Methionine (Met) is an essential amino acid for animals and also the first limiting amino acid in a broiler diet. The dietary supplemental Met sources include the natural isoform L-methionine (L-Met), the synthetic form DL-methionine (DL-Met) and the synthetic Met precursor DL-2-hydroxy-4 (methylthio)-butanoic acid (DL-HMTBA). The objective of this dissertation was to determine the effect of different dietary Met source supplementation and Met deficiency on a series of physiological and biochemical aspects, including growth performance, global DNA methylation and methyltransferase activity, blood antioxidant profile (e.g., acute phase protein, leukocyte count), intestinal nutrient transporter gene expression, Met converting enzyme gene expression and activity, oxidative stress markers, and a potential pathway related to amino acid signaling. To achieve this goal, male Cobb-500 broilers were raised from day of hatch (d0) to d35 post-hatch and fed a diet deficient in methionine and cysteine (Met + Cys) (control) or the same diet supplemented with 0.22% DL-Met, 0.22% L-Met or 0.31% DL-HMTBA (to provide 0.22% DL-Met equivalent) to meet Met + Cys requirements. Tissues (liver, breast muscle, duodenum, jejunum and ileum) and blood samples were collected at various ages from d0 to d35 for analysis. Met supplementation significantly improved body weight gain and feed efficiency compared to the Met deficient group, but no differences were observed among DL-Met, L-Met and DL-HMTBA for growth performance parameters (P > 0.05). Met supplementation had no effect on red blood cell packed cell volume, white blood cell differential count, hepatic total DNA methylation, DNA methyltransferase and Met oxidase activity, and had limited effects on activation of p70S6K, a key amino acid signaling protein (P > 0.05). Although dietary Met sources did not change oxidative status of the treated chickens, both L-Met and DL-Met but not DL-HMTBA supplementation decreased the level of acute phase protein serum amyloid A compared to the control group (P > 0.05). The effect of Met supplementation on gene expression of nutrient transporters and Met converting enzymes were complex and dynamic. Most of the target genes demonstrated tissue- and development-dependent expression patterns, with few significant treatment effects observed. L-Met and DL-Met but not DL-HMTBA supplementation enhanced the neutral amino acid transporters ATB0,+ and B0AT gene expression in various small intestinal segments. All three Met sources increased monocarboxylic acid transporter (MCT1) gene expression in the jejunum. DL-HMTBA and L-Met fed chickens showed greater hepatic L-HMTBA oxidase (HAO1) gene expression. DL-Met increased glutamic-oxaloacetic transaminase 2 (GOT2) gene expression in the duodenum. An in vitro study with tissue explants, however, did not demonstrate a similar gene expression pattern as that in the in vivo study. Lastly, RNA sequencing results illustrated that Met deficiency could lead to many differentially expressed genes but different supplemental Met sources had no influence on hepatic gene expression profiles. In conclusion, as common dietary supplemental Met sources, L-Met, DL-Met and DL-HMTBA exhibited similarity in impacting intestinal amino acid/peptide/monocarboxylic acid transporter gene expression and Met converting enzyme activity. The regulatory roles of Met as an antioxidant and nutrient signaling in cell metabolism were not affected by different dietary supplemental Met sources. / Ph. D.
| Identifer | oai:union.ndltd.org:VTETD/oai:vtechworks.lib.vt.edu:10919/71816 |
| Date | 19 July 2016 |
| Creators | Zhang, Shuai |
| Contributors | Animal and Poultry Sciences, Gilbert, Elizabeth R., Wong, Eric A., Corl, Benjamin A., Persia, Michael E. |
| Publisher | Virginia Tech |
| Source Sets | Virginia Tech Theses and Dissertation |
| Detected Language | English |
| Type | Dissertation |
| Format | ETD, application/pdf, application/pdf |
| Rights | In Copyright, http://rightsstatements.org/vocab/InC/1.0/ |
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