Comparison of alcohol shock enrichment and selective enrichment for the isolation of<i>Clostridium difficile</i>

Riley, Thomas V.; Brazier, Jon S.; Hassan, Hamimah; Williams, Kathleen; Phillips, K. D.

doi:10.1017/s0950268800067832

Cited by 23 publications

(13 citation statements)

References 17 publications

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“…Total C. difficile cells (vegetative cells and spores) or only spores (ethanol resistant) in fecal samples can easily be enumerated using CCEY selective medium in combination with the ethanol shock method (47). We found that immediately after excretion 10 to 20% of the total C. difficile vegetative cells and spores shed in the feces of supershedders were spores (Fig.…”

Section: Resultsmentioning

confidence: 84%

Antibiotic Treatment of Clostridium difficile Carrier Mice Triggers a Supershedder State, Spore-Mediated Transmission, and Severe Disease in Immunocompromised Hosts

et al. 2009

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Clostridium difficile persists in hospitals by exploiting an infection cycle that is dependent on humans shedding highly resistant and infectious spores. Here we show that human virulent C. difficile can asymptomatically colonize the intestines of immunocompetent mice, establishing a carrier state that persists for many months. C. difficile carrier mice consistently shed low levels of spores but, surprisingly, do not transmit infection to cohabiting mice. However, antibiotic treatment of carriers triggers a highly contagious supershedder state, characterized by a dramatic reduction in the intestinal microbiota species diversity, C. difficile overgrowth, and excretion of high levels of spores. Stopping antibiotic treatment normally leads to recovery of the intestinal microbiota species diversity and suppresses C. difficile levels, although some mice persist in the supershedding state for extended periods. Spore-mediated transmission to immunocompetent mice treated with antibiotics results in self-limiting mucosal inflammation of the large intestine. In contrast, transmission to mice whose innate immune responses are compromised (Myd88 ؊/؊ ) leads to a severe intestinal disease that is often fatal. Thus, mice can be used to investigate distinct stages of the C. difficile infection cycle and can serve as a valuable surrogate for studying the spore-mediated transmission and interactions between C. difficile and the host and its microbiota, and the results obtained should guide infection control measures.

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Section: Resultsmentioning

confidence: 84%

Antibiotic Treatment of Clostridium difficile Carrier Mice Triggers a Supershedder State, Spore-Mediated Transmission, and Severe Disease in Immunocompromised Hosts

et al. 2009

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“…Enrichment culture, in addition to culture on solid media, was used in the present study, while the earlier work relied on the use of solid media (CCFA with 0.1 % sodium taurocholate) and an alcohol shock procedure [3]. In a previous study [14] comparing enrichment culture and the alcohol shock procedure, enrichment was significantly better than alcohol shock.…”

Section: Resultsmentioning

confidence: 99%

Gastrointestinal carriage ofClostridium difficilein cats and dogs attending veterinary clinics

et al. 1991

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SUMMARYCats and dogs being treated at two veterinary clinics were investigated for gastrointestinal carriage of Clostridium difficile using selective solid and enrichment media. Thirty-two (39 5 %) of 81 stool samples yielded C. difficile. There were significant differences in isolation rates between clinics, 610 % of animals being positive at one clinic compared to 17-5 % at the other (Chi-square, P < 0005). Of 29 animals receiving antibiotics, 15 (52-0 %) harboured C. difficile while 11 (23 9 %) of 46 animals not receiving antibiotics were positive (Chi-square, P < 00 1). There was no difference in carriage rate between cats (38 1 %) and dogs (40 0 %). The environment at both veterinary clinics was surveyed for the presence of C. difficile. Fifteen of 20 sites at one clinic were positive compared to 6 of 14 sites at the other clinic. Both cytotoxigenic and noncytotoxigenic isolates of C. difficile were recovered from animals and environmental sites. These findings suggest that household pets may be a potentially significant reservoir of infection with C. difficile.

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“…To date, there is no consensus on the best method for recovery of the organism in culture. Some investigators have employed a heat shock or alcohol shock prior to inoculation to enhance spores of C. difficile and inhibit growth of other organisms (66)(67)(68). A variety of different medium formulations have been described, many of which take advantage of the fact that C. difficile can ferment fructose (69,70).…”

Section: Laboratory Methods For Clostridium Difficile Detection Methodsmentioning

confidence: 99%

Diagnosis of Clostridium difficile Infection: an Ongoing Conundrum for Clinicians and for Clinical Laboratories

2013

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SUMMARY Clostridium difficile is a formidable nosocomial and community-acquired pathogen, causing clinical presentations ranging from asymptomatic colonization to self-limiting diarrhea to toxic megacolon and fulminant colitis. Since the early 2000s, the incidence of C. difficile disease has increased dramatically, and this is thought to be due to the emergence of new strain types. For many years, the mainstay of C. difficile disease diagnosis was enzyme immunoassays for detection of the C. difficile toxin(s), although it is now generally accepted that these assays lack sensitivity. A number of molecular assays are commercially available for the detection of C. difficile . This review covers the history and biology of C. difficile and provides an in-depth discussion of the laboratory methods used for the diagnosis of C. difficile infection (CDI). In addition, strain typing methods for C. difficile and the evolving epidemiology of colonization and infection with this organism are discussed. Finally, considerations for diagnosing C. difficile disease in special patient populations, such as children, oncology patients, transplant patients, and patients with inflammatory bowel disease, are described. As detection of C. difficile in clinical specimens does not always equate with disease, the diagnosis of C. difficile infection continues to be a challenge for both laboratories and clinicians.

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Comparison of alcohol shock enrichment and selective enrichment for the isolation ofClostridium difficile

Cited by 23 publications

References 17 publications

Antibiotic Treatment of Clostridium difficile Carrier Mice Triggers a Supershedder State, Spore-Mediated Transmission, and Severe Disease in Immunocompromised Hosts

Antibiotic Treatment of Clostridium difficile Carrier Mice Triggers a Supershedder State, Spore-Mediated Transmission, and Severe Disease in Immunocompromised Hosts

Gastrointestinal carriage ofClostridium difficilein cats and dogs attending veterinary clinics

Diagnosis of Clostridium difficile Infection: an Ongoing Conundrum for Clinicians and for Clinical Laboratories

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