Dietary supplementation of laminarin improves the reproductive performance of sows and the growth of suckling piglets

Background Maternal nutrition is essential in keeping a highly efficient production system in the pig industry.Laminarin has been shown to improve antioxidant capacity,reduce the inflammatory response,and favor the homeostasis of intestinal microbiota.However,the effect of dietary supplementation of laminarin on the reproductive performance of sows and the growth of suckling offspring remains unknown.Methods A total of 40 Landrace×Yorkshire multiparous sows on d 85 of gestation,similar in age,body weight(BW),parity and reproductive performance,were randomly divided into four dietary treatments with 10 sows per treatment,receiving a control diet(basal pregnancy or lactating diets)and a basal diet supplemented with 0.025%,0.05%and 0.10%laminarin,respectively.The experiment lasted from d 85 of gestation to d 21 of lactation.Results Laminarin supplementation linearly increased number born alive per litter(P=0.03),average daily feed intake(ADFI,P<0.01),and total milk yield of sows during the lactation of 1–21 d(P=0.02).Furthermore,maternal laminarin supplementation increased the average daily gain(ADG)of piglets while tending to reduce the culling and death rate before weaning.In addition,alterations to the composition of colostrum and milk,as well as to serum inflammatory cytokines and immunoglobulins of sows were observed.The fecal microbiota profile of sows supported the improvement of reproductive performance in sows and the growth performance in suckling offspring.Conclusions Dietary supplementation of laminarin during late pregnancy and lactation could significantly improve reproductive performance of sows and growth performance of piglets.

Lysine 2-hydroxyisobutyrylation levels determined adipogenesis and fat accumulation in adipose tissue in pigs

Background Excessive backfat deposition lowering carcass grade is a major concern in the pig industry,especially in most breeds of obese type pigs.The mechanisms involved in adipogenesis and fat accumulation in pigs remain unclear.Lysine 2-hydroxyisobutyrylation(Khib),is a novel protein post-translational modification(PTM),which play an important role in transcription,energy metabolism and metastasis of cancer cells,but its role in adipogenesis and fat accumulation has not been shown.Results In this study,we first analyzed the modification levels of acetylation(Kac),Khib,crotonylation(Kcr)and succinylation(Ksu)of fibro-adipogenic progenitors(FAPs),myogenic precursors(Myo)and mesenchymal stem cells(MSCs)with varied differentiation potential,and found that only Khib modification in FAPs was significantly higher than that in MSCs.Consistently,in parallel with its regulatory enzymes lysine acetyltransferase 5(KAT5)and histone deacetylase 2(HDAC2)protein levels,the Khib levels increased quadratically(P<0.01)during adipogenic differentiation of FAPs.KAT5 knockdown in FAPs inhibited adipogenic differentiation,while HDAC2 knockdown enhanced adipogenic differentiation.We also demonstrated that Khib modification favored to adipogenic differentiation and fat accumulation by comparing Khib levels in FAPs and backfat tissues both derived from obese-type pigs(Laiwu pigs)and lean-type pigs(Duroc pigs),respectively.Accordingly,the expression patterns of KAT5 and HDAC2 matched well to the degree of backfat accumulation in obese-and lean-type pigs.Conclusions From the perspective of protein translational modification,we are the first to reveal the role of Khib in adipogenesis and fat deposition in pigs,and provided new clues for the improvement of fat accumulation and distribution as expected via genetic selection and nutritional strategy in obese-type pigs.

Insights into the mechanism of L-malic acid on drip loss of chicken meat under commercial conditions

Background A deterioration in the meat quality of broilers has attracted much more attention in recent years.L-malic acid(MA)is evidenced to decrease meat drip loss in broilers,but the underlying molecular mechanisms are still unclear.It’s also not sure whether the outputs obtained under experimental conditions can be obtained in a com-mercial condition.Here,we investigated the effects and mechanisms of dietary MA supplementation on chicken meat drip loss at large-scale rearing.Results Results showed that the growth performance and drip loss were improved by MA supplementation.Meat metabolome revealed that L-2-aminoadipic acid,β-aminoisobutyric acid,eicosapentaenoic acid,and nicotinamide,as well as amino acid metabolism pathways connected to the improvements of meat quality by MA addition.The transcriptome analysis further indicated that the effect of MA on drip loss was also related to the proper immune response,evidenced by the enhanced B cell receptor signaling pathway,NF-κB signaling pathway,TNF signaling pathway,and IL-17 signaling pathway.Conclusions We provided evidence that MA decreased chicken meat drip loss under commercial conditions.Metabolome and transcriptome revealed a comprehensive understanding of the underlying mechanisms.Together,MA could be used as a promising dietary supplement for enhancing the water-holding capacity of chicken meat.

Alfalfa-containing diets alter luminal microbiota structure and short chain fatty acid sensing in the caecal mucosa of pigs

Background: Pork produced by outdoor-reared pigs raised mostly on alfalfa pastures attracts increasing population of consumer from most of the world. In China, pigs were raised with alfalfa-containing diets to seek for good quality pork.However, the influence of dietary alfalfa involving high level of insoluble dietary fiber(IDF) on pig intestinal luminal microbiota composition remains unclear. The objective of this study was to investigate the effects of alfalfa on luminal microbiota and short chain fatty acids(SCFA) production, and gene expressions involved in SCFA sensing, transporting and absorbing in pig caecal mucosa.Results: Twenty-four growing pigs were randomly allotted to four diets containing 0%, 5%, 10% and 15% alfalfa meal for a 28-d experiment. Ingestion of alfalfa meal-contained diets significantly increased the ratio of body weight gain to feed consumption. Illumina MiS eq sequencing of the V3 region of the 16 S r RNA genes showed that alfalfa-containing diet significantly decreased the relative abundance of genera Turicibacter, Acidiphilium, Paracoccus, Propionibacterium,Corynebacterium, Pseudomonas, Acinetobacter, and Staphylococcus, and increased the relative abundance of genera Lachnospira, Marvinbryantia, and Desulfovibrio in the caecal digesta. Butyrate concentration was significantly increased in the hindgut by the supplementation of alfalfa meal in diets. The m RNA gene expressions of FFAR3, SMCT1, MCT1,PYY, and GCG were significantly increased in the caecal mucosa of pigs fed alfalfa meal.Conclusions: Our results suggested that alfalfa-containing diet has exerted significant impacts on caecal microbiota composition, butyrate concentration and significantly upregulated m RNA expression of host caecal mucosal genes involved in SCFA sensing and absorption as well as regulation of satiety.

Surplus dietary isoleucine intake enhanced monounsaturated fatty acid synthesis and fat accumulation in skeletal muscle of finishing pigs

Background: Isoleucine(Ile) has been implicated in the regulation of energy homeostasis and adipogenesis.However,the impact of surplus dietary Ile intake on muscle lipogenesis remains unknown.The present study aimed to investigate the impact of dietary supplementation of extra-Ile on lipogenesis,fatty acid profile and lipid accumulation in skeletal muscle in finishing pigs.Methods: Forty-eight barrows with initial body weight of 77.0 ± 0.1 kg were allotted to one of two groups and fed diets containing 0.39%,0.53% standardized ileal digestible(SID) Ile with six replicates per treatment and four pigs per replicate for 30 d.Results: Dietary Ile intake significantly improved the intramuscular fat(IMF) content and monounsaturated fatty acid(MUFA) concentration in the skeletal muscle(P < 0.05),and decreased the drip loss and shear force(P < 0.05) without influencing the growth performance of pigs(P > 0.05).Moreover,the phosphorylation of adenosine monophosphate activated protein kinase α(AMPKα) and acetyl coenzyme A carboxylase(ACC) proteins that monitor lipid metabolism were decreased in skeletal muscle of pigs offered extra-Ile diet(P < 0.05).The mRNA expression of adipose-specific genes adipocyte determination and differentiation factor 1(ADD1),fatty acid synthase(FAS),and stearoyl-CoA desaturase(SCD) were upregulated and the activity of SCD was increased as well(P < 0.05).Conclusions: Surplus dietary Ile intake could increase IMF accumulation and MUFA synthesis in skeletal muscle through depressing the phosphorylation of AMPKα-ACC and stimulating the expression and activity of SCD,and increasing the capability of lipogenesis in skeletal muscle.

Selenium deficiency induces spleen pathological changes in pigs by decreasing selenoprotein expression,evoking oxidative stress,and activating inflammation and apoptosis

Background:The immune system is one aspect of health that is affected by dietary selenium(Se)levels and selenoprotein expression.Spleen is an important immune organ of the body,which is directly involved in cellular immunity.However,there are limited reports on Se levels and spleen health.Therefore,this study established a Se-deficient pig model to investigate the mechanism of Se deficiency-induced splenic pathogenesis.Methods:Twenty-four pure line castrated male Yorkshire pigs(45 days old,12.50±1.32 kg,12 full-sibling pairs)were divided into two equal groups and fed Se-deficient diet(0.007 mg Se/kg)or Se-adequate diet(0.3 mg Se/kg)for 16 weeks.At the end of the trial,blood and spleen were collected to assay for erythroid parameters,the osmotic fragility of erythrocytes,the spleen index,histology,terminal deoxynucleotidyl transferase nick-end labeling(TUNEL)staining,Se concentrations,the selenogenome,redox status,and signaling related inflammation and apoptosis.Results:Dietary Se deficiency decreased the erythroid parameters and increased the number of osmotically fragile erythrocytes(P<0.05).The spleen index did not change,but hematoxylin and eosin and TUNEL staining indicated that the white pulp decreased,the red pulp increased,and splenocyte apoptosis occurred in the Se deficient group.Se deficiency decreased the Se concentration and selenoprotein expression in the spleen(P<0.05),blocked the glutathione and thioredoxin antioxidant systems,and led to redox imbalance.Se deficiency activated the NF-κB and HIF-1αtranscription factors,thus increasing pro-inflammatory cytokines(IL-1β,IL-6,IL-8,IL-17,and TNF-α),decreasing anti-inflammatory cytokines(IL-10,IL-13,and TGF-β)and increasing expression of the downstream genes COX-2 and iNOS(P<0.05),which in turn induced inflammation.In addition,Se-deficiency induced apoptosis through the mitochondrial pathway,upregulated apoptotic genes(Caspase3,Caspase8,and Bak),and downregulated antiapoptotic genes(Bcl-2)(P<0.05)at the mRNA level,thus verifying the results of TUNEL staining.Conclusions:These results indicated that Se deficiency induces spleen injury through the regulation of selenoproteins,oxidative stress,inflammation and apoptosis.

Global transcriptomic analysis reveals Lnc-ADAMTS9 exerting an essential role in myogenesis through modulating the ERK signaling pathway

Background:Long non-coding RNAs(lncRNAs)are emerging key regulators involved in a variety of biological processes such as cell differentiation and development.The balance between myogenesis and adipogenesis is crucial for skeletal muscle homeostasis in humans and meat quality in farm animals.The present study aimed to reveal the global transcriptomic profiles of adipogenic(Adi-)and myogenic(Myo-)precursors derived from porcine skeletal muscle and identify lncRNAs involved in the modulation of myogenesis homeostasis in porcine skeletal muscle.Results:In this study,a total of 655 novel individual lncRNAs including differentially expressed 24 lncRNAs,and 755 differentially expressed mRNAs were identified(fold change≥2 or≤0.5 and adjusted P<0.05).Integrated results of Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis accompanied by the variation of intracellular Ca2+concentration highlighted Lnc-ADAMTS9 involved in the modulation of myogenesis homeostasis in porcine skeletal muscle.Although Lnc-ADAMTS9 knock-down did not alter the mRNA expression of ADAMTS9,we demonstrated that Lnc-ADAMTS9 can promote myogenic proliferation and myogenic differentiation of myogenic precursors through inhibiting the ERK/MAPK signaling pathway.Conclusion:We deciphered a comprehensive catalog of mRNAs and lncRNAs that might be involved in the regulation of myogenesis and adipogenesis homeostasis in the skeletal muscle of pigs.The Lnc-ADAMTS9 exerts an essential role in myogenesis through the ERK signaling pathway.

A systematic review and meta-analysis of dietary fat effects on reproductive performance of sows and growth performance of piglets

Effects of added fat in sow diets on the sows’reproductive performance and offspring growth performance are influenced by multiple factors such as genetics,nutrition,parity,ambient temperatures,and farm management.Individual studies cannot cover all these factors.With the view to address this challenge,we searched all studies that were published from 1986 to 2020,and performed a systematic review and meta-analysis on the fat effect.In total,19 papers were collected and analyzed.Fat supplementation in sow diets during late gestation and lactation decreased average daily feed intake(ADFI,P<0.05)and tended to increase average daily energy intake(ADEI,P=0.11).It had no impact on litter weights at birth(P=0.40)or weaning(P=0.46).It increased total numbers of piglets at birth(P=0.07),but had no effect on liveborn per litter(P=0.90)or survival rate(P=0.48)of piglets to weaning.Fat supplementation had no significant effect on sow body weight loss(P=0.67)or backfat thickness changes(P=0.66),but sows fed diets with added fat had increased milk fat concentration(P=0.03)and shorter wean to estrus intervals(WEI,P=0.01).In specific circumstances,fat supplementation tended to improve growth performance of piglets with low litter weights at birth(P=0.14),or when the sows lost large amounts of body weight during lactation(P=0.11).The level of supplemented fat was 10%and higher would decrease liveborn per litter at neutral temperature(P=0.10).The meta-analysis revealed that fat supplementation to sows diet during late gestation and lactation can be beneficial for sow reproductive performance and litter growth performance.

Influences of dietary protein sources and crude protein levels on intracellular free amino acid profile in the longissimus dorsi muscle of finishing gilts

Background： The current study was carried out to determine effects of dietary protein source and crude protein（CP）level on carcass characteristics, meat quality, and muscle amino acid（AA） profile in finishing gilts. The experiment was designed as a 2 × 2 factorial arrangement with two sources of dietary proteins（cottonseed meal, CSM vs. soybean meal, SBM） and two levels of CP（12 % vs. 14 %, as-fed basis）. Seventy-two crossbred gilts（89.5 ± 0.9 kg） were allotted to one of four dietary treatments in a randomized complete block design for a period of 28 d. All diets were formulated to be isoenergetic and similar concentrations of standardized ileal digestible essential AA covering the nutrient requirements of pigs.Results： Growth, carcass characteristics and meat quality were not affected by dietary protein source nor crude protein level（P ＆gt; 0.10） except that average daily feed intake was increased by CSM diets（P = 0.03）. Gilts offered reduced protein diets had lower muscle p H45min（P 〈 0.05）. Neither dietary protein source nor crude protein level influenced N deposition. However, reduced protein diets decreased N intake, N excretion, and serum urea nitrogen content, whilst improved N efficiency（P 〈 0.01）. CSM diets increased N intake（P = 0.04）,but did not depress N efficiency. The concentrations of phenylalanine, tryptophan, cysteine and tyrosine（P 〈 0.05） of the longissimus muscle were decreased when gilts offered CSM diets, while muscle intracellular free valine concentration was increased（P = 0.03）. The gilts offered reduced protein diets had greater intracellular concentrations of free methionine, lysine, and total AA in muscle（P 〈 0.05）.Conclusion： These results suggest that CSM could replace SBM as a primary protein source in finishing pig diets in terms of performance, N efficiency, carcass characteristics, and meat quality, but decrease the concentrations of muscle specific AA. Furthermore, the reduced protein diet played an important role in increasing muscle intracellular concentrations of specific free amino acids（FAA）, and in reducing the relative ratios of specific FAA to lysine in longissimus dorsi muscle of pig, whose biological meaning needs further studies.

Ryanodine receptor RyR1-mediated elevation of Ca^(2+)concentration is required for the late stage of myogenic differentiation and fusion

Background:Cytosolic Ca^(2+)plays vital roles in myogenesis and muscle development.As a major Ca^(2+)release channel of endoplasmic reticulum(ER),ryanodine receptor 1(RyR1)key mutations are main causes of severe congenital myopathies.The role of RyR1 in myogenic differentiation has attracted intense research interest but remains unclear.Results:In the present study,both RyR1-knockdown myoblasts and CRISPR/Cas9-based RyR1-knockout myoblasts were employed to explore the role of RyR1 in myogenic differentiation,myotube formation as well as the potential mechanism of RyR1-related myopathies.We observed that RyR1 expression was dramatically increased during the late stage of myogenic differentiation,accompanied by significantly elevated cytoplasmic Ca^(2+)concentration.Inhibition of RyR1 by siRNA-mediated knockdown or chemical inhibitor,dantrolene,significantly reduced cytosolic Ca^(2+)and blocked multinucleated myotube formation.The elevation of cytoplasmic Ca^(2+)concentration can effectively relieve myogenic differentiation stagnation by RyR1 inhibition,demonstrating that RyR1 modulates myogenic differentiation via regulation of Ca^(2+)release channel.However,RyR1-knockout-induced Ca^(2+)leakage led to the severe ER stress and excessive unfolded protein response,and drove myoblasts into apoptosis.Conclusions:Therefore,we concluded that Ca^(2+)release mediated by dramatic increase in RyR1 expression is required for the late stage of myogenic differentiation and fusion.This study contributes to a novel understanding of the role of RyR1 in myogenic differentiation and related congenital myopathies,and provides a potential target for regulation of muscle characteristics and meat quality.

Effects of the Ratio of Lysine to Digestible Energy Level in the Diet on the Expression and Activity of Transcription Factors Involved in Lipogenesis in Rongchang Pigs

This study was conducted to determine the effects of varying the ratio of lysine to digestible energy level On the activity and gene expression of the transcription factors peroxisome proliferator-activated receptor-γ （PPAR-γ） and CCAAT/enhancer-binding protein-or and -β （C/EBP-α and C/EBP-β） to better understand the regulatory mechanisms controlling adipogenesis in fat and muscle tissue of the Rongchang pig. A total of 144 castrated Rongchang pigs weighing approximately 20 kg were used in a 2 ×2 factorial design experiment. Diets were formulated to contain a high （14.22 MJ/kg） or low （13.11 MJ/kg） digesti- ble energy （DE） level. Within each energy level, pigs were fed diets containing a high lysine： DE ratio （0.67,0. 53, or 0. 42） or a low lysine ： DE ratio （0.49,0.38 ,or 0.30） during the periods from 20 to 50 kg, 50 to 80 kg, and 80 kg to slaughter, respectively. Each diet was fed to six replicate pens, each containing nine pigs. When the pigs reached average live weights of 20,35,60, and 90 kg ,one pig from each of the replicates was chosen at random and slaughtered.Samples of back fat and longissimus dorsi muscle were collected for the assessment of transcriptional factor. The results showed that feeding a high DE level significantly increased （ P 〈 0.05 ） the expression of PPAR-T at 60 and 90 kg in muscle and at 35,60, and 90 kg in back fat. Energy level also significantly increased the expression of C/EBP-fl at 35 and 60 kg in both muscle and back fat （ P 〈 0.05 ）. Higher dieta- ry lysine increased the expression of C/EBP-fl in muscle at 35 and 90 kg （ P 〈 0.05）, but decreased the expression in back fat at 35 （P = 0.03 ） and 90 kg （P = 0.09）. The lysine level increased the expression of PPAR-3~ in muscle at 60 kg only. Energy level and lysine content had no significant effects on promote the activity of PPAR-γ, C/EBP-α, or C/EBP-β either in muscle or in back fat at any level of the body weights tested. Collectively, these data indicated that dietary energy density and lysine level were equally important for lipid deposition in muscle tissue, whereas dietary energy density was more important than lysine level for fat deposition in fat tissue.

Effect of dietary stable isotopic ratios of carbon and nitrogen on the extent of their incorporation into tissues of rats

This study was conducted to investigate the effect of different dietary ratios of 13C to 12C or 15 N to 14N on their relative incorporation into tissues. Eighty male rats were used in two 21-day feeding trials in which they were fed diets with either high δ13C levels （δ13C=-13.89%o and δ15N =2.37%o in experiment 1 and δ13C=-19.34%o and δ15N = 4.73%o in experiment 2） or low 613C levels （δ13C =-17.90%o and δ15N = 3.08%o in experiment 1 and δ13C =-21.76%o and δ15N = 0.53‰ in experiment 2）, meanwhile, the dietary δ15N levels were designed to two ranks Blood, liver, adipose and muscle tissues were collected on day 0, 3, 7, 14, and 21 for determination of 13C, 12C, 15N and 14 N isotopes. Rat growth rate, antioxidant capacity and metabolic parameters were also assessed. The results indicate that adipose tissue tend to deplete 13C before the stable isotopic ratios achieved final equilibrium. Therefore, feeds with different isotopic signatures had different incorporation rates into tissues. Low dietary 13C levels decreased tissue δ13C values whereas high dietary 13C levels did not alter tissue δ13C values during the 21-d experiment. Blood δ15N values were a reliable parameter in assessing the relative contribution of dietary nitrogen to tissues. This study revealed a relationship between dietary tissues. However, more studies are needed to illustrate the sotopic signatures and their incorporation rates into rat mechanism through which dietary isotopic ratios nfiuence the extent of isotopic incorporation into the tissues

Nutritional regulation of skeletal muscle energy metabolism, lipid accumulation and meat quality in pigs

The quality of pork determines consumers’ purchase intention, which directly affects the economic value of pork. Minimizing the proportion of inferior pork and producing high quality pork are the ultimate goals of the pig industry. Muscle energy metabolism, serving as a regulative hub in organism energy expenditure and storage as a fat deposit, is compatible with myofiber type composition, affecting meat color, intramuscular fat content, tenderness, p H values and drip loss. Increasing data illustrate that dietary nutrients and bioactive ingredients affect muscle energy metabolism, white adipose browning and fat distribution, and myofiber type composition in humans, and rodents. Recently, some studies have shown that modulating muscle energy metabolism and lipid accumulation through nutritional approaches could effectively improve meat quality. This article reviews the progress and development in this field, and specifically discusses the impacts of dietary supply of amino acids, lipids, and gut microbiota as well as maternal nutrition on skeletal muscle energy metabolism, lipid accumulation and meat quality of pigs, so as to provide comprehensive overview with respect to effective avenues for improving meat quality.

Effects of daidzein on antioxidant capacity in weaned pigs and IPEC-J2cells

Our previous study found that soybean isoflavones in soybean meal play an important role in improving growth performance and antioxidant capacity in pigs.However,it is still unknown whether long-term supplementation with daidzein,an active molecule deglycosylated from daidzin,in a corn-soybean meal diet can enhance growth performance in pigs.Thus,in the present study,an animal trial was carried out to investigate the effects of dietary supplementation with daidzein on the growth performance and antioxidant capacity of pigs.A total of 80 weaned piglets(40 barrows and 40 females)were assigned to 4treatments with 5 pens per treatment and 4 piglets per pen and fed a diet supplemented with 0,25,50 and 100 mg/kg daidzein for a 72-day trial.In addition,porcine intestinal epithelial cells(IPEC-J2)were used as an in vitro model to explore the underlying antioxidant mechanisms of daidzein.IPEC-J2 cells were treated with 0.6 mM hydrogen peroxide(H_(2)O_(2))in the presence or absence of 40μM daidzein.The results showed that adding 50 mg/kg of daidzein to the diet significantly improved body weight on day72,average daily gain(ADG)during days 0 to 72 and plasma superoxide dismutase(SOD)activity on day42,and decreased malondialdehyde(MDA)content on day 14(P<0.05).Treatment with 0.6 m M H_(2)O_(2)for 1 h significantly decreased cell viability and catalase(CAT)activity and increased intracellular reactive oxygen species(ROS)levels and MDA content(P<0.05),while pretreatment with 40μM daidzein prevented the decrease in cell viability and CAT activity and the increase in intracellular ROS levels and MDA content caused by H_(2)O_(2)(P<0.05).In addition,H_(2)O_(2)stimulation significantly suppressed the expression of nuclear factor erythroid-2-related factor 2(Nrf2),CAT,occludin and zonula occludens-1(ZO-1),while pretreatment with daidzein preserved the expression of Nrf2,CAT and occludin in H_(2)O_(2)-stimulated IPEC-J2 cells(P<0.05).In conclusion,our results suggested that long-term dietary supplementation with 50 mg/kg daidzein improved growth performance in pigs and was beneficial to the antioxidant capacity of pigs.Daidzein exerted protective effects against H_(2)O_(2)-induced oxidative stress in IPEC-J2 cells and the underlying mechanism may be related to the activation of the Nrf2 signaling pathway.

Arginine promotes myogenic differentiation and myotube formation through the elevation of cytoplasmic calcium concentration

This study aimed to explore the mechanism underlying arginine-promoted myogenesis of myoblasts.C2C12 cells were cultured with a medium containing 0.1,0.4,0.8,or 1.2 mmol/L arginine,respectively.Cell proliferation,viability,differentiation indexes,cytoplasmic Ca^(2+)concentration,and relative mRNA expression levels of myogenic regulatory factors(MRF)and key Ca2+channels were measured in the absence or presence of 2 chemical inhibitors,dantrolene(DAN,10μmol/L)and nisoldipine(NIS,10μmol/L),respectively.Results demonstrated that arginine promoted myogenic differentiation and myotube formation.Compared with the control(0.4 mmol/L arginine),1.2 mmol/L arginine upregulated the relative mRNA expression levels of myogenin(MyoG)and Myomaker at d 2 during myogenic induction(P<0.05).Cytoplasmic Ca^(2+)concentrations were significantly elevated by arginine supplementation at d 2 and 4(P<0.05).Relative mRNA expression levels of Ca^(2+)channels including the type 1 ryanodine recepto r(RyR1)and voltage-gated Ca^(2+)channel(Cav1.1)were upregulated by 1.2 mmol/L arginine during2-d myogenic induction(P<0.01).However,arginine-promoted myogenic potential of myoblasts was remarkably compromised by DAN and NIS,respectively(P<0.05).These findings evidenced that the supplementation of arginine promoted myogenic differentiation and myotube formation through increasing cytoplasmic Ca^(2+)concentration from both extracellular and sarcoplasmic reticulum Ca^(2+).

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(T23peak 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.