Mycobacterium avium subspecies paratuberculosis (MAP), the causative agent of Johne’s disease and a potential contributor to Crohn’s disease, presents a significant challenge due to its resistance to conventional antibiotics. This necessitates the development of innovative strategies for prevention and treatment. This study aimed to evaluate the antibacterial activity of pathogen-specific antibodies derived from chicken egg yolks (IgY) and the postbiotics from lactic acid bacteria against MAP. Immunoglobulin Y antibodies were produced by immunizing hens with formalin-killed MAP strain antigens. The IgY was extracted, purified, and the anti-MAP titers were quantified by indirect ELISA. The Minimum Inhibitory Concentration (MIC) of different concentrations of specific anti-MAP IgY and the mixture of postbiotics (from four different probiotic strains Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus acidophilus, and Pediococcus acidilactici) individually and in combination against MAP was determined at various time intervals. Anti-MAP IgY titers in egg yolks increased within two weeks of immunization, reaching peak levels at six weeks. Growth inhibition assays revealed that postbiotics concentration as low as 6.25 mg mL-1 effectively inhibited MAP growth. Anti-MAP IgY demonstrated antibacterial activity with a MIC of 50 mg mL-1, while the combined IgY-posbiotics treatment achieved MAP growth inhibition at a MIC of 3.125 mg mL-1. The findings of the study suggests that combination therapy with specific IgY and postbiotics may be a promising preventive strategy for controlling MAP infections. Further in vivo studies are needed to elucidate the underlying mechanisms and optimize the application of this approach for broader use in veterinary and human medicine.