RESUMO
We tested the hypothesis that the biosensor capability of the endometrium is mediated in part, by the effect of different cargo contained in the extracellular vesicles secreted by the conceptus during the peri-implantation period of pregnancy. We transferred Bos taurus taurus embryos of different origin, in vivo (high developmental potential (IV)), in vitro (intermediate developmental potential (IVF)), or cloned (low developmental potential (NT)), into Bos taurus indicus recipients. Extracellular vesicles (EVs) recovered from Day 16 conceptus-conditioned medium were characterized and their microRNA (miRNA) cargo sequenced alongside RNA sequencing of their respective endometria. There were substantial differences in the endometrial response to in vivo versus in vitro and in vivo versus cloned conceptuses (1153 and 334DEGs respectively) with limited differences between in vitro Vs cloned conceptuses (36 DEGs). The miRNA cargo contained in conceptus-derived EVs was similar between all three groups (426 miRNA in common). Only 8 miRNAs were different between in vivo and cloned conceptuses, while only 6 miRNAs were different between in vivo and in vitro-derived conceptuses. Treatment of endometrial epithelial cells with mimic or inhibitors for miR-128 and miR-1298 changed the proteomic content of target cells (96 and 85, respectively) of which mRNAs are altered in the endometrium in vivo (PLXDC2, COPG1, HSPA12A, MCM5, TBL1XR1, and TTF). In conclusion, we have determined that the biosensor capability of the endometrium is mediated in part, by its response to different EVs miRNA cargo produced by the conceptus during the peri-implantation period of pregnancy.
Assuntos
Endométrio , Vesículas Extracelulares , MicroRNAs , Feminino , Endométrio/metabolismo , Endométrio/citologia , Animais , Vesículas Extracelulares/metabolismo , MicroRNAs/metabolismo , MicroRNAs/genética , Bovinos , Gravidez , Técnicas Biossensoriais/métodos , Implantação do Embrião/fisiologia , Embrião de Mamíferos/metabolismoRESUMO
In vivo- and in vitro-produced bovine embryos have different metabolic profiles and differences in gene transcription patterns. These embryos also have a distinct ability to establish and sustain early pregnancies. Small extracellular vesicles (sEVs) are secreted by embryos and carry bioactive molecules, such as miRNAs. We hypothesize that in vivo or in vitro-produced bovine hatched blastocysts on Day 9 and the sEVs secreted by them have different miRNA profiles. To address this hypothesis, embryos of both groups were placed in in vitro culture on Day 7. After 48 h, hatched embryos and hatched embryo-conditioned media (eCM) of both groups were collected. A total of 210 miRNAs were detected in embryos of both groups, of these 6 miRNAs were downregulated, while 7 miRNAs were upregulated in vitro group when compared to in vivo group. sEVs were isolated from eCM to determine miRNA profile. A total of 106 miRNAs were detected in both groups, including 14 miRNAs upregulated in sEVs from in vivo-eCM, and 2 miRNAs upregulated in sEVs from in vitro-eCM. These miRNAs express in embryos and sEVs secreted by them regulate early embryonic developmental and endometrial pathways, which can modify embryo-maternal communication during early pregnancy and consequently affect pregnancy establishment.
Assuntos
Vesículas Extracelulares , MicroRNAs , Animais , Blastocisto/metabolismo , Bovinos , Técnicas de Cultura Embrionária , Embrião de Mamíferos/metabolismo , Desenvolvimento Embrionário/genética , Vesículas Extracelulares/metabolismo , Feminino , MicroRNAs/genética , MicroRNAs/metabolismo , GravidezRESUMO
Wet distiller's grains with solubles (WDGS) are a common by-product feedstuff generated by the grain-ethanol industry, and it is used extensively by the cattle feeding industry. Distillers grains are typically high in protein; however, the protein in WDGS has a low ruminal degradability, and thus may result in a deficiency of RDP in the diet even when dietary CP concentrations are high. Assessment of the RDP needs in diets containing WDGS is needed to aid the cattle feeding industry in managing feed costs and potential environmental issues. To that end, we conducted 2 feeding studies to evaluate the supplemental RDP requirements of beef cattle fed steam-flaked corn-based finishing diets. In Exp. 1, 525 yearling steers (initial body weight = 373 ± 13 kg) received treatments in a 2 × 3 + 1 factorial. Dietary factors included WDGS (15 or 30% of DM) and nonprotein N (NPN; 0, 1.5, or 3.0% of DM) from urea (0, 0.52, and 1.06%). The control diet without WDGS contained 3.0% NPN (1.06% urea) and cottonseed meal. Diets were formulated to have equal crude fat concentrations. Overall gain efficiency among steers fed 15% WDGS was greatest for 1.5% NPN and least for 0% NPN (P = 0.07, quadratic), whereas gain efficiency decreased linearly (P < 0.09) as NPN increased in the 30% WDGS diets. Dressing percent was greater (P < 0.01) for the Control diet than for 15 or 30% WDGS. In Exp. 2, 296 steer calves (initial BW = 344 ± 12 kg) were fed 1 of 4 experimental diets that included a Control diet without WDGS (contained 3% NPN from urea, and cottonseed meal) and 15% WDGS diets with either 1.50, 2.25, or 3.00% NPN (0.52, 0.78, and 1.04% urea, respectively, on a DM basis). Overall gain efficiency on either a live or carcass-adjusted basis was not different among treatments (P > 0.15). Dietary NPN concentration did not influence growth performance (P > 0.21). Increasing dietary WDGS concentration resulted in decreasing (P < 0.05) diet digestibility (determined with an internal marker) and increasing (P < 0.05) N volatilization losses (determined by diet and manure N:P ratio); however, the effects of NPN level on digestibility and N losses were somewhat inconsistent across experiments. Results suggest that optimum performance for cattle fed 15% WDGS occurred when the diet contained between 1.5 and 2.25% NPN. However, no supplemental NPN was needed to support optimum performance in diets containing 30% WDGS.
Assuntos
Ração Animal/análise , Bovinos/fisiologia , Suplementos Nutricionais , Nitrogênio/metabolismo , Animais , Peso Corporal , Bovinos/crescimento & desenvolvimento , Dieta/veterinária , Digestão , Grão Comestível , Cavalos , Masculino , Nutrientes , Vapor , Ureia/metabolismo , Zea maysRESUMO
The use of distiller's grains (DG) in beef cattle finishing diets is a common practice. However, the effects of supplemental fat on performance and nutrient losses of cattle fed diets containing DG are not known. Therefore, we fed 398 crossbred yearling heifers (initial BW = 373.5 kg) for 106 d to determine the effects of dietary fat concentration and sorghum-based wet distiller's grains with solubles (SWDGS) on performance, carcass characteristics, and nutrient losses of finishing cattle. Treatments included two 92% concentrate, steam-flaked corn (SFC)-based diets with 0% or 3% added fat from yellow grease and 3 SFC-based diets with 15% SWDGS (DM basis) that contained either 0%, 1.5%, or 3% added fat (8 pens per treatment) in a randomized block design. Overall DMI and ADG were 5% to 6% greater (P < 0.01) for heifers fed 15% SWDGS than for those fed 0% SWDGS. Among heifers fed 15% SWDGS, DMI was greatest (P = 0.04; quadratic effect) and ADG tended (P = 0.12; quadratic effect) to be greatest for heifers fed 1.5% fat. The ADG:DMI did not differ between 0% SWDGS with 0% or 3% fat, and was not altered by replacing a portion of SFC with SWDGS (P > 0.36). However, ADG:DMI tended to increase as more fat was added to diets with 15% SWDGS (P = 0.06). Average hot carcass weight (HCW) was 5 kg greater (P = 0.05) when SWDGS was fed, but HCW tended to be greatest for heifers fed 15% SWDGS with 1.5% fat (P = 0.09, quadratic effect). Heifers fed 0% SWDGS with 0% fat tended to have a lower marbling score, less rib fat, lower average yield grade (P < 0.08), and more (P < 0.01) yield grade 1 carcasses than heifers fed 0% SWDGS with 3% fat. Averaged across fat levels, heifers fed 15% SWDGS had more rib fat and a higher yield grade (P < 0.03) than heifers fed 0% SWDGS. Feeding 15% SWDGS did not alter carcass quality grade distribution compared to feeding 0% SWDGS, but 15% SWDGS produced fewer yield grade 3 carcasses (P = 0.03) than 0% SWDGS. The calculated NEg of SWDGS (1.36 Mcal/kg) was 91% of the tabular value for dry rolled corn (1.50 Mcal/kg) and 84% of the tabular value for SFC (1.62 Mcal/kg). Nitrogen intake, and N excretion were greater (P < 0.05) in heifers fed 15% SWDGS than in heifers fed the 0% SWDGS diets, but N loss as a % of N intake was less (P < 0.05). Our results suggest adding 1.5% fat to diets containing 15% SWDGS may improve beef cattle performance; however, feeding logistics need to be considered when pricing wet DG.