Detection of pregnancy by radioimmunoassay of a novel pregnancy-specific protein in serum of cows and a profile of serum concentrations during gestation

The development of a double antibody radioimmunoassay for a bovine pregnancy-specific protein (pregnancy-specific protein B; PSPB) is presented. By means of this assay, PSPB could be measured in serum of pregnant cows. Five dairy cows were bled throughout gestation to measure serum levels of PSPB. Serum concentrations (means +/- SE) exceeded 1 ng/ml by 30 days postbreeding and increased gradually through three months (9 +/- 0.6 ng/ml), six months (35 +/- 6 ng/ml), and nine months (150 +/- 75 ng/ml) of gestation. Maximum levels of PSPB (542 +/- 144 ng/ml) were reached two days before parturition and then steadily declined to less than 78 ng/ml by 21 days postpartum. In 21 cows bled daily from 15 through 30 days postbreeding, PSPB could be measured in a few cows before and in most cows by 24 days after breeding. In a commercial herd of 102 beef cows, the assay could detect pregnancy earlier and more accurately than the routine method of rectal palpation. This radioimmunoassay measures a unique antigen that, for the first time, provides a serological method for detecting pregnancy in cows.     

Use of milk progesterone enzyme immunoassay for early pregnancy diagnosis in cows

An enzyme immunoassay using beta-galactosidase as the tracer was applied to determine milk progesterone level in cows. The novel method was reliable and practicable and required only a spectrophotometer and a centrifuge as major equipment. The milk progesterone enzyme immunoassay successfully diagnosed early pregnancy in cows. Milk samples were collected from 268 Holstein-Friesian cows in commercial dairy herds on 20, 21 or 22 days(usually 21 days) after insemination. Progesterone level in skim milk higher than 1.0 ng/ml indi-cated pregnancy. Pregnancy was confirmed by rectal palpation on 60 to 120 days after insemination. The accuracy of the milk progesterone test was 60.0 % (132 220 ) for a positive diagnosis and 100 % (48 48 ) for a negative result. A high incidence of embryonic death, 27.9 % (51 183 ), may have reduced accuracy for a positive test result. The enzyme immunoassay may be an alternative to radioimmunoassay in milk progesterone analysis for pregnancy diagnosis.     

Comparison of ovarian palpation, milk progesterone and plasma progesterone in the cow

Holstein cows were examined twice weekly, beginning at 13 to 18 days after parturition, using palpation per rectum and milk progesterone (P4) assay to determine the functional status of the corpus luteum (CL). These results were then compared with P4 concentrations in the plasma to determine which test was the more accurate in detecting functional luteal tissue, or if both tests were needed for optimal accuracy. The tests were found to be comparable except when the plasma P4 concentration was <1. 0 ng/ml. At this level, errors due to palpation occurred more frequently than those due to milk P4 concentrations since the still palpable CL of pregnancy could be mistaken for a functional CL at 14 to 21 days after parturition. However, at all the other concentrations including when plasma P4 was <1.0 ng/ml and the cows were more than 21 days post partum, neither the milk P4 assay nor palpation per rectum could be considered the better indicator of luteal status. Therefore, we conclude that the combined use of both tests does not afford better assessment of luteal function than either test alone.     

Fetal survival in the cow after pregnancy diagnosis by palpation per rectum

It is desirable to determine wheter a cow has failed to become pregnant as early as possible, preferably prior to 50 d after insemination. Although palpation per rectum has been the generally accepted method of pregnancy diagnosis in cattle, the procedure may be a significant iatrogenic cause of fetal attrition. In a study conducted at a North Florida dairy from January through June 1982, pregnancy was determined in 192 Holstein-Friesian cows by measuring low milk progesterone (P4) content on day of insemination (Day 0) and elevated P4 on Days 21 and 24. Pregnant cows were randomly assigned to a treatment and a control group. Cows in the treatment group (n = 85) were palpated per rectum twice between Days 42 and 46 after insemination. Cows in the control group (n = 107) were not palpated until both groups were palpated at Day 90. Palpation, done by two experienced clinicians, consisted of palpation of fetal fluid fluctuation, identification of the amniotic vesicle, and slipping of the chorioallantoic membranes. In both groups of cows fetal viability was monitored by milk P4 content. Last milk (5 to 15 ml) was collected from one front quarter on Days 0, 21, and 24 and twice weekly thereafter through Day 63. Milk was defatted by centrifugation and the fat-free milk progesterone content measured by a radioimmunoassay without extraction. The milk P4 test was 80.0% accurate in determining pregnancy in the palpated and nonpalpated cows. In the cows palpated on Days 42 to 46, pregnancy rates declined by 7.5% as determined by palpation at Day 90, or by 11.4% as determined by milk P4 content through Day 63 (both values P < 0.05). Cows that were not palpated on Days 42 to 46 showed a 1.9% increase or a 4.3% decline in pregnancy rates as determined by the same criteria. Before palpation, at Days 42 to 46, pregnancy rates were better in cows that were inseminated in winter (January to March) than in spring (April to June) (82.3% vs 61.6%; P < 0.05); P4 content was higher (winter > spring = 2.13 ng/ml vs 1.38 ng/ml; P < 0.05). First-lactation cows had higher P4 values on Days 21 and 24 than older cows (P < 0.01).     

Progesterone supplementation during the early fetal period reduces pregnancy loss in high-yielding dairy cattle

It was hypothesized that sub-optimal progesterone concentrations during the late embryo and early fetal period may act to compromise conceptus development in dairy cattle. The aim of the present study was to test this hypothesis by supplementing pregnant cows with exogenous progesterone following pregnancy diagnosis. The study population consisted of 1098 pregnant lactating cows. Pregnancy was diagnosed by transrectal ultrasonography between 36 and 42 days after insemination. Animals found to be pregnant were randomly assigned to the Control (untreated cows, n = 549) or Treatment (n = 549) groups. Cows in group Treatment were fitted at pregnancy diagnosis with a progesterone releasing intravaginal device (PRID) containing 1.55 g of progesterone, for 28 days. Cows were then subjected to a further diagnosis by palpation per rectum on Day 90 of gestation. Pregnancy loss was registered in 95 (8.7%) cows on Day 90 of pregnancy: 66 (12%) in group Control and 29 (5.3%) in group Treatment. Logistic regression analysis indicated that there were no significant effects of herd, bull, milk production, service number, days in milk at pregnancy and lactation number. Based on the odds ratio, treated cows were 2.4 (1/0.41) times less likely to miscarry, whereas the risk of pregnancy loss was 1.6 times higher in cows that became pregnant during the warm period in comparison to the cool period. These results support the hypothesis that sub-optimal progesterone concentrations in high producer dairy cows may compromise conceptus development. Under these conditions, intra-vaginal progesterone supplementation has the potential to reduce the incidence of pregnancy loss during the early fetal period.     

Radioimmunoassay of a bovine pregnancy-associated glycoprotein in serum: its application for pregnancy diagnosis.

A sensitive and specific double-antibody RIA for a bovine pregnancy-associated glycoprotein (bPAG) is described. The limit of detection was 0.2 ng/ml. The assay was specific for bPAG in that pituitary and placental gonadotropic hormones and other placental or serum proteins assayed in serial dilutions did not cross-react. The RIA allowed measurement of bPAG in placental extracts, fetal serum, fetal fluids, and serum or plasma of pregnant cows. About 20% of unbred heifers and nonpregnant cows had detectable levels ranging from 0.30 +/- 0.09 to 0.50 +/- 0.17 ng/ml (mean +/- SD), and 15% of bull sera showed higher concentrations (3.01 +/- 1.73 ng/ml) of bPAG or bPAG-like protein. Variations among animals was observed in fetal serum bPAG concentrations. Bovine PAG was detected in maternal peripheral blood at Day 22 of pregnancy (mean +/- SD, 0.38 +/- 0.13 ng/ml) in some animals and at Day 30 in all pregnant cows. Peripheral serum bPAG levels increased progressively to 3.60 +/- 1.73 ng/ml (mean +/- SD) at Day 30 of pregnancy, to 24.53 +/- 8.81 ng/ml at Day 120, and to 1551.91 +/- 589.68 ng/ml at Day 270. Peak concentration of bPAG was 2462.42 +/- 1017.88 ng/ml and it occurred 1-5 days prior to parturition. After delivery, bPAG concentrations decreased steadily to 499.63 +/- 267.20 ng/ml at Day 14 postpartum (pp), 10.12 +/- 7.84 ng/ml at Day 60 pp, and 1.44 +/- 1.08 ng/ml at Day 90 pp. The undetectable concentration (less than 0.20 ng/ml) was reached by Day 100 +/- 20 pp. An investigation undertaken in Holstein heifers, Holstein cows, and Hereford cows used as recipients for purebred Holstein embryos supplied evidence of the influence of breed of recipient and sex of fetuses on peripheral concentrations of bPAG. A herd of 430 Holstein-Friesian heifers that had received transferred embryos were bled at Day 35 postestrus (pe) for measurement of bPAG. The bPAG was detected in 287 of 430 serum samples analyzed. By rectal palpation performed at Day 45 pe, 267 heifers with detectable levels of bPAG at Day 35 pe were confirmed to be pregnant as were 3 of 143 heifers previously diagnosed as not pregnant by RIA. These results suggest that detection of this placental-specific antigen in the serum could be used as a specific serological method for early pregnancy diagnosis in cattle from 28 days after breeding.     

Pregnancy rates relative to recipient plasma progesterone levels on the day of nonsurgical transfer of frozen/thawed bovine embryos

A total of 71 synchronized dairy heifers (Holstein Friesian x German Black Pied) were used as recipients of seven-day old frozen/thawed bovine embryos. Plasma progesterone concentrations and corpus luteum quality on the day of nonsurgical transfer (= day 7) were determined and related to pregnancy rates or estrus intervals in nonpregnant recipients. A total of 32 recipients (45.1 %) maintained pregnancy; 39 recipients (54.9 %) did not. No significant differences could be detected between progesterone levels in recipients that remained pregnant (3.14 +/- 0.24 ng/ml; x +/- SEM ) and those that did not maintain pregnancy (3.23 +/- 0.28 ng/ml). Optimal progesterone levels were between 2 and 5 ng/ml coinciding with a pregnancy rate of 51.1 % (24 47 ). Pregnancy rates apparently were decreased when progesterone levels were below 2 ng/ml (35.3 %; 6 17 ) or above 5 ng/ml (28.6 %; 2 7 ). Hence, optimal progesterone levels were identical to those for freshly collected embryos reported previously by Remsen et al. (1). Bovine corpus luteum quality graded by rectal palpation was related to some extent to progesterone levels but not to pregnancy rates. Out of 39 nonpregnant recipients seven animals (17.9 %) with a mean plasma progesterone level of 3.76 +/- 0.72 ng/ml showed an extended estrus interval of more than 55 days, probably indicating early embryonic mortality. Progesterone levels did not significantly differ between nonpregnant recipients with estrus intervals of various length. Plasma progesterone levels at the time of transfer are of limited diagnostic value for screening recipients prior to transfer of frozen/thawed embryos.     

A simple enzymeimmunoassay of milk progesterone for monitoring fertility in dairy buffaloes

A simple, sensitive, direct (without extraction) enzymeimmunoassay (EIA) was usec to determine progesterone levels in whole milk samples of 400 Nili-Ravi dairy buffaloes. The mean progesterone values 22 d after A.I. were significantly higher in pregnant (16.6 +/- 9.2 ng/ml) than nonpregnant (below 5 ng/ml) animals. The mean progesterone values were below 0.34 +/- 0.12 (the detection limit) both at estrus and in cases of clinically diagnosed inactive ovaries, 3.18 +/- 1.9 at proestrus, 2.25 +/- 1.2 postestrus and 13.22 +/- 6.74 at Day 10 of the estrous cycle. Twenty buffaloes confirmed pregnant for 2 to 3 mo, had a mean value of 20.3 +/- 4.5 ng/ml. The EIA test is very reliable in the selection of nonpregnant buffaloes (100 %) and the confirmation of inactive ovaries and of estrus. Differential diagnosis of inactive or active ovaries can be made by analyzing two milk samples at a 7-d interval.     

Pregnancy-specific protein B and progesterone concentrations in French Alpine goats throughout gestation.

The 34 French Alpine dairy goats originated from a single flock and were artificially inseminated 44 h after synchronization of oestrus. They were bled daily at the jugular vein from 15 to 27 days after AI. An early pregnancy diagnosis by RIA of progesterone concentration was performed 21 days after AI. In pregnant goats (greater than 1.5 ng progesterone/ml) daily sampling was extended until 30 days after AI and, from those, 9 were bled every 2 weeks until the end of pregnancy and at 50 and 63 days post partum. Pregnancy-specific protein B (PSPB) was also assayed. The kidding rate was 67.6% (23/34). PSPB concentrations (ng/ml) in pregnant goats were significantly different from those of non-pregnant goats at 24 days after AI (0.82 +/- 0.18 vs 1.78 +/- 0.19; mean +/- s.e.m.) and rose to 40 ng/ml at the end of pregnancy. From Day 25 and throughout gestation, females with 2 fetuses had higher PSPB concentrations than did those with a single fetus (P less than 0.05). In the 2 goats exhibiting late embryonic mortality according to progesterone concentrations, one had a PSPB profile very similar to those of pregnant goats until 30 days while the other did not show any elevation of PSPB concentration. It is concluded that PSPB profiles in goats are similar to those found in cows throughout pregnancy and that PSPB RIA may be useful for pregnancy diagnosis or diagnosis of late embryonic mortality.     

The accuracy of enzyme-linked immunosorbent assay and latex agglutination progesterone test for the validation of estrus and early pregnancy diagnosis in dairy cattle

The accuracy of the enzyme-linked immunosorbent assay (ELISA) and the latex agglutination (LA) on-farm progesterone kit for detecting estrus and diagnosing early pregnancy was investigated in this study. Italian Friesian dairy cows (n=82) from 6 dairy herds were used for the collection of foremilk samples at the time of breeding and at 19, 21, and 23 days post insemination. Pregnancy status was ascertained by uterine palpation per rectum 40 to 60 days post insemination. Progesterone levels were affected by herd, percentage of milk fat, and the day of testing x diagnosis interaction. Validation of estrus by qualitative on-farm tests was 74.6% (LA) and 100.0% (ELISA) accurate using 0.5 ng/ml of progesterone as the RIA estimate for estrus. The accuracy rate for early pregnancy diagnosis by RIA was 68.4 to 83.8% for day 19 and day 21, respectively, while the detection rate for nonpregnancy was 84.6 to 100% on day 19 and day 21, respectively, as compared with uterine palpation per rectum. The average accuracy rate for early pregnancy diagnosis ranged from 84.7 to 92.3% for the LA and ELISA tests, respectively; the nonpregnancy rate was correctly predicted 93.9% to 68.2% for the LA and ELISA tests, respectively.     

Serum progesterone concentration in pyometritic and normal postpartum dairy cows

Two studies were conducted to evaluate the concentration of serum progesterone in pyometritic cows and relate it to palpation of ovarian structures per rectum . In Trial 1, serum samples from 34 pyometritic cows were assayed for progesterone. Mean serum progesterone concentrations were 6.8 +/- 0.7 ng/ml. In Trial 2, each of 54 pyometritic cows was paired with a control cow on the basis of days post partum (18-50 days). Mean concentration of progesterone was 9.7 +/- 1.0 ng/ml for the pyometritic cows and 5.7 +/- 0.8 ng/ml in control cows (P<0.005). Progesterone concentration was greater (P<0.005) in both groups of cows with palpable corpora lutea (CLs). Ninety-six percent of the pyometritic cows had palpable CLs compared to 57% of the control group. Comparing serum progesterone only in cows with a palpable CL, the mean concentration was still greater (P<0.005) in the pyometritic group (10.6 +/- 1.0 ng/ml) than the control group (6.6 +/- 1.0 ng/ml). Compatability of rectal palpation findings and concentrations of serum progesterone were 92.6% for the pyometritic group and 72.2% for the control group. Progesterone concentration increased (P<0.05) by increased days post partum in Trial 2 (n=54) but not in Trial 1 (n=23). In both Trials 1 and 2, uterine size due to pyometra increased (P<0.05) with increased days post partum. No other associations were found.     

Diagnosis of pregnancy by radioimmunoassay of a pregnancy-specific protein in the plasma of dairy cows

The accuracy and efficiency of progesterone (P4) and bovine pregnancy-specific protein B (bPSPB) radioimmunoassays (RIA) in detecting pregnant and nonpregnant dairy cows were compared at different stages of pregnancy. The study included 145 French Friesian heifers and cows from a single herd. A total of 175 artificial insemination (A.I.) and blood sampling procedures were performed. Animals were bled 24 d post AI for P4 RIA. They were bled at 24, 26, 30 to 35, and 70 +/- 9 after AI for bPSPB RIA. Females were declared nonpregnant when plasma P4 concentrations were lower than 1.5 ng/ml. With the bPSPB RIA, cows were nonpregnant when at least one of the B Bo x 100 replicates was higher than 95% in the RIA. When compared with palpations per rectum at 70 d, the accuracy of positive diagnoses (no. positive and pregnant/no. positive diagnoses) by P4 RIA at Day 24 was 67.2% (82 122 ). The accuracy of negative diagnoses was 98% (52 53 ). Accuracy of positive diagnoses by bPSPB RIA increased with gestation age (P<0.05) from 86.2% (50 58 ) on Day 24 to 98.8% (83 84 ) at time of palpation per rectum. Accuracy of negative diagnoses increased (P< 0.001) from Day 24 (71.8%; 84 117 ) to Days 30 to 35 (100%, 83 83 ). Efficiency in detecting nonpregnant females was much higher (P < 0.001) with the bPSPB RIA on Days 30 to 35 (90.2%; 83 92 ) than with the P4 RIA on Day 24 (56.5%, 52 92 ). It is concluded that 30 days after AI, the bPSPB RIA is an efficient test both for pregnancy prediction and detection of nonpregnant dairy cows.     

Genetic and environmental factors influencing first service conception rate and late embryonic/foetal mortality in low fertility dairy herds

The objective of this study was to identify factors affecting variation in conception rate to first artificial inseminations (AI) (CR: number of pregnant cows on D80-100/inseminated cows) and the incidence of embryonic/foetal loss (LEM) between 21 and 80 days of pregnancy (number of cows non-pregnant on D80-100/pregnant on D21) in 44 low fertility dairy herds of the west-central region of France. Reproductive status was assessed using progesterone milk concentration on D0 = Day of AI and D21-24, plasma PSPB concentration on D30-35, rectal palpation on D80-100 and observed return to oestrous. The final data set contained 1285 Prim'Holstein cows, 5.0% (64/1285) were inseminated in the luteal phase (progesterone > or = 3 ng/ml on D0), 61.3% (787/1285) were pregnant on D21-24 (progesterone < 3 ng/ml on D0 and > or = 5 ng/ml on D21-24), 15.4% lost their embryo/foetus between D21-24 and D80-100 (198/1285) and 45.8% (589/1285) were pregnant on D80-100. The incidence of late embryonic/foetal loss (LEM) was 25.2% (198/787). Multivariate logistic regression models including the random herd effect were used to analyse the relationship between AI centre, AI sire, cow's sire, parity, interval between calving and AI, milk production, milk protein content, body condition score (BCS) on D0, season of calving, season of AI, estimated genetic index on CR and LEM incidence. CR was significantly related to parity (p < 0.05), milk production after calving (p < 0.05) and estimated genetic value (p < 0.01). A significant difference in CR was observed for calving to AI interval > or = 70 days versus > or = 90 days, but the overall effect of the interval was not significant (p = 0.11). LEM incidence was affected by period of AI (p < 0.05), milk production (p < 0.05) and BCS (p < 0.05), but was not related to estimated genetic index. In conclusion, in these low fertility herds, the incidence of LEM was high and 25% of the cows lost their embryo after 21 days of pregnancy. LEM was affected by specific factors (season, BCS), which were not related to CR. The absence of a relationship between estimated genetic index and LEM in spite of its effect on CR indicates that estimated genetic merit has a greater effect on early embryonic loss or fertilisation failure than on later stages of embryo development.     

Conception rates and serum progesterone concentration in dairy cattle administered gonadotropin releasing hormone 5 days after artificial insemination

The objectives of this study were to determine the effect of administration of exogenous GnRH 5days after artificial insemination (AI) on ovarian structures, serum progesterone concentration, and conception rates in lactating dairy cows. In experiment 1, 23 Holstein cows were synchronized using the Ovsynch protocol. Five days after AI (day 0) cows were assigned randomly to receive either saline (saline; n=11) or 100microg GnRH (GnRH; n=12). To examine ovarian structures, ultrasonography was performed on day 1 and every other day beginning on day 5 until day 13. On days 5 and 13 blood samples were obtained to measure serum progesterone concentrations. All cows in the GnRH-treated group developed an accessory corpus luteum (CL), whereas cows in the saline group did not. Mean serum progesterone concentrations did not differ between GnRH and saline groups on day 5 (1.64+/-0.46ng/ml versus 2.04+/-0.48ng/ml). On day 13 serum progesterone concentrations were greater (P<0.05) in the GnRH group compared with saline (5.22+/-0.46ng/ml versus 3.36+/-0.48ng/ml). In experiment 2, 542 lactating cows, at two different commercial dairies, were used to test the effect of administering GnRH 5 days after AI on conception rates. Cows were synchronized and detected for estrus according to tail chalk removal. Cows detected in estrus received AI within 1h after detection of estrus. Five days after AI, cows were assigned randomly to receive either GnRH (n=266) or saline (n=276). Pregnancy status was determined by palpation per rectum of uterine contents approximately 40 days after AI. There was no effect of farm on conception rate. There was no effect of treatment as conception rates did not differ between GnRH and saline groups (26.7% GnRH versus 24.3% saline). Regardless of treatment, days in milk, parity, milk yield, and number of services had no effect on the odds ratio of pregnancy. In summary, the results of this study indicated that GnRH administered 5 days after AI increased serum progesterone by developing an accessory CL but did not improve conception rates in dairy cattle.     

Increase in milk and body temperature of cows as a sign of embryo entry into the uterus

The objective of the first stage of these studies was to investigate whether temperature increases in the milk and body of cows during the early period of pregnancy. We studied 94 pregnant and 116 nonpregnant cows, and the temperatures were measured daily beginning 24 days after insemination. In addition, progesterone levels in milk were measured twice (on Days 21 and 24 after insemination), and examinations per rectum were conducted to determine pregnancy. Data analysis showed that in almost 90% of the pregnant cows milk temperature increased by 0.64 degrees C some time between Days 5 and 12 after insemination. This increase in milk temperature was highly significant and corresponded with a 0.46 degrees C increase in body temperature. There was no increase in the milk or body temperature in nonpregnant cows. After discovering in the first stage that a high percentage of pregnant cows experienced both milk and body temperature increases, we focused our attention in the second stage of the study on the question of whether this increase in temperatures might be due to an immune response of the mother to the entry of the embryo into the uterus. In this stage we conducted three different experiments on another group of animals consisting of 309 cows and heifers. The experiments included analysis of progesterone, estradiol, cortisol and PGE2 levels in the blood serum of the cows and heifers; an estimation of the pyrogenic activity of PGE1, PGE2 and progesterone; and measurements of body temperature in the heifers before and after embryo transfer. The results of these experiments suggested that an increase in milk and body temperature could be an indicator of an immune response of the mother to the entry of the embryo into the uterus.     

Milk progesterone measurement in dairy cows: Correlation with estrus and pregnancy determination

Progesterone levels in fore milk, determined by a highly specific radioimmunoassay, were compared for the assessment of estrus by a veteran herdsman and an experienced inseminator, in cows presented for insemination. In addition, an examination was made of the relative accuracy of using milk progesterone levels for the determination of pregnancy at 24, 40 and 44 days after insemination, as compared with rectal palpation at 45–50 days post-breeding.Fat-free fore milk progesterone levels were similar to jugular plasma levels at 24 days post-insemination and reached roughly 60% of the level of unextracted fore milk at this time. Accuracy of estrus diagnosis by herdsman, inseminator and milk progesterone level was 84%, 93% and 96%, respectively. For pregnancy diagnosis, milk progesterone determination in 85 cows showed 78% accuracy in predicting pregnancy and 100% accuracy in predicting non-pregnancy. At 40 days post-insemination false positives dropped to 10% and at 44 days only 7% of the cows were incorrectly diagnosed as pregnant. The false positives in this study were largely due to embryonic mortality as reflected by abnormal intervals of return to estrus. Two milk progesterone determinations, at 24 and either 40 or 44 days post-insemination ensure maximum reliability for early pregnancy diagnosis.     

The predictive value, sensitivity and specificity of palpation per rectum and transrectal ultrasonography for the determination of bovine luteal status

Luteal status (classifications 1 and 3 = high progesterone levels, 2 and 4 = low progesterone levels) was diagnosed in randomly cyclic Holstein cows, using palpation per rectum and B-mode transrectal ultrasonography, respectively. The results were compared in 2x2 contingency tables with radioimmunoassay (RIA) classifications. Cows with milk progesterone levels of 5.0 ng/ml or higher were classified high and cows with levels below 5.0 ng/ml of progesterone were low. The predictive value, sensitivity and specificity results were 1) 88.2, 68.2 and 88.9% for Classification 1; 69.6, 88.9 and 68.2 for Classification 2; 90.9, 83.3 and 87.5 for Classification 3; and 77.8, 87.5 and 83.3 for Classification 4. Within each test, the diagnosis of high progesterone status was more accurate than of low status. The types of errors are discussed. A correlation coefficient of 0.68 resulted between RIA progesterone concentration and ultrasonographic luteal diameter. Ultrasonography was a better diagnostic test than palpation per rectum.     

Effectiveness of prostaglandin F2alpha in the initial treatment of bovine ovarian cysts

This study was conducted to evaluate the use of prostaglandin F2alpha (PGF2alpha) in the initial treatment of ovarian cysts in dairy cattle. Two hundred and sixty three cows diagnosed cystic on palpation per rectum were randomly assigned to one of three treatment groups (A, B or C). Cows in Groups A and B were treated with 25 mg i.m.of PGF2alpha at the time of diagnosis (Day 0), while cows in Group C received 100 mug of GnRH. Seven days following initial treatment (Day 7), cows from Group A that were not observed in estrus were treated with GnRH. Cows from Groups B and C were not treated. On Day 14, all cows that had not been inseminated received PGF2alpha. A blood sample was obtained from all cows on Days 0, 7 and 14 and was analyzed for progesterone (P4) using radioimmunoassay. Incidences of estrus were recorded and cows that were more than 60 d in milk at the time of diagnosis were bred when observed in estrus. The incidence of follicular cysts on Day 0 (as defined as P4 <0.5 ng/ml) was similar between groups and constituted about 40% of all cysts. There were significantly more cows pregnant to insemination within 7 d of initial treatment in Group B than in Groups A and C (P<0.05). After Day 14, the pregnancy rate was not statistically different between Group B and C, but Groups B and C had a statistically higher pregnancy rate than Group A from Day 21 to Day 35. At the end of the study, there was no statistical difference for the pregnancy rate between groups. We concluded that treatment of ovarian cysts diagnosed by per rectum examination with prostaglandin (at time of diagnosis and 14 d later for cows that were not inseminated) was as effective as initial treatment with GnRH followed by prostaglandins 14 d later for cows that were not inseminated previously. Cows that were initially treated with prostaglandins also tended to become pregnant sooner.     

Milk progesterone levels to monitor reproductive status of Murrah buffalo

Concentration of progesterone in whole milk was used to diagnose pregnancy in 44 lactating buffaloes. Milk samples were taken from days 19 to 27 post breeding and analysed for progesterone by radio-immunoassay. A level exceeding 10 ng/ml of milk was taken as an indication of pregnancy. Using this criterion, the accuracy of the pregnancy test from a single milk sample on 19, 21, 23, 25 and 27 days after insemination ranged from 71.42 to 86.95% and 86 to 87.50% for pregnant and non-pregnant animals, respectively. Milk progesterone concentrations were significantly higher (P / 0.001) in pregnant compared with those in non-pregnant buffaloes on day 19, 21, 23, 25 and 27 post-insemination.     

Effects of an analog of PGF2(alpha) (ONO-1052) on cows with luteinized ovarian cysts following treatment with GnRH analog (fertirelin acetate)

Effects of PGF2(alpha) analog (ONO-1052) on cows with luteinized cysts following treatment with GnRH analog (fertirelin) in inducing estrus and shortening an interval from treatment to conception were investigated in a field trial. Seventy-five cows with follicular cysts diagnosed by rectal palpation were treated intramuscularly (i.m.) with 400 mug fertirelin (an analog of GnRH). Luteinization of follicular cysts were tentatively judged by rectal palpation. Forty-one cows were considered to have luteinized cysts and were either treated i.m. with ONO-1052 (28 cows) or left untreated as controls (13 cows). Thirty-four other cows were considered to have not responded to fertirelin and retreated with 10,000 MU hCG (19 cows) or fertirelin (five cows). This tentative judgment of luteinization of the cysts by rectal palpation after treatment was later confirmed by determining serum progesterone concentration before and 10 to 14 days after treatment. Only in the cows with luteinized cysts which were confirmed by serum progesterone analysis; increased from a pretreatment level below 1.0 ng/ml to a value of 1.0 ng/ml or higher 10 to 14 days after treatment, effects of ONO-1052 combined with fertirelin were investigated. Of the 18 cows with luteinized cysts thus confirmed following fertirelin injection and treated with ONO-1052, 15 (83.3 %) cows came into estrus within 26 +/- 14 (mean +/- SD) days after the first treatment; eight cows within three to five days and four cows within 22 to 28 days, and 14 cows (77.8 %) conceived within 100 days (42 +/- 26 days). On the other hand, the five control cows with luteinized cysts that were not treated with ONO-1052 required significantly longer to exhibit normal estrus (54 +/- 13 days; P<0.05) and to conceive (54 +/- 13 days). These results indicate that ONO-1052 combined with fertirelin may be useful to shorten the interval from treatment to normal estrus and to conception in cows with follicular cysts.