Impacts of chronic and increasing lipopolysaccharide exposure on production and reproductive parameters in lactating Holstein dairy cows.

Lipopolysaccharide (LPS) administration causes immunoactivation, which negatively affects production and fertility, but experimental exposure via an acute bolus is unlikely to resemble natural infections. Thus, the objectives were to characterize effects of chronic endotoxemia on production parameters and follicular development in estrous-synchronized lactating cows. Eleven Holstein cows (169 ± 20 d in milk; 681 ± 16 kg of body weight) were acclimated to their environmental surroundings for 3 d and then enrolled in 2 experimental periods (P). During P1 (3 d) cows consumed feed ad libitum and baseline samples were obtained. During P2 (7 d), cows were assigned to continuous infusion of either (1) saline-infused and pair-fed (CON-PF; 40 mL/h of saline i.v.; n = 5) or (2) LPS infused and ad libitum fed (LPS-AL; Escherichia coli O55:B5; 0.017, 0.020, 0.026, 0.036, 0.055, 0.088, and 0.148 µg/kg of body weight/h i.v. on d 1 to 7, respectively; n = 6). Controls were pair-fed to the LPS-AL group to eliminate confounding effects of dissimilar nutrient intake. Infusing LPS temporally caused mild hyperthermia on d 1 to 3 (+0.49°C) relative to baseline. Dry matter intake of LPS-AL cows decreased (28%) on d 1 of P2, then progressively returned to baseline. Relative to baseline, milk yield from LPS-AL cows was decreased on d 1 of P2 (12%). No treatment differences were observed in milk yield during P2. Follicular growth, dominant follicle size, serum progesterone (P4), and follicular P4 and 17ß-estradiol concentrations were similar between treatments. Serum 17ß-estradiol tended to increase (115%) and serum amyloid A and LPS-binding protein were increased (118 and 40%, respectively) in LPS-AL relative to CON-PF cows. Compared with CON-PF, neutrophils in LPS-AL cows were initially increased (45%), then gradually decreased. In contrast, monocytes were initially decreased (40%) and progressively increased with time in the LPS-AL cows. Hepatic mRNA abundance of cytochrome P450 family 2 subfamily C (CYP2C) or CYP3A was not affected by LPS, nor was there a treatment effect on toll-like receptor 4 or LBP; however, acyloxyacyl hydrolase and RELA subunit of nuclear factor kappa B tended to be increased in LPS-AL cows. These data suggest lactating dairy cows become tolerant to chronic and exponentially increasing LPS infusion in terms of production and reproductive parameters.

Feeding distillers dried grains in replacement of forage in limit-fed dairy heifer rations: Effects on metabolic profile and onset of puberty.

The objective of this study was to determine the effect of increasing the inclusion rate of distillers dried grains (DDGS) in replacement of forage in limit-fed diets on the metabolic profile and onset of puberty in dairy heifers. A 16-wk randomized complete block design study was conducted using 48 Holstein heifers (199 ± 2 d of age) with 3 treatments. Treatments were (1) 30% DDGS (30DG), (2) 40% DDGS (40DG), and (3) 50% DDGS (50DG), with the remainder of the diet consisting of grass hay and 1.5% mineral mix. Heifers were housed in groups of 6 and individually limit-fed using Calan gates at 2.65, 2.50, and 2.35% of body weight (BW) on a dry matter (DM) basis for 30DG, 40DG, and 50DG, respectively. Jugular blood samples were collected during wk 0, 4, 8, 12, and 16 for metabolite and metabolic hormone analysis. Additional samples were taken during wk 16 for plasma fatty acid analysis. When heifers weighed 200 kg, coccygeal vein blood samples were taken twice per week for progesterone analysis to estimate onset of puberty. Blood samples continued until cycling was confirmed via ultrasound for the presence of a corpus luteum. A quadratic response and a linear tendency in the proportion of total fatty acids as linoleic acid were observed. Linear and quadratic responses for plasma concentrations of linoleic acid and arachidonic acid occurred. Overall results for fatty acid analysis demonstrated that total fatty acid and polyunsaturated fatty acids concentration in the blood were linearly increased, with a quadratic response for polyunsaturated fatty acids with 30DG and 50DG having the greatest concentrations. No interactions of treatment by week were observed for any of the metabolites and metabolic hormones measured. Glucose, insulin, insulin-like growth factor-1, leptin, and triglycerides were similar across treatments. A linear response of plasma urea nitrogen and a quadratic response tendency for cholesterol concentration were observed. Age and BW at puberty were similar across treatments. Limit-feeding heifers with greater inclusion rates of DDGS maintained energy status without the accumulation of excess adipose tissue as indicated by leptin. Treatments had no detrimental effects on age or BW at puberty.

Feeding distillers dried grains in replacement of forage in limit-fed dairy heifer rations: Effects on growth performance, rumen fermentation, and total-tract digestibility of nutrients.

The objective of this study was to determine the effects of increasing dietary concentration of distillers dried grains (DDGS) in dairy heifer rations. A 16-wk randomized complete block design study was conducted using 48 Holstein heifers [199±2 d of age; body weight (BW) 206±2kg] to evaluate effects of dietary treatment on dry matter (DM) intake, average daily gain, growth performance, rumen fermentation, and nutrient digestibility. Treatments were (1) 30% DDGS with the diet fed at 2.65% of BW, (2) 40% DDGS with the diet fed at 2.50% of BW, and (3) 50% DDGS with the diet fed at 2.35% of BW. The remainder of the diet consisted of grass hay and 1.5% mineral mix. Heifers were individually limit-fed using Calan gates. Heifers were weighed every 2 wk and the ration amount offered was adjusted accordingly. Frame measurements and body condition score were recorded every 2wk. Rumen fluid was collected via esophageal tubing during wk 12 and 16 for pH, ammonia N, and volatile fatty acid analysis. Total-tract digestibility of nutrients was evaluated during wk 16 using fecal grab sampling. No treatment by week interactions were found for any of the growth parameters measured, and growth parameters did not differ among treatments. Heifer DM intake linearly decreased with increasing concentrations of DDGS. Body weight and average daily gain were similar among treatments, whereas gain:feed linearly increased across treatments, with a tendency for a treatment by time interaction. As the dietary concentrations of DDGS increased, rumen ammonia N linearly increased. Acetate proportion and acetate:propionate linearly decreased as DDGS increased, whereas propionate linearly increased. There were treatment by time interactions for propionate proportion and acetate:propionate. Increasing dietary concentrations of DDGS linearly increased total-tract digestibility of DM, organic matter, and crude protein. Limit-feeding diets with greater concentrations of DDGS improved gain:feed and total-tract digestibility of DM and crude protein, and maintained frame growth without increasing body condition score. Results demonstrated that replacing forage with DDGS at up to a 50% inclusion rate in limit-fed rations can maintain heifer growth performance.

Feeding fat from distillers dried grains with solubles to dairy heifers: II. Effects on metabolic profile.

The objective of this study was to determine if increased dietary fat from dried distillers grains with solubles (DDGS) in diets of growing heifers affected metabolic profile, plasma fatty acid profile, and reproductive maturation. Thirty-three Holstein heifers (133±18 d of age) were used in a 24-wk randomized complete block design with 3 treatment diets. Treatment diets were (1) control (CON) containing ground corn (15.9% of DM) and soybean products (17.9%), (2) low-fat (LFDG) containing low-fat DDGS (21.9%) and ground corn (11.9%), or (3) high-fat (HFDG) with traditional DDGS (33.8%). Diets were isonitrogenous and isocaloric, but the HFDG diet was formulated to contain 4.8% fat compared with 2.8% in the CON and LFDG diets. All 3 diets were limit-fed to 2.45% of body weight on a dry matter basis, and resulted in a mean average daily gain of 0.96kg/d across treatments. Every 4wk, jugular blood was collected for analysis of metabolites and metabolic hormones. During wk20 of the feeding period, blood samples were collected for analysis of plasma fatty acid profiles. When heifers weighed between 200 and 300kg of body weight, coccygeal blood samples were taken twice weekly for analysis of progesterone to determine if puberty had been reached. Plasma concentrations of nonesterified fatty acids were similar among treatments and consistent over the duration of the study. Plasma concentrations of ß-hydroxybutyrate, insulin, insulin-like growth factor-1, and leptin were similar among heifers fed each treatment diet, but increased over the duration of the feeding period. Serum concentrations of glucose tended to be less in heifers fed HFDG compared with heifers fed the CON diet. Glucose concentrations fluctuated throughout the feeding period, but no treatment by time interactions were noted. Plasma urea N concentrations were less in heifers fed LFDG compared with heifers fed HFDG and CON diets. The concentrations of plasma urea N increased over the duration of the feeding period, with no treatment by week interaction. Total plasma cholesterol was greater in heifers fed HFDG compared with the CON and LFDG diets, and a significant week effect and a week by treatment interaction were observed. Fatty acid profiles also differed among treatments based on the supply of fatty acids from the diet. Progesterone analysis indicated that heifers fed HFDG tended to be pubertal at a younger age than heifers on CON. These results demonstrate that dietary fat from DDGS can be used in high-plane of nutrition rations for growing heifers and maintain metabolic energy status compared with starch from corn, but alters the concentrations of different blood lipids.

Effect of heifer frame score on growth, fertility, and economics.

A non-traditional forage-based protocol was employed to evaluate replacement heifer growth, fertility, and economics between small frame (SF, 3.50; n = 50) and large frame (LF, 5.56; n = 50) heifers using three increasing gain growth phases. Preceding an 85 d growing-breeding period (Phase 3; P3) the heifers were managed as a common group for Phases 1 and 2 (P1 and P2). During P1, heifers grazed common fields of unharvested corn and corn residue (total digestible nutrients [TDN] 56%) with supplemental hay. For P2, heifers grazed early spring crested wheatgrass pasture (CWG; TDN 62%) that was followed by the final P3 drylot growing and breeding period (TDN 68%). Small frame heifers were lighter at the end of P1 in May and at the start of P3 breeding in August (p = 0.0002). Percent of mature body weight (BW) at the end of P1 (209 d) was 48.7% and 46.8%, respectively, for the SF and LF heifers and the percent pubertal was lower for SF than for LF heifers (18.0% vs 40.0%; p = 0.02). At breeding initiation (P3), the percentage of mature BW was 57.8 and 57.2 and the percentage pubertal was 90.0 and 96.0 (p = 0.07) for the SF and LF heifers, respectively; a 5-fold increase for SF heifers. Breeding cycle pregnancy on days 21, 42, and 63, and total percent pregnant did not differ (p>0.10). In drylot, SF heifer dry matter intake (DMI) was 20.1% less (p = 0.001) and feed cost/d was 20.3% lower (p = 0.001), but feed cost/kg of gain did not differ between SF and LF heifers (p = 0.41). Economically important live animal measurements for muscling were measured in May and at the end of the study in October. SF heifers had greater L. dorsi muscle area per unit of BW than LF heifers (p = 0.03). Small frame heifer value was lower at weaning (p = 0.005) and the non-pregnant ending heifer value was lower for SF heifers than for the LF heifers (p = 0.005). However, the total development cost was lower for SF heifers (p = 0.001) and the net cost per pregnant heifer, after accounting for the sale of non-pregnant heifers, was lower for SF heifers (p = 0.004). These data suggest that high breeding efficiency can be attained among March-April born SF and LF virgin heifers when transitioned to a more favorable May-June calving period through the strategic use of grazed and harvested forages resulting in a lower net cost per pregnant SF heifer.

Influence of commercial inactivated or modified-live virus vaccination at time of AI on corpus luteum development and function in beef cattle.

Our objective was to evaluate the effect of vaccination with an inactivated virus vaccine (IVV) or modified-live virus (MLV) vaccine on the corpus luteum (CL). On d0, synchronized beef cows were treated with MLV (n = 70; BoviShield Gold FP5VL5), IVV (n = 16; ViraShield 6VL5HB), or were unvaccinated controls (n = 5). Plasma was collected from treated animals on d0 and every other day through d22. Plasma was analyzed for concentrations of progesterone and 15 cytokines. Between d10 and d13, selected females (n = 13) were ovariectomized; controls were slaughtered on d15/16 to obtain CL for histological evaluation. There were reduced numbers of large luteal cells (LLC) in MLV compared to IVV and controls (P < 0.0001), but IVV were similar to controls (P = 0.11). MLV had decreased LLC percentage compared to controls, and IVV were intermediate (P < 0.0001, MLV: 1.57 ± 0.33 %, IVV: 2.99 ± 0.30 %, Control: 6.45 ± 0.33 %). Based on progesterone concentrations, 24 % MLV and 0 % IVV had an abnormal cycle following vaccination. Overall, MLV had reduced progesterone concentrations (P = 0.02; MLV: 3.61 ± 0.22; IVV: 4.81 ± 0.46 ng/mL). The new CL that formed following an abnormal cycle in MLV had the greatest percentage (35.56 ± 5.5 %) of apoptotic cells. Treatment by cycle status interaction, and time significantly affected IFN-γ, IP-10, MIP-1ß, and MCP-1 (P < 0.03), with several time points having elevated concentrations in abnormally cycling MLV animals. Collectively, this demonstrates MLV vaccination around estrus negatively influenced LLC, progesterone, and increased luteal apoptosis and pro-inflammatory cytokines.

Timing and duration of nutrient restriction and its impacts on placental development and umbilical blood flow in adolescent sheep.

We hypothesized that nutrient restriction from day 50-90 of gestation decreases umbilical blood flow and that umbilical blood flow would recover to control values upon realimentation during late gestation (d 90 to 130) or remain reduced in ewes that continued to be nutrient restricted. On d 50 of gestation, young nulliparous whiteface ewes (6-8 mo; n = 41) carrying singletons were randomly assigned to two dietary treatments: 100% of NRC recommendations (CON) or 60% of CON (RES). On d 90 of gestation, ewes either remained on CON or RES until d 130, or CON ewes were RES from d 90 to 130, or RES ewes were realimented to CON from d 90 to 130. This resulted in 4 treatment groups on day 130: CON-CON, CON-RES, RES-RES, RES-CON. Umbilical blood flow and fetal and placental measurements were obtained via ultrasonography every 10 days from day 50-110. Non-survival surgeries were performed on days 50, 90, and 130 (n = 6-7 ewes/group) where uterine artery and umbilical blood flows were measured during surgery via ultrasonography. Conceptus weights were recorded and placentomes collected to determine binucleate cell numbers. The study was conducted as a completely randomized design arrangement with repeated measures. Data were analyzed using the MIXED procedure of SAS. There was a nutritional treatment by day interaction (P < 0.01) with CON ewes having greater umbilical blood flow compared with RES by d 90. Fetal biparietal distance, abdominal width, and kidney area increased (P < 0.05) in CON-RES with all these measurements increasing during late gestation. We partially accept our hypothesis as nutrient restriction during mid gestation decreased umbilical blood flow. However, blood flow did not return to control levels upon realimentation. By d 130, fetal and placental weights were similar between RES-RES and CON-CON. Binucleate cell numbers in the fetal trophoblast were not influenced by nutritional treatments. Our findings suggest that refeeding previously nutrient restricted pregnant adolescent ewes to control levels does not reestablish umbilical blood flow. Adequate placental development during mid gestation could protect the fetus from a decreased umbilical blood flow later in gestation when nutrients were limited by 40%.

Influence of preovulatory estradiol treatment on the maintenance of pregnancy in beef cattle receiving in vivo produced embryos.

This experiment was designed to determine the role of preovulatory estradiol in pregnancy retention after embryo transfer (ET). Cows were synchronized with the 7-d CO-Synch + CIDR® protocol. On d0 (d-2 =CIDR® removal), cows were grouped by estrual status (estrual [Positive Control] and nonestrual), and nonestrual cows were administered Gonadotropin Releasing Hormone (GnRH) and randomly assigned to either no treatment (Negative Control) or Estradiol (0.1 mg estradiol 17-ß IM). All cows received an embryo on d7. Pregnancy status was retrospectively classified on d56, 30, 24, and 19 by either ultrasonography, plasma pregnancy-associated glycoproteins analysis (PAGs), expression of interferon-stimulated genes, plasma progesterone (P4) concentrations, or a combination of the factors. There was no difference in estradiol concentrations on day 0 h 0 (P > 0.16). At day 0 h 2, Estradiol cows (15.7 ± 0.25 pg/mL) had elevated (P < 0.001) estradiol compared with Positive Controls (3.4 ± 0.26 pg/mL) or Negative Controls (4.3 ± 0.25 pg/mL). On d19, pregnancy rates did not differ (P = 0.14) among treatments. On d24, Positive Controls (47%) had greater (P < 0.01) pregnancy rates than Negative Controls (32%); Estradiol cows were intermediate (40%). There was no difference (P = 0.38) in pregnancy rates between Positive Control (41%) and Estradiol (36%) cows on d30, but Negative Control (27%) cows had (P = 0.01) or tended (P = 0.08) to have decreased pregnancy rates, respectively. Thus, preovulatory estradiol may elicit an effect on early uterine attachment or alter histotroph components, consequently improving pregnancy maintenance through d30.

RNA sequencing and iTRAQ proteomic data from an experiment examining the influence of conceptus presence and preovulatory estradiol on endometrial gene transcripts and proteins around maternal recognition of pregnancy in beef cattle.

RNA sequencing reads and isobaric tags for a relative and absolute quantification (iTRAQ)-Based Proteomic Data were used to determine the impact of conceptus presence and preovulatory estradiol concentration on function of the d16 uterus in beef cattle. Conceptuses and endometrial biopsies were collected from the uterine horn ipsilateral to the corpus luteum. Total cellular RNA was extracted from endometrium for RNA sequencing across two lanes of a NovaSeq S2, 2 × 50-bp run. Two independent uterine luminal fluid pools (ULF) were made for each group: highE2/conceptus, highE2/noconceptus, lowE2/conceptus, and lowE2/noconceptus. Peptides were labeled with iTRAQ reagents and analyzed using 2-dimensional liquid chromatography mass spectrometry. Transcript abundances were determined using DESeq2 (FDR <0.05, FC>2). Scaffold Q+ was used to quantitate peptide and protein identifications in ULF. Datasets include uterine transcript and protein abundances among highE2/conceptus vs highE2/noconceptus and lowE2/conceptus vs lowE2/noconceptus groups. This information can be useful for further investigating the role of specific transcripts and proteins in the maintenance of early pregnancy in beef cattle. This dataset is related to the article 'Influence of conceptus presence and preovulatory estradiol exposure on uterine gene transcripts and proteins around maternal recognition of pregnancy in beef cattle' by E.J. Northrop-Albrecht, J.J.J. Rich, R.A. Cushman, R. Yao, X. Ge, G.A. Perry. Molecular and Cellular Endocrinology.

iTRAQ-Based proteomic dataset for bovine pre-ovulatory plasma and follicular fluid containing high and low Estradiol.

This is isobaric tags for a relative and absolute quantification (iTRAQ)-Based Proteomic Data on bovine plasma (PL) and follicular fluid (FF) containing high and low pre-ovulatory circulating concentration of estradiol (E2). The PL and FF were collected from nine beef cows that were identified to initiate a new follicular wave on day -4 during synchronization. Follicular dynamics and ovulatory response were monitored using transrectal ultrasonography. Blood samples were collected at slaughter and FF was aspirated from dominant follicles (DF; >10 mm). Estradiol concentrations in PL and FF were measured by radioimmunoassays. Plasma and FF were labeled as containing high E2 (PL HE2 and FF HE2) or low E2 (PL LE2 and FF LE2). Abundant proteins (albumin, IgG, IgA, and alpha-1-antitrypsin) were depleted from the four PL and FF samples. Peptides were labeled with iTRAQ reagents and analyzed using 2-dimentional liquid chromatography ESI-based mass spectrometry. Proteins were identified and quantified using SEQUESTTM search engine embedded in Proteome Discoverer. The proteins matched with at least one unique peptide at minimum 95% confidence were considered positive identifications. Protein expression levels were determined by assigned fold change of >2.0 or <0.5 between any pair from the four sample types. The paired comparisons made were PL HE2 and PL LE2, FF HE2 and FF LE2, PL HE2 and FF HE2, and PL LE2 and FF LE2. Protein Analysis Through Evolutionary Relationships (PANTHER) and Database for Annotation, Visualization and Integrated Discovery (DAVID) were used to classify protein functions. This dataset includes the overview of workflow for identification and quantification of proteins and details on 231 proteins identified which includes 103 up- and down-regulate proteins. This dataset can be useful for further probing of the identified regulated proteins to better understand folliculogenesis and ovulation, particularly in bovine. This dataset is related to the article 'iTRAQ-Based Proteomic Analysis of Bovine Pre-ovulatory Plasma and Follicular Fluid' by P. A. Afedi, E. L. Larimore, R. A. Cushman, D. Raynie, G. A. Perry. Domestic Animal Endocrinology. https://doi.org/10.1016/j.domaniend.2021.106606.

iTRAQ-based proteomic analysis of bovine pre-ovulatory plasma and follicular fluid.

Bovine follicular fluid (FF) creates a unique microenvironment in follicles necessary for follicle growth, oocyte maturation, and estradiol (E2) production. The objective of this study was to analyze changes in proteins in FF and plasma (PL) from animals with high E2 (HE2) or low E2 (LE2) during the preovulatory period. Beef cows were synchronized, and follicular dynamics and ovulatory response were monitored using transrectal ultrasonography. Nine cows were selected and slaughtered, blood samples were collected at slaughter and FF was aspirated from dominant follicles (DF; >10 mm). Abundant proteins (albumin, IgG, IgA, and alpha-1-antitrypsin) were depleted from both PL and FF. Peptides were labeled with iTRAQ reagents and quantified using 2-dimentional liquid chromatography ESI-based mass spectrometry. Estradiol was associated with protein changes in PL and FF. Protein expression changes between FF HE2 and FF LE2 were greater than between PL HE2 and PL LE2. There were 15 up-regulated proteins and 10 down-regulated proteins in FF HE2 compared to FF LE2, and 7 proteins up-regulated and 9 proteins down-regulated in PL HE2 compared to PL LE2. Several of the differentially expressed proteins function in follicle development and were mainly categorized under cellular process and metabolic process. Pathway analysis identified the up- and down-regulated proteins were predominantly associated with the complement and coagulation cascades. The data demonstrate E2 regulates a wide range of reproductive associated proteins in bovine PL and FF and can provide the basis for further investigation of specific processes involved in such regulation.

Acute effects of estradiol-17ß on plasma volume and uterine cell proliferation in sheep.

Ovarian steroids play an important role in increasing plasma volume in pregnant females and preparing the uterus for implantation. We hypothesized that a short duration of increased estradiol-17ß (E2) would increase plasma volume and uterine cell proliferation in ovariectomized ewes. Adult non-pregnant Romanov ewes (n = 15) were ovariectomized. After ovariectomy, ewes were individually housed and were offered water at ad libitum intake and were fed a pelleted diet at maintenance once daily according to body weight. After at least 30 days post-ovariectomy ewes were fasted and received an implant placed in the axillary region that contained 100 mg of E2 (E2; n = 8) or a sham implant with no E2 (CON, n = 7). After 24 h, ewes were weighed prior to plasma volume measurement procedures. Plasma volume was determined using the Evans blue dye method. Blood samples were taken at 0 (pre dye injection), 5, 10, 15, 20, 25, 30, 40, 50 and 60 min after dye injection. After the final blood collection, ewes were euthanized with an overdose of sodium pentabarbital and uterine weights were recorded. Uterine cross-sections were fixed in formalin for immunohistochemical localization of Ki67 (a marker of proliferating cells) followed by image generation of luminal epithelium and endometrial stroma (5 areas each/tissue section) and analysis to determine the proportion of proliferating cells. Plasma volume tended to be greater in E2 vs CON (2.75 ± 0.11 vs. 2.54 ± 0.12 L, P = 0.07) and uterine weights were greater in E2 vs CON (27.25 ± 2.35 vs. 17.35 ± 2.51 g, P < 0.01). Water intake after implant placement was similar in E2 and CON (3.85 vs. 4.87 ± 0.67 L; P = 0.28). Cell proliferation in the luminal epithelium was greater in E2 vs CON (6.55 vs. 1.2 ± 1.75%, P = 0.02) and stromal cells tended to be greater in E2 vs CON (0.59 vs 0.37 ± 0.06%, P = 0.07). Our results demonstrate that E2-treatment tends to increase plasma volume acutely and increases uterine cell proliferation in ewes.

Effect of estradiol preceding and progesterone subsequent to ovulation on proportion of postpartum beef cows pregnant.

Two experiments were conducted to examine the effect of plasma concentrations of 17ß-estradiol (E2) preceding and progesterone (P4) subsequent to ovulation on proportions of beef cows pregnant following embryo transfer. Timing of ovulation (d 0) among postpartum cows was synchronized and cows that expressed estrus were removed from each study. In Experiment 1, plasma E2 concentration on d 0 was used to classify cows (n = 353) into Low, Medium, and High E2 groups. Pregnancy rate for cows with Low, Medium, or High E2 concentrations were different (P < 0.05). In Experiment 2, there were multiple administrations of PGF2α to evaluate the independent effects of Low or High E2 before ovulation and Low or Normal (no treatment) P4 after ovulation on proportions of cows pregnant. Treatment groups in Experiment 2, therefore, were: Low E2-Low P4 (LL; n = 71), Low E2-Normal P4 (LN; n = 69), High E2-Low P4 (HL; n = 74), and High E2-Normal P4 (HN; n = 73). Concentrations of P4 on d 7 subsequent to ovulation were less (P < 0.05) in cows of the HL compared to HN, and in LL compared to LN groups. Concentrations of E2 on d -2, 0, and change in E2 (d -2 to d 0) had a positive effect (P < 0.008) on pregnancy rates. In summary, relatively greater E2 concentrations preceding ovulation; and relatively greater P4 concentrations subsequent to ovulation combined with lesser E2 concentrations preceding ovulation had a positive effect on proportions of postpartum cows pregnant.

Pre-synchronization of ovulation timing and delayed fixed-time artificial insemination increases pregnancy rates when sex-sorted semen is used for insemination of heifers.

This study was conducted to determine effects of pre-synchronization of ovulation timing among heifers and delayed fixed-time artificial insemination (TAI) with sex-sorted semen on proportion of heifers pregnant after TAI (PR/AI). Heifers were assigned to one of eight treatments: 1 and 2), 7-d CO-Synch + CIDR treatment regimen with administration of gonadotropin-releasing hormone and a CIDR insert on Day 0, prostaglandin F2α (PGF) at CIDR removal on Day 7, and TAI occurring 54 h later with conventionally processed (CTRL54-CNV) or sex-sorted semen (CTRL54-SEX); 3 and 4), same as CTRL54 but TAI delayed to 72 h with conventionally processed (CTRL72-CNV) or sex-sorted semen (CTRL72-SEX); 5 and 6), same as CTRL54 but additional administration of PGF on Day -7 and TAI with conventionally processed (PRE54-CNV) or sex-sorted semen (PRE54-SEX); 7 and 8), same as PRE54 treatments but TAI delayed to 72 h with conventionally processed (PRE72-CNV) or sex-sorted semen (PRE72-SEX). Proportion of heifers pregnant after TAI was greater (P ≤  0.02) with conventionally processed semen compared with sex-sorted semen, yet PR/AI did not differ (P =  0.14) between heifers in PRE72-CNV and PRE72-SEX groups. There were greater PR/AI in the PRE72-SEX (P =  0.03) than CTRL54-SEX group (46.1 % and 36.9 %) and there was no difference (P =  0.31) in PR/AI between CTRL54-CNV and PRE72-SEX groups (50.4 % and 46.1 %). In conclusion, pre-synchronization of ovulation timing among heifers combined with delayed TAI resulted in increased PR/AI with sex-sorted semen compared with the 7-d CO-Synch+CIDR treatment regimen.

GnRH treatment at artificial insemination in beef cattle fails to increase plasma progesterone concentrations or pregnancy rates.

Treatment with GnRH at the onset of standing estrus increased pregnancy percentages and circulating concentrations of progesterone in repeat breeder dairy cows. The objective of this study was to determine the effect of treatment with GnRH at AI on concentrations of progesterone and conception rates in beef cattle that exhibited estrus. Two hundred ninety-three heifers at four locations were synchronized with the Select Synch plus CIDR protocol (given GnRH and a CIDR was placed into the vagina, and 7 d later, given PGF(2alpha) and CIDR removed; n=253) or the 14-19 melengestrol acetate (MGA) protocol (MGA fed at 0.5mg/head/d for 14 d, with PGF(2alpha) 19 d after MGA withdrawal n=40) and AI was done after detection of estrus. At Location 1, blood samples were collected on Day 2, 4, 6, 10, 15, and 18 after AI (Day 0=AI). Two hundred and fifty postpartum cows at two locations were synchronized with the Select Synch plus CIDR protocol, and AI was performed after detection of estrus. At AI, cattle were alternately assigned to one of two treatments: (1) treatment with GnRH (100microg) at AI (n=127 heifers and n=108 cows); or (2) non-treated control (n=120 heifers and n=119 cows). Concentrations of progesterone tended to be greater in control heifers compared to GnRH-treated heifers on Days 6 (P=0.08), 10 (P=0.07), and 15 (P=0.11). Overall conception rates were 68% and 66% for GnRH treated and control, respectively, and were not different between treatments (P=0.72). In summary, treatment with GnRH at time of AI had no influence on conception rates in cattle that had exhibited estrus.

Effect of preovulatory concentrations of estradiol and initiation of standing estrus on uterine pH in beef cows.

Previous studies have indicated that initiation of standing estrus within 24h of fixed-time AI influenced pregnancy rates. Furthermore, uterine environment at time of insemination can influence sperm transport. We hypothesized that preovulatory concentrations of estradiol would influence uterine pH at time of insemination. The objective of this study was to determine the influence of elevated preovulatory concentrations of estradiol on uterine pH following a fixed-time AI protocol. Cows were synchronized with the CO-Synch (n=57) protocol, and 29 cows were treated with an injection of estradiol cypionate (ECP; 1mg) 36h before the second injection of GnRH. Cows that exhibited standing estrus or were treated with ECP had increased (P<0.05) concentrations of estradiol compared to cows not in estrus and not administered ECP, respectively. There was an ECP by standing estrus interaction on uterine pH (P=0.01). Control cows that exhibited estrus had a reduced uterine pH (6.72+/-0.10; P=0.05) compared to control cows not exhibiting estrus (7.0+/-0.06). Cows treated with ECP and detected in standing estrus had a greater uterine pH (7.0+/-0.07) compared to control cows in estrus (P=0.02) and ECP cows not in estrus (6.81+/-0.09; P=0.06). The interval between the initiation of standing estrus and when pH was determined also influenced uterine pH. Cows that initiated standing estrus within 4h of pH determination had a lower uterine pH (6.74+/-0.12) compared to cows that initiated estrus 4-8h (7.09+/-0.08; P=0.07) or 8-12h (7.10+/-0.15; P=0.03) after pH determination. In summary, elevated concentrations of estradiol influenced standing estrus but only influenced uterine pH when pH was determined within 4h of the initiation of standing estrus.

Effects of excess metabolizable protein on ovarian function and circulating amino acids of beef cows: 1. Excessive supply from corn gluten meal or soybean meal.

In the dairy industry, excess dietary CP is consistently correlated with decreased conception rates. However, the source from which excess CP is derived and how it affects reproductive function in beef cattle is largely undefined. The objective of this experiment was to determine the effects of feeding excess metabolizable protein (MP) from feedstuffs differing in rumen degradability on ovulatory follicular dynamics, subsequent corpus luteum (CL) development, steroid hormone production and circulating amino acids (AA) in beef cows. Non-pregnant, non-lactating mature beef cows (n=18) were assigned to 1 of 2 isonitrogenous diets (150% of MP requirements) designed to maintain similar BW and body condition score (BCS) between treatments. Diets consisted of ad libitum corn stalks supplemented with corn gluten meal (moderate rumen undegradable protein (RUP); CGM) or soybean meal (low RUP; SBM). After a 20-day supplement adaptation period, cows were synchronized for ovulation. After 10 days of synchronization, gonadotropin releasing hormone (GnRH) was administered to reset ovarian follicular growth. Starting at GnRH administration and daily thereafter until spontaneous ovulation, transrectal ultrasonography was used to diagram ovarian follicular growth, and blood samples were collected for hormone, metabolite and AA analyses. After 7 days of visual detection of estrus, CL size was determined via ultrasound. Data were analyzed using the MIXED procedures of SAS. As designed, cow BW and BCS were not different (P⩾0.33). Ovulatory follicular wavelength, antral follicle count, ovulatory follicle size at dominance and duration of dominance were not different (P>0.13) between treatments. Cows supplemented with CGM had greater post-dominance ovulatory follicle growth, larger dominant follicles at spontaneous luteolysis, shorter proestrus, and larger ovulatory follicles (P⩽0.03) than SBM cows. No differences (P⩾0.44) in peak estradiol, ratio of estradiol to ovulatory follicle volume, or plasma urea nitrogen were observed. While CL volume and the ratio of progesterone to CL volume were not affected by treatment (P⩾0.24), CGM treated cows tended to have decreased (P=0.07) circulating progesterone 7 days post-estrus compared with SBM cows. Although total circulating plasma AA concentration did not differ (P=0.70) between treatments, CGM cows had greater phenylalanine (P=0.03) and tended to have greater leucine concentrations (P=0.07) than SBM cows. In summary, these data illustrate that excess MP when supplemented to cows consuming a low quality forage may differentially impact ovarian function depending on ruminal degradability of the protein source.

Effects of excess metabolizable protein on ovarian function and circulating amino acids of beef cows: 2. Excessive supply in varying concentrations from corn gluten meal.

In the dairy industry, excess dietary CP is consistently correlated with decreased conception rates. However, amount of excess CP effects on reproductive function in beef cattle is largely undefined. The objective of this experiment was to determine the effects of excess metabolizable protein (MP) supplementation from a moderately abundant rumen undegradable protein (RUP) source (corn gluten meal: 62% RUP) on ovarian function and circulating amino acid (AA) concentrations in beef cows consuming low quality forage. Non-pregnant, non-lactating beef cows (n=16) were allocated by age, BW and body condition score (BCS) to 1 of 2 isocaloric supplements designed to maintain BW for 60 days. Cows had ad libitum access to corn stalks and were individually offered a corn gluten meal-based supplement daily at 125% (MP125) or 150% (MP150) of National Research Council (NRC) MP requirements. After a 20-day supplement adaptation period, cows were synchronized for ovulation. After 10 days of synchronization, follicular growth was reset with gonadotropin releasing hormone. Daily thereafter, transrectal ultrasonography was performed to diagram ovarian follicular waves, and blood samples were collected for hormone, metabolite and AA analyses. After 7 days of observation of estrus, corpus luteum (CL) size was determined via ultrasound. Data were analyzed using the MIXED procedures of SAS. No differences (P⩾0.21) in BW and BCS existed throughout the study; however, plasma urea N at ovulation was greater (P=0.04) in MP150. Preovulatory ovarian follicle size at dominance, duration of dominance, size at spontaneous luteolysis, length of proestrus and wavelength were not different (P⩾0.11) between treatments. However, ovulatory follicles were larger (P=0.04) and average antral follicle count was greater (P=0.01) in MP150 than MP125. Estradiol concentration and ratio of estradiol to ovulatory follicle volume were not different due to treatment (P⩾0.25). While CL volume 7 days post-estrus was greater (P<0.01) in MP150 than MP125, circulating progesterone 7 days post-estrus and ratio of progesterone to CL volume were not different (P⩾0.21). Total AA were not different (P⩾0.76) at study initiation or completion; however, as a percent of total AA, branched-chain AA at ovulation were greater (P=0.02) in MP150. In conclusion, supplementation of CP at 150% of NRC MP requirements from a moderately undegradable protein source may enhance growth of the ovulatory follicle and subsequent CL compared with MP supplementation at 125% of NRC MP requirements.

Effects of supplementing calcium salts of polyunsaturated fatty acids to late-gestating beef cows on performance and physiological responses of the offspring.

This experiment compared performance and physiological responses of the offspring from cows supplemented with Ca salts of PUFA or SFA + MUFA during late gestation. Ninety-six multiparous, nonlactating, pregnant Angus × Hereford cows were ranked by BW, BCS, and age and divided into 24 groups of 4 cows/group at the end of their second trimester of gestation (d -7). Cows conceived during the same estrus synchronization + AI protocol, with semen from a single sire; hence, gestation length was 195 d for all cows at the beginning of the experiment (d 0). Groups were randomly assigned to receive (DM basis) 405 g/cow daily of soybean meal in addition to 1) 190 g/cow daily of Ca salts of PUFA based on eicosapentaenoic, docosahexaenoic, and linoleic acids or 2) 190 g/cow daily of Ca salts of SFA + MUFA based on palmitic and oleic acids (CON). Groups were maintained in 2 pastures (6 groups of each treatment/pasture) and received daily 10.1 kg/cow (DM basis) of grass-alfalfa hay. Groups were segregated into 1 of 12 drylot pens (6 by 18 m) and individually offered treatments 3 times/wk from d 0 until calving. Cow BW and BCS were recorded, and blood samples were collected on d -7 of the experiment and also within 12 h after calving. Calf BW was also recorded within 12 h of calving. Calves were weaned on d 280 of the experiment, preconditioned for 45 d (d 280 to 325), transferred to a growing lot on d 325, and moved to a finishing lot on d 445, where they remained until slaughter. At calving, PUFA-supplemented cows had a greater ( < 0.01) proportion (as % of total plasma fatty acids) of PUFA, including linoleic, linolenic, arachidonic, docosapentaenoic, and docosahexaenoic acids. At weaning, calves from CON-supplemented cows were older ( = 0.03), although no treatment differences were detected ( = 0.82) for calf weaning BW. During both growing and finishing phases, ADG was greater ( ≤ 0.06) in calves from PUFA-supplemented cows. Upon slaughter, HCW and marbling were also greater ( ≤ 0.05) in calves from PUFA-supplemented cows. Collectively, these results indicate that supplementing eicosapentaenoic, docosahexaenoic, and linoleic acids to late-gestating beef cows stimulated programming effects on postnatal offspring growth and carcass quality. Therefore, supplementing late-gestating beef cows with Ca salts of PUFA appears to optimize offspring productivity in beef production systems.

Supplementation of metabolizable protein during late gestation and fetal number impact ewe organ mass, maternal serum hormone and metabolite concentrations, and conceptus measurements.

To examine the effects of maternal metabolizable protein (MP) supplementation during late gestation on serum hormone and metabolites and organ masses, multiparous ewes (n = 45) carrying singletons or twins were allotted randomly (within pregnancy group) to 1 of 3 treatments: 60% (MP60), 80% (MP80), or 100% (MP100) of MP requirements. Blood samples were drawn before the initiation of diets (day 100) and before slaughter (day 130) for chemistry panel analysis and weekly for hormone analysis including progesterone (P4) and estradiol-17ß (E2). At day 130, ewe organ masses were recorded. Despite being fed isocaloric diets, MP60 ewes gained less weight throughout pregnancy compared with MP80 and MP100 ewes which were similar. Although diet did not impact E2 or P4 concentrations, ewes carrying twins had greater (P < 0.05) concentrations of both as gestation advanced. Albumin, aspartate aminotransferase, and total protein were reduced (P < 0.05) in MP60 compared with MP100 ewes near term. There was a diet by fetal number interaction (P = 0.03) for lactate dehydrogenase. Twin-carrying MP80 ewes had greater lactate dehydrogenase compared with all other groups on day 130 of gestation. Ewes that were fed MP80 had greater body weight on day 130 of gestation compared with MP60 ewes. Kidney and heart weights were lighter in MP60 ewes compared with MP80 ewes. There was a maternal diet by fetal number interaction (P = 0.05) on fetal weight per unit empty ewe body weight. In ewes carrying singletons, MP60 ewes supported less fetal weight compared with MP100. In contrast, MP60 ewes supported more fetal mass compared with MP100 ewes when carrying twins. The level of protein, and not just total energy, in the diet appears to impact some aspects of the maternal system. Moreover, it appears some measurements of mobilizing maternal body resources are enhanced in ewes carrying twins.