Plasma free amino acid profiles of canine mammary gland tumors

The purpose of this study was to elucidate the relationship between plasma free amino acid (PFAA) levels and the clinical stages of mammary gland tumors (MGT) in dogs. PFAA levels in canines with malignant mammary tumors were decreased compared to those of healthy animals. The levels of aspartate and ornithine, in the dogs with tumor metastasis were significantly decreased when compared to those of dogs that did not have metastases. Results of this study indicate that PFAA levels could be a risk factor or biomarker for canine MGT metastasis.     

Branched‐chain amino acid ratios in low‐protein diets regulate the free amino acid profile and the expression of hepatic fatty acid metabolism‐related genes in growing pigs

Liver metabolism is affected by nutrients. The aim of this study was to explore the effects of low‐protein diets (17% crude protein, CP) supplemented with branched‐chain amino acids (BCAAs), including leucine (Leu), isoleucine (Ile) and valine (Val), on hepatic amino acid profile and lipid metabolism in growing pigs. The ratio of Leu : Ile : Val in all groups was 1 : 0.51 : 0.63 (20% crude protein, CP), 1 : 1 : 1 (17% CP), 1 : 0.75 : 0.75 (17% CP), 1 : 0.51 : 0.63 (17% CP) and 1 : 0.25 : 0.25 (17% CP) respectively. Results revealed that compared to the positive control group (1 : 0.51 : 0.63, 20% CP), the low‐protein diets significantly augmented the concentrations of most essential amino acids and non‐essential amino acids (p < .05), with the greatest values observed in the 1 : 0.25 : 0.25 group. Moreover, relative to the control, the low‐protein diets with the Leu : Ile : Val ratio ranging from 1 : 0.75 : 0.75 to 1 : 0.25 : 0.25 markedly downregulated the mRNA abundance of acetyl‐CoA carboxylase (ACC), lipoprotein lipase (LPL) and fatty acid‐binding protein 4 (FABP‐4) (p < .05), and upregulated the mRNA expression of hormone‐sensitive lipase (HSL), peroxisome proliferator‐activated receptor‐g coactivator‐1α (PGC‐1α), uncoupling protein 3 (UCP3) and liver carnitine palmitoyltransferase 1 (L‐CPT‐1) (p < .05). Therefore, our data suggest that protein‐restricted diets supplemented with optimal BCAA ratio, that is, 1 : 0.75 : 0.75–1 : 0.25 : 0.25, induce a shift from fatty acid synthesis to fatty acid oxidation in the liver of growing pigs. These effects may be associated with increased mitochondrial biogenesis.     

Effect of excess levels of lysine and leucine in wheat‐based,amino acid‐fortified diets on the mRNA expression of two selected cationic amino acid transporters in pigs

An experiment was conducted to evaluate the effect of excess levels of Leu and Lys on the expression of b^0,+ and CAT‐1 mRNA in jejunum, liver and the muscles Longissimus dorsi (LDM) and Semitendinosus (STM). Twenty pigs with an average initial BW of 16.4 ± 1.7 kg were used in a Randomized Complete Block. Dietary treatments (T) were as follows: T1, basal diet; T2, basal plus 3.5 g l ‐Lys/kg diet; T3, basal plus 1.5 g l ‐Leu/kg diet; T4, basal plus 3.5 g l ‐Lys plus 1.5 g l ‐Leu/kg diet. Diets in T1 and T3 met 100% the requirement of Lys for pigs within the 10 to 20 kg body weight range; diets in T2 and T4 contained 35% excess of Lys. Also, diets in T1 and T2 supplied 104%, whereas diets in T3 and T4 supplied 116% the requirement of Leu. The expression of b^0,+ in jejunum was reduced (p = 0.002) because of the supplementation of l ‐Leu, but l ‐Lys supplementation had no effect (p = 0.738). In contrast, the expression of b^0,+ in STM (p = 0.012) and liver (p = 0.095) was reduced by the high level of Lys, but Leu had no effect (p > 0.100). CAT‐1 expression in STM increased by high Lys (p = 0.023) and Leu (p = 0.007) levels. In liver, the expression of CAT‐1 substantially increased (p = 0.001) because of Lys. In conclusion, excess levels of dietary Lys and Leu affect the expression of cationic amino acid transporters, and this effect varies depending on the studied tissue.     

L‐carnitine improves hydrogen peroxide‐induced impairment of nuclear maturation in porcine oocytes

We investigated the effect of oxidative stress induced by hydrogen peroxide (H₂O₂) on lipid peroxide (LPO) level and nuclear maturation in porcine oocytes cultured with or without cumulus cells. After 22 h of pre‐culture, oocytes with attached cumulus cells (COC group) or denuded oocytes (DO group) were cultured with H₂O₂, and intra‐oocyte H₂O₂ and LPO levels were quantitatively analyzed using immunofluorescence. This is the first report evaluating LPO levels in porcine oocytes. After H₂O₂ supplementation, the DO group showed severe accumulation of H₂O₂ and LPO in the oocytes. Similarly, while inhibition of progression of nuclear maturation was observed in both groups, the effect was more severe in the DO group. These results demonstrate that cumulus cells reduce the accumulation of H₂O₂ stress in oocytes. Furthermore, we attempted to reduce the oxidative stress by H₂O₂ with L‐carnitine, a H₂O₂ scavenger. L‐carnitine decreased H₂O₂ and LPO levels in the oocytes in both groups, and improvement in the progression of impaired nuclear maturation was observed. These effects were different by the presence of cumulus cells. Our results provide that L‐carnitine is useful for alleviating H₂O₂‐induced oxidative stress by reducing LPO levels and improving the progression of nuclear maturation.     

Effect of L‐theanine on meat quality,muscle amino acid profiles,and antioxidant status of broilers

This study investigated the effect of L ‐theanine on carcass traits, meat quality, muscle antioxidant capacity, and amino acid (AA) profiles of broilers. Three hundred 1‐day‐old Ross 308 male broilers were randomly allotted to five groups with six replicates. Birds were fed the basal diet or basal diet with 300, 600, 900, or 1,500 mg/kg L ‐theanine for 42 consecutive days. The results showed that L ‐theanine quadratically increased dressing percentage, eviscerated percentage, and leg muscle yield (p < .05). Meanwhile, drip loss, cooking loss, shear force, L*_24h, and muscle lactate content decreased quadratically in response to dietary L ‐theanine supplementation (p < .05), while pH_24h and muscle glycogen content were quadratically improved by L ‐theanine (p < .05). Notably, the contents of muscle malondialdehyde and protein carbonyl, and the activities of muscle total antioxidant capacity, catalase, and glutathione peroxidase decreased quadratically in response to dietary L ‐theanine supplementation (p < .05), suggesting that the oxidative stress level of muscle was decreased quadratically. Moreover, L ‐theanine quadratically increased the concentrations of most of muscle essential AA, nonessential AA, and flavor AA (p < .05). In conclusion, L ‐theanine can be used as a valuable feed additive to modulate carcass traits, meat quality, muscle antioxidant status, and AA profiles of boilers, and its optimum addition level is 600 mg/kg based on the present study.     

Supplementation of L‐glycine and L‐glutamate to Japanese quails from 01 to 36 days of age using the ideal protein concept

Experiment I: T_1‐1 = basal diet with 25% crude protein (CP) + limiting amino acids (LA); T_1‐2 = 20% CP + LA; T_1‐3 = 20% CP + LA + L‐glycine; T_1‐4 = 20% CP + LA + L‐glutamate; T_1‐5 = 20% CP + LA + L‐glycine + L‐glutamate. Experiment II: T_2‐1 = basal diet with 22% CP + LA; T_2‐2 = 20% CP + LA; T_2‐3 = 17.6% CP + LA + L‐glycine; T_2‐4 = 17.6% CP + LA + L‐glutamate; T_2‐5 = 17.6% CP + LA + L‐glycine + L‐glutamate. The reduction of dietary protein based on the concept of ideal protein decreases nitrogen excretion in quails when L‐glycine is added to the diets. Quails fed diets supplemented with L‐glutamate as the non‐specific nitrogen source equivalent to the nitrogen level of the control diet had increased nitrogen excretion. However, quails had reduced nitrogen excretion in both experiments when L‐glycine was added to diets with L‐glutamate. Carcass fat was increased by reducing dietary protein, but fat deposition was reduced by adding L‐glutamate and L‐glycine, or both. The dietary addition of L‐glutamate and L‐glycine in quails based on the ideal protein concept is not necessary (Exp. I). Although the total nitrogen, electrolytic balance and glycine level were adjusted in diets, quails had decreased performance. Therefore, other hypotheses besides protein reduction need to be studied (Exp. II). Protein reduction with supplementation of only limiting essential amino acids does not affect quail performance. Dietary addition of L‐glycine reduces nitrogen excretion.     

Association of the amino acid motifs of BoLA-DRB3 alleles with mastitis pathogens in Japanese Holstein cows

The association of the polymorphism of bovine leukocyte antigen ( BoLA-DRB3 ) genes, identified by the polymerase chain reaction sequence-based typing (PCR-SBT) method, with resistance and susceptibility to mastitis caused by Streptococci , coagulase-negative Staphylococci, Escherichia coli and Staphylococcus aureus was investigated. Blood samples for DNA extraction were collected from 170 Holstein cows (129 mastitis and 41 healthy cows) from 5 districts in Chiba prefecture, Japan. Susceptibility or resistance to the mastitis-causing pathogens was thought to vary by the presence of amino acid substitutions at the 9, 11, 13, and 30 positions. DRB3*0101 and DRB3*1501 had amino acid motifs of Glu⁹, Ser¹¹, Ser¹³, and Tyr³⁰, and they were considered to have susceptibility to all 4 mastitis pathogens. In contrast, DRB3*1101 and DRB3*1401 had amino acid motifs of Gln⁹, His¹¹, Gly¹³, and His³⁰ in these positions, and they also had Val⁸⁶, so these alleles were considered to have resistance to Streptococcal and coagulase-negative Staphylococcal mastitis. However, in the case of Escherichia coli mastitis, amino acid substitutions at the 9, 11, 13, and 30 positions had little effect, but rather substitutions at the 47, 67 positions of pocket 7, and at the 71, 74 positions of pocket 4, Tyr⁴⁷, Ile⁶⁷, Ala⁷¹, and Ala⁷⁴, were associated with resistance. This motif was present in DRB3*1201 .     

Reduced dietary protein level influences the free amino acid and gene expression profiles of selected amino acid transceptors in skeletal muscle of growing pigs

This study was conducted to evaluate the effect of reduced dietary protein level on growth performance, muscle mass weight, free amino acids (FAA) and gene expression profile of selected amino acid transceptors in different fibre type of skeletal muscle tissues (longissimus dorsi, psoas major, biceps femoris) of growing pigs. A total of 18 cross‐bred growing pigs (Large White × Landrace × Duroc) with initial body weight (9.57 ± 0.67 kg) were assigned into three dietary treatments: 20% crude protein (CP) diet (normal recommended, NP), 17% CP diet (low protein, LP) and 14% CP diet (very low protein, VLP). The results indicated improved feed‐to‐gain ratio was obtained for pigs fed LP and NP diets (p < 0.01), while the pigs fed VLP diet showed the worst growth performance (p < 0.01). There was no significant difference in the weights of longissimus dorsi and psoas major muscle between LP and NP groups (p > 0.05). Majority of the determined FAA concentration of LP group were greater than or equal to those of NP group in both longissimus dorsi and psoas major muscle (p < 0.01). Further, the mRNA expression levels of sodium‐coupled neutral amino acid transceptor 2, L‐type amino acid transceptor 1 and proton‐assisted amino acid transceptors 2 were higher in skeletal muscle tissue in LP group compared to those of the pigs fed NP or VLP diet. These results suggested that reduced dietary protein level (3 points of percentage less than recommended level) would upregulate the mRNA expression of amino acid transceptors to enhance the absorption of FAA in skeletal muscle of growing pigs. There seems to be a relationship between response of AA transceptors to the dietary protein level in skeletal muscle tissue of different fibre type. To illustrate the underlying mechanisms will be beneficial to animal nutrition.     

Effect of L‐lysine on expression of selected genes,serum concentration of amino acids,muscle growth and performance of growing pigs

Lysine (Lys) is the first limiting amino acid (AA) in most feed formulations for pigs and most abundant, along with leucine, in muscle proteins. An experiment was conducted with 17 pigs (17.7 ± 0.05 kg initial BW) to identify a role of dietary Lys in the control of protein synthesis in pigs. Fourteen pigs were randomly assigned to one of the two wheat‐based dietary treatments: Lys‐deficient, 3.0 g/kg (DEF) and Lys‐adequate, 10.8 g/kg (ADE). Samples from jejunum mucosa, liver, Longissumus and Semitendinosus muscles, and blood were collected. The other three pigs were sacrificed at the beginning of the trial to measure basal carcass composition. Weight gain, gain:feed ratio, Lys intake and loin eye area were greater in ADE than in DEF pigs (p < 0.01). Muscle‐related carcass characteristics were better, and myosin heavy chain IIb expression (MyHC IIb) in Semitendinosus was higher in ADE than in DEF pigs. Expression of AA transporters CAT‐1 was lower (p < 0.05), serum Lys was higher and serum Val was lower in pigs fed the ADE diet. The higher muscularity, MyHC IIb expression in Semitendinosus muscle and Lys serum of pigs fed the ADE diet suggest that Lys increases growth rate not only by functioning as protein construction unit but also as potential control of the protein synthesis process.     

Immunolocalization of lingual antimicrobial peptide (LAP) in the bovine mammary gland

Lingual antimicrobial peptide (LAP), a member of the β-defensin family in cows, is involved in the innate immune system and plays a crucial role in killing a large variety of microorganisms. The aim of the present study was to demonstrate the immunolocalization of LAP in the mammary glands of cows. A LAP antibody was raised in a rabbit by immunity with a synthetic 11 amino acid sequence out of a 42-amino acid sequence of the mature form of LAP. The specificity of the LAP antibody was checked using a competitive immunoassay and Western blotting. Paraffin sections of the mammary gland were immunostained with LAP antibody. In the competitive immunoassay, an increase of synthetic LAP concentration suppressed the optical density. Western blotting analysis for LAP revealed the presence of the LAP peptide in mammary alveolar tissue. When the mammary gland was immunostained with LAP antibody, epithelial cells of both infected and non-infected alveoli were immunopositive. These results indicate that LAP is localized in the epithelium of non-infected as well as infected alveolus in the mammary gland in cows.     

Possibility of amino acid nutrition in broiler

The consumption of poultry meats exceeds 50% of total meat consumption. With the outbreak of bovine spongiform encephalopathy and foot and mouth disease, the consumption of poultry meat is set to increase. The use of meat bone meal derived from cattle and pigs is prohibited. Therefore, plant protein has to be used more frequently and the importance of the study of amino acid nutrition is increasing. In order to improve dietary efficiency, it is necessary to elucidate the exact amino acid requirements of poultry. The amino acid requirements are not constant and are affected by various factors. By using the plasma amino acid concentration as a parameter, it is possible to predict the amino acid requirements within a short period using a small number of animals repeatedly. By satisfying the amino acid requirements through supplementing crystalline amino acids, it is possible to reduce dietary protein and excretory nitrogen simultaneously. However, when the dietary protein is reduced, the abdominal fat tends to increase. This is solved by deciding the optimum dietary protein level, but it is difficult to control the taste of poultry meat by dietary amino acids.     

Dietary levels of protein and free amino acids affect pancreatic proteases activities,amino acids transporters expression and serum amino acid concentrations in starter pigs

The dietary contents of crude protein and free amino acids (AA) may affect the protein digestion and AA absorption in pigs. Trypsin and chymotrypsin activities, AA serum concentrations and expression of AA transporters in the small intestine of pigs fed a low protein, AA‐supplemented (19.2%, LPAA) or a high protein (28.1%, HP), wheat‐soybean meal diet were measured in two 14‐d trials. The LPAA diet contained free L‐Lys, L‐Thr, DL‐Met, L‐Leu, L‐Ile, L‐Val, L‐His, L‐Trp and L‐Phe. All pigs were fed the same amount of feed (890 and 800 g/d for trial 1 and 2 respectively). In trial 1, samples of mucosa (duodenum, jejunum and ileum) and digesta (duodenum and jejunum) were collected from 14 pigs (17.2 ± 0.4 kg); in trial 2, blood samples were collected from 12 pigs (12.7 ± 0.3 kg). The trypsin and chymotrypsin activities in both intestinal segments were higher in pigs fed the HP diet (p < 0.01). Trypsin activity was higher in jejunum than in duodenum regardless the dietary treatment (p < 0.05). Pigs fed the LPAA diet expressed more b^0,+AT in duodenum, B⁰AT1 in ileum (p < 0.05), and tended to express more y⁺LAT1 in duodenum (p = 0.10). In pigs fed the LPAA diet, the expression of b^0,+AT was higher in duodenum than in jejunum and ileum (p < 0.01), but no difference was observed in pigs fed the HP diet. Ileum had the lowest b^0,+AT expression regardless the diet. The serum concentrations of Lys, Thr and Met were higher in LPAA pigs while serum Arg was higher in HP pigs (p < 0.05). Serum concentrations of AA appear to reflect the AA absorption. In conclusion, these data indicate that the dietary protein contents affect the extent of protein digestion and that supplemental free AA may influence the intestinal site of AA release and absorption, which may impact their availability for growth of young pigs.     

Effects of eccentric exercise on branched‐chain amino acid profiles in rat serum and skeletal muscle

Supplementation of branched‐chain amino acid (BCAA) is often used to attenuate exercise‐induced skeletal muscle damage and promote adaptation, but no definitive conclusion on the benefits of BCAA on muscle recovery after injurious exercise can be drawn. Exploration of the systematic BCAA alteration in muscular injury‐repair stage per se without any BCAA supplement should provide some useful information in favour of BCAA application in muscle regeneration after injury. One bout of 90‐min downhill‐running exercise was performed to cause rat skeletal muscle injury. After exercise, myofibrillar BCAA concentrations showed minor changes compared with exercise before, while serum concentrations of BCAA were lower after exercise. Especially, serum leucine, isoleucine and total BCAA concentrations 2 weeks post‐run were significantly lower than normal values of exercise before (p = 0.008, p = 0.041, p = 0.015). The data demonstrate that a single eccentric exercise can significantly decrease the serum BCAA concentrations, which mean high utilization of BCAA for myogenesis after injurious exercise.     

过瘤胃氨基酸在反刍动物饲料中的应用

本文综述了过瘤胃赖氨酸及蛋氨酸在反刍动物饲料中的应用,旨为过瘤胃氨基酸在反刍动物生产中的合理使用提供理论依据。     

Effects of branched-chain amino acids on immune status of young racing horses

High-intensity exercise and competition are associated with depressed immune function. Young horses, which participate in high-intensity exercise and competitions, are at increased risk for the development of infectious disease due to depression of immune function. The effects of branched-chain amino acid (BCAA) supplementation on the immune status of young racing horses were evaluated, determining whether BCAA might help to avoid or reduce immune suppression during exercise and competitions. Twenty horses (10 male and 10 female) were treated with BCAA supplementation; another twenty untreated horses (10 male and 10 female) constituted control group. Peripheral blood was collected from each animal and evaluated for lymphocyte subsets, phagocytosis analysis of monocytes and granulocytes, lymphocyte proliferative response, and expression of cytokine-encoding messenger ribonucleic acids (mRNAs). The numbers of CD4⁺, CD8⁺, and major histocompatibility complex (MHC) class II⁺ cells in females of the treated group were significantly higher than those in females of the control group. The lymphocyte proliferative response in female of the treated group also was significantly higher than that in females of the control group. In addition, expression of mRNAs encoding interleukin-1β (IL-1β) and interferon-γ (IFN-γ) in females of the treated group was significantly higher than that in females of the control group. There were no significant differences between males of the treated and control groups. The results of this study indicated the positive effects of BCAA supplementation in counteracting immunosuppression in young female racing horses during and following high-intensity exercise.     

盐碱胁迫下外源硫化氢对裸燕麦叶片氨基酸代谢过程的影响

信号硫化氢缓解植物盐碱胁迫机理备受关注。为探讨外源硫化氢对盐碱胁迫下植物氨基酸代谢的调控机制,采用盆栽土培试验,以裸燕麦品种‘定莜9号’为材料,设置盆土不添加盐碱和添加3.00 g·kg^-1盐碱(摩尔比NaCl︰Na₂SO₄︰Na₂CO₃︰Na HCO₃=12︰8︰1︰9)与抽穗期叶面喷蒸馏水和喷50μmol·L^-1硫化氢供体硫氢化钠(NaHS)溶液,共4个处理。研究其对叶片中总氨基酸、丙二醛含量和籽粒产量的影响;运用液相色谱和质谱检测,采用主成分分析22种组成蛋白质的氨基酸中差异氨基酸,解析外源硫化氢对氨基酸代谢途径的调控效应。结果表明：在裸燕麦叶片中未检出高半胱氨酸。喷施NaHS溶液对盐碱胁迫下裸燕麦叶片中总氨基酸含量下降的缓解效应不显著,对盐碱胁迫诱导的丙二醛含量的升高和籽粒产量的下降有显著的缓解作用。主成分分析结果显示：盐碱胁迫下,喷施NaHS可显著下调裸燕麦叶片中α-酮戊二酸族的鸟氨酸和草酰乙酸族的天冬酰胺含量;显著上调α-...     

Repeated restraint stress modifies fatty acid and amino acid metabolism in the mouse skin

In modern society, stress caused by relationships and emotions is one of the greatest social problems. Similar to humans, domestic and captive animals live under various stresses. Several stresses have been associated with skin disorders, such as atopic dermatitis, but there is a lack of reliable and objective indicators for the characterization of this association. This study aimed to define the changes in fatty acid composition and amino acid concentration in the skin following repeated restraint stress in ICR mice. Mice subjected to 30 min of daily restraint stress for 8 days showed changes in the composition of saturated fatty acids, such as an increase in palmitic acid content, which are the substrates of Δ-9 desaturase. Conversely, unsaturated fatty acids decreased with stress treatment, which appeared to be a result of these fatty acids being the substrate of Δ-6 desaturase. Changes in fatty acid composition after stress treatment may be one of the factors that cause skin inflammation. The water-retention capacity may have been lowered by stress treatment because histidine and leucine, which are natural moisturizing factors, were significantly decreased. The collagen content in the skin gradually decreased after repeated stress treatment. Our results indicate that repeated restraint stress may impact skin health through changes in both the fatty acid composition and amino acid concentration in mice.     

影响家禽可消化氨基酸体系应用的因素

本文就可消化氨基酸体系应用于家禽日粮中存在的技术问题和理论问题进行了分析,并对国内外的相关研究进行了综述。