Comparison of various amino acids for estimation of microbial nitrogen in digesta.

ABSTRACT

Methods of estimating microbial N in digesta were compared, and two simplified methods were described for separation of diaminopimelic acid (DAP) from methionine in acid hydrolysates of biological samples using an automated amino acid analyzer. Recoveries of DAP when added to bacterial, abomasal and feed samples were 92 to 98%. All samples analyzed (bacteria, protozoa and several feedstuffs) contained detectable DAP (or another amino acid with similar elution time). Apparent DAP-N (percentage of total sample N) in feeds ranged from 12 to 78% of bacterial DAP-N concentration. These values are greater than can be accounted for by bacterial contamination. Correction for apparent dietary DAP was appropriate when using this microbial marker. Reasonable estimation of bacterial N was made in abomasal digesta of steers fed high-grain diets using DAP, lysine or combinations of DAP and leucine or lysine and leucine. There was excellent agreement among these marker methods in ranking dietary treatment regimens with respect to bacterial N present in the abomasum. The use of leucine alone, or the combination of DAP and lysine, gave nonrealistic values. Two-amino-ethylphosphonic acid (AEP) was not detected in acid hydrolysates of mixed ruminal protozoa or abomasal digesta. Two other ninhydrin-reactive compounds with elution times close to AEP were detected in all samples analyzed, which may have been identified as AEP in other studies. It was concluded that corrected DAP and lysine were the best estimates of bacterial N and AEP was precluded as a marker of protozoal N.