Short communicationDevelopment of a consensus method for culture of Clostridium difficile from meat and its use in a survey of U.S. retail meats
Highlights
► A standard protocol for Clostridium difficile culture from retail meats was established. ► 1755 retail meats were cultured for C. difficile over a 12 month period. ► No C. difficile was isolated from meat, although other Clostridium ssp. were common.
Introduction
Clostridium difficile is an important cause of infectious diarrhea in healthcare settings, usually following antimicrobial therapy. However, C. difficile infection (CDI) is an increasingly-recognized cause of diarrhea among people in community settings without recent inpatient hospital exposure (Centers for Disease Control and Prevention, 2008; Kutty et al., 2010; Lambert et al., 2009). Hypervirulent C. difficile strains have been associated with increased incidence and severity of CDI in healthcare settings (Deneve et al., 2009; McDonald et al., 2005); increases in community-associated CDI (CA-CDI) may be driven by other factors. C. difficile causes disease in food animals (Debast et al., 2009; Keel et al., 2007; Songer, 2004) and has been recovered from retail foods in several countries (de Boer et al., 2011; Harvey et al., 2011a, 2011b; Johnson et al., 2005; Metcalf et al., 2010; Rodriguez-Palacios et al., 2007; Songer et al., 2009), leading some to suggest that food may be a source for CA-CDI (Metcalf et al., 2010; Rupnik, 2007; Rupnik and Songer, 2010). Although several groups have reported the isolation of C. difficile from retail meats (Harvey et al., 2011a, 2011b; Rodriguez-Palacios et al., 2009; Rodriguez-Palacios et al., 2007; Songer et al., 2009; Weese et al., 2009), there is no consensus method for culture of C. difficile from meats or other food products. This study was designed to establish a consensus method for culture of C. difficile from meats, and to determine the prevalence of C. difficile contamination of selected retail meats in the United States using a standardized culture method.
Section snippets
Comparison of culture methods for C. difficile culture from meats
Several different culture methods were evaluated for recovery of C. difficile from spiked ground meat samples. Independently-acquired ground beef was inoculated using a single spore suspension at a rate of approximately 100 spores/gram in three laboratories with expertise culturing and characterizing C. difficile. The spore suspension was prepared as described previously (Bertolo et al., 2012), except that phase contrast microscopy was not performed. Three types of initial broth enrichment were
Selection of a consensus method for culture of C. difficile from meats
C. difficile was recovered from the spiked meat samples with each of the methods evaluated. The best recovery was observed after 3 and 5 days of enrichment in broth medium (Table 1). Recovery of C. difficile in BHIT broth without initial heat shock appeared to be better than in CDMN broth or in BHIT with initial heat shock (Table 1). A subsequent evaluation of BHI compared with BHIT demonstrated no added benefit of taurocholate on C. difficile recovery in the broth enrichment step (data not
Disclaimer
The findings and conclusions in this report are those of the author(s) and do not necessarily represent the official position of the Centers for Disease Control and Prevention.
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2019, Veterinary MicrobiologyCitation Excerpt :Similar to what has been shown for Salmonella spp., (Palmer et al., 1985) C. difficile and C. perfringens cannot be isolated from feces consistently, as shedding is intermittent or numbers are too low for cultural detection. Detection threshold for C. perfringens has been shown to be 9 cfu/g feces using this method ;(Schoster et al., 2012) several different methods have been shown to be able to detect 100 spores/g feces of C. difficile, including the method used here (Limbago et al., 2012). We could show that multiple sampling increased the prevalence of Clostridia spp. from 1 to 10% on a single given day, up to 19% when samples were collected over three days.
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2019, Food MicrobiologyCitation Excerpt :Selective antibiotics employed in solid medium, such cefoxitin and cycloserine (CCFA; Indra et al., 2009) or moxalactam and norfloxacin (CDMN; Weese et al., 2009), could also be affecting C. difficile isolation. A consensus protocol using 10 g of sample in 90 mL of BHI enrichment broth supplemented with taurocholate, for 3–5 days, followed by ethanol shock and plating to selective and non-selective media was proposed for the isolation of C. difficile (Limbago et al., 2012). Traditional culture provides an isolate for further typing methods, wich are essential for epidemiological studies (Avbersek, 2017; Knetsch et al., 2013).
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2017, Advances in Food and Nutrition ResearchClostridium difficile infection: Early history, diagnosis and molecular strain typing methods
2016, Microbial PathogenesisCitation Excerpt :Furthermore, they are used for the clinical recovery of C. difficile from faecal samples and not for the semi-quantification of viable spores [160]. Pre-treatment of samples with ethanol shock has been associated with an increase in sensitivity [161–164]. However, in the different studies conducted in our laboratory (unpublished data), ethanol shock or pre-heat treatment of samples does not improve the recovery of C. difficile from faecal or food samples.
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Current affiliation: The Carter Center, Atlanta, GA, USA.