The optimization of a prevention program utilizing a novel trivalent inactivated Salmonella bacterial vaccine to protect poultry from Salmonella infections was evaluated in this study. A total of 50 brown nick layers were divided into 5 groups, A to E, each containing 10 hens. Group B hens were immunized with the SG9R vaccine, group C chickens were immunized with the trivalent inactivated Salmonella bacterial vaccine, and group D hens were primed with SG9R and boosted with the trivalent inactivated Salmonella bacterial vaccine. Group E hens were injected with sterile PBS. All hens in groups B to E were orally challenged with a mixture of wild-type S. Enteritidis, S. Typhimurium, and S. Gallinarum (approximately 6×10⁹ CFU/ 0.2 ml/bird). Serum IgG titers, CD3+CD4+ T-cell levels, and CD3+CD8+ T-cell levels in group D were significantly higher than those in group A. Additionally, all animals in groups A to D showed no clinical symptoms and survived after the virulent challenges, whereas all chickens in group E died following the challenge. The challenge strains of S. Enteritidis, S. Typhimurium, and S. Gallinarum were not isolated from the liver, spleen, cecum, and cloaca of group D chickens. These findings indicate that priming with SG9R and boosting with a trivalent inactivated Salmonella bacterial vaccine can be an effective approach for preventing Salmonella infections by inducing robust protective humoral and cellular immune responses in chickens.

Transplantation is considered to be a very useful approach to improve human welfare and to prolong life-span. Heterologous organ transplantation using pig organs which are similar to human beings and easy to make mass-production has known as one of the alternatives. To ensure potential usage of the pig organ for transplantation application, it is essentially required to generate transgenic pig modifying immuno-related genes. Previously, we reported production of heterozygous α 1,3-galactosyltransferase (GalT) knock-out and human membrane cofactor protein (MCP) expressing pig (GalT-MCP/+), which is enforced for suppression of hyperacute and acute immunological rejection. In this study, we reported generation of homozygous pig (GalT-MCP/-MCP) by crossbreeding GalT-MCP/+ pigs. Two female founders gave birth to six of GalT-MCP/-MCP, and seven GalT-MCP/+ pigs. We performed quantitative real-time PCR, western blot, and flow cytometry analyses to confirm GalT and MCP expression. We showed that fibroblasts of the GalT-MCP/-MCP pig do not express GalT and its product Gal antigen, while efficiently express MCP. We also showed no expression of GalT, otherwise expression of MCP at heart, kidney, liver and pancreas of transgenic pig. Taken together, we suggest that the GalT-MCP/-MCP pig is a useful candidate to apply xenotransplantation study.