Use of radioactive nitrogen in studying nitrogen fixation in bacterial cells and their extracts.
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The present study compares estimates of rumen microbial-N production derived from duodenal flow measurements (15N and purine bases) with those from measurements of the urinary excretion of purine derivatives. Four Rasa Aragonesa ewes fitted with simple cannulas in the rumen and proximal duodenum were used. Four diets consisting of 550 g lucerne (Medicago sativa) hay/d as sole feed or supplemented with 220, 400 and 550 g rolled barley grain/d were given in a 4 x 4 random factorial arrangement. Duodenal digesta flows were determined by the dual-phase marker technique during continuous intraruminal infusions of Co-EDTA and Yb-acetate. Microbial contribution to the non-NH3 N (NAN) flow was estimated from 15N enrichment and purines: N ratio in duodenal digesta and bacterial fractions isolated from the rumen content. Whole tract organic matter (OM) digestibility and duodenal flow of OM and NAN increased (P < 0.001) with the level of barley supplementation. Digestible OM intake ranged from 19.0 to 42.7 g/kg metabolic weight (W0.75) and the duodenal flow of purine bases and the urinary excretion of allantoin increased linearly (P < 0.001) from minimum values of 7.47 (SD 1.524) and 4.65 (SD 0.705) mmol/d respectively on the basal diet to 18.20 (SD 1.751) and 11.62 (SD 0.214) mmol/d on the 400 g barley diet; a further increase in barley supplementation decreased both variables (13.50 (SD 2.334) and 8.77 (SD 0.617) mmol/d respectively). Urinary excretion of uric acid and hypoxanthine showed a slight but significant increase (P < 0.05) over all levels of barley. Molar recoveries of duodenal purine bases as purine derivatives or allantoin in the urine were 0.78 (SD 0.156) and 0.65 (SD 0.130) respectively. The increase on barley supplementation significantly augmented microbial-N, but large differences between microbial markers employed were observed. Mean values of microbial-N estimated from the duodenal purine bases or urinary allantoin excretion were on average 18 and 29% lower than those measured by 15N.
Broilers were fed 15N labelled wheat and the concentration of the labelled isotope in muscular tissue was determined. A negative correlation was found to exist between the percentage proportion of N in the muscle fractions and the level of 15N frequency established 12 hrs after administration of the labelled ration. Higher rates of tracer incorporation were observed in the muscles of legs and other muscles (predominatly red muscles) than in pectoral muscles (white muscles). The heart was shown to exhibit the lowest N content and the highest level of 15N frequency of all muscle fractions. In view of the fact that some specific difficulties will arise when certain factors (trace elements, vitamins, ergotropic substances, genetic factors) which influence the N metabolism of fattening poultry, are estimated on the basis of the conventional N balance trials the present tracer technique is recommended for use as an alternative method in studies of the kind just described. In the present trial 20% of the wheat N were utilized for the production of meat N in muscular tissue.
After a 45-hour infusion of 15N-lysine and 15N-glycine resp. with pigs of 25 kg live weight, the infusion of the tracer was disrupted and a 3-day reduction phase was made part of the experiment. The N-metabolism parameters such as protein synthesis and decomposition quotas were calculated on the basis of the temporal development of 15N-amounts excreted in urine. The labelling and reduction phases were evaluated simultaneously. By means of including the total N-amount of the animal body the balance calculation becomes more stable. The possible influence of the tracer amino acid in connection with the 3-pool-model is discussed on the basis of the ascertained parameters.
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