Toxigenic Clostridium difficile in retail packed chicken meat and broiler flocks in northeastern Iran

Document Type: Full paper (Original article)

Authors

1 Department of Clinical Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran

2 Department of Poultry Diseases, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

3 Department of Food Hygiene and Aquaculture, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran

Abstract

This study was designed to evaluate the occurrence of Clostridium difficile in both broiler chicken farms and packed chicken parts sold at market places in Mashhad, the second most popular Islamic pilgrimage city after Mecca in northeastern Iran. The fresh faecal samples were obtained from broiler farms, while the chicken packs were purchased from retail outlets across the city at market places and samples were obtained from the necks, thighs, and wings. The selective culture was used for isolation of C. difficile. Out of 40 pooled fresh faecal and 65 packed chicken samples, 14 (35%) and 10 (15.3%) samples were positive, respectively. Some of the C. difficile isolates from chickens packs (7 out of 10, 70%) and faecal samples (5 out of 14, 36%) were detected as toxigenic (A, B and binary toxins) using molecular identification. From 14 isolates of the faecal samples, five isolates were tcdA and tcdB positive, and none was binary toxin positive. The results of the present study suggest that broiler chickens are a potential source of C. difficile, which may infect humans through contact or consumption of chicken meat, although the significance of food contamination is entirely unclear, the role of poultry products as a potential source of the infection should be investigated.

Keywords


Antikainen, J; Pasanen, T; Mero, S; Tarkka, E; Kirveskari, J; Kotila, S; Mentula, S; Kononen, E; Virolainen-Julkunen, A; Vaara, M and Tissari, P (2009). Detection of virulence genes of Clostridium difficile by multiplex PCR. APMIS., 117: 607-613.

Bakker, D; Smits, WK; Kuijper, EDJ and Corver, J (2012). TcdC does not significantly repress toxin expression in Clostridium difficile 630 DeltaErm. PLoS One. 7: e43247.

Bartlett, JG (1992). Antibiotic-associated diarrhea. Clin. Infect. Dis., 15: 573-581.

Bidet, P; Barbut, F; Lalande, V; Burghoffer, B and Petit, J (1999). Development of a new PCR-ribotyping method for Clostridium difficile based on ribosomal RNA gene sequencing. FEMS. Microbiol. Lett., 175: 261-266.

Curry, SR; Marsh, JW; Muto, CA; O’leary, MM; Pasculle, AW and Harrison, LH (2007). TcdC genotypes associated with severe TcdC truncation in an epidemic clone and other strains of Clostridium difficile. J. Clin. Microbiol., 45: 215-221.

Curry, SR; Marsh, JW; Schlackman, JL and Harrison, LH (2012). Prevalence of Clostridium difficile in uncooked ground meat products from Pittsburgh, Pennsylvania. Appl. Environ. Microbiol., 78: 4183-4186.

Dupuy, B; Govind, R; Antunes, A and Matamouros, S (2008). Clostridium difficile toxin synthesis is negatively regulated by TcdC. J. Med. Microbiol., 57: 685-689.

Fedorko, DP and Williams, EC (1997). Use of cycloserine-cefoxitin-fructose agar and L-proline-aminopeptidase (PRO Discs) in the rapid identification of Clostridium difficile. J. Clin. Microbiol., 35: 1258-1259.

Ghavidel, M; Salari Sedigh, H and Razmyar, J (2016). Isolation of Clostridium difficile and molecular detection of binary and A/B toxins in faeces of dogs. Iran. J. Vet. Res., 17: 273-276.

Gould, LH and Limbago, B (2010). Clostridium difficile in food and domestic animals: a new foodborne pathogen? Clin. Infect. Dis., 51: 577-582.

Guran, HS and Ilhak, OI (2015). Clostridium difficile in retail chicken meat parts and liver in the Eastern Region of Turkey. J. Consum. Protect. Food Safety. 10: 359-364.

Harvey, RB; Norman, KN; Andrews, K; Hume, ME; Scanlan, CM; Callaway, TR; Anderson, RC and Nisbet, DJ (2011a). Clostridium difficile in poultry and poultry meat. Foodborne Pathog. Dis., 8: 1321-1323.

Harvey, RB; Norman, KN; Andrews, K; Norby, B; Hume, ME; Scanlan, CM; Hardin, MD and Scott, HM (2011b). Clostridium difficile in retail meat and processing plants in Texas. J. Vet. Diagn. Invest., 23: 807-811.

Indra, A; Schmid, D; Huhulescu, S; Hell, M; Gattringer, R; Hasenberger, P; Fiedler, A; Wewalka, G and Allerberger, F (2008). Characterization of clinical Clostridium difficile isolates by PCR ribotyping and detection of toxin genes in Austria, 2006-2007. J. Med. Microbiol., 57: 702-708.

Jung, KS; Park, JJ; Chon, YE; Jung, ES; Lee, HJ; Jang, HW; Lee, KJ; Lee, SH; Moon, CM; Lee, JH; Shin, JK; Jeon, SM; Hong, SP; Kim, TI; Kim, WH and Cheon, JH (2010). Risk factors for treatment failure and recurrence after metronidazole treatment for Clostridium difficile-associated diarrhea. Gut liver. 4: 332-337.

Kato, H; Kato, N; Watanabe, K; Iwai, N; Nakamura, H; Yamamoto, T; Suzuki, K; Kim, S; Chong, Y and Wasito, EB (1998). Identification of toxin A-negative, toxin B-positive Clostridium difficile by PCR. J. Clin. Microbial., 36: 2178-2182.

Lemee, L; Dhalluin, A; Pestel-Caron, M; Lemeland, JF and Pons, JL (2004). Multilocus sequence typing analysis of human and animal Clostridium difficile isolates of various toxigenic types. J. Clin. Microbiol., 42: 2609-2617.

Matamouros, S; England, P and Dupuy, B (2007). Clostridium difficile toxin expression is inhibited by the novel regulator TcdC. Mol. Microbiol., 64: 1274-1288.

Merrigan, M; Venugopal, A; Mallozzi, M; Roxas, B; Viswanathan, VK; Johnson, S; Gerding, DN and Vedantam, G (2010). Human hypervirulent Clostridium difficile strains exhibit increased sporulation as well as robust toxin production. J. Bacteriol., 192: 4904-4911.

Persson, S; Torpdahl, M and Olsen, KE (2008). New multiplex PCR method for the detection of Clostridium difficile toxin A (tcdA) and toxin B (tcdB) and the binary toxin (cdtA⁄cdtB) genes applied to a Danish strain collection. Clin. Microbiol. Infect., 14: 1057-1064.

Pituch, H; Obuch-Woszczatyñski, P; Van Den Braak, N;
Van Belkum, A; Kujawa, M; Luczak, M and Meisel-
Mikolajczy, F (2002). Variable flagella expression among clonal toxin A/B+ Clostridium difficile strains with highly homogeneous flagellin genes. Clin. Microbiol. Infect., 8: 187-188.

Popoff, MR; Rubin, EJ; Gill, DM and Boquet, P (1988). Actin-specific ADP-ribosyltransferase produced by a Clostridium difficile strain. Infect. Immun., 56: 2299-2306.

Rodriguez-Palacios, A; Staempfli, HR; Duffield, T and Weese, JS (2007). Clostridium difficile in retail ground meat, Canada. Emerg. Infect. Dis., 13: 485-487.

Rupnik, M (2007). Is Clostridium difficile-associated infection a potentially zoonotic and foodborne disease? Clin. Microbiol. Infect., 13: 457-459.

Simango, C (2006). Prevalence of Clostridium difficile in the environment in a rural community in Zimbabwe. T. Roy. Soc. Trop. Med. H., 100: 1146-1150.

Simango, C and Mwakurudza, S (2008). Clostridium difficile in broiler chickens sold at market places in Zimbabwe and their antimicrobial susceptibility. Int. J. Food Microbiol., 124:268-270.

Songer, JG and Anderson, MA (2006). Clostridium difficile: an important pathogen of food animals. Anaerobe. 12: 1-4.

Songer, JG; Trinh, HT; Killgore, GE; Thompson, AD; Mcdonald, LC and Limbago, BM (2009). Clostridium difficile in retail meat products, USA, 2007. Emerg. Infect. Dis., 15: 819-821.

Stubbs, SL; Brazier, JS; O’neill, GL and Duerden, BH (1999). PCR targeted to the 16S-23S rRNA gene intergenic spacer region of Clostridium difficile and construction of a library consisting of 116 different PCR ribotypes. J. Clin. Microbiol., 37: 461-463.

Viscidi, R; Willey, S and Bartlett, JG (1981). Isolation rates and toxigenic potential of Clostridium difficile isolates from various patient populations. Gastroenterology. 81: 5-9.

Vohra, P and Poxton, IR (2011). Comparison of toxin and spore production in clinically relevant strains of Clostridium difficile. Microbiology. 157: 1343-1353.

Weese, JS; Avery, BP; Rousseau, J and Reid-Smith, RJ (2009). Detection and enumeration of Clostridium difficile spores in retail beef and pork. Appl. Environ. Microbiol., 75: 5009-5011.

Weese, JS; Reid-Smith, RJ; Avery, BP and Rousseau, J (2010). Detection and characterization of Clostridium difficile in retail chicken. Lett. Appl. Microbiol., 50: 362-365.