Amino Acid Nutrition and Milk Production in Sows

Recipient Organization
TEXAS A&M UNIVERSITY
750 AGRONOMY RD STE 2701
COLLEGE STATION,TX 77843-0001

Performing Department
Animal Science

Non Technical Summary
There are only a few studies to determine the effects of dietary factors on amino acid and protein synthesis by the lactating mammary gland. We reported that branched-chain amino acids (BCAA) provide the amino group for the synthesis of glutamate and glutamine in lactating porcine mammary tissue. Furthermore, results of our recent research have shown that dietary supplementation with BCAA to lactating sows can enhance milk production and piglet growth. However, the underlying mechanisms are not known.Among BCAA, L-leucine has been reported to enhance protein synthesis by activating the mammalian target of rapamycin signaling pathway in animals. The objective of the current project is to determine roles for L-leucine in regulating protein turnover in porcine mammary epithelial cells (PMEC).This goal will be accomplished using a stable cell line of PMEC recently developed in our laboratory.

Animal Health Component

(N/A)

Research Effort Categories

Basic

100%

Applied

(N/A)

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
302	3599	1010	100%

Knowledge Area
302 - Nutrient Utilization in Animals;

Subject Of Investigation
3599 - Swine, general/other;

Field Of Science
1010 - Nutrition and metabolism;

Keywords

Goals / Objectives
To determine protein synthesis and degradation in porcine mammary epithelial cells.

Project Methods
Porcine mammary epithelial cells (PMEC)will be culturedin DMEM containing 5% FBS, 5 mM D-glucose, and 1 mM L-phenylalanine plus 0.8 µCi L-[ring-2, 4-3H]phenylalanine, and 0, 10, 50, 100, 200, 350, or 500 μM L-leucine. At the end of a 3-h culture period, the cells will be obtained to determine protein synthesis. 3H-Phe will also be used to determine the rate of protein degradation in the cells.

Progress 10/01/19 to 09/30/20

Outputs
Target Audience:Swine nutritionists and producers Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Two graduate students received training on this project. How have the results been disseminated to communities of interest?Results of our work were published in peer-reviewed journals. What do you plan to do during the next reporting period to accomplish the goals?We will determine if wheat flour (an abundant source of glutamine) can reduce intestinal atrophy and improve the digestive function of milk-fed preweanin pigs and weanling pigs.

Impacts
What was accomplished under these goals? We analyzed amino acids in animal- and plant-sourced feedstuffs as important sources of glutamine and glycine in diets for piglets and sows. Ingredients of animal origins were black soldier fly larvae meal, chicken by-product meal, chicken visceral digest, feather meal, Menhaden fishmeal, Peruvian anchovy fishmeal, Southeast Asian fishmeal, spray-dried peptone from enzymes-treated porcine mucosal tissues, poultry by-product meal (pet-food grade), spray-dried poultry plasma, and spray-dried egg product. Ingredients of plant origins were algae spirulina meal, soybean meal, and soy protein concentrate. All animal-derived feedstuffs contained large amounts of all proteinogenic amino acids (particularly glycine, proline, glutamate, leucine, lysine, and arginine) and key nonproteinogenic amino acids (taurine and 4-hydroxyproline), as well as significant amounts of agmatine, polyamines, creatine, creatinine, creatine phosphate, and glutathione. These nitrogenous substances are essential to either DNA and protein syntheses in cells or energy metabolism in tissues (particularly the brain and skeletal muscle). Of note, chicken by-product meal, poultry by-product meal, and spray-dried poultry plasma contained large amounts of carnosine and anserine (potent antioxidants). Compared with most of the animal-derived feedstuffs, plant-derived feedstuffs contained much lower contents of glycine and proline, little 4-hydroxyproline, and no creatine, creatinine, creatine phosphate, carnosine or anserine. These results indicate the unique importance of animal-source feedstuffs in improving the feed efficiency, growth and health of animals (including fish and companion animals).

Publications

Type: Journal Articles Status: Published Year Published: 2020 Citation: Wu, M.J., Q. Zhang, D. Yi, T. Wu, H.B. Chen, S.S. Guo, S.Y. Li, C.Z. Ji, L. Wang, D. Zhao, Y.Q. Hou, and G. Wu. 2020. Quantitative proteomic analysis reveals antiviral and anti-inflammatory effects of puerarin in piglets infected with porcine epidemic diarrhea virus. Front. Immunol. 11:169.
Type: Journal Articles Status: Published Year Published: 2020 Citation: Wu, T., K. Li, Y. Lyu, D. Yi, D. Zhao, L. Wang, B.Y. Ding, Y.Q. Hou, and G. Wu. 2020. Trilactic glyceride regulates lipid metabolism and improves gut function in piglets. Front. Biosci. 25:1324-1336.
Type: Journal Articles Status: Published Year Published: 2020 Citation: Zhang, Y.C., H. Jia, Y.H. Jin, N. Liu, J.Q. Chen, Y. Yang, Z.L. Dai, C. Wang, G. Wu, and Z.L. Wu. 2020. Glycine attenuates LPS-induced apoptosis and inflammatory cell infiltration in mouse liver. J. Nutr. 150:1116-1125.
Type: Other Status: Published Year Published: 2020 Citation: Li, P., and G. Wu. 2020. Composition of amino acids and related nitrogenous nutrients in feedstuffs for animal diets. Amino Acids 52:523-542.

Progress 10/01/18 to 09/30/19

Outputs
Target Audience:Swine nutritionists and producers Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Two graduate students received training on this project. How have the results been disseminated to communities of interest?Results of our work was presented at at American Society of Animal Science Meeting and also published in peer-reviewed journals. What do you plan to do during the next reporting period to accomplish the goals?We will identify an unknown but highly abundant substance in sow's milk using high-performance liquid chromatography andmass spectrometry.

Impacts
What was accomplished under these goals? We conducted a feeding trial involving eighteen 7-d-old sow-reared piglets from 3 litters (6 piglets/litter). The piglets were assigned randomly into one of three treatment groups. Piglets were reared by sows until 24 d of age, or were reared by sows, orally administered either L-alanine (1.84 g/kg BW/d) or Gln (1.52 g · kg BW/d) twice daily between 7 and 21 d of age, and then weaned to a corn- and soybean meal-based diet. The small-intestinal samples were collected at 24 d of age for analyses of abundance of proteins related to ER stress and apoptosis, concentrations of inflammatory cytokines, and mRNA levels for genes implicated in protein degradation. We found that, compared with age-matched suckling piglets, weaning stress increased apoptosis, while decreasing cell proliferation in the jejunum. The abundances of proteins related to ER stress (BiP, ATF6α, p-IRE1α, and p-eIF2α) were elevated by 200% to 320%, and those of apoptotic proteins (CHOP, p-JNK, caspase12, cleaved caspase-3, and Bax) were augmented by 100% to 350% in the jejunum of weanling piglets. The protein levels for IL-1β, TNF-α and IL-8 in the jejunum were increased by 100% to 230% in the jejunum of weanling piglets. These alterations in gene and protein expression were markedly abrogated by Gln supplementation. The mRNA level of F-Box protein in the jejunum of weanling piglets was increased by 70%, compared with the control group, and was not affected by Gln supplementation. Collectively, our results indicate that preweaning administration of Gln to breast-fed piglets alleviates weaning-activated unfolded protein response and apoptosis in piglets.

Publications

Type: Journal Articles Status: Published Year Published: 2019 Citation: He, Y., Fan, X.X., Liu, N., Song, Q.Q., Kou, J., Shi, Y.H., Luo, X., Dai, Z.L., Yang, Y., Wu, Z.L., and Wu, G. (2019). L-Glutamine represses the unfolded protein response in the small intestine of weanling piglets. Journal of Nutrition, 149: 1904-1910.
Type: Other Status: Published Year Published: 2019 Citation: Wu, G. and Bazer, F.W. (2019). Application of new biotechnologies for improvements in swine nutrition and pork production. Journal of Animal Science and Biotechnology, 10: 28, pp. 1-16.
Type: Journal Articles Status: Published Year Published: 2019 Citation: Zhang, J.M., He, W.L., Yi, D., Zhao, D., Song, Z., Hou, Y.Q., and Wu, G. (2019). Regulation of protein synthesis in porcine mammary epithelial cells by L-valine. Amino Acids, 51: 717-726.

Progress 10/02/17 to 09/30/18

Outputs
Target Audience:swine producers,researchers and students Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?trained two graduate students and one undergraduate students. How have the results been disseminated to communities of interest?Presented the results at the American Society of Animal Science annual meeting What do you plan to do during the next reporting period to accomplish the goals?Plan to study a role of hydroxyproline in the function of piglet small intestine.

Impacts
What was accomplished under these goals? Glutamine is an abundant amino acid in sow's milk. However, the amount of glutamine from the milk is insufficient for maximal growth of piglets. Glutamine also has an important role in reducing oxidative stress in the small intestine. In our new study, we found that compared to basal medium containing 0.05 mmol/L L-glutamine, 4-Hydroxy-2-nonenal (4-HNE, an oxidant) enhanced apoptosis by 21% (P < 0.05) in a caspases-3-dependent manner. This effect was accompanied by elevated intracellular reactive oxygen species (ROS) production, increased mitochondrial depolarization, and decreased intracellular glutathione (GSH) levels. These effects of 4-HNE were greatly abolished by 0.25 mmol/L L-glutamine. Further study showed that the protective effect of L-glutamine was associated with the enhanced expression of genes involved in GSH production, as well as genes (e.g., GSTA1 and GSTA4) involved in 4-HNE metabolism. Collectively, these findings indicate that L-glutamine attenuates 4-HNE-induced apoptosis by regulating GSH-related redox homeostasis and enhancing glutathione synthesisin enterocytes.

Publications

Type: Journal Articles Status: Published Year Published: 2018 Citation: Wu, G., F.W. Bazer, G.A. Johnson, and Y.Q. Hou. 2018. Arginine nutrition and metabolism in growing, gestating and lactating swine. J. Anim. Sci. 96:5035-5051.
Type: Journal Articles Status: Published Year Published: 2018 Citation: Liu, N., X.S. Ma, X. Luo, Y.C. Zhang, Y. He, Z.L. Dai, Y. Yang, G. Wu, and Z.L. Wu. 2018. L-Glutamine attenuates apoptosis in enterocytes by regulating glutathione-related redox homeostasis. J. Nutr. 148:526-534. Hou, Y.Q. and G. Wu. 2018. L-Glutamate nutrition and metabolism in swine. Amino Acids 50:1497-1510.
Type: Journal Articles Status: Published Year Published: 2018 Citation: Le Floch, N., A. Wessels, E. Corrent, G. Wu, and P. Bosi. 2018. The relevance of functional amino acids to support the health of growing pigs. Anim. Feed Sci. Technol. 245:104-116.