Browsing by Author "DeBoer, Michelle"

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    Advances in Horse Nutrition and Muscle Protein Synthesis: Grazing Warm Season Annual Forages and Evaluating Muscle Protein Synthesis in Response to Leucine
    (2016-08) DeBoer, Michelle
    Warm-season grasses have the potential to provide forage in Upper Midwest pastures. The objectives of this research were to determine yield, forage nutritive value and preference of annual warm-season grasses under horse grazing. Four adult horses (Equus caballus) grazed Japanese millet (Echinochloa esculenta (A. Braun) H. Sholz) Siberian millet (Setaria italica (L.) P. Beauv.), teff (Eragrostis tef (Zucc.) Trotter) sorghum sudan BMR (Sorghum bicolor (L.) Moench), sudangrass (Sorghum sudanense (Piper) Stapf.), and annual ryegrass (Lolium multiflorum L.) at mature and vegetative stages. Forages were seeded in two separate plots utilizing a randomized complete block design with three replicates per plot. Each plot was grazed at a distinct target maturity and each maturity was grazed a minimum of three times each grazing season over a two-year period. Sudangrass was consistently the highest yielding grass (P ≤ 0.0002), producing ≥ 5.5 MT ha-1 at the vegetative stage and ≥ 9.7 MT ha-1 at the mature stage while Japanese and Siberian millet exhibit the lowest yields at ≤ 5.5 MT ha-1 at the vegetative stage and ≤ 6.3 MT ha-1 at the mature stage. Annual ryegrass was the most preferred grass (P ≤ 0.0057) with ≥ 60% removal at the vegetative stage and ≥ 40% removal at the mature stage. Siberian millet was the least preferred grass with ≤ 40% removal at the vegetative stage and ≤ 5% removal at the mature stage. While warm-season grasses meet the nutritional requirements of many classes of horses, Ca:P under 1:1 was observed as well as high NO3-N levels. These conditions could lead to limited mineral availability and possible nitrate toxicity. While teff and sudangrass have potential to be grazed by horses, Ca:P and NO3-N levels should be determined before grazing. Limited research is available regarding the role of leucine in regulating equine skeletal muscle protein synthesis. The objective of this study was to evaluate the effect of leucine on protein synthesis in cultured equine satellite cells by evaluating translation initiation factors in a western blot, the incorporation of puromycin using a nonradioactive surface sensing of translation (SUnSET) method, and measuring the incorporation of [3H]Phenylalanine (3HPhe) in a protein synthesis assay. Satellite cells from equine semimembranosus muscle were grown in cell culture until they developed into myotubes. Treatments used in the western blot consisted of a no leucine control (CON), leucine (LEU), control plus rapamycin (CR), and leucine plus rapamycin (LR). LEU exhibited higher 4E-BP1 and rpS6 phosphorylation when compared to CON with no change observed in mTOR phosphorylation. No increase in phosphorylation was observed in CR or LR treatments. Puromycin incorporation was measured on treatments including a no puromycin control, a no leucine control (CON), and a leucine treatment (LEU). These results showed a 1.6-fold increase in puromycin incorporation between CON and LEU (P = 0.0004). Treatments used in the protein synthesis assay consisted of a leucine titration ranging from 0-nm to 408-nm. These results showed muscle protein synthesis rates increased as a result of leucine treatments with significant differences observed at 204-nM and 408-nM leucine compared to the untreated cells. These treatments exhibited a 1.6-fold increase in protein synthesis rates when compared to the untreated control (P ≤ 0.02). This study demonstrated the phosphorylation of translation initiation factors downstream of mTOR as well as increased 3HPhe and puromycin with leucine treatment. These results suggest leucine can trigger muscle protein synthesis in horses through activation of the mTOR pathway.
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    Forage quality and blood metabolites of horses grazing alfalfa, cool-season perennial grass, and teff
    (2018-07) DeBoer, Michelle
    The impact of forage species on blood metabolites concentrations of grazing horses (Equus caballus L.) is unknown. However, these differences can be crucial as plasma amino acid (AA) concentrations as well as the glucose and insulin response of grazing horses can be indicators of nutritional status or metabolic health. As a result, the objectives of these studies were to determine the impact of different forage species on plasma AA concentrations, protein synthesis, as well as the glucose and insulin response across seasons. Research was conducted in May (spring), July (summer), September (fall), and late October (late-fall) in St. Paul, MN in 2016. However, the data collected was divided into three different studies (1) July samples taken during the first 4 hours were used to evaluate the forage and plasma AA concentrations (2) samples collected in July and September were included in the glucose and insulin response analysis of the regular grazing season and (3) May and October samples were used to analyze the glucose and insulin response during the extended grazing season. Forage treatments included alfalfa (Medicago sativa L.), a mixed perennial cool-season grass (CSG) and teff (Eragrostis tef [Zucc.] Trotter), however, not all forage species were grazed every season. Alfalfa and CSG were grazed in May while CSG and teff were grazed during the October, with all three species grazed in July and September. During these times, forages were grazed by six horses (24 ± 2 yr) randomly assigned to a forage in either a Latin-square or cross-over design. Jugular catheters were inserted 1 h prior to the start of grazing and horses had access to pasture starting at 08:00 h for either 4 or 8 h depending on the season. Jugular venous blood samples were collected from each horse prior to being turned out (0 h) and then at 2 hour intervals following turnout. Plasma and serum samples were collected and analyzed for AA, glucose, and insulin. Corresponding forage samples were taken by hand harvest. Equine muscle satellite cell cultures were treated with sera from grazing horses to assess de novo protein synthesis. Seasons were analyzed separately and data were analyzed using the MIXED procedure in SAS with P ≤ 0.05. When evaluating forage, AA were generally lowest in teff and highest in CSG (P ≤ 0.05). Significant differences in threonine concentration in the plasma were observed; there was no effect on de novo protein synthesis of cultured equine myotubes treated with plasma obtained from the grazing horses (P ≥ 0.20). As a result, although there were significant differences in forage AA content only plasma threonine concentration was different at 4 h with no effect on protein synthesis of cultured equine satellite cells. When evaluating the glucose and insulin response during the regular grazing season, teff generally had lower (P ≤ 0.05) equine digestible energy (DE), crude protein (CP) and nonstructural carbohydrates (NSC) compared to the other forages. Differences in peak insulin were observed between horses grazing CSG and teff during the fall grazing (P ≤ 0.05). Additionally, when evaluating the extended grazing season, teff had lower NSC than CSG in the late-fall (P ≤ 0.05) with subsequently lower average glucose, average insulin, and peak insulin in horses grazing teff compared to CSG (P ≤ 0.05).These results suggest grazing teff could lower the glucose and insulin response of some horses, specifically in the fall and late-fall, and may provide an alternative forage for horses with metabolic concerns,