Influence of estradiol on bovine trophectoderm and uterine gene transcripts around maternal recognition of pregnancy

Embryo survival and pregnancy success is increased among animals that exhibit estrus prior to fixed time-artificial insemination, but there are no differences in conceptus survival to d16. The objective of this study was to determine effects of preovulatory estradiol on uterine transcriptomes, select trophectoderm (TE) transcripts, and uterine luminal fluid proteins. Beef cows/heifers were synchronized, artificially inseminated (d0), and grouped into either high (highE2) or low (lowE2) preovulatory estradiol. Uteri were flushed (d16); conceptuses and endometrial biopsies (n = 29) were collected. RNA sequencing was performed on endometrium. Real-time polymerase chain reaction (RT-PCR) was performed on TE (n = 21) RNA to measure relative abundance of IFNT, PTGS2, TM4SF1, C3, FGFR2, and GAPDH. Uterine fluid was analyzed using 2D Liquid Chromatography with tandem mass spectrometry-based Isobaric tags for relative and absolute quantitation (iTRAQ) method. RT-PCR data were analyzed using the MIXED procedure in SAS. There were no differences in messenger RNA (mRNA) abundances in TE, but there were 432 differentially expressed genes (253 downregulated, 179 upregulated) in highE2/conceptus versus lowE2/conceptus groups. There were also 48 differentially expressed proteins (19 upregulated, 29 downregulated); 6 of these were differentially expressed (FDR \u3c 0.10) at the mRNA level. Similar pathways for mRNA and proteins included: calcium signaling, protein kinase A signaling, and corticotropin-releasing hormone signaling. These differences in uterine function may be preparing the conceptus for improved likelihood of survival after d16 among highE2 animals

Effects of preovulatory estradiol on uterine environment and conceptus survival from fertilization to maternal recognition of pregnancy

Preovulatory estradiol is known to impact embryo quality and survival. The objective of this study was to determine the effects of preovulatory estradiol on the uterine environment and conceptus survival through maternal recognition of pregnancy. Beef cows/heifers were AIed following induced ovulation. Cows were grouped into high and low preovulatory estradiol. Conceptuses were collected on day 16 nonsurgically (Rep 1; n = 20), or following slaughter (Rep 2; n = 29). Blood was collected to determine plasma glucose concentrations, and uterine luminal fluid (ULF) was analyzed for protein, glucose, and interferon tau (IFNT) concentrations. Total cellular RNA was extracted from caruncular (CAR) and intercaruncular (INCAR) endometrial tissue. There was no effect of preovulatory estradiol on conceptus recovery rate (P = 0.38) or on apoptosis rate in the trophectoderm (P = 0.64). Cows in which a conceptus was recovered had greater concentrations of protein in the ULF (P = 0.04). Animals with elevated preovulatory estradiol had greater endometrial abundance of SLC2A1 (P = 0.05) and SLC5A1 (P = 0.04) in both INCAR and CAR tissue. Presence of a conceptus also tended to increase (P = 0.10) abundance of SLC5A1 in INCAR. In CAR tissue, cows with a conceptus had decreased SLC2A4 abundance (P = 0.05). In summary, conceptus recovery rates, apoptosis in the trophectoderm, IFNT, glucose, and protein concentration in ULF did not differ between cows that did or did not have an increase in preovulatory estradiol concentrations. Thus, there is no indication of increased conceptus survival to day 16 of pregnancy based on estradiol concentrations

Effects of preovulatory estradiol on uterine environment and conceptus survival from fertilization to maternal recognition of pregnancy

Preovulatory estradiol is known to impact embryo quality and survival. The objective of this study was to determine the effects of preovulatory estradiol on the uterine environment and conceptus survival through maternal recognition of pregnancy. Beef cows/heifers were AIed following induced ovulation. Cows were grouped into high and low preovulatory estradiol. Conceptuses were collected on day 16 nonsurgically (Rep 1; n = 20), or following slaughter (Rep 2; n = 29). Blood was collected to determine plasma glucose concentrations, and uterine luminal fluid (ULF) was analyzed for protein, glucose, and interferon tau (IFNT) concentrations. Total cellular RNA was extracted from caruncular (CAR) and intercaruncular (INCAR) endometrial tissue. There was no effect of preovulatory estradiol on conceptus recovery rate (P = 0.38) or on apoptosis rate in the trophectoderm (P = 0.64). Cows in which a conceptus was recovered had greater concentrations of protein in the ULF (P = 0.04). Animals with elevated preovulatory estradiol had greater endometrial abundance of SLC2A1 (P = 0.05) and SLC5A1 (P = 0.04) in both INCAR and CAR tissue. Presence of a conceptus also tended to increase (P = 0.10) abundance of SLC5A1 in INCAR. In CAR tissue, cows with a conceptus had decreased SLC2A4 abundance (P = 0.05). In summary, conceptus recovery rates, apoptosis in the trophectoderm, IFNT, glucose, and protein concentration in ULF did not differ between cows that did or did not have an increase in preovulatory estradiol concentrations. Thus, there is no indication of increased conceptus survival to day 16 of pregnancy based on estradiol concentrations

Differences in Relative Abundance of GnRH-I and GnRH-II in Granulosa Cells of Bovine Antral Follicles at Specific Stages of Follicular Development

Increased estradiol is the primary signal to initiate standing estrus, and cows that exhibit estrus prior to fixed-time artificial insemination have greater pregnancy rates than cows that do not. Therefore, understanding what factors may be present at the ovary that may regulate estradiol production is critical. A previous study reported that bovine follicles with greater follicular fluid concentrations of estradiol had decreased expression of GnRH-I and GnRH-II in granulosa cells (GC). The objective of this study was to characterize relative abundance of GnRH-I and -II mRNA within GC of follicles at specific stages of development. Beef cows were synchronized, and ovaries were collected at specific stages of follicular development [pre-selection (PRE), post-selection (POST), and post-selection 24 h after luteal regression (POST-PG)]. All surface follicles were classified as small (\u3c 5mm), medium (5 - 8mm), or large (\u3e 8mm) and aspirated to collect GC. Large follicles from each animal were kept separate and all other follicles were pooled by size within animal (n = 27, 27, and 18 for small, medium, and large). Total cellular RNA was extracted, and RT-PCR was performed for GnRH-I, GnRH-II, and GAPDH. Data were analyzed using the MIXED procedure of SAS. Across all follicles, GnRH-I and GnRH-II expression were not influenced by stage (P = 0.27) but were influenced by size (P \u3c 0.01). Smalls (4.55 ± 0.39 and 3.91 ± 0.44, respectively) had greater expression (P ≤ 0.01) of GnRH compared to mediums (0.83 ± 0.39 and 1.41 ± 0.44, respectively) and larges (0.52 ± 0.47 and 2.12 ± 0.54, respectively). There was also a stage by size interaction (P \u3c 0.01). POST (P \u3c 0.01) and POST-PG (P ≤ 0.08) smalls had or tended to have increased expression compared to PRE smalls, but PRE mediums had increased expression (P \u3c 0.03) compared to POST-PG mediums. When only the largest follicle for each animal was evaluated, stage of development influenced expression of GnRH-I (P = 0.03) but not GnRH-II (P = 0.91). For GnRH-I, PRE tended (P = 0.09; 2.29 ± 0.55) to have increased expression compared to POST (0.92 ± 0.55) and did have greater expression compared to POST-PG (P = 0.01; 0.11 ± 0.55). Thus, GnRH within antral follicles may be a key regulator of the follicle’s ability to produce estradiol

Influence of Bovine Viral Diarrhea Virus Infection on Artificial Insemination Conception and Breeding Season Pregnancy Success in Vaccinated Beef Herds

Bovine Viral Diarrhea Virus (BVDV) causes reproductive economic losses in cattle. The objective of this study was to evaluate the influence of BVDV infection on reproductive success. Vaccinated cows (n = 370) and heifers (n = 528) from nine different herds were synchronized using the 7-day CO-Synch + CIDR protocol and were bred using fixed-time artificial insemination (FTAI). On d 28 following insemination, blood samples were collected and pregnancy status was determined. Non-pregnant animals were resynchronized and FTAI occurred a second time. In six herds, bulls were comingled with females beginning 10-15 d after the second AI. Final pregnancy status was determined 33-80 d following the first pregnancy diagnosis. Blood samples were tested for the presence of BVDV antigen using the IDEXX BVDV PI X2 Kit. Animals that tested positive were considered infected with BVDV at the time of blood collection. Herds were determined to be BVDV infected by the presence of at least one animal having a positive test for antigen (n = 4 infected herds, n = 5 non-infected herds). Statistical analyses were performed using the GLIMMIX procedure of SAS with herd as a random variable. Herds that had evidence of BVDV infection at d 28 following insemination had significantly decreased (P \u3c 0.01) first service AI conception rates compared to herds that had no evidence of infection (34 ± 2.3% vs. 54 ± 2.3%, respectively). Additionally, breeding season pregnancy rates were decreased (P \u3c 0.01) in BVDV infected herds compared to non-infected herds (69 ± 3.4% vs. 80 ± 3.6%, respectively). There was no significant effect of BVDV infection status on embryonic loss (P = 0.42) or percentage of animals which lost a pregnancy and rebred by the end of the breeding season (P = 0.63). In conclusion, BVDV infection in well vaccinated herds had a significant negative impact on both first service AI conception rate and overall breeding season pregnancy success

Greater numbers of antral follicles in the ovary are associated with increased concentrations of glucose in uterine luminal fluid of beef heifers

Increased antral follicles are associated with greater fertility and a uterine environment that is more supportive of early embryonic development in beef heifers. Glucose is a primary energy source for embryos, and glucose concentrations are elevated in uterine luminal fluid (ULF) of pregnant heifers. We hypothesized that ULF glucose concentrations and endometrial transcript abundance for glucose transporters on d16 after insemination would be greater in heifers with increased numbers of antral follicles. Heifers classified with either increased or diminished antral follicle counts were artificially inseminated following the CO-Synch protocol (d0). On d16 after insemination, reproductive tracts of heifers were collected at an abattoir to retrieve conceptuses to determine pregnancy. Uterine luminal fluid was collected, endometrium was biopsied, total RNA was extracted and glucose transporter transcript abundances were determined. Data were analyzed using the MIXED procedure of SAS with antral follicle group, pregnancy status, and the interaction as fixed effects. Glucose concentrations in ULF were greater in heifers with increased antral follicle numbers. Glucose ULF concentrations increased in pregnant heifers. Facilitated glucose transporter member 1 (SLC2A1) transcript abundance was increased in the endometrium of pregnant heifers but was not different due to antral follicle number or the interaction. Differences in uterine concentrations of glucose associated with antral follicle number could be due to another mechanism, since glucose transporters were not different between antral follicle numbers. Therefore, heifers with increased number of antral follicles have increased energy availability in the uterus to support trophoblast proliferation and function

Brangus cows have ovarian reserve parameters more like Brahman than Angus cows

Bos indicus females have more surface antral follicles than Bos taurus females; however, histological studies demonstrated no difference in total number of primordial follicles between these two biological types of cattle. Primordial follicle density in the ovary was less in Nelore ovaries compared to Angus ovaries, but no studies have examined the primordial follicle density in Bos indicus cross-bred females. It, therefore, was hypothesized that primordial follicle density in the ovary would decrease as percentage Bos indicus increased. Ovaries were collected from cross-bred Angus (n=32, no Bos indicus influence), Brangus (n=15), or Brahman (n=9) cows and prepared for histological evaluation. There was no difference in total number of primordial follicles per ovary between breeds (P \u3e 0.10). When numbers of primordial follicles were expressed on a per gram of ovarian tissue basis, there were fewer primordial follicles per gram of ovarian tissue in Brangus and Brahman cows than in Angus cows (P \u3c 0.05). Brangus cows did not differ from Brahman cows in primordial follicle density (P \u3e 0.10). Differences in primordial follicle density could indicate differences in capacity of ovarian stroma to produce factors necessary for oogonial proliferation and primordial follicle formation among breeds. Identifying these factors could improve the aprroach for culturing pre-antral follicles of cattle. Furthermore, these results explain why ultrasonographic antral follicle counts may need to be adjusted to a greater threshold to predict size of the ovarian reserve and determine ovarian reserve related reproductive traits in Bos indicus females

Higher education and sustainable development of marine resources

Marine park is a designated area of the sea where national regulatory authorities place limits on certain human activities. Marine parks usually allow multiple uses. Certain zones are protected for preserving specific habitats for marine biodiversity to achieve ecological sustainability while in others activities such as recreational fishing, ecotourism, snorkeling, and diving are allowed. Large marine parks have open zones for fishing and no-take zones where extractive activities such as fishing, sand mining, and drilling are prohibited. Generally, marine parks are considered suitable places for gaining scientific understanding and promoting environmental awareness while safeguarding the benefits of indigenous coastal communities. Sea ranching is a type of marine farming in which juveniles are released into the ocean where they grow without containment structures. The juveniles are generally produced in the hatcheries. In some cases, wild-caught juveniles from one area are also released into another area for stock enhancement or creating a fishery resource. They grow unprotected and without any assistance such as feed from outside. The marine environment provides the juveniles all the resources to grow to be subsequently harvested. Sea ranching is unlike mariculture where stocked species are held in captivity, in cages, sea pens, or other types of enclosures, and provided feed and other requirements from external sources. Nature-based solutions are human actions aimed at sustainably managing the natural or modified ecosystems. Such solutions include measures to protect and restore the ecosystems and are inspired by nature. In the context of oceans, the nature-based solutions are systematic and informed interventions that are resource-efficient and help in building resilience. This concept is based on the understanding that healthy ocean provides more ecosystem services that benefit the humanity