Measuring Plasma Progesterone Concentrations Research Articles

Effect of various hCG treatment protocols on luteal characteristics, plasma progesterone concentration, and pregnancy in normal cyclic Indian crossbred dairy cows.

Limited information exists on the application of human chorionic gonadotropin (hCG) post insemination in the Indian crossbred dairy cows. This study aimed to evaluate the effect of four hCG administration protocols on luteal characteristics and pregnancy outcomes following artificial insemination (AI). Using block randomization, 50 healthy lactating crossbred cows were allocated in equal numbers (n = 10 cows per group) to receive either 5 mL normal saline (control) or 1500 IU hCG on the day of AI (hGG-0), day 7 post AI (hCG-7), day 14 post AI (hCG-14), or days 0, 7, and 14 post AI (hCG-0,7,14). All cows were scanned using sequential transrectal ultrasound examinations to evaluate primary luteal parameters, development of accessory corpora lutea, and pregnancy. Serial blood samples were collected to measure plasma progesterone concentrations. Data were analyzed using repeated measures ANOVA and Fisher's exact tests. The mean primary luteal area, total luteal area, and total luteal diameter values were significantly greater in the hCG-treated cows. Compared to the control, the hCG-14 group had a significantly higher percentage of cows with an accessory corpora luteum. However, there were no significant differences in the mean progesterone concentrations or the first service conception rates between any of the groups. Overall, the results of this study indicate that while hCG administration post AI in healthy Indian crossbred cows may enhance primary luteal dimensions or induce the formation of accessory corpora lutea, it does not appear to have any beneficial effect on luteal function or pregnancy.

Heifers with positive genetic merit for fertility traits reach puberty earlier and have a greater pregnancy rate than heifers with negative genetic merit for fertility traits

This study investigated the hypothesis that dairy heifers divergent in genetic merit for fertility traits differ in the age of puberty and reproductive performance. New Zealand's fertility breeding value (FertBV) is the proportion of a sire's daughters expected to calve in the first 42 d of the seasonal calving period. We used the New Zealand national dairy database to identify and select Holstein-Friesian dams with either positive (POS, +5 FertBV, n = 1,334) or negative FertBV (NEG, -5% FertBV, n = 1,662) for insemination with semen from POS or NEG FertBV sires, respectively. The resulting POS and NEG heifers were predicted to have a difference in average FertBV of 10 percentage points. We enrolled 640 heifer calves (POS, n = 324; NEG, n = 316) at 9 d ± 5.4 d (± standard deviation; SD) for the POS calves and 8 d ± 4.4 d old for the NEG calves. Of these, 275 POS and 248 NEG heifers were DNA parent verified and retained for further study. The average FertBV was +5.0% (SD = 0.74) and -5.1% (SD = 1.36) for POS and NEG groups, respectively. Heifers were reared at 2 successive facilities as follows: (1) calf rearing (enrollment to ∼13 wk of age) and (2) grazier, after 13 wk until 22 mo of age. All heifers wore a collar with an activity sensor to monitor estrus events starting at 8 mo of age, and we collected weekly blood samples when individual heifers reached 190 kg of body weight (BW) to measure plasma progesterone concentrations. Puberty was characterized by plasma progesterone concentrations >1 ng/mL in at least 2 of 3 successive weeks. Date of puberty was defined when the first of these samples was >1 ng/mL. Heifers were seasonally bred for 98 d starting at ∼14 mo of age. Transrectal ultrasound was used to confirm pregnancy and combined with activity data to estimate breeding and pregnancy dates. We measured BW every 2 wk, and body condition and stature at 6, 9, 12, and 15 mo of age. The significant FertBV by day interaction for BW was such that the NEG heifers had increasingly greater BW with age. This difference was mirrored with the significant FertBV by month interaction for average daily gain, with the NEG heifers having a greater average daily gain between 9 and 18 mo of age. There was no difference in heifer stature between the POS and NEG heifers. The POS heifers were younger and lighter at puberty, and were at a lesser mature BW, compared with the NEG heifers. As a result, 94 ± 1.6% of the POS and 82 ± 3.2% of the NEG heifers had reached puberty at the start of breeding. The POS heifers were 20% and 11% more likely to be pregnant after 21 d and 42 d of breeding than NEG heifers (relative risk = 1.20, 95% confidence interval of 1.03-1.34; relative risk = 1.11, 95% confidence interval of 1.01-1.16). Results from this experiment support an association between extremes in genetic merit for fertility base on cow traits and heifer reproduction. Our results indicate that heifer puberty and pregnancy rates are affected by genetic merit for fertility traits, and these may be useful phenotypes for genetic selection.

Prostaglandin F<sub>2α</sub> induced luteolysis is delayed but not prevented by acute administration of luteotropic drugs in lactating nonpregnant Holstein cows

Inadvertent use of prostaglandin F2α (PGF2α) in pregnant cattle could result in luteolysis and pregnancy loss. Using a nonpregnant cow model, we determined if luteotropic agents could counteract luteolytic effects of PGF2α. Ovarian status of 20 lactating nonpregnant Holstein cows was synchronized using an Ovsynch protocol and ovulation confirmed by transrectal ultrasonography 48 hours after the second gonadotropin releasing hormone (GnRH). Eight days after ovulation, 25 mg of dinoprost (native PGF2α) was administered IM to induce luteolysis. Five minutes after PGF2α, cows were treated IM with gonadorelin (GnRH, 100 μg; n = 5), human chorionic gonadotropin (hCG, 1,000 IU; n = 6), porcine pituitary luteinizing hormone (pLH, 25 mg; n = 5), or saline (control, 2 ml; n = 4). Blood samples were collected before PGF2α (0 hour) and at 1, 6, 12, 18, 24, 30, 36, 42, 48, 60, 72, and 84 hours after PGF2α to monitor subsequent luteal activity by measuring plasma progesterone concentrations. Although none of the treatments counteracted the luteolytic action of PGF2α, the rate of decline in plasma progesterone within 1 hour after PGF2α was greater (p = 0.04) in control than in GnRH, hCG, and pLH (2.2 versus 0.2, 0.3, and 0.1 ng/ml/hour, respectively). In addition, pLH-treated cows tended (p = 0.07) to have greater overall mean plasma progesterone for up to 84 hours (1.6 ± 0.2 ng/ml) than other treatments combined (1.1 ± 0.1 ng/ml). In the present study, giving GnRH, hCG or pLH 5 minutes after PGF2α administration was not effective in countering luteolytic effects of PGF2α. Nevertheless, pLH treatment tended to delay PGF2α induced luteolysis, which warrants further investigation.

The reproductive response to the male effect of 7- or 10-month-old female goats is improved when photostimulated males are used

The exposure of adult, female, Mediterranean goats during anoestrus to males with induced sexual activity via photostimulation, induces a very high percentage of ovulations. The present work examines the ability of photostimulated bucks to improve the male effect-induced reproductive response of young does over that induced by non-stimulated bucks. A 2×2 factorial experiment was designed, consisting of doe age and buck photoperiod treatments. During seasonal anoestrus, 41 does aged 7 (n=19) or 10 (n=22) months were subjected to the male effect on 10 April; half of each group was exposed to males rendered sexually active by prior exposure to 3 months of long days (16 h of light/day) from 31 October (PHOTO bucks), and half to males maintained under the natural photoperiod (CONTROL bucks). Oestrous activity was recorded daily by direct visual observation of the marks left by male-worn marking harnesses over the 32 days following the bringing of the sexes together (introduction). Doe body weight and body condition were determined weekly. Ovulation was detected by measuring plasma progesterone concentrations twice per week over the 3 weeks after introduction. The ovulation rate was assessed by transrectal ultrasonography. Fecundity, fertility, prolificacy and productivity were also determined. The interaction doe age × buck photoperiod treatment had no effect on any outcome. The percentage of females showing ovulation or oestrus was higher in the does exposed to PHOTO bucks (85% v. 43% for those exposed to CONTROL bucks) they also showed higher fertility (75% v. 43%) and productivity (1.05±0.17 v. 0.57±0.16 kids born per doe serviced) (all P values at least P<0.05). The 10-month-old group showed higher percentage of females showing ovulation, oestrus, fertility and productivity than the 7-month-old does after the male effect (females showing ovulation: 82% v. 42%; showing oestrus: 73% v. 42%; fertility: 73% v. 42% and productivity: 1.09±0.17 v. 0.47±0.14 goat kids born per doe serviced; respectively, all P values at least P<0.05). The present results show that the use of photostimulated males improves the reproductive performance of 7- and 10-month-old does, and may contribute towards increasing their productivity and lifetime reproductive performance.

Effects of varying doses of β-nerve growth factor on the timing of ovulation, plasma progesterone concentration and corpus luteum size in female alpacas (Vicugna pacos).

Ovulation in camelids is induced by the seminal plasma protein ovulation-inducing factor (OIF), recently identified as β-nerve growth factor (β-NGF). The present study measured the total protein concentration in alpaca seminal plasma using a bicinchoninic acid (BCA) protein quantification assay and found it to be 22.2±2.0mgmL(-1). To measure the effects of varying doses of β-NGF on the incidence and timing of ovulation, corpus luteum (CL) size and plasma progesterone concentration, 24 female alpacas were synchronised and treated with either: (1) 1mL 0.9% saline (n=5); (2) 4µg buserelin (n=5); (3) 1mg β-NGF protein (n=5); (4) 0.1mg β-NGF (n=5); or (5) 0.01mg β-NGF (n=4). Females were examined by transrectal ultrasonography at 1-2-h intervals between 20 and 45h after treatment or until ovulation occurred, as well as on Day 8 to observe the size of the CL, at which time blood was collected to measure plasma progesterone concentrations. Ovulation was detected in 0/5, 5/5, 5/5, 3/5 and 0/4 female alpacas treated with saline, buserelin, 1, 0.1 and 0.01mg β-NGF, respectively. Mean ovulation interval (P=0.76), CL diameter (P=0.96) and plasma progesterone concentration (P=0.96) did not differ between treatments. Mean ovulation interval overall was 26.2±1.0h. In conclusion, buserelin and 1mg β-NGF are equally effective at inducing ovulation in female alpacas, but at doses ≤0.1mg, β-NGF is not a reliable method for the induction of ovulation.

Dose-Response of Female Llamas to Ovulation-Inducing Factor from Seminal Plasma

The present study was designed to determine if the dose of purified ovulation-inducing factor (OIF) from llama seminal plasma required to provoke an ovulatory response is physiologically relevant in terms of the proportion present in a normal ejaculate and to test the hypothesis that corpus luteum (CL) form and function are affected by OIF in a dose-dependent manner. Female llamas were assigned randomly to five groups (n = 10 per group) and given a single i.m. dose of 500, 250, 125, or 60 μg of purified OIF (representative of the amount present in 1/25th to 1/200th of a normal ejaculate) or 1 ml of PBS (control). Ovulation and CL development were monitored by transrectal ultrasonography. Blood samples were taken to measure plasma progesterone concentrations and to determine changes in plasma concentrations of luteinizing hormone (LH). The high dose of OIF (500 μg) was associated with the highest incidence of ovulation (P < 0.05), the greatest maximum CL diameter (P < 0.05), and the largest day-to-day profiles of CL diameter (P < 0.05) and plasma progesterone concentrations (P < 0.01). A rise in plasma LH concentration was apparent in all llamas that ovulated and was most rapid and highest in the high-dose group (P < 0.01). The low dose of OIF (60 μg) was minimally effective for induction of ovulation and the least luteotrophic, as evidenced by the smallest maximum CL diameter and the smallest day-to-day profiles for CL diameter and plasma concentrations of progesterone and LH. Responses were intermediate for the middle-dose groups (125 and 250 μg). We conclude that OIF from llama seminal plasma has a dose-dependent effect on ovulation rate and CL form and function in llamas and that the biological effect of OIF is evident at physiologically relevant doses (i.e., as little as 1/100th of that present in an ejaculate).

Embryo transfer technique: Factors affecting the viability of the corpus luteum in llamas

The aim of the present study was to evaluate the effect of the embryo transfer (ET) maneuvers on plasma progesterone concentrations in recipient Lama glama females and the relationship between the site the embryo was transferred to and corpus luteum (CL) localization. Experiment I (effect of transcervical threading): adult non-pregnant, non-lactating llama females were randomly assigned into two groups: control group (without cervical threading, n = 10) and group A (with cervical threading, n = 10). In both groups, CL activity was evaluated through measurement of progesterone plasma concentrations. In group A, on Day 6 after inducing ovulation with buserelin, the cervix was threaded to evaluate the effect of the maneuver on CL viability. No significant differences were observed in mean progesterone concentrations between groups ( P > 0.05). Experiment II (effect of depositing PBS): females ( n = 66) were randomly assigned into six groups ( n = 10 per group and control group: n = 6) to evaluate the effect of depositing PBS in different sites in the uterus in relation to the localization of the CL: group ‘Left-Ipsilateral’: transcervical placing of PBS in the left uterine horn (CL in left ovary); group ‘Left-Contralateral’: transcervical placing of PBS in the left uterine horn (CL in right ovary); group ‘Right-Ipsilateral’: transcervical placing of PBS in the right uterine horn (CL in right ovary); group ‘Body-Left’: transcervical placing of PBS in the uterine body (CL in left ovary); group ‘Body-Right’: transcervical placing of PBS in the uterine body (CL in right ovary) and control group. Corpus luteum activity was evaluated in all groups by measuring plasma progesterone concentrations. On Day 6 post-buserelin, the corresponding maneuver was carried out according to the group. No significant differences were found for the mean plasma progesterone concentrations between groups ( P > 0.05). Experiment III (effect of ET on CL viability): females ( n = 22) were used as embryo donors and 50 females as recipients, in order to evaluate if placing the embryo in different areas of the uterus influences CL viability. Recipients were randomly divided into five groups, according to the place in the uterus where the ET was conducted with respect to the ovary where ovulation occurred: group ‘Left-Ipsilateral’: ET in the left uterine horn (CL in left ovary); group ‘Left-Contralateral’: ET in the left uterine horn (CL in right ovary); group ‘Right-Ipsilateral’: ET in the right uterine horn (CL in right ovary); group ‘Body-Left’: ET in the uterine body (CL in left ovary) and group ‘Body-Right’: ET in the uterine body (CL in right ovary). Corpus luteum activity was evaluated in all groups by measuring plasma progesterone concentrations. Embryos were recovered by flushing the uterus on Day 8 after the first mating of the donor and transcervical ET was carried out in recipients 6 days after buserelin administration. Pregnancy rates were: group ‘Left-Ipsilateral’: 50%; group ‘Left-Contralateral’: 20%; group ‘Right-Ipsilateral’: 30%; group ‘Body-Left’ and ‘Body-Right’: 10%. No significant differences ( P = 0.4728) were detected between the pregnancy rates in the five groups. Threading the cervix and transcervical placing of PBS either in the uterine horns or the body did not affect plasma progesterone concentrations in the llama, indicating that the different embryo transfer maneuvers do not interfere with CL viability. To improve pregnancy rates it could be suggested that ET in the left uterine horn with an ipsilateral CL, is the most desirable option.

Half dose of prostaglandin F2a is effective to induce luteolysis in the synchronization of ovulation protocol for fixed-time artificial insemination in buffalo (Bubalus bubalis)

The objective of this study was to evaluate the efficiency of half dose of PGF2alpha in the synchronization of ovulation protocol (GnRH/PGF2alpha/GnRH) for fixed-time artificial insemination in buffalo. Two hundred and sixty two buffaloes (³40 days postpartum) were divided in 2 groups (Group 1, n = 125, GnRH/ ½ dose de PGF2alpha/GnRH; Control Group, n = 137, GnRH/PGF2alpha/GnRH). Animals in Group 1 received 25mg of GnRH IM (Lecirelin, Gestran-plus®) and, seven days later, half dose of PGF2alpha IM (75mg d-cloprostenol, Preloban®). Two days after PGF2alpha injection, all animals received 25mg of GnRH IM (Lecirelin). Animals in Control Group received the same treatment, except for the dose of PGF2alpha(150mg, IM, d-cloprostenol, Preloban®). All animals were fixed-time inseminated 16 hours after the second GnRH injection. Blood samples were collected from 35 buffaloes in Group 1 and 30 buffaloes in Control Group on day of PGF2alpha and second GnRH injections to measure plasma progesterone concentrations. Conception rate was not influenced by PGF2alpha dosage (P>;0.05). Conception rates were 48.00% (60/125) and 40.90% (56/137) in Group 1 and Control Group, respectively. It was observed that 88.60% (n = 35) of the buffaloes in Group 1 and 90.00% (n = 30) in Control Group presented luteolysis 48 hours after the PGF2alpha injection (P>;0.05). In summary, present data show that both treatments result in acceptable conception rates. Therefore, the recommended dose of PGF2alpha can be reduced in 50.00% in the synchronization of ovulation protocol for fixed-time artificial insemination in buffalo.

Reproductive seasonality in female Iberian red deer (Cervus elaphus hispanicus)

This study characterized the seasonal pattern of luteal cyclicity in Iberian red deer (n16), by measuring plasma progesterone concentrations in hinds (female red deer) twice per week from calving (May and June) 1996 until May 1997. In eight of these hinds we also examined plasma prolactin profiles to assess seasonal responses to photoperiod.Plasma progesterone concentration in the 16 hinds studied indicated that the reproductive pattern is seasonal, and lasts for 5.73±0.27 months. After calving, progesterone levels remained basal (no luteal activity) for several months, except in a hind that lost her calf just after calving, and thus did not have to suckle it. This hind showed two consecutive estrus cycles in the month following calving, which suggests that suckling has an inhibiting effect on the resumption of ovarian activity. These results also showed that as long as the hinds do not become pregnant, they show between 5 and 10 estrus cycles per reproductive season (8.06±0.35), ranging between 105 and 249 days from onset of the first cycle to end of the last one. Uninterrupted cycling lasted for 3.5–6.4 months (mean, 4.6±0.24). Cyclic luteal activity was found from October to February in all hinds, with a smaller, but notable proportion in September (56.25%) and March (68.8%), whereas it was negligible in the remaining period. Our results show a reproductive season similar to or longer than that recorded by other authors.Prolactin plasma concentrations showed a yearly trend following that of photoperiod, with peak concentrations from April to July, a decrease in August, minimal concentrations from September to February and a sharp increase in March.

Reproductive and milking performance of a herd of Friesian W N'Dama cattle

Production records of a herd of Friesian W N'Dama cattle reared on-station were examined to assess their potential for use as dairy animals in the tropics. In addition, the radio- immunoassay technique was used to measure plasma progesterone concentrations to determine the attainment of puberty and resumption of ovarian activity postpartum in heifers and cows, respectively. A Friesian bull was always used to serve females on heat. Bull calves were significantly (P (Ghana Journal of Agricultural Science, 1996, 29(2): 115-124)

Advancement of ovulation and oestrus after temporary calf removal and FSH supplementation in postpartum beef cows

To evaluate the effect of temporary calf removal and/or FSH supplementation on post-partum ovarian and oestrous activity, 40 anoestrous multiparous Aberdeen Angus cows under grazing conditions were used. The animals were allocated, on the basis of calving date and body condition, to four initially similar groups according to a 2 × 2 factorial design (ten animals per group) involving a 72 h calf removal (TCR) and exogenous follicle stimulating hormone supplementation (FSH). Anoestrus was determined by measuring plasma progesterone concentrations before treatments, which began at 29.6 ± 4.9 days postpartum. Total FSH supplementation (12 mg, porcine FSH) was performed by six intramuscular decreasing doses (3, 3, 2, 2, 1 and 1 mg, respectively) distributed at 12 h intervals, beginning 24 h before TCR. Oestrus detection (twice daily) and blood sample collection (every 2–3 days) were performed from 7 days before the start of treatments and for 45 days thereafter. The TCR plus FSH group had higher oestrous and ovulation rates at 40 days postpartum (control, 10% and 10%; TCR, 10% and 10%; FSH, 10% and 10%; TCR plus FSH, 60% and 60%) and 60 days postpartum (control, 30% and 30%; TCR, 50% and 50%; FSH, 40% and 50%; TCR plus FSH, 80% and 100%). These differences were significant ( P < 0.05) for oestrous rates at 40 days postpartum and for ovulation rates at 40 and 60 days postpartum. TCR plus FSH significantly shortened ( P < 0.05) the intervals (mean ± s.e.m., days) from calving to first ovulation (control, 61.2 ± 5.5; TCR, 62.8 ± 4.6; FSH, 61.6 ± 3.8 and TCR plus FSH, 43.4 ± 3.1) and a similar trend ( P > 0.05) was observed for the intervals from calving to first oestrus (control, 61.2 ± 5.5; TCR, 62.3 ± 5.4; FSH, 63.4 ± 3.9 and TCR + FSH, 47.7 ± 5.2). The rate of short first luteal phases (⩽ 10 days) was: control, 40%; TCR, 33%; FSH, 43%; TCR plus FSH, 89% ( P < 0.12). In summary, TCR plus FSH resulted in increased ovulation and oestrus rates, shorter postpartum intervals, but subnormal corpora lutea were formed following the first ovulation in early postpartum beef cows.

Ovulation in prenatally androgenized ewes.

The ovarian activity of prenatally androgenized ewes was studied by measuring plasma progesterone concentrations in daily samples of peripheral blood, and by examining the ovaries at laparotomy. Ewes that were exposed to testosterone between days 30 and 80, 50 and 100 or 70 and 120 of foetal life by implanting their mothers with 1 g testosterone, failed to show regular overt oestrous cycles, although some of them ovulated, whereas ewes exposed to testosterone between days 90 and 140 of foetal life had normal oestrous cycles. The incidence of ovulatory failure appeared to increase with age in ewes treated between days 50 and 100 or days 70 and 120 of foetal life.