Sera of young sows from 135 closed Belgian pig breeding farms were examined for the presence of serum antibodies specific for the F4 fimbrial antigen of enterotoxigenic Escherichia coli (ETEC). Since 80% of all pig farms in Belgium are located in the provinces West-Vlaanderen (44%, approximately 18 farms/10 km2), Oost-Vlaanderen (20%, approximately 9 farms/10 km2), Antwerpen (10%, approximately 5 farms/10 km2) and Vlaams-Brabant (6%, approximately 3 farms/10 km2), the farms examined were randomly selected in these four regions. On 68% of all tested farms, sows were not vaccinated against enteric colibacillosis. In general, 65% of these non-vaccinated farms were F4-seropositive (mean optical density [OD405] >0.5) of which 38% were weakly (mean OD405 = 0.5-1.0), 21% moderately (mean OD405 = 1.0-2.0) and 6% strongly positive (mean OD405 > or =2.0). These percentages showed major differences between the four provinces. In Vlaams-Brabant, only 31% of non-vaccinated farms were seropositive, of which most were regarded as weakly positive. In Antwerpen, Oost- and West-Vlaanderen however, this percentage was clearly higher and respectively 68, 71, and 79% of non-vaccinated farms were seropositive, of which most were weakly positive. These observations indicate that F4+ ETEC is widely spread and highly prevalent on non-vaccinated pig breeding farms in Belgium. Moreover, an increasing F4-seropositivity with increasing pig farm density suggests a possible influence of the swine density on the prevalence of F4+ ETEC infections.