Effects of dietary protein and growth hormone-releasing peptide (GHRP-2) on plasma IGF-1 and IGFBPs in Holstein steers

This study was conduct to determine the influence of dietary protein on the response of plasma insulin-like growth factor-1 (IGF-1) and insulin-like growth factor binding proteins (IGFBPs) to exogenous growth hormone releasing peptide-2 (GHRP-2 or KP 102) in Holstein steers. Eight 16-month-old Holstein steers were grouped by liveweight to two feeding treatments; high protein (HP; CP 1.38 kg/day and TDN 4.5 kg/day DM intake, n=4) or low protein (LP; CP 0.66 kg/day and TDN 4.42 kg/day DM intake, n=4). The experiment was a single reverse design whereby each group was injected twice daily with GHRP-2 (12.5 microg/kg body weight (BW)/day) or saline solution into the jugular vein for a 6-day period. Plasma IGF-1 in the HP group were higher than in the LP group (P<0.05), but plasma 34 kDa IGFBP-2 was lower in the HP than the LP group (P<0.05). The amplitude of the maximum growth hormone (GH) peaks responding to GHRP-2 injection were higher at day 1 than at day 6 of saline or GHRP-2 treatment in both LP and HP steers (P<0.05). The area under the GH response curve for 180 min after the GHRP-2 injection was not significantly different between the LP and the HP groups at days 1 and 6. A response in plasma IGF-1 concentration to GHRP-2 treatment in the HP group was observed at day 1 (198.9+/-18.1 ng/ml), day 2 (195.2+/-21.1 ng/ml) and day 6 (201.3+/-14.8 ng/ml) (P<0.05). No increase in plasma IGF-1 was observed from GHRP-2 administration in the LP group. Although the response of plasma IGF-1 concentration to GHRP-2 administration was increased in the HP group (P<0.05), there was no apparent effect of GHRP-2 treatment on plasma 38-43 kDa IGFBP-3 and 34 kDa IGFBP-2 at days 2 and 6 of treatment. In conclusion, it is proposed that the 34 kDa IGFBP-2 is sensitive to dietary protein level and may play an important role in the regulation of circulating IGF-1 in ruminant. In addition, increased plasma IGF-1 concentration observed in the HP group in response to the GHRP-2 treatment did not appear to affect plasma IGFBPs.     

Feed restriction in young bulls alters the onset of puberty in relationship with plasma insulin-like growth factor-I (IGF-I) and IGF-binding proteins

The objectives of this study were to evaluate the effect of feed restriction and re-alimentation on the onset of puberty and IGF status in peripubertal male calves and to compare the radioimmunoassay (RIA) and western ligand blotting (WLB) methods for bovine IGFBP-2. Twelve prepubertal 290 d-old Belgian Blue bulls (mean weight: +/- 290 kg) were randomly assigned in three groups: a control group (NG; n = 4) receiving a classic fattening diet to induce "normal" growth (1.48 kg/d), a feed restricted group (RG; n = 4) to obtain reduced growth (0.50 kg/d) and, a severely restricted group (SG; n = 4) to nearly stop growth (0.08 kg/d). The feed restriction period was maintained over a period of 114 d. After the period of differential feeding, all animals received the control feed regime over a period of 100 d. Blood samples were collected at fortnightly intervals. Circulating IGF-I was measured by RIA whereas plasma IGFBPs was evaluated by WLB; IGFBP-2 was additionally quantified by RIA procedure. At the beginning of the trial, IGF-I levels were low (<100 ng/ml) and similar in the three groups in accordance with prepubertal status. In the NG group, a progressive rise in IGF-I was observed from Day 42 to Day 142 whereas in the RG and SG groups, IGF-I levels did not change until the experimental restriction period ended. The delay of the rise in plasma IGF-I was longer for the SG group, IGF-I remained low until 2 wk after the end of the period of restricted feeding. Surprisingly, although differences were detected for IGF-I levels between the three groups, the IGFBP-2 and -3 data, evaluated by WLB could only discriminate between NG and SG group and not between NG and RG. However, by using a RIA method, an IGFBP-2 decrease was observed in the NG group coincident with increasing IGF-I levels. For both RG and SG groups, IGFBP-2 levels remained high throughout the feed restriction period whereas plasma IGFBP-2 levels declined upon feeding in both groups. During this feed restriction period, IGFBP-2 was significantly lower in NG than in RG or SG groups. Moreover, SG group animals had higher levels in plasma IGFBP-2 than RG animals. In conclusion, puberty is characterized by developmental changes in plasma IGF-I and IGFBPs that were altered by feed restriction. Moreover, RIA evaluation of plasma IGFBP-2 is able to better reflect group differences than WLB.     

Acute handling disturbance modulates plasma insulin-like growth factor binding proteins in rainbow trout (Oncorhynchus mykiss)

The effects of acute stressor exposure on proximal (growth hormone [GH]) and distal (insulin-like growth factor-I [IFG-I] and insulin-like growth factor-binding proteins [IFGBPs]) components of the somatotropic axis are poorly understood in finfish. Rainbow trout (Oncorhynchus mykiss) were exposed to a 5-min handling disturbance to mimic an acute stressor episode, and levels of plasma GH, IGF-I, and IGFBPs at 0, 1, 4, and 24 h post-stressor exposure were measured. An unstressed group was also sampled at the same clock times (09:00, 10:00, 13:00, and 08:00 [the following day]) as acute stress sampling to determine temporal changes in the above somatotropic axis components. The acute stressor transiently elevated plasma cortisol and glucose levels at 1 and 4 h post-stressor exposure, whereas no changes were seen in the unstressed group. Plasma GH levels were not affected by handling stress or sampling time in the unstressed animals. Plasma IGF-I levels were significantly depressed at 1 and 4 h post-stressor exposure, but no discernible temporal pattern was seen in the unstressed animals. Using a western ligand blotting technique, we detected plasma IGFBPs of 21, 32, 42, and 50 kDa in size. The plasma levels of the lower-molecular-weight IGFBPs (21 and 32 kDa) were unaffected by handling stressor, nor were there any discernible temporal patterns in the unstressed animals. By contrast, the higher-molecular-weight IGFBPs (42 and 50 kDa) were affected by stress or time of sampling. Levels of the 42-kDa IGFBP levels significantly decreased over the sampling period in unstressed control animals, but this temporal drop was eliminated in stressed animals. Levels of the 50-kDa IGFBPs also decreased significantly over the sampling time in unstressed trout, whereas handling disturbance transiently increased levels of this IGFBP at 1 h but not at 4 and 24 h post-stressor exposure compared with the control group. Overall, our results suggest that acute stress adaptation involves modulation of plasma IGF-1 and high-molecular-mass IGFBP levels (42 and 50 kDa) in rainbow trout.     

Oral administration of peptidergic growth hormone (GH) secretagogue KP102 stimulates GH release in goats

To assess the oral activity of KP102 (also known GHRP-2) on growth hormone (GH) release in ruminant animals, 5 or 10 mg/kg body weight (BW) of KP102 dissolved in saline was orally administered twice at 2 hr-intervals to either 1- or 3-mo-old goats (n = 5-6). Plasma GH concentrations in the 1-mo-old goats were elevated at 15 min after the first administration of both 5 and 10 mg/kg BW of KP102. Significant elevation of GH concentrations continued until 180 min after 10 mg/kg BW of KP102, whereas the elevated GH levels after the administrations of 5 mg/kg BW of KP102 subsided to basal concentrations within 90 min. The second administration of 10 mg/kg BW of KP102 failed to elevate the GH concentration, but 5 mg/kg BW of KP102 abruptly stimulated GH release. Plasma GH concentrations in the 3-mo-old goats were also significantly elevated after the administration of both 5 and 10 mg/kg BW of KP102. The plasma GH responses to 5 and 10 mg/kg BW of KP102 were almost identical. The elevated GH levels after the first administration of KP102 tended to be maintained throughout the experiment, and a transient increase in plasma GH levels was observed after the second administration. However, the stimulatory effect of KP102 on GH release in the 3-mo-old goats was small and less abrupt than that in the 1-mo-old goats. The concentrations of insulin-like growth factor-I were not increased by KP102 during the brief sampling periods used in this experiment. These results show that the oral administration of the peptidergic GH secretogogue KP102 stimulates GH release in a ruminant species, and that the oral activity of KP102 on GH release is modified by the age.     

Effects of a four-day hyperinsulinemic-euglycemic clamp in early and mid-lactation dairy cows on plasma concentrations of metabolites, hormones, and binding proteins.

The effects of insulin, using a 4 d hyperinsulinemic-euglycemic clamp, on plasma concentrations of hormone, metabolites, and binding proteins were evaluated in four Holstein dairy cows during wk 4 and 17 of lactation. Insulin was infused at 1 microg/kg/hr for 96 hr during the clamp period. Compared with the pre-clamp period, plasma insulin concentrations increased 7-fold and 4-fold during the clamp periods in early and mid-lactation, respectively. The total amount of glucose infused was higher (P < 0.05) during the clamp in early lactation. The clamp decreased plasma concentrations of non-esterified fatty acids (P < 0.001) during early lactation while differences in mid-lactation were minor. The clamp also decreased plasma concentration of beta-hydroxybutyrate (P < 0.001), plasma urea nitrogen (P < 0.001), and true protein (P < 0.01) although the patterns of decline differed between early and mid-lactation. Growth hormone (GH) concentrations decreased (P < 0.001) and insulin-like growth factor-1 (IGF-1) increased (P < 0.01) during the clamp period suggesting a direct effect of insulin on the un-coupling of the GH/IGF-1 axis. Levels of IGF binding protein-2 (IGFBP-2) decreased (P < 0.01) during the clamp period. The relative proportion of IGFBP-2 decreased (P < 0.001) and that of IGFBP-3 increased (P < 0.001) during the clamp period. There were no interactions between the clamp period and stage of lactation on GH, IGF-1, or IGFBPs. Overall, most plasma variables measured were affected in the same way during the two clamps, but the pattern of change often varied with stage of lactation.     

Alterations of serum insulin-like growth factor-I (IGF-I) and IGF-binding proteins (IGFBPs) in swine infected with the protozoan parasite Sarcocystis miescheriana.

The effects of a Sarcocystis miescheriana infection on insulin-like growth factor-I (IGF-I) and insulin-like growth factor binding proteins (IGFBPs) were investigated to determine possible mechanisms of growth retardation in growing pigs. Sixteen pigs averaging 14 kg body weight were divided into 4 groups of 4 pigs each and infected either with 0.5, 1.0, or 3.0 × 10⁶ sporocysts of S. miescheriana. Four pigs were retained as non-infected controls; however, they became serologically positive during the course of the infection. Effects also were investigated in 2 groups of 3 pregnant sows. One group was infected with 0.5 × 10⁶ sporocysts and the other group was retained as uninfected controls. Body weights of infected growing pigs were depressed as compared to controls following the acute phase 15 d after infection (dai). Serum concentrations of IGF-I dropped significantly (p < 0.05) during the acute phase of infection in all infected groups of growing pigs. Conversely, the amounts of unsaturated serum IGFBPs were elevated significantly (p < 0.05) during the acute phase of infection. Specifically, serum concentrations of IGFBP-1, IGFBP-2, and IGFBP-4 were elevated at this time, as determined by ligand blot analysis. There was no association between growth factor alterations and tissue damage as measured by serum creatinine kinase and aspartate aminotransferase levels. The extent of effects in growing pigs was related to the amount of the original parasite inoculum.

During the acute phase of infection 2 of 3 pregnant sows aborted. The third sow went to term, but piglets were stillborn or died within 24 hr. Compared to uninfected controls, serum concentrations of IGF-I in infected pregnant sows were depressed during and after the acute phase of the infection. Levels of unsaturated serum IGFBPs in pregnant sows were not affected.

These data suggest that decreased IGF-I levels and/or elevated levels of specific forms of IGFBPs may be a mechanism by which growth is affected in feeder pigs infected with S. miescheriana.     

Predicting growth and performance of Holstein steers

Refinements and additions were made to the growing cattle portion of the model of Fox et al. (1988) for predicting growth of Holstein steers. Based on observations of animal weight, DM intake (DMI), ADG, feed per gain (FPG) and diet ME, the modified model was tested on a database that included 299 feeding periods with Holstein steers fed in research trials over a period of 15 yr at Cornell University, the University of Minnesota and the University of Wisconsin. The modified model accounted for 93, 56 and 68% of the variation in DM intake, daily gain and feed required per unit gain, respectively. Averaged over all 299 periods in the database, the error in predicting DM intake, daily gain and feed required per unit gain was 1%, 4% and 2%, respectively. Analysis of residuals indicated that ME level did not influence the difference between predicted and observed values for DMI, ADG and FPG. Residuals analysis indicated that BW did not influence the difference between predicted and observed values for ADG and FPG but DMI of heavier cattle was overpredicted (P less than .01). These results indicated that the 8% increase in DMI for cattle over 318 kg was not warranted for the Holstein steers in this database, which were predominantly less than 15 mo at slaughter. The results of the validation indicate that this model accurately predicted performance of Holstein steers.     

Insulin-like growth factors and IGF-binding proteins in bovine seminal plasma.

Insulin-like growth factor-I (IGF-I) is an important factor for germ cell development and maturation of spermatozoa. Actions of IGFs are modulated by IGF-binding proteins (IGFBPs) that may, depending on their concentration and site of expression, inhibit or enhance effects of IGF-I. We characterized IGFs and IGFBPs in seminal plasma from bulls routinely used for artificial insemination (AI) and from bulls producing poor-quality semen (low mass and individual motility of spermatozoa). IGFs were measured by specific radioimmunoassay in 22 samples of seminal plasma from nine different AI bulls with high (> 76.8%), average (72.8-73.4%), or low (< 69.5%) nonreturn rate (NRR). IGF-I and IGF-II levels were 144 +/- 9 ng/ml (mean +/- SE; range, 79-238 ng/ml) and 144 +/- 10 ng/ml (range, 55-221 ng/ml), respectively, and did not correlate with NRRs. IGF-I concentrations in seminal plasma from bulls producing poor-quality semen (n = 10) were significantly (P < 0.05) greater (194 +/- 26 ng/ml; range, 94-370 ng/ml), whereas IGF-II levels were significantly (P < 0.05) lower (93 +/- 17 ng/ml; range, 38-183 ng/ml) than in AI bulls. Ligand blot analysis of seminal plasma for IGFBPs revealed the presence of a 38-/45-kDa doublet band and a 30-kDa IGFBP. These IGFBPs were identified as IGFBP-3 and IGFBP-5, respectively, by immunoprecipitation using specific antibodies. In addition, a low amount of IGFBP-4 was detected in bovine seminal plasma by immunoprecipitation. There was a marked difference in the activity of IGFBPs between individual bulls, with a relatively small within-bull variance. The differences in IGFBP activities did not correlate with the fertilization capacity of the bulls in vivo or in vitro nor with immunoreactive IGF-I and IGF-II levels in seminal plasma. Our results demonstrate the presence of IGFBPs in bovine seminal plasma. In contrast to human seminal plasma, high activity of IGFBP-3 was detected in seminal plasma of some bulls, suggesting species-specific regulation of IGFBP activity by proteases.     

Effect of feeding wood kraft pulp on the growth performance,feed digestibility,blood components,and rumen fermentation in Japanese Black fattening steers

This study aimed to examine the effects of feeding kraft pulp (KP) on the growth performance, feed digestibility, and rumen fermentation of Japanese Black fattening steers. Ten Japanese Black fattening steers (aged 26 months) were randomly divided into control and KP groups. The control group (n = 5) was fed concentrate feed without KP, and the KP group (n = 5) was fed concentrate feed containing 10% KP. Both the groups were provided rice straw as roughage. The experiment was conducted over a period of 12 weeks. There was no significant difference in dry matter intake, daily body weight gain, and nutrient digestibility between both groups. No difference was observed in the ruminal concentrations of volatile fatty acids among the groups. At weeks 8 and 12 after the onset of the experiment, the acetate‐to‐propionate ratio in the ruminal fluid of the KP group was significantly higher than that of the control group. The average daily pH of ruminal fluid and activity of ruminal lipopolysaccharide did not differ between the groups. Our results suggested that the growth performance and feed digestibility in the Japanese Black fattening steers were not influenced by replacing concentrate feed with KP.     

Possible role of growth hormone, IGFs, and IGF-binding proteins in the regulation of ovarian function in large farm animals.

The aim of the study and short review was to present evidence that growth hormone (GH), locally produced insulin-like growth factors (IGFs), and IGF-binding proteins (IGFBPs) may have an important role in the control of ovarian function. There is clear evidence for a distinct GH-receptor mRNA expression and protein production in follicles (oocytes and granulosa-cumulus cells) and corpus luteum (CL). In hypophysectomized ewes, GH and LH are necessary for normal CL development. IGF-1 mRNA in the follicles is expressed in theca interstitial cells (TIC) and granulosa cells (GC) with already higher levels in the TIC before follicle selection. In contrast, IGF-2 is mainly expressed in the TIC. The IGFR-1 mRNA is expressed in both the TIC and GC, with increasing levels in GC during the final development of dominant follicles. IGF-1 is a very potent stimulator of progesterone and oxytocin release in GC. IGFBP-1, -2, -3, -4, -5, and -6 have been isolated from follicular fluid or ovarian tissue. Studies indicate that IGFBP expression and production in the developing follicle is dependent on both cell type and follicle size and is regulated by IGF-1 and gonadotropins. The highest expression of IGF-1 and IGFR-1 mRNA was demonstrated during the early luteal phase. Distinct receptors for IGF-1 and IGF-2 were present in CL membrane preparations at all stages investigated. Intense immunostaining for IGF-1 was observed mainly in bovine large and small luteal cells and in a limited number of endothelial cells. In contrast, IGF-2 protein was localized in perivascular fibroblast and pericytes of the capillaries. With the use of a microdialysis system, we found that in vitro and in vivo IGF-1, IGF-2, and GH stimulated the release of progesterone in cultures of luteal cells or intact tissues. In conclusion, there is clear evidence for a central role of the IGFs, IGFBPs, and GH in follicular development and CL function.     

Effect of dietary concentrate content on feed intake,feed efficiency,and meat quality of Holstein steers fattened in a hot environment

The objective of this study was to investigate the effect of feeding high and low concentrate diets on feed intake and feed efficiency, the morphological characteristics of the rumen papillae, and meat quality of Holstein steers fattened under hot climate conditions in Oman. Ten male Holstein calves, of 5 months of age, were selected for the experiment. The animals were fed concentrate and Rhodes grass hay and were divided into two groups of high concentrate (HC, n = 5) and low concentrate diets (LC, n = 5), in which their feed intake, weight gain, and feed efficiency were evaluated across three growing periods. Feed intake and efficiency and average daily gain (ADG) of the HC group were significantly greater than for the LC group and were affected by the diet (p < .01) and the period (p < .001). Across the fattening periods, ADG declined in both groups, with ADG improved by 35% for steers on the HC diet compared to steers on the LC diet. Carcass meat quality was not affected significantly by the dietary concentrate level. In conclusion, our results can be used to make improvements in feed efficiency of Holstein steers under hot climate conditions.     

Effects of ronnel on growth, endocrine function and blood measurements in steers and rats

Four feeding trials were conducted to determine (1) how the organophosphate ronnel affects thyroid and adrenal circulating hormone levels and blood profile measurements of beef steers and (2) whether the effects of ronnel observed in beef steers could be produced in growing rats. In trial 1, four groups of eight steers each were given either no ronnel or 4 mg ronnel/kg body weight daily in diets fed at either limited (1.8% of body weight) or ad libitum intake. After 7 weeks, intake levels, but not ronnel treatments, were reversed during a 1-week transition period, and feeding was continued for another 7 weeks. In trial 2, four groups of six steers each were given 0, 44, 88 or 176 ppm ronnel premixed in diets fed ad libitum for 18 weeks. Actual ronnel intakes averaged 0, 1.01, 2.12 and 4.73 mg/kg body weight daily in trial 2. In trials 3 and 4 , growth and intake were measured in 64 and 32 growing Sprague-Dawley rats, respectively, fed levels of ronnel ranging from 0 to 100 ppm in the diet. In trial 3, blood plasma was analyzed at various times for triiodothyronine (T3), thyroxine (T4), cholesterol and total lipid content. In trial 1, the concentration of Plasma T4 was 1.31 times higher in steers fed ronnel that in control steers. In trial 2, plasma T4 was 1.30 times higher in steers fed 176 ppm ronnel, the level that improved growth, than in steers fed the lower ronnel levels. Circulating levels of T3, cortisol and aldosterone were similar for both control and ronnel-fed steers. Serum cholesterol concentrations were consistently higher in ronnel-fed steers. The data indicated that the growth-promoting effect of ronnel in steers may be associated with a shift in thyroid function. Effects of ronnel in steers were not observed in rats, demonstrating a species difference between steers and rats.     

Plasma growth hormone (GH) responses after administration of the peptidergic GH secretagogue KP102 into the oral cavity, rumen, abomasum and duodenum in adult goats

The study was performed to determine whether orally administered KP102 (also known as GHRP-2) stimulates GH release in adult goats, and how the orally administered KP102 passes through the digestive tract and stimulates GH release in ruminant animals. Five mg/kg body weight (BW) of KP102 dissolved in 9 ml of saline were administered into the oral cavity, rumen, omasum and duodenum of adult goats, and GH release after administration of KP102 was examined. The GH levels were significantly elevated at 20 min after administration of KP102 into the oral cavity, and plasma concentrations of GH remained significantly elevated until 60 min (P < 0.05). The GH levels after administration of KP102 into the abomasum were variable. However, the GH level tended to increase within 30 min after administration, and were significantly higher than those of controls after 120 to 150 min (P < 0.05). The GH levels after administration of KP102 into the duodenum were significantly elevated at 40 min after administration, and plasma concentrations of GH remained significantly elevated until 140 min (P < 0.05). The administration of KP102 into the rumen failed to stimulate GH release. The GH response curves (AUC) produced after administration of KP102 into the abomasum or duodenum were 2.2-fold greater than those for after administration into the oral cavity (P < 0.05). The oral administration of 5 mg/kg BW of KP102 in the powder state, not dissolved in 9 ml of saline, failed to stimulate GH release. These results suggested that orally administered KP102 dissolved in saline transiently stimulates GH release in adult goats, and this phenomenon might be due to small amounts of the peptides entering directly into the abomasum with liquid bypassing the rumen.     

Influence of prefast feed intake on recovery from feed and water deprivation by beef steers

A trial was conducted with 60 steers (257 kg) to determine the influence of prefast feed intake on recovery from feed and water deprivation. For 3 d, steers were fed a 35% roughage diet at 1 (LI) or 1.75% (MI) of body weight or ad libitum (AL). Steers were then deprived of feed and water for 24 h, limit-fed and watered for 24 h, deprived of feed and water for 48 h and then allowed ad libitum feed and water consumption for 2 wk. A fourth group of control steers was fed at 1.75% of body weight during the alimentation period and was not fasted. Realimentation feed intake was positively related to prefast feed intake, with the order of realimentation feed intake being AL greater than MI greater than LI (P less than .05). During deprivation, rumen volume declined (P less than .05) in AL-fed steers, but was not affected in LI and MI steers. Blood hemoglobin and serum urea-N increased during deprivation in all fasted groups. Prefast serum cholesterol levels were inversely related to prefast energy intake. During deprivation, rumen fluid total volatile fatty acid (VFA) concentrations and propionate and butyrate molar proportions declined (P less than .05) and acetate, isobutyrate and valerate + isovalerate molar proportions increased (P less than .05). Results of this study indicate that an increased prefast feed intake will provide a greater reserve of energy, water and electrolytes to the steer during deprivation and result in a shorter postfast adaptation period.     

Pregnancy-associated plasma protein-A and insulin-like growth factor binding protein mRNAs in granulosa cells of dominant and subordinate follicles of preovulatory cattle

To determine if (1) levels of pregnancy-associated plasma protein-A (PAPP-A) mRNA and insulin-like growth factor binding protein (IGFBP) (-2, -3, -4 and -5) mRNAs differ between the dominant and subordinate follicles during the follicular phase of an estrous cycle, and (2) these differences are associated with differences in follicular fluid (FFL) concentrations of steroids (estradiol, androstenedione, and progesterone), total and free IGF-I, or IGFBPs, estrous cycles of non-lactating Holstein dairy cows (n = 16) were synchronized with two injections of prostaglandin (PGF2 alpha) 11 days apart. Granulosa cells and FFL were collected either 24 h or 48 h after the second injection of PGF2 alpha. FFL from dominant follicles had lower concentrations of progesterone (P < 0.08) and higher concentrations of estradiol (P < 0.05), androstenedione (P < 0.0001), estradiol:progesterone ratio (P < 0.0001), free IGF-I (P < 0.0001), and calculated percentage free IGF-I (P < 0.01) than large subordinate follicles. Levels of IGFBP-2, -4, and -5 in FFL were 3.0- (P < 0.05), 2.4- (P < 0.06), and 3.4-fold (P < 0.05) greater, respectively, in subordinate than in dominant follicles. IGFBP-3, IGFBP-4 and PAPP-A mRNA expression and IGF-II concentration did not differ (P > 0.10) between dominant or subordinate follicles. Levels of IGFBP-2 and -5 mRNA were severalfold greater (P < 0.05) in subordinate than dominant follicles. IGFBP-5 mRNA in granulosa cells decreased (P < 0.05) 62% to 92%, between 24h and 48 h post-PGF2 alpha. We conclude that decreased levels of IGFBP-2 and -5 mRNA in granulosa cells may contribute to the decrease in FFL IGFBP-2 and -5 protein levels of preovulatory dominant follicles, and that changes in granulosa cell IGFBP-3 and -4 mRNA and PAPP-A mRNA levels do not occur during final preovulatory follicular development in cattle.     

Colostral and milk insulin-like growth factors and related substances: mammary gland and neonatal (intestinal and systemic) targets

The identification of hormones and regulatory factors in colostrum and milk has led to intensive investigations on their roles in the development and maintenance of the mammary and neonatal tissues. Insulin-like growth factors (IGFs) and IGF binding proteins (IGFBPs) in transgenic mice influence mammary biology gland towards the end of lactation. In the bovine, IGFBP-3 is the major IGFBP in mammary secretions. In addition to binding IGFs, IGFBP-3 also binds to lactoferrin (Lf). Secreted IGFBP-3 re-enters mammary epithelial cells and with the presence of a nuclear localization sequence, IGFBP-3 and Lf enter the nucleus. Nuclear IGFBP-3 affects apoptotic signaling through the retinoic-x-receptors, while Lf affects apoptotic events through unknown mechanisms. Such interactions likely influence mammary development and involution. Furthermore, ingested colostral bioactive factors can exert regulatory functions in neonates. Intestinal receptors for IGFs and insulin are modified by age and/or diet. Feeding IGF-I had no effect, but colostrum extracts had small intestinal effects (stimulation of proliferation and villus size), suggesting that several factors, rather than one single bioactive factor were responsible. Systemic changes of metabolic and endocrine profiles in neonates depend on composition, amounts, time and duration of feeding colostrum. Early postnatal colostrum intake is not only important for the provision and absorption of immunoglobulins. Thus, in neonatal calves the lack of colostrum intake during the first 24h after birth results in a low immunoglobulin G, beta-carotene and Vitamin A status that persists for weeks and plasma patterns of fatty acids, essential amino acids and the glutamine/glutamate ratios are affected. In calves oral administration of IGF-I had no and feeding of colostrum whey extracts had only minor effects on metabolic and endocrine traits. Thus, mammary secretions influence regulatory functions of mammary and neonatal tissues.     

Neutral detergent fiber concentration in corn silage influences dry matter intake, diet digestibility, and performance of Angus and Holstein steers

Twelve Angus (237 +/- 13 kg) and twelve Holstein (235 +/- 15 kg) steers were used to determine whether corn silage-based diets with different NDF levels influence DMI to a similar extent in Angus and Holstein steers and as body weight of the steers increase. Steers were randomly assigned to individual slatted-floor pens and used in a crossover design consisting of six 14-d periods. Experimental diets contained corn silage from a normal hybrid (low-fiber; LF) and its male-sterile counterpart (high-fiber; HF) and were alternated each period. The LF and HF diets contained 33.8 and 50.8% NDF, respectively. The HF diet decreased (P < 0.01) overall steer mean DMI 14.0% relative to LF, with mean differences increasing as steers increased in BW (P < 0.01). Feeding the HF diet also reduced ADG by an average of 13.8% relative to the LF diet (P < 0.01). Holstein steers consumed 14.4% more DM and gained 14.3% faster (P < 0.01) than Angus steers. There was a fiber level x breed-type interaction (P = 0.08) for efficiency of gain. Angus steers receiving the HF diet had greater efficiency of gain than Angus steers consuming the LF diet; however, Holstein steers consuming the LF diet had greater efficiency of gain than those receiving the HF diet. The HF treatment reduced total-tract digestibility of DM and GE by 4.6 and 4.5%, respectively (P < 0.01), and decreased DE intake by 20.5% (P < 0.01) but increased apparent totaltract digestibility of NDF and ADF (9.4 and 8.4%, respectively; P < 0.01). Holstein steers had similar digestibility of DM and GE (P > 0.10) but had greater DE intake (P < 0.01) compared to Angus steers. There were fiber level x breed-type interactions for total-tract digestibility of NDF and ADF (P < 0.06). The difference in DM digestibility was negatively associated with the difference in DMI (r2 = 0.23; P < 0.01) for LF minus HF within Angus steers, but not within Holstein steers (P = 0.42). Total-tract digestibility of NDF and ADF was 4.1 and 3.4% lower for the HF diet but was only 1.1 and 0.6% lower for the LF diet when fed to Holstein compared to Angus steers. Results from this trial demonstrate that high-NDF corn silage-based diets reduced intake of both Angus and Holstein steers, and this reduction in DMI continued as steers increased in BW from 235 to 330 kg. Breed differences were also noted for digestible energy intake as influenced by fiber level.     

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.     

Cloning and expression of equine insulin-like growth factor binding proteins in normal equine tendon

OBJECTIVES: To define a portion of the nucleotide sequences of each of the 6 insulin-like growth factor (IGF) binding proteins (IGFBPs) in horses and describe patterns of messenger RNA (mRNA) and protein expression for IGFBPs in normal equine tendons. ANIMALS: 7 horses. PROCEDURE: Total RNA was extracted from the tensile region of normal superficial digital flexor tendons and reverse transcribed into complimentary DNA (cDNA). The cDNA was amplified via PCR, and products representing portions of each IGFBP were cloned and sequenced. Nucleotide sequences were used to deduce the amino acid sequences, and both nucleotide and predicted amino acid sequences were compared with those published for bovine, human, mouse, and ovine IGFBPs. Gene expression was quantitated by real-time PCR assay, and protein expression was evaluated by western ligand blot (WLB). RESULTS: Clones ranged in size from 262 to 522 bp and had high degrees of sequence homology with other mammalian species. Sequence homology was highest between bovine and equine IGFBPs (86% to 95%) and amongst the IGFBP-5 sequences from the various species (92% to 95%). Message for IGFBP-2 to -6, but not IGFBP-1, was expressed in normal tendon. Protein expression for IGFBP-2, -3, and -4 was detected byWLB in normal tendon and markedly increased in damaged tendons. CONCLUSIONS AND CLINICAL RELEVANCE: Results provide basic information and tools needed for further characterization of the role of the IGF system in tendon healing and may lead to the ability to potentiate the response of healing tendon to exogenous IGF-I via concurrent manipulation of IGFBPs.