Pregnancy-specific protein B (PSPB), progesterone and some biochemical attributes concentrations in the fetal fluids and serum and its relationship with fetal and placental characteristics of Iraqi riverine buffalo (Bubalus bubalis).

This study was carried out to demonstrate the pregnancy-specific protein B (PSPB), progesterone and some biochemical parameters concentrations in amniotic fluid, allantoic fluid and fetal serum collected from slaughtered Iraqi riverine pregnant buffaloes at three different months of gestation (6th, 7th and 8th). Ten out of 22 adult buffaloes of 4.6 ± 0.97 years old were used in this study. The buffaloes were mated naturally by monitoring the estrus cycles via appearance of vaginal fluids and mounting by bulls. Pregnancy was checked for these buffaloes by non-returning to estrus for three estrus cycles and assured by rectal palpation on day 61 post-mating (PM). Buffaloes were slaughtered at three different periods of gestation (three at 6th month, four at 7th month and three at 8th month of gestation) to verify the progesterone and PSPB as well as some blood attributes levels (glucose, cholesterol, total protein, albumin, globulins and albumin: globulins ratio) in amniotic fluid (AF), allantoic fluid (LF) and fetal serum (FS). Progesterone was higher (P<0.01) in LF at the 8th month of gestation and lower in FS during the 7th and 8th months of pregnancy. PSPB concentrations were greater in FS (6th and 8th months in particular) than in both AF and LF. The overall mean of cholesterol concentration was higher in FS (P<0.05) followed by AF and LF that had the lowest concentration. The FS exhibited higher total protein during the three gestation periods. Most of fetal and placental measurements increased as the pregnancy advanced. In conclusion, these results described, for the first time, the PSPB and progesterone concentrations and blood characteristics in fetal fluids and serum in water riverine buffaloes during different stages of pregnancy. Progesterone concentrations were greater in allantoic fluid than in other fluids. In contrast, PSPB and other blood attributes were higher in fetal serum than other fluids of Iraqi riverine buffaloes. These findings reflect the changes in hormones, proteins and other metabolites during different gestation periods.

Early pregnancy detection of Iraqi riverine buffalo (Bubalus bubalis) using the BioPRYN enzyme-linked immunosorbent assay for PSPB and the progesterone assay.

This study was undertaken to detect pregnancy in Iraqi riverine buffalo (Bubalus bubalis) using three different methods (rectal palpation, plasma progesterone concentration and detection of the presence of pregnancy-specific protein B (PSPB) with the BioPRYN(®) enzyme-linked immunosorbent assay (ELISA) test. The aim of the study was to identify the most sensitive, early and accurate method for detecting pregnancy. Twenty-two female riverine buffalo that were 6.0 ± 0.93 years old were used. Four blood samples per buffalo were taken via jugular venipuncture at days 22-24, 32-34, 42-44 and 58-61 post-mating (PM) to measure the progesterone concentration (ng/ml) and to detect the presence of plasma PSPB. The rectal palpation method was employed to evaluate all buffalo on days 42-44 and 58-61 PM. The BioPRYN(®) test differed (p<0.01) from the other tests with earlier accuracy for detecting pregnant and non-pregnant buffalo. Eighty-eight percent of pregnant and 76.9% of non-pregnant buffalo were distinguished early (days 22-24 PM) using BioPRYN(®) and plasma PSPB-ELISA level (2.09 ± 0.12 ng/ml) in relation to 66.7% and 53.9% detected using the progesterone assay at similar days (4.30 ± 0.40 ng/ml). In conclusion, these results described, for the first time, the early and accurate pregnancy detection of water riverine buffalo using BioPRYN(®) technology and provided the plasma levels of PSPB using an ELISA test. These findings will improve the reproductive and productive efficiency of Iraqi riverine buffalo by adapting the recent management and reproductive strategies in Iraq and in the world.

Early detection of pregnancy and embryonic loss in dairy cattle by ELISA tests.

Over a 25-month period 8118 blood samples were assayed for the presence of the serum pregnancy specific-protein B (PSPB) and progesterone (P4) concentrations on three Hungarian large-scale dairy farms. Pregnancy (n = 4085) was checked by BioPRYN assay at 30-36 days post-insemination (PI). Samples from all cows that tested not pregnant and from cows with an optical density (OD) reading in the BioPRYN test that was between 0% and 30% above the cutoff OD value were tested for serum P4 concentration. According to serum P4 concentration, cows were assigned to three categories: high (>4 ng/ml), medium (2-4 ng/ml) and low (<2 ng/ml) serum progesterone. The authors predicted a presumed (low) or possible (medium) late embryonic loss (LEL) or maintenance of the pregnancy (high). A total of 710 LELs were detected (17.4%) and 31.8% of them were predicted because of a low OD value at 30-36 days after insemination. Lower PSPB serum level significantly refers for LEL (p < 0.0001). The prediction rate for the true embryonic loss was 31.8% when OD cutoff from 0% to + 30% of cutoff was examined while it was 62.5% when the threshold was OD cutoff of 0% to 10% of cutoff. The authors conclude that BioPRYN was useful for prediction of a part of LEL in dairy cows and serum P4 concentration in these cows related to the rate of LEL.

Effect of mifepristone on pregnancy, pregnancy-specific protein B (PSPB), progesterone, estradiol-17beta, prostaglandin F2alpha (PGF2alpha) and prostaglandin E (PGE) in ovariectomized 90-day pregnant ewes.

The objective of this experiment was to determine the effect of mifepristone, a progesterone receptor antagonist, on pregnancy and secretion of steroids, pregnancy-specific protein B (PSPB) and prostaglandins at mid-pregnancy in ewes. Ninety-day pregnant ewes were ovariectomized (OVX) and treatments were initiated 72 h post-OVX. Ewes received (1) vehicle, (2) prostaglandin F2alpha (PGF2alpha, 8 mg/58 kg/bw, i.m.) 84 h post-OVX, (3) mifepristone (50 mg intrajugular at 72, 84, 96, and 108 h post-OVX), (4) mifepristone (50mg) + PGF2alpha, (5) mifepristone (100 mg intrajugular at 72, 84, 96, and 108 h), and (6) mifepristone (100 mg) + PGF2alpha. Ewes treated with vehicle or PGF2alpha alone did not abort (P > or = 0.05). But, 60, 80, 60, and 100% of ewes treated with mifepristone (50 mg), mifepristone (50 mg) + PGF2alpha, mifepristone (100 mg), and mifepristone (100 mg) + PGF2alpha, respectively, aborted (P < or = 0.05). Profiles of progesterone, estradiol-17beta, prostaglandin E (PGE), or PSPB did not differ (P > or = 0.05) among treatment groups. Profiles of PGF2alpha of treatment groups receiving mifepristone with or without PGF2alpha differed (P < 0.05) from vehicle or PGF2alpha alone-treated ewes. It is concluded that progesterone actions are necessary to suppress uterine/placental secretion of PGF2alpha and that maintenance of critical progesterone: estradiol-17beta and PGE:PGF2alpha ratios are necessary for maintenance of pregnancy.

Effect of the aromatase inhibitor CGS-16949A on pregnancy and secretion of progesterone, estradiol-17beta, prostaglandins E and F2alpha (PGE; PGF2alpha) and pregnancy specific protein B (PSPB) in 90-day ovariectomized pregnant ewes.

The aromatase inhibitor CGS-16949A was used to determine whether CGS-16949A altered secretion of progesterone, estradiol-17beta, PGE (PGE1 + PGE2), PGF2alpha and PSPB. Ninety day pregnant ewes were ovariectomized and received vehicle, PGF2alpha, CGS-16949A or PGF2alpha+CGS-16949A. None of the ewes treated with PGF2alpha, CGS-16949A or PGF2alpha+CGS-16949A aborted (P > or = 0.05) during the 108-h experimental period. Treatment with CGS-16949A lowered (P < or = 0.05) progesterone in jugular venous plasma but concentrations of progesterone were not affected (P > or = 0.05) by treatment with PGF2alpha. Concentrations of estradiol-17beta and PSPB in jugular venous plasma and PGE in inferior vena cava plasma were decreased (P < or = 0.05) by treatment with CGS-16949A. Concentrations of PGF2alpha in inferior vena cava plasma were not affected (P > or = 0.05) by treatment with CGS-16949A. Decreases in estradiol-17beta occurred before decreases in PSPB, which was then followed by decreases in PGE (P < or = 0.05). It is concluded that these data support the hypothesis that estradiol-17beta regulates placental secretion of PSPB; PSPB regulates placental secretion of PGE; and PGE regulates placental secretion of progesterone during mid-pregnancy in ewes.

Effects of GnRH treatment on scrotal surface temperatures in bulls.

Two experiments were conducted to characterize scrotal surface temperature (SST) in bulls treated with gonadotropin releasing hormone (GnRH). In Experiment 1, Angus bulls (n = 10, 18 mo, 597 kg) were given GnRH (400 ng/kg) or saline, IV. Bottom SST increased approximately 1.7 degrees C (P < 0.005) over time (0 to 90 min) at an ambient temperature of 5 degrees C. However, there was no significant effect of GnRH treatment and temperature increases were attributed to stress. When the experiment was repeated at an ambient temperature of 25 degrees C, SST was elevated prior to treatment, with no subsequent significant increase. Experiment 2 was conducted with Charolais bulls (n = 6, 12-14 mo, 517 kg) with an emphasis on minimizing stress. Bottom SST increased approximately 2 degrees C (P < 0.05) between 0 and 45 min after GnRH treatment, supporting the hypothesis that GnRH treatment increases SST in bulls. In conclusion, it was apparent that stress, high ambient temperatures, and GnRH treatment can all increase SST in bulls.

Effect of PGF2alpha, indomethacin, tamoxifen, or estradiol-17beta on incidence of abortion, progesterone, and pregnancy-specific protein B (PSPB) secretion in 88- to 90-day pregnant sheep.

One objective of this experiment was to evaluate our hypotheses that estradiol-17beta regulates secretion of pregnancy specific protein B (PSPB) and that secretion of progesterone during pregnancy is regulated by a prostanoid by examining the effects of prostaglandin F2alpha (PGF2alpha), a luteolyic agent; indomethacin, a prostanoid synthesis inhibitor; tamoxifen, an estrogen receptor antagonist; estradiol 17-beta; and interaction of these factors on the incidence of abortion and progesterone and PSPB secretion. Another objective was to determine if there is a luteal source of PSPB. Weights of corpora lutea were decreased (P < or = 0.05) by PGF2alpha, indomethacin, PGF2alpha + tamoxifen, PGF2alpha + indomethacin, and PGF2alpha + estradiol-17beta but not (P > or = 0.05) by tamoxifen or estradiol-17beta alone. No ewe treated with PGF2alpha alone aborted (P > or = 0.05). Forty percent of ewes treated with PGF2alpha + estradiol-17beta aborted (P < or = 0.05), but ewes were not aborted by any other treatment within the 72-h sampling period. Profiles of progesterone in jugular venous blood differed (P < or = 0.05) among control, indomethacin-, tamoxifen-, and PGF2alpha + indomethacin-treated ewes. Progesterone in jugular venous blood of control ewes decreased (P < or = 0.05) by 24 h, followed by a quadratic increase (P < or = 0.05) from 24 to 62 h. Progesterone in jugular venous blood of indomethacin-, PGF2alpha-, PGF2alpha- + tamoxifen-, PGF2alpha + indomethacin-, PGF2alpha + estradiol-17beta-, and tamoxifen-treated ewes was reduced (P < or = 0.05) by 18 h and did not vary (P > or = 0.05) for the remainder of the 72-h sampling period. Progesterone in vena cava and in uterine venous blood was reduced (P < or = 0.05) at 72 h in PGF2alpha-, indomethacin-, tamoxifen-, PGF2alpha + indomethacin-, PGF2alpha + tamoxifen-, and PGF2alpha + estradiol-17beta-treated ewes. Weights of placentomes did not differ among treatment groups (P > or = 0.05). Profiles of PSPB in inferior vena cava blood differed (P < or = 0.05) among control, estradiol-17beta-, indomethacin-, tamoxifen-, PGF2alpha + indomethacin-, and PGF2alpha + tamoxifen-treated 88- to 90-day pregnant ewes. Concentrations of PSPB in inferior vena cava blood were increased (P < or = 0.05) in indomethacin-, estradiol-17beta-, tamoxifen-, PGF2alpha + tamoxifen-, and PGF2alpha + indomethacin-treated 88- to 90-day pregnant ewes within 6 h and did not vary (P > or = 0.05) for the remainder of the 72-h sampling period. Concentrations of PSPB in uterine venous blood of indomethacin-, tamoxifen-, PGF2alpha + tamoxifen-, and PGF2alpha + indomethacin-treated ewes were greater (P < or = 0.05) at 72 h than at 0 h. PSPB in ovarian venous blood did not differ (P > or = 0.05) adjacent or opposite to the ovary with the corpus luteum. It is concluded from these data that estrogen regulates placental secretion of PSPB and that a prostanoid, presumably prostaglandin E, regulates placental secretion of progesterone during 88-90 days of gestation in sheep and that there is no luteal source of PSPB.

Secretion of progesterone, estradiol-17beta, PGE, PGF2alpha, and pregnancy-specific protein B by 90-day intact and ovariectomized pregnant ewes.

Ninety-day pregnant ewes were either laparotomized, ovaries left in situ or bilaterally ovariectomized, and a jugular venous catheter and an inferior vena cava catheter via the saphenous vein were installed. Seven days later, placenta slices were collected and incubated in vitro for 4 h. Secretions of progesterone, PGE, estradiol-17beta and pregnancy-specific protein B (PSPB) in vitro by placenta from ovariectomized ewes were increased (P < or = 0.05) by 2.7-, 3.6-, 2.2-, and 2.4-fold, respectively, when compared to placenta slices from intact 90-day pregnant ewes. Secretion of PGF2alpha in vitro was unchanged (P > or = 0.05). Ovariectomy decreased (P < or = 0.05) jugular venous progesterone for 78 h followed by a quadratic increase (P < or = 0.05), whereas progesterone remained unchanged (P > or = 0.05) in intact ewes over the 162-h sampling period. Ovariectomy increased (P < or = 0.05) PGE in inferior vena cava plasma over the last half of the 162-h sampling period, whereas concentration of PGF2alpha did not change (P > or = 0.05). Increases in PGE occurred before the increase in progesterone. Concentrations of PSPB in inferior vena cava plasma of ovariectomized pregnant ewes increased (P < or = 0.05) during the last half of the 162-h sampling period, but not in intact ewes (P > or = 0.05). PSPB increased before PGE and progesterone. Concentrations of estradiol-17beta in jugular venous plasma of ovariectomized pregnant ewes increased (P < or = 0.05) during the last half of the sampling period, but not in intact ewes (P > or = 0.05). Increases in estradiol-17beta occurred before increases in PSPB. It is concluded that these data support the hypothesis that estradiol-17beta may control placental secretion of PSPB; PSPB may regulate placental secretion of PGE; and PGE may regulate placental secretion of progesterone.

PGE2 induces its own secretion in vitro by bovine 270-day placenta but not by 200-day placenta.

Two separate experiments were conducted to determine whether prostaglandin (PG) E2 stimulates the secretion of progesterone by 270- or 200-day Brahman placentas in vitro. Secretion of progesterone, PGF2alpha, pregnancy specific protein B, or estradiol-17beta by 270-day Brahman placentas was not affected (p > or = 0.05) by PGE2, during the 4-h incubation period at the doses tested. Indomethacin or meclofenamic acid decreased (p < or = 0.05) 270-day Brahman placental secretion of PGE and PGF2alpha by 98 and 60%, respectively. However, PGE2 induced (p < or = 0.05) its own secretion, but not the secretion of PGF2alpha (p > or = 0.05), by 270-day Brahman placentas, even in the presence of indomethacin or meclofenamic acid at a dose of 100 ng/mL. Also, secretion of 8-Epi-PGE2 by Day 270 Brahman placentas was increased (p < or = 0.05) by PGE2. Secretion of progesterone, estradiol-17beta, or pregnancy specific protein B by 200-day Brahman placentas was not affected by PGE2, 8-Epi-PGE2, PGF2alpha, estradiol-17beta, or trichosanthin during the 4- or 8-h incubation period (p > or = 0.05). Secretion of estradiol-17beta at 8 h was lower (p < or = 0.05) in all treatment groups and did not differ (p > or = 0.05) among the 8-h incubation treatment groups. Secretion of PGE by 200-day Brahman placentas was reduced (p < 0.05) by indomethacin 72 and 82% and by meclofenamic acid 72 and 96%, respectively, at 4 and 8 h when compared to controls. Secretion of PGF2alpha was reduced (p < or = 0.05) 71 and 86% by indomethacin or 89 and 89% by meclofenamic acid at 4 and 8 h, respectively, and did not differ (p > or = 0.05) between 4 and 8 h of incubation. PGE2 did not (p > or = 0.05) induce secretion of PGE above what was added in any treatment group. PGE in culture media was increased (p < or = 0.05) by 8-Epi-PGE2, pregnancy specific protein B, and the 100 ng/mL PGF2alpha dose (p < or = 0.05), but not by PGE2, progesterone, estradiol-17beta, 8-Epi-PGF2alpha, or trichosanthin. Secretion of PGF2alpha by 200-day Brahman placentas was not affected (p > or = 0.05) by 8-Epi-PGE2, progesterone, or estradiol-17beta, but PGF2alpha secretion was increased (p < or = 0.05) by trichosanthin or PGE2, even in the presence of indomethacin or meclofenamic acid. It is concluded that PGE does not affect secretion of progesterone by 200- or 270-day bovine placentas, but, pregnancy specific protein B may regulate placental secretion of PGE. Also, indomethacin and meclofenamic may affect enzymes converting PGH to PGE rather than acting only on cyclooxygenase because indomethacin and meclofenamic acid lowered PGE secretion by 270-day Brahman placentas more than they lowered PGF2alpha. In addition, it is concluded that PGE2 can induce bovine placental secretion of PGE, but this is dependent upon the stage of gestation.

Pregnancy-specific protein B induces release of an alpha chemokine in bovine endometrium.

Pregnancy-specific protein B (PSPB), is secreted from binucleate trophoblast of the bovine conceptus as early as day 15 of pregnancy. The objective of this experiment was to determine if PSPB induced uterine proteins. PSPB was purified from day 120 cotyledons using antibody-based affinity chromatography. Endometrium from day 14 nonpregnant cows (n = 3) was prepared for explant (3H-Leu added) culture. Radiolabeled proteins released into medium were dialyzed, separated using 1D-PAGE, and detected using fluorography and densitometry. PSPB (0, 0.5, 5, 25 & 50 nM) caused a concentration-dependent increase in the release of a radiolabeled 8-kDa uterine protein. Western blots revealed that the 8-kDa protein cross-reacted with antibody against granulocyte chemotactic protein-2 (GCP-2). PSPB also induced release of GCP-2 by bovine endometrial (BEND) cells in primary culture. The induction of GCP-2 by PSPB was blocked by addition of antiserum against PSPB (1:4 molar ratio). This is the first indication that PSPB has a hormonal role in inducing GCP-2, an alpha chemokine that also is induced by interferon-tau during early pregnancy. This chemotactic cytokine may be integral to mediating adhesion, inflammation and angiogenesis associated with early implantation.

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.

Morphologic, endocrine and thermographic measurements of testicles in comparison with semen characteristics in mature Holstein-Friesian breeding bulls.

Twenty Holstein-Friesian breeding bulls (62-79 months of age) were examined 3 times, at 30-day intervals. Scrotal thermograms for assessment of scrotal surface temperature (SST) and blood samples for plasma testosterone concentrations were taken just before and then 45 and 90 min, respectively, after treatment with GnRH (50 micrograms, Gonavet, i.m. per bull). Following GnRH treatment, there generally were significant increases in mean values of both top SST (range, -0.1 to 1.4 degrees C) and bottom SST (range, 0.3 to 1.8 degrees C). Scrotal circumference was highly repeatable but SST and video-measurements of scrotal dimensions were less repeatable, because apparently they were affected by ambient temperature. Plasma testosterone concentrations before GnRH treatment were more repeatable than those after GnRH treatment. Correlations between examinations of 0.67 to 0.81 and -0.14 to 0.47, respectively, but the converse was true for SST measurements. Semen was collected with an artificial vagina 3 times per week for 12 weeks starting 2 weeks before the first examination. The total number of spermatozoa per ejaculate was highly repeatable and the percentage of motile and live spermatozoa were relatively consistent. Separate regressions for each variable and for each examination were conducted for these 3 semen characteristics as dependent variables. For the number of spermatozoa per ejaculate and for the percentage of motile spermatozoa, significant independent variables were plasma testosterone concentrations and difference between top and bottom SST, respectively. The slopes of these equations were nearly all negative and the R2 was from 0.15 to 0.42. For prediction of the percentage of live spermatozoa, both SST gradient and plasma testosterone concentrations were significant independent variables. For these regressions, the slopes were negative and the regression coefficients were generally lower than for the other 2 dependent variables (range, 0.16 to 0.25). Treatment with GnRH and assessment of SST and plasma testosterone concentrations have some correlation with the semen production in the mature bull.

Early pregnancy detection and the hormonal characterization of embryonic-fetal mortality in fallow deer (Dama dama).

The objectives of this investigation were to 1) determine serum concentrations of progesterone (P4), estrone sulfate (E1S) and pregnancy-specific protein B (PSPB) from estrus synchronization through mid-gestation in the fallow doe (Dama dama) and 2) characterize the hormonal profiles of does whose embryos or fetuses died in utero. Ten fallow does were synchronized for 14 d with an intravaginal P4-releasing device (CIDR) and were naturally mated after CIDR removal. Blood samples were collected at CIDR insertion, CIDR removal and at intervals through Day 203 post-CIDR removal for analysis of P4, E1S and PSPB by radioimmunoassay (RIA). Ultrasonography was performed on Days 49 and 69 post-CIDR removal. Serum P4 at the time of CIDR insertion was 4.8 +/- 0.6 ng/ml, and at CIDR withdrawal it was 6.2 +/- 0.3 ng/ml. Concentrations of E1S and PSPB were nondetectable at CIDR insertion. Serum E1S was highest at Day 93, and PSPB was first detectable in pregnant does at Days 27 to 30 post-CIDR withdrawal. Ultrasonography on Day 49 revealed that 6 does were pregnant, 2 were not pregnant and 2 others were diagnosed originally as early pregnant. At Day 69, ultrasonography revealed that 6 does (60%) were pregnant and 4 (40%) were not. A comparison of the ultrasonographic and hormonal data indicated that the 2 does diagnosed as early pregnant on Day 49 had conceived but had lost the pregnancy. A third doe which was pregnant on Day 69 lost the fetus later in gestation. Hormonal profiles of does whose embryo or fetus had died were characterized by erratic P4 and E1S profiles, with PSPB becoming undetectable in the 3 does by Days 49, 65 and 80 post-CIDR removal. These data 1) demonstrate the timing for the collection of serum samples for determining early pregnancy in fallow does using 3 hormonal methods and 2) characterize the hormonal profiles of 3 fallow does with embryonic-fetal loss.

Endocrine and thermal responses to GnRH treatment and prediction of sperm output and viability in holstein-Friesian breeding bulls.

A study was conducted to determine changes in serum LH and testosterone concentrations and in scrotal surface temperature (SST; measured with infrared thermography) following GnRH treatment and to predict the number of spermatozoa collected and the proportion that were viable. Holstein-Friesian breeding bulls (n = 22, average age, 24.3 m.o.; range, 15 to 41 m.o.) were examined twice 30 d apart. Concurrently, semen was collected twice weekly with an artificial vagina. Treatment with GnRH (100 micrograms, i.m.) increased (P < 0.0001) serum LH and testosterone concentrations and increased (P < 0.0001) SST (range 0.6 to 1.1 degrees C; P < 0.05) at the top and bottom of the scrotum. In regression models to predict the total number of spermatozoa, significant independent variables included ultrasonic echotexture of the testes (negative slope), scrotal width (positive slope) and SST at the bottom of the scrotum 45 min after GnRH treatment (positive slope). In regression models to predict the percentage of live spermatozoa, ultrasonic echotexture was a significant independent variable (negative slope). Measurement of testicular ultrasonic echotexture and SST after GnRH treatment augmented measurement of testicular size for predicting the number and percentage of live spermatozoa.

PGE1 or PGE2 not LH regulates secretion of progesterone in vitro by the 88-90 day ovine corpus luteum of pregnancy.

Secretion of progesterone in vitro by mature day 8 ovine corpora lutea (CL) of the estrous cycle was increased linearly by ovine LH (1, 10 and 100 ng/ml) or prostaglandin E2 (PGE2) 1, 10 and 100 ng/ml) in a dose dependent manner (P < or = 0.05). Progesterone secretion in vitro by 88-90 day ovine CL of pregnancy was not affected P > or = 0.05 by LH (1, 10 and 100 ng/ml) while prostaglandin E1 (PGE1) 1, 10 and 100 ng/ml) increased (P < or = 0.05) secretion of progesterone in a dose dependent manner and PGE2 (1, 10 and 100 ng/ml) increased (P < or = 0.05) secretion of progesterone only at the 100 ng/ml dose. Day 8 ovine CL of the estrous cycle did not secrete (P > or = 0.05) detectable quantities of prostaglandin F2 alpha (PGF2 alpha) or prostaglandin E (PGE) while 88-90 day ovine CL of pregnancy secrete PGE (P < or = 0.05) but not PGF2 alpha (P > or = 0.05). Regulation of PGE secretion by 88-90 day ovine CL of pregnancy may be via pregnancy specific protein B (PSPB), which increased (P < or = 0.05) PGE and progesterone but not PGF2 alpha (P > or = 0.05) secretion. Secretion of progesterone by CL of 88-90 days of pregnancy was not affected by IGF1, IGF2, PAF-16, PAF-18, oxytocin, PGI2, PGD2 or leukotriene C4 (P > or = 0.05). It is concluded that PGE1 or PGE2 but not LH regulates secretion of progesterone in vitro by 88-90 day ovine CL of pregnancy. In addition, it is concluded that 88-90 day ovine CL of pregnancy secretes it's own luteotropin, which is PGE. Secretion of PGE by ovine CL of pregnancy may be regulated by PSPB.

Bovine luteal cell production in vitro of prostaglandin E2, oxytocin and progesterone in response to pregnancy-specific protein B and prostaglandin F2 alpha.

A study was conducted to determine the effects of pregnancy-specific protein B (PSPB) and prostaglandin F2 alpha (PGF2 alpha) on bovine luteal cell progesterone, prostaglandin E2 (PGE2) and oxytocin production in vitro. Corpora lutea were enucleated from multiparous cows with normal oestrous cycles during the mid-luteal (days 10-12; n = 5) or late-luteal (days 17-18; n = 5) stage. Mixed large and small cells (1.5 x 10(5) cells per well) were incubated in 500 microliters modified Ham's F-12 medium. Cells were incubated for 18 h before treatments were added. Cells were treated with PSPB (0, 2.5, 5.0 micrograms) and PGF2 alpha (0, 100, 200 ng) in a 3 x 3 factorial arrangement. After treatments were added, media samples were collected at 6 and 12 h. During the 18 h pretreatment incubation, progesterone, PGE2 and oxytocin production was similar between the prospective treatment groups. Progesterone production was greater (P < 0.001) by mid-stage than by late-stage cells. In addition, progesterone decreased (P < 0.001) as incubation time increased. Progesterone production was not affected by PGF2 alpha, but PSPB increased (P < 0.02) progesterone at the 5.0 micrograms dose. Late-stage luteal cells produced more (P < 0.001) PGE2 than did mid-stage cells; PGE2 production decreased (P < 0.001) with increased incubation time. Luteal PGE2 production increased in response to PSPB treatment (P < 0.01) and PGF2 alpha treatment (P < 0.001). Luteal oxytocin production was greater (P < 0.01) by mid-stage compared with late-stage cells. Oxytocin production decreased (P < 0.001) with incubation time in mid-stage cells, but in late-stage cells oxytocin production was similar over time. Neither PSPB nor PGF2 alpha had an effect on oxytocin. These results indicate that PSPB does not affect luteal oxytocin, but does increase progesterone and PGE2 production. In addition, PGF2 alpha increases luteal PGE2, but does not affect progesterone or oxytocin production. These data do not show an interaction between PSPB and PGF2 alpha in regulating bovine luteal cell endocrine function.

Effect of histone-H2A (H-H2A), platelet activating factor (PAF) and pregnancy specific protein B (PSPB) on secretion of prostaglandins E and F2 alpha (PGE; PGF2 alpha) by bovine endometrium and H-H2A on basal secretion of luteinizing hormone (LH) by bovine pituitary cells in vitro.

Effects of H-H2A, PSPB or PAF on day 16 bovine endometrial secretion of PGE and PGF2 alpha and H-H2A on basal secretion of LH by bovine pituitary cells in vitro were examined in two experiments. PAF (P < or = 0.08) and H-H2a + PAF (P < or = 0.10) treatment for two hours in an in vitro perfusion system tended to increase secretion of PGF2 alpha expressed as a proportion of the prechallenge concentrations of PGF2 alpha by day 16 bovine caruncular endometrium, which occurred during the two-hour period after treatment removal. PGF2 alpha was increased (P < or = 0.02) in the two hour period after the treatment was removed in both control and H-H2A-treated endometrium. H-H2A (P < or = 0.07) and PSPB (P < or = 0.08) tended to increase PGE, while H-H2A + PSPB (P < or = 0.05) and H-H2A + PAF (P < or = 0.03) increased secretion of PGE expressed as a proportion of the prechallenge concentrations of PGE by day 16 bovine caruncular endometrium during the challenge and postchallenge treatment periods, but did not differ (P < or = 0.05) between the two-hour challenge period and the two-hour postchallenge period after removal of the treatment. In experiment two, H-H2A decreased (P < or = 0.05) basal secretion of LH by bovine pituitary cells in vitro. These data suggest that H-H2A and PSPB preferentially stimulate secretion of PGE by bovine endometrial tissue and may play a role in maternal recognition of pregnancy, while PAF increased PGF2 alpha secretion. In addition, H-H2A may play a role in regulating secretion of LH by the pituitary. Key words: Pregnancy Specific Protein B, Histone-H2A, Prostaglandin E and F2 alpha, Platelet Activating Factor, Endometrium, Cow.

Effect of pregnancy-specific protein B on luteal cell progesterone, prostaglandin, and oxytocin production during two stages of the bovine estrous cycle.

This experiment was designed to evaluate the effects of pregnancy-specific protein B (PSPB) on luteal cell progesterone (P4), PGF2 alpha, PGE2, and oxytocin secretion. Corpora lutea were collected during the mid (d 10 to 12; n = 5) or late luteal (d 17 to 18; n = 5) stage of the estrous cycle. Large and small cells (1.5 x 10(5)/well) were treated with PSPB (0, 2.5, or 5.0 micrograms) and LH (0, 50, or 100 ng) in a 3 x 3 factorial arrangement. Cells were incubated for 18 h before adding treatments; after treatments, medium was collected at 6 and 12 h. During the 18-h pretreatment period, P4, PGF2 alpha, PGE2, and oxytocin production was similar between the prospective treatment groups. The PSPB did not affect P4 production. Stage of the cycle (stage) x time interaction (P < .001) indicated that mid-stage luteal cells produced more P4 than late-stage cells; regardless of stage, P4 decreased with time. The time x LH interaction (P < .001) revealed that at 6 and 12 h the 50- and 100-ng doses of LH increased P4 to greater than the 0-ng dose. Production of PGF2 alpha by mid-stage cells was similar among the three PSPB treatments; however, PGF2 alpha production by late-stage cells increased (P < .01) in response to the 5.0-micrograms dose of PSPB. The LH did not affect PGF2 alpha production. Late-stage luteal cells produced more (P < .001) PGF2 alpha than mid-stage cells during the 18-h pretreatment period and at 6, but not 12, h.(ABSTRACT TRUNCATED AT 250 WORDS)

Prostaglandin F2 alpha, progesterone and oxytocin production by cultured bovine luteal cells treated with prostaglandin E2 and pregnancy-specific protein B.

The objectives of this experiment were to study the effects of pregnancy-specific protein B (PSPB) and prostaglandin E2 (PGE2) on bovine luteal cell progesterone (P4), prostaglandin F2 alpha (PGF2 alpha) and oxytocin production. Corpora lutea were collected during the mid- (days 10-12; n = 5) or late-luteal (days 17-18; n = 5) stages of the estrous cycle. Luteal cells were dispersed and accessory cells removed. Luteal cells (1.5 x 10(5)) were incubated in a 3 x 3 factorial arrangement and treated with PSPB (0, 2.5, or 5.0 micrograms) and PGE2 (0, 100, or 200 ng) in 500 microL of Ham's F-12 medium. All cells were incubated for 18 h before adding treatments. Samples were then collected at 6 h and 12 h. During the 18 h pretreatment period, P4, PGF2 alpha, and oxytocin production was similar between the prospective treatment groups. The PSPB failed to increase P4 production. The PGE2 x time interaction showed that P4 increased in response to PGE2 treatment at 6 h (P < 0.001) and 12 h (P < 0.03). Also, the stage x time interaction indicated that mid-stage cells produced more (P < .001) P4 than late-stage cells during the pretreatment period at 6 h and 12 h. The PSPB did not alter PGF2 alpha production by mid-stage cells, but increased (P < .05) PGF2 alpha by late-stage cells. Also, PGE2 stimulated (P < 0.001) PGF2 alpha secretion by both mid- and late-stage cells; luteal cells treated with 200 ng of PGE2 produced more (P < 0.001) PGF2 alpha than 100 ng of PGE2. Oxytocin secretion was not changed by treatment with PGE2 or PSPB. Oxytocin production was greater (P < 0.001) by mid-stage than late-stage cells during the pretreatment period at 6 h and 12 h. Oxytocin production was similar between the 6 h and 12 h culture times within stage of the cycle. These data indicate that PSPB does not change bovine luteal cell P4 or oxytocin production, but elevates PGF2 alpha in late-stage cells. The PGE2 increases both P4 and PGF2 alpha, but does not alter oxytocin production. Lastly, PSPB and PGE2 do not interact to promote P4 PGF2 alpha, or oxytocin production by cultured bovine luteal cells.

Fertility of bull semen packaged in .25- and .5-milliliter french straws.

The fertility of bull semen packaged in .25- and .5-mL french straws was compared. One ejaculate from each of five Holstein bulls was split, extended to 10 x 10(6) spermatozoa/inseminate dose in whole homogenized milk, packaged in .25- and .5-mL french straws, frozen in liquid nitrogen (LN) vapor, and stored in LN. Semen was thawed at 37 degrees C for 30 s. Synchronized heifers (n = 1,360) were inseminated (during a 12-mo period) with semen packaged in either a .25- or .5-mL french straw. Blood was collected on the day of insemination and the serum was assayed for progesterone. Heifers with blood progesterone levels of > 1 ng/mL were eliminated from the data. Blood was collected at 30 to 45 d after insemination and the serum was assayed for the presence of bovine pregnancy-specific protein B (bPSPB) by RIA to determine pregnancy. Conception was 63.6 and 62.0% (P = .55) for semen packaged in the .25- and .5-mL french straws, respectively. There was neither a bull x packaging unit interaction (P = .49) nor a day of insemination x packaging unit interaction (P = .87). Conception among bulls ranged from 57.1 to 68.0% (P = .19). No evidence was found that meteorological factors influenced conception. Under the conditions of this experiment, semen packaged in the .25- and .5-mL french straw had similar fertility.