Forage brassicas, such as summer turnip (ST; Brassica rapa) and forage rape (FR; Brassica napus), are used as supplementary crops during summer. However, studies with lactating dairy cows fed these forages are limited and report inconsistent productive responses. The aim of this study was to determine dry matter intake, rumen fermentation and milk production responses of dairy cows in mid-lactation supplemented with and without summer (‘ST’ or ‘FR’) brassicas. Twelve multiparous lactating dairy cows were randomly allocated to three dietary treatments in a replicated 3 × 3 Latin square design balanced for residual effects over three 21-day periods. The control diet consisted of 16.2 kg DM of grass silage, 2.25 kg DM of commercial concentrate and 2.25 kg DM solvent-extracted soybean meal. For the other two dietary treatments, 25% of the amounts of silage and concentrates were replaced with FR or ST. The inclusion of forage brassicas had no effects on milk production (24.2 kg cow/day average) and composition (average milk fat and protein 43.2 and 33.6 g/l, respectively). Dry matter intake was 0.98 kg and 1.12 kg lower for cows supplemented with FR and ST, respectively, resulting in a greater feed conversion efficiency (1.35 kg milk/kg DM for ST and FR v. 1.27 kg milk/kg DM for the control diet). Intraruminal pH was lower for cows supplemented with ST compared to the control diet; however, it did not decrease below pH 5.8 at any time of the day. After feeding, the concentrations of total short-chain fatty acids (SCFAs) in rumen contents increased with ST supplementation compared to the control diet. Inclusion of FR in the diet increased the molar proportion of acetate (68.5 mmol/100 mmol) in total SCFA at the expense of propionate, measured 6 h after feeding of the forage. The molar proportion of butyric acid was greater with ST and FR supplementation (13.1 and 12 mmol/100 mmol, respectively) than in control cows. The estimated microbial nitrogen (N) flow was 89.1 g/day greater when supplementing FR compared to the control diet. Based on the haematological measures, the inclusion of summer brassica forages did not affect the health status of the animals. These results indicate that mid-lactation dairy cows fed brassicas are able to maintain production despite the reduced intake, probably due to improved rumen fermentation and therefore nutrient utilization.

The combined addition of branched-chain volatile fatty acids (BCVFAs) and folic acid (FA) could improve growth performance and nutrient utilization by stimulating ruminal microbial growth and enzyme activity. This study was conducted to evaluate the effects of BCVFA and FA addition on growth performance, ruminal fermentation, nutrient digestibility, microbial enzyme activity, microflora and excretion of urinary purine derivatives (PDs) in calves. Thirty-six Chinese Holstein weaned calves (60 ± 5.4 days of age and 107 ± 4.7 kg of BW) were assigned to one of four groups in a randomized block design. Treatments were control (without additives), FA (with 10 mg FA/kg dietary DM), BCVFA (with 5 g BCVFA/kg dietary DM) and the combined addition of FA and BCVFA (10 mg/kg DM of FA and 5 g/kg DM of BCVFA). Supplements were hand-mixed into the top one-third of total mixed ration. Dietary concentrate to maize silage ratio was 50 : 50 on a DM basis. Dietary BCVFA or FA addition did not affect dry matter intake but increased average daily gain (ADG) and feed conversion efficiency. Ruminal pH and ammonia N were lower, and total volatile fatty acids (VFAs) concentration was higher for BCVFA or FA addition than for control. Dietary BCVFA or FA addition did not affect acetate proportion but decreased propionate proportion and increased acetate to propionate ratio. Total tract digestibility of DM, organic matter, CP and NDF was higher for BCVFA or FA addition than for control. Dietary BCVFA or FA addition increased activity of carboxymethyl cellulase and cellobiase, population of total bacteria, fungi, Ruminococcus albus, R. flavefaciens, Fibrobacter succinogenes and Prevotella ruminicola as well as total PD excretion. Ruminal xylanase, pectinase and protease activity and Butyrivibrio fibrisolvens population were increased by BCVFA addition, whereas population of protozoa and methanogens was increased by FA addition. The BCVFA × FA interaction was significant for acetate to propionate ratio, cellobiase activity and total PD excretion, and these variables increased more with FA addition in diet without BCVFA than in diet with BCVFA. The data indicated that supplementation with BCVFA or FA increased ADG, nutrient digestibility, ruminal total VFA concentration and microbial protein synthesis by stimulating ruminal microbial growth and enzyme activity in calves.

The objective was to investigate the effect of variation in forage source and feed particle size of a diet, including interactions, on the amount and the composition of microbial crude protein (CP) in a semi-continuous culture system (Rusitec). Different microbial CP fractions were compared. Five diets with mean forage proportion of 0.88 and different maize silage to grass silage ratios (100 : 0, 79 : 21, 52 : 48, 24 : 76 and 0 : 100) were used. Diets were ground through sieves with a pore size of either 1 or 4 mm, matching the particle size of fine (F) and coarse (C), respectively. Diets were characterised by increasing concentrations of CP and fibre fractions, and decreasing concentrations of starch with ascending inclusion rates of grass silage. Microbial mass was isolated from feed residues after incubation from the liquid phase of the fermenter and from the liquid effluent. The amount of synthesised microbial CP was determined on the basis of 15N balance. It increased quite linearly by the stepwise replacement of maize silage by grass silage, and was higher in C treatments compared to F treatments. Efficiency of microbial CP synthesis (EMPS) was improved from 29 to 43 mg microbial N/g degraded organic matter (OM) by increasing the proportion of grass silage in the diet, but was unaffected by particle size. The N content as well as the profiles of amino acids of the three microbial fractions was affected by diet composition and particle size. The ratio of solid- to liquid-associated microbes was affected by diet composition and feed particle size. The amount and EMPS seemed to be improved by degradation of OM from grass silage and an increasing availability of N. Moreover, the results of this study indicated a shift in the composition of the microbial community caused by variation in forage composition and feed particle size.

Three groups of five wethers with ruminal and duodenal cannulas and maintained as either fauna-free (FF) or inoculated with total mixed fauna (TF) or Entodinium caudatum as a single-species monofauna (EN) were used in an experiment with two 28 d periods. In the first period, the sheep were fed a control barley-based diet (40:60 concentrate to silage DM) and in the second period the diet was supplemented with 187 g DM of Enterolobium cyclocarpum for the last 12 d of the period. The diets of period 1 and 2 were isonitrogenous. There was no effect of fauna on apparent ruminal and total tract organic matter and fibre digestion, but bacterial and microbial N flow and efficiency were improved in FF sheep compared to TF sheep. In period 2, protozoal numbers were reduced between 31 and 88 % 2 h after feeding E. cyclocarpum for the third to twelfth day of supplementation and by an average of 25 % in samples collected over the 24 h feeding cycle. Supplementation of the diet with E. cyclocarpum and the consequent protozoal reduction in TF and EN sheep improved the flow of non-ammonia N and bacterial N to the small intestine and the efficiency of microbial synthesis. However, E. cyclocarpum reduced ruminal organic matter digestion, especially in faunated sheep, and total tract organic matter, N and fibre digestion. Thus, a reduction in the protozoal cell numbers of 25 % was sufficient to achieve the beneficial effects of reduced fauna on the bacterial protein supply, but diet digestibility was reduced.

Five experiments were carried out to extend knowledge of purine metabolism in the camel (Camelus dromedarius) and to establish a model to enable microbial protein outflow from the forestomachs to be estimated from the urinary excretion of purine derivatives (PD; i.e. xanthine, hypoxanthine, uric acid, allantoin). In experiment 1, four camels were fasted for five consecutive days to enable endogenous PD excretion in urine to be determined. Total PD excretion decreased during the fasting period to 267 (se 41·5)?μmol/kg body weight (W)0·75 per d. Allantoin and xanthine+hypoxanthine were consistently 86 and 6·1?% of total urinary PD during this period but uric acid increased from 3·6?% to 7·4?%. Xanthine oxidase activity in tissues (experiment 2) was (μmol/min per g fresh tissue) 0·038 in liver and 0·005 in gut mucosa but was not detected in plasma. In experiment 3, the duodenal supply of yeast containing exogenous purines produced a linear increase in urinary PD excretion rate with the slope indicating that 0·63 was excreted in urine. After taking account of endogenous PD excretion, the relationship can be used to predict purine outflow from the rumen. From the latter prediction, and also the purine:protein ratio in bacteria determined in experiment 5, we predicted the net microbial outflow from the rumen. In experiment 4, with increasing food intake, the rate of PD excretion in the urine increased linearly by about 11·1?mmol PD/kg digestible organic matter intake (DOMI), equivalent to 95?g microbial protein/kg DOMI.

The first objective of this experiment was to investigate the effect of using different bacterial isolates on the estimation of microbial protein production in semi-continuous fermenters (RUSITEC) given four diets, and to test whether a ‘mixed’ bacterial pellet was representative of the whole bacterial population in the fermenters. A second objective was to compare two different microbial markers (nitrogen-fifteen (15N) and purine bases (PB)). Diets consisted of neutral-detergent fibre from grass hay (10 g/day) and sugar-beet pulp (2 g/day) and 280 mg/day of one of four N forms (isolated soya-bean protein, soya-bean peptides, amino acids blended to profile soya-bean protein and NH4Cl). Two 14-day incubation runs were carried out and in each run each of the four different diets were given to two vessels. On days 12 and 13, total digesta (effluent plus nylon bags residues) was collected for analyses of non-ammonia N, 15N enrichment and PB concentration, and for isolation of total mixed bacterial pellets (TB). On the last day of each incubation run, the system was stopped for isolation of liquid- (LAB) and solid-associated (SAB) bacteria. Microbial N flow was estimated from the 15N enrichment and PB concentration in both total digesta and in the three different bacterial pellets (TB, LAB, and SAB). For all diets, LAB presented a greater (P < 005) 15N enrichment and PB: N ratio than SAB, with TB having an intermediate value. For both markers, the use of LAB produced the lowest (P < 005) estimates of microbial N flow and the use of SAB produced the greatest (P < 005) estimates. The use of TB produced intermediate values with all diets, suggesting that TB consisted of SAB and LAB. For all bacterial pellets, PB produced greater (P 005) values of microbial N flow than 15N. However, there was a positive relationship (r = 0·883; P 0001; no. = 15) between the values of microbial N flow determined with the two markers when TB were used as reference.

The kinetics of allantoin metabolism were studied in rumen-cannulated sheep by means of a single intravenous injection of [4,5-14C]allantoin. The decline in the specific radioactivity of allantoin in plasma following the injection of tracer was best described by a double exponential function, indicating that allantoin moves in and out of two or more kinetically distinct compartments. Sequestering of tracer in secondary or tertiary compartments in the body water appears likely to have resulted in overestimation of net flux of allantoin through the blood in the present study. In future studies, sampling of blood for several days after administration of tracer should alleviate this problem. About 80 % of the [14C]allantoin injected was recovered as [14C]allantoin in urine during the 12 h after tracer injection, increasing to 94 % after 4 d. Allantoin-C also passed through the blood bicarbonate pool, suggesting that allantoin is degraded in the gastrointestinal tract. A small amount of allantoin-C (4 % of the net flux of allantoin through the blood pool) was apparently degraded to form bicarbonate-C in the rumen and postruminally, and subsequently appeared in blood bicarbonate-C. Transfer of allantoin-C into the rumen via saliva was insignificant. In view of these findings, the net flux of allantoin through the blood should be a better predictor of rumen microbial outflow than urinary allantoin excretion, because urinary excretion of purine derivatives must be adjusted for conversion of allantoin-C to blood bicarbonate when used to predict the flow of microbial protein from the rumen.

The potential of milk allantoin as an index ofmicrobial protein supply was evaluated in two experiments conducted with 12 multiparous Holstein-Friesian dairy cows that examined the effects of altering the supply of metabolizable energy (ME) and fermentable metabolizable energy (FME). In the first experiment, late lactation cows received a 14·3 kg dry matter (DM) per day basal diet consisting of (g/kg DM) chopped barley straw (415), soya-bean meal (322), molassed sugar-beet pulp (197), molasses (33), urea (17) and a vitamin and mineral supplement (17) for a 21-day co-variance period. During four 16-day periods, six treatments were allocated randomly to cows, consisting of potato starch (1, 2 or 3 kg DM per day) or fat supplements (0·64, 1·27 or 1·91 kg DM per day). In the second experiment, early lactation cows received 40 kg/day (fresh weight, (FW)) of silage (307 g/kg toluene-corrected dry matter, 149 g/kg DM crude protein and 11·6 MJ/kg ME). During three 21-day periods, four treatments were evaluated consisting of supplements of either 4·1 (LI) or 8·1 (12) kg fresh weight per day of a low fat concentrate (acid hydrolysis ether extract (AHEE) 45 g/kg DM) or 3·8 (HI) or 7·5 (H2) kg fresh weight per day of a high fat concentrate (AHEE 110 g/kg DM). Both experiments showed individual cow milk allantoin concentration or excretion to be poorly correlated with urinary purine derivative excretion or calculated microbial protein supply. Use of treatment mean (TM) values dramatically improved these relationships. For pooled TM (no. = 10) values from both experiments, close relationships existed between milk allantoin excretion and concentration with milk yield (r values 0·991 and 0·883, respectively). Auto-correlation with milk yield appeared to account for milk allantoin excretion and concentration being highly correlated with urinary purine derivative excretion (r values 0·908 and 0·934, respectively) and calculated microbial protein supply (r values 0·938 and 0·945, respectively). Current experimental data indicates that measurement of milk allantoin is not a reliable indicator of microbial protein supply for individual cows.

Four sheep (live weight 39 to 42 kg) fitted with rumen cannulas were offered ammonia-treated barley straw alone (control) or supplemented with unmolassed sugar-beet pulp or rolled barley at 0·20 or 0·40 of the total diet on a fresh weight basis (SBP20, SBP40, B20 and B40, respectively) using a 4 × 5 randomized block design. Total diet intakes were 710, 873, 1054, 843 and 1021 g dry matter per day for control, SBP20, SBP40, B20 and B40 respectively. The intake of straw was depressed significantly with SBP40 and B40 (P < 0·05). Rumen pH and ammonia concentration decreased, and total volatile fatty acid concentrations increased (P < 0·05) with increasing level of supplementation for both supplements. Rumen liquid outflow rate was affected little by the supplementation although this measurement for B40 was significantly higher than that for the other treatments. Microbial protein supply, estimated from urinary excretion of purine derivatives, increased significantly with the increasing amounts of supplements (4·5, 6·7, 8·5, 7·1 and 9·0 (s.e.d. 0·73) g N per day for control, SBP20, SBP40, B20 and B40 respectively) (P < 0·05). The calculated efficiency of microbial protein production was 12·8, 14·2, 14·3, 15·7 and 14·5 (s.e.d. 1·45) g N per kg digestible organic matter intake (DOMI) for the five treatments. Although this value was lowest with the control, differences between treatments were not significant (P > 0·05). It seems that microbial protein yield per unit DOMI for a diet based on ammonia-treated straw could not be improved substantially by supplementation with sugar-beet pulp or barley.

The diurnal changes in plasma concentration and urinary excretion of purine derivatives (PD) were examined in four steers given their food in either one or two meals daily in two 18-day periods. In period 1, two steers were fed once daily (at 08.00 h) and the other two twice daily (at 08.00 and 16.00 h). In period 2, the treatments were switched. In the last 4 days of each period, plasma samples were taken from 08.00 to 24.00 h at 2 to 3 h intervals, and daily urine was collected in several 4 to 8h periods. With once daily feeding, allantoin concentration in plasma varied from 090 to 1·07 of the daily mean and with twice daily feeding, from 0·97 to 1·05 of mean. The coefficient of variation (CV) for total variation was 0·14. In urine, the concentration and hourly excretion of PD varied with sampling times (P < 0·05) but the ratio of PD to creatinine was relatively constant even for once-daily feeding (CV, 0·02 for measurements at different times). The CV for total variation in the ratio was 0·19. The results suggest that diurnal variation in plasma allantoin concentration and in the ratio of allantoin (or total PD) to creatinine are relatively small even for once daily feeding. It thus seems that these measurements in spot samples may be suitable for use as a simple index of microbial protein supply if they are correlated with the daily PD excretion. The results also indicate that measurements in samples of plasma and urine taken between 15.00 and 20.00 h tended to give a better representation of the daily mean.