An evaluation of ruminally degradable intake protein and metabolizable amino acid requirements of feedlot calves

Ruminally degradable intake protein (DIP) and metabolizable indispensable amino acid (MIAA) requirements of feedlot steers were evaluated. Dietary treatments consisted of isocaloric 80% concentrate steam-flaked corn-based diets containing either .8% urea, 1.5% fish meal (FM), 3.0% FM, 4.5% FM, or 4.5% soybean meal (SBM). Treatment effects on characteristics of ruminal and total tract digestion were evaluated using four Holstein steers (249 kg) with cannulas in the rumen and proximal duodenum. Ruminal digestibility of OM (RDOM; P < .05) and feed N (P < .01) and microbial N flow (MNF; P < .01) to the small intestine were greater with urea as the supplemental N source. The level of DIP was closely associated (R2 = .89) with MNF. Postruminal digestibility of OM was greater (P < .05) for FM than for urea-supplemented diets, compensating for lower RDOM. There were no treatment effects (P > .10) on DOM. As the level of FM was increased, MIAA increased linearly (P < .01). Intestinal MIAA were similar (P > .10) for urea- and SBM-supplemented diets. Treatment effects on 56-d growth performance were evaluated using 100 medium-framed crossbred steers (231 kg). Daily weight gain (linear effect; P < .01), DM intake (linear effect; P < .10), feed efficiency (linear effect; P < .05), and diet NE (linear effect; P < .05) increased with level of FM supplementation. Daily weight gain (P < .10) and DM intake (P < .05) were greater for urea- than for SBM-supplemented diets. Using bovine tissue as the reference protein, the biological value (based on chemical score) of the intestinal chyme protein averaged 73%; methionine was first-limiting. There was a close association (R2 = .99) between methionine supply to the small intestine and observed/expected dietary NE. The metabolizable methionine requirement (MMETR, g/d) of medium-framed feedlot steers can be reliably predicted from measures of BW and ADG (MMETR = 1.565 + .0234ADG[268 - (29.4 x .0557BW(.75)ADG(1.097))/ADG] + .0896BW(.75)). There was a very close association (R2 = .89) between DIP and MNF (MNF = 13.7DIP - .66DIP(2) + 25.9). At maximal observed synthesis, DIP accounted for 76% of the MNF. A minimum of 100 g DIP/kg of total tract digestible OM was required to maximize RDOM and MNF.     

Body composition and metabolic profiles associated with puberty in beef heifers

Rapid growth large frame (RL, n = 61) or average growth medium frame (AM, n = 71) biotype heifers fed to achieve either moderate (MOD, .6 kg/d) or high ADG (HI, 1.0 kg/d) were used to determine whether puberty occurs at similar body composition or metabolic status. A heifer was considered pubertal after being detected in estrus and then forming a functional corpus luteum. Live animal estimates of body composition and blood samples for assessment of metabolic status were taken at 13 +/- .2 d after estrus for all heifers. Body composition and metabolic status were assessed every 56 d from 7 mo of age until puberty in a subset of 80 heifers representing all biotype-diet combinations. At puberty, 32 of these 80 heifers were slaughtered and physical and chemical composition of the empty body were determined. High-gain diet heifers were younger, heavier, taller, and more muscular (all P < .01) at puberty than MOD heifers. Slaughter measurements paralleled live animal estimates; bodies of HI and RL heifers contained more (P < .01) carcass and noncarcass components than those of MOD and AM heifers, respectively. Carcasses of RL and HI heifers were more (P < .05) muscular and fatter than AM and MOD heifers. At puberty, HI heifers had a greater (P < .01) mass of moisture, fat, and fat-free organic matter (FFOM) than MOD, whereas RL heifers had more moisture, ash, and FFOM than AM. Percentage of fat was greater (22.1 +/- 1.0 vs 1.0 vs 19.1 +/- 1.0; P < .05) and percentage of moisture was less (55.4 +/- .6 vs 58.1 +/- .6; P < .01) in bodies of HI than in those of MOD heifers. Concentrations of blood urea nitrogen and insulin were greater (P < .05) in HI than in MOD heifers. Diet did not influence concentration of IGF-I or glucose, and metabolic markers were unaffected by biotype. No dramatic changes in body composition or metabolic signals were detected before puberty. Puberty did not occur at similar body composition or metabolic status in all heifers.     

Effect of long-term cocaine administration to pregnant ewes on fetal hemodynamics, oxygenation, and growth

OBJECTIVE: To assess uterine and fetal blood flows by Doppler velocimetry and fetal growth and oxygenation in pregnant ewes treated daily with cocaine and to determine whether cocaine impairs fetal cardiac and cerebral reactivity. METHODS: The study groups received 70 mg (n = 7) or 140 mg (n = 7) of cocaine and the control group (n = 7) received placebo injected intramuscularly daily on days 60-134. Hemodynamic data were measured at rest and during two acute hypoxic tests at cesarean delivery performed on day 134. RESULTS: The fetal heart rate (FHR) and umbilical and uterine resistance indices (RIs) were higher in the cocaine groups than in the control group (FHR: 187 +/- 8 and 166 +/- 8 beats per minute at 83 and 123 days, respectively, in controls and 9-11% higher in cocaine groups; umbilical RI: 0.79 +/- 0.06, 0.60 +/- 0.04, and 0.52 +/- 0.06, at 83, 105, and 123 days, respectively, in controls and 11-17% higher in the cocaine groups [P < .01]; and uterine RI: 0.40 +/- 0.05, 0.40 +/- 0.04, and 0.37 +/- 0.04, at 83, 105, and 123 days, respectively, in controls and 13-35% higher in cocaine groups [P < .05]). At delivery on day 134, the following characteristics were found to be different in the cocaine groups: fetal weight (4.03 +/- 0.2 kg in controls and 15-21% lower in the cocaine groups [P < .02]), partial pressure of oxygen (26.5 +/- 1.4 mmHg in controls and 15-16% lower in cocaine groups [P < .05]), umbilical RI (0.40 +/- 0.03 in controls and 11-17% higher in cocaine groups [P < .01]), cerebral RI (0.61 +/- 0.03 in controls and 9-15% lower in cocaine groups [P < .01]), and cerebral-umbilical ratio (1.52 +/- 0.04 in controls and 22-23% lower in cocaine groups [P < .001]). During the hypoxic tests, the cerebral RI (P < .05) and the cerebral-umbilical ratio (P < .05) decreased significantly less in the two cocaine groups. The FHR response was reduced significantly in the two cocaine groups (P < .05). CONCLUSION: Long-term exposure to cocaine induces uterine and fetal blood flow disorders, fetal growth restriction, and hypoxia. It reduces the capability of the cerebral vessels to vasodilate and the heart rate to increase during acute hypoxia.     

The feeding value of dry-rolled and steam-flaked corn in finishing diets for feedlot cattle: influence of protein supplementation

We used 80 medium-framed yearling crossbred heifers (357 kg) in a 110-d trial to evaluate the influence of dietary protein level (11 vs 14%) on the feeding value of dry-rolled (DRC) and steam-flaked corn (SFC). All diets contained 1% urea; cottonseed meal (CSM) was the source of supplemental undegradable intake protein (UIP). Steam flaking corn reduced DMI (9%, P < .10) and increased (P < .01) feed efficiency (14%), dietary NEm (13%), and NEg (15%). Steam flaking increased the NEm by 17% and NEg by 19%. Supplemental CSM decreased (P < .10) feed efficiency (7%) and dietary NEm (4%) and NEg (6%). There were no treatment effects (P > .10) on carcass characteristics. Steam flaking corn increased (P < .05) fecal pH and reduced (P < .01) fecal starch. Supplemental CSM increased (P < .01) fecal pH and reduced (P < .01) fecal starch. Four Holstein steers (413 kg) were used in a 4 x 4 Latin square experiment to evaluate treatment effects on digestive function. Steam flaking corn increased (P < .05) flow of nonammonia N (11%, P < .05) and microbial N (15%, P < .01) to the duodenum. Supplemental CSM increased the flow of microbial N (6%, P < .01), feed N (21%, P < .10), and nonammonia N (12%, P < .05) to the duodenum. The UIP value of CSM was 28% for the DRC diet and 52% for the SFC diet. Steam flaking corn increased (P < .01) ruminal starch digestion (26%) and total tract digestibility of OM (17%), N (15%), starch (19%), and GE (17%). Steam flaking increased the DE value of corn by 21%. Supplemental CSM did not influence (P > .10) postruminal or total tract starch digestion. Supplemental CSM decreased (7%, P < .10) the DE value of the diet. We conclude that increasing the postruminal protein supply of a corn-based finishing diet beyond that provided by urea supplementation, alone, will not enhance starch digestion or the energy value of the diet.     

Review: effect of protein nutrition on ovarian and uterine physiology in dairy cattle

Milk production and dry matter intake of dairy cows are stimulated in response to increased intake of dietary protein, but, unfortunately, decreased fertility is often associated with this nutritional strategy. Ruminally degradable protein or ruminally undegradable protein in excess of requirement can contribute to reduced fertility in lactating cows. Dietary protein nutrition or utilization and the associated effects on ovarian or uterine physiology have been monitored with urea nitrogen in plasma or milk; concentrations above 19 mg/dl have been associated with altered uterine pH and reduced fertility in dairy cows. The uterine pH changed dynamically and inversely with plasma urea nitrogen, signaling possible changes in the uterine milieu. Mechanisms for reduced fertility include exacerbation of negative energy balance and reduced plasma progesterone concentrations when cows were fed rations that were high in ruminally degradable intake protein. Alternatively, changes in uterine secretions that are associated with high protein intake and elevated plasma urea nitrogen might be detrimental to embryos. Bovine endometrial cells in culture respond directly to increasing urea concentrations with alteration in pH gradient but respond most notably with increased secretion of prostaglandin F2 alpha (PGF2 alpha). Increased uterine luminal PGF2 alpha interferes with embryo development and survival in cows, thus providing a plausible link between elevated plasma urea nitrogen concentrations and decreased fertility. Poor fertility in high producing dairy cows reflects the combined effects of a uterine environment that is dependent on progesterone and rendered suboptimum by the antecedent effects of negative energy balance or postpartum health problems and that is further compromised by the effects of urea resulting from intake of high dietary protein.     

Effects of a dietary mixture of meat and bone meal, feather meal, blood meal, and fish meal on nitrogen utilization in finishing Holstein steers

Our objective was to determine to what extent rate and efficiency of protein gain in finishing cattle can be enhanced by feeding an amino acid-balanced mixture of undegraded intake proteins. The Cornell Net Carbohydrate and Protein System (CNCPS) model was used to formulate a corn-based diet that would meet the rumen requirements for 410-kg large-framed steers with an estrogen implant and fed an ionophore. The CNCPS model was also used to formulate a highly undegradable intake protein (UIP) mixture from meat and bone meal, blood meal, fish meal, and hydrolyzed feather meal to provide the amino acids needed to supplement those derived from microbial protein to better meet amino acid requirements for growth. Four Holstein steers weighing 407 kg were offered a 90:10 concentrate-forage diet at hourly intervals at 95% of ad libitum intake. The steers were injected with 500 microg of estradiol-17beta at 12-h intervals to mimic the effects of an estrogenic implant. Treatments planned consisted of inclusion of the UIP mixture at 0, 2.5, 5, and 7.5% of the diet DM. Dry matter intake was fixed at 6.4 kg/d, and DM digestibility was not significantly affected by varying the amount of UIP addition. Apparent digestibility of N increased (P = .011) from 63.8 to 65.8, 70.7, and 71.5%, the amount of N absorbed increased (P = .001) from 73 to 84, 100, and 106 g/d, and N balance increased (P = .003) from 20 to 30, 33, and 39 g/d when UIP was fed at 0, 2.6, 5.2, and 7.8% of diet DM, respectively. The efficiency of N use increased 39.7%, and biological value increased 31.6% when the UIP mixture was added to the diet. Circulating concentrations of plasma urea N (PUN) were increased (P = .017) from 4.5 for the control diet to 5.7, 6.2, and 6.1 mg/dL when the UIP mixture was added at 2.6, 5.2, and 7.8%, respectively. Corresponding IGF-I concentrations were also increased from 491 to 558 and 624 ng/mL with 2.6 and 5.2% levels of UIP addition. Plasma glucose, NEFA, and insulin concentrations were not affected by feeding the UIP mix. The rate and efficiency of N use for growth improved with addition of an amino acid-balanced UIP mixture to the diet.     

Development of a progestin-based estrus synchronization program: II. Reproductive response of cows fed melengestrol acetate for 14 days with injections of progesterone and prostaglandin F2alpha

We tested the efficacy of an estrus control system designed to provide optimal control of follicular development. In Exp. 1, postpartum cows (n = 133) and yearling heifers (n = 57) were fed either .5 mg x female(-1) x d(-1) of melengestrol acetate (MGA) or the carrier for MGA from d -13 to d 0 (d 0 = last day of MGA feeding). All females received 25 mg of PGF2alpha (i.m.) on d -13 and 0. On d -6, cows and heifers fed MGA were administered an i.m. injection of progesterone (200 mg; MGA/P4), and those fed the corn carrier (2XPGF2alpha) received no progesterone. Beginning on d 1, females were bred by AI from d 1 to at least d 5. During the estrus synchronization period (d 1 to d 5), more (P < .05) postpartum cows were observed in estrus (70.1 vs 42.4%), the timing of estrus was more (P < .05) precise, conception rate was similar, and pregnancy rate was higher (P < .05) in the MGA/P4 than in the 2XPGF2alpha treatment. More (P < .05) cows that were anestrous at the beginning of the breeding season were in estrus during the synchronization period in the MGA/P4 (55.8%) than in the 2XPGF2alpha (28.6%) treatment. In heifers, estrus was synchronized in over 90% of females, and neither conception nor pregnancy rate during the synchronization period differed between treatments. In Exp. 2, postpartum cows (n = 122) and heifers (n = 84) received treatments (MGA/P4 or 2XPGF2alpha) as described for Exp. 1 with one exception. In the MGA/ P4 treatment, progesterone was administered on d -7 rather than d -6. Females were bred by AI from d 1 to 5. The estrus response and conception rate during the synchronization period did not differ between treatments for either cows or heifers. We conclude that the progestin-based estrous synchronization system used in this study effectively synchronized an estrus of normal fertility in cyclic cows and induced a majority of anestrous cows to reinitiate estrous cycles.     

Concentration of tissue inhibitor of metalloproteinases (TIMP)-1 in ovine follicular fluid and serum

Tissue inhibitor of metalloproteinases (TIMP)-1 mRNA and protein localize within granulosal cells of post-gonadotropin-surge follicles and luteal tissue in ewes. Our objectives were to test the hypotheses that 1) follicular fluid concentration of TIMP-1 increases following a gonadotropin surge induced by LHRH agonist (Exp. 1) and 2) luteal status affects peripheral serum concentration of TIMP-1 (Exp. 2 and 3). In Exp. 1, the concentration of TIMP-1 within antral fluid from post-surge follicles (28.7 +/- 6.65 microg/mL) was greater (P < .02) than from pre-surge follicles (2.37 +/- 2.47 microg/mL). In Exp. 2, serum concentration of TIMP-1 did not differ among d 0 to 6 (1.27 +/- .55 microg/mL) of the estrous cycle or among periods of luteal maintenance (1.29 +/- .06 microg/mL), spontaneous luteal regression (1.19 +/- .09 microg/mL), or luteal development (1.22 +/- .08 microg/mL). However, serum concentration of TIMP-1 was greater ( P < .001) during the period of luteal maintenance (1.14 +/- .04 microg/mL) than during PGF2alpha-induced luteolysis (d 26; .85 +/- .06 microg/mL) and induced luteal absence (d 27 to 33; .95 +/- .05 microg/mL). In Exp. 3, ewes (n = 14) were bled daily from d 1 to 19 (d 0 = estrus) and at 12-min intervals for 6 h on d 3, 10, and 17. Although concentration of TIMP-1 varied considerably within and among ewes, mean concentration of TIMP-1 per ewe per day increased ( P < .05) from d 3 to 17. These data indicate that follicular fluid concentration of TIMP-1 increases following a gonadotropin surge, but the contribution of ovarian derived TIMP-1 to peripheral serum concentration is negligible.     

Development of a progestin-based estrus synchronization program: I. Reproductive response of cows fed melengestrol acetate for 20 days with an injection of progesterone

We designed two experiments to determine the efficacy of an estrus control system in cows that combined long-term progestin exposure (20 d) with an acute increase in progesterone concentration. In Exp. 1, cows (n = 30) were fed either melengestrol acetate (MGA; .5 mg x cow(-1) x d(-1)) or ground ear corn (MGA carrier) for 20 d. On d -15 (last day of MGA feeding = d 0), cows were administered 25 mg of PGF2alpha to regress the corpus luteum (CL) and establish an environment conducive to the development of persistent follicles. To synchronously regress persistent follicles, cows fed MGA (n = 15) were injected with 200 mg of progesterone on d -2 (MGA-P), and the cows fed the MGA carrier were not treated (CONT; n = 15). Cows in the CONT group were artificially inseminated 12 h after detection of spontaneous estrus from d -20 to d 8. Estrus was observed, and all cows in the MGA-P group were artificially inseminated during the period of estrus synchronization (SYNC; d 1 to 8). No difference in conception rate was observed between treatments. In Exp. 2, postpartum cows (n = 113) received either the MGA-P (n = 56) or CONT (n = 57) treatment. More (P < .05) cows were observed in estrus during SYNC in the MGA-P (50%) than in the CONT (28%) group. Of the cows in the MGA-P group that were not observed in estrus during SYNC, 50% were in estrus for the first time 23 to 29 d after MGA withdrawal (SYNC2), suggesting that these cows ovulated without observable estrus during SYNC. Estrus was observed for the first time during SYNC2 in more (P < .05) cows in the MGA-P (25%) than in the CONT (7%) group. Conception rate at the synchronized estrus, pregnancy rate, and interval to first service and pregnancy were similar between treatments. We conclude that administration of MGA-P results in the synchronization and(or) induction of a fertile estrus in cows.     

Evaluation of potato proteins on the growth performance of early-weaned pigs

We conducted five experiments to evaluate conventional and low-glycoalkaloid potato protein (CPP and LGPP, respectively) in diets for early-weaned pigs. In Exp. 1, 150 weanling pigs (initially 4.4 +/- .9 kg and 15.5 +/- 2 d of age) were fed either a control diet containing 3% spray-dried animal plasma (SDAP) or diets with additional SDAP (2.5 or 5% added; 5.5 or 8% total) or CPP (2.6% or 5.1%) substituted on a total lysine basis. From d 0 to 14 after weaning, increasing SDAP increased (linear, P < .05) ADG and ADFI, whereas increasing CCP had no effect on growth performance. In Exp. 2, 180 weanling pigs (initially 5.9 +/- 1.2 kg and 20 +/- 2 d of age) were fed diets containing a LGPP replacing 25, 50, 75, or 100% of the 7% dietary SDAP on a digestible lysine basis. From d 0 to 7 after weaning, increasing LGPP increased and then returned to control levels ADG and ADFI (quadratic, P < .01) and gain:feed ratio (quadratic, P < .05). In Exp. 3, 175 weanling pigs (initially 5.5 +/- 1.1 kg and 20 +/- 3 d of age) were fed either a control diet containing 20% dried whey, 17.5% dried skim milk, and 4% select menhaden fish meal (SMFM) or diets consisting of lactose and either 3.5 and 7.0% SDAP or 4.0 and 8.0% LGPP added at the expense of dried skim milk on a digestible lysine basis. From d 0 to 7 after weaning, ADG and ADFI increased (linear, P < .05) with increasing SDAP. With increasing LGPP, ADG and ADFI increased and then decreased (quadratic, P < .10 and P < .05, respectively). Gain:feed ratio (G/F) was not affected by SDAP and was improved (linear, P < .05) for pigs fed increasing LGPP. In Exp. 4, 270 weanling pigs (initially 6.2 +/- 1.6 kg and 20 +/- 3 d of age) were used to compare three diets that contained either 2.5% spray-dried blood meal (SDBM), 4.8% SMFM, or 3.92% CPP; test feedstuffs were substituted on a total lysine basis and diets were fed from d 7 to 28 after weaning. Pigs fed CPP had decreased (P < .05) ADG and G/F compared with those fed the other protein sources. In Exp. 5, 255 weanling pigs (initially 5.3 +/- 1.2 kg and 17 +/- 2 d of age), were used to compare five diets that contained either 2.5% SDBM, 5.51% SMFM, 4.17% CPP, 4.17% LGPP or 8.34% LGPP; feedstuffs were substituted on a digestible lysine basis and diets were fed from d 7 to 28 after weaning. No differences (P > .10) were observed in growth performance among pigs fed any of the protein sources within the experiment. However, pigs fed the LGPP had numerically greater ADG and better G/F than those fed CPP. In conclusion, these results suggest that LGPP can be an effective replacement for a portion of the SDAP in diets for weanling pigs.     

Moderate physical activity can increase dietary protein needs

Six healthy men completed three 1-hr bouts of treadmill walk-jogging at low (L; 42 +/- 3.9% VO2max), moderate (M; 55 +/- 5.6%), and high (H; 67 +/- 4.5%) exercise intensity in order to determine whether moderate physical activity affects dietary protein needs. Both sweat rate and sweat urea N loss were greater (p < .10) with increasing exercise intensity. Seventy-two hour postexercise urine urea N excretion was elevated (p < .05) over nonexercise control (26.6 +/- 2.96 g) with both M (31.0 +/- 3.65) and H (33.6 +/- 4.39), but not L (26.3 +/- 1.86), intensities. Total 72-hr postexercise urea N excretion (urine + sweat) for the M and H exercise was greater than control by 4.6 and 7.2 g, respectively. This suggests that 1 hr of moderate exercise increases protein oxidation by about 29-45 g, representing approximately 16-25% of the current North American recommendations for daily protein intake. These data indicate that the type of exercise typically recommended for health/wellness can increase daily protein needs relative either to sedentary individuals or to those who exercise at lower intensities.     

Protein balance in the first week of life in ventilated neonates receiving parenteral nutrition

BACKGROUND: Protein intake is frequently delayed in ill neonates because of concerns about their ability to metabolize substrates. OBJECTIVE: We aimed to determine the factors affecting protein balance in ventilated, parenterally fed newborns during the first week of life. DESIGN: Leucine kinetic studies were performed in 19 neonates by using the [1-(13)C]leucine tracer technique after 24 h of a stable total parenteral nutrition (TPN) regimen. TPN intakes were prescribed by rotating attending physicians, enabling assessment of protein metabolism over a range of clinically used nutrient intakes. RESULTS: Mean (+/-SD) birth weight was 1.497 +/- 0.779 kg, gestational age at birth was 30.3 +/- 4.0 wk, and age at study was 3.9 +/- 1.4 d. Amino acid intakes (AAIs) ranged from 0.0 to 2.9 g x kg(-1) x d(-1). Based on leucine kinetic data, protein balance was calculated as the difference between protein synthesis and catabolism. By multiple regression analysis, AAI was the only predictor associated independently with protein balance (P < 0.01); energy intake, lipid intake, glucose intake, birth weight, and gestational age were not. Both leucine oxidation and nonoxidative leucine disposal rates were significantly correlated with leucine intake (P < 0.0005 and P < 0.01, respectively). Of the 12 infants with AAIs > 1 g x kg(-1) x d(-1), only 1 infant was significantly catabolic (protein balance <-1 g x kg(-1) x d(-1)). There was no evidence of protein intolerance as determined by elevated creatinine (69 +/- 31 micromol/L), plasma urea nitrogen (6.7 +/- 2.53 mmol/L), or metabolic acidosis (pH: 7.36 +/- 0.05). CONCLUSIONS: Ill neonates can achieve a positive protein balance in the first days of life without laboratory evidence of protein toxicity.     

Final height after combined growth hormone and gonadotrophin-releasing hormone analogue therapy in short healthy children entering into normally timed puberty

Four estrous beagles were inseminated with 1 x 10(8) sperm into both the right and left uterine horns, and the uterine horn and oviduct on one side were removed under anesthesia after 7 hr and 24 hr, respectively. The lumen of the uterine horns and oviducts was flushed with canine capacitation medium (CCM), and movement of the sperm in CCM was assessed by phase-contrast microscopy. In a second experiment, ejaculated sperm obtained from 5 normal beagles was incubated in CCM supplemented with oviductal flush fluid (OF-CCM) at 38 degrees C with 5% CO2 in air. Motility of sperm, and percentages of hyperactivated sperm (%HA) and acrosome-reacted sperm (%AR) among freely swimming (FS) sperm were investigated until 24 hr after the start of incubation. After 7 hr of incubation the sperm was coincubated with canine oocytes in OF-CCM for 2 hr, and the number of zona pellucida-binding (zona-binding) sperm was then counted. The %HA among the sperm in the oviductal flush fluid both 7 hr (mean +/- S.E.; 15.0 +/- 2.4%) and 24 hr (77.5 +/- 5.2%) after intrauterine insemination were significantly higher than in the uterine flush fluid (P < 0.05, 0.01, respectively). The motility and %HA among FS-sperm in OF-CCM were higher than in the control medium without oviductal fluid. However, there was no difference in the %AR between OF-CCM and control medium. The number of zona-binding sperm in OF-CCM (8 +/- 1) was significantly greater than in control medium (5 +/- 1) (P < 0.05). These results suggest that oviductal fluid in the estrous bitch maintains sperm motility and induces sperm capacitation.     

Timing of insemination for dairy cows identified in estrus by a radiotelemetric estrus detection system

The optimal time of artificial insemination (AI) was determined from data for 2661 AI in 17 herds utilizing a radiotelemetric system for estrus detection that has the potential for continuous 24-h surveillance to monitor behavioral events associated with estrus. The system consisted of pressure-sensitive radio frequency transmitters affixed over the sacrum region of cows. The activation of the sensor sent a radiotelemetric signal to a microcomputer via a fixed antenna. Cow identification, date, time, and duration of each standing event were recorded in the software program provided with the system. Each farm selected a 3-h interval to AI for cows that were identified in estrus during the previous 24 h. Pregnancy status was determined from data for return to estrus and palpation of the uterus 35 to 75 d following AI. Standing events during estrus averaged (+/- SD) 8.5 +/- 6.6 per cow, and the number of events per estrus across herds averaged from 6.2 +/- 5.1 to 12.8 +/- 9.9 per cow. The duration of estrus ranged from 5.1 +/- 3.8 to 10.6 +/- 6.8 h across herds; the mean was 7.1 +/- 5.4 h. The interval from the first standing event to AI affected the probability of pregnancy; the highest conception rates for AI occurred between 4 and 12 h after the onset of standing activity. The probability of pregnancy was higher for cows > 100 d in milk, exhibiting > 2 standing events during estrus, and inseminated during March, April or May.     

Effect of dehydroepiandrosterone sulfate on uterine artery flow velocity waveforms in term pregnancy

OBJECTIVE: To investigate the interrelation between estrogen synthesis by the fetoplacental unit and uteroplacental hemodynamics in term pregnancy. METHODS: Transvaginal color Doppler flow imaging and pulsed Doppler ultrasonographic assessments were made on ten normal full-term pregnant women before and 3, 5, 10, 30, and 60 minutes after the administration of a 200-mg intravenous dose of dehydroepiandrosterone sulfate (DHAS) in 20 mL of 5% dextrose. Ten normal full-term pregnant women received 20 mL of 5% dextrose as controls. The pulsatility index (PI) values for the uterine artery, heart rate, and mean arterial pressure were recorded. Plasma estradiol (E2) was measured before and 10 minutes after the infusion. RESULTS: In the DHAS group, uterine artery PI decreased from baseline by 26% (P < .05) after 5 minutes, and the mean reduction was 36% (P < .05) after 10 minutes and 15% (P < .05) after 30 minutes. The PI returned to the baseline value 60 minutes later. In the control group, there was no change in uterine artery PI. No change was found in heart rate or mean arterial blood pressure in the control or DHAS groups. The mean plasma E2 increased from 22.3 +/- 6.6 to 56.2 +/- 24.1 ng/mL (P < .05) 10 minutes after the infusion in DHAS subjects, whereas there was no significant change in plasma E2 in the controls. CONCLUSION: Dehydroepiandrosterone sulfate induces a significant decrease in the uterine artery PI, which suggests a possible decrease in uterine vascular impedance in term pregnancy.     

Endometritis in cattle experimentally induced by Chlamydia psittaci

On the day of estrus, eight virgin heifers received intrauterine inoculations of yolk sac propagated Chlamydia psittaci strain BovEnd 11/88 isolated from the uterus of a slaughter cow. All heifers developed purulent vaginal discharge which persisted for 3 to 7 weeks. Chlamydiae or chlamydial antigen were detected in vaginal and uterine discharges of infected animals by culture or Capture ELISA, while other bacterial pathogens were not found. In sera of the chlamydia-infected heifers marked increases in antibody titres against the chlamydial genus-specific LPS-antigen were found by ELISA and complement fixation test. Six heifers were artificially inseminated in 5 successive cycles beginning at the first estrus following intrauterine inoculation. In two of the infected heifers spontaneous healing of endometritis occurred after 5 estrus cycles. Only these animals conceived after the 5th breeding, whereas in the remaining four animals a chlamydia-associated chronic endometritis was recognized as the cause of infertility in the 19th and 26th week p.i. at slaughter. Two control heifers which remained clinically normal after intrauterine exposure to sterile yolk sac-suspensions conceived at the 1st and 2nd service, respectively.     

Effects of space allocation and temperature on periparturient maternal behaviors, steroid concentrations, and piglet growth rates

Preparturient sows were randomly assigned to either a farrowing crate (n = 12) or farrowing pen (n = 12) across cool and hot seasons (with or without drip cooling) to study space allocation and temperature effects on periparturient maternal behaviors, steroid concentrations, and piglet growth rates. Concentrations of estradiol-17 beta (E2 beta), progesterone (P4), and cortisol were quantified in blood collected from surgically implanted vena cava cannulas. Sows were videotaped from 2 h before to 2 h after farrowing. Similar periparturient behaviors were displayed by all sows, regardless of farrowing environment. Sows in pens had lower (P < .05) prepartum P4 concentrations from d -6 to the day preceding farrowing and a reduced (P < .05) piglet birth interval compared with sows in crates (12.95 +/- 1.35 min vs 18.31 +/- 2.21 min, respectively). Additionally, compared with multiparous sows in crates, multiparous sows in pens weaned heavier piglets (P < .05). Estradiol-17 beta concentrations were lower (P < .01) throughout lactation during hot weather for sows with or without drip cooling, averaging 9.30 pg/mL and 8.57 pg/mL, respectively, compared with 18.65 pg/mL for sows during cool weather. This decrease in E2 beta concentration in sows during hot weather was correlated with an extended (P < .05) interval from weaning to first estrus for the sows in hot weather compared to sows during cool weather. Cortisol concentrations decreased progressively during lactation and were not associated with litter weight gains or the interval between weaning and first estrus.     

Conversion of cortisol to cortisone by the human uterus and its reversal in pregnancy

Inter-conversion of cortisol (F) and cortisone (E) was investigated by incubating minced tissue with tritiated cortisol or cortisone and then separating the products by Sephadex LH-20 column chromatography. In non-pregnant subjects conversion of F to E predominated (43.4+/-3.4% vs 0.1+/-0.4% for E to F). In early pregnancy F leads to E decreased and E leads to F rose while at term E leads to F (46.3+/-9.1%) exceeded F leads to E (15.1+/-6.8%). These results were in accord with those obtained by assaying the endogenous concentrations. In non-pregnant subjects the F/E ratio (1.1+/-0.6) was lower than that found in serum (6.3+/-2.2) while at term the uterine F/E (9.0+/-1.8) was similar to that of serum (8.8+/-2.0). These changes resulted in an 8-fold increase in uterine F compared with a 3-fold increase in serum F, while uterine E fell to 1/2 and serum E doubled. Thus, during pregnancy there is a dramatic reversal of the reaction in the uterus in favour of the active hormone. It seems possible that the increase in cortisol thus brought about may play an anti-immune role in uterine wall, the single tissue apart from blood in direct contact with fetal tissue.     

Fertility in estrus-cycling and noncycling virgin heifers and suckled beef cows after induced ovulation

A procedure was developed to either induce or synchronize ovulation in heifers and suckled cows. Beef females were assigned to two breeding programs: 1) two injections of prostaglandin F2alpha (PGF2alpha) given 14 d apart to synchronize estrus (PGF2alpha control; n = 179), with inseminations 12 to 16 h after detected estrus or at 80 h in the absence of estrus, or 2) two injections of PGF2alpha (d -14 and 0) plus 100 microg of GnRH on d -7 when 6 mg of norgestomet was implanted (PGF2alpha/NORG/GnRH treatment; n = 173). Implants were removed 24 h after the second PGF2alpha injection (d +1) and females were inseminated 12 to 16 h after detected estrus until 54 h after PGF2alpha. The remaining cattle were given a second 100-microg GnRH injection 54 h after PGF2alpha and inseminated 18 to 20 h later. Percentages of noncycling females with subsequently elevated progesterone (P4) on d 0 or +1 were not different between treatment groups (20.4 vs 25%), but conception rate was greater (P < .05) in noncycling treated females than in noncycling controls (55 vs 12.8%). Conception rates in cycling (59.2%) and noncycling (62.2%) treated females were similar to those in cycling controls (56.2%) but greater (P = .06) than those in noncycling controls (26.5%). Conception rates in treated females inseminated 12 to 16 h after detected estrus (63.1%) or at one fixed time (58.3%) were similar to those in controls inseminated 12 to 16 h after detected estrus (68.7%). This treatment procedure produced fertility after one timed insemination that was equal to controls inseminated after detected estrus and induced equally fertile ovulations in noncycling heifers and cows.     

Thermal unfolding of dodecameric glutamine synthetase: inhibition of aggregation by urea

Thermal unfolding of dodecameric manganese glutamine synthetase (622,000 M(r)) at pH 7 and approximately 0.02 ionic strength occurs in two observable steps: a small reversible transition (Tm approximately 42 degrees C; delta H approximately equal to 0.9 J/g) followed by a large irreversible transition (Tm approximately 81 degrees C; delta H approximately equal to 23.4 J/g) in which secondary structure is lost and soluble aggregates form. Secondary structure, hydrophobicity, and oligomeric structure of the equilibrium intermediate are the same as for the native protein, whereas some aromatic residues are more exposed. Urea (3 M) destabilizes the dodecamer (with a tertiary structure similar to that without urea at 55 degrees C) and inhibits aggregation accompanying unfolding at < or = 0.2 mg protein/mL. With increasing temperature (30-70 degrees C) or incubation times at 25 degrees C (5-35 h) in 3 M urea, only dodecamer and unfolded monomer are detected. In addition, the loss in enzyme secondary structure is pseudo-first-order (t1/2 = 1,030 s at 20.0 degrees C in 4.5 M urea). Differential scanning calorimetry of the enzyme in 3 M urea shows one endotherm (Tmax approximately 64 degrees C; delta H = 17 +/- 2 J/g). The enthalpy change for dissociation and unfolding agrees with that determined by urea titrations by isothermal calorimetry (delta H = 57 +/- 15 J/g; Zolkiewski M, Nosworthy NJ, Ginsburg A, 1995, Protein Sci 4: 1544-1552), after correcting for the binding of urea to protein sites exposed during unfolding (-42 J/g). Refolding and assembly to active enzyme occurs upon dilution of urea after thermal unfolding.