DOCSLIB.ORG

Clostridium Difficile Infection; Establishment, Prevention and the Role of the Microbiota

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Clostridium Difficile Infection; Establishment, Prevention and the Role of the Microbiota Clostridium difficile infection; establishment, prevention and the role of the microbiota William Thomas Ferreira Thesis submitted to Royal Holloway, University of London in fulfilment of the requirements for the degree of Doctor of Philosophy School of Biological Sciences Royal Holloway, University of London 09 September 2019 1 Declaration of Authorship I declare that this work was carried out in accordance with the Regulations of the University of London. I declare that this submission is my own work, and to the best of my knowledge does not represent the works of others, published or unpublished, except where duly acknowledged in the text. No part of this thesis has been submitted for a higher degree at another university or institution. Signed: Date: 09/09/2019 William Thomas Ferreira Royal Holloway, University of London 2 Acknowledgements Firstly, I would like to thank my supervisor Prof. Simon Cutting who has provided me with continuous support, encouragement and inspiration. Also thank you to Hong, who has been a friend to me throughout my PhD and has made the joint projects we have worked on together so enjoyable. Many thanks also to Mikhail, you’ve provided me with many tools so useful in science, and I feel that my horizons have expanded by working with you. A big thank you to every member of the Cutting lab over the last 4 to 5 years, I would love to name all of you, but the list would be longer than my thesis… Of course, I couldn’t have done any of this without the loving support from my family, especially my Dad and my two amazing sisters. Thanks to all of my friends away from the lab for the never-ending support, you know who you are! Last but foremost I would like to thank my wife Suzie for her constant love and support, for the early mornings and late nights and for being the only thing keeping me sane the past few months. Thank you for being my editor, my audience, my collaborator and my muse. But most importantly, thanks for being my best friend. 3 Abstract Clostridium difficile is a leading cause of nosocomial, antibiotic-associated, diarrhoea in industrialised countries. C. difficile spores are dormant and resistant structures responsible for the colonisation and persistence of C. difficile infection (CDI) within patients as well as the transmission between them. This thesis examines firstly, the role of a C. difficile spore coat protein, CotE, in the establishment of CDI, and secondly, the role of allochthonous Bacillus within the GI tract in suppressing CDI. CotE is a protein displayed on the C. difficile spore surface which carries two functional elements, an N-terminal peroxiredoxin and a C-terminal chitinase domain. Using isogenic mutants, it was demonstrated in vitro and ex vivo that CotE enables binding of spores to mucus by direct interaction with mucin and contributes to its degradation. In animal models of CDI, it was shown that when CotE was absent, both colonisation and virulence were markedly reduced. It is demonstrated here that the attachment of spores to the intestine is essential in the development of CDI. The developments of infection within the host GI tract requires germination of the spore, followed by outgrowth of the vegetative cell; a process which requires a multitude of factors to favour C. difficile proliferation. In a series of experiments, it was shown that environmentally acquired Bacillus help to suppress CDI by the production of lipopeptides in vivo with activity against C. difficile and that the use of antibiotics abrogates this phenomenon. Using hamster and mouse models, it was demonstrated that the administration of these Bacillus provides complete colonisation resistance against CDI in clindamycin treated animals. Reversed-phase high-performance liquid chromatography, dynamic light scattering and MALDI-TOF analysis identified the active molecules to be a micellar complex of lipopeptides comprised of chlorotetaine, iturins, fengycins and surfactins. Through this work, it is demonstrated that the attachment of spores to the intestine is essential in the development of CDI and suggests a direct role for the spore in the establishment and promotion of disease. 4 Table of Contents Declaration of Authorship .................................................................................... 2 Acknowledgements ................................................................................................ 3 Abstract .................................................................................................................. 4 Table of Contents .................................................................................................. 5 List of Figures ...................................................................................................... 10 List of Tables ....................................................................................................... 13 Abbreviations ...................................................................................................... 14 1. Introduction ..................................................................................................... 17 1.1 Identification of C. difficile as a pathogen .................................................. 17 1.2 C. difficile infection ..................................................................................... 20 1.2.1 The disease ........................................................................................... 20 1.2.2 Presentation of the disease ................................................................... 23 1.2.3 Establishment of infection ................................................................... 24 1.2.4 Virulence factors .................................................................................. 28 1.2.4.1.Toxins……………………………….....……………………...….28 1.2.4.2 S-layer proteins…………………………………………………..33 1.2.5 Rise of community acquired infections ............................................... 34 1.3 The spore ..................................................................................................... 35 1.3.1 Spore structure and function ................................................................ 35 1.3.2 Sporulation pathway............................................................................. 40 1.3.3 Germination pathway ........................................................................... 42 1.4 Control and prevention of CDI ................................................................... 45 1.4.1 Diagnosis .............................................................................................. 45 1.4.2 Prevention ............................................................................................ 46 1.4.2.1 Hospital management and hygiene……………………………....46 5 1.4.2.2 Probiotics………………………………………………………...47 1.4.2.3 Developing a vaccine…………………………………………….49 1.4.3 Treatment ............................................................................................. 51 1.4.3.1.Antibiotics………………………………………………………..51 1.4.3.2 Faecal microbiota transplantation (FMT)………………………..51 1.5 The gut microbiome .................................................................................... 53 1.5.1 Role of the microbiota .......................................................................... 53 1.5.2 Effect of antibiotics .............................................................................. 58 1.5.3 Control of CDI ..................................................................................... 59 2. Materials and Methods ................................................................................... 62 2.1 General methods.......................................................................................... 62 2.2 Bacterial strains ........................................................................................... 62 2.3 Sporulation of C. difficile ............................................................................ 63 2.4 Spore purification ........................................................................................ 65 2.5 Spore coat extractions ................................................................................. 65 2.6 Antibody production ................................................................................... 65 2.7 Western blotting of spore coat proteins ...................................................... 65 2.8 Southern blotting ......................................................................................... 66 2.9 Germination assays ..................................................................................... 66 2.10 Spore adhesion to hydrocarbon (SATH) assay ......................................... 67 2.11 Cell culture ................................................................................................ 68 2.12 Cytotoxicity assays ................................................................................... 68 2.13 Induction of cytokines ............................................................................... 69 2.14 Spore adhesion assays ............................................................................... 70 2.15 Spore binding to pig mucus explants ........................................................ 70 2.16 Spore binding to ligands............................................................................ 71 6 2.17 Mucin degradation using amido black staining ........................................ 71 2.18 Mucin degradation using amido black staining
Load more

Recommended publications
  • APP204132 EPA Staff Advice Report.Pdf
    Staff Advice Report 13 October 2020 Application code: APP204132 Application type and sub-type: Statutory determination Applicant: Functional and Integrative Medicine Limited Date application received: 22 September 2020 Purpose of the Application: Information to support the consideration of the determination of Bacillus clausii and Bacillus indicus Purpose of this document 1. This document has been prepared by the EPA staff to advise the Committee of our assessment of application APP204132 submitted under the Hazardous Substances and New Organisms Act 1996 (the Act). This document discusses information provided in the application and other sources. 2. The decision path for this application can be found in Appendix 1. The application 3. With the fast expansion of the probiotic market, which is expected to reach $57.4 billion by 2022, the interest around the antimicrobial and immunomodulatory activities of Bacillus strains is quickly growing (Oyeniran 2019). The pharmaceutical industry is looking to develop probiotic supplements for animal feeds, as well as dietary supplements and registered medicines for humans (Hong et al. 2005; Cutting 2011; Patel 2011). 4. According to the applicant, probiotic products containing B. clausii and B. indicus have been imported into New Zealand for more than five years by consumers and practitioners. Functional and Integrative Medicine Limited (Fxmed) began to distribute a product containing the two bacteria two years ago. They only became aware of the restrictions around the import of new organisms in September 2020 when one of their shipments was held at the border by MPI. 5. On 22 September 2020, Fxmed applied to the EPA under section 26 of the HSNO Act seeking a determination on the new organism status of Bacillus clausii and B.
    [Show full text]
  • Bacillus Coagulans S-Lac and Bacillus Subtilis TO-A JPC, Two Phylogenetically Distinct Probiotics
    RESEARCH ARTICLE Complete Genomes of Bacillus coagulans S-lac and Bacillus subtilis TO-A JPC, Two Phylogenetically Distinct Probiotics Indu Khatri☯, Shailza Sharma☯, T. N. C. Ramya*, Srikrishna Subramanian* CSIR-Institute of Microbial Technology, Sector 39A, Chandigarh, India ☯ These authors contributed equally to this work. * [email protected] (TNCR); [email protected] (SS) a11111 Abstract Several spore-forming strains of Bacillus are marketed as probiotics due to their ability to survive harsh gastrointestinal conditions and confer health benefits to the host. We report OPEN ACCESS the complete genomes of two commercially available probiotics, Bacillus coagulans S-lac Citation: Khatri I, Sharma S, Ramya TNC, and Bacillus subtilis TO-A JPC, and compare them with the genomes of other Bacillus and Subramanian S (2016) Complete Genomes of Lactobacillus. The taxonomic position of both organisms was established with a maximum- Bacillus coagulans S-lac and Bacillus subtilis TO-A likelihood tree based on twenty six housekeeping proteins. Analysis of all probiotic strains JPC, Two Phylogenetically Distinct Probiotics. PLoS of Bacillus and Lactobacillus reveal that the essential sporulation proteins are conserved in ONE 11(6): e0156745. doi:10.1371/journal. pone.0156745 all Bacillus probiotic strains while they are absent in Lactobacillus spp. We identified various antibiotic resistance, stress-related, and adhesion-related domains in these organisms, Editor: Niyaz Ahmed, University of Hyderabad, INDIA which likely provide support in exerting probiotic action by enabling adhesion to host epithe- lial cells and survival during antibiotic treatment and harsh conditions. Received: March 15, 2016 Accepted: May 18, 2016 Published: June 3, 2016 Copyright: © 2016 Khatri et al.
    [Show full text]
  • Gut Microbiota Hypolipidemic Modulating Role in Diabetic Rats Fed with Fermented Parkia Biglobosa (Fabaceae) Seeds
    Gut Microbiota Hypolipidemic Modulating Role with Parkia biglobosa (Fabaceae) Seeds Vol. 11 (2), December 2020 Open Access ORIGINAL ARTICLE Full Length Article Gut Microbiota Hypolipidemic Modulating Role in Diabetic Rats Fed with Fermented Parkia biglobosa (Fabaceae) Seeds Olayinka Anthony Awoyinka1,*, Tola Racheal Omodara2, Funmilola Comfort Oladele1, Margret Olutayo Alese3, Elijah Olalekan Odesanmi4, David Daisi Ajayi5, Gbenga Sunday Adeleye6, Precious Bisola Sedowo2 1Department of Medical Biochemistry, College of Medicine, Ekiti State University Ado Ekiti Nigeria. 2Department of Microbiology, Faculty of Science, Ekiti State University, Ado Ekiti, Nigeria. 3Department of Anatomy, College of Medicine, Ekiti State University, Nigeria. 4Department of Biochemistry, Faculty of Science, Ekiti State University, Ado Ekiti, Nigeria. 5Department of Chemical Pathology, Ekiti State University Teaching Hospital, Ado Ekiti, Nigeria. 6Department of Physiology, College of Medicine, Ekiti State University, Ado Ekiti, Nigeria. ABSTRACT Background: Modulation and balancing of host gut microbiota by probiotics has been documented by several literature. Prebiotic diets such as locust beans have been known to encourage the occurrence of these beneficial microorganisms in the host gut. Objectives: To study the modulating role of gut microbiota in the hypolipidemic effect of fermented locust beans on diabetic Albino Wister rats as animal models. Methodology: Albino rats (Wistar strain), averagely weighing 125g were successfully induced with alloxan. There after this induction, anti-diabetic treatment was carried out on various groups of rats by feeding them ad libitum with a diet of milled fermented and unfermented Parkia biglobosa seeds, respectively. Results: After three weeks of treatment, it was observed that fermented locust beans caused a significant reduction (p ≤ 0.05) in glucose, total triglycerides, total cholesterol and LDL, while the HDL levels were significantly elevated (p ≤ 0.05).
    [Show full text]
  • Isolation and Characterization of Probiotic Bacillus Subtilis MKHJ 1-1 Possessing L-Asparaginase Activity
    applied sciences Article Isolation and Characterization of Probiotic Bacillus subtilis MKHJ 1-1 Possessing L-Asparaginase Activity Hyeji Lim 1 , Sujin Oh 1 , Sungryul Yu 2 and Misook Kim 1,* 1 Department of Food Science and Nutrition, Dankook University, Cheonan 31116, Korea; [email protected] (H.L.); [email protected] (S.O.) 2 Department of Clinical Laboratory Science, Semyung University, Jecheon 27136, Korea; [email protected] * Correspondence: [email protected]; Tel.: +82-41-550-3494 Abstract: The purpose of this study was to isolate functional Bacillus strains from Korean fermented soybeans and to evaluate their potential as probiotics. The L-asparaginase activity of MKHJ 1-1 was the highest among 162 Bacillus strains. This strain showed nonhemolysis and did not produce β-glucuronidase. Among the nine target bacteria, MKHJ 1-1 inhibited the growth of Escherichia coli, Pseudomonas aeruginosa, Shigella sonnei, Shigella flexneri, Klebsiella pneumoniae, Staphylococcus aureus, and Bacillus cereus. 16S rRNA gene sequence analysis resulted in MKHJ 1-1 identified as Bacillus subtilis subsp. stercoris D7XPN1. As a result of measuring the survival rate in 0.1% pepsin solution (pH 2.5) and 0.3% bile salt solution for 3 h, MKHJ 1-1 exhibited high acid resistance and was able to grow in the presence of bile salt. MKHJ 1-1 showed outstanding autoaggregation ability after 24 h. In addition, its coaggregation with pathogens was strong. Therefore, MKHJ 1-1 is a potential probiotic with L-asparaginase activity and without L-glutaminase activity, suggesting that it could be a new resource for use in the food and pharmaceutical industry.
    [Show full text]
  • Bacillus Clausii
    & P ga hys o ic Jayanthi and Ratna Sudha, J Yoga Phys Ther 2015, 5:4 Y a f l o T l h a e DOI: 10.4172/2157-7595.1000211 n r a r p u y o J Journal of Yoga & Physical Therapy ISSN: 2157-7595 ShortResearch Communication Article OpenOpen Access Access Bacillus clausii - The Probiotic of Choice in the Treatment of Diarrhoea Jayanthi N and Ratna Sudha M* Centre for Research and Development, Unique Biotech Ltd, R.R District, Hyderabad- 500 078, India Abstract Probiotics are known to have a role in enhancing digestive health. In this paper, evidence for the efficacy of Bacillus clausii, a spore forming probiotic (viz clinical studies) in the treatment of diarrhoea, prevention of antibiotic associated diarrhoea and in the prevention of side effects associated with Helicobacter pylori is presented. Mechanism of action suggested is through inhibition of pathogens and immunomodulatory effects. Bacillus clausii is the probiotic of choice in the treatment of diarrhoea as it has the added advantage of being a spore forming probiotic. It is therefore stable at room temperature and resistant to low pH ensuring that it reaches the small intestines where it can colonize and exert its beneficial effects. Keywords: Bacillus clausii; Diarrhea; Probiotic of the host against gastrointestinal tract diseases [15,16]. During acute diarrhoea, the normal gastrointestinal microbiota is found to Introduction undergo radical changes that facilitate the overgrowth of unwanted Probiotics are live microbes, which when administered in adequate microorganisms, including pathogenic strains. Several authors have amounts confer a health benefit to the host [1].
    [Show full text]
  • Mobile Antimicrobial Resistance Genes in Probiotics
    bioRxiv preprint doi: https://doi.org/10.1101/2021.05.04.442546; this version posted June 13, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Mobile antimicrobial resistance genes in probiotics Adrienn Greta´ Toth´ 1, Istvan´ Csabai2, Maura Fiona Judge3, Gergely Maroti´ 4,5, Agnes´ Becsei2,Sandor´ Spisak´ 5, and Norbert Solymosi3* 1Semmelweis University, Health Services Management Training Centre, 1125 Budapest, Hungary 2Eotv¨ os¨ Lorand´ University, Department of Phyisics of Complex Systems, 1117 Budapest, Hungary 3University of Veterinary Medicine Budapest, Centre for Bioinformatics, 1078 Budapest, Hungary 4Institute of Plant Biology, Biological Research Center, 6726 Szeged, Hungary 5University of Public Service, Faculty of Water Sciences, 6500 Baja, Hungary 6Department of Medical Oncology, Dana-Farber Cancer Institute, 02115 Boston, MA, USA *[email protected] ABSTRACT Even though people around the world tend to consume probiotic products for their beneficial health effects on a daily basis, recently, concerns were outlined regarding the uptake and potential intestinal colonisation of the bacteria that they transfer. These bacteria are capable of executing horizontal gene transfer (HGT) which facilitates the movement of various genes, including antimicrobial resistance genes (ARGs), among the donor and recipient bacterial populations. Within our study, 47 shotgun sequencing datasets deriving from various probiotic samples (isolated strains and metagenomes) were bioinformatically analysed. We detected more than 70 ARGs, out of which rpoB mutants conferring resistance to rifampicin, tet(W/N/W) and potentially extended-spectrum beta-lactamase (ESBL) coding TEM-116 were the most common.
    [Show full text]
  • Natural Strategies for Controlling Virulence and Antibiotic Resistance
    University of Connecticut OpenCommons@UConn Doctoral Dissertations University of Connecticut Graduate School 1-14-2019 Natural Strategies for Controlling Virulence and Antibiotic Resistance in Clostridium difficile Abraham Joseph Pellissery University of Connecticut - Storrs, [email protected] Follow this and additional works at: https://opencommons.uconn.edu/dissertations Recommended Citation Pellissery, Abraham Joseph, "Natural Strategies for Controlling Virulence and Antibiotic Resistance in Clostridium difficile" (2019). Doctoral Dissertations. 2036. https://opencommons.uconn.edu/dissertations/2036 Natural Strategies for Controlling Virulence and Antibiotic Resistance in Clostridium difficile Abraham Joseph Pellissery, PhD University of Connecticut, 2019 Clostridium difficile is a significant enteric pathogen causing a toxin-mediated infection and diarrhea in humans. There has been an increased incidence of C. difficile infection (CDI) in the United States with the emergence of hypervirulent strains and community associated outbreaks. CDI is commonly observed among hospital in-patients undergoing protracted antibiotic therapy, which results in gastrointestinal dysbiosis, creating a conducive environment for spore germination, pathogen colonization in the gut, and subsequent toxin production. An ideal anti- C.difficile therapeutic agent should inhibit critical virulence factors of the pathogen such as toxin production, sporulation and spore germination without inducing gut dysbiosis. Such agents when provided as an adjunct to C. difficile antibiotic therapy could help to improve the clinical outcome of CDI and prevent the relapse of the infection. In this Ph.D. research, the efficacy of three alternative treatment approaches as antivirulence agents was tested for potential future development as therapies against CDI. This included sodium selenite (metalloid), baicalin (flavone glycoside), and selected lactic acid bacteria.
    [Show full text]
  • The Putative Drug Efflux Systems of the Bacillus Cereus Group
    RESEARCH ARTICLE The putative drug efflux systems of the Bacillus cereus group Karl A. Hassan1,2☯, Annette Fagerlund3☯¤a, Liam D. H. Elbourne1, Aniko VoÈ roÈ s3, Jasmin K. Kroeger3,4, Roger Simm3¤b, Nicolas J. Tourasse3¤c, Sarah Finke3,5, Peter J. F. Henderson2, Ole Andreas Økstad3,5, Ian T. Paulsen1*, Anne-Brit Kolstø3,5* 1 Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney, NSW, Australia, 2 School of BioMedical Sciences and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, United Kingdom, 3 Laboratory for Microbial Dynamics (LaMDa), Section for Pharmaceutical Biosciences, School of Pharmacy, University of Oslo, Oslo, Norway, 4 Institut fuÈr Pharmazeutische Biologie a1111111111 und Biotechnologie, Albert-Ludwigs UniversitaÈt, Freiburg, Germany, 5 Centre for Integrative Microbial a1111111111 Evolution (CIME), Faculty of Mathematics and Natural Sciences, University of Oslo, 0316 Oslo, Norway a1111111111 a1111111111 ☯ These authors contributed equally to this work. a1111111111 ¤a Current address: Nofima, PB210, 1431 Ås, Norway ¤b Current address: Department of Oral Biology, Faculty of Dentistry, University of Oslo, OsloNorway ¤c Current address: ARNA Laboratory, INSERM U1212, CNRS UMR5320, Universite Bordeaux 2; 146 rue LeÂo-Saignat, Bordeaux, France * [email protected] (ABK); [email protected] (ITP) OPEN ACCESS Citation: Hassan KA, Fagerlund A, Elbourne LDH, Abstract VoÈroÈs A, Kroeger JK, Simm R, et al. (2017) The putative drug efflux systems of the Bacillus cereus The Bacillus cereus group of bacteria includes seven closely related species, three of group. PLoS ONE 12(5): e0176188. https://doi.org/ which, B. anthracis, B. cereus and B. thuringiensis, are pathogens of humans, animals and/ 10.1371/journal.pone.0176188 or insects.
    [Show full text]
  • Insecticidal Activity of Bacteria from Larvae Breeding Site with Natural Larvae Mortality: Screening of Separated Supernatant and Pellet Fractions
    pathogens Article Insecticidal Activity of Bacteria from Larvae Breeding Site with Natural Larvae Mortality: Screening of Separated Supernatant and Pellet Fractions Handi Dahmana 1,2 , Didier Raoult 1,2, Florence Fenollar 2,3 and Oleg Mediannikov 1,2,* 1 IRD, AP-HM, MEPHI, Aix Marseille University, 13005 Marseille, France; [email protected] (H.D.); [email protected] (D.R.) 2 IHU-Méditerranée Infection, 13005 Marseille, France; fl[email protected] 3 IRD, AP-HM, SSA, VITROME, Aix Marseille University, 13005 Marseille, France * Correspondence: [email protected]; Tel.: +33-(0)4-13-73-24-01; Fax: +33-(0)4-13-73-24-02 Received: 15 May 2020; Accepted: 17 June 2020; Published: 18 June 2020 Abstract: Mosquitoes can transmit to humans devastating and deadly pathogens. As many chemical insecticides are banned due to environmental side effects or are of reduced efficacy due to resistance, biological control, including the use of bacterial strains with insecticidal activity, is of increasing interest and importance. The urgent actual need relies on the discovery of new compounds, preferably of a biological nature. Here, we explored the phenomenon of natural larvae mortality in larval breeding sites to identify potential novel compounds that may be used in biological control. From there, we isolated 14 bacterial strains of the phylum Firmicutes, most of the order Bacillales. Cultures were carried out under controlled conditions and were separated on supernatant and pellet fractions. The two fractions and a 1:1 mixture of the two fractions were tested on L3 and early L4 Aedes albopictus. Two concentrations were tested (2 and 6 mg/L).
    [Show full text]
  • Safety of Micro-Organisms
    EUROPEAN COMMISSION HEALTH & CONSUMER PROTECTION DIRECTORATE-GENERAL Directorate C - Scientific Opinions C2 - Management of scientific committees II; scientific co-operation and networks Opinion on the use of certain micro-organisms as additives in feedingstuffs Expressed, 26 September 1997 Updated 25 April 2003 TERMS OF REFERENCE (September 1996) The Scientific Committee for Animal Nutrition is requested to give an opinion on the following questions: 1. Is the use of the micro-organisms shown in the annexed list1 safe to corresponding animal species under the conditions proposed? 2. Can their use result in development of resistance in bacteria to prophylactic or therapeutic preparations or exert an effect on the persistence of bacteria in the digestive tract of corresponding animal? Is or can the micro-organism become resistant to antibiotics? 3. Do the products indicated in the annexed list contain or consist of genetically modified organisms within the meaning of Article 2-1 and 2-2 of Council Directive 90/220/EEC2? If it is the case, was a specific environmental risk assessment carried out, similar to that laid down in the above-mentioned Directive, is the outcome satisfactory in view of the requirements of this Directive? 4. Do the toxicology studies allow to conclude that the proposed use does not present risks to the consumers, to the users ? 5. In the light of the answer to the above-mentioned questions, are the proposed con- ditions of use acceptable? 1 The List – not attached to this opinion - contained a list of those micro organisms submitted at the time of the original question. 2 On the deliberate release into the environment of genetically modified organisms O.J.
    [Show full text]
  • Survival and Germination of Bacillus Clausii UBBC07 Spores in in Vitro Human Gastrointestinal Tract Simulation Model and Evaluation of Clausin Production
    fmicb-11-01010 June 8, 2020 Time: 20:24 # 1 ORIGINAL RESEARCH published: 10 June 2020 doi: 10.3389/fmicb.2020.01010 Survival and Germination of Bacillus clausii UBBC07 Spores in in vitro Human Gastrointestinal Tract Simulation Model and Evaluation of Clausin Production Jayesh J. Ahire*†, Megha S. Kashikar and Ratna Sudha Madempudi Centre for Research & Development, Unique Biotech Ltd., Hyderabad, India Bacillus clausii UBBC07 is a commercial spore probiotic known to reduce diarrhea Edited by: in children and adults. In the present study, survival and germination of UBBC07 Riadh Hammami, spores were investigated under fed and fasted conditions in Simulator of Human University of Ottawa, Canada Intestinal Microbial Ecosystem. Besides this, lantibiotic production, purification, and Reviewed by: characterization were performed. The agar plate analysis showed that spores were Emilia Ghelardi, University of Pisa, Italy 100% tolerant to fed and fasted gastrointestinal tract (GIT) conditions. Simultaneously, Sabrina Neves Casarotti, flow cytometry revealed that at the end of small intestinal incubation, 120% (fed) and Universidade Federal de Mato Grosso, Brazil 133% (fasted) spores were in viable germinating state. The transformation of viable Francesco Celandroni, germinating spores into viable vegetative cells was observed at 3 h of incubation under University of Pisa, Italy fasted GIT conditions. In antimicrobial evaluation, UBBC07 produced low-molecular- *Correspondence: weight (2107.94 Da) class I lantibiotic clausin. The presence of lanB, lanC, and lanD Jayesh J. Ahire [email protected]; genes confirms the clausin production. Clausin is stable at proteases (pepsin, proteinase [email protected] K, and trypsin), temperature (up to 100◦C), and pH (up to 11).
    [Show full text]
  • The Safety of Two Bacillus Probiotic Strains for Human Use
    Dig Dis Sci (2008) 53:954–963 DOI 10.1007/s10620-007-9959-1 ORIGINAL PAPER The Safety of Two Bacillus Probiotic Strains for Human Use Iryna B. Sorokulova Æ Iryna V. Pinchuk Æ Muriel Denayrolles Æ Irina G. Osipova Æ Jen M. Huang Æ Simon M. Cutting Æ Maria C. Urdaci Received: 11 April 2007 / Accepted: 1 August 2007 / Published online: 13 October 2007 Ó Springer Science+Business Media, LLC 2007 Abstract Probiotics based on Bacillus strains have been Introduction increasingly proposed for prophylactic and therapeutic use against several gastro-intestinal diseases. We studied safety Health promoting microorganisms, e.g., probiotics, have for two Bacillus strains included in a popular East Euro- been recently increasingly included in various food prod- pean probiotic. Bacillus subtilis strain that was sensitive ucts and proposed for use as a food supplement or as a to all antibiotics listed by the European Food Safety therapy for several infectious diseases [1, 2]. Probiotic Authority. Bacillus licheniformis strain was resistant to therapy is very attractive because it is an effective and non- chloramphenicol and clindamycin. Both were non-hemo- invasive low cost approach, which attempts to recreate lytic and did not produce Hbl or Nhe enterotoxins. No bceT natural flora rather than its disruption. Micro-organisms and cytK toxin genes were found. Study of acute toxicity used as a probiotic for human are mainly gram-positive in BALB/c mice demonstrated no treatment-related bacteria belonging to the Lactobacillus and Bacillus spp. deaths. The oral LD50 for both strains was more than For decades Bacillus bacteria and their metabolites have 2 · 1011 CFU.
    [Show full text]