Four cows were fed once a day either a Cocksfoot hay diet (H) or a diet consisting of 65% hay and 35% pelleted ground barley (HB). 15(NH4)2SO4 was continuously infused into the rumen as a microbial marker and ruminal digesta samples were collected during the 24-h postprandial period for the isolation of liquid-associated protozoa and bacteria (LAP, LAB) and particle-associated bacteria (PAB). There were marked differences between ruminal pH diurnal variations with diets H and HB. Irrespective of the diet and sampling time, the chemical composition (OM, N, DAPA, 15N) of the protozoa was clearly different from that of the bacteria (P < .001). The LAP contained more OM but less N and 15N than the bacterial fractions. The DAPA used to validate the isolation technique for the mixed ciliate population was not detected in protozoal fractions. The OM content of LAB was lower than that of PAB, whereas the N, DAPA, and 15N contents were higher. The observed effects of diet (P < .01) on LAP mean N contents were due to the different N contents of the LAP samples isolated 23 h after feeding and were correlated with the variation in the number of Endodiniomorphid protozoa (r = .72; P < .05). The N content of LAB was not affected (P > .05) by diet but that of the PAB was increased on diet HB (P < .05). The diet did not affect the 15N content of any of the three microbial populations. However, the 15N content of the bacteria decreased shortly after feeding (P < .001).

Summary ― Four naturally born lambs were placed in sterile isolators 24 h after birth, before the natural establishment of the cellulolytic microorganisms. At the age of 4 weeks, a cellulolytic bacterial population of approximately 10 a cells g-1 of rumen contents was established by inoculation with a 10-6 dilution of ruminal contents taken from an adult sheep. A pure culture of Neocallimastix frontalis MCH 3 and Piromyces communis FL was inoculated into the rumen 5 months after birth and a stable population of 10 3 -10 4 zoospores g-1 developed; the cellulolytic bacteria and fungi established populations in the 4 lambs that were similar to those observed in conventional animals. The presence of fungi led to an increase in the activity of most of the glycoside and polysaccharide hydrolases of the particle-associated microbial populations. However, this effect was not acompanied by an increase in the in sacco degradation of wheat straw or an increase in the volatile fatty acid concentration in the rumen contents.