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Simple Summary: Heat stress is a major problem for the health and productivity of livestock, including cattle. Since the 1950s, researchers have studied how high environmental temperature affects cattle fertility through both laboratory and field studies. When a cow's rectal or vaginal temperature goes above 39.1 °C, it is considered heat-stressed, which can harm its ability to reproduce. However, there is not enough information about temperatures in the uterus, oviducts, and ovarian follicles, especially in lactating dairy cows, to ensure current laboratory models are accurate. These data are crucial for understanding how heat stress impacts oocyte (egg) development and early embryo survival in cattle. Additionally, it is important to improve live animal stress models to correctly identify when cows are truly experiencing heat stress, shown by signs like increased panting, body temperature, and heart rate, rather than just assuming they are stressed based on exposure to a high temperature–humidity index. Improving these models will make them more reliable and help identify cattle that can better tolerate heat. Rectal and vaginal temperatures are utilised in both in vivo and in vitro models to study the effects of heat stress on oocyte competence and embryo viability in cattle. However, uterine temperature increases by only 0.5 °C in heat-stressed cows, significantly lower than simulated increases in in vitro models. Temperature variations within oviducts and ovarian follicles during heat stress are poorly understood or unavailable, and evidence is lacking that oocytes and pre-implantation embryos experience mild (40 °C) or severe (41 °C) heat stress inside the ovarian follicle and the oviduct and uterus, respectively. Gathering detailed temperature data from the reproductive tract and follicles is crucial to accurately assess oocyte competence and embryo viability under realistic heat stress conditions. Potential harm from heat stress on oocytes and embryos may result from reduced nutrient availability (e.g., diminished blood flow to the reproductive tract) or other unidentified mechanisms affecting tissue function rather than direct thermal effects. Refining in vivo stress models in cattle is essential to accurately identify animals truly experiencing heat stress, rather than assuming heat stress exposure as done in most studies. This will improve model reliability and aid in the selection of heat-tolerant animals. [ABSTRACT FROM AUTHOR]

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

Simple Summary: The impact of heat stress on reproduction is complex and multifactorial and is compounded by growing challenges due to climate change. Both animal welfare and fertility are vulnerable parameters easily affected by heat stress. Heat stress leads to a marked decrease in the developmental competence of oocytes and the fertilizing capacity of spermatozoa, leading to a declining reproduction rate and losses for the cattle industry. There is an urgent need to find viable methods of alleviating the effects of global warming on animal welfare and fertility and, accordingly, guard the reproductivity, productivity, and profitability of cattle farms. In recent decades, the adverse effects of global warming on all living beings have been unanimously recognized across the world. A high environmental temperature that increases the respiration and rectal temperature of cattle is called heat stress (HS), and it can affect both male and female reproductive functions. For successful reproduction and fertilization, mature and healthy oocytes are crucial; however, HS reduces the developmental competence of oocytes, which compromises reproduction. HS disturbs the hormonal balance that plays a crucial role in successful reproduction, particularly in reducing the luteinizing hormone and progesterone levels, which leads to severe problems such as poor follicle development with a poor-quality oocyte and problems related to maturity, silent estrus, abnormal or weak embryo development, and pregnancy loss, resulting in a declining reproduction rate and losses for the cattle industry. Lactating cattle are particularly susceptible to HS and, hence, their reproduction rate is substantially reduced. Additionally, bulls are also affected by HS; during summer, semen quality and sperm motility decline, leading to compromised reproduction. In summer, the conception rate is reduced by 20–30% worldwide. Although various techniques, such as the provision of water sprinklers, shade, and air conditioning, are used during summer, these methods are insufficient to recover the normal reproduction rate and, therefore, special attention is needed to improve reproductive efficiency and minimize the detrimental effect of HS on cattle during summer. The application of advanced reproductive technologies such as the production of embryos in vitro, cryopreservation during the hot season, embryo transfer, and timed artificial insemination may minimize the detrimental effects of HS on livestock reproduction and recover the losses in the cattle industry. [ABSTRACT FROM AUTHOR]

Contents: 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. [ABSTRACT FROM AUTHOR]

The negative effects of heat stress on dairy cattle's fertility have been extensively studied, but the relevant knowledge for beef cattle is rather limited. The aims of this study were to investigate the effects of HS during in vitro maturation on the developmental potential of oocytes derived from Limousine and Holstein cows and to estimate the effect of the differential gene expression of important genes in oocytes, cumulus cells and blastocysts in the growth competence between the breeds. In seven replicates, cumulus oocyte complexes from Holstein and Limousine cows were matured for 24 hr at 39°C (controls C; Hol_39, Lim_39) or at 41°C from hour 2 to hour 8 of IVM (treated T; Hol_41, Lim_41), fertilized, and presumptive zygotes were cultured for 9 days at 39°C. Cleavage and embryo formation rates were evaluated 48 hr post‐insemination and on days 7, 8 and 9, respectively. From all groups, subsets of cumulus cells, oocytes and blastocysts were analysed for the relative expression of genes related to metabolism, stress, apoptosis and placentation. No difference was detected in cleavage rate or in blastocyst formation rate among the control groups. In both breeds, heat stress reduced blastocyst yield, but at all days the suppression was higher in Limousines. In Holsteins, altered gene expression was detected in cumulus cells (G6PD, GLUT1) and blastocysts (PLAC8), while in Limousines, differences were found in oocytes (G6PD, HSP90AA1), in cumulus cells (CPT1B, HSP90AA1, SOD2) and blastocysts (DNMT, HSP90AA1, SOD2). It appears that Holstein COCs are more tolerant than Limousine COCs, possibly due to compulsory, production driven selection. [ABSTRACT FROM AUTHOR]

Summary: To investigate the effects of physiologically relevant heat shock during oocyte maturation, buffalo cumulus–oocyte complexes (COCs) were cultured at 38.5°C (control) or were exposed to 39.5°C (T1) or 40.5°C (T2) for the first 6 h of in vitro maturation (IVM), followed by 38.5°C through the next 18 h/IVM and early embryonic development up to the blastocyst stage. Gene expression analysis was performed on selected target genes (HSF-1, HSF-2, HSP-70, HSP-90, BAX, p53, SOD1, COX1, MAPK14) in denuded oocytes and their isolated cumulus cells resulting from control COCs as well as from COCs exposed to a temperature of 39.5°C (T1). The results indicated that heat shock significantly (P < 0.01) decreased the maturation rate in T1 and T2 cells compared with the control. After in vitro fertilization (IVF), cleavage rate was lower (P < 0.01) for oocytes exposed to heat stress, and the percentage of oocytes arrested at the 2- or 4-cell stage was higher (P < 0.01) than that of the control. The percentage of oocytes that developed to the 8-cell, 16-cell or blastocyst stage was lower (P < 0.01) in both T1 and T2 groups compared with the control group. mRNA expression levels for the studied genes were decreased (P < 0.05) in treated oocytes (T1) except for HSP-90 and HSF-1, which were increased. In cumulus cells isolated from COCs (T1), the expression for the target genes was upregulated except for BAX, which was downregulated. The results of this study demonstrated that exposure of buffalo oocytes to elevated temperatures for 6 h severely compromised their developmental competence and gene expression. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Contents 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. [ABSTRACT FROM AUTHOR]

Heat stress (HS) is especially harmful for bovine ovarian follicle development and oocyte competence. Furthermore, HS causes premature aging in oocytes due to high levels of reactive oxygen species (ROS), involved in the harmful effects over the oocyte maturation and the steroidogenic activity of follicular cells. In this study, the presumptive protective effects of antioxidant agents on heat-stressed oocytes were evaluated. Heifer oocytes were matured for 22 h under control (38°C) and HS conditions (41.5°C at 18–21 h of maturation). For each oocyte, nuclear stage and cortical granule (CG) distribution were evaluated. Steroidogenic activity of cumulus cells was also recorded. The antioxidant agents used in the study were: retinol (1.43 μg/ml), retinyl (0.28 μg/ml) and oleic acid (0.05 mg/ml). Based on a chi-squared test (P < 0.05), HS affected negatively the metaphase II (MII) progression and produced a premature CG exocytosis. Retinol improved the oocyte MII progression. However, retinyl and oleic acid, at the concentrations used in this study, could not counteract adverse effects of HS. A decrease in progesterone and increase in estradiol availability were observed when retinyl and oleic acid were supplemented to the maturation medium, respectively. In conclusion, retinol proved to be valuable in heat-stressed oocytes protecting nuclear maturation. [ABSTRACT FROM PUBLISHER]

Simple Summary: The Holstein cows are among the most thermosensitive farm animals. In this breed, during the heat stress periods, fertility is seriously compromised due to induced alterations of the endocrine status, reduced fertilizing capacity of the oocyte and increased embryo deaths. To combat the deleterious effects of stress, cells synthesize a series of specific molecules that are mainly involved in cellular protection against the heat insult, called heat shock proteins (HSPs). Here, we examined the effects of supplementing HSP70 in in vitro matured bovine oocytes under thermoneutral or heat stress conditions, and we assessed its efficacy on in vitro embryo yield and quality; the latter was determined on the basis of the expression of various genes related to important cellular functions. It was manifested that HSP70 addition into the in vitro maturation medium restores the developmental competence of heat stressed oocytes and improves the quality of the in vitro produced embryos. Heat shock protein 70 (HSP70) is a chaperon that stabilizes unfolded or partially folded proteins, preventing inappropriate inter- and intramolecular interactions. Here, we examined the developmental competence of in vitro matured oocytes exposed to heat stress with or without HSP70. Bovine oocytes were matured for 24 h at 39 °C without (group C39) or with HSP70 (group H39) and at 41 °C for the first 6 h, followed by 16 h at 39 °C with (group H41) or without HSP70 (group C41). After insemination, zygotes were cultured for 9 days at 39 °C. Cleavage and embryo yield were assessed 48 h post insemination and on days 7, 8, 9, respectively. Gene expression was assessed by RT-PCR in oocytes, cumulus cells and blastocysts. In C41, blastocysts formation rate was lower than in C39 and on day 9 it was lower than in H41. In oocytes, HSP70 enhanced the expression of three HSP genes regardless of incubation temperature. HSP70 at 39 °C led to tight coordination of gene expression in oocytes and blastocysts, but not in cumulus cells. Our results imply that HSP70, by preventing apoptosis, supporting signal transduction, and increasing antioxidant protection of the embryo, protects heat stressed maturing bovine oocyte and restores its developmental competence. [ABSTRACT FROM AUTHOR]

Simple Summary: In dairy herds, twin pregnancies considerably compromise cow welfare, reducing their lifespan and leading to the increased use of antibiotics postpartum. In cows with follicles of pre-ovulatory size at insemination, twins can be avoided by puncturing and draining one follicle. In this report, we propose the use of a recently described manual method of follicular drainage without the need for ultrasound guidance and its association with treatment of the animal with human chorionic gonadotrophin (hCG), thus also increasing fertility compared to the use of Gonadotrophin releasing hormone (GnRH). The manual follicular drainage technique is simple, takes less than 1 min, and can be performed by a clinical veterinarian. In this study, even during heat stress periods follicle drainage plus hCG treatment gave rise to nearly 50% of pregnancies relative to conception rates, compared to 29% observed in non-drained, non-treated control animals or 20% in animals drained and treated with GnRH. This approach is recommended for weekly reproductive program visits. Twin pregnancies are undesirable in dairy cattle as they dramatically compromise cow lifespan and, consequently, herd economy. Clinical problems in cows arise from the time of pregnancy diagnosis to pregnancy loss, abortion, or parturition. The drainage of co-dominant follicles in cows with two or more follicles at insemination prevents twin pregnancy. The objective of this study was to compare the effectiveness of draining the smaller of two follicles through a simplified maneuver not requiring ultrasonography in cows in their third or more lactation, and then inducing ovulation immediately before artificial insemination (AI) with GnRH or human chorionic gonadotrophin (hCG). Animals were monitored by ultrasound at AI and randomly assigned to the groups: follicular drainage and treatment with GnRH (Deph; n = 60), follicular drainage and treatment with hCG (hCG; n = 60), and non-drainage (ND; n = 60) as control cows. On the basis of odds ratios, cows treated with hCG were 2.1 times more likely to become pregnant than control animals. Our results reveal the efficacy of hCG treatment at AI in cows with two follicles of pre-ovulatory size subjected to a simple follicular drainage procedure. [ABSTRACT FROM AUTHOR]

Simple Summary: Cows have two ovaries, two uterine horns, two oviducts, and a uterine body. Although they are commonly monovular, they can sustain twin or other multiple pregnancies to term. In cows with a single pregnancy, the embryo implants in the uterine horn that is on the side of ovulation where a corpus luteum develops. Multiple pregnancies are classified into bilateral (one or more embryos in each uterine horn) and unilateral (all embryos in the same uterine horn, right or left) and, in both cases, embryos occur on the side of their corresponding corpus luteum or corpora lutea. Multiple pregnancies are undesirable in dairy herds as they compromise the reproductive performance and productive lifespan of cows. The present study sought to: (a) gain information on the incidence of twin pregnancies, (b) assess and expand information on the relative incidence of bilateral twins compared to unilateral twins, (c) confirm corresponding laterality between embryos and corpora lutea, (d) tabulate the frequency of triplets or quadruplets, and (e) evaluate possible effects of environmental heat stress conditions on the incidence of multiple pregnancies. The data for this study were derived from the ultrasonographic examination of 1130 cows carrying twins, triplets, or quadruplets, and 3160 cows carrying singletons. 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 < 0.0001) in the conception rate (28–34 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 < 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 < 0.0001). Thus, a warm climate is the main factor compromising the fate of multiple pregnancies. Some clinical suggestions are provided. [ABSTRACT FROM AUTHOR]