Isolation and identification of Clostridium perfringens and its toxins from mutton in Lahore City, Punjab, Pakistan


Food-borne poisoning
Food safety
Multiplex PCR
Sheep and goat meet

How to Cite

Tul Abiha, S., Azeem, S., Ahmad Anjum, A. ., & Hassan Mushtaq, M. . (2023). Isolation and identification of Clostridium perfringens and its toxins from mutton in Lahore City, Punjab, Pakistan. Veterinaria Italiana, 59(4). https://doi.org/10.12834/VetIt.2926.20055.3


The present study was aimed to detect C. perfringens and identify its toxins in mutton samples collected from Lahore City in the Punjab Province of Pakistan. A total of 40 samples of minced and non‑minced mutton were collected from local butcher and retail shops representing four areas of the city. The samples were subjected to ELISA for the detection of C. perfringens alpha, beta and epsilon toxins. The samples were simultaneously processed for bacterial isolation. The isolates were confirmed both by biochemical testing and a multiplex PCR targeting alpha, beta and epsilon toxin genes of C. perfringens. While 10% (4/40) of the samples were positive for C. perfringens alpha toxins, 17.5% (7/40) of the samples were positive for the alpha toxin gene. The present study indicated that the samples collected from the local butcher shops were contaminated with C. perfringens and its toxins. Interestingly, no such contamination was detected in any of the samples collected from retail meat shops. In conclusion, improper hygienic conditions at butcher shops could lead to the contamination of mutton with C. perfringens and its toxins.



Bendary M.M., Abd El-Hamid M.I., El-Tarabili R.M., Hefny A.A., Algendy R.M., Elzohairy N.A., Ghoneim M.M., Al-Sanea M.M., Nahari M.H. & Moustafa W.H. 2022. Clostridium perfringens associated with foodborne infections of animal origins: insights into prevalence, antimicrobial resistance, toxin genes profiles, and toxinotypes. Biology 11, 551.

Brynestad S. & Granum P.E. 2002. Clostridium perfringens and foodborne infections. Int J Food Microbiol 74,195-202.

European Food Safety Authority. 2005. Opinion of the Scientific Panel on Biological Hazards on a request from the Commission related to Clostridium spp in foodstuffs. The EFSA Journal 199, 1–65.

Fukushima H., Hoshina K., Nakamura R. & Ito, Y. 1987. Raw beef, pork and chicken in Japan contaminated with Salmonella sp., Campylobacter sp., Yersinia enterocolitica, and Clostridium perfringens- a comparative study. Zentralblatt fur Bakteriologie, Mikrobiologie und Hygiene. Serie B, Umwelthygiene, Krankenhaushygiene, Arbeitshygiene, Praventive Medizin 184, 60-70.

Gohari I.M., A Navarro M., Li J., Shrestha A., Uzal F., & A McClane B. 2021. Pathogenicity and virulence of Clostridium perfringens. Virulence 12, 723-753.

Grass J.E., Gould L.H. & Mahon B.E. 2013. Epidemiology of foodborne disease outbreaks caused by Clostridium perfringens, United States, 1998–2010. Foodborne Pathog Dis 10, 131-136.

Hall H.E., & Angelotti R. 1965. Clostridium perfringens in meat and meat products. Appl Microbiol 13, 352-357.

Hamza D., Dorgham S.M., Elhariri M., Elhelw R., Ismael E. 2018. New insight of apparently healthy animals as a potential reservoir for Clostridium perfringens: a public health implication. J Vet Res 62, 457-462.

Hassani S., Pakbin B., Brück W.M., Mahmoudi R., & Mousavi S. 2022. Prevalence, antibiotic resistance, toxin-typing and genotyping of Clostridium perfringens in raw beef meats obtained from Qazvin City, Iran. Antibiotics (Basel) 11, 340.

Kamber U., Gokce H.I. & Elmali M. 2007. Clostridium perfringens and its toxins in minced meat from Kars, Turkey. Food Addit Contam 24, 673-678.

Khan M., Nazir J., Anjum A.A., Ahmad M.U., Nawaz M. & Shabbir M.Z. 2015. Toxinotyping and antimicrobial susceptibility of enterotoxigenic Clostridium perfringens isolates from mutton, beef and chicken meat. J Food Sci Technol 52, 5323-5328.

Khan M.A., Khan S.B., Khan I., Sultan A., Khan R., Shahid M., Aqib I., Khan S., Khan A. & Hassan M. 2020. The prevalence of Clostridium perfringens in retail meat of Mardan, Pakistan. Turkish J Vet Anim Sci 44, 618-623.

Kiu R. & Hall L.J. 2018. An update on the human and animal enteric pathogen Clostridium perfringens. Emerg Microbes Infect 7,141.

Leung V.H., Phan Q., Costa C.E., Nishimura C., Pung K., Horn L. & Sosa L. 2017. Notes from the Field: Clostridium perfringens Outbreak at a Catered Lunch - Connecticut, September 2016. MMWR Morb Mortal Wkly Rep 66, 940-941.

Magsi H., Randhawa A.A. & Shah A.H. 2021. Halal meat production in Pakistan: status and prospects. J Islam Market 12, 941-950.

Mellou K., Kyritsi M., Chrysostomou A., Sideroglou T., Georgakopoulou T. & Hadjichristodoulou C. 2019. Clostridium perfringens foodborne outbreak during an athletic event in Northern Greece, June 2019. Int J Environ Res Public Health. 16, 3967.

McClane B.A., Robertson S.L. & Li J. 2013. Clostridium perfringens In Food Microbiology, eds M. Doyle and R. Buchanan (Washington, DC: ASM Press), 2013; 465–489.

Miki Y., Miyamoto K., Kaneko-Hirano I., Fujiuchi K., & Akimoto S. 2008. Prevalence and characterization of enterotoxin gene-carrying Clostridium perfringens isolates from retail meat products in Japan. Appl Environ Microbiol 74, 5366-5372.

Miwa N., Nishina T., Kubo S. & Honda H. 1997. Most probable numbers of enterotoxigenic Clostridium perfringens in intestinal contents of domestic livestock detected by nested PCR. Journal Vet Med Sci 59, 557-560.

Rood J.I., Adams V., Lacey J., Lyras D., McClane B.A., Melville S.B., Moore R.J., Popoff M.R. Sarker M.R., Songer J.G., Uzal F.A., Van Immerseel F. 2018. Expansion of the Clostridium perfringens toxin-based typing scheme. Anaerobe 53, 5-10.

Taormina P.J. & Dorsa W.J. 2004. Growth potential of Clostridium perfringens during cooling of cooked meats. J Food Protection 67, 1537-1547.

van Asten A.J., van der Wiel C.W., Nikolaou G., Houwers D.J., Gröne A. 2009. A multiplex PCR for toxin typing of Clostridium perfringens isolates. Vet Microbiol 136, 411–412.

Wen Q., McClane B.A. 2004. Detection of enterotoxigenic Clostridium perfringens type A isolates in American retail foods. Appl Environ Microbiol 70, 2685-2691.

Yadav J.P., Kaur S., Dhaka P., Vijay D., Bedi J.S. 2022. Prevalence, molecular characterization, and antimicrobial resistance profile of Clostridium perfringens from India: A scoping review. Anaerobe 77, 102639.

Yibar A, Cetin E, Ata Z, Erkose E, Tayar M. 2018. Clostridium perfringens contamination in retail meat and meat-based products in Bursa, Turkey. Foodborne Pathog Dis 15, 239-245.

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