Evaluation of a Commercial Pregnancy Test Using Blood or Plasma Samples in High-Producing Dairy Cows.

This study evaluated a commercial pregnancy-associated glycoproteins (PAGs)-based pregnancy test using whole blood or plasma samples during early pregnancy (28-55 days of gestation) in high-producing dairy cows. Transrectal ultrasonography was used as the gold standard method. The study population constituted of 284 cows. False positive diagnoses were recorded from Day 60 to 89 and from Day 60 to 99 postpartum in blood and plasma samples, respectively. In early pregnancy screening, correct positive diagnoses were recorded in 75% and 100% of blood and plasma samples, respectively. High milk production was associated with negative results in blood samples and with the lowest test line intensity in plasma samples. False positive or negative diagnoses were recorded in 0% of both types of samples in cows previously diagnosed as pregnant and showing signs of estrus. In conclusion, the use of plasma was more effective than the use of blood in early pregnancy diagnosis. In cows previously diagnosed as pregnant and showing signs of estrus, both types of samples showed the same results. Because of large individual variations, normal single pregnancies could not be differentiated from twin pregnancies, from pregnancies with a recently dead conceptus, or from pregnancies that experienced subsequent pregnancy loss.

Negative photoperiod induces an increase in the number of ovulations in dairy cattle.

This study sought to examine the impact of negative photoperiod on the incidence of multiple ovulations and pregnancies in dairy cattle. The study population consisted of 5,373 pregnant cows in their third or greater lactation that experienced their first post-partum pregnancy after spontaneous estrus. The positive photoperiod (increasing day-length) extends from December 22 to June 21, whereas the negative photoperiod (decreasing day-length) extends from June 22 to December 21. The odds ratios (ORs) for multiple ovulations and pregnancies in cows that became pregnant during the negative photoperiod and the remaining cows that became pregnant during the positive photoperiod were 1.4 and 1.3 (P < 0.0001), respectively. The ORs for cows that became pregnant ≥ 90 days in milk and the remaining cows that became pregnant < 90 days in milk were 4.3 and 4.1 (P < 0.0001), respectively. No significant differences were detected in the monthly rates of multiple ovulations or pregnancies during positive and negative photoperiods. Thus, the present study demonstrates that the ovarian function in cows is related to changes in day-length, with decreasing day-length being associated with greater multiple ovulation and pregnancy rates. The present study also shows that positive and negative photoperiods exhibit different trends. The results of this study are consistent with a growing body of work demonstrating the effects of photoperiod patterns on the reproductive physiology of cows, with clear implications for twin pregnancy prevention.

Effect of Genetic and Environmental Factors on Twin Pregnancy in Primiparous Dairy Cows.

Twin pregnancies are highly undesirable in dairy cattle; they compromise the health and wellbeing of a cow and dramatically impair the farm economy. Recently, a genomic prediction for twin pregnancies has been developed. The objective of this study was to assess environmental and management risk factors affecting the incidence of twin pregnancies in high-producing dairy cows in their first lactation, with a special emphasis placed on the genomic prediction values of twin pregnancy. Our study population of primiparous cows proved valuable in identifying factors other than genomic predictive values that influence twin pregnancy rates. The odds ratio for twin pregnancies was 0.85 (p < 0.0001) for each unit of a prediction value increase, 3.5 (p = 0.023) for cows becoming pregnant during the negative photoperiod, and 0.33 (p = 0.016) for cows producing ≥42 kg of milk at AI, compared with the remaining cows who produced <42 kg of milk. As a general conclusion, the practical implication of our findings is that genomic prediction values can identify the risk of twin pregnancy at a herd level. Given the cumulative effect of genomic selection, selecting animals with a reduced genetic risk of twin pregnancies can contribute to reducing the incidence of twin pregnancies in dairy herds.

Sexing of Embryos at the Time of Twin Reduction: A Clinical Approach.

This study provides new unique information on bovine twin pairs during the late embryonic period (28-34 days of pregnancy) in relation to (1) a predictive ultrasound measurement that was differential for sexing heterosexual twins; (2) intrauterine embryonic growth patterns in twin pairs; and (3) a higher vulnerability of female embryos compared to males following an induced embryo reduction in heterosexual twins. The study population comprised 92 dairy cows carrying bilateral twins. A length difference between co-twins equal to or greater than 25% in around 50% of pregnancies served to determine the sex of embryos with 100% accuracy in heterosexual twins, which was assessed four weeks later on the remaining fetus after twin reduction. The apparent rates of growth of twin pairs and of individual male and female embryos from day 28 to 34 of gestation were similar to established growth pattern standards for singletons. Mean embryo sizes in relation to gestational age were smaller by some 5 days' growth equivalent in twins compared to singletons. After the reduction in the female embryo in heterosexual twins, the risk of male embryo loss was null. This new information allowed for sex selection at the time of twin reduction.

Direct transfer of a single frozen-thawed in vitro-produced embryo to avoid twin pregnancy in dairy cows.

Fertility of cow receiving fixed-time artificial insemination (FTAI) was compared with cow in which a single frozen-thawed in vitro-produced (IVP) embryo was fixed-time transferred (FTET) to avoid twin pregnancy. The study population was comprised of 596 lactating dairy cows synchronized for oestrus: 440 were fixed-time inseminated (AI cows), and 156 were given GnRH treatment at the time of embryo transfer (ET cows) 8 days post-oestrus. Of the 596 cows, 235 (39.4%) became pregnant: 175 (39.8%) AI cows and 60 (39.8%) ET cows. Twin pregnancy was recorded in 16% of the AI pregnant cows (28/175), whereas no ET cows had twins (0/60). Significant interaction (p < .01) was observed between breeding technique (FTAI vs FTET) and repeat (RB) or not repeat breeding (NRB) for the likelihood of pregnancy. This meant that using RB AI cows as reference, the odds ratio for pregnancy in RB ET cows was 2.2 (p = .04). In conclusion, transfer of a frozen IVP embryo proved useful to prevent the risk of twin pregnancy without affecting fertility.

Revisiting the Timing of Insemination at Spontaneous Estrus in Dairy Cattle.

Cows show spontaneous estrus over 8-20 h but become refractory to the bull about 10-12 h before ovulation. This indicates that ovulation occurs 10-12 h after the end of estrus behavior, yet spermatozoa from the bull ejaculate need to undergo maturation and capacitation for 6 to 8 h in the female reproductive tract before they are capable of fertilization. Traditionally, the onset of estrus has been considered the best timing for artificial insemination (AI) in cattle, that is, 6 to 24 h from the first signs of estrus. However, recent findings suggest this interval should be reduced to 16 to 6 h before ovulation, bringing it closer to the end of estrus. In this review, the end of estrus rather than its onset is proposed as the best guide for AI timing in dairy cattle, and physiological indicators of late estrus are discussed such as relaxation of the intravaginal part of the uterus, a lower cervical mucus viscosity and a softer pre-ovulatory follicular consistency as simple cues indicating a cow is ready for service.

Twin reduction in the late embryonic period prevents the condition of freemartin in dairy cattle.

The condition of freemartinism occurs in the bovine heterosexual twin foetuses and refers to the resulting infertile female. Vascular anastomoses of the foetal membranes are the major reason of this anomaly. This study examines whether single born heifers following induced twin reduction at 28-34 days of pregnancy could develop normal reproductive functions. The study population derived from 367 lactating dairy cows carrying: unilateral twins (n = 178), bilateral twins (n = 174) or triplets (n = 15), in which manual embryo reduction was performed. The final study population was constituted of 95 single born twin females that reached 12 months of life and entered into the AI period. Of these heifers, 40 have had one unilateral co-twin, 49 one bilateral co-twin, and 6 two contralateral co-twins. A total of 1688 heifers inseminated during the same period were used as controls to compare the rates of pregnancy at 15 months of age, culling due to infertility and birth before the age of 24 months. No differences were detected between groups. With an accepted incidence rate of 50% heterosexuality for all twin sets, we should assume that half of our study population had a male co-twin. Our results indicate lack of effect of the male co-twin on normal development of the gonads and genital tract of his female partner during the late embryonic period.

Clinical Overview of Luteal Deficiency in Dairy Cattle.

Luteal deficiency is defined as reduced progesterone (P4) steroidogenesis by the corpus luteum (CL), either in the amount or duration, or both. This work provides a clinical overview of the current understanding of luteal deficiency and its association with low fertility in dairy cows. Low plasma P4 concentrations during the luteal phase post-artificial insemination (AI) are associated with lower conception rates. Treatments post-AI with P4, gonadotropin-releasing hormone (GnRH) or human chorionic gonadotropin (hCG) improve fertility in some conditions. Sub-luteal function during the late embryonic period (at pregnancy diagnosis, i.e., 28-34 days post-AI), is just one factor among other factors associated with pregnancy loss. Treatment with P4 in cows with one CL favors pregnancy maintenance, while GnRH treatment does the same in cows carrying twins. The diagnosis of sub-luteal function can be made clinically on the basis of plasma or milk P4 concentrations. Automated in-line milk P4 analysis systems to diagnose luteal activity emerge as a very interesting tool in dairy herds. Monitoring plasma or milk P4 concentrations with the help of Doppler ultrasonography to assess the CL function would allow individualizing the luteal phase support.

The Uterus as an Influencing Factor for Late Embryo/Early Fetal Loss-A Clinical Update.

Here we revise circumstances of non-infectious causes in which the uterus may be associated with pregnancy loss during the late embryo/early fetal period (following a positive pregnancy diagnosis in lactating dairy cows). As the uterine size increases with parity and pregnant heifers with no detrimental effects of a previous parturition, a primigravid uterus is proposed as a reference for identifying risk factors that negatively influence pregnancy in lactating cows. Cows suffering placenta retention or with a large uterus at insemination were selected as topics for this revision. Retained placenta, that occurs around parturition, has a long-lasting influence on subsequent pregnancy loss. Although retained placenta is a particularly predisposing factor for uterine infection, farm conditions along with cow factors of non-infectious cause and their interactions have been identified as main factors favoring this disorder. A large uterus (cervix and uterine horns lying outside the pelvic cavity) with no detectable abnormalities has been associated with low fertility and with a greater incidence of pregnancy loss. A large reproductive tract may well derive from an inadequate uterine involution. Therefore, peripartum management and strategies to reduce the incidence of uterine disorders should reduce their associated financial losses in the herds.

Follicular Size Threshold for Ovulation Reassessed. Insights from Multiple Ovulating Dairy Cows.

In Bos. taurus cattle, follicular deviation to dominance begins when the selected ovulatory follicle reaches a mean diameter of 8.5 mm. The dominant follicle acquires the capacity to ovulate when it reaches a diameter of about 10 mm. In this study, data derived from 148 cows in estrus with one follicle of ovulatory size and another of 7-9 mm, reveal that the small follicle has the capacity to ovulate alone or with the dominant follicle; thus, giving rise to a single or twin pregnancy. This indicates that a follicle of deviation size may ovulate in the presence of a follicle of ovulatory size.

Transfer of a single fresh in vitro-produced embryo may prevent twin pregnancy without compromising the fertility of the cow.

This study examines whether the transfer of a fresh in vitro-produced (IVP) embryo can avoid the risk of twin pregnancy without reducing the fertility of a cow. The study population was comprised of 416 lactating dairy cows synchronized for oestrus: 294 were fixed-time inseminated (AI cows), and 122 were given GnRH treatment at the time of embryo transfer (ET) an IVP embryo (ET cows). Of the 416 cows, 167 (40.1%) became pregnant. Twin pregnancy was recorded in 20.8% of the AI pregnant cows (21/101), whereas no ET cows had twins (0/66). Significant interaction (p < .01) was observed between breeding technique and the period of the year for the likelihood of pregnancy. This meant that using AI cows during the warm period (May-September) as reference, the odds ratio for pregnancy in ET cows during the warm period was 3.4 (p = .001). In conclusion, transfer of a single fresh IVP embryo proved useful to prevent the risk of twin pregnancy without affecting fertility.

Ovarian response to prostaglandin F2α in lactating dairy cows: A clinical update.

Prostaglandin F2α (PGF2α) and its analogs are used to induce luteolysis in estrus synchronization programs to terminate unwanted pregnancies or to promote ovulation in certain cow subpopulations. In the past few decades, the luteolytic dose of PGF2α has remained unchanged. This review explores the clinical implications of increasing the standard dose for these applications in high-producing dairy cows. Ultrasonography may assist in selecting the most appropriate PGF2α dose and improve the results. A reference has been used for PGF2α for promoting ovulation in herds showing poor reproductive performance.

Effects of Heat Stress on Follicular Physiology in Dairy Cows.

Follicular organization starts during mid-to-late fetal life with the formation of primordial follicles. The bilateral interplay between the oocyte and adjoining somatic cells during follicular growth and ovulation may be sensitive to heat stress (HS). Mechanisms giving rise to pre-ovulatory temperature gradients across reproductive tissues are mostly regulated by the pre-ovulatory follicle, and because the cooling of the gonads and genital tract depends on a counter-current transfer system of heat, HS may be considered a major factor impairing ovulation, fertilization and early embryo development. There is evidence of a long-lasting influence of HS on oogenesis and final follicular maturation. Follicular stages that are susceptible to HS have not been precisely determined. Therefore, the aim of this review was to describe the influence of HS during the staged follicular development in dairy cattle, from the activation of primordial follicles to ovulation. Some clinical prospects are also considered.

Unilateral twin pregnancy: A non-infectious factor required for the etiological diagnosis of abortion in dairy herds.

Twin pregnancies are classified into bilateral (one fetus in each uterine horn: 44%) and unilateral (both fetuses in the same uterine horn, right or left: 56%). The incidence of abortion during mid- to late gestation is approximately 1% in cows carrying bilateral twins and more than 40% in cows carrying unilateral twins. In this period, abortion seems most commonly associated with infectious agents. However, although this imbalanced abortion rate may imply that unilateral twin pregnancy is a non-infectious abortion factor, few available data can describe the cause of abortions in twin pregnancies. The current findings suggest that unilateral twin pregnancy is a non-infectious factor required for the etiological diagnosis of abortion in dairy herds.

Response to Therapeutic Abortion in Lactating Dairy Cows Carrying Dead Twins during the Late Embryo/Early Fetal Period.

This study examined the dose-response effect of prostaglandin F2α (PGF2α) when used to induce abortion in cows with two dead twins at pregnancy diagnosis 28-34 days post-AI (late embryonic period: LE) or at pregnancy confirmation 49-55 days post-AI (early fetal period: EF). The study population consisted of 415 cows receiving a single PGF2α dose (PG1 group: 254 LE and 161 EF cows) and 200 receiving a 1.5 × PGF2α dose (PG1.5 group: 121 LE and 79 EF cows). The induced abortion rate was significantly lower (chi-square test; p < 0.0001) in the EF (34.6%) than LE cows (88%), and was also reduced (p = 0.001) in the EF PG1 group (28%) compared with the EF PG1.5 group (48.1%). High milk production (≥45 kg) was the only factor influencing the induced abortion rate in LE cows (odds ratio 0.2; p < 0.0001), whereas the odds ratio for induced abortion of PG1.5 cows was 2.3 (p = 0.005) in EF cows. In conclusion, an increased PGF2α dose had no effect on abortion induction during the late embryonic period and its rate was inversely associated with milk production. This higher PGF2α dose did, however, offer benefits during the early fetal period.

Cervix-rectum temperature differential at the time of insemination is correlated with the potential for pregnancy in dairy cows.

This study sought to establish whether temperature gradients between the cervix, vagina, and rectum at and 7 days post-artificial insemination (AI) were associated with the incidence of pregnancy in lactating dairy cows (Experiment I; n = 90 ovulating cows) and to evaluate temperature gradient dynamics from the time of insemination to 7 days post-AI under heat stress conditions (Experiment II; n = 16 ovulating and 4 non-ovulating cows). In Experiment I, 39 cows (43.3%) became pregnant. The odds ratio for pregnancy was 2.5 for each one-tenth of a degree drop in cervical temperature with reference to the control rectal temperature at the time of AI (P = 0.01), whereas the same decrease in the cervix-rectum temperature differential 7 days post-AI resulted in an odds ratio of 0.44 (P = 0.02). In Experiment II, 5 of the ovulating cows (31.3%) became pregnant. The mean values of the vagina-rectum, vagina-cervix, and cervix-rectum temperature differentials at AI (day 0), 8 h, 24 h, and 7 days post-AI changed significantly from day 0 to day 7 (within-subject effect; P < 0.02) in ovulating cows but not in non-ovulating cows. Temperature differentials on days 0 and 7 were similar between ovulating cows and cows of Experiment I. Overall, our findings support the notion that a temperature differential between the caudal cervical canal and rectum at AI may be an indicator of the likelihood of pregnancy. Possible prospects of confirming estrus at the herd-level are also suggested.

Inducing ovulation with human chorionic gonadotrophin improves the pregnancy rate in lactating dairy cows receiving an in vitro-produced embryo.

While the global use of in vitro-produced embryos in dairy cattle is on the rise, several technical aspects of embryo transfer procedures have not yet been optimized. This study compares the effects of inducing ovulation using human chorionic gonadotropin (hCG) versus gonadotropin-releasing hormone (GnRH) at the end of a 5-day progesterone(P4)-based protocol for oestrous synchronization on the pregnancy rate of lactating dairy cow recipients of in vitro-produced embryos. Fresh embryos were transferred on Day-seven post-oestrus to ovulating cows receiving GnRH or hCG (groups GnRH and hCG, n = 60 each). Pregnancy was diagnosed by ultrasound on Day 28 post-oestrus. Forty-nine cows became pregnant: 16 in GnRH (26.7%) and 33 in hCG (55%). Taking GnRH-treated cows as reference, the odds ratio for pregnancy of hCG-treated cows was 3.3 (p = .002). In conclusion, hCG treatment given at the end of a 5-day P4-based protocol for oestrous synchronization improved the pregnancy rate in lactating dairy cows receiving an in vitro-produced embryo.

Presence of multiple corpora lutea affects the luteolytic response to prostaglandin F2α in lactating dairy cows.

Since the 1970s, luteolytic doses used for synchronizing estrus in dairy cattle have remained unchanged. This study aimed to evaluate the dose-response effect of prostaglandin F2α (PGF2α), which is used for synchronizing estrus, and subsequent fertility in cows with two or more corpora lutea (CL). The study population consisted of 1,683 cows with a single CL (1CL), 501 cows with multiple CL receiving a single dose of PGF2α (2CL1), and 252 cows with multiple CL receiving a 1.5 × PGF2α dose (2CL1.5). Cows with a single CL (n = 1,245) showed estrus significantly (P < 0.01) earlier (3.01 ± 1.23 days; mean ± SD) than cows with multiple CL (n = 287; 3.33 ± 1.69 days). Using 1CL cows as reference, the odds ratio (OR) for the estrus response in 2CL1 cows was 0.13 (P < 0.0001), whereas the ORs for estrus response and pregnancy of 2CL1.5 cows were 1.8 (P = 0.0001) and 1.7 (P = 0.001), respectively. Based on the results for only the 2CL1 cows, the OR for the estrus response was 0.7 (P = 0.01) for cows producing ≥ 45 kg of milk at treatment, compared to the remaining cows producing < 45 kg of milk. Our results showed that the presence of multiple CL reduced the estrus response to that induced by a single PGF2α dose and milk production was inversely associated with this response, whereas an increased PGF2α dose improved the estrus response. Therefore, an increase in the standard PGF2α dose is recommended.

Thermal Mechanisms Preventing or Favoring Multiple Ovulations in Dairy Cattle.

While cows are predominantly monovular, over the past 30 years the incidence of multiple ovulations and thus twinning has increased considerably alongside milk production. Multiple pregnancies are not desirable as they negatively affect the health of cows and the herd economy. Although causal mechanisms associated with multiple ovulations have been extensively revised, the process of multiple ovulations is not well understood. Recent studies on the thermal biology of the reproductive system have shown how thermal mechanisms may prevent or favor multiple ovulations. This review focuses on this relationship between thermal dynamics and multiple pregnancies. Cooling of the pre-ovulatory follicle is able to regulate ovulation. In effect, pre-ovulatory local cooling of the female reproductive system favors male and female gamete maturation and promotes fertilization. Thermal stress is proposed here as a model of stress. Periods of high ambient temperature affect the processes of pre-ovulatory follicular cooling and multiple ovulations. While the ratio between unilateral and bilateral multiple pregnancies is normally close to one, under heat stress conditions, this ratio may be 1.4 favoring unilateral multiple pregnancies. A ratio approaching unity is here proposed as an indicator of cow wellbeing.

Benefits and Risks of Preventing Twin Pregnancies.

Clinical problems associated with twin pregnancies have been well established, and twin births are now considered undesirable or even disastrous for the dairy cattle industry and the individual cow. The high incidence of early fetal loss, abortion during the mid-lactation period, dystocia, stillbirth, and placenta retention should be considered a preventable consequence of management, as these disorders greatly compromise the welfare and productive lifespan of a cow carrying or delivering twins. The use of sexed semen generates herd replacements and additional heifers, so a proposed strategy for twin pregnancy prevention is the transfer of a single in vitro-produced female beef cow embryo to cows not suitable for producing replacements. Another proposed strategy is drainage at insemination of co-dominant follicles to prevent twin pregnancies in cows with genetic merit. As a result, embryo survival should improve, economic losses associated with twin pregnancies will be prevented, beef output from the herd will be increased, and the health and welfare of the cow will certainly benefit. In this review, the clinical prospects of preventing or avoiding twin pregnancies using both procedures are discussed.