One induced and one spontaneous serum-resistant mutant were derived from smooth serum-sensitive Escherichia coli strains. There was little difference in the yield or the 0-side-chain-sugar to core-sugar ratio of lipopolysaccharides from the mutants compared with the parental strains. The mutations to serum resistance were not accompanied by increases in either the amount or haemagglutinationinhibiting activity of acidic polysaccharides extracted from the strains.Two colonially rough forms, designated 17fa and 17gb, were isolated from an aged culture of serum-resistant mutant 17. Escherichia coli 17fa appeared to be a som mutant and was rapidly killed by serum. Escherichia coli 17gb retained serological 0-specificity, and 17gb lipopolysaccharide contained a full complement of 0-side-chains; the strain was killed by serum but only after a delay of I h.Escherichia coli K-negative 0 8 donor Hfryj, which was serum-resistant, was crossed with rough serum-sensitive E. coli strain ~4 7 0 and his+ recombinants were selected. The majority of his+ recombinants inherited a full complement of lipopolysaccharide 0-side-chains but were killed by serum after a delay of I to 2 h.These results suggest that lipopolysaccharide 0-side-chains are responsible for a delay in the killing by normal human serum of smooth E. coli strains but that other factors determine full serum resistance. No evidence was found of a role for K-antigens in serum resistance.