Comparative metabolome analysis of ruminal changes in Holstein dairy cows fed low- or high-concentrate diets

Introduction

Currently, information on the comprehensive changes in the ruminal metabolites of dairy cows fed high-concentrate diet is limited.

Objectives

This study aimed to compare the composition of whole-ruminal metabolites in dairy cows that were fed a low concentrate diet or a high concentrate diet using modern metabolome analysis.

Methods

Cows were fed a low-concentrate diet (LC; 40% concentrate feeds, dry matter (DM) basis) or a high-concentrate diet (HC; 70% concentrate feeds, DM basis). GC/MS was used to analyze rumen fluid samples.

Results

As compared with the LC group, HC diet significantly increased the concentration of bacterial degradation products (included xanthine, hypoxanthine, uracil, etc.), some toxic compounds (included lipopolysaccharide, biogenic amines, ethanolamine, etc.) and 15 amino acids (included alanine, leucine, glycine, etc.). The enrichment analysis of differentially expressed metabolites indicated that three pathways, including aminoacyl-tRNA biosynthesis; phenylalanine, tyrosine, and tryptophan biosynthesis; and valine, leucine and isoleucine biosynthesis, were significantly enriched after the diet treatments. Correlation network analysis revealed that HC diets altered the ruminal metabolic pattern, and the metabolites in the HC group were more complicated than those in the LC group. The correlations between ruminal metabolites and blood parameters were mainly centralized in the ruminal metabolites and albumin (40 metabolites), followed by globulin (18 metabolites) and total protein (6 metabolites).

Conclusions

These findings revealed that HC feeding altered the concentrations of ruminal metabolites as well as the metabolic pattern, and the rumen metabolism could be reflected by blood metabolism to a certain degree.

     

Contribution of dietary arginine to nitrogen utilisation and excretion in juvenile sea bass (Dicentrarchus labrax) fed diets differing in protein source

The role of dietary arginine in affecting nitrogen utilisation and excretion was studied in juvenile European sea bass (Dicentrarchus labrax) fed for 72 days with diets differing in protein sources (plant protein-based (PM) and fish-meal-based (FM)). Fish growth performance and nitrogen utilisation revealed that dietary Arg surplus was beneficial only in PM diets. Dietary Arg level significantly affected postprandial plasma urea concentrations. Hepatic arginase activity increased (P<0.05) in response to dietary Arg surplus in fish fed plant protein diets; conversely ornithine transcarbamylase activity was very low and inversely related to arginine intake. No hepatic carbamoyl phosphate synthetase III activity was detected. Dietary arginine levels did not affect glutamate dehydrogenase activity. A strong linear relationship was found between liver arginase activity and daily urea-N excretion. Dietary Arg excess reduced the proportion of total ammonia nitrogen excreted and increased the contribution of urea-N over the total N excretion irrespective of dietary protein source. Plasma and excretion data combined with enzyme activities suggest that dietary Arg degradation via hepatic arginase is a major pathway for ureagenesis and that ornithine-urea cycle is not completely functional in juvenile sea bass liver.     

Effects of dietary crude protein concentration on animal performance and nitrogen utilisation efficiency at different stages of lactation in Holstein-Friesian dairy cows

Nitrogen (N) excretion from livestock production systems is of significant environmental concern; however, few studies have investigated the effect of dietary CP concentration on N utilisation efficiency at different stages of lactation, and the interaction between dietary CP levels and stages of lactation on N utilisation. Holstein-Friesian dairy cows (12 primiparous and 12 multiparous) used in the present study were selected from a larger group of cows involved in a whole-lactation study designed to examine the effect of dietary CP concentration on milk production and N excretion rates at different stages of lactation. The total diet CP concentrations evaluated were 114 (low CP), 144 (medium CP) and 173 (high CP) g/kg DM, with diets containing (g/kg DM) 550 concentrates, 270 grass silage and 180 maize silage. During early (70–80 days), mid- (150–160 days) and late (230–240 days) lactation, the same 24 animals were transferred from the main cow house to metabolism units for measurements of feed intake, milk production and faeces and urine outputs. Diet had no effect on BW, body condition score, or milk fat, protein or lactose concentration, but DM intake, milk yield and digestibilities of DM, energy and N increased with increasing diet CP concentration. The effect of diet on milk yield was largely due to differences between the low and medium CP diets. Increasing dietary CP concentration significantly increased urine N/N intake and urine N/manure N, and decreased faecal N/N intake, milk N/N intake and manure N/N intake. Although increasing dietary CP level significantly increased urine N/milk yield and manure N/milk yield, differences in these two variables between low and medium CP diets were not significant. There was no significant interaction between CP level and stage of lactation on any N utilisation variable, indicating that the effects of CP concentration on these variables were similar between stages of lactation. These results demonstrated that a decrease in dietary CP concentration from high (173 g/kg DM) to medium level (144 g/kg DM) may be appropriate for Holstein-Friesian dairy cow to maintain milk production efficiency, whilst reducing both urine N and manure N as a proportion of N intake or milk production.     

Comparative analysis of gene expression profiles in ruminal tissue from Holstein dairy cows fed high or low concentrate diets

Cattle are often fed high concentrate (HC) diets to increase productivity, although HC diets cause changes in ruminal environment such as pH reduction. Despite those well-documented changes in cattle fed HC diets, there is currently a paucity of data describing the molecular events regulating the ruminal environment. Our objective was to gain an understanding of which genes are differentially expressed in ruminal tissue from Holstein cows fed a HC comparing to low concentrate (LC) diet using microarray analysis using a bovine 24 k microarray. A total of 5,200 differentially expressed genes (DEG) were detected for cows fed HC relative to LC. The DEG were firstly annotated with gene ontology (GO) and Kyoto encyclopedia of Genes and Genomes (KEGG), indicating that the DEG were associated with catalytic activity and MAPK pathway, respectively. Further characterization using GeneCodis identified patterns of interrelated annotations for the DEG to elucidate the relationships among annotation groups revealed that a cAMP-dependent protein kinase A catalytic subunit beta (PRKACB), may be associated with ruminal tissue maintenance. The results contributed to understanding of the regulatory mechanisms at the mRNA level for Holstein cows fed at different concentrate ratio diets.     

Effect of niacin supplementation on digestibility,nitrogen utilisation and milk and blood variables in lactating dairy cows fed a diet with a negative rumen nitrogen balance

The aim of the present experiment was to determine if a niacin supplementation of 6 g/d to lactating dairy cow diets can compensate negative effects of a rumen nitrogen balance (RNB) deficit. A total of nine ruminally and duodenally fistulated lactating multiparous German Holstein cows were successively assigned to one of three diets consisting of 10 kg maize silage (dry matter [DM] basis) and7 kg DM concentrate: Diet RNB– (n = 6) with energy and utilisable crude protein at the duodenum (uCP) according to the average requirement of the animals but with a negative RNB (–0.41 g N/MJ metabolisable energy [ME]); Diet RNB0 (n = 7) with energy, uCP and a RNB (0.08 g N/MJ ME) according to the average requirement of the animals and, finally, Diet NA (n = 5), which was the same diet as RNB–, but supplemented with 6 g niacin/d. Samples of milk were taken on two consecutive days, blood samples were taken on one day pre- and post-feeding and faeces and urine were collected completely over five consecutive days. The negative RNB reduced milk and blood urea content and apparent total tract digestibility of DM, organic matter (OM) and neutral detergent fibre (NDF). Also N excretion with urine, the total N excreted with urine and faeces and the N balance were reduced when the RNB was negative. Supplementation of niacin elevated plasma glucose concentration after feeding and the N balance increased. Supplementing the diet with a negative RNB with niacin led to a more efficient use of dietary N thereby avoiding the negative effects of the negative RNB on the digestibility of DM, OM and NDF.     

Effect of niacin supplementation on digestibility, nitrogen utilisation and milk and blood variables in lactating dairy cows fed a diet with a negative rumen nitrogen balance

The aim of the present experiment was to determine if a niacin supplementation of 6 g/d to lactating dairy cow diets can compensate negative effects of a rumen nitrogen balance (RNB) deficit. A total of nine ruminally and duodenally fistulated lactating multiparous German Holstein cows were successively assigned to one of three diets consisting of 10 kg maize silage (dry matter [DM] basis) and 7 kg DM concentrate: Diet RNB- (n = 6) with energy and utilisable crude protein at the duodenum (uCP) according to the average requirement of the animals but with a negative RNB (-0.41 g N/MJ metabolisable energy [ME]); Diet RNB0 (n = 7) with energy, uCP and a RNB (0.08 g N/MJ ME) according to the average requirement of the animals and, finally, Diet NA (n = 5), which was the same diet as RNB-, but supplemented with 6 g niacin/d. Samples of milk were taken on two consecutive days, blood samples were taken on one day pre- and post-feeding and faeces and urine were collected completely over five consecutive days. The negative RNB reduced milk and blood urea content and apparent total tract digestibility of DM, organic matter (OM) and neutral detergent fibre (NDF). Also N excretion with urine, the total N excreted with urine and faeces and the N balance were reduced when the RNB was negative. Supplementation of niacin elevated plasma glucose concentration after feeding and the N balance increased. Supplementing the diet with a negative RNB with niacin led to a more efficient use of dietary N thereby avoiding the negative effects of the negative RNB on the digestibility of DM, OM and NDF.     

Transient reductions in milk fat synthesis and their association with the ruminal and metabolic profile in dairy cows fed high-starch,low-fat diets

Sub-acute ruminal acidosis (SARA) is sometimes observed along with reduced milk fat synthesis. Inconsistent responses may be explained by dietary fat levels. Twelve ruminally cannulated cows were used in a Latin square design investigating the timing of metabolic and milk fat changes during Induction and Recovery from SARA by altering starch levels in low-fat diets. Treatments were (1) SARA Induction, (2) Recovery and (3) Control. Sub-acute ruminal acidosis was induced by feeding a diet containing 29.4% starch, 24.0% NDF and 2.8% fatty acids (FAs), whereas the Recovery and Control diets contained 19.9% starch, 31.0% NDF and 2.6% FA. Relative to Control, DM intake (DMI) and milk yield were higher in SARA from days 14 to 21 and from days 10 to 21, respectively (P < 0.05). Milk fat content was reduced from days 3 to 14 in SARA (P < 0.05) compared with Control, while greater protein and lactose contents were observed from days 14 to 21 and 3 to 21, respectively (P < 0.05). Milk fat yield was reduced by SARA on day 3 (P < 0.05), whereas both protein and lactose yields were higher on days 14 and 21 (P < 0.05). The ruminal acetate-to-propionate ratio was lower, and the concentrations of propionate and lactate were higher in the SARA treatment compared with Control on day 21 (P < 0.05). Plasma insulin increased during SARA, whereas plasma non-esterified fatty acids and milk β-hydroxybutyrate decreased (P < 0.05). Similarly to fat yield, the yield of milk preformed FA (>16C) was lower on day 3 (P < 0.05) and tended to be lower on day 7 in SARA cows (P < 0.10), whereas yield of de novo FA (<16C) was higher on day 21 (P < 0.01) in the SARA group relative to Control. The t10- to t11-18:1 ratio increased during the SARA Induction period (P < 0.05), but the concentration of t10-18:1 remained below 0.5% of milk fat, and t10,c12 conjugated linoleic acid remained below detection levels. Odd-chain FA increased, whereas branched-chain FA was reduced during SARA Induction from days 3 to 21 (P < 0.05). Sub-acute ruminal acidosis reduced milk fat synthesis transiently. Such reduction was not associated with ruminal biohydrogenation intermediates but rather with a transient reduction in supply of preformed FA. Subsequent rescue of milk fat synthesis may be associated with higher availability of substrates due to increased DMI during SARA.     

Dietary carbohydrate composition modifies the milk N efficiency in late lactation cows fed low crude protein diets

Nitrogen emissions from dairy cows can be readily decreased by lowering the dietary CP concentration. The main objective of this work was to test whether the milk protein yield reduction associated with low N intakes could be partially compensated for by modifying the dietary carbohydrate composition (CHO). The effects of CHO on digestion, milk N efficiency (milk N/N intake; MNE) and animal performance were studied in four Jersey cows fed 100% or 80% of the recommended protein requirements using a 4×4 Latin square design. Four iso-energetic diets were formulated to two different CHO sources (starch diets with starch content of 34.3% and NDF at 32.5%, and fiber diets with starch content of 5.5% and NDF at 49.1%) and two CP levels (Low=12.0% and Normal=16.5%). The apparent digestible organic matter intake (DOMI) and the protein supply (protein digestible in the small intestine; PDIE) were similar between starch and fiber diets. As planned, microbial N flow (MNF) to the duodenum, estimated from the urinary purine derivatives (PD) excretion, was similar between Low and Normal CP diets. However, the MNF and the efficiency of microbial synthesis (g of microbial N/kg apparently DOMI) were higher for starch v. fiber diets. Milk and milk N fractions (CP, true protein, non-protein N (NPN)) yield were higher for starch compared with fiber diets and for Normal v. Low CP diets. Fecal N excretion was similar across dietary treatments. Despite a higher milk N ouput with starch v. fiber diets, the CHO modified neither the urinary N excretion nor the milk urea-N (MUN) concentration. The milk protein yield relative to both N and PDIE intakes was improved with starch compared with fiber diets. Concentrations of β-hydroxybutyrate, urea and Glu increased and those of glucose and Ala decreased in plasma of cows fed starch v. fiber diets. On the other hand, plasma concentration of albumin, urea, insulin and His increased in cows fed Normal compared with Low CP diets. This study showed that decreasing the dietary CP proportion from 16.5% to 12.0% increases and decreases considerably the MNE and the urinary N excretion, respectively. Moreover, present results show that at similar digestible OM and PDIE intakes, diets rich in starch improves the MNE and could partially compensate for the negative effects of Low CP diets on milk protein yield.     

Milk yield response of dairy cows fed fat along with protein

The influence of a fat-coated protein on milk production of Holstein dairy cows was determined using a 4×4 Latin square experiment. Twelve cows were fed a control diet or test diets supplemented with fat, fat plus ruminally undegraded protein (RUP), or a fat-coated protein (Duets™). Cows fed test diets received 0.55 kg of more fat per day than cows in the control treatment. Daily intakes of feed, energy, and protein were the same in all treatments. Cows produced 36.5, 37.3, 37.9, and 39.3 kg of energy-corrected milk per day in control, fat, fat plus RUP, and fat-coated protein treatments, respectively. Cows fed fat-supplemented diets produced an average 1.7 kg more milk daily compared with cows in the control treatment. Feeding RUP along with fat or fat-coated protein provided no further improvement in milk yield compared with fat alone, but partially alleviated the depression in protein content caused by supplemental fat and increased the daily yield of milk protein. In the present experiment, cows fed fat-coated protein produced daily an average 60 g of milk protein more than cows fed fat alone. Since, there was no advantage in milk yield, the decision to include fat-coated protein in dairy rations should be based on its price compared to fat alone and the return in terms of milk protein yield.     

Relationship between rumen methanogens and methane production in dairy cows fed diets supplemented with a feed enzyme additive

Aims: To investigate the relationship between ruminal methanogen community and host enteric methane (CH₄) production in lactating dairy cows fed diets supplemented with an exogenous fibrolytic enzyme additive. Methods and Results: Ecology of ruminal methanogens from dairy cows fed with or without exogenous fibrolytic enzymes was examined using PCR–denaturing gradient gel electrophoresis (PCR–DGGE) analyses and quantitative real‐time PCR (qRT‐PCR). The density of methanogens was not affected by the enzyme additive or sampling times, and no relationship was observed between the total methanogen population and CH₄ yield (as g per head per day or g kg^?1 DMI). The PCR–DGGE profiles consisted of 26 distinctive bands, with two bands similar to Methanogenic archaeon CH1270 negatively correlated, and one band similar to Methanobrevibacter gottschalkii strain HO positively correlated, with CH₄ yield. Three bands similar to Methanogenic archaeon CH1270 or Methanobrevibacter smithii ATCC 35061 appeared after enzyme was added. Conclusions: Supplementing a dairy cow diet with an exogenous fibrolytic enzyme additive increased CH₄ yield and altered the composition of the rumen methanogen community, but not the overall density of methanogens. Significance and Impact of the Study: This is the first study to identify the correlation between methanogen ecology and host CH₄ yield from lactating dairy cows.     

Characterisation of particle dynamics and turnover in the gastrointestinal tract of Holstein cows fed forage diets differing in fibre and protein contents

An improved understanding of the role of forage quality on the processes of particle dynamics and turnover is important for the development of healthier and cost-effective feeding strategies that aim at lowering the proportions of concentrates in the diets of cattle. The aim of this study was to evaluate the effects of feeding hays of different qualities on particle dynamics, digestion kinetics and turnover in the gastrointestinal tract (GIT). Three non-lactating, rumen fistulated Holstein cows were fed diets consisting exclusively of hay with either low quality [Group LH; 605 ± 12.4 g/kg neutral detergent fibre (NDF) and 63 ± 4.7 g/kg crude protein (CP)] or good quality (Group GH; 551 ± 20.1 g/kg NDF and 116 ± 3.6 g/kg CP). Data showed that in situ dry matter (DM) disappearance of the soluble fraction was greater for Group GH (p < 0.05). Feeding good quality hay also lowered the proportion of particles >1.18 mm particularly during the eating process (p < 0.05). Changes in the particle size occurring afterwards were greater for Group GH as well (p < 0.05); approximately 30% in the comminution in the particle size occurred postruminally. Feeding hay of good quality lowered DM content of solid rumen digesta (p < 0.05), accelerated (p < 0.05) the turnover rate of DM and NDF in the GIT and increased DM intake (p < 0.05). In conclusion, feeding forages of better quality significantly promoted degradation processes and kinetics in the GIT with positive effects on turnover rate of digesta and feed intake in Holstein cows.     

Characterisation of particle dynamics and turnover in the gastrointestinal tract of Holstein cows fed forage diets differing in fibre and protein contents

An improved understanding of the role of forage quality on the processes of particle dynamics and turnover is important for the development of healthier and cost-effective feeding strategies that aim at lowering the proportions of concentrates in the diets of cattle. The aim of this study was to evaluate the effects of feeding hays of different qualities on particle dynamics, digestion kinetics and turnover in the gastrointestinal tract (GIT). Three non-lactating, rumen fistulated Holstein cows were fed diets consisting exclusively of hay with either low quality [Group LH; 605 ± 12.4 g/kg neutral detergent fibre (NDF) and 63 ± 4.7 g/kg crude protein (CP)] or good quality (Group GH; 551 ± 20.1 g/kg NDF and 116 ± 3.6 g/kg CP). Data showed that in situ dry matter (DM) disappearance of the soluble fraction was greater for Group GH (p < 0.05). Feeding good quality hay also lowered the proportion of particles >1.18 mm particularly during the eating process (p < 0.05). Changes in the particle size occurring afterwards were greater for Group GH as well (p < 0.05); approximately 30% in the comminution in the particle size occurred postruminally. Feeding hay of good quality lowered DM content of solid rumen digesta (p < 0.05), accelerated (p < 0.05) the turnover rate of DM and NDF in the GIT and increased DM intake (p < 0.05). In conclusion, feeding forages of better quality significantly promoted degradation processes and kinetics in the GIT with positive effects on turnover rate of digesta and feed intake in Holstein cows.     

Quantitative analysis of ruminal bacterial populations involved in lipid metabolism in dairy cows fed different vegetable oils

Vegetable oils are used to increase energy density of dairy cow diets, although they can provoke changes in rumen bacteria populations and have repercussions on the biohydrogenation process. The aim of this study was to evaluate the effect of two sources of dietary lipids: soybean oil (SO, an unsaturated source) and hydrogenated palm oil (HPO, a saturated source) on bacterial populations and the fatty acid profile of ruminal digesta. Three non-lactating Holstein cows fitted with ruminal cannulae were used in a 3×3 Latin square design with three periods consisting of 21 days. Dietary treatments consisted of a basal diet (Control, no fat supplement) and the basal diet supplemented with SO (2.7% of dry matter (DM)) or HPO (2.7% of DM). Ruminal digesta pH, NH₃–N and volatile fatty acids were not affected by dietary treatments. Compared with control and HPO, total bacteria measured as copies of 16S ribosomal DNA/ml by quantitative PCR was decreased (P<0.05) by SO. Fibrobacter succinogenes, Butyrivibrio proteoclasticus and Anaerovibrio lipolytica loads were not affected by dietary treatments. In contrast, compared with control, load of Prevotella bryantii was increased (P<0.05) with HPO diet. Compared with control and SO, HPO decreased (P<0.05) C18:2 cis n-6 in ruminal digesta. Contents of C15:0 iso, C18:11 trans-11 and C18:2 cis-9, trans-11 were increased (P<0.05) in ruminal digesta by SO compared with control and HPO. In conclusion, supplementation of SO or HPO do not affect ruminal fermentation parameters, whereas HPO can increase load of ruminal P. bryantii. Also, results observed in our targeted bacteria may have depended on the saturation degree of dietary oils.     

N-alkanes v. ytterbium/faecal index as two methods for estimating herbage intake of dairy cows fed on diets differing in the herbage: maize silage ratio and feeding level

The aim of this study was to compare the n-alkanes and the ytterbium (Yb)/faecal index techniques as two methods for estimating the herbage intake of dairy cows fed indoors on different herbage : supplement ratios and feeding levels. The supplement was a mixture of maize silage and soyabean meal (ratio of 87 : 13 on a dry matter (DM) basis). In all, four treatments were studied. The herbage : supplement ratio in the diet was 25 : 75, 50 : 50, 75 : 25 and 50 : 50 for treatments 1, 2, 3 and 4, respectively. Animals were offered for treatments 1, 2 and 3, 100% of ad libitum intake measured before the experiment and 70% of ad libitum intake for treatment 4. Cows were fed herbage in the morning and supplement in the evening. A total of six lactating Holstein dairy cows were used in a 4 × 4 Latin square with four 14-day periods. Herbage and supplement intakes, faecal output (FO), in vivo organic matter (OM) digestibility and faecal recovery of markers were measured on the last 5 days of each period. Intake was estimated with the two methods and from two faecal sampling techniques, that is, total faecal collection v. grab sampling during milking. Mean herbage intake as fed, or estimated from n-alkanes or from the Yb/faecal index was 7.7, 8.1 and 10.2 kg DM, respectively. The mean prediction error, expressed as a fraction of actual herbage intake, was 0.10 and 0.50 for the n-alkanes and Yb/faecal index methods, respectively. The n-alkanes method clearly showed much better accuracy than the Yb/faecal index method for estimating intake, irrespective of the faecal sampling method, herbage : silage proportion or feeding level. For the n-alkanes method, herbage intake was slightly overestimated (7%) when herbage proportion in the diet was high, due to a ratio of faecal C33 : C32 recovery >1. The high bias for the Yb/faecal index was due to the cumulative effect of overestimation of FO (mean recovery of Yb = 0.92) and underestimation of the diet indigestible fraction (-8%). Between-treatment variations of FO were on average well estimated by Yb. Between-treatment variations of OM digestibility estimated using the faecal index technique were lower than those observed in vivo. It is concluded that intake of grazing dairy cows receiving high levels of maize silage supplement should be estimated using the n-alkanes method.     

Effect of offering dairy cows diets differing in phosphorus concentration over four successive lactations: 2. Health, fertility, bone phosphorus reserves and nutrient utilisation

This experiment examined the long-term effects of offering diets containing low levels of dietary phosphorus (P) on dairy cow health, fertility and bone composition, and the effect of dietary P level on nutrient utilisation. One hundred winter-calving Holstein-Friesian dairy cows were offered diets containing either 'high' or 'low' levels of dietary P over a 4-year period. Rations offered during the winter included grass silage, maize silage (70 : 30 dry matter (DM) basis, approximately) and concentrates (10.0 to 12.0 kg/cow per day). During the summer periods in years 1 and 2, half of the cows grazed both day and night, while the remaining cows grazed by day, and were housed by night and offered grass silage. During years 3 and 4, all cows grazed both day and night during the summer period. Concentrate feed levels during the summer periods were 3.0 to 4.0 kg/cow per day. Different dietary P levels were achieved by offering concentrates containing either high or low P levels during the winter period (approximately 7.0 or 4.4 g P/kg DM, respectively) and during the summer period (approximately 6.8 or 3.6 g P/kg DM, respectively). Total ration P levels averaged 4.9 and 3.6 g P/kg DM for the high and low P winter diets, respectively, and 4.2 and 3.6 g P/kg DM for the high and low P summer diets, respectively. A total of 95, 70, 50 and 22 cows completed each of lactations from 1 to 4, respectively. Neither the incidence of lameness or mastitis, or milk somatic cell count, were affected by dietary P level (P > 0.05), while none of the fertility parameters recorded in any of lactations from 1 to 4 was affected by the dietary P level (P > 0.05). Dietary P level had no effect on the specific gravity, ash or calcium content of rib cortical bone cores (n = 78 cows), while the P content of cortical bone (g/kg fresh, g/kg DM and mg/ml fresh bone) was lower with cows offered low P diets (P < 0.05). Dietary P level had no significant effect on the digestibility of either the DM, nitrogen, energy or acid detergent fibre fraction of the diet (P > 0.05), while faecal P excretions were reduced by a mean of 27 g/cow per day with cows offered the low P diets during the winter period. The results of this study indicate that dietary P levels can be reduced to proportionately 0.8 (approximately) of current UK feeding standards (Agricultural and Food Research Council, 1991), with no detrimental effect on dairy cow health or fertility, while having only minor effects on bone composition.     

Effects of faba bean,blue lupin and rapeseed meal supplementation on nitrogen digestion and utilization of dairy cows fed grass silage-based diets

There is increasing interest in using locally produced protein supplements in dairy cow feeding. The objective of this experiment was to compare rapeseed meal (RSM), faba beans (FBs) and blue lupin seeds (BL) at isonitrogenous amounts as supplements of grass silage and cereal based diets. A control diet (CON) without protein supplement was included in the experiment. Four lactating Nordic Red cows were used in a 4 × 4 Latin Square design with four 21 d periods. The milk production increased with protein supplementation but when expressed as energy corrected milk, the response disappeared due to substantially higher milk fat concentration with CON compared to protein supplemented diets. Milk protein output increased by 8.5, 4.4 and 2.7% when RSM, FB and BL were compared to CON. The main changes in rumen fermentation were the higher propionate and lower butyrate proportion of total rumen volatile fatty acids when the protein supplemented diets were compared to CON. Protein supplementation also clearly increased the ruminal ammonia N concentration. Protein supplementation improved diet organic matter and NDF digestibility but efficiency of microbial protein synthesis per kg organic matter truly digested was not affected. Flow of microbial N was greater when FB compared to BL was fed. All protein supplements decreased the efficiency of nitrogen use in milk production. The marginal efficiency (amount of additional feed protein captured in milk protein) was 0.110, 0.062 and 0.045 for RSM, FB and BL, respectively. The current study supports the evidence that RSM is a good protein supplement for dairy cows, and this effect was at least partly mediated by the lower rumen degradability of RSM protein compared to FB and BL. The relatively small production responses to protein supplementation with simultaneous decrease in nitrogen use efficiency in milk production suggest that economic and environmental consequences of protein feeding need to be carefully considered.     

Digestive enzymes and metabolic profile of Labeo rohita fingerlings fed diets with different crude protein levels

Labeo rohita, commonly called rohu is one of the most important fish species for aquaculture in India. Digestive enzyme response and metabolic profile of fingerling L. rohita to different dietary crude protein (CP) levels (viz. 25, 30, 35 and 40%) were studied in an attempt to optimize a practical diet formulation for this species. After 45 days of feeding, activity of digestive enzymes and metabolite concentrations were assayed. Amylase, lipase and alkaline phosphatase (ALP) activities were not influenced by the dietary protein, but proteolytic and acid phosphatase (ACP) activities varied (P<0.05) between the treatments. Proteolytic activity showed a second order polynomial relationship with dietary crude protein (CP) as Y = 0.0734X(2) + 4.937X - 68.37, r(2)=0.97. A positive correlation was observed between dietary CP and amylase (r(2)=0.78). All the metabolites except muscle glucose showed significant change corresponding to the dietary protein levels. Glucose and glycogen levels corresponded to the dietary carbohydrate levels. Muscle and plasma pyruvic acid increased as the crude protein in the diet increased, whereas liver pyruvic acid showed the opposite trend. Muscle protein content was not affected by dietary CP. Protein fractions in plasma (total protein, albumin and globulin) showed maximum values in 30% CP fed group. It is concluded that proteolytic activity and ACP are the major digestive enzymes responsive to dietary CP in L. rohita fingerlings. Considering the cost effectiveness of the diet, and based on liver and plasma free amino acid levels and plasma protein fractions, 30% crude protein is recommended as the optimal dietary protein for L. rohita fingerlings.     

Nitrogen fixation by white clover in pastures grazed by dairy cows: Temporal variation and effects of nitrogen fertilization

Effects of rate of nitrogen (N) fertilizer and stocking rate on production and N₂ fixation by white clover (Trifolium repens L.) grown with perennial ryegrass (Lolium perenne L.) were determined over 5 years in farmlets near Hamilton, New Zealand. Three farmlets carried 3.3 dairy cows ha^–1 and received urea at 0, 200 or 400 kg N ha^–1 yr^–1 in 8–10 split applications. A fourth farmlet received 400 kg N ha^–1 yr^–1 and had 4.4 cows ha^–1.There was large variation in annual clover production and total N₂ fixation, which in the 0 N treatment ranged from 9 to 20% clover content in pasture and from 79 to 212 kg N fixed ha^–1 yr^–1. Despite this variation, total pasture production in the 0 N treatment remained at 75–85% of that in the 400 N treatments in all years, due in part to the moderating effect of carry-over of fixed N between years.Fertilizer N application decreased the average proportion of clover N derived from N₂ fixation (P_N; estimated by ¹⁵N dilution) from 77% in the 0 N treatment to 43–48% in the 400 N treatments. The corresponding average total N₂ fixation decreased from 154 kg N ha^–1 yr^–1 to 39–53 kg N ha^–1 yr^–1. This includes N₂ fixation in clover tissue below grazing height estimated at 70% of N₂ fixation in above grazing height tissue, based on associated measurements, and confirmed by field N balance calculations. Effects of N fertilizer on clover growth and N₂ fixation were greatest in spring and summer. In autumn, the 200 N treatment grew more clover than the 0 N treatment and N₂ fixation was the same. This was attributed to more severe grazing during summer in the 0 N treatment, resulting in higher surface soil temperatures and a deleterious effect on clover stolons.In the 400 N treatments, a 33% increase in cow stocking rate tended to decrease P_N from 48 to 43% due to more N cycling in excreta, but resulted in up to 2-fold more clover dry matter and N₂ fixation because lower pasture mass reduced grass competition, particularly during spring.