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Egg yolk antibodies, called IgY, were isolated from the egg yolks of chickens immunized with enterotoxigenic Escherichia coli (ETEC) 987P. Specific binding activity of IgY against ETEC 987P was found. Whole cell titers remained relatively high during the immunization period (up to 19 weeks). The IgY antibodies cross-reacted with other members of Enterobacteriaceae, Escherichia coli O157:H7, Salmonella enteritidis, and S. typhimurium by 28.3%, 21.1% and 19.7%, respectively. Specific IgY with a concentration higher than 0.54 mg/ml was found to inhibit the growth of ETEC 987P in a liquid medium. The difference in bacterial growth between the specific and the non-specific IgY was 6.1 log CFU/ml after 8 hr incubation. The specific binding activity of IgY to bacteria was further evaluated by immunofluorescence and immunoelectron microscopy, showing the binding of specific IgY to the bacterial surface. The immunoelectron microscopic observation revealed structural alterations on the bacterial surface bound by specific IgY. This proves that specific binding activity of IgY against ETEC 987P whole cells resulted in growth inhibition of effect ETEC 987P in vitro.