Saccharomyces boulardii protease inhibits Clostridium difficile toxin A effects in the rat ileum

Castagliuolo, Ignazio; LaMont, J. Thomas; Nikulasson, S. T.; Pothoulakis, Charalabos

doi:10.1128/iai.64.12.5225-5232.1996

Cited by 219 publications

(118 citation statements)

References 38 publications

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“…reuteri RC14 can be used to improve the effectiveness of certain antibiotics in the treatment for Clostridium difficile secondary infection. While the exact mechanisms remain unknown, many hypotheses have been advanced to explain the antimicrobial activity of probiotics, notably competition for binding sites on the epithelium (Conway et al 1987), decreased intestinal cell permeability (Stratiki et al 2007), competition for nutrients (Sonnenburg et al 2006), degradation of toxin receptors (Castagliuolo et al 1996), stimulating the immune system (Kaila et al 1992) and production of various inhibitory substances such as organic acids (Vandenbergh 1993), bio-surfactants or hydrogen peroxide (Pridmore et al 2008), all previously reviewed (Fliss et al 2011). Secretion of cell signalling compounds such as reutericyclin (G€ anzle 2004) and bacteriocins has also been proposed as an antimicrobial mechanism.…”

Section: Introductionmentioning

confidence: 99%

Growth, acid production and bacteriocin production by probiotic candidates under simulated colonic conditions

et al. 2012

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Aims: The aim of this study is to evaluate the capacity of three bacteriocin producers, namely Lactococcus lactis subsp. lactis biovar diacetylactis UL719 (nisin Z producer), L. lactis ATCC 11454 (nisin A producer) and Pediococcus acidilactici UL5 (pediocin PA-1 producer), and to grow and produce their active bacteriocins in Macfarlane broth, which mimics the nutrient composition encountered in the human large intestine. Methods and Results:The three bacteriocin-producing strains were grown in Macfarlane broth and in De Man-Rogosa-Sharpe (MRS) broth. For each strain, the bacterial count, pH drop and production of organic acids and bacteriocins were measured for different period of time. The ability of the probiotic candidates to inhibit Listeria ivanovii HPB 28 in co-culture in Macfarlane broth was also examined. Lactococcus lactis subsp. lactis biovar diacetylactis UL719, L. lactis ATCC 11454 and Ped. acidilactici UL5 were able to grow and produce their bacteriocins in MRS broth and in Macfarlane broth. Each of the three candidates inhibited L. ivanovii HPB 28, and this inhibition activity was correlated with bacteriocin production. The role of bacteriocin production in the inhibition of L. ivanovii in Macfarlane broth was confirmed for Ped. acidilactici UL5 using a pediocin nonproducer mutant. Conclusions: The data provide some evidence that these bacteria can produce bacteriocins in a complex medium with carbon source similar to those found in the colon. Significance and Impact of the Study: This study demonstrates the capacity of lactic acid bacteria to produce their bacteriocins in a medium simulating the nutrient composition of the large intestine.

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Section: Introductionmentioning

confidence: 99%

Growth, acid production and bacteriocin production by probiotic candidates under simulated colonic conditions

et al. 2012

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“…Ileal secretion in response to Cl. difficile toxin A is inhibited by S. boulardii due to proteolysis of the toxin and inhibition of toxin binding to its receptor (Pothoulakis et al 1993;Castagliuolo et al 1996). Moreover, its preventive properties concerning traveller's diarrhoea (Kollaritsch et al 1993), antagonistic effects on Candida species (Ducluzeau and Bensaada 1982) as well as immunostimulatory effects (Caetano et al 1986;Buts et al 1990) have also been shown.…”

Section: Introductionmentioning

confidence: 99%

Clustering of Saccharomyces boulardii strains within the species S. cerevisiae using molecular typing techniques

Mitterdorfer

Mayer

Kneifel

et al. 2002

Journal of Applied Microbiology

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Aims: This study was undertaken to characterize and differentiate therapeutically relevant Saccharomyces yeasts. Among the isolates were so-called Saccharomyces boulardii strains, which are considered as probiotic agents, but whose taxonomic assignment is controversial. Moreover, the discriminative power of the applied molecular typing techniques should be evaluated. Methods and Results: Genotyping was performed using species-specific polymerase chain reaction (PCR), randomly amplified polymorphic DNA-PCR, restriction fragment length polymorphism analysis of rDNA spacer regions and pulsed-field gel electrophoresis. Speciesspecific PCR assigned all of the product isolates to the species S. cerevisiae. By combining the other techniques, all isolates could be discriminated. Moreover, it could be demonstrated that probiotic S. boulardii strains form a separate cluster located within the species. Conclusions: With the exception of species-specific PCR, all of the applied methodologies were suitable for subspecies typing and indicated a close relationship between the probiotic strains. Significance and Impact of the Study: The methods applied in this study are considered powerful tools for quality control of therapeutically relevant yeasts. It is of crucial importance, especially regarding S. boulardii yeasts, to verify the identity of the correct strain, since the beneficial properties are considered to be strain-specific.

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“…Despite one died after the C. difficile challenge, the diarrhea symptoms were alleviated in the remaining animals, and a few became normal after re-administration of L. lactis carrying the empty plasmid. Together these observations indicate that the probiotic L. lactis has a protective effect on CDI animal model and may prevent C. difficile-associated diarrhea in animals by inhibiting C. difficile colonization and/or other mechanisms [18,19].…”

Section: Effect Of Recombinant L Lactis Live Vaccine On C Difficilementioning

confidence: 71%

The protective effect of recombinant Lactococcus lactis oral vaccine on a Clostridium difficile-infected animal model

et al. 2013

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BackgroundOral immunization with vaccines may be an effective strategy for prevention of Clostridium difficile infection (CDI). However, application of previously developed vaccines for preventing CDI has been limited due to various reasons. Here, we developed a recombinant Lactococcus lactis oral vaccine and evaluated its effect on a C. difficile-infected animal model established in golden hamsters in attempt to provide an alternative strategy for CDI prevention.MethodsRecombinant L. lactis vaccine was developed using the pTRKH2 plasmid, a high-copy-number Escherichia coli-L. shuttle vector: 1) L. lactis expressing secreted proteins was constructed with recombinant pTRKH2 (secreted-protein plasmid) carrying the Usp45 signal peptide (SPUsp45), nontoxic adjuvanted tetanus toxin fragment C (TETC), and 14 of the 38 C-terminal repeats (14CDTA) of nontoxic C. difficile toxin A (TcdA); and 2) L. lactis expressing secreted and membrane proteins was constructed with recombinant pTRKH2 (membrane-anchored plasmid) carrying SPUsp45, TETC, 14CDTA, and the cell wall-anchored sequence of protein M6 (cwaM6). Then, 32 male Syrian golden hamsters were randomly divided into 4 groups (n = 8 each) for gavage of normal saline (blank control) and L. lactis carrying the empty shuttle vector, secreted-protein plasmid, and membrane-anchored plasmid, respectively. After 1-week gavage of clindamycin, the animals were administered with C. difficile spore suspension. General symptoms and intestinal pathological changes of the animals were examined by naked eye and microscopy, respectively. Protein levels of anti-TcdA IgG/IgA antibodies in intestinal tissue and fluid were analyzed by enzyme-linked immunosorbent assay (ELISA). A cell culture cytotoxicity neutralization assay was done by TcdA treatment with or without anti-TcdA serum pre-incubation or treatment. Apoptosis of intestinal epithelial cells was examined by flow cytometry (FL) assay. Expression of mucosal inflammatory cytokines in the animals was detected by polymer chain reaction (PCR) assay.ResultsAfter the C. difficile challenge, the animals of control group had severe diarrhea symptoms on day 1 and all died on day 4, indicating that the CDI animal model was established in hamster. Of the 3 immunization groups, secreted-protein and membrane-anchored plasmid groups had significantly lower mortalities, body weight decreases, and pathological scores, with higher survival rate/time than the empty plasmid group (P < 0.05). The tilter of IgG antibody directed against TcdA was significantly higher in serum and intestinal fluid of secreted-protein and membrane-anchored plasmid groups than in the empty plasmid group (P < 0.05) while the corresponding titer of IgA antibody directed against TcdA had no substantial differences (P > 0.05). The anti-TcdA serum of membrane-anchored plasmid group neutralized the cytotoxicity of 200 ng/ml TcdA with the best protective effect achieved by anti-TcdA serum pre-incubation. The incidences of TcdA-induced death and apoptosis of intestinal epithelial cell...

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Saccharomyces boulardii protease inhibits Clostridium difficile toxin A effects in the rat ileum

Cited by 219 publications

References 38 publications

Growth, acid production and bacteriocin production by probiotic candidates under simulated colonic conditions

Growth, acid production and bacteriocin production by probiotic candidates under simulated colonic conditions

Clustering of Saccharomyces boulardii strains within the species S. cerevisiae using molecular typing techniques

The protective effect of recombinant Lactococcus lactis oral vaccine on a Clostridium difficile-infected animal model

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