Effect of adding sodium taurocholate to selective media on the recovery of Clostridium difficile from environmental surfaces.

ABSTRACTRecurrentClostridium difficileinfection (R-CDI) is common and difficult to treat, potentially necessitating fecal microbiota transplantation (FMT). AlthoughC. difficilespores persist in the hospital environment and cause infection, little is known about their potential presence or importance in the household environment. Households of R-CDI subjects in the peri-FMT period and of geographically matched and age-matched controls were analyzed for the presence ofC. difficile. Household environmental surfaces and fecal samples from humans and pets in the household were examined. Households of post-FMT subjects were also examined (environmental surfaces only). Participants were surveyed regarding their personal history and household cleaning habits. Species identity and molecular characteristics of presumptiveC. difficileisolates from environmental and fecal samples were determined by using the Pro kit (Remel, USA), Gram staining, PCR, toxinotyping,tcdCgene sequencing, and pulsed-field gel electrophoresis (PFGE). Environmental cultures detectedC. difficileon ≥1 surface in 8/8 (100%) peri-FMT households, versus 3/8 (38%) post-FMT households and 3/8 (38%) control households (P= 0.025). The most commonC. difficile-positive sites were the vacuum (11/27; 41%), toilet (8/30; 27%), and bathroom sink (5/29; 17%).C. difficilewas detected in 3/36 (8%) fecal samples (two R-CDI subjects and one household member). Nine (90%) of 10 households with multipleC. difficile-positive samples had a single genotype present each. In conclusion,C. difficilewas found in the household environment of R-CDI patients, but whether it was found as a cause or consequence of R-CDI is unknown. If household contamination leads to R-CDI, effective decontamination may be protective.

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Germination efficiency of clinical Clostridium difficile spores and correlation with ribotype, disease severity and therapy failure

Journal of Medical Microbiology ◽

10.1099/jmm.0.056614-0 ◽

2013 ◽

Vol 62 (9) ◽

pp. 1405-1413 ◽

Cited By ~ 29

Author(s):

P. Moore ◽

L. Kyne ◽

A. Martin ◽

K. Solomon

Keyword(s):

Clostridium Difficile ◽

Spore Germination ◽

Germination Rate ◽

Sodium Taurocholate ◽

Response To Treatment ◽

First Line Therapy ◽

High Germination ◽

Forming Efficiency ◽

Spore Outgrowth ◽

Ribotype 078

Spore germination is an important part of the pathogenesis of Clostridium difficile infection (CDI). Spores are resistant to antibiotics, including those therapeutically administered for CDI and strains with a high germination rate are significantly more likely to be implicated in recurrent CDI. The role of germination efficiency in cases of refractory CDI where first-line therapy fails remains unclear. We investigated spore germination efficiencies of clinical C. difficile isolates by measuring drop in OD600 and colony forming efficiency. Ribotype 027 isolates exhibited significantly higher germination efficiencies in the presence of 0.1 % (w/v) sodium taurocholate (51.66±8.75 %; 95 % confidence interval (CI) 47.37–55.95 %) than ribotype 106 (41.91±8.35 %; 95 % CI 37.82–46 %) (P<0.05) and ribotype 078 (42.07±8.57 %, 95 % CI 37.22–46.92 %) (P<0.05). Spore outgrowth rates were comparable between the ribotype groups but the exponential phase occurred approximately 4 h later in the absence of sodium taurocholate. Spore germination efficiencies for isolates implicated in severe CDI were significantly higher (49.68±10.00 %, 95 % CI 47.06–52.30 %) than non-severe CDI (40.92±9.29 %, 95 % CI 37.48–44.36 %); P<0.01. Germination efficiencies were also significantly higher in recurrent CDI or when metronidazole therapy failed than when therapy was successful [(49.00±10.49 %, 95 % CI 46.25–51.75 %) versus (41.42±9.43 %, 95 % CI 37.93–44.91 %); P<0.01]. This study suggests an important link between C. difficile spore germination, CDI pathogenesis and response to treatment; however, further work is warranted before the complex interplay between germination dynamics and CDI outcome can be fully understood.

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Isolation of Environmental Clostridium Difficile from a Veterinary Teaching Hospital

Journal of Veterinary Diagnostic Investigation ◽

10.1177/104063870001200510 ◽

2000 ◽

Vol 12 (5) ◽

pp. 449-452 ◽

Cited By ~ 40

Author(s):

J. Scott Weese ◽

Henri R. Staempfli ◽

John F. Prescott

Keyword(s):

Clostridium Difficile ◽

Teaching Hospital ◽

Potential Role ◽

Sodium Taurocholate ◽

Fecal Contamination ◽

Small Animal ◽

Large Animal ◽

High Animal ◽

Nosocomial Disease

An environmental survey of a veterinary teaching hospital for the presence of Clostridium difficile was performed using contact plates and cycloserine-cefoxitin-fructose with 0.1% sodium taurocholate agar. Clostridium difficile was isolated from 24 of 381 sites (6.3%). Growth was obtained from 4.5% (9/202) of sites sampled in the Large Animal Clinic, from 8.1% (13/160) of sites within the Small Animal Clinic, and from 20% (2/10) of sites sampled elsewhere. Fourteen of 21 strains tested produced toxins in vitro. A geographic association was found with areas in the large animal clinic where nosocomial C. difficile diarrhea in horses had previously been diagnosed. Several other sites with a potential for nosocomial transmission of the organism were identified. Areas from which C. difficile was isolated tended to be areas with high animal traffic, with increased chance of fecal contamination, and with rough, difficult to clean surfaces. This study documents the prevalence of this organism in the environment and its potential role in nosocomial disease.

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Use of sodium taurocholate to enhance spore recovery on a medium selective for Clostridium difficile.

Journal of Clinical Microbiology ◽

10.1128/jcm.15.3.443-446.1982 ◽

1982 ◽

Vol 15 (3) ◽

pp. 443-446 ◽

Cited By ~ 189

Author(s):

K H Wilson ◽

M J Kennedy ◽

F R Fekety

Keyword(s):

Clostridium Difficile ◽

Sodium Taurocholate

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Cold-Air Atmospheric Pressure Plasma Against Clostridium difficile Spores: A Potential Alternative for the Decontamination of Hospital Inanimate Surfaces

Infection Control and Hospital Epidemiology ◽

10.1017/ice.2015.39 ◽

2015 ◽

Vol 36 (6) ◽

pp. 742-744 ◽

Cited By ~ 9

Author(s):

Tânia Claro ◽

Orla J. Cahill ◽

Niall O’Connor ◽

Stephen Daniels ◽

Hilary Humphreys

Keyword(s):

Clostridium Difficile ◽

Atmospheric Pressure ◽

Atmospheric Pressure Plasma ◽

Infect Control Hosp ◽

Air Plasma ◽

Cold Air ◽

Pressure Plasma ◽

Environmental Surfaces ◽

Potential Alternative ◽

Single Jet

AbstractClostridium difficile spores survive for months on environmental surfaces and are highly resistant to decontamination. We evaluated the effect of cold-air plasma against C. difficile spores. The single-jet had no effect while the multi-jet achieved 2–3 log10 reductions in spore counts and may augment traditional decontamination.Infect Control Hosp Epidemiol 2015;00(0):1–3

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Isolation of Clostridium difficile from faecal specimens – a comparison of chromID C. difficile agar and cycloserine-cefoxitin-fructose agar

Journal of Medical Microbiology ◽

10.1099/jmm.0.056515-0 ◽

2013 ◽

Vol 62 (9) ◽

pp. 1423-1427 ◽

Cited By ~ 22

Author(s):

Kerry C. Carson ◽

Lusiana V. Boseiwaqa ◽

Sara K. Thean ◽

Niki F. Foster ◽

Thomas V. Riley

Keyword(s):

Clostridium Difficile ◽

Sodium Taurocholate ◽

Laboratory Diagnosis ◽

Strain Typing ◽

Isolation And Identification ◽

Time Saving ◽

Faecal Samples ◽

Significant Difference ◽

Chromogenic Agar ◽

Stool Specimens

The culture of toxigenic Clostridium difficile from stool specimens is still seen as the gold standard for the laboratory diagnosis of C. difficile infection (CDI). bioMérieux have released ChromID Cdiff chromogenic agar (CDIF) for the isolation and identification of C. difficile in 24 h. In this study, we compared CDIF to pre-reduced cycloserine-cefoxitin-fructose agar with sodium taurocholate (TCCFA) in the examination of glutamate dehydrogenase-positive faecal specimens that were either GeneOhm positive or negative, using direct culture or culture following alcohol shock. Direct culture on CDIF had a sensitivity of 100 % and recovery of 94 % while for TCCFA these were 87 % and 82 %, respectively. For GeneOhm-positive alcohol-shocked faecal samples, sensitivity and recovery on CDIF was similar to direct culture while on TCCFA they were about 10 % higher. For direct culture, there was a significant difference between growth on CDIF at 24 h and TCCFA at 48 h (P = 0.001) and between the two media at 48 h (P<0.001). A total of 142 strains of C. difficile were recovered in pure culture from all GeneOhm-positive samples used in this study and 11 (7.7 %) of these were A−B−CDT− and may represent mixed infections of toxigenic and non-toxigenic C. difficile. The most dominant ribotype was UK 014 (14.7 %) followed by 002 (11.9 %) and 020 (11.9 %), and 36 % of toxigenic isolates, including an A−B+CDT− strain, could not be assigned a UK ribotype. CDIF outperformed pre-reduced TCCFA by negating the need for alcohol shock treatment and by giving a time saving of 24 h in the isolation of C. difficile. CDIF plates were also more selective than TCCFA and C. difficile colonies were easy to identify and subculture prior to strain typing.

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Clostridium difficile from Healthy Food Animals: Optimized Isolation and Prevalence†

Journal of Food Protection ◽

10.4315/0362-028x.jfp-10-229 ◽

2011 ◽

Vol 74 (1) ◽

pp. 130-133 ◽

Cited By ~ 21

Author(s):

S. N. THITARAM ◽

J. F. FRANK ◽

S. A. LYON ◽

G. R. SIRAGUSA ◽

J. S. BAILEY ◽

...

Keyword(s):

Clostridium Difficile ◽

Beef Cattle ◽

Dairy Cattle ◽

Sodium Taurocholate ◽

Fecal Samples ◽

Selective Enrichment ◽

Sheep Blood ◽

Cattle Feces ◽

Isolation Methods ◽

Better Than

Two isolation methods were compared for isolation of Clostridium difficile from food animal feces. The single alcohol shock method (SS) used selective enrichment in cycloserine-cefoxitin fructose broth supplemented with 0.1% sodium taurocholate, followed by alcohol shock and isolation on tryptic soy agar supplemented with 5% sheep blood, and cycloserine-cefoxitin fructose agar. The double alcohol shock method (DS) used alcohol shock prior to and after selective enrichment in cycloserinecefoxitin fructose broth supplemented with 0.1% sodium taurocholate, followed by isolation on tryptic soy agar supplemented with 5% sheep blood and cycloserine-cefoxitin fructose agar. A total of 55 (15.9%, n = 345) swine fecal samples, 32 (2.4%, n = 1,325) dairy cattle fecal samples, and 188 (6.3%, n = 2,965) beef cattle fecal samples were positive for C. difficile by either method. However, the DS was significantly better than the SS for the recovery of C. difficile from swine feces, while the SS was significantly better than the DS for the recovery of C. difficile from beef cattle feces. There was no significant difference between methods for the recovery of C. difficile from dairy cattle feces. This study suggests that food animals might harbor C. difficile and it provides critical information that isolation methods might not have universal application across animal species.

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