Search

Objective With increases in the global population, there is a need to identify strategies that increase beef output while maintaining or improving health benefits of beef products. Studies have demonstrated that there are many benefits to human health in response to reducing the dietary ratio of n-6 to n-3. The aim of this study was to characterize the carcass characteristics and brisket muscle fatty acid profile of young dairy-bred bulls following dietary supplementation with n-6 or n-3 PUFA. Materials and Methods Holstein-Friesian (n = 43) and Jersey (n = 7) bulls with a mean ± SEM age and BW of 420.1 ± 5.86 d and 382.0 ± 8.94 kg, respectively, were offered a cereal-based concentrate diet on an ad libitum basis, fortified with 1 of 3 lipid supplements: control (CTL; no supplementary lipid), n-6 PUFA safflower oil (SO), or n-3 PUFA enriched fish oil (FO). Bulls were individually offered their respective diet for 12 wk before slaughter. Carcass weight, conformation, and fat score were recorded at slaughter for all animals. Brisket muscle was collected from 26 randomly selected bulls, and lipid profile was analyzed using gas chromatography. Results and Discussion Total n-3 PUFA concentration was greater for FO than for either SO or CTL diets (P Implications and Applications Beef producers can feed diets enriched with n-6 and n-3 PUFA, which would result in beef having potential health benefits and greater branding potential.

The relationship between rumen microbiota and host feed efficiency phenotype, for genetically divergent beef cattle breeds is unclear. This is further exacerbated when different growth stages, chemically diverse diets and production systems are considered. Residual feed intake (RFI), a measure of feed efficiency, was calculated for individually fed Charolais (CH) and Holstein–Friesian (HF) steers during each of four 70-day (excluding adaptation) successive dietary phases: namely, high-concentrate, grass silage, fresh zero-grazed grass and high-concentrate again. Rumen fluid from the ten highest- (HRFI) and ten lowest-ranking (LRFI) animals for RFI, within breed, during each dietary phase was collected using a trans-oesophageal sampler and subjected to 16S rRNA amplicon sequencing and metabolic profiling. The datasets were analysed to identify microbial and rumen fermentation markers associated with RFI status. Age, dietary phase and breed were included in the statistical model. Within breed, for each dietary phase, mid-test metabolic weight and average daily gain did not differ (P > 0.05) between HRFI and LRFI steers; however, for the initial high-concentrate, grass silage, fresh grass herbage and final high-concentrate dietary phases, HRFI HF steers consumed 19, 23, 18 and 27% more (P Mogibacteriaceae and the genus p-75-a5 of Erysipelotrichaceae and were negatively associated (q

Younger bulls typically produce lower volumes of semen per ejaculate with a lower sperm concentration than older more mature, bulls and often fail to meet semen demand using standard collection frequency schedules. The objective of this study was to assess the effect of ejaculate collection frequency on semen output, sperm quality and field fertility in young bulls under commercial conditions. Holstein Friesian bulls aged 366 ± 8 days (mean ± SEM) were assigned one of two ejaculate collection frequencies: (i) HF (n = 14 bulls), where ejaculates were collected twice a day, five days in each two-week period or (ii) LF (n = 12 bulls), where ejaculates were collected once a day, two days per week. The trial period continued until each bull reached both 20 ejaculates and 1000 marketable frozen semen straws. Subjective motility was assessed on all ejaculates pre-freeze and post-thaw (at 0 and 2 h). A subset of ejaculates were assessed post-thaw by computer-assisted sperm analysis for motility, kinematics and morphological defects and by flow cytometry for viability, membrane fluidity, acrosome integrity, reactive oxygen species and DNA fragmentation. A total of 13,846 inseminations (9,541 for HF and 4,305 for LF) were carried out on dairy cows and heifers. HF reached the 1000 straw threshold 41 days earlier than LF (P 0.01) with the same number of ejaculates. Ejaculate volume and sperm concentration were not affected by treatment but the first ejaculate of the day (HF only) had a greater volume (P 0.001) and sperm concentration (P 0.05) than the second ejaculate. HF had higher pre-freeze total (P 0.01) and gross (P 0.05) motility than LF. HF had higher post-thaw (2 h) total and gross motility than LF (P 0.05). Ejaculate rejection rates did not differ between treatments. There was no effect of treatment, week or ejaculate number of the day (HF only) on post-thaw motility and kinematic parameters or sperm viability, membrane fluidity, acrosome integrity and DNA fragmentation. However, HF had lower superoxide production than LF (P 0.05). Pregnancy per artificial insemination was 64.5 ± 1.0% and 59.9 ± 1.1% for the HF and LF bulls, respectively (mean ± SEM; P = 0.05). In conclusion, collecting ejaculates more frequently from young bulls significantly reduced the number of days required to obtain 1000 straws, increased semen quality in terms of lower superoxide production and increased field fertility.

The objective of this study was to examine the effect of nutrition during the first 12 wk of life on aspects of the physiological and transcriptional regulation of testicular and overall sexual development in the bull calf. Holstein Friesian bull calves with a mean (SD) age and bodyweight of 17.5 (2.85) d and 48.8 (5.30) kg, respectively, were assigned to either a high (HI; n = 15) or moderate (MOD; n = 15) plane of nutrition and were individually fed milk replacer and concentrate to achieve overall target growth rates of at least 1.0 and 0.5 kg/d, respectively. Throughout the trial, animal growth performance, feed intake, and systemic concentrations of metabolites, metabolic hormones, and reproductive hormones were assessed. Additionally, pulsatility of reproductive hormones (luteinizing hormone, follicle-stimulating hormone, and testosterone) was recorded at 15-min intervals during a 10-h period at 10 wk of age. At 87 ± 2.14 d of age, all calves were euthanized, testes were weighed, and testicular tissue was harvested. Differential expression of messenger ribonucleic acid (mRNA) candidate genes involved in testicular development was examined using quantitative polymerase chain reaction assays. All data were analyzed using the MIXED procedure in Statistical Analysis Software using terms for treatment as well as time for repeated measures. Blood metabolites and metabolic hormones generally reflected the improved metabolic status of the calves on the HI plane of nutrition though the concentrations of reproductive hormones were not affected by diet. Calves on the HI diet had greater mean (SED) slaughter weight (112.4 vs. 87.70 [2.98] kg; P < 0.0001) and testicular tissue weight (29.2 vs. 20.1 [2.21] g; P = 0.0003) than those on the MOD diet. Relative mRNA abundance data indicated advanced testicular development through upregulation of genes involved in cellular metabolism (SIRT1; P = 0.0282), cholesterol biosynthesis (EBP; P = 0.007), testicular function (INSL3; P = 0.0077), and Sertoli cell development (CLDN11; P = 0.0054) in HI compared with MOD calves. In conclusion, results demonstrate that offering dairy-bred male calves a high plane of nutrition during the first 3 mo of life not only improves growth performance and metabolic status but also advances testicular development consistent with more precocious sexual maturation.

Increasing the content of polyunsaturated fat in the human diet is a priority for reducing cardiovascular disease and cancer risks. Beef has the potential to contribute to the polyunsaturated fat content in the human diet; however, ruminants cannot synthesise many long-chain fatty acids de novo; they require dietary supplementation. The objectives of the current study were to evaluate (i) the effect of a partially rumen protected n-3 long-chain polyunsaturated fatty acid (LC-PUFA) dietary supplement on the fatty acid composition of muscle (Longissimus dorsi), adipose and liver tissues of beef heifers and (ii) the usefulness of blood plasma as a predictor of tissue concentrations of specific fatty acids. Charolais crossbred heifers were assigned to one of two isolipid dietary treatments (n = 10), namely palmitic acid (control) or an n-3 LC-PUFA supplement for a 91-day period. Blood plasma and adipose tissue samples were taken to determine the temporal effect of these diets on fatty acid composition (days 0, 10, 35 and 91), while liver and muscle samples were taken following slaughter. Dietary lipid source did not influence animal growth rate or body condition score. At day 91, the percentage differences between control and n-3 LC-PUFA heifers in concentrations of eicosapentaenoic acid were +61, +176 and +133 % in liver, muscle and adipose, respectively. For docosahexaenoic acid, at the same time point, the percentage differences were +57, +73 and +138 % for liver, muscle and adipose, respectively. Medium-to-strong positive correlation coefficients were evident for liver and plasma fatty acids, in particular, there were positive relationships with concentrations of total saturated fatty acid (SFA), total n-6 PUFA and total n-3 PUFA. This trend also extended to both the ratio of PUFA to SFA (slope (β1) = 0.56 ± 0.167, intercept (β0) = 0.56, R2 = 0.61, P < 0.05) and the ratio of n-6 to n-3 PUFA (β1 = 0.15 ± 0.054, β0 = 0.24, R2 = 0.52, P < 0.05). A strong correlation was also detected in the ratio of n-6 to n-3 in plasma and muscle tissue of heifers fed the n-3 LC-PUFA diet (β1 = 0.53 ± 0.089, β0 = -0.31, R2 = 0.83, P < 0.001). The results of this study show that the n-3 LC-PUFA can be readily increased through targeted supplementation and that plasma concentrations of n-3 LC-PUFA are useful predictors of their concentrations in a number of economically important tissues.

The aim was to examine the effect of rapid body weight gain during early calfhood consistent with earlier sexual development on the transcriptional profile of the hypothalamus. Angus X Holstein-Friesian heifer calves (19 ± 5 days of age) were offered a high (HI, n = 14) or moderate (MOD, n = 15) plane of nutrition from 3 to 21 weeks of age to achieve a growth rate of 1.2 kg/d and 0.5 kg/d, respectively. Following euthanasia at 21 weeks, the arcuate nucleus (ARC) region was separated from the remainder of the hypothalamus and both were subjected to RNA-Seq. HI calves exhibited altered expression of 80 and 39 transcripts in the ARC and the remaining hypothalamus, respectively (P, The authors would like to acknowledge all skilled technical assistance and support from the Farm Manager and Farm Staf from Teagasc, Grange Beef Research Centre (Dunsany, Co. Meath, Ireland) during the course it this study. Tis research was funded by the Department of Agriculture, Food & Marine, Ireland through the Research Stimulus Fund (13/S/515)

This study examined the effect of plane of nutrition on the endocrinological regulation of the hypothalamic–pituitary–ovarian (HPO) axis in beef heifer calves during a critical sexual developmental window early in calf hood. Forty Holstein-Friesian × Angus heifers (mean age 19 d, SEM = 0.63) were assigned to a high (HI; ADG 1.2 kg) or moderate (MOD; ADG 0.50 kg) nutritional level from 3 to 21 wk of life. Intake was recorded using an electronic calf feeding system, BW was recorded weekly, and blood samples were collected on the week of age 5, 10, 15, and 20 for metabolite, reproductive, and metabolic hormone determination. At 19 wk of age, on sequential days, an 8-h window bleed was carried out for luteinizing hormone (LH), follicle-stimulating hormone (FSH), and estradiol analysis. To characterize anterior pituitary gland function, an intravenous GnRH challenge was conducted (19 wk of age). Blood was collected via a jugular catheter every 15 min for 135 min for the analysis of LH, FSH, and estradiol. Calves were subsequently euthanized at 21 wk of age; the anterior pituitary, metabolic organs, and reproductive tract were weighed, and ovarian surface follicular numbers and oocytes recovered were recorded. Mean ADG was 1.18 and 0.50 kg for HI and MOD, respectively, resulting in a 76.6-kg difference in BW (P < 0.001). Blood insulin, glucose, and IGF-1 concentrations were greater (P < 0.001) for HI compared with MOD. There was a diet × time interaction for leptin (P < 0.01); concentrations were greater in HI compared with MOD at 20 wk of age with no difference between treatments before this. Dietary treatment did not alter the concentrations of adiponectin or anti-mullerian hormone. There was a diet × time interaction for FSH, whereby MOD had greater concentrations than HI at 10, 15, and 20, but not at 5 wk of age. Over the duration of an 8-h window bleed (19 wk of age), serum concentrations of LH, LH pulse frequency, and LH pulse amplitude were unaffected by treatment, whereas FSH (0.23 vs. 0.43 ng/mL) and estradiol (0.53 vs. 0.38 ng/mL) concentrations were less than and greater, respectively, for HI than MOD (P < 0.05). Likewise, following a GnRH challenge, the area under the curve analysis revealed greater (P < 0.01) estradiol and lesser (P < 0.01) FSH concentrations in calves on the HI relative to MOD diet, whereas concentrations of LH were unaffected (P = 0.26) between treatments. Ovarian surface follicle numbers were greater (P < 0.05) in HI compared with MOD. Total reproductive tract, uterus, and ovarian tissue expressed relative to BW were greater (P < 0.05) for HI compared with MOD. In conclusion, enhanced nutrition in early calfhood advances the ontogeny development of the HPO axis.

The advent of genomic selection has accentuated interest in procuring saleable semen from young genetically elite bulls as early in life as possible. However, the timing of availability of semen for commercial use will be determined by the age at which these young animals reach puberty and subsequent sexual maturity. Enhancing early-life nutrition stimulates the functionality of the hypothalamic–pituitary–testicular axis, mediated through complex biochemical interplay between metabolic and neuroendocrine signals and culminating in enhanced testicular growth, steroidogenesis, spermatogenesis, and ultimately, earlier onset of sexual maturation. Indeed, recent evidence indicates that the timing of sexual precocity, which is dictated by an early gonadotropin rise (8–20 wk of age) in the bull, is determined by prevailing metabolic status during calfhood and is not compensable even where prior undernutrition is followed by dietary augmentation. However, despite this, the precise neuronal mechanisms regulating these developmental processes remain to be elucidated for the bull. Although early-life nutrition clearly affects the sexual maturation process, there is little evidence for latent effects on postpubertal semen characteristics. Equally, postpubertal fertility, measured as in vitro fertilization and early embryogenesis, is not influenced by nutritional status during early life. Current efforts employing high throughput nucleic acid and proteomic sequencing and targeted immunofluorescence coupled with systems biology–based gene network analyses can provide further insight on how nutrition may mediate the biochemical interaction between neuroendocrine and testicular cellular processes. Such information can be harnessed to identify potential genomic targets as part of genomically assisted breeding programs as well as facilitate development of strategic nutritional regimens to optimize sexual maturation and subsequent semen availability from genetically elite young bulls.

The aim of this study was to investigate the effect of early calf-hood nutrition on the transcriptomic profile of the arcuate nucleus of the hypothalamus, anterior pituitary and testes in Holstein-Friesian bulls. Holstein-Friesian bull calves with a mean (±S.D.) age and bodyweight of 19 (±8.2) days and 47.5 (±5.3) kg, respectively, were offered a high (n = 10) or low (n = 10) plane of nutrition in order to achieve an overall growth rate of 1.2 and 0.5 kg/day. At 126 (±3) days of age, calves were euthanized, hypothalamus (arcuate region), anterior pituitary and testicular parenchyma samples were harvested and RNAseq analysis was performed. There were 0, 49 and 1,346 genes differentially expressed in the arcuate nucleus, anterior pituitary and testicular tissue of bull calves on the low relative to the high plane of nutrition, respectively (P

peer-reviewed In sheep, cervical artificial insemination (AI) involves depositing semen at the cervical opening, as it is not possible to traverse the cervix due to its complex anatomy. However, internationally this method yields low pregnancy rates when frozen-thawed semen is used. An exception to this is in Norway, in which vaginal deposition of frozen-thawed semen to a natural estrus yields pregnancy rates around 70%. As the cervix and its secretions are the principal factors influencing sperm transport to the site of fertilization the aim of this study was to characterise the differences in the cervical anatomy as well as the cervicovaginal mucus properties of six European ewe breeds across three countries known to have differences in pregnancy rates following cervical AI with frozen-thawed semen. These were Suffolk and Belclare in Ireland, Fur and Norwegian White Sheep (NWS) in Norway and Ile de France and Romanov in France (n ¼ 28e30 ewes/breed). Cervicovaginal mucus was collected at the follicular and luteal phases of both a synchronized and natural cycle and assessed for mucus weight, viscosity and colour. The anatomical characteristics of the cervix (length of the cervix, number of cervical rings and the appearance of the external os) were assessed post-mortem. There was a type of the cycle by ewe breed interaction represented by no differences in mucus production between ewe breeds at the natural cycle for both the follicular and luteal phases of the cycle. However, there were differences between ewe breeds at the synchronized cycle (P < 0.05). Belclare had the lowest mucus production at the follicular phase while NWS had the lowest amount of mucus at the luteal phase of the synchronized cycle. Overall, across all ewe breeds, mucus production was higher at the follicular than at the luteal phase (P < 0.05). Despite reports of Suffolk and NWS having the most divergent pregnancy rates following cervical AI with frozen-thawed semen, both breeds had the lowest overall mucus viscosity at the follicular phase of both types of cycle with no differences between both ewe breeds (P > 0.05). The length of the cervix, number of cervical rings and the external os type were affected by ewe breed (P < 0.05). Suffolk ewes had longer cervices but lower number of cervical rings than NWS and Fur ewes (both with higher pregnancy rates). In conclusion, while mucus production and mucus viscosity was affected by breed, these changes are not consistent with the known differences between ewe breeds in their pregnancy rates following cervical AI with frozen-thawed semen. PUBLISHED peer-reviewed

Manipulation of the rumen microorganisms during early life has emerged as a promising strategy for persistent improvement of nutrient utilisation and lowering of enteric methanogenesis. However, limited understanding of the dynamics of rumen microbial colonisation has prevented the identification of the optimum timeframe for such interventions. The present study used DNA amplicon sequencing of the 16S rRNA gene to assess bacterial and archaeal dynamics in the rumen digesta of beef calves raised on two farms from birth through to post-weaning. The colonisation patterns of both communities were influenced by age (P < 0.05) and farm of origin (P < 0.05). The bacterial community exhibited an age-wise progression during the first month of life which appeared to be partly related to diet, and settled by day 21, indicating that this may mark the boundary of a timeframe for intervention. The archaeal community appeared less sensitive to age/diet than bacteria in the first month of life but was more sensitive to farm environment. These data show that ruminal microbial composition during early life is driven by calf age, diet and local environment, and provide important fundamental information concerning the ontogeny of the rumen microbiota from birth.

Early-life nutrition affects calf development and thus subsequent performance. The aim of this study was to examine the effect plane of nutrition on growth, feeding behaviour and systemic metabolite concentrations of artificially reared dairy bull calves. Holstein-Friesian (F; n=42) and Jersey (J; n=25) bull calves with a mean±SD age (14±4.7 v. 27±7.2 days) and BW (47±5.5 v. 33±4.7 kg) were offered a high, medium or low plane of nutrition for 8 weeks using an electronic feeding system which recorded a range of feed-related events. Calves were weighed weekly and plasma samples were collected via jugular venipuncture on weeks 1, 4 and 7 relative to the start of the trial period. The calves offered a high plane of nutrition had the greatest growth rate. However, the increased consumption of milk replacer led to a reduction in feed efficiency. Holstein-Friesian calves offered a low plane of nutrition had the greatest number of daily unrewarded visits to the feeder (P

Early onset of puberty is a key trait underpinning economically efficient cattle production systems. Although prepubertal plane of nutrition has been positively associated with earlier onset of puberty, the underlying biochemical mechanisms remain unknown. The hypothalamus, and particularly the arcuate nucleus (ARC), has a pivotal role during this developmental stage because it is a central regulator of homeostasis, mediating various neuroendocrine and physiological functions, including feed intake and reproduction. Thus, we hypothesised that a high plane of nutrition during early calfhood would affect the transcriptional profile of hypothalamic tissue, and in particular, elicit differential expression of biochemical pathways consistent with earlier sexual development in beef heifers. Angus×Holstein-Friesian heifer calves (19±5 days of age, 51.2±7.8kg, mean±s.d.) were offered a high (HP, n=14) or moderate plane of nutrition (MP, n=15) from 3 to 21 weeks of age to achieve target growth rates of 1.2 and 0.5 kg/d, respectively. At 21 weeks of age, calves were killed and the brain recovered. The ARC region was separated from the remainder of the hypothalamus, with both tissues flash frozen and subsequently used for RNA sequencing. Differential expression of genes (DEGs) was determining using R Bioconductor package EdgeR (version 3.20.9). The resultant list of DEGs was then submitted to Ingenuity Pathway Analysis and Weighted Gene Co-expression Network Analysis (WGCNA). At slaughter, bodyweight was higher in HP calves (189.6 vs. 113.0 kg; P

peer-reviewed Leydig cell functional capacity reflects the numbers and differentiation status of the steroidogenic Leydig cells in the testes and becomes more or less fixed in early adulthood with the final establishment of the hypothalamo-pituitary-gonadal (HPG) axis after puberty. Factors influencing Leydig cell functional capacity and its role in puberty are poorly understood. Using a bovine model of dairy bulls fed four different nutritional regimes from 1 month to 12 months, and applying circulating Insulin-like peptide 3 (INSL3) as an accurate biomarker of Leydig cell functional capacity, showed that a high plane of nutrition in the first 6 months of life, but not later, significantly increased INSL3 in young adulthood. Moreover, INSL3 concentration at 4 months indicated a marked differential in early feeding regime and correlated well (negatively) with the timing of puberty, as reflected by the age in days for the first production of an ejaculate with >50 million sperm and >10% forward motility, as well as with testis size at 18 months. Reversing the diet at 6 months was unable to rectify the trend in either parameter, unlike for other parameters such as testosterone, body weight, and scrotal circumference. This study has shown that early prepubertal nutrition is a key factor in the development of Leydig cell functional capacity in early adulthood and appears to be a key driver in the dynamic progression of puberty.

The objective of this study was to examine the effect of nutrition during the first 18 weeks of life on the physiological and transcriptional functionality of the hypothalamic (arcuate nucleus region), anterior pituitary and testes in Holstein–Friesian bull calves. Holstein–Friesian bull calves with a mean (±S.D.) age and bodyweight of 19 (±8.2) days and 47.5 (±5.3) kg, respectively, were assigned to either a HIGH (n = 10) or LOW (n = 10) plane of nutrition, to achieve an overall target growth rate of 1.2 or 0.5 kg/day, respectively. At 126 ± 1.1 days of age, all calves were euthanised. Animal performance (weekly) and systemic concentrations of metabolic (monthly) and reproductive hormones (fortnightly) were assessed. Testicular histology, targeted gene and protein expression of the arcuate nucleus region, anterior pituitary and testes were also assessed using qPCR and immunohistochemistry, respectively. The expression of candidate genes in testicular tissue from post pubertal 19-month-old Holstein–Friesian bulls (n = 10) was compared to that of the 18-week-old calves. Metabolite and metabolic hormone profiles generally reflected the improved metabolic status of the calves on the HIGH (P< 0.001). Calves offered a HIGH plane of nutrition were heavier at slaughter (P < 0.001), had larger testes (P < 0.001), larger seminiferous tubule diameter (P < 0.001), more mature spermatogenic cells (P < 0.001) and more Sertoli cells (P < 0.05) in accordance with both morphological and transcriptional data. Overall, testicular gene expression profiles suggested a more mature stage of development in HIGH compared with LOW and were more closely aligned to that of mature bulls. Ghrelin receptor was the only differentially expressed gene between LOW and HIGH calves in either the anterior pituitary (P < 0.05) or arcuate nucleus region of the hypothalamus (P < 0.10) and was upregulated in LOW for both tissues. This study indicates that an enhanced plane of nutrition during early calfhood favourably alters the biochemical regulation of the hypothalamus–anterior pituitary–testicular axis, advancing testicular development and hastening spermatogenesis.

peer-reviewed The dataset generated during and analysed in the current study are available in the NCBI’s Gene Expression Omnibus repository, and are accessible through GEO Series accession number GSE97674 [https://www.ncbi.nlm.nih.gov/gds/?term=GSE97674]. Background Adipose tissue is a major endocrine organ and is thought to play a central role in the metabolic control of reproductive function in cattle. Plane of nutrition during early life has been shown to influence the timing of puberty in both male and female cattle, though the exact biological mechanisms involved are currently unknown. The aim of this study was to investigate the effect of early calf-hood nutrition on the transcriptomic profile of subcutaneous adipose tissue in Holstein-Friesian bulls to identify possible downstream effects on reproductive physiology. Results Holstein-Friesian bull calves with a mean (±S.D.) age and bodyweight of 19 (±8.2) days and 47.5 (±5.3) kg, respectively, were assigned to either a high (n = 10) or low (n = 10) plane of nutrition. Calves were fed in order to achieve an overall growth rate of 1.08 and 0.57 kg/day for the high and low plane of nutrition treatments, respectively. At 126 days of age, the bulls were euthanized, subcutaneous adipose tissue samples were harvested and RNAseq analysis was performed. There were 674 genes differentially expressed in adipose tissue of calves on the low compared with the high plane of nutrition (P 2.0). High plane of nutrition positively altered the expression of genes across an array of putative biological processes but the most dominant cellular processes affected were cellular energy production and branched chain amino acid degradation. A high plane of nutrition caused upregulation of genes such as leptin (LEP) and adiponectin (ADIPOQ), which are known to directly affect reproductive function. Conclusions These results provide an insight into the effect of augmenting the plane of nutrition of Holstein-Friesian bull calves in the prepubertal period on the transcriptome of adipose tissue.

peer-reviewed The aim of this study was (1) to examine the effect of plane of nutrition during the first and second 6 mo of life on systemic concentrations of reproductive hormones and metabolites in Holstein-Friesian dairy bulls, and (2) to establish relationships with age at puberty and postpubertal semen production potential. Holstein-Friesian bull calves (n = 83) with a mean (standard deviation) age and body weight of 17 (4.4) d and 52 (6.2) kg, respectively, were assigned to a high or low plane of nutrition for the first 6 mo of life. At 24 wk of age, bulls were reassigned, within treatment, either to remain on the same diet or to switch to the opposite diet until puberty, resulting in 4 treatment groups: high-high, high-low, low-low, and low-high. Monthly blood samples were analyzed for metabolites (albumin, urea, total protein, β-hydroxybutyrate, glucose, nonesterified fatty acid, triglycerides and creatinine), insulin, insulin-like growth factor-1, leptin, adiponectin, FSH, and testosterone. A GnRH challenge was carried out at 16 and 32 wk of age (n = 9 bulls per treatment). Blood was collected at 15-min intervals for 165 min, with GnRH administered (0.05 mg/kg of body weight, i.v.) immediately after the third blood sample. Blood samples were subsequently analyzed for LH, FSH, and testosterone. Stepwise regression was used to detect growth and blood measurements to identify putative predictors of age at puberty and subsequent semen quality traits. Metabolic hormones and metabolites, in general, reflected metabolic status of bulls. Although FSH was unaffected by diet, it decreased with age both in monthly samples and following GnRH administration. Testosterone was greater in bulls on the high diet before and after 6 mo of age. Testosterone concentrations increased dramatically after 6 mo of age. Luteinizing hormone was unaffected by diet following GnRH administration but basal serum LH was greater in bulls on a high diet before 6 mo of age. In conclusion, the plane of nutrition offered before 6 mo of age influenced metabolic profiles, which are important for promoting GnRH pulsatility, in young bulls

The advent of genomic selection has led to increased interest within the cattle breeding industry to market semen from young bulls as early as possible. However, both the quantity and quality of such semen is dictated by the age at which these animals reach puberty. Enhancing early life plane of nutrition of the bull stimulates a complex biochemical interplay involving metabolic and neuroendocrine signalling and culminating in enhanced testicular growth and development and earlier onset of sexual maturation. Recent evidence suggests that an enhanced plane of nutrition leads to an advancement of testicular development in bulls at 18 weeks of age. However, as of yet, much of the neuronal mechanisms regulating these developmental processes remain to be elucidated in the bull. While early life nutrition clearly affects the sexual maturation process in bulls, there is little evidence for latent effects on semen traits post-puberty. Equally the influence of prevailing nutritional status on the fertility of mature bulls is unclear though management practices that result in clinical or even subclinical metabolic disease can undoubtedly impact upon normal sexual function. Dietary supplements enriched with various polyunsaturated fatty acids or fortified with trace elements do not consistently affect reproductive function in the bull, certainly where animals are already adequately nourished. Further insight on how nutrition mediates the biochemical interaction between neuroendocrine and testicular processes will facilitate optimisation of nutritional regimens to optimise sexual maturation and subsequent semen production in bulls.

The aim of this study was to examine the effects of dietary supplementation with rumen protected n-6 or n-3 polyunsaturated fatty acids (PUFA) on the quantity and quality of semen from young post-pubertal dairy bulls. Pubertal Holstein-Friesian (n=43) and Jersey (n=7) bulls with a mean±s.e.m. age and bodyweight of 420.1±5.86 days and 382±8.94kg, respectively, were blocked on breed, weight, age and semen quality (based on the outcomes of two pre-trial ejaculates) and randomly assigned to one of three treatments: (i) a non-supplemented control (CTL, n=15), (ii) rumen-protected safflower (SO, n=15), (iii) rumen-protected n-3 PUFA-enriched fish oil (FO, n=20). Bulls were fed their respective diets,ad libitumfor 12 weeks; individual intakes were recorded using an electronic feeding system for the initial 6 weeks of the feeding period. Semen was collected via electro-ejaculation at weeks−2,−1, 0, 7, 10, 11 and 12 relative to the beginning of the trial period (week 0). On collection, semen volume, sperm concentration and progressive linear motility (PLM) were assessed. On weeks−2,−1, 0, 10, 11, 12, semen was packaged into 0.25mL straws and frozen using a programmable freezer. On weeks−1, 7 and 11; a sub-sample of semen was separated into sperm and seminal plasma, by centrifugation and stored at – 20°C until analysis of lipid composition. Semen from 10 bulls per treatment were used for post-thaw analysis at weeks 10, 11 and 12 (3 straws per ejaculate). Sperm motility was analysed by computer assisted semen analysis (CASA). In addition, membrane fluidity, acrosome reaction and oxidative stress were assessed using flow cytometry. Sperm from bulls fed SO had a 1.2 fold higher total n-6 PUFA content at week 11 compared to week−1 (P

The onset of puberty in the bull is regulated by the timing of early GnRH pulsatility release from the hypothalamus, which has been demonstrated to be affected by plane of nutrition during calf-hood. The aim of this study was to determine the effect of plane of nutrition on growth rate, scrotal development, metabolite concentrations and exogenous gonadotrophin (GnRH) induced release of luteinizing hormone (LH), follicle stimulating hormone (FSH) and testosterone (TT) in pre-pubertal bulls of two contrasting dairy breeds. Holstein-Friesian and Jersey bull calves were assigned to either a high or low plane of nutrition from 3 to 49 weeks of age. Intensive blood sampling was conducted at 16, 24 and 32 weeks of age, every 15 min from 30 min prior to intravenous administration of exogenous GnRH to 135 min after. Monthly blood samples were also collected and analyzed for insulin like growth factor 1 (IGF-1), insulin, leptin, adiponectin and metabolite concentration. Insulin and IGF-1 were higher in bulls on a high plane of nutrition (P 0.001) but were not affected by breed (P 0.05). Leptin was not affected by plane of nutrition or breed (P 0.05). Adiponectin tended to be higher in bulls on a high plane of nutrition (P = 0.05), but was not affected by breed (P 0.05). Bulls on a high plane of nutrition had a greater concentration of LH in response to GnRH (P 0.05) but there was no effect of breed (P 0.05). FSH concentration was not influenced by breed or plane of nutrition but FSH concentrations did decrease with age (P 0.01), while, LH was not affected by age (P 0.05). Jersey bulls, particularly those on a high plane of nutrition, had higher TT production in the pre-pubertal period (P 0.001). Using 28 cm as a proxy for age at puberty, bulls on a high plane of nutrition were predicted to reach puberty earlier than bulls on a low plane. In conclusion, the data clearly demonstrate that a high plane of nutrition positively affects several key nutritional and reproductive hormones which are critical to the endocrinological functionality of the hypothalamic-pituitary-testicular axis in dairy-bred bull calves.

Oral delivery of gene therapeutics would facilitate treatment of local intestinal disease, including colon cancer and inflammatory bowel disease, thus avoiding invasive surgery. The aims of this study were to investigate; if the orientation of the lipid tail on the cyclodextrin (CD) influenced the efficacy of a novel poly-6-cationic amphiphilic CD to transfect intestinal enterocytes; the endocytotic uptake pathway(s), and, the intracellular trafficking of the CD.DNA complexes. Inhibitors of clathrin- and caveolae- mediated endocytosis and macropinocytosis were used to determine the mechanism(s) of CD.DNA uptake by both undifferentiated and differentiated Caco-2 cells. Cell surface heparan sulphate proteoglycans were involved in the association of CD.DNA complexes with undifferentiated Caco-2 cells. Complexation of pDNA with CD facilitated significant levels of pDNA uptake and gene expression (comparable to PEI) in both undifferentiated and differentiated Caco-2 cells. Disruption of intracellular vesicular trafficking reduced transfection activity. CD was also capable of transfecting the more physiologically relevant differentiated Caco-2 model. Macropinocytosis was responsible for the uptake of CD.DNA transfection complexes by both undifferentiated and differentiated Caco-2 cells. The ability of this novel CD to transfect differentiated intestinal cells indicates the potential of this vector for oral gene delivery.

Early-life nutrition is reported to increase reproductive performance in cattle. The aim of this study was to examine the effect of enhanced nutrition during the first 21 weeks of life on reproductive and metabolic organ growth in heifers. Holstein-FriesianXAngus heifer calves with a mean (±S.D.) age and bodyweight of 19(±5) days and 51.2(±7.8) kg, respectively, were assigned to either a high-energy diet (HE;n = 16; targeted ADG 1.2kg/d) or a control diet (CONT;n = 16; targeted ADG 0.50kg/d). At 145 ± 3 days of age, calves were euthanized and the reproductive tract was excised, dissected (into uterus, cervix and ovary) and weighed together with the anterior-pituitary, liver, heart, lungs, and gastrointestinal tract (GIT). Calves on the HE diet were 189.6kg and CONT diet were 110.2kg, resulting in a 78.4kg weight differential (P < 0.001) at slaughter between the treatment groups. Calves on HE growth was 1.18kg/day and their contemporizes on the CONT achieved 0.502kg/day (P < 0.001). Calves offered the HE diet had greater (P < 0.001) total reproductive tract weight, along with cervix, uterus and ovarian tissue compared CONT calves. Total reproductive tract, cervix, uterus and ovarian tissue when expressed on a body weight basis remained different (P < 0.001) between dietary treatments. Metabolic organs viz heart, lungs, liver, GIT along with the anterior pituitary were greater for calves offered the HE diet (P < 0.001). Early life growth was positively associated (P < 0.001) with reproductive tract, uterus, and ovarian tissue, along with liver and anterior pituitary weight. Indeed, size of anterior pituitary, the liver and the ovary were all positively related to one another (P < 0.05) on an absolute or bodyweight basis.. In conclusion, enhanced early-life nutrition can increase the size of the reproductive tract and ovarian tissue in young female calves. The significance of these morphology differences to advancing sexual maturation in heifers needs to be elucidate.

Advancing the age at which puberty and subsequent sexual maturation is attained in cattle is central to the reproductive and economic efficiency of both beef and dairy production systems worldwide. Onset of puberty in both male and female cattle is regulated by a complex network of biochemical processes and involves interaction among many key metabolic, neuroendocrine and reproductive tissues. Although our understanding of the biochemical interplay that conditions and eventually triggers the pubertal process has improved in recent years, much of the intricate mechanistic detail still eludes us. Environmental factors, such as nutritional management, as well as the genetic makeup of the animal undoubtedly affect the timing of puberty in cattle. In particular, there is now overwhelming evidence to support the importance of early life nutrition in regulating the timing of puberty in both bulls and heifers. For both genders, there is significant evidence that an improved metabolic status, early in calfhood, advances maturation of the hypothalamic–pituitary–gonadal axis, therefore facilitating earlier sexual development. Although advancing sexual maturation is a desirable goal, it is important that any strategy used does not impinge upon normal gametogenesis or postpubertal fertility potential. To this end, the aim of this review is to discuss the underlying biology of puberty in cattle with particular emphasis on the role of nutritional management during early calfhood in: (1) advancing the maturity of the hypothalamic–pituitary–gonadal axis; and (2) implications for the quality of gametes and subsequent fertility.

Local gene delivery represents a promising therapeutic approach for diseases of the intestine. However, the gastrointestinal tract poses significant challenges to successful gene delivery. Cyclodextrins (CDs) have been extensively investigated as non-viral vectors. Here, we assessed the suitability of an amphiphilic cationic CD for intestinal gene transfer, with particular focus on extracellular barriers. Stability and transfection efficiency of CD·DNA complexes were assessed post incubation in simulated gastric and intestinal fluids, bile salts and mucin, or with intestinal enzymes to represent extracellular barriers to intestinal gene delivery. Stability was determined by gel electrophoresis and transfection was measured by luciferase expression in intestinal epithelial cells (Caco-2). Transfection efficiency of CD·DNA complexes was enhanced after incubation in bile salts but was reduced after incubation in gastric and intestinal fluids and mucin. CD·DNA complexes were stable after incubation with pancreatic enzymes and with a model lower intestinal enzyme. Furthermore, the CD protected pDNA from degradation by DNase. In summary, physiologically relevant in vitro models were established and used to quantify the barriers posed by the intestinal extracellular environment to gene delivery. This systematic assessment identified the advantages and limitations of the CD vector and facilitated the proposal of formulation strategies to overcome these barriers.

The preparation is described of all of the likely dimer and trimer ether-linked components of the anticancer material known as hematoporphyrin derivative. The hydroxyethyl side chain of one porphyrin, related to hematoporphyrin, reacts readily with a bromoethyl system derived from the same, or similar, hematoporphyrin -derived porphyrin to form a diporphyrin ether. The coupling reaction can be controlled to occur in situ, allowing the preparation of a symmetrical ether-linked dimer , or separately to provide an asymmetrically substituted porphyrin dimer . The bromoethyl groups can be formed from either the hydroxyethyl groups or, more slowly, from a vinyl side chain by using hydrogen bromide in dichloromethane. Ether-linked porphyrin trimers can be prepared by reacting a dibromoethyl system with 2 equiv. of a hydroxyethyl -containing porphyrin or by further reaction of an initially formed dimer. Hydroxyethyl side chains in the dimers and trimers are best obtained by reduction of an acetyl group. Some spectral properties of the products are described.

— Spectral properties of several diporphyrin ethers were assessed in different solvents and after accumulation by leukemia L1210 cells in vitro. To facilitate studies in a variety of solvents, both tetramethylesters of the diporphyrin ethers and free acids were employed. For comparison, studies on the corresponding porphyrin monomers were also carried out. The joining of two porphyrins by an ether linkage had several consequences. We observed a blue shift in the Soret band of the ethers, but not of the corresponding simple porphyrins, in protic solvents. This phenomenon is likely related to ether aggregation under conditions which promote H-bonding. The presence of an ether linkage was associated with enhanced fluorescence at 630–640 nm and decreased fluorescence lifetimes and yields, especially in protic solvents. The ether linkage was unaffected by intracellular enzymes, but porphyrin esters were readily hydrolyzed upon accumulation by L1210 cells. The joining of two hematoporphyrin molecules by an ether linkage promoted dye accumulation by L1210 cells. In contrast, accumulation of mesoporphyrin and protoporphyrin was thereby retarded.

The population biology of the fish acanthocephalanAcanthocephalus clavulawas described from 161 wild brown trout,Salmo truttasampled over a two-year period in Clogher Lake in the west of Ireland. Overall prevalence of the parasite was 86% and the mean abundance was 53 worms per fish. Despite the presence of large numbers of worms in the trout very few females (2%) attained full reproductive maturity. This suggests that trout is an accidental host. A sample of yellow eels,Anguilla anguillawas examined at a different time from the same lake. The prevalence ofA. clavulawas 97% and the average abundance was 8 worms per fish. In contrast to the situation in trout, the proportion of female worms attaining full reproductive maturity was 61% fulfilling the expected characteristic of a preferred definitive host. The possible explanations for the very high abundance ofA. clavulain trout are discussed and include the influence of fluctuations in host populations, host diet and the absence of a potential competitor.

The TMN Functional Architecture provides a systematic decomposition of the management functions that telecommunications service suppliers and network operators need to run their enterprise. The TMN Functional Architecture is useful for the following tasks: analysis and description of existing Operations Systems and process flows; specification of new operations systems and process flows; gap analysis of operations plans; planning of migration to new systems and processes; development of new operations systems.

In a multivendor environment, it is necessary to describe both functionality and interfaces of an Operations System (OS) in order to assure interoperability with other systems of a Telecommunications Management Network (TMN). The purpose of this work is to provide a concise way to describe the functionality of OSs, as well as provide guidance to the specification of their interfaces. This paper discusses a methodology by which generic TMN information models may be associated with generic TMN Management Application Functions (MAFs), thereby linking the TMN functional architecture and the TMN information architecture. By "generic information models" we mean object classes that are useful in at least two of the Telecommunications Managed Areas of ITU-T Recommendation M.3100. Once this has been done, the functionality of an OS may be described by listing the generic MAFs it supports, along with the technologies and services it supports. The object classes appropriate to the specific technologies and services supported can be easily found by selecting those that inherit from the generic object classes associated with the supported MAFs. An example applies the method to a set of MAFs concerned with Alarm Surveillance. As a result, the functionality of Alarm Surveillance is precisely defined, in preparation for the design of Operations Systems. The information developed in the figures is captured in a concise table, which identifies the specific generic object classes that are the subject of interactions between specified pairs of MAFs.

This Determination of Importance Evaluation (DIE) applies to the Subsurface Exploratory Studies Facility (ESF), encompassing the Topopah Spring (TS) Loop from Station 0+00 meters (m) at the North Portal to breakthrough at the South Portal (approximately 78+77 m), and ancillary test and operation support areas including the Enhanced Characterization of the Repository Block (ECRB) Cross Drift. This evaluation applies specifically to site characterization testing activities ongoing and planned in the Subsurface ESF. ESF site characterization activities are being performed to obtain the information necessary to determine whether the Yucca Mountain Site is suitable as a geologic repository for spent nuclear fuel and high-level radioactive waste. A more detailed description of these testing activities is provided in Section 6 of this DIE. Generally, the construction and operation of excavations associated with these testing activities are evaluated in the DIE for the Subsurface ESF (CRWMS M&O 1999a) and the DIE for the ESF ECRB Cross Drift (CRWMS M&O 2000a). The scope of this DIE also entails the proposed Unsaturated Zone (UZ) Transport Test at Busted Butte. Although, not a part of the TS Loop or ECRB Cross Drift, the associated testing activities are Subsurface testing activities. Busted Butte is located to the south south-east of the TS Loop and is outside the Conceptual Controlled Area Boundary (CCAB). These activities provide access to the Calico Hills (CH) geologic structure. In the case of Busted Butte, construction and operation of excavations are evaluated herein (since this activity was not previously evaluated in CRWMS M&O 1999a). The objectives of this DIE are to determine whether Subsurface ESF testing, and associated activities, could potentially impact site characterization testing and/or the waste isolation capabilities of the site. Controls needed to limit any potential impacts are identified in Section 13. The validity and veracity of the individual tests, including data collection, are the responsibility of the assigned Principal Investigator(s) (PIS) and are not evaluated in this DIE. This DIE focuses on integrating and compiling the evaluations of previous DIES which were prepared for various ESF subsurface testing activities, including the use of temporary items currently located or being developed for these testing activities, and to provide a bounding evaluation for potential future ESF subsurface testing activities that are sufficiently similar to the generic testing activities addressed herein. Subsurface testing activities items/facilities evaluated herein include: ongoing and planned testing in the TS Loop, alcoves, and niches, planned testing in the ECRB Starter Tunnel, borehole drilling and workover, and tracers, fluids, and materials (TFM) usage. Detailed identification of individual testing items/facilities and generic descriptions for subsurface-testing-related activities are provided in Section 6. The conclusions and requirements of this DIE conservatively bound the conclusions and requirements of previously approved DIES for the ESF subsurface testing activities addressed herein, based on conservative engineering judgment and on concurrence with this DIE (via a formal review process) by the originating and reviewing organizations of the previously approved evaluations. Hence, this DIE supersedes the following DIES listed in Table 1.1.

This DIE applies to the surface facilities component of the Yucca Mountain Site Characterization Project (W) ESF. The ESF complex-including surface and subsurface accommodations--encompasses an area that is approximately six miles wide and nine miles long (approximately 30,000 acres total) (United States Department of Energy [DOE] 1997, p. 9.04). It is located on federally withdrawn lands, near the southwest border of the Nevada Test Site (NTS) in southern Nevada (DOE 1997, p. 9.04). Site characterization activities are conducted within the subsurface ESF to obtain the information necessary to determine whether the Yucca Mountain Site is suitable as a geologic repository for spent nuclear fuel and high-level radioactive waste. Most ESF surface facilities are located within the Conceptual Controlled Area Boundary (CCAB) (DOE 1997, p. 9.04), with the exception of the southeastern most portions of the H-Road and the Water Supply System. Various SBT activities are also conducted throughout the Yucca Mountain region as a part of the overall site-characterization effort. In general, the DIE for SBT Activities (Civilian Radioactive Waste Management System [CRWMS] Management and Operating Contractor [M&O] 1998a) evaluates activities associated with SBT. Potential test-to-test interference and waste isolation impacts associated with SBT activities are also evaluated in CRWMS M&O (1998a).

A new series of amphiphilic cyclodextrins containing cationic groups at the 6-positions and alkyl or biolabile ester groups at the 2-positions has been synthesised. Selective 2-O-allylation followed by photochemical addition of lipophilic thiols made it possible to control lipophilicity in these mesomolecules and allow solubility and self-assembly in water. The cationic groups are cysteamine-derived, while the alkyl and ester groups are C(1)-C(16) and benzyl ester groups. This is a new general synthetic route to a potentially wide range of polycationic cyclodextrins capable of acting as gene delivery vectors by condensing DNA and forming liquid crystalline complexes with oligonucleotides.

The compositions of Photofrin II and haematoporphyrin derivative (HPD) were examined. Dihaematoporphyrin ester was unambiguously synthesized and shown by high performance liquid chromatography (HPLC) not to be a significant component of Photofrin II. Hydrolysis studies showed the presence of an acid-stable oligomer in Photofrin II which comprised about 40% of of an acid-stable oligomer in Photofrin II which comprised about 40% of the total sample. A dimer derivative was isolated from this oligomeric material by fully dehydrating the methyl-esterified acidic hydrolysis product of Photofrin II. The structure of this dimer, in which two porphyrin units are linked by a three-carbon bridge containing a double bond and a methyl group, was determined by nuclear magnetic resonance (NMR) and fast atom bombardment (FAB) mass spectrometry. An analogous trimer was also isolated and identified. Structures for the hydroxyl-containing precursors to the dimer and trimer, and for the acid-stable oligomeric material in Photofrin II, are suggested. The acid-stable oligomeric material is produced in the sodium hydroxide polymerization step of the preparation of Photofrin II.

The synthesis of diporphyrins and analogous compounds related to those present in the oligomeric fraction (Photofrin II) obtained from hematoporphyrin derivative (HPD) is described. The photodynamic activity of these compounds, in vivo, varies from inactive to as active as Photofrin II. Factors that are important in determining this anticancer activity of the synthetic compounds are the presence of hydrophobic side chains, as well as the propionic acid side chains of the hematoporphyrin derived materials, and the nature of the linking group between the porphyrins.

The preparation is described of all of the likely dimer and trimer ether-linked components of the anticancer material known as hematoporphyrin derivative. The hydroxyethyl side chain of one porphyrin, related to hematoporphyrin, reacts readily with a bromoethyl system derived from the same, or similar, hematoporphyrin -derived porphyrin to form a diporphyrin ether. The coupling reaction can be controlled to occur in situ, allowing the preparation of a symmetrical ether-linked dimer , or separately to provide an asymmetrically substituted porphyrin dimer . The bromoethyl groups can be formed from either the hydroxyethyl groups or, more slowly, from a vinyl side chain by using hydrogen bromide in dichloromethane. Ether-linked porphyrin trimers can be prepared by reacting a dibromoethyl system with 2 equiv. of a hydroxyethyl -containing porphyrin or by further reaction of an initially formed dimer. Hydroxyethyl side chains in the dimers and trimers are best obtained by reduction of an acetyl group. Some spectral properties of the products are described.

The presence of carbon linked oligomeric porphyrins in hematoporphyrin derivative and Photofrin II has been established by FAB mass spectrometry and hydrolysis studies. A dimer derived from this material has been isolated and characterized by nmr spectroscopy and mass spectrometry.

In cattle, uterine gland development (adenogenesis) occurs postnatally, beginning immediately after birth, and is normally complete by approximately 2 months of age. Normal development of uterine glands is essential for subsequent fertility. Early life plane of nutrition can impact age at puberty, but the impact of early postnatal nutrition on adenogenesis is unclear. The aim of this study was to examine the effect of enhanced nutrition during the first 21 weeks of life on heifer reproductive organ growth, endometrial gland development, and endometrial gene expression. To this end, Angus × Holstein-Friesian heifer calves with a mean (± s.d.) age and body weight of 19 (± 5) days and 51.2 (± 7.8) kg, respectively, were assigned to either a high (HIGH; n = 16; targeted average daily gain, ADG, 1.2 kg/d) or a moderate plane of nutrition (MOD; n = 16; targeted ADG 0.5 kg/d). Calves were slaughtered at five months of age, and their reproductive tracts were collected and dissected for analysis. To assess endometrial gland number, a cross-section of the midpoint of one horn (n = 8 per group) was fixed in 10% neutral buffered formalin, embedded, and processed for histology. Slides were stained with haematoxylin-eosin. Five pictures were randomly taken on each slide using a stereomicroscope at 40× magnification and the number of glands was counted using ImageJ (https://imagej.nih.gov/ij/) software. In addition, endometrial tissue samples were collected and snap frozen for subsequent quantitative RT-PCR analysis. At the time of slaughter, calves on the HIGH and MOD diets presented a body weight of 189.6 and 110.2 kg, respectively (P