Dietary β-alanine enhances brain, but not muscle, carnosine and anserine concentrations in broilers

Carnosine (β‐alanyl‐L‐histidine) and its derivative anserine (β‐alanyl‐1‐methyl‐L‐histidine) are present in high concentrations in the muscle and brain of chickens. They are known as antioxidants and putative neurotransmitters in the brain. If administration of β‐alanine (β‐Ala), one of the constituents of carnosine, could increase the concentrations of these dipeptides in the brain and muscles, it could improve brain function and increase the commercial value of the meat in chickens. As an early step in investigating this hypothesis, in the present study, the effect of dietary β‐Ala on these dipeptide concentrations in the brain, Musculus pectoralis superficialis, Musculus pectoralis profundus and Musculus biceps femoris was investigated in broilers. Four‐week‐old broilers were given a commercial diet or diet containing 0.5, 1 or 2%β‐Ala for 4 weeks. At the end of the experiment, concentrations of both dipeptides were increased in the brain, while taurine concentration was decreased. In the muscles, concentrations of these dipeptides were unchanged. These results indicate that dietary β‐Ala might influence brain function, but is ineffective in increasing the concentrations of carnosine and anserine in the muscles of broilers.     

Increased broiler muscle carnosine and anserine following histidine supplementation of commercial broiler feed concentrate

Abstract

A feeding experiment involving histidine supplementation to broiler feed resulted in increased concentration of the histidine containing dipeptides anserine and carnosine in broiler breast muscle. Supplementation with 1 g histidine per kg feed gave a 64% increase in carnosine, and about 10% increase in anserine in the muscle. The standard broiler feed concentrate now in use in Norway seems to contain less histidine than what may be needed for optimal synthesis of carnosine and anserine. These dipeptides have important roles as antioxidants, pH buffering agents and anti-glycation agents. They may have important roles in meat for increasing its stability, shelf life and antioxidant capacity, and it might be speculated that broiler meat rich in anserine and carnosine in the future will be considered a type of functional food, having possible health-beneficial effects. Histidine supplementation of standard Norwegian broiler feed concentrate should be considered.     

Antioxidation status and histidine dipeptides content in broiler blood and muscles depending on protein sources in feed

One‐day‐old chickens were fed mixtures containing different raw materials (fish by‐products meal, porcine blood cells meal, blood meal, wheat gluten, fodder yeast), as a source of histidine and β‐alanine – components of carnosine. Control birds were administered a feed mixture, in which soy bean meal was the main protein source. The bodyweight, feed consumption and conversion, antioxidant characteristics and histidine dipeptides content in blood and muscles, and also amino acid composition of chicken meat on day 34 post‐hatch were recorded. The best (p < 0.05) performance and feed conversion were observed in chickens fed mixture containing porcine blood cells meal. In blood plasma of control chickens, a significantly (p < 0.01) higher ability to scavenge DPPH radicals was found. However, the highest catalase activity in erythrocytes was determined in chickens fed mixtures with blood by‐products. Insignificant differences in both carnosine and anserine levels in plasma between treatments were noted. Breast muscles from control birds were characterized by lower activity of glutathione peroxidase (GPx), superoxide dismutase (SOD) and catalase (CAT) (p < 0.05; p < 0.01), than those from chickens fed blood by‐products. Improved ability to reduce ferric ions (FRAP) (p < 0.01) and carnosine content in meat from chickens fed blood cell meal were recorded. No direct relations between amino acids content in feed mixtures and in meat were observed.     

Concentrations of carnosine, anserine, L-histidine and 3-methyl histidine in boar spermatozoa and sheep milk by a modified HPLC method

The present study deals with the application of high-performance-liquid-chromatography (HPLC) method for a quantitative detection of carnosine, anserine, L-histidine and 3-methyl-L-histidine in biological material with o-phthaldialdehyde (OPA) post column derivatisation at the constant temperature of 50 degrees C. For this purpose, some mobile-phases were prepared with scalar acetonitrile concentrations. A complete separation of all molecules, particularly for carnosine and 3-methyl-L-histidine, was obtained with a solution of acetonitrile and 6mM hydrochloric acid with 0.48 M sodium chloride (5%:95% v/v). Post column derivatisation reaction at temperature of 50'C permitted to obtain an increase in sensibility of all molecules. This method has been utilised for detection of histidine dipeptides in boar spermatozoa and in sheep milk. Concentrations (mean +/- S.E. nmol/10(9) spermatozoa) of carnosine (0.96 +/- 0.14) and anserine (0.83 +/- 0.18) in boar spermatozoa were significantly lower than those of L-histidine (52.85 +/- 4.86) and 3-methyl-L-histidine (83.07 +/- 7.1). Positive correlation was found between carnosine and anserine contents (r = 0.740; p < 0.01) and between L-histidine and 3-methyl-L-histidine (r = 0.657; p < 0.01). All histidine dipeptides studied were also present in 40 samples of sheep milk. In a case of samples without unit-forming colonies (UFC) of Staphylococcus coagulase-positive, carnosine concentrations (9.17 +/- 0.89 nmol/ml) were higher than anserine (0.51 +/- 0.02 nmol/ml) and both were significantly lower in respect to L-histidine (49.51 +/- 6.48 nmol/ml) and 3-metyl-L-histidine (81.21 +/- 6.82 nmol/ml). A negative correlation was observed between carnosine milk levels (r = -0.773; p < 0.01) and UFC/ml of Staphylococcus coagulase-positive. In conclusion this very simple and fast method can be used to detect histidine dipeptides in biological compartments where their concentrations are very low.     

Oral administration of β-alanine modifies carnosine concentrations in the muscles and brains of chickens

Carnosine, and its derivative anserine, are known to function as anti‐oxidants and putative neurotransmitters. They are especially rich in the breast muscle (Musculus pectoralis superficialis, MPS) of chickens. To clarify whether the concentrations of carnosine and anserine are altered by dietary management, the effect of oral administration of their constituent, β‐alanine (β‐Ala), was determined in the MPS and brains of chickens. Birds were orally administered β‐Ala (22 mmol/kg) twice a day for five consecutive days (from 2 to 6 days old). In the MPS, carnosine was increased by β‐Ala, whereas anserine and taurine were decreased. The concentrations of other free amino acids in the MPS were also modified by β‐Ala. In the brain, the oral administration of β‐Ala increased anserine and carnosine and decreased taurine, but caused no changes to other free amino acid concentrations. These results suggest that orally administered β‐Ala increases carnosine concentrations in both the MPS and brains of chickens. However, the effects of β‐Ala on other concentrations differ depending on the tissues.     

Dietary supplementation with carnosine improves antioxidant capacity and meat quality of finishing pigs

This study was conducted to determine the effect of dietary carnosine (β‐alanyl‐l ‐histidine) supplementation on antioxidant capacity and meat quality of pigs. 72 pigs approximately 60 kg were fed a corn‐ and soybean meal‐based diet supplemented with 0, 25, 50 or 100 mg carnosine per kg diet for 8 weeks. Carnosine supplementation did not affect growth performance and carcass traits of pigs. However, the addition of 100 mg carnosine per kg diet increased pH value of muscle at 45 min, 24 h and 48 h postmortem. It also decreased drip loss at 48 h postmortem and increased redness value of muscle at 45 min postmortem (p < 0.05). The addition of 100 mg carnosine per kg diet enhanced glycogen concentration and Ca‐ATPase activity at 24 and 48 h postmortem, and reduced malondialdehyde and carbonyl protein complexes concentrations in muscle at 24 h postmortem (p < 0.05). The addition of 100 mg carnosine per kg diet increased glutathione peroxidase (GSH‐Px), superoxide dismutase (SOD) and catalase (CAT) activities in plasma, liver or muscle, as well as SOD and GSH‐Px genes expression in muscle (p < 0.05). Taken together, these findings indicate that carnosine supplementation improves antioxidant capacity and meat quality of pigs.     

Effect of Anserine and Carnosine on Sperm Motility in the Japanese Quail

Sperm motility is considered as one of the most important traits for successful fertilization, but the motility of an ejaculated sperm decreases with time when stored as liquid. It is reported that seminal plasma serves as a nutrient rich medium for sperm and plays an important role in sperm motility and its fertilization ability. Several studies have reported that imidazole dipeptides such as anserine and carnosine affect sperm motility and its fertilization ability in mammals. In this study, we report the presence of anserine and carnosine in the male reproductive tract of the Japanese quail. Abundant levels of anserine (44.46 µM) and carnosine (41.75 µM) were detected in the testicular fluid and seminal plasma respectively using the amino acid analyzer; however, seminal plasma solely contained carnosine. When the ejaculates were incubated with anserine or carnosine, we found that both the dipeptides improve sperm motility parameters such as straight line velocity, curvilinear velocity, average path velocity and amplitude of lateral head displacement after in vitro sperm storage at 15°C. These results indicate that imidazole dipeptides are present in the male reproductive tract and may improve sperm quality during in vitro sperm storage in the liquid states.     

Effects of slaughter age on the levels of free amino acids and dipeptides in fattening cattle

The effects of slaughter age, breed type and postmortem period were studied on the concentrations of free amino acids (FAA) and dipeptides (carnosine and anserine) in Longissimus dorsi muscle of beef. Slaughter age affected the levels of most FAA and dipeptides, and the concentrations of these compounds were significantly lower at 35 months of age than either 15 or 25 months of age. Slight increases were observed at 25 months compared with 15 months, but the differences were not significant except for a few FAA. These tendencies still remained when the concentrations were recalculated on the basis of protein. A significant breed effect was observed for taurine, carnosine and anserine. Almost all FAA were significantly increased during postmortem conditioning, but the level of each FAA fluctuated, and the fluctuations could not be explained statistically by slaughter age or breed differences.     

Use of histidine dipeptides and myoglobin to monitor adulteration of cooked beef with meat from other species

A new high performance liquid chromatography (HPLC) method was used to monitor the adulteration of cooked beef products with meat from other species. The ratio of the histidine dipeptides anserine and carnosine which are present in skeletal muscle, are so different between sheep, cattle, horse and kangaroo that detection of adulteration can be rapidly achieved by chromatography on a Partisil-10 SCX column with 0.2 M lithium formate, pH 2.9. To obtain a definitive identification of the adulterant it was necessary to also examine the electrophoretic mobility of myoglobin in sodium dodecylsulphate gels. One brand of "beefsteak" pie was found to actually be a mixture of mutton and beef.     

L-组氨酸对肉鸡生长性能、肌肉品质、血浆抗氧化能力和肌肉咪唑二肽含量的影响

本试验旨在研究饲粮添加L-组氨酸对肉鸡生长性能、肌肉品质、血浆抗氧化能力和肌肉咪唑二肽含量的影响。选取体重相近、健康的1日龄爱拔益加(AA)肉仔鸡公雏300只,随机分为5组,每组6个重复,每个重复10只鸡。对照组饲喂基础饲粮,试验组饲喂在基础饲粮中分别添加250、500、1 000、2 000 mg/kg L-组氨酸的试验饲粮。试验期42 d。结果表明:1)L-组氨酸降低了肉鸡生长后期(22~42日龄)的料重比,250和2 000 mg/kg添加组显著低于对照组(P0.05)。2)L-组氨酸对肌肉全净膛率、胸肌率、腿肌率、腹脂率、滴水损失、剪切力和pH无显著影响(P0.05)。3)L-组氨酸线性提高了42日龄血浆总抗氧化能力及谷胱甘肽过氧化物酶、总超氧化物歧化酶的活性(P0.05)。4)L-组氨酸增加了42日龄肌肉中肌肽和鹅肌肽含量,500、1 000和2 000 mg/kg添加组肌肽含量分别较对照组升高了40.58%、42.75%、33.70%(P0.05),1 000 mg/kg添加组鹅肌肽含量较对照组增加了37.80%(P0.05);对肌肉中肌肽和鹅肌肽含量进行二次曲线方程拟合表明,L-组氨酸添加量为1 276 mg/kg时,肌肉咪唑二肽含量最高。可见,饲粮添加L-组氨酸可提高肉鸡的饲料转化率,改善血浆抗氧化能力,增加肌肉中咪唑二肽含量。本试验条件下,饲粮添加1 000~2 000 mg/kg L-组氨酸较佳。从增加肌肉咪唑二肽含量考虑,饲粮L-组氨酸适宜添加量为1 276 mg/kg     

The solubilization of myofibrillar proteins of vertebrate skeletal muscle in water

Myofibrillar proteins of vertebrate skeletal muscles are insoluble in solutions of ionic strength that approximate physiological conditions. We established a method to solubilize more than 80% of chicken breast muscle myofibrillar proteins in water for the use of meat as a source of food protein. SDS‐polyacrylamide gel electrophoretic patterns of water‐soluble myofibrillar proteins demonstrated that all identified myofibrillar proteins except connectin/titin were soluble in water. A part of α‐actinin was released from myofibrils by repeated washing with 2.5 mmol/L NaCl and 5 mmol/L L‐histidine solution, and subsequent destruction of connectin/titin in washed myofibrils by ultrasonication resulted in solubilization of a large fraction of chicken breast muscle myofibrillar proteins in water. Myofibrillar proteins of chicken leg, pork loin, beef shoulder loin, and lamb were also solubilized in water using this procedure.     

Carnosine, anserine and taurine contents in individual fibres from the middle gluteal muscle of the camel

High muscle camosine and anserine contents contribute significantly to intra-cellular physico-chemical buffering. Our aim was to measure carnosine, anserine and taurine contents directly in individual type I, HA and IIB fibres from the middle gluteus muscle of the camel. Mean carnosine contents in type I, IIA and IIB were 24·6 ±9·2, 39·4 ±11·4 and 42·8 ±18·8 mmol kg⁻¹ dry weight (dw), respectively. Mean anserine contents in type I, HA and IIB fibres were 30·0 ±8·4, 37·3 ±10·1 and 34·5 ±9·7 mmol kg⁻¹ dw, respectively. Mean taurine contents in type I, IIA and HB fibres were 42·4 ±15-9, 203 ±12·9 and 24·7 ±15·9 mmol kg⁻¹ dw, respectively. Higher carnosine contents in type II fibres emphasise the importance of carnosine to intra-muscular acid-base regulation. A specific role for taurine in type I fibres is unclear.     

Histidine and carnosine reduce muscle fatigue in Xenopus laevis

The inclusion of either histidine or carnosine in a frog Ringer’s perfusate at a concentration of 20 mmol/dm³ reduced fatigue in the gastrocnemius muscle ofXenopus laevis significantly. Histidine was superior to carnosine in this respect. The evidence also suggests that muscles perfused with histidine exhibit superior recovery, accumulate less lactic acid and lose significantly less K⁺ from the intracellular fluid. The implications of these findings in future research on capture myopathy are discussed.     

Correlation between skeletal muscle fiber type and free amino acid levels in Japanese Black steers

Free amino acids are important components of tastants and flavor precursors in meat. To clarify the correlation between muscle fiber type and free amino acids, we determined the concentrations of various free amino acids and dipeptides in samples of different muscle tissues (n = 21), collected from 26‐month‐old Japanese Black steers (n = 3) at 2 days postmortem. The proportions of the myosin heavy chain (MyHC), slow (MyHC1) and fast (MyHC2) isoforms were determined by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE). The contents of free amino acids and dipeptides were measured by high performance liquid chromatography (HPLC). The MyHC isoform composition varied among the tissue samples. The MyHC1 proportion ranged from 6.9% ± 3.9% to 83.3% ± 16.7%. We confirmed that there was a strong positive correlation between MyHC1 composition and total free amino acid concentrations, including those for two dipeptides. Among the 31 measured free amino acids and dipeptides, 11 showed significant positive correlations and five showed significant negative correlations with MyHC1 composition. These results suggest that a high MyHC1 content induces high free amino acid contents in bovine muscles possibly because of greater oxidative metabolism. This high level of free amino acids could contribute to the intense flavor of meat that is rich in slow‐twitch fibers.     

Residue depletion of decoquinate in chicken tissues after oral administration

Wang, Z.‐Y., Cai, C.‐Y., Chang, X.‐H., Fei, C.‐Z., Qiu, M.‐Q., Jiang, S.‐X., Xue, F.‐Q., Zhang, L.‐F. Residue depletion of decoquinate in chicken tissues after oral administration. J. vet. Pharmacol. Therap.  36 , 116–121. A rapid, sensitive, and reliable high‐performance liquid chromatography tandem mass spectrometry (LC‐MS/MS) method was developed and validated for the analysis of decoquinate in chicken tissues. The compounds were extracted using acetonitrile by liquid–liquid extraction (LLE) and purified with an Oasis^? HLB solid‐phase extraction (SPE) cartridge. Chromatographic separation was performed on an XTerra C₁₈ reversed‐phase column with a mobile phase of water containing 0.1% formic acid and acetonitrile. The analyte was detected by tandem quadrupole mass spectrometry after positive electrospray ionization by multiple reaction monitoring. The detection and quantitation limits were 1 and 2.5 μg/kg, respectively. The recoveries of edible tissues ranged from 85.3% to 104.9%, with relative standard deviations (RSD) lower than 10.4%. The depletion profile of decoquinate was studied in healthy chickens after oral administration of feed containing 27.2 mg/kg decoquinate for 10 consecutive days. The residue concentrations of decoquinate in chicken muscle and liver were detected using the developed method. The highest residue concentrations were attained 0.25 day post‐treatment, and decoquinate residues were still detected 5 days postmedication in the tissues examined. The developed method has been successfully applied to the depletion study of decoquinate in chicken tissues. The recommended withdrawal period with oral administration based on our research is 3 days.     

Accurate analysis of taurine,anserine, carnosine and free amino acids in a cattle muscle biopsy sample

We have established an analysis method for some free amino acids (FAAs), as well as taurine (Tau), anserine (Ans) and carnosine (Car), in a fresh biopsy sample from cattle muscle. A series of model biopsy samples, corresponding to the mixtures of lean meat, fat and connective tissue, was prepared and showed high correlation coefficients between the compound concentration and the 3‐methylhistidine (3‐MeHis) content derived from hydrolysis of the biopsy sample (r = 0.74–0.95, P < 0.01). Interference from blood contamination could not be neglected, because the concentration of some FAAs in blood was comparable to that in muscle. However, it was possible to control the contamination of Tau, Ans, Car, glutamic acid, glutamine, asparatic acid and alanine to less than 5.0% when the blood contamination was controlled to less than 23%.These results suggest the necessity of measuring 3‐MeHis as an index of lean meat and hemoglobin as an index of blood contamination when compounds in muscle biopsy samples are evaluated. We have carried out a series of these analyses using one biopsy sample and reveal differences in Tau, Ans, Car and some FAAs in beef muscle after different feeding regimes.     

Interaction between the effects of evaporation rate and amount of simulated rainfall on development of the free-living stages of Haemonchus contortus

A factorial experiment (3 x 4 x 2 x 3) was conducted in programmable incubators to investigate interaction between the effects of rainfall amount, rainfall distribution and evaporation rate on development of Haemonchus contortus to L3. Sheep faeces containing H. contortus eggs were incubated on sterilised soil under variable temperatures typical of summer in the Northern Tablelands of NSW, Australia. Simulated rainfall was applied in 1 of 3 amounts (12, 24 or 32 mm) and 4 distributions (a single event on the day after deposition, or the same total amount split in 2, 3 or 4 equal events over 2, 3 or 4 days, respectively). Samples were incubated at either a Low or High rate of evaporation (Low: 2.1-3.4 mm/day and High: 3.8-6.1 mm/day), and faeces and soil were destructively sampled at 4, 7 and 14 days post-deposition. Recovery of L3 from the soil (extra-pellet L3) increased over time (up to 0.52% at day 14) and with each increment of rainfall (12 mm: <0.01%; 24 mm: 0.10%; 32 mm: 0.45%) but was reduced under the High evaporation rate (0.01%) compared with the Low evaporation rate (0.31%). All rainfall amounts yielded significantly different recoveries of L3 under Low evaporation rates but there was no difference between the 12 and 24 mm treatments under the High evaporation rate. The distribution of simulated rainfall did not significantly affect recovery of infective larvae. Faecal moisture content was positively associated with L3 recovery, as was the ratio of cumulative precipitation and cumulative evaporation (P/E), particularly when measured in the first 4 days post-deposition. The results show that evaporation rate plays a significant role in regulating the influence of rainfall amount on the success of L3 transmission.     

Effects of astaxanthin‐rich dried cell powder from Paracoccus carotinifaciens on carotenoid composition and lipid peroxidation in skeletal muscle of broiler chickens under thermo‐neutral or realistic high temperature conditions

Thirty‐two 15‐day old broiler chicks (Chunky strain ROSS 308) were randomly divided into four treatments in a 2 × 2 factorial design. The main factors were diet (basal diet or basal diet supplemented with 0.15% astaxanthin‐rich dried cell powder (Panaferd‐P [astaxanthin 30 ppm]) and ambient temperature (thermo‐neutral [25 ± 1°C] or high [35 ± 1°C for 6 hr]). Dietary supplementation with Panaferd‐P did not affect growth performance, though high ambient temperature decreased feed intake and the weight of breast tender muscle, liver, and heart. High ambient temperature also decreased redness in both breast and leg muscles of chickens, while Panaferd‐P increased redness and yellowness of breast and leg muscles of chickens. Panaferd‐P increased Paracoccus carotinifaciens‐derived pigments (i.e., adonixanthin, astaxanthin, adonirubin, and cantaxanthin) as well as corn‐derived pigments such as zeaxanthin and lutein in breast and leg muscles. High ambient temperature increased the malondialdehyde (MDA) concentration in breast muscle, while Panaferd‐P decreased the MDA concentration in breast muscle under both temperature conditions. Our results suggest that dietary supplementation with Panaferd‐P increases muscle carotenoid content, the redness and yellowness of meat and decreases the muscle MDA concentration in broiler chickens kept under thermo‐neutral or high ambient temperature conditions.     

β-丙氨酸的生理功能及其在动物生产中的应用

β-丙氨酸是一种不参与蛋白质合成的氨基酸,为肌肽、鹅肌肽等肌源活性肽合成的前体物,作为增强肌肉耐力的运动营养补充剂已经广泛应用于临床营养。研究表明,β-丙氨酸可提高动物生产性能,调控肌肉生长和肌源活性肽含量,改善肉品质量。本文旨在对β-丙氨酸的来源和代谢、生理功能及其在动物生产领域的应用进行综述,为β-丙氨酸在动物营养调控和生产实践中的应用提供理论依据。