Molecular study of Clostridium difficile isolated from beef in Isfahan, Iran

Authors

1 Infectious Diseases and Tropical Medicine Research Center, Isfahan University of Medical Sciences, Isfahan, Iran & Department of Research and Development, Department of Food and Drug, Isfahan University of Medical Sciences, Isfahan, Iran

2 Food Security Research Centre, School of Nutrition and Food Sciences, Isfahan University of Medical Sciences, Isfahan, Iran

3 Department of Food Science and Technology, College of Food Science and Technology, Tehran Science and Research Branch, Islamic Azad University, Tehran, Iran

4 Department of Pathobiology and Centre for Public Health and Zoonses, Ontario Veterinary College, University of Guelph, Guelph, Ontarion, Canada

5 Department of Animal Science, Faculty of Agriculture and Natural Resources, Tehran Science and Research Branch, Islamic Azad University, Tehran, Iran

6 Infectious Diseases and Tropical Medicine Research Center, Isfahan University of Medical Sciences, Isfahan, Iran

Abstract

BACKGROUND: Clostridium difficile (C. difficile) infection is one of the most important diseases in healthcare facilities and community. Ribotypes 027 and 078 are known as hyper-virulent strain of C. difficile in molecular study. PCR-ribotyping is a suitable method to interpret the relation of C. difficile isolated from food and hospital. Objectives: In the present study, the clostridim difficile binary toxin (cdtB) and ribotype pattern evaluated in toxigenic C. difficle isolated from beef. Methods: Detection of cdtB in 12 toxigenic C. difficile (encoding tcdA and tcdB gene) isolated from 100 beef samples was determined through PCR. Afterwards, PCR-ribotyping was performed to examine the ribotype patterns of C. difficile. Results: cdtB gene was not detected in any positive isolate. Ten different patterns were observed in 12 toxigenic isolates. No similarity existed in the ribotypes of our study with ribotypes 027 and 078. Conclusions: Albeit  ribotyp 027 and 078 were not found in our study, the isolation of toxigenic C. difficile with new ribotypes in Iran may indicate the probable hazard of this bacterium in public health. Comprehensive research about C. difficile in different food sources is recommended on a national level.

Keywords


 
Bidet, P., Lalande, V., Salauze, B., Burghoffer, B., Avesani, V., Delmée, M., Rossier, A., Barbut, F., Petit, J.D. (2000) Comparison of PCR-ribotyping, arbitrarily primed PCR, and pulsed-field gel electrophoresis for typing Clostridium difficile. J Clin Microbiol. 38: 2484-2487.
Bouttier, S., Barc, M.C., Felix, B., Lambert, S., Collignon, A., Barbut, F. (2010) Clostridium difficile in ground meat, France. Emerg Infect Dis.16:733-735.
de Boer, E., Zwartkruis-Nahuis, A., Heuvelink, A.E., Harmanus, C., Kuijper, E.J. (2011) Prevalence of Clostridium difficile in retailed meat in the Netherlands. Int J Food Microbiol. 144:561-564.
Esfandiari, Z., Jalali, M., Ezzatpanah, H., Weese, J.S., Chamani, M. (2014) Examination of Clostridium difficile contamination in beef meat distributed in Isfahan using culture and multiplex-PCR method. Biol J Microrganism. 11: 109-116.
Esfandiari, Z., Weese, J.S., Ezzatpanah, H., Jalali, M., Chamani, M. (2014) Occurrence of Clostridium difficile in seasoned hamburgers and seven processing plants in Iran. BMC Microbiol. 14: 283.
Esfandiari, Z., Jalali, M., Ezzatpanah, H., Weese, J.S., Chamani, M. (2014) Prevalence and Characterization of Clostridium difficile in beef and mutton meats of Isfahan region, Iran. Jundishapur J Microbiol. 7: e16771. DOI: 10.5812/jjm.16771.
Esfandiari, Z., Weese, J.S., Ezzatpanah, H., Chamani, M., Shoaei, P., Yaran, M., Ataei, B., Maracy, M.R., Ansariyan, A., Ebrahimi, F., Jalali, M. (2015) Isolation and characterization of Clostridium difficile in farm animals from slaughterhouse to retail stage in Isfahan, Iran. Foodborne Pathog Dis. 12: 10.
Esfandiari, Z., Jalali, M., Safaeian, L., Weese, J.S. A review on epidemiology of Clostridium difficile infection. Tehran Univ Med J. 74: 305-313.
Hasanzade, H., Rahimi, E. (2013) Isolation of Clostridium difficile from turkey and ostrich meat sold in meat stores of Isfahan city. Int J Advanced Biolog Biomed Res. 1: 963-967.
Jalali,  M., Khorvash,  F., Warriner, K., Weese, J.S. (2012)  Clostridium difficile infection in an Iranian hospital. BMC Res Notes. 5: 1-5.
Jank, T., AktoriesK, K. (2008) Structure and mode of action of clostridial glucosylating toxins: the ABCD model. Trends Microbiol. 16:222-229.
Jhung, M.A., Thompson, A.D., Killgore, G.E., Zukowski, W.E., Songer, G., Warny,  M., Johnson, S., Gerding, D.N., McDonald, L.C., Limbago, B.M.  (2008) Toxinotype V Clostridium difficile in humans and food animals. Emerg  Infect Dis.14:1039-1045.
Jobstl, M., Heuberger, S., Indra, A., Nepf, R., Kofer, J., Wagner, M. (2010) Clos‌tridium difficile in raw products of animal origin. Int J Food Microbiol. 138:172-175.
Keel, K., Brazier, J.S., Post, K.W.,  Weese, S., Songer, J.G. (2007) Prevalence of PCR ribotypes among Clostridium difficile isolates from pigs, calves and other species. J Clin Microbiol. 45. 1963-1964.
Kuehne, S.A., Cartman, S.T. (2011) Both, toxin A and toxin B are important in Clostridium difficile infection. Gut Microbes. 2: 252-255.
Kutty, P.K., Woods, C.W., Sena, A.C., Benoit, S.R., Naggie, S., Frederick, J., Evans, S., Engel, J., McDonald, L.C.  (2010)  Risk factors for and estimated incidence of community-associated Clostridium difficile infection, North Carolina, USA. Emerg Infect  Dis.16:197-204.
Popoff, M.R., Rubin, E.J., Gill, D.M., Boquet, P. (1988)  Actin-specific ADP-ribosyltransferase produced by a Clostridium difficile strain. Infect Immun. 56, 2299-2306.
Quesada-Gomez, C., Mulvey, M.R., Vargas, P., Gamboa-Coronado Mdel, M., Rodriguez, C., Rodriguez-Cavillini, E. (2013) Isolation of a toxigenic and clinical genotype of clostridium difficile in retail meats in Costa Rica. J Food Prot. 76:348-351.
Rahimi, E., Jalali, M., Weese, J.S. (2014) Prevalence of Clostridium difficile in raw beef, cow, sheep, goat, camel and buffalo meat in Iran. BMC Public Health. 14:119. 10.1186/1471-2458-14-119.
Rahimi, E., Momtaz, H., Hemmati, M. (2014) Occurrence of Clostridium difficile in raw bovine, ovine, caprine, camel and buffalo milk in Iran. Kafkas Univ Vet Fak Derg. 20: 371-374.
Rahimi, E., Khaksar, F. (2015) Detection of toxigenic Clostidium difficile strains isolated from meat and meat products in Iran. Bulgarian J Vet Med. 18:277-281.
Rahimi, E., Sadat Afzali, Z., Torki Baghbadorani, Z. (2015) Clostridium diffiicle in ready-to-eat foods in Isfahan and Shahrekord, Iran. Asian Pac J Trop Biomed. 5: 128-131.
Rodriguez-Palacios, A., Staempfli, H.R., Duffield, T., Weese, J.S. (2007) Clostridium difficile in retail ground meat, Canada. Emerg Infect Dis. 13:485-487.
Rodriguez-Palacios, A., Reid-Smith, R.J.,  Staempfli, H.R., Daignault, D.,  Janecko, N.,  Avery, B.P., Martin, H., Thomspon, A.D., McDonald, L.C., Limbago, B., Weese, J.S. (2009) Possible seasonality of Clostridium difficile in retail meat, Canada. Emerg Infect Dis. 15: 802-805.
Songer, J.G., Trinh, H.T., Killgore, G.E., Thompson, A.D., McDonald, L.C., Limbago, B.M. (2009) Clostridium difficile in retail meat products, USA, 2007. Emerg Infect Dis. 15:819-21.
Stubbs, S., Rupnik, M., Gibert, M., Brazier, J., Duerden, B., Popoff, M. (2000) Production of actin-specific ADP-ribosyltransferase (binary toxin) by strains of Clostridium difficile. FEMS Microbiol Lett. 186:307 312.
Warny, M., Pepin, J., Fang, A., Killgore, G., Thompson, A., Brazier, J., Frost, E., McDonald, L.C. (2005) Toxin Production by an emerging strain of Clostridium difficile associated with outbreaks of severe disease in North America and Europe. Lancet. 366:1079-1084.
Weese, J.S., Avery, B.P., Rousseau, J., Reid-Smith, R.J. (2009) Detection and enumeration of Clostridium difficile spores in retail beef and pork. Appl Environ Microbiol. 75:5009 5011.
Yamoudi, M., Mirlohi, M., Isfahani, B.N., Jalali, M., Esfandiari, Z., Hosseini, N.S. (2015) Isolation of toxigenic Clostridium difficile from ready-to-eat salads by multiplex polymerase chain reaction in Isfahan, Iran. Advanced Biomed Res. 4:87.