AS  Vol.6 No.3 , March 2015
Effects of Dietary Soy Protein Concentrate on Growth, Digestive Enzymes Activities and Target of Rapamycin Signaling Pathway Regulation in Juvenile Soft-Shelled Turtle, Pelodiscus sinensis
Soft-shelled turtle, Pelodiscus sinensis is important aquatic species in China, and searching for alternatives protein resources to fish meal (FM)-based feeds in feed has become urgent and important for its sustainability development. The present study was conducted to assess the effects of dietary soy protein concentrate (SPC) on growth, digestive enzymes and target of rapamycin (TOR) signaling pathway of juvenile P. sinensis (4.56 ± 0.09 g). SPC was applied to replace FM protein at 0%, 15%, 30% and 60% (designated as T0, T15, T30 and T60, respectively), and each diet was fed to triplicate groups. The results showed that there was no significant difference in growth performance and feed utilization except of the turtles fed with T60 diet, of which showed poorer daily weight gain and feed conversion rate. The pepsin/trypsin and Na+-K+ ATP-ase activities decreased dramatically when SPC level increased, and lipase activities in liver and intestinal tract also showed decline tendency. However, amylase activities were unaffected. No significant differences were observed in TOR, S6K1 and 4E-BP1 genes mRNA expression level of TOR signaling pathway among the treatments. However, the relative phosphorylated level of these proteins decreased significantly when SPC level increased. The present study indicated that high SPC substitution level would suppress digestive enzymes and TOR signaling pathway proteins phosphorylated level and eventually result in growth reduction of P. sinensis.

Cite this paper
Zhou, F. , Wang, Y. , Tang, L. , Huang, Y. , Ding, X. and He, Z. (2015) Effects of Dietary Soy Protein Concentrate on Growth, Digestive Enzymes Activities and Target of Rapamycin Signaling Pathway Regulation in Juvenile Soft-Shelled Turtle, Pelodiscus sinensis. Agricultural Sciences, 6, 335-345. doi: 10.4236/as.2015.63034.
[1]   Zhou, F., Ding, X.Y., He, F., Xu, Y.B., Xue, H.L., Zhang, J.R. and Ng, W.K. (2013) The Dietary Protein Requirement of a New Japanese Strain of Juvenile Chinese Soft Shell Turtle, Pelodiscus sinensis. Aquaculture, 412-413, 74-80.

[2]   Jia, Y.Y., Yang, Z.C., Hao, Y.J. and Gao, Y.J. (2005) Efficacy and Tolerance of Lutein as Colourant in Diet of Juvenile Soft Shelled Turtle Pelodiscus sinensis. Aquaculture Research, 36, 61-68.

[3]   Hardy, R.W. (2010) Utilization of Plant Proteins in Fish Diets: Effects of Global Demand and Supplies of Fishmeal. Aquaculture Research, 41, 770-776.

[4]   Lin, S.M., Mai, K.S., Tan, B.P. and Liu, W.S. (2010) Effects of Four Vegetable Protein Supplementation on Growth, Digestive Enzyme Activities, and Liver Functions of Juvenile Tilapia, Oreochromis niloticus × Oreochromis aureus. Journal of the World Aquaculture Society, 41, 583-593.

[5]   Wullschleger, S., Loewith, R. and Hall, M.N. (2006) TOR Signaling in Growth and Metabolism. Cell, 124, 471-484.

[6]   Kim, J. and Guan, K.L. (2011) Amino Acid Signaling in TOR Activation. Annual Review of Biochemistry, 80, 1001-1032.

[7]   Hay, N. and Sonenberg, N. (2004) Upstream and Downstream of mTOR. Genes & Development, 18, 1926-1945.

[8]   Tee, A.R. and Blenis, J. (2005) mTOR, Translational Control and Human Disease. Seminars in Cell and Developmental Biology, 16, 29-37.

[9]   Lansard, M., Panserat, S., Seiliez, I., Polakof, S., Plagnes-Juan, E., Geurden, I., Medale, F., Kaushik, S., Corraze, G. and Skiba-Cassy, S. (2009) Hepatic Protein Kinase B (Akt)-Target of Rapamycin (TOR)-Signalling Pathways and Intermediary Metabolism in Rainbow Trout (Oncorhynchus mykiss) Are Not Significantly Affected by Feeding Plant- Based Diets. British Journal of Nutrition, 102, 1564-1573.

[10]   Lansard, M., Panserat, S., Plagnes-Juan, E., Seiliez, I. and Skiba-Cassy, S. (2010) Integration of Insulin and Amino Acid Signals That Regulate Hepatic Metabolism-Related Gene Expression in Rainbow Trout: Role of TOR. Amino Acids, 39, 801-810.

[11]   Seiliez, I., Panserat, S., Lansard, M., Polakof, S., Surget, A., Dias, K., Larquier, M., Kaushik, S. and Skiba-Cassy, S. (2011) Dietary Carbohydrate-to-Protein Ratio Affects TOR Signaling and Metabolism Related Gene Expression in the Liver and Muscle of Rainbow Trout after a Single Meal. American Journal of Physiology-Regulatory, Integrative and Comparative Physiology, 300, 733-743.

[12]   Zhao, J., Liu, Y., Jiang, J., Wu, P., Chen, G.F., Jiang, W.D., Li, S.H., Tang, L., Kuang, S.Y., Feng, L. and Zhou, X.Q. (2012) Effects of Dietary Isoleucine on Growth, the Digestion and Absorption Capacity and Gene Expression in Hepatopancreas and Intestine of Juvenile Jian Carp (Cyprinus carpio var. Jian). Aquaculture, 368-369, 117-128.

[13]   Feng, L., Peng, Y., Wu, P., Hu, K., Jiang, W.D., Liu, Y., Jiang, J., Li, S.H. and Zhou, X.Q. (2013) Threonine Affects Intestinal Function, Protein Synthesis and Gene Expression of TOR in Jian Carp (Cyprinus carpio var. Jian). PLoS ONE, 8, e69974.

[14]   Tang, L., Feng, L., Sun, C.Y., Chen, G.F., Jiang, W.D., Hu, K., Liu, Y., Jiang, J., Li, S.H., Kuang, S.Y. and Zhou, X.Q. (2013) Effect of Tryptophan on Growth, Intestinal Enzyme Activities and TOR Gene Expression in Juvenile Jian Carp (Cyprinus carpio var. Jian): Studies in Vivo and in Vitro. Aquaculture, 412-413, 23-33.

[15]   Luo, Y.W., Ai, Q.H., Mai, K.S., Zhang, W.B., Xu, W., Zhang, Y.J. and Liufu, Z.G. (2013) Effects of Dietary Corn Gluten Meal on Growth Performance and Protein Metabolism in Relation to IGF-I and TOR Gene Expression of Juvenile Cobia (Rachycentron canadum). Journal of Ocean University of China, 12, 418-426.

[16]   Sun, S.J., Liu, M., Peng, J.S., Wang, B.J., Jiang, K.Y. and Wang, L. (2010) Molecular Cloning of the TOR Gene from Fenneropenaeus chinensis and Its Expression in Response to Arginine or Leucine. Marine Sciences, 34, 71-80. (In Chinese)

[17]   Wang, Z., Pascual-Anaya, J., Zadissa, A., Li, W.Q., Niimura, Y., Huang, Z.Y., Li, C.Y., White, S., Xiong, Z.Q., Fang, D.M., Wang, B., Ming, Y., Chen, Y., Zheng, Y., Kuraku, S., Pignatelli, M., Herrero, J., Beal, K., Nozawa, M., Li, Q.Y., Wang, J., Zhang, H.Y., Yu, L.L., Shigenobu, S.J., Wang, J.Y., Liu, J.N., Flicek, P., Searle, S., Wang, J., Kuratani, S., Yin, Y., Aken, B., Zhang, G.J. and Irie, N. (2013) The Draft Genomes of Soft-Shell Turtle and Green Sea Turtle Yield Insights into the Development and Evolution of the Turtle-Specific Body Plan. Nature Genetics Letter, 45, 701-706.

[18]   Zhou, F., Wang, Y.Q., Ding, X.Y., Ng, W.K., He, F. and Xue, H.L. (2014) Partial Replacement of Fish Meal by Soy Protein Concentrates in Diets for a New Japanese Strain of Juvenile Soft-Shelled Turtle, Pelodiscus sinensis. Aquaculture Research, Early View.

[19]   AOAC (1995) Official Methods of Analysis. 15th Edition, Association of Official Analytical Chemists, Washington DC, 1094 p.

[20]   Deng, J.M., Mai, K.S., Ai, Q.H., Zhang, W.B., Wang, X.J., Xu, W. and Liufu, Z.G. (2006) Effects of Replacing Fish Meal with Soy Protein Concentrate on Feed Intake and Growth of Juvenile Japanese Flounder, Paralichthys olivaceus. Aquaculture, 258, 503-513.

[21]   Kissil, G.W., Lupatsch, I., Higgs, D.A. and Hardy, R.W. (2000) Dietary Substitution of Soy and Rapeseed Protein Concentrates for Fish Meal, and Their Effects on Growth and Nutrient Utilization in Gilthead Seabream Sparus aurata L. Aquaculture Research, 31, 595-601.

[22]   Day, O.J. and González, H.G.P. (2000) Soybean Protein Concentrate as a Protein Source for Turbot Scophthalmus maximus L. Aquaculture Nutrition, 6, 221-228.

[23]   Ronnestad, I., Kamisaka, Y., Conceicao, L.E.C., Morais, S. and Tonheim, S.K. (2007) Digestive Physiology of Marine Fish Larvae: Hormonal Control and Processing Capacity for Proteins, Peptides and Amino Acids. Aquaculture, 268, 82-97.

[24]   Leenhouwers, J.I., Adjei-Boateng, D., Verreth, J.A.J. and Schrama, J.W. (2006) Digesta Viscosity, Nutrient Digestibility and Organ Weights in African Catfish (Clarias gariepinus) Fed Diets Supplemented with Different Levels of a Soluble Non-Starch Polysaccharide. Aquaculture Nutrition, 12, 111-116.

[25]   López-López, S., Nolasco, H., Villarreal-Colmenares, H. and Civera-Cerecedo, R. (2005) Digestive Enzyme Response to Supplemental Ingredients in Practical Diets for Juvenile Freshwater Crayfish Cherax quadricarinatus. Aquaculture Nutrition, 11, 79-85.

[26]   Tibaldi, E., Hakim, Y., Uni, Z., Tulli, F., Francesco, M., Luzzana, U. and Harpaz, S. (2006) Effects of the Partial Substitution of Dietary Fish Meal by Differently Processed Soybean Meals on Growth Performance, Nutrient Digestibility and Activity of Intestinal Brush Border Enzymes in the European Sea Bass (Dicentrarchus labrax). Aquaculture, 261, 182-193.

[27]   Deng, J.M., Mai, K.S., Ai, Q.H., Zhang, W.B., Tan, B.P., Xu, W. and Liufu, Z.G. (2010) Alternative Protein Sources in Diets for Japanese Flounder Paralichthys olivaceus (Temminck and Schlegel): II. Effects on Nutrient Digestibility and Digestive Enzyme Activity. Aquaculture Research, 41, 861-870.

[28]   Kumar, V., Makkar, H.P.S. and Becker, K. (2011) Detoxified Jatropha curcas Kernel Meal as a Dietary Protein Source: Growth Performance, Nutrient Utilization and Digestive Enzymes in Common Carp (Cyprinus carpio L.) Fingerlings. Aquaculture Nutrition, 17, 313-326.

[29]   Fontagné, S., Burtaire, L., Corraze, G. and Bergot, P. (2000) Effects of Dietary Medium-Chain Triacylglycerols (Tricaprylin and Tricaproin) and Phospholipid Supply on Survival, Growth and Lipid Metabolism in Common Carp (Cyprinus carpio L.) Larvae. Aquaculture, 190, 289-303.

[30]   Escaffre, A.M., Infante, J.L.Z., Cahu, C.L., Mambrini, M., Bergot, P. and Kushik, S.J. (1997) Nutritional Value of Soy Protein Concentrate for Larvae of Common Carp (Cyprinus carpio) Based on Growth Performance and Digestive Enzyme Activities. Aquaculture, 153, 63-80.

[31]   Gal-Garber, O., Mabjeesh, S.J., Sklan, D. and Uni, Z. (2003) Nutrient Transport in the Small Intestine: Na+,K+-ATPase Expression and Activity in the Small Intestine of the Chicken as Influenced by Dietary Sodium. Poultry Science, 82, 1127-1133.

[32]   Krogdahl, ?., Bakke-McKellep, A.M. and Baeverfjord, G. (2003) Effects of Graded Levels of Standard Soybean Meal on Intestinal Structure, Mucosal Enzyme Activities, and Pancreatic Response in Atlantic Salmon (Salmo salar L.). Aquaculture Nutrition, 9, 361-371.

[33]   Lin, S.M. and Luo, L. (2011) Effects of Different Levels of Soybean Meal Inclusion in Replacement for Fish Meal on Growth, Digestive Enzymes and Transaminase Activities in Practical Diets for Juvenile Tilapia, Oreochromis niloticus × O. aureus. Animal Feed Science and Technology, 168, 80-87.

[34]   Mommsen, T.P. (2001) Paradigms of Growth in Fish. Comparative Biochemistry and Physiology Part B, 129, 207-219.

[35]   Rhoads, J.M., Cori, B.A., Harrell, R., Niu, X.M., Gatlin, L., Phillips, O., Blikslager, A., Moeser, A., Wu, G. and Odle, J. (2007) Intestinal Ribosomal p70S6K Signaling Is Increased in Piglet Rotavirus Enteritis. American Journal of Physiology-Gastrointestinal and Liver Physiology, 292, 913-922.

[36]   Wang, X. and Proud, C.G. (2006) The mTOR Pathway in the Control of Protein Synthesis. Physiology, 21, 362-369.

[37]   Wacyk, J., Powell, M., Rodnick, K., Overturf, K., Hill, R.A. and Hardy, R. (2012) Dietary Protein Source Significantly Alters Growth Performance, Plasma Variables and Hepatic Gene Expression in Rainbow Trout (Oncorhynchus mykiss) Fed Amino Acid Balanced Diets. Aquaculture, 356-357, 223-234.

[38]   Mehler, A.H. (1986) Amino Acid Metabolism II: Metabolism of the Individual Amino Acids. In: Devlin, T.M., Ed., Textbook of Biochemistry: With Clinical Correlations, John Wiley & Sons Inc, New York, 462-464.

[39]   Seiliez, I., Gabillard, J.C., Garcia-Serrana, S., Gutiérrez, D., Kaushik, J., Panserat, S. and Tesseraud, S. (2008) An in Vivo and in Vitro Assessment of TOR Signaling Cascade in Rainbow Trout (Oncorhynchus mykiss). American Journal of Physiology. Regulatory, Integrative and Comparative Physiology, 295, 329-335.

[40]   Payne, E.M., Virgilio, M., Narla, A., Sun, H., Levine, M., Paw, B.H., Berliner, N., Look, A.T., Ebert, B.L. and Khanna-Gupta, A. (2012) L-Leucine Improves the Anemia and Developmental Defects Associated with Diamond-Blackfan Anemia and del(5q) MDS by Activating the mTOR Pathway. Blood, 120, 2214-2224.

[41]   Abreu, R.S., Penalva, L.O., Marcotte, E.M. and Vogel, C. (2009) Global Signatures of Protein and mRNA Expression Levels. Molecular BioSystems, 5, 1512-1526.

[42]   Price, D.J., Grove, J.R., Calvo, V., Avruch, J. and Bierer, B.E. (1992) Rapamycin-Induced Inhibition of the 70-Kilodalton S6 Protein Kinase. Science, 257, 973-977.