The transmission of Salmonella Enteritidis (SE) in poultry is most often by the fecal-oral route, which can be attributed to the population density. Consequently, the pathogen triggers stress response and virulence factors deploying it to survive in hosts. Therefore, this study proposed to evaluate HPMCAS-coated microparticles containing the Ctx(Ile21)-Ha antimicrobial peptide against SE in laying hens chicks infection model to determine whether Ctx(Ile21)-Ha-utilization confers a benefit in the intestinal lumen, as well as whether limits systemic infection. Importantly, while assessing whether AMP utilization confers reduction of SE in liver, it was noted that there was statistical significance between groups A (control, no Ctx(Ile21)-Ha peptide) and B (2.5 mg of Ctx(Ile21)-Ha/kg) at 2 dpi, potentially indicating the Ctx(Ile21)-Ha effectiveness in the first stage of infection by SE. Remarkably, it was also detected a statistical significance (p -value <0.0001) with lower counts of SE (~ 0 CFU) in livers at 5, 7, and 14 dpi, regardless of Ctx(Ile21)-Ha dosage (2.5 mg or 5 mg/kg - group C). By using Chi-square test, the AMP effect on SE fecal excretion was evaluated. In this regard, it was noticed statistical significance (p < 0.05) among groups B and C in comparison with control group A, since those groups had lower bacterial excretion along 21 days. In summary, the role of HPMCAS-Ctx(Ile21)-Ha peptide microcapsules against S. Enteritidis in laying hen chicks infection model was unraveled, providing a satisfactory results against this pathogen.