AMP-Activated Protein Kinase Abundance and Activation is increased with the β-Adrenergic agonist Zilpaterol Hydrochloride in Muscle from Feedlot Cattle

The objective of this research was to evaluate the effect of Zilpaterol Hydrochloride (ZH) on myogenic or adipogenic gene and protein expression in skeletal muscle. Two feeding trials and one cell culture experiments were conducted. Semimembranosus muscle tissue was collected from steers that had been fed a diet containing 8.3 mg of ZH/kg DM for the last 0 or 20 d of the finishing period with a 3 d withdrawal period. To test the mode of action an in vitromodel was used with, isolated bovine satellite cells isolated from muscle tissue. Real Time-QPCR (RTQPCR) was used to measure the relative mRNA abundance of Adenosine Monophosphate Protein Kinase α (AMPK), Myosin Heavy Chain (MHC) I, MHCIIA, MHCIIX, Insulin-like Growth Factor I (IGF-I), β-adrenergic receptor (βAR) 1 and 2, peroxisome proliferator-activated receptor gamma (PPARγ), and Stearoyl-CoA Desaturase (SCD). Western blotting was used to measure the relative protein abundance of AMPK and Phosphorylated-AMPK (pAMPK). No differences were detected in relative mRNA abundance of AMPK, MHCIIA, IGF-I, βAR1 and βAR2. However, MHCI, SCD, and PPARγ mRNA expression was decreased and MHCIIX mRNA increased from ZH fed cattle compared to non-ZH. For one experiment, AMPK protein expression increased, while in another experiment, AMPK phosphorylation increased with ZH fed animals. The increase in MHCIIX mRNA with ZH fed cattle indicated the start of a fiber type shift towards larger diameter fibers. This shift may have been due to increased expression and phosphorylation of AMPK. These data suggest that the shift increase in MHCIIX was likely due to the ZH administration.

Meat Quality of Nellore Young Bulls—Effects of Different Days on Feed and Zilpaterol Hydrochloride Supplementation

Ninety-six Nellore young bulls were fed (90 or 117 day) diets containing ZH (8.33 mg/kg) for 0, 20, 30, or 40 days to evaluate the effects of days on feed (DOF) and length of zilpaterol hydrochloride (ZH) supplementation on meat quality. At the end of feeding period, animals were slaughtered, and samples of the Longissimus muscle were collected to evaluate the chemical composition, fatty acid profile, color stability, shear force, and sensory profile. DOF did not affect chemical composition, shear force, sensory tenderness, and most of fatty acids; however, animals fed for 90 d had lower redness (p < 0.01), sustained juiciness (p < 0.01), and more flavor (p = 0.03) than those fed for 117 d. The ZH supplementation decreased lipid content and redness (p < 0.01), initial and sustained tenderness (p < 0.01), initial and sustained juiciness (p < 0.01), but increased protein (p < 0.01) and shear force (p < 0.01) as compared to non-supplemented animals. The ZH supplementation increased total PUFA, c9,c12-18:2, and 20:4-n6, and decreased c9-20:1 (p < 0.05). Feeding ZH impairs meat quality attributes of Nellore young bulls, regardless of duration of supplementation, while DOF has a small effect on meat quality properties.

Effects of the Duration of Zilpaterol Hydrochloride Supplementation and Days on Feed on Performance, Carcass Traits and Saleable Meat Yield of Nellore Bulls

This study evaluated the effects of the duration of ZH supplementation and days on feed (DOF) on performance, carcass characteristics, and saleable meat yield of Nellore young bulls. The fixed effects included the duration (0, 20, 30, or 40 d before slaughter plus a 3 d ZH withdrawal period—8.33 mg of ZH/kg of DM) and DOF (90 and 117 d). Feed efficiency (G:F) linearly increased when the duration of ZH supplementation increased (p < 0.01). Nellore bulls fed ZH had greater HCW (p < 0.01), dressing percentage (p < 0.01) and Longissimus muscle area (LMA) (p < 0.01), but less 12th-rib fat (p = 0.04) than the control group. The hot carcass weight (HCW) (p < 0.01), and dressing percentage increased linearly (p < 0.01) with the increase of duration of ZH supplementation. The HCW, ossification, and 12th-rib fat increased with DOF (p < 0.01). The ZH supplemented group had most of the individual cuts of hindquarters and total saleable meat increased compared with the control. Zilpaterol hydrochloride was effective in improving hot carcass weight, hindquarter, and saleable meat yields of Nellore bulls when fed for at least 20 d before slaughter, independently of days on feed.

131 Zilpaterol Hydrochloride and Heat Stress Each Alter the Cattle Adipose Transcriptome and Predicted to Alter Molecular Pathways After 21 Days

Heat stress reduces livestock performance while supplementation of beta-adrenergic agonists (βAA) such as zilpaterol hydrochloride (ZH) improve production efficiency; both stimulate lipolysis. The objective of this study was to understand the effects (independent, interacting) of heat stress and ZH on the subcutaneous adipose transcriptome in fed steers. 24 Red Angus steers were assigned to thermoneutral (TN; THI=68) or heat stress (HS; THI=83) conditions and fed no supplement (NS) or ZH (8.33 mg/kg/day) for 21d in a 2x2 factorial. TN steers were pair-fed the HS daily average. Subcutaneous adipose samples were collected at day -3, 3, 10, and 21. RNA was isolated and sequenced using 3’ Tag-Seq reads to a depth of 3.6 million reads/sample. Transcripts were mapped to ARS-UCD1.2 and quantified. After quality control, differential expression (DE) analyses were performed in DESeq2 with a significance threshold (FRD) of 0.05. Pathway analysis was used to explore pathways affected by HS, ZH, and their interaction using DE loci (P &lt; 0.05). The acute phase response signaling pathway was predicted to be activated at 3d, but inhibited at 10d and 21d by the combination of HS and ZH. At multiple time points, inflammatory pathways including those for interferon and IL-8 were predicted to be activated by HS. Mitochondrial function pathways including oxidative phosphorylation, mitochondrial dysfunction, and TCA cycle II were altered by ZH. Adipose centric pathways for phospholipase-C and protein kinase-A signaling were altered by HS/ZH interaction while glycolysis was altered solely by HS. These data support the hypothesis that exposing cattle to HS conditions and ZH supplementation alters the subcutaneous adipose transcriptome, but not necessarily in an additive fashion. These data provide information regarding the supplementation of βAA in heat stress environments, especially if it mediates the effects of HS.