Time course of changes in growth factor mRNA levels in muscle of steroid-implanted and nonimplanted steers

We used a muscle biopsy technique in conjunction with real-time PCR analysis to examine the time course of changes in muscle IGF-I, IGFBP-3, myostatin, and hepatocyte growth factor (HGF) mRNA in the longissimus muscles of Revalor-S-implanted and nonimplanted steers on d 0, 7, 12, and 26 after implantation (nine steers/treatment group). Administration of a Revalor-S implant increased (P < 0.01) ADG and improved (P < 0.05) feed efficiency, 36 and 34%, respectively, compared with steers that received no implant during the 26-d trial. Daily dry matter intake did not differ (P > 0.15) between nonimplanted and implanted steers. Steers receiving the Revalor-S implant had increased (P < 0.001) circulating IGF-I concentrations compared with nonimplanted steers. The longissimus muscles of steers receiving the Revalor-S implant contained increased (P < 0.001) IGF-I mRNA levels compared with longissimus muscles of nonimplanted steers over the 26-d duration of the study. Longissimus muscle IGF-I mRNA levels in implanted steers were increased (P < 0.003) relative to d-0 concentrations on d 7 and 12 (101% and 128%, respectively), and byd 26, longissimus muscle mRNA levels were more than three times (P < 0.0001) those in the longissimus muscles of the same steers on d 0. There was no treatment effect on the level of IGFBP-3, myostatin, or HGF mRNA in the longissimus muscle at any time point; however, levels of IGFBP-3, myostatin, and HGF mRNA increased with time on feed. Based on current and previous studies, we hypothesize that the increased IGF-I level in muscle of implanted steers by d 7 of implantation stimulates satellite cell proliferation and maintains a high number of proliferating satellite cells at a point in the growth curve where satellite cell numbers and activity are normally dropping off. This would prolong the period of rapid muscle growth, resulting in the observed increased rate and efficiency of muscle deposition in implanted steers.     

Effects of flax supplementation and a combined trenbolone acetate and estradiol implant on circulating insulin-like growth factor-I and muscle insulin-like growth factor-I messenger RNA levels in beef cattle

We evaluated effects of a 5% (dry matter basis) ground flaxseed supplement (flax) and a trenbolone acetate and estradiol-17beta implant, Revalor-S, on circulating IGF-I and muscle IGF-I messenger RNA (mRNA). Sixteen crossbred yearling steers (initial BW = 397 kg) were assigned randomly to one of four treatments: 1) flax/implant; 2) nonflax/implant; 3) flax/nonimplant; and 4) nonflax/nonimplant. Serum was harvested from blood collected on d 0 (before implant or flax addition), 14, and 28, and used in subsequent analyses of circulating IGF-I. Biopsy samples (0.5 g) were obtained from the longissimus muscle on d 0, 14, and 28. Total RNA was isolated from the muscle samples, and real-time quantitative-PCR was used to assess relative differences in IGF-I mRNA. Flax supplementation had no effect (P > 0.10) on circulating IGF-I concentrations. Following implantation, sera from implanted steers had 52 and 84% greater (P < 0.05) IGF-I concentrations than sera from nonimplanted steers on d 14 and 28, respectively. On d 28, local muscle IGF-I mRNA levels increased 2.4-fold (P < 0.01) in biopsy samples obtained from implanted compared with nonimplanted steers. Muscle biopsy samples from nonflax cattle had 4.4-fold higher (P < 0.01) levels of IGF-I mRNA than those from flax cattle on d 28. To determine whether a component of flax, alpha-linolenic acid (alphaLA), was directly responsible for IGF-I mRNA down-regulation, we incubated primary cultures of bovine satellite cells, from implanted and nonimplanted steers, in two concentrations of alphaLA (10 nM and 1 microM). An implant x dose interaction (P < 0.05) was observed for IGF-I mRNA concentrations in bovine satellite cells cultured for 72 h with alphaLA. Satellite cells from nonimplanted steers had similar (P > 0.10) IGF-I mRNA concentration regardless of the level of alphaLA exposure; however, satellite cells from implanted steers exposed to 10 nM and 1 microM alphaLA had 2.5- and 2.0-fold greater IGF-I mRNA levels, respectively, than cells from implanted steers that were not exposed to alphaLA (P < 0.05). Administration of a Revalor-S implant increased circulating IGF-I and local muscle IGF-I mRNA concentrations in finishing cattle. However, muscle IGF-I mRNA levels were decreased by flax supplementation. Muscle cell culture experiments suggested that alphaLA was not responsible for the IGF-I mRNA down-regulation.     

Implant strategies during feeding: impact on carcass grades and consumer acceptability

Anabolic growth promotants influence beef grade factors and Warner-Bratzler shear force of steaks. No study has assessed the consumer acceptability of beef derived from implanted cattle. This study determined beef carcass grades and consumer acceptability for cooked beef from unimplanted (control) cattle and from cattle implanted with one of seven different implant strategies (initial implant/implant at 59 d = Encore & Component T-S/no implant, Ralgro/Synovex Plus, Ralgro/Revalor-S, Revalor-S/Revalor-S, Revalor-S/no implant, no implant/Synovex Plus, and Synovex Plus/no implant). British crossbred steers (n = 448) were allocated randomly into one of eight pens for each of the control and seven treatment groups. Carcass quality and yield grade (n = 403) and Warner-Bratzler shear force (n = 298) data were collected by trained personnel. Twenty steaks per control or treatment group were selected randomly for use in consumer sensory evaluation. Steaks were evaluated by consumers for overall like, tenderness like, tenderness level, flavor like, flavor intensity, and juiciness level using 9-point, end-anchored hedonic scales. Control carcasses had smaller (P < .05) longissimus muscle areas than carcasses in all treatment groups except those receiving Encore & Component T S/no implant, Ralgro/Synovex Plus, or Revalor S/no implant. Control carcasses had higher (P < .05) marbling scores than carcasses in all treatment groups except those receiving Ralgro/Revalor-S or Encore & Component T-S/no implant. Steaks from control steers had lower (P < .05) Warner-Bratzler shear force values than steaks from steers given Revalor-S/no implant. Consumer ratings for tenderness like and tenderness level were influenced (P < .05) by implant strategy. Effects of implant strategy on overall like, flavor like, and flavor intensity approached significance (P = .07 to .09). Consumers rated steaks from unimplanted steers as more tender (tenderness level; P < .05) than steaks from all treatment groups except that involving Encore & Component T-S/no implant. Consumers rated steaks from unimplanted steers as more desirable (P < .05) for tenderness like than steaks from all treatments except those involving Encore & Component T-S/no implant or Revalor-S/no implant. Although use of implants in this study resulted in heavier hot carcass weights and larger ribeyes, some of the implant strategies reduced consumer preference of tenderness of steaks.     

Effect of an accelerated finishing program on performance,carcass characteristics,and circulating insulin-like growth factor I concentration of early-weaned bulls and steers

Sixty-three Angus x Simmental calves were allotted to a bull or a steer group based on sire, birth date, and birth weight to determine effects of castration status on performance, carcass characteristics, and circulating insulin-like growth factor I (IGF-I) concentrations in early-weaned cattle. At 75 d of age, calves in the steer group were castrated. Calves were not creep-fed prior to weaning. All calves were weaned and weighed at an average age of 115 d and transported by truck to the OARDC feedlot in Wooster, OH. Performance and carcass characteristics were measured in three phases. Phase 1 was from 115 to 200 d of age, phase 2 was from 201 to 277 d of age, and phase 3 was from 278 d of age to slaughter. Before implantation, four bulls and four steers were selected for serial slaughter and carcass evaluation. Steers were implanted with Synovex-C at 130 d of age and with Revalor-S at 200 and 277 d of age. Serum samples were collected from all calves on the day of implantation, 28 and 42 d after implantation, and at slaughter and analyzed for circulating IGF-I concentration. Bulls gained 9.7% faster (1.75 vs 1.60 kg/d; P < 0.01), consumed 25 kg more DM (521 vs 496 kg; P = 0.11), and were 3.3% more efficient (282 vs 273 g/kg, P < 0.10) than steers in phase 1. However, steers gained 10.5% faster (1.62 vs 1.46 kg/d; P < 0.02), consumed similar amounts of DM, and were 6.5% more efficient than bulls (214 vs 201 g/kg; P < 0.06) in phase 2. Overall gains and efficiency were similar between bulls and steers; however, bulls consumed 140 kg more DM (P < 0.05), were 27 kg heavier (P < 0.05), and had to stay in the feedlot 18 more days (P < 0.05) than steers to achieve a similar amount of fat thickness. Implanted steers had greater concentrations of circulating IGF-I than bulls (P < 0.01), and the pattern of IGF-I concentration over time was affected by castration status (castration status x time interaction; P < 0.01). Synovex-C had a lower impact on circulating IGF-I concentration (implant effect, P < 0.01) than either Revalor-S implant. Eighty-five percent of both bulls and steers had marbling scores sufficient to grade low Choice or better. Bulls achieved their target fat thickness later, increased muscle growth, and deposited fat more favorably than steers, possibly due to a gradual increase in IGF-I concentration as the testicles grew rather than the large fluctuations in IGF-I concentration observed in steers following implantation.     

Response to ractopamine-hydrogen chloride is similar in yearling steers across days on feed

Yearling steers (n = 2,552; 314 kg of initial BW) were used to evaluate the effects of ractopamine-HCl (RAC) and days on feed on performance, carcass characteristics, and skeletal muscle gene expression in finishing steers. Treatment groups included serial slaughter dates of 150, 171, or 192 d on feed. Within each slaughter date, steers either received RAC (200 mg/steer) daily for the final 28 d or were not fed RAC. All steers were initially implanted with Revalor-IS and were reimplanted with Revalor-S after 75 d on feed. At slaughter, muscle samples from the semimembranosus were collected for mRNA analysis of the beta-adrenergic receptors (beta-AR). Ractopamine administration increased (P < 0.05) ADG, G:F, and HCW and increased (P = 0.08) LM area. Ractopamine did not affect the dressing percentage, USDA yield grade, or quality grade (P > 0.3). There was no change in overall feed intake across the entire feeding period; however, feed intake was increased during the 28-d period during which the steers were fed RAC (P < or = 0.05). Greater days on feed decreased (P < 0.05) ADG, G:F, DMI, and the number of yield grade 1 and 2 carcasses. Also, greater days on feed increased (P < 0.05) HCW, dressing percentage, and the number of prime and choice carcasses, as well as the number of yield grade 4 and 5 carcasses. Increasing days on feed decreased (P < 0.05) the abundance of beta(1)-AR and beta(3)-AR mRNA and increased (P < 0.05) the abundance of beta(2)-AR mRNA in skeletal muscle samples obtained at slaughter. Ractopamine had no effect (P > 0.10) on the abundance of beta(1)-AR or beta(3)-AR mRNA, but tended (P = 0.09) to increase beta(2)-AR mRNA. Additional time-course studies with primary muscle cell cultures revealed that advancing time in culture increased (P < 0.001) beta(2)-AR mRNA but had no effect (P > 0.10) on beta(1)-AR or beta(3)-AR mRNA. We conclude that days on feed and RAC are affecting beta-AR mRNA levels, which could, in turn, impact the biological response to RAC feeding in yearling steers.     

Effect of flax supplementation and growth promotants on lipoprotein lipase and glycogenin messenger RNA concentrations in finishing cattle

Lipoprotein lipase (LPL) hydrolyzes triacylglycerols into monoacylglycerol and fatty acids, which are taken up by tissues and used for energy. Glycogenin is the core protein on which glycogen molecules are synthesized. There is one molecule of glycogenin per molecule of glycogen in skeletal muscle; therefore, glycogen storage is limited by the amount of glycogenin present in muscle. The objective of this study was to investigate the effect of feeding flaxseed, a source of PUFA, and administering a growth promoter on steady-state LPL and glycogenin mRNA content of muscle in finishing cattle. Sixteen crossbred steers (initial BW = 397 kg), given ad libitum access to a 92% concentrate diet for 28 d, were used in a four-treatment, 2 x 2 factorial experiment, with flaxseed supplementation (0 or 5% of dietary DM) and implanting (not implanted or implanted with Revalor-S) as the main effects. Muscle biopsies were obtained from the LM at 0, 14, and 28 d, and used to quantify LPL and glycogenin mRNA concentrations using real-time quantitative PCR. Implanting with Revalor-S did not affect LPL (P = 0.13) or glycogenin (P = 0.98) mRNA concentrations. A day x flaxseed interaction (P < 0.001) was observed for both LPL and glycogenin mRNA concentrations. No differences (P > 0.10) were observed between 0 and 5% flaxseed supplemented steers; however, at 28 d, nonflaxseed-fed steers had 4.1- and 5.7-fold increases (P < 0.001) over flaxseed steers for LPL and glycogenin mRNA concentrations, respectively. To further evaluate the effects of alpha-linolenic acid (alpha-LA) on LPL and glycogenin mRNA concentrations, muscle satellite cells were isolated from five finishing steers, and different alpha-LA concentrations were applied in culture. The RNA was isolated from the bovine satellite cells. Addition of alpha-LA numerically increased (P = 0.16) the LPL mRNA concentration 48% at 1 microM alpha-LA compared with the control. The expression of glycogenin was increased (P < 0.05) 50% at 1 microM alpha-LA compared with the control. These results suggest that flaxseed supplementation to finishing steers for 28 d decreased gene expression of both LPL and glycogenin compared with not feeding flaxseed. Alterations in local concentrations of these two proteins could affect the ability of muscle to use fatty acids and glucose for energy, and, ultimately, affect carcass quality.     

Effects of implants on daily gains of steers wintered on dormant native tallgrass prairie, subsequent performance, and carcass characteristics

Fall-weaned crossbred steer calves (n = 300; 184 +/- 2.9 kg) received either no implant (Control) or were implanted with Synovex-C (SC = 10 mg estradiol benzoate + 100 mg progesterone), Synovex-S (SS = 20 mg estradiol benzoate + 200 mg progesterone), or Revalor-G (RG = 8 mg estradiol-17beta + 40 mg trenbolone acetate) to determine the effects of implants on weight gain during winter grazing on dormant tallgrass prairie, subsequent grazing and finishing performance, and carcass characteristics. Steers grazed two dormant tallgrass prairie pastures from October 16, 1996, until March 29, 1997 (164 d), and received 1.36 kg/d of a 25% CP supplement that supplied 100 mg of monensin/steer. Following winter grazing, all steers were implanted with Ralgro (36 mg zeranol) and grazed a common tallgrass prairie pasture until July 17 (110 d). After summer grazing, all steers were implanted with Revalor-S (24 mg estradiol-17beta + 120 mg trenbolone acetate), and winter implant treatment groups were equally allotted to four feedlot pens. Steers were harvested November 17, 1997, after a 123-d finishing period. Daily gains during the winter grazing phase averaged .28, .32, .32, or .35 kg/d, respectively, for Control, SC, SS, or RG steers and were greater (P < .01) for implanted steers than for Controls. Summer daily gains were similar (1.05 +/- .016 kg/d; P > or = .61) for all treatment groups. Feedlot daily gains were also similar (1.67 +/- .034 kg/d; P > or = .21), with implanted steers weighing 14 kg more than Control steers (P = .05) at harvest, despite similar management during summer grazing and feedlot phases. Control steers tended (P = .06) to have lower yield grades. There were no differences (P = .99) in marbling between implanted and nonimplanted steers. Steers implanted during the wintering phase had increased skeletal and overall (P < .01) carcass maturities compared with nonimplanted steers, which resulted in more "B" and "C" maturity carcasses. Because carcass maturity score affects quality grade, the increased maturities of implanted steers resulted in a $9.04 decrease in carcass value/100 kg (P < .01) compared with Controls. The results of this study indicate that growth-promoting implants are efficacious for cattle wintered on dormant native range despite low daily gains. This increased weight is maintained through the summer grazing and feedlot phases; however, the benefit of the increased weight may be offset by decreased carcass quality grade and value due to increased carcass maturity.     

Increased degradation of insulin-like growth factor-I in serum from feed-deprived steers

Severe feed restriction decreases serum insulin-like growth factor I (IGF-I) concentration in animals, and this decrease is thought to be due to reduced IGF-I production in the liver. The objective of this study was to determine whether feed deprivation also increases degradation of serum IGF-I and serum levels of IGF binding protein 3 (IGFBP-3) and acid-labile subunit (ALS), which inhibit IGF-I degradation and increase IGF-I retention in the blood by forming a ternary complex with IGF-I, in cattle. Five steers had free access to pasture, and another five were deprived of feed for 60 h. Serum concentration of IGF-I and liver abundance of IGF-I mRNA at the end of the 60-h period were 50% and 80% lower, respectively, in feed-deprived steers than in fed steers. Less ¹²⁵I-labeled IGF-I remained intact after a 45-h incubation in sera of feed-deprived steers than in sera of fed steers, suggesting that serum IGF-I is more quickly degraded in feed-deprived animals. Serum levels of IGFBP-3 and ALS were decreased by 40% and 30%, respectively, in feed-deprived steers compared with fed steers. These decreases were associated with more than 50% reductions in IGFBP-3 and ALS mRNA in the liver, the major source of serum IGFBP-3 and ALS. Taken together, these results suggest that feed deprivation reduces serum concentration of IGF-I in cattle not only by decreasing IGF-I gene expression in the liver, but also by increasing IGF-I degradation and reducing IGF-I retention in the blood through decreasing IGFBP-3 and ALS production in the liver.     

Effects of repetitive use of hormonal implants on beef carcass quality,tenderness, and consumer ratings of beef palatability

Effects of repetitive use of anabolic implants on beef carcass quality, tenderness, and consumer ratings for palatability were investigated using crossbred steer calves (n = 550). Steers from five ranches were randomly allocated to one of 10 different lifetime implant strategies or to a nonimplanted control group. Cattle were implanted at some or all of five phases of production (branding, weaning, backgrounding, feedlot entry, or reimplant time). Carcasses from the control group had higher (P < 0.05) marbling scores than carcasses from steers in all other treatment groups. Implanting steers at branding, weaning, or backgrounding vs. not implanting steers at these production stages did not affect (P > 0.05) marbling scores. Steers implanted twice during their lifetime produced carcasses with higher (P < 0.05) marbling scores than did steers receiving a total of four or five implants. Steaks obtained from carcasses in the control group had lower (P < 0.05) shear force values and were rated by consumers as more desirable (P < 0.05) for tenderness like/dislike than steaks obtained from carcasses in all other treatment groups. Implanting steers at branding or weaning production stages did not affect (P > 0.05) steak shear force values, consumer ratings for like/dislike of steak tenderness, or percentage of consumers rating overall eating quality of steaks as satisfactory. Implanting steers at backgrounding vs. not implanting steers at this production stage increased (P < 0.05) steak shear force values, but did not influence (P > 0.05) consumer ratings for like/dislike of steak tenderness or percentage of consumers rating overall eating quality of steaks as satisfactory. Steaks from nonimplanted steers were rated as more desirable (P < 0.05) for overall eating quality than steaks from steers implanted two, three, four, or five times. Use of implants increased (P < 0.05) average daily gain by 11.8 to 20.5% from weaning to harvest compared with nonimplanted controls. Implant strategies increased (P < 0.05) hot carcass weight of steers by 8.9 to 13.8% compared with the control group. Use of implants also increased (P < 0.05) longissimus muscle area and decreased (P < 0.05) estimated percentages of kidney/pelvic/heart fat, but did not affect (P > 0.05) dressing percentage or adjusted fat thickness. Our findings suggest that beef quality, palatability, and production characteristics are influenced by lifetime implant protocols.     

Effects of a growth hormone-releasing factor analogue and an estradiol-trenbolone acetate implant on somatotropin, insulin-like growth factor I, and metabolite profiles in growing Hereford steers.

Hereford steers (290 +/- 6 kg of BW) were implanted (n = 4) with 140 mg of trenbolone acetate (TBA) and 28 mg of estradiol-17 beta (E2 beta) or nonimplanted (controls, n = 4). In Trial 1, effects of a single i.v. injection of 0, 20, 40, or 80 micrograms of a growth hormone-releasing factor (1-29 NH2) analogue (GRFa) on release of endogenous somatotropin (ST) were evaluated in a double 4 x 4 Latin square design. Plasma samples (n = 21) were obtained from -20 to 240 min after GRFa injection. Area under the ST response curve (AUC) increased (P = .009) in a dose-dependent manner (.2, 2.6, 3.6, 4.3 mg.min-1.mL-1, respectively). Mean ST concentration was not affected (P = .238) by implant but AUC was greater (P = .009) in implanted than in control steers. There was no interaction (P = .460) between dose of GRFa and presence of implant. In Trial 2, 80 micrograms of GRFa was administered at 12-h intervals to the same eight steers. Response of ST (AUC) to the first and last (13th) i.v. injection of GRFa was similar and not affected by implant. Before GRFa administration, plasma insulin-like growth factor I (IGF-I) concentrations were greater (P = .039) in implanted than in control steers (272 vs 164 ng/mL). Administration of GRFa increased plasma IGF-I (P = .0001), decreased plasma urea N (PUN) (P = .0001), and did not alter plasma glucose (P = .447) in both control and implanted steers. Data indicate that effects of GRFa and TBA/E2 beta on plasma IGF-I and PUN concentrations were additive in this study.     

Effects of L-carnitine on fetal growth and the IGF system in pigs

The effects of L-carnitine on porcine fetal growth traits and the IGF system were determined. Fourth-parity sows were fed a gestation diet with either a 50-g top dress containing 0 (control, n = 6) or 100 mg of L-carnitine (n = 6). At midgestation, fetuses were removed for growth measurements, and porcine embryonic myoblasts (PEM) were isolated from semitendinosus. Real-time quantitative PCR was used to measure growth factor messenger RNA (mRNA) levels in the uterus, placenta, muscle, hepatic tissue, and cultured PEM. A treatment x day interaction (P = 0.02) was observed for maternal circulating total carnitine. Sows fed L-carnitine had a greater (P = 0.01) concentration of total carnitine at d 57 than control sows. Circulating IGF-I was not affected (P = 0.55) by treatment. Supplementing sows with L-carnitine resulted in larger (P = 0.02) litters (15.5 vs. 10.8 fetuses) without affecting litter weight (P = 0.07; 1,449.6 vs. 989.4 g) or individual fetal weight (P = 0.88) compared with controls. No treatment effect was found for muscle IGF-I (P = 0.36), IGF-II (P = 0.51), IGFBP-3 (P = 0.70), or IGFBP-5 (P = 0.51) mRNA abundance. The abundance of IGF-I (P = 0.72), IGF-II (P = 0.34), and IGFBP-3 (P = 0.99) in hepatic tissue was not influenced by treatment. Uterine IGF-I (P = 0.46), IGF-II (P = 0.40), IGFBP-3 (P = 0.29), and IGFBP-5 (P = 0.35) mRNA abundance did not differ between treatments. Placental IGF-I (P = 0.30), IGF-II (P = 0.18), IGFBP-3 (P = 0.94), and IGFBP-5 (P = 0.42) mRNA abundance did not differ between treatments. There was an effect of side of the uterus for IGF-I (P = 0.04) and IGF-II (P = 0.007) mRNA abundance; IGF-I mRNA abundance was greater in the left uterine horn than in the right uterine horn (0.14 and 0.07 relative units, respectively). Placental IGF-II mRNA abundance was greater (P = 0.007) in the left than in the right uterine horn (483.5 and 219.59, respectively). The abundance of IGFBP-3 was not affected by uterine horns in either uterine (P = 0.66) or placental (P = 0.13) tissue. There was no treatment difference for IGF-I (P = 0.31) or IGFBP-5 (P = 0.13) in PEM. The PEM isolated from sows fed L-carnitine had decreased IGF-II (P = 0.02), IGFBP-3 (P = 0.03), and myogenin (P = 0.04; 61, 59, and 67%, respectively) mRNA abundance compared with controls. These data suggest that L-carnitine supplemented to gestating sows altered the IGF system and may affect fetal growth and development.     

Analysis of DNA Methylation and mRNA Expression Level of MSTN Gene in Xinjiang Bashiby Sheep

In order to investigate DNA methylation and expression levels of myostatin (MSTN) gene in mscule and fat, 5 months of age of Bashiby sheep were selected, the promoter region and exon 1 methylation levels of MSTN gene was analyzed using bisulfite sequencing PCR (BSP). Real-time PCR was used to detect the expression level of MSTN gene in biceps femoris, femoral triceps, semitendinosus, semimembranosus, longissimus dorsi muscle and tail fat of Bashiby sheep. The results showed that the methylation probability of muscle was higher than fat, the methylation probability of biceps femoris, femoral triceps, semitendinosus, semimembranosus, longissimus dorsi muscle and tail fat were 74.2%, 74.2%, 83.2%, 83.7%, 82.1% and 25.3%, respectively. The expression levels of MSTN gene in biceps femoris, femoral triceps, semitendinosus, semimembranosus, longissimus dorsi muscle were significantly lower than tail fat in Bashiby sheep (P < 0.05), but there were no significant difference among biceps femoris, femoral triceps, semitendinosus, semimembranosus and longissimus dorsi muscle (P > 0.05).The DNA methylation was negatively correlated with the expression levels in muscle and fat of Bashiby sheep (r=-0.886, P < 0.05).     

Pregnancy-associated plasma protein-A and insulin-like growth factor binding protein mRNAs in granulosa cells of dominant and subordinate follicles of preovulatory cattle

To determine if (1) levels of pregnancy-associated plasma protein-A (PAPP-A) mRNA and insulin-like growth factor binding protein (IGFBP) (-2, -3, -4 and -5) mRNAs differ between the dominant and subordinate follicles during the follicular phase of an estrous cycle, and (2) these differences are associated with differences in follicular fluid (FFL) concentrations of steroids (estradiol, androstenedione, and progesterone), total and free IGF-I, or IGFBPs, estrous cycles of non-lactating Holstein dairy cows (n = 16) were synchronized with two injections of prostaglandin (PGF2 alpha) 11 days apart. Granulosa cells and FFL were collected either 24 h or 48 h after the second injection of PGF2 alpha. FFL from dominant follicles had lower concentrations of progesterone (P < 0.08) and higher concentrations of estradiol (P < 0.05), androstenedione (P < 0.0001), estradiol:progesterone ratio (P < 0.0001), free IGF-I (P < 0.0001), and calculated percentage free IGF-I (P < 0.01) than large subordinate follicles. Levels of IGFBP-2, -4, and -5 in FFL were 3.0- (P < 0.05), 2.4- (P < 0.06), and 3.4-fold (P < 0.05) greater, respectively, in subordinate than in dominant follicles. IGFBP-3, IGFBP-4 and PAPP-A mRNA expression and IGF-II concentration did not differ (P > 0.10) between dominant or subordinate follicles. Levels of IGFBP-2 and -5 mRNA were severalfold greater (P < 0.05) in subordinate than dominant follicles. IGFBP-5 mRNA in granulosa cells decreased (P < 0.05) 62% to 92%, between 24h and 48 h post-PGF2 alpha. We conclude that decreased levels of IGFBP-2 and -5 mRNA in granulosa cells may contribute to the decrease in FFL IGFBP-2 and -5 protein levels of preovulatory dominant follicles, and that changes in granulosa cell IGFBP-3 and -4 mRNA and PAPP-A mRNA levels do not occur during final preovulatory follicular development in cattle.     

Effects of implant regimens (trenbolone acetate-estradiol administered alone or in combination with zeranol) and vitamin D3 on fresh beef color and quality

In the first oftwo experiments, 123 calf-fed steers were used over a 2-yr period to evaluate the effects of trenbolone acetate (TBA)-based implants administered alone or in combination with zeranol implants on fresh beef muscle quality, color, and physiological maturity of the carcass. Implant treatments decreased (P < 0.05) a* values (d 0 and d 3 of retail display) and b* values (d 0, d 1, and d 3 of retail display) after 14 d of aging. Carcasses from cattle initially implanted with Revalor-S and reimplanted with Revalor-S on d 60 of the finishing period showed increased lean and bone maturity scores and ash content of the 9th to 11th thoracic buttons and Warner-Bratzler shear force values (WBS) compared to those initially implanted with Ralgro and subsequently reimplanted with Revalor-S or control cattle. In addition, implants decreased (P < 0.05) marbling, percentage of the carcasses grading Choice, and kidney, pelvic, and heart fat (KPH). Implant treatments increased (P < 0.05) ADG, hot carcass weights, and longissimus muscle (LM) area. In the second experiment over a 2-yr period, 166 steers fed as yearlings were allotted to one of two implant treatments and one of two vitamin D3 preharvest supplementation treatments. Implanted steers had heavier (P < 0.05) final body weights and higher (P < 0.05) ADG, less (P < 0.05) KPH fat, and larger (P < 0.05) LM. Also, implanted steers had more (P < 0.05) advanced bone maturity scores, higher (P < 0.05) ash content of the 9th to 11th thoracic buttons, and higher (P < 0.05) WBS values on 5-d postmortem loin steaks. Vitamin D3 feeding decreased (P < 0.05) final live weight, ADG (P < 0.05), and LM (P < 0.05), but did not significantly improve WBS values. In Experiment 2, neither implant treatment nor vitamin D3 supplementation had significant effects on L*, a*, or b* values of muscles in steaks before or during simulated retail display.     

Growth factor messenger ribonucleic acid expression during differentiation of porcine embryonic myogenic cells

The growth factors, IGF-I and II, their binding proteins, IGFBP, and members of the transforming growth factor (TGF) superfamily (myostatin and TGFbeta1) are known to regulate proliferation and differentiation of myogenic cells. We hypothesized that changes in the relative expression of members of the IGF and TGFbeta systems play a significant role in regulating myogenesis in porcine embryonic myogenic cell (PEMC) cultures. Therefore, determining the expression patterns of these factors during PEMC myogenesis is important. Consequently, we used real-time PCR to explore the pattern of IGF-I; IGF-II; IGFBP-2, -3, and -5; IGF-type-I receptor; myogenin; myostatin; and TGFbeta1 mRNA expression during PEMC myogenesis. The progression of differentiation was assessed using creatine kinase activity and myogenin mRNA expression. As anticipated, creatine kinase activity was low in PEMC cultures at 48 h and increased 20-fold (P < 0.0001) between 48 h and its peak at 144 h. Similarly, myogenin mRNA was low at 48 h and increased approximately 5-fold (P < 0.0001) as differentiation progressed, peaking at 120 h and decreasing at 144 h. The patterns of IGF-I and IGFBP-2 mRNA expression were similar and were relatively lower in 48-h PEMC cultures, increasing approximately 5-fold (P < 0.0001) to their greatest levels at 120 h. In contrast, IGF-II and IGFBP-5 mRNA levels were relatively high at 48 h, peaking at 72 h, and steadily decreasing by 60 and 80%, respectively (P < 0.001), at 144 h. The level of IGF-type-I receptor mRNA was relatively high until 96 h of culture, after which it decreased 40% (P < 0.01), reaching a low at 144 h. Levels of IGFBP-3 mRNA were relatively high at 48 h, dropped approximately 40% to their lowest level at 72 h (P < 0.001), and then increased approximately 60% (P < 0.001) to their greatest levels at 144 h. Levels of TGFbeta1 mRNA decreased approximately 30% (P < 0.0001) between 48 and 96 h, then quickly rebounded to a peak at 120 h, and by 144 h had dropped to the levels seen at 72 h. Myostatin mRNA was at its greatest level at 48 h and declined rapidly between 72 and 96 h, finally decreasing by approximately 80% at 144 h (P < 0.0001). Our data demonstrate that these factors are differentially regulated during PEMC myogenesis and provide new information about their pattern of mRNA expression in cultured porcine muscle cells.     

新疆巴什拜羊肌肉生长抑制素基因DNA甲基化与mRNA表达量分析

为了探究肌肉生长抑制素（myostatin,MSTN）基因在肌肉和脂肪中的DNA甲基化模式及mRNA表达水平,试验以5月龄巴什拜羊羔羊为研究对象,采用亚硫酸氢盐测序法（bisulfite sequencing PCR,BSP）检测MSTN基因启动子区和第1外显子甲基化模式,并通过实时荧光定量PCR检测MSTN基因在巴什拜羊股二头肌、股三头肌、半腱肌、半膜肌、背最长肌和尾脂中的mRNA表达水平。结果显示,肌肉组织甲基化概率高于脂肪组织,其中,股二头肌、股三头肌、半腱肌、半膜肌、背最长肌和尾脂的甲基化概率分别为74.2%、74.2%、83.2%、83.7%、82.1%和25.3%,MSTN基因在股二头肌、股三头肌、半腱肌、半膜肌、背最长肌中的表达水平显著低于尾脂（P < 0.05）,股二头肌、股三头肌、半腱肌、半膜肌和背最长肌之间差异不显著（P > 0.05）,巴什拜羊肌肉和脂肪MSTN基因DNA甲基化水平与MSTN基因表达量呈显著负相关（r=－0.886,P < 0.05）。     

Effect of feedlot management system on response to ractopamine-HCl in yearling steers

Two experiments evaluated the effects of conventional and natural feedlot management systems (MS) on ractopamine-HCl (RAC) response in yearling steers. Feedlot performance, carcass characteristics, skeletal muscle gene expression, and circulating IGF-I concentrations were measured. The conventional system included a combined trenbolone acetate and estradiol implant, Revalor-S (IMP), as well as monensin-tylosin feed additives (IA). Treatments were arranged in a 2 x 2 factorial and included: 1) natural (NAT): no IMP-no IA, no RAC; 2) natural plus (NAT+): no IMP-no IA, RAC; 3) conventional (CON): IMP-IA, no RAC; and 4) conventional plus (CON+): IMP-IA, RAC. In Exp. 1, one hundred twenty crossbred steers (initial BW = 400 +/- 26 kg) were allotted randomly to treatment in a randomized complete block design (BW was blocking criteria); pen was the experimental unit. In Exp. 2, twenty-four individually fed crossbred steers (initial BW = 452 +/- 25 kg) were used in a randomized complete block design (BW was blocking criteria) and assigned to the same treatments as Exp. 1, with 6 steers/treatment. In Exp. 2, serum was harvested on d 0 and 31 and within the 28-d RAC feeding period, at d 0, 14, and 28. Longissimus biopsy samples were taken on d 0, 14, and 28 of the RAC feeding period for mRNA analysis of beta-adrenergic receptors and steady-state IGF-I mRNA. In Exp. 1, ADG, G:F, final BW, and HCW were greatest for CON+ (P < 0.01). During the final 37 d, RAC increased ADG (P = 0.05) and increased overall G:F (P = 0.02). Marbling score was reduced (P = 0.02), and yield grade was improved with RAC (P = 0.02), but RAC did not affect dressing percentage (P = 0.96) or HCW (P = 0.31). In Exp. 2, MS x RAC interactions were detected in ADG and G:F the last 28 d, overall ADG and overall G:F, final BW, and HCW (P < 0.01). Dressing percentage, yield grade, and marbling score were not altered by MS or RAC (P > 0.10). Circulating IGF-I concentration was increased on d 31 by the conventional MS, and concentration was greater throughout the study than NAT steers (P < 0.01). Circulating IGF-I concentrations were not changed by RAC (P = 0.49). Abundance of beta(1)-AR mRNA tended to increase (P = 0.09) with RAC, but RAC did not affect beta(2)-AR, beta(3)-AR, or IGF-I mRNA (P > 0.40). Management system did not affect beta(1)-AR, beta(2)-AR, beta(3)-AR, or IGF-I mRNA (P > 0.18), yet a trend (P = 0.06) for MS x RAC for beta(2)-AR mRNA was detected. These results indicate that response to RAC is affected by feedlot management practices.