bThis study determined the effect of feed fermentation with Lactobacillus reuteri on growth performance and the abundance of enterotoxigenic Escherichia coli (ETEC) in weanling piglets. L. reuteri strains produce reuteran or levan, exopolysaccharides that inhibit ETEC adhesion to the mucosa, and feed fermentation was conducted under conditions supporting exopolysaccharide formation and under conditions not supporting exopolysaccharide formation. Diets were chosen to assess the impact of organic acids and the impact of viable L. reuteri bacteria. Fecal samples were taken throughout 3 weeks of feeding; at the end of the 21-day feeding period, animals were euthanized to sample the gut digesta. The feed intake was reduced in pigs fed diets containing exopolysaccharides; however, feed efficiencies did not differ among the diets. Quantification of L. reuteri by quantitative PCR (qPCR) detected the two strains used for feed fermentation throughout the intestinal tract. Quantification of E. coli and ETEC virulence factors by qPCR demonstrated that fermented diets containing reuteran significantly (P < 0.05) reduced the copy numbers of genes for E. coli and the heat-stable enterotoxin in feces compared to those achieved with the control diet. Any fermented feed significantly (P < 0.05) reduced the abundance of E. coli and the heat-stable enterotoxin in colonic digesta at 21 days; reuteran-containing diets reduced the copy numbers of the genes for E. coli and the heat-stable enterotoxin below the detection limit in samples from the ileum, the cecum, and the colon. In conclusion, feed fermentation with L. reuteri reduced the level of colonization of weaning piglets with ETEC, and feed fermentation supplied concentrations of reuteran that may specifically contribute to the effect on ETEC.T he combined stress of weaning and movement to a different environment increases the potential for poor growth performance, nutrient malabsorption, and diseases in piglets (1-3). Diarrhea caused by enterotoxigenic Escherichia coli (ETEC) is a major disease of weaning piglets (4). ETEC establishes infection by specific fimbria mediating intestinal attachment and colonization (4-7). Following colonization, ETEC produces enterotoxins that induce watery diarrhea (8, 9). Control of ETEC infections of weanling piglets is currently achieved by antibiotics (10). The use of antibiotics in animal production, however, selects for antibiotic-resistant intestinal microbiota and favors the transfer of antibiotic resistance genes from livestock microbiota to human pathogens (7). Consequently, antimicrobial growth promoters have been banned in several jurisdictions (11), challenging the livestock industry to replace feed antibiotics without compromising animal performance or animal health.Probiotic bacteria, prebiotics, organic acids, or antiadhesive glycans were proposed to replace feed antibiotics in pig production for improved control of ETEC (4, 6). Organic acids were shown to reduce postweaning diarrhea in pigs (12). Probiotics also decreased the inc...