Ultrasound during mid-gestation: Agreement with physical foetal and placental measurements and use in predicting gestational age in sheep.

To determine the effects of poor maternal nutrition and litter size on foetal growth during mid-gestation, pregnant ewes (n = 82) were fed 100%, 60% or 140% of NRC TDN beginning at day 30.2 ± 0.2 of gestation. Transabdominal ultrasound was performed weekly between day 46.0 ± 0.4 and 86.0 ± 0.7 to monitor foetal heart width (HW), umbilical diameter (UMB), rib width (RW) and placentome outer (OD) and inner diameter (ID). Data were analysed with repeated-measures using the mixed procedure for effects of maternal diet, litter size and gestation, and equations predictive of gestational age were generated using the regression procedure. To determine the agreement of ultrasound measurement and actual size, ewes (n = 20-21) were euthanized at day 45 or 90 to obtain corresponding postmortem measurements for Bland-Altman analysis. The HW, UMB and placentome OD and ID increased with gestation (p < .0001) but were unaffected by maternal diet or litter size (p ≥ .12). Ultrasound underestimated postmortem measurements of HW (14.8%), UMB (7.3%), placentome OD (4.5%) and ID (37.3%) at day 90 of gestation. Ultrasound underestimated RW at day 45 (7.7%) but overestimated RW (23.8%) at day 90, indicating inconsistent bias when reporting RW by ultrasound. Combining the HW, UMB, RW and placentome OD generated the strongest equation predictive of gestational age (R2  = .91). These findings indicate that during mid-gestation, maternal diet or litter size did not affect HW, UMB or placentome diameters and these factors can be used to estimate gestational age.

Effect of daily lithium chloride administration on bone mass and strength in growing broiler chickens.

The objective was to determine the effects of oral lithium chloride supplementation on bone strength and mass in broiler chickens. Ninety-six broilers were assigned to 1 of 2 treatment groups (lithium chloride or control; n=48/treatment). Beginning at 1 or 3 wk of age, chickens were administered lithium chloride (20 mg/kg body weight) or water daily by oral gavage. At 6 wk of age, chickens were euthanized and bone and muscle samples were collected. A 24 h lithium chloride (20 mg/kg body weight) challenge determined that serum lithium chloride increased within 2 h and cleared the system within 24 h, demonstrating the effective delivery of lithium chloride. Treatment did not influence body weight (P≥0.20) or feed intake (P≥0.81), demonstrating that lithium chloride did not negatively affect broiler growth. To determine bone strength, 3-point bending was performed on the femora and tibiae obtained from control and lithium chloride-treated birds in the 1 wk group. Lithium chloride-treated birds had a 22% reduction in stiffness compared with control in the femora (P=0.02) without a corresponding reduction in elastic modulus. No differences were observed in yield or ultimate load and in the corresponding calculations of stresses (P≥0.26). The toughness of tibiae was not altered in lithium chloride compared with control (P=0.11). Bone length and micro-CT imaging were performed on the tibiae of control and lithium chloride groups. No differences (P≥0.52) in bone length, cortical or trabecular bone volume, trabecular thickness, number, or spacing were observed. Lithium chloride treatment did not affect pectoralis muscle color or lipid oxidation (P>0.05). In conclusion, lithium chloride treatment in broilers did not negatively affect growth or meat quality. A reduction in bone stiffness of the femur with lithium chloride treatment was observed, however unlike the mouse model, the dosages of lithium chloride used in the current study did not result in anabolic effects on broiler long bones.

Short communication: Expression of T-box 2 and 3 in the bovine mammary gland.

To increase our understanding of the mechanisms by which growth hormone (GH) and insulin-like growth factor (IGF)-I influence bovine mammary gland development, the potential roles of T-box2 (TBX2) and T-box3 (TBX3) were investigated. Although no information regarding expression of either transcription factor in the bovine mammary gland exists, it is known that TBX3 and its closely related family member, TBX2, are required for mammary gland development in humans and mice. Additionally, TBX3 mutations in humans and mice lead to ulnar mammary syndrome. Evidence is present in bone that TBX3 is required for proliferation and its expression is regulated by GH, an important regulator of mammary gland development and milk production. We hypothesized that TBX2 and TBX3 are expressed in the bovine mammary gland and that GH, IGF-I, or both increase TBX2 and TBX3 expression in bovine mammary epithelial cells (MEC). Bovine mammary gland tissue, MAC-T cells, primary MEC, and fibroblasts were obtained and TBX2 and TBX3 expression was determined by real-time reverse transcription PCR. In addition, TBX2 and TBX3 expression was examined in cells treated with 100 or 500 ng/mL of GH or 100 or 200 ng/mL of IGF-I for 24 or 48 h. Both TBX2 and TBX3 were expressed in bovine mammary tissue. Surprisingly, expression of TBX2 was only detected in mammary fibroblast cells, whereas TBX3 was expressed in all 3 cell types. Growth hormone did not alter TBX3 expression in MAC-T cells or MEC. However, IGF-I increased TBX3 expression in MAC-T, but not in primary MEC. We did not observe a change in TBX2 or TBX3 expression in fibroblasts treated with GH and IGF. Therefore, we concluded that (1) TBX2 and TBX3 are expressed in bovine mammary gland, (2) their expression is cell-type specific, and (3) IGF-I stimulates TBX3 expression in MAC-T cells.

Effects of poor maternal diet during gestation are detected in F2 offspring.

Poor maternal nutrition of F0 ewes impairs F1 offspring growth, with minimal differences in glucose tolerance or select metabolic circulating factors, and independent of differences in residual feed intake (RFI). To determine if poor maternal nutrition in F0 ewes alters F2 offspring growth, circulating leptin, feed efficiency, or glucose tolerance, F0 ewes (n = 46) pregnant with twins were fed 100% (control), 60% (restricted), or 140% (over) of National Research Council requirements from days 30 ±â€…0.02 of gestation until parturition. At 16 to 19 mo of age, female F1 (n = 36) offspring were bred to generate F2 offspring [CON-F2 (n = 12 ewes; 6 rams), RES-F2 (n = 7 ewes; 13 rams), or OVER-F2 (n = 13 ewes; 9 rams) corresponding to diets of the granddam (F0)]. Lamb body weights (BW) and blood samples were collected weekly from days 0 to 28 and every 14 d until day 252 of age. Circulating leptin was measured in serum at days 0, 7, 14, 56, 210, and 252. An intravenous glucose tolerance test was performed at days 133 ±â€…0.28. At days 167 ±â€…0.33, individual daily intake was recorded over a 77-d feeding period to determine RFI. Rams were euthanized at days 285 ±â€…0.93, and body morphometrics, loin eye area (LEA), back fat thickness, and organ weights were collected and bone mineral density (BMD) and length were determined in the right hind leg. During gestation, OVER-F1 ewes tended to be 8.6% smaller than CON-F1 ewes (P ≤ 0.06). F2 offspring were of similar BW from birth to day 70 (P ≥ 0.20). However, from days 84 to 252, RES-F2 offspring tended to be 7.3% smaller than CON-F2 (P ≤ 0.10). Granddam diet did not influence F2 ram body morphometrics, organ or muscle weights, LEA, adipose deposition, or leg BMD (P ≥ 0.84). RES-F2 (-0.20) and CON-F2 (-0.45) rams tended to be more feed efficient than CON-F2 ewes (0.31; P ≤ 0.08). No effects of granddam diet were observed on glucose or insulin average or baseline concentrations, area under the curve, first-phase response, or ratio (P ≥ 0.52). However, CON-F2 rams (297 mg/dL ±â€…16.5) had a greater glucose peak compared with RES-F2 rams (239 mg/dL ±â€…11.2; P = 0.05). Peak insulin concentrations were not influenced by granddam diet (P = 0.75). At d 56, RES-F2 and OVER-F2 offspring had 53.5% and 61.8% less leptin compared with CON-F2 offspring, respectively (P ≤ 0.02). These data indicate that poor maternal nutrition impacts offspring growth into the second generation with minimal impacts on offspring RFI, glucose tolerance, and circulating leptin.

Survey of the use of beef semen in dairy herds in Pennsylvania and nearby states.

Because dairies across the United States have rapidly adopted breeding to beef breed sires, the use of beef semen has increased dramatically in recent years. The objective of this survey was to gather information about the use of beef semen by dairy producers in the Northeast United States to generate beef × dairy cattle for beef markets. The survey was conducted using the services of the Center for Survey Research at the Pennsylvania State University-Harrisburg campus. Respondents had two options for returning their responses: 1) mail the paper survey to CSR in the postage-paid business-reply envelope included in the mailing, or 2) complete the survey online via an open-access web survey link. A total of 669 surveys were received and a final number of 617 surveys were included in the responses based on completeness and validity of the responses. Because of the broad electronic distribution, a true response rate cannot be calculated. Of these, 463 (75.0%) were completed via returned paper survey, and 154 (25.0%) were completed via web, between November 9, 2021 and February 16, 2022. Of the 617 respondents, 539 were from Pennsylvania. Due to the large variations in returned survey copies by state, results are reported without state separation. Across all respondents, 69.7% reported milking 100 or fewer cows and over 90% of collected responses reported Holsteins as the predominant dairy breed in the Northeast. Only 18.8% of the respondents did not currently, nor plan to, breed with beef semen. Deciding which beef bulls to use on Northeast dairy farms was primarily based on the recommendation of the semen sales representative (54.5%) and the price of the semen purchased (42.3%). In addition, 89.7% of respondents cited using Angus genetics in their beef bull selections. However, there was no difference in reported profitability of crossbreeding between respondents who indicated using other beef breeds vs. those who indicated just using Angus (P ≥ 0.19). In conclusion, using beef sires on dairy females, regardless of the breed of beef sire, adds value to the resulting progeny from dairy farms in the Northeast.

Effects of poor maternal nutrition during gestation on ewe and offspring plasma concentrations of leptin and ghrelin.

Poor maternal nutrition during gestation can negatively affect offspring growth, development, and health. Leptin and ghrelin, key hormones in energy homeostasis and appetite control, may mediate these changes. We hypothesized that restricted- and over-feeding during gestation would alter plasma concentrations of leptin and ghrelin in ewes and offspring. Pregnant ewes (n = 37) were fed 1 of 3 diets starting on d 30 ± 0.02 of gestation until necropsy at d 135 of gestation or parturition: restricted- [RES; 60% National Research Council (NRC) requirements for total digestible nutrients, n = 13], control- (CON; 100% NRC, n = 11), or over-fed (OVER; 140% NRC, n = 13). Blood samples were collected from pregnant ewes at days 20, 30, 44, 72, 100, 128, and 142 of gestation. Offspring blood samples were collected within 24 h after birth (n = 21 CON, 25 RES, 23 OVER). Plasma leptin and ghrelin concentrations were determined by RIA. Ewe data were analyzed using the MIXED procedure in SAS with ewe as the repeated subject. Offspring data were analyzed using the MIXED procedure. Correlations between BW and leptin and ghrelin concentrations were identified using PROC CORR. At d 100, RES (5.39 ± 2.58 ng/mL) had decreased leptin concentrations compared with OVER (14.97 ± 2.48 ng/mL; P = 0.008) and at d 128, RES (6.39 ± 2.50 ng/mL) also had decreased leptin concentrations compared with OVER (13.61 ± 2.47 ng/mL; P = 0.04). At d 142, RES (0.26 ± 0.04 ng/mL) had increased ghrelin concentrations compared with CON (0.15 ± 0.04 ng/mL; P = 0.04). Leptin and ghrelin concentrations were also altered between days of gestation within a dietary treatment. In CON ewes, plasma concentrations of leptin were increased at d 30 (19.28 ± 7.43 ng/mL) compared with d 44 (5.20 ± 3.10 ng/mL; P = 0.03), and the plasma concentrations of ghrelin at d 128 (0.20 ± 0.03 ng/mL) were increased compared with d 30 (0.16 ± 0.03 ng/mL; P = 0.01) and d 100 (0.17 ± 0.03 ng/mL; P = 0.04). Maternal diet did not alter plasma ghrelin or leptin concentrations in the offspring (P > 0.50). There were no strong, significant correlations between ewe BW and leptin (r < 0.33; P > 0.06) or ghrelin (r > -0.47; P > 0.001) concentrations or lamb BW and leptin or ghrelin concentrations (r > -0.32, P > 0.06). Maternal alterations in circulating leptin and ghrelin may program changes in energy balance that could result in increased adiposity in adult offspring. Alterations in energy homeostasis may be a mechanism behind the long-lasting changes in growth, body composition, development, and metabolism in the offspring of poorly nourished ewes.

T-box 3 negatively regulates osteoblast differentiation by inhibiting expression of osterix and runx2.

T-box (Tbx)3, a known transcriptional repressor, is a member of a family of transcription factors, which contain a highly homologous DNA binding domain known as the Tbx domain. Based on the knowledge that mutation of the Tbx3 gene results in limb malformation, Tbx3 regulates osteoblast proliferation and its expression increases during osteoblast differentiation, we predicted that Tbx3 is an important regulator of osteoblast cell functions. In this study, we evaluated the consequence of transgenic overexpression of Tbx3 on osteoblast differentiation. Retroviral overexpression increased Tbx3 expression >100-fold at the mRNA and protein level. Overexpression of Tbx3 blocked mineralized nodule formation (28 +/- 8 vs. 7 +/- 1%) in MC3T3-E1 cells. In support of these data, alkaline phosphatase (ALP) activity was reduced 33-70% (P < 0.05) in both MC3T3-E1 cells and primary calvaria osteoblasts overexpressing Tbx3. In contrast, Tbx3 overexpression did not alter ALP activity in bone marrow stromal cells. Tbx3 overexpression blocked the increase in expression of key osteoblast marker genes, ALP, bone sialoprotein, and osteocalcin that occurs during normal osteoblast differentiation, but had little or no effect on expression of proliferation genes p53 and Myc. In addition, Tbx3 overexpression abolished increased osterix and runx2 expression observed during normal osteoblast differentiation, but the change in Msx1 and Msx2 expression over time was similar between control and Tbx3 overexpressing cells. Interestingly, osterix and runx2, but not Msx1 and Msx2, contain Tbx binding site in the regulatory region. Based on these data and our previous findings, we conclude that Tbx3 promotes proliferation and suppresses differentiation of osteoblasts and may be involved in regulating expression of key transcription factors involved in osteoblast differentiation.

Complex genetic regulation of bone mineral density and insulin-like growth factor-I in C57BL/6J-Chr #A/J/NaJ chromosome substitution strains.

Low bone mineral density (BMD) is a phenotype associated with osteoporosis and increased risk of fracture. Since 60-80% of variation in BMD is associated with genetic factors, we used the novel approach of chromosome substitution strains (CSS) to identify chromosomes that harbor genes that regulate BMD. Specifically, we evaluated 24 wk old C57BL/6J-Chr #(A/J)/NaJ CSS (n = 7 to 18) in which each chromosome in the host C57BL/6J strain is replaced by a corresponding chromosome from the donor A/J strain (19 autosomes, X, Y). We determined several A/J chromosomes contribute to body weight (BW), percent body fat (BF), whole body areal BMD (aBMD), and serum insulin-like growth factor (IGF)-I in both a positive and negative manner when compared with C57BL/6J. Specifically, C57BL/6J-Chr 5(A/J)/NaJ (B.A-5) (males) and B.A-13 (females) contributed to increased BW, whereas B.A-3, 4, 8, 9, 12, and 18 (males) and B.A-3, 4, and 11 (females) contributed to reduced BW. B.A-5 (males) and B.A-13 (females) contributed to increased BF, whereas B.A-12 (males) and B.A-3, 4, 10, and 11 (females) contributed to reduced BF. B.A-14 (females) contributed to increased aBMD and B.A-1, 2, 6, 9, 10, and 18 (males) and B.A-8, 9, and 10 (females) contributed to reduced aBMD. To determine if similar chromosomes regulate aBMD and IGF-I, we determined serum concentrations of IGF-I. B.A-14 and Y (males) and B.A-6 (females) contributed to increased IGF-I and male B.A-3 and female B.A-8 contributed to reduced IGF-I. Overall, we identified several sex-dependent and -independent chromosomes that regulate aBMD and IGF-I in adult mice.

PHYSIOLOGY AND ENDOCRINOLOGY SYMPOSIUM:The effects of poor maternal nutrition during gestation on offspring postnatal growth and metabolism.

Poor maternal nutrition during gestation has been linked to poor growth and development, metabolic dysfunction, impaired health, and reduced productivity of offspring in many species. Poor maternal nutrition can be defined as an excess or restriction of overall nutrients or specific macro- or micronutrients in the diet of the mother during gestation. Interestingly, there are several reports that both restricted- and over-feeding during gestation negatively affect offspring postnatal growth with reduced muscle and bone deposition, increased adipose accumulation, and metabolic dysregulation through reduced leptin and insulin sensitivity. Our laboratory and others have used experimental models of restricted- and over-feeding during gestation to evaluate effects on early postnatal growth of offspring. Restricted- and over-feeding during gestation alters body size, circulating growth factors, and metabolic hormones in offspring postnatally. Both restricted- and over-feeding alter muscle growth, increase lipid content in the muscle, and cause changes in expression of myogenic factors. Although the negative effects of poor maternal nutrition on offspring growth have been well characterized in recent years, the mechanisms contributing to these changes are not well established. Our laboratory has focused on elucidating these mechanisms by evaluating changes in gene and protein expression, and stem cell function. Through RNA-Seq analysis, we observed changes in expression of genes involved in protein synthesis, metabolism, cell function, and signal transduction in muscle tissue. We recently reported that satellite cells, muscle stem cells, have altered expression of myogenic factors in offspring from restricted-fed mothers. Bone marrow derived mesenchymal stem cells, multipotent cells that contribute to development and maintenance of several tissues including bone, muscle, and adipose, have a 50% reduction in cell proliferation and altered metabolism in offspring from both restricted- and over-fed mothers. These findings indicate that poor maternal nutrition may alter offspring postnatal growth by programming stem cell populations. In conclusion, poor maternal nutrition during gestation negatively affects offspring postnatal growth, potentially through impaired stem and satellite cell function. Therefore, determining the mechanisms that contribute to fetal programming is critical to identifying effective management interventions for these offspring and improving efficiency of production.

Fetal and organ development at gestational days 45, 90, 135 and at birth of lambs exposed to under- or over-nutrition during gestation,.

To determine the effects of poor maternal nutrition on offspring body and organ growth during gestation, pregnant Western White-faced ewes (n = 82) were randomly assigned into a 3 × 4 factorial treatment structure at d 30.2 ± 0.2 of gestation (n = 5 to 7 ewes per treatment). Ewes were individually fed 100% (control), 60% (restricted) or 140% (over) of NRC requirements for TDN. Ewes were euthanized at d 45, 90 or 135 of gestation or underwent parturition (birth) and tissues were collected from the offspring (n = 10 to 15 offspring per treatment). Offspring from control, restricted and overfed ewes are referred to as CON, RES and OVER, respectively. Ewe data were analyzed as a completely randomized design and offspring data were analyzed as a split-plot design using PROC MIXED. Ewe BW did not differ at d 30 (P ≥ 0.43), however restricted ewes weighed less than overfed and overfed were heavier than controls at d 45, and restricted weighed less and overfed were heavier than controls at d 90 and 135 and birth (P ≤ 0.05). Ewe BCS was similar at d 30, 45 and 90 (P ≤ 0.07), however restricted ewes scored lower than control at d 135 and birth (P ≤ 0.05) and over ewes scored higher than control at d 135 (P ≤ 0.05) but not at birth (P = 0.06). A maternal diet by day of gestation interaction indicated that at birth the body weight (BW) of RES offspring was less than CON and OVER (P ≤ 0.04) and heart girth of RES was smaller than CON and OVER (P ≤ 0.004). There was no interaction of maternal diet and day of gestation on crown-rump, fetal, or nose occipital length, or orbit or umbilical diam. (P ≥ 0.31). A main effect of maternal diet indicated that the RES crown-rump length was shorter than CON and OVER (P ≤ 0.05). An interaction was observed for liver, kidney and renal fat (P ≤ 0.02). At d 45 the liver of RES offspring was larger than CON and OVER (P ≤ 0.002), but no differences observed at d 90, 135 or birth (P ≥ 0.07). At d 45, the kidneys of OVER offspring were larger than CON and RES (P ≤ 0.04), but no differences observed at d 90, 135 or birth (P ≥ 0.60). At d 135, OVER had more perirenal fat than CON and RES (P ≤ 0.03), and at birth RES had more perirenal fat than CON and OVER (P ≤ 0.04). There was no interaction observed for offspring heart weight, length or width, kidney length, adrenal gland weight, loin eye area or rib width (P ≥ 0.09). In conclusion, poor maternal nutrition differentially alters offspring body size and organ growth depending on the stage of gestation.

Regulation of insulin-like growth factor binding protein-5, four and a half lim-2, and a disintegrin and metalloprotease-9 expression in osteoblasts.

The roles of insulin-like growth factors (IGFs) in regulating growth and their modulation by six IGF binding proteins (IGFBP) are well established. IGFBP-5, the most abundant IGFBP stored in bone, is an important regulator of bone formation via IGF-dependent and -independent mechanisms. Two new proteins, four and a half lim (FHL)-2, a transcription modulator that interacts with IGFBP-5, and a disintegrin and metalloprotease (ADAM)-9, an IGFBP-5 protease, have been identified as potential regulators of IGFBP-5 action in bone. We tested the hypothesis that agents which modulate bone formation by regulating IGFBP-5 expression would also regulate FHL-2 and ADAM-9 expression in a coordinated manner. We evaluated the expression of IGFBP-5, FHL-2, and ADAM-9 by real-time reverse transcriptase (RT)-PCR during differentiation of mouse bone marrow stromal cells into osteoblasts and in response to treatment with bone formation modulators in the LSaOS human osteosarcoma cell line. IGFBP-5 and FHL-2 increased 4.3- and 3.0-fold (P < or = 0.01), respectively, during osteoblast differentiation. Dexamethasone (Dex), an inhibitor of bone formation, decreased IGFBP-5 and FHL-2 and increased ADAM-9 in LSaOS cells (P < or = 0.05). Bone morphogenic protein (BMP)-7, a stimulator of bone formation, increased IGFBP-5 and decreased ADAM-9 (P<0.01). To determine if BMP-7 would eliminate Dex inhibition of IGFBP-5, cells were treated with Dex+BMP-7. The BMP-7-induced increase in IGFBP-5 was reduced, but not eliminated, in the presence of Dex (P < or = 0.01), indicating that BMP-7 and Dex may regulate IGFBP-5 via different mechanisms. Transforming growth factor (TGF)-beta, a stimulator of bone formation, increased IGFBP-5 and FHL-2 expression (P < or = 0.01). IGF-I and TNF-alpha decreased expression of ADAM-9 (P<0.05). In conclusion, our findings are consistent with the hypothesis that FHL-2 and ADAM-9 are important modulators of IGFBP-5 actions and are, in part, regulated in a coordinated manner in bone.

Restricted maternal nutrition alters myogenic regulatory factor expression in satellite cells of ovine offspring.

Poor maternal nutrition inhibits muscle development and postnatal muscle growth. Satellite cells are myogenic precursor cells that contribute to postnatal muscle growth, and their activity can be evaluated by the expression of several transcription factors. Paired-box (Pax)7 is expressed in quiescent and active satellite cells. MyoD is expressed in activated and proliferating satellite cells and myogenin is expressed in terminally differentiating cells. Disruption in the expression pattern or timing of expression of myogenic regulatory factors negatively affects muscle development and growth. We hypothesized that poor maternal nutrition during gestation would alter the in vitro temporal expression of MyoD and myogenin in satellite cells from offspring at birth and 3 months of age. Ewes were fed 100% or 60% of NRC requirements from day 31±1.3 of gestation. Lambs from control-fed (CON) or restricted-fed (RES) ewes were euthanized within 24 h of birth (birth; n=5) or were fed a control diet until 3 months of age (n=5). Satellite cells isolated from the semitendinosus muscle were used for gene expression analysis or cultured for 24, 48 or 72 h and immunostained for Pax7, MyoD or myogenin. Fusion index was calculated from a subset of cells allowed to differentiate. Compared with CON, temporal expression of MyoD and myogenin was altered in cultured satellite cells isolated from RES lambs at birth. The percent of cells expressing MyoD was greater in RES than CON (P=0.03) after 24 h in culture. After 48 h of culture, there was a greater percent of cells expressing myogenin in RES compared with CON (P0.05). In satellite cells from RES lambs at 3 months of age, the percent of cells expressing MyoD and myogenin were greater than CON after 72 h in culture (P<0.05). Fusion index was reduced in RES lambs at 3 months of age compared with CON (P<0.001). Restricted nutrition during gestation alters the temporal expression of myogenic regulatory factors in satellite cells of the offspring, which may reduce the pool of myoblasts, decrease myoblast fusion and contribute to the poor postnatal muscle growth previously observed in these animals.

Poor maternal nutrition during gestation alters the expression of genes involved in muscle development and metabolism in lambs.

Poor maternal nutrition during gestation can result in reduced muscle mass and increased adiposity of the muscle tissue in the offspring. This can have long-lasting consequences on offspring health and productivity. However, the mechanisms by which poor maternal nutrition affects postnatal muscle development are poorly understood. We hypothesized that poor maternal nutrition during gestation would alter expression of key pathways and genes involved in growth, development, and maintenance of the muscle of lambs. For this study, beginning at d 31 ± 1.3 of gestation, ewes were fed 100 (control), 60 (restricted), or 140% (overfed) of the NRC requirements. Within 24 h of birth, lambs were necropsied and semitendinosus muscle tissue was collected for gene expression analysis. Using RNA sequencing (RNA-seq) across dietary treatment groups, 35 and 10 differentially expressed genes were identified using the and reference annotations, respectively. Maternal overfeeding caused changes in the expression of genes involved in regulating muscle protein synthesis and growth as well as metabolism. Alternately, maternal nutrient restriction affected genes that are involved in muscle cell proliferation and signal transduction. That is, despite a similar phenotype, the genes identified differed between offspring born to restricted- or overfed, ewes indicating that the mechanism for the phenotypic changes in muscle are due to different mechanisms.

The effects of poor maternal nutrition during gestation on postnatal growth and development of lambs.

Poor maternal nutrition can affect the growth and development of offspring, which may lead to negative consequences in adult life. We hypothesized that lambs born to poorly nourished ewes would have reduced growth rate and increased fat deposition, with corresponding changes in the somatotropic axis, and leptin, insulin and glucose concentrations. Ewes ( = 36; 12/treatment) were assigned 1 of 3 diets; 100% (CON), 60% (RES), or 140% (OVER) of NRC requirements for TDN at d 31 of gestation until parturition. One lamb per ewe ( = 35; 11 to 12 per treatment) was used; 18 lambs were euthanized at d 1, and 17 were fed the same diet for 3 mo and then euthanized. Lamb crown rump length (CRL), heart girth, BW, and BCS were measured, and blood samples were collected at d 1 and then at weekly intervals until euthanasia. Averaged from d 1 until 3 mo, lambs from OVER ewes were larger compared with lambs born to CON ewes (BW [16.97 vs. 15.44 kg ± 0.60; = 0.09], ADG [0.23 vs. 0.21 ± 0.01 kg/d; = 0.01], and CRL [68.9 vs. 66.1 ± 0.80 cm; = 0.02]). On a BW basis, heart weight from lambs from RES (0.18 kg ± 0.03; = 0.03) ewes was greater than that of CON lambs (0.15 kg ± 0.03). Backfat thickness was reduced in RES lambs (0.11 ± 0.06; ≤ 0.04) compared with CON (0.20 ± 0.06) and OVER (0.26 ± 0.06) lambs. Concentrations of IGF-I at 3 mo and IGFBP-3 from weaning (d 56 of age) to 3 mo of age tended to be greater ( ≤ 0.06) in OVER lambs (334 ± 66 ng/mL and 175 ± 11 arbitrary units [AU], respectively) than CON lambs (149 ± 66 ng/mL and 140 ± 11 AU, respectively). At 3 mo, leptin was greater in OVER lambs compared with RES lambs (1.24 vs. 0.78 ± 0.13 ng/mL; < 0.05). Over time, average insulin concentrations were greater in OVER and RES lambs than CON lambs (0.49 and 0.49 vs. 0.33 ± 0.05 ng/mL; ≤ 0.02). However, concentrations of GH, IGFBP-2, glucose, triglycerides, and total cholesterol were not different ( > 0.10) between treatment groups. During in vivo glucose tolerance test, baseline insulin concentrations were 68% and 85% greater ( 0.01), respectively, in RES and OVER lambs compared with CON lambs. Similarly, the glucose:insulin ratio was greater in RES and OVER lambs compared with CON lambs ( 0.01). Thus, in this experiment, poor maternal nutrition during gestation influenced body size, organ growth, fat accumulation, and concentrations of IGF-I, IGFBP-3, leptin, and insulin of offspring during the first 3 mo of age.

HORSE SPECIES SYMPOSIUM: Use of mesenchymal stem cells in fracture repair in horses.

Equine bone fractures are often catastrophic, potentially fatal, and costly to repair. Traditional methods of healing fractures have limited success, long recovery periods, and a high rate of reinjury. Current research in the equine industry has demonstrated that stem cell therapy is a promising novel therapy to improve fracture healing and reduce the incidence of reinjury; however, reports of success in horses have been variable and limited. Stem cells can be derived from embryonic, fetal, and adult tissue. Based on the ease of collection, opportunity for autologous cells, and proven success in other models, adipose- or bone marrow-derived mesenchymal stem cells (MSC) are often used in equine therapies. Methods for isolation, proliferation, and differentiation of MSC are well established in rodent and human models but are not well characterized in horses. There is recent evidence that equine bone marrow MSC are able to proliferate in culture for several passages in the presence of autologous and fetal bovine serum, which is important for expansion of cells. Mesenchymal stem cells have the capacity to differentiate into osteoblasts, the bone forming cells, and this complex process is regulated by a number of transcription factors including runt-related transcription factor 2 (Runx2) and osterix (Osx). However, it has not been well established if equine MSC are regulated in a similar manner. The data presented in this review support the view that equine bone marrow MSC are regulated by the same transcription factors that control the differentiation of rodent and human MSC into osteoblasts. Although stem cell therapy is promising in equine bone repair, additional research is needed to identify optimal methods for reintroduction and potential manipulations to improve their ability to form new bone.

The ontogeny of the somatotropic axis in Hereford calves from birth to one year of age and its response to administration of exogenous bovine somatotropin.

Administration of exogenous bovine ST (bST) increases growth rate, feed efficiency, and carcass quality in beef cattle. The magnitude of response to bST in beef cattle is variable and related to the age of the animal. Our objective was to determine the response of the somatotropic axis, in particular IGF-I, IGFBP-2, and IGFBP-3, to bST treatment from birth to 1 yr of age. Blood samples were collected before and after a single injection of bST (500 mg) every 50 d from birth to 1 yr of age in male and female Hereford calves. Body weights and serum concentrations of ST, IGF-I, IGFBP-2, and IGFBP-3 were determined. At birth, serum concentrations of ST, IGF-I, and IGFBP-3 increased (P < 0.05) following bST treatment. From 50 to 350 d of age, average concentrations of ST and IGF-I were greater (P < 0.05) in males, whereas IGFBP-2 concentrations were greater (P < 0.05) in females. No gender differences in IGFBP-3 concentrations were observed. Following bST treatment, IGF-I increased (P < 0.05) from 50 to 350 d of age, IGFBP-2 decreased (P < 0.05) from 50 to 200 d of age, and IGFBP-3 increased (P < 0.05) at 250 d of age. At 250 d of age, baseline concentrations of IGFBP-2 decreased (P < 0.05). Due to the positive response of IGFBP-3 and decreased baseline IGFBP-2 at 250 d of age, we conclude that this is an age at which the somatotropic axis is most responsive to exogenous bST, and it therefore may be an appropriate age to begin bST treatment in beef calves to realize the positive influence of bST on BW gain, feed efficiency, and carcass composition.

The ontogeny of the somatotropic axis in male and female Hereford calves from birth to one year of age.

Insulin-like growth factor-binding proteins-2 and -3 may play a role in age-dependent growth response to bovine ST (bST) treatment in cattle; however, samples have been collected at infrequent intervals and at limited time points. Therefore, the objective of this experiment was to examine the ontogeny of components of the somatotropic axis in Hereford calves from birth to 1 yr of age at weekly intervals to determine whether there is a certain age or time frame when the somatotropic axis may change and/or potentially become more responsive to exogenous bST administration. Blood samples and body weight measurements were collected from eight male and eight female Hereford calves once per week from birth to 1 yr of age. Serum concentrations of ST, IGF-I, IGFBP-2, and IGFBP-3 were determined. Males began to grow faster than females at approximately 16 wk of age (P < 0.05). Average concentrations of ST, IGF-I, and IGFBP-3 were greater in males than females (P < 0.01). Average concentrations of IGFBP-2 were greater in females than in males (P = 0.05). Concentrations of ST decrease with age (P < 0.01); however, the decrease occurred earlier in female calves. Concentrations of IGF-I and IGFBP-3 increased in males and females (P < 0.01), and concentrations of IGF-I began to plateau at approximately the same time as growth rate differences were observed (16 wk of age). Following an initial increase (birth to approximately 16 wk of age), concentrations of IGFBP-3 remained constant until approximately 43 wk of age. Concentrations of IGFBP-2 increased to approximately 10 wk of age (P < 0.05), followed by a decrease, and then, similar to IGFBP-3, remained constant until 43 wk of age. Correlations between average daily gain, ST, IGF-I, IGFBP-2, and IGFBP-3 were determined. Average daily gain was negatively (P < 0.01) correlated with ST and positively (P < 0.1) correlated with IGF-I. In females, ST was negatively (P < 0.01) correlated with IGF-I. Concentrations of ST were positively correlated (P < 0.01) with IGFBP-2 and IGFBP-3. Concentrations of IGFBP-2 were negatively correlated (P < 0.01) with IGF-I and positively correlated (P < 0.01) with IGFBP-3. In conclusion, serum concentrations of ST, IGF-I, IGFBP-2, and IGFBP-3 differed between male and fe-male calves. In addition, changes in components of the somatotropic axis occurred around the same time as males began to grow faster than females.

The influence of level of feeding on growth and serum insulin-like growth factor I and insulin-like growth factor-binding proteins in growing beef cattle supplemented with somatotropin.

The objective of this study was to determine the effects of level of feeding on growth, feed efficiency (gain:feed; G:F), body composition (BC), and serum concentrations of somatotropin (ST), IGF-I, and IGF-binding proteins (BP) in growing beef cattle supplemented with bovine (b) ST. In each of two consecutive years, 40 growing beef cattle were blocked by weight (average BW: yr 1 = 316 kg, yr 2 = 305 kg) and used in a 2 x 2 factorial arrangement with main effects of bST (0 or 33 microg x kg BW(-1) x d(-1)) and level of feed intake (ad libitum [AL] or 0.75 AL). Relative to uninjected cattle, treatment with bST increased ADG 9.6% (1.14 vs 1.25 kg/d; P < 0.05) and increased G:F 8.1% (12.3 vs 13.3 gain [g]:feed [kg]; P < 0.05), whereas ADG in AL animals was 39% greater than that in 0.75 AL animals (1.39 vs 1.00 kg/d; P < 0.05). There was a tendency (P = 0.10) for a bST x level of feeding interaction, such that the increase in ADG with bST was greater in AL cattle than in 0.75 AL cattle (10.6 vs 7.8%; P = 0.10). Serum concentrations of ST were greater in 0.75 AL cattle than in AL cattle (13.0 vs 8.6 ng/mL; P < 0.05) and in bST-treated cattle than in uninjected cattle (16.3 vs 5.2 ng/mL; P < 0.05). Due to a bST x level of feeding interaction (P < 0.01), the magnitude of the increase in serum ST to exogenous bST was greater (P < 0.01) in 0.75 AL cattle than in AL cattle. Relative to uninjected cattle, treatment with bST increased (P < 0.05) serum concentrations of IGF-I and IGFBP-3 and reduced (P < 0.05) concentrations of IGFBP-2. Similarly, AL cattle had greater (P < 0.05) serum concentrations of IGF-I and IGFBP-3 and reduced (P < 0.05) IGFBP-2 compared with 0.75 AL cattle. In summary, treatment with bST increased growth rate and G:F and stimulated serum IGF-I and IGFBP-3 while reducing IGFBP-2. Feeding at 0.75 ad libitum intake reduced the magnitude of response for each of these variables. Thus, limit-feeding may reduce the effect of exogenous bST on growth rate by blunting bST-induced increases in IGF-I and IGFBP-3 and bST-induced decreases in IGFBP-2.

Effect of dietary α-tocopherol supplementation on color and lipid stability in pork.

Myoglobin and lipid oxidation are major causes of quality deterioration in fresh pork. A process to enhance color and lipid stability would prove valuable to the pork industry given the current trend of centralized packaging and distribution to retail markets. Our objective was to determine the effects of dietary α-tocopherol (α-Toc) supplementation on color and lipid stability in ground pork, and loin chops stored in modified atmosphere packaging (MAP). Yorkshire crossbred pigs (n=20) were randomized into two groups and fed diets containing 48 (CON) or 170 mg α-Toc acetate/kg feed (VIT-E) for 6 weeks before slaughter. Plasma α-Toc concentration was measured weekly. Post-slaughter, Boston butt shoulders were ground, formed into patties with or without 1.5% salt, and stored fresh at 4°C for 0, 2, 4, or 6 days, and frozen at -20°C for 45 or 90 days. Pork loin chops were packaged aerobically and stored at 4°C for 0, 2, 4 or 6 days, or in MAP at 4°C for 7, 10 or 13 days prior to Hunter L*,a*,b* and TBARS analyses. α-Toc concentration of longissimus dorsi, psoas major, biceps femoris, semimembranosus and semitendinosus muscles was determined. Plasma α-Toc was greater (P<0.05) in VIT-E animals compared with CON and α-Toc concentrations were greater (P<0.05) in all VIT-E muscles compared with CON. TBARS values of both fresh and salted patties were less in VIT-E than in CON meat following 6 days at 4°C; VIT-E TBARS of salted patties were less (P<0.05) after 45 days at -20°C compared with CON. α-Toc supplementation did not influence (P>0.05) color of aerobically packaged or MAP chops, or of fresh or salted pork patties. α-Toc supplementation reduced TBARS formation in fresh and salted pork but had no significant impact on color.

Poor maternal nutrition during gestation in sheep reduces circulating concentrations of insulin-like growth factor-I and insulin-like growth factor binding protein-3 in offspring.

To determine if poor maternal nutrition alters growth, body composition, circulating growth factors, and expression of genes involved in the development of muscle and adipose of offspring, 24 Dorset and Shropshire ewes were fed either 100% (control fed), 60% (restricted fed), or 126% (over fed) of National Research Council requirements. Diets began at day 116 ± 6 of gestation until parturition. At parturition, 1 lamb from each control fed (CON), restricted fed (RES), and over fed (OVER) ewe was necropsied within 24 h of birth (1 d; n = 3/treatment) or reared on a control diet for 3 mo (CON = 5, RES = 5, and OVER = 3/treatment) and then euthanized. Body weights and blood samples were collected from lambs from 1 d to 3 mo. Organ weights, back fat thickness, loin eye area, and tissue samples (quadriceps, adipose, and liver) were collected at 1 d and 3 mo of age. The RES lambs weighed 16% less than CON (P = 0.01) between 1 d and 3 mo of age. In RES, there was a tendency for reduced heart girth at 1 d and 3 mo (P < 0.07) and back fat was reduced 36% at 3 mo (P = 0.03). Heart weight was 30% greater in OVER at 1 d when compared with RES lambs (P = 0.02). Serum IGF-I and IGFBP-3 were reduced in RES and OVER lambs (P < 0.05). Leptin tended to be greater in OVER lambs compared with CON at 1 d and 3 mo (P ≤ 0.08). Triiodothyronine was reduced in RES at 1 d (P = 0.05) and triglycerides tended to be greater in OVER at 3 mo (P = 0.07). In liver, there was a tendency for increased expression of IGF-I in OVER (P = 0.06) and decreased IGFBP-3 in RES (P = 0.09) compared with CON lambs at 1 d. In adipose tissue, adiponectin expression was decreased in RES (P = 0.05) at 3 mo. At 1 d of age, muscle expression of IGF-I tended to increase in RES (P = 0.06). In conclusion, poor maternal nutrition during gestation reduced growth rate in offspring which may be because of reduced circulating IGF-I and IGFBP-3 and decreased expression of IGFBP-3 in the liver.