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Identification of plasma biomarkers for feed efficiency in growing beef cattle offers a promising opportunity for developing prediction models to improve precision feeding strategies. However, these models must accurately predict feed efficiency at early stages of fattening. Our study aimed to evaluate the reliability of candidate biomarkers previously identified in late-fattening cattle when analysed during early fattening stages and to develop diet-specific prediction equations for residual feed intake (RFI). From a total of 364 Charolais bulls across seven cohorts, we selected 64 animals with extreme RFI values. The animals were fed either a corn‑ or grass-silage diets. These animals were chosen from four out of the available seven cohorts. Animals from three cohorts (24 high-RFI and 24 low-RFI, having a mean RFI difference of 1.48 kg/d) were used for biomarker confirmation and prediction model training. Animals from a fourth cohort (8 high-RFI and 8 low-RFI, having a mean RFI difference of 0.98 kg/d) were used for model external validation. Blood samples were collected at the beginning of the feed efficiency test (333 ± 20 days), and plasma underwent targeted metabolomic for 630 metabolites, natural abundance of 15N (δ15N), insulin, and IGF-1 analysis. Seven previously identified plasma biomarkers for RFI in late-fattening beef cattle still kept their capability for discriminating low and high RFI animals when analysed during early fattening stages (P 0.05). Prediction models were developed based on seven confirmed RFI biomarkers analysed during early-fattening. Two logistic regression models incorporating creatinine and either IGF-1 (for grass silage-based diet) or PC aa C30:2 (for corn silage-based diet) effectively distinguished between high− and low-RFI animals with high sensitivity and specificity (area under the curve > 0.80). The biomarkers used in the models showed moderate to high repeatability between early and late fattening stages (0.45

Abstract This study investigated whether plasma biomarkers of residual feed intake (RFI), identified under ad libitum feeding conditions in beef cattle, remained consistent during feed restriction. Sixty Charolais crossbred young bulls were divided into two groups for a crossover study. Group A was initially fed ad libitum (first test) and then restricted (second test) on the same diet, while Group B experienced the opposite sequence. Blood samples were collected from the 12 most divergent RFI animals in each group at the end of the first test and again after the second test. 12 plasma variables consistently increased, while three consistently decreased during feed restriction (FDR

ABSTRACT: The extrusion of leguminous seeds induces the formation of Maillard reaction compounds (MRC) as a product of protein advanced glycation and oxidation, which lowers protein degradability in the rumen. However, the quantitative relationship between the parameters of pretreatment (i.e., addition of reducing sugars) and extrusion, and the formation of MRC has not been established yet. Moreover, the fate of the main stable MRC, Nε-carboxymethyl-lysine (CML), in the excretory routes has never been investigated in ruminants. We aimed to test the effects of the temperature of extrusion of white lupines with or without addition of reducing sugars on the formation of MRC, crude protein (CP) degradability in the rumen, N use efficiency for milk production (milk N/N intake), and performance of dairy cows. Two experiments with a replicated 4 × 4 Latin square design were conducted simultaneously with 16 (3 rumen-cannulated) multiparous Holstein cows to measure indicators of ruminal CP degradability (ruminal NH3 concentration, branched-chain volatile fatty acids), metabolizable protein supply (plasma essential AA concentration), N use efficiency (N isotopic discrimination), and dairy performance. In parallel, apparent total-tract digestibility of dry matter, organic matter, neutral detergent fibers, N, total Lys and CML, and partition of N and CML were measured with 4 cows in both experiments. The diets consisted on a DM basis of 20% raw or extruded lupines and 80% basal mixed ration of corn silage, silage and hay from permanent grasslands, pelleted concentrate, and a vitaminized mineral mix. Expected output temperatures of lupine extrusion were 115°C, 135°C, and 150°C, without and with the addition of reducing sugars before extrusion. The extrusion numerically reduced the in vitro ruminal CP degradability of the lupines, and consequently increased the predicted supply of CP to the small intestine. Nitrogen balance and urinary N excretion did not differ among dietary treatments in either experiment. Milk yield and N use efficiency for milk production increased with extrusion of lupines at 150°C without addition of reducing sugars compared with raw lupines. Nitrogen isotopic discrimination between dietary and animal proteins (the difference between δ15N in plasma and δ15N in the diet) were lower with lupines extruded at 150°C without and with addition of reducing sugars. Regardless of sugar addition, milk true protein yield was not affected, but milk urea concentration and fat:protein ratio were lower with lupines extruded at 150°C than with raw lupines. In the CML partition study, we observed that on average 26% of the apparently digested CML was excreted in urine, and a much lower proportion (0.63% on average) of the apparently digested CML was secreted in milk, with no differences among dietary treatments. In conclusion, we showed that the extrusion of white lupines without or with addition of reducing sugars numerically reduced enzymatic CP degradability, with limited effects on N partition, but increased milk yield and N use efficiency at the highest temperature of extrusion without addition of reducing sugars.

Several studies tried to identify digestive determinants of individual variation in feed efficiency between fattening bulls, because of their importance for breeding and management strategies. Most studies focused on single traits or single diet. Little is known about diet-dependent differences in digestive determinants and on their relative importance in distinguishing divergent residual feed intake (RFI) bulls. This research aimed (i) to identify digestive traits that differed between bulls diverging in RFI and fed a maize silage- or a grass silage-based diets; (ii) to highlight the relationships between RFI and digestive traits, and (iii) to explore the hierarchy among digestive traits in discriminating RFI divergent bulls. After an initial RFI test of 84 days on 100 Charolais growing bulls fed two different diets based on grass silage (GS), or maize silage (MS), the 32 most RFI divergent bulls were selected (eight efficient RFI− and eight inefficient RFI+ bulls per diet) and measured thereafter for total tract apparent digestibility and transit rate, enteric gas emissions (CH4 and H2), rumen pH, and feeding behaviour. Rumen particle size and visceral organ and reticulo-omasal orifice (ROO) sizes and rumen and ileum histology were measured at slaughter on the 32 selected extreme RFI bulls. Irrespective of the diet, efficient bulls (RFI−) had lower rumen size, CH4 yield (g/kg DM intake; tendency), lower number of cells in the ileal crypts, tended to have longer time of rumen pH below 5.8 and lower proportion of small size particles in rumen content than non-efficient bulls (RFI+). A long-term test for feed efficiency (197 d on average) was performed on the whole experimental period until slaughter for the 100 animals. The long-term RFI value was negatively related to time spent in activity other than ingestion, rumination, and resting, and positively related (tendency) to the duration of ingestion events, to rumen and abomasum size, irrespective of the diet. Diet-dependent effects were noted: with GS, efficient (RFI−) bulls showed a slower transit rate, whereas with MS, efficient (RFI−) bulls tended to have shorter resting events and a smaller ROO than inefficient bulls (RFI+). The transit rate and the ROO size tended to be positively related, while total tract apparent digestibility of nitrogen was negatively related to long-term RFI value, but only in GS. Rumen size appeared as the most discriminating digestive variable between RFI divergent bulls, but this result should be validated on a larger number of animals and diets.

Protein turnover has been associated to residual feed intake (RFI) in beef cattle. However, this relationship may be confounded by feeding level and affected by the composition of the diet being fed. Our aim was to assess postmortem the protein metabolism signalling pathways in skeletal muscle and liver of 32 Charolais young bulls with extreme RFI phenotypes. Bulls were fed two contrasting diets during the whole fattening period but were subjected to a similar and single nutritional stimulus, induced by their respective concentrate, just prior to slaughter. The key targets were protein degradation (autophagy and ubiquitin) and synthesis signalling pathways through western-blot analysis, as well as hepatic transaminase activity. To ensure a precise assessment of all animals at the same postprandial time, they were provided with a test meal (2.5 kg of either a high-starch and high-protein concentrate or high-fibre and low-protein concentrate) 3 hours prior to slaughter, irrespective of their RFI grouping. Blood and tissues were sampled at the slaughterhouse (3 h and 3 h30 postprandially, respectively). In response to an identical single meal size, efficient RFI animals showed higher (P 0.05). Our findings suggest that, in response to an identical single meal size, efficient RFI animals exhibited lower activation of tissue protein degradation pathways and faster muscle protein synthesis activation compared to their inefficient counterparts. This pattern was observed regardless of the composition of the tested meals.

The production of enteric methane in the gastrointestinal tract of livestock is considered as an energy loss in the equations for estimating energy metabolism in feeding systems. Therefore, the spared energy resulting from specific inhibition of methane emissions should be re-equilibrated with other factors of the equation. And, it is commonly assumed that net energy from feeds increases, thus benefitting production functions, particularly in ruminants due to the important production of methane in the rumen. Notwithstanding, we confirm in this work that inhibition of emissions in ruminants does not transpose into consistent improvements in production. Theoretical calculations of energy flows using experimental data show that the expected improvement in net energy for production is small and difficult to detect under the prevailing, moderate inhibition of methane production (≈25%) obtained using feed additives inhibiting methanogenesis. Importantly, the calculation of energy partitioning using canonical models might not be adequate when methanogenesis is inhibited. There is a lack of information on various parameters that play a role in energy partitioning and that may be affected under provoked abatement of methane. The formula used to calculate heat production based on respiratory exchanges should be validated when methanogenesis is inhibited. Also, a better understanding is needed of the effects of inhibition on fermentation products, fermentation heat, and microbial biomass. Inhibition induces the accumulation of H2, the main substrate used to produce methane, that has no energetic value for the host, and it is not extensively used by the majority of rumen microbes. Currently, the fate of this excess of H2 and its consequences on the microbiota and the host are not well known. All this additional information will provide a better account of energy transactions in ruminants when enteric methanogenesis is inhibited. Based on the available information, it is concluded that the claim that enteric methane inhibition will translate into more feed-efficient animals is not warranted.

ABSTRACT: Estimating the efficiency of N utilization for milk production (MNE) of individual cows at a large scale is difficult, particularly because of the cost of measuring feed intake. Nitrogen isotopic discrimination (Δ15N) between the animal (milk, plasma, or tissues) and its diet has been proposed as a biomarker of the efficiency of N utilization in a range of production systems and ruminant species. The aim of this study was to assess the ability of Δ15N to predict the between-animal variability in MNE in dairy cows using an extensive database. For this, 20 independent experiments conducted as either changeover (n = 14) or continuous (n = 6) trials were available and comprised an initial data set of 1,300 observations. Between-animal variability was defined as the variation observed among cows sharing the same contemporary group (CG; individuals from the same experimental site, sampling period, and dietary treatment). Milk N efficiency was calculated as the ratio between mean milk N (grams of N in milk per day) and mean N intake (grams of N intake per day) obtained from each sampling period, which lasted 9.0 ± 9.9 d (mean ± SD). Samples of milk (n = 604) or plasma (n = 696) and feeds (74 dietary treatments) were analyzed for natural 15N abundance (δ15N), and then the N isotopic discrimination between the animal and the dietary treatment was calculated (Δ15n = δ15Nanimal − δ15Ndiet). Data were analyzed through mixed-effect regression models considering the experiment, sampling period, and dietary treatment as random effects. In addition, repeatability estimates were calculated for each experiment to test the hypothesis of improved predictions when MNE and Δ15N measurements errors were lower. The considerable protein mobilization in early lactation artificially increased both MNE and Δ15N, leading to a positive rather than negative relationship, and this limited the implementation of this biomarker in early lactating cows. When the experimental errors of Δ15N and MNE decreased in a particular experiment (i.e., higher repeatability values), we observed a greater ability of Δ15N to predict MNE at the individual level. The predominant negative and significant correlation between Δ15N and MNE in mid- and late lactation demonstrated that on average Δ15N reflects MNE variations both across dietary treatments and between animals. The root mean squared prediction error as a percentage of average observed value was 6.8%, indicating that the model only allowed differentiation between 2 cows in terms of MNE within a CG if they differed by at least 0.112 g/g of MNE (95% confidence level), and this could represent a limitation in predicting MNE at the individual level. However, the one-way ANOVA performed to test the ability of Δ15N to differentiate within-CG the top 25% from the lowest 25% individuals in terms of MNE was significant, indicating that it is possible to distinguish extreme animals in terms of MNE from their N isotopic signature, which could be useful to group animals for precision feeding.

ABSTRACT: Urine is a highly suitable biological matrix for metabolomics studies. Total collection for 24-h periods is the gold standard as it ensures the presence of all metabolites excreted throughout the day. However, in animal studies, it presents limitations related to animal welfare and also due to alterations of the metabolome originating from the use of acid for preventing microbial growth or microbial contamination. In this study, we investigated whether spot urine collection is a practical alternative to total collection for metabolomic studies in lactating cows. For this purpose, we collected urine samples from 4 lactating Holstein cows fed 4 diets in a 4 × 4 Latin square design. Urine was collected for 24 h using a collecting device (i.e., total collection) or collected once per day 4 h after the morning feeding (i.e., spot urine collection). Dietary treatments differed by the amount of nitrogen content (high vs. low) and by the nature of the energy (starch vs. fiber). Urine metabolome was analyzed by 2 untargeted complementary methods, nuclear magnetic resonance and hydrophilic-interaction liquid chromatography (HILIC) coupled to a time-of-flight mass spectrometer, and by 1 targeted method, HILIC–tandem mass spectrometry. Although sampling technique had an effect on the abundance of metabolites detected, spot urine samples were equally capable of showing differences in urine metabolome than samples from total collection. When considering nitrogen levels in the diet, the robustness and precision for discriminating high- and low-nitrogen diets was equally achieved with both sampling techniques. A total of 22 discriminant metabolites associated with the N level of diets were identified from untargeted HILIC coupled to a time-of-flight mass spectrometer (n = 9) and nuclear magnetic resonance (n = 11), and 2 from targeted HILIC–tandem mass spectrometry. Alternatively, starch or fiber in the diet induced less changes in the metabolome that were not clearly discriminated independently of the sampling technique. We concluded that spot urine collection can successfully reveal differences in the urine metabolome elicited by dietary N levels and be used as a substitute of total urinary 24-h collection for metabolomic studies.

In the current economic and environmental context, the selection of livestock phenotypes combining high feed efficiency (FE) and low greenhouse gas emissions is interesting. This study aimed to quantify methane (CH4) emissions and other gas flows (carbon dioxide (CO2) and dihydrogen (H2) emissions, oxygen (O2) consumption) in growing bulls fed with two contrasting diets in order to (i) evaluate the persistence of individual variability in gas flows through time, and (ii) assess the inter-individual relationship between gas flows and FE across diets. Charolais bulls were fattened for 6 months during two consecutive years in two independent batches (50–51 per year). In each batch, half of the animals received a total mixed ad libitum ration either based on maize silage (62% dietary DM) or high-starch concentrate (MS-S), and half based on grass silage (59% dietary DM) and high-fibre concentrate (GS-F). The absolute gas flows (g/d) were individually measured with 2 GreenFeed systems during 88 days (group 1) and 64 days (group 2). All gas flows were also expressed in g/kg DM intake (gas yield), in g/kg average daily gain (CH4 intensity) and residual of daily emissions for CH4 (R CH4). Different FE metrics (residual feed intake (RFI), residual gain (RG) and feed conversion efficiency (FCE)) were investigated during the same period. The relationships between gas flows and FE metrics were tested by linear regression with the diet as fixed effect. For both diets, we observed a consistent individual variability over the measurement period for absolutes values (g/d) of CH4, CO2, and O2 (repeatability >0.7 for GS-F and >0.6 for MS-S). Gas flows (g/d) were positively correlated with RFI with both diets: animals that ingested food in excess of their theoretical maintenance and growth requirements emitted more CH4, CO2 and consumed more O2. The positive relationship between absolute CH4 emissions and RFI highlighted the interest for low-CH4 emitters and efficient growing bulls when fed with high-energy diets rich in starch or fibre. For both diets, RCH4, CH4 yield and CH4 intensity were not related to RFI whereas a significant negative relationship was reported between CH4 intensity and RG, and FCE. These data suggest that intake is the main driver of the phenotypic relationships between CH4 traits and RFI. Further studies including larger numbers of animals on highly contrasting energy diets are needed to investigate the underlying biological regulatory mechanisms of the methanogenic potential of an animal in relation to production traits.

The objective of this study was to evaluate the effects of oak tannin extract (OTE) added in forage before ensiling on dairy cows fed at 92% of their digestible protein requirements. Six multiparous lactating Holstein cows were used in a crossover design (two treatments × two periods). The control treatment (CON) was based on a diet including 50% of grass silage, whereas the experimental treatment (TAN) included grass silage sprayed with OTE (26 g/kg DM) just before baling. Milk yield (on average 24 kg fat protein corrected milk per day) was not affected, but both milk and rumen fatty acids profiles were impacted by OTE. Nitrogen intake (415 g N per cow per day) and nitrogen use efficiency (NUE; 0.25 on average) were not affected, but a shift from urine (−8% of N intake relatively to control, P = 0.06) to faecal N (+5%; P = 0.004) was observed with the TAN diet (P ≤ 0.05). Nitrogen apparent digestibility was thus reduced for TAN (−3%; P ≤ 0.05). The effect of OTE on ruminal and milk FA profiles suggests an impact on rumen microbiota. Nitrogen isotopic discrimination between animal proteins and diet (Δ15N) was evaluated as a proxy for NUE. While no differences in NUE were observed across diets, a lower Δ15N of plasma proteins was found when comparing TAN v. CON diets. This finding supports the concept that Δ15N would mainly sign the N partitioning at the metabolic level rather than the overall NUE, with the latter also being impacted by digestive processes. Our results agree with a N shift from urine to faeces, and this strategy can thus be adopted to decrease the environmental impact of ruminant protein feeding.

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 (Δ15Nanimal-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 Δ15Nanimal-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 Δ15Nanimal-diet was a useful indicator not only for NUE but also for urinary N excretion, which is a major environmental pollution factor on farm.

Animal nitrogen (N) partitioning is a key parameter for profitability and sustainability of ruminant production systems, which may be predicted from N isotopic discrimination or fractionation (Δ15N). Both animal genetics and feeding level may interact and impact on N partitioning. Therefore, this study aimed to assess the interactive effects of genetic merit (G) and feed allowance (F) on N partitioning and Δ15N in sheep. The sheep were drawn from two levels of G (high G vs. low G; based on New Zealand Sheep Improvement Limited (http://www.sil.co.nz/) dual (wool and meat) growth index) and allocated to two levels of F (1.7 (high F) vs. 1.1 (low F) times Metabolisable Energy requirement for maintenance) treatments. Twenty-four Coopworth rams were divided into four equal groups for a N balance study: high G × high F, high G × low F, low G × high F, and low G × low F. The main factors (G and F) and the interaction term were used for 2-way ANOVA and regression analysis. Higher F led to higher N excretions (urinary N (UN); faecal N (FN); manure N), retained N, N use efficiency (NUE), and urinary purine derivatives excretion (P

Developing the rumen’s capacity to utilise recalcitrant and low-value feed resources is important for ruminant production systems. Early-life nutrition and management practices have been shown to influence development of the rumen in young animals with long-term consequences on their performance. Therefore, there has been increasing interest to understand ruminal development and function in young ruminants to improve feed efficiency, health, welfare, and performance of both young and adult ruminants. However, due to the small size, rapid morphological changes and low initial microbial populations of the rumen, it is difficult to study ruminal function in young ruminants without major invasive approaches or slaughter studies. In this review, we discuss the usefulness of a range of proxies and markers to monitor ruminal function and nitrogen use efficiency (a major part of feed efficiency) in young ruminants. Breath sulphide and methane emissions showed the greatest potential as simple markers of a developing microbiota in young ruminants. However, there is only limited evidence for robust indicators of feed efficiency at this stage. The use of nitrogen isotopic discrimination based on plasma samples appeared to be the most promising proxy for feed efficiency in young ruminants. More research is needed to explore and refine potential proxies and markers to indicate ruminal function and feed efficiency in young ruminants, particularly for neonatal ruminants.

Identifying animals that are superior in terms of feed efficiency may improve the profitability and sustainability of the beef cattle sector. However, measuring feed efficiency is costly and time-consuming. Biomarkers should thus be explored and validated to predict between-animal variation of feed efficiency for both genetic selection and precision feeding. In this work, we aimed to assess and validate two previously identified biomarkers of nitrogen (N) use efficiency in ruminants, plasma urea concentrations and the 15N natural abundance in plasma proteins (plasma δ15N), to predict the between-animal variation in feed efficiency when animals were fed two contrasted diets (high-starch vs high-fibre diets). We used an experimental network design with a total of 588 young bulls tested for feed efficiency through two different traits (feed conversion efficiency [FCE] and residual feed intake [RFI]) during at least 6 months in 12 cohorts (farm × period combination). Animals reared in the same cohort, receiving the same diet and housed in the same pen, were considered as a contemporary group (CG). To analyse between-animal variations and explore relationships between biomarkers and feed efficiency, two statistical approaches, based either on mixed-effect models or regressions from residuals, were conducted to remove the between-CG variability. Between-animal variation of plasma δ15N was significantly correlated with feed efficiency measured through the two criteria traits and regardless of the statistical approach. Conversely, plasma urea was not correlated to FCE and showed only a weak, although significant, correlation with RFI. The response of plasma δ15N to FCE variations was higher when animals were fed a high-starch compared to a high-fibre diet. In addition, we identified two dietary factors, the metabolisable protein to net energy ratio and the rumen protein balance that influenced the relation between plasma δ15N and FCE variations. Concerning the genetic evaluation, and despite the moderate heritability of the two biomarkers (0.28), the size of our experimental setup was insufficient to detect significant genetic correlations between feed efficiency and the biomarkers. However, we validated the potential of plasma δ15N to phenotypically discriminate two animals reared in identical conditions in terms of feed efficiency as long as they differ by at least 0.049 g/g for FCE and 1.67 kg/d for RFI. Altogether, the study showed phenotypic, but non-genetic, relationships between plasma proteins δ15N and feed efficiency that varied according to the efficiency index and the diet utilised.

Residual feed intake (RFI) is an alternative measure of feed efficiency (FE) and is calculated as the difference between actual and expected feed intake. The biological mechanisms underlying animal-to-animal variation in FE are not well understood. The aim of this study was to investigate the digestive ability of beef cows selected for RFI divergence as heifers, using two contrasted diets. Fifteen 4-year-old beef cows were selected from a total of 69 heifers based on their RFI following the feedlot test. The selected heifers were ranked into high-RFI (+ 1.02 ± 0.28, n = 8) and low-RFI (−0.73 ± 0.28, n = 7), and a digestibility trial was performed after their first lactation. Both RFI groups were offered two different diets: 100% hay or a fattening diet which consisted of a DM basis of 67% whole-plant maize silage and 33% high starch concentrates over four experimental periods (two per diet). A diet effect was observed on feed intake and apparent digestibility, whereas no diet × RFI interaction was detected (P > 0.05). Intake and apparent digestibility were higher in cows fed the fattening diet than in those fed the hay diet (P < 0.0001). DM intake (DMI) and organic matter apparent digestibility (OMd) were repeatable and positively correlated between the two subsequent periods of measurements. For the hay and fattening diets, the repeatability between periods was r = 0.71 and r = 0.73 for DMI and r = 0.87 and r = 0.48 for OMd, respectively. Moreover, both intake (r = 0.55) and OMd (r = 0.54) were positively correlated (P < 0.05) between the hay and fattening diets. Significant differences between beef cows selected for divergence in RFI as heifers were observed for digestive traits (P < 0.05), DM and organic matter (OM) apparent digestibility being higher for low-RFI cows. Overall, this study showed that apparent digestibility contributes to between-animal variation in FE in beef cows.

Animal proteins are naturally 15N enriched relative to the diet and the extent of this difference (Δ15Nanimal-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 Δ15Nanimal-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 Δ15Nanimal-diet. Individual values from eight publications reporting both NUE and Δ15Nanimal-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 Δ15Nanimal-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 Δ15Nanimal-diet variation, with the objective of identifying common variables linking NUE and Δ15Nanimal-diet. For individuals reared under similar conditions (within-study) and at the same time (within-period), the variance of NUE and Δ15Nanimal-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 Δ15Nanimal-diet was significantly and negatively correlated to NUE variation across diets (NUE=0.415 −0.055×Δ15Nanimal-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×Δ15Nanimal-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 Δ15Nanimal-diet (without the influence of experimental factors), the negative relationship was still significant, highlighting the ability of Δ15Nanimal-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 Δ15Nanimal-diet and NUE variation, with rumen fermentation and digestion contributing to a lesser extent. This study confirmed that on average Δ15Nanimal-diet can predict NUE variation across diets and across individuals reared under similar conditions.

Animal tissues are naturally 15N enriched relative to their diet and the extent of this difference (Δ15Nanimal-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 Δ15Nanimal-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 (Δ15Nanimal-diet) in these different body pools. The Δ15Nanimal-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 (Δ15Nduodenal digesta-diet mean value=0.42) compared with that observed in plasma proteins (Δ15Nplasma protein-diet mean value=2.41). These data support the idea that most of the isotopic N fractionation observed in ruminant proteins (Δ15Nplasma protein-diet) has a metabolic origin with very little direct impact of the overall digestion process on the existing relationship between Δ15Nplasma 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 (Δ15Nplasma protein-diet) could have a metabolic origin, but more studies are warranted to confirm this point with different diets and approaches.

Animal’s feed efficiency in growing cattle (i.e. the animal ability to reach a market or adult BW with the least amount of feed intake), is a key factor in the beef cattle industry. Feeding systems have made huge progress to understand dietary factors influencing the average animal feed efficiency. However, there exists a considerable amount of animal-to-animal variation around the average feed efficiency observed in beef cattle reared in similar conditions, which is still far from being understood. This review aims to identify biological determinants and molecular pathways involved in the between-animal variation in feed efficiency with particular reference to growing beef cattle phenotyped for residual feed intake (RFI). Moreover, the review attempts to distinguish true potential determinants from those revealed through simple associations or indirectly linked to RFI through their association with feed intake. Most representative and studied biological processes which seem to be connected to feed efficiency were reviewed, such as feeding behaviour, digestion and methane production, rumen microbiome structure and functioning, energy metabolism at the whole body and cellular levels, protein turnover, hormone regulation and body composition. In addition, an overall molecular network analysis was conducted for unravelling networks and their linked functions involved in between-animal variation in feed efficiency. The results from this review suggest that feeding and digestive-related mechanisms could be associated with RFI mainly because they co-vary with feed intake. Although much more research is warranted, especially with high-forage diets, the role of feeding and digestive related mechanisms as true determinants of animal variability in feed efficiency could be minor. Concerning the metabolic-related mechanisms, despite the scarcity of studies using reference methods it seems that feed efficient animals have a significantly lower energy metabolic rate independent of the associated intake reduction. This lower heat production in feed efficient animals may result from a decreased protein turnover and a higher efficiency of ATP production in mitochondria, both mechanisms also identified in the molecular network analysis. In contrast, hormones and body composition could not be conclusively related to animal-to-animal variation in feed efficiency. The analysis of potential biological networks underlying RFI variations highlighted other significant pathways such as lipid metabolism and immunity and stress response. Finally, emerging knowledge suggests that metabolic functions underlying genetic variation in feed efficiency could be associated with other important traits in animal production. This emphasizes the relevance of understanding the biological basis of relevant animal traits to better define future balanced breeding programmes.

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.

Dans le cadre de la révision des unités d’alimentation des ruminants (projet INRA « Systali »), les besoins protéiques ainsi que les réponses aux variations des apports protéiques ont été actualisés. Le calcul des besoins d’entretien intègre les trois principales voies de pertes obligatoires azotées, qui aboutissent aux besoins protéiques non productifs : les protéines endogènes fécales, les pertes azotées urinaires endogènes et les pertes de protéines par les phanères. Ces actualisations ont été effectuées par approche factorielle et à partir de méta-analyses de larges bases de données de la littérature. Les nouvelles valeurs de ces besoins en protéines digestibles dans l’intestin (PDI) « non productifs » ont ensuite été appliquées à de nombreuses expériences conduites sur vaches ou chèvres laitières avec pour objectif d’étudier les variations des réponses aux apports protéiques. Pour l’estimation de la réponse de l’efficacité des PDI en protéine de lait (EffPDI), il a été décidé d’appliquer une valeur commune pour les différentes productions de protéines ainsi que pour l’accrétion et la mobilisation des protéines corporelles, liées au bilan d’énergie. EffPDI est une fonction décroissante non linéaire de la concentration en PDI, identique entre les vaches et les chèvres. Il est aussi possible de prédire la réponse de la production des protéines laitières en fonction du niveau d’apport des PDI disponibles et en utilisant, comme situation pivot, soit la production pour EffPDI = 67%, soit le potentiel de production. Les données recueillies ont également permis de calculer, de façon factorielle, les pertes azotées urinaires dont les deux principales causes sont l’inefficacité des PDI et l’état nutritionnel azoté du rumen évalué par le critère de la balance protéique du rumen (BalProRu).

Identification of plasma biomarkers for feed efficiency in growing beef cattle offers a promising opportunity for developing prediction models to improve precision feeding strategies. However, these models must accurately predict feed efficiency at early stages of fattening. Our study aimed to evaluate the reliability of candidate biomarkers previously identified in late-fattening cattle when analysed during early fattening stages and to develop diet-specific prediction equations for residual feed intake (RFI). From a total of 364 Charolais bulls across seven cohorts, we selected 64 animals with extreme RFI values. The animals were fed either a corn‑ or grass-silage diets. These animals were chosen from four out of the available seven cohorts. Animals from three cohorts (24 high-RFI and 24 low-RFI, having a mean RFI difference of 1.48 kg/d) were used for biomarker confirmation and prediction model training. Animals from a fourth cohort (8 high-RFI and 8 low-RFI, having a mean RFI difference of 0.98 kg/d) were used for model external validation. Blood samples were collected at the beginning of the feed efficiency test (333 ± 20 days), and plasma underwent targeted metabolomic for 630 metabolites, natural abundance of 15 N (δ 15 N), insulin, and IGF-1 analysis. Seven previously identified plasma biomarkers for RFI in late-fattening beef cattle still kept their capability for discriminating low and high RFI animals when analysed during early fattening stages (P < 0.05). Among these confirmed biomarkers, five were common for both grass- and corn-fed animals (creatinine, β-alanine, triglyceride TG18:0&#95;34:2, symmetric dimethyl-arginine and phosphatidylcholine PC aa C30:2) while two were diet-specific (IGF-1 for grass silage-based diet, and isoleucine for corn silage-based diet. No new plasma biomarkers of RFI were identified at early-fattening stages (false discovery rate  > 0.05). Prediction models were developed based on seven confirmed RFI biomarkers analysed during early-fattening. Two logistic regression models incorporating creatinine and either IGF-1 (for grass silage-based diet) or PC aa C30:2 (for corn silage-based diet) effectively distinguished between high- and low-RFI animals with high sensitivity and specificity (area under the curve > 0.80). The biomarkers used in the models showed moderate to high repeatability between early and late fattening stages (0.45 < r < 0.65). The models were successfully externally validated, with more than 85% of animals from the fourth cohort correctly classified. Once validated in larger cohorts and utilising cost-effective and rapid analytical methods, these models could support precision feeding and breeding programmes, aiming to reduce the cost of raising beef cattle., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

The selection of more efficient animals for breeding is of both economic and environmental interest to the industry. The aim of this study was to evaluate the influence of the animals' residual feed intake (RFI) ranking in interaction with the type of diet on the meat quality of Charolais beef cattle. Indeed, several biological mechanisms are associated with RFI, especially when animals are fed high starch-diets. It is therefore possible that quality parameters may show greater changes due to RFI in the context of high starch diets compared to high forage diets. An 84-day feed efficiency trial followed immediately by a second 112-day feed efficiency trial was conducted with a total of 100 animals fed either maize- or grass-diets for 196-days. At the end of the 84-day period, the 32 most divergent RFI animals (16 extreme RFI animals per diet, 8 RFI+ and 8 RFI-) were identified. They were slaughtered after 112-days of finishing. The Longissimus thoracis was characterised in terms of nutritional and sensory quality. RFI had no effect on lab colour, muscle shear force, total fat, fatty acid ratios and most of the total fatty acid content (especially n-3) irrespective of the diet. However, more efficient animals (RFI-) showed higher CLA contents compared to less efficient animals (RFI+) regardless of the diet and also a lower n6/n3 ratio only in animals fed the maize diets. Diet also had a significant effect on lipid and FA content as well as on FA composition., Competing Interests: Declaration of competing interest None., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

This study investigated whether plasma biomarkers of residual feed intake (RFI), identified under ad libitum feeding conditions in beef cattle, remained consistent during feed restriction. Sixty Charolais crossbred young bulls were divided into two groups for a crossover study. Group A was initially fed ad libitum (first test) and then restricted (second test) on the same diet, while Group B experienced the opposite sequence. Blood samples were collected from the 12 most divergent RFI animals in each group at the end of the first test and again after the second test. 12 plasma variables consistently increased, while three consistently decreased during feed restriction (FDR < 0.05). Only two metabolites, α-aminoadipic acid for Group A and 5-aminovaleric acid for Group B, were associated with RFI independent of feed intake level (FDR < 0.05), demonstrating moderate-to-high repeatability across feeding levels (intraclass correlation coefficient ≥ 0.59). Notably, both metabolites belong to the same metabolic pathway: lysine degradation. These metabolites consistently correlated with RFI, irrespective of fluctuations in feed intake, indicating a connection to individual metabolic processes influencing feed efficiency. These findings suggest that a portion of RFI phenotypic variance is inherent to an individual's metabolic efficiency beyond variations in feed intake., (© 2024. The Author(s).)

Several studies tried to identify digestive determinants of individual variation in feed efficiency between fattening bulls, because of their importance for breeding and management strategies. Most studies focused on single traits or single diet. Little is known about diet-dependent differences in digestive determinants and on their relative importance in distinguishing divergent residual feed intake (RFI) bulls. This research aimed (i) to identify digestive traits that differed between bulls diverging in RFI and fed a maize silage- or a grass silage-based diets; (ii) to highlight the relationships between RFI and digestive traits, and (iii) to explore the hierarchy among digestive traits in discriminating RFI divergent bulls. After an initial RFI test of 84 days on 100 Charolais growing bulls fed two different diets based on grass silage (GS), or maize silage (MS), the 32 most RFI divergent bulls were selected (eight efficient RFI- and eight inefficient RFI+ bulls per diet) and measured thereafter for total tract apparent digestibility and transit rate, enteric gas emissions (CH 4 and H 2 ), rumen pH, and feeding behaviour. Rumen particle size and visceral organ and reticulo-omasal orifice (ROO) sizes and rumen and ileum histology were measured at slaughter on the 32 selected extreme RFI bulls. Irrespective of the diet, efficient bulls (RFI-) had lower rumen size, CH 4 yield (g/kg DM intake; tendency), lower number of cells in the ileal crypts, tended to have longer time of rumen pH below 5.8 and lower proportion of small size particles in rumen content than non-efficient bulls (RFI+). A long-term test for feed efficiency (197 d on average) was performed on the whole experimental period until slaughter for the 100 animals. The long-term RFI value was negatively related to time spent in activity other than ingestion, rumination, and resting, and positively related (tendency) to the duration of ingestion events, to rumen and abomasum size, irrespective of the diet. Diet-dependent effects were noted: with GS, efficient (RFI-) bulls showed a slower transit rate, whereas with MS, efficient (RFI-) bulls tended to have shorter resting events and a smaller ROO than inefficient bulls (RFI+). The transit rate and the ROO size tended to be positively related, while total tract apparent digestibility of nitrogen was negatively related to long-term RFI value, but only in GS. Rumen size appeared as the most discriminating digestive variable between RFI divergent bulls, but this result should be validated on a larger number of animals and diets., (Copyright © 2023. Published by Elsevier B.V.)

The extrusion of leguminous seeds induces the formation of Maillard reaction compounds (MRC) as a product of protein advanced glycation and oxidation, which lowers protein degradability in the rumen. However, the quantitative relationship between the parameters of pretreatment (i.e., addition of reducing sugars) and extrusion, and the formation of MRC has not been established yet. Moreover, the fate of the main stable MRC, Nε-carboxymethyl-lysine (CML), in the excretory routes has never been investigated in ruminants. We aimed to test the effects of the temperature of extrusion of white lupines with or without addition of reducing sugars on the formation of MRC, crude protein (CP) degradability in the rumen, N use efficiency for milk production (milk N/N intake), and performance of dairy cows. Two experiments with a replicated 4 × 4 Latin square design were conducted simultaneously with 16 (3 rumen-cannulated) multiparous Holstein cows to measure indicators of ruminal CP degradability (ruminal NH 3 concentration, branched-chain volatile fatty acids), metabolizable protein supply (plasma essential AA concentration), N use efficiency (N isotopic discrimination), and dairy performance. In parallel, apparent total-tract digestibility of dry matter, organic matter, neutral detergent fibers, N, total Lys and CML, and partition of N and CML were measured with 4 cows in both experiments. The diets consisted on a DM basis of 20% raw or extruded lupines and 80% basal mixed ration of corn silage, silage and hay from permanent grasslands, pelleted concentrate, and a vitaminized mineral mix. Expected output temperatures of lupine extrusion were 115°C, 135°C, and 150°C, without and with the addition of reducing sugars before extrusion. The extrusion numerically reduced the in vitro ruminal CP degradability of the lupines, and consequently increased the predicted supply of CP to the small intestine. Nitrogen balance and urinary N excretion did not differ among dietary treatments in either experiment. Milk yield and N use efficiency for milk production increased with extrusion of lupines at 150°C without addition of reducing sugars compared with raw lupines. Nitrogen isotopic discrimination between dietary and animal proteins (the difference between δ 15 N in plasma and δ 15 N in the diet) were lower with lupines extruded at 150°C without and with addition of reducing sugars. Regardless of sugar addition, milk true protein yield was not affected, but milk urea concentration and fat:protein ratio were lower with lupines extruded at 150°C than with raw lupines. In the CML partition study, we observed that on average 26% of the apparently digested CML was excreted in urine, and a much lower proportion (0.63% on average) of the apparently digested CML was secreted in milk, with no differences among dietary treatments. In conclusion, we showed that the extrusion of white lupines without or with addition of reducing sugars numerically reduced enzymatic CP degradability, with limited effects on N partition, but increased milk yield and N use efficiency at the highest temperature of extrusion without addition of reducing sugars., (© 2023, The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Protein turnover has been associated to residual feed intake (RFI) in beef cattle. However, this relationship may be confounded by feeding level and affected by the composition of the diet being fed. Our aim was to assess postmortem the protein metabolism signalling pathways in skeletal muscle and liver of 32 Charolais young bulls with extreme RFI phenotypes. Bulls were fed two contrasting diets during the whole fattening period but were subjected to a similar and single nutritional stimulus, induced by their respective concentrate, just prior to slaughter. The key targets were protein degradation (autophagy and ubiquitin) and synthesis signalling pathways through western-blot analysis, as well as hepatic transaminase activity. To ensure a precise assessment of all animals at the same postprandial time, they were provided with a test meal (2.5 kg of either a high-starch and high-protein concentrate or high-fibre and low-protein concentrate) 3 hours prior to slaughter, irrespective of their RFI grouping. Blood and tissues were sampled at the slaughterhouse (3 h and 3 h30 postprandially, respectively). In response to an identical single meal size, efficient RFI animals showed higher (P < 0.05) postprandial plasma β-hydroxybutyrate concentrations and insulinemia (only with the high-starch concentrate) than non-efficient animals. Moreover, efficient RFI bulls had lower muscle (P = 0.04) and liver (P = 0.08) ubiquitin protein abundance (degradation pathway) and tended to have lower alanine transaminase activity in the liver (P = 0.06) compared to non-efficient bulls, regardless of diet. A positive correlation between protein degradation potential and amino acid catabolism was identified in this study (r = 0.52, P = 0.004), which was interpreted as being biologically linked to the RFI phenotype. Efficient RFI bulls also had a faster potential for protein synthesis in the muscle, as indicated by their greater ratio of phosphorylated to total form of ribosomal protein S6 kinase (P = 0.05), regardless of diet. Results on protein synthesis pathway in muscle and plasma metabolite concentrations suggested that efficient RFI cattle may have a faster nutrient absorption and insulin responsiveness after feeding than inefficient cattle. We did not find significant differences in hepatic protein synthesis pathways between the two RFI groups (P > 0.05). Our findings suggest that, in response to an identical single meal size, efficient RFI animals exhibited lower activation of tissue protein degradation pathways and faster muscle protein synthesis activation compared to their inefficient counterparts. This pattern was observed regardless of the composition of the tested meals., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

Nitrogen use efficiency is an important index in ruminants and can be indirectly evaluated through the N isotopic discrimination between the animal and its diet (Δ15Nanimal-diet). The concentration and source of N may determine both the extent of the N isotopic discrimination in bacteria and N use efficiency. We hypothesised that the uptake and release of ammonia by rumen bacteria will affect the natural 15N enrichment of the bacterial biomass over their substrates (Δ15Nbacteria-substrate) and thereby further impacting Δ15Nanimal-diet. To test this hypothesis, two independent in vitro experiments were conducted using two contrasting N sources (organic vs inorganic) at different levels either in pure rumen bacteria culture incubations (Experiment #1) or in mixed rumen cultures (Experiment #2). In Experiment #1, tryptone casein or ammonium chloride were tested at low (1 mM N) and high (11.5 mM N) concentrations on three rumen bacterial strains (Fibrobacter succinogenes, Eubacterium limosum and Xylanibacter ruminicola) incubated in triplicate in anaerobic batch monocultures during 48h. In Experiment #2 mixed rumen cultures were incubated during 120 h with peptone or ammonium chloride at five different levels of N (1.5, 3, 4.5, 6 and 12-mM). In experiment #1, Δ15Nbacteria-substrate was lowest when the ammonia-consumer bacterium Fibrobacter succinogenes was grown on ammonium chloride, and highest when the proteolytic bacterial strain Xylanibacter ruminicola was grown on tryptone. In experiment #2, Δ15Nbacteria-substrate was lower with inorganic (ammonium chloride) vs organic (peptone) N source. A strong negative correlation between Δ15Nbacteria-substrate and Rikenellaceae&#95;RC9&#95;gut&#95;group, a potential fibrolytic rumen bacterium, was detected. Together, our results showed that Δ15Nbacteria-substrate may change according to the balance between synthesis of microbial protein from ammonia versus non-ammonia N sources and confirm the key role of rumen bacteria as modulators of Δ15Nanimal-diet., Competing Interests: The authors declare no conflict of interest., (Copyright: © 2023 Cantalapiedra-Hijar et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The production of enteric methane in the gastrointestinal tract of livestock is considered as an energy loss in the equations for estimating energy metabolism in feeding systems. Therefore, the spared energy resulting from specific inhibition of methane emissions should be re-equilibrated with other factors of the equation. And, it is commonly assumed that net energy from feeds increases, thus benefitting production functions, particularly in ruminants due to the important production of methane in the rumen. Notwithstanding, we confirm in this work that inhibition of emissions in ruminants does not transpose into consistent improvements in production. Theoretical calculations of energy flows using experimental data show that the expected improvement in net energy for production is small and difficult to detect under the prevailing, moderate inhibition of methane production (≈25%) obtained using feed additives inhibiting methanogenesis. Importantly, the calculation of energy partitioning using canonical models might not be adequate when methanogenesis is inhibited. There is a lack of information on various parameters that play a role in energy partitioning and that may be affected under provoked abatement of methane. The formula used to calculate heat production based on respiratory exchanges should be validated when methanogenesis is inhibited. Also, a better understanding is needed of the effects of inhibition on fermentation products, fermentation heat, and microbial biomass. Inhibition induces the accumulation of H 2 , the main substrate used to produce methane, that has no energetic value for the host, and it is not extensively used by the majority of rumen microbes. Currently, the fate of this excess of H 2 and its consequences on the microbiota and the host are not well known. All this additional information will provide a better account of energy transactions in ruminants when enteric methanogenesis is inhibited. Based on the available information, it is concluded that the claim that enteric methane inhibition will translate into more feed-efficient animals is not warranted., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

This study explored plasma biomarkers and metabolic pathways underlying feed efficiency measured as residual feed intake (RFI) in Charolais heifers. A total of 48 RFI extreme individuals (High-RFI, n = 24; Low-RFI, n = 24) were selected from a population of 142 heifers for classical plasma metabolite and hormone quantification and plasma metabolomic profiling through untargeted LC-MS. Most efficient heifers (Low-RFI) had greater (P = 0.03) plasma concentrations of IGF-1 and tended to have (P = 0.06) a lower back fat depth compared to least efficient heifers. However, no changes were noted (P ≥ 0.10) for plasma concentrations of glucose, insulin, non-esterified fatty acids, β-hydroxybutyrate and urea. The plasma metabolomic dataset comprised 3,457 ions with none significantly differing between RFI classes after false discovery rate correction (FDR > 0.10). Among the 101 ions having a raw P < 0.05 for the RFI effect, 13 were putatively annotated by using internal databases and 6 compounds were further confirmed with standards. Metabolic pathway analysis from these 6 confirmed compounds revealed that the branched chain amino acid metabolism was significantly (FDR < 0.05) impacted by the RFI classes. Our results confirmed for the first time in beef heifers previous findings obtained in male beef cattle and pointing to changes in branched-chain amino acids metabolism along with that of body composition as biological mechanisms related to RFI. Further studies are warranted to ascertain whether there is a cause-and-effect relationship between these mechanisms and RFI., Competing Interests: None, (Copyright: © 2022 Jorge-Smeding et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

In the current economic and environmental context, the selection of livestock phenotypes combining high feed efficiency (FE) and low greenhouse gas emissions is interesting. This study aimed to quantify methane (CH 4 ) emissions and other gas flows (carbon dioxide (CO 2 ) and dihydrogen (H 2 ) emissions, oxygen (O 2 ) consumption) in growing bulls fed with two contrasting diets in order to (i) evaluate the persistence of individual variability in gas flows through time, and (ii) assess the inter-individual relationship between gas flows and FE across diets. Charolais bulls were fattened for 6 months during two consecutive years in two independent batches (50-51 per year). In each batch, half of the animals received a total mixed ad libitum ration either based on maize silage (62% dietary DM) or high-starch concentrate (MS-S), and half based on grass silage (59% dietary DM) and high-fibre concentrate (GS-F). The absolute gas flows (g/d) were individually measured with 2 GreenFeed systems during 88 days (group 1) and 64 days (group 2). All gas flows were also expressed in g/kg DM intake (gas yield), in g/kg average daily gain (CH 4 intensity) and residual of daily emissions for CH 4 (R CH 4 ). Different FE metrics (residual feed intake (RFI), residual gain (RG) and feed conversion efficiency (FCE)) were investigated during the same period. The relationships between gas flows and FE metrics were tested by linear regression with the diet as fixed effect. For both diets, we observed a consistent individual variability over the measurement period for absolutes values (g/d) of CH 4, CO 2 , and O 2 (repeatability >0.7 for GS-F and >0.6 for MS-S). Gas flows (g/d) were positively correlated with RFI with both diets: animals that ingested food in excess of their theoretical maintenance and growth requirements emitted more CH 4 , CO 2 and consumed more O 2 . The positive relationship between absolute CH 4 emissions and RFI highlighted the interest for low-CH 4 emitters and efficient growing bulls when fed with high-energy diets rich in starch or fibre. For both diets, RCH 4 , CH 4 yield and CH 4 intensity were not related to RFI whereas a significant negative relationship was reported between CH 4 intensity and RG, and FCE. These data suggest that intake is the main driver of the phenotypic relationships between CH 4 traits and RFI. Further studies including larger numbers of animals on highly contrasting energy diets are needed to investigate the underlying biological regulatory mechanisms of the methanogenic potential of an animal in relation to production traits., (Copyright © 2022 The Author(s). Published by Elsevier B.V. All rights reserved.)

Protein metabolism and body composition have been identified as major determinants of residual feed intake (RFI) in beef cattle fed high-starch fattening diets. This study aimed to evaluate if these two identified RFI determinants in beef cattle are the same across two contrasting silage-based diets. During two consecutive years, an 84-day feed efficiency test (Test A) immediately followed by a second 112-day feed efficiency test (Test B) was carried out using a total of 100 animals offered either one of two diets (either corn silage- or grass silage-based) over 196 days. At the end of Test A, the 32 animals most divergent for RFI (16 extreme RFI animals per diet, eight low RFI and eight high RFI) were identified and evaluated during Test B for their i) N use efficiency (NUE; N retention/N intake) calculated either from a 10-d nitrogen balance trial or from estimations based on body composition changes occurring during the whole experiment (Test A and Test B; 196 days), ii) carcass and whole-body protein turnover rates analysed through the 3-methyl-histidine urinary excretion and the N isotopic turnover rates of urine, respectively, and iii) body composition measured at the slaughterhouse at the end of Test B. Oxygen consumption was measured during Test B for the 100 animals by two GreenFeed systems. Irrespective of the diet, efficient RFI animals tended (P = 0.08) to improve their NUE when N retention was estimated for 196 days or when considering their lower urinary urea-N to total N ratio (P = 0.03). In contrast, NUE calculated during the 10-d N balance showed no differences (P = 0.65) across RFI groups suggesting that this method may not be suitable to capture small NUE differences. Efficient RFI individuals presented higher dressing percentage and muscle deposition in the carcass (P = 0.003) but lighter rumen (P = 0.001), and a trend for lower oxygen consumption (P = 0.08) than inefficient RFI animals irrespective of the diet. Lower protein degradation rates of skeletal muscle and lower protein synthesis rates of plasma proteins were found in efficient RFI cattle but only with the corn silage-based diet (RFI × Diet; P = 0.02). The higher insulinaemia associated with the corn silage-based diet (P = 0.001) seemed to be a key metabolic feature explaining the positive association between protein turnover and RFI only in this diet. Feed N was more efficiently used for growth by efficient RFI animals regardless of the diet but lower protein turnover rates in efficient RFI animals were only observed with corn silage-based diets., (Copyright © 2022 The Author(s). Published by Elsevier B.V. All rights reserved.)

Estimating the efficiency of N utilization for milk production (MNE) of individual cows at a large scale is difficult, particularly because of the cost of measuring feed intake. Nitrogen isotopic discrimination (Δ 15 N) between the animal (milk, plasma, or tissues) and its diet has been proposed as a biomarker of the efficiency of N utilization in a range of production systems and ruminant species. The aim of this study was to assess the ability of Δ 15 N to predict the between-animal variability in MNE in dairy cows using an extensive database. For this, 20 independent experiments conducted as either changeover (n = 14) or continuous (n = 6) trials were available and comprised an initial data set of 1,300 observations. Between-animal variability was defined as the variation observed among cows sharing the same contemporary group (CG; individuals from the same experimental site, sampling period, and dietary treatment). Milk N efficiency was calculated as the ratio between mean milk N (grams of N in milk per day) and mean N intake (grams of N intake per day) obtained from each sampling period, which lasted 9.0 ± 9.9 d (mean ± SD). Samples of milk (n = 604) or plasma (n = 696) and feeds (74 dietary treatments) were analyzed for natural 15 N abundance (δ 15 N), and then the N isotopic discrimination between the animal and the dietary treatment was calculated (Δ 15 n = δ 15 N animal - δ 15 N diet ). Data were analyzed through mixed-effect regression models considering the experiment, sampling period, and dietary treatment as random effects. In addition, repeatability estimates were calculated for each experiment to test the hypothesis of improved predictions when MNE and Δ 15 N measurements errors were lower. The considerable protein mobilization in early lactation artificially increased both MNE and Δ 15 N, leading to a positive rather than negative relationship, and this limited the implementation of this biomarker in early lactating cows. When the experimental errors of Δ 15 N and MNE decreased in a particular experiment (i.e., higher repeatability values), we observed a greater ability of Δ 15 N to predict MNE at the individual level. The predominant negative and significant correlation between Δ 15 N and MNE in mid- and late lactation demonstrated that on average Δ 15 N reflects MNE variations both across dietary treatments and between animals. The root mean squared prediction error as a percentage of average observed value was 6.8%, indicating that the model only allowed differentiation between 2 cows in terms of MNE within a CG if they differed by at least 0.112 g/g of MNE (95% confidence level), and this could represent a limitation in predicting MNE at the individual level. However, the one-way ANOVA performed to test the ability of Δ 15 N to differentiate within-CG the top 25% from the lowest 25% individuals in terms of MNE was significant, indicating that it is possible to distinguish extreme animals in terms of MNE from their N isotopic signature, which could be useful to group animals for precision feeding., (© 2022, The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Urine is a highly suitable biological matrix for metabolomics studies. Total collection for 24-h periods is the gold standard as it ensures the presence of all metabolites excreted throughout the day. However, in animal studies, it presents limitations related to animal welfare and also due to alterations of the metabolome originating from the use of acid for preventing microbial growth or microbial contamination. In this study, we investigated whether spot urine collection is a practical alternative to total collection for metabolomic studies in lactating cows. For this purpose, we collected urine samples from 4 lactating Holstein cows fed 4 diets in a 4 × 4 Latin square design. Urine was collected for 24 h using a collecting device (i.e., total collection) or collected once per day 4 h after the morning feeding (i.e., spot urine collection). Dietary treatments differed by the amount of nitrogen content (high vs. low) and by the nature of the energy (starch vs. fiber). Urine metabolome was analyzed by 2 untargeted complementary methods, nuclear magnetic resonance and hydrophilic-interaction liquid chromatography (HILIC) coupled to a time-of-flight mass spectrometer, and by 1 targeted method, HILIC-tandem mass spectrometry. Although sampling technique had an effect on the abundance of metabolites detected, spot urine samples were equally capable of showing differences in urine metabolome than samples from total collection. When considering nitrogen levels in the diet, the robustness and precision for discriminating high- and low-nitrogen diets was equally achieved with both sampling techniques. A total of 22 discriminant metabolites associated with the N level of diets were identified from untargeted HILIC coupled to a time-of-flight mass spectrometer (n = 9) and nuclear magnetic resonance (n = 11), and 2 from targeted HILIC-tandem mass spectrometry. Alternatively, starch or fiber in the diet induced less changes in the metabolome that were not clearly discriminated independently of the sampling technique. We concluded that spot urine collection can successfully reveal differences in the urine metabolome elicited by dietary N levels and be used as a substitute of total urinary 24-h collection for metabolomic studies., (© 2022, The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Residual feed intake (RFI) is one of the preferred traits for feed efficiency animal breeding. However, RFI measurement is expensive and time-consuming and animal ranking may depend on the nature of the diets. We aimed to explore RFI plasma biomarkers and to unravel the underlying metabolic pathways in yearling bulls fed either a corn-silage diet rich in starch (corn diet) or a grass-silage diet rich in fiber (grass diet). Forty-eight extreme RFI animals (Low-RFI, n = 24, versus High-RFI, n = 24, balanced per diet) were selected from a population of 364 Charolais bulls and their plasma was subjected to a targeted LC-MS metabolomic approach together with classical metabolite and hormonal plasma analyses. Greater lean body mass and nitrogen use efficiency, and lower protein turnover were identified as common mechanisms underlying RFI irrespective of the diet. On the other hand, greater adiposity and plasma concentrations of branched-chain amino acids (BCAA) together with lower insulin sensitivity in High-RFI animals were only observed with corn diet. Conversely, greater plasma concentrations of BCAA and total triglycerides, but similar insulin concentrations were noted in efficient RFI cattle with grass diet. Our data suggest that there are diet-specific mechanisms explaining RFI differences in fattening Charolais yearling bulls., (© 2021. The Author(s).)

Animal nitrogen (N) partitioning is a key parameter for profitability and sustainability of ruminant production systems, which may be predicted from N isotopic discrimination or fractionation (Δ 15 N). Both animal genetics and feeding level may interact and impact on N partitioning. Therefore, this study aimed to assess the interactive effects of genetic merit (G) and feed allowance (F) on N partitioning and Δ 15 N in sheep. The sheep were drawn from two levels of G (high G vs. low G; based on New Zealand Sheep Improvement Limited (http://www.sil.co.nz/) dual (wool and meat) growth index) and allocated to two levels of F (1.7 (high F) vs. 1.1 (low F) times Metabolisable Energy requirement for maintenance) treatments. Twenty-four Coopworth rams were divided into four equal groups for a N balance study: high G × high F, high G × low F, low G × high F, and low G × low F. The main factors (G and F) and the interaction term were used for 2-way ANOVA and regression analysis. Higher F led to higher N excretions (urinary N (UN); faecal N (FN); manure N), retained N, N use efficiency (NUE), and urinary purine derivatives excretion (P < 0.05). On the other hand, higher UN/N intake, and plasma Δ 15 N were observed with the lower F (P < 0.05). Higher G led to increased UN, FN, manure N, apparent N digestibility, and urinary purine derivatives excretion (P < 0.05). Higher F only increased UN in high G sheep, with no effect on low G sheep (P < 0.05). Regression analysis results demonstrated potential to use plasma Δ 15 N to reflect the effects of G and F on NUE and UN/N intake. Further research is urged to study interactive effects of genetic and feeding level on sheep N partitioning., (Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.)

In pasture-based systems, there are nutritional and climatic challenges exacerbated across lactation; thus, dairy cows require an enhanced adaptive capacity compared with cows in confined systems. We aimed to evaluate the effect of lactation stage (21 vs. 180 days in milk, DIM) and Holstein genetic strain (North American Holstein, NAH, n = 8; New Zealand Holstein, NZH, n = 8) on metabolic adaptations of grazing dairy cows through plasma metabolomic profiling and its association with classical metabolites. Although 67 metabolites were affected (FDR < 0.05) by DIM, no metabolite was observed to differ between genetic strains while only alanine was affected (FDR = 0.02) by the interaction between genetic strain and DIM. However, complementary tools for time-series analysis (ASCA analysis, MEBA ranking) indicated that alanine and the branched-chain amino acids (BCAA) differed between genetic strains in a lactation-stage dependent manner. Indeed, NZH cows had lower (P-Tukey < 0.05) plasma concentrations of leucine, isoleucine and valine than NAH cows at 21 DIM, probably signaling for greater insulin sensitivity. Metabolic pathway analysis also revealed that, independently of genetic strains, AA metabolism might be structurally involved in homeorhetic changes as 40% (19/46) of metabolic pathways differentially expressed (FDR < 0.05) between 21 and 180 DIM belonged to AA metabolism., (© 2021. The Author(s).)

Developing the rumen's capacity to utilise recalcitrant and low-value feed resources is important for ruminant production systems. Early-life nutrition and management practices have been shown to influence development of the rumen in young animals with long-term consequences on their performance. Therefore, there has been increasing interest to understand ruminal development and function in young ruminants to improve feed efficiency, health, welfare, and performance of both young and adult ruminants. However, due to the small size, rapid morphological changes and low initial microbial populations of the rumen, it is difficult to study ruminal function in young ruminants without major invasive approaches or slaughter studies. In this review, we discuss the usefulness of a range of proxies and markers to monitor ruminal function and nitrogen use efficiency (a major part of feed efficiency) in young ruminants. Breath sulphide and methane emissions showed the greatest potential as simple markers of a developing microbiota in young ruminants. However, there is only limited evidence for robust indicators of feed efficiency at this stage. The use of nitrogen isotopic discrimination based on plasma samples appeared to be the most promising proxy for feed efficiency in young ruminants. More research is needed to explore and refine potential proxies and markers to indicate ruminal function and feed efficiency in young ruminants, particularly for neonatal ruminants., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

Identifying animals that are superior in terms of feed efficiency may improve the profitability and sustainability of the beef cattle sector. However, measuring feed efficiency is costly and time-consuming. Biomarkers should thus be explored and validated to predict between-animal variation of feed efficiency for both genetic selection and precision feeding. In this work, we aimed to assess and validate two previously identified biomarkers of nitrogen (N) use efficiency in ruminants, plasma urea concentrations and the 15 N natural abundance in plasma proteins (plasma δ 15 N), to predict the between-animal variation in feed efficiency when animals were fed two contrasted diets (high-starch vs high-fibre diets). We used an experimental network design with a total of 588 young bulls tested for feed efficiency through two different traits (feed conversion efficiency [FCE] and residual feed intake [RFI]) during at least 6 months in 12 cohorts (farm × period combination). Animals reared in the same cohort, receiving the same diet and housed in the same pen, were considered as a contemporary group (CG). To analyse between-animal variations and explore relationships between biomarkers and feed efficiency, two statistical approaches, based either on mixed-effect models or regressions from residuals, were conducted to remove the between-CG variability. Between-animal variation of plasma δ 15 N was significantly correlated with feed efficiency measured through the two criteria traits and regardless of the statistical approach. Conversely, plasma urea was not correlated to FCE and showed only a weak, although significant, correlation with RFI. The response of plasma δ 15 N to FCE variations was higher when animals were fed a high-starch compared to a high-fibre diet. In addition, we identified two dietary factors, the metabolisable protein to net energy ratio and the rumen protein balance that influenced the relation between plasma δ 15 N and FCE variations. Concerning the genetic evaluation, and despite the moderate heritability of the two biomarkers (0.28), the size of our experimental setup was insufficient to detect significant genetic correlations between feed efficiency and the biomarkers. However, we validated the potential of plasma δ 15 N to phenotypically discriminate two animals reared in identical conditions in terms of feed efficiency as long as they differ by at least 0.049 g/g for FCE and 1.67 kg/d for RFI. Altogether, the study showed phenotypic, but non-genetic, relationships between plasma proteins δ 15 N and feed efficiency that varied according to the efficiency index and the diet utilised., (Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.)

Background: French beef producers suffer from the decrease in profitability of their farms mainly because of the continuous increase in feed costs. Selection for feed efficiency in beef cattle represents a relevant solution to face this problem. However, feed efficiency is a complex trait that can be assessed by three major criteria: residual feed intake (RFI), residual gain (RG) and feed efficiency ratio (FE), which involve different genetic determinisms. An analysis that combines phenotype and whole-genome sequence data provides a unique framework for genomic studies. The aim of our study was to identify the gene networks and the biological processes that are responsible for the genetic determinism that is shared between these three feed efficiency criteria., Results: A population of 1477 French Charolais young bulls was phenotyped for feed intake (FI), average daily gain (ADG) and final weight (FW) to estimate RFI, RG and FE. A subset of 789 young bulls was genotyped on the BovineSNP50 single nucleotide polymorphism (SNP) array and imputed at the sequence level using RUN6 of the 1000 Bull Genomes Project. We conducted a genome-wide association study (GWAS) to estimate the individual effect of 8.5 million SNPs and applied an association weight matrix (AWM) approach to analyse the results, one for each feed efficiency criterion. The results highlighted co-association networks including 626 genes for RFI, 426 for RG and 564 for FE. Enrichment assessment revealed the biological processes that show the strongest association with RFI, RG and FE, i.e. digestive tract (salivary, gastric and mucin secretion) and metabolic processes (cellular and cardiovascular). Energetic functions were more associated with RFI and FE and cardio-vascular and cellular processes with RG. Several hormones such as apelin, glucagon, insulin, aldosterone, the gonadotrophin releasing hormone and the thyroid hormone were also identified, and these should be tested in future studies as candidate biomarkers for feed efficiency., Conclusions: The combination of network and pathway analyses at the sequence level led to the identification of both common and specific mechanisms that are involved in RFI, RG and FE, and to a better understanding of the genetic determinism underlying these three criteria. The effects of the genes involved in each of the identified processes need to be tested in genomic evaluations to confirm the potential gain in reliability of using functional variants to select animals for feed efficiency.