Four multiple-cannulated steers (340kg) were used in a 4×4 Latin square design with a 2×2 factorial arrangement of treatments. Steers were fed a diet of 50% ground hay and 50% concentrate at two intakes (1.4 and 2.1% of BW), with urea and 15N-enriched ammonium sulfate infused continuously into the rumen at .4 or 1.2% of diet DM. Ratios of purines and diaminopimelic acid-N to N in fluid-associated and particulate-associated bacteria and in protozoa were similar among treatments but were lower for protozoa than for bacteria. Diaminopimelic acid-N:N was higher for fluid-associated vs. particulate-associated bacteria. Enrichment of 15N was similar between bacteria among treatments and was 30% lower for protozoa. Turnover rates of 15N in bacteria, NH3N, and non-NH3N pools were faster for steers infused with 1.2 than those infused with .4% urea, indicating less efficient usage of ammonia with higher urea. A method is described to estimate the proportion of duodenal nitrogen comprising bacterial and protozoal nitrogen.