Factors affecting plasma progesterone in the early fetal period in high producing dairy cows

The aim of the present study was to determine whether certain animal- or management related factors could affect plasma progesterone concentrations on Day 42 of gestation in high producing dairy cows. Factors affecting early fetal loss were also analyzed. The study population was comprised of 199 pregnant cows classified as having high (> or =9 ng/ml) or low (<9 ng/ml) plasma progesterone concentrations. Through logistic regression procedures it was determined that, based on the odds ratio, cows with two or more corpora lutea were three times more likely to have high progesterone concentrations than cows with a single corpus luteum. Low producing cows during the warm season were 2.86 times more likely to have high progesterone concentrations than the remaining cows. Primiparous cows with high concentrations (>4 ng/ml) of pregnancy associated glycoprotein-1 (PAG-1) were 2.73 times more likely to have high progesterone concentrations than the remaining cows. Of the 199 pregnancies, 25 (12.6%) suffered early fetal loss: 22/136 (16.2%) during the warm season and 3/63 (4.8%) during the cool season, all in cows without additional corpora lutea. Based upon the odds ratio, cows without an additional corpus luteum were 3.67 times more likely to suffer fetal loss during the warm season than during the cool season. Our results indicate that milk production, the presence of two or more corpora lutea and plasma PAG-1 concentrations can affect plasma progesterone concentrations at the onset of the fetal period. The presence of an additional corpus luteum strongly diminished the risk of early fetal loss during the warm period.     

Relationship between heat stress during the peri-implantation period and early fetal loss in dairy cattle

The aim of the present study was to establish whether temperature-humidity index values, as a measure of heat comfort, from Days 1 to 40 of gestation could be associated with the pregnancy loss rate in high producing dairy cows. Data from 1391 pregnancies were recorded. Pregnancy was diagnosed by transrectal ultrasonography between Days 34 and 45, and again 90 days after insemination. Pregnancy loss was assumed when the second pregnancy diagnosis on Day 90 proved negative and was registered in 7.8% (108/1391) of pregnancies. Mean and maximum temperature-humidity index values were established for each cow for Days 0 (day of insemination), 1, 2 and 3 after insemination, and averages established for Days 0-3, 0-10, 11-20, 21-30 and 31-40 after insemination. Cow and management variables previously found to be significantly correlated with the early fetal loss in the same geographical area were also recorded. The relative contribution of each factor to the probability of pregnancy loss was determined using logistic regression models. Based on the odds ratio, a strong association with pregnancy loss of the factors warm period of pregnancy (warm period-May to September versus cool-October to April), twin pregnancy (as negative factors: odds ratios 3.1 and 3.4, respectively) and an additional corpus luteum (as a positive factor: odds ratio 0.32) was confirmed. The likelihood of pregnancy loss increased by a factor of 1.05 for each additional unit of the mean maximum temperature-humidity index from Days 21 to 30 of gestation. Logistic regression analysis revealed no significant effects of temperature-humidity index values for the remaining gestation periods. Our results indicate that heat stress can compromise the success of gestation during the peri-implantation period, such that high temperature-humidity index values for the period 21-30 days of gestation are a risk factor for subsequent early fetal loss.     

Clinical implications of induced twin reduction in dairy cattle

Embryo reduction may prevent the negative effects of twinning in dairy cattle; however, the technique may carry an additional risk of pregnancy loss. The aim of this study was to evaluate the effect on pregnancy maintenance of embryo reduction by manual amnion rupture in unilateral and bilateral twin pregnant cows. A secondary objective was to examine the dynamics of endocrine factors following the treatment. On Day 35-41 of gestation 55 cows bearing two live twin embryos (28 bilateral, 27 unilateral) were randomly assigned to a twin reduction group (n = 27; cows fitted with a progesterone releasing intra-vaginal device for 21 days after manual amnion rupture) or control group (n = 28; untreated cows). Pregnancy loss before Day 90 was recorded in nine control and eleven twin reduction cows (32.1% vs 40.7%, respectively, p = 0.508). Logistic regression models indicated that laterality was the only variable significantly affecting pregnancy loss. The pregnancy loss risk was 8.7 times higher for unilateral than for bilateral twin pregnancies (59.3% vs 14.3%, respectively, P < 0.001) yet was similar in the unilateral control and unilateral twin reduction cows (62.3% vs 53.8%, respectively, P = 0.581). In contrast, four of 14 cows with bilateral twin pregnancies undergoing twin reduction lost their pregnancies while no losses were recorded in control cows with bilateral pregnancies (P = 0.049). A rise in plasma progesterone concentration was detected on the day following treatment in the twin reduction group and concentrations remained high within the first week of treatment. Plasma pregnancy-associated glycoprotein-1 (PAG-1) concentrations fell between Day 35-41 and Day 42-48, regardless of treatment. Our findings indicate that embryo reduction by manual amnion rupture did not carry an additional risk of pregnancy loss for unilateral twin pregnancies, whereas it increased the risk of pregnancy failure in bilateral twin pregnancies. However, benefits of preventing cows from delivering twins might also be considered when assessing the success of embryo reduction in bilateral twin pregnancies.     

Plasma pregnancy-associated glycoprotein-1 (PAG-1) concentrations during gestation in Neospora-infected dairy cows

The aim of this study was to determine whether plasma pregnancy-associated glycoprotein-1 (PAG-1) concentrations in pregnancy are affected by persistent Neospora caninum infection in dairy cows. The data analyzed were derived from 22 multiparous cows: 16 N. caninum-seropositive and 6 N. caninum-seronegative animals (used as controls). Three of the 16 seropositive cows aborted during the study period and the corresponding data were analyzed separately. Pregnancy diagnoses were performed on day 40 post-insemination by transrectal ultrasound, and by palpation per rectum on days 90, 120, 150, 180 and 210. Blood samples were collected from each animal immediately before each pregnancy diagnosis, and then at parturition or at the time of abortion detection. Plasma was tested for antibodies against N. caninum and PAG-1 concentrations were determined by radioimmunoassay. In non-aborting animals, the effects of neosporosis (seropositive versus seronegative), N. caninum antibody levels, semen providing bull, sex of the newborn, and day of gestation on PAG-1 concentrations were evaluated by GLM repeated measures analysis of variance. The effect of the gestation period (first half versus second half) on the N. caninum antibody titer was established by the Student's t-test in seropositive cows. A significant positive effect of gestation day on PAG-1 concentrations was observed (d.f.=6; F=12.6; P<0.0001). For all cows, PAG-1 concentrations increased steadily during the course of gestation, with peak concentrations recorded at parturition. Neosporosis (P=0.493), N. caninum antibody levels (P=0.921), sex of the newborn (P=0.856) and semen providing bull (P=0.087) had no effect on plasma PAG-1 concentration. There was a significant 52% increase (P<0.0001) in N. caninum antibody titers during the second half of gestation compared to the first half. The fates of the three aborting cows were abortion on gestation day 215 in one, and fetus mummification diagnosed on gestation days 180 and 210, respectively, in the remaining two cows. A luteolytic dose of prostaglandin was applied 30 days after mummification diagnosis in these last two cows, and fetus expulsion was detected on days 215 and 250, respectively. Two of the aborted fetuses were submitted to laboratory analysis and the presence of N. caninum was confirmed by specific PCR. In the cows with a mummified fetus, PAG-1 concentrations were low or undetectable when the diagnosis was made. These findings suggest that N. caninum infection has no effect on placental function in chronically infected, cows not suffering abortion, while PAG-1 measurements in aborting animals provide a useful indication of feto-placental status.     

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.     

Spontaneous reduction of advanced twin embryos: its occurrence and clinical relevance in dairy cattle

Twin pregnancies represent a management problem in dairy cattle since the risk of pregnancy loss increases, and the profitability of the herd diminishes drastically as the frequency of twin births increases. The aim of this study was to monitor the development of 211 twin pregnancies in high producing dairy cows in order to determine the best time for an embryo reduction approach. Pregnancy was diagnosed by transrectal ultrasonography between 36 and 42 days after insemination. Animals were then subjected to weekly ultrasound examination until Day 90 of gestation or until pregnancy loss. Viability was determined by monitoring the embryonic/fetal heartbeat until Day 50 of pregnancy, and then by heartbeat or fetal movement detection. Eighty-six cows (40.8%) bore bilateral and 125 (59.2%) unilateral twin pregnancies. Embryo death was registered in one of the two embryos in 35 cows (16.6%), 33 of them at pregnancy diagnosis. Pregnancy loss occurred in 22 of these cows between 1 and 4 weeks later. Thus, 13 (6.2% of the total animals) cows, carrying one dead of the two embryos, maintained gestation. Total pregnancy loss before Day 90 of pregnancy (mean 69 +/- 14 days) was registered in 51 (24.2%) cows: 7 (8%) of bilateral pregnancies and 44 (35.2%) of unilateral pregnancies, and it was higher (P = 0.0001) for both right (32.4%, 24/74) and left (39.2%, 20/51) unilateral than for bilateral (8.1%, 7/86) twin pregnancies. The single embryo death rate was significantly (P = 0.02) lower for cows with bilateral twins (9.3%, 8/86) than for total cows with unilateral twins (21.6%, 27/125). By way of overall conclusion, embryo reduction can occur in dairy cattle, and the practical perspective remains that most embryonic mortality in twins (one of the two embryos) occurs around Days 35-40 of gestation, the period when pregnancy diagnosis is generally performed and when embryo reduction could be tried.     

Factors affecting pregnancy loss for single and twin pregnancies in a high-producing dairy herd

Our objective was to determine the magnitude of, and factors affecting, pregnancy loss for lactating Holstein cows on a commercial dairy farm when diagnosed with twin (n = 98) or single (n = 518) pregnancies using transrectal ultrasonography. Pregnancy losses were assessed with records of non-viable embryos at first pregnancy examination and embryo losses between the first (25-40 d after AI) and second (48 and 82 d after AI) post-breeding pregnancy examinations. Among cows diagnosed with single pregnancies, 3.7% were diagnosed with a non-viable embryo at first pregnancy examination, and 4.6% of those diagnosed with a viable embryo underwent pregnancy loss by the second examination. A total of 11.2% of cows diagnosed with twins experienced a single embryo reduction, whereas 13.3% lost both embryos. Overall, the total proportion of cows experiencing pregnancy loss or experiencing embryo reduction was greater for cows diagnosed with twin than single pregnancies (odds ratio; OR = 3.6), resulting in an embryo survival rate of 91.9% for cows diagnosed with single compared to 75.5% for cows diagnosed with twin pregnancies. Season of breeding and milk production were associated with pregnancy loss for single pregnancies, whereas CL number was associated negatively with embryo reduction and pregnancy loss for twin pregnancies. The risk of twinning and double ovulation among pregnant cows increased with days in milk (DIM), and the risk of double ovulation was greater for cows diagnosed with ovarian cysts and lacking a CL at initiation of an Ovsynch protocol. We concluded that in this herd, embryo reduction and pregnancy loss during early gestation was greater for lactating Holstein cows diagnosed with twin compared to single pregnancies. In addition, cows diagnosed with ovarian cysts and lacking a CL had an increased risk for double ovulation.     

Effects of cumulative stressful and acute variation episodes of farm climate conditions on late embryo/early fetal loss in high producing dairy cows

The aim of this study was to determine possible relationships between farm climate conditions, recorded from day 0 to day 40 post-artificial insemination (AI), and late embryo/early fetal loss in high producing dairy cows. Pregnancy was diagnosed by rectal ultrasonography between 28 and 34 days post-AI. Fetal loss was registered when a further 80- to 86-day diagnosis proved negative. Climate variables such as air temperature and relative humidity (RH) were monitored in the cubicles area for each 30-min period. Temperature-humidity indices (THI); cumulative stressful values and episodes of acute change (defined as the mean daily value 1.2 times higher or lower than the mean daily values of the 10 previous days) of the climate variables were calculated. The data were derived from 759 cows in one herd. A total of 692 pregnancies (91.2%) carried singletons and 67 (8.8%) carried twins. No triplets were recorded. Pregnancy loss was recorded in 6.7% (51/759) of pregnancies: 5.6% (39/692) in single and 17.9% (12/67) in twin pregnancies. Using logistic regression procedures, a one-unit increase in the daily cumulative number of hours for the THI values higher than 85 during days 11–20 of gestation caused a 1.57-fold increase in the pregnancy loss, whereas the likelihood of fetal loss increased by a factor of 1.16 for each additional episode of acute variation for the maximum THI values during gestation days 0–40. THI values higher than 85 and episodes of acute variation for the maximum THI values were only recorded during the warm and cool periods, respectively. The presence of twins led to a 3.98-fold increase in pregnancy loss. In conclusion, our findings show that cumulative stressful and episodes of acute variation of climatic conditions can compromise the success of gestation during both the cool and warm periods of the year. Twin pregnancy was confirmed as a main factor associated with pregnancy loss.     

Relationship of pre-ovulatory follicle size, estradiol concentrations and season to pregnancy outcome in dairy cows

This study was carried out to evaluate effects of pre-ovulatory follicle size, plasma concentrations of estradiol and progesterone, and season on pregnancy outcomes in dairy cows. Holstein cows (n = 144) were synchronized and inseminated (Ovsynch/TAI protocol) in two distinct periods (cold versus warm season). Blood samples were collected daily from AI (day 0) to day 8 and on days 15, 22, 29, 36 and 64 to measure progesterone and estradiol. Pregnancy diagnosis was performed at days 29, 43 and 64. The pre-ovulatory follicle size was larger and the plasma estradiol concentrations on the day of AI were greater in animals that became pregnant. Plasma progesterone concentrations diverged and became greater after day 5 post-AI, in cows diagnosed pregnant, as compared to non-pregnant cows. The overall pregnancy rate (33%) or late embryonic/early fetal losses (23%) did not differ between seasons, but plasma estradiol concentrations on the day of AI and plasma concentrations of progesterone in pregnant cows were lower in the warm season. Reduced CL function, measured as plasma progesterone concentrations, from days 22 or 29 post-AI onward for cold and warm season, respectively, was associated with subsequent late embryonic/early fetal mortality. Overall, pregnancy was related to diameter of the pre-ovulatory follicle and plasma E2 on the day of AI, but embryonic/fetal losses were not. Season did not affect these outcomes, even though it influenced luteal function after AI.     

Risk of minor and major fetal malformations in diabetics with high haemoglobin A1c values in early pregnancy.

Maternal haemoglobin A1c (HbA1c) values were measured before the end of the 15th week of gestation in 142 pregnancies in women with insulin dependent diabetes. In pregnancies complicated by fetal malformations (n = 17) the mean initial HbA1c value was 9.5 (SD 1.8)% of the total haemoglobin concentration, which was significantly (p less than 0.001) higher than in pregnancies without malformations (8.0 (SD 1.4)%; n = 125). HbA1c values did not differ between pregnancies complicated by minor and major fetal malformations, but the rate of malformations showed a positive relation to the HbA1c value in early pregnancy (chi 2 = 11.9; p = 0.001). Fetal malformations occurred in six out of 17 pregnancies (35.3%) in mothers whose initial HbA1c value was 10% or more, in eight out of 62 pregnancies (12.9%) in mothers with initial values between 8.0% and 9.9%, and in only three out of 63 pregnancies (4.8%) in mothers with an initial value below 8.0%. These data support the hypothesis that the increased incidence of fetal malformations in mothers with insulin dependent diabetes is associated with maternal hyperglycaemia during organogenesis. Hence diabetic women who are planning to have a child--especially those with a high HbA1c value--should receive intensified metabolic control.     

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).     

Neospora caninum infection does not affect early pregnancy in dairy cattle

This study was designed to evaluate the relationship between Neospora caninum infection prior to pregnancy, as determined through maternal serology, and the subsequent occurrence of abortion in dairy cattle. Special emphasis was placed on pregnancy losses in the first trimester of pregnancy. Neospora caninum antibodies were analyzed by commercial ELISA in 2773 pregnant animals (2022 parous cows and 751 heifers) from six herds. The mean seroprevalence of antibodies to N. caninum in the herds was 15.1% (n = 419). From gestation Day 34 to the 90th day of pregnancy, there were 183 abortions (6.6% of all pregnancies) (23 in Neospora positive animals). After 90 days of pregnancy, the number of abortions was 146 (5.3%); 126 occurring during the second and 20 during the third trimester of pregnancy (105 in Neospora positive animals). Multiple logistic regression analyses were performed on data from each animal using abortion before or after 90 days of pregnancy as the dependent variable, and Neospora positivity, herd, pregnancy season, and parity (parous or nonparous) as independent factors. No significant effects of Neospora positivity and herd were found on the abortion rate before 90 days of pregnancy. Based on the odds ratio, the abortion rate was 4 times higher (P < 0.0001) in animals that became pregnant in the warm than in the cool period, and 3.7 times higher (P < 0.0001) in parous than in nonparous animals. Neospora positivity was the only variable included in the logistic regression model for abortions occurring after 90 days of pregnancy. Seropositivity in an animal increased the probability of abortion by an odds ratio of 18.9 (P < 0.0001; 95% confidence interval 12.9-27.8). Season, parity, and herd showed no effect. The results of the present study suggest that chronic N. caninum infection prior to pregnancy appears not to affect the early fetal period, but does have a significant abortive effect after 90 days of gestation.     

Ability of induced corpora lutea to maintain pregnancy from the third month of gestation to term in cattle

The local relationship between the pregnant uterine horn and the CL during maternal recognition of pregnancy is well-documented. It continues beyond that time; pregnancies were maintained in lutectomized cows when CL were induced on the ovary ipsilateral, but not contralateral, to the uterine horn of pregnancy during Days 28-53. This study evaluated factors affecting maintenance of pregnancy by CL induced after Day 53, in lutectomized cows that had received exogenous progesterone from Day 29 to 15 days after induction of a CL. Twenty-four suckled beef cows were lutectomized on Day 29 of gestation; pregnancy was maintained with progesterone from two controlled internal drug releasing (CIDR) inserts, exchanged every 5 days. Beginning on Day 53, ovaries and viability of pregnancy were evaluated by ultrasonography every 5 days. When a follicle >or=10 mm in diameter was present ipsilateral to the fetus, each cow received 1,000 IU of hCG. Following induction of a CL (20 of 24), progesterone was reduced to a single CIDR for 5 days, then removed. Retention of pregnancy was confirmed by rectal palpation and calving. Cows with induced CL maintained pregnancy to term, including four with the CL contralateral to the fetus. Three cows failed to form normal CL by Day 98 and lost pregnancy after removal of exogenous progesterone. One cow that did not respond to hCG lost pregnancy during exogenous progesterone. In conclusion, CL induced after Day 53 maintained pregnancy to term, even when induced contralateral to the pregnant uterine horn.     

Endocrine profiles of somatic nuclear transfer-derived pregnancies in dairy cattle

In cattle, several hormones and proteins are necessary for maintenance of a normal pregnancy that will result in a viable calf. Deviation from the normal cascade or expected profile of reproductive hormones and proteins may be associated with impairment of somatic nuclear transfer-derived pregnancies and the high rate of fetal loss. The objectives of this study were to characterize maternal plasma concentrations of pregnancy-specific protein B (PSPB), progesterone (P4), estrone sulphate (E₁S), and estradiol (E2) during the last two-thirds of pregnancy (cloned calves), and to determine associations with gestational abnormalities. Cows with cloned fetuses, produced by either commercial (N = 16) or zona-free (N = 4) cloning techniques, were compared with pregnant animals derived from traditional embryo transfer (N = 6) or AI (N = 6), at various stages of gestation (Days 80, 120, 150, 180, 210, and 240; Day 0 = estrus). Fetal well-being was monitored with ultrasonography throughout gestation. At Day 80, progesterone concentration was lower (P < 0.0001) in nuclear transfer (NT) recipients than in control groups. Mean estrone sulphate concentrations did not vary significantly between NT and control groups. At Day 150, pregnancy-specific protein B concentrations were elevated (P < 0.002) in NT cows. Estradiol concentration was higher in NT recipients than control cows throughout the study period.     

Peripheral plasma progesterone concentration in zebu (Bos indicus) cows during pregnancy

Plasma progesterone concentrations were determined weekly during gestation averaging 283 +/- 2 d in Ethiopian zebu (Bos indicus) cows. Mean progesterone levels increased from 0.2 +/- 0.1 ng/ml at oestrus (service) to 3.1 +/- 1.6 ng/ml on d 7 and 8.1 +/- 2.1 ng/ml on d 21. Progesterone levels remained elevated throughout pregnancy. Hormone concentration differed significantly between cows (P less than 0.001) and with the wk of pregnancy (P less than 0.05); it tended to be higher during the last trimester of pregnancy. Mean levels dropped sharply to below 1 ng/ml during the last wk of pregnancy with considerable variation (C.V. = 39 to 63%) among cows. These data indicate that pregnancy in Ethiopian zebu cows can be reliably diagnosed by determining circulatory plasma progesterone levels.     

Low peripheral progesterone and late embryonic/early fetal loss in suckled beef and lactating dairy cows

Pregnancy failure during placentation in lactating dairy cows was associated with low concentrations of serum progesterone. Beef cows have greater serum progesterone and less pregnancy failure. Experiment 1 determined that reduction of serum progesterone affected late embryonic/early fetal loss in suckled beef cows. Cows (n = 40) received progesterone from two new or used controlled internal drug releasing devices, replaced every 5 d, beginning on Day 28 of gestation (mating = Day 0); CL were enucleated on Day 29. Retention of pregnancy was 77% in treated cows and 97% in 78 control cows (P < 0.05). Experiment 2 determined how pregnant, lactating dairy cows with high or low progesterone concentrations during Days 28-34 differed in luteal function or in serum progesterone during replacement therapy. Luteal tissue from such cows was assayed for progesterone and expression of mRNA for genes of endothelin and prostaglandin (PG) systems. Secretion of progesterone and prostaglandins by dispersed luteal cells was determined during incubation with LH, endothelin-1, or arachidonic acid. Neither luteal progesterone nor mRNAs for endothelin or prostaglandin systems differed. Endothelin-1 inhibited secretion of progesterone more (P < 0.05) in luteal cells from cows with low versus high serum progesterone, when incubated with arachidonic acid. Secretion of prostaglandin F₂α was increased and that of 6-keto-PGF₁α decreased by endothelin-1 in vitro. Serum progesterone during replacement was lower (P < 0.05) for cows with low than high serum progesterone at lutectomy. Thus, clearance, more than luteal production, determined peripheral progesterone in pregnant, lactating dairy cows.     

Evaluation of different techniques for pregnancy diagnosis in sheep

Fifty Corriedale ewes were used in this study to evaluate pregnancy diagnosis in sheep. Ewes were bred under a pen mating system and pregnancy diagnosis was initiated from day 15 post-mating, applying the diagnostic techniques of trans-abdominal real-time B-mode ultrasonography, Preg-alert (A-mode ultrasonography), the Doppler ultrasonic fetal pulse detector or the plasma progesterone concentration assay (EIA). These tests were repeated fortnightly on all the ewes until the onset of lambing. The accuracy of trans-abdominal real-time B-mode ultrasonography (68%) at days 15–30 of pregnancy increased to 100% by days 61–75 and remained constant until lambing. The accuracy of the Preg-alert (56%) diagnosis at days 31–45 increased to 94% by days 91–105 of gestation and then decreased to 82% from days 136 of gestation to lambing. The accuracy of both the Doppler ultrasound (56%) at days 31–45 and plasma progesterone assay (98%) at days 15–30 of gestation increased to 100% at days 76–90 and 46–60 of gestation, respectively and remained constant until parturition. The mean plasma progesterone concentration at days 0–6 (1.41 ± 0.21 ng/ml) increased to 4.0 ± 0.87 ng/ml at days 16–30 (days 18.23 ± 0.78) post-mating. Animals returning to estrus recorded less than 1 ng/ml at days 18.23 ± 0.78 post-mating. The accuracy of both the B-mode ultrasonic technique (78%) and plasma progesterone assay (98%) was significantly higher (P < 0.05) than the accuracy obtained with the A-mode and Doppler ultrasound (both 56%) at days 31–45 of gestation. The study concluded that real-time B-mode ultrasonography is the earliest, most accurate, safest, fastest and most economical method of pregnancy diagnosis in sheep at farm level. The A-mode and Doppler methods can also be used under field conditions, where sophisticated laboratory facilities are not available. Plasma progesterone assays (EIA) can be used as a means of early pregnancy diagnosis in organized sheep farms with fair accuracy.     

Pregnancy patterns during the early fetal period in high producing dairy cows treated with GnRH or progesterone

In order to explore pregnancy patterns in high producing dairy cows treated with GnRH or progesterone at pregnancy diagnosis (Days 28-34), two consecutive experiments were designed. In Experiment 1, cows bearing a single embryo were randomly assigned to a PRID (n = 40; cows fitted with a progesterone releasing intra-vaginal device for 28 days), GnRH (n = 40; cows receiving GnRH) or Control (n = 26; untreated cows) group. PRID treatment led to a rise in plasma progesterone concentrations in the 7 days following the onset of treatment compared to the other two groups. In Experiment 2, in which we also examined twin pregnancies, animals were randomly assigned to PRID (n = 312) or GnRH (n = 294) treatment groups. Treatments were the same as described for Experiment 1. Logistic regression procedures revealed that in cows with a single corpus luteum, the probability of pregnancy loss between the first (Days 28-34) and second (Days 65-62) pregnancy diagnosis decreased by a factor of 0.51 in the PRID group compared to the GnRH group. However, in cows with two or more corpora lutea, PRID treatment increased the likelihood of pregnancy loss by a factor of three, compared to GnRH treatment. In cows carrying twins, the conceptus reduction rate was higher (P = 0.02) for the GnRH (36%) than for the PRID (16.4%) group. Formation of a new corpus luteum was recorded in 17.7% of cows in the GnRH group. Our results indicate that compared to GnRH treatment, progesterone treatment given at pregnancy diagnosis in high producing dairy cows, reduced by a factor of 0.51 and increased by a factor of 3 the probability of pregnancy loss in cows with a single or with two or more corpora lutea, respectively, and reduced the conceptus reduction rate in cows carrying twins. The practical implications of our findings are that in herds with a high incidence of early fetal loss of a non-infectious nature, treatment at the time of pregnancy diagnosis with PRID in cows with one corpus luteum and with GnRH in cows with two or more corpora lutea should offer considerable benefits.     

Factors affecting retention of early pregnancy in dairy cattle

Potential factors affecting retention of pregnancy during weeks 5-9 of gestation were studied in dairy cows and heifers (N = 211) on two farms. Cows were examined by ultrasonography for presence of a viable embryo, and sizes of the corpus luteum (CL) and of follicles > or = 5mm were recorded. Blood samples were taken at each examination and assayed for progesterone and estradiol. Overall pregnancy loss was 11.4%. Cows with two CL did not have greater concentrations of progesterone than cows with one CL and they retained fewer pregnancies (P < 0.01; 73% versus 91%). Pregnancy retention was associated positively with concentrations of progesterone and estradiol during week 5 (P < 0.05). Embryos that were lost apparently died before CL regression. Retention of pregnancy declined in cows with high body condition and as age of the cow increased. Pregnancy retention was lower in cows bred to one of four frequently-used service sires (P < 0.05). Days postpartum, milk production, parity, service number, inseminator, synchronization of estrus, diameter of follicles and size of CL did not affect pregnancy retention. In conclusion, retention of pregnancy during placentation varied with concentrations of progesterone and estradiol, age of cow, body condition and service sire.     

Concentrations of progesterone in milk as a monitor of early pregnancy diagnosis in dairy cows

The objectives of the experiment were to evaluate the efficacy of using progesterone concentrations in milk and palpation per rectum on days 21 or 22 postbreeding to estimate pregnancy and evaluate management practices; and to investigate physiological occurrences leading to incorrect diagnosis of pregnancy when serial samples of milk were collected. Of particular interest were indications of early embyronic death and insemination of cows not in estrus. Milk samples were collected at the afternoon milking of days 0 or 1 (day 0 = day of estrus), 9 or 10, 21 or 22 and 27 or 28 following breeding in 200 lactating dairy cows. Tentative diagnosis of pregnancy was made based on concentrations of progesterone in milk on days 21 and 22 alone and on days 21 or 22 and 27 or 28. In addition all cows were palpated per rectum on days 21 or 22 postbreeding and a tentative pregnancy diagnosis was made. Pregnancy was confirmed by examination of the genital tract per rectum between 35 and 50 days after breeding. Values of 4 ng/ml or greater and/or the presence of a mature corpus luteum were considered positive signs of pregnancy. Progesterone in milk ranged from 0.1 to 18 ng/ml. On days 0 or 1, 9 or 10, 21 or 22 and 27 or 28 concentrations of progesterone in milk averaged 1.5 +/- 0.3, 11.1 +/- 0.5, 12.0 +/- 0.4 12.5 +/- 0.5 ng/ml for pregnant cows. Corresponding samples from nonpregnant cows averaged 1.2 +/- 0.2, 10.3 +/- 0.4, 3.0 +/- 0.4, 6.8 +/- 0.6 ng/ml, respectively. Ninety-six and 104 cows were classified as pregnant and nonpregnant on days 21 or 22 as compared to 78 and 118 cows diagnosed as pregnant and nonpregnant on days 21 or 22 and 27 or 28 combined. Pregnancy detection by progesterone in milk on days 21 or 22 with pregnancy determined via rectal palpation 35 to 50 days postbreeding was 77 and 100% accurate for positive and negative diagnosis, respectively. The percent agreement using progesterone in milk on days 21 or 22 and 27 or 28 combined was 95 and 100%, respectively, for positive and negative diagnosis. Diagnosis based on rectal palpation 21 or 22 days postbreeding was 63 92 (69%) and 76 88 (87%) for pregnant and nonpregnant cows, respectively. Ten of the 200 cows had progesterone concentratins in milk of > 4 ng/ml at the time of breeding. Six of these cows were pregnant from a previous insemination. The other four cows were nonpregnant and were inseminated during the luteal phase of the cycle. In conclusion, measurement of progesterone in milk is a useful tool in early detection of pregnant and nonpregnant cows and may be useful in detecting reproductive problems in a dairy herd. It will probably be most useful when used in combination with later pregnancy diagnosis per rectum .