Effects of Postbiotic from Bacteriocin-Like Inhibitory Substance Producing Enterococcus faecalis on Toxigenic Clostridioides difficile
Abstract
Objective: To determine the activities of postbiotics, prepared from bacteriocin-like producing Enterococcus faecalis (E. faecalis) against Clostridioides difficile (C. difficile) and its spores, and to assess the safety of postbiotics in vivo.
Material and Methods: Bacteriocin production of E. faecalis PK1201 was screened by proteolytic enzyme treatment, and bacteriocin-encoding genes were characterized by whole genome sequencing. A postbiotic of E. faecalis PK1201 was prepared using neutralized cell-free supernatant or bacterial cell lysate, which was then used to screen for antimicrobial activity via agar well diffusion. The lyophilized cell-free supernatant (LCFS) was further determined for its minimum inhibitory concentration (MIC) against C. difficile 630. The morphological changes of C. difficile were observed under scanning electron microscopes (SEM). Subsequently, the LCFS at sub-MIC and MIC were used to evaluate anti-spore germination activity. Finally, the safety of postbiotics was accessed using the Galleria mellonella model.
Results: E. faecalis PK1201 carried enterolysin A encoding gene. For postbiotic preparation, only LCFS exhibited antimicrobial activity, and the activity was completely lost after proteinase K treatment; indicating the existence of bacteriocin. LCFS showed anti-C. difficile with MIC of 18.2±6.9 mg/mL. The SEM images demonstrated shorter destruction of C. difficile cells after being treated with LCFS. Interestingly, LCFS at the MIC and sub-MIC revealed anti-spore germination activity against toxigenic C. difficile compared to the control. LCFS showed no acute toxicity in G. mellonella at the tested concentration.
Conclusion: The powerful activity and safety of LCFS shed light on the role of postbiotics in pharmaceutical products to control CDI.
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Centers for Disease Control and Prevention. C. diff (Clostridioides difficile) [homepage on the Internet]. Georgia: Centers for Disease Control and Prevention 2015 [cite 2022 Jul 1]. Available from: https://www.cdc.gov/cdiff/index.html
Kelly CP, LaMont JT. Clostridium difficile infection. Annu Rev Med 1998;49:375-90.
Abou Chakra CN, Pepin J, Sirard S, Valiquette L. Risk factors for recurrence, complications and mortality in Clostridium difficile infection: a systematic review. PLoS One 2014;9:e98400.
FAO/WHO. Evaluation of health and nutritional properties of powder milk and live lactic acid bacteria. Cordoba: Report of Joint FAO/WHO Expert Consultation; 2001;p.1-4.
O’sullivan L, Ross RP, Hill C. Potential of bacteriocin-producing lactic acid bacteria for improvements in food safety and quality. Biochimie 2002;84:593-604.
Delves-Broughton J, Blackburn P, Evans RJ, Hugenholtz J. Applications of the bacteriocin, nisin. Antonie Van Leeuwenhoek 1996;69:193-202.
Cohen SH, Gerding DN, Johnson S, Kelly CP, Loo VG, McDonald LC, et al. Clinical practice guidelines for Clostridium difficile infection in adults: 2010 update by the Society for Healthcare Epidemiology of America (SHEA) and the Infectious Diseases Society of America (IDSA). Infec Control Hosp Epidemiol 2010;31:431-55.
Parkes GC, Sanderson JD, Whelan K. The mechanisms and efficacy of probiotics in the prevention of Clostridium difficileassociated diarrhoea. Lancet Infect Dis 2009;9:237-44.
Mansour NM, Elkhatib WF, Aboshanab KM, Bahr M. Inhibition of Clostridium difficile in mice using a mixture of potential probiotic strains Enterococcus faecalis NM815, E. faecalis NM915, and E. faecium NM1015: novel candidates to control C. difficile infection (CDI). Probiotics Antimicro Prot 2018;10:511-22.
Romyasamit C, Thatrimontrichai A, Aroonkesorn A, Chanket W, Ingviya N, Saengsuwan P, et al. Enterococcus faecalis Isolated From Infant Feces Inhibits Toxigenic Clostridioides (Clostridium) difficile. Front Pedia 2020:612.
Doron S, Snydman DR. Risk and safety of probiotics. Clin Infect Dis 2015;60(Suppl_2):S129-34.
Salminen S, Collado MC, Endo A, Hill C, Lebeer S, Quigley EM, et al. The International Scientific Association of Probiotics and Prebiotics (ISAPP) consensus statement on the definition and scope of postbiotics. Nat Rev Gastroenterol Hepatol 2021;18: 649-67.
Šušković J, Kos B, Beganović J, Leboš Pavunc A, Habjanič K, Matošić S. Antimicrobial activity–the most important property of probiotic and starter lactic acid bacteria. Food Technol Biotechnol 2010;48:296-307.
De Almeida Júnior WL, da Silva Ferrari Í, de Souza JV, da Silva CD, da Costa MM, Dias FS. Characterization and evaluation of lactic acid bacteria isolated from goat milk. Food Control 2015;53:96-103.
Aguilar-Toalá J, Garcia-Varela R, Garcia H, Mata-Haro V, González-Córdova A, Vallejo-Cordoba B, et al. Postbiotics: an evolving term within the functional foods field. Trends Food Sci Technol 2018;75:105-14.
Chu TW, Chen CN, Pan CY. Antimicrobial status of tilapia (Oreochromis niloticus) fed Enterococcus adium originally isolated from goldfish intestine. Aquac Rep 2020;17:100397.
Breukink E, van Heusden HE, Vollmerhaus PJ, Swiezewska E, Brunner L, Walker S, et al. Lipid II is an intrinsic component of the pore induced by nisin in bacterial membranes. J Biol Chem 2003;278:19898-903.
Redondo LM, Carrasco JMD, Redondo EA, Delgado F, Miyakawa MEF. Clostridium perfringens type E virulence traits involved in gut colonization. PloS One 2015;10:e0121305.
Han S-K, Shin M-S, Park H-E, Kim S-Y, Lee W-K. Screening of bacteriocin-producing Enterococcus faecalis strains for antagonistic activities against Clostridium perfringens. J Food Sci Anim Resour 2014;34:614.
Bolger AM, Lohse M, Usadel B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 2014;30:2114-20.
Bankevich A, Nurk S, Antipov D, Gurevich AA, Dvorkin M, Kulikov AS, et al. SPAdes: a new genome assembly algorithm and its applications to single-cell sequencing. J Comput Biol 2012;19:455-77.
Gurevich A, Saveliev V, Vyahhi N, Tesler G. QUAST: quality assessment tool for genome assemblies. Bioinformatics 2013; 29:1072-5.
Seemann T. Prokka: rapid prokaryotic genome annotation. Bioinformatics 2014;30:2068-9.
van Heel AJ, de Jong A, Song C, Viel JH, Kok J, Kuipers OP. BAGEL4: a user-friendly web server to thoroughly mine RiPPs and bacteriocins. Nucleic Acids Res 2018;46(W1):W278-81.
Shin HS, Park SY, Lee DK, Kim S, An HM, Kim JR, et al. Hypocholesterolemic effect of sonication-killed Bifidobacterium longum isolated from healthy adult Koreans in high cholesterol fed rats. Arch Pharmacal Res 2010;33:1425-31.
Lynch T, Chong P, Zhang J, Hizon R, Du T, Graham MR, et al. Characterization of a stable, metronidazole-resistant Clostridium difficile clinical isolate. PloS One 2013;8:e53757.
Lawley TD, Croucher NJ, Yu L, Clare S, Sebaihia M, Goulding D, et al. Proteomic and genomic characterization of highly infectious Clostridium difficile 630 spores. J Bacteriol 2009;191:5377-86.
Carlson Jr PE, Kaiser AM, McColm SA, Bauer JM, Young VB, Aronoff DM, et al. Variation in germination of Clostridium difficile clinical isolates correlates to disease severity. Anaerobe 2015;33:64-70.
Rossoni RD, De Barros PP, Mendonça IdC, Medina RP, Silva DHS, Fuchs BB, et al. The postbiotic activity of Lactobacillus paracasei 28.4 against Candida auris. Front Cell Infect Microbiol 2020:397. doi: 10.3389/fcimb.2020.00397
Vincent C, Manges AR. Antimicrobial use, human gut microbiota and Clostridium difficile colonization and infection. Antibiotics 2015;4:230-53.
Morniroli D, Vizzari G, Consales A, Mosca F, Giannì ML. Postbiotic supplementation for children and newborn’s health. Nutrients 2021;13:781.
Teame T, Wang A, Xie M, Zhang Z, Yang Y, Ding Q, et al. Paraprobiotics and postbiotics of probiotic Lactobacilli, their positive effects on the host and action mechanisms: A review. Front Nutr 2020:191.
Caly DL, Chevalier M, Flahaut C, Cudennec B, Al Atya AK, Chataigné G, et al. The safe enterocin DD14 is a leaderless two-peptide bacteriocin with anti-Clostridium perfringens activity. Int J Antimicrob Agents 2017;49:282-9.
Wu Y, Pang X, Wu Y, Liu X, Zhang X. Enterocins: classification, synthesis, antibacterial mechanisms and food applications. Molecules 2022;27:2258.
Nilsen T, Nes IF, Holo H. Enterolysin_A, a cell wall-degrading bacteriocin from Enterococcus faecalis LMG 2333. Appl Environ Microbiol 2003;69:2975-84.
Zhu D, Sorg JA, Sun X. Clostridioides difficile biology: sporulation, germination, and corresponding therapies for C. difficile infection. Front Cell Infect Microbio 2018;8:29.
Thanissery R, Winston JA, Theriot CM. Inhibition of spore germination, growth, and toxin activity of clinically relevant C. difficile strains by gut microbiota derived secondary bile acids. Anaerobe 2017;45:86-100.
Bourgin M, Kriaa A, Mkaouar H, Mariaule V, Jablaoui A, Maguin E, et al. Bile Salt Hydrolases: At the crossroads of microbiota and human health. Microorganisms 2021;9:1122.
Thanissery R, Winston JA, Theriot CM. Inhibition of spore germination, growth, and toxin activity of clinically relevant C. difficile strains by gut microbiota derived secondary bile acids. Anaerobe 2017;45:86-100.
Thanissery R, Winston JA, Theriot CM. Inhibition of spore germination, growth, and toxin activity of clinically relevant C. difficile strains by gut microbiota derived secondary bile acids. Anaerobe 2017;45:86-100.
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