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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.

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.

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.

The effects of season on the fertility of the dairy cow added to the metabolic stress of milk production are well known. We here present lactating dairy cows as a comparative model of this problem. This review examines the results of recent studies that have highlighted heat stress (HS) effects on pre-ovulatory follicles. From these studies, we draw information regarding the mechanisms giving rise to temperature gradients across reproductive tissues. Our review is completed by a discussion of approaches designed to reduce the negative effects of HS based on cooling strategies implemented before ovulation at or around estrus.

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.

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.

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.

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.

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.

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 &lt, 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, 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.

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.

Over the past few decades, the luteolytic dose of prostaglandin F2α (PGF2α) and its analogs, used to synchronize estrus for fixed-time insemination in dairy cattle, have remained unchanged. Given the beneficial effects of PGF2α on a young corpus luteum and on multiple ovulations in a fixed-time insemination protocol, and its therapeutic abortive effects on multiple ovulations in pregnant cows, we propose the use of a double PGF2α dose or two PGF2α treatments 24 hours apart. Ultrasonography procedures serve to identify luteal structures and may therefore help to determine the best PGF2α dose to improve the fertility of high-producing 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. This study received financial support from the procedure “01.02.01 de Transferència Tecnològica del Programa de desenvolupament rural de Catalunya 2014-2020” (Number 19005).

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.

L.E.A.Rowson (1914–1989) raised horizons in farm animal research and the impact of his reproductive contributions remains of major importance worldwide. First, his studies with E.J.C. Polge on the successful deep – freezing and thawing of bull spermatozoa enabled artificial insemination to be applied on an international scale. Second, his development of non-surgical embryo transfer techniques in cattle gave a major boost to the breeding industry, facilitated by deep -frozen storage of embryos during international transport. He also guided many research students whose work in cattle, sheep and horses has become prominent during the past 60 years.

Simple Summary While cows are usually monovular, the incidence of dizygous twin births has recently increased considerably alongside increasing milk production. Genetic progress and improvements in nutrition and management practices have led to a continuous increase in milk yield and thus also to multiple ovulations and twin pregnancies. Twin pregnancies are undesirable as they seriously compromise the welfare of the cow and herd economy. A better understanding of the processes associated with multiple ovulations should help to reduce rates of twinning. During the stages of the sexual cycle, temperature gradients are established within the ovary and throughout the genital tract. Pre-ovulatory local cooling of the reproductive system favors male and female gamete maturation and subsequent fertilization. In fact, thermal mechanisms may prevent or favor multiple ovulations and thus twinning. The purpose of this review was to update this topic. Abstract 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.

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.

Simple Summary Environmentally induced hyperthermia, also called heat stress (HS), compromises reproductive physiology in mammals. The number of oocytes is fixed after birth and they are stored in the ovary in a quiescent state (at the stage of the first meiotic prophase) in primordial follicles. There is evidence that HS alters the oocyte quality, the dynamics of follicular growth and ovulation. The dairy cow, submitted to the metabolic stress of high milk production, is a good model for studying the effects of HS on ovarian function. The aim of this review is to describe the influence of HS during the stages of follicular development in dairy cattle, from the activation of primordial follicles to ovulation. Some clinical aspects are also considered. Abstract 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.

Simple Summary In dairy herds, twin or other multiple pregnancies are not desirable as they compromise the health and productive lifespan of the cows. The mean productive lifespan of primiparous and secundiparous dairy cows delivering twins is about 300 and 200 days shorter, respectively, than that of cows delivering singletons. In addition, the long-term negative effects of twinning are not limited to the early lactation but continue for a period of up to 800 days after calving. Herd management options after diagnosing a twin pregnancy are discussed from an animal health and economic perspective. Abstract Multiple ovulations and so multiple pregnancies have increased recently in dairy cattle. The incidence of the double ovulation impact in high producers at insemination may be over 20%. Twin pregnancies are undesirable as they seriously compromise the welfare and productive lifespan of the cow and herd economy. Clinical problems extend from the time of pregnancy diagnosis to pregnancy loss, abortion or parturition. Early pregnancy loss or abortion of multiple pregnancies lead in most cases to culling. In cows reaching their term, mean productive lifespan is up to about 300 days shorter for cows delivering twins than for cows delivering singletons. While there is an urgent need to address multiple pregnancy prevention procedures in the foreseeable future, the incidence of twin pregnancies continues to rise in parallel with increased milk production. Herein, we review two contrasting measures proposed for the time of twin pregnancy diagnosis: (1) gonadotropin-releasing hormone treatment for pregnancy maintenance, or (2) embryo reduction. These options are discussed in terms of their implications for individual animal health and herd economy. Our main conclusions find that manual twin reduction has proven to be the best management option, whereas the use of prostaglandin F2α for inducing abortion may be a better option than doing nothing.

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. This study received financial support from the project ‘01.02.01 de Transferència Tecnològica del Programa de desenvolupament rural de Catalunya 2014-2020’ (Number 19005).

Broad biological aspects and accepted evolutionary sequences may offer useful guidance towards a comprehensive explanation of the function of mammalian ovaries and oviducts and their vital contribution to the events of fertilization. Cooling of the preovulatory follicles before ovulation may well have its roots in the primitive stages of external fertilization in which aggregates of oocytes are shed into freshwater.

This study sought to establish whether treatment with the GnRH analog dephereline in the early luteal phase could be useful to improve fertility in repeat-breeder cows (>3 inseminations). Dephereline was given 5–7 days after insemination, and effects of both a single and elevated (2.5 times) dephereline dose on pregnancy and subsequent embryo survival determined in repeat-breeder lactating dairy cows. The study population comprised 810 cows: 399 repeat-breeder cows and 411 non repeat breeders. To determine the effects of treatment on the pregnancy rate, three study groups were established: Control, no treatment (n = 269), DEPH, single dephereline dose (100 μg) (n = 270), or 2.5DEPH, 2.5 dephereline doses (250 μg) (n = 271). Recorded pregnancy rates in these groups were 28.6% (77/269), 31.9% (86/270) and 39.1% (106/271), respectively. Binary logistic regression analysis revealed a significant effect of the interaction treatment x repeat breeder on the pregnancy rate. This meant that the single dose and control treatments led to a pregnancy likelihood of repeat-breeder cows reduced by factors of 0.47 and 0.34, respectively, when compared to the 2.5 dose treatment as reference. Treatment had no effects on non repeat breeders. Based on odds ratios, treatment given either as a single or 2.5 dose increased the probability of an additional corpus luteum (more corpora lutea than embryos) in pregnant cows by factors of 5 or 5.4, respectively. Treatment had no effect on embryo survival. Our findings support the hypothesis that dephereline treatment in the early luteal phase after AI improves fertility in repeat-breeder cows. A dephereline dose 2.5 times higher than normal led to improved pregnancy rates, thus overcoming the impacts of repeat-breeder syndrome. Importantly, both the single and 2.5 dephereline doses increased the chances of pregnant cows having an additional corpus luteum, likely favoring embryo survival.

This study compares the effects of inducing ovulation using human chorionic gonadotropin (hCG) or gonadotropin releasing hormone (GnRH) at the end of a 5-day progesterone(P4)-based protocol for fixed-time artificial insemination (FTAI) in lactating dairy cows on ovarian dynamics and fertility. A P4 intravaginal device (PRID) was fitted for five days and GnRH administered upon PRID insertion and a double dose (24 h apart) of prostaglandin F2α upon PRID removal. Cows received either GnRH (GnRH group; n = 98), 1000 IU hCG (hCG-1 group; n = 97), or 3000 IU hCG (hCG-2 group; n = 104) 36 h after PRID removal and were inseminated 50–56 h after PRID removal. Based on Tukey-Kramer tests, cows in hCG-1 and hCG-2 showed a larger follicle diameter at AI than cows in GnRH (P

While beef cattle farmers welcome twin pregnancies, in the dairy cattle industry twinning is considered undesirable or even disastrous. Clinical problems associated with twin pregnancies have been well reviewed in recent years and certainly affect high milk producing herds. In the present essay, we have tried to highlight a number of interventions directed towards avoiding or preventing twin pregnancies. Veterinarians and skilled technicians are already familiar with some of the individual techniques mentioned, but the time has come for us to consider their applications in a broader context. The valuable approaches to preventing twins discussed here are (1) puncture and drainage of subordinate follicles at the time of insemination, and (2) the use of an embryo transfer technique to implant a single blastocyst of demonstrable viability. Both these approaches need to be accompanied by a realistic assessment of the delicate balance between the support to the embryo an induced additional corpus luteum will offer and the risk this induction will carry of generating a twin pregnancy and its consequent problems. Most importantly, both follicular puncture and the transfer of embryos produced in vitro have been shown to significantly reduce pregnancy loss.

The objective of this study was to determine whether fluid drainage from the subordinate follicles by ovum pickup procedures prevents the risk of twin pregnancy without reducing the fertility of the cow. Lactating dairy cows with at least two follicles over 12 mm diameter located one on each ovary and selected from synchronized groups for fixed-time insemination were assigned to a Control (n = 49) or Drainage (n = 49) group. The largest follicle was considered as the dominant follicle. Draining of all subordinate follicles (≥10 mm) was performed in the Drainage group. All drained follicles developed as a corpus luteum 7 days later. In the Control cows, the presence of two corpora lutea (55.1%) was lower (p = .003) than those in the Drainage cows (87.8%), whereas the incidence of twin pregnancies was 50% and 0% for the Control and Drainage groups, respectively (p < .001). Draining did not affect the pregnancy rate. These results indicate that puncture and drainage of the subordinate follicles at insemination may eliminate the risk of twin pregnancies and reduce the risk of subsequent pregnancy loss by increasing the incidence of additional corpora lutea.

Contents Based on experimental studies and extensive field-scale experience, this review offers new proposals for: (i) elevating the success of modern insemination procedures, (ii) emphasizing features of a pre-ovulatory follicle in the context of optimum insemination timing, (iii) overcoming heat stress and its consequences using physiological processes or endocrine protocols, (iv) establishing a viable pregnancy or early pregnancy loss and (v) the challenge of twin pregnancies. In conclusion, the fertility of high-yielding dairy cows can be further improved.

This study examines the possible effects on the reproductive performance of high-producing dairy cows of progesterone (P4) given in the early luteal phase (1.55 g of P4), from Days 3 to 5 post-artificial insemination (AI) as compared with the time of pregnancy recognition, from Days 15 to 17 post-AI. Cows in their third day post-AI were alternately assigned on a weekly rotational basis to the following groups: control, no treatment (C: n = 351), P4 treatment started 15 days after AI (P4-D15: n = 261), or P4 treatment started 3 days after AI (P4-D3: n = 203). Based on odds ratios, cows in P4-D3 were 1.71 times more likely to conceive than control cows (P = 0.004), whereas cows in P4-D15 showed a 1.4-fold greater risk approaching significance of becoming pregnant compared with control cows (P = 0.06). Differences were not observed between treatments. In nonpregnant cows, the given treatment (D3 vs. D15) had no effect on subsequent return to estrus or AI interval and neither were any effects of treatment observed on early fetal loss rates. In contrast, in pregnant cows, the relative risk of twin pregnancy was 2.5 times higher for those in P4-D15 (P = 0.02) than the remaining cows. These findings indicate the efficacy of P4 supplementation after AI. However, when given at the time of pregnancy recognition rather than in the early luteal phase, this treatment increases the twin pregnancy rate.

In the Northern Hemisphere, from June to September and in the Southern Hemisphere from December to March, there are periods of reduced fertility (sub-fertility) in dairy cows that are described as summer infertility. Several factors contribute to sub-fertility during this time, such as ambient temperature, humidity and photoperiod. During the warm season there is a reduction in feed intake that may compromise the energy balance of the cow and/or induce an imbalance in the activity of the hypothalamo-hypophyseal-ovarian axis. These factors reduce the reproductive performance of the cow and compromise the quality of oocytes, embryos and corpora lutea. This paper reviews current knowledge on the metabolic and endocrine mechanisms that induce summer infertility and describe their effects on follicle, oocyte and embryo development in dairy cows.

Carrying twins has been extensively described as an emerging principal non-infectious factor jeopardizing pregnancy maintenance and reducing the lifespan of dairy cows. The risk of pregnancy loss during the first trimester of gestation for cows carrying twins may be from three to seven times higher than for cows carrying singletons. Longer calving to conception intervals, higher culling rates and a shorter mean production lifespan of 200 days have been reported for cows delivering twins compared to cows delivering singletons. There is therefore a need accurately to detect twin embryos at the time of pregnancy diagnosis to follow twin pregnancies carefully or, alternatively, to remove one of the embryos. Therapeutic approaches for the problem of twin pregnancies include GnRH treatment or induced embryo reduction. With similar results, manual rupture of the amniotic vesicle or transvaginal ultrasound-guided aspiration of allanto-amniotic fluid have been proposed as methods of choice to perform twin reduction in cows on Day 28–41 of gestation. However, benefits and risks of induced twin reduction should be quantified. This report reviews various aspects concerning control of twin pregnancies and the practical implications at herd level. Special attention is paid to timing of spontaneous twin reduction. Prospects for induced embryo reduction are also discussed.

Carrying twins is the main factor jeopardizing pregnancy maintenance and reducing the lifespan of dairy cows. There is therefore a need to accurately detect twin embryos at the time of pregnancy diagfiosis so that twin pregnancies are carefully followed. This study compares in single and twin pregnancies from day 28-42 of gestation, patterns of plasma pregnancy-associated glycoproteins (PAGs) I and II and progesterone (P4), and establishes cut-offs for these hormones on day 28 post-AI that Might be useful for twin pregnancy diagnosis. The sensitivity and specificity of the diagnostic capacity of these cut-off values were then determined using ultrasonography findings as the reference standard. A total of 110 lactating dairy cows carrying live singletons or twins from day 28-42 of gestation were enrolled. On day 28 post AI, 56 cows were diagnosed as carrying twins and 54 singletons by ultrasonography. This diagnosis was confirmed ultrasonographically on days 35 and 42 post AI. Among the 54 cows with single pregnancies, those in which the single embryo was accompanied by two corpora lutea (n=23) maintained this condition until day 42 and showed similar plasma PAG-I, PAG-II and P4 levels as those cows with a single embryo and single corpus luteum (n=31). Cows bearing twins showed higher plasma PAG-I, PAG-II and P4 concentrations throughout the study period than cows bearing singletons (between subject effect; P < 0.001). Cut-offs for plasma PAG-I, PAG-II and P4 concentrations on day 28 of gestation of 5.5, 0.25 and 14 ng/ml, respectively, were used to determine twin pregnancies. Each of the proposed cut-off values identified twin pregnancies with a precision higher than 70%. In conclusion, the determination of these hormones could be a useful tool to discriminate between single and twin pregnancies.

Contents The objective of this study was to determine whether induced luteolysis of one of the two corpora lutea in twin pregnancies would provoke spontaneous twin reduction. In Experiment 1, 12 post-partum cows with two corpora lutea in the same ovary were assigned to (three cows per group): Group I, Group II, Group III or Group IV receiving into one of the corpora lutea puncture with no treatment, 0.5 mg dinoprost, 1.5 mg dinoprost and 2.5 mg dinoprost, respectively. One of the two corpora lutea showed clear signs of luteolysis on Day 2 and was practically non-detectable on Day 7 after treatment in the three cows of the Group IV. In Experiment 2, 11 cows carrying live twins with two corpora lutea on Day 28 of gestation, eight bilateral and three unilateral, received 2.5 mg dinoprost into one of the corpora lutea. Corpus luteum reduction and embryo reduction after treatment were registered in 10 and 9 cows, respectively. In bilateral twin pregnancies, four cows suffering embryo reduction remained pregnant. In unilateral twin pregnancies, membrane detachment resulted in the death of both cotwins. In conclusion, although observations were based on few animals, there seems to be a mechanism that operates locally to transfer ovarian progesterone to the uterus, and also a quantitative relationship between the amount of progesterone secreted and support of conceptuses, resulting in death of one twin embryonic vesicle when one corpus luteum regresses.

This study examines the possible effects of progesterone (P4) supplementation during the time of pregnancy recognition, from Days 15 to 17 post-artificial insemination (AI), on reproductive performance in high-producing dairy cows. Cows in their 15th day post-AI were alternately assigned to a control, no-treatment group (C: n = 257) or treatment group (P4: n = 287) on a weekly rotational basis according to the chronologic order of their gynecologic visit. On the basis of the odds ratio, the interaction treatment × previous placenta retention had a significant effect (P = 0.02) on conception rate. Thus, cows in P4 that had not suffered a retained placenta were 1.6 times more likely to conceive 28 to 34 days post-AI than the remaining cows. In nonpregnant cows, treatment had no effect on subsequent return to estrus or AI interval and neither were any effects of treatment observed on twin pregnancy and early fetal loss rates. The results of this study demonstrate the efficacy of P4 supplementations during the time of pregnancy recognition after AI in cows without a clinical history of placenta retention.

This study was designed to examine (i) the seroprevalence of Mycobacterium avium subs paratuberculosis (MAP) in a high-producing dairy herd with clinical symptoms of bovine paratuberculosis, (ii) MAP seroconversion and seronegativation dynamics in the herd and (iii) possible relationships between MAP infection status and herd reproductive performance. One single blood test per cow was performed early post-partum on a monthly basis from day 10-40 post-partum during the first year of the study in 519 cows belonging to a commercial dairy herd. A subset of 111 cows that became pregnant during the study was tested again 60-200 days later during the early foetal period, immediately after the first confirmation of gestation at 58-64 days post-AI. Logistic regression analysis indicated no effect of any independent variable on MAP seropositivity and conception rate 28-34 days post-AI. MAP seropositivity was not a factor affecting the anoestrous, subfertility and early foetal loss rates. In the subset of 111 cows, animals that seroconverted had a 3.9 times greater risk of suffering from early foetal loss (30.3%, 10/33) than the remaining pregnant animals (10.3%, 8/78), (95% confidence interval: 1.11-13.4; p = 0.003). In conclusion, early foetal loss was positively correlated with seroconversion to MAP. Reproductive performance was not impaired by MAP infection.

Multiple pregnancies have devastating consequences on the herd economy of dairy cattle. This observational study examines incidence patterns based on data from the ultrasonographic examination of 1130 multiple pregnancies in cows in their third lactation or more carrying twins (98.8%), triplets (1.1%), or quadruplets (0.08%), and 3160 of their peers carrying singletons. Cows became pregnant following a spontaneous estrus with no previous hormone treatments. Irrespective of a significant decrease (p &lt, 0.0001) in the conception rate (28&ndash, 34 days post-insemination) during the warm period of the year, the multiple pregnancy rate was similar for both warm (26.5%) and cool (26.3%) periods. The incidence of unilateral multiple pregnancies (all embryos in the same uterine horn) was higher than that of bilateral pregnancies (at least one embryo in each uterine horn): 54.4% versus 45.6% (p &lt, 0.0001). This difference rose to 17% during the warm season (p = 0.03). Pregnancy was monitored in unilateral multiple pregnancies until abortion or parturition (n = 615). In the warm period, the parturition rate was 43% compared to 61% recorded in the cool period (p &lt, 0.0001). Thus, a warm climate is the main factor compromising the fate of multiple pregnancies. Some clinical suggestions are provided.

This study compares the fertility effects of inducing ovulation using the GnRH analogue dephereline versus natural GnRH at the end of a 5-day progesterone(P4)-based protocol for fixed-time artificial insemination (FTAI) in heat-stressed and non-heat stressed anestrous lactating dairy cows. Cows were given GnRH (GnRH group, n = 676) or dephereline (DEPH group, n = 655) and inseminated 12 hours later. Based on odds ratios, pregnancy loss was 3.6 times more likely in cows carrying twins (P

This study compares the fertility 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 fixed-time artificial insemination (FTAI) in heat-stressed and non-heat stressed anestrous lactating dairy cows. Heat stress (HS) was defined as an environmental temperature-humidity index (THI) > 72. A P4 intravaginal device (CIDR) was fitted for five days and GnRH administered upon CIDR insertion and a double dose (24 h apart) of prostaglandin F2α upon CIDR removal. Cows then received either GnRH (GnRH group; n = 506), or hCG (hCG group; n = 493) 36 h after CIDR removal and were inseminated 50-56 h after CIDR removal. Ovulation failure was investigated in a subset of 425 cows: 223 and 202 receiving GnRH and hCG, respectively. Based on odds ratios, the interaction between treatment and HS had a significant effect on the ovulation failure rate (P = 0.01). This meant that compared to the rates recorded in non-heat-stressed, GnRH-treated cows (13%), ovulation failure in heat-stressed GnRH-treated cows (25.3%) was more likely by a factor of 2.3 (P = 0.04), in non-heat-stressed hCG-treated cows (2.3%) was less likely by a factor of 0.16 (P = 0.02) and was not significantly different in heat-stressed hCG-treated cows (7%). Interactions between treatment and HS and between treatment and parity had a significant effect on the pregnancy rate (P = 0.0001 and P = 0.001, respectively). The treatment-HS interaction determined that compared to the rates recorded in non-heat-stressed, GnRH-treated cows (30.5%), pregnancy in heat-stressed GnRH-treated cows (17.6%) was less likely by a factor of 0.48 than the remaining cows (P = 0.001), whereas because of the treatment-parity interaction, compared to the rates recorded in primiparous, GnRH-treated cows (31.4%), pregnancy in GnRH-treated multiparous cows (18.9%) was less likely to conceive by a factor of 0.51 than the remaining cows (P = 0.002). No significant effects of treatment on the rates of pregnancy loss or twin pregnancy were identified by binary logistic regression. In conclusion, hCG treatment given at the end of a 5-day P4-based protocol for FTAI improved ovulation and pregnancy rates in anestrous cows under conditions of HS and also had a beneficial impact on the pregnancy rate in anestrous multiparous cows throughout the year.

This study sought to examine the impact of the presence of two co-dominant (ovulatory) follicles at the time of artificial insemination (AI) on the ovulatory response to GnRH given in a fixed-time AI protocol. The study population comprised 622 lactating dairy cows: 306 (49.2%) with a single follicle, 198 (31.8%) with two bilateral follicles (one follicle per ovary) and 118 (19%) with two unilateral follicles (same ovary). Based on odds ratios, cows with two bilateral or unilateral follicles were less likely (by factors of 0.09 and 0.11, respectively) to undergo ovulation failure compared with cows with one follicle (P = 0.01 and P = 0.02, respectively); the likelihood of ovulation failure decreased 0.75 times with every one-mm increase in follicle diameter for cows with a single follicle, whereas individual follicle diameter was not related to ovulation failure in cows with two bilateral follicles (P = 0.001). The likelihood of double ovulation decreased 0.7 times with every one-mm diameter difference between the larger and smaller follicle for all cows with two follicles (P = 0.001), whereas cows with two unilateral follicles showed a higher (P

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.

Heat stress has consequences on both the physiology and reproductive performance of cows, but the most dramatic effect for dairy producers is the decrease produced in fertility. The effects of heat stress on fertility include an increased number of days open, reduced conception rate, and larger number of cows suffering different types of anestrus. Once becomes pregnant, heat stress affects also the reproductive success of the cow through its direct effects on the ovary, uterus, gametes, embryo, and early fetus. This article reviews current knowledge of the effects of heat stress on fertility in dairy cows and the hormonal strategies used to mitigate these effects at the farm level. Administration of GnRH at the moment of artificial insemination can improve the conception rate. Breeding synchronization protocols for fixed-time insemination may reduce the calving conception interval and the number of services per conception. Progesterone-based protocols seem resolve better the reproductive disorders related to a hot environment (anestrus) than GnRH-based protocols. The use of combinations of GnRH, eCG, and hCG in progesterone-based protocols can improve results. Progesterone supplementation during the late embryonic and/or early fetal period would be useful in curtailing pregnancy losses, mainly in single pregnancies, whereas a more positive effect of treatment with GnRH than progesterone has been found in twin pregnancies. Melatonin therapy is emerging as a promising strategy to improve the natural reproductive performance of cows suffering conditions of heat stress.

Contents Q fever is a zoonosis produced by Coxiella burnetii, a bacterium that is widely distributed worldwide. Domestic ruminants are the most important source of C. burnetii for human infection. In sheep and goats, abortion is the main clinical consequence of infection, yet the symptoms described in cattle have so far been inconsistent. Q fever has been also scarcely reported in cattle, most likely because of its difficult diagnosis at the farm level and because of the many existing responsible C. burnetii strains. In this report, the effects of C. burnetii infection or Q fever disease on the reproductive behaviour of dairy cattle are reviewed, with special emphasis placed on the scarcity of data available and possible control actions discussed.

Equine chorionic gonadotropin (eCG) is a member of the glycoprotein family of hormones along with LH, FSH and thyroid-stimulating hormone. In non-equid species, eCG shows high LH- and FSH-like activities and has a high affinity for both FSH and LH receptors in the ovaries. On the granulosa and thecal cells of the follicle, eCG has long-lasting LH- and FSH-like effects that stimulate oestradiol and progesterone secretion. Thus, eCG administration in dairy cattle results in fewer atretic follicles, the recruitment of more small follicles showing an elevated growth rate, the sustained growth of medium and large follicles and improved development of the dominant and pre-ovulatory follicle. In consequence, the quality of the ensuing CL is improved, and thereby progesterone secretion increased. Based on these characteristics, eCG treatment is utilized in veterinary medicine to control the reproductive activity of the cow by i) improving reproductive performance during early post-partum stages; ii) increasing ovulation and pregnancy rates in non-cyclic cows; iii) improving the conception rate in cows showing delayed ovulation; and finally, iv) eCG is currently included in protocols for fixed-time artificial insemination since after inducing the synchrony of ovulation using a progesterone-releasing device, eCG has beneficial effects on embryo development and survival. The above effects are not always observed in cyclic animals, but they are evident in animals in which LH secretion and ovarian activity are reduced or compromised, for instance, during the early post-partum period, under seasonal heat stress, in anoestrus animals or in animals with a low body condition score.

This study compared the efficiency of a five-day or standard (nine-day) progesterone-based regimen combined with equine chorionic gonadotrophin (eCG) in a fixed-time AI (FTAI) protocol for dairy cows. The data examined were derived from 3577 inseminations conducted in three dairy herds. Animals with no estrus signs detected over 21 days were randomly assigned to a PRID-9 or PRID-5 group. Cows in each group received a progesterone intravaginal device (PRID) for 9 or 5 days, respectively, PGF2α and eCG on PRID removal, and GnRH 48 h later. Fixed-time AI was performed 12 h after the GnRH dose. Cows artificially inseminated following spontaneous estrus during the study period were considered as controls. Based on the odds ratio, the likelihoods of animals in PRID-9 in the warm (conception rate [CR] of 22.3%) and cool (32% CR) periods, and control animals in the warm period (26.6% CR) becoming pregnant were reduced (by factors of 0.6, 0.3 and 0.4, respectively) compared with the control animals in the cool period (CR of 43.7%). The risk of a twin pregnancy was higher (51.4%) for cystic PRID-9 cows (by a factor of 3.6) and lower (9.9%) for cyclic PRID-5 animals (by a factor of 0.4) compared with the PRID-9 cyclic cows. These findings indicate that the proposed protocol achieves similar results during the cool or warm season to those obtained when AI is conducted at spontaneous estrus during the cool season. In addition, PRID-5 reduced twin pregnancy compared with PRID-9.

Heat stress is especially harmful for bovine ovarian follicle development and oocyte competence. In this study, we assessed the effects of heat shock on oocyte maturation in oocytes collected during the cold (February–March; n = 114) or warm (May–June; n = 116) periods of the year. In both cases, cumulus-oocyte complexes were matured under control (38 °C) and heat shock conditions (41.5 °C, 18–21 h of maturation). For each oocyte, nuclear stage, cortical granule distribution and steroidogenic activity of cumulus cells were evaluated. Based on the odds ratio, heat-shocked oocytes were 26.83 times more likely to show an anomalous metaphase II morphology. When matured under heat shock conditions, oocytes obtained in both seasons were similarly affected in terms of nuclear maturation, whereas a seasonal effect was observed on cytoplasmic maturation. For oocytes collected during the cold season, the likelihood to show an anomalous maturation was 25.96 times higher when exposed to the heat treatment than when matured under control conditions. By contrast, oocytes collected during the warm season matured under control or heat shock did not show significant risk of showing an anomalous cytoplasmic maturation. Our findings indicate an increased rate of premature oocytes in response to heat shock as well as a higher tolerance to this stress of oocytes harvested in the warm season compared with those collected in the colder period.

Temperature gradients in female reproductive tissues seem to influence the success of key processes such as ovulation and fertilization. The objective of this study was to investigate whether pre-ovulatory follicles are cooler than neighbouring uterine tissue and deep rectal temperatures in lactating dairy cows under heat stress conditions. Temperatures within the pre-ovulatory follicle, on the uterine adjacent surface and 20 cm deep within rectum, were measured using fine thermistor probes within 45 min after sunrise (dawn). Cows were selected from synchronized groups for fixed-time insemination during the warm period of the year. Five cows under direct sun radiation and 11 cows in the shade were included in the study. None of the cows in the sun area ovulated within 24 hr, whereas 10 of the 11 cows in the sun area ovulated. Four of the 10 ovulating cows became pregnant. In the ovulating cows, follicular temperatures were 0.74 and 1.54°C significantly cooler than uterine surface and rectal temperatures, respectively, whereas temperatures in the uterine area were 0.80°C significantly cooler than rectal temperatures. No significant differences among temperatures were found in non-ovulating cows. Follicular size was similar for ovulating and non-ovulating cows. Environmental temperatures in the shade area were 6.4°C significantly lower than those in the sun area. Results of this study indicate that pre-ovulatory follicles are cooler than neighbouring uterine tissue and deep rectal temperatures and those temperature gradients were not found in cows suffering ovulation failure.

CONTENTS The aim of this study was to determine the effect of melatonin treatment during the early dry-off period on subsequent reproductive performance and milk production in high-producing dairy cows under heat stress conditions. In experiment I, addressing the pharmacokinetics of melatonin treatment in lactating dairy cows, doses of untreated, 3, 6, 9 or 12 implants/animal (18-mg melatonin each implant) were given as subcutaneous implants on gestation day 120-20 multiparous lactating dairy cows (four cows/dose group). Experiment II was performed during the warm season on 25 heifers and 114 high milk-producing Holstein-Friesian cows. Animals were randomly assigned to a control (C) or melatonin group (M). Animals in the M group received nine implants (heifers) or 12 (cows) of melatonin on day 220 of gestation. In experiment I, cows in the 12 implants group showed a higher maximum melatonin concentration (Cmax ) and area under the concentration curve from treatment day 0 to day 49 (AUC0-49d ) than those in the remaining groups, among which there were no significant differences in this variable. In experiment II, the likelihood of repeat breeding syndrome (

Contents The objective of this study was to assess the effects of Coxiella burnetii shedding or seropositivity on post-partum recovery and subsequent fertility in high-producing dairy cows. Given the difficulty in diagnosing C. burnetii infection at the farm level, an exhaustive series of tests in 43 pregnant animals that delivered at least one live calf were conducted, including blood serology and PCR of milk or colostrum, cotyledons (only at parturition), faeces, vaginal fluid against C. burnetii on gestation Day 171–177, at parturition and on Days 1–7, 8–14, 15–21, 22–28, 29–35 and 90–97 post-partum. During scheduled herd visits, ultrasonography (US) of the genital tract and examination of vaginal fluid were performed on Days 15–21 (V1), 22–28 (V2), 29–35 (V3) and 51–57 (V4) post-partum by the same veterinarian. Logistic regression analysis revealed that the likelihood of suffering endometritis (the presence of echogenic intrauterine fluid (IUF), cervical diameter of ≥4 cm or endometrial thickness ≥0.75 cm) was lower in C. burnetii-seropositive animals (OR = 0.10), compared with C. burnetii-seronegative animals. According to Kaplan–Meier survival analysis, C. burnetii-seronegative and non-shedding cows showed a delayed return to luteal activity and conception was delayed in non-shedding animals, compared with the remaining animals. Overall, the results of our study provide useful insight into the effects of C. burnetii infection on post-partum recovery and subsequent fertility. In particular, animals not infected with Coxiella seem to be susceptible to infection and not protected against the bacterium in dairy herds. The elevated costs of determining an infection at the farm level, make monitoring of cows virtually impossible from a clinical point of view.

The aim of this study was to evaluate the effect of body weight (BW) change during the early postpartum period and BW on day 50 post partum on the subsequent expression of estrus and fertility at first insemination before day 90 post partum. The data were derived from 1036 postpartum cows reaching 90 days in milk. The following data were recorded for each animal: lactation number, daily milk production at day 50 post partum, calving date, days in milk at estrus and at AI, insemination date, insemination number, inseminating bull, AI technician and body weight at calving, at day 50 post partum and at AI. Of the 1036 cows registered, 817 (78.9%) showed first estrus before day 90 post partum and were inseminated. Using logistic regression procedures and based on the odds ratio a one unit (kg) increase in the daily milk production at day 50 post partum and a one unit (kg) decrease in the BW between calving and day 50 post partum were related to a 1.03-fold increase and a 0.97-fold decrease in the estrus expression rate before day 90 post partum, respectively. Of the 817 cows inseminated before day 90 post partum, 437 became pregnant (53.5%). A one unit increase in the BW at day 50 post partum produced a 1.003-fold increase in the early fertility rate. Pregnancy rate before day 90 was also influenced by the season at calving, bull providing semen and technician. In conclusion, the register of BW during the postpartum period may be a useful tool for evaluating the nutritional status and its relationship with the subsequent reproductive efficiency in dairy cows.

This study sought to improve the reproductive performance of anoestrous high-producing dairy cows by including equine chorionic gonadotrophin (eCG) after progesterone-releasing intravaginal device (PRID) removal. In Experiment I, 806 cows at 51-57 days post-partum were randomly assigned to a PRID (treated with PRID), PRID-500 (treated with PRID plus 500 IU of eCG) or PRID-750 (treated with PRID plus 750 IU of eCG) group. In Experiment II, 422 cows showing a long anoestrus period (animals with no oestrus signs nor luteal tissue 35 days before treatment) were randomly assigned to the PRID, PRID-500 or PRID-750 groups. The dependent variables considered in binary logistic regression analyses for both experiments were the rates of oestrus, ovulation and conception after treatment, the cumulative conception rate on Day 120 post-partum and pregnancy loss. In Experiment I, interaction between treatment and season showed a significant effect on the oestrous response. Thus, during the warm season, PRID group cows were 8.9 times more likely to express oestrus than the remaining cows. Moreover, inseminated cows with two or more corpora lutea 8-14 days after treatment were more likely to become pregnant (by a factor of 2.4) than cows with a single corpus luteum. Finally, cows without luteal structures treated with PRID were 0.4 less likely to be pregnant on Day 120 post-partum, compared with the remaining cows. In Experiment II, cows in the PRID group treated during the warm or cool season were less likely to exhibit oestrus (by a factor of 0.06 or 0.2, respectively) or ovulate (by a factor of 0.004 or 0.14, respectively) than the remaining cows. In conclusion, in anoestrous cows in both experiments, the addition of eCG to the use of an intravaginal progesterone device to induce oestrus was beneficial. The recommended dose of eCG is 500 IU.

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, P0.001) 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.