Circulating P4 Concentrations Research Articles

Effect of 200 µg of gonadorelin at the first gonadotropin-releasing hormone of the Resynch-25 on ovarian dynamics and fertility in lactating Holstein cows

Our objective was to determine the effect of a 200-µg dose of GnRH 25 d after previous artificial insemination (AI) in a Resynch-25 resynchronization program on ovulatory response, circulating progesterone (P4) concentrations before and after treatment, and pregnancy per AI (P/AI) compared with a 100-µg dose in lactating Holstein cows. Experimental d 0 was considered the day of the previous AI. Lactating dairy cows (n = 3,240) with an average of 126 d in milk (DIM) and between 1 and 6 services were randomly assigned to receive 100 µg or 200 µg of GnRH on d 25 (GnRH25). On d 32 after AI, cows diagnosed nonpregnant with the presence of a corpus luteum (CL) detected by ultrasound (n = 1,249) received PGF2α treatments on d 32 and 33, followed by a GnRH 32 h later and AI 16 h after this last GnRH. Blood samples were collected on d 25, 32, and 34 to evaluate serum P4 concentrations. Transrectal ultrasonographic examination was performed on d 25 and 27 to assess ovulatory response to GnRH25. Cows were checked for pregnancy on d 32, 46, and 88 after AI. The larger dose of GnRH increased the overall proportion of cows that ovulated to the GnRH25 (25.0% for the 100-µg dose vs. 32.5% for the 200-µg dose). However, when cows were evaluated separately according to the pregnancy status on d 32 after AI, we found no treatment effect within cows pregnant and nonpregnant. Even though treatment increased the proportion of cows with serum P4 ≤0.42 ng/mL at the last GnRH treatment (G2; 86.2% for the 100-µg dose vs. 93.0% for the 200-µg dose), it did not affect P/AI on d 32, 46, and 88. Furthermore, a greater proportion of cows without a functional CL at GnRH25 had circulating P4 concentrations ≥1.00 ng/mL on d 32 and lower than 0.42 ng/mL on G2. These cows also had a greater P/AI on d 32, 46, and 88. In summary, the larger dose of GnRH on d 25 after AI did not increase the ovulatory response in nonpregnant cows and P/AI on d 32, 46, and 88 after AI after the Resynch-25 program. Additionally, nonpregnant cows without a functional CL at GnRH25 were better synchronized after the Resynch-25 protocol and had greater P/AI on d 32, 46, and 88 after timed-AI.

Influence of GnRH analog and dose on LH release and ovulatory response in Bos indicus heifers and cows on day seven of the estrous cycle

This study evaluated the influence of GnRH analogs (gonadorelin vs. buserelin) and doses (single vs. double) on LH release and ovulatory response in Bos indicus (Nelore) females on Day 7 of the estrous cycle. Cycling heifers and non-lactating cows were pre-synchronized: Day −10: progesterone (P4) implant insertion plus 2 mg of estradiol benzoate; Day −2: implant removal and 0.53 mg of cloprostenol sodium (PGF); Day 0: 25 μg of lecirelin (GnRH). Over four replicates, heifers (n = 57) and cows (n = 53) that ovulated to the GnRH treatment on Day 0, having a visible corpus luteum (CL) and a dominant follicle (DF) ≥ 8.5 mm, were allocated to receive the following GnRH treatments on Day 7: G-Single (100 μg of gonadorelin); G-Double (200 μg of gonadorelin); B-Single (10 μg of buserelin); and B-Double (20 μg of buserelin). At GnRH treatment, a P4 implant was inserted in heifers (0.5 g) and cows (1 g). Ultrasound examinations were done on Days −10, −2, 0, 2, 7, 9, 12, and 14 to evaluate DF diameter, ovulation and presence of CL. Blood samples were collected on Day 7 at 0, 2, and 4 h from GnRH treatment, to evaluate circulating P4 and LH concentrations. On Day 12, the P4 implant was removed, females received two PGF treatments (24 h apart), and 2 d later, 25 μg of GnRH was given to start the next replicate. In both heifers and cows, P4 concentrations were elevated on Day 7, and similar among groups (3.9 and 4.2 ng/mL, respectively). In heifers, buserelin induced greater LH peak (9.5 vs. 2.6 ng/mL; P < 0.01) and greater ovulation (88.9 [24/27] vs. 16.7% [5/30]; P < 0.01) than gonadorelin treatments, regardless of the dose. Similarly, in cows, buserelin induced greater LH peak than gonadorelin (9.9 vs. 4.9 ng/mL; P < 0.01). However, ovulation was only increased in cows from the B-Double group (90.9% [10/11]), whereas in the other groups the ovulatory response was similar (35.7% [15/42]). Regardless of treatment, heifers had similar P4 concentrations (P = 0.22), but smaller DF (P < 0.01) than cows on Day 7. Only in G-Double group the LH peak was lower (P = 0.05) in heifers than in cows, with no difference within other groups. In heifers, but not in cows, the single dose of buserelin resulted in high ovulatory response, equivalent to that produced by the double dose. In conclusion, in Bos indicus heifers and cows on Day 7 of the cycle, with elevated P4 concentrations, buserelin induced greater LH release and ovulatory response than gonadorelin treatments. Double doses increased the LH release, however, only resulted in greater ovulation in females treated with buserelin. Finally, although circulating P4 concentrations did not differ between parities, heifers were more likely to ovulate in response to a GnRH-induced LH peak than cows.

FRI419 Progesterone Regulation Of The GnRH Pulse Generator

Abstract Disclosure: E.G. Wall: None. A group of kisspeptin neurons located in the arcuate nucleus (ARN) of the hypothalamus represent the GnRH pulse generator and their activity is thought to be controlled primarily by the feedback actions of estradiol and progesterone (P4). In a healthy menstruating woman, following the mid-cycle ovulation, P4 levels dramatically increase and exert a negative feedback action onto the hypothalamus to slow pulsatile LH secretion. However, the exact mechanism by which P4 negatively feeds back to the hypothalamus remains unknown. The ARN kisspeptin neurons display synchronized episodes (SEs) of activity approximately every hour for most of the cycle but these dramatically reduce in number following the LH surge. As these ARN kisspeptin neurons express the progesterone receptor (PR) it is hypothesized that they may be the site at which P4 negative feedback occurs. Given the lack of detailed information in the mouse, we first assessed the profile of circulating P4 levels in mice at 8-hr intervals throughout the estrous cycle. Terminal blood samples were taken from individual mice along the 4 stages of the estrous cycle at 3-time points 8 hrs apart and the gonadal hormones were analyzed using LC-MS. The concentration of P4 was found to peak around the time of the LH surge at 6 pm on proestrus (21.94 ± 3.78 ng/ml) and was at its lowest at diestrus 10 am (0.42 ± 0.11 ng/ml). We next assessed the impact of circulating P4 on the activity of the ARN kisspeptin pulse generator using GCaMP fibre photometry in freely behaving mice. A prior study found that IP administration of 8 mg/kg P4 resulted in circulating P4 concentrations reaching 34 ng/ml (Wong et al., J Pharm Pharmacol 64, 2012). Hence, in this study, diestrus mice equipped for monitoring the activity of the kisspeptin pulse generator were injected with either 8 or 4 mg/kg P4 at 10 am and SEs monitored for 24 hrs. Both doses resulted in a significantly decreased frequency of SEs compared to the vehicle (p&amp;lt;0.05), which persisted for 6 hrs. To next examine whether P4 may be acting directly on the ARN kisspeptin neurons to slow pulse generation, P4 (25 ng/ml) was infused directly into the ARN while the activity of the pulse generator was monitored. Again, this resulted in a decreased frequency of SEs compared to the vehicle with the effects lasting approximately 5 hrs (p&amp;lt;0.05).These results indicate that P4 exerts powerful inhibitory control over the activity of the kisspeptin pulse generator and that these effects may be direct. Elucidating the mechanisms of P4 negative feedback may provide a platform for the development of new therapeutic strategies in the infertility clinic. Presentation: Friday, June 16, 2023

31 Correlation of the Reproductive Microbiome and Cytokine Profiles with Plasma Progesterone Concentrations in Postpartum Beef Cows Prior to Timed Artificial Insemination

Abstract Female sex steroid hormones have been shown to interact with cytokines within the immune system to help influence fertility. Previous research has indicated that shifts in commensal bacteria are associated with compromised reproductive performance during the postpartum period, yet evaluation of how the female sex hormone progesterone (P4) influences these microbial shifts is not fully understood. The objectives of this study were to determine 1) the relationship between P4 concentrations and cytokine concentrations, and 2) the relationship between P4 concentrations and the reproductive microbiome. Twenty Bos indicus-influenced beef cows (~60 days postpartum and free of health issues) were weighed, body condition scored and then subjected to the Bos indicus prostaglandin 5 day (d) + CIDR protocol beginning on d -8 and timed artificially inseminated on d 0. Blood samples were collected on d-3, -1, 0, and 28. Cytokines and reproductive microbiome were evaluated using vaginal and uterine flushes collected on d -3 and d-1. Experimental groups were assigned by d 28 pregnancy status, determined by transrectal ultrasonography, resulting in Open (n = 13) and Pregnant (n = 7) females. Bacterial community analyses were conducted targeting the V4 hypervariable region of the 16S rRNA gene. Plasma P4 concentrations on d -3 and d -1 were quantified per manufacturers’ instructions using a double-antibody RIA kit. Cytokine analyses of plasma and flushes were conducted using the RayBiotech Quantibody Bovine Cytokine Array Q1 kit per the manufacturer’s instructions. Statistical analyses were conducted using the GLM procedure and CORR procedure in SAS. Plasma P4 concentrations did not differ by pregnancy status (P &amp;gt; 0.05) but did decrease from d -3 to d -1 (0.89 vs. 0.08 ± 0.23 ng/mL, respectively; P = 0.02). There were no significant correlations between plasma P4 and plasma cytokines (P &amp;gt; 0.05). Interferon-alpha (IFNα) and P4 concentrations were positively correlated in Open and Pregnant uterine flushes (r = 0.41; P = 0.01). Concentrations of P4 interleukin (IL)-1F5 were negatively correlated in Open and Pregnant vaginal flushes (r = -0.32; P = 0.04). The relative abundance of genus Methanobrevibacter in uterine flushes was positively correlated with P4 concentrations in both experimental groups (r = 0.35; P = 0.02). Within vaginal flushes, there was a tendency for a negative correlation between the relative abundance of genus Bacteroides and P4 concentrations (r = -0.31; P = 0.05). Alternatively, there is a positive correlation between P4 concentration and relative abundance of genus Streptococcus (r = 0.47; P = 0.01) and genus Methanobrevibacter (r = 0.37; P = 0.02) in vaginal flushes. These results suggest there may be relationships between several cytokines and circulating P4 concentrations; therefore, potentially suggesting immunomodulatory activities of P4. Overall, this research alludes to P4 having a vital role in reproductive microbial shifts and associated immune responses. Further investigation on the local effects of P4 within the reproductive tract on immune responses and bacterial shifts could provide greater insight into mechanisms to improve fertility in beef cattle.

Luteal Function, Biometrics, and Echotextural Attributes in Santa Inês Ewes Superovulated with Different Total Doses of Porcine Follicle-Stimulating Hormone

Premature regression of corpora lutea (PRCL) may adversely affect the outcome of hormonal ovarian superstimulation in small ruminants, and the total dose of exogenous gonadotropins used may be one of the causes of this condition. There were two major objectives of the present study: (1) to evaluate the effects of different superovulatory doses of porcine follicle-stimulating hormone (pFSH) on the biometry, blood perfusion (Doppler), and echotextural characteristics of luteal structures; and, (2) to determine the usefulness of biometric, vascular, and echotextural luteal variables, as well as measurements of circulating progesterone (P4) concentrations for early detection of PRCL in superovulated Santa Inês ewes. Twenty-seven Santa Inês ewes received an intravaginal P4-releasing device (CIDR) from Days 0 to 8 (Day 0 = random day of the anovulatory period). An IM injection of d-cloprostenol (37.5 μg) was given at the time of the CIDR insertion and withdrawal. On Day 6, all the ewes received 300 IU of eCG IM and were divided into three treatment groups (each n = 9): G100 (100 mg); G133 (133 mg); and G200 (200 mg of pFSH) administered IM every 12 h in eight injections. Transrectal ovarian ultrasonography and jugular blood sampling for serum P4 measurements were performed on Days 11 to 15. On the day of embryo recovery (Day 15), all the ewes underwent diagnostic videolaparoscopy and were classified, based on their luteal characteristics, into three response groups: nCL (ewes with normal CL only); rCL (ewes with regressing CL only); and ewes with both nCL and rCL following the superovulatory regimen. Our present results indicate that the total pFSH doses of 100 mg and 200 mg result in similar ovulatory responses and luteal function/biometrics, although the percentage of donor ewes with nCL was greater (p < 0.05) for G100 compared with the G200 animals. An application of 133 mg of pFSH was associated with diminished luteogenesis. Lastly, circulating P4 concentrations, ultrasonographic estimates of total luteal area, and CL pixel heterogeneity (standard deviation of numerical pixel values) are promising markers of luteal inadequacy in superovulated ewes.

Luteal function in cyclic goats treated with human chorionic gonadotropin administered by intramuscular or intravaginal routes at the time of artificial insemination.

Human chorionic gonadotropin (hCG) has been used to improve goats reproductive efficiency. This study aimed to (i) evaluate if hCG administered by the intramuscular (i.m.) or intravaginal (i.vag.) route can be detected by a rapid β-hCG test in blood plasma samples and (ii) document ovarian effects of hCG administered by both routes at the time of artificial insemination (AI) performed 60 h after oestrus synchronization in goats. Twenty-two Alpine goats received two i.m. injections of 30 μg of d-cloprostenol (Prolise®, Tecnopec, São Paulo, Brazil) 7.5 days apart. One day after the onset of oestrus (at the time of AI), the goats were randomly allocated to one of the three groups that received: control (n=7): 0.3ml of saline solution intravaginally; hCGi.m. (n=7): 300 IU of hCG (Vetecor®; Hertape-Calier, São Paulo, Brazil) i.m. and hCGi.vag. (n=8): 300 IU of hCG deposited intravaginally. Blood samples were drawn at -1, 3, 6, 9 and 24 h after as well as on days 3, 7, 10, 13, 17 and 21 after hCG treatment/AI. All animals tested negative for hCG (ECO Diagnóstica, Corinto, Brazil) at -1h, and all control animals tested negative throughout the entire blood collection period. All hCGi.m. animals tested positive from 3h until D3 post-AI but only 50% of hCGi.vag. goats tested positive during the present study. In all animals studied, mean circulating P4 concentrations increased (p < .05) from D3 to D7 after AI and then declined (p < .05) from D10 to D17 in control and hCGi.m. groups and from D17 to D21 in the hCGi.vag. group. Total cross-sectional luteal area (CLA), mean colour Doppler area (DA), DA/CLA, mean high-velocity Doppler area and HVDA/CLA all declined (p < .05) by D17-D21 in all animals studied. In summary: (i) human chorionic gonadotropin could consistently be detected in blood samples using the rapid β-hCG test only in the hCGi.m. group; and (ii) there were no significant differences in the mean pregnancy rate, circulating P4 concentrations and various luteal parameters studied among Control, hCGi.m. and hCGi.vag. dose.

Administration of Coumestrol and/or Dimethyl Sulfoxide Affects Vaginal Epithelium and Sex Hormones in Female Dog

Background: The overpopulation of free dogs represents a risk to public health in some countries. However, current strategies for its control are insufficient; it is convenient to devise new alternatives. Methods: To determine the effects of a single oral administration of coumestrol (COU) on vaginal epithelial cells, serum progesterone and estradiol levels and clinical parameters: periovulatory bitches received a biscuit that contained 600 µg COU/kg diluted in DMSO or with DMSO alone or without additives. Each group was subdivided into animals that had ovulated on day 0 and those that had not. Result: In animals that ovulated, COU did not affect hormonal profiles. In contrast, animals that did not ovulate showed lower circulating P4 concentrations on days 21 and 28 after treatment than the control group. Furthermore, COU reduced parabasal cells and increased superficial anucleate cells. COU and DMSO are estrogenic and their actions depend on the ovulatory status of the animals.

Progesterone release profile and follicular development in Nelore cows receiving intravaginal progesterone devices

This study aimed to evaluate the progesterone (P4) release profile provided by four commercially available intravaginal P4 devices, as well as the effect of circulating P4 concentrations exclusively from these devices on the development of the dominant follicle (DF) in Nelore (Bos indicus) cows. Therefore, non-lactating multiparous Nelore cows were enrolled in an experimental design, over three replicates, starting on Day −9 with the insertion of a reused P4 device (2 g – original P4 load) for 7 d, followed by two treatments of cloprostenol sodium (PGF; 0.482 mg), 24 h apart, on Days −3 and −2. Just before device removal, on Day −2, a norgestomet ear implant was inserted and, 2 d later (Day 0), at the time of norgestomet withdrawal, cows were randomly assigned to receive one of the intravaginal devices: Primer (0.5 g); Prociclar (0.75 g); Sincrogest (1 g); or CIDR (1.9 g), and 2 mg of estradiol benzoate (EB) im. Blood samples were collected immediately before P4 device insertion, 12 h later and daily over 15 d (1 d after P4 device removal). Ultrasound examinations were performed on Days 0, 7, 8, 9, 10, 12, and 14 to evaluate ovarian dynamics. Results are presented as mean ± SEM and differences were considered when P ≤ 0.05. Overall, the devices resulted in distinct circulating P4 concentrations over 10 d, varying according to their initial P4 load and P4 impregnated surface area. Primer provided the lowest circulating P4 concentrations over time, whereas, CIDR had the greatest concentration. Sincrogest and Prociclar were similar, producing intermediary circulating P4. There was no effect of treatment on the DF diameter on any specific day, nor on follicular growth rate from Day 7–10. However, the Primer device resulted in a greater mean DF diameter over time. Additionally, greater circulating P4 concentrations, mainly during the first 3 d of device insertion, were associated with smaller DF diameters regardless of the treatment. In conclusion, results from this study provided a better understanding of the P4 profile of intravaginal P4 devices, as well as, their effect on DF development in Bos indicus cows. These data contribute to optimize the use of P4 devices in the reproductive management of beef cattle.

Progesterone release profile and follicular development in Holstein cows receiving intravaginal progesterone devices

The aim of this study was to evaluate the progesterone (P4) release profile provided by eight commercial intravaginal P4 devices, as well as the effect of circulating P4 concentrations produced exclusively by these devices on the development of the dominant follicle (DF) in non-lactating multiparous Holstein cows. All cows were submitted to the same experimental design starting with the insertion of a reused P4 device (2 g – original P4 load) for 7 d, followed by two treatments of cloprostenol sodium (PGF; 0.482 mg), 24 h apart, 6 and 7 d after device insertion. Just before device removal, a Norgestomet ear implant was inserted and, 2 d later (Day 0), simultaneously to Norgestomet withdrawal, cows received one of the tested intravaginal devices and 2 mg of estradiol benzoate (EB) im. In Exp.1 (n = 22; three replicates), cows were randomized to receive: CIDR (1.38 g); PRID-Delta (1.55 g); Prociclar (0.75 g); or Repro sync (2 g). In Exp. 2 (n = 29; four replicates), cows were randomized to receive: Cue-Mate (1.56 g); DIB 0.5 (0.5 g); DIB (1 g); PRID-Delta (1.55 g); or Sincrogest (1 g). Blood samples were collected before P4 device insertion (Day 0), 12 h later and daily over 15 d (1 d after P4 device removal). Ultrasound examinations were performed to evaluate growth of the DF on Days 0, 7, 8, 9, and 10. Results are presented as mean ± SEM and differences were considered when P ≤ 0.05. Overall, the circulating P4 profile and mean circulating P4 over 10 d differed among treatments. However, no effects were observed on the DF diameter and follicular growth rate from Day 7–10 after P4 device insertion. In Exp. 2, devices that provided higher circulating P4 concentrations were associated to a slower DF growth during the treatment period. Finally, this study provided a better understanding of the P4 release profile produced by intravaginal P4 devices as well as their effect on circulating P4 concentrations and DF development in non-lactating Holstein cows.

Effect of PGF2α treatments during early corpus luteum development on circulating progesterone concentrations and ovulation in breeding-age Holstein heifers

The objective of this study was to test the effect of low circulating concentrations of progesterone (P4) on pre-ovulatory follicle development in heifers as part of an overarching objective to develop a model to understand this phenomenon in dairy cattle without the confounding factors of lactation. Holstein heifers between 12 and 13 mo of age were pre-synchronized to ensure all heifers were on d 6 of the estrous cycle at the start of the Ovsynch program. Only heifers with CL regression and ovulation to the following pre-treatment strategy were used in the study: 0.5 mg cloprostenol (PGF2α), 2 d later, 0.1 mg GnRH, 6 d later, GnRH (G1; 1st GnRH of Ovsynch). Heifers (n = 159) responding to pre-treatment were randomly assigned to 4 groups and completed the Ovsynch program: high P4 control (HPC), low P4 control (LPC; PGF2α 24 h after G1), PG2 (PGF2α 24 and 48 h after G1) and PG3 (PGF2α 24, 48, and 96 h after G1). Only heifers that had ovulation to G1 remained in the study. Blood samples were collected in all heifers on d 7 (n = 157) and in a subset of heifers on d 1, 2, 3, 4 (n = 82) after G1 to measure serum P4. Pre-ovulatory follicle size at G1 (13.0 ± 0.1 mm; P = 0.53) and mean serum P4 24 h after G1 (d 1; 3.62 ± 0.11 ng/mL; P = 0.46) did not differ among treatments. HPC heifers had greater (P < 0.001) mean serum P4 compared to LPC, PG2 and PG3 on d 2, 3, 4, and 7. On d 2, 3 and 4, mean serum P4 of LPC, PG2 and PG3 heifers did not differ (P > 0.10). On d 7, LPC heifers had greater (P < 0.001) serum P4 compared to PG2 and PG3 heifers. Mean ± SEM serum P4 on d 7 after G1 was 8.43 ± 0.39, 2.55 ± 0.36, 1.58 ± 0.20, and 1.21 ± 0.15 ng/mL for HPC, LPC, PG2, and PG3, respectively. Percentage of heifers with P4 < 0.50 ng/mL on d 7 was greater (P < 0.05) for LPC, PG2 and PG3 (27, 32 and 26%, respectively) compared to HPC (0%). A greater (P < 0.05) proportion of heifers ovulated before G2 in the LPC, PG2 and PG3 than in the HPC. For heifers that ovulated after G2, low serum concentrations of P4 in LPC, PG2 and PG3 induced double ovulations in 6/97 heifers after the final GnRH of Ovsynch compared to 0/33 in HPC. In summary, PGF2α treatments during early CL development reduced circulating P4 concentrations 7 d after G1 compared with both HPC and LPC. However, it did not effectively control CL and follicle function to be utilized as a model to test high vs. low serum P4 on fertility parameters in Holstein heifers.

Effect of types of breeding on embryo survival following first AI in lactating Holstein cows.

The main objective was to investigate the effects of timed-AI protocols versus AI following oestrus detection on circulating progesterone (P4) and embryo survival after first service in Holstein cows. Cycling status was determined by ultrasonography and by plasma P4 concentrations 14 and 26days after calving, and only cows with a corpus luteum and/or P4≥1ng/ml were used. Cows were randomly allocated to one of three types of breeding: DO (n=80), received GnRH-7d-PGF2α-3d-GnRH and Ovsynch56 was initiated 7days later; G7G (n=70), received PGF2α-2d-GnRH and Ovsynch56 (GnRH-7d-PGF2α-56h-GnRH-16h-AI) was initiated 7days later; or AI based on oestrus detection, EDAI (n=60). Progesterone was also determined at AI and 8, 16, 18 and 20days after AI; ISG15 and MX2 mRNA abundance were determined 16days after AI. Mean plasma P4 at AI was greater in the EDAI group compared with DO and G7G groups, while after AI, P4 was greater in DO and G7G groups compared with EDAI group. However, the percentage of cows with a concentration of P4<0.8ng/ml at AI did not differ among groups. Relative mRNA abundance of ISG15 and MX2 was greater in the DO and G7G groups compared to those in EDAI group. Pregnancy per AI 16, 32 and 60days after AI was greater (p<.05) in cows in the DO group compared with those in EDAI group (47.5%, 38.8% and 36.3% vs. 30.0%, 21.7% and 15.0%). Pregnancy losses between 16 and 60 days after AI were greater (p<.05) in cows in the EDAI (50.0%) group compared to those subjected to DO (23.7%) or G7G (24.1%). In conclusion, the use of timed-AI synchronization protocols resulted in greater circulating P4 concentrations post-AI and greater embryo survival following first service in lactating Holstein cows.

Circulating progesterone concentrations and preovulatory follicle diameters affecting ovulatory response in crossbred dairy heifers, following a 7-day progesterone-based synchronization protocol.

The present study was aimed at evaluating the differences of ovarian follicular dynamics and circulating progesterone (P4) concentrations between crossbred Holstein heifers that ovulated and did not ovulate after a P4-based synchronization protocol. Twenty-one crossbred (Holstein × Thai native) heifers with random stages of the oestrous cycle were subjected to the ovulation synchronization protocol, using an intravaginal P4-releasing device (Eazi-Breed CIDR®) for 7 days. Out of 21 CIDR-treated heifers, 14 ovulating heifers were classified as the ovulatory group and 7 non-ovulating heifers were considered the anovulatory group. The heifers having new wave emergence in ovulatory and anovulatory groups were 11/14 (78.6%) and 4/7 (57.1%), respectively. In ovulating heifers, the mean (± SEM) diameter of preovulatory follicle (PF, mm) was significantly larger, compared to non-ovulating heifers (7.21 ± 0.32 versus 4.04 ± 0.44; P = 0.001), while the mean (± SEM) follicular growth rates (mm/d) in non-ovulating heifers tended to be lower, compared to ovulating heifers (0.73 ± 0.17 versus 1.06 ± 0.08; P = 0.07). The mean (± SEM) circulating P4 concentration (ng/ml) throughout the CIDR protocol (0-10 days) in non-ovulating heifers was significantly higher, in comparison with ovulating heifers (2.82 ± 0.27 versus 1.83 ± 0.16; P = 0.02). However, no significant difference in the mean corpus luteum volume between groups was observed. In conclusion, the present results suggested that elevated circulating P4 concentrations and smaller PF diameters could cause ovulation failure in crossbred Holstein heifers, following a 7-day CIDR-based synchronization protocol.

PSII-28 Effect of route of administration of dinoprost tromethamine on serum profiles of PGFM and progesterone in lactating Holstein cows

Abstract Our objective was to compare serum profiles of 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM) and circulating progesterone (P4) concentrations in lactating Holstein cows based on route of administration of dinoprost tromethamine administered 7 d after administration of GnRH. Multiparous lactating Holstein cows were fitted with indwelling jugular catheters 6 d after the last GnRH treatment of an Ovsynch protocol (d 0), and cows were randomized on d 7 to receive 25 mg dinoprost tromethamine (2 mL Lutalyse® HighCon; Zoetis, Parsippany-Troy Hills, NJ) either s.c. in the neck (SC; n = 6) or i.m. in the semitendenous muscle (IM; n = 6). Blood samples were collected via jugular catheters 24, 2, and 1 h before treatment to establish baseline P4 concentrations. Blood samples were collected every 15 min after treatment for 1.75 h, then every 30 min for 48 h, and at 60 and 72 h with the last time point corresponding to timed AI after an Ovsynch protocol. Serum samples were stored frozen and later assayed for PGFM and P4 concentrations via ELISA. Circulating PGFM concentrations were greater (P &amp;lt; 0.05) for SC than for IM cows from 15 to 90 minutes after treatment which resulted in a greater (P = 0.02) area under the PGFM curve during the first 2 h after treatment (2,223 ± 160 pg•hr/mL vs. 1,586 ± 150 pg•hr/mL for SC vs. IM cows, respectively). By contrast, circulating P4 concentrations normalized to percent of baseline P4 concentrations did not differ between treatments at any time point after treatment during the experiment. In conclusion, although SC cows had greater circulating PGFM concentrations 15 to 90 min after treatment than IM cows, the decrease in circulating P4 concentrations during induced luteolysis did not differ based on route of dinoprost tromethamine administration. Supported by NIFA USDA Hatch project 1019532.

Measurements of circulating progesterone and estrone sulfate concentrations as a diagnostic and prognostic tool in porcine pregnancy revisited

The main goal of this study was to examine the utility of measuring systemic concentrations of steroid hormones, namely progesterone (P4) and estrone sulfate (E1S), for monitoring the progression of porcine pregnancy and predicting sow fertility. There were 3 subsets of artificially inseminated (AI'd) sows used in the present experiments: (i) animals sacrificed on gestational day 20 (gd20; n = 16) or (ii) gd50 (n = 16; Experiment 1), and (iii) animals maintained throughout pregnancy (n = 24; Experiment 2). Blood samples (10 mL) were drawn from the orbital sinus and the endocrine data determined at different time points around ovulation/artificial insemination (gd0 (first AI), gd1 (second AI), and gd2) and maternal recognition of pregnancy (gd11), as well as on gd20 and gd50 (during 2 periods of increased embryonic/fetal mortality in swine) were examined for correlations with the numbers of healthy, arrested, and reabsorbing embryos (Experiment 1) or with the number of live, stillborn, and mummified piglets recorded at farrowing (Experiment 2). No correlations were recorded between circulating concentrations of both steroids and the numbers of healthy, arresting, or reabsorbing conceptuses on gd20 or 50 (Experiment 1). The number of corpora lutea (CL) was directly related to the number of healthy embryos/conceptuses on gd20 and 50 (r = 0.71, P = 0.007 and r = 0.76, P = 0.0007, respectively) and the number of arresting embryos on gd20 (r = 0.54, P = 0.05), and negatively correlated with the number of reabsorbing embryos on gd20 (r = −0.53, P = 0.05). In Experiment 2, circulating P4 concentrations on gd11 related directly to the number of live-born piglets (r = 0.46, P < 0.04). Systemic E1S concentrations on gd0, gd1, gd2 and gd50 were correlated with the number of mummified conceptuses recorded at farrowing (r = 0.50, P = 0.03; r = 0.59, P = 0.01; r = 0.48, P = 0.04; and r = 0.56, P = 0.01, respectively) and plasma concentrations of E1S on gd20 related directly to the number of stillborn piglets (r = 0.60, P = 0.02). In summary, the number of CL on gd20 and 50 is a reliable marker of embryonic/fetal pig status. Measurements of P4 and E1S on gd20 and 50 showed limited diagnostic value (ie, were not indicative of the number of healthy and abnormally developing embryos/fetuses). However, measurements of circulating P4 and E1S concentrations during the periconceptional period and in the early/mid-pregnancy of sows have the makings of a practical method to predict gestational outcomes.

Hepatic mRNA expression of enzymes associated with progesterone metabolism and its impact on ovarian and endocrine responses in Nelore (Bos indicus) and Holstein (Bos taurus) heifers with differing feed intakes

The aim of this study was to evaluate circulating progesterone concentration (P4), LH pulsatility and ovarian follicular dynamics in Nelore (B. indicus) and Holstein (B. taurus) heifers under high (HDMI) and low (LDMI) dry matter/energy intakes. In addition, the effects of dry matter/energy intake and breed on hepatic expression of six genes associated with P4 metabolism (AKR1C4, AKR1D1, CYP3A4, CYP2C19, SRD5A1, and SRD5A3) was evaluated. Heifers received an intravaginal P4 device (1 g), 2 mg of estradiol benzoate (EB) i.m. and 500 μg of PGF2α at the begging of the synchronization protocol (D0). Eight days later, the P4 device was removed and all heifers received 1 mg of EB 24h later. Regardless of dry matter/energy intake, the number of recruited follicles was greater in Nelore than in Holstein heifers. In contrast, the maximum diameter of the dominant follicle was greater in Holstein than in Nelore heifers. Circulating P4 concentrations were greater in Nelore than in Holstein from D2 to D9, and in heifers receiving LDMI than those receiving HDMI from D1 to D8 of hormonal protocol. In addition, Holstein heifers had greater LH pulsatility and area under the curve of LH peaks compared to Nelore heifers. However, no effects were observed for LH values between feed intake levels. Interestingly, Holstein heifers had higher expression of SRD5A1, AKR1C4, AKR1D1 than Nelore heifers; whereas, for Nelore heifers, only the expression of CYP3A4 was higher compared to Holstein heifers. In conclusion, there are important differences in the follicular dynamics, circulating P4 and LH pulsatility concentrations that need to be considered during synchronization protocols for Nelore and Holstein breeds. More importantly, these differences appear to be at least partially modulated by the level of feed intake and the contrasting enzyme system in the liver involved with P4 metabolism between these cattle breeds.

Evaluation of presynchronization and addition of GnRH at the beginning of an estradiol/progesterone protocol on circulating progesterone and fertility of lactating dairy cows

The objectives of this study were to determine if the utilization of a presynchronization strategy would improve fertility at first artificial insemination (AI) during an E2/P4 ovulation synchronization protocol with or without GnRH administration at the beginning of the protocol. This experiment was conducted using cows (n = 665) at their first postpartum service and the following breeding treatment: CIDR insertion and 2.0 mg of estradiol benzoate (EB) on day −11; 25 mg dinoprost tromethamine (PG) on day −4; PG, CIDR withdrawal, and 1.0 mg of estradiol cypionate (ECP) on day −2; timed-AI on day 0. At 31 ± 3 days postpartum, cows were randomly allocated to one of three treatments on a weekly basis: 1) -P + GnRH: cows assigned to the breeding protocol with 100 μg of GnRH on day −11, 2) P + GnRH: cows assigned to a presynchronization protocol using CIDR insertion +2 mg EB on day −28, PG + ECP and CIDR withdrawal on day −21, and beginning of the breeding protocol plus GnRH (100 μg) on day −11, and 3) +P-GnRH: cows assigned to a presynchronization protocol and the breeding treatment without GnRH on day −11. No treatment effects were observed on P/AI at the pregnancy diagnoses on days 32 and 60, or for pregnancy losses between days 32 and 60 of pregnancy whether analyses included all cows or only cows that ovulated near TAI. Moreover, milk yield negatively affected P/AI. Cows with greater circulating P4 concentrations on day −4 had greater P/AI on day 60. Cows without CL on day −11 had a reduced P/AI and this effect was more significant in cows not treated with GnRH. Cows assigned to -P + GnRH had the lowest circulating P4 concentration on day −4 (3.4 ± 0.16 ng/mL), followed by + P-GnRH (4.56 ± 0.17 ng/mL), and +P + GnRH (5.08 ± 0.17 ng/mL) cohorts. The data of the current study suggest that the combination of a Presynch and GnRH administration at the beginning of a TAI protocol was the most effective way to increase the % of cows with a functional CL and with elevated circulating P4 concentrations at the time of PG treatment.

Effect of chronic administration of a gonadotropin-releasing agonist on luteal function and pregnancy rates in dairy cattle.

Increased embryonic losses may be associated with inadequate progesterone (P4) concentrations in high-producing lactating dairy cattle. The objectives of the present studies were to determine if chronic administration of a gonadotropin-releasing hormone (GnRH) agonist, Deslorelin, would increase circulating P4 concentrations and subsequently increase pregnancy rates in dairy cattle. Administration of Deslorelin for 12days increased (p<.05) luteal volume and circulating P4 concentrations in primiparous lactating dairy cows, but increased only luteal volumes in multiparous cows. Treatment with Deslorelin increased Day 45 pregnancy rates in cows as compared to untreated controls. Chronic treatment with Deslorelin in dairy cattle; (a) increased luteal volume of the primary CL, (b) induced accessory CL, (c) increased circulating P4 concentration in primiparous cows only, (d) did not lengthen the estrous cycle upon removal of treatment, and (e) increased pregnancy rates. Although luteal volume was increased in multiparous cows and circulating P4 concentrations were not with Deslorelin treatment, there was an apparent effect on pregnancy rates. This hormonal strategy may represent a suitable model to address local effects of P4 and GnRH/luteinizing hormone on uterine environment and subsequent embryonic survival.

Ultrasonography-accessed luteal size endpoint that most closely associates with circulating progesterone during the estrous cycle and early pregnancy in beef cows

The aim was to evaluate the associations between circulating P4 concentrations, corpus luteum (CL) size (diameter, area or volume) and blood perfusion (BP) in cows. In Experiment 1, Pearson’s correlations (P < 0.05) with P4 concentrations were observed during CL development (D8) for total area (TA; r = 0.76), luteal area (ACL; r = 0.72), total and luteal diameter (TD and DCL respectively; r = 0.46). During mid-late diestrus, there was a positive correlation (P < 0.05) only at D15 with TA and ACL (r > 0.60), TD, total volume (TV) and luteal volume (VCL; r > 0.434). During luteal regression, the correlation was only observed at D18 for ACL (r = 0.478) and D20 with several variables. In Experiment 2, CL weight and ACL had the greatest correlation with P4 (r > 0.6). In Experiment 3, TA and ACL were the variables that were most closely correlated with serum P4 concentrations at D7 in recipient cows. Correlation coefficients were greater for luteal measurements when there were compact compared with cavitary CLs. In Experiment 4, there was no correlation (P > 0.05) between P4 and any of the variables measured on D4 and D7 in recipient cows detected in estrus. On D18 to D20, all CL characteristics were correlated (P < 0.05) with plasma P4, and luteal BP and BP area were more closely (P < 0.05) correlated than ACL. In conclusion, CL perimeter area measurements had the greatest association with luteal function during CL development; whereas for BP there was a greater correlation with P4 than luteal size during luteolysis.

Profile of LH release in response to intramuscular treatment with kisspeptin in Bos indicus and Bos taurus prepubertal heifers

The main objective of this study was to evaluate the effect of intramuscular (I.M.) administration of different doses of kisspeptin (Kp) on the pattern of luteinizing hormone (LH) release in Bos taurus and Bos indicus prepubertal heifers. Holstein heifers weighing 215.1 ± 38.6 kg (n = 24; aged 5–8 mo) and Gyr heifers weighing 215.4 ± 31.4 kg (n = 24; aged 6–10 mo) were enrolled in this study. The animals were confirmed as non-cyclical by absence of a corpus luteum (CL) as determined by ultrasound scanning of the ovaries and low circulating P4 concentrations (<1.0 ng/mL) evaluated at a 10-day interval (on day −10 and day 0). For each genetic group, heifers were randomly assigned to one of four treatments: Kp at 2.5 μg/kg body weight (Kp2.5), Kp at 5.0 μg/kg (Kp5), Kp at 10 μg/kg (Kp10), or gonadotropin-releasing hormone (GnRH) agonist (0.01 mg of buserelin acetate per heifer), all administered by I.M. injection. All animals responded to the treatments with an LH surge (P < 0.01). There was an effect of breed (P < 0.01) on induced LH release, with Holstein heifers having a greater area under the curve for LH (AUC; P < 0.01) and greater LH peak amplitude (P < 0.01) than Gyr heifers. Nevertheless, greater AUC for LH occurred in Kp10 heifers for both breeds. There was no effect of breed on LH AUC or LH peak amplitude after GnRH agonist treatment. For both breeds, heifers treated with Kp displayed an earlier (P < 0.01) LH peak with a reduced amplitude of the LH peak (P < 0.01) and reduced LH AUC (P < 0.01) compared to heifers treated with GnRH agonist. Thus, both zebu and taurus immature heifers treated with I.M. Kp injection responded with a rapid and dose-dependent LH surge, although even large doses of the native Kp-10 did not mimic the magnitude or duration of the LH surge produced by the GnRH agonist. The early onset of the LH surge after Kp treatment and the short duration suggest that the effects of Kp were likely due to pituitary rather than hypothalamic action. Finally, there seems to be a greater responsiveness to Kp in Bos taurus than in Bos indicus prepubertal heifers, although LH release after GnRH agonist treatment was similar for the two breeds.

Randomised controlled trial of the effect of concentration of progesterone before artificial insemination on fertility in ovulatory and anovulatory Bos indicus cattle.

To investigate the effect of concentration of progesterone (P4) before artificial insemination (AI) on fertility in ovulatory or anovulatory Bos indicus cattle. Randomised control study METHODS: The study included 162 heifers and 96 lactating cows. On days -10 to -12, animals were examined using transrectal ultrasound, administered PG and examined for a corpus luteum (CL). Those with a CL were allocated to Experiment 1. On day 0 they were administered an intravaginal progesterone-releasing device (IVD) containing progesterone (P4) (0.78 g), oestrodiol benzoate (ODB) and either saline or PG to induce high and low circulating P4 concentrations, respectively. Those without a CL were re-examined on day 0 and those without a CL at both examinations were allocated to Experiment 2. Cows and heifers were treated with an IVD containing P4 at 0.78 g or 1.56 g to induce low or high P4 concentrations, respectively. IVDs were removed on day 7 and PG and equine chorionic gonadotrophin (eCG) were administered. Females in oestrus on day 9 were inseminated; others were administered ODB and inseminated 22-26 h later. Greater concentrations of circulating P4 increased the odds of pregnancy to AI in anovulatory females (P = 0.008), but decreased the odds of pregnancy in one year but not another in ovulatory animals (P × year, P = 0.019). Manipulating P4 concentrations before AI has the potential to improve pregnancy outcomes to AI in B. indicus females, but treatment may need to vary between animals classified as anovulatory or ovulatory.