To evaluate the effect of conditioned media (CM) and Extracellular Vesicles (EVs) derived from bovine oviduct epithelial cell (BOEC) lines on the developmental capacity of bovine zygotes and the quality of embryos produced in vitro, presumptive zygotes were cultured under specific conditions. In experiment 1, zygotes were cultured either on monolayers from BOEC extended culture (E), together with fresh BOEC suspension cells, or with BOEC-CM from fresh or E-monolayers. In experiment 2, EVs were isolated from BOEC-CM and characterized (150–200 nm) by Nanosight^® and electron microscopy. Zygotes were cultured in the presence of 3x10⁵EVs/mL, 1.5x10⁵EVs/mL or 7.5x10⁴EVs/mL of fresh or frozen BOEC-EVs. In experiment 3, zygotes were cultured in absence of FCS but with EVs from BOEC-E that had been cultured in different culture media. In experiment 4, zygotes were cultured in SOF+5% normal-FCS, or EV-depleted-FCS. In all cases, cleavage rate (Day 2) and blastocyst development (Day 7–9) was assessed. Blastocysts on Days 7/8 were used for quality evaluation through differential cell count, cryotolerance and gene expression patterns. No differences were found among all FCS-containing groups in cleavage rate or blastocyst yield. However, embryos derived from BOEC-CM had more trophectoderm cells, while embryos derived from BOEC-EVs, both fresh and frozen, has more trophectoderm and total cells. More embryos survived vitrification in the BOEC-CM and BOEC-EV groups. In contrast, more embryos survived in the EV-depleted-FCS than in normal-FCS group. Gene expression patterns were modified for PAG1 for embryos cultured with EVs in the presence of FCS and for IFN-T, PLAC8, PAG1, CX43, and GAPDH in the absence of FCS. In conclusion, EVs from FCS have a deleterious effect on embryo quality. BOEC-CM and EVs during in vitro culture had a positive effect on the quality of in vitro produced bovine embryos, suggesting that EVs have functional communication between the oviduct and the embryo in the early stages of development.

Bovine beta-defensin 126 (BBD126) exhibits preferential expression for the cauda epididymis of males, where it is absorbed onto the tail and postacro

While the removal of seminal plasma is a routine practice prior to equine sperm cryopreservation, this fluid contains the main source of antioxidant enzymes able to scavenge these reactive oxygen species. Therefore, stallion seminal plasma components may have an impact on ejaculate freezability. Against this background, this study was designed to investigate whether the activities of the main stallion seminal plasma antioxidant enzymes are related to sperm cryotolerance. With this purpose, 16 ejaculates were collected from 14 healthy stallions, and each ejaculate was split into two aliquots. The first one was used to evaluate the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), and glutathione reductase (GSR) in seminal plasma. The second aliquot was extended and then processed for cryopreservation. Sperm motility and viability were evaluated before and after cryopreservation, and ejaculates were classified as of good (GFE) or poor freezability (PFE) based on total motile and viable spermatozoa at post-thaw. We observed that, while the specific activities of CAT, GPX, and GSR were similar between GFE and PFE, that of SOD was significantly (p < 0.05) higher in GFE than in PFE. We can thus conclude that, in stallions, the specific activity of SOD in the seminal plasma of a given ejaculate might be related to its freezability.

The aim was to examine the effect of embryo–uterine synchrony on conceptus elongation and pregnancy rate in cattle. In Study 1, crossbred beef heifers each received 10 Day-7 in vitro-produced blastocysts on either Day 5, 7 or 9 after oestrus. A proportion of Day 5 recipients were supplemented with progesterone, via a progesterone-releasing intravaginal device from Days 3–5 plus either 750 IU equine chorionic gonadotrophin or 3000 IU human chorionic gonadotrophin on Day 3. At embryo age Day 14, all heifers were slaughtered and the uterus was flushed. Fewer recipients yielded conceptuses (P < 0.05) and fewer conceptuses were recovered (P < 0.05) following transfer on Day 5 compared with Day 7 or 9. Supplementation with progesterone resulted in short cycles in approximately 50% of recipients. Mean conceptus length was greater (P < 0.05) following transfer to an advanced uterus. In Study 2, overall pregnancy rate following the fresh transfer of a single in vitro-produced blastocyst was 43.5% (2065/4749). Transfer of a Day 7 embryo to a synchronous Day-7 uterus resulted in a pregnancy rate of 47.3%. Transfer to a Day-5 (40.8%) or a Day-8 (41.3%) uterus moderately impacted pregnancy rate (P < 0.01) while transfer to a uterus 2 days in advance (Day-9, 24.4%) or 3 days behind (Day-4, 27.0%) reduced (P < 0.001) pregnancy rate compared with synchronous transfers. In conclusion, transfer of an embryo into an advanced uterus results in an acceleration of conceptus development, but does not result in greater pregnancy rates.