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Widespread genotyping has enabled the identification of putative recessive mutations that affect fertility through early embryonic fetal loss, or that compromise neonate or calf viability. The use of artificial insemination in the global dairy population can rapidly spread these harmful mutations, and testing for multiple mutations can become relatively expensive if not all tests are available on the same SNP panel. However, it is possible to provide heifer and cow predicted carrier status to farmers at no additional cost if the animals are genotyped with a standard SNP panel. Additionally, for defects where the causal mutation is unknown but a haplotype of markers has been associated with the defect, the carrier status can be predicted based on that haplotype. The aims of this study were 3-fold: (1) to determine the accuracy of imputation of putative causal mutations for recessive deleterious conditions in Australian dairy cattle, (2) to impute carrier status for known recessive deleterious conditions in all genotyped Australian Holstein, Jersey, and Red breed cows, and (3) to determine the changes in carrier frequencies across time for these recessive deleterious mutations. We used the F1 statistic, combining precision and recall, to assess the accuracy of carrier status prediction. We showed that known deleterious mutations can be accurately imputed in Australian Holstein and Jersey cattle that are not directly genotyped for the causal mutation, with F1 ranging between 0.88 and 0.99. For recessive deleterious conditions not included on the standard Australian SNP panel, carrier status could be predicted using a marker haplotype, with F1 ranging from 0.91 to 0.92. Most putative causals and haplotypes were either stable with a low carrier percentage or had a declining carrier percentage. However, several recessive mutations showed a relatively high or increasing percentage, highlighting the importance of detecting carriers to reduce the number of at-risk matings. Furthermore, the high carrier percentage of the recently identified bovine lymphocyte intestinal retention defect mutation emphasizes the importance of detection of novel mutations., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Heifer conception rate to the first service (HCR1) is defined as the number of heifers that become pregnant to the first breeding service compared to the heifers bred. This study aimed to identify loci associated and gene sets enriched for HCR1 for heifers that were bred by artificial insemination (AI, n = 2829) or were embryo transfer (ET, n = 2086) recipients, by completing a genome-wide association analysis and gene set enrichment analysis using SNP data (GSEA-SNP). Three unique loci, containing four positional candidate genes, were associated ( p < 1 × 10 -5 ) with HCR1 for ET recipients, while the GSEA-SNP identified four gene sets (NES ≥ 3) and sixty-two leading edge genes (LEGs) enriched for HCR1. While no loci were associated with HCR1 bred by AI, one gene set and twelve LEGs were enriched (NES ≥ 3) for HCR1 with the GSEA-SNP. This included one gene ( PKD2 ) shared between HCR1 AI and ET services. Identifying loci associated or enriched for HCR1 provides an opportunity to use them as genomic selection tools to facilitate the selection of cattle with higher reproductive efficiency, and to better understand embryonic loss.

A greater understanding of factors influencing fertility is essential to improve pregnancy rates and reduce the occurrence of embryonic mortality in beef herds. The objective of the current study was to evaluate retrospective data of pregnancy per artificial insemination (P/AI) and pregnancy loss in Nelore females subjected to timed-AI (TAI) in Brazil. Data from 40,104 TAI collected from six breeding seasons (2016-2022) were analyzed, and the effects of animal category (e.g., classification based on age and parity), farm, month of parturition, sire, sire breed (Nelore vs Angus), estrus expression at TAI, animal temperament, and body condition scores (BCS) were evaluated. P/AI and pregnancy loss were affected (P < 0.001) by animal category. There was also an effect of farm (P = 0.0013) on P/AI and pregnancy loss (P = 0.001), as P/AI ranged from 49.28% and 55.58% and pregnancy loss from 3.37% to 6.89% across the herds evaluated. Month of parturition also affected (P < 0.001) P/AI and was higher for cows that became pregnant at the beginning of the previous breeding season. Calmer animals, presenting lower velocity scores while exiting the chute following TAI, achieved higher P/AI (P < 0.001). Lower BCS at TAI was associated (P < 0.001) with increased pregnancy loss, and BCS gain following AI was associated (P < 0.001) with reduced rates of embryonic mortality. There was a major effect (P < 0.001) of sire on P/AI and pregnancy loss, as P/AI ranged from 11% to 79%, and embryonic mortality from 0% to 40% for the bulls used in the study, highlighting the importance of the sire fertility on overall pregnancy success. Results from the current study reinforce the idea that animal age and parity at the beginning of the breeding season, BCS at the onset of estrous synchronization, BCS gain following AI, estrus expression at TAI, sire, and month of parturition are important factors influencing P/AI and rates of embryonic mortality in beef herds., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

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

A 60% pregnancy success for inseminations is targeted to optimize production efficiency for dairy cows within a seasonal, pasture-grazed system. Routine measures of pregnancy success are widely available but are limited, in practice, to a gestation stage beyond the first 28 d. Although some historical data exist on embryonic mortality before this stage, productivity of dairy systems and genetics of the cows have advanced significantly in recent decades. Accordingly, the aim was to construct an updated estimate of pregnancy success at key developmental stages during the first 70 d after insemination. Blood samples were collected for progesterone concentrations on d 0 and 7. A temporal series of 4 groups spanning fertilization through d 70 were conducted on 4 seasonal, pasture-grazed dairy farms (n = 1,467 cows) during the first 21 d of the seasonal breeding period. Morphological examination was undertaken on embryos collected on d 7 (group E7) and 15 (group E15), and pregnancy was diagnosed via ultrasonography on approximately d 28 and 35 (group E35) as well as d 70 (group E70). Fertilization, embryo, and fetal evaluation for viability established a pregnancy success pattern. Additionally, cow and on-farm risk factor variables associated with pregnancy success were evaluated. We estimated pregnancy success rates of 70.9%, 59.1%, 63.8%, 62.3%, and 56.7% at d 7, 15, 28, 35, and 70, respectively. Fertilization failure (15.8%) and embryonic arrest before the morula stage (10.3%) were the major developmental events contributing to first-week pregnancy failures. Embryo elongation failure of 7% contributed to pregnancy failure during the second week. The risk factors for pregnancy success that were related to the cows included interval between calving and insemination, and d-7 plasma progesterone concentrations, whereas insemination sire was associated with pregnancy outcome. Most pregnancy failure occurs during the first week among seasonal-calving pasture-grazed dairy cows., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Recessive alleles represent genetic risk in populations that have undergone bottleneck events. We present a comprehensive framework for identification and validation of these genetic defects, including haplotype-based detection, variant selection from sequence data, and validation using knockout embryos. Holstein haplotype 2 (HH2), which causes embryonic death, was used to demonstrate the approach. Holstein haplotype 2 was identified using a deficiency-of-homozygotes approach and confirmed to negatively affect conception rate and stillbirths. Five carriers were present in a group of 183 sequenced Holstein bulls selected to maximize the coverage of unique haplotypes. Three variants concordant with haplotype calls were found in HH2: a high-priority frameshift mutation resulting, and 2 low-priority variants (1 synonymous variant, 1 premature stop codon). The frameshift in intraflagellar 80 (IFT80) was confirmed in a separate group of Holsteins from the 1000 Bull Genomes Project that shared no animals with the discovery set. IFT80-null embryos were generated by truncating the IFT80 transcript at exon 2 or 11 using a CRISPR-Cas9 system. Abattoir-derived oocytes were fertilized in vitro, and zygotes were injected at the one-cell stage either with a guide RNA and CAS9 mRNA complex (n = 100) or Cas9 mRNA (control, n = 100) before return to culture, and replicated 3 times. IFT80 is activated at the 8-cell stage, and IFT80-null embryos arrested at this stage of development, which is consistent with data from mouse hypomorphs and HH2 carrier-to-carrier matings. This frameshift in IFT80 on chromosome 1 at 107,172,615 bp (p.Leu381fs) disrupts WNT and hedgehog signaling, and is responsible for the death of homozygous embryos., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Endometrial epidermal growth factor (EGF) shows a cyclic change with two peaks on Days 2-4 and 13-14 during the estrous cycle. An altered (i.e., loss of the two peaks) profile has been linked to reduced fertility in repeat breeder cows. We previously demonstrated that a form of osteopontin (OPN), with a molecular weight of 29 kDa and found in bull seminal plasma (SP), normalized the EGF profile and restored fertility in repeat breeder cows. OPN has many molecular forms due to post-translational modifications and is abundant in bovine milk. The purpose of the present study was to investigate whether mOPN normalizes the endometrial EGF profile and restores fertility in repeat breeder dairy cows with an altered EGF profile. OPN was separated by one-step anion-exchange column chromatography from the whey of bovine milk. Purified mOPN was verified by Western blotting and peptide mass fingerprinting analyses. The OPN fraction showed three major protein bands of 61, 37 and 31 kDa (peptides I, II, and III, respectively) on SDS-PAGE. All three major bands were identified as OPNs by Western blotting and their tryptic peptide masses were matched at approximately 50, 40, and 10%, respectively, to the bovine OPN amino acid sequence by a peptide mass finger printing analysis. The three bands accounted for approximately 85% of the total protein content and 6-23 mg of OPN was obtained from 1 L of bovine milk. A lyophilized eluate containing 1.3 mg of mOPN (171 cows), 0.5 mL of frozen SP (62 cows), and PBS (84 cows) was infused at estrus into the vagina of repeat breeder cows with an altered EGF profile. Some of the cows treated with mOPN, SP, and PBS (46, 50, and 45 cows, respectively) were inseminated immediately before the infusion and then examined for pregnancy between Days 60 and 65. The rate at which mOPN to normalize the EGF profile (56.1%) was similar to that of SP (58.1%) and higher than that of PBS (23.8%) (P < 0.05). The conception rate after the infusion of mOPN (43.5%) was similar to that of SP (40.0%) and higher than that of PBS (22.2%) (P < 0.05). The present results indicate that the infusion of mOPN into the vagina is a treatment option for repeat breeder cows with an altered EGF profile. Further studies are needed to compare the capacity of the three OPN molecules in milk to normalize the EGF profile, together with their molecular characteristics due to post-translational modifications., (Copyright © 2022 Elsevier Inc. All rights reserved.)

The experimental objective was to examine the role of melatonin and its pathways in the maintenance of pregnancy in lactating dairy cows. Blood samples were collected at days 0, 16 and 32 after timed AI from cows (n = 200) in order to consider plasma melatonin concentrations and to conduct AOPP (advanced oxidation products of proteins) and TBARS (thiobarbituric acid reactive substances) tests. Luminal endometrial cells were collected at day 16 using a Cytobrush in all cows to determine mRNA expressions of melatonin receptor 1 ( MT1 ), mouse double minute 2 ( MDM2 ), MDM2 binding protein ( MTBP ), BCL2-associated X , apoptosis Regulator ( BAX ), p53 upregulated modulator of apoptosis (PUMA, gene symbol BBC3), mucin 1 ( MUC1 ) and leukemia inhibitory factor ( LIF ). Plasma concentrations of melatonin were significantly greater in pregnant cows diagnosed pregnant at day 16 who sustained pregnancy to day 32 compared to nonpregnant cows at day 16, or pregnant at day 16 and who lost embryos by days 32. Concentrations of AOPP and TBARS were greater in nonpregnant cows at day 16 or pregnant at day 16 and who lost embryos by days 32 compared to those diagnosed pregnant at day 16 and who sustained pregnancy to day 32. In pregnant cows, endometrial mRNA expressions of MDM2 , MTBP , MTR1 and LIF were higher compared to pregnant-embryo-loss cows ( p < 0.05). In contrast, mRNA expressions of BBC3 and MUC1 were greater at day 16 in pregnant-embryo-loss cows compared to pregnant cows ( p < 0.05). In conclusion, melatonin status is a modulator of embryo well-being and maintenance of pregnancy in lactating dairy cows.

Research Question: What is the impact of radiation exposure on oocyte quality and female fertility?, Design: Prepubertal mice underwent whole-body irradiation with a single dose (0.02, 0.1, 0.5, 2, 8 Gy) of gamma- or X-rays. Oocytes were quantified in irradiated (n = 36) and sham-treated (n = 8) mice. After a single exposure to 2 Gy, formation of DNA double-strand breaks (n = 10), activation of checkpoint kinase (Chk2) (n = 10) and dynamics of follicular growth (n = 18) were analysed. Fertility assessment was performed in adult irradiated mice and controls from the number of pups per mouse (n = 28) and the fetal abortion rate (n = 24). Ploidy of mature oocytes (n = 20) was analysed after CREST immunostaining, and uterine sections were examined., Results: Radiation exposure induced a massive loss of primordial follicles with LD 50 below 50 mGy for both gamma and X-rays. Growing follicles survived doses up to 8 Gy. This difference in radiosensitivity was not due to a different amount of radio-induced DNA damage, and Chk2 was activated in all oocytes. Exposure to a 2 Gy dose abolished the long-term fertility of females due to depletion of the ovarian reserve. Detailed analysis indicates that surviving oocytes were able to complete folliculogenesis and could be fertilized. This transient fertility allowed irradiated females to produce a single litter albeit with a high rate of fetal abortion (23%, P = 0.0096), related to altered ploidy in the surviving oocytes (25.5%, P = 0.0035)., Conclusions: The effects of radiation on surviving oocyte quality question natural conception as a first-line approach in cancer survivors. Together, the data emphasize the need for fertility preservation before radiation exposure and call for reassessment of the use of cryopreserved oocytes., (Copyright © 2021 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.)

This study aims to evaluate the association between reproductive disorders and the main serogroups of Leptospira spp. in dairy herds. Blood samples from 216 cows from nine herds were collected for a Microscopic Agglutination Test (MAT), and a reproductive follow-up with ultrasonography was carried out monthly for at least 12 months. A total of 75 (16.5%) cows were reactive, while seroreactivity to a herd level ranged from 16.0% to 52.8%. Out of the nine herds, in five (55.0%) Sejroe was found to be the predominant serogroup and in four (45.0%) it was Icterohaemorrhagiae. The major reproductive problems identified were embryonic death (ED) and abortions. All herds presented ED and abortion was observed in three (33.3%). Herds with high seroreactivity (>20%) had more reproductive problems. Those with a predominance of Sejroe were associated with ED (P < 0.05), while those with Icterohaemorrhagiae were associated with abortion. The results show that different leptospiral strains lead to different reproductive problems, as well as showing that the main problem - ED - is caused by leptospirosis, particularly for those determined by strains of the serogroup Sejroe., Competing Interests: Declaration of competing interest None., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Zika virus (ZIKV) is an arthropod-borne virus (arbovirus) and is primarily transmitted by Aedes species mosquitoes; however, ZIKV can also be sexually transmitted. During the initial epidemic and in places where ZIKV is now considered endemic, it is difficult to disentangle the risks and contributions of sexual versus vector-borne transmission to adverse pregnancy outcomes. To examine the potential impact of sexual transmission of ZIKV on pregnancy outcome, we challenged three rhesus macaques ( Macaca mulatta ) three times intravaginally with 1 x 10 7 PFU of a low passage, African lineage ZIKV isolate (ZIKV-DAK) in the first trimester (~30 days gestational age). Samples were collected from all animals initially on days 3 through 10 post challenge, followed by twice, and then once weekly sample collection; ultrasound examinations were performed every 3-4 days then weekly as pregnancies progressed. All three dams had ZIKV RNA detectable in plasma on day 3 post-ZIKV challenge. At approximately 45 days gestation (17-18 days post-challenge), two of the three dams were found with nonviable embryos by ultrasound. Viral RNA was detected in recovered tissues and at the maternal-fetal interface (MFI) in both cases. The remaining viable pregnancy proceeded to near term (~155 days gestational age) and ZIKV RNA was detected at the MFI but not in fetal tissues. These results suggest that sexual transmission of ZIKV may represent an underappreciated risk of pregnancy loss during early gestation., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Newman, Tarantal, Martinez, Simmons, Morgan, Zeng, Rosinski, Bliss, Bohm, Dudley, Aliota, Friedrich, Miller and O’Connor.)

Non-lactating multiparous NZW rabbit does (n = 227) were used in two experiments. In the 1st experiment (n = 87), does were i.m. injected with 0.1-ml saline/doe in day 0 (control, n = 29). Other does were injected with 25 IU equine chorionic gonadotrophin (eCG), followed by 0.2-ml gonadotrophin-releasing hormone (GnRH, n = 29) or 75 IU human chorionic gonadotrophin (hCG, n = 29) per doe 48 h later. After 60 h of day 0, does in all groups were artificially inseminated (AI). In the 2nd experiment, does (n = 140) were mated (AI) after synchronization of estrus/ovulation with 25 IU eCG, and 75 IU hCG 48 h later. On day 5 post-AI, does were injected with saline (control), 75 IU hCG, 0.2 ml GnRH, or 25 IU eCG per doe. Injection of eCG with GnRH or hCG pre-AI significantly increased corpora lutea number, ovulation rate, total number/doe and recovery rate of embryos, viable embryos, hatched blastocysts, in vivo reproductive parameters, and concentration of progesterone and progesterone/estradiol 17-β ratio. Injection of eCG on day 5 post-AI significantly improved large and total follicle number, and in vivo reproductive efficiency. The corpora lutea number and impantation sites were significantly increased in the hCG and eCG groups. Fetal loss rate significantly increased only in the GnRH group. Under high ambient temperature, administration of eCG with hCG or GnRH injection pre-AI could be synchronized estrus/ovulation for improving in vivo and in vitro embryo production. In addition, pregnancy outcomes could be enhanced in rabbit does induced to ovulation by a single eCG or hCG dose on day 5 post-AI.

The objective of this study was to evaluate the fertility of sexed semen compared with conventional semen with regard to the puberty and breeding ages of Holstein dairy heifers subjected to double Ovsynch protocol with fixed time of artificial insemination. A total of 468 Holstein heifers were divided into two groups. The first group was 122 dairy heifers inseminated via conventional semen, while the second group was 346 heifers inseminated with sexed semen. The puberty and breeding ages of heifers were determined from the farm records. Estrus was synchronized using the double Ovsynch protocol. Numbers were estimated for pregnancy at 40 and 60 days post insemination, embryonic loss, and abortion. The results revealed that the heifers inseminated with sexed semen had a significantly lower first-service pregnancy rate (51.45%) than those inseminated with conventional semen (61.47%). Heifers achieving puberty before 350 days old had a higher pregnancy rate. Embryonic losses and abortion rates did not differ between the two types of semen. Holstein heifers subjected to Ovsynch protocol with sexed semen had an acceptable first-insemination pregnancy rate. Even the applications of sexed semen reduce the reproductive fertility and pregnancy rate in Holstein heifers.

Epidermal growth factor (EGF) concentrations in the uterus show two peaks on days 2-4 and 13-14 during the estrous cycle in fertile cows. Loss of the two peaks has been linked to reduced fertility in repeat breeder cows. This study aimed to examine the effect of seminal plasma (SP) on normalizing endometrial EGF concentrations and restoring fertility in repeat breeder cows with low EGF concentrations on day 3. In study 1, we examined the effect of the deposition sites (the vagina and uterus) of SP on the endometrial EGF concentrations in repeat breeder cows. SP infusion into the vagina, but not uterus, on the first day of the estrus cycle (day 0) normalized the endometrial EGF concentrations (≥ 4.7 ng/g tissue weight) on day 3. In study 2, the effect of SP volume (0.5 and 10 ml of SP and 0.5 ml of SP diluted to 10 ml) on EGF concentrations was examined. All groups with SP infusion had increased EGF concentrations on day 3, and cows with 10 ml of SP and 0.5 ml of SP diluted to 10 ml showed the highest levels of EGF concentrations. In study 3, we examined the effect of SP infusion on fertility. SP infusion normalized two peaks of endometrial EGF concentrations in about 60% of repeat breeder cows and produced more pregnancies than the controls (44.4 vs. 19.4%). Therefore, we concluded that SP may contain an activity to normalize the EGF profile and restore fertility in repeat breeder cows with altered EGF profiles.

The reproductive performance of goats that received a GnRH analog (gonadorelin) fabricated with or without chitosan-sodium tripolyphosphate (TPP) nanoparticles on the day of estrus (day 0) was evaluated. The chitosan-TPP polymer was conjugated with gonadorelin using an ionic gelation method. Thirty-three multiparous Zaraiebi goats were synchronized for estrus with 2 intramuscular (im) injections of 125 μg prostaglandin F 2α 14 d apart. Goats showing signs of estrus were divided equally into 3 experimental groups and received a single im injection of 1 mL physiological saline (placebo; control), 50 μg/mL gonadorelin (GnRH), or 12.5 μg (quarter of GnRH dose)/mL chitosan-TPP-conjugated gonadorelin nanoparticles (NGnRH). Each goat underwent ultrasound imaging of their ovaries at day 0 and at day 10 after mating, and pregnancy was diagnosed 28 and 45 d after mating. The concentrations of estradiol (E 2 ) and progesterone (P 4 ) were determined at day 0 and at days 7, 14, 21, and 42 after mating. NGnRH size, polydispersity, and zeta potential were 93.91 nm, 0.302, and 11.6 mV, respectively. Chitosan-TPP nanoparticles showed 91.2% entrapment efficiency for GnRH. No differences in estrus rate, interval to estrus, or ovarian structure at day 0 were observed among the experimental groups, but the GnRH and NGnRH treatments significantly decreased the duration of estrus compared with the control. At day 10 after mating, both GnRH and NGnRH increased (P = 0.011) the number of corpora lutea compared with the control. Treatment with GnRH increased (P = 0.023) serum E 2 concentrations from day 7 to 42 after mating compared with NGnRH and control treatments. The highest (P = 0.043) serum P 4 concentration was observed in the GnRH group, followed by the NGnRH and control groups. The increase in serum P 4 concentration started earlier a on day 7 in the GnRH group but later on day 14 in the NGnRH group. Compared with the control, GnRH resulted in a higher (P = 0.041) P 4 -to-E 2 ratio, followed by NGnRH. Both gonadorelin treatments significantly increased the twinning rate, the number of embryos at days 28 and 42, and prolificacy and decreased pregnancy losses compared with the control. In conclusion, the administration of GnRH at the time of estrus improved the prolificacy of goats by increasing both the ovulation rate and the number of embryos. In addition, the nanoformulation developed in this study allowed a 75% reduction in the conventional dose of gonadorelin without affecting the fertility and prolificacy of goats, indicating the bioavailability of the reduced GnRH dose after conjugation with developed nanoformula., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Several tools exist to diagnose pregnancy in dairy cattle. However, substantial pregnancy loss occurs within the first 60 d of gestation in cattle, and these losses have a profound adverse economic impact on the dairy and beef cattle industries. Detecting these impending pregnancy losses could offer producers an opportunity to reduce costs associated with this source of reproductive inefficiency. Several of the pregnancy diagnostic tools currently available and new technologies are being examined for their ability to predict pregnancies at risk for failing in early pregnancy. This review provides a synopsis of work undertaken recently to predict pregnancy losses in cattle. Currently, opportunities to predict pregnancy loss include (1) using transrectal ultrasonography to detect loss of the fetal heartbeat, floating debris within the placental fluids, and reductions in fetal size; (2) observing reductions in circulating progesterone concentrations; (3) detecting reductions in concentrations of circulating placental products; namely, pregnancy-associated glycoproteins and microRNAs; and (4) detecting reductions in the early pregnancy-dependent increase in interferon-stimulatory gene expression in peripheral blood leukocytes. An achievable goal may be to identify markers of embryo mortality so that researchers and clinicians can focus their efforts on developing intervention strategies for cows identified to be at risk for pregnancy failure., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Background: In vitro embryo production (IVEP) is increasingly popular but data assessing the outcome of transferred embryos are scarce., Objectives: To determine the likelihood of pregnancy and embryonic loss after transfer of frozen-thawed IVP embryos and identify factors influencing success., Study Design: Retrospective clinical study., Methods: Blastocysts (n = 261) were produced from immature oocytes of Warmblood mares (n = 116) by Intracytoplasmic Sperm Injection (ICSI) and in vitro culture, and cryopreserved. Thawed IVP embryos were transferred into recipient mares on day 4, 5 or 6 after ovulation. The influence of donor mare (age, reproductive history), recipient mare (age, reproductive status, management; in-house vs. outpatient, day post-ovulation), embryo (interval from ICSI to blastocyst formation) and management factors (season when ovum pickup was performed, year and method of transfer) on likelihood of pregnancy and embryonic loss was examined, and the developmental stage of the IVP embryo at the time of transfer was estimated., Results: The percentage of mares pregnant 7-10, 23 and 37 days after transfer was 56% (147/261), 49% (129/261), and 48% (124/261), respectively. Development of IVP embryos after transfer equated to day 5 or 6 in vivo embryos. With the exception of year of transfer, none of the factors had an impact on the likelihood of pregnancy or embryonic loss. Nevertheless, the likelihood of pregnancy tended to be lower for IVP embryos from infertile mares or when embryos were transferred into recipient mares on day 6 after ovulation rather than on day 4 or 5. Finally, the diameter of the embryonic vesicle 7 days post transfer was lower for pregnancies that were lost compared to those that were maintained., Main Limitations: Small sample size in some of the donor and recipient mare categories., Conclusions: Cryopreserved IVP embryos should be transferred into recipient mares on day 4 or 5 after ovulation and a slower rate of post transfer vesicle expansion indicates a higher risk of subsequent embryonic loss The Summary is available in Portuguese - see Supporting Information., (© 2018 EVJ Ltd.)

The feeding of diets with greater energy content than that needed for body maintenance following mating is believed to reduce embryonic survival in pigs. In swine operations, therefore, feed intake is often restricted during the first and second week of pregnancy to reduce embryo mortality. There is thought to be a relationship between feeding diets that result in energy intake that is greater than that needed for body maintenance and embryonic death. This relationship is associated with lesser than typical progesterone (P4) concentrations when feeding diets with greater energy content due to increased hepatic clearance. There is no consensus, however, as to whether feeding should be restricted during early pregnancy to avert this possible detrimental effect. Thus, the aim of this systematic review is to assess the effect in sows and gilts of feeding diets with different energy contents post-mating on embryonic survival, evaluating when possible, the relationship of a greater energy intake and P4 concentrations on embryonic survival. An electronic search was conducted of the PubMed, Science Direct, Scopus, Web of science, and Scielo databases during June 2018. A total of 109 articles were retrieved, and of these, only 16 articles were selected after applying the selection criteria. There was no negative effect of a greater feed intake than that needed for body maintenance after breeding in 75% of the experiments. Results from 35% of the experiments indicated feeding early pregnant sows a diet with greater energy content than that needed for body maintenance resulted in augmented embryonic death. In 66.7% of the experiments, in which there was assessment of P4 concentration, there was no negative effect of feeding after farrowing a diet with greater energy than that needed for body maintenance. In conclusion, it appears that restricted feed intake in early pregnancy is no longer relevant when there are modern prolific dam lines utilized in swine production enterprises because dietary energy of as great as 54 MJ ME/day had no detrimental effect on embryo survival., (Copyright © 2019 Elsevier B.V. All rights reserved.)

High reproductive performance is an element factor for profitability in dairy herds; although, pregnancy loss of non-infectious nature during early and mid-to-late embryonic period is increasing in dairy cattle. Based on that, the present study aimed to determine the incidence of pregnancy loss before and after 60 days of insemination, and to identify some non-infectious causes that could enhance it. The herd was composed by 600 crossbred dairy cows and those with a corpus luteum (CL) were treated with prostaglandinF2α, then inseminated, on the other hand, those without a CL were submitted to a timed artificial insemination protocol (TAI). Pregnancy losses rates were analyzed by logistic regression by SAS, and differences were considered significant when P < 0.05. The overall pregnancy loss and mid-to-late pregnancy loss were not affected by animal category (P > 0.05); although, early pregnancy loss was higher in cows than in heifers (11.90 vs. 3.39%). The early pregnancy loss was higher in those cows that calved on spring/summer when compared to those calved on fall/winter (9.22% vs. 16.11%), moreover, those inseminated during spring/summer tended to have higher early pregnancy loss when compared to those inseminated on fall/winter (13.35% vs. 8.57%). In conclusion, when some of non-infectious causes were evaluated, it was observed that cows that calved on spring/summer had higher pregnancy loss. At this point, strategies should be developing to minimize pregnancy loss in dairy herds, as it could be considered an important reproductive problem.

The goal of this study was to evaluate the effect of long-acting progesterone supplementation on pregnancy rates and gestational losses in Nelore females (Bos taurus indicus) submitted to Timed Artificial Insemination (TAI). Ovulations of multiparous (n = 534), primiparous (n = 117) and nulliparous (n = 81) Nelore females were synchronized for TAI and the day of insemination was considered D0. After TAI, the animals were allocated into three groups. Two groups received 150 mg of long acting injectable progesterone (P4) in a single dose, 5 (D = 5) or 11 (D = 11) days after TAI, resulting in the following treatments: 1) Group P4 - D5 (n = 235); 2) Group P4 - D11 (n = 245); and 3) Control Group (n = 252). The animals in the control group did not receive any supplementation. Pregnancy was checked by ultrasonography, 30 days after the TAI. The gestational losses were evaluated at two different periods: 1) from 30 to 60 days, and 2) from 60 to 90 days. The pregnancy rate and the gestational losses as a function of the treatments were tested by the non-parametric binomial test with 5% significance level. The pregnancy rates were different between the groups P4 - D5 and Control (47 and 39%, respectively) (P < 0.05), but no difference was observed in relation to the P4 - D11 group. There was a difference in gestational losses from 30 to 60 days and total losses between the groups treated with P4 on the fifth day and eleventh day after TAI in relation to the control group (P = 0.0173; P = 0.0454), but no difference was observed between the treatments in relation to the losses between 60 and 90 days (P ≥ 0.0578). Among the animals that were cycling at the beginning of the protocol, the groups supplemented with progesterone at day 5 or 11 after insemination had gestational losses significantly lower (P = 0.04) than the control group (P4 - D5 = 2%; P4 - D11 = 2%, control = 14%). The animals in anestrous that received supplementation 5 days after TAI had lower gestational losses (P = 0.049) compared with the other groups (P4 - D5 = 3%, P4 - D11 = 7% Control = 10%). Under the conditions of this study, the use of long-acting injectable progesterone 5 days after TAI in Nelore females increased the pregnancy rates, but when supplemented 11 days after TAI, it did not have the same effect. Also, when used 5 days after TAI, it significantly decreased gestational losses on both anestrous and cycling animals. Therefore, it may be an interesting alternative to increase reproductive and productive efficiencies., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Background: Pregnancy loss is a major source of infertility in dairy cows. Despite a fertilization rate after insemination (AI) of approximately 90%, calving rates are 30%-50%, indicating the occurrence of extensive embryonic and foetal losses. The aim of this study was to establish the extent and pattern of embryonic and foetal loss in Swedish Red (SR) and Swedish Holstein (SH) dairy cows, as well as, the relationship to oestrus intensity (OI) and progesterone (P4) concentration. In total, 2130 AIs and 16,176 milk P4 samples from 359 SR and 212 SH dairy cows were included in the study. Pregnancy losses were estimated using data from P4 values combined with AI information and calving data., Results: Total pregnancy loss from AI to the day of calving was 65%. Early embryonic loss, late embryonic loss and foetal loss were estimated to be 29, 14 and 13%, respectively. There is strong evidence in the literature that P4 concentrations at different time points are associated with pregnancy loss. In the present study, cows with pregnancy losses had significantly higher P4 levels at the day of AI and significantly lower P4 concentration at days 10, 21 and 30 after AI compared to pregnant cows. Swedish Red cows had significantly lower total pregnancy losses compared to SH cows (62% and 68% respectively, P = 0.017). Early embryonic loss was 6.7% points lower for cows inseminated at a stronger OI (OI = 3) compared to at a weaker OI (OI = 2, P = 0.006). Cows inseminated at ovulation number ≥ 5 had significantly lower early pregnancy losses compared to cows inseminated at first or second ovulation (11.5 and 8% points, respectively, P < 0.05). With an increase of one SD of milk (448 kg ECM) during the first 60 days in milk, early embryonic loss increased by 4.7% points (P = 0.006)., Conclusions: It is important to increase the number of cows calving per insemination by reducing embryo/foetal loss. This outcome can be achieved by management and breeding for optimal P4 levels at critical time points, and by considering oestrus expression in the breeding programmes to facilitate the correct timing of insemination.

This study aimed to investigate the effects of a single administration of one of three different gonadotropins on Day 7 post-insemination on ovarian activity, progesterone (P 4 ) concentration and pregnancy outcomes of rabbit does. Multiparous, non-lactating, V-line does were artificially inseminated after synchronization and ovulation induction with equine chorionic gonadotropin (eCG; 25 IU im) and gonadotropin releasing hormone (GnRH; 0.8  μg buserelin im) 48 h later. On Day 7 post-inseminarion, does were randomly allocated into four groups (n = 40/group). Does of each group were intramuscularly injected with a single dose of one of physiological saline (placebo; control), GnRH (0.8  μg buserelin), human chorionic gonadotropin (hCG; 25 IU) or eCG (25 IU). Concentration of serum P 4 was determined on Days 6, 9, 11 and 18 post-insemination. On Day 14 post-insemination, the ovaries and reproductive tracts of pregnant does were removed and weighed. Also, numbers of visible follicles, hemorrhagic follicles, corpora lutea of pregnancy (pCLs), new CLs (nCLs; formed after Day 7 post-insemination) and implantation sites were recorded. Conception rate, parturition rate, abortion rate, litter size/weight and litter viability were recorded. The highest (P < 0.05) reproductive tract and ovary weights were for eCG. The highest (P < 0.05) number of visible ovarian follicles was for eCG, whereas the lowest (P < 0.05) was for GnRH. Treatment with eCG increased (P < 0.05) numbers of pCLs and total implantation sites compared to the other groups. Treatment with GnRH or hCG increased (P < 0.05) number of nCLs compared to control and eCG. The highest rate of fetal loss was in does treated with GnRH. The concentration of serum P 4 decreased (P < 0.05) following the treatment with GnRH and continued low until Day 18. However, it remained in line for control, hCG and eCG groups up to Day 11, then decreased (P < 0.05) for control and hCG on Day 18, being lower for hCG than control, while continued to increase for eCG up to Day 18. Compared to control, treatment with eCG improved (P < 0.05) conception and parturition rates by 24 and 22%; respectively, while GnRH and hCG treatments decreased (P < 0.05) them by 57 and 47.6%; respectively. Litter size and litter weight at birth were improved by eCG, but were adversely affectd by GnRH and hCG. In conclusion, a single administration of eCG 7 Days post-insemination could be recommended for improving pregnancy outcomes in rabbits., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Defining factors associated with embryonic and/or fetal losses will be helpful in overcoming such problem, either by adjusting conditions or applying therapeutic approaches to high-risk cows. The objective of this study was to investigate the association between a number of maternal and non-maternal factors and the risk of late embryonic (LED) and early fetal death (EFD) in dairy herds. Additionally, we investigated the effect of treating pregnant cows either with GnRH on day 26 post-insemination, or GnRH on day 26 plus CIDR insertion between days 26 and 33 post-insemination, on the risk of LED/EFD. From 3826 pregnancies, diagnosed at day 30 post-insemination, 851 cows lost the pregnancy by day 70 post-insemination. A mixed-effects logistic model was constructed to assess the effect of cow breed, calving difficulty, postpartum problems, lactation number, days in milk, insemination number, actual 305-day milk production, temperature humidity index (THI) at insemination, estrus synchronization protocols, and other factors, on the risk of LED/EFD. Our findings indicated that Holstein X Brown Swiss crossbreed cows had a lower risk for LED/EFD than Holstein cows (P < 0.05). Cows that had postpartum problems, were inseminated for the first time, produced more milk, or were inseminated at THI ≥75, recorded higher risks of LED/EFD (P < 0.05). Calving difficulty, lactation number, and synchronization protocols were not found to be associated with LED/EFD. Moreover, treatment of the pregnant cows with GnRH on day 26 post-insemination plus CIDR insertion between days 26 and 33 post-insemination decreased the risk of LED/EFD. In conclusion, cows that had postpartum problems, were inseminated early postpartum, produced higher milk, and/or were inseminated at high THI, were under higher risk of LED/EFD. Treating such cows with GnRH on day 26 plus CIDR insertion between days 26 and 33 may decrease the possibility of the LED/EFD., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Early pregnancy loss occurs in 6-10% of equine pregnancies making it the main cause of reproductive wastage. Despite this, reasons for the losses are known in only 16% of cases. Lack of viable conceptus material has inhibited investigations of many potential genetic and pathological causes. We present a method for isolating and culturing placental cells from failed early equine pregnancies. Trophoblast cells from 18/30 (60%) failed equine pregnancies of gestational ages 14-65 days were successfully cultured in three different media, with the greatest growth achieved for cells cultured in AmnioChrome™ Plus. Genomic DNA of a suitable quality for molecular assays was also isolated from 29/30 of these cases. This method will enable future investigations determining pathologies causing EPL., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

An autosomal recessive genetic defect termed JH1 has been associated with early embryonic loss in the Jersey cattle breed. The genetic basis has been identified as a cytosine to thymine mutation in the CWC15 gene that changes an amino acid from arginine to a stop code. To screen for JH1 carriers in an imported Jersey population in China, a method based on a polymerase chain reaction amplification followed by a restriction fragment length polymorphism assay (PCR-RFLP) was developed for the accurate diagnosis of the JH1 allele. A total of 449 randomly chosen cows were examined with the PCR-RFLP assay, and 31 were identified as JH1 carriers, corresponding to a carrier frequency of 6.9%. The PCR-RFLP method was validated by DNA sequencing of 8 positive and 13 negative samples, with all 21 samples giving the expected DNA sequence. In addition, 3 negative and 3 positive samples were confirmed by a commercial microarray-based single nucleotide polymorphism assay. Finally, samples from 9 bulls in the United States of known status were correctly identified as carriers (5 bulls) or noncarriers (4 bulls). As the JH1 defect has most likely spread worldwide, implementing routine screening is necessary to avoid the risk of carrier-to-carrier matings and to gradually eradicate the deleterious gene., (© 2015 The Author(s).)