Bìa tạp chí


Prevalence of Clostridium perfringens and Clostridium difficile carrying cpa, tcdA toxin genes in raw meat and leafy vegetables

Published 03/31/2023

Article Details

How to Cite
Ta Thi Yen, Ninh Thi Hanh, Vu Thi Hai Ha, Pham Van Quan, Nguyen Thanh Trung. "Prevalence of Clostridium perfringens and Clostridium difficile carrying cpa, tcdA toxin genes in raw meat and leafy vegetables". Vietnam Journal of Food Control. vol. 6, no. 1, pp. 83-99, 2023

Main Article Content


The genus Clostridium is a Gram-positive, spore-forming bacterium. Most species of Clostridium spp. usually are found in soil, water, dead plants, animal carcasses, and play an important role in the decomposition of substances in nature. Raw meat and fresh vegetable are more likely to be contaminated with spores or vegetative cells of the genus Clostridium. The study results showed that the percentage of samples contaminated with raw meat and leafy vegetables collected at markets in Dong Da, Ha Dong and Hoang Mai districts infected with C. perfringens was 26.67% (n = 150) and 26.82% (n = 220). The percentage of raw meat and leafy vegetable samples contaminated with C. difficile was very low with 6.00% (n = 150) and 0.9% (n = 220), respectively. Gene amplification reactions revealed the presence of cpa toxin gene in all C. perfringens strains and detected 02 C. difficile strains carrying the tcdA gene.


C. perfringens and C. difficile, cpa gene, tcdA gene.


[1]. World Bank, “Vietnam food safety risks management : challenges and opportunities,” Washington, D.C. : World Bank Group, 2017.
[2]. General Statistics Office, “Statistical Yearbook,” 2019 (in Vietnamese). Address: https://www.gso.gov.vn/wp-content/uploads/2020/09/Nien-giam-thong-keday-du-2019.pdf.
[3]. Directorate for Standards, Metrology and Quality, “Vietnamese Standard TCVN 4991:2005 (ISO 7937:2004) on Microorganisms in food and animal feed - Quantitative method of Clostridium perfringens on agar plate - Counting technique colonies,” 2005 (in Vietnamese).
[4]. Directorate for Standards, Metrology and Quality, “Vietnam Standard TCVN 6404:2016 (ISO 7218: 2013) Microbiology of Food and Feed - General Requirements and Guidelines for Microbiological Testing,” 2016 (in Vietnamese).
[5]. E. Catherine, B. Burghoffer and F. Barbut, “Contamination of ready-to-eat raw vegetables with Clostridium difficile in France,” Journal of Medical Microbiology, vol. 62, pp. 1435-1438, 2013.
[6]. E. Catherine, B. Burghoffer, and F. Barbut, “Evaluation of the Chromogenic Agar chromID C. difficile,” Journal of Clinical Microbiology, vol. 51, no. 3, pp. 1002-1004, 2013.
[7]. Perry J. D., “A decade of development of chromogenic culture media for clinical microbiology in an era of molecular diagnostics,” Clinical Microbiology Review, 30, pp.449-479, 2017.
[8]. Vitek MS, “Manual Instruction,” 2017.
[9]. M. Mudassar, Z. Iqbal, A. Siddique, S. Liao, M. K. Farooq Salamat, N. Qi, A. M. Din, and M. Sun, “Prevalence, Genotypic and Phenotypic Characterization and Antibiotic Resistance Profile of Clostridium perfringens Type A and D Isolated from Feces of Sheep (Ovis aries) and Goats (Capra hircus) in Punjab, Pakistan,” Toxins, vol. 12, no. 657, 2020.
[10]. L. Lemee, A. Dhalluin, S. Testelin, M-A. Mattrat, K. Maillard, J-F. Lemeland, and J-L. Pons, “Multiplex PCR targeting tpi (triose phosphate isomerase), tcdA (Toxin A), and tcdB (Toxin B) genes for toxigenic culture of Clostridium difficile,” Journal of Clinical Microbiololgy, vol. 42, no. 12, pp. 5710-5714, 2004.
[11]. L. M. Wijnands, A. van der Mey - Florijn, E. D. - van Asch, “Clostridium perfringens associated food borne disease: Final report,” National Institute for Public Health and the Environment, Ministry of Health, Welfare and Sport, the Netherland, 2011.
[12]. H. Wen-Si, H. Kim, and O. K. Koo, “Molecular genotyping, biofilm formation and antibiotic resistance of enterotoxigenic Clostridium perfringens isolated from meat supplied to school cafeterias in South Korea,” Anaerobe, 2018.
[13]. M. Bendary Mahmoud, M. I. Abd El-Hamid, R. M. El-Tarabili, A. A. Hefny, R. M. Algendy, N. A. Elzohairy, M. M. Ghoneim, M. M. Al-Sanea, M. H. Nahari, and W. H. Moustafa, “Clostridium perfringens Associated with Foodborne Infections of Animal Origins: Insights into Prevalence, Antimicrobial Resistance, Toxin Genes Profiles, and Toxinotypes,” Biology, vol. 11, no. 551, 2022.
[14]. M. Yasuhiro, K. Miyamoto, I. Kaneko-Hirano, K. Fujiuchi, and S. Akimoto, “Prevalence and Characterization of Enterotoxin Gene-Carrying Clostridium perfringens Isolates from Retail Meat Products in Japan,” Applied and Environmental Microbiology, vol/. 74, no. 17, pp. 5366-5372, 2008.
[15]. H. S. Guran and G. Oksuztepe, “Detection and typing of Clostridium perfringens from retailchicken meat parts,” Letters in Applied Microbiology, vol. 57, pp. 77-82, 2013.
[16]. E. C. Emelda, F. O. Nwaokorie, A. O. Coker, M. J. Avila-Campos, R. L. Solis, L. A. Llanco, and F. T. Ogunsola, “Detection of toxigenic Clostridium perfringens and Clostridium botulinum from food sold in Lagos, Nigeria,” Anaerobe, vol. 42, pp. 176-181, 2016.
[17]. J. Yanfen, Y. Ma, Q. Liu, T. Li, Y. Li, K. Guo, and Y. Zhang, “Tracing Clostridium perfringens strains from beef processing of slaughter house by pulsed-field gel electrophoresis, and the distribution and toxinotype of isolates in Shaanxi province, China,” Food Microbiology, vol. 101, pp. 1-10, 2022.
[18]. A. Issimov, T. Baibatyrov, A. Tayeva, S. Kenenbay, S. Abzhanova, G. Shambulova, G. Kuzembayeva, M. Kozhakhiyeva, I. Brel-Kisseleva, O. Safronova, B. Lyailya, Y. Gulzhan, B. Kainar, K. Alma, and F. A. Uzal, “Prevalence of Clostridium perfringens and Detection of its Toxins in Meat Products in Selected Areas of West Kazakhstan,” Agriculture, vol. 12, no. 1357, pp. 1-6, 2022.
[19]. A. Masoumeh, B. Nadalian, H. Alavifard, S. N. Panirani, S. M. Vand Bonab, F. Azimirad, F. Gholami, P. Jabbari, A. Yadegar, L. Busani, H. A. Aghdaei, M. R. Zali, “Microbiological survey and occurrence of bacterial foodborne pathogens in raw and ready-to-eat green leafy vegetables marketed in Tehran, Iran,” International Journal of Hygiene and Environmental Health, vol. 237, no. 113824, pp. 1-9, 2021.
[20]. S. Ghorchian, M. Douraghi, A. Rahimiforoushani, M. M. Soltan Dallal, “Isolation and identification of cpe-positive Clostridium perfringens in bulk and packed dehydrated vegetables,” Razi Journal of Medical Science, vol. 26, no. 8, pp. 23-30, 2021.
[21]. H. Atsushi, H. Suzuki, and K. Oonaka, “Prevalence of cpe-positive Clostridium perfringens in surface-attached soil of commercially available potatoes and its significance as a potential source of food poisoning,” Anaerobe, vol. 79, no. 102687, 2023.
[22]. A. A. Mahamat, “Characterization of Clostridium perfringens strains for the investigation of food poisoning outbreaks in France,” Foodborn Pathogens and Whole Genome Sequencing Joint Conference, 26-28 March, 2019.
[23]. L. Tingting, H. Bian, Z. Sun, X. Wang, F. Liu, and D. Wang, "Inactivation of clostridium perfringens C1 spores by the combination of mild heat and lactic acid," Foods, vol. 11, no. 23, 2022.
[24]. K. T. Onur, T. Uyanik, S. Kanat, Ö. Çadirci, and A. Gücükoğlu, “Detection of Clostridium perfringens and determination of enterotoxin genes (cpa and cpe) in traditional turkish chicken doner kebab,” Ankara Univ Vet Fak Derg, no. 69, pp. 117-122, 2020.
[25]. L. Yuan-Tong and R. Labbe, “Enterotoxigenicity and Genetic Relatedness of Clostridium perfringens Isolates from Retail Foods in the United States,” Applied And Environmental Microbiology, vol. 69, no. 3, pp. 1642-1646, 2003.
[26]. Tham Thi Nguyen, Hung Vu-Khac and Tan Duc Nguyen, “Isolation and characterization of Clostridium perfringens strains isolated from ostriches (Struthio camelus) in Vietnam,” Veterinary World, vol. 13, no. 8, pp. 1679-1684, 2020.
[27]. C. Rodriguez, B. Taminiau, J. Van Broeck, M. Delme´e, and G. Daube, “Clostridium difficile in Food and Animals: A Comprehensive Review,” Advances in Experimental Medicine and Biology, 2016.
[28]. A. Rodriguez-Palacios, C. Pickworth, S. Loerch, and JT. LeJeune, “Transient fecal shedding and limited animal-to-animal transmission of Clostridium difficile by naturally infected finishing feedlot cattle,” Applied and Environmental Microbiology, vol. 77, pp. 3391-3397, 2011.
[29]. P. Hawken, J. S. Weese, R. Friendship, and K. Warriner, “Carriage and dissemination of Clostridium difficile and methicillin resistant Staphylococcus aureus in pork processing,” Food Control, no. 31, pp. 433-437, 2013.
[30]. S. Bouttier, M-C. Barc, B. Felix, S.Lambert, A. Collignon, and F. Barbut, “Clostridium difficile in Ground Meat, France,” Emerging Infectious Diseases, vol. 16, no. 4, 2010.
[31]. A. Rodriguez-Palacios, S. Borgmann, T. R. Kline, “Clostridium difficile in foods and animals: history and measures to reduce exposure,” Animal Health Research Reviews, vol. 14, pp. 11-29, 2013.
[32]. R. Ebrahim, M. Jalali, and J. S.Weese, “Prevalence of Clostridium difficile in raw beef, cow, sheep, goat, camel and buffalo meat in Iran,” BMC Public Health 14, 2014.
[33]. K. Muratoglu, E. Akkaya1, H. Hampikyan, E. Baris Bingol, O. Cetin, and H. Colak, “Detection, Characterization and Antibiotic Susceptibility of Clostridioides (Clostridium) difficile in Meat Products,” Food Science and Animal. Resources, vol. 40, no. 4, pp.578-587, 2020.
[34]. K. Malihe, S. Jalilian, A. Alvandi, and R.Abiri, “Prevalence of Clostridium difficile and its toxigenic genotype in beef samples in west of Iran,” IJM Iranian Journal of Microbiology, vol. 9, no. 3, pp. 169-173, 2017.
[35]. S. Di Bella, P. Ascenzi, S. Siarakas, N. Petrosillo, and A. di Masi, “Clostridium difficile Toxins A and B: Insights into Pathogenic Properties and Extraintestinal Effects,” Toxins, vol. 8, no. 134, 2016.