An Assessment of the Effects of Guanidinoacetic Acid on the Performance and Immune Response of Laying Hens Fed Diets with Three Levels of Metabolizable Energy.

Different levels of metabolizable energy (ME) and the inclusion of guanidinoacetic acid (GAA) in the diet of 53-week-old Lohmann LSL-CLASSIC hens were used to evaluate its effect on reproductive parameters, egg quality, intestinal morphology, and the immune response. Six diets were used in a 3 × 2 factorial design, with three levels of ME (2850, 2800, and 2750 kcal/kg), and with (0.08%) or without the inclusion of GAA. The addition of GAA to diets with low levels of ME increased (p < 0.05) egg production and egg mass. Moreover, hens fed with 2800 kcal/g without GAA had the highest concentration (p < 0.05) of serum interleukin IL-2, while those fed diets with the same amount of ME but supplemented with 0.08% GAA had the lowest concentration. Finally, the inclusion of 0.08% GAA increased (p < 0.05) the concentration of vascular endothelial growth factor (VEGF), regardless of the ME level in the diet. This study highlights the potential role of GAA in decreasing the energy level of ME (50-100 kcal/g) in the feeding of hens and in the modulation of specific immune responses. Further research is recommended to fully understand the mechanisms of action of GAA on the mechanism target of rapamycin and its relationship with the immune response.

Association analysis of transcriptome and quasi-targeted metabolomics reveals the regulation mechanism underlying broiler muscle tissue development at different levels of dietary guanidinoacetic acid.

The development and characteristics of muscle fibers in broilers are critical determinants that influence their growth performance, as well as serve as essential prerequisites for the production of high-quality chicken meat. Guanidinoacetic acid (GAA) is a crucial endogenous substance in animal creatine synthesis, and its utilization as a feed additive has been demonstrated the capabilities to enhance animal performance, optimize muscle yield, and augment carcass quality. The objective of this study was to investigate the regulation and molecular mechanism underlying muscle development in broilers at different levels of GAA via multiple omics analysis. The 90 Cobb broilers, aged 1 day, were randomly allocated into three treatments consisting of five replicates of six chickens each. The control group was provided with a basal diet, while the Normal GAA and High GAA groups received a basal diet supplemented with 1.2 g/kg and 3.6 g/kg of GAA, respectively. After a feeding period of 42 days, the pectoralis muscles were collected for histomorphological observation, transcriptome and metabolomic analysis. The results demonstrated that the addition of 1.2 g/kg GAA in the diet led to an augmentation in muscle fiber diameter and up-regulation of IGF1, IHH, ASB2, and ANKRD2 gene expression. However, a high dose of 3.6 g/kg GAA in the diet potentially reversed the beneficial effects on chicken breast development by excessively activating the TGF-ß signaling pathway and reducing nucleotide metabolite content. These findings would provide a theoretical foundation for enhancing the performance and meat quality of broilers by incorporating GAA as a feed additive.

Analysis of microRNA Expression Profiles in Broiler Muscle Tissues by Feeding Different Levels of Guanidinoacetic Acid.

The aim of this study was to explore the molecular mechanisms through which different levels of GAA affect chicken muscle development by influencing miRNA expression, to lay a theoretical foundation for the identification of key functional small RNAs related to poultry muscle development, and to provide new insights into the regulatory mechanisms of GAA on muscle development and meat quality in broilers. It provides a new theoretical basis for using GAA as a feed additive to improve feed performance. Small RNA sequencing technology was utilized to obtain the expression profiles of miRNA in the broiler pectoral muscle fed with different levels of GAA (0 g/kg, 1.2 g/kg and 3.6 g/kg). An analysis of differentially expressed miRNAs revealed 90 such miRNAs in the three combination comparisons, with gga-miR-130b-5p exhibiting significant differences across all three combinations. Furthermore, three of the differentially expressed miRNAs were performed by RT-qPCR verification, yielding results consistent with those obtained from small RNA sequencing. Target gene prediction, as well as the GO and KEGG enrichment analysis of differentially expressed miRNAs, indicated their involvement in muscle cell differentiation and other processes, particularly those associated with the MAPK signaling pathway. This study has, thus, provided valuable insights and resources for the further exploration of the miRNA molecular mechanism underlying the influence of guanidine acetic acid on broiler muscle development. Combined with previous studies and small RNA sequencing, adding 1.2 g/kg GAA to the diet can better promote the muscle development of broilers.

Effects of guanidinoacetic acid supplementation on liver and breast muscle fat deposition, lipid levels, and lipid metabolism-related gene expression in ducks.

Exogenous supplementation of guanidinoacetic acid can mechanistically regulate the energy distribution in muscle cells. This study aimed to investigate the effects of guanidinoacetic acid supplementation on liver and breast muscle fat deposition, lipid levels, and lipid metabolism-related gene expression in ducks. We randomly divided 480 42 days-old female Jiaji ducks into four groups with six replicates and 20 ducks for each replicate. The control group was fed the basal diet, and the experimental groups were fed the basal diet with 400, 600, and 800 mg/kg (GA400, GA600, and GA800) guanidinoacetic acid, respectively. Compared with the control group, (1) the total cholesterol (p = 0.0262), triglycerides (p = 0.0357), malondialdehyde (p = 0.0452) contents were lower in GA400, GA600 and GA800 in the liver; (2) the total cholesterol (p = 0.0365), triglycerides (p = 0.0459), and malondialdehyde (p = 0.0326) contents in breast muscle were decreased in GA400, GA600 and GA800; (3) the high density lipoprotein (p = 0.0356) and apolipoprotein-A1 (p = 0.0125) contents were increased in GA600 in the liver; (4) the apolipoprotein-A1 contents (p = 0.0489) in breast muscle were higher in GA600 and GA800; (5) the lipoprotein lipase contents (p = 0.0325) in the liver were higher in GA600 and GA800; (6) the malate dehydrogenase contents (p = 0.0269) in breast muscle were lower in GA400, GA600, and GA800; (7) the insulin induced gene 1 (p = 0.0326), fatty acid transport protein 1 (p = 0.0412), and lipoprotein lipase (p = 0.0235) relative expression were higher in GA400, GA600, and GA800 in the liver; (8) the insulin induced gene 1 (p = 0.0269), fatty acid transport protein 1 (p = 0.0234), and lipoprotein lipase (p = 0.0425) relative expression were increased in GA400, GA600, and GA800 in breast muscle. In this study, the optimum dosage of 600 mg/kg guanidinoacetic acid improved the liver and breast muscle fat deposition, lipid levels, and lipid metabolism-related gene expression in ducks.

Effect of Guanidinoacetic Acid Supplementation on Growth Performance, Rumen Fermentation, Blood Indices, Nutrient Digestion, and Nitrogen Metabolism in Angus Steers.

Guanidinoacetic acid (GAA) functions as a precursor for creatine synthesis in the animal body, and maintaining ample creatine reserves is essential for fostering rapid growth. This study aimed to explore the impact of GAA supplementation on growth performance, rumen fermentation, blood indices, nutrient digestion, and nitrogen metabolism in Angus steers through two experiments: a feeding experiment (Experiment 1) and a digestive metabolism experiment (Experiment 2). In Experiment 1, thirty-six Angus steers (485.64 ± 39.41 kg of BW) at 16 months of age were randomly assigned to three groups: control (CON), a conventional dose of GAA (CGAA, 0.8 g/kg), and a high dose of GAA (HGAA, 1.6 g/kg), each with twelve steers. The adaptation period lasted 14 days, and the test period was 130 days. Weighing occurred before morning feeding on days 0, 65, and 130, with rumen fluid and blood collected before morning feeding on day 130. Experiment 2 involved fifteen 18-month-old Angus steers (575.60 ± 7.78 kg of BW) randomly assigned to the same three groups as in Experiment 1, with a 7-day adaptation period and a 3-day test period. Fecal and urine samples were collected from all steers during this period. Results showed a significantly higher average daily gain (ADG) in the CGAA and HGAA groups compared to the CON group (p = 0.043). Additionally, the feed conversion efficiency (FCE) was significantly higher in the CGAA and HGAA groups than in the CON group (p = 0.018). The concentrations of acetate and the acetate:propionate ratio were significantly lower in the CGAA and HGAA groups, while propionate concentration was significantly higher (p < 0.01). Serum concentration of urea (UREA), blood ammonia (BA), GAA, creatine, and catalase (CAT) in the CGAA and HGAA groups were significantly higher than in the CON group, whereas malondialdehyde (MDA) concentrations were significantly lower (p < 0.05). Digestibility of dry matter (DM) and crude protein (CP) and the nitrogen retention ratio were significantly higher in the CGAA and HGAA groups than in the CON group (p < 0.05). In conclusion, dietary addition of both 0.8 g/kg and 1.6 g/kg of GAA increased growth performance, regulated rumen fermentation and blood indices, and improved digestibility and nitrogen metabolism in Angus steers. However, higher doses of GAA did not demonstrate a linear stacking effect.

Inclusion of guanidinoacetic acid in a low metabolizable energy diet improves broilers growth performance by elevating energy utilization efficiency through modulation serum metabolite profile.

This study was aimed to explore the elevating energy utilization efficiency mechanism for the potentially ameliorative effect of guanidinoacetic acid (GAA) addition on growth performance of broilers fed a low metabolizable energy (LME) diet. A total of 576 d old broilers were randomly allocated to one of the six treatments: a basal diet (normal ME, positive control, PC), or an LME diet (50 kcal/kg reduction in ME, negative control, NC) supplemented with 0.02%, 0.04%, 0.06%, and 0.08% GAA from 1 to 42 d of age, respectively. The GAA fortification in LME diet linearly or quadratically dropped (P < 0.05) the feed conversion ratio (FCR) from 22 to 42 and 1 to 42 d of age, abdominal fat rate on day 42, serum alanine aminotransferase (ALT) on day 21, and serum creatinine (CREAN) on days 21 and 42, elevated (P < 0.05) breast muscle rate and leg muscle rate on day 42, serum creatine kinase (CK) on days 21 and 42, as well as alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) on day 21. The dietary optimal GAA levels were 0.03%-0.08% based on the best-fitted quadratic models (P < 0.03) of the above parameters. Thus, the PC, LME, and 0.04% GAA-LME groups were selected for further analysis. Serum essential amino acids (EAA) tryptophan, histidine and arginine, non-essential amino acids (NEEA) serine, glutamine and aspartic acid were significantly decreased (P < 0.05), compared to PC diet by LME or 0.04% GAA-LME diet. 0.04% GAA-LME group reversed (P < 0.05) the reduction of arginine, 3-methyhistidine, and 1-methylhistidine by LME diet. Besides, six birds at 28 d of age from LME and 0.04% GAA-LME groups were selected for energy utilization observation in calorimetry chambers. The results demonstrated that 0.04% GAA-LME group significantly improved (P < 0.05) the ME intake (MEI) and net energy (NE) compared to the LME diet. Overall, these findings suggest that 0.04% GAA is the ideal dose of broilers fed the LME diet, which can significantly improve the growth performance and carcass characteristics by modulation of creatine metabolism through elevating serum CK activity and arginine concentration.

Guanidinoacetic acid (GAA) has been found to elevate energy utilization efficiency in broilers; however, the underlying mechanisms remain unclear. We investigated the effects of GAA addition in low metabolizable energy (LME) diet on growth performance, carcass characteristics and serum biochemical indices of broilers, and found that GAA addition linearly or quadratically dropped the feed conversion ratio from 22 to 42 and 1 to 42 d of age, abdominal fat rate on day 42, serum alanine aminotransferase on day 21, and serum creatinine on days 21 and 42, elevated breast muscle and leg muscle rate on day 42, serum creatine kinase, alkaline phosphatase, as well as lactate dehydrogenase on days 21 or 22. The dietary optimal GAA levels were 0.03%-0.08% based on the best-fitted quadratic models of the above parameters. Thus, further analysis was conducted and found that 0.04% GAA reversed the reduction of arginine, 3-methyhistidine, and 1-methylhistidine and improved the ME intake and net energy compared to the LME diet. These findings suggested that 0.04% GAA is the ideal dose for enhancing the energy utilization of broilers fed the LME diet, GAA addition can significantly improve the growth performance by elevating energy utilization efficiency through modulation serum metabolite profile.

Effect of Dietary Guanidinoacetic Acid Levels on the Mitigation of Greenhouse Gas Production and the Rumen Fermentation Profile of Alfalfa-Based Diets.

The objective of this study was to evaluate the effect of different percentages of alfalfa (Medicago sativa L.) hay (AH) and doses of guanidinoacetic acid (GAA) in the diet on the mitigation of greenhouse gas production, the in vitro rumen fermentation profile and methane (CH4) conversion efficiency. AH percentages were defined for the diets of beef and dairy cattle, as well as under grazing conditions (10 (AH10), 25 (AH25) and 100% (AH100)), while the GAA doses were 0 (control), 0.0005, 0.0010, 0.0015, 0.0020, 0.0025 and 0.0030 g g-1 DM diet. With an increased dose of GAA, the total gas production (GP) and methane (CH4) increased (p = 0.0439) in the AH10 diet, while in AH25 diet, no effect was observed (p = 0.1311), and in AH100, GP and CH4 levels decreased (p = 0.0113). In addition, the increase in GAA decreased (p = 0.0042) the proportion of CH4 in the AH25 diet, with no influence (p = 0.1050) on CH4 in the AH10 and AH100 diet groups. Carbon monoxide production decreased (p = 0.0227) in the AH100 diet with most GAA doses, and the other diets did not show an effect (p = 0.0617) on carbon monoxide, while the production of hydrogen sulfide decreased (p = 0.0441) in the AH10 and AH100 diets with the addition of GAA, with no effect observed in association with the AH25 diet (p = 0.3162). The pH level increased (p < 0.0001) and dry matter degradation (DMD) decreased (p < 0.0001) when AH was increased from 10 to 25%, while 25 to 100% AH contents had the opposite effect. In addition, with an increased GAA dose, only the pH in the AH100 diet increased (p = 0.0142 and p = 0.0023) the DMD in the AH10 diet group. Similarly, GAA influenced (p = 0.0002) SCFA, ME and CH4 conversion efficiency but only in the AH10 diet group. In this diet group, it was observed that with an increased dose of GAA, SCFA and ME increased (p = 0.0002), while CH4 per unit of OM decreased (p = 0.0002) only with doses of 0.0010, 0.0015 and 0.0020 g, with no effect on CH4 per unit of SCFA and ME (p = 0.1790 and p = 0.1343). In conclusion, the positive effects of GAA depend on the percentage of AH, and diets with 25 and 100% AH showed very little improvement with the addition of GAA, while the diet with 10% AH presented the best results.

Effects of guanidinoacetic acid supplementation on growth performance, hypothalamus-pituitary-adrenal axis, and immunity of broilers challenged with chronic heat stress.

Heat stress can cause systemic immune dysregulation and threaten the health of broilers. Guanidinoacetic acid (GAA) has been shown to be effective against heat stress, but whether it is beneficial for immunity is unclear. Therefore, the effects of dietary GAA supplementation on the immunity of chronic heat-stressed broilers were evaluated. A total of 192 Arbor Acres male broilers (28-day old) were randomly allocated to 4 treatments: the normal control group (NC, 22°C, ad libitum feeding), the heat stress group (HS, 32°C, ad libitum feeding), the pair-fed group (PF, kept at 22°C and received food equivalent to that consumed by the HS group on the previous day), and the GAA group (HG, 32°C, ad libitum feeding; basal diet supplemented with 0.6 g/kg GAA). Samples were collected on d 7 and 14 after treatment. Results showed that broilers exposed to heat stress exhibited a decrease (P < 0.05) in ADG, ADFI, thymus and bursa of Fabricius indexes, and an increase (P < 0.05) in feed conversion ratio and panting frequency, compared to the NC group. Levels of corticotropin-releasing factor, corticosterone (CORT), heat shock protein 70 (HSP70), IL-6, and TNF-α were elevated (P < 0.05) while lysozyme and IgG concentration was decreased (P < 0.05) in the HS group compared with the NC group after 7 d of heat exposure. The concentrations of IgG and IL-2 were decreased (P < 0.05) and CORT was increased (P < 0.05) in the HS group compared with the NC group after 14 d of heat exposure. Noticeably, GAA supplementation decreased the levels of CORT (P < 0.05) and increased the IL-2, IgG, and IgM concentrations (P < 0.05) compared with the HS group. In conclusion, chronic heat stress increased CORT release, damaged immune organs, and impaired the immunity of broilers. Dietary supplementation of 0.6 g/kg GAA can reduce the CORT level and improve the immune function of broilers under heat stress conditions.

Effect of guanidinoacetic acid on the growth performance, myofiber, and adenine nucleotide of meat-type rabbits.

OBJECTIVE: This study aimed to investigate the effect of dietary guanidinoacetic acid (GAA) on the growth performance, slaughter traits, myofiber, and adenine nucleotide of meat-type rabbits. METHODS: Experimental treatments consisted of control (CON) and GAA addition at 0.04% (T1), 0.08% (T2), and 0.12% (T3) of diet. A total of 240 weaned rabbits (meat-type male Chinese black rabbits) were randomly distributed into four groups with six replicates of ten rabbits each. RESULTS: Results showed that the three doses of GAA increased (p<0.05) final body weight, carcass weight, the density and area of quadriceps femoris fiber; and T3 showed significant effects (p<0.05) on weight gain, feed/gain, and dressing percentage, and the traits of longissimus fiber, compared to CON. Dietary GAA increased (p<0.05) the meat color a* and b* in longissimus and quadriceps; and T3 showed the lowest (p<0.05) shear force of longissimus. Furthermore, GAA increased (p<0.05) the contents of adenosine triphosphate and total adenine nucleotide in longissimus and quadriceps. In longissimus adenosine triphosphate, total adenine nucleotide, and adenylate energy charges, T3 treatment was most effective (p<0.05); while T2 and T3 treatment was more effective (p<0.05) than T1 in quadriceps. Additionally, linear or quadratic responses (p<0.05) to the increased doses of GAA were found on body weight gain, meat color, total adenine nucleotide, and adenylate energy charges. CONCLUSION: It is concluded that GAA can be used in the rabbit diet to improve growth and carcass traits, and these are related to the high levels of muscle adenine nucleotide.

Guanidinoacetic acid in human nutrition: Beyond creatine synthesis.

Guanidinoacetic acid (GAA) is a nutrient that has been used in human nutrition since the early 1950s. Recommended for its role in creatine biosynthesis, GAA demonstrated beneficial energy-boosting effects in various clinical conditions. Dietary GAA has also been suggested to trigger several creatine-independent mechanisms. Besides acting as a direct precursor of high-energy phosphagen creatine, dietary GAA is suggested to reduce blood glucose concentration by acting as an insulinotropic food compound, spare amino acid arginine for other metabolic purposes (including protein synthesis), modulate taste, and perhaps alter methylation and fat deposition in various organs including the liver. GAA as a food component can have several important metabolic roles beyond creatine biosynthesis; future studies are highly warranted to address GAA overall role in human nutrition.

Effects of feeding guanidinoacetic acid on oxidative status and creatine metabolism in broilers subjected to chronic cyclic heat stress in the finisher phase.

Dietary guanidinoacetic acid (GAA) has been shown to affect creatine (Cr) metabolic pathways resulting in increased cellular Cr and hitherto broiler performances. Yet, the impact of dietary GAA on improving markers of oxidative status remains equivocal. A model of chronic cyclic heat stress, known to inflict oxidative stress, was employed to test the hypothesis that GAA could modify bird's oxidative status. A total of 720-day-old male Ross 308 broilers were allocated to 3 treatments: 0, 0.6 or 1.2 g/kg GAA was added to corn-SBM diets and fed for 39 d, with 12 replicates (20 birds each) per treatment. The chronic cyclic heat stress model (34°C with 50-60% RH for 7 h daily) was applied in the finisher phase (d 25-39). Samples from 1 bird per pen were taken on d 26 (acute heat stress) and d 39 (chronic heat stress). GAA and Cr in plasma were linearly increased by feeding GAA on either sampling day, illustrating efficient absorption and methylation, respectively. Energy metabolism in breast and heart muscle was greatly supported as visible by increased Cr and phosphocreatine: ATP, thus providing higher capacity for rapid ATP generation in cells. Glycogen stores in breast muscle were linearly elevated by incremental GAA, on d 26 only. More Cr seems to be directed to heart muscle as opposed to skeletal muscle during chronic heat stress as tissue Cr was higher in heart but lower in breast muscle on d 39 as opposed to d 26. The lipid peroxidation marker malondialdehyde, and the antioxidant enzymes superoxide dismutase and glutathione peroxidase showed no alterations by dietary GAA in plasma. Opposite to that, superoxide dismutase activity in breast muscle was linearly lowered when feeding GAA (trend on d 26, effect on d 39). Significant correlations between the assessed parameters and GAA inclusion were identified on d 26 and d 39 using principal component analysis. To conclude, beneficial performance in heat-stressed broilers by GAA is associated with enhanced muscle energy metabolism which indirectly may also support tolerance against oxidative stress.

Effects of arginine and guanidinoacetic acid with or without phenylalanine on ascites susceptibility in cold-stressed broilers fed canola meal-based diet.

In order to evaluate the effects of ARG sources (arginine [ARG] and Guanidinoacetic acid [GAA]) and phenylalanine (PHE) supplementation on performance, susceptibility to ascites, intestinal morphology, and nutrient digestibility in the cold-stressed broilers fed a canola meal (CM)-based diet, a 2×2 factorial experiment with four treatments was conducted. The dietary treatments included CM-based diet + 2.57 g/kg ARG, CM-based diet + 2.57 g/kg ARG + 1.5 g/kg PHE, CM-based diet + 1.8 g/kg GAA and CM-based diet + 1.8 g/kg GAA + 1.5 g/kg PHE. The corn-CM diet without supplementation was used as a negative control (NC) group in the fifth treatment that excluded the factorial arrangement. The results showed that adding ARG to diets without PHE supplement increased (p < 0.05) feed intake. Also, birds fed diets containing ARG had higher (p < 0.05) body weight gain (BWG) compared to those fed GAA added diets. Supplementation of PHE improved (p < 0.05) the FCR compared to groups fed diets without added PHE. Further, ARG addition increased (p < 0.05) plasma nitric oxide (NO) concentration, carcass, breast and leg yields, duodenal, jejunal, and ileal villus height (VH) to crypt depth (CD, and dry matter digestibility, while decreasing (p < 0.05) ascites mortality and right ventricle (RV) to total ventricle (TV) ratio compared to GAA added groups. Supplementation of PHE also declined susceptibility to ascites by reducing (p < 0.01) RV to TV ratio while increasing (p < 0.05) plasma NO level. The digestibility of ether extract also increased (p < 0.05) in broilers fed GAA supplemented diets versus those fed ARG added diets. The findings suggested that ARG may improve BWG and lower ascites incidence in broilers fed a diet based on CM under cold stress because of its antihypertensive effects. Moreover, the findings of this study demonstrated the importance of including PHE formulation in ARG-deficient diets to attenuate the adverse effects of cold stress on broilers. It was also concluded that GAA could be efficaciously used in cold-stressed broilers fed an ARG-deficient diet.

Dietary creatine and guanidinoacetic acid supplementation have limited effects on hybrid striped bass.

The effects of dietary supplementation of creatine and guanidinoacetic acid (GDA) have been studied to a limited extent in various fish species including red drum (Sciaenops ocellatus) and hybrid striped bass (HBS) (Morone saxatilis x M. chrysops). However, in HSB, there is a need to better understand the impact of creatine and GDA supplementation at elevated salinity which may be encountered by this euryhaline fish. Therefore, two separate feeding trials were conducted at a salinity ranging from 15 to 20 g/L with juvenile HSB for 9 and 8 weeks to evaluate the effects of dietary creatine and GDA. In each trial, four diets were formulated with either singular additions of creatine at 2% of dry weight, GDA at 1% of dry weight, or a combination of both. Fish grew adequately in both feeding trials but no significant (P > 0.05) effects of supplemental creatine or GDA were observed on weight gain, feed efficiency, survival, hepatosomatic index (HSI), intraperitoneal fat (IPF ratio), or protein conversion efficiency (PCE). However, fish fed diets supplemented with creatine had significantly (P < 0.05) increased ash and reduced lipid deposition in whole-body tissues in the first feeding trial. Supplemental creatine also resulted in significantly higher muscle yield in the second trial, but no other effects on growth performance or body composition were observed. The addition of GDA to the diet had little effect except for significantly increasing the creatine content in the liver of fish in both feeding trials due to its role as a precursor and a catalyst for synthesis of creatine within the body. Based on the results of these two trials, supplemental creatine and GDA had rather limited effects on HSB cultured in moderately saline water.

Guanidinoacetic acid supplementation improves intestinal morphology, mucosal barrier function of broilers subjected to chronic heat stress.

The current study is designed to investigate dietary guanidinoacetic acid (GAA) supplementation on the growth performance, intestinal histomorphology, and jejunum mucosal barrier function of broilers that are subjected to chronic heat stress (HS). A total of 192 male broilers (28-d old) were randomly allocated to four groups. A chronic HS model (at a temperature of 32 °C and 50%-60% relative humidity for 24 h daily) was applied in the experiment. Normal control (NC, ad libitum feeding, 22 °C), HS group (HS, ad libitum feeding, 32 °C), pair-fed group (PF, received food equivalent to that consumed by the HS group on the previous day, 22 °C), guanidinoacetic acid group (HG, ad libitum feeding, supplementing the basal diet with 0.6 g/kg GAA, 32 °C). The experiment lasted from 28 to 35 and 28 to 42 d of age of broilers. Our results showed that broilers subjected to HS had lower average daily feed intake and average daily gain (P < 0.05), higher feed-to-gain ratio and relative length of the small intestine (P < 0.05), as well as lower relative weight and weight per unit length of the small intestine (P < 0.05). HS damaged the small intestinal histomorphology by decreasing the small intestinal VH and the VH/CD (P < 0.05). Compared with the HS group, supplementation with 0.6 g/kg GAA increased jejunal VH and VH/CD (P < 0.05), but decreased relative weight and relative length of the small intestine (P < 0.05). Moreover, in comparison with NC, HS elevated intestinal permeability (D-Lactic acid concentration and diamine oxidase activity) and mRNA expression levels of interleukin-1ß, interleukin-6, and tumor necrosis factor-α (P < 0.05), reduced jejunal mucus thickness, number of goblet cells, IgA + cell density, and mucin2 mRNA expression level of broilers (P < 0.05). Compared with the HS group, dietary GAA elevated jejunal mucus thickness, goblet cell number and IgA+ cell density (P < 0.05), and up-regulated jejunal mRNA expression of interleukin-1ß and tumor necrosis factor-α (P < 0.05). In conclusion, HS impaired growth performance, and the intestinal mucosal barrier function of broilers. Dietary supplementation with 0.6 g/kg GAA alleviated HS-induced histomorphology changes of small intestine and jejunal mucosal barrier dysfunction.

With the global warming getting worse, heat stress (HS) has been a serious problem faced by poultry industry. As one of the main target organs, the intestine is easily affected by HS. Broilers are particularly sensitive to hot temperatures, and HS occurs when temperatures rise above the optimum (16­26 °C). Moreover, ambient humidity below 40% and above 80% also affects broilers adversely. HS can impair intestinal morphology and function of the intestinal barrier. The intestinal mucosal barrier not only plays key roles in nutrient digestion and absorption but also serves as the innate defense barrier fending off noxious substances within the intestinal luminal environment. Therefore, protecting intestinal mucosal barrier from HS is important to animal health. Nutrient regulation is an economical and effective method to alleviate HS of intensively-farmed broiler chickens. The results of current study demonstrated that chronic HS impaired the growth performance and intestinal mucosal barrier function of broilers, while dietary supplementation with 0.6 g/kg guanidinoacetic acid improved intestinal histomorphology and alleviated intestinal barrier dysfunction of broilers subjected to chronic HS, which is beneficial for improving health of broilers.

Effects of guanidinoacetic acid supplementation on lactation performance, nutrient digestion and rumen fermentation in Holstein dairy cows.

BACKGROUND: Considering the high energy demand of lactation and the potential of guanidinoacetic acid (GAA) addition on the increase in creatine supply for cows, the present study investigated the effects of 0, 0.3, 0.6 and 0.9 g kg-1 dry matter (DM) of GAA supplementation on lactation performance, nutrient digestion and ruminal fermentation in dairy cows. The study used 40 mid-lactation multiparous Holstein cows and the study duration was 100 days. RESULTS: DM intake was not affected, but milk and milk component yields and feed efficiency increased linearly with increasing GAA addition. The total-tract digestibility of DM, organic matter, neutral detergent fibre, acid detergent fibre and non-fibre carbohydrates increased linearly and that of crude protein increased quadratically with increasing GAA addition. When the addition level of GAA increased, ruminal pH, molar percentages of butyrate, isobutyrate and isovalerate and the acetate-to-propionate ratio decreased linearly, and the total volatile fatty acids concentration and propionate molar percentage also increased linearly, whereas the acetate molar percentage and ammonia-N concentration were unaltered. The activities of fibrolytic enzymes, α-amylase and protease increased linearly. The populations of total bacteria, fungi, Ruminococcus albus, Fibrobacter succinogenes, Ruminococcus flavefaciens, Ruminobacter amylophilus and Prevotella ruminicola increased linearly, whereas protozoa and methanogens decreased linearly with increasing GAA addition. As for the blood metabolites, concentrations of glucose, urea nitrogen and methionine were unchanged, total protein, albumin, creatine and homocysteine increased linearly, and folate decreased linearly with increasing GAA supply. CONCLUSION: The results of the present study indicate that supplementation of GAA improved milk performance and rumen fermentation in lactating dairy cows. © 2022 Society of Chemical Industry.

Serum metabolomics reveals the effects of accompanying treatment on fatigue in patients with multiple myeloma.

PURPOSE: The renewal and iteration of chemotherapy drugs have resulted in more frequent long-term remissions for patients with multiple myeloma (MM). MM has transformed into a chronic illness for many patients, but the cancer-related fatigue (CRF) of many MM convalescent patients experience is frequently overlooked. We investigated whether the accompanying treatment of family members would affect MM patients' CRF and explore their serum metabolomics, so as to provide clinicians with new ideas for identifying and treating CRF of MM patients. METHODS: This was a single-center study, and a total of 30 MM patients were included in the study. Patients were divided into two groups based on whether they have close family members accompanying them through the whole hospitalization treatment. These patients received regular chemotherapy by hematology specialists, and long-term follow-up was done by general practitioners. Patients' CRF assessment for several factors used the Chinese version of the Brief Fatigue Inventory (BFI-C). Face-to-face questionnaires were administered at a time jointly determined by the patient and the investigator. All questionnaires were conducted by a general practitioner. The LC-MS-based metabolomics analysis determined whether the patients' serum metabolites were related to their fatigue severity. A correlation analysis investigated the relationship between serum metabolites and clinical laboratory indicators. RESULTS: The fatigue severity of MM patients whose family members participated in the treatment process (group A) was significantly lower than patients whose family members did not participate in the treatment process (group B). There was a statistically significant difference (fatigue severity composite score: t = - 2.729, p = 0.011; fatigue interference composite score: t = - 3.595, p = 0.001). There were no differences between the two groups of patients' gender, age, regarding clinical staging, tumor burden, blood routine, biochemical, or coagulation indexes. There were 11 metabolites, including guanidine acetic acid (GAA), 1-(Methylthio)-1-hexanethiol, isoeucyl-asparagine, L-agaritine, tryptophyl-tyrosine, and betaine, which significantly distinguished the two groups of MM patients. GAA had the strongest correlation with patient fatigue, and the difference was statistically significant (fatigue severity composite score: r = 0.505, p = 0.0044; fatigue interference composite score: r = 0.576, p = 0.0009). The results showed that GAA negatively correlated with albumin (r = - 0.4151, p = 0.0226) and GGT (r = - 0.3766, p = 0.0402). Meanwhile, GAA positively correlated with PT (r = 0.385, p = 0.0473), and the difference was statistically significant. CONCLUSION: The study is the first to report that family presence throughout the whole hospitalization may alleviate CRF in MM patients. Moreover, the study evaluated serum metabolites linked to CRF in MM patients and found that CRF has a significant positive correlation with GAA. GAA may be a more sensitive biomarker than liver enzymes, PT, and serum albumin in predicting patient fatigue. While our sample may not represent all MM patients, it proposes a new entry point to help clinicians better identify and treat CRF in MM patients.

Dietary guanidinoacetic acid supplementation improves water holding capacity and lowers free amino acid concentration of fresh meat in finishing pigs fed with various dietary protein levels.

The current study was carried out to detect the effect of dietary guanidinoacetic acid (GAA) supplementation on carcass characteristics and meat quality in finishing pigs fed different dietary crude protein (CP) levels. Sixty-four barrows with an initial body weight of 73.05 ± 2.34 kg were randomly allocated into 1 of 4 dietary treatments in a 2 (100% vs. 125% NRC CP level) × 2 (0 vs. 300 mg/kg GAA) factorial arrangement (n = 7). The feeding trial lasted for 49 d. GAA supplementation significantly reduced drip loss (P = 0.01), free water distribution (T 23 peak area ratio) (P = 0.05) and the concentrations of free alanine, threonine, methionine and isoleucine (P < 0.05); but increased total glycine content (P = 0.03) in the longissimus dorsi muscle of finishing pigs regardless of the dietary CP levels. Furthermore, primary myogenic cell differentiation system was employed to investigate the influence of inclusion of GAA on free amino acid concentrations in myotubes (n = 4) and validate the finding in the animal feeding trial. We found that GAA inclusion in culture medium also decreased intracellular concentrations of free alanine, threonine, methionine, isoleucine, valine and proline in differentiated primary myogenic cells in vitro (P < 0.05). Meanwhile, relative to diets with 100% NRC CP level, the intake of diets with 125% NRC CP level improved sarcoplasmic protein solubility, increased the contents of carnosine and total free amino acids as well as flavor amino acids in the longissimus dorsi muscle and decreased backfat thickness at the 6-7th ribs in pigs (P < 0.05). In addition, we observed that the impact of dietary GAA supplementation on the last rib fat thickness, shear force, and free lysine content in the longissimus dorsi muscle was dependent on dietary CP levels (P < 0.05). Collectively, dietary GAA supplementation can reduce drip loss, decrease the concentrations of free amino acids and flavor amino acids of fresh meat independent of dietary CP levels.

Effect of guanidinoacetic acid supplementation on nitrogen retention and methionine methyl group flux in growing steers fed corn-based diets.

Six ruminally cannulated Holstein steers (256 ± 14 kg) were used in a 6 × 6 Latin square design to assess effects of guanidinoacetic acid (GAA) supplementation on N retention and methionine (Met) methyl group flux in growing cattle fed corn-based diets. Factorial treatments were two levels of Met (0 or 5 g/d) and three levels of GAA (0, 7.5, or 15 g/d) delivered by continuous abomasal infusion. Periods were 10 d in length and included 6 d of treatment adaptation, 3 d for total fecal and urine collections, and 1 d for blood sampling and flux measurements. Urinary N linearly increased (P < 0.01) with GAA supplementation and decreased (P < 0.05) with Met supplementation. Fecal N excretion was unaffected (P ≥ 0.42) by treatment. Retained N was not affected by GAA supplementation, but it was increased (P < 0.01) by Met supplementation. Use of methionine for transmethylation reactions, as well remethylation of homocysteine, was not affected by GAA supplementation when Met was not supplemented, but tended to be linearly increased by GAA supplementation when Met was supplemented (GAA-linear × Met interaction; P = 0.07), with the increases matching the amount of GAA provided. This response suggests that methylation reactions for compounds other than GAA were reduced by GAA supplementation when Met supply was deficient. Plasma concentrations and urinary excretion of creatine increased linearly (P = 0.03 and P = 0.06, respectively) when GAA was supplemented. There was a linear increase (P < 0.01) in urinary GAA excretion with GAA supplementation. Neither plasma concentration nor urinary excretion of creatinine was affected (P ≥ 0.17) by treatment. No treatment differences (P ≥ 0.13) were observed for plasma haptoglobin concentrations. Plasma urea-N linearly increased (P < 0.05) with GAA supplementation. Concentrations of Met and taurine increased (P < 0.05) when Met was supplemented. Plasma arginine was greatest at the intermediate level of supplemental GAA (quadratic, P < 0.05). The increase in N retention when Met was supplemented demonstrates Met was limiting in the corn-based diet. Supplementation of GAA alone or with Met as a methyl donor did not increase N retention in growing steers, perhaps because creatine production was favored over protein deposition as a use for Met.

We studied the use of methionine (Met) for methylation reactions in growing cattle by supplementing Met and guanidinoacetic acid (GAA). Supplemental GAA consumes methyl groups to form creatine even when supplies of creatine are surfeit. Our Met supplementation increased dietary Met supply from slightly deficient to adequate for protein deposition. When Met was adequate, methylation of GAA to creatine was increased by GAA supplementation, but other methylation reactions appeared to be unaffected. However, when Met supply was deficient, GAA supplementation did not increase use of Met for total methylation reactions, suggesting that methylation reactions other than creatine synthesis were decreased. In response to GAA supplementation, remethylation of homocysteine increased in amounts that corresponded to the consumption of Met for methylation reactions, such that homocysteine catabolism was not increased by GAA supplementation. Under conditions of Met deficiency, but not during Met adequacy, use of Met for methylation reactions may be limited, such that GAA supplementation decreases synthesis of methylated compounds such as choline.

Guanidinoacetic acid provides superior cardioprotection to its combined use with betaine and (or) creatine in HIIT-trained rats.

This study aimed to determine how guanidinoacetic acid (GAA) or its combined administration with betaine (B) or creatine (C) influences the cardiac function, morphometric parameters, and redox status of rats subjected to high-intensity interval training (HIIT). This research was conducted on male Wistar albino rats exposed to HIIT for 4 weeks. The animals were randomly divided into five groups: HIIT, HIIT + GAA, HIIT + GAA + C, HIIT + GAA + B, and HIIT + GAA + C + B. After completing the training protocol, GAA (300 mg/kg), C (280 mg/kg), and B (300 mg/kg) were applied daily per os for 4 weeks. GAA supplementation in combination with HIIT significantly decreased the level of both systemic and cardiac prooxidants ( O 2 - , H2O2, NO 2 - , and thiobarbituric acid reactive substances) compared with nontreated HIIT (p < 0.05). Also, GAA treatment led to an increase in glutathione and superoxide dismutase levels. None of the treatment regimens altered cardiac function. A larger degree of cardiomyocyte hypertrophy was observed in the HIIT + GAA group, which was reflected through an increase of the cross-sectional area of 27% (p < 0.05) and that of the left ventricle wall thickness of 27% (p < 0.05). Since we showed that GAA in combination with HIIT may ameliorate oxidative stress and does not alter cardiac function, the present study is a basis for future research exploring the mechanisms of cardioprotection induced by this supplement in an HIIT scenario.

Skeletal muscle mass, meat quality and antioxidant status in growing lambs supplemented with guanidinoacetic acid.

Guanidinoacetic acid (GAA) exists naturally as a precursor of creatine, which possesses several biological functions. In the present study, the effects of dietary GAA supplementation on skeletal muscle mass and meat quality of lambs were investigated. The GAA supplementation increased final body weight, promoted muscle mass and changed the distribution of myofiber size. Meanwhile, elevated ultimate pH and water holding capacity (WHC) of resulting meat were observed in GAA fed lambs. Moreover, the total antioxidative capacity was elevated. Dietary GAA accelerated myofibril protein synthesis through regulation with IGF-1/Akt/mTOR signaling pathway and minimized protein breakdown via regulating abundances of myostatin and phosphorylated FoxO1. In vitro, GAA treatment inhibited sheep primary myoblasts proliferation, and enhanced its myogenic potential. Collectively, these results suggested that GAA might be a feed additive for use by the lamb meat industry as it has potential to improve growth performance, antioxidant status and WHC of resulting meat.