Expression and secretion of SPARC, FGF-21 and DCN in bovine muscle cells: Effects of age and differentiation.

Skeletal muscle growth is an economically important trait in the cattle industry. Secreted muscle-derived proteins, referred to as myokines, have important roles in regulating the growth, metabolism, and health of skeletal muscle in human and biomedical research models. Accumulating evidence supports the importance of myokines in skeletal muscle and whole-body health, though little is known about the potential presence and functional significance of these proteins in cattle. This study evaluates and confirms that secreted proteins acidic and rich in cysteine (SPARC), fibroblast growth factor 21 (FGF-21), myostatin (MSTN), and decorin (DCN) are expressed and SPARC, FGF-21, and DCN are secreted by primary bovine satellite cells from 3- (BSC3; n = 3) and 11- (BSC11; n = 3) month -old commercial angus steers. Cells were cultured and collected at zero, 12, 24, and 48 hours to characterize temporal expression and secretion from undifferentiated and differentiated cells. The expression of SPARC was higher in the undifferentiated (p = 0.04) and differentiated (p = 0.07) BSC11 than BSC3. The same was observed with protein secretion from undifferentiated (p <0.0001) BSC11 compared to BSC3. Protein secretion of FGF-21 was higher in undifferentiated BSC11 (p < 0.0001) vs. BSC3. DCN expression was higher in differentiated BSC11 (p = 0.006) vs. BSC3. Comparing undifferentiated vs. differentiated BSC, MSTN expression was higher in differentiated BSC3 (p ≤ 0.001) for 0, 12, and 24 hours and in BSC11 (p ≤ 0.03) for 0, 12, 24, and 48 hours. There is also a change over time for SPARC expression (p ≤ 0.03) in undifferentiated and differentiated BSC and protein secretion (p < 0.0001) in undifferentiated BSC, as well as FGF-21 expression (p = 0.007) in differentiated BSC. This study confirms SPARC, FGF-21, and DCN are secreted, and SPARC, FGF-21, MSTN, and DCN are expressed in primary bovine muscle cells with age and temporal differences.

Myokines Produced by Cultured Bovine Satellite Cells Harvested from 3- and 11-Month-Old Angus Steers.

The myokines interleukin 6 (IL-6), interleukin 15 (IL-15), myonectin (CTRP15), fibronectin type III domain containing protein 5/irisin (FNDC5), and brain-derived neurotrophic factor (BDNF) are associated with skeletal muscle cell proliferation, differentiation, and muscle hypertrophy in biomedical model species. This study evaluated whether these myokines are produced by cultured bovine satellite cells (BSCs) harvested from 3- and 11-month-old commercial black Angus steers and if the expression and secretion of these targets change across 0, 12, 24, and 48 h in vitro. IL-6, IL-15, FNDC5, and BDNF expression were greater (p ≤ 0.05) in the differentiated vs. undifferentiated BSCs at 0, 12, 24, and 48 h. CTRP15 expression was greater (p ≤ 0.03) in the undifferentiated vs. differentiated BSCs at 24 and 48 h. IL-6 and CTRP15 protein from culture media were greater (p ≤ 0.04) in undifferentiated vs. differentiated BSCs at 0, 12, 24, and 48 h. BDNF protein was greater in the media of differentiated vs. undifferentiated BSCs at 0, 12, 24, and 48 h. IL-6, 1L-15, FNDC5, and BDNF are expressed in association with BSC differentiation, and CTRP15 appears to be expressed in association with BSC proliferation. This study also confirms IL-6, IL-15, CTRP15, and BDNF proteins present in media collected from primary cultures of BSCs.

The Effects of Harvest Maturity of Eragrostis tef 'Moxie' Hay and Supplemental Energy Source on Forage Utilization in Beef Heifers.

The phenological stage of maturity of grasses and supplementation program can impact forage utilization in grazing beef cattle. However, the potential interaction between harvest maturity of Eragrostis tef (teff) hay and energy supplement source was yet to be fully evaluated. Therefore, our objective was to determine the effects of harvest maturity of teff hay and supplemental energy sources on nutrient intake, apparent total-tract nutrient digestion, nitrogen (N) utilization, and ruminal fermentation characteristics in beef heifers. A split-plot design with teff hay harvest maturity as the whole plot and supplemental energy source as the subplot was administered in a three-period (21 d), three × three Latin square design. Six crossbred beef heifers (804 ± 53.6 kg of body weight; BW) were allocated to two harvest maturities (early- (EH]) or late-heading (LH)) and to two supplemental energy sources (no supplement (CON), or rolled corn grain or beet pulp pellet fed at 0.5% of BW). Data were analyzed using SAS. There was no harvest maturity × energy supplement interaction. Although harvest maturity had no impact on total dry matter intake (DMI), crude protein (CP) intake was greater (p < 0.01) for EH than LH heifers. Total intakes of dry (DM) and organic matter (OM) were also greater (p < 0.01) for supplemented than CON heifers, whereas acid detergent fiber (ADF) intake was greater for beet pulp heifers compared to heifers fed the CON diet and supplemental corn grain. Harvest maturity had no impact on ruminal pH. However, mean ruminal pH was lower (p = 0.04), duration pH < 6.2, and molar proportions of butyrate and branched-chain fatty acids were greater (p ≤ 0.049) for heifers fed corn grain compared to CON and beet pulp diets. Heifers fed EH hay had greater (p ≤ 0.02) apparent total-tract DM, OM, CP, NDF, and ADF digestibility than heifers fed LH hay. Although there was no supplemental energy effect on microbial nitrogen (N) flow, it was greater (p < 0.01) for EH than LH heifers. Apparent N retention, which did not differ, was negative across all diets. In summary, delaying the harvest of teff hay from the EH to LH stage of maturity compromised nutrient supply, which was not attenuated by feeding supplemental corn grain and beet pulp at 0.5% of diet DM. Because N retention was negative across harvest maturity, there might be a need to provide both energy and protein supplements to improve growth performance when feeding teff hay to beef cattle.

Inclusion of Grape Pomace in Finishing Cattle Diets: Carcass Traits, Meat Quality and Fatty Acid Composition.

Because of its high content of polyphenolic compounds, dietary inclusion of grape pomace (GP) in finishing cattle diet could possibly enhance product quality and the health value of beef lipids. Therefore, the aim of this study was to evaluate the effects of feeding a high amount of grape pomace in finishing cattle diets on carcass traits, product quality, and fatty acid (FA) composition of beef. Jersey × Holstein crosses (n = 24) were fed either a typical finishing diet (CON) or a finishing diet containing 58% grape pomace (DM basis; HGP). Following the feeding period, animals were harvested, and carcass traits measured. Longissimus lumborum (LL) and semimembranosus (SM) muscle were then collected from each carcass for sensory quality evaluation and FA profile analysis. Hot carcass weight, backfat thickness, and preliminary and final yield grades were greater (p ≤ 0.04) for CON than HGP steers. However, there was no diet effect on rib eye area (REA), kidney, pelvic, and heart (KPH) fat, and marbling. Feeding the HGP compared to CON diet reduced lipid oxidation of LL and SM steaks over time; the malondialdehyde (MDA) concentration, which did not differ on d 0 and 2 of 8-d simulated retail display, was lower on d 4, 6 and 8 for HGP than CON steers (treatment × day of simulated display interaction; p < 0.01). Brightness (L* values) and redness (b*) were greater for LL steaks from HGP than CON steers on most days of simulated display (treatment × day of simulated display interaction; p < 0.01). In addition, the LL and SM muscle content of several FA linked to positive health outcomes in humans including 18:2 n-6, 18:2 c9t11, total conjugated linoleic acid (CLA) and total polyunsaturated fatty acid (PUFA) was also greater (p ≤ 0.02) for steers fed the HGP compared to the CON diet. In summary, current findings suggest that although it could possibly limit growth performance, feeding a high amount of grape pomace to finishing cattle could enhance both the sensory quality and the health value of beef lipids, which are key in increasing consumer acceptability of beef.

Increasing dietary proportion of wheat grain in finishing diets containing distillers' grains: impact on nitrogen utilization, ruminal pH, and digestive function.

Because of its high crude protein (CP) content, dietary inclusion of corn dried distillers' grains with solubles (DDGS) in finishing cattle diets can increase the ruminal loss of ammonia-nitrogen (NH3-N), which ends up excreted as urine urea-N (UUN). Increasing dietary fermentable energy supply can enhance ruminal use of N; however, it could also lead to acidotic conditions that compromise digestive function and animal performance. We evaluated the effects of partially replacing dietary corn grain with 20% or 40% (dry matter [DM] basis) wheat grain in finishing diets containing 15% corn DDGS on N utilization, ruminal pH, and digestive function. Nutrient intake and digestion, ruminal fermentation characteristics, microbial protein synthesis, route of N excretion, and blood metabolites were measured. Six ruminally fistulated crossbred beef heifers (initial body weight ± SD; 797 ± 58.8 kg) were used in a replicated 3 × 3 Latin square design with 28-d periods. Dietary treatments were either corn (73% of diet DM; CON), 53:20 corn:wheat blend (20W), or 33:40 corn:wheat blend (40W) as the major fermentable energy source. Dry matter intake (DMI) tended to be lower for heifers fed the 40W than CON and 20W diets. Feeding diets containing wheat grain led to an increase (P = 0.04) in neutral detergent fiber (NDF) intake. However, there was no diet effect (P ≥ 0.60) on apparent total tract DM and NDF digestibility. Feeding wheat grain led to a decrease (P ≤ 0.03) in mean and minimum pH, an increase (P = 0.04) in pH < 5.8 duration, and a tendency for an increase in the area and acidosis index for pH < 5.8 and 5.5. Nitrogen intake, which was lower (P = 0.04) for 40W than 20W heifers did not differ between CON and 20W heifers. There was no diet effect (P = 0.80) on ruminal NH3-N concentration and estimated microbial N flow. However, feeding diets containing wheat grain led to a decrease (P = 0.045) in UUN excretion (% total urine N). Fecal and total N excretion (% of N intake) increased (P < 0.01) following the addition of wheat grain to the diet. Apparent N retention was lower (P = 0.03) for 40W than CON and 20W heifers. In summary, although it led to a desirable decrease in UUN excretion, feeding wheat grain in corn DDGS-containing diets increased acidotic conditions in the rumen, which possibly led to the tendency for a decrease in DMI. The negative apparent N retention at the 40% wheat grain inclusion also suggests a decrease in nutrient supply, which could compromise feedlot performance.

Optimum grape pomace proportion in feedlot cattle diets: ruminal fermentation, total tract nutrient digestibility, nitrogen utilization, and blood metabolites.

Because of its high content of polyphenolic compounds, the dietary inclusion of grape pomace (GP) in ruminant diets can reduce reactive nitrogen (N) and methane emissions and enhance the shelf life and beneficial fatty acids (FAs) content of meat. However, the dietary inclusion of GP beyond a threshold that is still to be determined for feedlot cattle can also compromise nutrient supply and, thus, growth performance. This study investigated the optimum proportion of GP in finishing cattle diets. Nutrient intake and apparent total tract digestion, ruminal pH and fermentation, estimated microbial protein synthesis, route of N excretion, and blood metabolites were measured. Six ruminally fistulated crossbred beef heifers (mean initial body weight ± SD: 714 ± 50.7 kg) were used in a replicated 3 × 3 Latin square with 21-d periods. Dietary treatments were 0%, 15%, and 30% of dietary dry matter (DM) as GP, with diets containing 84%, 69%, and 54% dry-rolled barley grain, respectively. There was a linear increase (P = 0.07) in DM intake and quadratic change (P ≤ 0.01) in neutral detergent fiber (NDF) intake. There was a quadratic change (P ≤ 0.04) in apparent total tract DM, NDF, and crude protein digestibility as dietary GP content increased. However, there were no treatment effects (P ≥ 0.18) on total ruminal short-chain FA concentration and duration and area pH < 6.2, 5.8, and 5.5. Although N intake did not differ (269, 262, 253 g/d; P = 0.33) across dietary treatments, feeding GP led to a tendency for a quadratic change (P ≤ 0.07) in ruminal ammonia-N and plasma urea-N concentrations. Total N excretion also changed (quadratic, P = 0.03) because of changes (quadratic, P = 0.02) in fecal N excretion as urinary excretion of N and urea-N did not differ (P ≥ 0.15) across treatments. Feeding GP led to quadratic changes (P ≤ 0.01) in fecal excretion of fiber-bound N. Microbial N flow and apparent N retention also changed (quadratic, P ≤ 0.04) as dietary GP proportion increased. In conclusion, responses to dietary GP proportion were mostly quadratic with indications that nutrient supply as reflected by changes in apparent total tract nutrient digestibility, microbial N supply, and apparent N retention could be compromised beyond a 15% dietary inclusion level.

Optimum roughage proportion in barley-based feedlot cattle diets: growth performance, feeding behavior, and carcass traits.

High grain diets are fed to finishing beef cattle to maximize animal performance in a cost-effective manner. However, a small amount of roughage is incorporated in finishing diets to help prevent ruminal acidosis, although few studies have examined optimum roughage inclusion level in barley-based diets. The objective of the study was to evaluate the effects of roughage proportion in barley-based finishing diets on growth performance, feeding behavior, and carcass traits of feedlot cattle. Crossbred beef steers (n = 160; mean body weight ± SD, 349.7 ± 21.4 kg) were allocated to 20 pens that were assigned randomly to four dietary treatments (five pens of eight steers per treatment). The treatment diets contained barley silage at 0%, 4%, 8%, and 12% of dietary dry matter (DM). The remainder of the diets (DM basis) consisted of 80%, 76%, 72%, and 68% barley grain, respectively, 15% corn dried distiller's grains, 5% mineral and vitamin supplement, and 32 mg monensin/kg diet DM. The diets were fed as total mixed rations for ad libitum intake (minimum of 5% refusal) once per day. Cattle were weighed on 2 consecutive days at the start and end of the experiment and on 1 d every 3 wk throughout the experiment (124 d). Two pens for each treatment group were equipped with an electronic feeding system (GrowSafe Systems Ltd., Calgary, Alberta) to monitor feed intake and feeding behavior of individual cattle. The data for dry matter intake (DMI), average daily gain (ADG), gain:feed (G:F) ratio, and carcass traits were analyzed as a completely randomized design with fixed effect of barley silage proportion and pen replicate as experimental unit. Feeding behavior data were analyzed similarly, but with animal as experimental unit. Averaged over the study, DMI increased linearly (11.1, 11.3, 11.7, 11.8 kg/d; P = 0.001) as barley silage proportion increased from 0%, 4%, 8%, and 12% of DM, but ADG was not affected (carcass-adjusted,1.90, 1.85, 1.87, 1.89 kg/d; P ≥ 0.30). Consequently, G:F ratio decreased linearly (carcass-adjusted, 168.9, 163.8, 158.5, 160.6 g/kg DMI; P = 0.023). When averaged over the study, proportion of barley silage in the diet had no linear or quadratic effects (P > 0.10) on meal frequency, duration of meals, intermeal duration, or meal size, but eating rate decreased linearly with increasing silage proportion (P = 0.008). There was no diet effect on liver abscesses (P ≥ 0.92), and effects on carcass characteristics were minor or nonexistent. We conclude that increasing the proportion of barley silage in a feedlot finishing diet at the expense of barley grain to minimize the incidence of ruminal acidosis may decrease feed conversion efficiency.

Optimum roughage proportion in barley-based feedlot cattle diets: total tract nutrient digestibility, rumination, ruminal acidosis, short-chain fatty absorption, and gastrointestinal tract barrier function.

Cattle need physically effective fiber to promote rumination and maintain rumen health, but economics favor the use of low-roughage feedlot diets. The study investigated the optimum barley silage proportion in barley-based finishing diets. Apparent total-tract digestibility (4-d total fecal collection), chewing behavior (6-d video recording), ruminal pH (6-d indwelling pH recording), and fermentation (1 day, sampling 0, 3, 6, 12, and 18 h postfeeding), short-chain fatty acid (SCFA) absorption (washed reticulo-rumen technique), gastrointestinal tract barrier function (marker infusion), and blood variables (catheters) were measured. Eight ruminally fistulated crossbred beef heifers (653 ± 44.2 kg; mean starting body weight [BW] ± SD) were used in a replicated 4 × 4 Latin square design with 28-d periods. Dietary treatments were 0%, 4%, 8%, and 12% of dietary dry matter (DM) as barley silage, with diets containing 80%, 76%, 72%, and 68% barley grain, respectively. Increasing silage proportion decreased dietary starch content from 49.0% to 43.1% DM, while neutral detergent content increased from 22.7% to 25.1% DM. Silage proportion had no effect on DM intake, but apparent DM digestibility decreased quadratically (86.0%, 82.1%, 81.1%, 79.5% for the four diets, respectively; P < 0.001). Although, silage proportion had no effect on eating activity, rumination time increased quadratically (246, 289, 302, 316 min/d; P = 0.04). Increased silage proportion increased minimum (5.07, 5.27, 5.29, 5.41; quadratic, P = 0.011) and mean (5.61, 5.87, 5.93, 5.95; quadratic, P = 0.007) ruminal pH, and there was a quadratic (P ≤ 0.047) decrease in duration and area under the pH acidosis threshold curves of 5.8, 5.5, and 5.2. Although increasing silage proportion decreased ruminal acidosis, it was not completely eliminated even with a diet containing 12% silage DM. SCFA concentration in ruminal fluid was not affected by diet, but silage proportion quadratically (P ≤ 0.088) increased ruminal acetate:propionate. There was no effect of diet on absolute or fractional rates of absorption of acetate, propionate, butyrate or total SCFA, and no effect on gastrointestinal barrier function or blood measurements. In conclusion, responses to roughage level were mostly quadratic with greatest improvements in acidosis variables between 0% and 4% barley silage, with incremental improvements with further increases in silage levels. The study showed a trade-off between maximizing digestibility and energy intake to promote animal performance and minimizing the risk of acidosis.

Effects of maturity at harvest on the nutritive value and ruminal digestion of Eragrostis tef (cv. Moxie) when fed to beef cattle.

Teff (Eragrostis tef cv. Moxie), a warm-season annual grass, could be an excellent forage for beef cattle. However, there is limited information on its nutritive value to cattle when harvested at different stages of maturity. Thus, the objective of this research was to determine the effect of feeding teff hay harvested at the boot (BT), early-heading (EH), or late-heading (LH) stages of maturity on nutrient intake, ruminal fermentation characteristics, omasal nutrient flow, and N utilization in beef cattle. Six ruminally cannulated beef heifers (mean initial BW ± SD, 476 ± 32.6) were used in a replicated 3 × 3 Latin square design with 28-d periods (18 d for adaptation and 10 d for measurements). Dry matter intake was measured daily. Indwelling pH loggers were used to measure ruminal pH from days 21 to 28. Ruminal fluid and omasal digesta were collected from days 26 to 28 to determine fermentation characteristics and omasal nutrient flow. Fecal and urine samples to quantify N excretion were also collected (days 26 to 28). Blood samples for plasma urea-N (PUN) determination were collected 3 h post-feeding on day 28. There were no changes (P > 0.28) in the ADF or NDF content of teff with advancing maturity, but indigestible NDF increased (P < 0.01) with increasing maturity. Maturity had no effect (P ≥ 0.14) on DMI, and ruminal total short-chain fatty acid (SCFA) concentration, pH, digestibility, and outflow of DM, OM, NDF, ADF, and CP. However, the CP content of BT hay was greater (P < 0.01) than for EH and LH hay (18.1, 14.1, and 11.5%, respectively, DM basis), and this resulted in the higher CP intake (P < 0.01) for heifers fed the BT than the EH and LH hay. Consequently, ruminal ammonia-N (NH3-N) concentration was greater (P < 0.01) for heifers fed BT than EH and LH hay, thereby possibly explaining the tendency for a decrease (P = 0.08) in PUN concentration, and a decrease (P < 0.01) in the excretion of total N, urine N, and urea-N (UUN) with advancing maturity. However, fecal N excretion (g/d) did not differ (P = 0.76). In conclusion, despite a decrease in CP intake and ruminal NH3-N concentration, feeding beef heifers EH and LH compared to BT teff hay did not compromise ruminal digestion and outflow of DM, OM, NDF, ADF, and CP, and microbial protein synthesis. Advancing maturity in teff hay also resulted in a decrease in the excretion of total N and urine N and UUN when fed to cattle.