Dietary l-arginine supplementation alleviates immunosuppression induced by cyclophosphamide in weaned pigs

A study was conducted to investigate the effects of L-arginine (Arg) on performance and immune function in cyclophosphamide (CY) immunosuppressed weaned pigs. The weaned pigs were allotted randomly into one of three treatments, including: (1) non-challenged control; (2) CY-challenged group; and (3) CY + 0.5% Arg. On days 14 and 21 of the trial, pigs were injected with CY or sterile saline. Blood samples were obtained on days 21 and 28 of the trial for further analysis. On day 28, delayed-type hypersensitivity reaction was evaluated. Arg alleviated the decrease of average daily gain (P < 0.05) induced by CY challenge from days 21 to 28. Arg mitigated the CY-induced decrease of total white blood cell numbers (P < 0.05) on day 28 and improved the lymphocyte percentage on day 21 (P < 0.05). Arg increased the delayed-type hypersensitivity reaction (P < 0.05), and attenuated the decrease of bovine serum albumin antibody level caused by CY treatment (P < 0.05) on day 28. In addition, Arg elevated the levels of serum interleukin-2 and interferon-gamma (P < 0.05) on day 28, and mitigated the decrease of serum interferon-gamma level on day 21 (P < 0.05). These results indicate that Arg supplementation has beneficial effects in attenuating the immunosuppressive effects of CY challenge, therefore improving growth performance of young pigs.     

Dietary l-arginine supplementation differentially regulates expression of lipid-metabolic genes in porcine adipose tissue and skeletal muscle

Obesity is a major health crisis worldwide and new treatments are needed to fight this epidemic. Using the swine model, we recently reported that dietary l-arginine (Arg) supplementation promotes muscle gain and reduces body-fat accretion. The present study tested the hypothesis that Arg regulates expression of key genes involved in lipid metabolism in skeletal muscle and white adipose tissue. Sixteen 110-day-old barrows were fed for 60 days a corn- and soybean-meal-based diet supplemented with 1.0% Arg or 2.05% l-alanine (isonitrogenous control). Blood samples, longissimus dorsi muscle and overlying subcutaneous adipose tissue were obtained from 170-day-old pigs for biochemical studies. Serum concentrations of leptin, alanine and glutamine were lower, but those for Arg and proline were higher in Arg-supplemented pigs than in control pigs. The percentage of oleic acid was higher but that of stearic acid and linoleic acid was lower in muscle of Arg-supplemented pigs, compared with control pigs. Dietary Arg supplementation increased mRNA levels for fatty acid synthase in muscle, while decreasing those for lipoprotein lipase, glucose transporter-4, and acetyl-coenzyme A carboxylase-α in adipose tissue. Additionally, mRNA levels for hormone sensitive lipase were higher in adipose tissue of Arg-supplemented pigs compared with control pigs. These results indicate that Arg differentially regulates expression of fat-metabolic genes in skeletal muscle and white adipose tissue, therefore favoring lipogenesis in muscle but lipolysis in adipose tissue. Our novel findings provide a biochemical basis for explaining the beneficial effect of Arg in improving the metabolic profile in mammals (including obese humans).     

Dietary l-arginine supplementation enhances placental growth and reproductive performance in sows

Suboptimal embryonic/fetal survival and growth remains a significant problem in mammals. Using a swine model, we tested the hypothesis that dietary l-arginine supplementation during gestation may improve pregnancy outcomes through enhancing placental growth and modulating hormonal secretions. Gestating pigs (Yorkshire?×?Landrace, n?=?108) were assigned randomly into two groups based on parity and body weight, representing dietary supplementation with 1.0% l-arginine–HCl or 1.7% l-alanine (isonitrogenous control) between days 22 and 114 of gestation. Blood samples were obtained from the ear vein on days 22, 40, 70 and 90 of gestation. On days 40, 70 and 90 of gestation, concentrations of estradiol in plasma were higher (P?<?0.05) in arginine-supplemented than in control sows. Moreover, arginine supplementation increased (P?<?0.05) the concentrations of arginine, proline and ornithine in plasma, but concentrations of urea or progesterone in plasma did not differ between the two groups of sows. Compared with the control, arginine supplementation increased (P?<?0.05) the total number of piglets by 1.31 per litter, the number of live-born piglets by 1.10 per litter, the litter birth weight for all piglets by 1.36?kg, and the litter birth weight for live-born piglets by 1.70?kg. Furthermore, arginine supplementation enhanced (P?<?0.05) placental weight by 16.2%. The weaning-to-estrus interval of sows was not affected by arginine supplementation during gestation. These results indicate that dietary arginine supplementation beneficially enhances placental growth and the reproductive performance of sows.     

Dietary l-arginine supplementation enhances the immune status in early-weaned piglets

This study was conducted to test the hypothesis that dietary L-arginine supplementation enhances immunity in early weaned piglets. Seventy piglets weaned at 7 days of age were assigned to five groups (14 pigs/group), representing supplementation of 0.0, 0.2, 0.4, 0.6, and 0.8% L-arginine to a milk-based formula. On Day 7 after initiation of treatment, spleen weight in piglets supplemented with 0.2 and 0.8% arginine was heavier and thymus size was larger in piglets supplemented with 0.6% arginine, whereas serum concentration of immunoglobulin (Ig) M was higher but that of IL-8 was lower in piglets supplemented with 0.6 and 0.8% arginine, compared with the control group. Dietary supplementation with 0.8% arginine increased the numbers of white blood cells and granulocytes, and gene expression of interleukin (IL)-8 in spleen. On Day 14, compared with control piglets, granulocyte numbers were greater but lymphocyte numbers were lower in piglets supplemented with 0.2 and 0.4% arginine, whereas splenic expression of IL-8 and tumor necrosis factor-alpha genes was increased in piglets supplemented with 0.8% arginine. Additionally, IgG and IgM concentrations in serum and growth performance were greater in piglets supplemented with 0.4-0.8% arginine, compared with unsupplemented piglets. Collectively, dietary supplementation with 0.4-0.8% L-arginine for 2 weeks enhances both cellular and humoral immunity in piglets by modulating the production of leukocytes, cytokines and antibodies. These results indicate that increasing L-arginine provision is beneficial for optimal immune responses in young pigs and also have important implications for designing the next generation of improved formula for human infants.     

Dietary calcium attenuation of body fat gain during high-fat feeding in mice

Human epidemiological studies have supported the hypothesis that a dairy food-rich diet is associated with lower fat accumulation, although prospective studies and intervention trials are not so conclusive and contradictory data exist in animal models. The purpose of this study was to assess the effects on body weight and fat depots of dairy calcium (12 g/kg diet) in wild-type mice under ad libitum high-fat (43%) and normal-fat (12%) diets and to gain comprehension on the underlying mechanism of dairy calcium effects. Our results show that calcium intake decreases body weight and body fat depot gain under high-fat diet and accelerates weight loss under normal-fat diet, without differences in food intake. No differences in gene or protein expression of UCP1 in brown adipose tissue or UCP2 in white adipose tissue were found that could be related with calcium feeding, suggesting that calcium intake contributed to modulate body weight in wild-type mice by a mechanism that is not associated with activation of brown adipose tissue thermogenesis. UCP3 protein but not gene expression increased in muscle due to calcium feeding. In white adipose tissue there were effects of calcium intake decreasing the expression of proteins related to calcium signalling, in particular of stanniocalcin 2. CaSR levels could play a role in decreasing cytosolic calcium in adipocytes and, therefore, contribute to the diminution of fat accretion. Results support the anti-obesity effect of dietary calcium in male mice and indicate that, at least at the time-point studied, activation of thermogenesis is not involved.     

Dietary conjugated linoleic acid reduces body fat mass and affects gene expression of proteins regulating energy metabolism in mice

ICR and C57BL/6J mice were fed experimental diets containing either a 2% fatty acid preparation rich in conjugated linoleic acid (CLA) or a preparation rich in linoleic acid and free of CLA for 21 days. CLA greatly decreased weights of white adipose tissue and interscapular brown adipose tissue in the two strains. CLA reduced mRNA levels of glucose transporter 4 (Glut 4) in white and brown adipose tissue of both strains. A CLA-dependent decrease in mRNA levels of peroxisome proliferator activated receptor (PPAR) gamma was seen in interscapular brown adipose tissue of both strains and in white adipose tissue of C57BL/6J but not ICR mice. Dietary CLA was found to cause a decrease in the mRNA levels of uncoupling protein (UCP) 1 in brown adipose tissue when the value was corrected for the expression of a house-keeping gene (beta-actin) in the two strains. Uncorrected values were, however, indistinguishable between the animals fed the CLA diet and CLA-free diet. UCP 3 expression in brown adipose tissue was much lower in mice fed the CLA diet than in those fed the control diet in both strains. In contrast, CLA greatly up-regulated the gene expression of UCP 2 in brown adipose tissue. Dietary CLA also increased UCP 2 mRNA level in skeletal muscle. It is apparent that dietary CLA decreases white and brown adipose tissue mass, accompanying changes in the gene expression of proteins regulating energy metabolism in white and brown adipose tissues, and skeletal muscle of mice.     

Dynorphin knockout reduces fat mass and increases weight loss during fasting in mice

Endogenous opioids, particularly dynorphins, have been implicated in regulation of energy balance, but it is not known how they mediate this in vivo. We investigated energy homeostasis in dynorphin knockout mice (Dyn(-/-) mice) and probed the interactions between dynorphins and the neuropeptide Y (NPY) system. Dyn(-/-) mice were no different from wild types with regards to body weight and basal and fasting-induced food intake, but fecal output was increased, suggesting decreased nutrient absorption, and they had significantly less white fat and lost more weight during a 24-h fast. The neuroendocrine and thermal responses to fasting were at least as pronounced in Dyn(-/-) as in wild types, and there was no stimulatory effect of dynorphin knockout on 24-h energy expenditure (kilocalories of heat produced) or physical activity. However, Dyn(-/-) mice showed increased circulating concentrations of 3,4-dihydroxyphenlacetic acid and 3,4-dihydroxyphenylglycol, suggesting increased activity of the sympathetic nervous system. The respiratory exchange ratio of male but not female Dyn(-/-) mice was reduced, demonstrating increased fat oxidation. Interestingly, expression of the orexigenic acting NPY in the hypothalamic arcuate nucleus was reduced in Dyn(-/-) mice. However, fasting-induced increases in pre-prodynorphin expression in the arcuate nucleus, the paraventricular nucleus, and the ventromedial hypothalamus but not the lateral hypothalamus were abolished by deletion of Y(1) but not Y(2) receptors. Therefore, ablation of dynorphins results in increases in fatty acid oxidation in male mice, reductions in adiposity, and increased weight loss during fasting, possibly via increases in sympathetic activity, decreases in intestinal nutrient absorption, and interactions with the NPYergic system.     

Dietary L-arginine supplementation reduces abdominal fat content by modulating lipid metabolism in broiler chickens

This study investigated the effects of different levels of dietary L-arginine (L-Arg) supplementation on the abdominal fat pad, circulating lipids, hepatic fatty acid synthase (FAS) gene expression, gene expression related to fatty acid β-oxidation, and the performance of broiler chickens. We tested whether the dietary L-Arg levels affected the expression of genes related to lipid metabolism in order to reduce body fat deposition. A total of 192 broiler chickens (Cobb 500) aged 21 days with an average BW of 920 ± 15 g were randomly assigned to four groups (six broilers per replicate and eight replicates per treatment). The control group was fed a basal diet, whereas the treatment groups were fed basal diets supplemented with 0.25%, 0.50%, or 1.00% L-Arg for 3 weeks. The average daily feed intake, average daily gain and feed : gain ratio were not affected by the dietary L-Arg levels. However, chickens supplemented with L-Arg had lower abdominal fat content, plasma triglyceride (TG), total cholesterol (TC) concentrations, hepatic FAS mRNA expression and increased heart carnitine palmitoyl transferase1 (CPT1) and 3-hydroxyacyl-CoA dehydrogenase (3HADH) mRNA expression. These findings suggest that the addition of 0.25% L-Arg may reduce the plasma TC concentration by decreasing hepatic 3-hydroxyl-3-methylglutaryl-CoA reductase mRNA expression. This may lower the plasma TG and abdominal fat content by suppressing hepatic FAS mRNA expression and enhancing CPT1 and 3HADH (genes related to fatty acid β-oxidation) mRNA expression in the hearts of broiler chickens.     

Soy protein isolate in the presence of cornstarch reduces body fat gain in rats

The objective of the present study was to determine the combined effects of dietary protein and carbohydrate sources on total body energy and protein and fat gains as well as on plasma insulin and glucose and tissue lipoprotein lipase activity in male Sprague-Dawley rats fed semipurified diets for 28 days. The diets varied in both protein and carbohydrate sources, namely, casein-cornstarch, casein-sucrose, soy protein isolate (SPI)-cornstarch, SPI-sucrose, cod protein-cornstarch, and cod protein-sucrose. When SPI was combined with cornstarch, lower total body energy and fat gains were observed compared with the combination of either casein and sucrose, casein and cornstarch, or SPI and sucrose. Plasma glucose and insulin concentrations in addition to total and metabolizable energy intake and body weight gain were lower in rats fed the SPI-cornstarch diet than in those fed the casein-sucrose diet. Feeding the SPI-cornstarch diet compared with feeding either the casein-cornstarch or the SPI-sucrose diet also caused lower plasma glucose concentrations and a concomitant trend (p = 0.06) to reduced energy intake and body weight gain. Therefore, the reducing effects of the SPI-cornstarch diet compared with the casein-cornstarch, the casein-sucrose, and the SPI-sucrose diets on body energy and fat gains may result from reductions in energy intake and in plasma glucose concentrations.     

Age variations in the relation of body mass indices to estimates of body fat and muscle mass

In some chronic disease studies, distinctions have been made regarding the importance of body mass index (BMI) as a risk factor in younger versus older men and women. In order to determine the significance of these differences in BMI-disease associations, we determined the extent of age-dependent variations in the relation of BMIs to body composition in large probability samples of U.S. men and women from the First and Second U.S. National Health and Nutrition Examination Surveys (NHANES I and II). BMIs are more highly correlated with estimates of body fat in younger than in older men and women, and with muscle mass in older than in younger adults. Caution should be exercised in interpreting the significance of BMI as a risk factor for chronic disease, particularly in comparison of age groups.     

Neonatal dietary supplementation of arachidonic acid increases prostaglandin levels in adipose tissue but does not promote fat mass development in guinea pigs

The role of arachidonic acid (AA) on the development of adipose tissue is still controversial since its metabolites, i.e., prostaglandins, can either stimulate or inhibit preadipocyte differentiation in vitro. In the present study, we evaluated the effects of early postnatal supplementation of AA on body weight and adipose tissue development in guinea pigs. Male newborn guinea pigs were fed for 21 days (day 21) with diets (milk and pellet) supplemented (+AA) or not (-AA) with 1.2% (total fatty acids) AA. From day 21 to day 105 both groups were fed a chow diet. The 21-days-old +AA pups showed a twofold higher AA accretion in phospholipids associated with a two- to sixfold increase in several prostaglandins, such as 6-keto PGF(1alpha) (the stable hydrolysis product of PGI(2)), PGF(2alpha), PGE(2), and PGD(2) in adipose tissue, compared with the -AA group. No difference in fat pad and body weight, aP2, and leptin gene expression in adipose tissue, fasting plasma glucose, free-fatty acids, and triglyceride concentration was observed between groups at day 21 or day 105. These results show that dietary supplementation of AA during the suckling/weaning period increases prostaglandin levels in adipose tissue but does not influence early fat mass development in the guinea pig.     

Dietary tyrosine supplementation enhances weight gain in streptozotocin-diabetic rats

The effect of dietary protein concentration and tyrosine supplementation on growth in streptozotocin (65 mg/kg, ip) diabetic rats was evaluated. When rats were fed diets ranging from 15 to 60% protein, weight gain and feed efficiency were greatest in rats fed the 45% protein diet. Adding tyrosine to this diet (8%, incorporated as a percentage of protein) did not promote further weight gain relative to nonsupplemented diabetic animals. In contrast, rats choosing 45% of total calories as protein by selecting from 10 and 60% protein diets supplemented with either 0, 4, or 8% tyrosine demonstrated a 35% (4% tyrosine) to 45% (8% tyrosine) increase in weight gain. Proximate analysis indicated similar body composition in tyrosine supplemented and nonsupplemented diabetic animals. Including tryptophan (1.45%) with tyrosine in the self-selection diet was without effect. Thus, tyrosine supplementation promoted a modest but consistent and specific increase in weight gained by self-selecting diabetic rats.     

Evaluation of energy systems in determining the energy cost of gain of growing-finishing pigs fed diets containing different levels of dietary fat

Two experiments were conducted to evaluate the impact of different energy systems in predicting the energy cost of gain of growing-finishing pigs fed diets containing different levels of dietary fat. The diets in both experiments were based on wheat, corn and soybean meal and supplemented with 0, 1.75, 3.50 and 5.25% tallow. In Experiment 1, 24 crossbred barrows (Duroc x Landrace x Yorkshire) were randomly allocated to one of the four dietary treatments to determine the digestible (DE) and metabolizable (ME) energy content of the diets and net energy (NE) was calculated from DE and ME values. In Experiment 2, 96 crossbred barrows (Duroc x Landrace x Yorkshire) were used to evaluate the effectiveness of the energy systems in predicting the energy cost of gain for growing-finishing pigs fed ad libitum. There were six pens per treatment and four pigs per pen. The results obtained in trial 1 were used for calculating the energy cost of gain in trial 2. During the growing period, there was a linear decrease (p < 0.05) in the DE and ME cost of gain, while the NE cost of gain was not influenced by level of fat. During the finishing period, neither DE, ME or NE cost of gain were influenced by the dietary fat level. For the total experiment, the DE and ME cost of gain decreased (linear effect; p = 0.001), but there was no significant decrease in the NE cost of gain. It is concluded that the NE system can predict the performance of growing-finishing pigs more precisely for diets differing in fat content than can the DE and ME systems.     

The transgenic expression of human follistatin-344 increases skeletal muscle mass in pigs

Follistatin (FST), which was first found in the follicles of cattle and pigs, has been shown to be an essential regulator for muscle development. Mice that were genetically engineered to overexpress Fst specifically in muscle had at least twice the amount of skeletal muscle mass as controls; these findings are similar to earlier results obtained in myostatin-knockout mice. However, the role of follistatin in skeletal muscle development has yet to be clarified in livestock. Here, we describe transgenic Duroc pigs that exogenously express Fst specifically in muscle tissue. The transgenic pigs exhibited an increased proportion of skeletal muscle and a reduced proportion of body fat that were similar to those reported in myostatin-null cattle. The lean percentage of lean meat was significantly higher in the F1 generation of TG pigs (72.95 ± 1.0 %) than in WT pigs (69.18 ± 0.97 %) (N = 16, P < 0.05). Myofiber hypertrophy was also observed in the longissimus dorsi of transgenic pigs, possibly contributing to the increased skeletal muscle mass. Western blot analysis showed a significantly reduced level of Smad2 phosphorylation and an increased level of Akt^S473 phosphorylation in the skeletal muscle tissue of the transgenic pigs. Moreover, no cardiac muscle hypertrophy or reproductive abnormality was observed. These findings indicate that muscle-specific Fst overexpression in pigs enhances skeletal muscle growth, at least partly due to myofiber hypertrophy and providing a promising approach to increase muscle mass in pigs and other livestock.     

Dietary supplementation with l-arginine partially counteracts serum metabonome induced by weaning stress in piglets

Arginine plays an important role in preventing intestinal dysfunction and metabolic disorders caused by early weaning stress. However, little is known about how arginine mitigates early weaning stress. This study was conducted to evaluate the effects of weaning stress and dietary arginine supplementation on the metabonome in the serum of piglets using (1)H NMR spectroscopy in conjunction with multivariate data analysis. Thirty castrated male piglets aged 21 d were evenly divided into three groups and fed in three different regimes: sow-fed (SF), weaned with l-alanine supplementation (ALA), and weaned with arginine supplementation (ARG). We found that early weaning stress led to a significantly reduced bodyweight gain (15.6%) and that supplementation with arginine can improve growth rates in piglets by 5.6% (P < 0.05). The early weaning stress was associated with marked alterations in lipid and amino acid metabolisms and perturbations in population and/or activities of gut microorganisms, which were manifested in increased levels of organic acids, amino acids, and acetyl-glycoproteins and reduced levels of choline metabolism and lipoproteins. Dietary supplementation with arginine could partially counteract the changes of metabolites induced by weaning stress, such as lipid and amino acid metabolisms. However, arginine was not able to restore disturbed gut microbiota. These results demonstrate the central role of arginine supplementation in regulating the metabolisms of weaned piglets.     

Dietary l-arginine supplementation improves semen quality and libido of boars under high ambient temperature

l-Arginine is a nutritionally essential amino acid for spermatogenesis and plays versatile roles in animal health and can be utilized as a potential agent to improve reproductive performance of boars under high ambient temperature. The present study aimed to determine whether dietary l-arginine could alleviate heat stress-induced infertility in boars. In all, 20 boars (PIC 1040; 248.59±3.84 kg BW and 407.65±6.40 days of age) were selected and randomly assigned to four groups (group 0.0%, basal diet; group 0.6%, 0.8% or 1.0%, basal diet added with 0.6%, 0.8% or 1.0% l-arginine (wt:wt), respectively.) The four diets were made isonitrogenous by addition of appropriate amounts of l-alanine. Boars were pre-fed the corresponding experimental diet for 42 days. Then, the semen characteristics and libido were accessed for 6 weeks during the hot summer period (25.5° to 33.0°C). Results show that dietary l-arginine remarkably improved sperm motility, normality, total sperm number and effective total sperm number. Also, dietary l-arginine improved semen antioxidant capacity, such as decrease of malondialdehyde and 8-Hydroxy-2'-deoxyguanosine content in sperm (P<0.05), increase of the ratio of glutathione and oxidized glutathione, total antioxidant capacity, glutathione peroxidase and catalase activities in seminal plasma (P<0.05). Most of mitochondria contained intact ultrastructure in l-arginine-supplemented group which also accompany with higher ATP content than the 0.0% group. The boars fed 0.8% l-arginine show increased levels of estradiol-17β and testosterone and exhibit improved libido performance than boars in the 0.0% group. Adding dietary l-arginine linearly increased (P=0.002) nitric oxide content (as l-arginine increased). The scrotal surface temperature in the 0.6%, 0.8% and 1.0% group were decreased by 0.9°C, 0.9°C and 0.4°C, respectively, compared with the 0.0% group. l-Arginine levels caused linear effect on semen quality and antioxidant capacity, also caused quadratic effect on libido performance. During the hot summer months, the predicted optimal l-arginine levels for best semen quality and antioxidant capacity was 0.8% to 1.0% and for best libido performance was 0.8%. It can be concluded that l-arginine can be used as an effective agent to alleviate heat stress-induced infertility of boar, and that 0.8% to 1.0% can be considered as the optimum dosage.