Hypothalamic expression of genes for appetite regulators and estrogen α, estrogen ß and leptin receptors in obese dams and their fetuses.

Under- and over-nutrition during gestation may influence fetal hypothalamic development resulting in individuals predisposed to adverse health effects. This study examined fetuses from obese and control ewes to determine whether dam obesity alters hypothalamic expression of fetal appetite regulatory genes. A second objective was to contrast the expression of appetite regulatory genes in ewes that become the most obese to those that remained in moderate body condition on the same energy-rich diet. Multiparous, western white-faced ewes were weighed and individually fed 100% (control) or 150% (obese) of National Research Council requirements from day 60 before mating until day 75 of gestation. At day 75 of gestation, fetuses were collected and weighed. Hypothalamic tissue from fetal lambs and dams was collected and frozen for mRNA extraction. Dam obesity (P ≥ 0.16), fetal sex (P ≥ 0.44) or their interaction (P ≥ 0.42) did not affect the relative expression of fetal hypothalamic regulators of appetite, including neuropeptide Y, agouti-related protein, pro-opiomelanocortin, cocaine- and amphetamine-regulated transcript and receptors for leptin. Maternal obesity at day 75 of gestation in ewes did not affect developmental mechanisms responsible for the expression of fetal appetite regulatory genes and would not be expected to predispose offspring to adult-onset obesity through disrupted appetite regulation at this developmental time point. In the ewe, appetite regulatory genes did not differ (P > 0.20) with ewe adiposity; however, expression of estrogen receptor α, but not ß (P = 0.37), in the medial basal hypothalamus was greater (P = 0.04) in obese than in control ewes.

Effects of feeding high-linoleate safflower seeds on postpartum reproduction in beef cows.

Two experiments were conducted to evaluate reproductive responses to supplemental high-linoleate safflower seeds in postpartum beef cows. In Exp. 1, 18 primiparous, crossbred beef cows (411 +/- 24.3 kg of BW) were fed Foxtail millet hay starting 1 d postpartum at 1.68% of BW (DM basis) and a low-fat control (control: 63.7% cracked corn, 33.4% safflower seed meal, and 2.9% liquid molasses; DM basis) at 0.35% of BW (n = 9) or a supplement (linoleate) containing 95.3% cracked high-linoleate (79% 18:2n-6) safflower seeds and 4.7% liquid molasses (DM basis) at 0.23% of BW (n = 9). Beginning 1 d postpartum, blood was collected every 3 d for sera. Cows were slaughtered at 37 +/- 3 d postpartum for collection of hypothalami, anterior pituitary glands, liver, ovarian follicles, and uterine tissue. By 37 +/- 3 d postpartum, dietary treatment did not influence ovarian follicular development (P >or= 0.17), hypophyseal concentrations of LH (P = 0.14), or concentrations of IGF-I in liver (P = 0.15). In contrast, anterior pituitary glands from linoleate cows contained more FSH (P = 0.02) than control cows and linoleate cows had less IGF-I in the medial basal hypothalamus (P = 0.05), preoptic area (P = 0.06), and in follicular fluid (P

Down-regulation of growth signaling pathways linked to a reduced cotyledonary vascularity in placentomes of over-nourished, obese pregnant ewes.

Both protein kinase B (Akt) and extracellular signal-regulated kinase 1/2 (ERK1/2) are down-stream components of the insulin/insulin like growth factor-1 (IGF-1) signaling pathway. AMP-activated protein kinase (AMPK) is known to sensitize cells to insulin/IGF-1 signaling. The objective of this study was to assess the activity of AMPK and its role in the observed down-regulation of insulin/IGF-1 signaling in cotyledonary (COT) arteries supplying the placental component of the ewe placentome. Nonpregnant ewes were randomly assigned to a control (C, 100% of NRC recommendations) or obesogenic (OB, 150% of NRC) diet from 60 days before conception until necropsy on day 75 of gestation (n=5/group) or until lambing (n=5/group). At necropsy on day 75 of gestation, the smallest terminal arteries that entered the COT tissues (0.5-1.0 mm in diameter) were collected for analyses. Fetal weights were approximately 20% greater (P<0.05) on OB than C ewes, but birth weights of lambs were similar across dietary groups. Fetal plasma concentrations of glucose, insulin and IGF-1 were higher (P<0.05) in the blood of fetuses from OB than C ewes. Total AMPK and phosphorylated AMPK at Thr 172 (the active form) were reduced (P<0.05) by 19.7+/-8.4% and 25.9+/-7.7%, respectively in the COT arterial tissues of OB ewes. Total acetyl-CoA carboxylase (ACC), a down-stream target of AMPK, and its phosphorylated form were also reduced (P<0.05) by 32.9+/-9.2% and 45.4+/-14.6%, respectively. The phosphorylation of IRS-1 at Ser 789, a site phosphorylated by AMPK, was 24.5+/-9.0% lower (P<0.05) in COT arteries of OB than C ewes. No alteration in total insulin receptor, total IGF-1 receptor or their phosphorylated forms was observed, down-stream insulin signaling was down-regulated in COT arteries of OB ewes, which may have resulted in the observed decrease in COT vascular development in OB ewes.

A decade of developments in the area of fat supplementation research with beef cattle and sheep.

Supplementing ruminant animal diets with fat has been investigated as a means to influence a variety of physiological processes or to alter fatty acid composition of food products derived from ruminant animals. Several digestion experiments have been conducted with beef cattle and sheep to elucidate the effects of supplemental fat on utilization of other dietary components. Negative associative effects are not likely to be observed in ruminants consuming forage-based diets with supplemental fat at < or = 2% of DMI. Inclusion of supplemental fat at < or = 3% of DM is recommended to obtain the most benefit from the energy contained within the fat and other dietary components in high-forage diets. For ruminants fed high-concentrate diets, supplementing fat at 6% of diet DM is expected to have minimal impacts on utilization of other dietary components. Although there is greater potential to supply the ruminant animal with unsaturated fatty acids from dietary origin if fat is added to high-concentrate diets, incomplete ruminal biohydrogenation of C18 unsaturated fatty acids results in an increase in duodenal flow of 18:1 trans fatty acids regardless of basal diet consumed by the animal. The biohydrogenation intermediate 18:1 trans-11 (trans-vaccenic acid) is the likely precursor to cis-9, trans-11 CLA because the magnitude of increase in CLA content in tissues or milk of ruminants fed fat is much greater than the increase in CLA presented to the small intestine of ruminants fed fat supplements. Duodenal flow of trans-vaccenic acid is also substantially greater than CLA. Increasing unsaturated fatty acids status of ruminants imparts physiological responses that are separate than the energy value of supplemental fat. Manipulating maternal diet to improve unsaturated fatty acid status of the neonate has practical benefits for animals experiencing stress due to exposure to cold environments or conditions which mount an immune response. Supplementing fat to provide an additional 16 to 18 g/d of 18:2n-6 to the small intestine of beef cows for the first 60 to 90 d of lactation will have negative impacts on reproduction and may impair immune function of the suckling calf. Consequences of the suckling animal increasing its intake of unsaturated fatty acids because of manipulation of maternal diet warrants further investigation.

Fatty acid composition of plasma, medial basal hypothalamus, and uterine tissue in primiparous beef cows fed high-linoleate safflower seeds.

The experimental objectives were to evaluate the influence of supplemental high-linoleate safflower seeds on fatty acid concentrations in plasma, medial basal hypothalamus, uterine tissues, and serum 13,14-dihydro-15-keto PGF(2)alpha metabolite (PGFM) in primiparous beef cows during early lactation. Beginning 1 d postpartum, 18 primiparous, crossbred beef cows (411 +/- 24.3 kg of BW) were fed foxtail millet hay at 1.68% of BW (DM basis) and either a low-fat supplement (control: 63.7% cracked corn; 33.4% safflower seed meal; and 2.9% liquid molasses; DM basis) at 0.35% of BW (n = 9) or a supplement (linoleate) containing 95.3% cracked high-linoleate (79% 18:2n-6) safflower seeds and 4.7% liquid molasses (DM basis) at 0.23% of BW (n = 9). Diets were formulated to be isonitrogenous and isocaloric. The linoleate diet contained 5.4% of DMI as fat vs. 1.2% for control. Beginning 1 d postpartum, cattle were bled every 3 d for collection of serum and plasma. Cattle were slaughtered at 37 +/- 3 d postpartum for collection of the medial basal hypothalamus, myometrium, endometrium, caruncular tissue, intercaruncular tissue, and oviduct. Feeding linoleate increased (P = 0.001) plasma concentrations of 18:2n-6, 18:2cis-9 trans-11 and total unsaturated fatty acids; however, 18:1trans-11 did not differ (P = 0.19) between treatments. Concentrations of 20:5n-3 in the medial basal hypothalamus tended (P = 0.10) to be greater for cattle fed linoleate. Concentrations of fatty acids in the oviduct were greater (P < 0.05) than in other uterine tissues. Cows fed linoleate had greater (P = 0.05) concentrations of 18:3n-3 in the endometrium and less (P = 0.06) 18:2cis-9 trans-11 in the myometrium than cows fed the control. Supplemental fat increased (dietary treatment x day postpartum, P = 0.01) concentrations of PGFM in serum more in linoleate than control cows from d 3 to 9 postpartum. Lipid supplementation early in the postpartum period altered the fatty acid composition of medial basal hypothalamus, uterine tissue, and serum concentrations of PGFM. The most novel observation was that the oviduct appeared to be the most sensitive tissue to additional dietary linoleic acid, which could potentially influence fertility.

Influence of short-term fasting during the luteal phase of the estrous cycle on ovarian follicular development during the ensuing proestrus of ewes.

Short-term fasting of mature ewes during diestrus results in increased serum concentrations of progesterone and a delayed pre-ovulatory surge release of LH. To determine if these changes in reproductive hormones influence subsequent follicular development, mature ewes observed in estrus were assigned randomly to control (n=10) or fasted (n=15) groups. Control ewes had ad libitum access to feed, whereas fasted ewes were not fed from day 7 through 11 of their estrous cycle. Daily blood samples were collected from control and fasted ewes throughout the fasting period. Fasting increased (P<0.001) serum concentrations of progesterone (4.4 ng/mL versus 2.7 ng/mL [+/-0.3]). On day 12, all ewes were treated with 10mg of PGF(2alpha) and fasted ewes were returned to ad libitum feed. Ovaries were collected from ewes (n=5 each group) at 0 and 72 h following PGF(2alpha) in control and 0, 72 and 96 h in fasted ewes. Ovaries were weighed and small (< or =2mm), medium (3-4mm), and large (> or =5mm) follicles were enumerated. Total numbers of follicles were less (P<0.001) in fasted than fed ewes (14.6 versus 30.2 [+/-2.2]) at 0 h, but did not differ (P=0.9) when numbers of follicles were compared at similar times before the anticipated LH surge (i.e., at 72 h versus 96 h in control and fasted ewes, respectively). Within follicular size class, numbers of small and medium follicles were decreased (P=0.04) at 0 h in fasted ewes. Numbers of large follicles did not differ (P=1.0) between groups. Although numbers of small and medium ovarian follicles in fasted ewes recovered by 96 h to values comparable to fed ewes at 72 h following PGF(2alpha), serum concentrations of estradiol 17beta (P=0.08) and FSH (P=0.06) tended to be decreased in fasted ewes before the anticipated surge release of LH. Pituitary content of LH and FSH also tended to be lower (P< or =0.09) at 96 h in fasted ewes than at 72 h in control ewes, but did not differ (P> or =0.4) at hour 0 following PGF(2alpha). Hypothalamic and stalk median eminence contents of GnRH were not influenced (P> or =0.2) by fasting at any time period. Fasting during the luteal phase perturbs gonadotropin secretion and may influence fertility by causing a delay in ovarian follicle development.

Effects of postpartum dietary fat and body condition score at parturition on plasma, adipose tissue, and milk fatty acid composition of lactating beef cows.

Two experiments were conducted with lactating Angus x Gelbvieh beef cows to determine the effects of postpartum lipid supplementation, BCS at parturition, and day of lactation on fatty acid profiles in plasma, adipose tissue, and milk. In Exp. 1, 36 pri-miparous cows (488 +/- 10 kg of initial BW; 5.5 +/- 0.02 initial BCS) were given ad libitum access to hay and assigned randomly to a low-fat (control) supplement or supplements with cracked, high-linoleate safflower seeds (linoleate) or cracked, high-oleate safflower seeds (oleate) from d 3 to 90 of lactation. Diets were formulated to be isonitrogenous and isocaloric; safflower seed diets provided 5% of DMI as fat. Plasma and milk samples were collected on d 30, 60, and 90 of lactation. Adipose tissue biopsies were collected near the tail-head region of cows on d 45 and 90 of lactation. In Exp. 2, 3-yr-old cows achieving a BCS of 4 +/- 0.07 (479 +/- 36 kg of BW) or 6 +/- 0.07 (580 +/- 53 kg of BW) at parturition were used in a 2-yr experiment (n = 36/yr). Beginning 3 d postpartum through d 61 of lactation, cows were fed diets similar to those of Exp. 1. Adipose tissue and milk samples were collected on d 30 and 60, and plasma was collected on d 31 and 61 of lactation. Responses to postpartum dietary treatment were comparable in both experiments. Cows fed linoleate and oleate had greater (P < 0.001) total fatty acid concentrations in plasma than cows fed control. Except for 15:1, milk fatty acids with <18 carbons were greatest (P < or = 0.01) for cows fed control, whereas milk from cows fed linoleate had the greatest (P < or = 0.02) 18:1trans-11, 18:2n-6, and cis-9, trans-11 CLA. Milk from cows fed oleate had the greatest (P < 0.001) 18:1cis-9. In Exp. 1, total fatty acid concentrations in adipose tissue samples decreased at d 90 compared with d 45 of lactation, but the fatty acid profile of cow adipose tissue was not affected (P = 0.14 to 0.80) by dietary treatment. In Exp. 2, the percentage of cis-9, trans-11 CLA in adipose tissue of cows with a BCS of 6 decreased (P = 0.001) from d 30 to 60 of lactation. Plasma and milk fatty acid composition reflected alterations in postpartum diet. Less medium-chain fatty acids and more 18-carbon fatty acids in milk were indicative of reduced de novo fatty acid synthesis in the mammary gland of beef cows fed lipid supplements; however, the metabolic demands of lactation prevented the deposition of exogenously derived fatty acids in adipose tissue through d 90 of lactation.

Evaluation of milk somatic cells as a source of mRNA for study of lipogenesis in the mammary gland of lactating beef cows supplemented with dietary high-linoleate safflower seeds.

Our objectives were 2-fold: to determine the effect of dietary linoleate on milk fat composition and on transcript abundance of acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), lipoprotein lipase (LPL), and stearoyl-CoA desaturase (SCD) mRNA in mammary tissue, and to evaluate milk somatic cell mRNA as a source of mammary tissue mRNA for these enzymes. Eighteen primiparous, crossbred beef cows (BW = 411 +/- 24 kg; BCS = 5.25) were offered Foxtail millet hay at 1.68% of BW daily and either a low-fat control (n = 9) or a high-linoleate (79% 18:2n-6), cracked safflower seed supplement (n = 9). Diets were isonitrogenous and isocaloric, and the linoleate diet contained 5.4% of DMI as fat. At slaughter (37 +/- 3 d postpartum), mammary tissue was sampled and immediately frozen in liquid N2 before being stored at -80 degrees C. Milk samples were obtained from the same mammary glands and immediately centrifuged at 1,200 x g to pellet somatic cells. A ribonuclease protection assay was used to quantify the mRNA in the mammary gland and milk somatic cells. Effects of diet, tissue, or their interaction were not observed for ACC (P = 0.28, 0.89, and 0.35, respectively), FAS (P = 0.38, 0.66, and 0.20, respectively), LPL (P = 0.09, 0.15, and 0.43, respectively), or SCD (P = 0.45, 0.19, and 0.29, respectively). Dietary effects on fatty acid profile of the milk fat suggested that linoleate supplementation might have decreased de novo lipogenesis while increasing uptake of dietary fatty acids; this effect was consistent with a trend toward greater LPL mRNA for linoleate-fed cows (P = 0.09). Correlations (r values) between mammary tissue and milk somatic cell data for each mRNA for the low-fat control diet were: ACC, 0.76 (P = 0.02); FAS, 0.69 (P = 0.04); LPL, 0.68 (P = 0.04); and SCD, 0.73 (P = 0.05), and for the linoleate diet were: ACC, 0.85 (P = 0.003); FAS, 0.75 (P = 0.02); LPL, 0.90 (P = 0.001); and SCD, 0.73 (P = 0.03). We conclude that milk somatic cells obtained from lactating beef cows can be used as a source of RNA to study nutritional regulation of mammary gland lipogenesis in cows fed dietary fat supplements.

Effects of body condition score at parturition and postpartum supplemental fat on metabolite and hormone concentrations of beef cows and their suckling calves.

To determine the effects of BCS at parturition and postpartum lipid supplementation on blood metabolite and hormone concentrations, 3-yr-old Angus x Gelbvieh beef cows, which were nutritionally managed to achieve a BCS of 4 +/- 0.07 (479.3 +/- 36.3 kg of BW) or 6 +/- 0.07 (579.6 +/- 53.1 kg of BW) at parturition, were used in a 2-yr experiment (n = 36/yr). Beginning at 3 d postpartum, cows within each BCS were assigned randomly to be fed hay and a low-fat control supplement or lipid supplements with either cracked high-linoleate or high-oleate safflower seeds until d 61 of lactation. The diets were formulated to be isonitrogenous and isocaloric, and the safflower seed supplements were formulated to achieve 5% DMI as fat. On d 31 and 61 of lactation, blood samples were collected preprandially and then hourly postprandially (at 0, 1, 2, 3, and 4 h). Serum insulin (P = 0.27) and glucose (P = 0.64) were not affected by BCS at parturition. The mean concentrations of plasma NEFA (P = 0.08) and beta-hydroxybutyrate (P = 0.08) tended to be greater, and serum IGF-I was greater (P < 0.001) in BCS 6 than BCS 4 cows. Conversely, serum GH was greater (P = 0.003) for BCS 4 cows, indicating that regulation of IGF by GH may have been uncoupled in BCS 4 cows. The postpartum diet did not affect NEFA (P = 0.94), glucose (P = 0.15), IGF-I (P = 0.33), or GH (P = 0.62) concentrations. Oleate-supplemented cows had greater (P = 0.03) serum insulin concentrations, whereas control cows had greater (P = 0.01) plasma beta-hydroxybutyrate concentrations. Concentrations of NEFA (P = 0.05) and glucose (P < 0.001) were greater, and beta-hydroxybutyrate tended (P = 0.07), to be greater at d 3, whereas serum IGF-I was greater (P = 0.003) at d 6 of lactation. Similar concentrations of NEFA, glucose, GH, and IGF-I indicate that the nutritional status of beef cows during early lactation was not influenced by lipid supplementation. However, perturbations of the somatotropic axis in BCS 4 cows indicate that the influence of energy balance and BCS of the cow at parturition on postpartum performance should be considered when making managerial decisions.

Body condition score at parturition and postpartum supplemental fat effects on cow and calf performance.

Three-year-old Angus x Gelbvieh beef cows nutritionally managed to achieve a BCS of 4 +/- 0.07 (479.3 +/- 36.3 kg of BW) or 6 +/- 0.07 (579.6 +/- 53.1 kg of BW) at parturition were used in a 2-yr experiment (n = 36/yr) to determine the effects of prepartum energy balance and postpartum lipid supplementation on cow and calf performance. Beginning 3 d postpartum, cows within each BCS were assigned randomly to be fed hay and a low-fat control supplement or supplements with either high-linoleate cracked safflower seeds or high-oleate cracked safflower seeds until d 60 of lactation. Diets were formulated to be isonitrogenous and isocaloric, and safflower seed supplements were provided to achieve 5% of DMI as fat. Ultrasonic 12th rib fat and LM area were lower (P < 0.001) for cows in BCS 4 compared with BCS 6 cows throughout the study. Cows in BCS 4 at parturition maintained (P = 0.02) condition over the course of the study, whereas cows in BCS 6 lost condition. No differences (P = 0.44 to 0.71) were detected for milk yield, milk energy, milk fat percentage, or milk lactose percentage because of BCS; however, milk protein percentage was less (P = 0.03) for BCS 4 cows. First-service conception rates did not differ (P = 0.22) because of BCS at parturition, but overall pregnancy rate was greater (P = 0.02) in BCS 6 cows. No differences (P = 0.48 to 0.83) were detected in calf birth weight or ADG because of BCS at parturition. Dietary lipid supplementation did not influence (P = 0.23 to 0.96) cow BW change, BCS change, 12th rib fat, LM area, milk yield, milk energy, milk fat percentage, milk lactose percentage, first service conception, overall pregnancy rates, or calf performance. Although cows in BCS of 4 at parturition seemed capable of maintaining BCS during lactation, the overall decrease in pregnancy rate indicates cows should be managed to achieve a BCS >4 before parturition to improve reproductive success.

The incidence of genotypes at codon 171 of the prion protein gene (PRNP) in five breeds of sheep and production traits of ewes associated with those genotypes.

Scrapie is one of several transmissible spongiform encephalopathies of livestock. Disease susceptibility is linked to polymorphisms in the normal prion protein gene that encodes the mammalian prion precursor. Codon 171 of this gene is a major determinant of scrapie susceptibility. Selection for arginine (R) at codon 171 is encouraged by the USDA to decrease the incidence of scrapie. Objectives of this study were to determine the frequency of R allele variants at codon 171 in a sample of sheep from five breeds (Columbia, Hampshire, Rambouillet, Suffolk, and Targhee) and western white-faced commercial ewes and to determine whether the R allele is associated with ewe and lamb production traits. Genotyping was performed on 532 ewes and 901 lambs from the University of Wyoming flock, in addition to 820 rams from 52 sheep producers from Wyoming and surrounding areas, using a DNA mismatch assay that discriminated the R allele from others at codon 171. Genotyping was performed by DNA sequencing on 127 rams representing all breeds, except Hampshire from the USDA Sheep Experiment Station at Dubois, ID. The 171R allele was found in all five breeds and in the commercial western white-faced ewes. Genotype frequencies varied (P < 0.001) by breed in ewe and ram populations. Influence of R-allele frequency on ewe lambing records and individual lamb records was analyzed for Columbia (62, 161, 121), Hampshire (89, 193, 162), Rambouillet (87, 179, 133), Suffolk (67, 178, 161), and commercial sheep (227, 463, 324) for numbers of ewes, total number of ewe production records, and individual lamb records, respectively. Suffolk ewes without the R allele (non-R/non-R) gave birth to more (P or= 0.08) by ewe genotype. Lamb birth and weaning weights were not influenced (P >or= 0.12) by lamb genotype in any of the breeds or in the commercial flock. In this population, ultimate lamb production was only influenced by genotype at codon 171 in the Suffolk flock.

Influence of supplemental cracked high-linoleate or high-oleate safflower seeds on site and extent of digestion in beef cattle.

Our objectives were to evaluate ruminal fermentation patterns, apparent ruminal biohydrogenation, and site and extent of nutrient disappearance in cattle fed supplemental cracked safflower seeds differing in 18 C fatty acid profile. Nine Angus x Gelbvieh heifers (641 +/- 9.6 kg) fitted with ruminal and duodenal cannulas were used in a triplicated 3 x 3 Latin square. Cattle were fed (OM basis) 9.1 kg of bromegrass hay and either 1) 1.8 kg of corn and 0.20 kg of soybean meal (Control); 2) 0.13 kg of soybean meal and 1.5 kg of cracked high-linoleate (67.2% 18:2) safflower seeds (Linoleate); or 3) 1.5 kg of cracked high-oleate (72.7% 18:1) safflower seeds (Oleate). Safflower seed supplements were formulated to provide similar quantities of N and TDN and 5% dietary fat. Single degree of freedom orthogonal contrasts (Control vs. Linoleate and Oleate; Linoleate vs. Oleate) were used to evaluate treatment effects. True ruminal OM and ruminal NDF disappearances (percentage of intake) were greater (P < or =0.02) for Control than Linoleate and Oleate. True ruminal N degradability (% of intake) was not different (P = 0.38) among treatments. Apparent ruminal biohydrogenation of dietary 18:2 was greatest (Linoleate vs. Oleate, P < 0.001) for Linoleate, whereas biohydrogenation of dietary 18:1 was greatest (Linoleate vs. Oleate, P = 0.02) for Oleate. Duodenal flow of 18:0 was least (P < 0.001) for Control but did not differ (P = 0.92) between Oleate and Linoleate. Total flow of unsaturated fatty acid to the duodenum was greatest (P < 0.001) in cattle fed safflower seeds, and was greater with Linoleate (P < 0.001) than with Oleate. Duodenal flow of 18:1 and 18:2 increased (P < 0.001) in Oleate and Linoleate, respectively. Duodenal flow of 18:1trans-11 was greater (P < 0.001) in cattle fed safflower seeds and in Linoleate than in Oleate. Postruminal disappearance of saturated fatty acids was greatest (P < 0.001) for Control; however, postruminal disappearance of total unsaturated fatty acids was greater (P = 0.002) for Linoleate vs. Oleate. Supplemental high-linoleate or high-oleate safflower seeds to cattle fed forage-based diets may negatively affect ruminal OM and fiber disappearance but not N disappearance. Provision of supplemental fat in the form of safflower seeds that are high in linoleic acid increased intestinal supply and postruminal disappearance of unsaturated fatty acids, indicating that the fatty acids apparently available for metabolism are affected by dietary fat source.

Effects of feed restriction on reproductive and metabolic hormones in ewes.

The goal of this study was to determine the effects of short-term feed withdrawal on reproductive and metabolic hormones during the luteal phase of the estrous cycle in mature ewes. Mature ewes observed in estrus were assigned randomly to control and fasted groups (n = 10 per group Trials 1 and 2). For Trials 1 and 2, control ewes had ad libitum access to feed, whereas fasted ewes were not fed from d 7 through 11 of their estrous cycle; on d 12, all ewes were treated with 10 mg of PGF2alpha, and fasted ewes were gvien ad libitum access to feed. For Trial 1, blood samples were collected daily through fasting and at 2-h intervals following PGF2alpha for 72 h. Serum concentrations of insulin (P < or = 0.002) and IGF-I (P < or = 0.01), but not GH (P > or = 0.60), were decreased during fasting compared with fed ewes. Serum concentrations of 29 (P = 0.02) and 34 kDa (P = 0.04) IGFBP were greater in fasted ewes at 96 h after initiation of fasting than in control ewes. Two control and four fasted ewes in Trial 1 did not exhibit a preovulatory surge release of LH by 72 h. Therefore, Trial 2 was conducted so that the timing of the LH surge could be predicted following the collection of blood samples at 2-h intervals for 112 h and then at 6-h intervals until 178 h following PGF2alpha administration and realimentation. The magnitude of the preovulatory LH surge in Trial 2 was decreased (P = 0.009) and delayed (P = 0.04), and serum concentrations of estradiol were diminished (P < or = 0.03) 12 h before the LH surge in fasted ewes. Ovulation rates were not influenced (P > or = 0.32) by fasting in Trials 1 and 2. Serum concentrations of progesterone in both Trials 1 and 2 were, however, greater (P < 0.001) in fasted than in control ewes. A third trial with ovariectomized ewes was conducted to determine whether the increased serum concentrations of progesterone observed in fasted ewes during Trials 1 and 2 were ovarian-derived. Ovariectomized ewes were implanted with progesterone-containing intravaginal implants and allotted to control (n = 5) or fasted (n = 5) treatment groups and fed as described for Trials 1 and 2. Similar to intact ewes, serum concentrations of progesterone were approximately twofold greater (P < 0.001) in fasted than in control implanted ovariectomized ewes. In summary, feed withdrawal for 5 d during the luteal phase of the estrous cycle increased serum concentrations of progesterone and evoked endocrine changes that could perturb the subsequent estrous cycle.

Effects of supplementation with high linoleic or oleic cracked safflower seeds on postpartum reproduction and calf performance of primiparous beef heifers.

Primiparous Angus x Gelbvieh (n = 36) rotationally crossed beef cows (initial BW = 487.9 +/- 10.5 kg, body condition score = 5.5 +/- 0.02) were utilized to determine effects of supplemental safflower seeds high in linoleic (76% 18:2) or oleic (72% 18:1) acid on cow BW change, body condition score, milk production and composition, calf weight gain, cow serum metabolites, and metabolic hormones. On d 3 postpartum, cows were randomly assigned to one of three isonitrogenous dietary supplements with equal total quantity of TDN: corn-soybean control supplement (n = 12); high-linoleate safflower seeds (n = 12); or high-oleate safflower seeds (n = 12). Safflower-seed supplements were formulated to provide 5% DMI as fat. Supplements were individually fed from d 3 postpartum through 90 d postpartum. Cows had ad libitum access to native grass hay (7.8% CP), trace-mineralized salt, and water. Date of parturition was evenly distributed across treatments with all cows calving within 14 +/- 0.8 d. There were no differences (P = 0.65) in total OM intake among treatments. Although cow BW change did not differ (P = 0.33) by treatment, supplementation influenced cow body condition score (P = 0.02) with linoleate-supple-mented cows in higher (P = 0.005) condition overall than oleate-supplemented cows (5.1 +/- 0.06 vs 4.9 +/- 0.06). Twenty-four-hour milk production did not differ (P = 0.68) among treatments. Percentage milk fat was not different at d 30; however, at d 60 and d 90 percentage milk fat was greater (P ( 0.05) in control and oleate-supplemented cows than in linoleate-supplemented cows. Calf BW gains (P = 0.27) and adjusted 205-d weights (P = 0.48) were not affected by supplement treatment. Supplementation did not influence serum concentrations of glucose (P = 0.38), NEFA (P = 0.61), GH (P = 0.29), IGF-I (P = 0.81), insulin (P = 0.26), or IGF-I binding proteins (P > or = 0.11). Days to conception did not differ (P = 0.40) among treatments. Although overall productivity of the primiparous cows and their calves was not altered by safflower-seed supplementation, differential effects were noted between supplements. Oleate supplementation increased percentage milk fat at d 60, and cow body condition score was lower than in linoleate-supplemented cows. Linoleate-supplemented cows had greater body condition scores by 90 d postpartum than either corn-soybean- or oleatesupplemented cows.

Feeding lambs high-oleate or high-linoleate safflower seeds differentially influences carcass fatty acid composition.

Our objective was to determine effects of dietary high-oleate (Oleate; 76% 18:1) or high-linoleate (Linoleate; 78% 18:2) safflower seeds on fatty acids in muscle and adipose tissue of feedlot lambs. White-faced ewe lambs (n = 36) were fed a beet pulp, oat hay, and soybean meal basal diet (Control), blocked by BW, and allotted randomly to dietary treatments. Cracked safflower seeds were used in isocaloric and isonitrogenous replacement of beet pulp, oat hay, and soybean meal so that Oleate and Linoleate diets contained 5.0% additional fat. Fatty acids were determined in semitendinosus, longissimus dorsi (longissimus), and adipose tissue from the tail head (tailhead adipose tissue), adjacent to the 12th rib (s.c. adipose tissue), and kidney and pelvic fat (KPH adipose tissue) depots. Fatty acid data were analyzed within muscle and adipose tissue as a split-block design. Single degree of freedom orthogonal contrasts were used to compare treatment effects. Average daily gain, feed efficiency, and carcass characteristics did not differ (P = 0.15 to 0.96) across dietary treatments. Adipose tissue saturated fatty acids were greater (P = 0.04) for Controls but were not different (P = 0.36) in muscle. Trans-vaccenic acid (18:1(trans-11)) increased (P < 0.0001) with safflower supplementation and was greater (P < 0.0001) in Linoleate than in Oleate for both tissue types. Linoleate lamb had greater (P < 0.0001) PUFA than Oleate lamb in muscle and adipose tissue. Conjugated linoleic acids (CLA; cis-9, trans-11 and trans-10, cis-12) were greater (P < 0.0001) in muscle and adipose tissue of lambs fed safflower seeds. Lambs fed Linoleate had greater (P < 0.0001) CLA in adipose tissue and muscle than lambs fed Oleate. Saturated fatty acids were greater (P < 0.0001) in s.c. adipose tissue than in tailhead adipose tissue. Mono- and polyunsaturated fatty acids were greater (P < 0.0001) in tailhead adipose tissue than in s.c. adipose tissue. Weight percentages of 18:1(trans-11) ranked tailhead adipose tissue = KPH adipose tissue > s.c. adipose tissue and semitendinosus > longissimus, whereas CLA ranked tailhead adipose tissue > s.c. adipose tissue > KPH adipose tissue and semitendinosus > longissimus. Feeding mono- and polyunsaturated fatty acids increased tissue 18:1(trans-11) and CLA, which is a favorable change in regard to current human dietary guidelines.

Insulin-like growth factor binding proteins in the bovine anterior pituitary.

Insulin-like growth factor binding proteins (IGFBPs) were characterized in bovine anterior pituitary tissue, pituitary conditioned media, and serum collected during the preovulatory and early luteal phases of the estrous cycle. Effects of in vitro treatments of pituitaries with luteinizing hormone-releasing hormone (LHRH), estradiol, and progesterone on IGFBP secretion were also evaluated. Predominant IGFBPs detected in anterior pituitary tissue by immunoprecipitation, ligand blotting, and Northern blotting were IGFBP-5 (29 kDa), IGFBP-2 (32 kDa), and IGFBP-3 (36 and 39 kDa doublet). Conditioned culture media contained IGFBP-5, a slightly larger form of IGFBP-2 (33 kDa), the 36- and 39-kDa forms of IGFBP-3, and a more extensively glycosylated form of IGFBP-3 (44 kDa). In serum, IGFBP-5 was not readily detected, and IGFBP-3 (40- and 44-kDa doublet) and IGFBP-2 (34 kDa) were larger than in pituitary tissue. Levels of IGFBP-2, -3, and -5 in pituitary tissue decreased during the preovulatory period and were lowest in the early luteal phase. Treatment with LHRH increased IGFBP-2 levels in media twofold. Estradiol or progesterone did not alter IGFBP secretion in vitro. Predominant IGFBPs produced and released by anterior pituitary tissue were IGFBP-2, -3 and -5. The activity of IGFBPs fluctuates in the pituitary in association with changes in stage of estrous cycle, implicating IGFBPs as potential regulators of gonadotrope function.

Fos-like immunoreactivity in brain regions of domestic rams following exposure to rams or ewes.

Limbic and basal forebrain-hypothalamic regions from male sheep differing in sexual performance were quantified for fos-like immunoreactivity. Rams classified as high-sexually performing (HP), low-sexually performing (LP), and male-oriented (MO) received noncontact sensory stimulation from either ewes in estrus (HP, n=5; LP, n=4; MO, n=4) or other males (HP, n=5; LP, n=4; MO, n=5) for a 4-h period on each of 3 consecutive days. Following exposure to stimulus animals on the third day, rams were euthanized and their brains were perfused with a 1% paraformaldehyde/1.5% glutaraldehyde solution and sections were analyzed for fos-like immunoreactivity. Brain regions analyzed were the medial amygdala (meAMY), medial preoptic area (mPOA), bed nucleus of the stria terminalis (BNST), and ventromedial hypothalamic nucleus (VMH). Fos-like immunoreactivity differed between groups in the mPOA and BNST but not in the meAMY or VMH. LP rams exposed to estrous ewes had more (P<.05) neurons staining positive for fos and fos-related antigens (FRA) in the mPOA and BNST than LP rams exposed to other rams or MO rams exposed to either estrous ewes or other rams. Numbers of neurons staining positive for FRA in the mPOA and BNST of LP rams exposed to estrous ewes, however, were not different (P>.05) from HP rams exposed to either estrous ewes or other rams. The similar fos-like immunoreactivity in areas important for the display of sexual behavior in HP and LP rams may reflect similar sensory input in these two groups of rams; however, LP rams, in contrast to HP rams, do not appear to respond similarly to the same sensory stimulus.

Low-sexually performing rams but not male-oriented rams can be discriminated by cell size in the amygdala and preoptic area: a morphometric study.

Brain regions of male sheep behaviorally classified as high-sexually performing (n=10), low-sexually performing (n=8) or male-oriented (n=9) were examined to determine if differences in reproductive behavior were associated with differences in density or sizes of neurons. High-sexually performing rams actively mounted estrous ewes, low-sexually performing rams failed to mount or had long latencies to mounting estrous ewes, and male-oriented rams mounted other rams in preference to ewes in estrus. Cell densities and sizes were quantified in Nissl stained sections through the medial amygdala (meAMY), preoptic area (POA), bed nucleus of the stria terminalis (BNST), ventromedial hypothalamic nucleus (VMH), lateral geniculate nucleus (LG) and medial geniculate nucleus (MG). Multivariate discriminant analysis based on soma sizes within nuclei of known importance for reproductive behavior and/or gonadotropin release (meAMY, POA, BNST and VMH) discriminated (Wilks Lambda P<0.05) low-performing rams from high-performing and male-oriented rams, but did not discriminate (Wilks Lambda P=0.14) between high-performing and male-oriented rams. Cell size in the parvocellular and magnocellular layers of the LG along with cells of the MG, structures without a specific role in reproduction, did not discriminate any of the three behaviorally defined groups of rams (Wilks Lambda P=0.57). Density of cells present in structures important for the display of reproductive behavior (POA, meAMY, BNST) and/or gonadotropin release (POA, VMH) had no discriminating power nor did density of cells in structures important for the processing of visual (LG) or auditory (MG) stimuli. In conclusion, significant differences in sizes of cells located within nuclei that are specifically important for the display of male reproductive behavior were found in low-sexually performing rams compared to high-sexually performing and male-oriented rams. These differences may result from neuron development in utero or occur later as a consequence of endocrine factors or behavioral experience. Neuronal cell size is a critical variable that determines excitability to synaptic inputs because cell surface area varies exponentially with cell diameter. Relatively small differences in neuron diameter could relate to functionally important differences in neuronal excitability.

Gonadal function, sexual behavior, feedlot performance, and carcass traits of ram lambs actively immunized against GnRH.

The effect of active immunization against GnRH on production, carcass, and behavioral traits was examined in ram lambs fed to a uniform slaughter weight. Lambs (initial BW = 32.6+/-1 kg) were stratified by BW and assigned at random to one of four treatment groups (n = 12 lambs/group). Lambs were untreated, castrated, or actively immunized against GnRH using a GnRH-keyhole limpet hemocyanin conjugate (1 mg) emulsified with either Freund's complete adjuvant (FCA) or another oil-based adjuvant (ISA). Animals were housed individually and slaughtered at 58 kg BW. Immunoneutralization of GnRH reduced (P < .05) testes weight and the concentration of testosterone in serum at slaughter. Suppression of testicular size and function was most clearly evident in animals immunized using FCA. Final anti-GnRH titer was also highest in lambs immunized using FCA. Several measures of sexual behavior (frequency of mounts and ejaculations) were also reduced (P < .05) in animals immunized using FCA. The duration of the feeding period was greater (P < .05) for castrated lambs than for untreated lambs, and intermediate feeding periods were required for FCA and ISA lambs. Average daily gain was greater (P < .05) in untreated than in castrated, FCA, or ISA lambs. Similarly, feed efficiency for untreated lambs was greater (P < .05) than for castrated, FCA, or ISA lambs, but feed efficiency did not differ among castrated, FCA, or ISA lambs. Longissimus muscle area, lean and bone maturity, overall quality, muscling score, flank streaking, and color of fat did not differ among treatments. Intact, FCA, and ISA lambs had more (P < .05) desirable yield grades, less (P < .05) backfat, and less (P < .05) marbling than castrated lambs. In summary, immunization against GnRH decreased testicular weight and reduced (P < .05) feedlot performance and sexual behavior to levels comparable to those of castrated males. Partitioning of nutrients for growth and deposition of fat, however, seems to differ among immunologically castrated and physically castrated lambs. This difference in nutrient partitioning may be due to residual testicular activity in immunized lambs.

Effects of supplemental protein type on productivity of primiparous beef cows.

Effects of supplemental degradable (DIP) and undegradable (UIP) intake protein on forage intake, BW change, body condition score (BCS), postpartum interval to first estrus, conception rate, milk production and composition, serum metabolites and metabolic hormones, and calf gain were determined using 36 primiparous Gelbvieh x Angus rotationally crossed beef cows. On d 3 postpartum, cows (average initial BW = 495 +/- 10 kg and BCS = 5.5 +/- 0.1) were randomly assigned to one of three dietary supplements (12 cows/treatment). Date of parturition was evenly distributed across treatment (average span of calving date among treatments = 2.4 +/- 2.5 d). Individually fed (d 3 through 120 postpartum) dietary supplements were 0.82 kg of corn and 0.23 kg of soybean meal per day (DIP), the DIP + 0.12 kg of blood meal and 0.13 kg of corn gluten meal per day (DIP + UIP), and 0.82 kg of corn, 0.07 kg of blood meal, and 0.08 kg of corn gluten meal per day in an isonitrogenous replacement of soybean meal (UIP IsoN). Cows had ad libitum access to native grass hay (8.5% CP) and trace-mineralized salt. Total OM intake was greater (P = 0.06) for DIP + UIP than UIP IsoN cows. At 30 d postpartum, DIP + UIP cows produced more milk than UIP IsoN, with DIP being intermediate; however, at 60 d postpartum, DIP + UIP and DIP cows were not different, but both had greater milk production than UIP IsoN (treatment x day interaction; P = 0.08). A treatment x day interaction (P = 0.06) for BCS resulted from DIP + UIP cows having the greatest BCS at 60, 90, and 120 d d postpartum and DIP having greater BCS than UIP IsoN cows only on d 60 postpartum. Serum insulin concentrations were highest (treatment x day interaction; P = 0.09) for DIP + UIP cows at 30 d postpartum but did not differ among treatment thereafter. Serum insulin-like growth factor-binding protein (IGFBP)-2 (34 kDa) and -3 (40 and 44 kDa) were greatest (P < 0.0003) for DIP cows. Serum urea-N concentrations were greater (P < 0.01) in DIP + UIP cows than in either DIP or UIP IsoN cows. However, postpartum interval to first estrus, conception rate, and calf weaning weights were unaffected (P = 0.35, 0.42, and 0.64, respectively) by treatment. Although UIP in addition to or in replacement of DIP affected milk production and blood metabolites, the productivity of these primiparous beef cows was not altered. Thus, the type of supplemental protein does not seem to influence productivity of primiparous beef cows in production systems with conditions similar to our experimental conditions.