The objective of this study was to investigate the effect of short-term elevated ambient temperature on ruminal volatile fatty acid (VFA) dynamics and rumen epithelium gene expression associated with the transport and metabolism of VFA. Eight ruminally cannulated Holstein heifers (200 kg) were used in a factorial, repeated measures experiment with two treatments and two periods. During the first period, animals were provided with feed ad libitum and housed at 20 degrees C. During the second period, one group (HS) was housed at 30 degrees C and fed ad libitum. The other group (PF) was housed at 20 degrees C and pair-fed to match the intake of the HS group. During each period, animals were kept on treatment for 10 day, with sample collection on the final day. In the second period, indicators of heat stress were significantly different between PF and HS animals (P<0.05). There was a thermal environment effect on butyrate production (P<0.01) that was not associated with feed intake (P=0.43). Butyrate absorption decreased in HS animals (P<0.05) but increased in PF animals (P<0.05) from period 1 to period 2. There was a feed intake effect on BHD1 expression (P=0.04) and a tendency for a thermal environment effect (P=0.08), with expression increasing in both cases. Expression of MCT4 was affected by feed intake (P=0.003) as were all NHE genes (NHE1, NHE2, and NHE3; P<0.05). These results indicate that with low feed intake and heat stress, there are shifts in rumen VFA dynamics and in the capacity of the rumen epithelium to absorb and transport VFA.