Cattle fetal sex determination by polymerase chain reaction using DNA isolated from maternal plasma

The objective of this study was to evaluate the use of polymerase chain reaction analysis (PCR) of fetal cells/DNA in the maternal plasma of pregnant cows to determine the sex of the fetus. Plasma was harvested from 35 cows of mixed genotype at different stages of pregnancy ranging from 5 to 35 weeks. A male calf and a heifer calf provided the control samples. Fetal sex was determined by amplification of Y-specific sequences. For the 35 cows, the fetal sex predicted by this technique was in accordance with the sex of the calf at birth in 88.6% of cases. The agreement between predicted and observed fetal sex was less for cows with a gestational length of 35-48 days (63.6%). Regression analysis showed that there was a strong relationship between the probability of correctly predicting fetal sex and the stage of gestation. It was estimated that the test performed at 43.8 days post fertilization would have 95% accuracy, increasing to 99% accuracy for testing at 48.4 days and 99.9% accuracy for tests at 55.0 days or later. It was concluded that PCR analysis of fetal cells in maternal plasma can be used to predict successfully the sex of the fetus in cattle.     

Bovine fetal microchimerism in normal and embryo transfer pregnancies and its implications for biotechnology applications in cattle

Fetal cells and DNA have been detected in the maternal circulation during and after pregnancy in a few mammalian species. The incidence of similar microchimerism in cattle could have repercussion for the application of modern biotechnologies such as the transfer of transgenic embryos. To determine if feto-maternal leakage can occur in pregnant cows, we have analyzed maternal blood samples for the presence of fetal DNA during gestation and post-partum periods. Y chromosome-specific DNA was detected in up to 73% of blood samples from naturally mated heifers carrying conventional bull calves and a transgene-specific sequence in up to 50% of recipient cows carrying transgenic fetuses. These findings document for the first time that transplacental leakage of fetal DNA into the maternal circulation can occur in cattle despite the epitheliochorial placenta of ruminants, with potential implications for the utilization of recipient cows in the food chain.     

Determination of bovine fetal sex by PCR using fetal fluid aspirated by transvaginal ultrasound-guided amniocentesis

Fetal sex can be determined by the polymerase chain reaction (PCR) using cells from fetal fluid collected by transvaginal ultrasound-guided amniocentesis. A total of 35 aspirates from 30 cows, 15 Holsteins and 15 Japanese Blacks at 59 to 250 d of pregnancy were used. Five cows were aspirated twice at a 10-d interval. A 5.0 MHz convex array transducer connected to a scanner was inserted into the vagina under caudal epidural anesthesia. The transducer was equipped with a 65-cm long, 21-g needle within the probe carrier. A bovine male-specific primer and a bovine gender-neutral primer were used. Fetal fluid was obtained from all except 2 cows in early pregnancy. Five animals aborted within 1 wk following aspiration. A total of 33 samples, 29 of amniotic fluid and 4 of allantoic fluid, was subjected to PCR analysis. Fetal gender was verified in 31 33 samples (18 females and 13 males). Gender was also determined by gross examination of external genitalia of offspring after calving or abortion. Fetal gender was correctly identified by PCR analysis of aspirated fetal fluid in 16 16 females and in 13 15 males. Transvaginal ultrasound-guided amniocentesis followed by PCR analysis of aspirated cell DNA can be used accurately to determine fetal sex in cows at 70 to 100 d of gestation. The procedure requires considerable skill and is not without some risk to fetal viability.     

Bovine maternal,fetal and neonatal responses to bovine viral diarrhea virus infections

Due to the affinity of BVDV for the fetus and for cells of lymphatic organs of infected cattle, reproductive failure or immunosuppression, respectively, are likely consequences of BVDV infections of susceptible cattle. Infection of susceptible pregnant cattle with noncytopathic (ncp) BVDV results in transplacental infection with induction of maternal and fetal innate and adaptive immune responses. Differences in maternal innate and adaptive immune responses are evident in late gestation between cows carrying fetuses persistently-infected (PI) with BVDV and cows with fetuses transiently-infected with BVDV. Fetal innate and adaptive immune responses to ncp BVDV infection are defined by fetal age and developmental stage of the fetal immune system. Since a functional fetal adaptive immune response does not occur in the early fetus, immunotolerance to ncp BVDV is established, virus replicates unrestricted in fetal tissues and calves are born immunotolerant and PI with the virus. In the last trimester of gestation, the fetal immune system is adequately developed to respond in an efficacious manner, most commonly resulting in the birth of a clinically normal calf with pre-colostral antibodies. Immunosuppression due to postnatal acute ncp BVDV infections of susceptible calves may contribute to the occurrence and severity of multi-factorial respiratory tract and enteric diseases.     

Noninvasive Fetal Sex Determination by Real-Time PCR and TaqMan Probes

Background:Noninvasive fetal sex determination by analyzing Y chromosome-specific sequences is very useful in the management of cases related to sex-linked genetic diseases. The aim of this study was to establish a non-invasive fetal sex determination test using Real-Time PCR and specific probes.Methods:The study was a prospective observational cohort study conducted from August 2018 to September 2019. Venous blood samples were collected from 25 Iranian pregnant women at weeks 7 to 25 of gestation. Cell-free DNA (cfDNA) was isolated from the plasma of samples and fetal sex was determined by SRY gene analysis using the Real-Time PCR technique. In the absence of SRY detection, the presence of fetal DNA was investigated using cfDNA treated with BstUI enzyme and PCR for the epigenetic marker RASSF1A.Results:Of the total samples analyzed, 48% were male and 52% female. The RASSF1A assay performed on SRY negative cases also confirmed the presence of cell-free fetal DNA. Genotype results were in full agreement with neonate gender, and the accuracy of noninvasive fetal sex determination was 100%.Conclusion:Fetal sex determination using the strategy applied in this study is noninvasive and highly accurate and can be exploited in the management of sex-linked genetic diseases.Key Words: Cell-free fetal DNA, Fetal sex determination, Noninvasive prenatal diagnosis, Sex-linked genetic diseases, SRY     

Androgens in the bovine fetus and dam (38567).

Umbilical arterial and venous blood, and fetal testes were taken from 38 bovine fetuses at 90, 180 or 260 days of gestation. Concurrently blood also was taken from the jugular, and from the uterine artery and vein of the dams. Testosterone and androstenedione were determined by radioimmunoassays. Fetal testicular homogenates had 0.96 and 0.35 mug/g of testosterone and 0.39 and 0.50 mug/g of androstenedione at 180 and 260 days of gestation, respectively. Males had five to tenfold more serum testosterone and about twofold more androstenedione than female fetuses at each trimester of gestation. Male fetal blood testosterone decreased (P less than 0.01) from 2.7 to 0.3 ng/ml between 90 and 260 days of gestation. But, maternal testosterone and androstenedione increased (P less than 0.05) during gestation in cows with males, but not in cows with female fetuses. Testosterone was higher (P less 0.05) in cows carrying males than in cows with female fetuses. Androstenedione was higher in blood leaving the placenta on both the maternal and on the vetal sides suggesting placental synthesis of androstenedione.     

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.     

Influence of sire, sex of fetus and type of pregnancy on conceptus development

Estrus was synchronized in 93 crossbred cows 3 to 7 yr old which were then superovulated with 10 mg FSH-P and bred artificially to either a Jersey or Charolais bull. Females pregnant to the Jersey sire were slaughtered at 95, 180 or 250 d of gestation; low pregnancy rate from the Charolais sire resulted in enough dams for slaughter and valid comparisons at 95 d only. Conceptus tissue and organ weights and dam carcass weights and measurements were collected at slaughter. At 95 d of gestation, sire differences were not significant; type of pregnancy (single vs multiple) had significant effects on placentome and membrane weight; fetus sex had highly significant effects on fetus weight and crown-rump length. Fetal brain and heart weights showed little effect of sire, fetus sex or type of pregnancy, indicating these early differentiating tissues are highly competitive for available nutrients for growth. Fetal data at 180 and 250 d of gestation showed highly significant effects of gestation stage on placentome, membrane and fetus weights, crown-rump and metatarsal lengths, and liver, kidney, heart and total brain weights. Type of pregnancy had greater effects on the conceptus traits than did fetus sex at these stages. Nonorthogonal analyses of data from the Jersey-sired pregnancies indicated growth of fetuses in multiple pregnancies was suppressed, and the reduction in fetus weight became more pronounced as gestation progressed. Correlations among conceptus traits and dam carcass data showed different relationships within single and multiple pregnancies. We suggest that the genetic contribution for fetal growth has a positive effect on fetal size throughout gestation while the importance of metabolite availability may change during gestation depending on the genetic growth potential of the fetus and on whether the pregnancy is single or multiple. We speculate that these fetus-dam relationships involve systems controlling fetus growth that arise from the fetus genetic growth potential interacting with positive and negative dam effects that are linked or related to the carcass skeletal-fat-muscle endpoints studied.     

Fetal DNA detection in maternal plasma throughout gestation

The presence of fetal DNA in maternal plasma may represent a source of genetic material which can be obtained noninvasively. We wanted to assess whether fetal DNA is detectable in all pregnant women, to define the range and distribution of fetal DNA concentration at different gestational ages, to identify the optimal period to obtain a maternal blood sample yielding an adequate amount of fetal DNA for prenatal diagnosis, and to evaluate accuracy and predictive values of this approach. This information is crucial to develop safe and reliable non-invasive genetic testing in early pregnancy and monitoring of pregnancy complications in late gestation. Fetal DNA quantification in maternal plasma was carried out by real-time PCR on the SRY gene in male-bearing pregnancies to distinguish between maternal and fetal DNA. A cohort of 1,837 pregnant women was investigated. Fetal DNA could be detected from the sixth week and could be retrieved at any gestational week. No false-positive results were obtained in 163 women with previous embryo loss or previous male babies. Fetal DNA analysis performed blindly on a subset of 464 women displayed 99.4, 97.8 and 100% accuracy in fetal gender determination during the first, second, and third trimester of pregnancy, respectively. No SRY amplification was obtained in seven out of the 246 (2.8%) male-bearing pregnancies. Fetal DNA from maternal plasma seems to be an adequate and reliable source of genetic material for a noninvasive prenatal diagnostic approach.     

Ultrasonic determination of fetal gender in horses and cattle under farm conditions

Accuracy of transrectal ultrasonic diagnosis of fetal gender by identifying and locating the genital tubercle was assessed in 85 horses and 102 dairy heifers and cows. Examinations were made once (horses and cattle) under farm conditions (cattle) on approximately Days 50 to 100. Definite diagnosis was made by removal of fetuses (horses) or after calving or abortion (cattle). In both species and both parities, accuracy was 100% (109 109 totaled over both species) when the certainty level recorded at the time of examination was 95 or 99%. At the lower levels of certainty (65 to 80%, 85 to 90%), the accuracy was 89% (57 64 totaled over both species). A diagnosis was not obtainable in 10 horses (12%) and 5 cattle (5%); this was especially due to inadequate viewing of the fetus when the fetus was beyond Day 64. The time from completion of evacuation of the rectum to diagnosis of gender averaged 1 minute 17 seconds in horses (range, 15 seconds to 3 minutes 55 seconds). The corresponding figure in cattle was 1 minute 53 seconds (range, 16 seconds to 8 minutes 30 seconds).     

Influence of gestation on jugular plasma thyroxine levels in Murrah buffalo and Karan Swiss cows

Plasma samples from 106 pregnant Karan Swiss (Brown Swiss x Sahiwal) cows and 108 Murrah buffalo were tested for thyroxine (T(4)) levels to determine the relationship between the hormonal changes and advancing pregnancy in the two species. All samples were collected within 2 months (January and February) to avoid seasonal interference on T(4) levels. In pregnant cows, the concentration of T(4) increased sharply during the first trimester, reaching a peak at the third month of gestation followed by a gradual decline until the last month of pregnancy. In pregnant buffalo, peripheral plasma T(4) levels fluctuated slightly throughout pregnancy without exhibiting a specific trend. Statistical analysis revealed that the magnitude of T(4) levels was significantly lower in buffalo (P < 0.01) than in the cows throughout pregnancy and that the hormonal patterns of the two species were significantly different (P < 0.05) during gestation. It was hypothesized from this study that T(4) requirements for the fetal buffalo calf may be lower than that for the fetal cattle calf since the buffalo gestation period is a month longer and the metabolic rate lower vis-a-vis the cow.     

Use of real-time ultrasound to identify multiple fetuses in beef cattle

To examine the feasibility of producing multiple births in beef cattle by means of superovulation, realtime ultrasound was used to identify cows carrying multiple fetuses. Three replicates of the superovulation experiment were conducted with groups of 52, 25 and 89 purebred Angus cows, respectively. Both uterine horns of cows from Replicate 1 were examined via the rectum using ultrasound at averages of 43 (range = 30 to 68 days), 51 (range = 38 to 76 days) and 126 (range = 113 to 151 days) days after AI. Cows in Replicate 2 were examined in the same fashion at averages of 55 and 97 days post insemination. In Replicate 3 the number of fetuses was estimated on a single date (average number of days post insemination = 49 days). Across the 3 replicates, the number of fetuses was most accurately assessed at an average of between 49 and 55 days post insemination. In most instances for which comparability between ultrasound estimations and calving results was low, the lack of correspondence was likely due to embryo mortality in cows identified as carrying multiple fetuses. For all 3 replicates combined, only 1 cow diagnosed with a single fetus produced multiple calves at birth when the diagnoses were conducted at 49 to 55 days post insemination. Real-time ultrasound can be used to accurately identify open cows and to differentiate between cows carrying single or multiple fetuses.     

Equine fetal sex determination using circulating cell-free fetal DNA (ccffDNA)

In this study, polymerase chain reaction (PCR) reamplification of the first PCR product (2nd-PCR) and a qPCR assay were used to detect the sex determining region Y (SRY) gene from circulating cell-free fetal DNA (ccffDNA) in blood plasma of pregnant mares to determine fetal sex. The ccffDNA was isolated from plasma of 20 Thoroughbred mares (5-13 y old) in the final 3 mo of pregnancy (fetal sex was verified after foaling). For controls, plasma from two non-pregnant mares and two virgin mares were used, in addition to the non-template control. The 182 bp nucleotide sequence corresponding to the SRY-PCR product was confirmed by DNA sequencing. Based on SRY/PCR, 8 of 11 male and 9 of 9 female fetuses were correctly identified, resulting in a sensitivity of 72.7% (for male fetuses) and an overall accuracy of 85%. Furthermore, using SRY/2nd-PCR and qPCR techniques, sensitivity and accuracy were 90.9 and 95%, respectively. In conclusion, this study is apparently the first report of fetal sex determination in mares using ccffDNA.     

SRY sequence in maternal plasma: Implications for non-invasive prenatal diagnosis: First report from India

AIM:

The presence of circulatory cell-free fetal DNA in maternal plasma has found new applications in non-invasive risk-free prenatal diagnosis.

MATERIALS AND METHODS:

We made use of a size separation approach along with real time polymerase chain reaction (PCR) to evaluate the use of fetal DNA in the detection of the sex of the fetus. Cell-free fetal DNA was isolated from the plasma of 30 women (10–20 weeks gestation) using a size separation approach. We made use of Taq Man Chemistry and real time PCR using primers and probes for GAPDH and SRY.

RESULTS:

Only 24 cases could be studied as there was no amplification in six cases. Fetal sex was accurately determined in all of the 24 cases wherein 19 women were carrying male fetuses and five women were carrying female fetuses. An increase in the amount of fetal DNA was observed with an increase in the gestational age.

CONCLUSIONS:

Real time PCR analysis is a highly sensitive and accurate tool for non-invasive prenatal diagnosis, allowing detection of the sex of the fetus as early as 10 weeks of gestation. Non-invasive prenatal diagnosis eliminates the risk of fetal loss associated with the invasive procedure.     

Melatonin supplementation alters uteroplacental hemodynamics and fetal development in an ovine model of intrauterine growth restriction

Using a mid- to late-gestation ovine model of intrauterine growth restriction (IUGR), we examined uteroplacental blood flow and fetal growth during melatonin supplementation as a 2 × 2 factorial design. At day 50 of gestation, 32 ewes were supplemented with 5 mg of melatonin (MEL) or no melatonin (CON) and were allocated to receive 100% [adequate; (ADQ)] or 60% [restricted (RES)] of nutrient requirements until day 130 of gestation. Umbilical artery blood flow was increased from day 60 to day 110 of gestation in MEL vs. CON dams, while umbilical artery blood flow was decreased from day 80 to day 110 of gestation in RES vs. ADQ dams. At day 130 of gestation, uteroplacental hemodynamics, measured under general anesthesia, and fetal growth were evaluated. Uterine artery blood flow was decreased in RES vs. ADQ dams, while melatonin supplementation did not affect uterine artery blood flow. Total placentome weight and placentome number were not different between treatment groups. Fetal weight was decreased by nutrient restriction. Abdominal girth and ponderal index were increased in fetuses from MEL-ADQ dams vs. all other groups. Fetal biparietal distance was decreased in CON-RES vs. CON-ADQ dams, while melatonin supplementation rescued fetal biparietal distance. Fetal kidney length and width were increased by maternal melatonin treatment. Fetal cardiomyocyte area was altered by both maternal melatonin treatment and nutritional plane. In summary, melatonin may negate the consequences of IUGR during specific abnormalities in umbilical blood flow as long as sufficient uterine blood perfusion is maintained during pregnancy.     

A method to reduce the invasiveness of fetal catheterization in the cow

Surgical intervention in general anesthesia (GA) of the cow in late gestation is a stressful condition for both mother and fetus, potentially leading to premature delivery or fetal death. The present study hypothesized that fetal catheterization at days 246–253 (90% of gestation) is done with less physical and metabolic stress for the mother and fetus, when the surgery is performed on a standing cow and local anesthesia (LA) rather than on a recumbent cow in general anesthesia. Fetal and uterine maternal intra-vascular catheters were implanted during general anesthesia (GA, n=24) or local analgesia (LA, n=7). Blood gases and metabolite levels in the fetal calves and their mothers were measured during surgery and for 5 days post-operatively. During surgery, venous blood pH was higher (7.44±0.01 versus 7.39±0.01, P<0.05) and hemoglobin and oxygen contents lower in LA cows compared with GA cows (9.3±0.3 mg/dl versus 11.8±0.2 mg/dl, P<0.001 and 10.1±0.3 ml/dl versus 12.6±0.6 ml/dl, P<0.05). The differences between the two groups of fetuses reflected those of their dams in that LA fetuses showed lower arterial oxygen pressure (18.3±1.4 mmHg versus 24.8±1.4 mmHg, P<0.05) and hemoglobin (7.81±0.30 mg/dl versus 9.22±0.21 mg/dl P<0.01) and furthermore, they also showed higher blood glucose (2.4±0.2 mM versus 1.4±0.1 mM, P<0.01). During the 5 days post-surgery, 10 GA fetuses (42%) and 1 LA fetus (14%) died in utero. Bacterial contamination was implicated in six of the GA deaths and in the one LA death. In the dams with surviving calves, differences in hemoglobin (9.49±0.21 mg/dl versus 11.17±0.23 mg/dl P<0.001) and O₂ct (10.9±0.3 ml/dl versus 12.5±0.5 ml/dl, P<0.05) were still present, and in addition, blood glucose was higher in LA versus GA cows (4.3±0.2 mM versus 3.8±0.1 mM, P<0.05). The choice of surgical method did not affect post-surgery blood chemistry in the surviving fetuses, except that the higher blood glucose in the LA fetuses at surgery tended to be maintained also post-operatively (2.0±0.2 mM versus 1.5±0.1 mM, P=0.07). The observed differences in blood chemistry parameters between the two methods of surgery and possibly in the fetal death may be explained by differences in catheterization method and the associated differences in physical and metabolic stress during and after surgery. Thus, surgery upon a standing cow in local anesthesia should be considered as an alternative to surgery in universal anesthesia for fetal catheterization in the cow in late gestation.     

Fetal cells in the maternal circulation during first trimester in pregnancies

To investigate the presence of fetal cells in the maternal circulation during early pregnancy, the polymerase chain reaction was used to test the presence of human Y chromosome-specific ZFY and SRY gene DNA sequences in maternal peripheral blood specimens from 19 women carrying male fetuses and 12 women carrying female fetuses. The presence of fetal cells was suggested as early as 6 weeks gestation in 1 of the 19 women bearing male fetuses. Fetal cells were present in the maternal circulation of 15 of the 19 women by 9 weeks gestation, and in only 1 of the 19 were fetal cells not detected until the 12th week after conception. These results suggest that identification of fetal cells in the maternal circulation is possible with a properly designed and executed polymerase chain reaction. However, there was considerable variation with respect to when these fetal cells first became detectable during pregnancy. These fetal cells are potentially a valuable source of material for biochemical and genetic studies of the fetuses.     

Ultrasonographic fetometry and determination of fetal sex in buffaloes (Bubalus bubalis)

The aim of the current study was to establish ultrasonic biometric threshold of different fetal parts in buffaloes and to evaluate the feasibility and accuracy of ultrasonic fetal sex determination. Serial ultrasonographic examinations were carried out on twelve pregnant buffalo-cows, during which fetal parts were measured, and fetal sex and presentation were determined. The obtained results revealed that embryo and amniotic vesicle (AV) were detected by the forth and fifth week of pregnancy, respectively. Organization was observed by the seventh week, while ossification was indicated between the eighth and 10th week. High correlations were found between different studied parameters and gestational age, where the highest correlation was found with the crown-rump length (CRL) and amniotic vesicle diameter (AVD) at the early-gestation; the biparietal diameter (BPD) at the mid-gestation; and the eyeball diameter (EBD) at the mid- and late-gestation. The results also revealed that the best window for fetal sexing was found between the 10th and 18th week of gestations, with an overall accuracy of 97.1%. The final polarity with all fetuses in anterior presentation was adopted by the 30th week. In conclusion, the overall data indicated the feasibility and value of ultrasonographic fetometry in buffaloes for evaluation of fetal development, estimation of gestational age and determination of fetal sex.     

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.     

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.