Search

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.

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.

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.

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.

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.

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).

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

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 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.

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.

This study was designed to test the treatment with prostaglandin F2α (PGF2α) plus equine chorionic gonadotrophin (eCG) and GnRH 48 h later (PEG protocol) followed by fixed-time AI (FTAI) in dairy cows with silent ovulation (cows with a mature corpus luteum and no signs of estrus detected in the preceding 21 days). In Experiment I, ovulation following the PEG protocol monitored in 24 cows with silent ovulation was recorded in 83% of the cows 36 h after GnRH treatment. In Experiment II, control cows were artificially inseminated during spontaneous estrus (4615 AI), while cows in the PEG group (1266 AI) were subjected to FTAI 24 h after GnRH treatment. Binary logistic regression revealed a significant effect of the interactions of treatment by season, by parity or by repeat breeding syndrome (>3 AI) on the conception rate. The conception rate was negatively affected by the warm season and age in controls but not in treated cows, whereas repeat breeder cows in the control and PEG groups were less (by a factor of 0.7) or more (by a factor of 1.5) likely to become pregnant, respectively, than the remaining animals. Moreover, the likelihood of twin pregnancy was lower in multiparous PEG (by a factor of 0.4) cows than in the remaining cows. This protocol, besides overcoming the negative effects of heat stress and age on the conception rate, increased fertility in repeat breeder cows compared with spontaneous estrus. Moreover, this treatment regimen reduced the twin pregnancy rate in multiparous cows.

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.

This study was designed to determine the effects of twinning on the subsequent reproductive performance and productive lifespan of high milk-producing dairy cows. The data analyzed were taken from complete reproductive records for 4861 Holstein Friesian cows comprising a commercial herd, including data for 12 587 calving events from April 1, 2001 to March 31, 2012. The twinning rate was 5.6%, and 9.6% of the cows delivered twins at least once during their life. Conception rates before Days 90, 120, and 300 postpartum were 20.8%, 36.9%, and 74.5%, and 34.2%, 51.8%, and 85.0% for cows delivering twins and singletons, respectively. Cox regression analysis revealed that twinning reduced the chances of conception before Day 90 by a factor of 0.76. Also, the median calving to conception interval was significantly longer for cows calving twins (134 ± 4.5 days) than for cows delivering singletons (108 ± 0.8 days; P < 0.001). Moreover, in cows in which conception was successful, the risk of abortion was higher for cows that calved twins than for those calving singletons (13.7% vs. 10.3%, respectively; P = 0.01). Culling rates before Days 90, 120, and 300 postpartum were 15.6%, 16.1%, and 28.6%, and 7.6%, 8.7%, and 15.9% for cows calving twins and singletons, respectively. Cox regression analysis of the factors affecting the culling rate before Day 300 postpartum revealed a 1.41 times greater hazard ratio of culling for cows calving twins than cows calving singletons. Further, mean productive lifespan was almost 300 days shorter for primiparous twinners (N = 48, 602 ± 493 days) than for nontwinners (N = 2592; 899 ± 581 days; P < 0.01), and 200 days shorter for secundiparous twinners (N = 126, 914 ± 429 days) than for nontwinners undergoing at least two lactations (N = 1936, 1101 ± 522 days; P < 0.01). Kaplan-Meier survival curves for productive lifespan differed between primiparous twinners and non-twinners (P < 0.001), and between secundiparous twinners and nontwinners having at least two lactations (P = 0.017). Differences in culling patterns for twinners and nontwinners were not restricted to the subsequent lactation but continued as long as 800 days after first calving, strongly suggesting long-term negative effects of twinning. As an economic implication of our findings, we suggest that twin embryo reduction at the moment of pregnancy diagnosis could be a profitable strategy to cut twinning rates and abolish their detrimental effects on subsequent reproductive performance and productive lifespan.

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.

To avoid the problems associated with twinning in dairy cattle, one of the embryos may be eliminated. This study compares the effect on pregnancy maintenance of two embryo reduction techniques, manual rupture (MR) and transvaginal ultrasound-guided aspiration (TUGA) of allanto-amniotic fluid, in Holstein-Friesian cows with multiple pregnancies. In the first experiment, 61 lactating cows bearing unilateral twins (n=27), bilateral twins (n=30) or triplets/quadruplets (n=4) were subjected to MR (n=45) or TUGA using a 17-G neddle (n=16) on day 28-34 of gestation. In 21 and 10 cows undergoing MR and TUGA embryo reduction, respectively, pregnancy loss occurred before day 90 (46.7 vs. 62.5%, P= 0.28). Through binary logistic regression, the type of pregnancy was identified as the only variable significantly affecting pregnancy maintenance (P=0.03). Based on the odds ratio, the risk of pregnancy loss was 4.1 times higher for unilateral twins than for bilateral twins (70.4 vs. 36.7%, respectively, P=0.01). No effect was detected on pregnancy maintenance of the technique used (P=0.17) or of the interaction technique by type of pregnancy (P=0.22). In the second experiment, a 22-G needle was used to perform TUGA on 22 lactating cows. The pregnancy loss rates were 44.4% (4/9), 18.2% (2/11) and 50% (1/2) for cows bearing unilateral twins, bilateral twins and triplets, respectively. The total pregnancy loss rate following TUGA using the 22-G needle tended to be lower than that using the 17-G needle (31.8 vs. 62.5%; P=0.06). Our results suggest that TUGA using a 22-G needle could be the method of choice to perform embryo reduction in cows carrying multiple pregnancies.

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.

This study was designed to test the effects of progesterone or GnRH treatment on day 5 post-AI on fertility and luteal function in dairy cows and heifers. Five days after AI, 32 animals were randomly assigned to a control, intravaginal progesterone for 14 days progesterone releasing intravaginal device (PRID) or GnRH treatment group. On days 5, 7, 12, 14, 17 and 19 post-AI, each animal underwent colour Doppler ultrasonography of the corpus luteum and blood samples were collected for cell counts and plasma progesterone determination. Through general linear model repeated measures analysis of variance, significant effects were observed of treatment, parity, inseminating bull, reduced vascularization of the CL and pregnancy on plasma progesterone concentrations, whereas mean plasma progesterone and time luteal phase day, and treatment and plasma progesterone concentration on day 5 post-AI were found to, respectively, affect neutrophil and lymphocyte counts throughout the luteal phase. Moreover, two binary logistic regression analyses were performed. Based on the odds ratio, the likelihood of pregnancy by days 26-32 post-AI was 23.4 times higher in animals with high mean progesterone levels throughout the study period, compared with animals with low mean progesterone. The likelihood of reduced CL vascularization was 14 times higher in animals treated with PRID, compared with control and GnRH-treated animals. In conclusion, our results indicate that treatment on day 5 post-AI with PRID reduced subsequent CL vascularization, whereas GnRH treatment increased plasma progesterone concentrations on day 12 post-AI, although an effect was identified of the inseminating bull on plasma progesterone levels. Pregnant animals showed higher mean plasma progesterone concentrations than in nonpregnant ones and heifers higher than in lactating cows, whereas blood cell counts differed depending on the treatment and on the mean plasma progesterone concentration on day 5 post-AI.

Contents The aims of this study were to analyse, in high-producing dairy cows, plasma Coxiella burnetii antibody titres and seroconversion throughout gestation, along with possible factors affecting such titres. The study was performed on 65 lactating pregnant non-aborting animals in a commercial Holstein-Friesian dairy herd in northeastern Spain. Blood samples for antibody determinations were collected on days 40, 90, 120, 150, 180 and 210 of gestation. By General Linear Model (GLM) repeated measures analysis of variance, the effects of milk production and reproductive variables as well as Neospora caninum-seropositivity on C. burnetii antibody levels for all animals and for seropositive animals were established. Significant effects were observed of day of gestation, parity and N. caninum-seropositivity (between subject effects) on the C. burnetii antibody levels recorded for the whole population of animals throughout the gestation period. C. burnetii antibody levels were higher in primiparous than in multiparous cows, with titres in primiparous cows diminishing during the post-partum period. In seropositive cows, significant effects were observed of milk production and inseminating bull on gestational C. burnetii antibody levels. When the data were subjected to binary logistic regression considering C. burnetii-seropositivity as the dependent variable, the resultant odds ratios indicated that the likelihood of C. burnetii-seropositivity was lower in N. caninum-seropositive animals (OR 0.12) compared to N. caninum-seronegative animals, and in multiparous cows (OR 0.12) compared to primiparous cows. In conclusion, Coxiella-infected dams remained seropositive during the whole gestation period, though primiparous cows showed a drop in antibody titres post-partum. No seronegative cow suffered seroconversion. Presence of both, N. caninum and C. burnetii antibodies in the same animal, was associated with a decrease in antibody titres against C. burnetii, perhaps indicating some cross-protection in animals infected by both pathogens.

This report reviews the clinical applications of routine ultrasound in gestation control programmes for dairy herds. Emphasis is placed on identifying risks of late embryo/early foetal loss of non-infectious cause by monitoring ovarian structures (as opposed to uterine contents), indicators of pregnancy loss and twin pregnancies. Suggestions are provided to appropriately manage a cow at risk of pregnancy loss during the late embryonic/early foetal period. This review identifies gaps in our current knowledge of the aetiology of pregnancy loss and spontaneous embryo/corpus luteum reduction. Two areas of the implantation-placentation window of gestation in particular need urgent attention: the roles in pregnancy loss of immune-endocrine interactions and the physiological status of the embryo/corpus luteum. The latter issue and consecutively endocrine interactions could be successfully addressed by incorporating Doppler ultrasonography into routine ultrasound procedures.

Human chorionic gonadotropin (hCG) has a potent luteinizing hormone (LH)-like effect in cattle that extends the life span of the corpus luteum (CL) and increases progesterone synthesis, induces ovulation throughout the estrous cycle, promotes the formation of accessory corpora lutea when applied in the early luteal phase, and modifies follicular wave dynamics increasing the frequency of three-wave dominant follicular cycles. As hCG acts on ovarian cells independently of the pituitary gland and its effect is longer lasting than that produced by endogenous LH release, use of hCG rather than gonadotropin-releasing hormone (GnRH) could be targeted at populations of subfertile cows. This review describes the clinical use of hCG to improve the reproductive performance of dairy cows. In addition, we describe recent developments in the therapeutic use of hCG and studies addressing the benefits of including hCG in estrus and ovulation synchronization protocols. Our review ends with a critical discussion of how earlier findings related to ovarian responses to hCG treatment can be interpreted in the light of recent advances in the clinical applications of hCG.

Spontaneous reduction of advanced twin embryos has been described in high-producing, Holstein-Fresian (Bos taurus) dairy herds. The first objective of the current study was to determine whether management and cow factors could have an effect on such a reduction in twin pregnancies during the early fetal period. Because loss of a corpus luteum was noted in cows suffering twin reduction, we expanded our study to include multiple-ovulating cows carrying singletons. Pregnancy was diagnosed and confirmed from Days 28 to 34 and 56 to 62 postinsemination. Sixty-nine (23.5%) of 293 pregnant cows with two corpora lutea carrying singletons and 132 (28.4%) of 464 twin pregnancies recorded on first pregnancy diagnosis subsequently lost one of the corpora lutea or one of the embryos, respectively. Thirty-four (25.8%) of the 132 twin pregnancies suffering embryo reduction lost one corpus luteum along with the embryo. Corpus luteum reduction always occurred in the ovary ipsilateral to the gravid horn suffering embryo reduction. Binary logistic regressions were performed considering corpus luteum and embryo reduction as dependent variables in single and twin pregnancies, respectively, and several management- and cow-related factors as independent variables. In cows carrying singletons, the risk of corpus luteum reduction was 14.3 (1/0.07) times lower for a given herd, whereas the interaction season by laterality significantly affected corpus luteum reduction such that in cows with two corpora lutea ipsilateral to the horn of pregnancy, the risk of reduction decreased during the winter period. In cows carrying twins, ipsilateral twin pregnancies were 3.45 (1/0.29) times more likely to undergo the loss of one embryo than bilateral twin pregnancies. As an overall conclusion, both corpora lutea and embryos were vulnerable to the effects of stress factors during the early fetal period in cows maintaining their pregnancies. A strong unilateral relationship between the corpus luteum and the conceptus was also observed.

Contents The problem of pregnancy loss during the late embryonic/early foetal period due to non-infectious causes in dairy cattle is on the rise, especially in high milk producing cows. This review discusses recent therapeutic approaches designed to reduce the incidence of early foetal loss. These strategies are based on the fact that low plasma concentrations of progesterone are commonly related to foetal loss. Progesterone supplementation during the late embryonic/early foetal period has proven useful in curtailing losses, mainly in single pregnancies, whereas a more positive effect of treatment with GnRH than progesterone has been found in twin pregnancies. Therapeutics is not necessary in cows with an additional corpus luteum. In practical terms, suggestions include recording the number of embryos and corpora lutea on first pregnancy diagnosis and checking for normal pregnancy progression on day 60 post-insemination.

Contents The aim of this study was to investigate the effect of applying a progesterone-based oestrous synchronization protocol at 51–57 days postpartum in high-producing dairy cows. The data analysed were derived from 1345 lactating cows. Cows between 51 and 57 days postpartum were assigned to the groups: control, PRID (receiving a progesterone-releasing intravaginal device for 9 days, and prostaglandin F2α 24 h before PRID removal) or GnRH–PRID (the same as the PRID group plus GnRH at PRID insertion). Oestrus was detected by using pedometers and confirmed by examination of the genital tract at AI. Oestrous and conception rates before days 71–77 postpartum, pregnancy loss in early pregnant cows or the cumulative conception rate registered on day 120 postpartum were considered as the dependent variables in four consecutive logistic regression analyses. Based on the odds ratios, the oestrous rate increased by a factor of 1.73 in cows showing oestrus before treatment for each unit increase in the number of previous oestruses; decreased by a factor of 0.44 in the control group with respect to the treatment groups; and by a factor of 0.61 in cows without luteal structures at treatment with respect to cows with corpora lutea. The conception rates of cows inseminated before days 71–77 postpartum remained similar across the groups, whereas the likelihood of pregnancy loss for cows becoming pregnant during this period was 0.11 times lower in the PRID group than in the control. Based on the odds ratio, the likelihood of a higher cumulative conception rate on day 120 postpartum: increased in cows showing oestrus before treatment by a factor of 1.41 for each unit increase in the number of previous oestruses, was reduced 0.56-fold in control cows compared with treated cows, and was also reduced by a factor of 0.98 for each kilogram of milk production increase recorded at treatment. In conclusion, although oestrous synchronization programmes performed in this study did not improve fertility, cows treated with progesterone could be inseminated earlier than untreated cows, such that the treatments increased the cumulative pregnancy rates determined on day 120 postpartum. In addition, fewer pregnancy losses were observed in early pregnant cows in the PRID group than the GnRH–PRID group.

This study was designed to assess the reproductive performance of Neospora caninum-infected dairy cows (Bos taurus) after abortion. The population examined was composed of 92 aborting cows subsequently subjected to at least one artificial insemination (AI) attempt. Of these animals, 68 were N. caninum seropositive and 24 were seronegative. Only animals producing at least 25 kg milk at the estrus after abortion were inseminated. Normal uterine involution was verified at first AI after abortion in cows showing estrous signs within 30 d of abortion. Of the 92 aborting cows, 73 (79.3%) became pregnant after abortion: 57 (83.8%) and 16 (66.7%) of the N. caninum seropositive and seronegative cows, respectively. The factors days of gestation and lactation number at abortion failed to differ between the two Neospora serostatus groups, based on Student's t-test or Kruskal-Wallis tests, respectively. The interval from abortion to first AI, the number of AIs necessary for an animal to become pregnant, and the interval from abortion to next pregnancy in cows becoming pregnant were lower for the N. caninum-seropositive than for the N. caninum-seronegative cows (P

This study was designed to establish the factors, if any, which could affect plasma pregnancy-associated glycoprotein-1 (PAG-1) expression in a study population of 87 pregnant, high-producing dairy cows. The factors examined were: semen providing breed (Holstein-Friesian vs Limousin), outcome of gestation (male vs female newborn, and singleton vs twin pregnancies), lactation number, milk production at pregnancy diagnosis, plasma progesterone concentration, season of gestation (warm period, March-November vs cool period, December-February), and day of gestation (40, 90, 120, 150, 180 and 210). Pregnancy was diagnosed by transrectal ultrasound on day 40 post-insemination and by palpation per rectum on days 90, 120, 150, 180 and 210. Blood samples were collected from each animal immediately before each pregnancy diagnosis. The relative contributions of the different factors on PAG-1 concentrations were evaluated by GLM repeated measures analysis of variance. No significant effects of the herd, foetal sex, milk production, lactation number and plasma progesterone concentrations were observed. In contrast, twin pregnancy, the use of Limousin semen and conception during the cool period were correlated with significantly increased plasma PAG-1 concentrations throughout gestation. Our data indicate that both cow well-being during early placental development, determined in our conditions by reduced heat stress when conception occurred in the cool season, and crossbreed pregnancies lead to improved PAG-1 production throughout the gestation period.

This study analyses anomalous cases of gestation ending in pregnancy loss during the early foetal period and their effect on progesterone and plasma pregnancy-associated glycoprotein-1 (PAG-1) concentrations. Data derived from a large-scale ultrasound pregnancy diagnosis programme in high producing dairy cows. Over a 3-year period (2004-2007), a very low incidence (0.5%: 15 of 3094) of anomalous pregnancies was recorded. The results revealed that the following anomalies were detected on days 35-41 of gestation in cows carrying singletons with one single corpus luteum: embryo death in eight cows (0.3%); and embryo in the uterine horn contralateral to the corpus luteum in seven cows (0.2%). All these animals suffered pregnancy loss during the early foetal period. In cows carrying dead embryos, no signs of conceptus degeneration were observed on pregnancy diagnosis. Amnion size (approximately 25 mm diameter) and uterine horn fluid contents were estimated to be similar to those of the normal pregnant cows in this period. In the contralateral gestations, live embryos were observed in all ultrasound checks before pregnancy loss. Uterine fluid contents increased in the two cows in which gestation continued for more than a week. In the cases of embryo death but not in those of contralateral gestation, a drop in PAG-1 levels was noted prior to pregnancy loss. Two cows carrying dead embryos increased with time allantoic fluid contents. The PAG-1 values increased with time in one cow bearing a dead embryo (from 2.31 to 6.79 ng/ml) and in two of the contralateral gestations (from 1.66 to 2.33 ng/ml and from 0.39 to 6.79 ng/ml, respectively). Results of this study indicate that the foetal membranes continue to undergo some activity following embryo death, and that contralateral pregnancy may determine failure of the gestation process.

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

The aim of the present study was to investigate possible relationships between plasma concentrations of prolactin and the following factors throughout gestation in lactating dairy cows: photoperiod, season, milk production, Neospora caninum -seropositivity, twin pregnancy, and plasma concentrations of pregnancy associated glycoproteins-1 (PAG-1), progesterone and cortisol. Pregnancy was diagnosed by trans-rectal ultrasonography on Day 40 post-insemination and by palpation per rectum on Days 90, 120, 150, 180 and 210 or until abortion in aborting cows. Blood samples were collected from each animal immediately before each pregnancy diagnosis. The study population was comprised of 73 non-aborting (54 Neospora- seropositive cows) and 20 aborting cows (all Neospora- seropositive) cows. In non-aborting cows, GLM repeated measures of analysis of variance revealed that lactation number and days in milk had no effect on plasma prolactin concentrations throughout gestation, whereas high plasma prolactin concentrations were significantly associated with high plasma levels of cortisol and PAG-1, with Neospora -seropositivity, positive photoperiod (increasing day length), twin pregnancy, and with low plasma progesterone concentrations. An interaction among plasma prolactin, plasma cortisol and milk production was observed in that plasma prolactin concentrations differed significantly throughout gestation and were highest in high-producing cows with high cortisol levels. In Neospora -seropositive non-aborting versus aborting cows, mean prolactin concentrations failed to differ on Days 40, 90, 120, 150 and 180 of pregnancy, whereas prolactin values were significantly lower ( P =0.03) in aborting animals on Day 210 of pregnancy. Our results indicate that a positive photoperiod and Neospora -infection lead to increased plasma prolactin concentrations throughout gestation. Reduced prolactin concentrations detected in Neospora -seropositive aborting cows compared to non-aborting animals suggests a protective effect of prolactin in N. caninum infection.

In the dairy industry worldwide, reproductive disorders are a major cause of economic losses and a challenge to scientists and technicians. In recent decades, declining fertility and increasing milk production have been widely reported in dairy cattle. In this article, the relationships between milk production, ovarian disorders and fertility in high-producing dairy herds are briefly described. We carried out a retrospective study of 23 204 lactations included in a reproductive control programme in north-eastern Spain, a geographical area experiencing both warm and cool conditions. The data were collected between 1991 and 2007 and refer to cows first inseminated or examined 45-80 days postpartum in five well-managed, commercial, Holstein-Friesian high-producing dairy herds. Ovarian disorders were classified as ovarian inactivity or hypofunction, cystic ovarian disease, sub-oestrus or silent ovulation and sub-luteal function. Ovarian hypofunction and milk production increased throughout the study period and there was a decrease in the pregnancy rate to first artificial insemination (AI). Cows suffering ovarian hypofunction were efficiently treated using combined progestagen-prostaglandin treatments. The incidence of ovarian cysts showed little variation with time. Treatment of this syndrome may include different GnRH-based treatments or manual rupture. During the last 5 years, sub-oestrus was the predominant dysfunction (42.1%) compared with the cystic (6.3%) and ovarian hypofunction (12%) forms. Response of sub-oestrous cows to treatment with luteolitic agents was usually higher than 60%. Ovarian function and fertility were dramatically impaired during the warm period. However, during the later years of the study, the inclusion of fans and water sprinklers for the warm season appeared to overcome the seasonal effect on fertility.

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