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The aim of this study was to evaluate the ability of unfractionated hen egg yolk to prevent the attachment of Salmonella typhimurium to murine intestinal epithelial cells in vitro. Inhibition of adherence was examined microscopically and by an ELISA. Both methods showed that egg yolk from immunized and unimmunized hens significantly reduced bacterial adherence. Optical density (OD) readings Ϯ Ϯ Ϯ Ϯ Ϯ SD of 0.825 Ϯ Ϯ Ϯ Ϯ Ϯ 0.016 for untreated bacteria were reduced to 0.335 Ϯ Ϯ Ϯ Ϯ Ϯ 0.024 and 0.267 Ϯ Ϯ Ϯ Ϯ Ϯ 0.021 for bacteria pretreated with egg yolk from immunized hens and unimmunized hens, respectively. Microscopic analysis showed that egg yolk from either immunized or unimmunized hens reduced bacterial adherence from 17Ϯ Ϯ Ϯ Ϯ Ϯ 2.7 bacteria per epithelial cell to less than 4 bacteria per epithelial cell. These results show that hen egg yolk significantly inhibits adherence of S. typhimurium to intestinal epithelial cells in vitro. Boosting of specific antibody levels does not enhance this antiadhesive effect, suggesting that egg yolk contains novel antiadhesive factors.