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Co-culture with potentially probiotic microorganisms antagonises virulence factors of Clostridium difficile in vitro

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Abstract

Toxigenic strains of Clostridium difficile were co-cultured with different strains of bifidobacteria and lactobacilli. Spent culture supernatants were tested for biological activity on cultured Vero cells. Co-culture of C. difficile with some potentially probiotic strains lead to a reduction of the biological activity of spent culture supernatants. The observed effects cannot be ascribed either to secreted factors from the probiotic strains or to toxin adsorption by bacterial cells. Immunological assays showed that there was significant diminution of both clostridial toxins (TcdA and TcdB) in spent culture supernatants of co-cultures as compared with pure clostridial cultures. Even though co-cultured clostridial cells showed a slight increase of intracellular toxins, this increase did not completely explains the reduction of toxin concentration in culture supernatants. The evidence suggests that the antagonism could be due to the diminution of the synthesis and/or secretion of both clostridial toxins. Our findings provide new insights into the possible mechanisms involved in the protective effect of probiotics in the context of C. difficile infection.

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Abbreviations

SCS:

Spent culture supernatants

TcdA:

C. difficile toxin A

TcdB:

C. difficile toxin B

CDAD:

C. difficile associated diarrhea

OD600 nm :

Optical density at 600 nm

PBS:

Phosphate buffered saline

DMEM:

Dulbbeco’s Modified Eagle’s Medium

rd:

Ratio of detached cells

NBT:

Nitro-blue tetrazolium chloride

BCIP:

5-Bromo-4-chloro-3′-indolyphosphate p-toluidine salt

DD50:

Dose of SCS that leads to the detachment of 50% of the cells

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Authors and Affiliations

  1. CCT La Plata, CONICET, Centro de Investigación y Desarrollo en Criotecnología de Alimentos (CIDCA), Facultad de Ciencias Exactas, Universidad Nacional de La Plata, 47 y 116 (1900), La Plata, Argentina

    Fernando M. Trejo, Pablo F. Pérez & Graciela L. De Antoni

  2. Cátedra de Microbiología, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Argentina

    Pablo F. Pérez & Graciela L. De Antoni

  3. CIC-PBA, Buenos Aires, Argentina

    Graciela L. De Antoni

Authors
  1. Fernando M. Trejo
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  2. Pablo F. Pérez
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  3. Graciela L. De Antoni
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Correspondence to Pablo F. Pérez.

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Trejo, F.M., Pérez, P.F. & De Antoni, G.L. Co-culture with potentially probiotic microorganisms antagonises virulence factors of Clostridium difficile in vitro. Antonie van Leeuwenhoek 98, 19–29 (2010). https://doi.org/10.1007/s10482-010-9424-6

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  • DOI: https://doi.org/10.1007/s10482-010-9424-6

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