R.D. Randel

Seasonal effects on female reproductive functions in the bovine (Indian breeds)

Abstract Reproductive function is mediated by season in the Indian breeds of cattle (Bosindicus). The reproductive endocrinology of Bosindicus cattle differs from that of Bostaurus breeds; the estrus is shorter and less intense and occurs late in relation to an estrogen stimulus. Moreover, the Bosindicus female has a smaller preovulatory surge of luteinizing hormone (LH), which occurs earlier relative to the onset of estrus, and she ovulates sooner after the onset of estrus. The corpus luteum is smaller and contains less progesterone, and the serum progesterone concentration is lower in Bosindicus females. Furthermore, they have fewer preovulatory LH surges than Bostaurus females and their luteal cells are less responsive to LH in vitro during the winter. Their fertility is lower during the late fall and winter months. For Bosindicus cattle, recovery of transferable embryos and survival of embryos in the recipient are at their maximum from July through October.

Effect of luteinizing hormone (LH), PGE2, 8-EPI-PGE1, 8-EPI-PGE2, trichosanthin, and pregnancy specific protein B (PSPB) on secretion of progesterone in vitro by corpora lutea (CL) from nonpregnant and pregnant cows.

Secretion of progesterone by Day 14 bovine corpora lutea (CL) of the estrous cycle and Day 200 CL of pregnancy was evaluated in vitro to determine what regulates secretion of progesterone by CL of pregnancy. Weights of Day 200 CL of pregnancy (4356 +/- 223 g) were heavier when compared to Day 14 CL of the estrous cycle of Brahman cows (3643 +/- 128 g; p < or = 0.05); however, both Day 14 and Day 200 minced CL slices secreted similar basal amounts of progesterone per unit mass (p > or = 0.05). Secretion of progesterone in vitro by Day 14 CL of the estrous cycle was increased at 4 and 8 h (p < or = 0.05) by 10 or 100 ng/mL luteinizing hormone (LH) and did not differ between doses (p > or = 0.05). Progesterone secretion in vitro by Day 200 CL of pregnancy was not increased (p > or = 0.05) by LH at 4 or 8 h. However, progesterone secretion in vitro by Day 14 CL of the estrous cycle or Day 200 CL of pregnancy was increased (p < or = 0.05) at 4 h by 10 or 100 ng/mL PGE2, which did not differ by dose or reproductive status (p > or = 0.05). At 8 h, Day 14 CL of the estrous cycle secretion of progesterone in vitro was increased (p < or = 0.05) by both doses of PGE2 but only at 8 h by 100 ng/mL from Day 200 CL of pregnancy (p < or = 0.05). Secretion of progesterone in vitro was not affected (p > or = 0.05) by 10 or 100 ng/mL 8-Epi-PGE1 or 8-Epi-PGE2 at 4 or 8 h from Day 14 CL of the estrous cycle or Day 200 of pregnancy. Trichosanthin increased (p < or = 0.05) secretion of progesterone in vitro by 10 ng/mL at 4 h and at 8 h by Day 14 CL of the estrous cycle or at 8 h by Day 200 CL of pregnancy but trichosanthin at 100 ng/mL did not affect (p > or = 0.05) secretion of progesterone in vitro by Day 14 CL of the estrous cycle or Day 200 CL of pregnancy at 4 or 8 h. Pregnancy specific protein B (PSPB) increased (p < or = 0.05) secretion of progesterone in vitro at 4 and 8 h by Day 14 CL of the estrous cycle and did not differ between incubation times (p > or = 0.05). PSPB increased secretion of progesterone at 4 h but not at 8 h (p > or = 0.05) by Day 200 CL of pregnancy. These data suggest that PGE2 or PSPB but not LH, 8-Epi-PGE1 or 8-Epi-PGE2 regulates luteal secretion of progesterone by bovine CL at mid-pregnancy. In addition, it is suggested that weights of bovine CL of pregnancy increase to compensate for a lack of placental secretion of progesterone.

Effects of Luteinizing Hormone (LH), PGE2, 8-Epi-PGE1, 8-Epi-PGF2α, Trichosanthin and Pregnancy Specific Protein B (PSPB) on Secretion of Prostaglandin (PG) E (PGE) or F2α (PGF2α) In Vitro by Corpora Lutea (CL) From Nonpregnant and Pregnant Cows

Abstract Both Day 14 corpora lutea (CL) of the estrous cycle and Day 200 CL of pregnancy secrete detectable prostaglandin E (PGE) and prostaglandin F 2 α (PGF 2 α) in vitro . Corpora lutea from Day 200 pregnant cows secrete more PGE and PGFα in vitro than Day 14 CL of the estrous cycle when incubated in control medium without treatments ( p ≤ 0.05). In addition, secretion of both PGE and PGF 2 α in vitro by both Day 200 CL of pregnancy and Day 14 of the estrous cycle increase ( p ≤ 0.05) with time in culture in the absence of treatments. The PGE: PGF 2 α ratio secreted at 4 h in the absence of treatments by Day 14 CL of the estrous cycle was 1.2 and at 8 h was 1.0 and did not differ ( p ≥ 0.05), while the PGE: PGF 2 α ratio secreted by 200 day CL of pregnancy in the absence of treatments at 4 h was 0.8 and at 8 h decreased ( p ≤ 0.05) to 0.4. The PGE: PGF 2 α ratio at 8 h by 200 day CL of pregnancy was lower ( p ≤ 0.05) than in the Day 14 CL of the estrous cycle at 4 or 8 h. Secretion of PGE or PGF 2 α was affected by luteinizing hormone, PGE 2 , 8-Epi-PGE 1 , 8-Epi-PGE 2 , trichosanthin, and pregnancy specific protein B (PSPB) and was time and dose dependent ( p ≤ 0.05). In summary, the altered ratio of PGE: PGF 2 α may explain the decreased secretion of progesterone at 8 h by Day 200 CL of pregnancy reported previously from the same samples. In addition, caution should be exercised in interpretation of progesterone secretion data with bovine CL studies in vitro . Also, PSPB may play an indirect role through PGE to regulate bovine luteal secretion of progesterone.

Effects of naloxone and animal temperament on serum luteinizing hormone and cortisol concentrations in seasonally anestrous Brahman heifers

The effect of endogenous opioid peptides (EOP) and individual animal temperament on serum luteinizing hormone (LH) were investigated in seasonally anestrous Brahman heifers (n = 24). Animals that had shown behavioral estrus in previous months but that had not returned to estrus for at least 30 d were selected. The heifers were ranked by temperament (tame = 1, normal = 2, wild = 3) and randomly allotted into three groups. Blood was collected from one heifer of each group per day. Blood samples were taken via jugular cannula every 15 min for 6 h and every 30 min for another 4 h. After the first hour of sampling, the heifers received intravenous saline (SAL, n = 8); naloxone (LN, 0.5 mg/kg i.v., n = 8); or naloxone (HN, 1.0 mg/kg i.v., n = 8). Three hours after naloxone treatment, each heifer was given gonadotropin releasing hormone (GnRH, 100 microg i.m.). All samples were processed to yield serum and were assayed for LH by radioimmunoassay (RIA). Hourly samples were assayed for cortisol by RIA. The area under the LH curve 60 min postnaloxone treatment was higher in LN and HN than in SAL (57.0 and 40.8 vs 6.1 units; P<0.01); and the area under the 180 min postnaloxone curve remained higher in LN than in SAL (106.2 vs 35.1 units; P<0.05). Cortisol concentrations 60 min postnaloxone administration were above prenaloxone levels(38.2 vs 26.7 ng/ml; P<0.0002). Temperament scores of heifers were positively correlated with cortisol release. The area under the cortisol curve had a negative correlation with mean LH. Serum LH concentrations appear to be suppressed by EOP in seasonally anestrous Brahman heifers, and EOP appear to reduce serum cortisol concentrations. Excitable heifers had higher concentrations of serum cortisol, which negatively affected serum LH concentrations.

Successful superovulation, nonsurgical collection and transfer of embryos from Brahman cows.

Nonsurgical recoveries and transfers of embryos were performed at the McKellar Embryo Transplant Center from 122 superovulated Brahman cows. FSH-P (Armour) was used to superovulate all cows at dose levels ranging from 36 to 48 mg total FSH-P. Luteal regression was induced by use of 40 mg PGF(2(alpha)) in all 122 cows. Embryos were transferred into recipients 6, 7 or 8 days after observed estrus. Embryos were successfully collected from 82% of the FSH-P treated cows. The dose level of FSH-P affected numbers of embryos collected (P<.05). Numbers of embryos collected from cows superovulated with 36-38, 40, 42, 43, 44, 45, 46 and 47-48 mg FSH-P were 2.8 +/- 1.0, 6.8 +/- 1.1, 9.4 +/- 1.4, 10.0 +/- 2.7, 7.1 +/- 1.6, 6.8 +/- 2.0, 5.0 +/- 1.7 and 4.6 +/- 2.0 embryos, respectively. The dose level of FSH-P also affected numbers of embryos transferred (P<.10). Number of embryos transferred from cows superovulated with 36-38, 40, 42, 43, 44, 45, 46 and 47-48 mg FSH-P were 2.8 +/- 1.9, 5.2 +/- 0.9, 6.9 +/- 1.2, 6.7 +/- 2.1, 4.8 +/- 1.3, 5.1 +/- 1.4, 3.4 +/- 1.2 and 3.2 +/- 2.1 embryos, respectively. The developmental stage (D) of the embryo was also a factor in pregnancy rate of recipients (morula = 13.8%, blastocyst = 22.1% and expanded blastocyst = 29.9%; P<.005). The skill of the technician (T) transferring the embryo had a dramatic effect upon subsequent pregnancy rate of the recipients (T 1 = 46.0% vs T 2 = 22.6% pregnancy rate; P<.005). Pregnancy rate of recipients was also affected by the stage postestrus (S) at which the embryo was transferred (day 6 = 23.5%, day 7 = 25.5% and day 8 = 42.3% pregnancy rate; P<.05). Interactions were found between T x S, T x D, S x D and T x S x D (P<.05). These data indicate that use of 40, 42, or 43 mg total doses of FSH-P were quite effective in superovulating the Brahman cow. Recipients transferred on day 8 postestrus achieved higher pregnancy rates than recipients transferred on days 6 or 7 postestrus. Embryos transferred in the expanded blastocyst stage of development proved to yield the highest pregnancy rates in recipients.

ARTIFICIAL INSEMINATION, HYBRIDIZATION AND PREGNANCY DETECTION IN SIKA DEER (Cervus nippon)

Artificial insemination (AI) was performed on sika hinds (Cervus nippon ) receiving various dosages of pregnant mare serum gonadotropin (PMSG; Year 1: 0, 50 and 100 IU; Year 2: 100 and 150 IU) and using semen collected from elk and 1 2 elk x 1 2 sika stags. The time from synchronization device removal (CIDR vs norgestomet) to estrus was determined through observations of mounting activity. Methods for pregnancy detection, serum progesterone (P4), estrone sulfate (E1S), pregnancy-specific protein B (PSPB) and ultrasonography, following AI (Year 1: AI, Days 28 and 48 after AI; Year 2: AI, Days 42, 53 and 100 after AI) and a 90-d natural breeding season were investigated. From available production data, body weights were compared among sika and 1 4 elk x 3 4 sika hybrids relative to age. Pregnancy rates tended (P < 0.10) to differ relative to PMSG treatment and sire; administration of 0 IU PMSG resulted in fewer hinds becoming pregnant to AI than 50 or 100 IU of PMSG. Hinds receiving 100 IU of PMSG had higher (P < 0.05) pregnancy rates than hinds receiving 150 IU PMSG. Time to standing estrus did not differ (P > 0.10) between the CIDR and norgestomet groups. Pregnancy rates 50 d after a 90-d breeding season were similar (P > 0.10) between ultrasound (70.9%) and PSPB (61.6%). Serum P4 after 90 d in breeding groups and 50 d after stag removal were higher (P < 0.05) for pregnant than open hinds. Pregnancy rates (Year 1) 48 d after AI were similar (P > 0.10) between ultrasound (49.0%) and PSPB (37.3%). Serum P4 28 and 48 d after AI were higher (P < 0.05) for pregnant than open hinds. Serum E1S was higher (P < 0.01) for pregnant than open hinds 48 d after AI. Pregnancy rates (Year 2) 100 d after AI did not differ (P > 0.10) between ultrasound and PSPB (66.7%). Serum P4 was higher (P < 0.03) in pregnant than open hinds at 42, 53 and 100 d after AI. At 100 d after AI, pregnant hinds had higher (P < 0.002) serum E1S than open hinds. At 6 to 8 and 11 to 13 mo of age, 1 4 elk x 3 4 sika males tended (P < 0.08) to be heavier than sika males, while 1 4 elk x 3 4 sika females were heavier (P < 0.05) than sika females at all ages. In summary, this study documents the use of AI and methods for pregnancy detection in sika hinds as well as preliminary information regarding the production of elk-x-sika hybrids.

Methods for pregnancy determination and the effects of body condition on pregnancy status in Rocky mountain elk ( Cervus elephus nelsoni )

The objective of this study was to determine the effectiveness of transrectal ultrasonography and serum progesterone (P(4)), estrone sulfate (E(1)S) and pregnancy-specific protein B (PSPB), without prior knowledge of reproductive status, in detecting pregnancy in elk cows. In addition, body weight and body condition score (BCS) were determined to assess whether body condition affects pregnancy status in elk cows. Twenty-five elk cows were sampled during the early rut (Period 1) and after the rut (Period 2), an interval of 120 d. Age, weight, BCS and blood samples, for P(4), E(1)S and PSPB determinations, were taken at Periods 1 and 2. Ultrasonography was performed at Period 2. The younger elk cows weighed less (P<0.05) than older cows. However, pregnancy status was not affected (P> 0.10) by age or weight of the cow. Elk cows that calved had higher (P<0.02) BCS at Periods 1 and 2 than cows that remained open. Serum P(4) and E(1)S were higher (P<0.0001) in pregnant cows at Period 2 than in open cows. Progesterone was 85.8% accurate in detecting pregnant versus open cows at Period 1, while E(1)S and PSPB were not effective. Elk cows at Period 1 were <20 d pregnant with the exception of 1 cow at 46 d. Ultrasonography was 92% accurate, P(4) was 95% accurate, and E(1)S and PSPB were both 100% accurate in determining pregnant versus open cows at Period 2. Pregnant cows at Period 2 were all > 100 d pregnant. Ultrasonography, serum E(1)S and PSPB all may provide a reliable means for pregnancy diagnosis in elk cows at > 100 d of gestation, while serum P(4) may be effective when multiple samples are compared during or after the rut, or when used in combination with the other diagnostic methods described. Further research is needed to determine the optimum time period after breeding in elk cows for accurate pregnancy detection through hormonal analysis.

Exogenous PGF2α enhanced GnRH-induced LH release in postpartum cows

This study evaluated the effect of exogenous PGF(2)alpha on circulating LH concentrations in postpartum multiparous (n = 32) and primiparous (n = 46) Brahman cows. The cows were randomly allotted within parity and calving date to receive 0, 1, 2 or 3 mg im PGF(2)alpha (alfaprostol)/100 kg body weight (BW), with or without GnRH on Day 30 after calving. Blood samples were collected at weekly intervals from calving through treatment. Serum progesterone concentrations were determined using RIA procedures to assure that only anestrous cows were treated. Sterile marker bulls were maintained with cows on Coastal bermudagrass pastures until the first estrus was detected. Multiparous cows had a shorter (P < 0.05) interval from calving to estrus than did primiparous cows. Serum LH was affected by time (P < 0.0001), PGF(2)alpha dose (P < 0.0002), GnRH (P < 0.0001), parity by PGF(2)alpha dose (P < 0.0003), PGF(2)alpha dose by GnRH (P < 0.0009), parity by GnRH (P < 0.0008), and by parity by PGF(2)alpha dose by GnRH (P < 0.0005). Multiparous cows not receiving GnRH had higher mean serum LH (P < 0.02), LH peak pulse height (P < 0.03), and area under the LH release curve (P < 0.03) compared with primiparous cows. The number of LH pulses/6 h was greater (P < 0.06) in multiparous than primiparous cows, and was greater (P < 0.02) in multiparous cows receiving 3 mg/100 kg BW than in cows receiving 2 mg/100 kg BW, but not in the controls or in cows receiving 1 mg/100 kg BW. Exogenous GnRH resulted in increased (P < 0.0001) serum LH concentrations in all cows, and LH was enhanced (P < 0.0009) by simultaneous treatment with PGF(2)alpha. Primiparous cows had a greater response (P < 0.0005) to PGF(2)alpha and GnRH compared with multiparous cows. Pituitary release of LH in response to GnRH was enhanced by simultaneous exposure to PGF(2)alpha in Day 30 postpartum cows.

Reproductive studies of Brahman cattle II. Luteinizing hormone patterns in ovariectomized Brahman and Hereford cows before and after injection of gonadotropin releasing hormone

Abstract Six Brahman and six Hereford long-term ovariectomized cows were bled via tail vessel at 15 minute intervals for a period of 4 hours. Serum was collected and analyzed via radioimmunoassay (RIA) for luteinizing hormone (LH) to determine if ovariectomized Brahman and Hereford cows have pulsatile LH patterns and if breed of animal influenced LH patterns. Brahman and Hereford ovariectomized cattle did have pulsatile LH patterns. Although the trend was for higher LH levels in Hereford than Brahman cows there were no significant differences in mean serum LH levels, number or magnitude of LH peaks or serum LH pulse height. Six Brahman and five Hereford long-term ovariectomized cows were injected (IM) with a single dose of 500μg of gonadotropin releasing hormone (GnRH). Animals were bled via tail vessel at 15 minute intervals for a period of 6 hours. Serum was assayed for LH via RIA to determine if ovariectomized Brahman and Hereford cows differ in GnRH induced LH response. All animals showed increased serum LH in response to GnRH injection within the first 15 minute collection interval. There were no significant differences in duration of response between ovariectomized Brahman or Hereford cows. Ovariectomized Brahman cows had significantly lower (P

Reproductive studies of Brahman cattle IV. Luteinizing hormone levels in ovariectomized Brahman, Brahman × Hereford and Hereford cows following a 20 mg dose of Estradiol-17β☆☆☆★

Luteinizing Hormone (LH) levels were quantitated by radioimmunoassay (RIA) in six mature, long-term ovariectomized cows each of Brahman (B), Brahman × Hereford (B×H) and Hereford (H) breeding following an in-tramuscular injection of 20 mg of Estradiol-17β (E) suspended in corn oil. After E administration all cows were bled via coccygeal venipuncture every two hours from 0–8 hours post-injection, every hour from 9–24 hours post-injection, concluding with bleedings every two hours from 26–36 hours post-injection. An LH surge was observed in 56B cows, 66 B×H cows and 66H cows. Basal LH levels (mean of first eight data points of each breed type) did not differ (P>.10) between B (3.5 ng/ml), B×H (2.4 ng/ml) and H (2.4 ng/ml). Elapsed time from E injection to peak LH value varied significantly (P<.05) between B, B×H and H, respectively (27.8 hrs, 23.8 hrs, 22.2 hrs). Peak LH values also varied between breed (B, 20.2 ng/ml; B×H, 36.0 ng/ml; H, 113.2 ng/ml: P<.005). The area under the LH curve differed significantly between B, B×H and H (P<.05), however, the duration of the LH surge was not different between breeds; B (13.2 hrs), B×H (16.2 hrs) and H (15.3 hrs). Overall significant period effects (P<.05), breed effects (P<.10) and period × breed interactions (P<.05) were found. In summary, B are less reactive to a 20 mg dose of E than are B×H or H using the following criteria: time to peak LH value, peak LH value and area under the LH curve. These data strongly indicate inherent differences between breeds regarding estrogen feedback mechanisms at the hypophysial-hypothalamic axis.