Lys Concentration Research Articles

Optimal lysine in diets for high-yielding lactating sows1.

The objective of the current study was to determine the optimal concentration of dietary standardized ileal digestible (SID) Lys required to maximize litter gain and minimize sow BW loss in modern high-yielding lactating sows when SID CP was kept constant across dietary treatments. A total of 396 parity 1 to 5 sows (L × Y, DanBred, Herlev, Denmark) were included in the study from day 3 after farrowing until weaning at day 26. Sows were allocated to 6 dietary treatments increasing in SID Lys concentration (6.19, 6.90, 7.63, 8.33, 9.04, and 9.76 g/kg). Diets were isoenergetic (14.04 MJ ME/kg as-fed). Litters were standardized to 14 piglets at day 3 ± 2 d postpartum. At day 3 ± 2 d and at day 26 ± 3 d in lactation, litter weight, and sow BW and back fat were registered. On a subsample of 72 parity 2 to 4 sows, litters were additionally weighed at days 10 and 17 ± 3 d, and milk and blood were sampled at day 3 ± 2 d, and 10, 17 and at 24 ± 3 d in lactation. For the 72 sows, body pools of fat and protein were also determined at days 3 ± 2 and 26 ± 3 d using the D2O dilution technique. All data were analyzed as a randomized complete block design using PROC MIXED in SAS. Furthermore, data were subjected to linear and quadratic polynomial contrasts. Variables with quadratic or linear effects or days in milk × treatment interactions were selected for analysis in PROC NLMIXED using linear broken-line models to evaluate optimal SID Lys concentrations. Only models that converged and the best fitting models were included. Average daily litter gain increased until a breakpoint at 8.11 g/kg of SID Lys (as-fed). At and above the breakpoint, multiparous and primiparous sows had litter gains of 3.36 and 2.93 kg/d, respectively. Weaning litter size (13.0 ± 0.1) was similar between the 6 dietary treatments (P = 0.28). Lactation sow BW loss was minimized to 0.17 kg/d at 9.05 g/kg of SID Lys and sow body protein loss was minimized to 0.23 kg at 9.22 g/kg of SID Lys. Linear broken-line analyses showed that for 3, 10, 17, and 24 DIM, plasma urea was minimized at 7.02, 8.10, 8.73, and 8.32 g/kg of SID Lys, respectively, and milk fat was maximized at 7.80 g/kg of SID Lys. In conclusion, in our conditions, high-yielding lactating sows required 8.11 g/kg of SID Lys to maximize litter gain and 9.05 g/kg of SID Lys to minimize sow BW loss. Based on plasma urea, the optimal dietary concentration of SID Lys was lowest in week 1, intermediate in week 2 and 4, and greatest in week 3 of lactation.

Effects of varying extracellular amino acid profile on intracellular free amino acid concentrations and cell signaling in primary mammary epithelial cells

Extracellular amino acid profiles affect intracellular AA concentrations and profile as well as signaling proteins that regulate protein translation rates. The objective of this study was to assess whether various extracellular AA profiles and varied ratios of Lys to Met would increase the phosphorylation of signaling proteins related to protein metabolism. Six AA profiles, reflecting Dulbecco's modified Eagle's medium (DMEM), blood meal (BM), corn gluten meal (CGM), casein (CAS), plasma of lactating cows (PLA), and a negative control (NEG) represented the first factor (F1), and the ratio of Lys to Met (unaltered or set to 3:1) was the second factor (F2). Treatments were arranged in a 6 × 2 factorial manner, resulting in 12 treatments that were replicated 4 times. The total AA masses for all treatments were set to 659 mg/L (63% of DMEM) except NEG (0 mg/L). Confluent mammary epithelial cells were exposed to treatment media for 80 min (SD = 7.4). Intracellular concentrations of 17 AA were changed according to F1. The Met and Leu percent of total intracellular AA mass, as an example, varied from 0.58 (PLA) to 6.94 (NEG, +F2) for Met and 0.05 (NEG, -F2) to 4.63 (CGM, +F2) for Leu. Overall, balancing for Lys and Met at a 3:1 ratio increased intracellular concentrations of Lys and Met by 54 and 71%, respectively. Within the mechanistic target of rapamycin (mTOR) pathway, phosphorylation of mTOR (Ser2448), ribosomal protein S6 (Ser235/236), and eukaryotic initiation factor 4E binding protein 1 (Thr37/46) (4EBP1) were increased by all 5 AA profiles compared with the NEG control. We found no differences in phosphorylation state among the 5 AA profiles, indicating lack of sensitivity to various AA profiles. This lack of sensitivity between AA profiles might also be due to assay imprecision or other experimental limitations. Only phosphorylation of 4EBP1 was increased for F2. Phosphorylation of eukaryotic initiation factor 2 α subunit (Ser51) was unaffected by either F1 or F2 factors. Regression analyses indicated that intracellular concentrations of Met, Thr, Ile, and Leu predicted phosphorylation of mTOR-related proteins with adequate precision and accuracy, suggesting that multiple EAA dictate regulation, regardless of AA ratios. Changes in extracellular AA profiles translated to modified intracellular AA profiles, and no single profile uniquely stimulated phosphorylation of the mTOR pathway-related proteins.

Digestible lysine concentrations and amino acid densities influence growth performance and carcass traits in broiler chickens from 14 to 35 days post-hatch

The present study was conducted to evaluate the responses of growth performance and carcass traits in Cobb MV × 500 fast feathering broiler chickens fed maize, soybean meal (SBM) and peanut meal based diets containing different levels of digestible (dig) lysine (Lys). Dietary treatments consisted of 6 concentrations of dig Lys that ranged from 8.4 to 12.9 g/kg in increments of 0.9 g/kg. Two control diets with the medium level of dig Lys concentration of 11.1 g/kg were included where the positive control (PC) diet was a typical commercial diet based on maize and SBM and the negative control (NC) diet replaced part of SBM with100 g/kg peanut meal to allow higher supplementations of feed grade amino acids. Each of the eight experimental diets were offered to either male or female broiler chickens to form 16 experimental treatments and each treatment included 6 floor pens with 12 birds per pen. Broiler chickens were offered experimental diets from 14 to 35 days post-hatch. All diets were balanced with essential amino acids (EAA) by using ideal protein ratio approach and were offered to broiler chickens from 14 to 35 days post-hatch. Predictably, male broiler chickens had better weight gain (WG), feed intake (FI), feed conversion ratio (FCR) and breast meat yield (P < 0.001) than female birds. Sex did not influence yield of wings (P = 0.174) and thighs (P = 0.243). There were no interactions between sex and dietary dig Lys levels on growth performance and linear improvements were observed for live body weight (BW), WG and FCR. By comparing three mathematical models, in female birds, the dig Lys requirement was estimated at 11.5 g/kg for optimal WG and 12.1 g/kg for FCR. In male broiler chickens, the dig Lys requirement was estimated at 10.3 g/kg for optimal WG and 12.2 g/kg for FCR. The results suggested that modern broilers are highly responsive to dietary amino acid (AA) density and regular evaluations of AA requirements are needed because of genetic progress of modern broiler chickens.

The effects of a temporary lysine restriction in newly weaned pigs on growth performance and body composition1.

A serial slaughter study was conducted to determine the effects of a temporary Lys restriction immediately following weaning on growth performance and body composition. One hundred forty-four Yorkshire × Landrace × Duroc pigs (initial BW: 6.9 ± 0.2 kg) were randomly allocated to one of three dietary treatments (six pens per treatment with eight pigs per pen; four barrows and four gilts). For a 3-wk restriction period, pigs were fed diets that were 110% (Control) of the estimated required standardized ileal digestible (SID) Lys for nursery pigs or 20% (Lys20) or 40% (Lys40) below the estimated required SID Lys. Thereafter, all pigs were fed a common grower diet containing 120% of the estimated required SID Lys for 6 wk (recovery period). During the restriction period, ADG and G:F decreased with decreasing dietary Lys concentration (linear; P < 0.01). At the end of the restriction period, BW and whole-body protein concentrations decreased (linear; P < 0.01) and carcass lipid concentrations increased (linear; P < 0.01) with decreasing dietary Lys concentration. During the first 3 wk of the recovery period, ADG and G:F increased (linear; P < 0.05 and P < 0.01, respectively) and whole-body protein concentration at week 3 of the recovery period decreased (linear; P < 0.01) with decreasing dietary Lys concentration. There were no dietary treatment differences in whole-body lipid concentration after 3 wk of the recovery period. During the second half of the recovery period (weeks 7 through 9), there were no differences in ADG or G:F; after week 6, there were no differences in final BW (50.3 ± 0.5 kg) or whole-body protein (16.9 ± 0.2%) or lipid (14.9 ± 0.7%) concentrations. In conclusion, newly weaned pigs previously fed a Lys-limiting diet for 3 wk immediately after weaning achieved full compensatory growth with no differences in BW or body composition after a 6-wk recovery period. Reducing dietary Lys concentration early after weaning is a potential means to reduce the cost of (early) nursery diets without impacting overall growth and carcass composition.

Tear Lactoferrin and Lysozyme as Clinically Relevant Biomarkers of Mucosal Immune Competence.

Tears have attracted interest as a minimally-invasive biological fluid from which to assess biomarkers. Lactoferrin (Lf) and lysozyme (Lys) are abundant in the tear fluid and have antimicrobial properties. Since the eye is a portal for infection transmission, assessment of immune status at the ocular surface may be clinically relevant. Therefore, the aim of this series of studies was to investigate the tear fluid antimicrobial proteins (AMPs) Lf and Lys as biomarkers of mucosal immune status. To be considered biomarkers of interest, we would expect tear AMPs to respond to stressors known to perturb immunity but be robust to confounding variables, and to be lower in participants with heightened risk or incidence of illness. We investigated the relationship between tear AMPs and upper respiratory tract infection (URTI; study 1) as well as the response of tear AMPs to prolonged treadmill exercise (study 2) and dehydration (study 3). Study 1 was a prospective cohort study conducted during the common cold season whereas studies 2 and 3 used repeated-measures crossover designs. In study 1, tear Lys concentration (C) as well as tear AMP secretion rates (SRs) were lower in individuals who reported pathogen-confirmed URTI (n = 9) throughout the observation period than in healthy, pathogen-free controls (n = 17; Lys-C, P = 0.002, d = 0.85; Lys-SR, P < 0.001, d = 1.00; Lf-SR, P = 0.018, d = 0.66). Tear AMP secretion rates were also lower in contact lens wearers. In study 2, tear AMP SRs were 42–49% lower at 30 min−1 h post-exercise vs. pre-exercise (P < 0.001, d = 0.80–0.93). Finally, in study 3, tear AMPs were not influenced by dehydration, although tear AMP concentrations (but not secretion rates) displayed diurnal variation. We conclude that Lf and Lys have potential as biomarkers of mucosal immune competence; in particular, whether these markers are lower in infection-prone individuals warrants further investigation.

Evidence by GC-MS that lysine is an arginase-catalyzed metabolite of homoarginine in vitro and in vivo in humans

l-Homoarginine (hArg) is biosynthesized from l-arginine (Arg) and l-lysine (Lys) by arginine:glycine amidinotransferase (AGAT). AGAT also catalyzes the formation of guanidinoacetate (GAA) from Arg and glycine (Gly). GAA is converted to creatine (N-methyl guanidinoacetate) by guanidinoacetate N-methyl-transferase (GAMT). Low circulating and excretory concentrations of hArg are associated with worse cardiovascular outcome and mortality. hArg is a poor substrate of nitric oxide synthase (NOS) and a weak inhibitor of arginase. The metabolism of hArg in humans is little investigated. Previously, we found that orally administered hArg (125 mg/day) increased the plasma concentration of hArg, but not of Arg, the substrate of NOS, in healthy subjects. We newly analyzed the plasma samples collected in that study for Lys and other amino acids. Repeated measures ANOVA revealed statistically significant differences between the groups (P = 0.008) with respect to plasma Lys concentration which increased by about 8% after a 4-week hArg supplementation. In vitro, recombinant human arginase and bovine liver arginase I were demonstrated by a specific and sensitive stable-isotope GC-MS assay to hydrolyze hArg to Lys. Our results suggest that Lys is a metabolite of hArg produced by the hydrolytic activity of arginase. Arginase may play a key role in hArg homeostasis in humans.

Box-Behnken optimisation of growth performance, plasma metabolites and carcass traits as influenced by dietary energy, amino acid and starch to lipid ratios in broiler chickens.

A Box-Behnken designed study was completed to predict growth performance, carcass characteristics and plasma hormone and metabolite levels as influenced by dietary energy, amino acid densities and starch to lipid ratios in male broiler chickens. The design comprised three dietary energy densities (11.25, 12.375 and 13.5 MJ/kg), three digestible lysine concentrations (9.2, 10.65 and 12.1 g/kg) and three starch to lipid ratios (4.5, 12.25 and 20.0) in broiler diets based on maize and soybean meal. Each of thirteen dietary treatments was offered to 10 replicates of 15 birds per replicate floor pen or a total of 1,950 Ross 308 male broiler chickens from 21 to 35 days post-hatch. Increasing dietary energy decreased feed intake with a quadratic relationship between feed intake and dietary standardised ileal digestible (SID) Lys concentrations, where increasing SID Lys initially increased and then depressed feed intake. Increasing dietary amino acid density increased body weight gain and carcass weight; however, dietary energy did not influence body weight gain, carcass and breast meat weight. Feed efficiency was positively influenced by energy and amino acid densities but negatively influenced by starch to lipid ratios and energy and amino acids had more pronounced impacts than starch to lipid ratios. This study indicated that both energy and amino acid densities regulate feed intakes in broiler chickens. Body weight gain of modern broiler chickens is more responsive to amino acid densities; nevertheless, dietary energy density continues to play an important role in protein utilisation, as reflected in significantly reduced plasma uric acid levels.

Assessing bioavailability of ruminally protected methionine and lysine prototypes

Met and Lys are essential AA that can limit lactational performance in dairy cattle fed protein-sufficient diets. Thus, there is industry demand for ruminally protected (RP) sources of Met and Lys. One method of providing ruminal protection for Met and Lys is lipid encapsulation. The objective of this work was to assess 3 lipid-encapsulated Met prototypes (P1, P2, and P3) and 1 Lys prototype (P4) to determine ruminal protection, small intestine absorption (experiment 1), and animal production responses (experiment 2). Ruminal protection was estimated from 8-h in situ retention during ruminal incubation and intestinal absorption from plasma appearance after an abomasal bolus of the in situ retentate. Blood samples were collected over time to determine plasma Met and Lys concentration responses compared with unprotected Lys and Met infused abomasally. The prototypes were not exposed to the total diet or subjected to typical feed handling methods before evaluation. The bioavailability of P1, P2, and P3 Met prototypes was found to be 14, 21, and 18% of the initial AA material, respectively. The RP-Lys prototype had a bioavailability of 45%. To evaluate production responses, 20 Holstein cows were randomly assigned to 2 trials (n = 10 each) in a replicated Latin square design with 14-d periods. The base diet was predicted to be deficient in metabolizable Met (-14.8 g/d) and Lys (-16.1 g/d) per the Cornell Net Carbohydrate and Protein System (version 6.55). In the Met trial, the base diet was supplemented with RP-Lys to meet Lys requirements, and treatments were as follows: no added RP-Met (NCM), NCM plus Smartamine M (SM; Adisseo, Alpharetta, GA), and NCM plus P1, P2, or P3 at 148% of the Met content of SM. In the Lys trial, the base diet was supplemented with RP-Met to meet the Met requirement, and treatments were as follows: no added Lys (NCL), NCL plus AjiProL (AL; Ajinomoto Heartland Inc., Chicago, IL), and NCL plus P4 at 55, 78, or 102% of the reported absorbed Lys in AL. All products were top dressed on the diet without prior mixing or extended exposure to the rest of the diet. Milk protein concentration significantly increased when diets were supplemented with P2, P3, or SM (3.12, 3.12, and 3.11%, respectively) compared with NCM (3.02%). Only P1 (3.04%) was significantly lower than SM. Prototype P2 had the greatest numerical milk protein output response among the 3 RP-Met prototypes, suggesting that it may have had the greatest efficacy when supplemented into these rations. There was a numerical milk protein concentration response to AL and a linear increase in milk protein concentration for P4. The P4 and AL treatments resulted in comparable milk protein production regardless of P4 dose.

The effect of Arginine, Lysine and Histidine in the myosin secondary structure by circular dichroism and Raman spectroscopy

ABSTRACTAmino acids containing L-Arginine (Arg), L-Lysine (Lys) and L-Histidine (His) are recognized to increase the solubility of myosin at low ionic strength, but there is a difference in the researchers’ views on their effects on the secondary structure of myosin. To elucidate which method is relatively appropriate for the determination of secondary structures of myosin added with Arg, Lys or His, Circular dichroism (CD) spectra and Raman spectroscopy were carried out respectively. Raman spectroscopy was more suitable than CD method for determination of the secondary structures of myosin added with His, while CD spectra was more accurate for the myosin samples added with Lys and the adequate concentration of Lys should be under 10 mM. Both CD and Raman methods could be applied to myosin added with Arg, however, the latter was recommended and the adequate additive content of Arg in CD method should be less than 10 mM.

Efficiently activated ε-poly-L-lysine production by multiple antibiotic-resistance mutations and acidic pH shock optimization in Streptomyces albulus.

ε‐Poly‐L‐lysine (ε‐PL) is a food additive produced by Streptomyces and is widely used in many countries. Working with Streptomyces albulus FEEL‐1, we established a method to activate ε‐PL synthesis by successive introduction of multiple antibiotic‐resistance mutations. Sextuple mutant R6 was finally developed by screening for resistance to six antibiotics and produced 4.41 g/L of ε‐PL in a shake flask, which is 2.75‐fold higher than the level produced by the parent strain. In a previous study, we constructed a double‐resistance mutant, SG‐31, with high ε‐PL production of 3.83 g/L and 59.50 g/L in a shake flask and 5‐L bioreactor, respectively. However, we found that R6 did not show obvious advantages in fed‐batch fermentation when compared with SG‐31. For further activation of ε‐PL synthesis ability, we optimized the fermentation process by using an effective acidic pH shock strategy, by which R6 synthetized 70.3 g/L of ε‐PL, 2.79‐fold and 1.18‐fold greater than that synthetized by FEEL‐1 and SG‐31, respectively. To the best of our knowledge, this is the highest reported ε‐PL production to date. This ε‐PL overproduction may be due to the result of R99P and Q856H mutations in ribosomal protein S12 and RNA polymerase, respectively, which may be responsible for the increased transcription of the ε‐poly‐lysine synthetase gene (pls) and key enzyme activities in the Lys synthesis metabolic pathway. Consequently, ε‐PL synthetase activity, intracellular ATP, and Lys concentrations were improved and directly contributed to ε‐PL overproduction. This study combined ribosome engineering, high‐throughput screening, and targeted strategy optimization to accelerate ε‐PL production and probe the fermentation characteristics of hyperyield mutants. The information presented here may be useful for other natural products produced by Streptomyces.

Estimated energy and nutrient composition of different sources of food waste and their potential for use in sustainable swine feeding programs.

About 40% of the total food produced in the United States is wasted throughout the supply chain. The objective of this study was to determine the energy and nutrient content and variability of food waste sources generated at different stages within the food supply chain in the Minneapolis–St. Paul, MN, metropolitan area, and their potential for use in swine diets. A total of four waste sources were selected: supermarket (SM; retail to consumer), university residential dining hall (RH; consumer to postconsumer), a city waste transfer station (TS; postconsumer to municipal waste disposal), and household source-separated organic recycling program (SSO; postconsumer to municipal waste). Samples were collected (SM: n = 22; RH: n = 60; TS: n = 27; SSO: n =12) and analyzed for GE, proximate analyses, minerals, amino acids, and fatty acid concentrations along with lipid peroxidation indicators including peroxide value (PV) and thiobarbituric reactive substances (TBARS). Data were analyzed using a general linear model that included food waste source as the main factor, and least squared means with adjustment were used for multiple comparisons. Samples of SM food waste contained the greatest (P < 0.05) concentration of GE (5,909 kcal/kg) compared with RH, TS, and SSO sources. Calculated NE of SM (3,740 kcal/kg) was also the greatest compared with the three other food waste sources. Food waste from SM, RH, and SSO, but not TS, had greater (P < 0.05) calculated NE than published values for corn and soybean meal. Concentrations of Lys (1.82%), Met (0.53%), Thr (1.07%), and Trp (0.27) content were greater in SM than in RH, TS, and SSO, but these concentrations were less than published values for soybean meal. There were no differences (P > 0.05) in the phosphorus content of samples among food waste sources (0.30% to 0.64%). PV and TBARS were greatest (P < 0.05) in the SSO samples (PV = 82.4 meq/kg oil; TBARS = 2.44 mg malondialdehyde (MDA) eq/g oil) compared with the other three food waste sources. Although the concentrations of nutrients and calculated energy values of the food waste sources were moderately high compared with corn and soybean meal, their composition was more variable (i.e., greater SD of means). Food waste generated upstream (SM) in the food supply chain appears to have greater nutritional value than postconsumer food waste (RH, TS, and SSO), but all sources appear suitable for use in commercial swine diets provided that ME, NE, and nutrient digestibility values are well characterized.

Effect of ractopamine on digestible-lysine requirement for finishing barrows under thermoneutral conditions

The purpose of the present study was to determine the digestible-lysine (Lys) requirement in diets supplemented or not with ractopamine (RAC), so as to maximise performance and carcass characteristics for finishing barrows reared at thermoneutral conditions. In total, 128 high-lean commercial hybrid barrows with an initial bodyweight of 78.2 ± 4.3 kg were distributed in a completely randomised block design, in a 4 × 2 factorial arrangement (four Lys concentrations: 0.730%, 0.830%, 0.930% and 1.030% and two RAC inclusions: 0 and 10 mg/kg), with two animals per pen and eight pens per treatment. Lys concentrations tested significantly (P &amp;lt; 0.05) affected digestible-lysine intake, and the absolute (g) and relative (%) lean meat content, regardless of RAC inclusion. When diets were RAC-supplemented, feed conversion rate of animals was improved and, in addition, final bodyweight, daily bodyweight gain, loin eye area, loin eye depth, and absolute (g) and relative (%) carcass lean meat content increased (P &amp;lt; 0.05). RAC inclusion also decreased (P &amp;lt; 0.05) backfat thickness and the colour values a* and b* of finishing pigs. Therefore, on the basis of our results, it can be inferred that the best performance and carcass characteristics for finishing barrows reared at thermoneutral conditions were obtained at 1.030% and 0.730% Lys respectively, corresponding to an estimated daily Lys intake of 34.3 g and 24.2 g, independently of the diet RAC concentration.

Effect of standardized ileal digestible lysine on growth and subsequent performance of weanling pigs.

A total of 320 weanling pigs (all barrows, initially 5.71 kg BW; Line 1050, PIC Hendersonville, TN) were used to determine whether the Lys level fed during one phase of the nursery influences the response to Lys during subsequent phases. Our hypothesis was that feeding decreasing dietary Lys concentration in early phases, but feeding adequate concentrations in later phases might result in similar pig growth as those fed a more conventional approach with step-wise decreases in dietary Lys as pigs become heavier. Eight dietary regimens were used in a split-plot design. There were three dietary phases, and within phase, a high or low standardized ileal digestible (SID) Lys diet was fed. Pigs were fed either 1.35% or 1.55% SID Lys during phase 1 (days 0 to 7), 1.15% or 1.35% SID Lys in phase 2 (days 7 to 21), and 1.05 or 1.25% SID Lys during phase 3 (days 21 to 35). The low dietary Lys concentrations were achieved by reducing both crystalline Lys and intact protein sources from the high Lys diets. From days 0 to 7, feeding high SID Lys improved (P < 0.01) G:F, but no evidence for differences in ADG or ADFI were observed. Similarly, from days 7 to 21, there were no evidence for differences in ADG or ADFI among pigs fed the two Lys levels, but those fed high SID Lys had improved (P < 0.03) G:F. From days 21 to 35, pigs fed the high Lys diet had increased (P < 0.01) ADG and G:F compared with those fed low SID Lys, but there were no effects on ADFI. For the overall trial (days 0 to 35), there were no dietary interactions among phases, indicating that the Lys level fed in each phase did not influence the response to Lys in subsequent phases. Thus, pigs fed the high Lys level during phase 3, regardless of previous Lys levels in phases 1 and 2, had greater (P < 0.05) overall ADG and G:F compared with other treatment groups. In conclusion, relatively low dietary Lys concentrations can be fed in the early nursery phases (approximately 6 to 12 kg) without any negative impact on overall growth performance provided that adequate Lys levels are fed thereafter (12 to 20 kg).

A comparison of milk protein, fat, lactose, total solids and amino acid profiles of three different buffalo breeds in Guangxi, China

This study compared the milk protein, fat, lactose, total solids and amino acids (AA) profiles of Murrah buffalo, Nili-Ravi buffalo and crossbreed buffalo. Buffalo milk samples of Murrah (n = 25), Nili-Ravi (n = 20) and Triple-crossbred (n = 23) were collected and milk protein, fat, lactose, total solids and 17 AA profiles were determined. Milk protein and total solids contents of Murrah buffalo were higher than those of Nili-Ravi buffalo (p < .05). The average milk protein, fat, lactose and total solids contents of all buffalo samples were 4.76, 7.31, 5.19 and 18.40 g/100 g of milk, respectively. The AA composition of all three kinds of milk samples was rich in glutamic acid (0.89–0.96 g/100 g of milk) and poor in cysteine (0.01–0.02 g/100 g of milk). Leucine, lysine, valine and isoleucine were the most prevalent essential amino acids; while glutamic acid, proline, aspartic acid and serine were the most prevalent non-essential amino acids in the three kinds of milk samples. Significant differences (p < .05) were observed for concentrations of Lys, Ile, Leu, Phe, Cys and His, while the other 11 amino acids had no significant differences (p > .05). Conclusively, the milk of Murrah buffalo, Nili-Ravi buffalo and crossbreed buffalo was all rich in glutamic acid while poor in cysteine contents. After crossbreeding, the milk production of Triple-crossbred buffalo was higher than the local Chinese buffalo and its milk composition was in between those of Murrah and Nili-Ravi.

Blocking amino acid transporter OsAAP3 improves grain yield by promoting outgrowth buds and increasing tiller number in rice.

SummaryAmino acid transporters (AATs) play indispensable roles in nutrient allocation during plant development. In this study, we demonstrated that inhibiting expression of the rice amino acid transporter OsAAP3 increased grain yield due to a formation of larger numbers of tillers as a result of increased bud outgrowth. Elevated expression of OsAAP3 in transgenic plants resulted in significantly higher amino acid concentrations of Lys, Arg, His, Asp, Ala, Gln, Gly, Thr and Tyr, and inhibited bud outgrowth and rice tillering. However, RNAi of OsAAP3 decreased significantly Arg, Lys, Asp and Thr concentrations to a small extent, and thus promoted bud outgrowth, increased significantly tiller numbers and effective panicle numbers per plant, and further enhanced significantly grain yield and nitrogen use efficiency (NUE). The promoter sequences of OsAAP3 showed some divergence between Japonica and Indica rice, and expression of the gene was higher in Japonica, which produced fewer tillers than Indica. We generated knockout lines of OsAAP3 on Japonica ZH11 and KY131 using CRISPR technology and found that grain yield could be increased significantly. These results suggest that manipulation of OsAAP3 expression could be used to increase grain yield in rice.

Creating magnetic ionic liquid-molecularly imprinted polymers for selective extraction of lysozyme.

A novel magnetic (Fe3O4) surface molecularly imprinted polymer (MIP) based on ionic liquid (IL) (Fe3O4@VTEO@IL-MIPs) was prepared for the selective extraction of lysozyme (Lys). As the functional monomer of the MIPs, an imidazolium-based IL with vinyl groups was prepared. It can provide multiple interactions with template molecules. The amount of IL was optimized (200 mg). Fourier transform infrared spectrometry (FT-IR), transmission electron microscopy (TEM), dynamic light scattering (DLS), thermogravimetric analysis (TGA) and a vibrating sample magnetometer (VSM) were used to characterize the MIP. The results indicate the successful formation of an imprinting polymer layer. The concentration of Lys in the supernatant was determined by UV-vis spectrophotometry at a wavelength of 280 nm. The maximum adsorption capability of the MIP is 213.7 mg g−1 and the imprinting factor (IF) is 2.02. It took 2.5 h for the MIP to attain adsorption equilibrium. The structure of the protein was evaluated using circular dichroism (CD) spectra and UV-visible spectra. The adsorption performance was further investigated in detail by selective adsorption experiments, competitive rebinding tests, and reusability and stability experiments. Furthermore, it was utilized to separate the template protein from a mixture of proteins and real samples successfully because of the high adsorption capacity for Lys.

Salicylicylphenylacetylene fluorophore mixed with graphene oxide for selective lysozyme detection

A highly selective method of lysozyme detection based on salicylicylphenylacetylene (1) fluorophores and graphene oxide (GO) was successfully developed. The fluorescent emission of 1 was decreased fivefold in the addition of GO (0.6 Abs) due to π-π and hydrophobic interaction between 1 and GO graphitic plane. In the presence of lysozyme (Lys), the non-emissive 1-GO became highly fluorescent as a result of strong interaction of 1 with the cationic Lys preventing the complexation between 1 and GO. The linear dynamic response of fluorescence intensity was observed in the Lys concentration range of 0–80µg/mL with the limit of detection of 65ng/mL. This label free technique was developed for quantitative determination of Lys in egg white sample with an analytical accuracy of less than 10% error.

Varying the ratio of Lys:Met while maintaining the ratios of Thr:Phe, Lys:Thr, Lys:His, and Lys:Val alters mammary cellular metabolites, mammalian target of rapamycin signaling, and gene transcription

Amino acids are not only precursors for but also signaling molecules regulating protein synthesis. Regulation of protein synthesis via AA occurs at least in part by alterations in the phosphorylation status of mammalian target of rapamycin (mTOR) pathway proteins. Although the ideal profile of Lys:Met to promote milk protein synthesis during established lactation in dairy cows has been proposed to be 3:1, aside from being the most-limiting AA for milk protein synthesis, the role of Met in other key biologic pathways such as methylation is not well characterized in the bovine. The objective of this study was to determine the influence of increasing supplemental Met, based on the ideal 3:1 ratio of Lys to Met, on intracellular metabolism related to protein synthesis and mTOR pathway phosphorylation status. MAC-T cells, an immortalized bovine mammary epithelial cell line, were incubated (n = 5 replicates/treatment) for 12 h with 3 incremental doses of Met while holding Lys concentration constant to achieve the following: Lys:Met 2.9:1 (ideal AA ratio; IPAA), Lys:Met 2.5:1 (LM2.5), and Lys:Met 2.0:1 (LM2.0). The ratios of Thr:Phe (1.05:1), Lys:Thr (1.8:1), Lys:His (2.38:1), and Lys:Val (1.23:1) were the same across the 3 treatments. Applying gas chromatography-mass spectrometry metabolomics revealed distinct clusters of differentially concentrated metabolites in response to Lys:Met. Lower Phe, branched-chain AA, and putrescine concentrations were observed with LM2.5 compared with IPAA. Apart from greater intracellular Met concentrations, further elevations in Met level (LM2.0) led to greater intracellular concentrations of nonessential AA (Pro, Glu, Gln, and Gly) compared with IPAA and greater essential AA (EAA; Met, Ile, and Leu) and nonessential AA (Pro, Gly, Ala, Gln, and Glu) compared with LM2.5. However, compared with IPAA, mRNA expression of β-casein and AA transporters (SLC7A5, SLC36A1, SLC38A2, SLC38A9, and SLC43A1) and mTOR phosphorylation were lower in response to LM2.5 and LM2.0. Overall, the results of this study provide evidence that increasing Met while Lys and the ratios of Phe, Thr, His, and Val relative to Lys were held constant could increase the concentration and utilization of intracellular EAA, in particular branched-chain AA, potentially through improving the activity of AA transporters partly controlled by mTOR signaling. Because EAA likely are metabolized by other tissues upon absorption, a question for future in vivo studies is whether formulating diets for optimal ratios of EAA in the metabolizable protein is sufficient to provide the desired levels of these AA to the mammary cells.

Modeling the effects of standardized ileal digestible valine to lysine ratio on growth performance of nursery pigs.

Two experiments evaluated the effects of increasing Lys and Val on growth performance of nursery pigs. In Exp. 1,300 nursery pigs (PIC 327 × 1,050, initially 6.7 ± 1.4 kg BW) were randomly allotted to 1 of 6 diets containing 1.10, 1.20, 1.30, 1.40, 1.50, or 1.60% standardized ileal digestible (SID) Lys, with 10 pens per dietary treatment and 5 pigs per pen. Linear and nonlinear mixed models were fitted to estimate dose responses. From d 0 to 14, and for the overall 28 d period, ADG and G:F increased (linear, P < 0.001) as SID Lys increased, with no evidence of differences in ADFI. Dose response modeling indicated the SID Lys requirement for ADG and G:F was at 1.45% using a broken line linear (BLL) and greater than 1.60% using a quadratic polynomial (QP) model. In Exp. 2, 280 nursery pigs (PIC 327 × 1,050, initially 6.5 ± 1.3 kg BW) were allotted to 1 of 7 diets containing SID Val:Lys ratios of 50, 57, 63, 68, 73, 78, or 85%. The dietary SID Lys concentration 1.24% SID Lys which was below the estimated requirement from Exp. 1 and ensured the Val:Lys ratio was not underestimated. From d 0 to 14, ADG, ADFI, and G:F increased (quadratic, P < 0.039) with increasing SID Val:Lys. For ADG, the best fitting model was a BLL, with a breakpoint estimate of 62.9% SID Val:Lys [52.2, 73.7] ratio while for G:F the best fit model was a quadratic polynomial with a maximum G:F at 71.7% SID Val:Lys (95%CI:[58, > 85]). Average daily feed intake was also modeled with a quadratic polynomial and maximized at 73.7% Val:Lys (95% CI: [61, > 85]). In conclusion, the Val requirement ranged from approximately 63 to 74% of Lys depending on the response criteria modeled.

Effects of combined α-galactosidase and xylanase supplementation on nutrient digestibility and growth performance in growing pigs

ABSTRACTTwo experiments were conducted to investigate the effect of combined supplementation of α-galactosidase and xylanase on nutrient digestibility and growth performance in growing pigs. Experiment 1 had a 2 × 2 Latin square design, where eight barrows (45.0 ± 0.52 kg body weight [BW]) were fitted with a simple T-cannula in the distal ileum and received a basal diet without or with supplementation of α-galactosidase (12 U/kg diet) and xylanase (15 AXC/kg diet) within two periods of 10 d. The apparent ileal digestibility (AID) and apparent total tract digestibility of nutrients, pH, viscosity of digesta and digestive enzyme activities were assessed. In Experiment 2, a total of 432 growing pigs (initial BW 44.7 ± 0.66 kg) were allocated to four treatments. Diets were based on corn and soybean meal and had a normal or reduced nutrient level (reduced by 0.42 kJ digestible energy [DE] per kg and 0.8% crude protein). Both diets were offered without or with supplementation of α-galactosidase and xylanase. The growth performance was assessed within a 43-d feeding period, where at the end, biochemical serum indices were estimated. In Experiment 1, the enzyme-supplemented diet had a greater contents of DE and DE/gross energy ratio (p < 0.05), and a higher AID of Arg, raffinose, stachyose and arabinoxylan (p < 0.05). In Experiment 2, the low nutrient level caused lower daily gain (p < 0.05), which was partially compensated by enzyme addition. Enzyme addition also increased the serum concentration of Lys (p < 0.05). Moreover, it appears that the tested enzyme supplementation could increase dietary DE, serum total amino acid concentrations and decrease serum urea nitrogen.