Incidence of Clostridium perfringens in Intestinal Contents of Domestic Livestock Detected by PCR


Incidence of Clostridium perfringens in Intestinal Contents of Domestic Livestock Detected by PCR


Asghar Arshi1, Sajad Nikkhah2, Hamidreza Kabiri3, Arman Akbarpour2, Rassoul Hashemzehi4, Behnaz Mansouri2, Ayse Kilic5, Esmaeil Mahmoudi2*

1. Young Researchers and Elite Club, Najafabad Branch, Islamic Azad University, Najafabad, Iran
2. Biotechnology Research Center, Islamic Azad University, Shahrekord Branch, Shahrekord, Iran
3. Young Researchers and Elite Club, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran
4. Department of Genetics, Fars Science & Research Branch, Islamic Azad University, Shiraz, Iran
5. Sivrice Vocational High School, University of Firat, 23119 Elazig-Turkey


International Journal of Animal ResearchClostridium perfringens (C. perfringens) is a Gram-positive anaerobic spore-forming bacterium that is widespread in environmental soil and sewage, as well as in animal intestines. C. perfringens is an important pathogen in both human and veterinary medicine. The incidence and numbers of C. perfringens in the intestinal contents of 100 cattle, 86 turkey and 177 chickens from September 2015 to April 2016 were determined. C. perfringens was found in 26%, 22% and 40% of intestinal contents of cattle, turkey and chickens, respectively. In this research samples were tested for isolation of C. perfringens by culturing and biochemical method and then they were confirmed by the polymerase chain reaction (PCR) methods. We suggest that PCR assay could be a replacement of the culture method for quantifying C. perfringens in the intestinal tracts. These research helping us to establish the role of each C. perfringens toxin in animal disease, to investigate the in vivo mechanism of action of these toxins, and to develop more effective vaccines against diseases produced by these microorganisms.


Keywords: Clostridium perfringens, Cattle, Turkey, Chickens, PCR.

Free Full-text PDF


How to cite this article:
Asghar Arshi et al., Incidence of Clostridium perfringens in Intestinal Contents of Domestic Livestock Detected by PCR. International Journal of Animal Research, 2017; 1:9. DOI:10.28933/ijar-2017-09-2801


References:
1. Obrien DK, Melville SB. The anaerobic pathogen Clostridium perfringens can escape the phagosome of macrophages under aerobic conditions. Cellular microbiology. 2000; 2: 505-19.
2. Jabbari AR, Tekyei F, Esmaeilizad M, Pilehchian Langroudi R. Occurrence of Beta2 toxigenic Clostridium perfringens isolates with different toxin types in Iran. Archives of Razi. 2012; 67: 133-7.
3. Shimizu T, Ohtani K, Hirakawa H, Ohshima K, Yamashita A, Shiba T, et al. Complete genome sequence of Clostridium perfringens, an anaerobic flesh-eater. Proceedings of the National Academy of Sciences. 2002; 99: 996-1001.
4. Knowles TG, Kestin SC, Haslam SM, Brown SN, Green LE, Butterworth A, et al. Leg disorders in broiler chickens: prevalence, risk factors and prevention. PLoS One. 2008; 3: e1545.
5. Welch WH, Nuttall GHF. A gas producing bacillus (Bacillus aerogenes capsulatus, nov, sepc.) capable of rapid development in the blood vessels after death. Bulletin of the Johns Hopkins Hospital. 1892; 3: 81-91.
6. Yoo HS, Lee SU, Park KY, Park YH. Molecular typing and epidemiological survey of prevalence of Clostridium perfringens types by multiplex PCR. Journal of clinical microbiology. 1997; 35: 228-32.
7. Drew MD, Syed NA, Goldade BG, Laarveld B, Van Kessel AG. Effects of dietary protein source and level on intestinal populations of Clostridium perfringens in broiler chickens. Poultry science. 2004; 83: 414-20.
8. Sartory DP, Field M, Curbishley SM, Pritchard AM. Evaluation of two media for the membrane filtration enumeration of Clostridium perfringens from water. Letters in Applied Microbiology. 1998; 27: 323-27.
9. Wells CL, Wilkins TD. Clostridia (4th edition): sporeforming anaerobic bacilli. Medical microbiology. 1996.
10. Asha NJ, Wilcox MH. Laboratory diagnosis of Clostridium perfringens antibiotic-associated diarrhea. Journal of medical microbiology. 2002; 51: 891-94.
11. Rood JI. Virulence genes of Clostridium perfringens. Annual Reviews in Microbiology. 1998; 52: 333-60.
12. Effat MM, Abdallah YA, Soheir MF, Rady MM. Characterization of Clostridium perfringens field isolates, implicated in necrotic entritis outbreaks on private broiler farms in Cairo, by multiplex PCR. African Journal of Microbiology Research. 2007; 29-32.
13. Immerseel FV, Buck JD, Pasmans F, Huyghebaert G, Haesebrouck F, Ducatelle R. Clostridium perfringens in poultry: an emerging threat for animal and public health. Avian Pathology. 2004; 33: 537-49.
14. Lin YT, Labbe R. Enterotoxigenicity and genetic relatedness of Clostridium perfringens isolates from retail foods in the United States. Appl Environ Microbiol. 2003; 69: 1642-6.
15. Williamson ED, Titball RW. A genetically engineered vaccine against the alpha-toxin of Clostridium perfringens protects mice against experimental gas gangrene. Vaccine. 1993; 11: 1253-8.
16. Laura Z, Ossiprandi MC. Molecular toxinotyping of clostridium perfringens cattle isolates by multiplex PCR. Annali Della Facoltà di Medicina Veterinaria di Parma. 2010; 71-80.
17. Smedley JG, Fisher DJ, Sayeed S, Chakrabarti G, McClane BA. The enteric toxins of Clostridium perfringens. Reviews of Physiology, Biochemistry and Pharmacology. 2004; 152: 183-204.
18. Sawires YS, Songer JG. Clostridium perfringens: insight into virulence evolution and population structure. Anaerobe. 2006; 12: 23-43.
19. Tantawiwat S, Tansuphasiri U, Wongwit W, Wongchotigul V, Kitayaporn D. Development of multiplex PCR for the detection of total coliform bacteria for Escherichia coli and Clostridium perfringens in drinking water. The Southeast Asian Journal of Tropical Medicine and Public Health. 2005; 36: 162-9.
20. Allaart JG, van Asten AJAM, Vernooij JCM, Gröne A. Effect of Lactobacillus fermentum on beta2 toxin production by Clostridium perfringens. Applied and environmental microbiology. 2011; 77: 4406-11.
21. Brynestad S, Granum PE. Clostridium perfringens and foodborne infections. International Journal of Food Microbiology. 2002; 74: 195-202.
22. Lindström M, Heikinheimo A, Lahti P, Korkeala H. Novel insights into the epidemiology of Clostridium perfringens type A food poisoning. Food Microbiology. 2011; 28: 192-8.
23. Mersmann HJ. Beta-adrenergic receptor modulation of adipocyte metabolism and growth. Journal of Animal Science. 2002; 80: E24-E29.
24. Rahimi S, Kathariou S, Grimes JL, Siletzky RM. Effect of direct-fed microbials on performance and Clostridium perfringens colonization of turkey poults. Poultry Science. 2011; 90: 2656-62.
25. Songer JG. Clostridia as agents of zoonotic disease. Veterinary Microbiology. 2010; 140: 399-404.
26. Johnson EA, Summanen P, Finegold SM. Clostridium. Manual of Clinical Microbiology. 2007; 9: 889-910.
27. Collier CT, Van Der Klis JD, Deplancke B, Anderson DB, Gaskins HR. Effects of tylosin on bacterial mucolysis, Clostridium perfringens colonization, and intestinal barrier function in a chick model of necrotic enteritis. Antimicrobial agents and chemotherapy. 2003; 47: 3311-7.
28. Langroudi RP, Pour KA, Shamsara M, Ghorashi SA. Synthetic construct of Clostridium perfringens epsilon-beta fusion protein gene. International Conference on Biotechnology and Environment Management. 2011; 18: 126-131.
29. Tolooe A, Shojadoost B, Peighambari SM. Molecular detection and characterization of cpb2 gene in Clostridium perfringens isolates from healthy and diseased chickens. Journal of Venomous Animals and Toxins including Tropical Diseases. 2011; 17: 59-65.
30. Wnek AP, Strouse RJ, McClane BA. Production and characterization of monoclonal antibodies against Clostridium perfringens type A enterotoxin. Infection and immunity. 1985; 50: 442-8.
31. Uzal FA, Vidal JE, McClane BA, Gurjar AA. Clostridium perfringens toxins involved in mammalian veterinary diseases. Open Toxinology Journal. 2010; 3: 24-42.
32. Songer JG. Clostridial diseases of small ruminants. Veterinary Research. 1998; 29: 219-32.
33. Jabbari AR, Afshari Far AR, Esmaelizad M, Pilehchian Langroudi R, Abdolmohammadi Khiav L. Molecular typing of toxigenic Clostridum perfringens isolated from sheep in Iran. Archives of Razi. 2011; 66: 81-86.
34. Asmuth DM, Olson RD, Hackett SP, Bryant AE, Tweten RK, Tso JY, et al. Effects of Clostridium perfringens recombinant and crude phospholipase C and theta-toxin on rabbit hemodynamic parameters. The Journal of Infectious Diseases. 1995; 172: 1317-23.
35. Stevens DL, Troyer BE, Merrick DT, Mitten JE, Olson RD. Lethal effects and cardiovascular effects of purified alpha- and theta-toxins from Clostridium perfringens. The Journal of Infectious Diseases. 1988; 157: 272-9.
36. Birdsall HH, Green DM, Trial J, Youker KA, Burns AR, MacKay CR, et al. Complement C5a, TGF-beta 1, and MCP-1, in sequence, induce migration of monocytes into ischemic canine myocardium within the first one to five hours after reperfusion, Circulation. 1997; 95: 684-92.
37. Gourmala NG, Buttini M, Limonta S, Sauter A, Boddeke HW. Differential and time-dependent expression of monocyte chemoattractant protein-1 mRNA by astrocytes and macrophages in rat brain: effects of ischemia and peripheral lipopolysaccharide administration. Journal of neuroimmunology. 1997; 74: 35-44.
38. Ivacko J, Szaflarski J, Malinak C, Flory C, Warren JS, Silverstein FS. Hypoxic-ischemic injury induces monocyte chemoattractant protein-1 expression in neonatal rat brain. Journal of Cerebral Blood Flow & Metabolism. 1997; 17: 759-70.
39. Stevens DL, Maier KA, Mitten JE. Effect of antibiotics on toxin production and viability of Clostridium perfringens. Antimicrobial agents and chemotherapy. 1987; 31: 213-18.
40. Fach P, Popoff MR. Detection of enterotoxigenic Clostridium perfringens in food and fecal samples with a duplex PCR and the slide latex agglutination test. Applied and environmental microbiology. 1997; 63: 4232-36.
41. Shandera WX, Tacket CO, Blake PA. Food poisoning due to Clostridium perfringens in the United States. The Journal of Infectious Diseases. 1983; 147: 167-70.
42. Heikinheimo A, Korkeala H. Multiplex PCR assay for toxinotyping Clostridium perfringens isolates obtained from Finnish broiler chickens. Letters in applied microbiology. 2005; 40: 407-11.
43. Baums CG, Schotte U, Amtsberg G, Goethe R. Diagnostic multiplex PCR for toxin genotyping of Clostridium perfringens isolates. Veterinary Microbiology. 2004; 100: 11-6.
44. Park JY, Kim S, Oh JY, Kim HR, Jang I, Lee HS, et al. Characterization of Clostridium perfringens isolates obtained from 2010 to 2012 from chickens with necrotic enteritis in Korea. Poult Sci. 2015; 94: 1158-64.
45. Goncuoglu EM, Ayaz ND, Ormanci FS, Hidebrandt G. Molecular typing of Clostridium perfringens isolated from turkey meat by multiplex PCR. Letters in Applied Microbiology. 2008; 47: 31-4.
46. Drouin P. Retour en force de l’entérite nécrotique. Filieres Avicoles. 1999; 78-9.
47. Bueschel DM, Jost BH, Billington SJ, Trinh HT, Songer JG. Prevalence of cpb2, encoding beta2 toxin, in Clostridium perfringens filed isolates: correlation of genotype with phenotype. Veterinary Microbiology. 2003; 94: 121-9.
48. Zerbini L, Ossipranadi MC. Molecular typing of Clostridium perfringens isolated from dogs by toxin gene amplification. Annual of Veterinary Medicine Faculty Parma. 2009; 29: 115-28.
49. Ahsani MR, Mohammadabadi MR, Shamsaddini MB. Clostridium perfringens isolate typing by multiplex PCR. Journal of Venomous Animals and Toxins including Tropical Diseases. 2010; 16: 573-8.