Carbohydrate-rich supplements can improve nitrogen use efficiency and mitigate nitrogenous gas emissions from the excreta of dairy cows grazing temperate grass

Temperate pasture species constitute a source of protein for dairy cattle. On the other hand, from an environmental perspective, their high N content can increase N excretion and nitrogenous gas emissions by livestock. This work explores the effect of energy supplementation on N use efficiency (NUE) and nitrogenous gas emissions from the excreta of dairy cows grazing a pasture of oat and ryegrass. The study was divided into two experiments: an evaluation of NUE in grazing dairy cows, and an evaluation of N-NH₃ and N-N₂O volatilizations from dairy cow excreta. In the first experiment, 12 lactating Holstein × Jersey F1 cows were allocated to a double 3 × 3 Latin square (three experimental periods of 17 days each) and subjected to three treatments: cows without supplementation (WS), cows supplemented at 4.2 kg DM of corn silage (CS) per day, and cows supplemented at 3.6 kg DM of ground corn (GC) per day. In the second experiment, samples of excreta were collected from the cows distributed among the treatments. Aliquots of dung and urine of each treatment plus one blank (control – no excreta) were allotted to a randomized block design to evaluate N-NH₃ and N-N₂O volatilization. Measurements were performed until day 25 for N-NH₃ and until day 94 for N-N₂O. Dietary N content in the supplemented cows was reduced by 20% (P < 0.001) compared with WS cows, regardless of the supplement. Corn silage cows had lower N intake (P < 0.001) than WS and GC cows (366 v. 426 g/day, respectively). Ground corn supplementation allowed cows to partition more N towards milk protein compared with the average milk protein of WS cows or those supplemented with corn silage (117 v. 108 g/day, respectively; P < 0.01). Thus, even though they were in different forms, both supplements were able to increase (P < 0.01) NUE from 27% in WS cows to 32% in supplemented cows. Supplementation was also effective in reducing N excretion (761 v. 694 g/kg of N_intake; P < 0.001), N-NH₃ emission (478 v. 374 g/kg of N_milk; P < 0.01) and N-N₂O emission (11 v. 8 g/kg of N_milk; P < 0.001). Corn silage and ground corn can be strategically used as feed supplements to improve NUE, and they have the potential to mitigate N-NH₃ and N-N₂O emissions from the excreta of dairy cows grazing high-protein pastures.     

Beet pulp substituted for corn silage and barley grain in diets fed to dairy cows in the summer months: feed intake,total-tract digestibility,and milk production

The responses of dairy cows to the substitution of beet pulp (BP) for grain or forage are not consistent, and heat stress may affect the response of dairy cows to this substitution. The effects of substituted BP for corn silage and barley grain on feed intake, performance, and ruminal parameters were evaluated using eight multiparous Holstein cows in a duplicated 4 × 4 Latin square design with 21-day periods. Cows were in mid-lactation (45.4 ± 3.6 kg/day milk production and 116 ± 10 days in milk) with an average BW of 664 ± 41.2 kg. Dietary treatments were as follows: 1) 0% BP (0BP, control, 38.5% barley grain, and 20.3% corn silage); 2) 12% BP (12BP, 32.5% barley grain, and 14.3% corn silage); 3) 18% BP (18BP, 29.5% barley grain, and 11.3% corn silage); and 4) 24% BP (24BP, 26.5% barley grain, and 8.3% corn silage). Cows were under mild heat stress and the average temperature–humidity index was 70.5; increasing BP caused a linear decrease in respiration rate (P < 0.01). Higher BP in the diet caused a linear increase in DM intake (P = 0.01) and NDF digestibility (P = 0.03). Dry and organic matter (OM) digestibilities tended to increase linearly with higher BP (P < 0.10). Milk yield, energy-corrected milk, protein, lactose, and fat production and content were not affected by the treatments. Increasing BP in the diet caused a linear decrease in feed efficiency and rumen ammonia (P < 0.05) and a tendency to a linear decrease in milk urea nitrogen (P < 0.10). Rumen pH and acetate to propionate ratio were not affected by the replacement. Total volatile fatty acid concentration in the rumen increased linearly with increasing the BP inclusion (P = 0.04). Acetate and butyrate (P = 0.07) proportion tended to increase, whereas propionate (P = 0.06) and isovalerate (P = 0.08) proportion tended to decrease linearly as BP was substituted for corn silage and barley grain. The results indicated that under mild heat stress condition, BP can be successfully substituted for barley grain and corn silage up to 24% of the diet without any negative effect on production and ruminal pH.     

Nitrogen isotopic fractionation as a biomarker for nitrogen use efficiency in ruminants: a meta-analysis

Animal proteins are naturally ¹⁵N enriched relative to the diet and the extent of this difference (Δ¹⁵N_animal-diet or N isotopic fractionation) has been correlated to N use efficiency (NUE; N gain or milk N yield/N intake) in some recent ruminant studies. The present study used meta-analysis to investigate whether Δ¹⁵N_animal-diet can be used as a predictor of NUE across a range of dietary conditions, particularly at the level of between-animal variation. An additional objective was to identify variables related to N partitioning explaining the link between NUE and Δ¹⁵N_animal-diet. Individual values from eight publications reporting both NUE and Δ¹⁵N_animal-diet for domestic ruminants were used to create a database comprising 11 experimental studies, 41 treatments and individual animal values for NUE (n=226) and Δ¹⁵N_animal-diet (n=291). Data were analyzed by mixed-effect regression analysis taking into account experimental factors as random effects on both the intercept and slope of the model. Diets were characterized according to the INRA feeding system in terms of N utilization at the rumen, digestive and metabolic levels. These variables were used in a partial least squares regression analysis to predict separately NUE and Δ¹⁵N_animal-diet variation, with the objective of identifying common variables linking NUE and Δ¹⁵N_animal-diet. For individuals reared under similar conditions (within-study) and at the same time (within-period), the variance of NUE and Δ¹⁵N_animal-diet not explained by dietary treatments (i.e. between-animal variation plus experimental error) was 35% and 55%, respectively. Mixed-effect regression analysis conducted with treatment means showed that Δ¹⁵N_animal-diet was significantly and negatively correlated to NUE variation across diets (NUE=0.415 −0.055×Δ¹⁵N_animal-diet). When using individual values and taking into account the random effects of study, period and diet, the relationship was also significant (NUE=0.358 −0.035×Δ¹⁵N_animal-diet). However, there may be a biased prediction for animals close to zero, or in negative, N balance. When using a novel statistical approach, attempting to regress between-animal variation in NUE on between-animal variation in Δ¹⁵N_animal-diet (without the influence of experimental factors), the negative relationship was still significant, highlighting the ability of Δ¹⁵N_animal-diet to capture individual variability. Among the studied variables related to N utilization, those concerning N efficiency use at the metabolic level contributed most to predict both Δ¹⁵N_animal-diet and NUE variation, with rumen fermentation and digestion contributing to a lesser extent. This study confirmed that on average Δ¹⁵N_animal-diet can predict NUE variation across diets and across individuals reared under similar conditions.     

Relationship between efficiency of nitrogen utilization and isotopic nitrogen fractionation in dairy cows: contribution of digestion v. metabolism?

Animal tissues are naturally ¹⁵N enriched relative to their diet and the extent of this difference (Δ¹⁵N_animal-diet) has been correlated to the efficiency of N assimilation in different species. The rationale is that transamination and deamination enzymes, involved in amino acid metabolism are likely to preferentially convert amino groups containing ¹⁴N over ¹⁵N. However, in ruminants the contribution of rumen bacterial metabolism relative to animal tissues metabolism to naturally enrich animal proteins in terms of ¹⁵N has been not assessed yet. The objective of this study was to assess the impact of rumen and digestion processes on the relationship between Δ¹⁵N_animal-diet and efficiency of N utilization for milk protein yield (milk N efficiency (MNE); milk N yield/N intake) as well as the relationship between the ¹⁵N natural abundance of rumen bacteria and the efficiency of N use at the rumen level. Solid- and liquid-associated rumen bacteria, duodenal digesta, feces and plasma proteins were obtained (n=16) from four lactating Holstein cows fed four different diets formulated at two metabolizable protein supplies (80% v. 110% of protein requirements) crossed by two different dietary energy source (diets rich in starch v. fiber). We measured the isotopic N fractionation between animal and diet (Δ¹⁵N_animal-diet) in these different body pools. The Δ¹⁵N_animal-diet was negatively correlated with MNE when measured in solid-associated rumen bacteria, duodenal digesta, feces and plasma proteins, with the strongest correlation found for the latter. However, our results showed a very weak ¹⁵N enrichment of duodenal digesta (Δ¹⁵N_{duodenal digesta-diet} mean value=0.42) compared with that observed in plasma proteins (Δ¹⁵N_{plasma protein-diet} mean value=2.41). These data support the idea that most of the isotopic N fractionation observed in ruminant proteins (Δ¹⁵N_{plasma protein-diet}) has a metabolic origin with very little direct impact of the overall digestion process on the existing relationship between Δ¹⁵N_{plasma protein-diet} and MNE. The ¹⁵N natural abundance of rumen bacteria was not related to either rumen N efficiency (microbial N/available N) or digestive N efficiency (metabolizable protein supply/CP intake), but showing a modest positive correlation with rumen ammonia concentration. When using diets not exceeding recommended protein levels, the contribution of rumen bacteria and digestion to the isotopic N fractionation between animal proteins and diet is low. In our conditions, most of the isotopic N fractionation (Δ¹⁵N_{plasma protein-diet}) could have a metabolic origin, but more studies are warranted to confirm this point with different diets and approaches.     

Nitrogen partitioning and isotopic discrimination are affected by age and dietary protein content in growing lambs

It is difficult to separate an age-dependent fall in nitrogen use efficiency (NUE; N balance/N intake) in growing ruminants from a progressively decrease in animal protein requirements over time. This study examined the effect of dietary protein content on N partitioning, digestibility and N isotopic discrimination between the animal and its diet (Δ¹⁵N_animal-diet) evaluated at two different fattening periods (early v. late). Twenty-four male Romane lambs (age: 19 ± 4.0 days; BW: 8.3 ± 1.39 kg) were equally allocated to three dietary CP treatments (15%, 17% and 20% CP on a DM basis). Lambs were reared with their mothers until weaning, thereafter housed in individual pens until slaughter (45 kg BW). During the post-weaning period, lambs were allocated twice (early fattening (30 days post-weaning) and late fattening (60 days post-weaning)) to metabolic cages for digestibility and N balance study. When diet CP content increased, the average daily gain of lambs increased (P < 0.05) while the age at slaughter decreased (P = 0.01), but no effect was observed on feed efficiency (P > 0.10). Diet CP content had limited effect on lamb carcass traits. Higher fibre digestibility was observed at the early v. late fattening period (P < 0.001). The N intake and the urinary N excretion increased when diet CP content increased (P < 0.001) and when shifting from early to late fattening period (P < 0.001). Faecal N excretion (P = 0.14) and N balance (P > 0.10) were not affected by diet CP content. Nitrogen digestibility increased (P < 0.001) as the diet CP content increased and on average it was greater at late v. early fattening period (P = 0.02). The NUE decreased (P = 0.001) as the diet CP content increased and as the lamb became older (P < 0.001). However, the age-dependent fall in NUE observed was lower at high v. low dietary CP content (CP × age interaction; P = 0.04). The Δ¹⁵N_animal-diet was positively correlated (P < 0.05) with N intake (r = 0.59), excretion of faecal N (r = 0.41), urinary N (r = 0.69) and total manure N (r = 0.64), while negatively correlated with NUE (r = −0.57). Overall, the experiment showed NUE was lower in older lambs and when lambs were fed high diet CP content, and that Δ¹⁵N_animal-diet was a useful indicator not only for NUE but also for urinary N excretion, which is a major environmental pollution factor on farm.     

Milk production responses and rumen fermentation of dairy cows supplemented with summer brassicas

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.     

Long-term effect of linseed plus nitrate fed to dairy cows on enteric methane emission and nitrate and nitrite residuals in milk

A previous study showed the additive methane (CH₄)-mitigating effect of nitrate and linseed fed to non-lactating cows. Before practical application, the use of this new strategy in dairy cows requires further investigation in terms of persistency of methanogenesis reduction and absence of residuals in milk products. The objective of this experiment was to study the long-term effect of linseed plus nitrate on enteric CH₄ emission and performance in dairy cows. We also assessed the effect of this feeding strategy on the presence of nitrate residuals in milk products, total tract digestibility, nitrogen (N) balance and rumen fermentation. A total of 16 lactating Holstein cows were allocated to two groups in a randomised design conducted in parallel for 17 weeks. Diets were on a dry matter (DM) basis: (1) control (54% maize silage, 6% hay and 40% concentrate; CON) or (2) control plus 3.5% added fat from linseed and 1.8% nitrate (LIN+NIT). Diets were equivalent in terms of CP (16%), starch (28%) and NDF (33%), and were offered twice daily. Cows were fed ad libitum, except during weeks 5, 16 and 17 in which feed was restricted to 95% of dry matter intake (DMI) to ensure complete consumption of meals during measurement periods. Milk production and DMI were measured weekly. Nitrate and nitrite concentrations in milk and milk products were determined monthly. Daily CH₄ emission was quantified in open circuit respiration chambers (weeks 5 and 16). Total tract apparent digestibility, N balance and rumen fermentation parameters were determined in week 17. Daily DMI tended to be lower with LIN+NIT from week 4 to 16 (−5.1 kg/day on average). The LIN+NIT diet decreased milk production during 6 non-consecutive weeks (−2.5 kg/day on average). Nitrate or nitrite residuals were not detected in milk and associated products. The LIN+NIT diet reduced CH₄ emission to a similar extent at the beginning and end of the trial (−47%, g/day; −30%, g/kg DMI; −33%, g/kg fat- and protein-corrected milk, on average). Diets did not affect N efficiency and nutrients digestibility. In the rumen, LIN+NIT did not affect protozoa number but reduced total volatile fatty acid (−12%) and propionate (−31%) concentrations. We concluded that linseed plus nitrate may have a long-term CH₄-mitigating effect in dairy cows and that consuming milk products from cows fed nitrate may be safe in terms of nitrate and nitrite residuals. Further work is required to optimise the doses of linseed plus nitrate to avoid reduced cows performance.     

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.     

Effect of substituting wet corn gluten feed and corn stover for alfalfa hay in total mixed ration silage on lactation performance in dairy cows

Wet corn gluten feed (WCGF) is a high moisture feed containing rapidly digestible, non-forage fiber and protein. The objective of this study was to investigate the effect of substituting WCGF and corn stover for alfalfa hay in total mixed ration (TMR) silage on lactation performance and nitrogen balance in dairy cows. Nine multiparous Holstein dairy cows (BW = 532 ± 28.9 kg and day in milk = 136 ± 5.6 d; mean ± SD) were used in a replicated 3 × 3 Latin square design with 21-d periods (14 d of diet adaption and 7 d of sample collection). Groups were balanced for parity, day in milk, and milk production and consumed one of three treatment diets during each period. The treatment diets were fed as TMR and contained similar concentrate mixtures and corn silage but different proportions of roughage and WCGF. The three treatments were: (1) 0% WCGF, 0% corn stover, and 22.1% alfalfa hay (0% WCGF); (2) 6.9% WCGF, 3.4% corn stover, and 11.8% alfalfa hay (7% WCGF); and (3) 13.3% WCGF, 4.9% corn stover, and 3.9% alfalfa hay (13.3% WCGF). Compared to the 0% WCGF diet, the cows fed the 7% and 13.3% WCGF diets had a higher milk yield and concentration of milk fat, protein, lactose, and total solids. Effective degradability of DM was higher in the cows fed the 7% and 13.3% WCGF diets than it was with the 0% WCGF diet. Cows fed the 13.3% WCGF had a higher CP effective degradability and a lower rumen undegraded protein than cows fed the 0% WCGF diet. The concentration of ruminal volatile fatty acids and ammonia-N was higher in cows fed the 7% and 13.3% WCGF diets than cows fed the 0% WCGF diet. The fecal N was lower in cows fed the 7% and 13.3% WCGF diets than it was in cows fed the 0% WCGF diet. Milk N secretion and milk N as a percent of N intake were higher in cows fed the 13.3% WCGF diet than cows fed the 0% and 7% WCGF diets. In conclusion, it appears that feeding a TMR silage containing WCGF and corn stover in combination, replacing a portion of alfalfa hay, may improve lactation performance and nitrogen utilization for lactating dairy cows.     

Effects of replacing soybean meal with pumpkin seed cake and dried distillers grains with solubles on milk performance and antioxidant functions in dairy cows

Pumpkin seed cake (PSC), a byproduct of pumpkin seed oil processing, is used in ruminant feed as a beneficial protein source. Experiments were conducted to evaluate PSC as a substitute for soybean meal in the diets of lactating cows based on performance, rumen fermentation, antioxidant function and nitrogen partitioning. Six multiparous lactating cows were used in a replicated 3 × 3 Latin square experiment with 27-day periods. The cows were randomly divided into three treatment groups: group (1) was fed a diet containing no PSC (0PSC), and groups (2) and (3) were fed diets in which soybean meal was replaced with PSC and dried distillers grains with solubles (DDGS) at levels of 50% (50PSC) and 100% (100PSC), respectively. The diets were isonitrogenous and contained identical roughage but different proportions of PSC and DDGS. Replacement of soybean meal with PSC and DDGS did not influence rumen degradation, milk performance, rumen fermentation, DM intake or apparent total tract digestibility, and nitrogen partitioning between milk, feces and urine did not differ in the animals fed the three diets. However, compared with a diet containing no PSC, the total antioxidant capacity (P < 0.05) and antioxidant enzymes (total superoxide dismutase, glutathione peroxidase and catalase) activities (P < 0.05) were increased in the animals that received the 50PSC and 100PSC diets. In contrast, addition of PSC significantly reduced concentrations of aspartate transaminase (P < 0.05), alkaline phosphatase (P < 0.05) and malondialdehyde (P < 0.05) in the plasma. These results demonstrate that PSC can be completely substituted for soybean meal in the diet of dairy cows without any negative impact on milk performance, rumen fermentation or apparent digestibility and that this dietary change improves antioxidant functions and blood parameters in dairy cows, indicating that PSC has the potential for use as a feed source for dairy cows.     

Effects of condensed tannin-amended cassava silage blend diets on feeding behavior,digestibility, nitrogen balance,milk yield and milk composition in dairy goats

Condensed tannins (CTs) are phenolic compounds derived from secondary plant metabolism that act as part of the plant's chemical defense system against pathogen invasion and herbivorous attack. This study aimed to evaluate the intake, digestibility, nitrogen (N) balance, production and composition of milk from goats fed cassava silage with added levels of CTs. Eight Anglo-Nubian goats with a mean BW of 40 ± 2.0 kg were distributed in a double Latin square design with four levels of CTs (0, 25, 50 and 75 g/kg DM) with four 20-day periods with 15 days of adaptation and five evaluation days for each period. No differences were observed in DM, NDF, CP intake and feed conversion (grams of DM intake (DMI) per gram of milk produced); however, when expressed as percent of BW, DMI showed a quadratic increase to 29.1 g/kg. As the level of supplemented CTs increased in the diet, the CP digestibility (P = 0.023), NDF (P = 0.044), non-fiber carbohydrates (NFC; P = 0.032) and total digestible nutrients (P = 0.033) exhibited a linear decrease. Furthermore, the addition of CTs to cassava silage induced a linear increase in N-fecal excretion (P = 0.014) and a positive quadratic effect on N-retained (P = 0.014) and N-balance (P = 0.024) as well as a positive quadratic trend in N-digested (P = 0.092). Milk urea N (P = 0.023) decreased linearly. The addition of CTs to cassava silage had a positive quadratic effect on ruminating time (P = 0.011). In addition, comparing the use or non-use from the orthogonal contrast test, the inclusion of CTs in goat diet increased water and N-intake, CP and NDF digestibility, spent time eating and ruminating and N-balance and decreased milk production corrected^3.5%, fat milk content, milk urea N and dry defatted extract of milk. Thus, adding CTs to cassava silage at 25 g/kg total DM promoted goats' greater use of the diet without impairing feed conversion and the quality of goat milk produced. Dietary levels of 50 and 75 g/kg total DM are not recommended because under the conditions of this study, they reduced the productive efficiency of dairy goats.     

Grass silage pulp as a dietary component for high-yielding dairy cows

Green biorefineries provide novel opportunities to use the green biomass efficiently and utilize the ecosystem services provided by grasslands more widely. The effects of the inclusion of fractionated grass silage solid fraction (pulp) on feed intake, rumen fermentation, diet digestion and milk production in dairy cows were investigated. Pulp was separated from grass silage using a screw press simulating a green biorefinery. Partial removal of liquid from forage increased DM concentration from 220 to 432 g/kg and NDF from 589 to 709 g/kg DM while CP decreased from 144 to 107 g/kg DM. A feeding trial using an incomplete changeover design with 24 Nordic Red cows and two 3-week periods was conducted. The pulp replaced grass silage in the diet at 0 (P0), 25 (P25) and 50 (P50) percentage of total forage, which was fed ad libitum with 13 kg of concentrate for all treatments. The forage DM intake was highest on P25 (14.1 kg/day) while P0 and P50 did not differ from each other (13.2 and 13.0 kg/day, respectively). There were no differences between the treatments in rumen pH or ammonia N, but the proportion of acetate increased with increasing pulp inclusion. The digestibility was measured using acid insoluble ash and indigestible NDF (iNDF) as internal markers. Neither of the markers detected differences in NDF digestibility, but according to iNDF, apparent total tract organic matter digestibility decreased with increasing pulp inclusion. The cows maintained milk production at P25, but it showed some decline at P50 (energy-corrected milk at P0 and P25 was 39.8 kg/day while for P50, it was 38.5 kg/day, P = 0.056) and the milk protein yield significantly declined with higher pulp inclusion. Simultaneously, the nitrogen use efficiency in milk production increased. It seems that the fibrous grass-based fraction from a biorefinery process has potential to be used as a feed for ruminants.     

Effects of dehydrated lucerne and soya bean meal on milk production and composition,nutrient digestion,and methane and nitrogen losses in dairy cows receiving two different forages

Dehydrated lucerne is used as a protein source in dairy cow rations, but little is known about the effects of lucerne on greenhouse gas production by animals. Eight Holstein dairy cows (average weight: 582 kg) were used in a replicated 4×4 Latin square design. They received diets based on either maize silage (M) or grass silage (G) (45% of diet on dry matter (DM) basis), with either soya bean meal (15% of diet DM) completed with beet pulp (15% of diet DM) (SP) or dehydrated lucerne (L) (30% of diet DM) as protein sources; MSP, ML, GSP and GL diets were calculated to meet energy requirements for milk production by dairy cows and degradable protein for rumen microbes. Dry matter intake (DMI) did not differ among diets (18.0 kg/day DMI); milk production was higher with SP diets than with L diets (26.0 v. 24.1 kg/day), but milk production did not vary with forage type. Milk fatty-acid (FA) composition was modified by both forage and protein sources: L and G diets resulted in less saturated FA, less linoleic acid, more trans-monounsaturated FA, and more linolenic acid than SP and M diets, respectively. Enteric methane (CH₄) production, measured by the SF₆ tracer method, was higher for G diets than for M diets, but did not differ with protein source. The same effects were observed when CH₄ was expressed per kg milk. Minor effects of diets on rumen fermentation pattern were observed. Manure CH₄ emissions estimated from faecal organic matter were negatively related to diet digestibility and were thus higher for L than SP diets, and higher for M than G diets; the resulting difference in total CH₄ production was small. Owing to diet formulation constraints, N intake was higher for SP than for L diets; interaction between forage type and protein source was significant for N intake. The same statistical effects were found for N in milk. Faecal and urinary N losses were determined from total faeces and urine collection. Faecal N output was lower for M than for G diets but did not differ between protein sources. Urinary N output did not differ between forage types, but was lower for cows fed L diets than for cows fed SP diets, potentially resulting in lower ammonia emissions with L diets. Replacing soya bean meal plus beet pulp with dehydrated lucerne did not change CH₄ production, but resulted in more N in faeces and less N in urine.     

Effects of a combination of fibrolytic and amylolytic enzymes on ruminal enzyme activities,bacterial diversity,blood profile and milk production in dairy cows

We hypothesised that adding a combination of fibrolytic and amylolytic enzymes to the diet of early-lactation dairy cows would improve rumen enzyme activity and bacterial diversity, promote energy metabolism, and benefit milk production in cows. Twenty multiparous early-lactation (90 ± 5 d) Holstein cows with similar body conditions were randomly allocated to control (CON, n = 10) and experimental (EXP, n = 10) groups in a completely randomised single-factor design. The CON was fed only a basal total mixed ration diet, and the diet of the EXP was supplemented with a combination of fibrolytic and amylolytic enzymes at 70 g/cow/d (cellulase 3 500 CU/g, xylanase 2 000 XU/g, β-glucanase 17 500 GU/g, and amylase 37 000 AU/g). The experiment lasted 28 days, with 21 days for adaptation and 7 days for sampling. Enzyme addition increased the activity levels of α-amylase and xylanase, and the ammonia-N concentration (P < 0.05) tended to increase the activity of β-glucanase (P = 0.08) in rumen fluid. However, there was no significant difference in the rumen bacterial richness and diversity, phylum (richness > 0.1%) or genus (richness > 1%) composition between the CON and EXP groups (P > 0.05). A tendency of difference was found between CON and EXP (R = 0.22, P = 0.098) in principal component analysis. Ten genera showed different abundances across the CON and EXP groups (linear discriminant analysis effect size, linear discriminant analysis > 2). EXP increased the ratio of albumin to globulin and the concentrations of total cholesterol and low-density lipoprotein cholesterol (P < 0.05) and tended to increase triglycerides (P = 0.09) in blood. Milk yield, 3.5% fat-corrected milk yield and energy-corrected milk yield increased with enzyme supplementation (P < 0.05). The production levels of milk fat and lactose increased, but the percentage of solids, not fat and protein, decreased in EXP (P < 0.05). Although the DM intake was not affected, the feed efficiency tended to increase (P = 0.07) in EXP. In conclusion, dietary supplementation with a mixture of fibrolytic and amylolytic enzymes on multiparous early-lactation dairy cows increased α-amylase and xylanase activity levels in rumen fluid, enhanced milk performance and tended to improve the feed efficiency in cows.     

Lactational performance,feeding behavior,ruminal fermentation and nutrient digestibility in dairy cows fed whole-plant faba bean silage-based diet with fibrolytic enzyme

Whole-plant faba bean silage has a high content in indigestible fiber. Improvement of fiber digestibility of faba bean silage would benefit animal production. However, there is no study on pretreating fibrolytic enzyme in whole-plant faba bean silage-based diet for dairy cows on animal performance. The objectives of this study were to evaluate the effects of pretreating whole-plant faba bean silage-baseddiet with fibrolytic enzyme (a mixture of xylanase and cellulase; AB Vista, UK) derived from Trichoderma reesei (FETR) on lactational performance, digestibility, ruminal fermentation characteristics, and feeding behavior of dairy cows. The animal trial was conducted using eight lactating Holstein cows (BW = 710 ± 44 kg and Days in Milk (DIM) = 121 ± 17 days) with four levels of FETR (0, 0.5, 0.75, and 1.0 mL of FETR/kg DM of silage) in a replicated Latin square design. These enzyme treatments were selected based on the previous in situ and in vitro findings that showed positive responses to the whole-plant faba bean silage. The enzyme treatments were directly applied on the silage prior to mixing process. The total mixed rations contained 31% of faba bean silage, 14% of grass hay, 3.5% of straw, 30% of barley and corn grain and 21.5% of concentrate. There was no significant difference of applying FETR on nutrient intake (P > 0.05) except for CP intake, which was reduced in FETR group compared to control (P < 0.01, 4.4 vs 4.54 kg/d). There was a linear effect found in NDF digestibility when treated with FETR, where maximum improvement was achieved with 0.5 mL of FETR application. The milk fat yield, percentage of milk fat and fat-corrected milk were linearly affected by the increasing level of enzyme. The cows fed a diet supplemented with enzymes tended to have a lower milk fat. Feed efficiency linearly responded to incremental levels of FETR. There was no enzyme effect on feeding behavior and nitrogen balance and utilization. Results from this study indicated that supplementing fibrolytic enzyme on whole-plant faba bean silage diets for dairy cows improved lactational performance, intake and digestibility with 0.5 mL of FETR application. However, adding higher enzyme level resulted in negative effects on animal performance.     

Rubber seed oil and flaxseed oil supplementation alter digestion,ruminal fermentation and rumen fatty acid profile of dairy cows

Rubber seed oil (RO) that is rich in polyunsaturated fatty acids (FA) can improve milk production and milk FA profiles of dairy cows; however, the responses of digestion and ruminal fermentation to RO supplementation in vivo are still unknown. This experiment was conducted to investigate the effect of RO and flaxseed oil (FO) supplementation on nutrients digestibility, rumen fermentation parameters and rumen FA profile of dairy cows. Forty-eight mid-lactation Holstein dairy cows were randomly assigned to one of four treatments for 8 weeks, including basal diet (CON) or the basal dietary supplemented with 4% RO, 4% FO or 2% RO plus 2% FO on a DM basis. Compared with CON, dietary oil supplementation improved the total tract apparent digestibility of DM, neutral detergent fibre and ether extracts ( P < 0.05). Oil treatment groups had no effects on ruminal digesta pH value, ammonia N and microbial crude protein ( P > 0.05), whereas oil groups significantly changed the volatile fatty acid (VFA) profile by increasing the proportion of propionate whilst decreasing total VFA concentration, the proportion of acetate and the ratio of acetate to propionate ( P < 0.05). However, there were no differences in VFA proportions between the three oil groups (P > 0.05). In addition, dietary oil supplementation increased the total unsaturated FA proportion in the rumen by enhancing the proportion of trans-11 C18:1 vaccenic acid (VA), cis-9, trans-11 conjugated linoleic acid (CLA) and α-linolenic acid (ALA) ( P < 0.05). These results indicate that dietary supplementation with RO and FO could improve nutrients digestibility, ruminal fermentation and ruminal FA profile by enhancing the VA, cis-9, trans-11 CLA and ALA composition of lactating dairy cows. These findings provide a theoretical basis for the application of RO in livestock production.     

Effects of feeding propylene glycol to mid-lactating dairy cows

Propylene glycol (PG) is a gluconeogenic precursor widely used to prevent and treat ketosis postpartum. The study has investigated the effects of PG administration to dairy cows at mid stage of lactation. According to a 3 × 3 latin square design, three Italian Brown lactating cows (125 ± 7 days in milk) fitted with rumen cannula were fed a corn silage based diet (CP 14.7%, NDF 41.1% DM) with 0, 200 or 400 g day⁻¹ PG added. Dry matter intake was increased by feeding PG with a significant quadratic component per dose (16.2, 17.2 and 16.5 kg day⁻¹ for 0, 200 and 400 g PG day⁻¹, respectively). Milk yield was not affected by PG, averaging 17.1 kg day⁻¹. Average daily gain increased from 64 to 206 and 302 g day⁻¹ when cows received 200 and 400 g day⁻¹ of PG (linear component per dose P < 0.05). Digestibility of the diet did not differ among treatments, whereas repeated rumen fluid samples, taken 0, 2, 4, 6 and 8 h after the meal, showed a consistently lower acetate to propionate ratio when feeding PG. Blood insulin was not affected by PG administration. Except for therapeutic treatments, PG administration to dairy cows at mid stage of lactation should be advised against. Despite the positive effect on intake, administration of the additive, increasing the molar percentage of rumen propionate, contributes to shift the energy partition from milk production to liveweight gain.     

Herbage utilisation method affects rumen fluid and milk fatty acid profile in Holstein and Montbéliarde cows

Compared with maize silage- and concentrate-based diets, herbage-based diets were repeatedly shown to favourably influence the milk fatty acid (FA) profile. However, it is unclear how the herbage feeding mode (grazing vs indoor green-feeding) and conservation (fresh herbage vs hay vs silage) modify the milk FA profile. Therefore, the aim of the present experiment was to investigate the effect of different herbage utilisation methods (including herbage feeding mode and herbage conservation method) on the ruminal biohydrogenation of dietary FA and the consequences on the milk FA composition in cows of two breeds (Holstein and Montbéliarde). Concomitant effects of botanical composition and phenological stage of the herbage on milk FA profile were controlled for by harvesting barn-dried hay and silage simultaneously as first cut from the same ryegrass-dominated grassland in a semi-mountainous region. Seven weeks later, the first regrowth of the same plot was used as fresh herbage, either grazed or fed indoor (indoor green-feeding). Twenty-four Montbéliarde and 24 Holstein cows were randomly allocated to four groups of 12 cows balanced by breed, parity, and milk yield. In a free-stall barn, three groups were given ad libitum access to hay, silage, or fresh herbage, respectively. The fourth group was strip-grazing. All cows were supplemented with 3 kg DM/day of the same energy-rich concentrate. After 2 weeks of adaptation to the forage, samples of forage, concentrate, milk, blood, and rumen fluid were collected. Fatty acid composition of forages, rumen fluid, and milk was analysed by gas chromatography. Haymaking reduced total FA content of the herbage, in particular that of linoleic acid (LA) and α-linolenic acid (ALA). Still, rumen fluid lipids of hay-fed cows had the highest proportion of rumenic acid, LA, ALA, and total polyunsaturated fatty acids (PUFAs). Milk fat from hay-fed cows had the highest proportion of LA, and the apparent transfer rates from feed to milk of LA and ALA were higher in hay-fed cows than in silage-fed cows. The proportion of PUFAs was highest in milk fat from grazing and indoor green-fed Montbéliarde cows and lowest in silage-fed cows of both breeds. In conclusion, the herbage utilisation method affects the ruminal biohydrogenation of LA and ALA, whereby herbage drying particularly increases their transfer from herbage to milk.     

Wet brewers' grains or bean curd pomance as partial replacement of soybean meal for lactating cows

The purpose of this study was to evaluate the effects of supplementation of the by-product, brewers' grains and the bean curd pomance, on the performance of lactating cows and their ruminal characteristics. Through this, we wanted to increase resource utilization and to eliminate pollution from these by-products. Thirty-two Holstein lactating cows were allocated randomly into four dietary treatment groups. The experimental diets were formulated to be isoenergetic and isonitrogenous according to National Research Council (1989), and containing 35% corn silage, 20% alfalfa hay and 45% concentrates on DM basis. The dietary treatments consisted of the inclusion of different by-products sources. These included soybean bean (as the control), brewers' grains, bean curd pomance, and the mixed by-products (containing the same amount of brewers' grains and bean curd pomance). The experimental feeding lasted for eight weeks after one week of adaptation. In addition, four rumen cannulated Holstein cows were used in a 4×4 Latin square with 10-day period for collecting rumen samples. Results showed that cows that were fed the bean curd pomance diet produced significantly more milk than those that were fed the brewers' grains diet or the mixed by-product diet. The cows on the control diet produced significantly less milk than the other treatment groups (P<0.05). Both the control and brewers' grains groups consumed more feed than those in the bean curd pomance and mixed by-products diet groups (P<0.05). The cows that were fed the mixed by-products diet produced a significantly lower percentage of milk fat, total solids, milk protein and milk lactose than the others (P<0.05). The milk lactose percentage was significantly higher for cows that were fed the bean curd pomance than those fed the brewers' grains diet (P<0.05). Cows that were fed the bean curd pomance produced significantly more milk fat, total solids, milk protein and milk lactose than the others (P<0.05). Cows that were fed the mixed by-products diet produced significantly lower amount of milk fat, total solids and milk protein than the control and brewers' grains group (P<0.05). These diets did not significantly influence the body weight of the cows. It, however, significantly influenced the ruminal characteristics (P<0.05).     

Reducing protein content in the diet of growing goats: implications for nitrogen balance,intestinal nutrient digestion and absorption,and rumen microbiota

Reducing dietary CP content is an effective approach to reduce animal nitrogen excretion and save protein feed resources. However, it is not clear how reducing dietary CP content affects the nutrient digestion and absorption in the gut of ruminants, therefore it is difficult to accurately determine how much reduction in dietary CP content is appropriate. This study was conducted to investigate the effects of reduced dietary CP content on N balance, intestinal nutrient digestion and absorption, and rumen microbiota in growing goats. To determine N balance, 18 growing wether goats (25.0 ± 0.5 kg) were randomly assigned to one of three diets: 13.0% (control), 11.5% and 10.0% CP. Another 18 growing wether goats (25.0 ± 0.5 kg) were surgically fitted with ruminal, proximate duodenal, and terminal ileal fistulae and were randomly assigned to one of the three diets to investigate intestinal amino acid (AA) absorption and rumen microbiota. The results showed that fecal and urinary N excretion of goats fed diets containing 11.5% and 10.0% CP were lower than those of goats fed the control diet (P < 0.05). When compared with goats fed the control diet, N retention was decreased and apparent N digestibility in the entire gastrointestinal tract was increased in goats fed the 10% CP diet (P < 0.05). When compared with goats fed the control diet, the duodenal flow of lysine, tryptophan and phenylalanine was decreased in goats fed the 11.5% CP diet (P < 0.05) and that of lysine, methionine, tryptophan, phenylalanine, leucine, glutamic acid, tyrosine, essential AAs (EAAs) and total AAs (TAAs) was decreased in goats fed the 10.0% CP diet (P < 0.05). When compared with goats fed the control diet, the apparent absorption of TAAs in the small intestine was increased in goats fed the 11.5% CP diet (P < 0.05) and that of isoleucine, serine, cysteine, EAAs, non-essential AAs, and TAAs in the small intestine was increased in goats fed the 10.0% CP diet (P < 0.05). When compared with goats fed the control diet, the relative richness of Bacteroidetes and Fibrobacteres was increased and that of Proteobacteria and Synergistetes was decreased in the rumen of goats fed a diet with 10.0% CP. In conclusion, reducing dietary CP content reduced N excretion and increased nutrient utilization by improving rumen fermentation, enhancing nutrient digestion and absorption, and altering rumen microbiota in growing goats.