciphered the nature of the interactions driving DNA recognition and binding by the bacterial transcription factor PdxR, a member of the MocR family responsible for the regulation of pyridoxal 5'-phosphate (PLP) biosynthesis. Single particle cryo-EM performed on the PLP-PdxR bound to its target DNA enabled the isolation of three conformers of the complex, which may be considered as snapshots of the binding process. Moreover, the resolution of an apo-PdxR crystallographic structure provided a detailed description of the transition of the effector domain to the holo-PdxR form triggered by the binding of the PLP effector molecule. Binding analyses of mutated DNA sequences using both wild type and PdxR variants revealed a central role of electrostatic interactions and of the intrinsic asymmetric bending of the DNA in allosterically guiding the holo-PdxR-DNA recognition process, from the first encounter through the fully bound state. Our results detail the structure and dynamics of the PdxR-DNA complex, clarifying the mechanism governing the DNA-binding mode of the holo-PdxR and the regulation features of the MocR family of transcription factors.

Alkalihalobacillus clausii (formerly Bacillus clausii) spores lessen antibiotic-induced intestinal injury and reshape gut microbiota composition in mice.

The antibiotic-induced intestinal injury (AIJ) is associated with diarrhoea and gastrointestinal discomfort. However, the pathological intestinal mechanisms and related side effects associated with antibiotic use/misuse may be counteracted by probiotics. This study aims to evaluate the effect and the protective mechanisms of a probiotic formulation containing Alkalihalobacillus clausii (formerly Bacillus clausii; BC) spores in an experimental model of AIJ. C57/Bl6J mice were orally challenged with a high dose of ceftriaxone for five days along with BC treatment which lasted up to the 15th day. Our results showed the beneficial effect of the probiotic in preserving colonic integrity and limiting tissue inflammation and immune cell infiltration in AIJ mice. BC increased tight junction expression and regulated the unbalanced production of colonic pro- and anti-inflammatory cytokines, converging toward the full resolution of the intestinal damage. These findings were supported by the histological evaluation of the intestinal mucosa, suggesting a potential restoration of mucus production. Notably, BC treatment increased gene transcription of the secretory products responsible for epithelium repair and mucus synthesis and normalized the expression of antimicrobial peptides involved in immune activation. Reconstruction of complex and diverse gut microbiota in antibiotic-induced dysbiosis was recorded upon BC supplementation. Specifically, the expansion of A. clausii, Prevotella rara and Eubacterium ruminatium drove intestinal microbiota rebalance by primarily impacting Bacteroidota members. Taken together, our data indicate that BC administration alleviates AIJ by multiple converging mechanisms leading to restoring gut integrity and homeostasis and reshaping microbiota composition.

Surfactin-like lipopeptides from Bacillus clausii efficiently bind to spike glycoprotein of SARS-CoV-2.

The coronavirus disease 2019 (COVID-19) rapidly spread across the globe, infecting millions and causing hundreds of deaths. It has been now around three years but still, it remained a serious threat worldwide, even after the availability of some vaccines. Bio-surfactants are known to have antiviral activities and might be a potential alternative for the treatment of SARS-CoV-2 infection. In the present study, we have isolated and purified, a surfactin-like lipopeptide produced by a probiotic bacterial strain Bacillus clausii TS. Upon purification and characterization with MALDI analysis, the molecular weight of the lipopeptide is confirmed as 1037 Da (similar to surfactin C) which is known to have antiviral activities against various enveloped viruses. Purified surfactin-like lipopeptide showed efficient binding and inhibition of SARS-CoV-2 spike (S1) protein, revealed by competitive ELISA assay. Further, we have explored the complete thermodynamics of the inhibitory binding of surfactin-like lipopeptide with S1 protein using isothermal titration calorimetric (ITC) assay. ITC results are in agreement with ELISA with a binding constant of 1.78 × 10-4 M-1. For further validation of the inhibitory binding of surfactin-like lipopeptide with S1 protein and its receptor binding domain (RBD), we performed molecular docking, dynamics, and simulation experiments. Our results suggested that surfactin could be a promising drug agent for the spike protein targeting drug development strategy against SARS-CoV-2 and other emerging variants.Communicated by Ramaswamy H. Sarma.

Evaluation of Bacillus clausii CSI08, Bacillus megaterium MIT411 and a Bacillus cocktail on gastrointestinal health: a randomised, double-blind, placebo-controlled pilot study.

In the present study, the safety, tolerance and impact of 1×109 cfu Bacillus clausii CSI08, 1×109 cfu Bacillus megaterium MIT411 and a probiotic cocktail containing Bacillus subtilis DE111®, Bacillus megaterium MIT411, Bacillus coagulans CGI314, and Bacillus clausii CSI08 with a total count of 2.0×109 cfu administered daily were assessed as compared with a maltodextrin containing placebo control. A total of 98 study participants received daily doses for 45 days, followed by a washout period of 2 weeks. A questionnaire to capture the incidence and duration of upper respiratory tract, urinary tract and/or gastrointestinal complaints and a diary to capture stool regularity and consistency was kept daily to record compliance throughout the 45 days. Faecal and blood samples were collected for microbiological and haematological analysis at the start and end of the treatment period. The probiotic cocktail significantly decreased the incidence of loose stools throughout the entire study. The recorded respiratory, urinary and gastrointestinal symptoms, defecation frequency and other stool consistency were not influenced. No clinically relevant changes in blood parameters, such as liver and kidney function and no serious adverse events appeared during and after administration. There were no changes in symptoms including sadness, irritability, energy, appetite, tension, stress, sleep, cardiovascular events, aches and pains, and dizziness as determined by a mood questionnaire administered to participants at baseline and at the end of the treatment period. Similarly, the measured inflammatory cytokines, antioxidant levels, cholesterol, triglycerides, free amino acids or minerals remained unaffected. There were no negative changes in alpha or beta diversity of the microbiota with any of the treatment groups. These promising data suggest that these treatments were safe and well tolerated, and further work with larger cohorts are justified to determine the efficacy of these potential probiotics in select demographic groups. Trial registration number with clinicaltrials.gov at NCT04758845.

Efficacy of Bacillus clausii in Pediatric Functional Constipation: A Pilot of a Randomized, Double-Blind, Placebo-Controlled Trial.

PURPOSE: To evaluate the efficacy of Bacillus clausii in the treatment of pediatric constipation. METHODS: A randomized, double-blind, placebo-controlled trial was conducted from January, 2021 to January, 2022 in children aged 1-5 years diagnosed with functional constipation according to Rome IV criteria. They were assigned to receive either B. clausii or placebo, once daily for four weeks. The primary out-come was treatment success (defined as ≥3 spontaneous stools per week and stool consistency grade ≥3 on Bristol stool chart). The secondary outcome was a comparison of stool frequency, consistency (defined by Bristol stool grade), and constipation-related symptoms. RESULTS: This trial enrolled 38 children (B. clausii, n=20 and placebo, n=18). At 4 weeks, no significant difference was noted in the treatment success between B. clausii and placebo groups [45% vs 56%; P=0.52). On within-group analyses, the mean (SD) of Bristol stool grade increased in both the B. clausii [1.7 (0.5) to 2.8 (1.2); P=0.003] and placebo [1.8 (0.5) to 2.8 (1.2); P=0.01] groups. Significant increases in the treatment success rate (22% to 56%, P=0.01) and mean stool frequency per week [3 (0.9) to 4.2 (1.7), P=0.01] were pronounced only in the placebo group. The frequency of painful defecation and large fecal mass were also significantly decreased in both the groups. No serious adverse events were observed. CONCLUSIONS: A 4-week course of B. clausii as the sole treatment was not more effective than a placebo for the management of functional constipation in children aged 1-5 years.

Possible Curative Effects of Boric Acid and Bacillus clausii Treatments on TNBS-Induced Ulcerative Colitis in Rats.

Crohn's disease (CD) and ulcerative colitis (UC) are two chronic relapsing inflammatory bowel diseases (IBD). Although there are several treatment options available to improve the symptoms of IBD patients, there is no effective treatment that provides a definitive solution. In the present study, we aim to investigate the antioxidative/anti-inflammatory effects of oral administration of boric acid and Bacillus clausii in a rat trinitrobenzenesulfonic acid (TNBS)-induced colitis model. The effects of boric acid and B. clausii were examined in serum and colon tissues with the help of some biochemical and histological analyses. Elevated inflammation and oxidative damage were found in the blood and colon tissue samples in the TNBS-induced group according to the complete blood count (CBC), tumor necrosis factor (TNF) alpha, interleukin-35 (IL-35), malondialdehyde (MDA), glutathione peroxidase (GPx), myeloperoxidase (MPO), nitric oxide (NO), and histological findings. Particularly, the highest IL-35 level (70.09 ± 12.62 ng/mL) in the combined treatment group, highest catalase activity (5322 ± 668.1 U/mg protein) in the TNBS-induced group, and lower relative expression of inducible nitric oxide synthase in the TNBS-induced group than the control group were striking findings. According to our results, it can be concluded that boric acid showed more curative effects, even if B. clausii probiotics was partially ameliorative.

Efficacy and Safety of the Adjuvant Use of Probiotic Bacillus clausii Strains in Pediatric Irritable Bowel Syndrome: A Randomized, Double-Blind, Placebo-Controlled Study.

OBJECTIVES: Current irritable bowel syndrome (IBS) treatments have limited efficacy and probiotics like Bacillus clausii (B. clausii) were found to be effective in the management of several gastrointestinal disorders. This phase III trial assessed the efficacy and safety of adding B. clausii (four strains: O/C, N/R, SIN, T), versus placebo, to conventional treatment of pediatric IBS in Mexico. METHODS: Patients aged 6-17 years 11 months with IBS (Rome IV) for at least 2 months were randomized to receive either B. clausii (oral suspension, total dose 4 billion spores/day) or placebo once daily for 8 weeks. All patients also received conventional treatment. The primary endpoint was the difference in the proportion of patients with clinical improvements at Week 8 (Global Assessment Questions [GAQ]). Secondary endpoints included responders by Subject's Global Assessment of Relief for Children with IBS (SGARC); number/consistency of stools; abdominal distention/bloating; abdominal pain/intensity; and IBS behavior. RESULTS: 73.6% (95% confidence interval [CI] 67.3-80.0; B. clausii n = 129) and 78.5% (95% CI 72.5-84.4; placebo n = 130) of patients had symptom improvement (p = 0.8182). For Week 8 SGARC, 19.2% (B. clausii) and 20.9% (placebo) reported complete symptom relief. Stool evaluations, bloating, abdominal pain/intensity, and IBS behavior were similar between groups. Both treatments were well tolerated. CONCLUSION: No significant differences in efficacy between B. clausii and placebo were demonstrated in addition to conventional treatment. The sample size calculation was based on an expected placebo/conventional treatment response of 30-40%. However, the actual treatment response observed was 80% and, thus, a study with larger population would be warranted. In addition, this study was conducted during the COVID-19 pandemic, when such controlled social conditions may have resulted in better diet, greater family stability, less psychological stress, and lower risk of infections exacerbating IBS, thereby improving symptoms in both groups. EUDRACT NUMBER: 2018-004519-31.

Quantitative and qualitative characterization of commercially available oral suspension of probiotic products containing Bacillus Clausii spores.

Probiotics contain beneficial live bacteria that confer several health benefits to the host. For the past 50 years, spore-forming Bacillus species have been used in the form of probiotics. Among these, Bacillus clausii strains are used for the management of acute and antibiotic-associated diarrhoea. In the present work, we have evaluated the asserted label information on randomly chosen commercial Bacillus clausii spore suspension of probiotic products. The quality and number of viable bacteria were evaluated based on the colony count, antibiotic resistance, and hemolytic activity assays. The colony fingerprinting and 16S rRNA gene-sequencing techniques were used to confirm the presence of a univariate strain (Bacillus clausii). Our results corroborated the label count of 2 × 109 CFU/5 mL in BACIPRO®, ENTEROGERMINA®, and TUFPRO® products. However, vegetative spore count was not found to match with the given label count in BENEGUT®, PROALANA-B®, ß-LOCK®, and PROCILLUS® Bacillus clausii brands. In the hemolytic activity assay, except for ß-LOCK®, the other 6 products showed gamma-hemolysis activity. Bacillus clausii isolated from all 7 probiotic products demonstrated resistance to several broad-spectrum antibiotics. The 16S rRNA gene-sequencing data detected genera of Bacillus and Bacillus clausii strain in the BACIPRO®, ENTEROGERMINA®, PROALANA-B®, BENEGUT®, and TUFPRO® products; however, Ralstonia mannitolilytica and Paenibacillus dendritiformis species were identified in ß-LOCK® and PROCILLUS®, respectively. As correct label information was observed only in BACIPRO®, ENTEROGERMINA®, and TUFPRO® products, it is proposed that a more stringent quality check would minimize the possibility of mismatch concerning the label information.

Randomized, direct comparison study of Saccharomyces boulardii CNCM I-745 versus multi-strained Bacillus clausii probiotics for the treatment of pediatric acute gastroenteritis.

BACKGROUND: The choice of an appropriate probiotic for pediatric acute gastroenteritis (PAGE) can be confusing. Our aim was to compare the efficacy and safety of 2 probiotics (Saccharomyces boulardii CNCM I-745 vs a 4-strain mixture of Bacillus clausii O/C, SIN, N/R, T) for the treatment of PAGE. METHODS: A 2-arm parallel, randomized trial recruited children (6 months to 5 years old) with mild-moderate acute diarrhea, from 8 centers in Argentina. A total of 317 children were enrolled and blindly randomized to 5 days of either S boulardii CNCM I-745 (n = 159) or a 4-strain mixture of B clausii (n = 158), then followed for 7 days post-probiotic treatment. A stool sample was collected at inclusion for pathogen identification. The primary outcome was duration of diarrhea defined as the time from enrollment to the last loose stool followed by the first 24-hour period with stool consistency improvement. Secondary outcomes included frequency of loose stools/day, severity of diarrhea, number reporting no diarrhea at Day 6, time-to-first formed stool, recurrence of diarrhea by study end (Day 12) and safety outcomes. RESULTS: Three hundred twelve (98%) children completed the study. S boulardii CNCM I-745 showed a significant reduction (P = .04) in the mean duration of diarrhea (64.6 hours, 95% confidence interval [CI] 56.5-72.8) compared to those given B clausii (78.0 hours, 95% CI 69.9-86.1). Both probiotics showed improvement in secondary outcomes and were well-tolerated. CONCLUSION: In this study, S boulardii CNCM I-745 demonstrated better efficacy than B clausii mix for reducing the duration of pediatric acute diarrhea.

In vitro and in vivo evaluation of Bacillus clausii against Schistosoma mansoni.

Experimental studies and clinical trials have been showing that probiotics are promising in the prevention and control of parasite infections. B. clausii, obtained from Enterogermina®, was cultured to obtain cell-free culture supernatant (CFS) and spores to evaluate its schistosomicidal effect in vitro and in vivo against Schistosoma mansoni, respectively. For in vitro and in vivo analysis mice were infected with 120 and 50 cercariae, respectively. Couples of adult worms, recovered on day 45 of infection, were exposed to CFS. The in vivo assay was performed for 100 days, where all animals were infected on the 30th day. Four experimental groups were formed, as follows: G1 - Saline solution from the 1st until the 100th day; G2 - B. clausii from the 1st until the 100th day; G3 - B. clausii from the 68th day (onset of oviposition) until the 100th day and G4 - PZQ (50 mg/Kg) from the 75th until the 79th day. In vitro, CFS of B. clausii does not caused mortality nor changed the motility on S. mansoni adult worms. G2 and G3 showed reduction of the 68.58 and 44.25% in the number of eggs eliminated in the feces and 34.29 and 53.6% and 22.8 and 48.49% the number of eggs trapped in the liver and intestine, respectively. Furthermore, in both therapeutic regimens G2 and G3, B. clausii increased the percentage of dead eggs in the intestinal tissue. B. clausii CFS, in vitro, does not showed action against S. mansoni and that treatment with B. clausii spores modulates favorably the parasitological parameters in the experimental infection of S. mansoni.

Bacillus clausii UBBC-07 in the symptom management of upper respiratory tract infections in children: a double blind, placebo-controlled randomised study.

In children, upper respiratory tract infections (URTIs) are one of the most common causes of infections which often require outpatient consultations with the doctor. The purpose of this study was to evaluate the effect of probiotic Bacillus clausii UBBC-07 on symptom management of URTIs in children. In this double blind, randomised, placebo-controlled study, 90 children (age 4-7 years) with URTIs were equally divided into two groups, the probiotic and placebo. The children were instructed to take B. clausii UBBC-07 spores (2×109 per 5 ml vial) or placebo suspension daily twice for three months. The total duration of the study was 6 months, 3 months treatment and 3 months follow-up period. The parameters assessed were the mean number of URTIs, duration and severity of URTIs, absenteeism from school/childcare and immunity parameters, such as immunoglobulin (Ig)M, IgG, IgE and salivary IgA levels. At the end of treatment, there was a significant decrease in the number, duration and severity of URTIs in the probiotic treated group as compared to the baseline and placebo. IgE levels were significantly decreased and salivary IgA levels were significantly increased in the probiotic treated group suggesting probiotic mediated Th1/Th2 immune homeostasis to alleviate URTIs in children. In conclusion, B. clausii UBBC-07 may help in the reduction of symptoms of URTIs. The trial was registered prospectively with the Clinical Trial Registry of India (CTRI Reg. No: CTRI/2018/08/015282).

Bacillus clausii for Gastrointestinal Disorders: A Narrative Literature Review.

The gut microbiota is intrinsically linked to human health; disturbances in microbial homeostasis are implicated in both intestinal and extraintestinal disorders. Probiotics are "live microorganisms that, when administered in adequate amounts, confer a health benefit on the host," and many commercial preparations comprising a diverse range of species are available. While probiotics have been much researched, better understanding of the probiotic effects and applications of species such as Bacillus clausii is warranted. In this narrative literature review, we review the characteristics and mechanisms of action supporting B. clausii as a probiotic and discuss the evidence from clinical studies evaluating B. clausii probiotics for the management of a variety of gastrointestinal disorders and symptoms in children and adults. Finally, we highlight the challenges of future research and the need for more robust and diverse clinical evidence to guide physicians in the clinical application of probiotics for gastrointestinal disorders and other conditions.

Gut immune homeostasis: the immunomodulatory role of Bacillus clausii, from basic to clinical evidence.

INTRODUCTION: The gut microbiota affects the development of the gut immune system in early life. Perturbations to microbiota structure and composition during this period can have long-term consequences on the health of the individual, through its effects on the immune system. Research in the last few decades has shown that probiotic administration can reverse these effects in strain- and environment-specific ways. Bacillus clausii (B. clausii) has been in use for many decades as a safe and efficacious probiotic, but its mode of action has not yet been completely elucidated. AREAS COVERED: In this review, we discuss how the gut immune system works, the factors that affect its functioning, and the plethora of research highlighting its role in various diseases. We also discuss the known modes of action of Bacillus probiotics, and highlight the preclinical and clinical evidence that reveal how B. clausii acts to bolster gut defense. EXPERT OPINION: We anticipate that the treatment and/or prevention of dysbiosis will be central to managing human health and disease in the future. Discovering the pathophysiology of autoimmune diseases, infections, allergies, and some cancers will aid our understanding of the key role played by microbial communities in these diseases.

Insights into the mechanism for the high-alkaline activity of a novel GH43 ß-xylosidase from Bacillus clausii with a promising application to produce xylose.

Nowadays, alkali-tolerant ß-xylosidases and their molecular mechanism of pH adaptability have been poorly studied. Here, a novel GH43 ß-xylosidase (XYLO) was isolated from Bacillus clausii TCCC 11004, and the recombinant ß-xylosidase (rXYLO) was most active at pH 8.0 and stable in a broad pH range (7.0-11.0), exhibiting superior alkali tolerance. Molecular dynamics simulation indicated that XYLO showed a notable overall structural stability and an enlargement of substrate binding pocket under alkaline condition, resulting in the formation of a new hydrogen bond between substrate and Arg286 of XYLO, and the tight binding played a key role in improving the XYLO activity with the increasing pH. Moreover, rXYLO with an endo-xylanase resulted in high xylose yields by hydrolyzing alkali-extracted xylan from agricultural wastes. This work would provide an alkali-tolerant ß-xylosidase, enhance the understanding for the relationship of structure and activity adapted to the high-alkaline environment, and promote its application in xylose production.

The Comparative Effects of Inulin and Bacillus clausii on LPS-Induced Endotoxemic Rat Liver.

BACKGROUND: This paper sought to investigate the modifies of inulin and Bacillus clausii on the lipopolysaccharides (LPS) inducing oxidative stress signaling pathway in the endotoxemic rat model. METHODS: Wistar albino male rats (n = 36), divided into six groups, were formed randomly in the following stages: the control group; the prebiotic group (Inulin; 500 mg/kg); the probiotic group (Bacillus clausii; 1x109 CFU); the LPS group (1.5 mg/kg) as the endotoxemic model; the prebiotic group + LPS; and the probiotic group + LPS as treatment groups. RESULTS: The reactive oxygen species (ROS), advanced oxidation products of protein (AOPP), thiobarbituric acid reactive substances (TBARS), total oxidant status (TOS), oxidative stress index (OSI), and myeloperoxidase activity (MPO) levels increased in LPS-induced toxicity. Prebiotic treatment decreased LPS-induced hepatotoxicity on rat liver as observed in the decrease in the levels of oxidative stress parameters, such as ROS, TBARS, TOS, and OSI. The effect of the probiotic treatment on the ROS, AOPP, TOS, OSI levels was not statistically significant. However, it was determined that probiotic application was effective in the TBARS, TAS, and GSH levels. When the biochemical results of the prebiotic and probiotic treatment applications were compared, it was found that the prebiotic treatment was more effective on oxidative stress parameters (ROS, TBARS, TOS, and OSI). In addition, the histological damage score and MPO-staining results of the prebiotic treatment group were found to be more effective than the probiotic group. CONCLUSION: In this first study, where inulin and Bacillus clausii spores are used against liver damage caused by LPS, inulin provides much more effective protection than Bacillus clausii spores.

Novel Detection Method for Evaluating the Activity of an Alkaline Serine Protease from Bacillus clausii.

Until now, the detection methods for serine proteases have been quite time-consuming or cannot indicate the "real" protease activity. Here, a rapid and simple method for determining the "real" activity of serine proteases toward AAPX (a kind of mixed polypeptide substrates, with X representing 20 standard amino acids) was developed. This AAPX method has high reliability, sensitivity, and repeatability and can be used for detecting the serine protease activity spectrophotometrically. Additionally, the site-directed saturation mutagenesis library of alkaline serine protease PRO (BcPRO) from Bacillus clausii was screened with this AAPX method. Three beneficial mutants S99R, S99H, and S99W were identified, and S99W displayed the highest activity. In comparison to wild-type BcPRO, S99W exhibited enhanced catalytic performance toward eight AAPX monomers, and the molecular dynamics simulation revealed the mechanism responsible for its improved activity toward AAPM. Consequently, this work provides an efficient method for detecting, characterizing, mining, and high-throughput screening of serine proteases.

Efficacy of Bacillus clausii UBBC - 07 spores in the amelioration of oral mucositis in head and neck cancer patients undergoing radiation therapy.

BACKGROUND: The natural flora of healthy mucosa offer protection to the host. The loss of this barrier during radiotherapy enhances insults from physical, chemical and microbial agents. METHODOLOGY: A randomized, double blind, placebo-controlled, parallel study on forty-six patients who underwent radiotherapy for head and neck cancers was undertaken. Patients were randomized either to standard treatment plus Bacillus clausii UBBC07 or standard treatment plus placebo. Bacillus clausii UBBC07 was given as an oral suspension of 2 billion spores twice every day for 30 days or until completion of total fractions of radiation. Grading of the mucositis was performed using CTCAE v.4.03 severity scale. The time taken for the appearance, resolution and severity of mucositis was evaluated. RESULTS: There was a significant increase (p < 0.01) in median time for the onset of mucositis i.e., 10 days in test and 8 days in control groups respectively. The median time for remission was found to be 12 days in test and 14 days in the control group (p < 0.05). Grade IV mucositis was observed in no patients in test group and 2 patients in the control group (p < 0.05). No adverse events attributed to the Bacillus clausii were seen. Bacillus clausii UBBC07 therapy delayed the onset, decreased the time to remission and displayed strong impact on suppressing the occurrence of high-grade mucositis amongst the test group. CONCLUSIONS: This study provides a positive trend that probiotics like Bacillus clausii UBBC07 spores could act as a tool to ameliorate oral mucositis.