An immunogenic Salmonella ghost confers protection against internal organ colonization and egg contamination

The tightly regulated expression of the PhiX174 lysis gene E from a multi-copy plasmid led to the stable production of an Salmonella Enteritidis bacterial ghost. The present study was conducted to evaluate induction of the humoral and cell-mediated immune responses induced after single or double intramuscular immunization with the S. Enteritidis ghost and to assess its protective effect on colonization of the intestinal tract, visceral and reproductive organs, internal egg contamination, and egg production of laying chickens. A total of 60 chickens were equally divided into three groups (n = 20); group A (non-immunized control), group B (immunized at 8 and 16 weeks of age) and group C (immunized at 16th week of age). Chickens from both immunized groups B and C demonstrated significant increases in plasma IgG, intestinal secretory IgA levels, and antigen-specific lymphocyte proliferative responses. The population of CD3+CD4+ positive T cells in the immunized chickens was also significantly increased after immunization and virulent challenge. In addition, the immunized groups B and C showed significantly higher egg production and a lower percentage of S. Enteritidis contaminated eggs after challenge compared to those of group A. A comparison of challenge strain isolation from the immunized-challenged and non-immunized-challenged layer hens showed that the double immunization group induced excellent protection against intestinal, liver, splenic, and ovarian Salmonella colonization; however, the single immunized chickens showed lower counts only in the splenic and ovarian organs. Overall, the data give compelling evidence that vaccination with the S. Enteritidis ghost induced robust protective immunity against experimental avian salmonellosis and may contribute to the reduce incidence of egg contamination.