Effects of carprofen and/or CIDR administration on pregnancies per artificial insemination around pregnancy recognition in lactating dairy cows

Effect of carprofen and/or CIDR on pregnancies per AI (P/AI) 14 days after artificial insemination (AI) in lactating cows were investigated. Following detection of corpus luteum (CL) by ultrasonography (USG) 14 days after AI, cows (n=853) were randomly allocated to CARP (carprofen, 1.4 mg/kg; Rimadyl®XL), CIDR (progesterone, 1.38g, for seven days), CARP+CIDR and CONT (control) groups. CL was re-determined with USG 21 days after AI to monitored the maintenance of CL. Rates of maintenance of CL did not differed among CARP (79.6% [168/211]), CIDR (86.0% [196/228]), CARP+CIDR (80.0% [172/215]) and CONT (74.9% [149/199]) groups; however, the interaction effect of treatment by body condition score (BCS) at AI (P<0.05) were existed. In this matter, among cows with BCS≤2.5, chances of maintenance of CL was lower in CONT (70.7%, [111/157]) group compared to those in CARP (81.7%, [125/153]), CIDR (83.6%, [153/183]) and CARP+CIDR (80.1%, [129/161]) groups. Whereas, among cows with BCS>2.5, chances of maintenance of CL were lower in CARP (74.1%, [43/58]) and CARP+CIDR (79.6%, [43/54]) groups compared to those in CONT (90.5%, [38/42]) and CIDR (95.6%, [43/45]) groups. For the P/AI at 28-32 days after AI, there was no difference among CARP (48.8% [103/211]), CIDR (50.9 [116/228]), CARP+CIDR (47.4% [102/215]) and CONT (44.7% [89/199]) groups. Pregnancy losses between 28-32 and 55-60 days did not differ among CARP (3.9%, [4/103]), CIDR (4.3%, [5/116]), CARP+CIDR (5.9%, [6/102]) and CONT (6.7%, [6/89]) groups. However, there was a significant (P<0.05) interaction effect of treatment by the number of services on pregnancy losses. In this regard, pregnancy losses were higher in cows inseminated thrice and four or more times in CONT (11.8% and 16.7%) group compared to those in CARP (0% and 4.8%), CIDR (0% and 6.9%), CARP+CIDR (0% and 11.1%); respectively. Consequently, no effects of carprofen or CIDR around pregnancy recognition on P/AI were observed despite a higher maintenance rate of CL in lactating cattle. Furthermore, fewer pregnancy losses in cows following three or more services could indicate the beneficial carry-over effects of carprofen and/or CIDR administration around maternal recognition of pregnancy.

The Importance of Interferon-Tau in the Diagnosis of Pregnancy

Several decades of improving dairy cattle towards unilateral utilization of dairy cattle led to enormous progress in the field of milk yield; however, it resulted in a number of unfavorable features, such as reproductive disorders, increased calf mortality, and reduced health. Most cases of embryo loss and/or lost pregnancies occur during the first four to five weeks of gestation; accurate detection for pregnancy during this period is likely to contribute to an improvement in gestation rates. A specific protein, interferon-tau (IFNT), stimulates interferon-stimulated genes (ISGs), and their expression increases during gestation within 21 days after insemination. In bovines, the early conceptus undergoes a phase of rapid growth and elongation before implantation, the latter occurring 2–3 weeks after fertilization. IFNT acts mainly in the endometrium of the luminal epithelium. It is a new type I interferon that regulates several genes encoding uterine-derived factors. They are crucial in the processes of preparing the uterus for placenta attachment, modifying the uterine immune system, and regulating early fetal development. Because IFNT is expressed and induces ISGs in the endometrium during pregnancy recognition, it was reasoned that surrogate markers for pregnancy or IFNT might be present in the blood and provide an indicator of pregnancy status in cattle.

Uterine Notch2 facilitates pregnancy recognition and corpus luteum maintenance via upregulating decidual Prl8a2

The maternal recognition of pregnancy is a necessary prerequisite for gestation maintenance through prolonging the corpus luteum lifespan and ensuring progesterone production. In addition to pituitary prolactin and placental lactogens, decidual derived prolactin family members have been presumed to possess luteotropic effect. However, there was a lack of convincing evidence to support this hypothesis. Here, we unveiled an essential role of uterine Notch2 in pregnancy recognition and corpus luteum maintenance. Uterine-specific deletion of Notch2 did not affect female fertility. Nevertheless, the expression of decidual Prl8a2, a member of the prolactin family, was downregulated due to Notch2 ablation. Subsequently, we interrupted pituitary prolactin function to determine the luteotropic role of the decidua by employing the lipopolysaccharide-induced prolactin resistance model, or blocking the prolactin signaling by prolactin receptor-Fc fusion protein, or repressing pituitary prolactin release by dopamine receptor agonist bromocriptine, and found that Notch2-deficient females were more sensitive to these stresses and ended up in pregnancy loss resulting from abnormal corpus luteum function and insufficient serum progesterone level. Overexpression of Prl8a2 in Notch2 knockout mice rescued lipopolysaccharide-induced abortion, highlighting its luteotropic function. Further investigation adopting Rbpj knockout and DNMAML overexpression mouse models along with chromatin immunoprecipitation assay and luciferase analysis confirmed that Prl8a2 was regulated by the canonical Notch signaling. Collectively, our findings demonstrated that decidual prolactin members, under the control of uterine Notch signaling, assisted pituitary prolactin to sustain corpus luteum function and serum progesterone level during post-implantation phase, which was conducive to pregnancy recognition and maintenance.

Comparison of the Ability of High and Low Virulence Strains of Non-cytopathic Bovine Viral Diarrhea Virus-1 to Modulate Expression of Interferon Tau Stimulated Genes in Bovine Endometrium

Bovine Viral Diarrhea virus (BVDV) is a pestivirus with a single-stranded, positive sense RNA genome. It is endemic in many cattle populations, causing major economic losses in part due to reduced fertility. BVDV exhibits great genetic diversity and is classified as type 1 or 2 (BVDV-1, BVDV-2) with either non-cytopathogenic (ncp) or cytopathogenic (cp) biotypes. Differing strains of ncpBVDV differ in virulence, affecting clinical outcome. BVDV replicates in the reproductive tract, affecting host immunity and embryo survival. This study used an in vitro model of primary bovine endometrial cell cultures to compare the effects of two BVDV ncp type 1a strains of differing virulence (termed HO and KY) on endometrial transcription of candidate interferon stimulated genes (ISG) using qPCR. Half the cultures were stimulated with interferon tau (IFNT, the conceptus produced pregnancy recognition factor) in the presence or absence of viral infection. Cultures were replicated on cells from 10 BVDV-free cows. IFNT treatment stimulated transcription of 10 candidate ISGs, whereas both ncpBVDV-1 strains alone inhibited transcription of 8/10 ISGs. In combined BVDV-1+IFNT cultures, the stimulatory effect of IFNT on expression of GBP4, ISG15, HERC5, RSAD2, IFIH1, IFIT3, and MX1 was significantly inhibited by HO, but only ISG15, RSAD2, IFI27, and IFIT3 were decreased by KY. Inhibition by HO was generally greater. The IFNT-induced expression of TRIM56 was, however, increased by HO. These data show that HO, the more virulent ncpBVDV-1 strain, has a greater capacity to inhibit key antiviral pathways. These differences need confirmation at the protein level but may influence immune tolerance of the host. They could also reduce fertility by increasing uterine susceptibility to bacterial infection and disrupting IFNT-mediated pregnancy recognition.

Conceptus metabolomic profiling reveals stage-specific phenotypes leading up to pregnancy recognition in cattle

Reproductive efficiency in livestock is a major driver of sustainable food production. The poorly-understood process of ruminant conceptus elongation (a) prerequisites maternal pregnancy recognition, (b) is essential to successful pregnancy establishment, and (c) coincides with a period of significant conceptus mortality. Conceptuses at five key developmental stages between Days 8–16 were recovered and cultured in vitro for 6 h prior to conditioned media analysis by untargeted ultrahigh-performance liquid chromatography tandem mass spectroscopy. This global temporal biochemical interrogation of the ex situ bovine conceptus unearths two antithetical stage-specific metabolic phenotypes during tubular (metabolically retentive) vs. filamentous (secretory) development. Moreover, the retentive conceptus phenotype on Day 14 coincides with an established period of elevated metabolic density in the uterine fluid of heifers with high systemic progesterone—a model of accelerated conceptus elongation. These data, combined, suggest a metabolic mechanism underpinning conceptus elongation, thereby enhancing our understanding of the biochemical reciprocity of maternal-conceptus communication, prior to maternal pregnancy recognition.

Cross-Country Abortion Travel to England and Wales: Results From a Cross-Sectional Survey Exploring Women’s Experiences Crossing Borders to Obtain Care.

Background: The laws governing abortion access vary across Europe. Even in countries with relatively liberal laws, numerous barriers to abortion access exist. In response to these barriers, evidence suggests that people living in countries with both restrictive and liberal laws travel outside of their home country for abortion care. England and Wales are common destinations for those who travel to seek abortions, but little is known about the motivations and experiences of those who undertake cross-country travel to England or Wales to obtain care. This paper aims to describe the abortion seeking and travel experiences of women and pregnant people who traveled to England and Wales for an abortion between 2017 and2019. Methods: We recruited 97 participants who had travelled cross-country from both liberal and restrictive contexts to seek abortion care at three participating BPAS clinics in England and Wales. Participants completed an electronic survey about their reproductive histories, abortion decision-making, experiences seeking abortion care and traveling. We conducted a descriptive analysis, and include comparisons between participants who traveled from liberal and restrictive contexts. Results: Over a third of participants considered abortion four weeks or more before presenting for care at BPAS, and around two-thirds sought abortion services in their home country before traveling. The majority of participants indicated that they would have preferred to have obtained an abortion earlier and cited reasons including scheduling issues, a dearth of local services, delayed pregnancy recognition, and financial difficulties as causing their delay. About seventy percent of participants reported travel costs between €101-1000 and 75% of participants reported that the cost of the abortion procedure exceeded €500. About half (53%) of participants indicated that, overall, their travel was very or somewhat difficult. Conclusions: This analysis documents the burdens associated with cross-country travel for abortion and provides insight into the factors that compel people to travel. Our findings highlight the need for expanded access to abortion care throughout Europe via the removal of legal impediments and other social or procedural barriers. Removing barriers would eliminate the need for cumbersome abortion travel, and ensure that all people can obtain necessary, high-quality healthcare in their own communities. Plain English summary: In Europe, women who live in countries where abortion is severely restricted or illegal altogether lack access to abortion care entirely, but even women who live in countries with more liberal laws face barriers due to gestational age limits, waiting periods, and a lack of trained and willing providers. Existing evidence suggests that restrictions and barriers compel women from both countries with restrictive laws as well as those from countries with liberal laws to travel outside of their home country for abortion services. England and Wales are common destinations for people traveling within Europe to obtain abortion services. However, little is known about the experiences of people who travel cross-country to England or Wales for abortions. We surveyed individuals who had traveled from another country to seek abortion services in England or Wales. Our analysis documents that many participants contemplated getting an abortion and sought care in their home countries before traveling. Likewise, many participants indicated that they would have preferred to have obtained an abortion earlier in their pregnancy, and referenced scheduling issues, a dearth of local services, delayed pregnancy recognition, and financial difficulties as causing their delay. A majority of participants indicated that covering the costs of their abortion, and the costs of travel was difficult, and that the travel experience in its entirety was difficult. Our findings document the reasons for, and burdens associated with abortion travel and highlight the need to expand access to abortion across Europe via the elimination of all legal restrictions and impediments.

Profile of Mothers of Children with Fetal Alcohol Spectrum Disorder: A Population-Based Study in Canada

Objective: To compare the characteristics of mothers of children with Fetal Alcohol Spectrum Disorder (FASD) with mothers of typically developing control children. Methods: The study utilized a cross-sectional, observational design, using active case ascertainment. Biological mothers were interviewed using a standardized retrospective questionnaire to collect data on demographics, living environment, pregnancy history, nutrition, alcohol and other drug use prior to and following pregnancy recognition. Results: A total of 173 mothers were interviewed. Of these, 19 had a child who was diagnosed with FASD, five had a child who had received a deferred FASD diagnosis, and 37 had children who were selected into the control group as typically developing children. The remaining 112 mothers had children who did not meet diagnostic criteria for FASD. The mothers of children with FASD did not differ significantly from mothers of the control group children with respect to age, ethnicity, marital status, and employment status at the time of pregnancy. However, mothers of children with FASD had lower levels of education (p < 0.01) and were more likely to have received financial support (p < 0.05) at the time of pregnancy, to have smoked tobacco (p < 0.001), and to have used marijuana or hashish (p < 0.01) prior to pregnancy recognition, compared with mothers of control children. All mothers of children with FASD reported alcohol consumption prior to pregnancy recognition; however, only 10.5% reported alcohol consumption following pregnancy recognition. None of the mothers interviewed reported any drug use following pregnancy recognition. Conclusions: Population-based preventive interventions, including repeated screening, monitoring, and education regarding the effects of alcohol use, as well as other substances, before and during pregnancy, are needed to eliminate risk for FASD and other negative consequences on child and maternal health.

Protein Synthesis by Day 16 Bovine Conceptuses during the Time of Maternal Recognition of Pregnancy

Interferon Tau (IFNT), the conceptus-derived pregnancy recognition signal in cattle, significantly modifies the transcriptome of the endometrium. However, the endometrium also responds to IFNT-independent conceptus-derived products. The aim of this study was to determine what proteins are produced by the bovine conceptus that may facilitate the pregnancy recognition process in cattle. We analysed by mass spectrometry the proteins present in conceptus-conditioned media (CCM) after 6 h culture of Day 16 bovine conceptuses (n = 8) in SILAC media (arginine- and lysine-depleted media supplemented with heavy isotopes) and the protein content of extracellular vesicles (EVs) isolated from uterine luminal fluid (ULF) of Day 16 pregnant (n = 7) and cyclic (n = 6) cross-bred heifers on day 16. In total, 11,122 proteins were identified in the CCM. Of these, 5.95% (662) had peptides with heavy labelled amino acids, i.e., de novo synthesised by the conceptuses. None of these proteins were detected in the EVs isolated from ULF. Pregnancy-associated glycoprotein 11, Trophoblast Kunitz domain protein 1 and DExD-Box Helicase 39A were de novo produced and present in the CCM from all conceptuses and in previously published CCM data following 6 and 24 h. A total of 463 proteins were present in the CCM from all the conceptuses in the present study, and after 6 and 24 h culture in a previous study, while expression of their transcripts was not detected in endometrium indicating that they are likely conceptus-derived. Of the proteins present in the EVs, 67 were uniquely identified in ULF from pregnant heifers; 35 of these had been previously reported in CCM from Day 16 conceptuses. This study has narrowed a set of conceptus-derived proteins that may be involved in EV-mediated IFNT-independent embryo–maternal communication during pregnancy recognition in cattle.

Alcohol drinking in pregnant women from an Argentinian city

Alcohol drinking during pregnancy might lead to detrimental consequences to the mother and fetus, being Fetal Alcohol Syndrome (FAS) the most severe. In many Latin-American countries, there is a paucity of information about prenatal exposure to alcohol. Aim. To estimate alcohol drinking in pregnant women and describe it concerning sociodemographic characteristics. Method. Data were collected from a random sample of women attending the public health care system (N=852) in 2016. Alcohol drinking, including binge drinking (BD), was evaluated before and after pregnancy recognition. Descriptive and bivariate analyses were performed. Results. While pregnant, two out of ten women reported drinking. Moreover, 2% engaged in BD and 3% presented indicators of alcohol dependence. BD before pregnancy was related to drinking during pregnancy and frequency, quantity, and BD before pregnancy with BD during pregnancy. Conclusions. Alcohol consumption in pregnant women was higher than the estimated for most Latin-American countries, and of the considered characteristics, was almost exclusively related to consumption patterns before pregnancy.

Expression of isg15 and conjugating enzyme during peri-implanation period in sheep

Pregnancy recognition in ruminant affects numerous genes regulation. Interferon-stimulatory gene (ISG15), an ubiquitin-like protein that mediates theconjugation of different proteins through its ISGylation enzymes UBE1L and UBCH8, is also differentially expressed during early pregnancy. The purpose of this study was to investigate the role of ISG-15 in the establishment of pregnancy and conceptus elongation during early post conception periods and to ascertain the presence of ISGylation enzymes UBE1L and UBCH8. Therefore, sheep were synchronized through cloprostenol sodium and gonadotrophin releasing hormone-1 and serviced by rams. The blood was collected on the post-mating days 6, 11, 12, 13, 14, 16, 17, 21, 23 and 25. The ISG-15, UBE1L and UBCH8 primers were used to amplify the corresponding transcriptomic region using PCR. Recovery rate of each transcriptomic fragment was compared with the housekeeping gene GAPDH. ISG-15 expression was higher on day 12, contrary to UBE1L were higher on day 6 and UBCH8 on day 21. Furthermore, the ISG-15 is ubiquitin-like protein, mediates UBE1L and UBCH8 enzymes to guard the conceptus against viral pathogenicity during early pregnancy.