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Research Article

volume 52 issue 1 (january 2018) : 111-115,   Doi: 10.18805/ijar.v0iOF.9129

Characterization of enterotoxin producing Clostridium perfringens isolated from foods of animal origin

S
Shahnaz Haque
R
Rajeev Kumar Sharma
N
Nagendra Nath Barman
L
Laishram Babita Devi
L
Luit Moni Barkalita
I
Iftikar Hussain
1Department of Microbiology, College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati-781022, Assam
Cite article:- Haque Shahnaz, Sharma Kumar Rajeev, Barman Nath Nagendra, Devi Babita Laishram, Barkalita Moni Luit, Hussain Iftikar (2017). Characterization of enterotoxin producing Clostridium perfringens isolated from foods of animal origin. Indian Journal of Animal Research. 52(1): 111-115. doi: 10.18805/ijar.v0iOF.9129.

ABSTRACT

Screening of 105 samples comprising of milk, meat and their products for isolation of Clostridium perfringens revealed a total of 39 (37.14%) samples to be positive for C. perfringens, yielding an equal number of isolates. Among the total isolates, 11 (20.0%) belonged to milk and milk products, of which 10 were isolated from raw cow’s milk. The remaining isolate was from paneer sample. Similarly, 28 (56.0%) C. perfringens isolates were recovered from meat and meat products. Majority (37) of the isolates belonged to toxin type A while two isolates recovered each from raw beef and chicken meats were of toxin type C. Among the 39 isolates of C. perfringens, only 5(12.82%) isolates could reveal the presence of cpe (enterotoxin) gene. The protein profile (SDS-PAGE) of both the crude and partially purified enterotoxins prepared from cpe positive C. perfringens isolates (type A and C) revealed a very little difference. Both the partially purified enterotoxins of type A and C of C. perfringens displayed almost similar type of cytopathic effects (CPE) and death of cells in Vero cell line.

REFERENCES

  1. Barett, N.J. (1989) Milk borne diseases in England and Wales in the 1980’s. J. Soc. Dairy. Tech., 42:4-6.
  2. Baums, C. G., Schotte, U., Amtsberg, G. and Goethe, R. (2004) Diagnostic multiplex PCR for toxin genotyping of C. perfringens isolates. Vet. Microbiol., 100:11-16
  3. Das, A.; Mazumdar, Y,; Dutta,B.K.; Shome, B.R,; Bujarbaruah, K.M. and Kumar, A. (2008). Clostridium perfringens type A from Broiler Chicken with Necrotic Enteritis. Int. J. Poultry Sci., 7:601-609.
  4. Laemmli, U.K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage. Nature, 227; 680-685.
  5. Mahony, D.E., Gilliat, E., Dawson,S, Stockdale, E and Lee, S.H.S. (1989) Vero cell for rapid detection of C. perfringens enterotoxins. Appl. Environ. Microbiol., 55:2141-2143.
  6. McClane, B.A. and Wnek, A.P. (1990) Studies of Clostridium perfringens enterotoxin action at different temperatures demonstrate a correlation between complex formation and cytotoxicity. Infect. And Immun., 58:3109-3115.
  7. McClane, B.A. (2007) Clostridium perfringens. In: Food microbiology: fundamentals and frontiers. Doyle M.P., Beauchat L.R., editors .Washington, DC: ASM Press; 2007. Pp423-44.
  8. Morera, J., Rodriguez, E. and Gamboa M.M. (1999). Determination of Clostridium perfringens in pork sausagesfrom the Metropolitan area of Costa Rica. Arch. Latinoam Nutr.,49:279-82.
  9. Moustafa, S.I. and Martin, E.H. (1993). Prevalence of Clostridium perfringens in bovine milk. Milchwissenschaft. 48:383-385.
  10. Norhan, A.K., Ammlar, A.M., Mona, A.M. and Hanaa, A.A. (2010) Studies on C. perfringens types A and D Enterotoxins and their sporulation.Innocult.
  11. Norinaga, M., Takashi, M., Katsuya T., Asako, K., Kazuhiro, O. and Miyamoto, H. (1998) Bacteriological investigation of an outbreak of Clostridium perfringens food poisoning caused by Japanese food without animal protein. Inter. J. of Food Microbiol .,49:103–106.
  12. Osman, K.M., Enbaawy. E.I., Ezzeldeen. N.A. and Hussain, H.M.G. (2009) Mastitis in dairy buffalo and cattle in Egypt due to Clostridium perfringens: prevalence, incidence, risk factors and costs. Rev. Sci. tech. Off. Int. Epiz., 28: 975-986.
  13. Piatti, R.M., Ikuno, A.A. and Baldassi, L. (2004). Detection of C. perfringens by polymerase chain reaction. J. Venom. Anim. Toxins incl. Trop. Dis., 10:154-160.
  14. Singh, R.V., Bhilegaonkar, K.N. and Agarwal, R.K. (2005). Studies on occurance and characterisations of Clostridium perfringens from select meats. J. Food Safety, 25: 146-156.
  15. Titball, R.W., Hunter, S.E.C., Martin, K.L., Morris, A.D., Shuttle worth, A.D., Rubdige, T., Anderson, D.W., Kelly, D.C. (1989) Molecular cloning and nucleotide sequence of the alpha toxin of Clostridium perfringens . Infect. Immun. 57:367-376.
  16. Uemura, T., Maekawa, T. and Sakaguchi, G. (1984). Biological Assay for C. perfringens enterotoxin with vero cells. Jpn. J. Vet. Sci., 46:715-720. 
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    Research Article

    volume 52 issue 1 (january 2018) : 111-115,   Doi: 10.18805/ijar.v0iOF.9129

    Characterization of enterotoxin producing Clostridium perfringens isolated from foods of animal origin

    S
    Shahnaz Haque
    R
    Rajeev Kumar Sharma
    N
    Nagendra Nath Barman
    L
    Laishram Babita Devi
    L
    Luit Moni Barkalita
    I
    Iftikar Hussain
    1Department of Microbiology, College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati-781022, Assam
    Cite article:- Haque Shahnaz, Sharma Kumar Rajeev, Barman Nath Nagendra, Devi Babita Laishram, Barkalita Moni Luit, Hussain Iftikar (2017). Characterization of enterotoxin producing Clostridium perfringens isolated from foods of animal origin. Indian Journal of Animal Research. 52(1): 111-115. doi: 10.18805/ijar.v0iOF.9129.

    ABSTRACT

    Screening of 105 samples comprising of milk, meat and their products for isolation of Clostridium perfringens revealed a total of 39 (37.14%) samples to be positive for C. perfringens, yielding an equal number of isolates. Among the total isolates, 11 (20.0%) belonged to milk and milk products, of which 10 were isolated from raw cow’s milk. The remaining isolate was from paneer sample. Similarly, 28 (56.0%) C. perfringens isolates were recovered from meat and meat products. Majority (37) of the isolates belonged to toxin type A while two isolates recovered each from raw beef and chicken meats were of toxin type C. Among the 39 isolates of C. perfringens, only 5(12.82%) isolates could reveal the presence of cpe (enterotoxin) gene. The protein profile (SDS-PAGE) of both the crude and partially purified enterotoxins prepared from cpe positive C. perfringens isolates (type A and C) revealed a very little difference. Both the partially purified enterotoxins of type A and C of C. perfringens displayed almost similar type of cytopathic effects (CPE) and death of cells in Vero cell line.

    REFERENCES

    1. Barett, N.J. (1989) Milk borne diseases in England and Wales in the 1980’s. J. Soc. Dairy. Tech., 42:4-6.
    2. Baums, C. G., Schotte, U., Amtsberg, G. and Goethe, R. (2004) Diagnostic multiplex PCR for toxin genotyping of C. perfringens isolates. Vet. Microbiol., 100:11-16
    3. Das, A.; Mazumdar, Y,; Dutta,B.K.; Shome, B.R,; Bujarbaruah, K.M. and Kumar, A. (2008). Clostridium perfringens type A from Broiler Chicken with Necrotic Enteritis. Int. J. Poultry Sci., 7:601-609.
    4. Laemmli, U.K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage. Nature, 227; 680-685.
    5. Mahony, D.E., Gilliat, E., Dawson,S, Stockdale, E and Lee, S.H.S. (1989) Vero cell for rapid detection of C. perfringens enterotoxins. Appl. Environ. Microbiol., 55:2141-2143.
    6. McClane, B.A. and Wnek, A.P. (1990) Studies of Clostridium perfringens enterotoxin action at different temperatures demonstrate a correlation between complex formation and cytotoxicity. Infect. And Immun., 58:3109-3115.
    7. McClane, B.A. (2007) Clostridium perfringens. In: Food microbiology: fundamentals and frontiers. Doyle M.P., Beauchat L.R., editors .Washington, DC: ASM Press; 2007. Pp423-44.
    8. Morera, J., Rodriguez, E. and Gamboa M.M. (1999). Determination of Clostridium perfringens in pork sausagesfrom the Metropolitan area of Costa Rica. Arch. Latinoam Nutr.,49:279-82.
    9. Moustafa, S.I. and Martin, E.H. (1993). Prevalence of Clostridium perfringens in bovine milk. Milchwissenschaft. 48:383-385.
    10. Norhan, A.K., Ammlar, A.M., Mona, A.M. and Hanaa, A.A. (2010) Studies on C. perfringens types A and D Enterotoxins and their sporulation.Innocult.
    11. Norinaga, M., Takashi, M., Katsuya T., Asako, K., Kazuhiro, O. and Miyamoto, H. (1998) Bacteriological investigation of an outbreak of Clostridium perfringens food poisoning caused by Japanese food without animal protein. Inter. J. of Food Microbiol .,49:103–106.
    12. Osman, K.M., Enbaawy. E.I., Ezzeldeen. N.A. and Hussain, H.M.G. (2009) Mastitis in dairy buffalo and cattle in Egypt due to Clostridium perfringens: prevalence, incidence, risk factors and costs. Rev. Sci. tech. Off. Int. Epiz., 28: 975-986.
    13. Piatti, R.M., Ikuno, A.A. and Baldassi, L. (2004). Detection of C. perfringens by polymerase chain reaction. J. Venom. Anim. Toxins incl. Trop. Dis., 10:154-160.
    14. Singh, R.V., Bhilegaonkar, K.N. and Agarwal, R.K. (2005). Studies on occurance and characterisations of Clostridium perfringens from select meats. J. Food Safety, 25: 146-156.
    15. Titball, R.W., Hunter, S.E.C., Martin, K.L., Morris, A.D., Shuttle worth, A.D., Rubdige, T., Anderson, D.W., Kelly, D.C. (1989) Molecular cloning and nucleotide sequence of the alpha toxin of Clostridium perfringens . Infect. Immun. 57:367-376.
    16. Uemura, T., Maekawa, T. and Sakaguchi, G. (1984). Biological Assay for C. perfringens enterotoxin with vero cells. Jpn. J. Vet. Sci., 46:715-720. 
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