The objective of the present study was, to investigate the effect of varying conditions of differential centrifugation of duodenal content on the isolation of bacteria (B-fraction) and feed particles + protozoa (FP-fraction). The treatments at low-speed centrifugation were as follows: 100 x g/5 min, 400 x g/10 min, 1000 x g/10 min and 2000 x g/10 min, high speed conditions were 30,000 x g/30 min/4 degrees C. The results of three experiments are given. Analytical examination gave similar results for N-contents for all treatments, the mean values being 7.90 +/- 0.27% (n = 12) for B-fractions and 6.53 +/- 0.73% (n = 12) for FP-fractions. Increasing the low-speed from 100 x g to 2000 x g lead to increasing DAP-contents and decreasing N:DAP-ratios of the bacterial isolates, the values being 2.43, 3.02, 3.22 and 3.39 mg DAP/g DM and 32.0, 27.4, 25.0 and 23.0 N:DAP-ratio. Decreased isolation of bacterial material in the B-fraction in conjunction with increased incorporation in the FP-fraction resulted in rising the speed of the low speed centrifugation. The rates of loss of DAP, measured by comparison with the total amount were 10, 32, 48 and 70% respectively. It was concluded to prefer the isolation of bacteria from rumen fluid.

The interrelation between the time of the withdrawal of rumen fluid and the DPA: N-relation in the fraction of mixed rumen bacteria was ascertained in a test with two cows with rumen fistulae. At the moment of feeding the DPA-content of 63 mg per g bacteria was very high. In the period of 3 to 9 hours after feeding it remained relatively constant (50 mg). A mean conversion relation of 20.4 of DPA to bacteria N was ascertained. The amino acid model proved to be the same in all fractions, the content values were comparable to those found in technical literature.

Two lactating cows with duodenal re-entrance cannulae were fed with rations containing 180g urea (A) resp. urea modified with the urase inhibitor phosphoric phenyl ester diamide (PPD, 1 g PPD/100 g N). The cows, accustomed to urea, were not adapted to PPD (B) resp. had been adapted to PPD for 30 days (C). From A to C the digestibility of the organic matter in the stomachs amounted to 52, 47 and 52% and in the total tract to 78, 77 and 79%; the digestibility of crude cellulose amounted to 64, 50 and 62% resp. 69, 64 and 68% and that of starch to 91, 87 and 95% resp. 100% in each case. 71, 66 and 70% of the N were digested. The passage rate of N in the duodenum was 90, 102 and 106% of the N-intake. In the intestines 61, 68 and 76% of the N-intake were absorbed. The passage of bacteria-N in the duodenum was 67, 92 and 71% of the N-intake, that of the sum of NH3- and urea-N 6,8 and 8%. The conclusion is that, with unchanged digestibility of darbydrates, the N-supply of cows should be improved by the modification of urea with PPD after the adaptation of the cows. Cellulose fermentation is inhibited at the beginning of the adaptation to PPD.

In experiments with dairy cows provided with rumen fistulas and duodenal re-entrant canulas, the order of feeding roughage (chopped forage and pellets) and concentrates (maize and barley) was found to influence carbohydrate digestion and bacterial protein synthesis in the rumen. Feeding the roughage 90 min. before the concentrate caused a wider C2:C3 ratio and a sustained course of the pH-value (greater than 6.5) than the ratio other way round. This favoured cellulose digestion in the rumen when feeding chopped roughage. When feeding roughage before concentrates, the nonbacterial N-proportion in the duodenal contents proved greater than in the reverse order and revealed the highest values for roughage fed in pelleted form. Feeding structured roughage as first component of the diet with concentrates given afterwards is, therefore, recommended for dairy cows.

The utilisation of feed urea in the rumen was tested in 2 experiments with a total of 4 newly lactating dairy cows (13 . . . 15 and 17 . . . 19 kg resp. milk/animal and day) with rumen and duodenal re-entrant cannulae. With the energy supply remaining constant in each case, the rations in experiment A contained 8.7, 12.4 and 14.6 and those of experiment B 10.7, 13,7 and 17.1% crude plant protein in the dry matter. After the supplementation with 120 and 150 g resp. urea/animal and day there were 11.9, 15.7 and 17.8 (A) and 13.8, 16.7 and 20.2 (B) % resp. crude protein in the dry matter. The rations consisted of maize silage and a pelleted mixture of straw and concentrated feed (A) resp. maize silage, alfalfa hay and concentrated feed (B). They contained 10.3 . . . 10.6 (A) and 13.6 (B) kg dry matter with 5.6 . . . 6.0 (A) and 8.2 (B). With the increase of the crude protein level of the ration to 16.7 . . . 17.8, the absolute amount of non-NH3-N (NAN) in the duodenum increased as well. Between N-intake (g/d, x) and NAN-passage corrected by the amount of the endogenous quota (g/d, y) the relation y = 87.3 + 0.55 x (r = 0.80) could be established. NAN-passage (y) as related to N-intake decreased with the N-concentration in the dry matter of the ration (x) according to the equation y = 0.35 + 1.22x-1 (r = 0.57). 70, 62 and 61% (experiment A) and 55, 61 and 51% (experiment B) of the consumed amount of N were apparently absorbed in the intestines as NAN (without endogenous quota). The bacterial N-yield of the rumen (g, y), determined with diamino pimelic acid as microbe marker, was dependent on the consumed digestible organic matter (g, x) as follows: y = 67.3 + 0.021x (r = 0.69). There was no connection with the level of N-supply. The measuring results of the bacterial N-yield show that the utilisation rate of the urea-N decreased rapidly when there was more than 11 . . . 12% crude plant protein in the dry matter of the ration. For the tested ration type (570 . . . 600 EFUcattle/kg dry matter) the urea utilisation potential in the rumen for crude plant protein concentrations of 8.7, 10.7, 12.4, 13.4, 13.7, 14.6 and 17.1% in the dry matter was 13.0, 6.9, 4.1, 2.6, 1.3 and -9.6 g urea/kg dry matter.