title: Prevalence and molecular characterization of Clostridium difficile isolates from a pig slaughterhouse, pork, and humans in Taiwan.
abstract: Clostridium difficile causes antibiotic-associated diarrhea in both humans and animals. The ribotype 078, predominant in food animals, is associated with community-acquired C. difficile infection, and C. difficile is suggested to be a foodborne pathogen. Recently, the C. difficile ribotype 078 lineage emerged in patients and pigs in Taiwan. This study aimed to investigate the prevalence and molecular characterization of C. difficile isolated from a pig slaughterhouse, retail meat, ready-to-eat meals, and humans in Taiwan. We collected samples from one slaughterhouse (n=422), 29 retail markets (raw pork, n=62; ready-to-eat pork, n=65), and one hospital (non-diarrheal humans, stool, n=317) in 2015. The isolated C. difficile were subjected to ribotyping and multilocus variable-number tandem-repeat analysis (MLVA). In the slaughterhouse, the isolation rate from carcasses was high (23%, 21/92) and ribotype 126 dominated. Scalding water was found to have C. difficile contamination (44%, 4/9), and two of the seven isolates were ribotype 126. The isolation rates from raw pork and ready-to-eat pork were between 20% and 29%. Ribotypes 126, 127, and 014 were found in raw pork, whereas ribotype 078 was not identified in this study. Eight isolates-seven non-toxigenic isolates and one ribotype 017-were found in non-diarrheal human samples. Notably, MLVA showed that ribotype 126 isolates from the slaughterhouse, pig stool, colons, carcasses, and scalding water were closely genetically related, indicating serious risk for cross-contamination. However, the genetic evidence of foodborne transmission from carcasses to food and humans is still lacking.
authors.affiliation: Graduate Institute of Veterinary Pathobiology, College of Veterinary Medicine, National Chung Hsing University, No. 250, Guoguang Rd., South Dist., Taichung 402, Taiwan. Electronic address: pata0112@gmail.com.
authors.forename: Ying-Chen
authors.initials: YC
authors.lastname: Wu
authors.affiliation: Division of Infectious Disease, Department of Internal Medicine, Tungs' Taichung MetroHarbor Hospital, No.699, Sec. 8, Taiwan Blvd., Taichung 435, Taiwan; Department of Health Food, Chung Chou University of Science and Technology, No. 6, Lane 2, Sec. 3, Shanjiao Rd., Yuanlin Township, Changhua 510, Taiwan. Electronic address: chioming2002@yahoo.com.tw.
authors.forename: Chih-Ming
authors.initials: CM
authors.lastname: Chen
authors.affiliation: Department of Veterinary Medicine, National Chung Hsing University, No. 250, Guoguang Rd., South Dist., Taichung 402, Taiwan. Electronic address: ck476@nchu.edu.tw.
authors.forename: Chih-Jung
authors.initials: CJ
authors.lastname: Kuo
authors.affiliation: Graduate Institute of Veterinary Pathobiology, College of Veterinary Medicine, National Chung Hsing University, No. 250, Guoguang Rd., South Dist., Taichung 402, Taiwan. Electronic address: frinlee@hotmail.com.
authors.forename: Jen-Jie
authors.initials: JJ
authors.lastname: Lee
authors.affiliation: Graduate Institute of Microbiology and Public Health, College of Veterinary Medicine, National Chung Hsing University, No. 250, Guoguang Rd., South Dist., Taichung 402, Taiwan. Electronic address: 220aall@gmail.com.
authors.forename: Pin-Chun
authors.initials: PC
authors.lastname: Chen
authors.affiliation: Department of Medical Laboratory Science and Biotechnology, China Medical University, No. 91 Hsuehshih Rd., Taichung 402, Taiwan. Electronic address: yichih@mail.cmu.edu.tw.
authors.forename: Yi-Chih
authors.initials: YC
authors.lastname: Chang
authors.affiliation: Graduate Institute of Veterinary Pathobiology, College of Veterinary Medicine, National Chung Hsing University, No. 250, Guoguang Rd., South Dist., Taichung 402, Taiwan. Electronic address: thc@dragon.nchu.edu.tw.
authors.forename: Ter-Hsin
authors.initials: TH
authors.lastname: Chen
mesh_terms: D000003:Abattoirs / Q000706:statistics & numerical data; D000818:Animals; D016360:Clostridioides difficile / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D003015:Clostridium Infections / Q000382:microbiology; D057140:Fast Foods / Q000191:economics / Q000382:microbiology; D005243:Feces / Q000382:microbiology; D005260:Female; D005506:Food Contamination / Q000032:analysis*; D005511:Food Handling; D006801:Humans; D015995:Prevalence; D000069466:Red Meat / Q000032:analysis / Q000382:microbiology*; D021521:Ribotyping; D013552:Swine; D013624:Taiwan
keywords: Cefoxitin (PubChem CID: 441199); Clostridium difficile; Hemin (PubChem CID: 455658); Lysozyme (PubChem CID: 16129749); Metronidazole (PubChem CID: 4173); Moxifloxacin (PubChem CID: 152946); Pig; Pork; Ribotype 126; Ribotype 127; Slaughterhouse; Sodium bicarbonate (PubChem CID: 516892); Sodium taurocholate (PubChem CID: 23666345); Thioglycolic acid (PubChem CID: 1133); Vancomycin (PubChem CID: 14969); Vitamin K(1) (PubChem CID: 5284607); d-cycloserine (PubChem CID: 6234); d-mannitol (PubChem CID: 6251); l-cysteine (PubChem CID:5862)