The effect of litter size and day of lactation on amino acid uptake by the porcine mammary glands

Twelve multiparous sows (PIC Camborough 15; parity >2) were used to investigate the relationship between litter size and day of lactation, and plasma amino acid (AA) arteriovenous differences (A-VD), AA uptake, and plasma flow across the mammary glands. Sows were assigned randomly to one of the following litter sizes: 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 pigs per litter by cross fostering on d 2 postpartum. All sows were surgically fitted with catheters in the carotid artery and the main mammary vein. Matched arteriovenous blood samples were obtained on d 9, 12, 15, 18, 21, and 24 postpartum. Daily mammary uptake of AA was based on the product of plasma A-VD and daily mammary plasma flow (MPF). Daily MPF was estimated using the Fick method based on lysine conservation across the gland, and daily milk production. For the majority of AA, as litter size increased, A-VD did not increase, except for alanine (P < 0.05, linear and quadratic) and valine (P < 0.1; trend; linear and quadratic). As day of lactation increased, A-VD for the majority of AA increased (P < 0.05, linear and quadratic) except for arginine, lysine, and phenylalanine. As litter size increased, net daily mammary AA uptake increased for all indispensable AA (P = 0.001 to P < 0.05, linear and quadratic), excepting arginine. Milk production increased with increasing litter size (P < 0.001, linear) and with increasing day of lactation (P < 0.05, quadratic). Daily MPF increased (P < 0.05, linear) with increasing litter size, but did not change during the period measured from d 9 to 24. In conclusion, litter size appears to be a major determinant of net mammary AA uptake with daily mammary plasma flow a driving variable, whereas AA A-VD is a function of day of lactation and a major variable in determining net AA uptake with advancement of lactation.     

Effect of gestation dietary methionine-to-lysine ratio on methionine metabolism and antioxidant ability of high-prolific sows

The uptake and metabolism of methionine (Met) are critical for epigenetic regulation, redox homeostasis, and embryo development. Our previous study showed that appropriate supplementation of dietary Met promoted the birth weight and placental angiogenesis of high-prolific sows. To further explore the metabolic effect of Met on pregnant sows, we have evaluated the influence of dietary Met level on Met metabolism, and the relationship between metabolites of Met and reproductive performance, antioxidant ability, and placental angiogenesis throughout the gestation of high-prolific sows. Sixty sows (the 3rd parity, Large White) were randomly divided into 5 groups that were fed diets with standardized ileal digestible (SID) methionine-to-lysine (Met:Lys) ratios of 0.27 (control), 0.32, 0.37, 0.42, and 0.47 from the mating day (gestational d 0, G0d) until the farrowing day. HPLC-MS/MS analysis was used to simultaneously evaluate the metabolites related to Met, e.g. S-adenosylmethionine (SAM), S-adenosylhomocysteine (SAH), homocysteine (Hcy), cysteine (Cys), and glutathione (GSH). The concentration of SAM and SAH in plasma had significant fluctuations, especially in late pregnancy. Increasing dietary Met supplementation significantly improved the plasma SAM and methylation potential (SAM-to-SAH ratio) at d 114 of pregnancy (G114d). Moreover, a positive association of the plasma SAM concentration at G114d was observed with the litter weight of born alive (P < 0.05; R² = 0.58). Furthermore, Hcy concentration in plasma was at the lowest level for 0.37 ratio group at G114d. However, it significantly increased during late pregnancy. Moreover, there were negative correlations between plasma Hcy concentration at G114d (P < 0.05) and the placental vascular density in the fold and stroma (P < 0.05). Compared with the control group, the expression of vascular endothelial growth factor-A (VEGF-A) in the placenta tissue of 0.37 ratio group increased significantly (P < 0.05). Collectively, these findings indicate that dietary Met:Lys ratio (0.37 to 0.57) in the pregnant diet dose not influence the antioxidant ability of the high-prolific sows; however, the improvement of fetal development and placental angiogenesis of high-prolific sows by supplementation of Met are closely associated to the key Met-related metabolite of SAM and Hcy, respectively.     

Protein (lysine) restriction in primiparous lactating sows: effects on metabolic state,somatotropic axis,and reproductive performance after weaning

Low protein intake during lactation has been demonstrated to increase the loss of body protein and to reduce the reproductive performance of female pigs. The objectives of the current experiment were 1) to determine whether protein (lysine) restriction alters levels of somatotropic hormones, insulin, follicle-stimulating hormone, and leptin around weaning, and 2) to evaluate the relationships between these eventual alterations and postweaning reproductive performance. One day after farrowing, crossbred primiparous sows were randomly allocated to one of two diets containing 20% crude protein and 1.08% lysine (C, n = 12) or 10% crude protein and 0.50% lysine (L, n = 14) during a 28-d lactation. Diets provided similar amounts of metabolizable energy (3.1 Mcal/kg). Feed allowance was restricted to 4.2 kg/d throughout lactation, and litter size was standardized to 10 per sow within 5 d after farrowing. Catheters were fitted in the jugular vein of 21 sows around d 22 of lactation. Serial blood samples were collected 1 d before (day W - 1) and 1 d after (day W + 1) weaning, and single blood samples were collected daily from weaning until d 6 postweaning (day W + 6). Sows were monitored for estrus and inseminated. They were slaughtered at d 30 of gestation. During lactation, litter weight gain was similar among treatment groups. Reduced protein intake increased (P < 0.001) sow weight loss (-30 vs -19 kg) and estimated protein mobilization throughout lactation (-4.1 vs -2.0 kg). On day W - 1, L sows had higher (P < 0.02) plasma glutamine and alanine concentrations, but lower (P < 0.05) plasma tryptophan and urea than C sows. Mean and basal plasma GH were higher (P < 0.001), whereas plasma IGF-I and mean insulin were lower in L than in C sows on day W - 1. Preprandial leptin did not differ between treatments on day W - 1, but was higher (P < 0.01) in L sows than in C sows on day W + 1. Mean FSH concentrations were similar in both treatments on day W - 1 (1.3 ng/mL), but L sows had greater (P < 0.001) mean FSH on day W + 1 than C sows (1.6 vs 1.2 ng/mL). The weaning-to-estrus interval (5 +/- 1 d) was similar in both groups. Ovulation rate was lower in L than in C sows (20.0 +/- 1 vs 23.4 +/- 1, P < 0.05). No obvious relationships between reproductive traits and metabolic hormone data were observed. In conclusion, these results provide evidence that protein (lysine) restriction throughout lactation alters circulating concentrations of somatotropic hormones and insulin at the end of lactation and has a negative impact on postweaning ovulation rate.     

Effect of nutrient intake on mammary gland growth in lactating sows

Sixty-one primiparous sows were used to determine the response of mammary gland growth to different energy and protein intakes during lactation. After birth, litter size was set to 9 or 10 pigs. Sows were slaughtered at selected times up to 30 d of lactation. Individual sows were fed one of four diets that were combinations of different amounts of energy and protein (3.0 Mcal ME and 8.0 g lysine/kg diet; 3.0 Mcal ME and 16.2 g lysine/kg diet; 3.5 Mcal ME and 6.4 g lysine/kg diet; or 3.5 Mcal ME and 13.0 g lysine/kg diet). Mammary glands were collected at slaughter and trimmed of skin and the extraneous fat pad. Each gland was weighed, cut in half to measure cross-sectional area, ground, and stored at -20 degrees C for chemical analysis. Frozen, ground tissue was used to determine dry matter, dry fat-free tissue (DFFT), total tissue protein, ash, and DNA content. Only glands known to have been suckled were included in this data set. Response surface regression was used for statistical analysis. The percentage of protein, fat, ash, and DNA in each suckled mammary gland was affected only by total energy intake (P<.05). The percentage of dry tissue and fat decreased as the total energy consumed during lactation increased, whereas the percentage of protein and DFFT increased as total energy intake increased. There were quadratic effects (P<.05) of both total energy and protein intake on wet weight, dry weight, protein amount, DFFT amount, and DNA amount of each suckled mammary gland during lactation. This study shows that mammary gland growth is affected by nutrient intake during lactation. The weight of suckled mammary glands and the amount of mammary tissue protein, DFFT, and total DNA were maximal on d 27.5 of lactation when sows had consumed an average of 16.9 Mcal of ME and 55 g of lysine per day during lactation. Provision of adequate amounts of nutrients to sows during lactation is important for achieving maximal growth of mammary glands and maximal milk production.     

Effect of intraperitoneal administration of lysine and methionine on mohair yield and quality in Angora goats.

Eight mature Angora wethers (average BW 47.2 kg) were used in a 4 x 4 replicated Latin square design to evaluate responses to intraperitoneal (IP) administration of amino acids. The IP treatments consisted of saline (Control), methionine 1 g/d (Met), lysine 2 g/d (Lys), and methionine + lysine (Met + Lys). The amino acids dissolved in Control were infused continuously for the first 14 d of each 28-d period using peristaltic infusion pumps. Average d-28 grease and clean mohair yields (grams/100 square centimeters), and fiber diameter (micrometers) and length (centimeters) measurements during Control administration were 8.8, 7.6, 40, and 2.3, respectively. Mean clean mohair yield and fiber diameter increased by 5.3% (P less than .039) and 2.5% (P less than .067), respectively, with Met administration but were decreased by 9.2% (P less than .033) and 3.8% (P less than .001), respectively, by Lys administration; however, mean fiber length was increased (P less than .014) 21.7% by Lys infusion. The goats did not exhibit increased grease (P greater than .939) and clean (P greater than .477) mohair yields and fiber diameter (P greater than .619) when treated with Met+Lys. A N balance trial was conducted during d 10 through 14 of each period. Total retained N (grams per day) during Control administration was 18.4 and 24% greater than Met (P greater than .281) and Lys (P less than .061), respectively. When expressed as a proportion of N intake, retained N was lowest (P less than .127) in the Lys infusion group. Jugular blood ammonia N and plasma glucose and total protein concentrations were not affected (P greater than .10) by treatment.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of ambient temperature on mammary gland metabolism in lactating sows

Two groups of three multiparous Large White x Landrace sows were used to investigate the direct effect of ambient temperature on mammary gland metabolism. Sows from the first group were exposed to temperatures of 28 degrees C between d 8 and 14 of lactation, and 20 degrees C between d 15 and 21; treatments were reversed in the second group. Four to six d after farrowing, an ultrasonic blood flow probe was implanted around the right external pudic artery and catheters were fitted in the right anterior mammary vein and in the carotid artery. After surgery all sows were fed 3.8 kg/d of a lactation diet. The arteriovenous (AV, mg/L) plasma samples were obtained every 30 min between 0915 and 1545 on d 5 of exposure to ambient temperature; the same day, milk samples were collected at 1630. Additional arterial samples were obtained between 1000 and 1100 on d 1, 4, and 6 of exposure. Milk yield was estimated from the body weight gain of the litter. Elevated temperature tended to reduce BW loss (2.44 vs 1.82 kg/d, P < 0.10), but did not affect milk yield (11.0 kg/d). Glucagon and leptin arterial concentrations increased (12 and 8%, respectively; P < 0.10), but thyroxin and triiodothyronine concentrations decreased (26 and 16%, respectively; P < 0.01) between 20 and 28 degrees C. Expressed as a percentage of total nutrients, AV difference, glucose, amino acids, triglycerides (TG), free fatty acids, and lactate A-V differences represented 60, 20, 11, 8, and 1%, respectively. Exposure to 28 degrees C increased the extraction rate of glucose, TG, and a-amino acid N (13, 8, and 2.5%, respectively; P < 0.10). The extraction rates of essential and nonessential amino acids were not affected by temperature. The right pudic artery mammary blood flow increased (872 vs 945 mL/min, P < 0.05) between 20 and 28 degrees C, whereas milk yield was unaffected by temperature. It is suggested that this apparent inefficiency of the sow mammary gland in hot conditions could be related to an increase of proportion of blood flow irrigating skin capillaries in order to dissipate body heat.     

日粮蛋氨酸及赖氨酸水平对雌性肉仔鸡胴体组成的影响

本试验旨在探讨日粮蛋氨酸和赖氨酸水平对 3～ 6周龄艾维茵母鸡胴体组成及脂肪代谢的影响。试验采用 3× 5双因子试验设计 ,蛋氨酸设 3个水平 ( 0 32 % ,0 4 4% ,0 56 % ) ,赖氨酸设 5个水平( 0 7% ,0 85% ,1 0 % ,1 1 5% ,1 30 % ) ,一共形成 1 5个处理组。基础日粮由玉米、豆粕、花生粕、玉米蛋白粉等组成。含代谢能 1 2 73MJ/Kg ,粗蛋白 1 8 8% ,赖氨酸 0 7% ,蛋氨酸 0 32 %。试验末分别检测生产性能 ,屠宰性能及血液生化指标。结果表明 :日粮赖氨酸和蛋氨酸显著影响了肉仔鸡的胴体组成和脂肪代谢 (P <0 0 1 )。赖氨酸和蛋氨酸缺乏或过量 ,血浆尿酸、甘油三脂和极低密度脂蛋白浓度均呈上升趋势。日粮赖氨酸 ( 0 70 % )和蛋氨酸 ( 0 32 % )缺乏显著降低了肉仔鸡的体增重、胸肉率及全净膛率 (P<0 0 5) ,而腹脂沉积偏高。适度提高日粮赖氨酸和蛋氨酸水平可以提高全净膛率 ,胸肌率 ,降低腹脂沉积 ,从而改善胴体品质     

Measurement of blood flow through the mammary gland in lactating sows: methodological aspects

Two replicates of three multiparous crossbred Large White x Landrace lactating sows were used to develop a technique for the continuous direct measurement of the blood flow through the mammary gland using transit time ultrasound. Four to six days after farrowing, an ultrasonic transit time flow probe was implanted around the right external pudic artery in order to measure the short-term variations of mammary blood flow through this vessel in response to postural change (standing vs lying), meal distribution, hand-milking, and weaning. After surgery, all sows were fed 3.8 kg/d of a lactation diet and housed either at 20 or 28 degrees C. The implantation of the ultrasonic blood flow probe was successful in all six operated sows. Postmortem examination did not indicate the presence of infection, any collateral bypassing the flow probe, or a reduction of artery diameter. The right pudic artery mammary blood flow (PMBF) was measured for 8.5 h over two periods of three days (d 11 to d 13 and d 18 to d 20 of lactation). The PMBF averaged 910 +/- 238 mL/min but was variable within 1 d. Compared with the lying position, PMBF was decreased (- 6%, P < 0.05) when sows were standing. Between 0 to 15 and 16 to 30 min after oxytocin injection (t = 0) and hand-milking, PMBF remained constant (P = 0.05; 801 vs 767 mL/ min) and increased (P = 0.02), respectively, in comparison with the mean calculated over the preceding 30-min period (982 vs 784 mL/min). The PMBF increased (P < 0.05) after meal distribution and reached a peak 65 min later (i.e., 980 mL/min). The PMBF decreased regularly after separation of piglets at weaning; at 8 and 16 h after weaning, PMBF was 60 and 40% of the value recorded before weaning, respectively. Assuming that PMBF drains one-quarter of the whole mammary gland, it can be calculated that blood flow through the entire mammary gland averages 3.6 L/min and that about 470 L of blood are required to produce 1 kg of milk. The proposed methodology constitutes a new technique to measure direct mammary blood flow and its short-term factors of variation.     

Triennial Lactation Symposium: Mammary metabolism of amino acids in dairy cows

Although in dairy cows the mammary gland (MG) is the major net user of essential AA (EAA) supply, milk protein synthesis from absorbed EAA is not a straightforward process. Early studies identified 2 groups of EAA based on different pattern of mammary utilization: group 1 [Met, Phe (+Tyr), Trp], where MG uptake was similar to secretion in milk protein, and group 2 (Arg, Ile, Leu, Lys, Thr, and Val), where uptake exceeded milk protein output. This review examines the validity of this classification under variable protein supply through a meta-analysis, with the outcomes then explained with studies in which the fates of individual EAA were monitored using isotope approaches. For the meta-analysis, the Fick principle, based on stoichiometric transfer of Phe+Tyr uptake to milk protein, was used to estimate mammary plasma flow across all studies. This approach was judged acceptable because doubling Phe supply did not result in mammary oxidation of Phe+Tyr and either limited or no contribution of peptides to Phe and Tyr mammary supply could be detected. The AA content of proteins synthesized by the MG was estimated from milk protein composition, and the uptake-to-output ratio (U:O) for individual AA was re-calculated based on these assumptions. Analysis of individual samples by isotopic dilution resulted in reduced variance compared with analysis on pooled samples performed with an AA analyzer. Globally, the U:O of His and Met is maintained close to unity under variable protein supply. The group 2 AA could be subdivided. First, the U:O for group "2v" AA (Ile, Leu, Val, and Lys) is greater than 1 and varied with protein supply. Accordingly, the increased U:O of Leu, induced by duodenal casein infusion, led to extra-mammary Leu oxidation. Decreasing Lys supply decreased Lys U:O and the associated transfer of N to non-EAA, mainly to Glx, Asx, Ser, and Ala. Second, the U:O of group "2nv" AA, Arg and Thr, does not vary with protein supply. The Arg U:O averages 2.5, whereas the Thr U:O, albeit averaging 1.2, does not differ from unity. Excess of both these AA is probably directed toward the synthesis of non-EAA rather than energy supply. Overall, the ability of the MG to use excess EAA-N supply offers alternative sources of N and C for energy provision, lactose synthesis and non-EAA synthesis. The latter function spares dietary non-EAA for other necessary processes, such as gluconeogenesis and energy supply, in other tissues to support lactation.     

50～75日龄番鸭日粮限制性氨基酸水平研究

本试验旨在研究日粮不同赖氨酸粗蛋白质比例和蛋氨酸(含硫氨基酸)赖氨酸比例对50~75日龄番鸭生产性能、血清生化指标和肌肉品质的影响。试验选用1440只健康50日龄番鸭,随机分为9组,每组4个重复,每重复40只鸭,饲喂9种不同赖氨酸粗蛋白质比例(Lys/CP)(分别为5%、5.5%、6%)和蛋氨酸(含硫氨基酸)赖氨酸比例(Me(SAA)t/Lys)(分别为35%(75%)、40%(80%)、45%(85%))的玉米-豆粕型日粮。试验结束时,进行称重,每个重复挑选2只接近平均体重的番鸭进行采血和屠宰。结果显示:①Lys/CP对日均采食量(ADFI)、日均增重(ADG)影响显著,5%和5.5%组显著高于6%组(P<0.05);Me(tSAA)/Lys为40%(80%)的组ADFI显著高于45%(85%)组,40%(80%)组ADG显著高于其他两组,而料肉比显著低于其他两组;②Lys/CP为5%的组血清尿酸和甘油三酯含量显著高于(P<0.05)其他两组;Me(SAA)t/Lys对血清尿酸和甘油三酯含量影响不显著(P>0.05);日粮Lys/CP和Me(tSAA)/Lys对血清甘油三酯含量存在交互作用;③日粮Lys/CP为5.5%的组胸肌干物质(DM)含量显著高于其他两组,粗蛋白质(CP)含量显著高于6%组;Me(tSAA)/Lys为35%(75%)和40%(80%)的组胸肌CP含量显著高于45%(85%)组;日粮Lys/CP和Me(tSAA)/Lys对胸肌DM和CP含量存在交互作用;④日粮Lys/CP为5.5%和6%的组腿肌DM含量显著高于5%组,5.5%组腿肌CP含量显著高于其他两组;Me(tSAA)/Lys为40%(80%)和45%(85%)的组腿肌DM含量显著高于35%(75%)组;日粮Lys/CP和Me(tSAA)/Lys对腿肌DM和CP含量存在交互作用。     

Estimation of the standardized ileal digestible lysine requirement for primiparous pregnant sows

This experiment was conducted to determine the optimal standardized ileal digestible lysine (SID Lys) level in diets fed to primiparous sows during gestation. A total of 150 (Landrace × Large White) crossbred gilts (weighing 149.9 ± 3.1 kg) were fed gestation diets (12.55 MJ of ME/kg) containing SID Lys levels of 0.43, 0.52, 0.60, 0.70 or 0.80% respectively. Gilts were fed 2.0 kg/day from day 1 to 80 and 3.0 kg/day from day 80 to 110 of gestation respectively. Gilts were allocated to treatments based on their body weight on the day of breeding. Weight gain from day 80 to 110 increased with increasing dietary SID Lys levels (p = 0.044). Fitted broken‐line (p = 0.031) and quadratic plot (p = 0.047) analysis of body weight gain indicated that the optimal SID Lys level for primiparous sows was 0.70 and 0.69% respectively. During gestation, neither backfat thickness nor loin eye area was affected by dietary SID Lys level. Increasing dietary Lys had no effect on the litter size at birth or pigs born alive per litter. Litter weight at birth was not affected by dietary SID Lys level. The litter weight variation at birth quadratically decreased with increasing dietary SID Lys (p = 0.021) and was minimized at 0.70% dietary SID Lys. Gilts fed the 0.70% SID Lys diet had the highest dry matter (p = 0.031) and protein (p = 0.044) content in colostrum. On day 110 of gestation, gilts fed the 0.70% SID Lys diet tended to have the highest serum prolactin (p = 0.085) and serum insulin (p = 0.074) levels. The data demonstrate that the optimal dietary SID Lys was 0.70% for pregnant gilts, which is similar to the recommendation of 0.69% that was estimated by the NRC (2012).     

不同生理阶段犊牛赖、蛋、苏氨酸平衡模式研究

应用部分扣除饲粮氨基酸(AA)的氮平衡试验法研究了中国荷斯坦犊牛不同生理阶段(2～3周龄和5～6周龄)赖(Lys)、蛋(Met)、苏氨酸(Thr)平衡模式。每个阶段各用12头犊牛进行N平衡试验。每个N平衡试验均设4个处理组,即氨基酸相对平衡的代乳品(PC)组(赖氨酸、蛋氨酸、苏氨酸含量依次为2.34%、0.72%、1.80%)、在PC基础上轮流将赖、蛋、苏氨酸扣除30%的其它3种代乳品(分别为PC-Lys、PC-Met、PC-Thr)组。每个处理设3个重复,每个重复1头牛。所有代乳品等能(17.8MJ/kg)等蛋白(22%)。试验结果表明,按单位代谢体重的N沉积与氨基酸摄入量计算,中国荷斯坦犊牛2～3周龄日粮模式为代乳品时,Lys、Met、Thr的平衡比例100:29:70;5～6周龄日粮模式为代乳品、开食料和羊草时,平衡比例为100:30:60;两阶段3种氨基酸的限制性顺序均为Lys、Met、Thr。     

Mammary gland growth as influenced by litter size in lactating sows: impact on lysine requirement

Twenty-eight primiparous sows were used to determine the effect of litter size on the growth of mammary glands and nursing pigs during lactation. Litter size was set to 6, 7, 8, 9, 10, 11, or 12 pigs by cross-fostering immediately after birth. Four sows were allotted to each litter-size group. Sows were allowed to consume a daily maximum of 13.6 Mcal ME and 46.3 g of lysine during lactation. Sows were slaughtered on d 21 (20.6+/-1.1) of lactation. Mammary glands were collected at slaughter and trimmed of skin and the extraneous fat pad. Each gland was separated, weighed, and ground for chemical analysis. Dry matter, dry fat-free tissue (DFFT), crude protein, ash, and DNA contents were measured. Only glands known to have been nursed were included in the data set. Wet and dry weights and the amounts of DFFT, protein, DNA, ash, and fat in individual nursed mammary glands linearly decreased (P<.05) as litter size increased. Percentages of DFFT, protein, and DNA were quadratically affected (P<.05) by litter size on d 21 of lactation. Total mammary wet and dry weights and total DFFT, protein, DNA, fat, and ash amount of all nursed mammary glands of each sow were increased as litter size increased (P<.05). Changing litter size from 6 to 12 pigs resulted in 2,098, 432, 253, 227, 4.4, 178, and 20 g increases in the amounts of total mammary wet weight, dry weight, DFFT, protein, DNA, fat, and ash, respectively, on d 21 of lactation. Litter weight gain was 18.1 kg greater in sows with 12 pigs than in sows with 6 pigs. Sows with a larger litter size had a greater increase in total mass of mammary gland tissue and litter weight but had lower growth of individual nursed mammary glands and individual pigs than sows with the smaller litter size. The need for nutrients to support additional mammary gland and litter growth as litter size increases should be considered when estimating nutrient requirements for lactating sows. Sows need an additional .96 g lysine per day to account for mammary gland growth for each pig added to a litter.     

Lysine uptake by mammary gland tissue from lactating sows

Kinetic properties and substrate specificity of the lysine transport system in porcine mammary gland were studied using mammary tissue explants from nine lactating sows. Sodium dependence of lysine uptake was determined by replacing sodium in the medium with choline. Kinetic parameters of lysine uptake were determined using lysine concentrations from 5 microM to 5.12 mM. Competition of lysine uptake by other amino acids was determined using the cationic amino acids, arginine and ornithine, and using other essential amino acids. Transport of lysine was time-dependent and was unaffected by replacing sodium with choline. Lysine uptake occurred by a transport mechanism with a Km of approximately 1.4 mM and a Vmax of 7.9 mmol x kg cell water(-1) x 30 min(-1). Lysine uptake was inhibited by arginine and ornithine and by high concentrations of L-alanine, L-methionine, L-leucine, cycloleucine, and D-lysine, but not by 2-(methylamino)-isobutyric acid. This transport mechanism is the primary system responsible for uptake of cationic amino acids in lactating sow mammary tissue. The relatively high Km, compared with physiological blood concentrations of lysine, indicates that the kinetic properties of the lysine transport system should not be limiting to milk protein synthesis. Transmembrane transport of lysine by lactating sow mammary tissue should be a direct function of plasma concentrations. However, interactions of other amino acids with the uptake system may affect lysine uptake.     

牛乳腺上皮细胞SNAT2对氨基酸调节乳合成的影响

试验旨在研究氨基酸转运体钠离子依赖的中性氨基酸转运蛋白（the sodium-dependent neutral amino acid transporter 2,SNAT2）在牛乳腺上皮细胞（bovine mammary epithelial cell,BMEC）中对氨基酸调节乳合成的影响。利用组织块法成功培养原代BMEC,添加不同氨基酸（蛋氨酸（Met）、赖氨酸（Lys）和亮氨酸（Leu）刺激BMEC后,通过实时荧光定量PCR、Western blotting技术和甘油三酯试剂盒检测SNAT2、酪蛋白（β-casein）基因的表达量和BMEC培养液上清甘油三酯的分泌量;将N-flag-SNAT2真核表达载体及SNAT2 siRNA分别转入细胞中进行SNAT2基因的过表达和敲低试验;通过Western blotting和甘油三酯试剂盒分别检测SNAT2、哺乳动物雷帕霉素靶蛋白（the mammalian target of rapamycin,mTOR）、β-casein蛋白表达量和BMEC培养液上清的甘油三酯含量。结果显示,3种氨基酸（Met、Lys、Leu）均能显著促进BMEC分泌乳蛋白和乳脂,并激活mTOR信号途径,其中Met、Lys还能够显著上调SNAT2基因表达;SNAT2能够正向调节BMEC乳蛋白和乳脂肪的合成,并激活mTOR信号通路,说明氨基酸激活mTOR信号通路是通过SNAT2基因介导完成的,进而调节了BMEC乳蛋白和乳脂肪合成。     

Dietary protein concentration affects plasma arteriovenous difference of amino acids across the porcine mammary gland

The objective of this study was to determine whether the porcine mammary gland responds to increasing dietary CP concentration through changes in AA arteriovenous difference (a-v). Sixteen Landrace x Yorkshire lactating sows were provided ad libitum access to one of four isocaloric diets varying in CP concentration (7.8, 13.0, 18.2, and 23.5 %; as-fed basis). Litters were adjusted to 11 pigs within 48 h of birth. Sows were fitted with catheters in the carotid artery and main mammary vein on d 4. On d 10, 14, 18, and 22 of lactation, arterial and venous blood samples were obtained every 30 min over 6 h. Milk yield was estimated on d 11 and 21 using the D2O dilution technique. Final litter sizes on d 21 were 10.3, 11, 9.5, and 11 piglets for sows fed the 7.8, 13.0, 18.2, and 23.5% CP diets, respectively. Piglet ADG tended (P = 0.088) to increase with increasing dietary CP concentration and were 186, 221, 220, and 202 g for sows fed the 7.8, 13.0, 18.2, and 23.5% CP diet, respectively. Daily total milk yield on d 21 (kg milk/d) tended (P = 0.099) to increase, and average milk yield per nursed piglet (kg of milk-pig(-1)d(-1)) increased (P < 0.05) with increasing CP concentration and were, on a per-piglet basis, 0.95, 1.19, 1.14 and 1.13 kg of milk/d for the 7.8, 13.0, 18.2, and 23.5% CP diets, respectively. As dietary CP increased from 7.8 to 23.5%, isoleucine and leucine a-v increased linearly only (linear, P < 0.01); all other AA a-v increased, reached a maximum in sows fed 18.2% CP, and decreased thereafter in sows fed 23.5% CP (quadratic, from P = 0.10 to P < 0.05). Amino acid uptake by the entire udder and by each gland increased (linear, P < 0.05) with increasing dietary CP. Arteriovenous differences response to increasing day of lactation varied among AA, from no change for histidine, isoleucine, lysine, methionine, tryptophan, and valine, to a linear trend increase for arginine (P = 0.055), leucine (P = 0.064), phenylalanine (P = 0.101), and threonine (P = 0.057). In summary, for the majority of AA, a-v increased with increasing dietary CP concentration from 7.8 to 18.2%, but decreased when CP concentration exceeded 18.2%. In contrast, mammary AA uptake, piglet ADG and milk yield per pig increased linearly with increasing dietary CP, suggesting a coordinated regulation between AA delivery and transport to meet the demand for milk yield.     

The effect of different dietary ratios of lysine,arginine and methionine on biochemical parameters and hormone secretion in turkeys

We postulated that the use of optimal levels and proportions of Lys, Arg and Met in compound feed allows for optimal exploitation of the growth potential of contemporary slaughter turkey hybrids and reduces metabolic disorders. The aim of the study was to determine the effect of different proportions of Lys, Arg and Met in diets whose Lys content is in accordance with NRC recommendations, that is a low level, on selected parameters of protein, lipid and carbohydrate metabolism and on hormone secretion in turkeys. The lowest Arg content (90% Lys) in the diet resulted in an increase in plasma total cholesterol levels in the turkeys as compared to higher Arg content (100% or 110% of Lys), (2.50 vs. 2.09 vs. 1.83). Plasma HDL and creatinine concentration increased in turkeys fed diets with higher Arg content (100% and 110% Lys) compared to turkeys receiving the diet with the lowest Arg content (90% Lys). Compared to turkeys receiving the lowest and intermediate Arg content (90% and 100% Lys), the diet with the highest content of this AA (110% Lys) resulted in an increase in the plasma T4 level (71.21 vs. 86.60 vs. 128.2). The varied Arg and Met levels relative to Lys did not affect the secretion of neurotransmitters or hormones regulating glucose metabolism. At low levels of Met in the diet, an decrease in Arg relative to Lys from 100% to 90% caused a growth depression of turkeys (10.68 vs. 10.21 kg), which was not noted in the case of the higher Met content. When using the Lys level recommended by NRC in the turkey diet, the optimal Arg level is 100% and Met is 45% compared to Lys.     

Mammary arteriovenous differences of glucose, insulin, prolactin and IGF-I in lactating sows under different protein intake levels

Mammary uptake of nutrients is dependent on their availability in the circulation but the role of hormones in that process is not known. Arteriovenous differences (AVD) of glucose and key hormones across the mammary glands were therefore determined in sows fed varying levels of protein. Sixteen lactating sows (four/dietary treatment) were fed a 7.8, 13.0, 18.2 or 23.5% crude protein (CP) isocaloric diet throughout lactation and their litters were standardized to 11 pigs within 48 h of birth. The anterior main mammary vein and a carotid artery were cannulated on day 4+/-1 of lactation and blood samples were collected every 30 min over 6h on days 10, 14, 18 and 22 of lactation to measure glucose, insulin, IGF-I, and prolactin (PRL) concentrations. Amino acid data from these sows were previously published and used here to determine residual correlations. Dietary treatments had no effect on any of the insulin or PRL variables measured (P>0.1) and, on day 18 only, IGF-I AVD was greater (P=0.05) for sows on the 23.5% compared to the 18.2% diet. On days 18 and 22, sows fed the 13% CP diet had greater arterial, venous and AVD glucose concentrations than sows fed other diets (P<0.05). Total arterial amino acid concentrations were correlated to arterial insulin (P<0.001) and PRL (P<0.05) concentrations, but not to those of IGF-I (P>0.1). Mammary AVD for total (P<0.001) and essential amino acids (P<0.05) were correlated to arterial concentrations of insulin, but not to those of IGF-I (P>0.1) or PRL (P>0.1). Mammary AVD of both total (P<0.01) and essential (P<0.05) amino acids were also correlated to mammary PRL AVD. In conclusion, dietary protein level did not affect mammary AVD and circulating lactogenic hormone concentrations. Yet, amino acid utilization by the sow mammary gland seems to be regulated via both circulating insulin concentrations and PRL binding to and uptake by porcine mammary cells.     

赖氨酸、蛋氨酸和苏氨酸对犊牛生长性能和血清生化指标的影响

本试验旨在研究赖氨酸、蛋氨酸和苏氨酸对犊牛生长性能和血清生化指标的影响.选取24头新生荷斯坦犊牛进行8周饲养试验,按照体重和出生时间相近原则分为4组,采用4种氨基酸模式的代乳粉,分别为氨基酸平衡组(PC组)、扣除赖氨酸组(PC-Lys组)、扣除蛋氨酸组(PC-Met组)、扣除苏氨酸组(PC-Thr组),扣除量均为对照组...     

30～38周龄产蛋鸡理想氨基酸模式的研究

本试验旨在通过4个剂量-反应试验,研究30～38周龄产蛋鸡对赖氨酸(Lys)、蛋氨酸(Met)、色氨酸(Trp)和苏氨酸(Thr)4种必需氨基酸(AA)的需要量及其比例.每个试验均采用360只海兰灰产蛋鸡,设5个处理,分别采食5个AA梯度水平的试验饲粮,每个处理6个重复,每个重复12只鸡.采用蛋公鸡回肠食糜法测定饲料原料的标准回肠可消化(SID)值.试验预试期14 d,正试期56 d.结果表明:海兰灰产蛋鸡的产蛋率、蛋重、日产蛋量和料蛋比与SIDLys、SID Met、SID Trp和SID Thr的摄入水平均有较好的折线回归关系;以日产蛋量为自变量(x),AA摄入水平为依变量(Y)分别建立的回归方程为:YSID Lys=51.60-0.041 6(682.5-x),R2=0.96;YsID Met=51.43-0.085(314.1-x),R2 =0.95;YSID Trp =51.17-0.155 4(163.0-x),R2=0.97;YsID Thr=52.85-0.057 6(499.5-x),R2=0.93.在饲粮粗蛋白质12.8％的条件下,以日产蛋量为参照指标确定的30 ～ 38周龄产蛋鸡的SID Lys、SID Met、SID Trp和SID Thr需要量分别为684、313、171和506 mg/(只·d).以日产蛋量为指标建立的30～38周龄产蛋鸡理想SID AA模式为:Lys:Met:Trp:Thr=100∶46∶24∶73.