Application of Trimethylglycine Supplement in Broiler Chicken’s Diets with a High Dose of Dietary Guanidinoacetic Acid: Influence on Growth Performance and Physiological Variables

Document Type : Original Paper


Department of Animal Science, Shahrekord University, Shahrekord, 88186-34141, Iran


The present experiment studied the effect of a high dose of guanidinoacetic acid (GAA; 2.25 g/kg), with or without trimethylglycine (TMG) on growth performance and physiological variables of broiler chickens. A total of 300 day-old males Cobb 500 were randomly assigned to three dietary treatments with 5 replicates of 10 birds in each. Treatments were including a basal diet (control treatment), the basal diet supplemented with 2.25 g/kg GAA, and the GAA diet with 1 g/kg TMG. All birds received feed and water ad libitum during a 40-days rearing period. Results indicated that weight gain during days 31-40 of the rearing period was impaired by feeding the GAA diet. Feed conversion ratio was impaired in all feeding stages when the high dose of GAA was included in the diet. However, the inclusion of TMG in the GAA diet restored these responses to a comparable level to the control group. The GAA diet caused a significant increase in malondialdehyde concentration in serum compared to the control treatment. Moreover, the use of GAA decreased the heterophil to lymphocyte ratio than other treatments. A high dose of GAA caused higher serum levels of creatine and homocysteine; however, TMG supplementation re-established those responses. In addition, TMG supplement significantly up-regulated hepatic adenosyl homocysteinase and methionine adenosyltransferase II beta genes. In conclusion, feeding a high dose of GAA could impact broiler growth performance, but this effect could be ameliorated by dietary inclusion of TMG supplement, suggesting the negative effects of high doses of GAA were linked to the methyl donor deficiency. The practical implication is to include TMG in diets when a high dose of GAA is going to feed.


Ahmadipour B, Naeini SN, Sharifi M & Khajali F. 2018. Growth performance and right ventricular hypertrophy responses of broiler chickens to guanidinoacetic acid supplementation under hypobaric hypoxia. Journal of Poultry Science, 55: 60-64. DOI: 10.2141/jpsa.0170044
Behrooj N, Khajali F & Hassanpour H. 2012. Feeding reduced-protein diets to broilers subjected to hypobaric hypoxia is associated with the development of pulmonary hypertension syndrome. British Poultry Science, 53:658–664. DOI: 10.1080/00071668.2012.727082
Dorak M. 2006. Real Time PCR. Oxford, UK: Taylor & Francis.
Elango R. 2020. Methionine nutrition and metabolism: insights from animal studies to inform human nutrition. Journal of Nutrition, 150: 2518S-2523S. DOI: 10.1093/jn/nxaa155
Faraji M, Karimi Dehkordi S, Zamiani Moghadam AK, Ahmadipour B & Khajali F. 2019. Combined effects of guanidinoacetic acid, coenzyme Q10 and taurine on growth performance, gene expression and ascites mortality in broiler chickens. Journal of Animal Physiology and Animal Nutrition, 103: 162-169. DOI: 10.1111/jpn.13020
Jose MM, Fernando JC, Shelly CL & Mati AS. 2002. S-Adenosylmethionine: a control switch that regulates liver function. Federation of American Societies for Experimental Biology, 16: 15-26. DOI: DOI: 10.1096/fj.01-0401rev
Ibrahim D, El Sayed R, Abdelfattah-Hassan A & Morshedy AM. 2019. Creatine or guanidinoacetic acid? Which is more effective at enhancing growth, tissue creatine stores, quality of meat, and genes controlling growth/myogenesis in Mulard ducks. Journal of Applied Animal Research, 47: 159-166. DOI: 10.1080/09712119.2019.1590205
Kalia S, Bharti VK, Gogoi D, Giri A & Kumar B. 2017. Studies on the growth performance of different broiler strains at high altitude and evaluation of probiotic effect on their survivability. Scientific Reports 7: 1-8. DOI: 10.1038/srep46074
Khajali F, Lemme A & Rademacher-Heilshorn M. 2020. Guanidinoacetic acid as a feed supplement for poultry. World’s Poultry Science Journal, 76: 270-291. DOI: 10.1080/00439339.2020.1716651
Liu Y, Jia QZ, Han F, Inakuma T, Miyashita T, Sugiyama K, Sun LC, Xiang XS & Huang ZW. 2014. Suppression effects of trimethylglycine-enriched spinach on hyperhomocysteinemia induced by guanidinoacetic acid and choline deficiency in rats. Scientific World Journal, Article ID 904501. DOI: 10.1155/2014/904501
Lucas AM & Jamroz C. 1961. Atlas of Avian Hematology. Agriculture Monograph 25, United States Department of Agriculture.
McBreairty LE, Robinson JL, Furlong KR, Brunton JA & Bertolo R.F. 2015. Guanidinoacetate is more effective than creatine at enhancing tissue creatine stores while consequently limiting methionine availability in Yucatan miniature pigs. PLoS One 10: p.e0131563. DOI: 10.1371/journal.pone.0131563
Moghaddam AZ, Hamzekolaei MM, Khajali F & Hassanpour H. 2017. Role of selenium from different sources in prevention of pulmonary arterial hypertension syndrome in broiler chickens. Biological Trace Element Research, 180: 164-170. DOI: 10.1007/s12011-017-0993-3
Mori A, Kohno M, Masumizu T, Noda Y & Packer L. 1996. Guanidino compounds generate reactive oxygen species. Biochemistry and Molecular Biology International, 40: 135–143. DOI: 10.1080/15216549600201622
Nasiroleslami M, Torki M, Saki AA & Abdolmohammadi AR. 2018. Effects of dietary guanidinoacetic acid and trimethylglycine supplementation on performance, blood biochemical parameters and antioxidant status of broilers subjected to cold stress. Journal of Applied Animal Research, 46: 1016-1022. DOI: 10.1080/09712119.2018.1450751
Olthof MR & Verhoef P. 2005. Effects of trimethylglycine intake on plasma homocysteine concentrations and consequences for health. Current Drug Metabolism, 6: 15-22. DOI: 10.2174/1389200052997366
Ruijter J, Ramakers C, Hoogaars W, Karlen Y, Bakker O, Van den Hoff M & Moorman A. 2009. Amplification efficiency: Linking baseline and bias in the analysis of quantitative PCR data. Nucleic Acid Research, 37: 1–12. DOI: 10.1093/nar/gkp045
SAS Institute. 2008. SAS User’s guide: Statistics. Version 9.2. Cary, NC.
Sharma NK, Cadogan DJ, Chrystal PV, McGilchrist P, Wilkinson SJ, Inhuber V & Moss AF. 2021. Guanidinoacetic acid as a partial replacement to arginine with or without betaine in broilers offered moderately low crude protein diets. Poultry Science, 101692. DOI: 10.1016/j.psj.2021.101692 
Setoue M, Ohuchi S, Morita T & Sugiyama K. 2008. Hyperhomocysteinemia induced by guanidinoacetic acid is effectively suppressed by choline and trimethylglycine in rats. Bioscience, Biotechnology and Biochemistry, 72: 1696-1703. DOI: 10.1271/bbb.70791
Ueland PM, Holm PI & Hustad S. 2005. Trimethylglycine: a key modulator of one-carbon metabolism and homocysteine status. Clinical Chemistry and Laboratory Medicine, 43: 1069-1075. DOI: 10.1515/CCLM.2005.187
Vranes M, Ostojic S, Tot A, Papovic S & Gadzuric S. 2017. Experimental and computational study of guanidinoacetic acid self-aggregation in aqueous solution. Food Chemistry, 237: 53–57. DOI: 10.1016/j.foodchem.2017.05.088