This study was undertaken to evaluate and compare the efficacy of different multi-antigen vaccines, including heat-inactivated, whole lysate, and subunit (outer membrane proteins [OMPs]) C. jejuni vaccines along with the immunostimulant CpG ODN in controlling Campylobacter colonization in chickens. In the first trial, 125 μg of C. jejuni OMPs and 50 μg of CpG ODN were administered individually or in combination, either in ovo to chick embryos or subcutaneously (SC) to one-day-old chicks. In the second trial, different concentrations of C. jejuni antigens (heat-killed, whole lysate, and OMPs) were administered SC to one-day-old chicks. The results of the first trial revealed that SC immunization with the combination of CpG ODN and C. jejuni OMPs elevated interferon (IFN)-γ, interleukin (IL)-1β, and IL-13 gene expression in the spleen, significantly increased serum IgM and IgY antibody levels, and reduced cecal C. jejuni counts by approximately 1.2 log₁₀. In contrast, in ovo immunization did not elicit immune responses or confer protection against Campylobacter. The results of the second trial showed that SC immunization with C. jejuni whole lysate or 200 μg OMPs reduced C. jejuni counts by approximately 1.4 and 1.1 log₁₀, respectively. In conclusion, C. jejuni lysate and OMPs are promising vaccine antigens for reducing Campylobacter colonization in chickens.

Research on the immunological development of lymphoid organs in chicks has been extensive, yet a significant gap exists in our understanding of innate immunity during embryonic life within the intestinal tract. This study investigated the developmental trajectory of intestinal immunity in chick embryos by evaluating basal gene expression levels of key immune markers at embryonic days (ED) 14, 17, and 20. The results indicated variable expression levels of cytokines, antimicrobial peptides (AMPs), and Toll-like receptor (TLRs) genes throughout the intestinal tract. Most cytokines and chemokines exhibited elevated expression in the cecum, while AMPs, including avian-β-defensins (AvBDs) and cathelicidins (CATHs) genes, showed increased levels in the jejunum at ED20. The findings from the developmental trajectory analysis of these genes revealed elevated expression levels of cytokines, including interferon (IFN)-γ, interleukin (IL)-6, IL-13, and transforming-growth factor (TGF)-β in the cecum at ED20. However, no consistent patterns were observed for AvBDs, CATHs, and TLRs, as their expression varied across different developmental stages of the chick embryo. These findings significantly contribute to our understanding of intestinal immune system development in chick embryos and provide a foundation for further research aimed at enhancing immune capabilities, especially in segments with lower expression levels of immunomodulatory genes.