Effects of supplementing milk replacer with essential amino acids on blood metabolites, immune response, and nitrogen metabolism of Holstein calves exposed to an endotoxin.

This study evaluated the effects of supplementing calf milk replacer with essential AA on immune responses, blood metabolites, and nitrogen metabolism of 32 Holstein bull calves [28 d of age, 44 ± 0.8 kg of body weight (BW)] exposed to lipopolysaccharide (LPS). Calves were bottle-fed a commercial milk replacer (20% crude protein and 20% fat, dry matter basis) twice daily along with a calf starter (19% crude protein, dry matter basis) for 45 d. The experiment was a randomized complete block design and treatments were a 2 × 2 factorial arrangement. Treatments were milk replacer (fed twice daily at 0.5 kg/d of powder) supplemented with or without 10 essential AA (+AA vs. -AA), and subcutaneous injection of sterile saline with or without LPS (+LPS vs. -LPS) at 3 h after the morning feeding on d 15 (4 µg LPS per kg of BW) and 17 (2 µg LPS per kg of BW). Calves also received a 2-mL subcutaneous injection of ovalbumin (6 mg of ovalbumin/mL) on d 16 and 30. Rectal temperature and blood samples were collected on d 15 before LPS injection and at h 4, 8, 12, and 24 thereafter. From d 15 to 19, total fecal and urinary output were collected, and feed refusals were documented. Rectal temperature was greater in +LPS than -LPS calves at h 4, 8, and 12 after LPS injection. Serum cortisol was greater for +LPS than -LPS at h 4 after LPS exposure. At d 28, serum antiovalbumin IgG level was greater in +LPS +AA calves compared with +LPS -AA. Serum glucose was lower for +LPS than -LPS at h 4 and 8. Serum insulin was greater in +LPS than -LPS calves. Plasma concentrations of Thr, Gly, Asn, Ser, and hydroxyproline were lower for +LPS versus -LPS calves. Plasma concentrations of Met, Leu, Phe, His, Ile, Trp, Thr, and Orn were greater in +AA calves than -AA calves. Plasma urea N and N retention were not different among LPS and AA treatments. The lower concentrations of AA in +LPS than -LPS calves indicate higher demand for AA in immuno-compromised calves fed milk replacer. Additionally, higher concentration of ovalbumin-specific IgG level in +LPS calves supplemented with +AA compared with +LPS calves with -AA suggests that supplementing AA to immune-compromised calves might improve immune status.

Influence of Monensin and Diflubenzuron on the Control of Immature Horn Fly and House Fly (Diptera: Muscidae) and Chemical Composition of Cattle Feces.

Twelve Angus × Hereford crossbred heifers were used in a completely randomized design to evaluate the effects of monensin and diflubenzuron feed additives on horn fly (Haematobia irritans) (L.) and house fly (Musca domestica) (L.) immature development. Heifers were randomly assigned to one of four treatment groups; 1) monensin fed at label rate, 2) diflubenzuron fed at label rate, 3) monensin and diflubenzuron fed at label rates together, and 4) nonsupplemented control. All treatments were administered from SD 0 to 28, followed by a 16-d wash-out period for a total study duration of 45 d and assayed to assess adult fly emergence, as well as fecal pH and fecal bacterial and total protein. A treatment × day interaction (P < 0.01) was observed for adult fly emergence. Fly emergence was not affected (P ≥ 0.05) by monensin, but was reduced during the 28 d treatment period when diflubenzuron was present (P = 0.01). Fecal pH, fecal bacterial protein, and total protein were not affected by any of the treatments (P ≥ 0.13). It is concluded that diflubenzuron is effective against both horn and house fly development regardless of the presence of monensin.

Effects of lipopolysaccharide on beta-catenin, aromatase, and estrogen production in bovine granulosa cells in vivo and in vitro.

Diseases resulting from Gram-negative bacterial infection can induce an immune response by releasing a lipopolysaccharide (LPS) endotoxin that may lead to impaired fertility in cows. To evaluate the effects of LPS on follicular dynamics in a subacute inflammatory disease state, 14 Angus heifers (BW = 413 kg±14) were blocked by weight and assigned to vehicle (n = 7) or LPS treated (n = 7) groups. Heifers received subcutaneous injections of saline (CON) or 2.0 µg/kg LPS on d 2, 5, and 8 of a select synch plus controlled internal drug release device (CIDR) follicular wave synchronization protocol. Fifty hours following CIDR withdrawal, ovaries were harvested, and follicular fluid was collected for hormone and LPS analysis. Daily blood samples were collected from d 0 to d 7. Beginning on d 8 blood samples were collected at 0, 16, 24, 32, 40, and 50 h following LPS challenge. Rectal temperatures were recorded prior to treatment and at regular intervals after each LPS challenge. Heifers treated with LPS exhibited mild (+0.5 °C) hyperthermia (P < 0.05) at 3, 4, and 8 h after the initial LPS challenge (d 2) when compared to vehicle-treated controls. Follicular fluid concentrations of estradiol (E2) increased (P = 0.04) in LPS-treated heifers compared to controls (1,595 ng/mL and 808 ng/mL±240, respectively), while follicular fluid progesterone (P4) concentrations did not differ (P = 0.27) between treatment groups. Additionally, LPS concentrations tended to be increased (P = 0.59) in dominant follicles of LPS-treated heifers, but no difference was detected (P = 0.81) in small developing follicles. To further delineate the impact of LPS on ovarian signaling pathways, a granulosa cell line (KGN) was incubated in the presence or absence of LPS (10 µg/mL) for 48 h. Cells were then collected for gene expression and protein analysis. Cells in both treatment groups expressed toll-like receptor 4, myeloid differentiation factor-2 receptor, and CD-14 complex genes required for LPS signaling. Cells treated with LPS exhibited decreased mRNA expression of aromatase (P = 0.03) and beta-catenin (P = 0.02). However, no change (P > 0.10) was detected in abundance of total beta-catenin protein or beta-catenin phosphorylated isoforms at serine 552 or 675. Based on results from this in vivo experiment, these investigators concluded that low doses of LPS can alter E2 concentrations and this effect may be modulated in part through beta-catenin regulation of aromatase transcription.

The Effect of Horn Fly (Diptera: Muscidae) Infestation on Behavior, Water, and Feed Intake, and Digestion Characteristics of Beef Cattle.

Seven mature Angus × Hereford crossbreed cows (544 ± 23.2 kg of body weight) fitted with duodenal and ruminal cannulas were used in a crossover design to evaluate the effects of horn fly (Haematobia irritans) (L.) infestation on horn fly-induced defensive behaviors, feed and water intake, as well as digestion characteristics of beef cows. Cows were randomly assigned to one of two treatment groups: 1) a horn fly infested (1,000 flies per cow) or 2) a noninfested control group. All cows were housed in individual screened pens. Fly counts and infestations were performed daily to maintain consistent populations throughout the study. Sudan hay (11.4 % crude protein, dry matter basis) was selected to mimic forage quality that range cattle frequently graze in the southwest region. Horn fly-induced host defensive behaviors were greater (P ≤ 0.0002) for horn fly infested cattle. Intake of water, dry matter, organic matter, crude protein, and neutral detergent fiber were similar (P ≥ 0.4765) among infested and noninfested cows. However, total volatile fatty acid concentration tended to increase (P = 0.0967) when horn flies were present. Dry matter fecal excretion and total tract digestion were not affected (P ≥ 0.2602) by horn fly infestation. These data indicate that horn fly infestations do not affect forage intake or digestion characteristics of beef cattle (Bos taurus) in confinement settings.

Crude glycerin alters beef cow ruminal fluid degradability of nutrients in vitro.

Crude glycerin (GLY) is used as an energy supplement for cattle and has been shown to increase propionate and decrease acetate concentrations in the rumen. Glycerin also decreases ruminal NH3, and therefore, we hypothesized that GLY in rumen fluid would decrease protein degradability of feedstuffs. The objective was to evaluate the effect of GLY in bovine rumen fluid on extent of protein degradability of alfalfa hay (ALF), dried distiller's grains (DDG), and soybean meal (SBM). The study was conducted using an in vitro incubator with four incubation jars. Each incubation jar contained 18 filter bags with 0.5 g of ground feed sample (six bags contained ALF, six contained DDG, and six contained SBM), six blank bags, 1,600 mL of buffer solution, 400-mL rumen fluid (collected from two ruminally cannulated cows), and one of the two treatments. Treatments were 12.7 g of deionized water (-GLY) or 12.7 g of GLY (+GLY) added to the 2-liter contents of each jar. Incubation jars were purged with CO2 and allowed to incubate at 39°C for 48 h. The 48-h incubation period was repeated for a total of three runs (the study was a randomized complete block design). Fluid samples from incubation jars were collected at 0 and 48 h into 25-mL vials for measurement of pH and analysis of VFA and NH3. At the end of the 48-h incubation period, filter bags containing feed samples were collected for determination of DM, NDF, and CP disappearance. The pH and NH3 concentrations of the contents of jars were lower (P ≤ 0.01) and total VFA concentrations were greater (P < 0.01) for +GLY than -GLY after 48 h of incubation. Molar percentages of acetate, isobutyrate, and isovalerate were lower (P < 0.01) and molar percentages of propionate and valerate were greater (P ≤ 0.02), resulting in a lower (P < 0.01) acetate-to-propionate ratio for +GLY compared with -GLY after the 48-h incubation period. A GLY × feed interaction (P < 0.01) occurred for DM and CP disappearance, where +GLY decreased DM and CP disappearance of SBM more than that of ALF and DDG. Disappearance of NDF from ALF, DDG, and SBM was lower for +GLY compared with -GLY (GLY; P < 0.01). These results suggest that supplementing glycerin to ruminants may cause an increase in RUP fraction escaping to the small intestine for potential digestion and absorption.

Nutritional recommendations of feedlot consulting nutritionists: The 2015 New Mexico State and Texas Tech University survey.

The 2015 feedlot consulting nutritionist survey is a collaborative project between New Mexico State University and Texas Tech University that focuses on summarizing the professional practices of consulting feedlot nutritionists and updates a 2007 survey. Forty-nine consulting feedlot nutritionists were asked to participate, of which 24 completed the survey. The nutritionists surveyed service over 14,000,000 cattle annually and were representatives from individual consulting practices (54.2%), corporate cattle feeding companies (20.8%), corporate feed manufacturing companies (20.8%), or a combination of consulting practices (4.2%). The survey was completed using a web-based survey tool and contained 101 questions that were divided into sections regarding general information about the consulting practice; general cattle management; receiving cattle management, diet adaption; mixers, feed mills, and feeding management; grains and grain processing; grain by-product use; roughage use; information about supplements and microingredients; liquid feed use; nutrient formulation; feed additive use; and information used as a basis for nutritional recommendations. In most cases, the results of the current survey were similar to those reported for the 2007 survey, with a few notable exceptions such as shifts in cattle numbers and preferences for specific feedstuffs. The present study introduced a number of new questions not included in the 2007 survey that focused on management strategies used in the receiving period. Data from this survey provide insight into current nutritional and management practices of consulting nutritionists and, as in past surveys, should be useful for informing national committees that make nutritional recommendations for cattle, as well as nutrition and management strategies employed within university research settings.

Effects of dietary urea concentration and zilpaterol hydrochloride on performance and carcass characteristics of finishing steers.

Cattle receiving zilpaterol hydrochloride () may recycle less N and require a greater supply of RDP. This study evaluated effects of ZH on performance and carcass characteristics of steers fed diets with increasing dietary RDP concentrations supplied as urea. Steers (429 animals; BW = 423 ± 4.5 kg) were sorted into 3 blocks according to BW and assigned to 1 of 6 treatments (6 pens per treatment) in a randomized complete block design. Treatments were a 2 × 3 factorial arrangement of either no ZH or ZH (75 mg ZH per steer daily) supplemented to finishing diets containing 0, 0.5, or 1.0% urea of dietary DM. Pen weights were recorded before treatment initiation; urea was fed for 27 d, and ZH treatments were fed for 24 d with a 3-d withdrawal period. Pen weights were recorded before transporting steers to a commercial abattoir. Continuous response variables were analyzed using the MIXED procedure and categorical data were analyzed using the GLIMMIX procedure of SAS. No ZH × dietary urea interactions ( ≥ 0.14) occurred for all performance and carcass response variables. Feeding ZH for the last 27 d (included a 3-d withdrawal period) of the finishing period increased ( < 0.01) ADG, decreased ( < 0.01) DMI, and increased ( < 0.01) G:F compared with no ZH. In addition, ZH increased HCW ( < 0.01), dressing percentage ( < 0.01), LM area ( < 0.01), and decreased ( = 0.01) yield grade. Increasing dietary urea linearly decreased ( = 0.01) ADG and DMI. A tendency for a linear decrease ( = 0.10) in HCW, and a tendency for a quadratic increase ( = 0.07) in marbling score were observed as urea increased in the diet. Results indicate that cattle supplemented with ZH do not require additional RDP in the diet, and that performance and carcass characteristics were negatively affected when urea was increased in the diet.

Beef Species Symposium: an assessment of the 1996 Beef NRC: metabolizable protein supply and demand and effectiveness of model performance prediction of beef females within extensive grazing systems.

Interannual variation of forage quantity and quality driven by precipitation events influence beef livestock production systems within the Southern and Northern Plains and Pacific West, which combined represent 60% (approximately 17.5 million) of the total beef cows in the United States. The beef cattle requirements published by the NRC are an important tool and excellent resource for both professionals and producers to use when implementing feeding practices and nutritional programs within the various production systems. The objectives of this paper include evaluation of the 1996 Beef NRC model in terms of effectiveness in predicting extensive range beef cow performance within arid and semiarid environments using available data sets, identifying model inefficiencies that could be refined to improve the precision of predicting protein supply and demand for range beef cows, and last, providing recommendations for future areas of research. An important addition to the current Beef NRC model would be to allow users to provide region-specific forage characteristics and the ability to describe supplement composition, amount, and delivery frequency. Beef NRC models would then need to be modified to account for the N recycling that occurs throughout a supplementation interval and the impact that this would have on microbial efficiency and microbial protein supply. The Beef NRC should also consider the role of ruminal and postruminal supply and demand of specific limiting AA. Additional considerations should include the partitioning effects of nitrogenous compounds under different physiological production stages (e.g., lactation, pregnancy, and periods of BW loss). The intent of information provided is to aid revision of the Beef NRC by providing supporting material for changes and identifying gaps in existing scientific literature where future research is needed to enhance the predictive precision and application of the Beef NRC models.

Effects of bacterial lipopolysaccharide injection on white blood cell counts, hematological variables, and serum glucose, insulin, and cortisol concentrations in ewes fed low- or high-protein diets.

Bacterial lipopolysaccharide endotoxins (LPS) elicit inflammatory responses reflective of acute bacterial infection. We determined if feeding ewes high-CP (15.5%) or low-CP (8.5%) diets for 10 d altered inflammatory responses to an intravenous bolus of 0 (control), 0.75 (L75), or 1.50 (L150) µg of LPS/kg of BW in a 2 × 3 factorial arrangement of treatments (n = 5/treatment). Rectal temperatures, heart and respiratory rates, blood leukocyte concentrations, and serum cortisol, insulin, and glucose concentrations were measured for 24 h after an LPS bolus (bolus = 0 h). In general, rectal temperatures were greater (P ≤ 0.05) in control ewes fed high CP, but LPS increased (P ≤ 0.05) rectal temperatures in a dose-dependent manner at most times between 2 and 24 h after the bolus. Peak rectal temperatures in L75 and L150 occurred 4 h after the bolus. A monophasic, dose-independent increase (P ≤ 0.023) in serum cortisol occurred from 0.5 to 24 h after the bolus, with peak cortisol at 4 h. Serum insulin was increased (P ≤ 0.016) by LPS in a dose-dependent manner from 4 to 24 h after the bolus. Insulin did not differ between control ewes fed high- and low-CP diets but was greater (P < 0.001) in L75 ewes fed low CP compared with high CP and in L150 ewes fed high CP compared with low CP. Increased insulin was not preceded by increased serum glucose. Total white blood cell concentrations were not affected (P ≥ 0.135) by LPS, but the neutrophil and monocyte fractions of white blood cells were increased (P ≤ 0.047) by LPS at 12 and 24 h and at 24 h after the bolus, respectively, and the lymphocyte fraction was increased (P = 0.037) at 2 h and decreased (P ≤ 0.006) at 12 and 24 h after the bolus. Red blood cell and hemoglobin concentrations and hematocrit (%) were increased (P ≤ 0.022) by LPS at 2 and 4 h after the bolus. Rectal temperatures and serum glucose were greater (P ≤ 0.033) in ewes fed a high-CP diet before LPS injection, but these effects were lost at and within 2.5 h of the bolus, respectively. Feeding high-CP diets for 10 d did not reduce inflammation in ewes during the first 24 h after LPS exposure but may benefit livestock by preventing acute insulin resistance when endotoxin exposure is mild.

Technical note: Effects of rumen passage on fluoxetine bioavailability in serum and effects of fluoxetine on serum prolactin concentration and demeanor in ewes.

Sheep are becoming increasingly important in medical research. The objective of the present study was to identify changes in bioactivity of fluoxetine during ruminal passage in ewes, and to examine the effects of fluoxetine administration on demeanor and serum prolactin concentration. Twelve mature ewes were administered saline (control), daily oral fluoxetine (40 mg), or alternate-day oral fluoxetine for 10 d. Four additional ewes were fitted with rumen cannulas and administered daily fluoxetine by abomasal deposition. Serum samples were collected daily for 15 d. Serum fluoxetine concentrations (ELISA) were greater (P < 0.001) in ewes in all fluoxetine treatments compared with controls on d 2. Serum fluoxetine concentrations in ewes receiving daily abomasal dosages were greater (P < 0.007) than those in controls on d 2 to 12 and were greater than those in ewes receiving daily or alternate-day oral fluoxetine on d 3 to 12. Serum prolactin concentration (RIA) did not differ (P = 0.137) among treatments and was only weakly correlated with serum fluoxetine concentration (r = 0.20, P = 0.041), and regression analysis revealed that very little variation in serum prolactin concentration was due to serum fluoxetine concentration (R(2) = 0.04, P = 0.082). Demeanor ratings on d 1 to 12 remained at normal in all treatment groups (P > 0.362). However, in ewes that had received an abomasal dosage of fluoxetine, demeanor scores decreased (P < 0.029) on d 13 and 14 before returning to normal on d 15 (P = 0.397). This study indicates that mature ewes may provide a suitable model for the study of fluoxetine, but that a larger oral dosage may be required relative to the human dosage to overcome partial loss of bioactivity during ruminal passage.

Effects of dietary protein and bacterial lipopolysaccharide infusion on nitrogen metabolism and hormonal responses of growing beef steers.

Purified lipopolysaccharide (LPS) infusion in cattle induces clinical and metabolic responses similar to gram-negative bacterial infection. Effects of LPS and dietary protein on rectal temperature, serum hormones, haptoglobin, plasma urea N and AA, and N balance were evaluated in 24 steers (250 +/- 2.8 kg of BW). Treatments were a 2 x 3 factorial of LPS (0 vs. 1.5 microg/kg of BW; -LPS vs. +LPS) and diets containing (DM basis) 1) 14.5% CP, 11.6% ruminally degradable protein (RDP), and 2.9% ruminally undegradable protein (RUP; CP14.5CON); 2) 16.3% CP, 13.4% RDP, and 2.9% RUP (CP16RDP); and 3) 16.1% CP, 11.2% RDP, and 4.9% RUP (CP16RUP). Diet RDP and RUP were altered using casein, fish meal, and corn gluten meal. Steers were adapted to diets (1.1 Mcal/kg of NE(g); DM fed at 1.8% BW) for 14 d and were infused (intravenously 1 mL/min) with LPS (in 100 mL of saline) on d 15. Rectal temperature and serum cortisol, prolactin, haptoglobin, and insulin increased, glucose initially increased and then declined, and serum thyroxine and triiodothyronine decreased for +LPS vs. -LPS steers (LPS x hour; P < 0.01). Serum IGF-I was less (P < 0.01) for +LPS vs. -LPS steers. Plasma urea N increased in response to LPS (LPS x hour; P = 0.02) and was greater for +LPS steers fed CP16RDP and CP16RUP vs. CP14.5CON, but greater in -LPS steers fed CP16RUP vs. CP16RDP and CP14.5CON (LPS x diet; P = 0.04). Plasma Met, Thr, Leu, Ile, Phe, Trp, Gly, Ser, Asn, and Tyr decreased, and plasma Ala increased in response to LPS (LPS x hour; P < 0.01). Plasma Orn initially increased and then decreased in +LPS vs. -LPS steers (LPS x hour; P < 0.01). No LPS x diet interactions (P > or = 0.15) occurred for DM, OM, NDF and N intake, fecal excretion, or apparent digestibility. Dietary DM, OM, NDF, and N intake, and retained N were less (P < 0.01) for +LPS than -LPS steers. Total N intake, apparent N digestibility, and retained N were greater (P < or = 0.05) for steers fed CP16RDP and CP16RUP vs. CP14.5CON. An LPS x diet interaction (P = 0.05) occurred for N retention (% N intake) because N retention was less for +LPS than -LPS steers when fed CP14.5CON, but not different between +LPS and -LPS steers when fed CP16RDP and CP16RUP. These results demonstrate that LPS infusion alters serum hormones, plasma AA, and N balance in cattle and imply that growing steers exposed to LPS may require greater dietary protein concentrations to account for altered intake and metabolic AA demand.

Effects of rumen-protected methionine supplementation and bacterial lipopolysaccharide infusion on nitrogen metabolism and hormonal responses of growing beef steers.

Metabolic demand for sulfur-containing AA increases during inflammation in nonruminants. Therefore, Met supplementation may alleviate the negative effects of infection on N balance. Effects of gram-negative bacterial lipopolysaccharide (LPS) and supplemental dietary Met on N balance, serum hormones and haptoglobin, and plasma urea-N and AA were evaluated in 20 Angus-cross steers (BW = 262 +/- 6.3 kg). Treatments (2 x 2 factorial) were infusion of no LPS (-LPS) or a prolonged low dose of LPS (+LPS) and dietary supplementation of no (-MET) or 14 g/d (+MET) of rumen-protected Met (providing 7.9 g/d of dl-Met). Steers were adapted to a roughage-based diet (DMI = 1.4% of BW daily) and supplemental Met for 14 d, and were then infused (1 mL/min via intravenous catheter) with LPS on d 1 (2 microg/kg of BW) and 3 (1 microg/kg of BW) of a 5-d collection period. Blood was collected on d 1, before LPS infusion, and at 2, 4, 6, 8, 10, 12, and 24 h after LPS challenge. Diet samples, feed refusals, feces, and urine were collected daily for 5 d. Rectal temperature and serum concentrations of cortisol, prolactin, tumor necrosis factor-alpha, and haptoglobin increased, whereas thyroxine and triiodothyronine decreased for +LPS vs. -LPS steers (LPS x h; P < 0.01). Plasma urea-N was greater for +LPS than -LPS steers (LPS; P = 0.03), and serum IGF-1 was not affected (P > or = 0.26) by LPS or Met. Plasma concentrations of Thr, Lys, Leu, Ile, Phe, Trp, Asn, Glu, and Orn decreased, plasma Ala increased, and Gly and Ser initially increased, then declined in +LPS vs. -LPS steers (LPS x h; P < or = 0.04). Plasma Met was greater for +MET than -MET steers before LPS infusion, but declined in +MET steers after LPS infusion (LPS x Met x h; P < 0.01). By design, DMI was not different, but DM digested was less (P = 0.04) for +LPS than -LPS steers. Infusion of LPS did not affect (P > or = 0.24) N intake, fecal N excretion, or N digested, but resulted in greater (P < 0.01) urinary N excretion and less (P < 0.01) N retention. The absence of an LPS x Met interaction (P = 0.26) for N retention indicates that supplemental Met does not improve the N utilization of growing beef steers exposed to a gram-negative bacterial endotoxin. Decreases in plasma concentrations of several essential AA in +LPS steers suggest that metabolic demand for these AA likely increased in steers exposed to endotoxin.

Limiting amino acids for growing lambs fed a diet low in ruminally undegradable protein.

Ruminally cannulated Rambouillet wether lambs were used in three 6 x 6 Latin square experiments (n = 6/experiment) to determine which essential AA limit N retention. Lambs (BW = 36.9 +/- 1.9 kg for Exp. 1, 35.1 +/- 1.4 kg for Exp. 2, and 46.0 +/- 1.3 kg for Exp. 3) were housed in metabolism crates and limit-fed (DMI = approx. 1.8% of BW daily) twice daily a soybean hull-based diet low in ruminally undegradable protein. Treatments for Exp. 1 were continuous abomasal infusions of a solution (500 mL/d) containing 1) no AA (CON), 2) a mixture of 10 essential AA and 2 nonessential AA (10EAA), 3) 10EAA with Met removed, 4) 10EAA with Lys removed, 5) 10EAA with His removed, and 6) 10EAA with Thr removed. Treatments for Exp. 2 were abomasal infusions of 1) CON, 2) 10EAA, 3) 10EAA with Leu, Ile, and Val removed (-BCAA), 4) 10EAA with Arg removed, 5) 10EAA with Phe removed, and 6) 10EAA with Trp removed. Treatments for Exp. 3 were abomasal infusions of 1) CON, 2) 10EAA, 3) -BCAA, 4) 10EAA with Leu removed, 5) 10EAA with Ile removed, and 6) 10EAA with Val removed. All lambs received continuous infusions of acetate and propionate into the rumen and dextrose into the abomasum to supply additional energy. Periods were 7 d: 3 d for adaptation to abomasally infused treatments and 4 d for fecal and urinary collections. Blood samples were collected 3 h after feeding on d 7. In all 3 experiments, N retention was greater (P < 0.10) for lambs receiving 10EAA vs. CON, demonstrating that the basal AA supply from CON was limiting. Removal of each of the essential AA from 10EAA decreased (P < 0.10) their concentrations in plasma (except for Trp), indicating that 10EAA supplied these AA in excess of the animal's requirement. In Exp. 1, N retention (g/d) decreased (P < 0.10) in response to the removal of Met and Thr, but was not affected by removal of Lys and His from 10EAA. In Exp. 2, N retention decreased (P < 0.10) in response to removal of all 3 branched-chain AA, Arg, and Trp, whereas the removal of Phe from 10EAA did not affect N retention. In Exp. 3, N retention decreased (P < 0.10) in response to removal of branched-chain AA and Val, but was not affected by the omission of Leu and Ile from 10EAA. The results of this research demonstrated that Met, Thr, Arg, Trp, and Val limited N retention of lambs fed a diet low in ruminally undegradable protein.

Intrauterine bacterial inoculation and level of dietary methionine alter amino acid metabolism in nulliparous yearling ewes.

Using an intrauterine bacterial inoculation method, our objective was to determine the effects of acute sepsis and level of dietary metabolizable Met (MM) on splanchnic metabolism of AA in ewes. Twenty-four nulliparous yearling Rambouillet-cross ewes (initial BW = 65.1 +/- 0.6 kg), surgically fitted with chronic-indwelling catheters in hepatic and portal veins, a mesenteric vein and artery, and the uterine lumen, were assigned to a 2 x 2 factorial arrangement of treatments. Factors were intrauterine bacterial inoculation (noninoculated vs. inoculated) and level of MM [low (2.28 g/d) vs. high (3.91 g/d)]. Beginning 12 h before sampling, inoculated and noninoculated ewes received 10-mL intrauterine infusions of Escherichia coli (9.69 x 10(11) cfu) + Arcanobacterium pyogenes (2.76 x 10(12) cfu) and of sterile saline, respectively. Uterine infection was induced in ewes that received intrauterine bacterial inoculations, but not in ewes infused with sterile saline. Bacterial inoculation resulted in increased hepatic release and plasma concentrations of aromatic AA used for acute-phase protein synthesis, increased hepatic removal and decreased plasma concentrations of AA used for glutathione synthesis, and decreased plasma concentrations of some gluconeogenic and acetogenic AA used for glucose recycling and anaerobic energy production, respectively (P < 0.05). In ewes fed high-MM diets, compared with low-MM diets, a consistent net hepatic uptake of Phe occurred throughout the sampling period, more Asp was released from the portal-drained viscera, and hepatic vein glucose concentrations were greater (P < 0.05). We conclude that Met seemed to be limiting in low-MM ewes, and as such, would continue to be limiting during sepsis. However, additional MM, in excess of the dietary requirement, would not necessarily result in a benefit to ewes experiencing acute sepsis.

Supplemental methionine and urea for gestating beef cows consuming low quality forage diets.

A study was conducted to evaluate Met requirements of late-gestation beef cows consuming low quality forages on the premise that inadequate supply of metabolizable AA may limit protein accretion during pregnancy. Five ruminally cannulated, multiparous late-gestation beef cows (490 +/- 27 kg), of predominantly Angus (> or =75%) with Hereford and Simmental breeding, were used in a 5 x 5 Latin square experiment to evaluate the effects of postruminal dl-Met supplementation on N retention, serum metabolites, and plasma AA concentrations during the third trimester of pregnancy. The basal diet was fed individually, and weights of refusals were recorded for N intake determination. Treatments consisted of no urea, urea (0.053 +/- 0.002 g/kg of BW daily), urea + 5 g of Met/d, urea + 10 g of Met/d, and urea + 15 g of Met/d. Cows were adapted to the experimental diet 30 d before the beginning of the study, with periods lasting for 14 d; 4 d to allow for clearance of the previous treatment effects, 4 d for adaptation to the treatments, and 6 d for total fecal and urine collection. Blood samples were collected every 4 h on d 13 of each period for analysis of serum metabolites and plasma AA. Inclusion of urea increased DM and OM intakes (urea vs. no urea; P = 0.05), but no further improvement in intake was observed with inclusion of Met. Serum urea concentrations increased with inclusion of urea (P = 0.03) and responded quadratically (P = 0.06) when Met was added, with the lowest concentration observed in the urea + 5 g of Met/d treatment. More N was retained with the inclusion of urea (P = 0.04), and N retention increased linearly (P = 0.07) with inclusion of Met. Plasma Met concentration increased linearly (P < 0.01) with inclusion of Met. These data suggest that Met was a limiting AA and that supplementation of a combination of urea and 5 g/d of rumen-protected Met to low quality, forage diets will improve N retention and promote protein accretion during late pregnancy.

Effect of glycine and vitamin supplementation on sulphur amino acid utilization by growing cattle.

Effects of glycine (Gly) and B-vitamins on sulphur amino acid (AA) utilization were studied in growing steers maintained under conditions where methionine (Met) was first limiting. Conditions were generated by limit feeding a diet low in ruminally non-degraded protein and abomasally infusing an AA mixture limiting in Met. Retained N tended (p = 0.07) to improve when steers received 10 mg folate, 10 mg vitamin B6, and 0.10 mg vitamin B12 daily. Hepatic vitamin B12 (p = 0.08) and folate (p = 0.05) concentrations increased with vitamin supplementation. In another trial, factorial treatments were 2 or 5 g/day L-Met and 0 or 50 g/day Gly infused abomasally. Retained N increased (p < 0.05) in response to Met, and responses were numerically larger in the presence of supplemental Gly. In a different trial, factorial treatments were 0 or 2.4 g/day L-cysteine (Cys) and 0 or 40 g/day Gly. Retained N was not affected by Cys in the absence of Gly, but was increased by Cys when Gly was supplemented (interaction, p = 0.01). B-vitamin status may affect sparing of Met by Cys. Supplemental Gly improved responses to supplemental Met and Cys.

Methionine as a methyl group donor in growing cattle.

Holstein steers were used in two 5 x 5 Latin square experiments to evaluate the sparing of methionine by alternative sources of methyl groups (betaine and choline). Steers were housed in metabolism crates and limit-fed a soybean hull-based diet high in rumen degradable protein. To increase energy supply, ruminal infusions of volatile fatty acids and abomasal infusions of glucose were provided. An amino acid mixture, limiting in methionine, was infused abomasally to ensure that nonsulfur amino acids did not limit protein synthesis. Treatments for Exp. 1 were abomasal infusion of 1) water, 2) 2 g/d L-methionine, 3) 1.7 g/d L-cysteine, 4) 1.6 g/d betaine, and 5) 1.7 g/d L-cysteine + 1.6 g/d betaine. Treatments for Exp. 2 were abomasal infusion of 1) water, 2) 2 g/d L-methionine, 3) 8 g/d betaine, 4) 16 g/d betaine, and 5) 8 g/d choline. In both experiments, nitrogen retention increased in response to methionine (P < 0.05), demonstrating a deficiency of sulfur amino acids. Responses to cysteine, betaine, and choline were all small and not significant. The lack of response to cysteine indicates that the response to methionine was not due to transsulfuration to cysteine or that cysteine supply did not alter the flux of methionine through transsulfuration. The lack of response to betaine suggests that the steers' needs for methyl groups were met by the dietary conditions or that betaine was relatively inefficient in increasing the remethylation of homocysteine to methionine and, thereby, reducing the synthesis of cysteine from homocysteine. Under our experimental conditions, responses to methionine were likely due to a correction of a deficiency of methionine per se rather than of methyl group donors.

Supplemental betaine and peroxide-treated feather meal for finishing cattle.

These studies evaluated the effects of betaine, provided either as feed-grade betaine or as concentrated separator by-product (CSB; desugared beet molasses), on performance and carcass characteristics of finishing cattle. In Exp. 1, 175 steers (410 kg initial BW) were fed a finishing diet based on steam-flaked and dry-rolled corn, and treatments included 10.5 and 21 g/d feed-grade betaine and 250 and 500 g/d CSB (supplying 15.5 and 31 g/d of betaine, respectively). Steers fed feed-grade betaine had greater (linear and quadratic effects, P < 0.1) DMI than control steers, but ADG and gain efficiencies were not affected by treatment. Dressing percent and backfat thickness was greater (P < 0.1) for steers that received feed-grade betaine than for controls. Longissimus muscle area was lower (P < 0.1) for steers supplemented with either feed-grade betaine or CSB than for control steers. Yield grades were higher for cattle receiving feed-grade betaine (quadratic effect, P < 0.1) than for control steers. Marbling scores were not affected by supplemental betaine, but the percentage of carcasses grading USDA Select was lower (linear and quadratic effects, P < 0.1) for steers fed feed-grade betaine than for control steers, predominantly due to a greater percentage grading USDA Choice. In Exp. 2, 312 heifers (343 kg initial BW) were used in a finishing study to evaluate the effects of graded levels of feed-grade betaine and peroxide-treated feather meal on performance and carcass characteristics. Treatments included two finishing diets (containing peroxide-treated or untreated feather meal) and four levels (0, 4, 8, and 12 g/d) of feed-grade betaine arranged in a 2 x 4 factorial. No significant interactions occurred between treatment of feather meal and betaine. Treatment of feather meal with hydrogen peroxide (5% wt/wt) increased in situ protein degradability but did not alter DMI, ADG, gain efficiencies, or carcass characteristics of heifers when it replaced untreated feather meal in the diet. Top-dressing feed-grade betaine to the diets had no effect on DMI, ADG, and gain efficiencies. Marbling scores were greater (cubic effect, P < 0.05) for heifers fed diets top-dressed with 4 and 12 g/d of feed-grade betaine, but other carcass characteristics were not altered significantly. Overall, feed-grade betaine and CSB did not alter growth performance, but did have minor effects on carcass characteristics.

Branched-chain amino acids for growing cattle limit-fed soybean hull-based diets.

Five ruminally cannulated Holstein steers (176 kg) were used in a 5 x 5 Latin square to evaluate the effects of branched-chain AA supplementation on N retention and plasma AA concentrations of steers. Steers were limit-fed (3.0 kg/d of DM) twice daily diets low in ruminally undegradable protein (72% soybean hulls, 19% alfalfa, 5% molasses, and 4% vitamins and minerals). Acetate (400 g/d) was continuously infused into the rumen. Treatments were continuous abomasal infusions of 1) 115 g/d of a mixture of 10 essential AA designed to exceed the steers' requirements (10AA), 2) 10AA with Leu removed, 3) 10AA with Ile removed, 4) 10AA with Val removed, and 5) 10AA with all three branched-chain AA removed. Experimental periods were 7 d, with 3 d for adaptation to treatments and 4 d for total fecal and urinary collections for N balance. Blood samples were collected 5 h after feeding on d 7. Retained N decreased in response to removal of Leu (P < 0.06), Val (P < 0.05), or all three branched-chain AA (P < 0.05). Plasma Leu concentrations decreased (P < 0.05) in response to removal of Leu and all three branched-chain AA. Plasma Ile concentrations decreased (P < 0.05) in response to removal of Ile and all three branched-chain AA but increased (P < 0.05) in response to removal of Leu. Plasma Val concentrations decreased (P < 0.05) in response to removal of Val and all three branched-chain AA but increased (P < 0.05) in response to removal of Leu. Responses in N balance and plasma AA concentrations of growing cattle limit-fed soybean hull-based diets demonstrate limitations in the basal supply of Leu and Val but not Ile provided that supplies of all other essential AA are met.

Effects of supplemental alfalfa hay on the digestion of soybean hull-based diets by cattle.

We evaluated the optimal level of alfalfa inclusion in soybean hull-based diets. In Exp. 1, 20 Holstein steers (319 kg of BW) were used in a complete block design. Treatments included a soybean hull mix (95.7% soybean hulls, 3% molasses, 0.5% urea, 0.8% mineral mix; DM basis) fed alone (100:0) or with 10.4, 20.7, or 30.9% (DM basis) coarsely chopped alfalfa hay (90:10, 80:20, and 70:30, respectively) or alfalfa alone (0:100). Diets were fed once daily at 1.75% (DM basis) of BW. In some cases, orts were present, which caused DM, OM, and NDF intakes to decrease (linear, P < 0.05) as alfalfa was added to the diets. Digestibilities of DM, OM, and NDF decreased linearly (P < 0.05) as alfalfa was added to the diets, but quadratic responses (P < 0.05) indicated that positive associative effects occurred between soybean hulls and alfalfa. Liquid dilution rates increased (linear, P < 0.05) with alfalfa additions to the diets and also demonstrated positive associative effects between soybean hulls and alfalfa (quadratic, P < 0.05). Solid passage rates were similar for 100:0 and 0:100 but were increased (quadratically and cubically, P < 0.05) when combinations of soybean hulls and alfalfa were fed. In Exp. 2, in vitro NDF digestibilities were measured for soybean hulls, alfalfa, and a blend of 85% soybean hulls and 15% alfalfa, each with no N source or supplemented with casein or urea to ascertain the effects of protein from alfalfa on digestibility. Disappearances were increased (P < 0.05) by addition of urea or casein, but no interactions between substrate and N supplement were observed. Addition of 30% alfalfa to diets consisting primarily of soybean hulls led to positive associative effects on diet digestibility, but alfalfa additions led to increased liquid and solid passage rates, suggesting that the benefit was not a result of slower passage of soybean hulls from the rumen.