Evaluation of three dilutions of E. coli antigen in immunogenicity in balb/c mice

Document Type : Full Research Paper

Authors

1 Department of Microbiology, Razi Vaccine and Serum Research Institute, Shiraz Branch, Agricultural Research, Education and Extension Organization (AREEO), Shiraz, Iran.

2 Department of Biology, Facoulty of Sciences, Shiraz Branch, Islamic Azad University, Shiraz, Iran.

Abstract

Enterotoxigenic Escherichia coli (E. coli) (ETEC) is the common cause of bacterial diarrhea and mortality in calves. Due to antibiotic resistant, study on vaccination as the main prevention method is important. In current study, three dilutions of E. coli K99 antigen were evaluated for immunization of balb/C mice. Bacterial culture was inactivated by formaldehyde and washed twice by sterile PBS. Sedimentation products were adjusted in three dilutions of 3 × 107, 3× 1011 and 3× 1013 CUF/m1 due to McFarland standard tubes. Three groups each consisting of 10 mice were set as experimental groups which received two subcutaneous injections doses of different concentrations with two weeks intervals. The control groups (10) received placebo. Mice blood samples were collected during 10 weeks and antibody titer was evaluated by Elisa. Serum titers in all three experimental groups increased from 4th to 8th week and then decreased gradually. No differences were observed between groups two and three, but differences were observed between groups one and two, and one and three. Immunization by whole cell antigen was not specific but was complete. On the other hand; immunization was carried out against each antigenic epitope. When the immune cells sites are occupied by antigenic epitopes and then saturated, excessive concentration of antigen does not have much impact on the immunogenicity.

Keywords

Main Subjects

References


1. Aguilar, J.C., Rodriguez, E,G. (2007). Vaccine adjuvants revisited. Vaccine. 25 (19): 3752- 3762.
2. Andrews, A. H., Blowey, R. W., Boyd, H. Eddy, R. G. (2004). Bovine medicine disease and husbandry of cattle. 2nd edition. Blackwell Publishing. AAAP.Pp: 186-230.
3. Badiei, K.h., Pourjafar, M., Ghane, M. (2011). Fecal E. coli F5 (k99) Antigen in Diarrheric calves of high and Average producing Holstein dairy cows. Global Vet. 5 (3): 175-1179.
4. Bertin, A. (2000). F41 antigen as a virulence factor in the infant mouse model of Escherichia coli diarrhoea. J. Gen. Microbiol. 131 (2): 3037-3045.
5. Crouch, F., Oliver., S., Francis, M.J. (2001). Serological, colostral and milk responses of cows vaccinated with a single dose of a combined vaccine against rotavirus, corono virus and Escherichia coli K99.Vet. Rec.149 (4): 105-108.
6. Duchet-Suchaux, M. (2002). Protective antigens against enterotoxigenic Escherichia coli 0101:K99, F41 in the infant mouse diarrhea model. Infect. Immun. 56 (6):1364-1370.
7. Duchet-Suchaux, M., Bertin, A., Dubray, G. (2006). Morphological description of surface structures on strain B41 of bovine enterotoxigenic Escherichia coli bearing both K99 and F41antigens. J. Gen. Microbiol. 134(1): 983-995.
8. Figueiredo, H.C.P., Large, A.P., Pereiaj Unior, F.N., Leite, R.C. (2004). Passive immunity in cattle againts ETEC. Serologic evaluation of a bacterin containing k99 and F41 Fimbria in colostrum of vaccinated Females and calf serum. Arq. Bras. Med. Vet. Zootec. 56 (12): 425-432.
9. Forman, A.J., Acres, S.d., Kapitany, R.A., Buckley, M.L., Stainer, D.W., Maxwell, M.A.B. (1981). The Immunogenicity of K99 Antigen in Whole cell Bacterins of Escherichia coli. Can. J. Comp. Med. 46 (4): 426-429.
10. Franck, S. M., Bosworth, B. T., Moon, H. W. (1998). Multiplex PCR for enterotoxigenic, attaching and effacing, and Shiga toxin-producing Escherichia coli strains from calves. J Clin. Microbiol. 36 (2): 1795-1797.
11. Germine , S.S., Ebied, M.H., Ibrahim, F.K., Mettias, K.N., Daoud, A.M. (2011). Field evalution of egg yolk antibodies in prevention and treatment of enteric coli bacilosis in calves. Benha Vet. Med. J., Special Issue 1: 108-114.
12. George, M., Garrity, E.D. (2005). The Gammaproteobacteria. Bergey's Manual of Systematic Bacteriology. 2B (2nd ed.). New York: Springer. British Library no. GBA561951.Pp: 1108-2.
13. Gorbach, S.L., Banwell, J.G., Chatterjee, B.D., Jacobs, B., Sack R.B. (1971). Acute undifferentiated human diarrhea in the tropics. I. Alterations in intestinal micrflora. J Clin Investig. 50 (3):881-9.
14. Johnson, A.M., Kaushik, R.S., Francis, D.H., Fleckenstein, J.M., Hardwidge, P.R. (2009). Heat-labile enterotoxin promotes Escherichia coli adherence to intestinal epithelial cells. J Bacteriol. 191(1):178-186.
15. Marion, D. (1988). Prodective against Enterotoxigenic Escherichia coli O101;k99,f41 in the infant mouse diarrhea model. Infect. Immun. 5(3): 1364-1370.
16. Morris, J. A., Thorns, C., Scott, A. C., Sojka, W. J., Wells, G. A. (2003). Adhesion in vitro and in vivo associated with an adhesive antigen (F41) produced by a K99 mutant of the reference strain Escherichia coli B41. Infect. Immun. 36(1):1146-1153.
17. Nagy, B., Fekete, P.Z. (2005). Enterotoxigenic Escherichia coli in veterinary medicine. Int J Med Microbiol. 295 (6): 443-454.
18. Ok, M., Guler, L., Turgut, K., Ok, U., Sen, I., Gunduz, K., Birdane, M.F.,Guzelbektes, H. (2008). The studies on the Aetiology of Diarrhoea in neonatal calves and determination of virulence gene markers of Escherichia coli Strains by multiplex PCR. Zoonoses Public Health. 56 (2): 94-101.
19. Olsvik, O., Strockbine, N. A. (1993). PCR detection of heat-stable, heat-labile, and Shiga-like toxin genes in Escherichia coli. In Diagnostic Molecular Microbiology. Principles and Appl. 233(3): 445-451.
20. Qadri, F., Svennerholm, A.M., Faruque, A.S.G., Bradley, S. R. (2005). Enterotoxigenic Escherichia coli in Developing Countries: Epidemiology, Microbiology, Clinical Features, Treatment, and Prevention. Clin. Microbiol. Rev. 18(5):465-83.
21. Rabbani, M., Khabardezfooli, M., Zahraee Salehi, T., Yoosefiramandi, A., Baahonar, A., Rezazadeh, F. (2007). Evaluation of antibodies against E. coli, rotavirus and coronavirus in serum of calves under one month of age with diarrhea and healthy calves. Vet Res J. 3 (2): 145-149.
22. Radostits, O.M., Gay, C.C., Blood, D.C. (2000). Veterinary Medicine, 9th Ed. W.B. Saunders; London. Pp: 734-738.
23. Shams, Z., Tahamtan, Y. , Pourbakhsh, A. , Hosseiny, M. H. , Kargar, M. Hayati, M. (2012) Detection of enterotoxigenic K99 (F5) and F41 from fecal sample of calves by molecular and serological methods. Comp. Clin. Pathol. 21(4): 475–478.
24. Walker, R.I., Steele, D., Aguado, T. (2007). Analysis of strategies to successfully vaccinate infants in developing countries against enterotoxigenic E. coli (ETEC) disease. Vaccine. 25 (3): 2545-2566.
25. Zhffa, A., Sslajka, E., Zajak, j. (1991). Antibody response in mice,rabbits and piges in response to vaccination with an inactivated oil vaccine coataining rotavirus and Escherichia coli strains with k88, k99 and 987p fimberial antigens. Vet. Med. 36(1): 423-431.
Veterinary Research & Biological Products
Volume 30, Issue 4
December 2017
Pages 40-49

Files

History

  • Receive Date: 20 August 2016
  • Revise Date: 31 January 2017
  • Accept Date: 01 February 2017

Share

How to cite

Statistics