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Association of Fidaxomicin with C. difficile Spores: Effects of Persistence on Subsequent Spore Recovery, Outgrowth and Toxin Production.
PLoS One. 2016; 11(8):e0161200.Plos

Abstract

BACKGROUND

We have previously shown that fidaxomicin instillation prevents spore recovery in an in-vitro gut model, whereas vancomycin does not. The reasons for this are unclear. Here, we have investigated persistence of fidaxomicin and vancomycin on C. difficile spores, and examined post-antibiotic exposure spore recovery, outgrowth and toxin production.

METHODS

Prevalent UK C. difficile ribotypes (n = 10) were incubated with 200mg/L fidaxomicin, vancomycin or a non-antimicrobial containing control for 1 h in faecal filtrate or Phosphate Buffered Saline. Spores were washed three times with faecal filtrate or phosphate buffered saline, and residual spore-associated antimicrobial activity was determined by bioassay. For three ribotypes (027, 078, 015), antimicrobial-exposed, faecal filtrate-washed spores and controls were inoculated into broth. Viable vegetative and spore counts were enumerated on CCEYL agar. Percentage phase bright spores, phase dark spores and vegetative cells were enumerated by phase contrast microscopy at 0, 3, 6, 24 and 48 h post-inoculation. Toxin levels (24 and 48h) were determined by cell cytotoxicity assay.

RESULTS

Fidaxomicin, but not vancomycin persisted on spores of all ribotypes following washing in saline (mean = 10.1mg/L; range = 4.0-14mg/L) and faecal filtrate (mean = 17.4mg/L; 8.4-22.1mg/L). Outgrowth and proliferation rates of vancomycin-exposed spores were similar to controls, whereas fidaxomicin-exposed spores showed no vegetative cell growth after 24 and 48 h. At 48h, toxin levels averaged 3.7 and 3.3 relative units (RU) in control and vancomycin-exposed samples, respectively, but were undetectable in fidaxomicin-exposed samples.

CONCLUSION

Fidaxomicin persists on C. difficile spores, whereas vancomycin does not. This persistence prevents subsequent growth and toxin production in vitro. This may have implications on spore viability, thereby impacting CDI recurrence and transmission rates.

Authors+Show Affiliations

Leeds Institute for Biomedical and Clinical Sciences, University of Leeds, Leeds, United Kingdom.Faculty of Science and Engineering, Manchester Metropolitan University, Manchester, United Kingdom.Leeds Institute for Biomedical and Clinical Sciences, University of Leeds, Leeds, United Kingdom.Astellas Pharma Europe Ltd, Chertsey, Surrey, United Kingdom.Leeds Institute for Biomedical and Clinical Sciences, University of Leeds, Leeds, United Kingdom. Department of Microbiology, Leeds Teaching Hospitals NHS Trust, The General Infirmary, Old Medical School, Leeds, United Kingdom.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

27556739

Citation

Chilton, Caroline H., et al. "Association of Fidaxomicin With C. Difficile Spores: Effects of Persistence On Subsequent Spore Recovery, Outgrowth and Toxin Production." PloS One, vol. 11, no. 8, 2016, pp. e0161200.
Chilton CH, Crowther GS, Ashwin H, et al. Association of Fidaxomicin with C. difficile Spores: Effects of Persistence on Subsequent Spore Recovery, Outgrowth and Toxin Production. PLoS ONE. 2016;11(8):e0161200.
Chilton, C. H., Crowther, G. S., Ashwin, H., Longshaw, C. M., & Wilcox, M. H. (2016). Association of Fidaxomicin with C. difficile Spores: Effects of Persistence on Subsequent Spore Recovery, Outgrowth and Toxin Production. PloS One, 11(8), e0161200. https://doi.org/10.1371/journal.pone.0161200
Chilton CH, et al. Association of Fidaxomicin With C. Difficile Spores: Effects of Persistence On Subsequent Spore Recovery, Outgrowth and Toxin Production. PLoS ONE. 2016;11(8):e0161200. PubMed PMID: 27556739.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Association of Fidaxomicin with C. difficile Spores: Effects of Persistence on Subsequent Spore Recovery, Outgrowth and Toxin Production. AU - Chilton,Caroline H, AU - Crowther,Grace S, AU - Ashwin,Helen, AU - Longshaw,Chris M, AU - Wilcox,Mark H, Y1 - 2016/08/24/ PY - 2016/06/27/received PY - 2016/08/01/accepted PY - 2016/8/25/entrez PY - 2016/8/25/pubmed PY - 2017/7/29/medline SP - e0161200 EP - e0161200 JF - PloS one JO - PLoS ONE VL - 11 IS - 8 N2 - BACKGROUND: We have previously shown that fidaxomicin instillation prevents spore recovery in an in-vitro gut model, whereas vancomycin does not. The reasons for this are unclear. Here, we have investigated persistence of fidaxomicin and vancomycin on C. difficile spores, and examined post-antibiotic exposure spore recovery, outgrowth and toxin production. METHODS: Prevalent UK C. difficile ribotypes (n = 10) were incubated with 200mg/L fidaxomicin, vancomycin or a non-antimicrobial containing control for 1 h in faecal filtrate or Phosphate Buffered Saline. Spores were washed three times with faecal filtrate or phosphate buffered saline, and residual spore-associated antimicrobial activity was determined by bioassay. For three ribotypes (027, 078, 015), antimicrobial-exposed, faecal filtrate-washed spores and controls were inoculated into broth. Viable vegetative and spore counts were enumerated on CCEYL agar. Percentage phase bright spores, phase dark spores and vegetative cells were enumerated by phase contrast microscopy at 0, 3, 6, 24 and 48 h post-inoculation. Toxin levels (24 and 48h) were determined by cell cytotoxicity assay. RESULTS: Fidaxomicin, but not vancomycin persisted on spores of all ribotypes following washing in saline (mean = 10.1mg/L; range = 4.0-14mg/L) and faecal filtrate (mean = 17.4mg/L; 8.4-22.1mg/L). Outgrowth and proliferation rates of vancomycin-exposed spores were similar to controls, whereas fidaxomicin-exposed spores showed no vegetative cell growth after 24 and 48 h. At 48h, toxin levels averaged 3.7 and 3.3 relative units (RU) in control and vancomycin-exposed samples, respectively, but were undetectable in fidaxomicin-exposed samples. CONCLUSION: Fidaxomicin persists on C. difficile spores, whereas vancomycin does not. This persistence prevents subsequent growth and toxin production in vitro. This may have implications on spore viability, thereby impacting CDI recurrence and transmission rates. SN - 1932-6203 UR - https://www.unboundmedicine.com/medline/citation/27556739/Association_of_Fidaxomicin_with_C__difficile_Spores:_Effects_of_Persistence_on_Subsequent_Spore_Recovery_Outgrowth_and_Toxin_Production_ L2 - http://dx.plos.org/10.1371/journal.pone.0161200 DB - PRIME DP - Unbound Medicine ER -