Interactions of energy and predicted metabolizable protein in determining nitrogen efficiency in the lactating dairy cow

Lactating cows are relatively inefficient in converting dietary N to milk N compared with the efficiency of N use for growth in simple-stomached animals. The majority of productive N losses occur in the postabsorptive system. The aim of the study was to test whether predicted metabolizable protein (MP) and dietary energy exerted independent effects on milk protein synthesis and postabsorptive N efficiency. If true, postabsorptive N efficiency would be expected to be greater when animals are fed high-energy diets. Forty mid-lactation cows (32 multiparous Holstein and 8 primiparous Holstein × Jersey crossbreds) were used in a complete randomized design with a 2 × 2 factorial arrangement of diets. Cows were assigned to 1 of 4 dietary treatments: high-energy, high-protein (HE/HP); high-energy, low-protein (HE/LP); low-energy, high-protein (LE/HP); and low-energy, low-protein (LE/LP). Energy concentrations were 1.55 (HE/HP and HE/LP) or 1.44 (LE/HP and LE/LP) Mcal of net energy for lactation (NE_L)/kg of dry matter (DM). Changes in predicted MP were achieved by feeding diets with 6.6 (HE/HP and LE/HP) or 4.6% (HE/LP and LE/LP) ruminally undegradable protein (DM basis). Ruminally degradable protein was held constant at 10.1% of DM. All cows were fed the HE/HP diet from d 1 to 21 followed by the respective treatments from d 22 to 43 (n = 10). Milk protein yield was reduced as dietary energy was reduced. Milk yield followed a similar pattern as milk protein yield. There was a trend for decreased milk yield as crude protein was reduced. There were no interactions between dietary energy and protein for either milk or protein yield. Plasma amino acid concentrations were not affected by treatment. Milk urea N was affected by energy and protein with a significant interaction (HE/HP = 17.2, HE/LP = 12.2, LE/HP = 21.0, LE/LP = 12.2 mg/dL). Nitrogen efficiency calculated from predicted MP supply was affected by energy and protein supplies with no apparent interaction and ranged from a low of 31% (LE/HP) to a high of 43% (HE/LP). The National Research Council model would predict N efficiency more accurately if a representation of the effects of energy on N efficiency were included in the postabsorptive system.     

Effects of rumen-protected methionine and other essential amino acid supplementation on milk and milk component yields in lactating Holstein cows

Objectives of this study were to investigate the effects of supplementing rumen-protected methionine (RP-Met), threonine (RP-Thr), isoleucine (RP-Ile), and leucine (RP-Leu) individually or jointly to a low-protein diet, on the performance of lactating dairy cows, as well as to determine the effects of these amino acids (AA) on the mammalian target of rapamycin (mTOR) in vivo. Ten lactating Holstein cows were randomly allocated to a repeated 5 × 5 Latin square experiment with five 19-d periods. Treatments were high-protein diet (16% crude protein, positive control; HP), low-protein diet (12% crude protein, negative control; LP), LP plus RP-Met (LPM), LP plus RP-Met and RP-Thr (LPMT), and LP plus RP-Met, RP-Thr, RP-Ile, and RP-Leu (LPMTIL). The dry matter intakes (DMI) of the LP, LPM, and LPMT diets were lower than that of the HP diet, whereas the DMI of the LPMTIL diet was intermediate between the HP diet and the other LP diets. Supplementing RP-Met to the LP diet increased the yields of milk and milk protein, increased the content of milk urea N, and tended to increase milk N efficiency. Co-supplementation of RP-Thr with RP-Met resulted in no further milk production increase. Co-supplementation of all 4 rumen-protected amino acids (RP-AA) increased milk and lactose yields to the level of the HP diet and tended to increase milk protein yield compared with the LPMT diet. We found no significant differences in the contents and yields of milk components between the LPMTIL and HP diets except for a lower milk urea N content in the LPMTIL diet. Venous concentrations of the measured AA were similar across the LP and LP diets supplemented with RP-AA. Relative to levels of the HP diet, LP diets had higher venous concentrations of Met and Gly and tended to have higher Phe concentration and lower concentrations of Val and BCAA. The LPMTIL diet had higher venous concentrations of Arg, Lys, Met, Phe, and Glu, and a lower Val concentration. Phosphorylation status of the measured mTOR components in LPM and LPMT treatments were similar to those in the LP treatment but phosphorylation status of mTOR and eIF4E-binding protein 1 (4eBP1) in LPMTIL treatment were higher. The phosphorylation rates of eukaryotic elongation factor 2 (eEF2) in the 4 LP and LP plus RP-AA diets were higher than that of the HP diet. Overall, results of the present study supported the concept that under the relatively short time of this experiment, supplementing RP-AA, which are believed to stimulate the mTOR signal pathway, can lead to increased milk protein yield. This increase appears to be due to increased DMI, greater mTOR signaling, and greater eEF2 activity.     

Effect of protein level and methionine supplementation on dairy cows during the transition period

Cows experience a significant negative protein balance during the first 30 d of lactation. Given the functional effects of AA on health, especially in challenging periods such as calving, higher levels of protein and specific AA in the diet may act to improve health and feed intake. The response of dairy cows to 3 protein supplementation strategies during the transition period and through the first 45 d in milk was evaluated. The final data set had 39 Holstein cows blocked based on parity (primiparous vs. multiparous) and expected calving and randomly assigned within each block to one of 3 dietary treatments: low protein (LP), high protein (HP), or high protein plus rumen-protected methionine (HPM). Treatments were offered from d ?18 ± 5 to 45 d relative to parturition. Pre- and postpartum diets were formulated for high metabolizable protein (MP) supply from soybean meal, and HP and HPM provided higher MP balance than LP. Preplanned contrasts were LP versus HP+HPM and HP versus HPM. Significance was declared at P ≤ 0.05 and trends at 0.05 < P ≤ 0.10. Cows fed HP and HPM had greater fry matter intake (DMI) prepartum than LP (+2 kg/d), and there was a trend for greater DMI with HPM than with HP (+1.6 kg/d). Body weight and condition score before and after calving did not differ among treatments. High protein (HP and HPM) tended to increase milk yield during the first 45 d of lactation (+1.75 kg/d), increased milk lactose content and urea-N in milk and plasma, tended to increase blood BHB 14 d postpartum, and tended to reduce milk/DMI compared with LP. Blood concentrations of calcium at calving and of glucose, and nonesterified fatty acids pre- and postpartum did not differ. High protein induced lower concentration of plasma IL-1 at calving and lowered blood lymphocytes 21 d postpartum, suggestive of a reduced inflammatory status compared with LP. The concentrations of IL-10, tumor necrosis factor alpha, and other hemogram variables did not differ among treatments. Addition of rumen-protected methionine to the HP diet did not alter milk yield but increased fat and total solids concentrations. The rumen-protected methionine had no effect on blood metabolites and immunity markers, with the exception of increased pre-partum insulin concentrations. The data indicate that dairy cows around calving respond positively to an increase in the supply of MP and to rumen-protected methionine supplementation of the HP diet by increasing intake and improving immune status.     

Form of rumen-degradable carbohydrate and nitrogen on microbial protein synthesis and protein efficiency of dairy cows

Sixteen multiparous lactating Holstein cows (four with rumen cannulae) were fed diets varying in the content and form of ruminally degradable carbohydrates and N to examine dietary effects on microbial protein synthesis (MPS) and whole animal N efficiency, and to evaluate the use of a model based on milk urea N (MUN) for predicting urinary N excretion and N utilization efficiency (NUE). A replicated Latin square design (consisting of diet and experimental period) was employed. The four diets consisted of two low protein diets with either 20% ground corn (diet LP) or 13.5% ground corn plus 3% sucrose (diet LP sucrose) and two high protein diets with 13.5% corn and 3% sucrose with either urea (diet HP urea) or soybean meal (diet HP SBM) as supplemental rumen-degradable protein sources. The intakes of dry matter and N were increased by increasing dietary crude protein (CP) level. However, the yields of milk and milk protein were not affected by CP level. Yield of microbial protein was reduced by sucrose and increased by CP level. There were no differences between urea and SBM supplementation on DM intake, milk yield, or MPS. Mean urinary N excretion for all cows (252 g/d) was underestimated by 55 g/d or overestimated by 25 or 33 g/d using alternative equations based on MUN. Subsequently, NUE (mean = 22.4%) was underestimated by 7.5, 3.2, or 2.9%, using a previously published set of equations. Urinary N excretion and NUE could be predicted within 10 and 14% of observed values, respectively, using a set of equations incorporating MUN. Therefore, MUN appears to be a useful tool to help assess N losses from lactating cows.     

Long-term high animal protein diet reduces body weight gain and insulin secretion in diet-induced obese rats

BACKGROUND: The effects of a high protein diet on insulin secretion and glucose metabolism have been quite controversial. The aim of this study was to evaluate the effects of long‐term isocaloric high animal protein intake on insulin secretion in diet‐induced obese rats. RESULTS: After the experimental period (24 weeks), the high‐fat diet‐induced obese rats that were fed isocaloric high‐protein diets (HP) had lower body weight gain (P < 0.01) and lower visceral fat (P < 0.05) than normal protein (NP) rats. Fasting plasma glucagon‐like peptide‐1 (GLP‐1) was also reduced significantly (P < 0.05), as well as serum insulin levels at 5 min and 10 min by intravenous insulin releasing test. In addition, insulin mRNA and pancreatic duodenal homeodomain‐1 (PDX‐1), GLP‐1 protein expression were both markedly lower in HP rats (P < 0.05), while PDX‐1 mRNA in HP rats had no difference from NP rats. CONCLUSION: These results suggest that long‐term isocaloric high animal protein intake reduces the acute insulin response in obese rats and the decrease of insulin is associated with both reduced weight gain and inhibition of PDX‐1 expression. GLP‐1 might be a negative feedback for the balance of energy metabolism secondary to changes of body weight and visceral fat. Copyright © 2012 Society of Chemical Industry     

The effect of protein intake on performance of cows in hot environmental temperatures.

Two trials were conducted with cows in commercial herds during midlactation to evaluate the effect of dietary crude protein (CP) concentration on the production, composition, and efficiency of milk production under hot ambient conditions. Cows were group-fed in trial 1, which was conducted in two herds, and were fed individually in trial 2. The respective average ambient temperature, relative humidity, and temperature-humidity index (THI) were 31 degrees C, 45%, and 78 in trial 1 and 27 degrees C, 70%, and 76 in trial 2. Cows were cooled by forced evaporative means six times daily in trial 1 and three times daily in trial 2. Dietary CP was 15.3 or 17.3% of dry matter (DM) in trial 1 and 15.1 or 16.7% of DM in trial 2. The respective ratios of rumen-degradable organic matter (RDOM) to rumen-degradable protein were 5.3 and 4.8 for the low CP (LP) and high CP (HP) diets. Average DM intake, milk yield, and milk fat and protein concentrations were 22 and 23 kg/d, 34 and 35 kg/d, 3.1 and 3.4%, and 3.2 and 3.1% in trials 1 and 2, respectively, and were similar among diets in both trials. The resultant calculated milk protein efficiency ratio and overall CP efficiency were 0.31 and 0.32 for the LP diets and 0.28 and 0.29 for the HP diets. In cows fed the LP diet, diet rumen ammonia was lower in trial 1, and milk urea N was lower in trial 2. The BW change was higher in trial 1, and tended to be higher in trial 2, with the LP diets. Changes in body condition score in trials 1 and 2 tended to be higher with the LP diets. It was concluded that a dietary CP content of 15.3% is adequate to maintain production in heat-exposed dairy cows producing 35 kg of milk/d, provided that the forced evaporative cooling and the ratio of RDOM to rumen-degradable protein is appropriate     

Effect of feeding complete feed block containing Prosopis cineraria leaves and polyethylene glycol (PEG)‐6000 on nutrient intake,its utilization,rumen fermentation pattern and rumen enzyme profile in kids

A study was carried out to determine the effect of replacing the cake portion of concentrate mixture with 5 parts polyethylene glycol (PEG)‐6000 in a complete feed block (CFB) containing Prosopis cineraria leaves on the performance, rumen fermentation pattern and rumen enzyme profile of kids under an intensive system of rearing. Eighteen weaners of Sirohi goat of similar body weight (16.0 ± 0.5 kg) and age (90 ± 5 days) were divided into three equal groups. They were housed in individual cages in a side open asbestos roof shed with mud floor. All the kids received CFBs containing 50 parts P cineraria leaves and 50 parts concentrate mixture. CFB offered to the first group had high protein (183.8 g kg^?1; HP), that offered to the second group had low protein (131.3 g kg^?1; LP) and that offered to the third group had low protein (124.7 g kg^?1) but contained PEG‐6000 (LP‐PEG). The concentrate mixtures in LP and LP‐PEG were without groundnut cake, whereas in LP‐PEG, groundnut cake was replaced by barley and 5 parts PEG‐6000 were incorporated. CFBs were similar in their nutritive value except for crude protein (CP). Prosopis leaves utilized in the CFB contained (g kg^?1 dry matter) 129.1 CP, 535.5 neutral detergent fibre, 395.8 acid detergent fibre and 222.8 acid detergent lignin. There were significant differences in dry matter intake (g day^?1) between HP (1102), LP (1108) and LP‐PEG (1194); the trend in Metabolizable energy intake was similar. During the growth trial, LP kids consumed maximum amount of feed (76.91 kg) followed by HP (75.73 kg) and LP‐PEG (73.12 kg). However, maximum feed efficiency (feed consumed kg^?1 live weight gain) was recorded in LP‐PEG kids (9.59) followed by HP (10.64) and LP (11.60). These differences were statistically significant (p < 0.05). Although there was no significant difference in the digestibility of dry matter among the groups, there was significant difference in the digestibility of CP, neutral detergent fibre and acid detergent fibre. The digestibility of CP was 0.591, 0.484 and 0.645, that of neutral detergent fibre was 0.397, 0.308 and 0.499 and that of acid detergent fibre was 0.168, 0.154 and 0.282 in HP, LP and LP‐PEG, respectively. Rumen metabolites studied 6 h after feeding revealed that there were significant (p < 0.05) differences in the concentrations of ammonia N, tri‐chloro acetic acid precipitable N and total volatile fatty acids among the three groups, but not pH. The rumen enzyme concentrations showed significant (p < 0.05) differences for α‐amylase, whereas the differences were non‐significant for CMcase and protease. After 90 days of feeding trial, the maximum weight gain was recorded in LP‐PEG (7.62 kg) followed by HP (7.23 kg) and LP (6.53 kg). It can be concluded that, when kids are reared under an intensive system on complete feed blocks containing Prosopis leaves, high protein concentrate can be replaced with a low protein concentrate containing 5 parts PEG, which would not only alleviate the negative effects of tannin but would also spare expensive groundnut protein. Copyright © 2005 Society of Chemical Industry     

Protein nutrition of dairy cows receiving grass silage diets. Effects on silage intake and milk production of postruminal supplements of casein or soya-protein isolate and the effects of intravenous infusions of a mixture of methionine,phenylalanine and tryptophan

Milk production responses to protein supplementation of grass silage diets were investigated in two experiments with dairy cows. In Experiment 1. four cows received a basal diet of grass silage ad libitum plus a barley/soya supplement at 5 kg day^?1. The cows received the following treatments in a 4 × 4 Latin square design with 10-day periods: (1) basal diet, (2) basal diet plus 230 g casein day^?1 infused into the abomasum; (3) basal diet plus 225 g soya protein isolate (SPI) day^?1 into the abomasum; and (4) basal diet plus a dietary addition of 1.2 kg fishmeal-based product (AP) day^?1. All three supplements increased (P < 0.05) the yield of milk and milk protein relative to the basal diet, the values being 16.9. 535:18.8. 610; 19.5, 625; and 18.5 kg day^?1 and 582 g day^?1 for the basal. AP, casein and SPI treatments respectively. Responses to casein were greater (P < 0.05) than to SPI. Changes in the concentrations of amino acids in blood plasma suggested that methionine, tryptophan and phenylalanine were in shortest supply relative to demand, and their potential role as limiting amino acids was investigated in a second experiment. In Experiment 2, four dairy cows were used in a 4 × 4 Latin square design with 10-day periods. The four treatments were (1) a diet of grass silage ad libitum plus 5 kg day^?1 of a barley/soya supplement containing 176 g CP kg^?1 DM (LP); (2) LP plus an intravenous infusion of 5.0 g methionine, 9.1 g phenylalanine and 2.2 g tryptophan day^?1; (3) grass silage ad libitum plus 5 kg day^?1 of a barley/soya supplement containing 334 g CP kg^?1 DM (HP); and (4) HP plus the intravenous infusion of methionine, phenylalanine and tryptophan. The HP treatment produced a higher yield (P < 0.05) of milk protein than the LP treatment (722 vs. 667 g day^?1) but the amino acid infusion had no effect with either treatment. The elimination of methionine, phenylalanine and tryptophan (and, in previous experiments, methionine and lysine) as candidates for limiting amino acids on diets of grass silage plus soya-based supplements questions whether the differences in responses to casein and SPI can be explained in terms of their amino acid compositions. The role of factors such as peplides, producing during digestion, some of which may be biologically active, deserves investigation.     

Influence of calcium and phosphorus feeding on markers of bone metabolism in transition cows

A study was carried out to verify the effect of Ca and P levels on production, digestibility, and serum bone metabolism biomarkers in dairy cows. Fifty-two nonlactating multiparous cows (≥3 lactations) were confined in a free-stall barn approximately 20 d before calving. A standard close-up diet was fed to cows once daily until d 2 postpartum. Cows were randomly assigned to 1 of 4 dietary treatments arranged in a 2 × 2 factorial approach averaging 0.64% Ca for high Ca (HCa), 0.46% Ca for low Ca (LCa), 0.47% P for high P (HP), and 0.38% P for low P (LP) on a dry matter basis. Experimental diets were fed twice daily from 3 d in milk (DIM) until 31 DIM. Intake and milk yield were recorded daily. Milk samples were collected on d 28, 29, and 30 postpartum for components analyses. Blood samples were drawn 10 d before expected calving, at calving, and at 15 and 30 DIM for serum analyses of osteocalcin, a biomarker of bone accretion, and pyridinoline, a biomarker of bone resorption. Total fecal collection was conducted when cows in a block averaged 20 DIM. Intake and production traits were not significantly affected by any of the dietary treatments. Cows averaged nearly 21 kg/d dry matter intake and 44 kg/d milk yield from 6 to 31 DIM. There were no significant differences across treatments in body weight or body condition score loss. Phosphorus intake, P fecal output, P digestibility, and P apparent absorption were affected by dietary P content. Calcium intake was higher with HCa, but Ca fecal output, digestibility, and apparent absorption showed an interaction between dietary Ca and dietary P. Calcium fecal output was 100.6 g/d for cows fed HCaHP, intermediate for cows on the HCaLP diet (89 g/d), and similar among cows fed the 2 LCa diets (70 g/d with LCaHP and 75 with LCaLP). There was no significant effect of Ca or P on osteocalcin measurements. Pyridinoline concentrations were affected by dietary Ca levels and tended to have a significant dietary Ca × dietary P interaction. Phosphorus apparent digestibility occurred independently of dietary Ca levels. Results of this study suggest that more bone was mobilized in cows fed LCa diets, but excess dietary P caused greater and prolonged bone mobilization regardless of dietary Ca content.     

Amino acid efficiencies of utilization vary by different mechanisms in response to energy and protein supplies in dairy cows: Study at mammary-gland and whole-body levels

Variations of mammary gland (MG) metabolism were studied in dairy cows in response to diets containing 2 levels of net energy of lactation [NE_L; 25.0 and 32.5 Mcal/d for low (LE) and high energy (HE), respectively], combined with 2 levels of metabolizable protein [MP, 1,266 and 2,254 g/d of protein digestible in the intestine for low (LP) and high protein (HP), respectively] in a 2 × 2 factorial arrangement. Four cows received 4 diets (LELP, HELP, LEHP, and HEHP) in a 4 × 4 Latin square design with 2-wk experimental periods. Milk production and feed intake were measured on the last 5 d of each period, whereas MG net uptake of AA was determined on d 13. Efficiencies were estimated as the sum of measured milk true protein yield (MPY) and of estimations of metabolic fecal and scurf proteins multiplied by their respective AA profile and divided by the estimated AA supply minus the AA endogenous urinary loss. The increased MPY in the HE compared with the LE diets (higher by 123 g/d) was accompanied by increased mammary plasma flow and MG uptake of the nonessential AA (NEAA) and the essential AA (EAA), except for branched-chain AA. In contrast, the increase in MPY (higher by 104 g/d) observed in the HP compared with the LP diets was linked to increased MG uptake of EAA without a change in mammary plasma flow and a decreased NEAA uptake. Because MG uptake of total AA-N was almost equal to cows' milk output on a nitrogen basis, these different mechanisms involve a large MG flexibility, with variable synthesis of NEAA. In addition, MP efficiency did not increase only through increased MPY in the HE compared with the LE diets but also through metabolic fecal protein, estimated to increase (by 65 g/d) with dry matter intake. The MPY increased in the HP compared with the LP diets, but the increase was smaller than the calculated increase (greater by 993 g/d) in MP supply. The highest MG clearance rates of individual EAA could suggest that Met, His, and Lys were limiting in LP, and Met was the most limiting AA in HP. Interestingly, a similar hypothesis could be stated by analyzing estimated AA efficiencies. The highest efficiencies among EAA, observed for His in HELP and for Met with the other diets, could indicate that they were the most limiting AA in these respective diets, whereas other EAA (including Lys) efficiencies varied with MP efficiency. The MG metabolic flexibility with regard to individual AA utilization partially contributes to the anabolic fate of AA through MPY; however, other export proteins also contribute to variations in MP and AA efficiencies.     

PPARγ Pro12Ala interacts with fat intake for obesity and weight loss in a behavioural treatment based on the Mediterranean diet

Scope: The goal of this study was to examine whether the Pro12Ala polymorphism of peroxisome proliferator‐activated receptor γ (PPARγ) is associated with insulin resistance, obesity and weight loss and to analyze potential interactions between fat intake and PPARγ polymorphism in a Spanish overweight/obese population. Materials and methods: We recruited 1465 subjects enrolled in a behavioural treatment program for obesity based on a Mediterranean diet, which included the following: dietary treatment, physical activity, nutritional education and behavioral techniques. A significant association was found between PPARγ2 Pro12Ala genotype and plasma insulin concentration and homeostasis model assessment insulin resistance. Subjects with the Ala12 genotype had lower insulin levels than those with the Pro12Pro genotype. We detected a gene–diet interaction between the PPARγ Pro12Ala polymorphism and MUFA for BMI and body fat. Furthermore, we detected an interaction between the PPARγ Pro12Ala polymorphism and fat intake for total weight loss (p<0.001). When total fat intake was high, Ala12‐carriers exhibited a significantly lower percentage of total weight loss than major‐allele‐carriers (p=0.037). Conclusion: Data are consistent with previous results showing a protective role for the Ala12 allele against insulin resistance, and replicate an earlier study that detected an interaction between dietary MUFA and PPARγ2 for BMI. Our detection of a gene–diet interaction between PPARγ Pro12Ala and fat intake for weight loss may explain previous discrepancies among different studies.     

Effects of plane of nutrition and feed deprivation on insulin responses in dairy cattle during late gestation

Nonlactating Holstein cows (n=12) in late pregnancy were used to determine effects of plane of nutrition followed by feed deprivation on metabolic responses to insulin. Beginning 48 d before expected parturition, cows were fed to either a high plane (HP) or a low plane (LP) of nutrition (162 and 90% of calculated energy requirements, respectively). Cows were subjected to an intravenous glucose tolerance test [GTT; 0.25 g of dextrose/kg of body weight (BW)] on d 14 of treatment and a hyperinsulinemic-euglycemic clamp (HEC; 1 μg/kg of BW/h) on d 15. Following 24 h of feed removal, cows were subjected to a second GTT on d 17 and a second HEC on d 18 after 48 h of feed removal. During the feeding period, plasma nonesterified fatty acid (NEFA) concentrations were higher for cows fed the LP diet compared with those fed the HP diet (163.6 vs. 73.1 μEq/L), whereas plasma insulin was higher for cows fed the HP diet during the feeding period (11.1 vs. 5.2 μIU/mL). Glucose areas under the curve during both GTT were higher for cows fed the LP diet than for those fed the HP diet (4,213 vs. 3,750 mg/dL × 60 min) and was higher during the GTT in the feed-deprived state (4,878 vs. 3,085 mg/dL × 60 min) than in the GTT during the fed state, suggesting slower clearance of glucose during negative energy balance either pre-or post-feed deprivation. This corresponded with a higher dextrose infusion rate during the fed-state HEC than during the feed-deprived-state HEC (203.3 vs. 90.1 mL/h). Plasma NEFA decreased at a faster rate following GTT during feed deprivation compared with that during the fed state (8.7 vs. 2.9%/min). Suppression of NEFA was highest for cows fed the HP diet during the GTT conducted during feed deprivation, and lowest for cows fed the HP diet during the fed-state GTT (68.6 vs. 50.3% decrease from basal). Plasma insulin responses to GTT were affected by feed deprivation such that cows had a much lower insulin response to GTT by 24 h after feed removal (995 vs. 3,957 μIU/mL × 60 min). During the fed-state HEC, circulating concentrations of NEFA were 21% below basal for cows fed the HP diet and 62% below basal for cows fed the LP diet; during feed deprivation, NEFA were 79 and 59% below basal for the HP and LP diets, respectively (diet × HEC). Cows that are fed below energy requirements or are feed deprived have slower clearance of glucose and greater NEFA responses to glucose challenge. Additionally, feed deprivation had a large effect on insulin secretion. Overall, effects of feed deprivation were larger than effects of plane of nutrition.     

Active Bowman–Birk inhibitors survive gastrointestinal digestion at the terminal ileum of pigs fed chickpea‐based diets

BACKGROUND: Protease inhibitors of the Bowman–Birk family have been demonstrated to be naturally occurring chemopreventive agents in a wide range of in vitro and in vivo models. In vitro and in vivo experiments have reported that Bowman–Birk inhibitors (BBIs) may exert colorectal chemopreventive effects. To exert such effects, these proteins have to survive, at least to some extent, the digestive process within the gastrointestinal tract. RESULTS: In order to determine the survival rates of functional BBI proteins in vivo, five castrated male pigs (100 ± 2 kg body weight) fitted with T‐shaped cannulas at the terminal ileum were fed a chickpea‐based diet. Pigs fed hydrolysed casein as the only protein source were used as an experimental control diet. The survival rates of BBI proteins from chickpea‐based diets at the terminal ileum, expressed in terms of trypsin (TIA) and chymotrypsin inhibitory activity (CIA), were 7.3 and 4.4%, respectively. The presence of BBI proteins in ileal samples from pigs fed chickpea‐based diet was confirmed by SDS‐PAGE and peptide mass fingerprinting. CONCLUSION: It is concluded that significant amounts of active BBI proteins reach the large intestine of the pig. Further pharmacological studies are necessary in order to determine the potential of dietary BBI proteins as colorectal chemopreventive agents in humans. Copyright © 2007 Society of Chemical Industry     

Effects of high dietary crude protein on the characteristics of preovulatory follicles in dairy heifers

The objectives were to examine the effect of high dietary crude protein on characteristics of preovulatory follicles in dairy heifers. Eight Israeli-Holstein heifers, 4 fitted with rumen fistula and 4 intact, were assigned to 1 of 3 treatments in a replicated (n = 2) 4 × 3 incomplete Latin square design with 39-d periods. Treatments were: low (6.0%; LP), moderate (13.0%; MP), and high (20.0%; HP) crude-protein diets, containing 1.27 Mcal NE_L/kg dry matter. Diets were based on approximately 66% wheat straw and various proportions of ground corn grain and soybean meal. The estrous cycles of the heifers were synchronized, and 14 d after behavioral estrus, heifers received PGF_2α injections. After a further 40 h, at d 39 of each period, follicular fluid (FF) was aspirated from follicles of diameter >7 mm. The intake of the LP diet was 9% lower than that of MP and HP diets. Rumen ammonia and plasma urea nitrogen concentrations were highest in the HP and lowest in the LP, with intermediate levels in MP diets. No differences were found between treatments in plasma and FF concentrations of glucose and nonesterified fatty acids. High-protein diets increased urea concentrations very similarly in preovulatory FF and in plasma. No differences were observed between diets, in preovulatory follicle diameters and concentrations of androstenedione. However, higher estradiol and progesterone concentrations in FF were observed under the HP than under the MP diet, with no difference between diets in estrogen to progesterone ratio. It can be concluded that high concentrations of urea in plasma, caused by high dietary crude protein, penetrated into preovulatory follicles, but did not impair preovulatory characteristics. This lack of detrimental effects might be attributed to the use in this study of nonlactating heifers, which have fewer nutritional and physiological constraints and eliminate negative effects of potential interactions with high urea on dairy cows’ reproductive systems.     

Protein level for alfalfa and corn silage-based diets: II. Nitrogen balance and manure characteristics

This N balance study was completed with 48 multiparous Holstein cows (body weight [BW] = 653 kg; days in milk = 89) blocked by calving date and assigned to a 2 x 2 factorial arrangement of dietary treatments. The total mixed ration included alfalfa silage (AS) or corn silage (CS) as the primary forage source (41 and 14% vs. 14 and 41% of diet dry matter (DM), respectively) and were formulated for recommended (RP) or excessive (HP) amounts of rumen degradable protein (RDP) and rumen undegradable protein (RUP) according to the guidelines of the National Research Council (NRC). Crude protein (CP) averaged 16.5, 18.0, 16.4, and 17.3% for the AS-RP; AS-HP; CS-RP; and CS-HP diet, respectively (DM basis). Regardless of primary forage source, the reduction in dietary CP to the NRC guidelines tended to improve milk yield (43.4 vs. 41.0 kg/d) but did not alter 3.5% fat-corrected milk (37.0 kg/d) or milk true protein yield (1167 g/d). In this trial, cows fed the CS-based diets consumed less DM than those fed the AS-based diets in part because of rumen acidosis. The adverse effect of low rumen pH was accompanied by an increase in urinary N (UN) as a percentage of N intake, but did not alter milk yield. Notwithstanding partial confounding, fecal N (FN) was 49 g/d lower (213 vs. 164 g/d), UN was unchanged (229 g/d), but milk N tended to be higher (194 vs. 206 g/d) when cows were fed the CS-based diets compared with AS-based diets. Compared with the HP diets, cows fed the RP diets had similar FN (189 g/d) and milk N (200 g/d), but UN and urine urea N were reduced by 41 g/d (249 vs. 208 g/d) and 40 g/d (210 vs. 171 g/d), respectively. Fecal N concentration was higher for CS-based diets, but urinary N concentration was higher for AS-based diets. The reduction in dietary CP did not influence these concentrations but lowered urine volume. The metabolic relationships between energy and protein in determining the fate of excess dietary N (primarily in the form of excess RUP in this trial) was illustrated by a 17% increase in the UN to FN ratio for cows fed AS-HP compared with the AS-RP diet and a 42% increase in the UN to FN ratio for CS-HP compared with CS-RP diet, when cows' energy status was compromised because of rumen acidosis. In this trial, UN ranged from 150 to 320 g/d, and was best predicted as UN (g/d) = 0.0283 x BW (kg) x milk urea N (mg/dL). The NRC protein guidelines should not be exceeded to avoid unnecessary losses of manure N and, in particular, urine urea N.     

Starch–protein interaction in the rumen of weaned dairy calves

The objectives of this study were to investigate the effect of starch and protein interaction on rumen environment, in situ digestion, and total-tract digestibility of nutrients in weaned dairy calves between 8 and 16 wk of age. Sixteen rumen-cannulated calves were randomly divided into 4 dietary treatment groups with 4 calves fed in each treatment. The treatment diets had 2 levels of starch [18%, low starch (LS), or 38%, high starch (HS)] and 2 levels of protein [16%, low protein (LP), or 22%, high protein (HP)] on a dry matter (DM) basis in calf grower: (1) LPLS, (2) LPHS, (3) HPLS, and (4) HPHS. Calves were fed for ad libitum intake (95% assigned grower and 5% grass hay), and refusals were collected weekly. Total-tract digestibility collection and in situ digestibility procedures were performed for each calf at 11 and 15 wk. Samples for in situ digestibility, grass hay (GH), soybean hulls (SBH), wheat middlings (WM), ground corn (GrC), and soybean meal (SBM) were incubated for 9 and 24 h. There was no starch and protein interaction on total-tract digestibility of calves. Total-tract DM, neutral detergent fiber (NDF) and acid detergent fiber (ADF) digestibility, and feed efficiency were affected by both protein and starch inclusion level in calf diet. Total-tract starch digestibility was lower for LS diets. Dry matter digestibility and feed efficiency were greater in calves fed HP and HS diets compared with calves fed LP and LS diets, respectively. Fiber digestibility (NDF and ADF) was less in calves fed HS diets compared with calves fed LS diets but was greater in calves fed HP diets compared with calves fed LP diets. Level of protein did not affect in situ DM and NDF disappearance of GH, but HP increased in situ DM and NDF disappearance of SBH. High-starch diets decreased DM and NDF disappearance of both GH and SBH. At 20 h after feeding, ruminal pH was 0.51 unit higher in calves fed HPHS compared with calves fed LPHS. Total ruminal VFA and proportion of propionate was greater with HS versus LS, whereas proportion of acetate was greater with LS versus HS. The DM disappearance of SBM and WM and NDF disappearance of WM was greater for calves fed HPHS compared with calves fed LPHS at 11 wk of age. In our study, when HP was fed with HS, rumen pH, in situ digestion of WM and SBM, and total-tract digestion of DM, NDF, and ADF increased. This provides evidence for starch–protein interaction in the rumen of recently weaned dairy calves. Improvements in total-tract and in situ digestibility suggest that both protein and starch levels are important for 8- to 16-wk-old calves.     

EFFECTS OF FEED DIETS ON DIGESTIVE PROTEASES FROM THE HEPATOPANCREAS OF WHITE SHRIMP (Penaeus vannamei)

Protease activities in the hepatopancreas extract (HP) from white shrimp (Penaeus vannamei) fed one of seven test diets for 30 days were evaluated by several methods. The test diet contained 85% of a reference ration for shrimp and 15% of either anchovy meal, tuna waste meal, deboned white fish meal, langostilla meal, soybean meal, and two menhaden meals as a protein replacer. One of the menhaden fish meals tested (B) had the lowest quality as a shrimp feed based on amino acid analysis. SDS-PAGE zymograms of HP from each of the seven diet groups showed similar proteins activity patterns with casein as substrate. The degree of hydrolysis of casein, measured by pH-stat, was also the same for HP from the seven diet groups (P > 0.05). However, total protease activity measured by azocasein hydrolysis (units/g HP) was higher for the diet group fed the test ration containing tuna waste as a replacer (P > 0.05). Trypsin and chymotrypsin activities measured with synthetic substrates (units/mg protein; units/g HP) from animals reared on the diet with menhaden meal B replacer were greater than in the other diet groups (P < 0.05). This study shows that a relatively small amount (15%) of a specific protein replacer in white shrimp rations can influence the protease activity of shrimp HP. Given that digestive proteases such as trypsin can leach into the muscle of postharvest shrimp and thereby cause softening of the meat, the impact of the rearing diet on postharvest shelf-life should be considered along with the standard measures of feed quality that are used by the fish farmer, i.e. animal growth and heatlh.     

Feasibility of fishmeal replacement by shrimp head silage protein hydrolysate in Nile tilapia (Oreochromis niloticus L) diets

This study provides information on the use of shrimp head silage protein hydrolysate (SPH) as an alternative protein source for tilapia feeding. Six diets (28% protein, 12% lipid) were prepared where fishmeal protein was replaced at levels of 0 (control), 10, 15, 20, 25 and 30% with the hydrolysate. The diets were supplied to Nile tilapia fry (338 mg initial weight) stocked in plastic recirculating 20 l tanks (10 animals per tank), with three replicates per treatment. After an 8 week experimental period, fish fed the diets containing 10 and 15% SPH showed significantly better performance in terms of final body weight, weight gain (%), mean daily weight gain (mg day^?1), specific growth ratio and feed conversion ratio than those fed the control diet (fishmeal as protein source) and higher‐SPH diets. It is concluded that shrimp head hydrolysate is a promising alternative protein source for tilapia feeding, improving growth ratio at dietary inclusion levels as high as 15%. In addition, the diets with added shrimp silage protein were well accepted by the fish, which avidly consumed the feed during the experiment. © 2002 Society of Chemical Industry     

The influence of different single dietary sources on moult induction in laying hens

An investigation was carried out to assess the possibility of using single dietary sources as alternatives to feed deprivation for the induction of moult in commercial laying hens. The study involved six dietary groups of 29 laying hens: unmoulted, dried tomato pomace, alfalfa meal, rice bran, cumin seed meal and feed withdrawal. The birds received the above diets during the moulting period (11 days), and body weight loss and ovary weight regression were measured. Post‐moult production parameters (number of eggs produced per hen per day, egg weight, shell weight, yolk colour and Haugh unit) were measured for 12 weeks. Results showed that all dietary sources were as effective as feed withdrawal in causing ovary weight regression in birds. Birds provided with tomato pomace or alfalfa showed lower weight losses than feed‐deprived birds at the end of the moulting period. Hens moulted by tomato pomace or alfalfa exhibited post‐moult levels of egg production over a 12 week period that were superior to those of hens moulted by feed withdrawal. Post‐moult eggs laid by hens moulted by all dietary sources were of comparable quality to eggs from feed‐deprived hens and superior to those from unmoulted hens. As fibrous feeds with low metabolisable energy and an appreciable amount of protein, dried tomato pomace and alfalfa meal may be fed to hens on an ad libitum basis for effective moult induction while reducing the stress of severe starvation and retaining comparable egg quality and production. Copyright © 2007 Society of Chemical Industry     

Effect of supplementary concentrate type on nitrogen partitioning in early lactation dairy cows offered perennial ryegrass-based pasture

Forty-four early lactation (64 ± 20 d in milk) dairy cows of mixed parity were used to assess the effect of 4 supplementary concentrate types (n=11) on N partitioning. Animals were blocked on parity and calving date, and blocks were balanced for previous milk yield and milk protein yield. Cows received grazed pasture plus 5.17 kg of dry matter (DM)/d of one of the following isoenergetic concentrates: high crude protein (CP) with rolled barley (HP, 19% CP); low CP with rolled barley (LP, 15% CP); low CP with barley and supplementary 2-hydroxy-4-methylthio butanoic acid (HMBi; LP+HMBi, 15% CP); and low CP with ground corn (LP Corn, 15% CP). Nitrogen partitioning studies were conducted at wk 6 and 10 postpartum by using the n-alkane technique to determine pasture dry matter intake (DMI). Pasture DMI (13.3kg of DM/d) and dietary digestibility of DM were not affected by concentrate type. Milk yield was lower for LP compared with other concentrate types (25.4 vs. 28.3 kg/d). Yields of milk protein and milk casein were not affected by concentrate type. However, milk solid yield and milk fat yield were higher for LP+HMBi (1.97 and 0.92 kg/d) compared with LP (1.72 and 0.87 kg/d). Concentrations of fat, protein, lactose, and casein were not affected by concentrate type. Dietary N intake was higher for HP compared with other treatments (0.545 vs. 0.482 kg/d, HP vs. average of the 3 LP treatments). Dietary N intakes were not different among low CP concentrates. Fecal N excretion was not affected by concentrate type. However, urinary N excretion was related to N intake and was higher for HP compared with other treatments (0.261 vs. 0.195 kg/d, HP vs. average of the 3 LP treatments). Urinary N excretion was not different among low CP concentrates. Milk N output was higher for HP (0.139 kg/d) compared with LP (0.12 kg/d) but not LP+HMBi (0.137 kg/d) or LP Corn (0.138 kg/d). The portion of feed N excreted as feces N was lower for HP compared with other treatments (0.272 vs. 0.327, HP vs. average of the 3 LP treatmentsHowever, the portion of feed N excreted as urine N was higher for HP (0.466) compared with LP+HMBi (0.408) and LP Corn (0.366) but not compared with LP. The portion of feed N excreted as milk N was higher for LP Corn (0.282) compared with HP (0.257) but not LP+HMBi or LP. Dietary reformulation to reduce N excretion in pasture-based dairy production systems is possible. However, maintenance of milk yield and milk N when concentrate CP was reduced (19 vs. 15%) required the use of either protected AA (HMBi) or ground corn.