Characterization of a multidrug resistant C. difficile meat isolate.

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.

[1]  Jane W. Marsh,et al.  Multi-locus variable number tandem repeat analysis for investigation of the genetic association of Clostridium difficile isolates from food, food animals and humans. , 2011, Anaerobe.

[2]  E. Kuijper,et al.  Clostridium difficile PCR ribotype 078 toxinotype V found in diarrhoeal pigs identical to isolates from affected humans. , 2009, Environmental microbiology.

[3]  C. E. MacGowan,et al.  Development of a consensus method for culture of Clostridium difficile from meat and its use in a survey of U.S. retail meats. , 2012, Food microbiology.

[4]  B. Dupuy,et al.  Clostridium difficile toxin synthesis is negatively regulated by TcdC. , 2008, Journal of medical microbiology.

[5]  J. Weese,et al.  Detection and Enumeration of Clostridium difficile Spores in Retail Beef and Pork , 2009, Applied and Environmental Microbiology.

[6]  J. Frank,et al.  Clostridium difficile from healthy food animals: optimized isolation and prevalence. , 2011, Journal of food protection.

[7]  P. Mastrantonio,et al.  ErmB Determinants and Tn916-Like Elements in Clinical Isolates of Clostridium difficile , 2005, Antimicrobial Agents and Chemotherapy.

[8]  R. Harvey,et al.  Clostridium difficile in poultry and poultry meat. , 2011, Foodborne pathogens and disease.

[9]  X. Didelot,et al.  Evolutionary History of the Clostridium difficile Pathogenicity Locus , 2013, Genome biology and evolution.

[10]  Mark H. Wilcox,et al.  Clostridium difficile infection: new developments in epidemiology and pathogenesis , 2009, Nature Reviews Microbiology.

[11]  S Kobayashi,et al.  Spread of a large plasmid carrying the cpe gene and the tcp locus amongst Clostridium perfringens isolates from nosocomial outbreaks and sporadic cases of gastroenteritis in a geriatric hospital , 2008, Epidemiology and Infection.

[12]  T. Pasanen,et al.  Detection of virulence genes of Clostridium difficile by multiplex PCR , 2009, APMIS : acta pathologica, microbiologica, et immunologica Scandinavica.

[13]  B. Limbago,et al.  Clostridium difficile in Retail Meat Products, USA, 2007 , 2009, Emerging infectious diseases.

[14]  J. Lejeune,et al.  Clostridium difficile in foods and animals: history and measures to reduce exposure , 2013, Animal Health Research Reviews.

[15]  M. Rupnik,et al.  Clostridium difficile genotypes other than ribotype 078 that are prevalent among human, animal and environmental isolates , 2012, BMC Microbiology.

[16]  Julian Parkhill,et al.  The multidrug-resistant human pathogen Clostridium difficile has a highly mobile, mosaic genome , 2006, Nature Genetics.

[17]  N. Woodford,et al.  Editorial: assessing the antimicrobial susceptibility of bacteria obtained from animals. , 2010, The Journal of antimicrobial chemotherapy.

[18]  M. Rupnik,et al.  Antimicrobial susceptibility of animal and human isolates of Clostridium difficile by broth microdilution. , 2013, Journal of medical microbiology.

[19]  Lihua Zhang,et al.  Genetic analysis of Tn916-like elements conferring tetracycline resistance in clinical isolates of Clostridium difficile. , 2014, International journal of antimicrobial agents.

[20]  J. Blanco,et al.  Characterization of swine isolates of Clostridium difficile in Spain: a potential source of epidemic multidrug resistant strains? , 2013, Anaerobe.

[21]  M. Owings,et al.  Clostridium difficile Infection in Patients Discharged from US Short-stay Hospitals, 1996–2003 , 2006, Emerging Infectious Diseases.

[22]  D. Aronoff,et al.  A clinical and epidemiological review of non-toxigenic Clostridium difficile. , 2013, Anaerobe.

[23]  J. Rousseau,et al.  Detection and characterization of Clostridium difficile in retail chicken , 2010, Letters in applied microbiology.

[24]  M. Wagner,et al.  Clostridium difficile in raw products of animal origin. , 2010, International journal of food microbiology.

[25]  J. Barkin,et al.  Clostridium difficile associated infection, diarrhea and colitis. , 2009, World journal of gastroenterology.

[26]  M. Popoff,et al.  Genetic characteristics of toxigenic Clostridia and toxin gene evolution. , 2013, Toxicon : official journal of the International Society on Toxinology.

[27]  J. Bartlett Clostridium difficile infection: pathophysiology and diagnosis. , 1997, Seminars in gastrointestinal disease.

[28]  A. Sonenshein,et al.  Carvacrol and trans-Cinnamaldehyde Reduce Clostridium difficile Toxin Production and Cytotoxicity in Vitro , 2014, International journal of molecular sciences.

[29]  J. Lemeland,et al.  Multiplex PCR Targeting tpi (Triose Phosphate Isomerase), tcdA (Toxin A), and tcdB (Toxin B) Genes for Toxigenic Culture of Clostridium difficile , 2004, Journal of Clinical Microbiology.

[30]  J. Lejeune,et al.  Prevalence, enumeration, and antimicrobial agent resistance of Clostridium difficile in cattle at harvest in the United States. , 2011, Journal of food protection.

[31]  Peter Mullany,et al.  Horizontal gene transfer converts non-toxigenic Clostridium difficile strains into toxin producers , 2013, Nature Communications.

[32]  P. Bidet,et al.  Development of a new PCR-ribotyping method for Clostridium difficile based on ribosomal RNA gene sequencing. , 1999, FEMS microbiology letters.

[33]  Stuart Johnson,et al.  An epidemic, toxin gene-variant strain of Clostridium difficile. , 2005, The New England journal of medicine.

[34]  A. Roberts,et al.  Draft Genome Sequence of the Nontoxigenic Clostridium difficile Strain CD37 , 2012, Journal of bacteriology.

[35]  J. Weese,et al.  Clostridium difficile and methicillin-resistant Staphylococcus aureus shedding by slaughter-age pigs , 2011, BMC veterinary research.

[36]  E. Kuijper,et al.  The relation between farm specific factors and prevalence of Clostridium difficile in slaughter pigs. , 2011, Veterinary microbiology.