This study was conducted to determine the effect of formaldehyde treatment of barley on the rate of microbial starch digestion. Ground barley was left untreated (UB) or treated with 0.5% formaldehyde (TB) and incubated in vials with an inoculum consisting of a homogenate of rumen contents and Bryant's anaerobic media (1:1). Separate vials were analyzed for starch, ammonia, pH and volatile fatty acids (VFA) after 0, 4, 8, 12, 18, 24, 36 and 48 h. The digestion of starch was lower (P < 0.05) in TB than in UB between 4 and 24 h. Accumulation of NH3 was lower and pH was higher in TB vials (P < 0.05) than in UB vials. VFA production and fermentation efficiency were lower (P < 0.05) in TB than in UB. Formaldehyde treatment of barley reduced (P < 0.05) the production of propionate and enhanced (P < 0.05) the production of butyrate in comparison with UB. Examination by scanning electron microscopy (SEM) showed that the initial colonization of TB was inhibited in comparison with that of UB. The distinct shape of the endosperm cells of TB (12 h) were readily recognizable, but the endosperm cells of UB (12 h) were not readily distinguishable. SEM of TB (24 h) showed that the protein matrix of the endosperm cell is resistant to microbial digestion. The formaldehyde-induced reduction in starch digestion appears to result from the protein matrix inhibiting the access of bacteria to underlying starch granules. Key words: Starch, rumen bacteria, formaldehyde, barley