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Small equine embryos, equal or smaller than three hundred micrometers in diameter (≤ 300 μmØ) are collected before the seventh post ovulation (PO) day. Big embryos, larger than three hundred micrometers (>300 μmØ) are recovered during or after the seventh PO day. No experiments test the type of holding and washing solutions and to what extent they influence the embryo post vitrification survival. Further knowledge is needed about the mixture of cryoprotectants; either penetrating or non penetrating and the effect they exert over the embryos post vitrification survival and pregnancy rate (PVSPR). The type of embryo carrier, the size of the embryo and the volume of the vitrification solution vary between small and large embryos. A high temperature transfer index (TTI) optimizes the embryo PVSPR. In equids, the type of holding and transfer solutions used during the post vitrification step is an area scarcely explored. This study endeavors to provide information to help in the adaptation of vitrification protocols depending on the embryo size. An additional objective is to ease the access to data about the solutions employed before and during the vitrification; as well as the solutions used during the embryo warming and transfer steps. [ABSTRACT FROM AUTHOR]

Background: Proper timing for embryo collection and transfer in horses -which is critical for the success of this biotechnology- is still debated. Additionally, there is little information on this technology under tropical conditions. Objective: To determine the best day for collection and transfer of embryos in Mangalarga Marchador mares under Brazilian northeast’s conditions. Methods: Donors (n= 30) and recipients (n= 76) in diestrus phase were selected based on both clinical and gynecology examinations. Estrus was induced on both donor and recipient mares by intramuscular injection of 5 mg Dinoprost, aiming to obtain an ovulation interval of -1 to +3 between recipient and donor. Ovulation was induced with buserelin acetate when the largest follicle reached at least 35 mm in diameter. At this time, mares were subjected to artificial insemination at 48-hour intervals until ovulation. The embryos were collected on days 7, 8, and 9 after ovulation. Results: The embryo collection on day 8 was more efficient (p0.05) than day 9, which presented the same efficiency (p>0.05) as day 7. From a total of 76 embryos transferred to the recipients, that were between days 4 and 9 after ovulation, there was no influence (p>0.05) of the day of transfer on pregnancy rate. Conclusions: The embryo collection must be performed on day 8 after ovulation, and transfer can be performed on any day of that interval (4-9) without affecting the pregnancy rate.

A study was conducted to investigate whether ovulation in gilts could be synchronized for embryo collection by the administration of estradiol benzoate (EB) or estradiol dipropionate (EDP) to induce pseudopregnancy, followed by the treatment with prostaglandin F2α (PGF2α) on 10 days after. Ten gilts each received a total of 20 mg of EB or EDP on Day 10 or EB on Day 10 and 14 to induce pseudopregnancy (Day 0 = onset of estrus). Donors received PGF2α 10 or 15 days (as a control) after the first administration of estrogens and subsequently eCG and hCG, and were then inseminated artificially. The embryos were collected 7 days after the administration of hCG, and assessed for embryo yield and their developmental stages. All protocols resulted in good embryo yield (9.8-13.2 embryos in average), and the embryos showed average ability to develop to the expanded blastocyst stage (3.29-4.03 as developmental scores) without any significant differences among the protocols. These results suggest that the administration of PGF2α 10 days after the treatment of gilts with EB or EDP would allow synchronization of ovulation and embryo collection, as well as shortening the period from estrus detection to embryo collection, thus improving embryo collection efficiency. [ABSTRACT FROM AUTHOR]

Despite a long history of bovine superovulation research, significant commercial applications did not start until the early 1970s. For some 20 years thereafter, superovulation represented the primary tool for the production of cattle embryos. In the early 1990s, commercial in vitro production (IVP) was initiated in cattle. Although ovum pick-up and IVP are now commercially practiced on a wide scale, superovulation and embryo recovery by flushing remain a widespread and very effective approach to the production of cattle embryos. This review covers both the history and the effects of multiple factors on superovulation in Bos taurus cattle. There are three general protocols for suitable pre-FSH programming of donors so that gonadotrophin-responsive follicles are available. Superovulation protocols vary widely based on the FSH source, the diluent used, the number and timing of FSH injections and the timing and utilisation of various prostaglandins, controlled internal progesterone releasing devices, gonadotrophin-releasing hormone, and other means of controlling follicular development and ovulation. The number of oocytes that can be stimulated to grow and ovulate within any given donor can be estimated by either ultrasound-guided sonography or by measuring concentrations of anti-Müllerian hormone in the blood. Animal-related factors that can influence the efficacy of superovulation include cattle breed, age, parity, genetics, lactational status and reproductive history. In addition, nutrition, stress, season, climate, weather and several semen factors are discussed. [ABSTRACT FROM AUTHOR]

An efficient system to collect large numbers of vital zygotes is a pre-requisite for application of zygote genome-editing technology, including development of efficient models for xenotransplantation using pigs. Owing to the sub-optimal in vitro production of zygotes in pigs, efficient collection of in vivo developed zygotes is required. Timing of ovulation is a key factor to sustain efficiency since the interval between pronuclear formation and the first division is very short in pigs. The weaning-to-estrus interval can, due to its inverse relation with length of estrus and time of ovulation, interfere with ovulation and make it asynchronous, which reduces the probability of obtaining zygotes. This retrospective study compared the effects of three weaning-to-estrus intervals of 3, 4 or 5 days on zygote collection efficiency in a total of 17 trials over a 3-year period including 223 sows. Donor sows in groups of 10–15 animals were super-ovulated with eCG 24 h after weaning and those in estrus at 48–72 h post-eCG were immediately treated with hCG, followed by insemination 6 and 24 h thereafter. Collected structures during laparotomy on Day 2 (Day 0: onset of estrus) were morphologically evaluated and only those with a single cell and two visible polar bodies were considered as zygotes. Zygotes were injected with CRISPR-Cas9 editor mixture and cultured for 6 days to evaluate their developmental ability against non-injected control zygotes. Of all recovered structures (N = 5,468), 67.4%, 30.8% and 1.8% were zygotes, 2-cell embryos and oocytes-degenerated embryos, respectively. The different weaning-to-estrus intervals did not affect either the percentages of collected zygotes (range: 64.1%–70.0%) or the percentages of sows with zygotes at collection time (range: 69.0%–73.3%). The weaning-to-estrus intervals did not affect the in vitro developmental ability of zygotes. After 24 h of culture, 78.1 ± 2.0% and 95.1 ± 0.6 (P < 0.05) of injected (N = 2,345) and non-injected (N = 335) zygotes, respectively, developed to 2-to-4-cell embryo stage. The total efficiency of the system was 64.1 ± 2.2% and 85.8 ± 1.5% (P < 0.05) for injected and non-injected zygotes, respectively. In conclusion, the results indicate that neither the efficiency of collecting in vivo derived porcine zygotes from superovulated sows nor the zygote ability to develop to blastocyst after cytoplasmic genome-editing injection were affected by a weaning-to-estrus interval between 3-to-5 days. • Genome-editing technology requires efficient methods to obtain large numbers of vital zygotes. • We describe a protocol for collecting high-quality porcine zygotes from weaned sows. • Weaning-to estrus intervals between 3 and 5 days did not affect the zygote collection efficiency. • These intervals did not affect the developmental ability of CRISPR-Cas9 injected zygotes. [ABSTRACT FROM AUTHOR]

We aimed to define whether embryo collection carried out after pseudopregnancy was of similar outcome and quality as after artificial abortion. To induce pseudopregnancy, 30 gilts or sows were given 20 mg intramuscular estradiol dipropionate (EDP) 10–11 days after the onset of estrus. Ten additional pigs were inseminated artificially at natural estrus as a control group. Prostaglandin F2α (PGF2α) was administered twice with a 24 hr interval beginning 15, 20, or 25 days after EDP‐treatment (n = 10 per group) or between 23 and 39 days after artificial insemination in control pigs. Following this, all pigs were given 1,000 IU equine chorionic gonadotropin and 500 IU human chorionic gonadotropin (hCG) and then inseminated. Embryos were recovered 6 or 7 days after hCG treatment and outcome was recorded. There was no significant difference in the number of normal embryos collected from the pigs with PGF2α initiated at different time points or from the control group. Embryonic developmental stages 7 days after hCG treatment also did not differ among groups. These results indicate that the use of EDP to induce pseudopregnancy, followed by PGF2α administration to synchronize estrus for subsequent embryo harvest, is a suitable alternative to the artificial abortion method. [ABSTRACT FROM AUTHOR]

Embryo transfer (ET) could be a relevant tool for genetic improvement programs in horses similar to those already underway in other species and produce multiple foals from the same mare in one breeding season. However, there have been no reports describing equine embryo transfer performed in Korea. In the present study, we performed an equine embryo collection and transfer procedure for the first time. We examined the embryo collection and pregnancy, size of embryo during the incubation period after collection, and progesterone (P4) and estradiol-17ß (E2) concentrations in mare’s serum at embryo collection and transfer. A total of 16 donors responded to estrus synchronization; estrus was induced in 12 donors and 4 recipients, and artificial insemination was successful in 10 donors and six blastocysts were collected from donors. Of these blastocysts, we monitored the size of blastocysts for 3 day during incubation and transferred 2 blastocysts to a recipient, with 1 successful pregnancy and foal achieved. The dimensions of equine embryo at day 7 to day 9 were 409 μm, 814 μm and 1,200 μm. The serum P4 and E2 concentrations were 7.91±0.37 ng/μL and 45.45±12.65 ng/μL in the donor mare, and 16.06±3.27 ng/μL and 49.13±10.09 ng/μL in the recipient mare.

Single-cell genomics allow the characterization and quantification of molecular heterogeneity from a wide variety of tissues. Here, we describe the manual dissociation and collection of single cells, a method adapted for the characterization of precious small tissues like preimplantation embryos. We also describe the acquisition of mouse embryos by flushing of the oviducts. The cells can then be used in multiple sequencing protocols, for example, Smart-seq2, Smart-seq3, smallseq, and scBSseq., (© 2023. Springer Science+Business Media, LLC.)

Alpacas can only produce one offspring per year. In order to accelerate the genetic gain of a herd, superovulation and embryo transfer can be used to produce multiple embryos from superior females. We hypothesized that the use of dual siring with superovulation would result in the production of multiple embryos sired by different males. After administration of the superovulation protocol, receptive females were bred to two proven males (A and B) 8–12 h apart and ovulation was induced by gonadotropin at the time of the first breeding. Growth of multiple dominant follicles was successfully achieved in 95% of cycles. Females that were receptive after FSH treatment and were bred with both males (order A–B or B–A). Embryo collections were performed 8–9 days post-breeding on 15 cycles and 73% of collections recovered ≥1 embryo. A total of 46 embryos, were recovered for an average of 3.13 ±3.1 (range 0–10) embryos/flush. Parentage analysis was performed for 23 embryos (6 from A to B, 17 from B to A). Twenty-two of the 23 embryos were determined to be sired by male B, being six embryos from breeding A-B and 16 embryos from breeding B-A. A single embryo from breeding B-A was sired by male A. In conclusion, FSH administered at decreasing doses can be used to promote superovulation resulting in collection of multiple embryos per cycle. However, slight differences in male fertility may affect the frequency of embryos sired by each male. [ABSTRACT FROM AUTHOR]

Abstract: Finland blue fox (Alopex lagopus) has great reputation in pelt industry around the world for its large size and top-ranking fur quality; however, both the herd size and the average survival rate of purebred offspring are rather low in production systems in China. Surgical transfer of blue fox embryos was investigated as a means to increase the population fox and also as a possible means to conserve endangered canine species. The animals were chosen on the basis of synchrony in natural oestrus. During the reproductive season of blue fox, 59 embryos were flushed from 6 farmed donors 9–11 days after the first insemination, and 53 embryos were transferred surgically into the uteri of the 6 paired recipients with natural synchronized oestrous. Two of the recipients littered 46–49 days after embryo transfer; one gave birth to 7 pups and the other 1 pup. This report describes the first successful embryo transfer in the farmed blue fox in China. [Copyright &y& Elsevier]

Early return-to-estrus after embryo collection would shorten the interval between consecutive superovulations and improve efficiency of embryo production. Following superovulation and embryo collection, 80 buffaloes were treated with 15.0 mg Luprostiol (PGF2α analogue) for the induction of luteolysis and early return-to-estrus. A total of 67.5% donor animals returned to estrus, on average 11.8±0.84 days after the PGF2α treatment. The number of ovulations (≤5 or >5 CL) had no significant effect on the percentage of donors returning to estrus within 30 days, as 70% of the buffaloes with ≤5 CL and 65% with >5 CL returned to estrus during this time. However, an increase in the number of ovulations significantly delayed the return to estrus as this duration was 9.7±0.93 days in the buffaloes with ≤5 CL compared to 14.1±1.29 days in those having >5 CL. [Copyright &y& Elsevier]

A study was conducted to investigate whether ovulation in gilts could be synchronized for embryo collection by the administration of estradiol benzoate (EB) or estradiol dipropionate (EDP) to induce pseudopregnancy, followed by the treatment with prostaglandin F 2α (PGF 2α ) on 10 days after. Ten gilts each received a total of 20 mg of EB or EDP on Day 10 or EB on Day 10 and 14 to induce pseudopregnancy (Day 0 = onset of estrus). Donors received PGF 2α 10 or 15 days (as a control) after the first administration of estrogens and subsequently eCG and hCG, and were then inseminated artificially. The embryos were collected 7 days after the administration of hCG, and assessed for embryo yield and their developmental stages. All protocols resulted in good embryo yield (9.8-13.2 embryos in average), and the embryos showed average ability to develop to the expanded blastocyst stage (3.29-4.03 as developmental scores) without any significant differences among the protocols. These results suggest that the administration of PGF 2α 10 days after the treatment of gilts with EB or EDP would allow synchronization of ovulation and embryo collection, as well as shortening the period from estrus detection to embryo collection, thus improving embryo collection efficiency., (© 2021 Japanese Society of Animal Science.)

Background: Age and antral follicle count (AFC) are related to fertility in cattle, but this information remains limited in mares., Methods: To verify the influence of age and AFC on the reproductive characteristics of mares, 15 Quarter Horse donors, with 5-15 antral follicles, ranging from three to 17 years old, healthy and in good nutritional status, were divided into groups with low AFC (≤9 follicles) or high AFC (≥10 follicles) and mares considered young (≤9 years) or old (≥10 years). Mares were submitted to ultrasonography to determine the dominant follicle diameter, follicular growth rate, degree of uterine oedema and embryonic recovery for a minimum of three oestrous cycles., Results: AFC was higher (P=0.001) in young mares compared with old mares. An interaction (P=0.001) between AFC and age was observed with regard to follicular growth rate, being that mares with low AFC and are old showed the lowest follicular growth compared with those of low AFC and young, high AFC and old, and high AFC and young. Younger mares and those with high AFC exhibited higher degree of uterine oedema (P<0.05) on the third day of oestrus compared with older mares and with low AFC (3 and 2, respectively). However, in both groups, the degree of oedema reduced by the time of ovulation., Conclusion: Follicular growth rate, degree of uterine oedema and the number of antral follicles are higher in young mares with high AFC. However, the rate of embryonic recovery is not influenced by donors' age or AFC., Competing Interests: Competing interests: None declared., (© British Veterinary Association 2020. No commercial re-use. See rights and permissions. Published by BMJ.)