Comparison of toxin production power in different types of Clostridium perfringens among sheep and goats isolates by ELISA

Document Type : Full Research Paper

Authors

1 Department of Microbiology, Kerman Branch, Islamic Azad University, Kerman, Iran

2 Department of Research and Technology, Kerman Branch, Razi Vaccine & Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Kerman, Iran

Abstract

Clostridium perfringens (C. perfringens) is one of the most common pathogens that produce an array of toxins that are secreted in the body which cause economic losses in the livestock industry. C. perfringens has four major toxins: alpha, beta, epsilon, and iota. A total of 84 isolates of C. perfringens from various types that originated from sheep and goats' isolates that were identified as C. perfringens by microbiological, biochemical, and toxin typed by PCR method were evaluated for production of alpha, beta, and epsilon toxin by ELISA method. Results showed that out of 84, 64 isolates showed the presence of at least one toxin in ELISA test, 31, and six isolates were positive for only alpha and only epsilon toxin, respectively, 17, six isolates show the presence of both alpha and epsilon and both alpha and beta toxins, respectively furthermore four isolates positive for the presence of all three toxins, meanwhile, 20 C. perfringens isolates didn’t show any of the toxins. The results were compared with the results of toxinotyping by PCR method. The frequency and distribution of toxins in different strains were evaluated, so type C isolates showed the most positive cases of alpha and beta toxin and type B isolates showed the most positive cases of epsilon toxin, which could be related to the pathogenicity of these types in the isolates studied

Keywords

References

1- Abdolmohammadi Khiav, L., A. Paradise and A. Hagh Roosta. 2020. Designing of an Indirect ELISA method for the detection of Beta antitoxin of Clostridium perfringens Type C in Rabbit Serum. Veterinary Researches & Biological Products 33:21-30. 
2-Ahmadi Rahnemoon, A., S. Esmaeilzadeh, B. Mohammadian, M. Ghorbanpoor and A. Ghadrdan Mashhadi. 2020. Ovine enterotoxemia in Ahvaz region, pathological, bacteriological, serological and molecular studies. Iranian Veterinary Journal 16: 25-39.
3- Ahsani M., M. Shamsaddini Bafti, A. Esmailizadeh and M. Mohammadabadi. 2011. Genotyping of isolates of Clostridium perfringens from vaccinated and unvaccinated sheep. Small Ruminant Research 95: 65-69.
4- Albini, S., I. Brodard, A. Jaussi, N. Wollschläger, J. Frey, R. Miserez and C. Abril. 2008. Real-time multiplex PCR assays for reliable detection of Clostridium perfringens toxin genes in animal isolates. Veterinary Microbiology 127: 179-185.
5- Babe, T., B. Jafari and S. Bahmanpour. 2012. Typing of toxigenic isolates of Clostridium perfringens by ELISA in ostrich. African Journal of Microbiology Research 6: 1766-1769.
6- Baums, C. G., U. Schotte, G. Amtsberg and R. Goethe. 2004. Diagnostic multiplex PCR for toxin genotyping of Clostridium perfringens isolates. Veterinary Microbiology 100: 11-16.
7- Dürre, P. 2005. Handbook on clostridia. CRC press.
8- El Idrissi, A. H. and G. E. Ward. 1992. Evaluation of enzyme-linked immunosorbent assay for diagnosis of Clostridium perfringens enterotoxemias. Veterinary Microbiology 31: 389-396.
9- Ezatkhah, M., M. Alimolaei, M. Amini and M. Shamsaddini Bafti. 2016. Typing toxigenic Clostridium perfringens strains from the ruminants of Yazd province by multiplex polymerase chain reaction. Int J Enteric Pathog 4: 1-4.
10- Forti, K., L. Ferroni, M. Pellegrini, D. Cruciani, A. De Giuseppe, S. Crotti, P. Papa, C. Maresca, G. Severi and M. L. Marenzoni. 2020. Molecular characterization of Clostridium perfringens strains isolated in Italy. Toxins 12: 650.
11- Goekce, H. I., O. Genç, M. Soezmen and G. Gökçe. 2007. Determination of Clostridium perfringens toxin-types in sheep with suspected enterotoxemia in Kars Province, Turkey. Turkish Journal of Veterinary and Animal Sciences 31: 355-360.
12- Hadimli, H. H., O. Erganiş, Z. Sayin and Z. Aras. 2012. Toxinotyping of Clostridium perfringens isolates by ELISA and PCR from lambs suspected of enterotoxemia. Turkish Journal of Veterinary and Animal Sciences 36: 409-415.
13- Hayati, M. and Y. Tahamtan. 2020. Toxin typing of Clostridium perfringens associated with enterotoxaemia cases of sheep and healthy group in Fars province by PCR and ELISA methods. Archives of Razi Institute.76(3).
14- Jabbari, A., S.A.R. Afshari, M. Esmaelizad, R. Pilehchian Langroudi, M. Moosawi Shooshtari and L. Abdolmohammadi. Khiav. 2011. Molecular typing of toxigenic Clostridum perfringens isolated from sheep in Iran. Archives of Razi Institute 66: 81-86.
15- Karimabadizadeh, A. R. and M. Shamsaddini Bafti. 2020. Evaluation of toxinogenesis of Clostridium perfringens type D isolates in three kinds of culture media. Veterinary Researches & Biological Products 132: 2-12
16- Kiu, R. and L. J. Hall. 2018. An update on the human and animal enteric pathogen Clostridium perfringens. Emerging Microbes & Infections 7: 1-15.
17- Mehdizadeh Gohari, I., M. A. Navarro, J. Li, A. Shrestha, F. Uzal and B. A. McClane. 2021. Pathogenicity and virulence of Clostridium perfringens. Virulence 12: 723-753.
18- Møller, K. and P. Ahrens. 1996. Comparison of toxicity neutralization-, ELISA-and PCR tests for typing of Clostridium perfringens and detection of the enterotoxin gene by PCR. Anaerobe 2: 103-110. 19- Nagahama, M., K. Kobayashi, S. Ochi and J. Sakurai. 1991. Enzyme-linked immunosorbent assay for rapid detection of toxins from Clostridium perfringens. FEMS Microbiology Letters 84: 41-44.
20- Paradise, A. and L. Abdolmohammadi Khiav. 2020. Evaluation of epsilon antitoxin of Clostridium Perfringens type D in the rabbit serum by indirect ELISA. Veterinary Researches & Biological Products 33(3): 17-30.
21- Quinn, P. J., B. K. Markey, F. C. Leonard, P. Hartigan, S. Fanning and E. Fitzpatrick. 2011. Veterinary microbiology and microbial disease. John Wiley & Sons.
22- Rood, J. I., V. Adams, J. Lacey, D. Lyras, B. A. McClane, S. B. Melville, R. J. Moore, M. R. Popoff, M. R. Sarker and J. G. Songer. 2018. Expansion of the Clostridium perfringens toxin-based typing scheme. Anaerobe 53: 5-10.
23- Sawires, Y. S. and J. G. Songer. 2006. Clostridium perfringens: insight into virulence evolution and population structure. Anaerobe 12: 23-43.
24- Sepehrifar, H., R. Pilehchian Langroudi, S. Ataei and A. Haddadi. 2021. Evaluation and comparison of Clostridium epsilon-alpha fusion gene expression using different commercial expression vector. Archives of Razi Institute 76: 7-16.
25- Shamsaddini Bafti, M., M. Ezzatkhah, R. Pilehchian, A. R. Jabbari and M. Hayati. 2016. Isolation and identification of types of Clostridium perfringens in the south of the country. Agricultural Research, Education and Extension Organization (AREEO).
26- Songer, J. G. 1997. Molecular and immunological methods for the diagnosis of clostridial diseases. The Clostridia. Academic Press 491-503. 
27- Songer, J. G. and R. R. Meer. 1996. Genotyping of Clostridium perfringens by polymerase chain reaction is a useful adjunct to
 diagnosis of Clostridial enteric disease in animals. Anaerobe 2: 197-203.
28- Uzal, F. A., J. C. Freedman, A. Shrestha, J. R. Theoret, J. Garcia, M. M. Awad, V. Adams, R. J. Moore, J. I. Rood and B. A. McClane. 2014. Towards an understanding of the role of Clostridium perfringens toxins in human and animal disease. Future Mmicrobiology 9: 361-377.
29- Uzal, F. A., W. Kelly, R. Thomas, M. Hornitzky and F. Galea. 2003. Comparison of four techniques for the detection of Clostridium perfringens type D epsilon toxin in intestinal contents and other body fluids of sheep and goats. Journal of Veterinary Diagnostic Iinvestigation 15: 94-99.
30- Van Asten, A. J., C. W. Van Der Wiel, G. Nikolaou, D. J. Houwers and A. Gröne. 2009. A multiplex PCR for toxin typing of Clostridium perfringens isolates. Veterinary Microbiology (Amsterdam) 136: 411-412.
31- Wu, J., W. Zhang, B. Xie, M. Wu, X. Tong, J. Kalpoe and D. Zhang. 2009. Detection and Toxin Typing of Clostridium perfringens in Formalin-Fixed, Paraffin-Embedded Tissue Samples by PCR. Journal of Clinical Microbiology 47(3): 807-810.
32- Yadegar, F., P. Nakhaei, G. Hashemtabari, G. Kalidari, M. Rashtibaf and J. Razmyar. 2018. Major and minor toxins of Clostridium perfringens isolated from healthy and diseased sheep. Small Ruminant Research 168: 1-5.
33- Zandi, E., M. Mohammadabadi, M. Ezatkhah and A. Esmailizadeh. 2014. Typing of toxigenic isolates of Clostridium perfringens by multiplex PCR in ostrich. Iranian Journal of Applied Animal Science 4: 795-801.
 
Veterinary Research & Biological Products
Volume 35, Issue 3 - Serial Number 3
September 2022
Pages 18-28

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History

  • Receive Date: 05 September 2021
  • Revise Date: 23 October 2021
  • Accept Date: 10 November 2021

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