Clearance of para-aminohippuric acid in wethers consuming locoweed

AIM: To validate the use of para-aminohippuric acid (PAH) as a marker for measuring blood flow in wethers consuming a mixed diet of locoweed and blue grama hay. METHODS: Fourteen sheep, stratified by bodyweight (BW), were assigned to one of three treatments: 0.8 mg swainsonine (SW)/kg BW (HI), 0.2 mg SW/kg BW (LO), and no SW (Control). Sheep were fed various ratios of locoweed and blue grama hay to deliver SW treatments, for 28 days prior to infusion of PAH. Concentrations of SW and activities of alkaline phosphatase (Alk-P) and aspartate aminotransferase (AST) in serum were measured to confirm exposure to SW and subclinical intoxication. A single 20-ml injection of 5% PAH was delivered into the jugular vein after subclinical intoxication had been achieved. Blood samples were collected and serum analysed for PAH immediately prior to injection, then every 5 min from 5-30 min, and every 10 min from 30-60 min, following injection of PAH. RESULTS: Effective delivery of SW was evident from the greater concentrations of SW measured in the serum of HI compared with LO animals (p<0.05). No significant differences were detected in the rate of elimination (range 0.097-0.108 L/min), elimination half-life (range 6.62-7.24 min), apparent volume of distribution for the central compartment (range 7.14-9.72 L), and clearance (range 0.73-0.92 L/min) of PAH, between treatments. CONCLUSIONS: Subclinical intoxication with SW did not affect the pharmacokinetics of PAH. Thus, use of downstream dilution of PAH is a valid method to determine the rate of blood flow in nutrient flux experiments that involve consumption of locoweed.     

Toxicokinetic profile of swainsonine following exposure to locoweed (Oxytropis sericea) in naïve and previously exposed sheep

AIM: To determine the toxicokinetic profiles of swainsonine (SW) in sheep previously (subacute) and not previously (acute) exposed to locoweed. METHODS: Twenty-nine wethers were stratified by bodyweight (BW; 68.0 (SE 7.6) kg) and randomly assigned to one of six treatments. Treatments were: 0 (n=5), 0.4 (n=5), and 1.6 (n=5) mg SW/kg BW for Trial 1, and 0 (n=4), 0.2 (n=5), and 0.8 (n=5) mg SW/kg BW for Trial 2. Acute exposure in both trials included adaptation to blue grama (Bouteloua gracilis) hay for 14 days and no previous exposure to locoweed (i.e. SW), followed by administration of a single oral dose of SW prepared from an extract of locoweed, in the doses described above. Subacute exposure comprised ingestion of a blue grama and locoweed (428 microg SW/g locoweed) diet for 21 days in Trial 1 and 28 days in Trial 2, followed by removal from locoweed for 5 days, then an oral dose of SW, as above. Quantities of locoweed fed in the diet were adjusted to achieve the dose rates specified for each treatment. Blood samples were collected via jugular venepuncture twice daily for 3 days prior to initial exposure to SW and then every 7 days for the duration of the trials, to monitor serum alkaline phosphatase (Alk-P) and aspartate aminotransferase (AST) activities. For intensive sampling periods, SW was administered immediately following blood sampling at 0 h, and blood samples were collected at hourly intervals from 0-12 h, 3-h intervals from 15-24 h, 6-h intervals from 30-48 h, and 12-h intervals from 60-168 h. Concentrations of SW in serum and locoweed extract were determined using the alpha-mannosidase inhibition assay (detection limit=25 ng/ml). Rates of absorption and elimination of SW from serum were calculated for each animal, using exponential curve fits of the concentration of SW in serum concentration vs time plots. RESULTS: In both trials, SW was detected in serum in all animals exposed to locoweed. Elevated (p<0.05) serum Alk-P and AST activities indicated that subclinical SW intoxication was induced during the subacute exposure phase. Calculated rates of elimination were faster (p<0.001) for the 1.6 vs 0.4 (Trial 1) and 0.8 vs 0.2 (Trial 2) mg SW/kg BW doses. Rates of elimination indicated that, in both trials, SW was removed from serum faster (p<0.06) following acute exposure than subacute exposure. Higher exposure rates to SW resulted in higher concentrations of SW in serum within a trial. CONCLUSIONS: Multiple compartments were involved in the kinetics of SW, and dose and previous exposure altered the toxicokinetics of SW. CLININCAL RELEVANCE: Should the true elimination half-life prove to be as high or higher than the 95 h demonstrated for the treatment using 0.4 mg SW/kg BW in Trial 1, then withdrawal periods for clearing SW from sheep should be >40 days (assuming 10 half-lives to clear the compound).     

Early aspects of locoweed toxicosis and evaluation of a mineral supplement or clinoptilolite as dietary treatments.

Sixteen crossbred beef heifers were used to determine the efficacy of serum clinical profiles as diagnostic tools for detection of early stages of locoweed toxicity and to test the ability of two mineral supplements for prevention or therapy of toxicosis. Dietary treatments were (DM basis) 1) 100% sorghum sudangrass hay, 2) 80% sorghum sudangrass hay:20% locoweed, 3) 80% sorghum sudangrass hay:20% locoweed plus 100 g of Silent Herder Mineral Mix (a mineral supplement reported to alleviate locoweed toxicity), and 4) 80% sorghum sudangrass hay:20% locoweed plus 100 g of clinoptilolite (a natural zeolite clay). Diets were fed at 1.5% of BW for 28 d, after which heifers had ad libitum access to sorghum sudangrass hay for 14 d. Jugular blood was sampled before feeding every 7 d, and at 2, 4, 6, and 8 h after feeding on d 28. Compared with controls, heifers fed locoweed had elevated (P less than .01) serum alkaline phosphatase activities from d 7 through 35. On d 42, alkaline phosphatase activities in heifers previously fed locoweed were lower (P less than .05) than in control heifers. Serum glutamic oxaloacetic transaminase activities were elevated (P less than .01) in heifers fed locoweed from d 7 through 42 compared with control heifers. In heifers fed locoweed, serum Fe concentrations were less (P less than .01) on d 7 through 28, but no treatment effects were noted (P greater than .10) on d 35 or 42.(ABSTRACT TRUNCATED AT 250 WORDS)     

The interaction of harvesting time of day of switchgrass hay and ruminal degradability of supplemental protein offered to beef steers

The objective of this study was to evaluate an interaction between harvest at 0600 (AM) vs. 1800 (PM) with high (HI) or low (LO) ruminal degradability of a protein supplement to change voluntary intake, digestion, or N retention by steers offered switchgrass (Panicum virgatum L.) hay. Black steers (255 +/- 14 kg of BW) were blocked by BW, and then randomly assigned (5 steers each) to AM/HI, PM/HI, AM/LO, or PM/LO treatment groups. Steers were group-housed in covered, outdoor pens with individual feeding gates. After adaptation and standardization, intake was measured for 21 d followed by a digestion trial (5 d of total collection). Steers were offered 767 (LO) or 825 (HI) g/d of supplement to provide 268 g of CP/d. Compared with AM, PM had greater (P = 0.01) concentrations of total nonstructural carbohydrate (TNC, 71 vs. 56 g/kg of DM), and lesser concentrations of NDF (760 vs. 770 g/kg of DM, P = 0.02), ADF (417 vs. 427 g/kg of DM, P = 0.02), and CP (55.9 vs. 58.6 g/ kg of DM, P = 0.07). Protein fractions A, B(2), and B(3) were similar for AM and PM, but HI contained more (P < 0.02) A (694 vs. 296 g/kg of protein) and less B(2) (174 vs. 554 g/kg of protein) fraction than LO. Harvest interacted with supplement to increase (P = 0.07) ad libitum digestible DMI for steers offered PM/HI (11.4 g/kg of BW daily) compared with steers offered PM/LO (10.2 g/kg of BW daily), but there was no difference for steers offered AM/LO or AM/HI (10.7 g/kg of BW). Apparent digestibilities of DM (594 vs. 571 g/kg of intake), NDF (591 vs. 562 g/kg of intake), ADF (585 vs. 566 g/kg of intake), and N (651 vs. 632 g/kg of intake) were greater (P < 0.04) for PM than for AM. Apparent digestibility of N was greater (P = 0.02) for HI (652 g/ kg of intake) vs. LO (631 g/kg of intake). Interactions between harvest and supplement for apparent digestibilities of NDF (P = 0.09) and ADF (P = 0.03) were due to no change or an increase in digestibility in response to increased ruminal degradability of supplement in steers offered PM harvest, whereas increased ruminal degradability of supplement decreased digestibility of NDF and ADF in steers offered AM harvest. Treatments did not affect hay intake (3.93 kg/d), N retained (15.8 g/d), or plasma urea N (5.25 mM) during ad libitum intake. Greater TNC in PM vs. AM harvest was not sufficient by itself to increase total voluntary DMI, but greater protein degradability interacted with harvest time to increase ruminal fiber digestibility and digestible DMI of beef steers offered PM vs. AM harvest.     

Effects of an Estradiol Implant on Locoweed Consumption,Toxicity, and Recovery in Growing Beef Steers

Two experiments were conducted to determine effects of an estradiol implant (Compudose®) on locoweed consumption and toxicity in growing steers. In Exp. 1, 16 crossbred steers (185.3 ± 6.1 kg) were randomly assigned to two replicated treatments and received either an estradiol implant or no implant. Steers were assigned to one of four pastures and were rotated through all pastures, which differed in locoweed distribution, to allow equal access. Bite counts were recorded twice daily at 0600 and 1700 h during a period when steers were likely to consume locoweed. For bite counts, steers were observed for 5 min each, starting at 0600 and at 1700 h, and the number of bites taken of cool- and warm-season grasses, forbs, and locoweed were recorded. Blood was collected on d 0, 7, 28, 35, 63, and 119, and individual BW was recorded on d 0, 35, 63, and 119. Proportion of bites of locoweed consumed by implanted vs nonimplanted steers did not differ (P>0.10). Likewise, ADG, serum alkaline phosphatase activity, and thyroxine concentrations did not differ (P>0.10) between implanted and nonimplanted steers. In Exp. 2, 20 crossbred steers (212.3 ± 6.1 kg) were divided into four groups and individually fed in a 2 × 2 factorial arrangement of treatments. Treatments included: 1) estradiol implant + locoweed, 2) implant, no locoweed, 3) no implant + locoweed, and 4) no implant, no locoweed. Steers were implanted at d 0 and fed either a ground forage diet containing 80% sudangrass hay and 20% locoweed, or a diet of 100% sudangrass hay. Implanted steers had improved ADG vs nonimplanted steers (P<0.10) through 63 d on trial, but no differences were observed in steers fed locoweed vs sudangrass hay diets (P>0.90; locoweed x implant, P>0.10). Alkaline phosphatase activity was greater (P<0.05) for steers fed locoweed vs those not receiving locoweed on d 7, 14, and 21, whereas, thyroxine concentration was lower (P<0.06) in steers fed locoweed than those not fed locoweed on d 14 and 21. Estradiol concentrations were greater in implanted steers vs those not implanted (P<0.05). These results suggest no effect of an estrogen implant on locoweed consumption or on severity of locoweed toxicity by beef steers.     

Appearance and disappearance of swainsonine in serum and milk of lactating ruminants with nursing young following a single dose exposure to swainsonine (locoweed; Oxytropis sericea)

A series of experiments were conducted to investigate the elimination of swainsonine in the milk of lactating ruminants following a single dose oral exposure to swainsonine (locoweed; Oxytropis sericea) and to assess subsequent subclinical effects on the mothers and their nursing young. In a preliminary experiment, lactating ewes were gavaged with locoweed providing 0.8 mg swainsonine/kg BW (n = 4; BW = 75.8 +/- 3.6 kg; lactation = d 45) and lactating cows were offered up to 2.0 mg swainsonine/kg BW free choice (n = 16; BW = 389.6 +/- 20.9 kg; lactation = d 90). Serum and milk were collected at h 0 (before treatment), 3, 6, 12, and 24 for ewes, and h 0 (before treatment), 6, 12, 18, and 24 for cows. Swainsonine was highest (P < 0.05) by h 6 in the serum and milk of ewes. Consumption of at least 0.61 mg swainsonine/kg BW induced consistent (> 0.025 microg/mL) appearance of swainsonine in cow serum and milk. In response to the results obtained in the preliminary experiment, a subsequent experiment utilizing lactating ewes (n = 13; BW = 74.8 +/- 6.4 kg; lactation = d 30) and cows (n = 13; BW = 460.8 +/- 51.9 kg; lactation = d 90) was conducted. Each lactating ruminant was gavaged with a locoweed extract to provide 0 (control), 0.2, or 0.8 mg swainsonine/kg BW and individually penned with her nursing young. Serum and milk from the mothers and serum from the nursing young were collected at h 0 (before treatment), 3, 6, 9, 12, 24 and 48 (an additional sample was obtained at h 72 for ewes and lambs). Serum and milk swainsonine was higher (P < 0.05) in the 0.8 mg treated groups and maximal (P < 0.05) concentrations occurred from h 3 to 6 for ewes and h 6 to 12 h for cows (P < 0.05). Rises in alkaline phosphatase activity indicated subclinical toxicity in the treated ewes (P < 0.05). Following a single dose oral exposure to 0.2 and 0.8 mg swainsonine/kg BW provided by a locoweed extract, swainsonine was detected in the serum and milk of lactating ewes and cows, and rises in serum alkaline phosphatase activity were observed in the ewes. Neither swainsonine nor changes in alkaline phosphatase activity was detected in the serum of the lambs and calves nursing the ewes and cows dosed with swainsonine.     

Effect of cutting height and genetics on composition, intake, and digestibility of corn silage by beef heifers

This research was conducted to determine the effect of corn genetics and cutting height on the composition and nutritive characteristics of corn silage. An in situ study involving eight commercially available corn hybrids indicated main effects and interactions (P < 0.01) of hybrid and cutting height on NDF, ADF, and starch content and on in situ DM and NDF degradablility. Four ruminally cannulated Angus heifers (initial BW = 378 +/- 3 kg) were used in a 4 x 4 Latin square digestion experiment with a 2 x 2 factorial treatment arrangement. Main effects and interactions of hybrid (Pioneer Hi-Bred Int., Inc., hybrids 3335 and 3223) and cutting height (LO = 20.3 cm, and HI = 61 cm) were evaluated. Dietary treatment consisted of 40% chopped alfalfa hay and 60% corn silage. Although corn silage hybrids used were of equivalent maturity at harvest (60% milkline), 3335 treatments had 37.8% starch and 34.8% NDF, whereas 3223 treatments had 33.7% starch and 38.6% NDF. The LO treatments averaged 3.1 percentage units greater in NDF and 3.45 percentage units less in starch content than the HI treatments. Intake of DM was greater for heifers fed 3335-HI than 3335-LO; however, DMI was greater by heifers fed 3223-LO than 3223-HI (hybrid x cutting height interaction, P < 0.05). Starch intake was greater (P < 0.05) and NDF intake was less (P < 0.05) by heifers fed HI vs. LO and fed 3335 vs. 3223 dietary treatments. Digestibility of DM, starch, and NDF was greater (P < 0.05) by heifers fed 3223 than 3335 dietary treatments, but digestibility differences were not observed (P > 0.10) between cutting heights. Rate of in situ DM and starch degradability was not affected (P > 0.10) by hybrid or cutting height; however DM degradability was greater (P < 0.05) for HI than LO corn silage substrates at 8, 16, and 24 h of incubation. Rate of NDF degradability tended (P = 0.08) to be greater for 3223 than for 3335, and for LO compared with HI corn silage. Degradability of NDF was greater (P < 0.05) for 3223 compared with 3335 substrates at 24, 36, and 48 h of incubation. These data suggest fiber may not be an accurate measure of corn silage quality. Whereas cutting height affected chemical composition, we observed genetics to have a greater effect on corn silage quality.     

Supplementation to meet metabolizable protein requirements of primiparous beef heifers: I. Performance,forage intake,and nutrient balance

Three experiments were conducted to evaluate the response of supplementing primiparous heifers based on the metabolizable protein (MP) system during pregnancy and lactation. In Exp. 1, 12 pregnant, March-calving heifers (432 +/- 10 kg) grazing Sandhills range were randomly allotted to one of two treatments: supplementation based on either the MP system (MPR) or the CP system (CPR). Supplements were fed to individual heifers from October to February and no hay was offered. Grazed forage organic matter intake (FOMI) was measured in November, January, and February. In Exp. 2, 18 heifers (424 +/- 8 kg) were randomly allotted to one of three treatments: 1) supplementation based on the MP system with hay fed in January and February (average 2.0 kg/d; MPR/hay), 2) supplementation based on the CP system, with hay fed in January and February (CPR/hay), or 3) supplementation based on the MP system, with no hay fed (MPR/no hay). Supplements were fed from October to February, and FOMI was measured in December and February. In Exp. 3, lactating 2-yr-old cows (394 +/- 7 kg) maintained on meadow hay were supplemented to meet either 1) MP requirements (LMPR) or 2) degradable intake protein requirements (LDIPR). Body weight (BW) and body condition score change, hay intake, and milk production were measured. In Exp. 1, grazed FOMI decreased (P = 0.0001) from 1.9% of BW in November to 1.2% in February, but no differences among treatments were detected for FOMI or BW change. In Exp. 2, grazed FOMI declined (P = 0.0001) from 1.7% of BW in December to 1.1% in February, with no differences among treatments. Heifers on the MPR/hay and CPR/hay treatments had higher (P = 0.0018) total intake (grazed forage + hay intake) in February (1.7% BW) than the MPR/no hay heifers (1.1% BW). Heifers on the MPR/no hay treatment had a lower weight (P = 0.02) and tended (P = 0.11) to have a lower BCS than heifers on other treatments. In Exp. 3, the LMPR cows had higher (P = 0.02) ADG than LDIPR cows (0.41 and 0.14, respectively), but treatment did not affect milk production. Organic matter hay intake averaged 2.4% of BW. We conclude that supplementation to meet MP requirements had little benefit to heifer performance during gestation, but increased weight change during lactation. Because grazed forage intake decreased from 1.9 to 1.1% of BW with advancing gestation, supplemental energy is necessary to reduce weight and condition loss of gestating hefiers grazing dormant Sandhills range.     

Effects of locoweed on serum swainsonine and selected serum constituents in sheep during acute and subacute oral/intraruminal exposure

A study was conducted to evaluate the effects of acute and subacute locoweed exposure on serum swainsonine concentrations and selected serum constituents in sheep. Thirteen mixed-breed wethers (BW = 47.5 +/- 9.3 kg) were assigned randomly to 0.2, 0.4, or 0.8 mg of swainsonine x kg BW(-1) x d(-1) treatments. During acute (24 h) and subacute (19 d) exposure, serum swainsonine was detected in all treatments and was greatest (P < 0.03) in the 0.8 mg treatment. Serum alkaline phosphate (ALK-P) activity was increased (P < 0.01) for the 0.8 mg treatment compared with baseline (0 h) by 7 h and continued to increase throughout the initial 22 h following acute exposure to locoweed. A linear increase (P < 0.01) in serum ALK-P activity was noted, with the rate being 3.00 +/- 0.56 U x L(-1) x h(-1). Serum ALK-P activity was increased (P < 0.05) across treatments on d 7 over d -19, -12, 0, 1, 21, and 26; on d 14 over d -19, -12, 0, and 26; and on d 19 over d -19, -12, 0, 1, 21, and 26. By d 20, approximately 48 h after last exposure to swainsonine, serum ALK-P activities were no longer different (P = 0.13) than baseline (d -19, -12, and 0), and by d 26 values had generally returned to baseline. No linear (P = 0.98), quadratic (P = 0.63), or cubic effects of swainsonine with time from exposure were noted for serum aspartate aminotransferase. Similar to serum ALK-P activities, serum aspartate aminotransferase activities were increased (P < 0.05) across treatment levels on d 7, 14, 19, 20, 21, and 26 over those on d -19, -12, 0, and 1. Total serum Fe was decreased (P < 0.05) within the initial 22 h following the swainsonine exposure. On d 21 (48 h after swainsonine feeding ended), serum Fe increased to 472 mg/L. Concentrations of ceruloplasmin were lower (P < 0.10) on d 14 and 19 following exposure to locoweed. Recovery of ceruloplasmin levels coincided with similar changes in serum Fe. There was a linear (slope = 0.33 mg x dL(-1) x d(-1); P < 0.01) effect with time of exposure to locoweed (i.e., swainsonine) on serum triglyceride concentrations. Rapid changes in serum ALK-P and Fe concentrations without parallel changes in other damage markers indicate that acute exposure to swainsonine induces metabolic changes that may impair animal production and health before events of cytotoxicity thought to induce clinical manifestation of locoism.     

Conditioning taste aversions to locoweed (Oxytropis sericea) in horses

Locoweed (Oxytropis sericea) is a serious poisoning problem for horses grazing on infested rangelands in the western United States. Our objectives were to determine 1) whether lithium chloride or apomorphine would condition aversions to palatable foods, and at what doses, and 2) whether horses could be averted to fresh locoweed in a pen and grazing situation. Apomorphine was not an acceptable aversive agent because at the dose required to condition an aversion (> or = 0.17 mg/kg BW), apomorphine induced unacceptable behavioral effects. Lithium chloride given via stomach tube at 190 mg/kg BW conditioned strong and persistent aversions to palatable feeds with minor signs of distress. Pen and grazing tests were conducted in Colorado to determine if horses could be averted to fresh locoweed. Pen tests indicated that most horses (5/6) were completely averted from locoweed. Treated horses ate 34 g of fresh locoweed compared to 135 g for controls (P < 0.01) during three pen tests when offered 150 g per test. One horse (T) in the treatment group ate locoweed each time it was offered in the pen, but ate no locoweed while grazing. In the grazing trial, control horses averaged 8.6% of bites of locoweed (P < 0.01) during the grazing portion of the study, whereas treated horses averaged <0.5%. One treated horse (S) accounted for all consumption; he consumed 15% of his bites as locoweed in a grazing bout on d 2 of the field study. Thereafter, he was dosed a second time with lithium chloride and ate no locoweed in the subsequent 5 d. Three of six horses required two pairings of lithium chloride with fresh locoweed to condition a complete aversion. The results of this study indicate that horses can be averted from locoweed using lithium chloride as an aversive agent, and this may provide a management tool to reduce the risk of intoxication for horses grazing locoweed-infested rangeland.     

Nitrogen metabolism of beef steers fed endophyte-free tall fescue hay: effects of ruminally protected methionine supplementation

Level of nitrogen (N) intake and ruminally protected methionine supplementation were evaluated in eight Angus growing steers (initial BW 253+/-21 kg, final BW 296+/-21 kg) in a replicated 4+/-4 Latin square design. The steers were fed two endophyte-free tall fescue (Festuca arundinacea) hays that contained 2.2 (LO) or 2.8% (HI) of DM as N and were either supplemented or not with ruminally protected methionine (10 g metabolizable methionine/d). Diets were fed to provide adequate energy for 0.5 kg ADG and sufficient protein for maintenance (LO), or protein to support 0.5 kg ADG (HI). Following at least 14 d of adjustment, N balance was measured for 6 d. Isotopic urea was infused (15N15N-urea, 0.164 mmol urea N/h) via a jugular catheter for 56 h and urine was collected from 48 to 56 h to measure urea kinetics. Jugular blood was collected during the balance trial, and serum was analyzed for serum urea N (SUN). By design, daily N intake was greater (P < 0.05) for HI (112 g) than for LO (89 g). Compared with LO, steers when fed HI had greater (P < 0.05) daily DMI (4,217 vs 4,151 g), fecal N (34.4 vs 31.2 g), N digested (77.1 vs 57.7 g), urine N (48.3 vs 37.5 g), urine urea N excretion (34.6 vs 24.8 g), and N retained (29.8 vs 21.1 g). When fed HI steers also had higher (P < 0.05) urine urea N concentration (276 vs 219 mM), SUN (8.7 vs 6.7 mM), N digestibility (69.1 vs 64.9%), percentage of urinary N present as urea (71.5 vs 66.7%, P < 0.053), and rate of urea N production (59.6 vs 49.2 g/d) but lower (P < 0.05) percentage of urea N produced that was returned to the ornithine cycle (15.03 vs 19.2 1%) than when fed LO. Methionine supplementation decreased daily urine N (41.2 vs 44.6 g, P = 0.10) and increased both the amount of N retained daily (27.9 vs 23.7 g, P < 0.089) and the percentage of N digested that was retained (40.4 vs 34.6%, P < 0.094). In summary, supplemental methionine met a specific dietary limitation by increasing the amount of digested N that was retained by the steers.     

Protein expression in dairy cows with and without subclinical hypocalcaemia

AIM: To determine differences in plasma proteomic profiles between healthy cows and those with subclinical hypocalcaemia within 12 hours after calving, and thereby explore the underlying biological mechanism of subclinical hypocalcaemia in dairy cows.

METHODS: Plasma samples were collected within 6 hours of calving from Holstein cows on a farm in Heilongjiang, China; 32 with subclinical hypocalcaemia (plasma calcium concentration 1.38–2.00?mmol/L and no clinical signs) and 59 control cows (plasma calcium concentration 2.10–2.8?mmol/L). Plasma samples were applied to weak cationic exchange protein chips for protein profiling by surface-enhanced laser desorption/ionisation time-of-flight mass spectrometry (SELDI-TOF-MS), and the data were analysed using the PBS-IIC system. The amplitude of peaks for the two groups were compared using the Wilcoxon sum-rank test, and the mass-to-charge ratio of the peaks that differed was used to identify peptide fragments using the Swiss-Prot protein database.

RESULTS: Seven peaks were identified in the subclinical hypocalcaemia group that differed from those of the control group (p<0.001), that represented six unique proteins. Expression of serum albumin, fibrinogen alpha chain, amyloid beta A4 proteins and neurosecretory protein VGF were increased, and expression of apolipoprotein A-II and serum amyloid A proteins were decreased in the subclinical hypocalcaemic cows compared with control cows.

CONCLUSION: Use of SELDI-TOF-MS technology can effectively identify differences in plasma protein expression patterns in cows with subclinical hypocalcaemia. Neurosecretory protein VGF and amyloid beta A4 protein might represent useful biomarkers for diagnosis of subclinical hypocalcaemia.     

Targeted Grazing of White Locoweed: Short-Term Effects of Herbivory Regime on Vegetation and Sheep

White locoweed (Oxytropis sericea Nuttall) and nontarget vegetation response to 2 yr of targeted grazing by sheep, one treatment of picloram plus 2, 4-D (HER) or no treatment (CON) were compared. Serum of sheep that grazed locoweed intermittently (IGZ, 5 d on locoweed followed by 3 d off locoweed) vs. counterparts that grazed locoweed continuously for 24 d (CGZ) was also examined. Alkaloid toxicity was inferred by serum levels of thyroxine (T4), triiodothyronine (T3), alkaline phosphatase (ALKP), aspartate aminotransferase (AST), and swainsonine, as well as behavior and body weight gains. Three sites were used in a randomized complete block design. IGZ, CGZ, and HER treatments reduced locoweed density (P < 0.01), canopy cover (P < 0.01), number of flower stalks (IGZ: P = 0.02, CGZ and HER: P = 0.01), and plant size (P < 0.01). White locoweed seed density in the soil seed bank was not reduced with grazing, and nontarget vegetation was mostly unaffected by treatments. Grass canopy cover increased in grazed and herbicide plots throughout the study (IGZ: P = 0.03, CGZ and HER: P < 0.01). Percentage bare ground was unchanged (IGZ: P = 0.46, CGZ: P = 0.44) in grazed plots but decreased (P = 0.03) in HER plots. After 24 d, ewes in the IGZ treatment had lower levels of serum ALKP (P < 0.01) and AST (P = 0.02) and marginally lower swainsonine levels (P < 0.07) than CGZ ewes that tended to exhibit lower serum T3 (P < 0.07) and similar serum T4 (P = 0.25) levels. Time spent feeding on locoweed tended to differ (P = 0.06) between treatments. Body weight gain was the same (P = 0.19) regardless of treatment. IGZ of locoweed-infested rangeland with sheep may be a viable short-term means of reducing locoweed density without detrimentally affecting animal health.     

Effects of central administration of glucagon on feed intake and endocrine responses in sheep

This study was conducted to investigate effects of glucagon intracerebroventricularly administered on feed intake and endocrine changes in sheep. Four male sheep (48–55 kg BW) were used. The animals were acclimatized to be fed alfalfa hay cubes at 12.00 hour. Human glucagon (40 and 80 µg/0.5 mL) was injected into the lateral ventricle at 12.00 hour. Blood samples were taken every 10 min from 30 min before to 180 min after the glucagon injection. Soon after the injection, the animals were given alfalfa hay cubes, and the amounts of the feed eaten within 2 h were measured. Feed intakes were significantly (P < 0.05) suppressed by 80 µg of glucagon. Plasma glucose levels in control animals were gradually decreased after the feeding, whilst those in glucagon‐treated animals were temporarily elevated just after the feeding and then kept higher than control levels. Plasma insulin was abruptly elevated after the feeding and was maintained at higher levels than before the feeding in all treatments. Plasma NEFA concentrations were decreased after the feeding in all treatments. A tendency of increase in plasma cortisol levels occurred in glucagon‐injected animals. The present study provides the first evidence that glucagon directly acts on the brain, then inhibiting feeding behavior and inducing endocrine responses in ruminants.     

Urea flux in beef steers: effects of forage species and nitrogen fertilization

The effects of two forage species and N levels on urea kinetics and whole-body N metabolism were evaluated in eight Angus steers (initial BW 217+/-15 kg). In a replicated, 4 x 4 Latin square design, steers were fed gamagrass (Tripsacum dactyloides L.) or switchgrass (Panicum virgatum L.), each of which had 56.2 (LO) or 168.5 (HI) kg of N fertilization per hectare. Diets provided adequate energy for 0.5 kg ADG. Nitrogen balance and urea kinetics were measured from d 22 to 27 of each period. Urine samples collected during intravenous infusion of bis 15N urea were used to calculate production and recycling of urea N from relative abundance of urea isotopomers. Jugular blood serum was analyzed for serum urea N (SUN). Gamagrass differed from switchgrass (P < 0.05) in daily DMI (4,273 vs 4,185 g), N intake (72 vs 67 g), DM digestibility (61.0 vs 63.6%), fecal N (30.6 vs 28.3 g/d), urine urea N (10.5 vs 8.0 g/d), and percentage of urinary N present as urea N (53.5 vs 40.0%). After adjustment for differences in N intake, fecal N still tended to be greater (P < 0.09) for gamagrass than for switchgrass. The LO differed from the HI (P < 0.01) in daily N intake (63 vs 76 g), DM digestibility (61.3 vs 63.3%), urine N (13.6 vs 25.9 g/d), and N retained as a percentage of N digested (57.3 vs 43.5%). Compared to switchgrass, gamagrass had greater SUN, N digestibility, and N digested as N level increased (forage x N level interactions, P < 0.05). As N level increased, N retention increased from 19.5 to 23.5 g/d in gamagrass and decreased from 20.5 to 18.1 g/d in switchgrass (interaction, P < 0.07). The HI group was greater than the LO intake group (P < 0.03) in endogenous production of urea N (44.4 vs 34.0 g/d), gut entry rate of urea N (31.6 vs 28.2 g/d), and the amount of urea N that re-entered the ornithine cycle (9.4 vs 7.9 g/d). However, the percentage of urea N entering the gastrointestinal tract that was recycled was constant among treatments (29.1%), indicating that almost 70% of the urea N that entered the gastrointestinal tract was potentially available for anabolic purposes of the steers as a component of microbial products that were absorbed or excreted in the feces. In summary, N levels affected N metabolism of steers more when they were fed gamagrass than when they were fed switchgrass. Although the absolute amounts of N moving through the system changed with variations in intake, the proportions remained similar, with a greater efficiency of N use at low N intakes.     

Substitution of fresh forage ramie for alfalfa hay in diets affects production performance,milk composition,and serum parameters of dairy cows

We hypothesized that ramie, Boehmeria nivea (a nettle native to Asia), can be used as a high-quality forage to replace alfalfa hay in diets of dairy cows. Accordingly, we aimed to examine the effects of substituting fresh forage ramie for alfalfa (Medicago sativa) hay on production performance, milk composition, and serum parameters of dairy cows. Thirty-two Holstein cows (body weight (BW)?=?590?±?50.6 kg) were randomly divided into four groups of eight cows. The experimental period lasted 10 weeks. The dietary treatments consisted of four proportions of fresh forage ramie (0, 33, 67, and 100%) as a substitute for alfalfa hay (designated as CON, FR33, FR67, and FR100, respectively). On days 69 and 70 of the experimental period, milk and blood samples were collected for analysis. We found no significant differences in the milk yield and milk quality (milk protein percentage, milk fat percentage, and milk lactose percentage) between the treatments. We did find that forage ramie significantly reduced dry matter intake (DMI) and 4% fat-corrected milk (FCM) yield. Meanwhile, no significant differences were detected in serum parameters between the treatments, with the exception of triglyceride (TG) and alanine aminotransferase (ALT). In conclusion, our data indicate that alfalfa hay can be replaced with forage ramie in the diet of Holstein cows in the milk lactation stage, with no negative effects on milk quality and blood parameters.

     

Milk yield, milk composition, eating behavior, and lamb performance of ewes fed diets containing soybean hulls replacing coastcross (Cynodon species) hay

The objective of this experiment was to evaluate the effects of replacing coastcross hay NDF by soybean hull (SH) NDF on the lactation performance and eating behavior of ewes and also on the performance of their lambs. Fifty-six Santa Inês lactating ewes (56.1 +/- 6.8 kg of initial BW; mean +/- SD) were penned individually and used in a randomized complete block design with 14 blocks and 4 treatments. Diets were formulated to provide similar concentrations of NDF (56%) and CP (16%). The SH NDF replaced 33 (SH33), 67 (SH67), or 100% (SH100) of the NDF contributed by coastcross hay in a 70% forage-based diet (SH0), resulting in SH inclusion rates of 0, 25, 54, and 85% of the dietary DM. Once a week, from the second to the eighth week of lactation (weaning time), ewes were separated from their lambs, stimulated by a 6-IU i.v. oxytocin injection, and hand milked to empty the udder. After 3 h, milk production was obtained after the same procedure. Quadratic effect for milk production (142.4, 179.8, 212.6, and 202.9 g/3 h) and cubic effect for DMI (2.27, 2.69, 3.25, and 3.00 kg/d) were observed as SH inclusion increased from 0 to 85% of the dietary DM. Milk fat (7.59, 7.86, 7.59, and 7.74%), protein (4.53, 4.43, 4.40, and 4.55%), and total solids (18.24, 18.54, 18.39, and 18.64%) did not differ among the 70% forage-based diet and diets with SH NDF replacing 33, 67, or 100% of the NDF. A linear increase in lactose concentration was observed with SH inclusion. Ewe BW gain during the trial showed a cubic response (0.37, 0.03, 4.80, and 2.80 kg) with SH inclusion. The preweaning ADG of lambs increased linearly, and ADG of lambs after weaning decreased linearly with SH inclusion. Final BW of lambs (2 wk after weaning) did not differ among treatments. Eating behavior observations were conducted with 44 ewes. The same facilities, experimental design, dietary treatments, and feeding management were used. Observations were visually recorded every 5 min for a 24-h period when ewes were 46 +/- 6.8 d in milk. Eating time (min/d, min/g of DMI, and min/g of NDF intake) and time expended in rumination and chewing activities (min/g of DMI and min/g of NDF intake) decreased linearly with the addition of SH in the diets. The inclusion of SH improved DMI and milk production, also reflecting on the BW of lambs at weaning. Milk performance was not affected when SH NDF replaced 100% of hay NDF.     

Nutritional and toxicological evaluations of kochia hay (Kochia scoparia) fed to lambs

Kochia foliage that had tested positive to Dragendorff's reagent (presumptive alkaloids) and had elicited chronic toxicosis when fed to rats was fed to sheep to characterize early stages of kochia toxicosis and evaluate treatments that might improve tolerance. Twelve fine-wool lambs (46 +/- 9 kg BW) were fed chopped kochia hay (35%) mixed with chopped alfalfa hay (65%) for 4 wk. The kochia diet had 14.3% CP and 39.9% ADF. Dry matter intake averaged 3.4% of BW/d. Body weight did not change during 4 wk and blood serum components were not changed from values at the onset. Thereafter, kochia was increased to 50% of diet for five more weeks, during which four treatments were imposed randomly (three lambs/treatment): 1) none; 2) N-acetyl-L-cysteine plus trans-stilbene oxide, 21 and 52 mg/kg of BW, respectively, given i.p. twice weekly; 3) retinyl palmitate, 275 mg, plus alpha-tocopherol, 300 mg/lamb dosed i.m. twice weekly; and 4) zinc sulfate mixed in the feed to provide 500 mg daily. Kochia contained 4.8% oxalate. The diet with 50% kochia had 16% CP and 36% ADF, and digestibility coefficients were 59% for DM, 72% for CP, and 59% for ADF. After 5 wk, blood glucose was elevated slightly, total bilirubin was increased about 1.5-fold (P less than .05), alanine aminotransferase was elevated slightly (P less than .05), and inorganic phosphorus and urea (blood urea N) were diminished (P less than .05); other serum components, including calcium, were unchanged from initial levels (P greater than .10). Treatments had negligible effects for modifying serum signs of mild chronic toxicosis associated with kochia hay fed as 50% of diet.     

Altered metabolic hormones, impaired nitrogen retention, and hepatotoxicosis in lambs fed Kochia scoparia hay

Livestock grazing lush Kochia scoparia (L.) Schrad, sometimes experience BW loss, hyperbilirubinemia, photosensitization, and polyuria. Animals fed kochia hay may exhibit milder or negligible signs of toxicosis but fail to utilize nutrients efficiently. To characterize early aspects of kochia toxicosis and to evaluate prospective treatments, 12 wether lambs (34 +/- 3 kg) were fed prebloom kochia hay (83% OM, 15% CP, and 6.3% total oxalate) and treated as follows: 1) no treatment; 2) drenched daily with aqueous ZnSO4 to provide 30 mg of Zn/kg of BW); 3) injected i.p. twice weekly with N-acetyl-L-cysteine (CYS) in saline (21 mg/kg of BW) plus trans-stilbene oxide (TSO) in corn oil (27 mg/kg of BW); and 4) treated as 2) plus 3). Treatments were imposed factorially (2 x 2) with three lambs per treatment. Kochia intake (ad libitum) averaged .57 kg/d (1.7% of BW) for 80 d, and digestibility of DM and CP were 44 and 59%, respectively, at wk 4, but BW loss was severe (6 to 11 kg/lamb). After 14 d, serum insulin and prolactin were decreased (P less than .05) below initial values (.48 to .11 and 102 to 28 ng/ml, respectively). Serum somatotropin increased (P less than .05) from 4.5 to 6.8 ng/ml at 4 wk. Serum total bilirubin increased threefold at 3 wk (P less than .05) and declined slightly thereafter through 10 wk. Early changes in serum enzymes reflected mild hepatotoxicosis without cholestasis, whereas histopathology (at 80 d) showed diffuse hepatocyte swelling and nephrosis.(ABSTRACT TRUNCATED AT 250 WORDS)