Multiple Ovulation Research Articles

Roe Deer Reproduction in Western Poland: The Late Autumn Rut Phenomenon.

Roe deer (Capreolus capreolus L.) populations in Poland are characterized by low productivity, which is why their reproductive potential was investigated. The presence of corpora lutea (CL) on the ovaries of females hunted in autumn and winter in the years 2015 and 2016 was assessed. Most animals were post-rut and most often had multiple ovulations. However, in early autumn 2015, 60% of the females had no CL. Therefore, they did not participate in mating at the turn of July/August. Those that did ovulate were found to have one CL. In late autumn, 97% of the females were post-rut, mostly with twin ovulations. This confirmed the occurrence of a late autumn rut. This phenomenon had been suggested in studies but not confirmed. In contrast, almost 100% of roe deer ovulated in summer in 2016. It was hypothesized that the reason for the low mating activity of roe deer in the summer of 2015 was heat stress and limited food resources. The summer was exceptionally hot, with many days in July and August when temperatures exceeded 30 °C. The heat combined with low rainfall led to extreme drought. Meanwhile, July and August are the months of mating activity for roe deer. The late autumn rut allowed the roe deer-a monoestrous species-to limit the consequences of a decrease in mating activity or fertility during the hot summer. Global warming may affect roe deer reproduction, so climatic conditions should be considered in population studies, not only in terms of food availability.

Effect of pre-treatment with deslorelin on the ovarian response of ewes superovulated with FSH.

This study evaluated the use of the GnRH agonist hormone, deslorelin, to control the follicular population before initiating multiple ovulation and embryo transfer (MOET) treatment. Twenty-four cross-bred Santa Inês ewes, aged between 2 and 4 years, were randomly assigned to either a control group (n = 11) or a treated group (n = 13). All ewes received an intravaginal device containing 60 mg of medroxyprogesterone acetate on day 0, and a new device on day 7, which remained in place until day 14. Additionally, the ewes were administered 125 μg of cloprostenol on day 7. The superovulatory treatment involved administering 200 mg of pFSH, divided into eight decreasing doses at 12-h intervals starting on day 12. On day 14, 300 IU of eCG was administered. In the deslorelin group, three doses of 100 μg of deslorelin were administered starting on day 3 after the insertion of the vaginal device, with subsequent doses given at 72-h and 144-h intervals. Natural mating was performed 36 h after the removal of the progesterone implant using males with proven fertility. Embryo collection took place on the 6th day after mating, and the recovered structures were quantified and evaluated for quality and developmental stage. Transrectal ultrasonography was conducted on days 12, 16 and 21 to evaluate the ovaries, specifically to assess the ovarian follicular population and the presence of the corpus luteum. Ewes in the control group had higher embryo recovery rates (p < .01) compared to the treated group (5.2 ± 0.8 vs. 1.1 ± 0.8), with differences observed primarily in the number of morulae. The number of corpus luteum observed during the laparotomy on day 21 was significantly higher (p < .01) in the control group (10.44 vs. 4.5 corpus luteum per ewe). Yet, the treated group had a significantly higher number of follicles (p < .05) on the first day of pFSH application (5.5 vs. 3.0 follicles per ewe). In conclusion, although the inclusion of deslorelin in the superovulation protocol resulted in increased synchronization of oestrus and follicle number, it did not lead to an increase in the number of corpus luteum or harvested embryos.

A retrospective analysis of conception per embryo transfer in dairy cattle in South Korea

The bovine embryo production industry has seen significant growth over the past two decades, particularly in the production of in vitro produced embryos. This growth, driven by advancements in cryopreservation, in vitro culture mediums, ovum pick-up (OPU) procedures, ultrasonography devices, and embryo transfer (ET) has been notable. Particularly, ET is crucial for disseminating high genetic merit and amplifying foreign breeds by importing frozen embryos. This retrospective study aimed to assess factors affecting conception per embryo transfer (CPET) in Holstein-Friesian cattle in South Korea from October 2008 to July 2022. We evaluated type of embryo breed, type of embryo production (fresh and frozen; in vitro and in vivo production), recipient conditions including estrus type, corpus luteum quality, parity (nulliparous heifers, primiparous, and multiparous cows), and the daily mean temperature-humidity index (THI) as an index for heat stress. Type of embryo breed and estrus had no significant impact on CPET. However, we observed higher CPET in recipients with good quality corpus luteum, nulliparous heifers, and surrogates receiving fresh in vitro and frozen in vivo embryos. Importantly, CPET was not adversely affected by mild heat stress conditions (up to daily mean THI 76), indicating that using frozen in vivo embryos produced by multiple ovulation embryo transfer and fresh in vitro embryos by OPU-ET can help alleviate the subfertility issues in dairy cattle caused by global warming in Korea.

O-062 New strategies in ovarian stimulation

Abstract Controlled Ovarian stimulation (COH) has an objective to induce multiple ovulations in order to yield a multiple oocyte harvest and offer multiple embryos available for transfer thereby increasing the efficacy of ART. The theory of a multicyclic development of follicles during the menstrual cycle has prompted new approaches to ovarian stimulation. Research has suggested that recruitable antral follicles are continuously present in the ovaries during the menstrual cycle, and ultrasonographic studies have demonstrated that multiple cohorts or ‘waves’ of 2–5 mm follicles are recruited continuously during a menstrual cycle. Because of the asynchrony between endometrial receptivity and embryo development with a ‘non-conventional start’ stimulation, all the embryos are generally cryopreserved and transferred subsequently. This deferred transfer policy is currently possible given the advances in vitrification techniques, with success rates comparable to those following transfer with ‘fresh’ embryos. New stimulation approaches, together with advanced cryopreservation techniques, allow for a total ‘disarticulation’ between the time of the menstrual cycle, ovarian stimulation start and embryo transfer. This new approach to ovarian stimulation is particularly useful for women seeking fertility preservation, especially where a shortened time to starting cancer treatment is desirable. Also, poor responders could benefit from the new stimulation protocols by continuing ovarian stimulation after the first oocyte retrieval, thereby obtaining more oocytes or embryos compared with the conventional approach. In the quest to reduce the incidence of OHSS and make the clinic OHSS free various antagonist protocols are now routinely used. These and other new protocols discussed will include Late initiation of stimulation, long acting antagonist. Individualised (iCOS) to include mild stimulation, random start, Dual stimulation and double trigger are done to improve efficacy and success rates in certain group of patients. New drugs like Long acting FSH and oral antagonist with their results will be presented. Pre-Treatment medications like Estrogens, Progestogens, Combined CoC, antagonist will also be discussed.

P-519 Polygenic scores for DZ twinning in mothers of naturally and ART conceived twins and singletons

Abstract Study question Does the polygenetic score (PGS) of dizygotic (DZ) twinning differ among mothers of naturally conceived DZ twins and mothers of ART twins, or singleton mothers? Summary answer Mothers of naturally conceived DZ twins have the highest PGS for DZ twinning, while mothers who received hormonal induction of ovulation have the lowest PGS. What is known already Natural DZ twinning results from a double ovulation and has a genetic component. Endocrinology studies in mothers of DZ twins observed higher levels of follicle-stimulating hormone (FSH). FSHB and SMAD3 were the first loci identified as significant in genome-wide association studies (GWAS) of having DZ twins. The GWAS also showed a genetic correlation with anovulatory infertility and a PGS that explained ∼1.5% of the phenotypic variance when comparing mothers of naturally conceived DZ twins to controls. As DZ twinning has been proposed to be a measure of female fertility, we hypothesise that a low PGS could indicate infertility. Study design, size, duration We compare the DZ twinning PGS in mothers of naturally conceived DZ twins with other mothers in the Netherlands twin register (NTR) and the Norwegian Mother, Father and Child Cohort Study (MoBa). We tested if the PGS is lower in infertility groups (ART use). We included 1,677 cases, 2,828 other twin mothers and 2,686 controls from the NTR and 396 cases, 1,285 twin mothers and 74,613 singleton mothers from MoBa. Participants/materials, setting, methods We calculated PGSs of DZ twinning for all participants employing LDPred0.9 based on the summary statistics from the 2024 GWAS (Mbarek et al., 2024). Differences between groups were assessed by logistic regression, with corrections for relatedness, maternal age, maternal age at first birth, BMI, height, parity and technical covariates. Main results and the role of chance Mothers of naturally conceived DZ twins differed in age, body composition, parity and smoking behaviour compared to the other groups. The PGSs were higher in mothers of naturally conceived DZ twins (N = 1,677) compared to all other twin mothers, including mothers of naturally conceived MZ twins (N = 2,165) and mothers who conceived MZ (N = 64) or DZ twins through ART (N = 559) and compared to controls (N = 2,686; p &amp;lt; 0.01). For ART mothers whose fertility treatment involved hormones without IVF (N = 257), the PGS was significantly lower than for the other three groups (DZ-IVF and MZ mothers). In MoBA, we replicate our findings of the higher PGSs for mothers who naturally conceived their DZ twin pregnancy (N = 705) compared to mothers of naturally conceived MZ twins (N = 313) and mothers who conceived their DZ twins through ATR (N = 328) (p &amp;lt; 0.05). However, we do not observe differences between the mothers of naturally conceived DZ twins and the mothers of ART MZ twins in MoBa. We are currently adding analysis comparing the mothers of naturally conceived DZ with mothers who naturally or through ART conceived singletons. Limitations, reasons for caution In interpreting our results we should consider that the MZ ART groups are relatively small. Also, we did not have information on the reasons for ART use so we could not distinguish between maternal, paternal or couple fertility issues. Wider implications of the findings Mothers of naturally conceived DZ twins are genetically different from other mothers. Genetic influences found for DZ twinning mainly affect multiple ovulation, rather than other characteristics required for a twin pregnancy. The DZ twinning PGS may be associated with infertility. Trial registration number not applicable

Production, cryopreservation, and transfer of Murrah embryos through Multiple Ovulation and Embryo Transfer (MOET) technology in Sri Lanka

Production, cryopreservation, and transfer of Murrah embryos through Multiple Ovulation and Embryo Transfer (MOET) technology in Sri Lanka

Development of a biotechnology process for the production of a novel hyperglycosylated long-acting recombinant bovine follicle-stimulating hormone.

Follicle-stimulating hormone (FSH) is an important protein used for bovine ovarian hyperstimulation in multiple ovulation and embryo transfer technology (MOET). Several attempts to produce bovine FSH (bFSH) in recombinant systems have been reported, nonetheless, up to date, the most commonly used products are partially purified preparations derived from porcine or ovine (pFSH or oFSH) pituitaries. Here we describe the development of a biotechnology process to produce a novel, hyperglycosylated, long-acting recombinant bFSH (LA-rbFSH) by fusing copies of a highly O-glycosylated peptide. LA-rbFSH and a nonmodified version (rbFSH) were produced in suspension CHO cell cultures and purified by IMAC with high purity levels (>99%). LA-rbFSH presented a higher glycosylation degree and sialic acid content than rbFSH. It also demonstrated a notable improvement in pharmacokinetic properties after administration to rats, including a higher concentration in plasma and a significant (seven-fold) reduction in apparent clearance (CLapp). In addition, the in vivo specific bioactivity of LA-rbFSH in rats was 2.4-fold higher compared to rbFSH. These results postulate this new molecule as an attractive substitute for commercially available porcine pituitary-derived products.

Development and characterization of a novel variant of long-acting bovine follicle-stimulating hormone (brscFSH)

Assisted reproduction is a key aspect of modern animal breeding, providing valuable assistance in improving breeding programs. In this field, the administration of exogenous hormones, such as follicle-stimulating hormone (FSH), plays a crucial role in the induction of multiple ovulations. However, commercial FSH used in veterinary practice has been derived primarily from pituitary glands, obtained mostly from pigs for nearly four decades. Although these hormones have contributed significantly to the advancement of assisted reproductive techniques, they have certain limitations that warrant further improvements. These limitations include contamination with luteinizing hormone (LH), the potential risk of pathogen contamination, the potential to trigger an immune response in non-pig species, and the short half-life in circulation, requiring the implementation of complex 8-dose superovulation schedules. Our research team has developed and characterized a new variant of bovine follicle-stimulating hormone (bscrFSH) to address these limitations. The new hormone is produced recombinantly in CHO cell cultures, with a specific productivity of about 30 pg/cell/day. The bscrFSH can be purified to a high purity of 97 % using a single step of immobilized metal affinity chromatography (IMAC). N-glycan analysis of bscrFSH showed that approximately 74 % of the glycans corresponded to charged structures, including mono-, di-, tri-, and tetra-sialylated glycans. Superovulation trials conducted in cattle revealed that bscrFSH, administered at a total dose of about 0.5 μg per kg of body weight, using a decrescent schedule of 4 doses with 24-h intervals, resulted in an average yield of 8–12 transferable embryos per animal. Further research is required; however, the preliminary findings indicate that bscrFSH, currently packaged under the provisional brand name of Cebitropin B, holds potential as a commercial product for assisted reproduction in ruminants.

Innovative Reproductive Technology in Animal Breeding: A Review

The profitability and the intensity achieved by the genetic improvement is determined by the reproductive performance of the farm animals. The major cause of economic loss is reproductive inefficiency in the livestock sector. In the past few years, several advances have been made in reproductive technologies (RTs) to increase the efficiency of genetic improvement. These technologies include Artificial Insemination, Sexed semen Technology, Cryopreservation, Multiple Ovulation and Embryo Transfer, In-vitro Embryo Production (IVEP), Transgenesis etc. The problems of infertility of male or female, postpartum infertility in high-yielding animals of high genetic merit individuals and breeding of distant species has been overcome through these technologies. The inclusion of these techniques in animal breeding had made possible to obtain large number of progenies from genetically superior animals. This review, discuss the various innovative reproductive technologies implemented in livestock sector.

Machine-learning models evaluate embryo morphokinetics to detect subclinical signs of heat stress in embryos produced by multiple ovulation embryo transfer

Increased demand for animal-based protein, driven by the in­creasing world population, has emphasized the use of bovine embryo transfer (ET). However, this demand is predominantly in locations where environmental conditions, like heat stress, are inhibiting reproductive performance. Heat stressed do­nors have decreased viable blastocyst production, greater vari­ability in embryonic gene expression and lower ET pregnancy outcomes in current literature. These embryonic changes are nonidentifiable to the human eye, highlighting the need for solutions to identify compromised embryos and improve ET efficiencies. Machine-learning artificial intelligence has been used to evaluate health by assessing embryo morphokinetics, or time-specific morphological changes during embryo devel­opment, from videos of embryos in standard media. Thus, the objective was to use machine learning to detect real-time mor­phokinetic activity of embryos from donor cows undergoing multiple ovulation embryo transfer (MOET) based on seasonal exposure to heat stress.

Twin management in the mare: A review.

Twin gestation in the mare is undesirable and can have disastrous consequences. As in many cases, the key to success in twin management lies in a thorough follow-up and accurate recording of clinical findings in the pre-breeding examination. A pregnancy diagnosis in the mobility phase is imperative for a good outcome in the event of twin reduction. If a twin gestation is not diagnosed during this early pregnancy stage, several other procedures exist for managing post-fixation twins (>16 days) with varying degrees of success. Most twin pregnancies are the result of multiple ovulations (dizygotic twins). However, monozygotic twins are also sporadically diagnosed, due to the increasing number of transferred in vitro produced equine embryos. In these cases, the most optimal treatment strategy still needs to be determined. This review provides an overview of the various twin reduction techniques described with the expected prognosis as well as of some less reported techniques with their results. In addition, physiological events and the reduction techniques are demonstrated to the user in virtual 3-dimensional illustrations.

Clinical and experimental substantiation of the use of reduced concentrations of FSH during superovulation in highly productive cows

The purpose of the studies was to substantiate the use of reduced concentrations of follicle-stimulating hormone to induce superovulation in highly productive cows and heifers. The research was carried out in one of the farms in the Leningrad region. After analyzing breeding cards and based on ultrasound visualization of the reproductive organs, promising animal embryo donors were selected - cows after the first calving and heifers that had reached physiological maturity, Holstein breed. According to the principle of analogues, groups of experimental animals Ⅰ and Ⅱ were formed, which included cows, and groups Ⅲ and Ⅳ, which included heifers, eight and six animals each, respectively. To induce multiple ovulation, hormonal treatment regimens were used, presented in Table 1. Hormonal treatment regimens were compiled in accordance with methodological recommendations on the procedure and conditions for applying the method of cattle embryo transplantation, as well as in accordance with the results of previously conducted studies and analysis of literary sources. As a result of the studies, it was established that all experimental animals responded equally to the hormonal regimen recommended by the manufacturer of exogenous gonadotropins and the experimental regimen, with a reduced concentration of the latter. It was revealed that there were no significant differences in the number of corpora lutea in the groups of cows and heifers when using a standard scheme for stimulating polyovulation and with a reduced content of follicle-stimulating hormone. There was also no difference between the experimental groups of cows and heifers in the number of embryos obtained. Analyzing the data obtained, we can conclude that it is advisable to use reduced concentrations of pituitary gonadotropins to stimulate multiple ovulation in highly productive cows and heifers.

A case report of a family with developmental arrest of human prokaryotic stage zygote.

To study the genetic variation leading to the arrest phenotype of pronuclear (PN) zygotes. We recruited a family characterized by recurrent PN arrest during in vitro fertilization (IVF) and intracytoplasmic sperm injection cycles (ICSI) and performed whole-exome sequencing for 2 individuals. The transcriptome profiles of PN-arrest zygotes were assessed by single-cell RNA sequencing analysis. The variants were then validated by PCR amplification and Sanger sequencing in the affected individuals and other family members. A family characterized by recurrent PN arrest during IVF and ICSI cycles were enrolled after giving written informed consent. Peripheral blood samples were taken for DNA extraction. Three PN-arrest zygotes from patient III-3 were used for single-cell RNA-seq as described. This phenotype was reproduced after multiple cycles of egg retrieval and after trying different fertilization methods and multiple ovulation regimens. The mutant genes of whole exon sequencing were screened and verified. The missense variant c. C1630T (p.R544W) in RGS12 was responsible for a phenotype characterized by paternal transmission. RGS12 controls Ca2+ oscillation, which is required for oocyte activation after fertilization. Single-cell transcriptome profiling of PN-arrest zygotes revealed defective established translation, RNA processing and cell cycle, which explained the failure of complete oocyte activation. Furthermore, we identified proximal genes involved in Ca2+ oscillation-cytostatic factor-anaphase-promoting complex (Ca2+ oscillation-CSF-APC) signaling, including upregulated CaMKII, ORAI1, CDC20, and CDH1 and downregulated EMI1 and BUB3. The findings indicate abnormal spontaneous Ca2+ oscillations leading to oocytes with prolonged low CSF level and high APC level, which resulted in defective nuclear envelope breakdown and DNA replication. We have identified an RGS12 variant as the potential cause of female infertility characterized by arrest at the PN stage during multiple IVF and ICSI.

Effects of donor age and reproductive history on developmental potential of ovum pickup oocytes in Japanese Black cattle (Wagyu)

The objectives of this study were to analyze the (1) effects of donor age and multiparity on development of in vitro fertilization (IVF) embryos after ovum pickup (OPU), (2) effects of repeated and consecutive OPU-IVF procedures on embryo development, and (3) embryo production from OPU-IVF in donors with differing embryo yields after multiple ovulation and embryo transfer technology (MOET) in Japanese Black cattle (Wagyu). Donors were pre-treated with low-dosage follicle-stimulating hormone (FSH; 200 IU total), and oocytes were collected via OPU and fertilized by IVF to generate blastocysts. The number of oocytes collected per OPU session per donor was lower in heifers (2–4 years old, 5.3 oocytes) than in primiparous and pluriparous cows (2–10 years old, 13.6–19.1 oocytes; P < 0.05). Rates of blastocyst development for oocytes from heifers (33.1%) were lower than for those from cows (2–10 years old, 44.1–54.3%; P < 0.05), and average blastocyst yield/OPU/animal was lower in heifers (3.7) than in 5–6 years old cows (10.1; P < 0.05). Donors undergoing five consecutive OPU-IVF sessions after low-dosage FSH showed similar oocyte retrieval (12.2–15.1 oocytes per OPU/animal), blastocyst development rates (35.6–45.0%), and embryo yield/OPU/animal (4.8–5.8; P > 0.05) across sessions. Additionally, embryo yield from OPU-IVF was significantly improved in animals with previous low embryo yield from MOET (5.9 vs. 2.6, respectively, P < 0.05). These results indicate that Wagyu cows with previous births can be more productive as OPU-IVF donors than heifers, and oocytes from donors undergoing to five consecutive OPU-IVF cycles are competent for embryo development without loss of embryo yield/OPU/animal. Moreover, OPU-IVF can be used for embryo production and breeding from all elite Japanese Black cattle, regardless of previous low embryo yield in routine MOET.

Factors influencing ovum pick-up technique results in cattle

The Ovum Pick-Up (OPU) technique, which is used in in vitro embryo production (IVP) to retrieve immature oocytes from live donor animals, is one of the most important biotechnological procedures used in cattle breeding. The most important advantage of this technology is that it allows for the reproducible retrieval of immature oocytes from living donor animals. It is particularly useful in dairy cattle breeding to address infertility issues and boost the production of superior animals with high genetic value. The OPU technique offers several advantages, including its applicability to cows ranging from six-month-old calves to the first three months of pregnancy, its effectiveness in animals with genital tract infection or acyclic cattle, and its ability to yield a higher number of embryos within the same period compared to the Multiple Ovulation and Embryo Transfer (MOET) technique. Understanding and improving the technical and biological factors influencing the OPU procedure is necessary to increase and optimize donor animal use in IVP and the number of quality oocytes obtained. This review aims to examine the specifics of the OPU approach and the factors influencing its performance in light of contemporary literature, as well as to propose fresh ideas to researchers.

Angiosperm-wide analysis of fruit and ovary evolution aided by a new nuclear phylogeny supports association of the same ovary type with both dry and fleshy fruits.

Fruit functions in seed protection and dispersal and belongs to many dry and fleshy types, yet their evolutionary pattern remains unclear in part due to uncertainties in the phylogenetic relationships among several orders and families. Thus we used nuclear genes of 502 angiosperm species representing 231 families to reconstruct a well supported phylogeny, with resolved relationships for orders and families with previously uncertain placements. Using this phylogeny as a framework, molecular dating supports a Triassic origin of the crown angiosperms, followed by the emergence of most orders in the Jurassic and Cretaceous and their rise to ecological dominance during the Cretaceous Terrestrial Revolution. The robust phylogeny allowed an examination of the evolutionary pattern of fruit and ovary types, revealing a trend of parallel carpel fusions during early diversifications in eudicots, monocots, and magnoliids. Moreover, taxa in the same order or family with the same ovary type can develop either dry or fleshy fruits with strong correlations between specific types of dry and fleshy fruits; such associations of ovary, dry and fleshy fruits define several ovary-fruit "modules" each found in multiple families. One of the frequent modules has an ovary containing multiple ovules, capsules and berries, and another with an ovary having one or two ovules, achenes (or other single-seeded dry fruits) and drupes. This new perspective of relationships among fruit types highlights the closeness of specific dry and fleshy fruit types, such as capsule and berry, that develop from the same ovary type and belong to the same module relative to dry and fleshy fruits of other modules (such as achenes and drupes). Further analyses of gene families containing known genes for ovary and fruit development identified phylogenetic nodes with multiple gene duplications, supporting a possible role of whole-genome duplications, in combination with climate changes and animal behaviors, in angiosperm fruit and ovary diversification.

Negative photoperiod induces an increase in the number of ovulations in dairy cattle.

This study sought to examine the impact of negative photoperiod on the incidence of multiple ovulations and pregnancies in dairy cattle. The study population consisted of 5,373 pregnant cows in their third or greater lactation that experienced their first post-partum pregnancy after spontaneous estrus. The positive photoperiod (increasing day-length) extends from December 22 to June 21, whereas the negative photoperiod (decreasing day-length) extends from June 22 to December 21. The odds ratios (ORs) for multiple ovulations and pregnancies in cows that became pregnant during the negative photoperiod and the remaining cows that became pregnant during the positive photoperiod were 1.4 and 1.3 (P < 0.0001), respectively. The ORs for cows that became pregnant ≥ 90 days in milk and the remaining cows that became pregnant < 90 days in milk were 4.3 and 4.1 (P < 0.0001), respectively. No significant differences were detected in the monthly rates of multiple ovulations or pregnancies during positive and negative photoperiods. Thus, the present study demonstrates that the ovarian function in cows is related to changes in day-length, with decreasing day-length being associated with greater multiple ovulation and pregnancy rates. The present study also shows that positive and negative photoperiods exhibit different trends. The results of this study are consistent with a growing body of work demonstrating the effects of photoperiod patterns on the reproductive physiology of cows, with clear implications for twin pregnancy prevention.

Bridging the Gap: Animal Models in Next-Generation Reproductive Technologies for Male Fertility Preservation.

This review aims to explore advanced reproductive technologies for male fertility preservation, underscoring the essential role that animal models have played in shaping these techniques through historical contexts and into modern applications. Rising infertility concerns have become more prevalent in human populations recently. The surge in male fertility issues has prompted advanced reproductive technologies, with animal models playing a pivotal role in their evolution. Historically, animal models have aided our understanding in the field, from early reproductive basic research to developing techniques like artificial insemination, multiple ovulation, and in vitro fertilization. The contemporary landscape of male fertility preservation encompasses techniques such as sperm cryopreservation, testicular sperm extraction, and intracytoplasmic sperm injection, among others. The relevance of animal models will undoubtedly bridge the gap between traditional methods and revolutionary next-generation reproductive techniques, fortifying our collective efforts in enhancing male fertility preservation strategies. While we possess extensive knowledge about spermatogenesis and its regulation, largely thanks to insights from animal models that paved the way for human infertility treatments, a pressing need remains to further understand specific infertility issues unique to humans. The primary aim of this review is to provide a comprehensive analysis of how animal models have influenced the development and refinement of advanced reproductive technologies for male fertility preservation, and to assess their future potential in bridging the gap between current practices and cutting-edge fertility techniques, particularly in addressing unique human male factor infertility.

Proteomic Analysis Identifies Distinct Protein Patterns for High Ovulation in FecB Mutant Small Tail Han Sheep Granulosa Cells.

The Booroola fecundity (FecB) mutation in the bone morphogenetic protein receptor type 1B (BMPR1B) gene increases ovulation in sheep. However, its effect on follicular maturation is not fully understood. Therefore, we collected granulosa cells (GCs) at a critical stage of follicle maturation from nine wild-type (WW), nine heterozygous FecB mutant (WB), and nine homozygous FecB mutant (BB) Small Tail Han sheep. The GCs of three ewes were selected at random from each genotype and consolidated into a single group, yielding a total of nine groups (three groups per genotype) for proteomic analysis. The tandem mass tag technique was utilized to ascertain the specific proteins linked to multiple ovulation in the various FecB genotypes. Using a general linear model, we identified 199 proteins significantly affected by the FecB mutation with the LIMMA package (p < 0.05). The differential abundance of proteins was enriched in pathways related to cholesterol metabolism, carbohydrate metabolism, amino acid biosynthesis, and glutathione metabolism. These pathways are involved in important processes for GC-regulated 'conservation' of oocyte maturation. Further, the sparse partial least-squares discriminant analysis and the Fuzzy-C-mean clustering method were combined to estimate weights and cluster differential abundance proteins according to ovulation to screen important ovulation-related proteins. Among them, ZP2 and ZP3 were found to be enriched in the cellular component catalog term "egg coat", as well as some apolipoproteins, such as APOA1, APOA2, and APOA4, enriched in several Gene Ontology terms related to cholesterol metabolism and lipoprotein transport. A higher abundance of these essential proteins for oocyte maturation was observed in BB and WB genotypes compared with WW ewes. These proteins had a high weight in the model for discriminating sheep with different FecB genotypes. These findings provide new insight that the FecB mutant in GCs improves nutrient metabolism, leading to better oocyte maturation by altering the abundance of important proteins (ZP2, ZP3, and APOA1) in favor of increased ovulation or better oocyte quality.

Evaluation of a long-acting recombinant bovine FSH for multiple ovulation and embryo transfer in dromedary camels

The present study was conducted to test a new super-agonist recombinant bovine FSH (rbFSH) to induce superovulation (SOV) in dromedary camels. In experiment I, a single IM injection of 40, 60, 80, 100, or 120 µg rbFSH was administered (4 donors per group) to determine the effective dose resulting in acceptable multiple ovulation and embryo yield. Administration of 40 µg was ineffective, while 100 and 120 µg were associated with increased numbers of developed follicles, corpora lutea, and recovered embryos compared to administration of 60 and 80 µg. In experiment II, donors were divided into treatment groups to compare rbFSH with two conventional protocols for SOV. Donors received a single dose of 2000 IU eCG in combination with 400 mg porcine follicle-stimulating hormone (pFSH; Folltropin-V®; Group 1, n = 29) or 500 µg of pFSH with 100 µg of pLH (Stimufol®; Group 2, n = 16). Group 3 (n = 19) received a single dose of 100 µg rbFSH. No difference was found in the size and number of follicles per donor. Response time, ovulation rate, and the number of corpora lutea and recovered embryos per donor were similar in all groups. The number of medium-sized and transparent embryos decreased while the number of small-sized and semi-transparent embryos increased in Group 3 (rbFSH) compared to the other two groups. The pregnancy rate of the recipients at 10 days post-ET, at two months of gestation, and the rate of early pregnancy loss (EPL) did not differ among the groups. In conclusion, a single IM administration of 100 µg rbFSH induces a successful superovulation in dromedary camels and has the advantage of reducing stress associated with multiple FSH administration of the conventional protocols.