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

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

Journal of Clinical Microbiology ◽

10.1128/jcm.21.4.636-637.1985 ◽

1985 ◽

Vol 21 (4) ◽

pp. 636-637 ◽

Cited By ~ 17

Author(s):

B P Buggy ◽

C C Hawkins ◽

R Fekety

Keyword(s):

Clostridium Difficile ◽

Sodium Taurocholate ◽

Selective Media ◽

Environmental Surfaces

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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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Sporulation of Clostridium difficile in Aerobic conditions is Significantly Protracted when Exposed to Sodium Taurocholate

Journal of Medical Microbiology & Diagnosis ◽

10.4172/2161-0703.1000230 ◽

2016 ◽

Vol 05 (02) ◽

Author(s):

Tony Worthington ◽

Anthony C Hilton

Keyword(s):

Clostridium Difficile ◽

Sodium Taurocholate ◽

Aerobic Conditions

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First molecular characterisation and PCR ribotyping of Clostridium difficile strains isolated in two Algerian Hospitals

The Journal of Infection in Developing Countries ◽

10.3855/jidc.9580 ◽

2018 ◽

Vol 12 (01) ◽

pp. 015-021 ◽

Cited By ~ 3

Author(s):

Abla Djebbar ◽

Mohammed Sebaihia ◽

Ed Kuijper ◽

Celine Harmanus ◽

Ingrid Sanders ◽

...

Keyword(s):

Health Care ◽

Clostridium Difficile ◽

Sodium Taurocholate ◽

Health Care Facilities ◽

Hospitalized Patients ◽

Test Method ◽

Surveillance Program ◽

Local Health ◽

Pcr Ribotyping ◽

Considerable Morbidity

Introduction: Clostridium difficile is the major etiological agent of nosocomial antibiotics associated diarrhoea. C. difficile infection is associated with considerable morbidity and mortality among hospitalized patients worldwide. Despite its known importance, there is no study on this important pathogen in Algeria. Methodology: In this prospective study, undertaken between 2013 and 2015, faecal specimens were collected from 159 hospitalized patients with antibiotic-associated diarrhoea in two tertiary health care hospitals in Chlef, Algeria. Faecal samples were cultured on CLO plates Agar with cefoxitin, cycloserine antibiotics and sodium taurocholate. C. difficile suspected colonies were analysed by multiplex PCR for the detection of the toxin genes. C. difficile isolates were analysed by PCR ribotyping and multi-locus tandem repeat analysis. Antimicrobial susceptibility was determined by the E-test method, according to the Clinical and Laboratory Standards Institute protocol. Results: C. difficile was cultured from 11 of 159 stool specimen (6.9%). Seven strains were toxigenic, mainly represented by the 020 and 014 PCR ribotypes and four non toxigenic belong to PCR ribotype 084. All 11 isolates were susceptible to both vancomycin and metronidazole and resistant to ciprofloxacin. Conclusions: This study, which reported for the first time C. difficile ribotypes circulating in Algerian health care facilities, could paves the way for further more comprehensive studies on this important pathogen, and could be useful to the local health authorities to implement a surveillance program of C. difficile in Algeria.

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Inactivation of Clostridium difficile spore outgrowth by synergistic effects of nisin and lysozyme

Canadian Journal of Microbiology ◽

10.1139/cjm-2016-0550 ◽

2017 ◽

Vol 63 (7) ◽

pp. 638-643 ◽

Cited By ~ 4

Author(s):

Changhoon Chai ◽

Kyung-Soo Lee ◽

Goo-Sang Imm ◽

Young Soon Kim ◽

Se-Wook Oh

Keyword(s):

Clostridium Difficile ◽

Synergistic Effects ◽

Sodium Taurocholate ◽

Natural Antimicrobials ◽

Human Contact ◽

Spore Outgrowth

Inactivating Clostridium difficile spores is difficult, as they are resistant to heat, chemicals, and antimicrobials. However, this note describes inactivation of C. difficile spore outgrowth by incubation in a solution containing a germinant (1% (m/v) sodium taurocholate), co-germinants (1% (m/v) tryptose and 1% (m/v) NaCl), and natural antimicrobials (20 nmol·L–1 nisin and 0.2 mmol·L–1 lysozyme). Clostridium difficile spores were resistant to nisin and lysozyme but became susceptible during germination and outgrowth triggered and promoted by sodium taurocholate, tryptose, and NaCl. The degree of inactivation of germinated and outgrowing C. difficile spores by both nisin and lysozyme was greater than the sum of that by nisin and lysozyme individually, suggesting synergistic inactivation of C. difficile spores. The germinant, co-germinants, and natural antimicrobials used in this study are safe for human contact and consumption. Therefore, these findings will facilitate the development of a safe and effective method to inactivate C. difficile spore.

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