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Characterization of a multidrug resistant C. difficile meat isolate.

International journal of food microbiology (2014-12-03)
Shankumar Mooyottu, Genevieve Flock, Anup Kollanoor-Johny, Indu Upadhyaya, Bhushan Jayarao, Kumar Venkitanarayanan
ABSTRACT

Clostridium difficile is a pathogen of significant public health concern causing a life-threatening, toxin-mediated enteric disease in humans. The incidence and severity of the disease associated with C. difficile have increased in the US with the emergence of hypervirulent strains and community associated outbreaks. The detection of genotypically similar and identical C. difficile strains implicated from human infections in foods and food animals indicates the potential role of food as a source of community associated C. difficile disease. One hundred samples each of ground beef, pork and chicken obtained from geographically distant grocery stores in Connecticut were tested for C. difficile. Positive isolates were characterized by ribotyping, antibiotic susceptibility, toxin production and whole genome sequencing. Of the 300 meat samples, only two pork samples tested positive for C. difficile indicating a very low prevalence of C. difficile in meat. The isolates were non toxigenic; however, genome characterization revealed the presence of several antibiotic resistance genes and mobile elements that can potentially contribute to generation of multidrug resistant toxigenic C. difficile by horizontal gene transfer. Further studies are warranted to investigate potential food-borne transmission of the meat isolates and development of multi-drug resistance in these strains.

MATERIALS
Product Number
Brand
Product Description

USP
Ampicillin, United States Pharmacopeia (USP) Reference Standard
Supelco
Ciprofloxacin, Pharmaceutical Secondary Standard; Certified Reference Material
Ampicillin, anhydrous, European Pharmacopoeia (EP) Reference Standard
Norfloxacin, European Pharmacopoeia (EP) Reference Standard
Ciprofloxacin, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Ampicillin, anhydrous, 96.0-102.0% (anhydrous basis)
Supelco
Norfloxacin, analytical standard, ≥98% (TLC)
Supelco
Norfloxacin, VETRANAL®, analytical standard
Supelco
Ciprofloxacin, VETRANAL®, analytical standard
Sigma-Aldrich
Ciprofloxacin, ≥98% (HPLC)
Rifampicin, European Pharmacopoeia (EP) Reference Standard
Supelco
Ampicillin
Norfloxacin for peak identification, European Pharmacopoeia (EP) Reference Standard
USP
L-Proline, United States Pharmacopeia (USP) Reference Standard
Supelco
Metronidazole, Pharmaceutical Secondary Standard; Certified Reference Material
USP
Erythromycin, United States Pharmacopeia (USP) Reference Standard
Erythromycin, for microbiological assay, European Pharmacopoeia (EP) Reference Standard
Metronidazole, European Pharmacopoeia (EP) Reference Standard
Supelco
Erythromycin, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Erythromycin standard solution, 1 mg/mL in H2O
Sigma-Aldrich
Ampicillin, meets USP testing specifications
Sigma-Aldrich
Rifampicin, ≥95% (HPLC), powder or crystals
SAFC
L-Proline
Sodium taurocholate, BRP, European Pharmacopoeia (EP) Reference Standard
Supelco
L-Proline, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Rifampicin, suitable for plant cell culture, BioReagent, ≥95% (HPLC), powder or crystals
Supelco
L-Proline, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Sigma-Aldrich
L-Proline, BioUltra, ≥99.5% (NT)
Sigma-Aldrich
L-Proline, from non-animal source, meets EP, USP testing specifications, suitable for cell culture
Sigma-Aldrich
L-Proline, ReagentPlus®, ≥99% (HPLC)