Protection Against Salmonella Typhimurium, Salmonella Gallinarum, and Salmonella Enteritidis Infection in Layer Chickens Conferred by a Live Attenuated Salmonella Typhimurium Strain

  • Lee, John Hwa (College of Veterinary Medicine and Bio-Safety Research Institute, Chonbuk National University)
  • Received : 2014.11.10
  • Accepted : 2014.12.29
  • Published : 2015.02.28


In the present study, we investigated the protection conferred by a live attenuated Salmonella enterica serovar Typhimurium (ST) strain against Salmonella Typhimurium, Salmonella Gallinarum (SG), and Salmonella Enteritidis (SE) infection in layer chickens. Birds were orally primed with the attenuated ST strain at 7 days of age and then boosted at 4 weeks post prime immunization (PPI). Sequential monitoring of plasma IgG and mucosal secretory IgA (sIgA) levels revealed that inoculation with ST induced a significant antibody response to antigens against ST, SE, and SG. Moreover, significant lymphoproliferative responses to the 3 Salmonella serovars were observed in the immunized group. We also investigated protection against virulent ST, SE, and SG strain challenge. Upon virulent SG challenge, the immunized group showed significantly reduced mortality compared to the non-immunized group. The reduced persistence of the virulent ST and SE challenge strains in the liver, spleen, and cecal tissues of the immunized group suggests that immunization with the attenuated ST strain may not only protect against ST infection but can also confer cross protection against SE and SG infection.


Supported by : National Research Foundation (NRF)


  1. Silva, E. N., and A. Duarte. 2002. Salmonella Enteritidis in poultry: retrospective in Brazil. Braz. J. Poult. Sci. 4: 85-100.
  2. Forshell, L. P., and M. Wierup. 2006. Salmonella contamination: a significant challenge to the global marketing of animal food products. Rev. Sci. Tech. 25: 541-554.
  3. Duguid, J. P., and R. A. North. 1991. Eggs and Salmonella food-poisoning: an evaluation. J. Med. Microbiol. 34: 65-72.
  4. Rodrigue, D. C., R. V. Tauxe, and B. Rowe. 1990. International increase in Salmonella enteritidis: a new pandemic? Epidemiol. Infect. 105: 21-27.
  5. Poppe C. 2000. Salmonella Infections in the Domestic Fowl In: Wray C, Wray A, editors. Salmonella in domestic animals, Wallingford: CABI Publishing; p. 107-132.
  6. Carvajal, B. G., U. Methner, J. Pieper, and A. Berndt. 2008. Effects of Salmonella enterica serovar Enteritidis on cellular recruitment and cytokine gene expression in caecum of vaccinated chickens. Vaccine 26: 5423-5433.
  7. Chappell, L., P. Kaiser, P. Barrow, M. A. Jones, C. Johnston, and P. Wigley. 2009. The immunobiology of avian systemic salmonellosis. Vet. Immunol. Immunopathol. 128: 53-59.
  8. Velge, P., A. Cloeckaert, and P. Barrow. 2005. Emergence of Salmonella epidemics: the problems related to Salmonella enterica serotype Enteritidis and multiple antibiotic resistance in other major serotypes. Vet. Res. 36: 267-288.
  9. McGruder, E. D., P. M. Ray, G. I. Tellez, M. H. Kogut, D. E. Corrier, J. R. DeLoach, and B. M. Hargis. 1993. Salmonella enteritidis immune leukocyte-stimulated soluble factors: effects on increased resistance to Salmonella organ invasion in day-old Leghorn chicks. Poult. Sci. 72: 2264-2271.
  10. Wigley, P., S. Hulme, C. Powers, R. Beal, A. Smith, and P. Barrow. 2005. Oral infection with the Salmonella enterica serovar Gallinarum 9R attenuated live vaccine as a model to characterise immunity to fowl typhoid in the chicken. BMC. Vet. Res. 1: 2.
  11. Girard, M. P., D. Steele, C. L. Chaignat, and M. P. Kieny. 2006. A review of vaccine research and development: human enteric infections. Vaccine 24: 2732-2750.
  12. Sieve, A. N., K. D. Meeks, S. Bodhankar, S. Lee, J. K. Kolls, J. W. Simecka, and R. E. Berg. 2009. A novel IL-17-dependent mechanism of cross protection: respiratory infection with mycoplasma protects against a secondary listeria infection. Eur. J. Immunol. 39: 426-438.
  13. Calenge, F., P. Kaiser, A. Vignal, and C. Beaumont. 2010. Genetic control of resistance to salmonellosis and to Salmonella carrier-state in fowl: a review. Genet. Sel. Evol. 42: 11.
  14. Gantois, I., R. Ducatelle, L. Timbermont, F. Boyen, L. Bohez, F. Haesebrouck, F. Pasmans, and I. F. van. 2006. Oral immunisation of laying hens with the live vaccine strains of TAD Salmonella vac E and TAD Salmonella vac T reduces internal egg contamination with Salmonella Enteritidis. Vaccine 24: 6250-6255.
  15. Pathangey, L., J. J. Kohler, R. Isoda, and T. A. Brown. 2009. Effect of expression level on immune responses to recombinant oral Salmonella enterica serovar Typhimurium vaccines. Vaccine 27: 2707-2711.
  16. Tan, S., C. L. Gyles, and B. N. Wilkie. 1997. Evaluation of an aroA mutant Salmonella typhimurium vaccine in chickens using modified semisolid Rappaport Vassiliadis medium to monitor faecal shedding. Vet. Microbiol. 54: 247-254.
  17. Tellez, G. I., M. H. Kogut, and B. M. Hargis. 1993. Immunoprophylaxis of Salmonella enteritidis infection by lymphokines in Leghorn chicks. Avian Dis. 37: 1062-1070.
  18. Chaudhari, A. A., S. W. Kim, K. Matsuda, and J. H. Lee. 2011. Safety evaluation and immunogenicity of arabinosebased conditional lethal Salmonella Gallinarum mutant unable to survive ex vivo as a vaccine candidate for protection against fowl typhoid. Avian Dis. 55: 165-171.
  19. Matsuda, K., A. A. Chaudhari, and J. H. Lee. 2011. Evaluation of safety and protection efficacy on cpxR and lon deleted mutant of Salmonella Gallinarum as a live vaccine candidate for fowl typhoid. Vaccine 29: 668-674.
  20. Nandre, R. M., K. Matsuda, A. A. Chaudhari, B. Kim, and J. H. Lee. 2012. A genetically engineered derivative of Salmonella Enteritidis as a novel live vaccine candidate for salmonellosis in chickens. Res. Vet. Sci. 93: 596-603.
  21. Matulova, M., H. Havlickova, F. Sisak, V. Babak, and I. Rychlik. 2013. SPI1 defective mutants of Salmonella enterica induce cross-protective immunity in chickens against challenge with serovars Typhimurium and Enteritidis. Vaccine 31: 3156-3162.
  22. Wahid, R., R. Simon, S. J. Zafar, M. M. Levine, and M. B. Sztein. 2012. Live oral typhoid vaccine Ty21a induces cross-reactive humoral immune responses against Salmonella enterica serovar Paratyphi A and S. Paratyphi B in humans. Clin. Vaccine Immunol. 19: 825-834.
  23. Mohler, V. L., D. M. Heithoff, M. J. Mahan, K. H. Walker, M. A. Hornitzky, L. W. Shum, K. J. Makin, and J. K. House. 2008. Cross-protective immunity conferred by a DNA adenine methylase deficient Salmonella enterica serovar Typhimurium vaccine in calves challenged with Salmonella serovar Newport. Vaccine 26: 1751-1758.
  24. Ansaldi, F., S. Bacilieri, P. Durando, L. Sticchi, L. Valle, E. Montomoli, G. Icardi, R. Gasparini, and P. Crovari. 2008. Cross-protection by MF59-adjuvanted influenza vaccine: neutralizing and haemagglutination-inhibiting antibody activity against A(H3N2) drifted influenza viruses. Vaccine 26: 1525-1529.
  25. Beal, R. K., P. Wigley, C. Powers, P. A. Barrow, and A. L. Smith. 2006. Cross-reactive cellular and humoral immune responses to Salmonella enterica serovars Typhimurium and Enteritidis are associated with protection to heterologous re-challenge. Vet. Immunol. Immunopathol. 114: 84-93.
  26. Carrat, F., and A. Flahault. 2007. Influenza vaccine: the challenge of antigenic drift. Vaccine 25: 6852-6862.
  27. Chacana, P. A., and H. R. Terzolo. 2006. Protection conferred by a live Salmonella Enteritidis vaccine against fowl typhoid in laying hens. Avian Dis. 50: 280-283.
  28. Roland, K., R. Curtiss, III, and D. Sizemore. 1999. Construction and evaluation of a delta cya delta crp Salmonella typhimurium strain expressing avian pathogenic Escherichia coli O78 LPS as a vaccine to prevent airsacculitis in chickens. Avian Dis. 43: 429-441.
  29. Chaudhari, A. A., and J. H. Lee. 2013. Evaluation of the adjuvant effect of Salmonella-based Escherichia coli heat-labile toxin B subunits on the efficacy of a live Salmonella-delivered avian pathogenic Escherichia coli vaccine. Avian Pathol. 42: 365-372.
  30. Chaudhari, A. A., K. Matsuda, and J. H. Lee. 2013. Construction of an attenuated Salmonella delivery system harboring genes encoding various virulence factors of avian pathogenic Escherichia coli and its potential as a candidate vaccine for chicken colibacillosis. Avian Dis. 57: 88-96.
  31. Dueger, E. L., J. K. House, D. M. Heithoff, and M. J. Mahan. 2001. Salmonella DNA adenine methylase mutants elicit protective immune responses to homologous and heterologous serovars in chickens. Infect. Immun. 69: 7950-7954.
  32. Barrow, P. A., M. A. Lovell, and A. Berchieri. 1990. Immunisation of laying hens against Salmonella enteritidis with live attenuated vaccines. Vet. Rec. 126: 241-242.
  33. Nassar, T. J., H. M. al-Nakhli, and Z. H. al-Ogaily. 1994. Use of live and inactivated Salmonella enteritidis phage type 4 vaccines to immunise laying hens against experimental infection. Rev. Sci. Tech. 13: 855-867.
  34. Kim, S. W., K. H. Moon, H. S. Baik, H. Y. Kang, S. K. Kim, J. D. Bahk, J. Hur, and J. H. Lee. 2009. Changes of physiological and biochemical properties of Salmonella enterica serovar Typhimurium by deletion of cpxR and lon genes using allelic exchange method. J. Microbiol. Methods 79: 314-320.
  35. Lindberg, A. A., T. Segall, A. Weintraub, and B. A. Stocker. 1993. Antibody response and protection against challenge in mice vaccinated intraperitoneally with a live aroA O4-O9 hybrid Salmonella dublin strain. Infect. Immun. 61: 1211-1221.
  36. Alvarez, J., M. Sota, A. B. Vivanco, I. Perales, R. Cisterna, A. Rementeria, and J. Garaizar. 2004. Development of a multiplex PCR technique for detection and epidemiological typing of Salmonella in human clinical samples. J. Clin. Microbiol. 42: 1734-1738.
  37. Betancor, L., F. Schelotto, M. Fernandez, M. Pereira, A. Rial, and J. A. Chabalgoity. 2005. An attenuated Salmonella Enteritidis strain derivative of the main genotype circulating in Uruguay is an effective vaccine for chickens. Vet. Microbiol. 107: 81-89.
  38. Porter, R. E., Jr. and P. S. Holt. 1992. Use of a pilocarpinebased lavage procedure to study secretory immunoglobulin concentration in the alimentary tract of White Leghorn chickens. Avian Dis. 36: 529-536.
  39. Gogal, R. M., Jr., S. A. Ahmed, and C. T. Larsen. 1997. Analysis of avian lymphocyte proliferation by a new, simple, nonradioactive assay (lympho-pro). Avian Dis. 41: 714-725.
  40. Centers for Disease Control and Prevention (CDC). 1996. Outbreak of Salmonella serotype Typhimurium associated with eating ground beef-- Wisconsin. JAMA 275: 353-354.
  41. Roels, T. H., P. A. Frazak, J. J. Kazmierczak, W. R. Mackenzie, M. E. Proctor, T. A. Kurzynski, and J. P. Davis. 1997. Incomplete sanitation of a meat grinder and ingestion of raw ground beef: contributing factors to a large outbreak of Salmonella Typhimurium infection. Epidemiol. Infect. 119: 127-134.
  42. Abd El Ghany M., A. Jansen, S. Clare, L. Hall, D. Pickard, R. A. Kingsley, and G. Dougan. 2007. Candidate live, attenuated Salmonella enterica serotype Typhimurium vaccines with reduced fecal shedding are immunogenic and effective oral vaccines. Infect. Immun. 75: 1835-1842.
  43. Kotton, C. N., and E. L. Hohmann. 2004. Enteric pathogens as vaccine vectors for foreign antigen delivery. Infect. Immun. 72: 5535-5547.
  44. Park, J. H., H. W. Sung, B. I. Yoon, and H. M. Kwon. 2009. Protection of chicken against very virulent IBDV provided by in ovo priming with DNA vaccine and boosting with killed vaccine and the adjuvant effects of plasmid-encoded chicken interleukin-2 and interferon-gamma. J. Vet. Sci. 10: 131-139.
  45. Rana, N., and R. C. Kulshreshtha. 2006. Cell-mediated and humoral immune responses to a virulent plasmid-cured mutant strain of Salmonella enterica serotype gallinarum in broiler chickens. Vet. Microbiol. 115: 156-162.