Year of Publication

2016

Degree Name

Master of Science (MS)

Document Type

Master's Thesis

College

Agriculture, Food and Environment

Department

Animal and Food Sciences

First Advisor

Dr. Kristine Urschel

Abstract

Sarcopenia is a condition that is most common in aged animals, and is characterized by the loss of skeletal muscle mass and integrity, and can lead to physical disability and poor quality of life. Since skeletal muscle protein synthesis can be limited by the availability of amino acids, supplementation of limiting amino acids to ameliorate the progression of sarcopenia has become a topic of interest in companion animal research. Although there is some data to support the idea that amino acid supplementation improves maintenance of muscle mass in aged horses, the cellular mechanisms behind that improvement have yet to be elucidated. Therefore, the objective of this study was to examine the effect of amino acid supplementation in aged horses on markers of whole body and muscle protein metabolism. In a cross-over design, six old horses were studied while receiving each of three treatments in a replicated Latin square design. For all three treatments, horses received 1.8% BW/d of timothy hay cubes and 0.5% BW/d of experimental concentrate. The three treatments included a control (CON) treatment concentrate that was designed to meet all requirements of mature horses when fed in combination with the timothy hay cubes, and two supplemented concentrates, LYS/THR with additional lysine and threonine (40 mg/kg BW/d and 31 mg/kg BW/d, respectively), and LYS/THR/MET with additional lysine, threonine, and methionine (40 mg/kg BW/d, 31 mg/kg BW/d and 11mg/kg BW/d respectively). In each 15 d period, following a 9-day adaptation, horses were fitted with a collection harness, and total urine and feces were collected for 72 hours for assessment of nitrogen balance and creatinine output. Blood samples were taken directly before feeding and 30, 60, 90, 120, 150, 180, 210, and 240 minutes post-feeding for analysis of plasma urea nitrogen (PUN), glucose, insulin, and plasma amino acid concentrations. Muscle biopsy samples were taken for analysis of proteins in the mTOR pathway. Additionally, horses underwent stable isotope infusion procedures, and comparisons of phenylalanine kinetics were used to determine whole-body rates of protein synthesis and degradation. There was no significant effect of treatment on creatinine output (P=0.58), relative abundance of proteins in the mTOR pathway (P>0.05), nitrogen retention (P=0.70), or phenylalanine kinetics (P>0.05). PUN concentrations were significantly (P=0.0058) higher for LYS/THR and LYS/THR/MET than for CON. Atrogin-1 activation was significantly higher for the pre-feeding CON sample compared to the post-feeding CON sample. Lack of significant difference in creatinine output suggests that there were not significant differences in muscle mass between treatments. Lack of significant differences in mTOR protein activation suggests that amino acid supplementation did not result in improvements in protein synthesis. Lack of significant differences in nitrogen retention and phenylalanine kinetics suggests that whole-body protein metabolism was not improved. Additionally, higher PUN concentrations in the supplemented diets suggests that the supplemented amino acids being provided were catabolized. However, increased activation of Atrogin-1 in the pre-feeding CON samples, but not the pre-feeding samples of supplemented treatments, suggests amino acid supplementation may have reduced protein degradation in the post-absorptive state. Data from the present study suggests that amino acid availability may not have been limiting protein synthesis in the sedentary aged horses in the present study.

Digital Object Identifier (DOI)

http://dx.doi.org/10.13023/ETD.2016.142

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