Effects of dietary amino acid supplementation on measures of whole‐body and muscle protein metabolism in aged horses

The objective of this study was to examine markers of whole‐body and muscle protein metabolism in aged horses fed a diet typical for North American aged horses, supplemented with amino acids. In a replicated Latin square design, six aged horses (20 ± 1.1 years) were studied while receiving each of three isocaloric, isonitrogenous diets, a control treatment concentrate (CON; 100 mg/kg^?1 BW day^?1 lysine, 84 mg kg^?1 day^?1 threonine, 51 mg kg^?1 day^?1 methionine), LYS/THR (134 mg kg^?1 BW day^?1 lysine, 110 mg kg^?1 BW day^?1 threonine, 52 mg kg^?1 BW day^?1 methionine) and LYS/THR/MET (132 mg kg^?1 BW day^?1 lysine, 112 mg kg^?1 BW day^?1 threonine, 62 mg kg^?1 BW day^?1 methionine). In each 15‐days period, urine and faeces were collected for assessment of nitrogen balance. Blood samples were collected before and after feeding for analysis of plasma urea nitrogen (PUN), glucose, insulin and plasma amino acid concentrations. Skeletal muscle samples were collected for measurement of proteins associated with muscle protein synthesis and degradation, and horses underwent stable isotope infusion procedures for comparison of differences in whole‐body rates of protein synthesis and degradation. There was no effect of treatment on relative abundance of proteins involved in protein synthesis, nitrogen retention or phenylalanine kinetics. PUN concentrations tended to be higher for LYS/THR (p = 0.054) and were higher for LYS/THR/MET (p = 0.0056) than for CON. Atrogin‐1 abundance tended to be higher in the post‐absorptive state for the CON treatment (p = 0.07), indicating that amino acid supplementation resulted in less muscle protein degradation when horses were in the post‐absorptive state. However, lack of differences in nitrogen retention and phenylalanine kinetics indicated that whole‐body protein metabolism was not improved, and higher PUN concentrations in the supplemented diets suggest that the supplemented amino acids may have been catabolized. Amino acid availability was not limiting protein synthesis in the sedentary aged horses in this study when fed the CON diet.     

Postprandial oxidative losses of free and protein‐bound amino acids in the diet: interactions and adaptation

Postprandial oxidation of dietary free amino acids or egg white protein was studied using the [¹³CO₂] breath test in rats, as well as in humans. Thirty‐eight male rats were assigned to four dietary test groups. Two diets only differed in their protein fraction. Diet I contained 21% egg white protein. For the breath test egg white protein, intrinsically labelled with [1‐¹³C]‐leucine, was used as a substrate. Diet II contained the same amino acids as diet I, though not as egg white protein but in free form. Free [1‐¹³C]‐leucine was used to label this diet. In addition, two 1:1 mixtures of both diets were used. During the breath test either the free amino acid or the protein fraction was labelled as in diets I or II. The animals were breath‐tested following short‐term (day 5) and long‐term adaptation (day 20) to their experimental diet. For all diets, including the mixed diets, postprandial oxidative losses on day 5 were significantly higher for the free leucine compared with the protein‐derived leucine. Differences between free and protein‐derived leucine oxidation had, however, largely disappeared on day 20. The human subjects were breath‐tested without any adaptation period to the diets. The oxidative losses of free leucine were also higher than those of protein‐derived leucine. None of the studies showed any indication for an interaction between the oxidation of protein‐derived amino acids and free amino acids. It is concluded that free and protein‐derived amino acids in the diet are mainly metabolized independently.     

Effects of phenylalanine and threonine oligopeptides on milk protein synthesis in cultured bovine mammary epithelial cells

This study was conducted to investigate the effects of phenylalanine (Phe) and threonine (Thr) oligopeptides on α_s1 casein gene expression and milk protein synthesis in bovine mammary epithelial cells. Primary mammary epithelial cells were obtained from Holstein dairy cows and incubated in Dulbecco's modified Eagle's medium‐F12 medium (DMEM/F12) containing lactogenic hormones (prolactin and glucocorticoids). Free Phe (117 μg/ml) was substituted partly with peptide‐bound Phe (phenylalanylphenylalanine, phenylalanyl threonine, threonyl‐phenylalanyl‐phenylalanine) in the experimental media. After incubation with experimental medium, cells were collected for gene expression analysis and medium was collected for milk protein or amino acid determination. The results showed that peptide‐bound Phe at 10% (11.7 μg/ml) significantly enhanced α_s1 casein gene expression and milk protein synthesis as compared with equivalent amount of free Phe. When 10% Phe was replaced by phenylalanylphenylalanine, the disappearance of most essential amino acids increased significantly, and gene expression of peptide transporter 2 and some amino acid transporters was significantly enhanced. These results indicate that the Phe and Thr oligopeptides are important for milk protein synthesis, and peptide‐bound amino acids could be utilised more efficiently in milk protein synthesis than the equivalent amount of free amino acids.     

Effect of Chinese herbal medicine on kinetics of plasma phenylalanine,tyrosine and whole body protein synthesis in sheep

The aim of this study was to evaluate the effect of feeding decoction of a traditional nourishing Chinese herbal medicine formula on rates of plasma phenylalanine and tyrosine turnover and whole body protein synthesis in sheep. Ruminal fermentation characteristics and blood metabolites were also determined. Six sheep were subjected to either mixed hay (MH‐diet, as control) or MH‐diet supplemented with 2% of Chinese herbal medicine (mixture of Astragalus root, Angelica root, and Atractylodes rhizome; CHM‐diet) in a crossover design for each of 3‐week period. The isotope dilution of [²H₅]phenylalanine and [²H₂]tyrosine was performed as a primed‐continuous infusion to measure plasma phenylalanine and tyrosine kinetics. Concentrations of total volatile fatty acid, acetate, and propionate in the rumen tended to be higher (p < 0.10), and the pH value was lower (p = 0.04) for the CHM‐diet than the MH‐diet. Turnover rates of plasma phenylalanine and tyrosine tended to be higher (p < 0.10) for the CHM‐diet than the MH‐diet. Furthermore, whole body protein synthesis was greater (p = 0.04) for the CHM‐diet compared with the MH‐diet. The Chinese herbal medicine improved rumen fermentation and enhanced protein metabolism in sheep. Hence, it is suggested that the decoction of Chinese herbal medicine formula could be considered as a potential feed additive for ruminant production.     

Apparent and standardized ileal digestibility of amino acids in diverse barley cultivars fed to growing pigs

The objectives of this study were to determine the chemical composition, ileal amino acid (AA) digestibility of barley varieties and to develop equations to predict the AA digestibility based on the chemical characteristics of individual barley. A wide selection of nine barley varieties in the main growing regions of China was used in the experiment. Ten crossbred pigs (30.9 ± 1.8 kg) were allotted to a 6 × 10 Youden Square. The treatments included a nitrogen‐free diet and nine barley test diets. Considerable variation was observed in the content of neutral detergent fiber, acid detergent fiber, crude protein (CP) and all AAs among the barley samples. The standardized ileal digestibility (SID) of indispensable AAs with the exception of arginine, phenylalanine and threonine were different (P < 0.05) among the nine barley sources tested. With R² values exceeding 0.50, significant linear regression equations for CP and threonine were obtained. In conclusion, significant differences existed in the ileal digestibility of CP and AAs among barley varieties. It is difficult to predict the SID of most of the amino acids in barely based upon chemical characteristics.     

A Preliminary Study of 13C-Phenylalanine and 13C-Dipeptide Breath Tests in Horses

This study aimed to establish a standard dose and sample collection time for ¹³C phenylalanine and ¹³C-Dipeptide breath test in horses. To evaluate dose-dependent effects, healthy horses received 2.5 mg/kg, 5 mg/kg, and 10 mg/kg ¹³C phenylalanine dissolved in 1 ml/kg distilled water and 1.25 mg/kg, 2.5 mg/kg, and 5 mg/kg ¹³C dipeptide dissolved in 2 ml/ kg distilled water. Tmax was observed during the sample collection time. For ¹³C phenylalanine, the standard deviation of Cmax at 5 mg/kg was lower than that of 10 mg/kg. For ¹³C dipeptide, the standard deviation of Tmax was the lowest at 5 mg/kg. This study revealed that an optimal dose for breath tests with ¹³C phenylalanine and ¹³C dipeptide may be 5 mg/kg in horses.     

Influence of an n‐3 long‐chain polyunsaturated fatty acid‐enriched diet on experimentally induced synovitis in horses

Dietary n‐3 long‐chain polyunsaturated fatty acid (LCPUFA) supplementation has previously been shown to modify joint‐related inflammation in several species, although information in the horse is lacking. We investigated whether dietary supplementation with n‐3 LCPUFA would modify experimentally induced synovitis in horses. Twelve, skeletally mature, non‐pregnant mares were randomly assigned to either a control diet (CONT) or an n‐3 long‐chain fatty acid‐enriched treatment diet (N3FA) containing 40 g/day of n‐3 LCPUFA for 91 days. Blood samples taken on days 0, 30, 60 and 90, and synovial fluid collected on days 0 and 90 were processed for lipid composition. On day 91, joint inflammation was stimulated using an intra‐articular (IA) injection of 100 ng of recombinant equine IL‐1beta (reIL‐1β). Synovial fluid samples taken at post‐injection hours (PIH) 0, 4, 8 and 24 were analysed for prostaglandin E₂ (PGE₂), matrix metalloproteinase (MMP) activity and routine cytology. Synovium and articular cartilage samples collected at PIH 8 were analysed for gene expression of MMP 1 and MMP 13, interleukin‐1beta (IL‐1β), cyclooxygenase 2 (COX‐2), tumour necrosis factor‐alpha and the aggrecanases, a disintegrin and metalloprotease with thrombospondin motifs (ADAMTS)‐4 and ADAMTS‐5. A 90‐day feeding period of n‐3 LCPUFA increased serum phospholipid and synovial fluid lipid compositions of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) compared to CONT horses. The reIL‐1β injection caused an inflammatory response; however, there was no effect of dietary treatment on synovial fluid PGE2 content and MMP activity. Synovial tissue collected from N3FA horses exhibited lower expression of ADAMTS‐4 compared to CONT horses. Despite the presence of EPA and DHA in the synovial fluid of N3FA horses, dietary n‐3 LCPUFA supplementation did not modify synovial fluid biomarkers compared to CONT horses; however, the lower ADAMTS‐4 mRNA expression in N3FA synovium warrants further investigation of n‐3 LCPUFA as a joint therapy.     

Establishing an essential amino acid profile for maintenance in poultry using deletion method

This study aimed to estimate the essential amino acid profile and the ideal ratio for the maintenance of poultry by deletion method. A nitrogen balance (NB) trial was conducted using 198 adult roosters, housed individually in metabolic cages. The treatments were 33 purified diets being 11 diets with an amino acid mixture providing high protein intake of 500 mg N/BW_kg^0.75 per day, 11 diets providing medium protein intake of 250 mg N/BW_kg^0.75 per day (in each diet, one amino acid tested was diluted 50%) and 11 diets providing low protein intake of 125 mg N/BW_kg^0.75 per day (made by omitting the amino acid tested). Each treatment had six replicates. After 48 h of fasting receiving water plus sucrose, the roosters were fed 40 g of the diets by tube once a day for 3 days. The excreta were collected within 72 h after the first feeding. The diets and excreta were analysed for nitrogen content. For each amino acid studied, a linear regression was fitted by NB and amino acid intake (AAI). The maintenance requirements were estimated as the AAI to maintain the NB equal to zero. The daily amino acid requirements for maintenance were estimated to be Lys 11, Met 29, Thr 23, Trp 5, Arg 50, Val 29, His 6, Gly 54, Phe 49, Leu 78 and Ile 21 mg/BW_kg^0.75 per day. Therefore, the amino acid ratio for maintenance was concluded to be Lys 100, Met 276, Thr 220, Trp 48, Arg 467, Val 275, His 60, Gly 511, Phe 467, Leu 735 and Ile 198% independent of the scale. The essential amino acid profile and the ideal ratio for the maintenance of poultry estimated in this study contributed to improve the factorial model for estimating essential amino acid requirements for poultry.     

Oral and intravenous l‐[1‐13C]phenylalanine delivery measure similar rates of elimination when gastric emptying and splanchnic extraction are accounted for in adult mixed hounds1–4

There are few reported estimates of amino acid (AA) kinetics in adult mammals and none exist in adult dogs. The study objectives were to evaluate the use of oral isotope delivery in contrast to the more commonly used intravenous (IV) delivery to estimate AA kinetics in adult dogs and to estimate splanchnic extraction and gastric emptying using a commonly accepted mathematical model. Dogs received 25 × 1/2‐hourly meals (13 g/kg BW/day) and either an oral or IV bolus of l‐ [1‐¹³C]Phe (12 mg/kg BW). Blood samples were taken immediately before each feeding. Concentrations of plasma Phe were measured using liquid chromatography‐tandem mass spectrometry. There were no differences in baseline plasma Phe concentrations (34 μm ± 0.61), Phe distribution volume, Phe pool size and rate constants between dogs when the tracer was administered IV or orally (p > 0.25). Decay curve for plasma l ‐[1‐¹³C]Phe differed between IV and oral dosing protocols with IV dosing fit best using a two‐compartment model. Phe disappeared from plasma at a mean rate of 2.8%/min. Estimates of gastric emptying and splanchnic extraction did not differ based on oral or IV tracer dosing when the decay curves were fit with the two‐compartment model (p > 0.40). The half‐life for gastric emptying was 18 min, and first‐pass Phe extraction by the splanchnic bed was 24% of the dietary Phe. These results suggest that oral isotope dosing can be used as an alternative to IV isotope dosing in studies that utilize a primed, constant dosing approach to measure protein and amino acid kinetics.     

Renal energy excretion of horses depends on renal hippuric acid and nitrogen excretion

The prediction of renal energy excretion is crucial in a metabolizable energy system for horses. Phenolic acids from forage cell walls may affect renal energy losses by increasing hippuric acid excretion. Therefore, the relationships were investigated between renal energy, nitrogen (N) and hippuric acid excretion of four adult ponies (230–384 kg body weight (BW)) consuming diets based on fresh grass, grass silage, grass cobs (heat‐dried, finely chopped, pressed grass), alfalfa hay, straw, extruded straw and soybean meal. Feed intake was measured; urine and faeces were quantitatively collected for three days. Feed was analysed for crude nutrients, gross energy, amino acids and neutral‐detergent‐insoluble crude protein (CP); faeces were analysed for crude nutrients and cross energy; urine was analysed for N, hippuric acid, creatinine and gross energy. Renal energy excretion (y; kJ/kg BW^0.75) correlated with renal N excretion (x₁; g/kg BW^0.75) and renal hippuric acid excretion (x₂; g/kg BW^0.75): y = 14.4 + 30.2x₁+20.7x₂ (r = .95; n = 30; p < .05). Renal hippuric acid excretion was highest after intake of fresh grass and lowest after intake of soybean meal. The ratio of hippuric acid to creatinine in urine and the excretion of hippuric acid per gram of dry matter intake was significantly higher for fresh grass than for all other rations. There was no relationship between aromatic amino acid intake and renal hippuric acid excretion. The results of the present study and literature data suggest that feed can be categorized into four groups with regard to the energy losses per gram CP intake: (i) protein supplements (e.g., soybean meal): 4.2–4.9 kJ/g CP intake (ii) alfalfa hay, grains, dried sugar beet pulp: 6.4 kJ/g CP intake, (iii) hay, preserved grass products, straw: 5.2–12.3 kJ/g CP intake (mean 8) and (iv) fresh grass. For group (iii) a negative relationship was observed between renal energy losses per gram of CP and the content of CP or neutral‐detergent‐insoluble CP in dry matter.     

Metabolomic profiling in umbilical venous plasma reveals effects of dietary rumen‐protected arginine or N‐carbamylglutamate supplementation in nutrient‐restricted Hu sheep during pregnancy

Maternal nutrient restriction during pregnancy is a major problem worldwide for human and animal production. Arginine (Arg) is critical to health, growth and reproduction. N‐carbamylglutamate (NCG), a key enzyme in arginine synthesis, is not extensively degraded in rumen. The aim of this study was to investigate ameliorating effects of rumen‐protected arginine (RP‐Arg) and NCG supplementation on dietary in undernourished Hu sheep during gestation. From day 35 to 110 of gestation, 32 Hu ewes carrying twin foetuses were randomly divided into four groups: a control (CG) group (n = 8; fed 100% National Research Council (NRC) requirements for pregnant sheep), a nutrient‐restricted (RG) group (n = 8; fed 50% NRC requirements, which included 50% mineral–vitamin mixture) and two treatment (Arg and NCG) groups (n = 8; fed 50% NRC requirements supplemented with 20 g/day RP‐Arg or 5 g/day NCG, which included 50% mineral–vitamin mixture). The umbilical venous plasma samples of foetus were tested by ¹H‐nuclear magnetic resonance. Thirty‐two differential metabolites were identified, indicating altered metabolic pathways of amino acid, carbohydrate and energy, lipids and oxidative stress metabolism among the four groups. Our results demonstrate that the beneficial effect of dietary RP‐Arg and NCG supplementation on mammalian reproduction is associated with complex metabolic networks.     

Pharmacokinetics of harpagoside in horses after intragastric administration of a Devil's claw (Harpagophytum procumbens) extract

Devil's claw is used for the treatment of inflammatory symptoms and degenerative disorders in horses since many years, but without the substantive pharmacokinetic data. The pharmacokinetic parameters of harpagoside, the main active constituent of Harpagophytum procumbens DC ex Meisn., were evaluated in equine plasma after administration of Harpagophytum extract FB 8858 in an open, single‐dose, two‐treatment, two‐period, randomized cross‐over design. Six horses received a single dose of Harpagophytum extract, corresponding to 5 mg/kg BM harpagoside, and after 7 days washout period, 10 mg/kg BM harpagoside via nasogastric tube. Plasma samples at certain time points (before and 0–24 hr after administration) were collected, cleaned up by solid‐phase extraction, and harpagoside concentrations were determined by LC‐MS/MS using apigenin‐7‐glucoside as internal standard. Plasma concentration‐time data and relevant parameters were described by noncompartmental model through PKSolver software. Harpagoside could be detected up to 9 hr after administration. C_max was found at 25.59 and 55.46 ng/ml, t_1/2 at 2.53 and 2.32 hr, respectively, and t_max at 1 hr in both trials. AUC_0–inf was 70.46 and 117.85 ng hr ml^?1, respectively. A proportional relationship between dose, C_max and AUC was observed. Distribution (V_z/F) was 259.04 and 283.83 L/kg and clearance (CL/F) 70.96 and 84.86 L hr^?1 kg^?1, respectively. Treatment of horses with Harpagophytum extract did not cause any clinically detectable side effects.     

A simple estimation of ideal profile of essential amino acids and metabolizable energy for growing Japanese quail

An experiment was conducted to determine apparent metabolizable energy (AME) and amino acid requirements of growing Japanese quail based on ideal protein concept using artificial neural network and desirability function (D‐ANN). Seven‐day‐old quail chicks were assigned to nine experimental diets based on central composite design (CCD) containing five levels of AME (2809–3091 kcal/kg) and CP (19–24.8% of diet). The ratio of lysine (Lys) to CP was set at 0.053 among all treatments, and remaining essential amino acids (EAA) were adjusted to Lys. The experimental data of CCD were fitted to D‐ANN model to compute the optimal values for independent variables. The optimal values of inputs including AME, CP, digestible Lys (dLys), methionine (dMet), total sulphur amino acids (dTSAA), threonine (dThr), tryptophan (dTrp), isoleucine (dIle), valine (dVal) and arginine (dArg) for maximizing gain and minimizing feed conversion ratio were estimated at 2865 kcal/kg, 25, 1.32, 0.55, 0.88, 0.84, 0.20, 0.75, 1.04 and 1.45% of diet, respectively, with D (desirability function) = 0.94. The corresponding optimal amounts of amino acids based on total amino acids were 1.42, 0.59, 0.95, 0.90, 0.22, 0.81, 1.12 and 1.56% of diet respectively. The ideal pattern of essential amino acids to Lys was as follows: dMet: dLys = 0.42, dTSAA: dLys = 0.67, dThr: dLys = 0.64, dTrp: dLys = 0.15, dIle: dLys = 0.57, dVal: dLys = 0.79 and dArg: dLys = 1.09. The results of this study showed that amino acid requirements of modern quails might be higher than those reported by NRC.     

Determination of net energy content of soybean oil fed to growing pigs using indirect calorimetry

The objectives of this experiment were: (i) to determine the net energy (NE) of soybean oil (SBO) fed to growing pigs using indirect calorimetry (IC); and (ii) to evaluate the effects of inclusion rate of SBO on heat production, oxidative status and nutrient digestibility in growing pigs. Eighteen growing barrows were allotted to three diets based on completely randomized design with six replicate pigs (period) per diet. Diets included a corn‐soybean meal basal diet and two test diets containing 5% or 10% SBO at the expense of corn and soybean meal. During each period, pigs were individually housed in metabolism crates for 14 days, including 7 days to adapt to feed, metabolism crate and environmental conditions. On day 8, pigs were transferred to the open‐circuit respiration chambers for measurement of daily O₂ consumption and CO₂ and CH₄ production. During this time, pigs were fed one of the three diets at 2.4 MJ metabolizable energy/kg body weight (BW)^0.6/day. Total feces and urine were collected and daily total heat production (THP) was measured from days 9 to 13 and fasted on day 14 to evaluate their fasting heat production (FHP). The results show that trends of decreased apparent total tract digestibility of neutral detergent fiber (linear, P = 0.09) and acid detergent fiber (linear, P = 0.07) were observed as the content of dietary lipids increased. The average THP for the three diets were 1326, 1208 and 1193 kJ/kg BW^0.6/day, respectively. The FHP of pigs averaged 843 kJ/kg BW^0.6/day and was not affected by diet characteristics. A reduction of the respiratory quotients in the fed state as the inclusion level of SBO increased was observed. In conclusion, the NE values of SBO we determined by indirect calorimetry were 33.45 and 34.05 MJ/kg dry matter under two inclusion levels. THP could be largely reduced when SBO is added in the feed, but the THP of SBO included at 5% in a corn‐soybean meal diet is not different from the THP of SBO included at 10%.     

The effects of dose and diet on the pharmacokinetics of omeprazole in the horse

This study aimed to investigate the effect of diet and dose on the pharmacokinetics of omeprazole in the horse. Six horses received two doses (1 and 4 mg/kg) of omeprazole orally once daily for 5 days. Each dose was evaluated during feeding either a high‐grain/low‐fibre (HG/LF) diet or an ad libitum hay (HAY) diet in a four‐way crossover design. Plasma samples were collected for pharmacokinetic analysis on days 1 and 5. Plasma omeprazole concentrations were determined by ultra‐high pressure liquid chromatography–mass spectrometry. In horses being fed the HG/LF diet, on day 1, the area under the curve (AUC) and maximal plasma concentration (C_max) were higher on the 4 mg/kg dose than on the 1 mg/kg dose. The AUC was higher on day 5 compared to day 1 with the 4 mg/kg dose on the HG/LF diet. On days 1 and 5, the AUC and C_max were higher in horses being fed the HG/LF diet and receiving the 4 mg/kg dose than in horses being fed the HAY diet and receiving the 1 mg/kg dose. These findings suggest that both dose and diet may affect pharmacokinetic variables of omeprazole in the horse.     

Nitrogen Retention and Plasma Amino Acid Responses in Mature Geldings Fed Three Dietary Concentrations of Lysine

Six mature Quarter Horse geldings (age, 11.5 ± 4.7 years; body weight [BW], 526 ± 9.2 kg) were used in a replicated 3 × 3 Latin square design to determine the effects of three dietary lysine (Lys) levels on nitrogen (N) retention and plasma amino acid (AA) concentrations. The geldings were fed a basal diet of concentrate and Bermuda grass hay supplemented with synthetic essential amino acids (EAA) to meet estimated requirements for EAA. Geldings were fed one of three dietary treatments: (1) a basal diet deficient in Lys (L−; 0.027 g of Lys•kg⁻¹ BW•d⁻¹); (2) a basal diet supplemented with synthetic Lys to meet National Research Council (NRC; 2007) requirements (L+; 0.036 g of Lys•kg⁻¹ BW•d⁻¹); or (3) a basal diet supplemented at twice the recommended Lys requirement (2 × L; 0.070 g Lys•kg⁻¹ BW•d⁻¹). Horses fed the 2 × L diet had higher N intakes (P = .0056) than horses fed either the L− or L+ diet. However N retention (P = .63) was not different between treatments. Plasma Lys was greater (P < .0001) in 2 × L than L− and L+ diets. Plasma threonine (Thr; P < .01), methionine (Met; P = .03), and total plasma non-EAA (P < .05) concentrations decreased as dietary Lys increased. These results suggest N retention is not a good response criterion for evaluating the AA requirements of mature horses. However, plasma AA data indicated more efficient use of Thr and Met when horses were fed dietary Lys in concentrations greater than the current requirement recommended by the Nutrient Requirements of Horses (National Research Council, 2007).     

Physicochemical Interpretation of Acid‐Base Abnormalities in 54 Adult Horses with Acute Severe Colitis and Diarrhea

Background

The quantitative effect of strong electrolytes, pCO₂, and plasma protein concentration in determining plasma pH and bicarbonate concentrations can be demonstrated with the physicochemical approach. Plasma anion gap (AG) and strong ion gap (SIG) are used to assess the presence or absence of unmeasured anions.

Hypotheses

The physicochemical approach is useful for detection and explanation of acid‐base disorders in horses with colitis. AG and SIG accurately predict hyperlactatemia in horses with colitis.

Animals

Fifty‐four horses with acute colitis and diarrhea.

Methods

Retrospective study . Physicochemical variables were calculated for each patient. ROC curves were generated to analyze sensitivity and specificity of AG and SIG for predicting hyperlactatemia.

Results

Physicochemical interpretation of acid‐base events indicated that strong ion metabolic acidosis was present in 39 (72%) horses. Mixed strong ion acidosis and decreased weak acid (hypoproteinemia) alkalosis was concomitantly present in 17 (30%) patients. The sensitivity and specificity of AG and SIG to predict hyperlactatemia (L‐lactate > 5 mEq/L) were 100% (95% CI, 66.4–100; P < .0001) and 84.4% (95% CI, 70.5–93.5 P < .0001). Area under the ROC curve for AG and SIG for predicting hyperlactatemia was 0.95 (95% CI, 0.86–0.99) and 0.93 (95% CI, 0.83–0.99), respectively.

Conclusion and Clinical relevance

These results emphasize the importance of strong ions and proteins in the maintenance of the acid‐base equilibria. AG and SIG were considered good predictors of clinically relevant hyperlactatemia.     

Effects of deficiency and surplus dietary threonine on reproductive performance of primiparous pregnant gilts

Dietary threonine imbalance is known to impair reproductive performances of gestating sows, but the underlying mechanisms are largely unknown. In this study, effects of deficiency and surplus dietary threonine during gestation on reproductive performance, serum metabolites and hormones concentration, and colostral nutrient and immunoglobulin contents of primiparous sows were investigated. Ninety primiparous pregnant gilts were assigned to one of the three dietary treatments with different standardized ileal digestible threonine/lysine ratios at 0.59, 0.72 and 0.85, which represented deficient (DT), adequate (AT) and surplus (ST) dietary threonine concentration respectively. Maternal body weight gain from day 80–110 of gestation was highest (p < .05) for gilts fed AT than for gilts fed DT or ST. On days 30 and 110, serum threonine concentration increases in a dose‐dependent manner with the increasing of dietary threonine concentration in (p < .01), serum urea nitrogen concentration was lower (p < .01) in gilts fed AT than DT or ST, and serum insulin‐like growth factor‐I (IGF‐I) was lowest (p < .05) for gilts fed DT. On day 110, gilts fed AT had lower serum progesterone concentration but higher concentrations of serum prolactin (p < .05) compared to DT and ST. Concentration of colostral immunoglobulin A and G from gilts fed DT was lower (p < .05) compared with gilts fed AT or ST. In conclusion, gilts with the adequate threonine intake were more able to conserve dietary amino acids to support foetal and maternal tissue gain. Deficient or ST threonine intake may induce a delay in changes in progesterone and prolactin concentrations during the prepartum period impeding the transition from pregnancy to lactation.     

Threonine incorporation in tissues of growing broiler chickens using L-[15N] threonine

Most amino acid requirement trials appear for whole-body responses, but there is little information concerning amino acid incorporation in individual tissues, which may vary according to the age. L-[¹⁵N] threonine was used to evaluate its incorporation rate and distribution among broiler tissues in different ages. Seventy-two male broiler chickens were distributed into three different phases: starter (4 to 9 days old), grower (18 to 23 days old) and finisher phase (32 to 37 days old). L-[¹⁵N] threonine was added on balanced diets, and birds were fed for five days in each phase. Enriched samples of breast muscle, feathers, liver, jejunum and plasma were collected at 0, 6, 12, 24, 48, 72, 96 and 120 hr after fed birds with the tracer in each phase. In the tissues were analysed dry matter, nitrogen and stable nitrogen. The ¹⁵N isotope abundance according to the time was fitted into exponential or linear equations using a same intercept. The ratio of the steepness or slope coefficients was determined to compare the L-[¹⁵N] threonine incorporation according to the age. In addition, L-[¹⁵N] threonine mass balances were performed to assess the L-[¹⁵N] threonine distribution among the evaluated tissues. Except for feathers, the L-[¹⁵N] threonine incorporation rate decreased with ageing. Taking into account the L-[¹⁵N] threonine distribution in the tissues, only in the jejunum was not observed an increase as the broiler grew. The L-[¹⁵N] incorporation varied in each tissue and according to the age of the broiler chickens. These outcomes could be useful to comprehend changes in amino acid requirements tissue-specific according to age.     

Dietary mannoheptulose does not alter glucose or lipid metabolism in adult Labrador Retrievers

Mannoheptulose (MH), a glycolytic inhibitor, has been preliminarily investigated as a novel functional food ingredient for dogs. This study aimed to determine the effects of dietary MH, delivered as an extract of un‐ripened avocados, on fatty acid and glucose kinetics in healthy adult Labrador Retriever dogs (n = 12 dogs). The study was a double‐blindcrossover with each dog receiving both dietary treatments, control (CON) and MH (400 mg/kg of diet), in random order. Glucose and glycerol plasma turnover (Ra) and oxidation (Ox) were measured in fasting and in response to repeated meal feeding (“fed”) with stable isotope tracers (U‐¹³C‐glucose, 1,1,2,3,3‐D₅‐glycerol) and indirect calorimetry. Palmitate Ra and Ox were examined during repeated meal feeding only using an oral bolus of U‐¹³C‐K₂‐palmitate and indirect calorimetry. MH had no discernible effect on fasting glucose Ra (677, 722 SEM 36 μmol/min, CON, MH) or Ox (107, 109 μmol/min, CON, MH SEM 10 μmol/min) or fed glucose Ra (2913, 3626 SEM 644 μmol/min, CON, MH) or Ox (951, 936 SEM 174 μmol/min, CON, MH). Glycerol Ra, an index of the rate of lipolysis, was not different between dietary treatments (Fast 162, 113 SEM 35 μmol/min CON, MH; Fed 172, 135 SEM 21 μmol/min, CON, MH). Similarly, palmitate oxidation was not impacted by MH feeding (1966, 2276 SEM 79 μmol/min, CON, MH). Together, these findings do not support MH as a novel functional food ingredient at least at the dietary dose tested.