Liver and Muscle Transcriptomes Differ in Mid-Lactation Cows Divergent in Feed Efficiency in the Presence or Absence of Supplemental Rumen-Protected Choline.

Improving dairy cow feed efficiency is critical to the sustainability and profitability of dairy production, yet the underlying mechanisms that contribute to individual cow variation in feed efficiency are not fully understood. The objectives of this study were to (1) identify genes and associated pathways that are altered in cows with high- or low-residual feed intake (RFI) using RNA sequencing, and (2) determine if rumen-protected choline supplementation during mid-lactation would influence performance or feed efficiency. Mid-lactation (134 ± 20 days in milk) multiparous Holstein cows were randomly assigned to either supplementation of 0 g/d supplementation (CTL; n = 32) or 30 g/d of a rumen-protected choline product (RPC; 13.2 g choline ion; n = 32; Balchem Corp., New Hampton, NY, USA). Residual feed intake was determined as dry matter intake regressed on milk energy output, days in milk, body weight change, metabolic body weight, and dietary treatment. The 12 cows with the highest RFI (low feed efficient; LE) and 12 cows with the lowest RFI (high feed efficient; HE), balanced by dietary treatment, were selected for blood, liver, and muscle analysis. No differences in production or feed efficiency were detected with RPC supplementation, although albumin was greater and arachidonic acid tended to be greater in RPC cows. Concentrations of ß-hydroxybutyrate were greater in HE cows. Between HE and LE, 268 and 315 differentially expressed genes in liver and muscle tissue, respectively, were identified through RNA sequencing. Pathway analysis indicated differences in cell cycling, oxidative stress, and immunity in liver and differences in glucose and fatty acid pathways in muscle. The current work indicates that unique differences in liver and muscle post-absorptive nutrient metabolism contribute to sources of variation in feed efficiency and that differences in amino acid and fatty acid oxidation, cell cycling, and immune function should be further examined.

Identification and characterization of hypothalamic circular RNAs associated with bovine residual feed intake.

Residual feed intake (RFI) is crucial economic indicator used for calculating the feed efficiency of growing beef cattle. circRNA plays an important biological role in gene transcriptional regulation, but little is known about its potential functional regulation underlying RFI phenotypic variation. As the core center of regulation of animal feeding, the hypothalamus is closely associated with RFI. Therefore, the present study aimed to identify the key genes and functional pathways contributing to variance in cattle RFI phenotypes using RNA sequencing from hypothalamic tissue samples, in order to gain insight into the potential regulatory role of circRNAs in bovine RFI phenotypic variation. Differentially expressed genes were detected by RNA sequencing for beef cattle in the high and low RFI groups, followed by GO, KEGG enrichment, and circRNA-miRNA co-expression network analysis. A total of 257 circRNAs were differentially expressed between the two groups, with 128 significantly upregulated and 129 significantly downregulated genes in H group compared to L group. Among them, 9 unique circRNAs were present in group L and 4 unique circRNAs were present in group H. GO and KEGG enrichment analysis of the source genes of the differentially expressed circRNAs revealed that they were mainly involved in metabolic processes, such as cellular metabolic processes, cellular macromolecular metabolic processes, and regulatory pathways related to nutrient metabolism, including protein and amino acid metabolism, as well as vitamin metabolism and pancreatic secretion associated with the animal feeding behavior. The circRNAs detected in this study were mostly novel, and have not been investigated directly to be associated with the RFI phenotype. Interestingly, most miRNAs of differentially expressed circRNAs predicted based on the circRNA-miRNA co-expression network analysis by using top 50 differentially expressed circRNAs and 13 unique circRNAs, have been reported to be related to animal RFIs, implying that circRNAs in bovine hypothalamic tissue may regulate phenotypic variation in RFI through miRNAs. The study results illustrate the complex biological functions of the hypothalamus in regulating feed efficiency and showing the potential role of circRNAs in the feeding behavior regulation of livestock, which would contributing to expanding the understanding of circRNA.

Dietary Administration of L-Carnitine During the Fattening Period of Early Feed Restricted Lambs Modifies Ruminal Fermentation but Does Not Improve Feed Efficiency.

Early feed restriction of lambs may program animals to achieve reduced feed efficiency traits as a consequence of permanent mitochondrial dysfunction. The hypothesis at the background of the present study is that dietary administration of L-Carnitine (a compound that promotes the activation and transportation of fatty acids into the mitochondria) during the fattening period of early feed restricted lambs can: (a) improve the biochemical profile of early feed restricted lambs, (b) improve feed efficiency, (c) modulate the ruminal and intestinal microbiota, and (d) induce changes in the gastrointestinal mucosa, including the immune status. Twenty-two newborn male Merino lambs were raised under natural conditions but separated from the dams for 9 h daily to allow feed restriction during the suckling period. At weaning, lambs were assigned to a control group being fed ad libitum a complete pelleted diet during the fattening phase (CTRL, n = 11), whereas the second group (CARN, n = 11) received the same diet supplemented with 3 g of L-Carnitine/kg diet. The results revealed that even though L-Carnitine was absorbed, feed efficiency was not modified by dietary L-Carnitine during the fattening period (residual feed intake, p > 0.05), whereas ruminal fermentation was improved [total short-chain fatty acids (SCFAs), 113 vs. 154 mmol/l; p = 0.036]. Moreover, a trend toward increased concentration of butyrate in the ileal content (0.568 vs. 1.194 mmol/100 ml SCFA; p = 0.074) was observed. Other effects, such as reduced heart weight, lower levels of markers related to muscle metabolism or damage, improved renal function, and increased ureagenesis, were detected in the CARN group. Limited changes in the microbiota were also detected. These findings suggest that L-Carnitine may improve ruminal fermentation parameters and maintain both the balance of gut microbiota and the health of the animals. However, the improved ruminal fermentation and the consequent greater accumulation of intramuscular fat might have hidden the effects caused by the ability of dietary L-Carnitine to increase fatty acid oxidation at the mitochondrial level. This would explain the lack of effects of L-Carnitine supplementation on feed efficiency and points toward the need of testing lower doses, probably in the context of animals being fed in excess non-protein nitrogen.

Rumen biohydrogenation and milk fatty acid profile in dairy ewes divergent for feed efficiency.

A sustainable increase in livestock production would require selection for improved feed efficiency, but the mechanisms underlying this trait and explaining its large individual variation in dairy ruminants remain unclear. This study was conducted in lactating ewes to test the hypothesis that rumen biohydrogenation (BH) would differ between high- and low-efficiency animals, and these differences would be reflected in rumen fatty acid (FA) profile and affect milk FA composition. A second aim was to identify differences in FA that may serve as biomarkers of feed efficiency. Data of daily feed intake and milk yield and composition, as well as body weight, were collected individually over a 3-wk period in 40 ewes. The difference between the mean actual and predicted feed intake (estimated through metabolizable energy requirements for maintenance, production, and body weight change) over the period was used as the feed efficiency index (FEI) to select 8 of the highest feed efficiency (H-FE) and 8 of the lowest feed efficiency (L-FE) animals. In addition, residual feed intake (RFI) was estimated as the residual term from the regression of feed intake on various energy sinks. Rumen and milk FA composition were characterized by using gas chromatography, and results were analyzed using a statistical model that included the fixed effect of the group (H-FE vs. L-FE). The FEI averaged -0.29 ± 0.046 and 0.81 ± 0.084 in H-FE and L-FE, respectively, whereas RFI averaged -0.16 ± 0.084 and 0.18 ± 0.082, respectively. The correlation coefficient between both metrics was 0.69. Feed intake was similar in both groups, but H-FE showed greater milk yield, with increases in lactose content and yield, and in milk protein and fat production. Results from rumen FA profiles included a lower proportion of 18:2n-6, cis-9 18:1, and of several of their BH metabolites, and a greater concentration of 18:0, which may indicate that the apparent BH would be more complete in more efficient sheep. Milk FA analysis suggested that the greater fat yield in the H-FE group was mostly explained by increased de novo FA synthesis, whereas their milk would have lower proportions of cis-9 18:1 and C20 to 22n-6 polyunsaturated FA than L-FE. Stepwise multiple linear regression suggested that milk C20 to 22n-6 PUFA might be convenient biomarkers to discriminate more efficient dairy sheep. Further research is needed to validate these findings (e.g., under different dietary conditions).

Between-cow variation in the components of feed efficiency.

A meta-analysis based on an individual-cow data set was conducted to investigate between-cow variations in the components and measurements of feed efficiency (FE) and to explore the associations among these components. Data were taken from 31 chamber studies, consisting of a total of 841 cow/period observations. The experimental diets were based on grass or corn silages, fresh grass, or a mixture of fresh grass and straw, with cereal grains or by-products as energy supplements, and soybean or canola meal as protein supplements. The average forage-to-concentrate ratio across all diets on a dry matter basis was 56:44. Variance component and repeatability estimates of FE measurements and components were determined using diet, period, and cow within experiment as random effects in mixed procedures of SAS (SAS Institute Inc., Cary, NC). The between-cow coefficient of variation (CV) in gross energy intake (GE; CV = 0.10) and milk energy (El) output as a proportion of GE (El/GE; CV = 0.084) were the largest among all component traits. Similarly, the highest repeatability estimates (≥0.50) were observed for these 2 components. However, the between-cow CV in digestibility (DE/GE), metabolizability [metabolizable energy (ME)/GE], methane yield (CH4E/GE), proportional urinary energy output (UE/GE), and heat production (HP/GE), as well as the efficiency of ME use for lactation (kl), were rather small. The least repeatable component of FE was UE/GE. For FE measurements, the between-cow CV in residual energy-corrected milk (RECM) was larger than for residual feed intake (RFI), suggesting a greater possibility for genetic gain in RECM than in RFI. A high DE/GE was associated with increased CH4E/GE (r = 0.24), HP/GE (r = 0.12), ME/GE (r = 0. 91), energy balance as a proportion of GE (EB/GE; r = 0.35), and kl (r = 0.10). However, no correlation between DE/GE and GE intake or UE/GE was observed. Increased proportional milk energy adjusted to zero energy balance (El(0)/GE) was associated with increases in DE/GE, ME/GE, EB/GE, and kl but decreases in UE/GE, CH4E/GE, and HP/GE, with no effect on GE intake. In conclusion, several mechanisms are involved in the observed differences in FE among dairy cows, and reducing CH4E yield (CH4E/GE) may inadvertently result in reduced GE digestibility. However, the selection of dairy cows with improved energy utilization efficiencies offers an effective approach to lower enteric CH4 emissions.

Relationship between performance, metabolic profile, and feed efficiency of lactating beef cows.

Twenty-seven Nellore cow-calf pairs were submitted for feed efficiency testing. The animals were weighed every 21 ± 5 days to obtain metabolic body weight (BW0.75) and average daily gain (ADG). Subcutaneous fat thickness (SFT; at 20, 83, 146, and 176 days post-calving); milk yield and components (63, 85, and 151 days); levels of glucose, cholesterol, triglycerides, ß-hydroxybutyrate, albumin, urea, creatinine, calcium, phosphorus, magnesium, insulin, and cortisol (15, 41, 62, and 124 days); and ingestive behavior were evaluated. Residual feed intake was calculated for the first stage (RFI1; 21 to 100 days post-calving) and the second stage of lactation (RFI2; 100 to 188 days post-calving), and the cows were classified based on RFI1 as most efficient (RFI1 < 0) and least efficient (RFI1 > 0). Negative RFI1 cows consumed 1.3 kg/day of dry matter, or 9.77%, less than positive RFI1 cows. Most- and least-efficient cows did not differ in terms of subcutaneous fat thickness traits and milk yield or energy-corrected milk (ECM). Glucose (P = 0.0785), triglycerides (P = 0.0795), and phosphorus (P = 0.0597) concentrations were higher in the first stage of lactation in most-efficient cows. Maternal characteristics such as calf weight at birth and at 205 days and ADG were similar in most- and least-efficient cows. The most-efficient cows are more economic as they consume less feed for the same level of production.

Feed-efficient pigs exhibit molecular patterns allowing a timely circulation of hormones and nutrients.

Feed efficiency (FE) is a measure of the rate between feed intake and body weight gain and is subject to constant progress in pigs, based on extensive performance tests and analyses of physiological parameters. However, endocrine regulatory circuits that comprise the sensation and perception of intrinsic requirements and appropriate systemic responses have not yet been fully elucidated. It is hypothesized that the gut-brain axis, which is a network of hierarchical anterior regulatory tissues, contributes largely to variations in FE. Therefore, full-sib pigs with extreme residual feed intake values were assigned to experimental groups of high and low FE. Relevant hormones, minerals, and metabolites including fatty acid profiles were analyzed in serum to assess postprandial conditions. Transcriptome profiles were deduced from intestinal (duodenum, jejunum, ileum) and neuroendocrine tissues (hypothalamus). Serum analyses of feed-efficient animals showed an increased content of the incretin GIP, calcium, magnesium, ß-hydroxybutyric acid, and fat compared with low-FE pigs. Complementary expression profiles in intestinal tissues indicate a modulated permeability and host-microbe interaction in FE-divergent pigs. Transcriptomic analyses of the hypothalamus showed that differences between the FE groups in appetite and satiety regulation are less pronounced. However, hypothalamic abundance of transcripts like ADCY7, LHCGR, and SLC2A7 and molecular signatures in local and systemic tissue sites indicate that increased allocation and circulation of energy equivalents, minerals, and hormones are promoted in feed-efficient animals. Overall, patterns of gastrointestinal hormones and gene expression profiles identified host-microbiota interaction, intestinal permeability, feed intake regulation, and energy expenditure as potential mechanisms affecting FE in pigs.

Responses of dairy cows with divergent residual feed intake as calves to metabolic challenges during midlactation and the nonlactating period.

Residual feed intake (RFI) is defined as the difference between the actual and expected feed intake required to support animal maintenance and growth. Thus, a cow with a low RFI can obtain nutrients for maintenance and growth from a reduced amount of feed compared with a cow with a high RFI. Variation in RFI is underpinned by a combination of factors, including genetics, metabolism, thermoregulation and body composition; hypothalamic-pituitary-adrenal (HPA) axis responsiveness is also a possible contributor. Responses to 3 metabolic challenges were measured in lactating and nonlactating dairy cattle. Sixteen Holstein Friesian cows with phenotypic RFI measurements that were obtained during the growth period (188-220 d old) were grouped as either low-calfhood RFI (n = 8) or high-calfhood RFI (n = 8). An ACTH (2 µg/kg of body weight), insulin (0.12 U/kg), and epinephrine (a low dose of 0.1 µg/kg and a high dose of 1.6 µg/kg of epinephrine) challenge were each conducted during both midlactation (122 ± 23.4 d in milk) and the nonlactating period (dry period; approximately 38 d after cessation of milking). Cows were housed in metabolism stalls for the challenges and were fed a diet of alfalfa cubes ad libitum for at least 10 d before the experiment (lactating cows also were offered a total of 6 kg of dry matter/d of crushed wheat grain plus minerals fed as 3 kg of dry matter at each milking) and were fasted for 12 h before the challenges. The efficiency of conversion of feed into milk (the ratio of feed consumed to milk produced over the 7 d before the experiment) during midlactation was better (lower) in low-calfhood RFI cows, although dry matter intake did not differ between RFI groups. Low-calfhood RFI cows exhibited a lower plasma cortisol response to the ACTH challenge than high-calfhood RFI cows, particularly in midlactation (-15%). The low-calfhood RFI cows had a greater plasma insulin-like growth factor-1 response to the insulin challenge and plasma fatty acid response to epinephrine compared with the high-calfhood RFI cows. These data suggest that high-calfhood RFI cows exhibit a more responsive HPA axis. As divergence in RFI measured during growth is retained (although reduced) during lactation, it is possible that energy is used to respond to HPA axis activation at the expense of production in high-calfhood RFI dairy cattle during lactation and contributes to a decrease in overall feed use efficiency.

Association analysis between feed efficiency studies and expression of hypothalamic neuropeptide genes in laying ducks.

Residual feed intake (RFI) is now considered a more reasonable metric to evaluate animal feed efficiency. In this study, the correlation between RFI and other feed efficiency traits was investigated and gene expression within the hypothalamus was determined in low RFI (LRFI) and high RFI (HRFI) ducks. Further, several hypothalamic neuropeptide genes were measured using quantitative real-time PCR. The mean feed intake value was 160 g/day, whereas the egg mass laid (EML) and body weight were approximately 62.4 g/day and 1.46 kg respectively. Estimates for heritability of RFI, feed conversion ratio (FCR) and feed intake were 0.26, 0.18 and 0.23 respectively. RFI is phenotypically positively correlated with feed intake and FCR (P < 0.01). The expression of neuropeptide Y (NPY) and neuropeptide Y receptor Y5 (NPY5R) mRNA was higher in HRFI ducks compared with LRFI ducks (P < 0.05), whereas that of proopiomelanocortin (POMC), melanocortin 4 receptor (MC4R) and cholecystokinin (CCK) was lower (P < 0.05). The mRNA expression of gonadotropin-releasing hormone 1 (luteinizing-releasing hormone) (GNRH1) and prolactin receptor (PRLR) was unchanged between LRFI and HRFI ducks. The results indicate that selection for LRFI could reduce feed intake without significant changes in EML, whereas selection on FCR will increase EML.

Biological implications of longevity in dairy cows: 2. Changes in methane emissions and efficiency with age.

Previous studies indicated that absolute CH4 emissions and CH4 yield might increase and that milk production efficiency might decrease with age in cattle. Both would make strategies to increase longevity in dairy cattle less attractive. These aspects were experimentally determined in Brown Swiss cattle distributed continuously across a large age range. Thirty lactating dairy cows (876-3,648 d of age) received diets consisting of hay, corn silage, and grass pellets supplemented with 0 or 5kg of concentrate per day. Twelve heifers (199-778 d of age) received hay only. Cows and heifers were members of herds subjected to the 2 different feeding regimens (with or without concentrate) for the past 10 yr. Methane emissions were measured individually for 2 d in open-circuit respiration chambers, followed by quantifying individual feed intake and milk yield over 8 d. Additional data on digestibility, rumination time, and passage time of feed of all experimental animals were available. Regression analyses were applied to evaluate effects of age and feeding regimen. Body weight, milk yield, and the hay proportion of forage dry matter intake were considered as covariates. Methane emissions per unit of intake, body weight, and milk yield were significantly related to age. Their development in the cows with age was characterized by an increase to maximum at around 2,000 d of age, followed by a decline. This response was not accompanied by corresponding age-related changes in intake, chewing activity, digesta passage time, and digestibility of organic matter, which would have explained shifts in CH4. However, fiber digestibility showed a similar change with age as methane emissions, resulting in quite stable methane emissions per unit of digestible fiber. As expected, methane emissions intensity per unit of milk produced was greater by 8% without concentrate than with concentrate, but no difference was noted in the response to age when the animals were subjected to different feeding regimens. The efficiency of milk production was only marginally influenced by age and diet, and no different response was observed for age in the 2 dietary regimens. In conclusion, life cycle analyses of milk production systems focusing on longevity should consider changing methane yields with age in addition to the variation in environmental costs for replacements of culled cows.

Effect of inflammation stimulation on energy and nutrient utilization in piglets selected for low and high residual feed intake.

Selection of animals for improved feed efficiency can affect sustainability of animal production because the most efficient animals may face difficulties coping with challenges. The objective of this study was to determine the effects of an inflammatory challenge (using an intravenous injection of complete Freund's adjuvant - CFA) in piglets from two lines of pigs divergently selected during the fattening period for a low (RFI-) or a high (RFI+) residual feed intake (RFI; difference between actual feed intake and theoretical feed requirements). Nitrogen and energy balances (including heat production - HP - and its components: activity-related HP - AHP, thermic effect of feeding, and resting HP) were measured individually in thirteen 20-kg BW castrated male piglets (six and seven from RFI+ and RFI- line, respectively) fed at the same level (1.72 MJ ME/kg BW0.60 per day) from 3 days before to 3 days after CFA injection. Dynamics of dietary U-13C-glucose oxidation were estimated from measurements of 13CO2 production on the day before and 3 days after the CFA injection. Oxidation of dietary nutrients and lipogenesis were calculated based on HP and O2 consumption and CO2 production. The data were analyzed as repeated measurements within piglets in a mixed model. Before CFA injection, RFI- piglets had a lower resting energy expenditure than RFI+ piglets, which tended to increase energy retention because of a higher energy retention as fat. The CFA injection did not affect feed intake from the day following CFA injection onwards but it increased energy retention (P=0.04). Time to recover 50% of 13C from dietary glucose as expired 13CO2 was higher in RFI+ piglets before inducing inflammation but decreased after to the level of RFI- piglets (P<0.01). Oxidation of U-13C-glucose tended to slightly increased in RFI- piglets and to decreased in RFI+ piglets (P=0.10) because of CFA. Additionally, RFI- piglets had a lower respiratory quotient during the 1st day following the CFA injection whereas RFI+ piglets tended to have a higher respiratory quotient. In conclusion, selection for RFI during the fattening period also affected the energy metabolism of pigs during earlier stages of growth. The effects of CFA injection were moderated in both lines but the most efficient animals (RFI-) exhibited a marked re-orientation of nutrients only during the 1st day after CFA, and seemed to recover thereafter, whereas the less efficient piglets expressed a more prolonged alteration of their metabolism.

Inclusion of anti-phospholipase A2 antibody to backgrounding diets on performance, feed efficiency, in vitro fermentation, and the acute-phase response of growing beef calves.

In Exp. 1, individual performance and daily DMI was measured on 70 crossbred weaned calves during a 70-d period using a GrowSafe system (GrowSafe Systems Ltd., Airdrie, AB, Canada) at the University of Florida North Florida Research and Education Center Feed Efficiency Facility (FEF). Calves were fed a low-concentrate (LC) growing diet, blocked by weight and sex, and then randomly assigned to pens to receive either no additional supplement (CON; n = 35) or receive a supplement of anti-phospholipase A2 antibody (aPLA2) at an inclusion rate of 0.6% of the diet DM (n = 35). After the 70-d feed efficiency (FE) trial (Phase 1), calves were loaded into a commercial livestock trailer and were driven for approximately 1,600 km during 24 h. Upon return to the FEF (Phase 2), calves were relocated to the same pens and groups and received the same diets and treatments for 28 d. Blood samples from each calf were collected on d 0, 1, 3, 5, 7, 14, 21, and 28 relative to initiation of transportation and were analyzed for determination of concentrations of plasma ceruloplasmin and haptoglobin. In Phase 1, initial BW (242.0 ± 3.7 kg; P = 0.92), BW at d 70 (313.0 ± 4.1 kg; P = 0.79), and ADG (1.01 ± 0.02 kg; P = 0.95) were similar between treatments. However, daily DMI was greater (P = 0.01) for CON (9.18 ± 0.15 kg) than aPLA2 (8.53 ± 0.15 kg). In addition, residual feed intake was greater (P = 0.002) for CON (0.389 ± 0.110 kg/d) than aPLA2 calves (-0.272 ± 0.110 kg/d). In Phase 2, after transportation, there were no differences between treatments on BW loss due to transportation shrink (26.0 ± 0.6 kg; P = 0.86), BW at d 28 (339.0 ± 4.1 kg; P = 0.72), ADG (1.28 ± 0.03 kg/d; P = 0.72), G:F (0.164 ± 0.004; P = 0.83), and concentrations of plasma haptoglobin (0.08 ± 0.02 mg/mL; P = 0.41). However, concentrations of plasma ceruloplasmin were greater (P < 0.001) for CON calves (14.3 ± 0.3 mg/dL) compared to aPLA2 calves (13.0 ± 0.3 mg/dL). In Exp. 2, the effects of aPLA2 inclusion on LC and high-concentrate (HC) substrates on in vitro fermentation parameters were assessed. Addition of aPLA2 had no effects on in vitro fermentation parameters of LC and HC substrates. In conclusion, supplementation of aPLA2 improved FE of growing beef calves when fed LC diets in Phase 1 and addition of aPLA2 had no effect on fermentation parameters of LC and HC substrates. In addition, calves supplemented with aPLA2 had reduced concentrations of plasma ceruloplasmin after 24 h of transportation.