V. R. G. Mercadante

Synchronization and Artificial Insemination Strategies in Beef Cattle

Utilization of estrus or ovulation synchronization and fixed-timed artificial insemination (TAI) has facilitated the widespread utilization of artificial insemination (AI) and can greatly impact the economic viability of cow-calf systems by enhancing weaning weights. Implementation of TAI programs by beef producers results in limited frequency of handling cattle and elimination of the need to detect estrus. Continued use of intensive reproductive management tools such as estrus synchronization and AI will result positive changes to calving distribution, pregnancy rates, and subsequent calf value.

Effects of recombinant bovine somatotropin administration at breeding on cow, conceptus, and subsequent offspring performance of beef cattle.

The effects of administration of recombinant bovine ST (bST) on plasma hormone concentrations of cows, conceptus development, and postnatal calf performance were examined. Lactating beef cows ( = 190) were exposed to a fixed-time AI (TAI) protocol from d -10 to 0 (TAI on d 0). Cows were blocked by breed and stratified by days postpartum and then randomly assigned to receive, subcutaneously 1) 2 injections of saline (1 mL of 0.9% saline), 1 on d 0 at TAI and a second injection on d 14 (CTRL; = 53); 2) an injection of 325 mg of bST on d 0 and a saline injection on d 14 (bST0; = 48); 3) a saline injection on d 0 and an injection of 325 mg of bST on d 14 (bST14; = 49); or 4) 2 injections of 325 mg of bST, 1 on d 0 and a second injection on d 14 (bST0+14; = 40). Pregnancy status, crown-to-rump length (CRL) on Day 35, and crown-to-nose length (CNL) on Day 65 were determined via transrectal ultrasonography. Blood samples were collected on d 0, 7, 14, 21, 35, and 65, relative to TAI, to determine plasma concentrations of progesterone (P4), IGF-1, and pregnancy-specific protein B (PSPB) and also on d 18 and 21 for isolation of peripheral blood leukocytes for RNA extraction and measurement of interferon-stimulated genes transcript abundance. Individual calf BW was determined at birth and every 30 d until weaning. A subset of 24 calves was randomly selected for liver biopsies at birth to determine mRNA expression of target genes. Administration of bST to cows increased ( < 0.0001) concentrations of plasma IGF-1 for 14 d after injection compared with CTRL but did not affect fetal CRL and CNL ( = 0.23). Cows receiving bST only on d 0 had a greater ( = 0.05) transcript abundance in myxovirus resistance 2 on d 21 compared with 2bST cows (2.0- and 0.8-fold for bST0 and 2bST, respectively), whereas cows receiving bST14 and CTRL were intermediate (1.2- and 0.9-fold, respectively). Calf BW did not differ ( ≥ 0.100) among treatments on d 0, 30, 60, 90, 120, and 150 relative to birth. Injection of bST only on d 0 tended ( = 0.062) to increase calf liver mRNA expression of at birth compared with the calves born to cows in other treatments. Therefore, during a TAI protocol, the administration of 1 or 2 injections of 325 mg of bST to lactating beef cows enhanced their plasma concentrations of IGF-1 but failed to improve fetal size and plasma concentrations of maternal PSPB and P4 and had no effect on postnatal calf growth performance.

Effects of molasses and crude glycerol combined in a liquid supplement on ruminal fermentation in beef steers consuming bermudagrass hay

Two experiments were performed to evaluate the effects of 1) increasing supplementation doses of a 50:50 (as-fed) liquid supplement of molasses and crude glycerol (M:G) on ruminal fermentation parameters and blood urea nitrogen (BUN) in beef steers consuming Tifton 85 bermudagrass ( spp.) hay and 2) different proportions of molasses and crude glycerol in a liquid supplement on in vitro fermentation and gas production kinetics. For Exp. 1, 8 ruminally cannulated, Angus-crossbred steers were used in a duplicated 4 × 4 Latin square design, had ad libitum access to Tifton 85 bermudagrass hay, and were randomly assigned to 1 of 4 treatments: 1) CTRL, no supplementation; 2) SUP1, 0.45 kg/d (as fed) of 50:50 M:G; 3) SUP3, 1.36 kg/d (as fed) of 50:50 M:G; and 4) SUP5, 2.27 kg/d (as fed) of a 50:50 M:G. For Exp. 2 in vitro batch cultures were conducted to test the same treatments from Exp. 1 and effects of different proportions of a M:G mixture (100:0, 75:25, 50:50, 25:75, and 0:100) when added to a hay substrate simulating the proportions of hay and liquid supplement used in SUP5. In Exp. 1, increasing doses of liquid supplement linearly decreased ( < 0.001) concentrations of NH-N, BUN, and acetate molar proportions, whereas propionate ( = 0.002) and butyrate ( < 0.001) molar proportions increased linearly. Treatment × time interactions were observed for ruminal pH ( < 0.001), where the greatest decrease was observed at 3 h postfeeding for animals consuming SUP5 (from 6.82 at 0 h to 6.32 at 3 h). In Exp. 2, decreases in acetate molar proportions ( < 0.001) and increases ( < 0.001) in propionate and butyrate molar proportions were also observed for either increasing doses of a 50:50 mixture or increasing proportions of glycerol in the mixture. Total VFA and in vitro organic matter digestibility were increased linearly ( < 0.001) as doses of a 50:50 mixture increased. Increasing doses of 50:50 M:G to growing beef heifers consuming bermudagrass hay caused a shift in VFA profile toward increases in propionate and decreases in acetate molar proportions. This was also confirmed in vitro, as the proportions of crude glycerol increased in a molasses:crude glycerol mix. Thus, molasses and crude glycerol combined seem to be useful to enhance performance in growing cattle consuming forage-based diets.

Supplementing calcium salts of soybean oil after artificial insemination increases pregnancy success in Bos taurus beef cows1

© The Author(s) 2018. Published by Oxford University Press on behalf of the American Society of Animal Science. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com. Transl. Anim. Sci. 2018.2:S9–S13 doi: 10.1093/tas/txy017

Supplementing Ca salts of soybean oil after artificial insemination increases pregnancy success in Bos taurus beef cows1

Two experiments investigated the effects of supplementing Ca salts of soybean oil (CSSO) during early gestation on reproductive function and pregnancy rates to AI in Bos taurus beef cows. In Exp. 1, 771 suckled, lactating, multiparous Angus cows were divided into 22 groups of approximately 35 cows per group and timed inseminated on day 0. After AI, groups were assigned randomly to receive (as-fed basis) 100 g of ground corn + 100 g of soybean meal per cow/d, in addition to 1) 100 g/cow daily of CSSO (n = 11) or 2) 87 g of prilled saturated fat + 13 g of limestone per cow/d (CON; n = 11). Groups were maintained in individual tall fescue-dominated pastures and offered treatments from day 0 to 21. Pregnancy status was determined between days 45 and 55 via transrectal ultrasonography. Cows receiving CSSO had greater (P = 0.01) pregnancy rates to timed AI compared with CON (60.2 vs. 51.7%; SEM = 4.2). In Exp. 2, 90 suckled, lactating, multiparous Angus × Hereford cows housed in 18 drylot pens (5 cows per pen) were assigned to the same timed AI program and treatments from Exp. 1 (9 pens per treatment) and received 20 kg/d (DM basis) of grass-alfalfa hay. Transrectal ultrasonography was performed to verify ovulation and corpus luteum (CL) volume before AI (day 0), on days 7 and 15. After ultrasonography on day 15, cows diagnosed without a CL on day 0, but with a CL greater than 0.38 cm3 in volume on days 7 and 15 (2 or 3 cows per pen; CSSO, n = 20; CON, n = 24), were assigned to conceptus collection via transcervical flushing and endometrial biopsy in the uterine horn ipsilateral to the CL. Blood samples were collected for FA analysis on days 0, 7, and 15. Blood was collected from cows not assigned to conceptus collection for whole-blood RNA extraction on day 20 and for pregnancy diagnosis on day 30 by measuring concentrations of pregnancy-associated glycoproteins. Cows receiving CSSO had greater (P ≤ 0.04) mean plasma concentrations of linoleic acid and ω-6 FA compared with CON on days 7 and 15. Moreover, CSSO supplementation increased (P = 0.05) mRNA expression of interferon-tau by the conceptus and blood mRNA expression of interferon-stimulated gene 15 and 20,50-oligoadenylate synthetase on day 20 in gestating cows. Hence, post-AI CSSO supplementation to B. taurus beef cows improved pregnancy rates to timed AI, which can be associated with increased mRNA expression of interferon-tau by the conceptus when CSSO is supplemented during early gestation.

Administration of recombinant bovine somatotropin prior to fixed-time artificial insemination and the effects on fertility, embryo, and fetal size in beef heifers

Our objectives were to determine the effects of the administration of recombinant bovine somatotropin (bST) at the initiation of a fixed-time AI (TAI) protocol on concentrations of plasma IGF-1, follicle diameter, embryo/fetal size, and pregnancy rates in replacement beef heifers. Four hundred and fourteen Angus-based beef heifers were enrolled in a completely randomized design at 4 locations from January to July of 2016. All heifers were exposed to the 7-d CO-Synch + controlled internal drug release (CIDR) protocol where they received a 100-µg injection of GnRH and a CIDR insert on day -9, 25 mg of PGF2α at CIDR removal on day -2, followed by a 100-µg injection of GnRH and TAI 54 ± 2 h later on day 0. Within location, all heifers were randomly assigned to 1 of 2 treatments: 1) heifers that received 650 mg of bST on day -9 (BST; n = 191); or 2) heifers that did not receive bST on day -9 (CONTROL; n = 223). Blood samples were collected on day -9, 0, 28, and 60 to determine the plasma concentrations of IGF-1. Follicle diameter was determined on day -2 and 0 by transrectal ultrasonography. Pregnancy was diagnosed via transrectal ultrasonography on day 28 or 35, and again at least 30 d after the end of the breeding season. Embryo morphometry was assessed by measuring crown-to-rump length (CRL) on day 28, and fetal size was assessed by measuring crown-to-nose-length (CNL) on day 60. Concentrations of plasma IGF-1 did not differ between treatments on day -9 (P = 0.924), 28 (P = 0.075), and 60 (P = 0.792); however, concentrations of plasma IGF-1 were greater (P < 0.001) in BST-treated heifers at TAI (372.4 ± 16.6 vs. 193.7 ± 16.6 ng/ml). No differences (P = 0.191) were detected for follicle diameter between CONTROL and BST treatments on day -2 or 0. Pregnancy rates to TAI (PR/AI) were greater (P = 0.028) for CONTROL compared to BST heifers (42.5 ± 4.0 vs. 29.9 ± 4.1%). No differences (P = 0.536) in CRL were observed on day 28 between CONTROL and BST heifers. In addition, no difference (P = 0.890) was observed for CNL between CONTROL and BST treatments. Final pregnancy rates did not differ (P = 0.699) between treatments. The administration of bST to beef heifers at the initiation of a TAI protocol increased plasma concentrations of IGF-1 at TAI; however, failed to enhance follicle diameter, embryo/fetal size, and reduced PR/AI.

Nerve Growth Factor-β production in the bull: Gene expression, immunolocalization, seminal plasma constitution, and association with sire conception rates

Nerve Growth Factor-β (NGF) is a seminal plasma protein that regulates sperm physiology in bulls, yet its production and association with fertility remain unclear. The objective of this study was to describe NGF distribution in bull accessory sex glands, quantify its seminal plasma concentrations, and determine its association with sire conception rates. Accessory sex glands were collected post-mortem from mature bulls (n = 3). Quantitative PCR and immunohistochemistry were performed in tissue samples. Pre-ejaculate and sperm-rich fraction of seminal plasma from Angus (n = 42) and sperm-rich fraction from Holstein (n = 10) bulls were collected by electroejaculation. Holstein (n = 76) and Jersey (n = 14) bulls were collected by artificial vagina, and those with available sire conception rate scores were allocated to: (1) negative/0 (n = 15); or (2) positive (n = 45). Seminal plasma NGF concentrations were measured using ELISA. Relative abundance of NGF mRNA was greatest in the vesicular, intermediate in ampulla, and lowest in prostate and bulbourethral glands (P ≤ 0.03). The most intense NGF staining was detected in the ampulla and vesicular glands (P < 0.05). Concentrations of NGF were greater in the sperm-rich fraction than in the pre-ejaculate and in samples collected by artificial vagina (P < 0.01). Seminal plasma NGF concentrations were greater in bulls with positive sire conception rate than those with negative/0 (P = 0.05). This study determined that NGF is produced in the ampulla and vesicular glands of bulls and is secreted into the sperm-rich fraction of the ejaculate. The positive association of NGF with bull fertility warrants further investigation.

13 EFFECTS OF NERVE GROWTH FACTOR-β, PURIFIED FROM BULL SEMINAL PLASMA, ON CORPUS LUTEUM FUNCTION AND CONCEPTUS DEVELOPMENT IN COWS

Nerve growth factor-β (NGF) has been identified in the seminal plasma of livestock and is required for the induction of ovulation in camelids. Until recently, it was thought to play a negligible role in species with spontaneous ovulation. On the contrary, recent studies have shown that systemic administration of NGF, purified from llama seminal plasma, was associated with larger corpus luteum (CL) diameter and higher concentrations of progesterone (P) following ovulation. The objective of the current project was to determine if systemic administration of NGF, purified from bovine seminal plasma, would improve CL formation and enhance embryonic development. Our hypothesis was that systemic administration of NGF at the time of artificial insemination in cows would lead to increased CL volume, increased P secretion, and improved expression of markers of conceptus development and maternal recognition of pregnancy. Seminal plasma was harvested from semen collected by electroejaculation in Angus cross bulls. Purification of NGF was performed using a combination of anion- and cation-exchange chromatography and gradient elution. Beef cows were randomly assigned to CONT (n=30) or NGF (n=30) groups and synchronized using a 7-day Co-Synch+CIDR program. At time of insemination (day 0), NGF cows received 296µg of purified NGF, reconstituted in 12mL of PBS, and CONT cows received 12mL of PBS intramuscularly. Blood samples were collected from the coccygeal vein of each cow at days 0, 3, 7, 10, 14, 19, 21, 28, 31, 38, 45, and 66. Ultrasound was performed at each time point for determination of ovarian structures (day 0), corpus luteum volume (all time points), and fetus detection (day 28). Statistical analysis was performed using analysis of variance with repeated-measures in R (R version 3.2.2; https://www.r-project.org/). At day 28, 17/30 (57%) CONT cows and 21/30 (70%) NGF cows were diagnosed as pregnant (P=0.15). At day 0, cows that later became pregnant had a larger follicular diameter (1.46cm) than those diagnosed as open (1.16cm; P<0.01). Follicular diameter at day 0 did not differ significantly between CONT and NGF groups for either open (P=0.35) or pregnant (P=0.90) cows. CL volume in open cows was affected by day (P<0.001) with no treatment (P=0.84) or treatment by day (P=0.42) interaction. CL volume in pregnant cows was affected by both day (P<0.001) and treatment (P<0.001), with CONT cows having a higher CL volume than NGF cows. Though NGF appeared to have an inhibitory effect on CL volume in pregnant cows, results are still pending for quantification of P, insulin-like growth factor 1, pregnancy-specific protein B, and interferon-stimulated genes, which will more accurately assess the effects that NGF may have on conceptus development. The results of this study will help us to better understand the role of the seminal plasma protein, NGF, at ovulation and determine if it can be utilised to enhance insemination programs in cattle.