Association of genomic prediction for twin pregnancies with the incidence of double ovulation in primiparous dairy cows.

Twin pregnancies compromise the health and well-being of dairy cattle. A recent genomic prediction model for twin pregnancies has been developed based on twin calving or abortion. However, the incidence of double ovulation is significantly higher than that of twin births. This study aimed to evaluate whether genomic prediction values for twin pregnancies are associated with the incidence of double ovulation in primiparous dairy cows. Factors influencing the double ovulation rate were analysed using binary logistic regression on 676 cows: 475 (70.3%) inseminated at spontaneous estrus and 201 following one of two different estrus synchronization protocols for fixed-time artificial insemination (FTAI). The odds ratio for double ovulations was 0.92 (p = .002) per unit increase in prediction value and 2 (p = .01) for cows subjected to an FTAI protocol. Our findings suggest that genomic prediction values for twin pregnancies can effectively identify the risk of double ovulation at the herd level.

Cow and management factors and their association with estimates of uterine size and position at the time of insemination of dairy cattle.

This study examines factors affecting uterine size and position determined at 30-36 days postpartum in dairy cattle. The final study population consisted of 328 dairy cows, all calving during the warm season. Uterus position (pelvic, pelvic-abdominal, abdominal) and uterus size (small, medium, large) was measured by ultrasound on Day 30-36 postpartum. Multiparous cows had a larger uterus positioned in the abdominal cavity (p = .03) and a male newborn was associated with a larger uterus (p = .022). The number of cows with the uterus in the abdominal cavity was higher among multiparous and high producer (p < .0001) cows. High producers were 0.41 times more likely to have a small uterus in a pelvic position. The most important finding of this study was that the delivery of a male dairy, rather than beef, calf was related to the least optimal maternal uterus size and position.

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.

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.

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.

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.

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.

Improved embryo survival following follicular drainage of subordinate follicles for twin pregnancy prevention in bi-ovular dairy cows.

To prevent twin pregnancies in cattle, a simple transvaginal device can be used for follicular drainage. This study examines embryo survival following follicular drainage of the smaller pre-ovulatory follicle at timed artificial insemination (AI) in cows with a pre-ovulatory follicle in each ovary. The study groups established were a control group of 289 monovular cows, a non-drainage group of 114 bi-ovular cows and a follicular drainage group of 113 bi-ovular cows. All cows undergoing drainage developed a corpus luteum (CL) in the drained ovary. Pregnancy loss was recorded 56 days post-AI in 19.5% of the 149 cows that became pregnant. Pregnancy loss in the drainage group cows not suffering heat stress (3.8%) was significantly lower (P < 0.05) than in control non-heat stressed cows (20.9%) and heat-stressed non-drainage group cows (25%). Results indicate that CL induction by follicular drainage for twin pregnancy prevention may reduce the incidence of pregnancy loss.

Transfer of a single embryo versus drainage of subordinate follicles to prevent twin pregnancies in dairy cows. Why not both?

In this study, we present two proposed approaches to prevent twin pregnancies in dairy cattle: 1) single, in vitro-produced embryonic transfer into a recipient cow or 2) subordinate follicle drainage at the time of insemination. Both procedures lead to improved embryonic survival. As the use of sexed semen generates herd replacements and additional heifers, we propose the transfer of a single female cattle embryo into cows that are not suitable for producing replacements, and follicular drainage in lactating cows with genetic merit. This should eliminate economic losses associated with twin pregnancies and increase cattle output of the herd.

Effects of twin pregnancy prevention strategies such as GnRH dose and drainage of the smaller follicle on ovulation in dairy cows with two follicles of pre-ovulatory size in the same ovary.

We examined the effects of a single or 2.5-fold dose of dephereline [a gonadotropin-releasing hormone (GnRH) analogue] as well as the drainage of the smaller follicle at the time of insemination on ovulation in dairy cows with two follicles of pre-ovulatory size in the same ovary. The three study groups included 220 monovular cows (control), 110 non-drained cows with two follicles, and 110 cows with two follicles, of which one was drained. In each group, cows treated with a single dose or 2.5-fold dose of dephereline showed similar results following treatment. Ovulation failure of the non-drained follicle occurred in 29.1% of the drained cows, whereas ovulation occurred in 96% of the non-drained and control cows. Twin pregnancy was recorded in 19.4% of the pregnant non-drained cows with two follicles. In conclusion, the increased dephereline dose did not improve the ovulation rate in any group. Follicular drainage, however, prevented twin pregnancy in cows with two follicles, but also resulted in an increase in the non-drained follicle's rate of ovulation failure.

Abortion in dairy cattle with advanced twin pregnancies: Incidence and timing.

This study examines incidences and timing of abortion in Holstein Friesian dairy cows with advanced twin pregnancies in north-eastern Spain. The study population consisted of 1,194 twin pregnancies in two herds recorded from 2010 to 2015: 522 bilateral and 672 unilateral. The presence of live twins was determined by transrectal ultrasonography between 55 and 61 days post-AI and pregnancy confirmed 60 days later. Abortion was recorded in 278 (23.3%) cows before Day 260 of pregnancy: 7 (1.3%) in bilateral and 271 (40.3%) in unilateral twin pregnancies. Using binary logistic regression procedures the following were excluded as risk factors for abortion: herd, year, heat stress (temperature-humidity index values >72), lactation number and right versus left side for unilateral twins. Based on odds ratios, cows carrying unilateral twins were 49.9 (right unilateral twins) and 49.3 (left unilateral twins) times more likely to suffer abortion than cows with bilateral twins. The average time of abortion for unilateral twins was 173 ± 32 days and ranged from 135 to 249 days, 43% of abortions occurring between 135 and 154 days of gestation. The authors suggest prevention or reduction of unilateral twin pregnancies by follicular puncture and drainage of subordinate follicles at AI or manual rupture of the amniotic vesicle of one of the two twins at the time of pregnancy diagnosis.

The effects of a single or double GnRH dose on pregnancy survival in high producing dairy cows carrying singletons or twins.

Pregnancy loss during the late embryonic and early fetal periods influences dairy herd economy. The objectives of this study were to (1) investigate the effects of a single or double GnRH dose administered at the time of pregnancy diagnosis (28-34 days post-AI) on the pregnancy survival of cows in their third lactation or further carrying live singletons or unilateral twins, and (2) examine the impacts of GnRH treatment on subsequent twin reduction in twin pregnancies. Cows carrying singletons (n = 1,054) or unilateral twins (n = 379) were assigned at the time of pregnancy diagnosis to the following groups: control (no treatment), GnRH (100 µg GnRH), and 2GnRH (200 µg GnRH). Pregnancy loss was recorded in 180 of the 1,433 cows (12.6%) at 58-64 days post-AI. Based on the odds ratios, there was a significant (P < 0.0001) interaction between the treatment group and twin pregnancy. This interaction implies that control cows carrying twins were 3.2 times more likely to suffer pregnancy loss than the other cows, whereas the GnRH and 2GnRH treatment groups cows carrying singletons or twins had pregnancy loss rates similar to the control cows carrying singletons. Twin reduction was observed in 35 twin pregnancies (9.2%). Cows in the GnRH and 2 GnRH groups were seven times more likely to show twin reduction than control cows. Our results indicate that GnRH administered at the time of pregnancy diagnosis had no beneficial effects in cows carrying singletons. In contrast, for twin pregnancies, the treatment increased the rate of pregnancy survival and was accompanied by an increase in the twin reduction rate.

Is twin pregnancy, calving and pregnancy loss predictable by serum pregnancy-specific protein b (pspb) concentration 28-35 days after ai in dairy cows?

Double ovulation occurs more frequently in multiparous cows with high milk production than in primiparous cows and the rate of twin pregnancy/calving is increasing worldwide. Diagnosis of twin pregnancy is possible by ultrasound at the time of early pregnancy examination [28-34 days after artificial insemination (AI)]. Pregnancy proteins are also well-known indicators of gestation. The risk of pregnancy loss during the first trimester of gestation for cows carrying twins is three to nine times higher than for cows carrying singletons. Pregnancy-specific protein B (PSPB) is a good indicator not only of pregnancy but also of pregnancy loss. The aims of this study were (a) to collect calving data in some Hungarian Holstein-Friesian herds (n = 7,300) to compare PSPB serum concentrations (measured 29-35 days post insemination) in twin- and singleton-calving cows (Trial 1), and (b) to check the predictive value of PSPB serum concentration for twin pregnancy and pregnancy loss in high-producing Spanish Holstein-Friesian cows (n = 98; Trial 2). Our results showed almost 7% twin calving rate. Although hormonal treatments are commonly believed to be major causes of twin pregnancies, our data do not support this hypothesis. The only exception is the single PGF injection, which significantly increased twin calving. No effect of milk production on the risk of twin pregnancy was found, and twin pregnancy increased with parity. The AI bull, the bull's sire, the bull's grandfather and the cow's father also affected twin calving (P ≤ 0.02). We found much higher frequency of twin calving in cows diagnosed pregnant with higher than 3 ng/ml serum PSPB concentrations at 29-35 days after insemination. In Trial 2, non-significant but well-marked differences were found in PSPB serum concentration between singleton- and twin-pregnant cow samples (2.1 and 2.9 ng/ml) at different bleeding times. Probably the small size of the study population and the effects of milk production on PSPB values may explain this lack of significance.

Foetal death in naive heifers inoculated with Neospora caninum isolate Nc-Spain7 at 110 days of pregnancy.

Neospora caninum infection is a leading cause of abortion in cattle worldwide. The pathogenesis of bovine neosporosis, particularly during the second term of gestation when most abortions occur in naturally infected dams, is poorly understood. In the present study foetal death was observed in 3 of 6 experimentally infected dams at 110 days of gestation after 6 weeks of experimental period. All experimental heifers were febrile between 3 and 5 days post infection (dpi). Inoculated dams seroconverted by 3-4 weeks post-infection with higher mean antibody titres in aborting dams compared to non-aborting heifers, although not significantly (p > 0.05). Neospora caninum DNA was detected in all infected foetuses and placentas, and three infected foetuses also had N. caninum antibodies. The parasite burden was higher in the brain of dead/aborted foetuses than in live foetuses. Interestingly, high IFN-γ production was detected in foetal fluids of a dead foetus found upon euthanasia of its dam, while no IFN-γ was observed in amniotic, allantoic and/or foetal fluids in the three infected foetuses that were alive upon maternal euthanasia. The present study confirms that the infection of dams on gestation day 110 with 10(7) tachyzoites of the Nc-Spain7 isolate causes abortion. The fact that some infected dams aborted and some did not is relevant to the understanding of N. caninum pathogenesis of abortion in naturally infected cows.

Effects of different five-day progesterone-based synchronization protocols on the estrous response and follicular/luteal dynamics in dairy cows.

This study compared the responses shown by lactating dairy cows to four different P4-based protocols for AI at estrus. Cows with no estrous signs 96 h after progesterone intravaginal device (PRID) removal were subjected to fixed-time AI (FTAI), and their data were also included in the study. In Experiment I, follicular/luteal and endometrial dynamics were assessed every 12 h from the beginning of treatment until AI. The estrous response was examined in Experiment II, and fertility was assessed in both experiments. The protocols consisted of a PRID fitted for five days, along with the administration of different combinations of gonadotropin releasing hormone (GnRH), equine chorionic gonadotropin and a single or double dose (24 h apart) of prostaglandin F2α. In Experiment I (40 cows), animals receiving GnRH at the start of treatment showed a significantly higher ovulation rate during the PRID insertion period while estrus was delayed. In Experiment II (351 cows), according to the odds ratios, cows showing luteal activity at the time of treatment were less likely to show estrus than cows with no signs of luteal activity. Treatment affected the estrous response and the interval from PRID removal to estrus but did not affect conception rates 28-34 days post AI. Primiparous cows displayed a better estrous response than multiparous cows. Our findings reveal acceptable results of 5-day P4-based protocols for AI at estrus in high-producing dairy cows. Time from treatment to estrus emerged as a good guide for FTAI after a 5-day P4-based synchronization protocol.

Coxiella burnetii antibody dynamics in heifers born to vaccinated versus non-vaccinated dams in a chronically infected dairy herd.

This study was designed to compare Coxiella burnetii antibody dynamics in heifers born to vaccinated or non-vaccinated dams in a single high-producing dairy herd chronically infected with the bacterium. Antibody dynamics were examined from birth to the postpartum period in replacement heifers (n = 14) born to non-vaccinated dams (n = 7) or to dams that had been vaccinated on gestation days 171-177 (n = 7) and 192-198. Samples of blood, milk, faeces, vaginal fluid, colostrum and cotyledons (the latter two only at parturition) were obtained in the dams over the period from gestation days 171-177 to postpartum days 91-97. Blood samples were used to detect antibodies against C. burnetii and remaining samples for PCR identification of the bacterium. In their calves/heifers, blood samples for antibody determinations were collected from birth to postpartum at the time points 1-7 and 22-28 days and 3, 6 and 12 months of age; 90-96 and 210-216 days of gestation; and 22-28 days postpartum. All calves were born seronegative for C. burnetii. Irrespective of the shedding status of their mothers (7 were C. burnetii shedders), seroconversion occurred after colostrum intake in all calves born to seropositive cows (n = 9) and in two of three vaccinated seronegative dams. Thereafter antibody titres gradually declined and by 6 months of age all calves were seronegative. Seronegativity persisted until their first postpartum period. These findings indicate that cows vaccinated during advanced pregnancy transfer immunity to their calves via the colostrum. Maternal C. burnetii antibodies in calves persisted for three months in calves born both to seronegative vaccinated and seropositive dams.

Effects of long-term vaccination against Coxiella burnetii on the fertility of high-producing dairy cows.

The impact of long-term vaccination against Coxiella burnetii on the fertility of cows was studied. Double vaccinations three weeks apart at the start of the third trimester of gestation in each of two consecutive pregnancies were applied. The final study population consisted of 410 cows after the first vaccination round. Based on the odds ratios, the likelihood of early fetal loss (pregnancy loss following a positive pregnancy diagnosis before Day 90 of gestation) was higher in control cows (OR = 1.42) than in vaccinated cows. The final study population consisted of 336 cows after the second round of vaccination. According to the odds ratios, vaccinated C. burnetii seronegative cows were less likely to be subfertile (> 3 AI) (OR = 0.4) compared to non-vaccinated seronegative animals, and the likelihood of early fetal loss was lower in vaccinated C. burnetii seropositive animals (OR = 0.3) compared to non-vaccinated seronegative cows. Seropositivity to C. burnetii was positively related to twin pregnancy after the two rounds of vaccination (OR = 2.1 and 3.5, respectively). These results indicate that two consecutive vaccination rounds against C. burnetii in advanced gestation reduce subfertility and early fetal loss in dairy cows.

Effects of different five-day progesterone-based fixed-time AI protocols on follicular/luteal dynamics and fertility in dairy cows.

This study compares in two experiments the responses of lactating dairy cows to four different progesterone-based protocols for fixed-time artificial insemination (FTAI) in terms of their effects on follicular/luteal dynamics and fertility. The protocols consisted of a progesterone intravaginal device fitted for five days, along with the administration of different combinations of gonadotropin releasing hormone, equine chorionic gonadotropin and a single or double dose (24 h apart) of prostaglandin F2α. In Experiment I, the data were derived from 232 lactating cows. Binary logistic regression identified no effects of treatment on ovulation failure or multiple ovulation 10 days post artificial insemination (AI). Based on the odds ratio, the likelihood of ovulation failure was lower (by a factor of 0.1) in cows showing at least one corpus luteum (CL) upon treatment than in cows lacking a CL; repeat breeders (> 3 AI) and cows with multiple CLs at treatment showed lower (by a factor of 0.44) and higher (by a factor of 9.0) risks of multiple ovulation, respectively, than the remaining animals. In Experiment II, the data were derived from 5173 AIs. The independent variable treatment failed to affect the conception rate 28-34 days post AI, twin pregnancy or early fetal loss 58-64 days post AI. The results of this study demonstrate the efficacy of 5-day progesterone-based protocols for FTAI. All four protocols examined were able to induce ovulation in both cyclic and non-cyclic animals so that FTAI returned a similar pregnancy rate to spontaneous estrus. Our results suggest that the ovarian response and fertility resulting from each treatment are due more to the effect of ovarian structures at treatment than to the different combinations of hormones investigated.

Serological and shedding patterns after Coxiella burnetii vaccination in the third gestation trimester in dairy cows.

This study sought to assess the effects of an inactivated phase I vaccine against Coxiella burnetii at the start of the third trimester of gestation on serological profiles, bacterial shedding patterns and subsequent reproductive performance in dairy cows. Cows were randomly assigned to a control (n = 78) or a vaccinated (n = 78) group on days 171-177 of gestation. Samples of placenta and colostrums at parturition, vaginal fluid, faeces, milk (PCR identification) and blood (anti-C. burnetii antibody detection) were obtained on the day of treatment and on days 91-97 post partum, and also on parturition day and weekly on days 1-7, 8-14, 15-21, 22-28 and 29-35 post partum in a subset of 70 animals. By Kaplan-Meier survival analysis, no significant effect of vaccination was detected on any of the reproductive variables studied. According to the odds ratio, C. burnetii shedding on days 171-177 of gestation was highly correlated with seropositivity against C. burnetii (OR = 9.1), while vaccination was not linked to reduced shedding of the bacterium. In shedders compared to others, the likelihood of pregnancy to first AI decreased and increased by factors of 0.26 and 16.1 on days 1-35 and 91-97 post partum, respectively. In conclusion, when administered at the start of the third trimester of pregnancy, the inactivated C. burnetii phase I vaccine failed to reduce bacterial shedding.