Single-cell transcriptional landscapes of bovine peri-implantation development.

Supporting healthy pregnancy outcomes requires a comprehensive understanding of the molecular and cellular programs of peri-implantation development, when most pregnancy failure occurs. Here, we present single-cell transcriptomes of bovine peri-implantation embryo development at day 12, 14, 16, and 18 post-fertilization. We defined the cellular composition and gene expression of embryonic disc, hypoblast, and trophoblast lineages in bovine peri-implantation embryos, and identified markers and pathway signaling that represent distinct stages of bovine peri-implantation lineages; the expression of selected markers was validated in peri-implantation embryos. Using detailed time-course transcriptomic analyses, we revealed a previously unrecognized primitive trophoblast cell lineage. We also characterized conserved and divergence peri-implantation lineage programs between bovine and other mammalian species. Finally, we established cell-cell communication signaling underlies embryonic and extraembryonic cell interaction to ensure proper early development. These data provide foundational information to discover essential biological signaling underpinning bovine peri-implantation development.

Maternal restricted- and over- feeding during gestation perturb offspring sperm epigenome in sheep.

In brief: Inadequate maternal nutrition during gestation can have immediate and lifelong effects on offspring. This study shows that maternal restricted - and over- nutrition during gestation do not affect semen characteristics in F1 male offspring but alters offspring sperm sncRNA profiles and DNA methylome in sheep. Abstract: There is a growing body of evidence that inadequate maternal nutrition during gestation can have immediate and lifelong effects on offspring. However, little is known about the effects of maternal nutrition during gestation on male offspring reproduction. Here, using a sheep model of maternal restricted - and over - nutrition (60 or 140% of the National Research Council requirements) during gestation, we found that maternal restricted - and over - nutrition do not affect semen characteristics (i.e. volume, sperm concentration, pH, sperm motility, sperm morphology) or scrotal circumference in male F1 offspring. However, using small RNA sequencing analysis, we demonstrated that both restricted - and over - nutrition during gestation induced marked changes in composition and expression of sperm small noncoding RNAs (sncRNAs) subpopulations including in male F1 offspring. Whole-genome bisulfite sequencing analysis further identified specific genomic loci where poor maternal nutrition resulted in alterations in DNA methylation. These findings indicate that maternal restricted - and over - nutrition during gestation induce epigenetic modifications in sperm of F1 offspring sperm in sheep, which may contribute to environmentally influenced phenotypes in ruminants.

Single-cell transcriptional landscapes of bovine peri-implantation development.

Supporting healthy pregnancy outcomes requires a comprehensive understanding of the cellular hierarchy and underlying molecular mechanisms during peri-implantation development. Here, we present a single-cell transcriptome-wide view of the bovine peri-implantation embryo development at day 12, 14, 16 and 18, when most of the pregnancy failure occurs in cattle. We defined the development and dynamic progression of cellular composition and gene expression of embryonic disc, hypoblast, and trophoblast lineages during bovine peri-implantation development. Notably, the comprehensive transcriptomic mapping of trophoblast development revealed a previously unrecognized primitive trophoblast cell lineage that is responsible for pregnancy maintenance in bovine prior to the time when binucleate cells emerge. We analyzed novel markers for the cell lineage development during bovine early development. We also identified cell-cell communication signaling underling embryonic and extraembryonic cell interaction to ensure proper early development. Collectively, our work provides foundational information to discover essential biological pathways underpinning bovine peri-implantation development and the molecular causes of the early pregnancy failure during this critical period. Significance Statement: Peri-implantation development is essential for successful reproduction in mammalian species, and cattle have a unique process of elongation that proceeds for two weeks prior to implantation and represents a period when many pregnancies fail. Although the bovine embryo elongation has been studied histologically, the essential cellular and molecular factors governing lineage differentiation remain unexplored. This study profiled the transcriptome of single cells in the bovine peri-implantation development throughout day 12, 14, 16, and 18, and identified peri-implantation stage-related features of cell lineages. The candidate regulatory genes, factors, pathways and embryonic and extraembryonic cell interactions were also prioritized to ensure proper embryo elongation in cattle.

Bovine blastocyst-like structures derived from stem cell cultures.

Understanding the mechanisms of blastocyst formation and implantation is critical for improving farm animal reproduction but is hampered by a limited supply of embryos. Here, we developed an efficient method to generate bovine blastocyst-like structures (termed blastoids) via assembling bovine trophoblast stem cells and expanded potential stem cells. Bovine blastoids resemble blastocysts in morphology, cell composition, single-cell transcriptomes, in vitro growth, and the ability to elicit maternal recognition of pregnancy following transfer to recipient cows. Bovine blastoids represent an accessible in vitro model for studying embryogenesis and improving reproductive efficiency in livestock species.

Administering an appeasing substance to beef calves at weaning to optimize productive and health responses during a 42-d preconditioning program.

This experiment evaluated the impacts of administering a bovine appeasing substance (BAS) to beef calves at weaning on their performance, physiological responses, and behavior during a 42-d preconditioning program. Eighty calves (40 heifers and 40 steers; 90% British × 10% Nellore) were weaned at 233 ± 2 d of age (day 0); ranked by sex, weaning age, and body weight (BW); and assigned to receive BAS (IRSEA Group, Quartier Salignan, France; n = 40) or placebo (diethylene glycol monoethyl ether; CON; n = 40). Treatments (5 mL) were topically applied to the nuchal skin area of each animal following dam separation. Within treatment, calves were allocated to one of eight drylot pens (four pens per treatment; pen being the experimental unit) and received a free-choice total mixed ration (TMR) from day 0 to 42, intake of which was assessed daily. Live behavior observations were conducted on days 1, 2, 4, 8, 16, and 32. Temperament was assessed and blood samples were collected via jugular venipuncture on days -21, 0, 3, 7, 14, 28, and 42. Hair samples were collected from the tail switch on days 0, 14, 28, and 42. Calves were vaccinated against bovine respiratory disease viruses on days -21 and 0. Average daily gain from day 0 to 42 did not differ between treatments (P = 0.57) but was greater (P = 0.05) in BAS vs. CON calves from day 0 to 28. Intake of TMR was greater (P = 0.05) during the first week for BAS vs. CON calves (treatment × week; P = 0.08). The mean proportion of calves feeding simultaneously and performance of social and play behaviors were greater (P ≤ 0.05) for BAS vs. CON calves. Escape attempts were greater (P < 0.01) for BAS vs. CON calves on day 1 (treatment × day; P = 0.03). Exit velocity was greater (P = 0.04) for CON vs. BAS calves on day 14 and tended (P = 0.10) to be greater for CON vs. BAS calves on day 7 (treatment × day; P = 0.03). Mean plasma concentrations of haptoglobin were greater (P = 0.02) in CON vs. BAS calves. Hair cortisol concentrations were greater (P = 0.05) in CON vs. BAS calves on day 14 (treatment × day; P = 0.03). Mean serum concentrations of antibodies against bovine viral diarrhea virus were greater (P = 0.02) in BAS vs. CON calves. Collectively, BAS administration to beef calves at weaning alleviated stress-induced physiological reactions, improved temperament evaluated via chute exit velocity, enhanced humoral immunity acquired from vaccination, and appeared to have accelerated adaptation to novel management scheme and environment.

Using low-moisture molasses-based blocks to supplement Ca salts of soybean oil to forage-fed beef cows.

This experiment compared plasma fatty acid (FA) profile of forage-fed beef cows receiving a molasses-based supplement enriched with Ca salts of soybean oil [CSSO; 24.7% of dry matter (DM)] via a self-fed low-moisture block (LMB) or hand-fed granular concentrate daily (CONC). Thirty-six nonlactating, nonpregnant, multiparous beef cows were blocked by age (three blocks), ranked within blocks by body weight (BW) and body condition score (BCS), and allocated to 1 of three drylot pens (27 × 10 m) per block. Nine pens with four cows each were enrolled in a replicated 3 × 2 Latin square design with two periods of 42 d, and a 21-d washout interval. On day 0, pens within each block were randomly assigned to receive one of the three treatments, in a manner that pens did not receive the same treatment in both periods (total n = 6 pens per treatment). Cows received hay (Cynodon dactylon), water, and a mineral-vitamin mix for ad libitum consumption during the study. Hay intake was recorded daily from days 0 to 42, and LMB intake was recorded from days 14 to 42 to allow cows to adapt to supplement with minimal interference from days 0 to 13. The CONC was offered at 0.420 kg/cow daily (DM basis) from days 0 to 13 and then adjusted (days 14 to 42) to match LMB intake. Cow BW and BCS were recorded, and blood samples were collected on days 0, 14, 28, and 42. Average LMB intake during the initial 13 d was 0.846 ± 0.107 kg/cow daily (DM basis). Supplement DM intake did not differ (P = 0.39) between LMB and CONC cows from days 14 to 42 as designed (0.570 vs. 0.583 kg/d, respectively; SEM = 0.011), despite a greater variation in daily intake of LMB vs. CONC (treatment × day interaction; P < 0.01). No treatments effects were noted (P ≥ 0.40) for hay intake, BCS, and BW. Treatment × day interactions were detected (P ≤ 0.01) for plasma concentrations of ω-6 polyunsaturated FA and total FA. On day 0, plasma FA profile did not differ (P ≥ 0.20) between treatments. From days 14 to 42, plasma concentrations of linoleic acid, ω-6 polyunsaturated FA, and total FA were greater (P < 0.01) in CONC and LMB vs. NOSUPP cows. Plasma concentrations of these FA were also greater (P ≤ 0.03) in LMB vs. CONC cows on day 14, but did not differ (P ≥ 0.35) on days 28 and 42. These results indicate that CSSO inclusion into LMB resulted in similar incorporation of ω-6 polyunsaturated and total FA in the circulation compared with CONC offered at the same daily rate. Hence, the use of self-fed LMB appears to be a valid strategy to provide CSSO to forage-fed beef cattle with reduced labor needs.

Supplementing an immunomodulatory feed ingredient to improve thermoregulation and performance of finishing beef cattle under heat stress conditions.

This experiment compared physiological and productive responses in finishing beef cattle managed under heat stress conditions, and supplemented (SUPP) or not (CON) with an immunomodulatory feed ingredient (Omnigen-AF; Phibro Animal Health, Teaneck, NJ). Crossbred yearling cattle (¾ Bos taurus × » Bos indicus; 64 heifers and 64 steers) were ranked by initial body weight (BW) (440 ± 3 kg) and sex, and allocated to 1 of 16 unshaded drylot pens (8 heifers or steers/pen). Pens within sex were randomly assigned to receive SUPP or CON (n = 8/treatment). Cattle received a total-mixed ration (91% concentrate inclusion and 1.21 Mcal/kg of net energy for gain; dry matter [DM basis]) during the experiment (day 0 to 106). The immunomodulatory feed was offered as a top-dress to SUPP pens (56 g/d per animal; as-fed basis) beginning on day 7. Cattle BW were recorded on day 0, 14, 28, 42, 56, 70, 84, 98, and 106. Feed intake was evaluated from each pen by recording feed offer daily and refusals biweekly. Intravaginal temperature of heifers was recorded hourly from day 1 to 6, 29 to 41, and 85 to 97. Environmental temperature humidity index (THI) was also recorded hourly throughout the experiment, and averaged 79.8 ± 0.6. Concurrently with BW assessment, hair samples from the tail-switch were collected (3 animals/pen) for analysis of hair cortisol concentrations. Blood samples were collected on day 0, 28, 56, 84, and 106 from all animals for plasma extraction. Whole blood was collected on day 0, 56, and 106 (3 animals/pen) for analysis of heat shock protein (HSP) 70 and HSP72 mRNA expression. Cattle were slaughtered on day 107 at a commercial packing facility. Results obtained prior to day 7 served as independent covariate for each respective analysis. Heifers receiving SUPP had less (P ≤ 0.05) vaginal temperature from 1500 to 1900 h across sampling days (treatment × hour, P < 0.01; 39.05 vs. 39.19 °C, respectively; SEM = 0.04), when THI ranged from 85.3 to 90.1. Expression of HSP70 and HSP72 was less (P ≥ 0.03) for SUPP cattle on day 106 (22.6- vs. 51.5-fold effect for HSP70, SEM = 9.7, and 11.0- vs. 32.8-fold effect for HSP72; treatment × day, P ≤ 0.04). No treatment effects were detected (P ≥ 0.22) for performance, carcass traits, plasma concentrations of cortisol and haptoglobin, or hair cortisol concentrations. Results from this study suggest that SUPP ameliorated hyperthermia in finishing cattle exposed to heat stress conditions, but such benefit was not sufficient to improve productive responses.