Ovum Pick-up Procedure Research Articles

Use of excised ovaries for oocyte recovery by ultrasound guided follicular aspiration – Validation of an experimental model for research purposes in live mares ovum pick up

Ovum pick-up (OPU) by transvaginal ultrasound guided follicle aspiration in mares is a common assisted reproductive technique used for oocyte recovery and in vitro production of horse embryos. There has been relatively little research into the factors influencing oocyte recovery in OPU from live mares. The objective of this study was to compare oocyte recovery and morphology of ultrasound-guided follicle puncture and aspiration in live mares and in postmortem excised ovaries, in order to validate an experimental model for research purposes of the efficiency of OPU in mares. Data from OPU performed in 12 mares from a commercial program (follicle numbers, oocyte recovery and oocyte morphology) were compared to that obtained from ultrasound-guided follicle puncture of 13 postmortem excised ovaries from slaughtered mares processed within 2 h of slaughter. In both groups, the OPU was performed by the same operator using the same equipment and OPU technique. The recovered oocytes per aspirated follicle was higher (P < 0.05) in the postmortem group (105/166, 63.2 %) than in live mares (138/261, 52.9 %). There was more (P < 0.05) expanded cumulus oocyte complexes in the postmortem than in the live mares (18 % vs. 2.9 %). Several oocytes (5 oocytes from 81 aspirated follicles) were found in the leaked fluid which overflowed during follicle flushing of postmortem ovaries. In conclusion, the higher recovery rate obtained in the excised ovaries and the finding of oocytes in the leaked fluid during OPU, suggests that there is still room for improvement in the in vivo OPU technique. Utilizing postmortem excised ovaries could offer an alternative for further research into factors affecting oocyte recovery and oocyte leakage during OPU procedures.

The Role of Total Oxidant and Antioxidant Levels in Follicular Fluid in Unexplained İnfertility

Background: Unexplained infertility is defined as the absence of any pathology in the basic evaluation performed in couples who cannot achieve pregnancy after 1 year of unprotected sexual intercourse. The results of tests examining the causes of infertility show no identifiable cause in almost 15% of couples. Aim: The aim of this study was to investigate the effects of reactive oxygen species (ROS) on pregnancy and embryos. Methods: This study included 200 patients, aged between 20–44 years, with unexplained infertility, who had recurrent intrauterine inseminations failures and hence started in vitro fertilization (IVF)/intracytoplasmic sperm injection treatment. Some amounts of waste follicular fluid samples were collected by embryologists from the oocytes of these patients during the ovum pick-up procedure. Next, total antioxidant status (TAS), total oxidant status (TOS), and oxidative stress index (OSI) values were calculated in the biochemistry laboratory. Results: In terms of pregnancy status, both follicular TOS and OSI values were not significantly different in patients with biochemical and clinical pregnancy, whereas TAS values were significantly higher in patients with pregnancy (P &lt; 0.05). In terms of embryo quality, no significant difference was observed in TAS, TOS, and OSI values between grade 1 and 2 embryos, whereas pregnancy rates were significantly higher in patients who received grade 1 embryo transfer (P &lt; 0.05). However, the follicular fluid TAS levels were significantly lower in smoking patients than in those who did not smoke; TOS and OSI levels were significantly higher. Conclusion: This study showed that exposure to oxidative stress might be a causative factor for infertility. In addition, ROS decreased the level of TAS by increasing OSI in the follicular fluid; thus, antioxidant supplementation might be a necessity.

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.

Efficiency of equine oocyte recovery from post mortem ovaries or by ovum pick up

Most oocytes used for cloned embryo production are obtained from abattoir ovaries even though in some countries, slaughtering horses is not allowed. The advantage of scraping post mortem ovaries is the high number of oocytes available in a short period of time to perform somatic cell nuclear transfer. However, mare age, reproductive and health status or ovary manipulation by abattoir staff are not controlled before slaughter and the effect on oocyte quality and early embryo development is uncertain. Ovum pick up (OPU) is an alternative technique to obtain oocytes ex vivo from controlled mares and embryos obtained from those oocytes potentially have better development. Cortez et al. (Journal of Equine Veterinary Science 113 2022; 14) reported higher cleavage and blastocyst rates in cloned embryos from OPU-derived rather than post mortem derived ovaries. The aim of this retrospective study was to determine and compare the oocyte recovery rates from abattoir-sourced ovaries or collected by OPU. Data was collected between September and November 2022 in Argentina. For statistical analysis, generalized linear models were used. A total of 3476 ovaries were collected from the abattoir and 16% (557/3476) were discarded since antral follicles were not visible. Using the scraping technique, 8392 oocytes were recovered in 24 sessions obtaining an averageof 350 oocytes per session. Mean (± SD) oocyte recovered per scraped ovary was 2.9±0.4 and for all collected ovaries per session was 2.4±0.3. On average the scraping time calculated per mare (pair of ovaries) for all collected ovaries per session was 1 min and 36 sec (59 sec-2 min). OPU was performed in 44 mixed breed healthy mares during the southern hemisphere breeding season in which a mean of 16.7 (7-29) follicles were aspirated per mare in an average of 27 min and 7 sec (6 min and 3 sec- 58 min). Oocyte recovery rate per aspirated follicle was 63.3% and a mean of 9.7 oocytes were retrieved per OPU. Mean oocyte retrieval per mare was higher using OPU than scraping (9.7 ± 4.8 vs. 5.8 ± 0.9; p<0.0001). Although oocyte recovery per follicle could not be compared in this study these results could be influenced by mares’ selection for OPU although there was not a minimum number of follicles set as a criteria to perform OPU. The data show that the OPU procedure is a suitable technique to obtain oocytes for equine cloning programs. In some circumstances, the procedures to obtain oocytes by OPU are more expensive than those recovered by scraping, however published data suggest that fewer oocytes recovered by OPU are needed to obtain a cloned embryo.

The relationship between bovine blastocyst formation in vitro and follicular fluid amino acids

Follicular fluid has been found as a possible source of metabolic predictors for oocyte competence, and it is conveniently accessible during ovum pick-up (OPU). We used the OPU procedure to recover oocytes from 41 Holstein heifers for in vitro embryo production in this study. Follicular fluid was collected during OPU in order to establish a link between follicular amino acids and blastocyst formation. Each heifer's oocytes were collected, matured in vitro for 24 h and fertilized separately. The heifers were then divided into two groups based on blastocyst formation: those that produced at least one blastocyst (the blastocyst group, n = 29) and those that did not (the failed group, n = 12). The blastocyst group had higher follicular glutamine concentrations and lower aspartate levels than the failed group. Furthermore, network and Spearman correlation analyses revealed a link between blastocyst formation and aspartate (r = −0.37, p = 0.02) or glutamine (r = 0.38, p = 0.02). The receiver operator characteristic curve revealed that glutamine (AUC = 0.75) was the greatest predictor of blastocyst formation. These findings revealed that follicular amino acid levels in bovines can be used to predict blastocyst development.

Ethical Analysis of the Application of Assisted Reproduction Technologies in Biodiversity Conservation and the Case of White Rhinoceros (Ceratotherium simum) Ovum Pick-Up Procedures.

Originally applied on domestic and lab animals, assisted reproduction technologies (ARTs) have also found application in conservation breeding programs, where they can make the genetic management of populations more efficient, and increase the number of individuals per generation. However, their application in wildlife conservation opens up new ethical scenarios that have not yet been fully explored. This study presents a frame for the ethical analysis of the application of ART procedures in conservation based on the Ethical Matrix (EM), and discusses a specific case study—ovum pick-up (OPU) procedures performed in the current conservation efforts for the northern white rhinoceros (Ceratotherium simum cottoni)—providing a template for the assessment of ART procedures in projects involving other endangered species.

Association of follicular fluid parameters at the time of ovum pick-up with in vitro blastocyst formation in Holstein heifers: a multi-level analysis

Pooled follicular fluid is easily accessible during ovum pick-up (OPU) and can be a potential source of non-invasive biochemical predictors for oocyte competence. Using the OPU procedure, oocytes recovered from a total of 60 Holstein heifers were used in in vitro embryo production. Pooled follicular fluid was collected at the time of OPU in order to establish a relationship between follicular biochemical components and the formation of blastocyst. The oocytes from each heifer were pooled together, matured in in vitro maturation medium for 24 h and then fertilized with sperm cells. The resulting zygotes were cultured for 7 days and the formation of blastocyst was recorded. Next, heifers were classified on the basis of the blastocyst formation, cleaved heifers and failed heifers. Follicular glucose concentrations were higher and the amount of C-reactive protein (CRP) was lower in cleaved heifers than in failed heifers. Moreover, the receiver operator characteristic (ROC) curve, clustering and network analysis showed the closeness between CRP, total bilirubin, and blastocyst yield. In conclusion, the findings imply that CRP follicular concentrations, which have been linked to heifer blastocyst formation, should be investigated further.

INTRAOVARIAN INJECTION OF ALLOGENEIC MESENCHYMAL STEM CELLS: FEASIBILITY AND SAFETY ASSESSMENT IN BOVINES

<h3>Background</h3> Ovum pick-up (OPU) is a technique widely used for production of bovine embryos in-vitro. However, continuous OPU can be associated with the appearance of often irreversible ovarian lesions. The possibility of using stem cell therapy for treatment of infertility both in animals and in humans is currently under investigation, but the safety of intraovarian infusion of stem cells in bovines has not been established at this point. Therefore, in the present study we determined the feasibility and safety of intraovarian application of allogeneic MSC derived from adipose tissue of healthy cows. <h3>Methods</h3> Two OPU procedures were executed 15 days apart. Immediately following the second OPU, 2.5 million allogeneic MSCs were injected to the ovarian cortex of healthy nelore (n=5) and girolando (N=5) cows. The animals were subsequently evaluated by ultrasonography and clinical examination at 24, 48 and 72 hs post MSC injection. At 15, 30 and 45 days post MSC injection, new OPU procedures were executed and the number of viable oocytes collected, and rate of embryo production were determined. <h3>Results</h3> No clinical nor ultrasonographic alterations were detected in treated animals. Furthermore, similar numbers of viable oocytes, embryos produced, and rate of embryo production before and after MSC application were observed. <h3>Conclusion</h3> Based on these data, it can be concluded that the intra-ovarian application of 2.5 million adipose-derived MSC is safe. Future steps include the MSC treatment of individuals which present ovarian degeneration or lesions. <h3>Financial support</h3> This work was supported by CNPq, CAPES, FAPDF, FAPESP and Universidade of Brasilia.

Electronic Continuous Pain Measurement vs Verbal Rating Scale in gynaecology: A prospective cohort study

ObjectiveTo compare pain measured with a new electronic device – the Continuous Pain Score Meter (CPSM) – and the Verbal Rating Scale (VRS) during gynaecological procedures in an outpatient setting, and to correlate these outcomes with baseline anxiety and patient (in)tolerance to the procedure. Study designThis prospective cohort study was undertaken in two centres: a university hospital and a large teaching hospital in The Netherlands. Patients undergoing an outpatient hysteroscopy, colposcopy or ovum pick-up procedure for in-vitro fertilization in one of the two participating hospitals with availability of the CPSM were included. Pain was measured by both the CPSM and the VRS. Patient tolerance to the procedure was reported. Various outcomes of the CPSM were compared with those of the VRS and related to baseline anxiety scores. ResultsNinety-one of 108 included patients (84 %) used the CPSM correctly during the procedure, and it was possible to analyse the CPSM scores for 87 women (81 %). The CPSM scores were all linearly related to the VRS. The peak pain score on the CPSM (CPSM-PPS) had the strongest correlation with the VRS score for all three procedures. Higher CPSM-PPS was related to patient (in)tolerance to the procedure (p = 0.03–0.002). Anxiety at baseline was not correlated with pain perception, except for VRS during colposcopy (r = 0.39, p = 0.016). ConclusionThe majority of patients were able to use the CPSM correctly, resulting in detailed information on pain perception for each individual pain stimulus during three outpatient gynaecological procedures. The CPSM-PPS had the strongest correlation with the VRS score and patient (in)tolerance to the procedure.

228 Effect of seminal plasma on the interval to application of equine chorionic gonadotrophin for the recovery of cumulus-oocyte complexes in alpacas (Vicugna pacos)

Alpacas as other camelids are induced ovulators and require an external stimulus-mounting by the male-for ovulation. NGF-β, protein present in the seminal plasma (SP) is reported as being responsible for stimulating ovulation. However, limited information exists on the effect of ovulation inducers in the new follicular wave emergence with application on protocols of superstimulation. The aim of the study was to determine the effect of two ovulation induced techniques and two different times on the interval to application of equine chorionic gonadotrophin (eCG) for the ovarian superstimulation for the recovery of COCs by ovum pick- up (OPU). Alpacas were evaluated by transrectal ultrasonography with an Aloka SSD 500 ultrasound and transducer (7.5MHz) to determine the presence of a dominant follicle=7mm. A 2×2 experimental design was used with adult female alpacas, 6 to 8 years old, which were assigned to one of the treatments when a dominant follicle=7mm was present. Alpacas were assigned randomly 1 of 2 groups (Day 0) in which follicular ovulation was induced by seminal plasma 1mL IM (SP, n=13; group 1) with application of 650IU of eCG at 36h (n=6) or 48h (n=7) post-application of SP, or by GnRH with 0.008mg of Buserelin IM (GnRH, n=10; group 2) with application of 650IU eCG at 36h (n=5) or 48h (n=5) post-application of GnRH. On Day 7, COCs were counted and the OPU procedure was performed in every group. Data were analysed using ANOVA. Results are presented in Table 1. In conclusion, both inducers of ovulation and both times of application of eCG were effective for producing follicles of 7mm and COCs acceptable for recovery in Alpacas. Table 1.Number of follicles=7mm (top) and number of COCs recovered (mean±s.e.m.) Outcome and group 36 h 48 h Total Follicles GnRH 6.8±1.93 13.4±4.34 10.10±2.50a SP 5.5±2.23 5.29±1.15 5.38±1.25b Total 6.09±1.44 8.67±2.18 7.43±1.33 COCs GnRH 5±0.97 3±1.26 4.33±0.74a SP 1±0.41 0.33±0.22 0.67±0.25b Total 3.29±0.67x 1.4±0.6y 2.5±0.53 a,bValues within columns with different letters differ significantly (P=0.05). x,yValues within columns with different letters differ significantly (P=0.05). This research funded by CIENCIACTIVA-CONCYTEC as part of the project title “Role of seminal plasma in reproductive physiology and application of biotechnologies in camelids” (149-2017).

Superstimulation prior to the ovum pick-up improves the in vitro embryo production in nulliparous, primiparous and multiparous buffalo (Bubalus bubalis) donors

The aim of this study was to evaluate the ovarian follicular population, the oocyte yield and the in vitro embryo production (IVEP) of nulliparous (NU), primiparous (PR) and multiparous (MU) buffalo donors submitted to the superstimulation with FSH prior to the ovum pick-up (OPU). A total of 54 buffalo donors (18 NU, 15 PR and 21MU) received an intravaginal progesterone device (1.0 g) plus estradiol benzoate [2.0 mg, intramuscular (im)] at random stage of the estrous cycle (Day 0) during the breeding season (autumn and winter). Buffaloes from different categories were then randomly allocated to one of two groups (Control or FSH), in a cross-over experimental design. Buffalo donors in the Control group received no further treatment, whereas buffalo donors in the FSH group received a total dosage of 200 mg im of FSH on Days 4 and 5, in four decreasing doses 12 h apart (57, 57, 43 and 43 mg). On Day 7, the progesterone device was removed and the OPU procedure was performed in both groups. The same semen was used across all replicates and donor category. Data were analyzed by the GLIMMIX procedure of SAS 9.4®. There was no interaction between FSH treatment and animal category for all analyzed variables. Furthermore, no differences between animal category (P = 0.73) and FSH treatment (P = 0.53) were observed regarding the total follicles aspirated. However, the FSH treatment increased (P < 0.001) the proportion of large (>10 mm; FSH = 16.2% and Control = 2.0%) and medium-sized follicles (6–10 mm; FSH = 36.3% and Control = 6.1%) available for the OPU procedure. The total of recovered oocytes was greater in NU than in MU, and PR were similar to NU and MU (P = 0.05). No effect of FSH treatment was observed (P = 0.85) for this variable. Buffalo donors treated with FSH had a greater viable oocytes rate (P = 0.03), blastocyst rate (P = 0.03) and embryo yield per OPU-IVEP session (P = 0.07), however, no category effects were observed for these variables. These results provided evidence that superstimulation with FSH increased the proportion of large and medium-sized follicles available for the OPU procedure. Consequently, the FSH treatment enhanced the proportion of viable oocytes for culture and resulted in greater blastocyst rates and embryo yield per OPU-IVEP session in all buffalo donors categories.

115 Exosome-mediated microRNA expression profile in follicular fluid of metabolically divergent postpartum cows

Most high-milking cows enter a state of negative energy balance during the early lactation period. This phenomenon disturbs the metabolic status of the follicular fluid microenvironment, resulting in delayed ovulation. Cell-to-cell communication between the oocyte and the surrounding cells is crucial during folliculogenesis. Exosomes, evolutionarily conserved cargo molecules (30-150nm in diameter) carrying RNA and proteins, are known to be involved in cell-to-cell communication. Here, we aimed to investigate the association between postpartum metabolic status and the expression of exosomal microRNA (miRNA) in follicular fluid of Holstein-Friesian cows. For this, follicular fluid was collected from antral follicles (&amp;gt;8mm in diameter) using ovum pickup procedure from cows (n=30) on a weekly basis between weeks 5 and 10 postpartum. Follicular fluid collected from heifers (n=8) was used as a control. The energy status of each cow was assessed based on the blood metabolite (nonesterified fatty acids and β-hydroxybutyrate) concentration, body weight curve, and overall energy balance determined by dry matter intake. Afterwards, cows were categorized as early negative and late positive (cows show negative energy balance at early weeks and recovered at late weeks postpartum), always negative (cows did not recover until 15 weeks postpartum), and always positive (cows did not enter in to a state of negative energy balance). Following this, exosomes were isolated from pooled samples from each animal category using ultracentrifugation, and their morphology and size was characterised using electron microscopy and nanosight, respectively. Exosomal total RNA enriched with miRNA was isolated using an exosomal RNA isolation kit. Next-generation sequencing of miRNA was performed using Illumina NextSEqn 500 (Illumina Inc., San Diego, CA, USA). MicroRNAs with a fold change ≥2, P-value &amp;lt;0.05, and a false discovery rate of &amp;lt;0.1 were considered differentially expressed. The results showed that a total of 356 known and 156 novel miRNA were identified across samples. Differential expression analysis of miRNA between always-negative cows versus always-positive cows revealed down-regulation of all 6 differentially expressed miRNA, including bta-miR-451, bta-miR-132, and bta-miR-2285. Similarly, down-regulation of 14 miRNA, including bta-miR-20b, bta-miR-363, bta-miR-132, and bta-miR-451, and up-regulation of 3 miRNA was observed in always-negative cows compared to heifers. Furthermore, the target prediction analysis of the down-regulated miRNA have been shown to be involved in regulating different pathways including transforming growth factor-β signalling, cell cycle, hippo signalling, forkhead box O signalling, and endometrial cancer, among others. In conclusion, the results revealed that although negative energy balance in postpartum dairy cows suppressed exosomal miRNA expression in follicular fluid, the opposite was observed in metabolically unstressed cows. This divergence of exosome-mediated miRNA expression in the follicular fluid of metabolically stressed cows could be associated with the reduced fertility of those cows.

Changes in serum prolactin level during intracytoplasmic sperm injection, and effect on clinical pregnancy rate: a prospective observational study

BackgroundTransient hyperprolactinemia was proven to adversely affect the outcome of IVF. We aimed to identify changes in serum prolactin levels in patients undergoing ICSI, and to evaluate the effect of these changes on the clinical pregnancy rate.MethodsA prospective observational study included 90 patients scheduled for ICSI cycles. In each case 4 serum samples were collected during the cycle (midluteal, before ovum pick up procedure (OPU), 2 h after OPU, and before embryo transfer). Serum prolactin level was determined by immunoassay each time.ResultsThe sample collected 2 h after OPU had a mean difference of 25.8 ± 2.8 ng/ml compared to the basal serum prolactin (p < 0.01). In comparison to other samples, this highlighted a significant hyperprolactinemia occurring after OPU, and resolving before embryo transfer. No statistically significant difference between the different serum prolactin samples amongst the pregnant and non pregnant patients. There was a significant positive pearson correlation between the prolactin levels before OPU, and the presence of higher quality embryos (r = 0.274, p = 0.019).ConclusionIn normoprolactinemic women transient hyperprolactinemia is identified in patients undergoing ICSI, and it doesn’t affect the clinical pregnancy rates. A positive correlation was identified between higher quality embryos, and serum prolactin level before OPU.Trial registrationClinicalTrials.gov Identifier: NCT02292953, First received: November 10, 2014.

Improvement of bovine in vitro embryo production by ovarian follicular wave synchronization prior to ovum pick-up

This study evaluated the effects of the synchronization of ovarian follicular wave emergence on the efficiency of in vitro embryo production. Bos indicus cows (n = 20) were divided into two groups (control vs. synchronization) and subjected to repeated ovum pick-up (OPU) sessions (8 replicates each, with an interval of 21 days in a 2 × 2 crossover design) and subsequent in vitro embryo production. Cows in the control group (n = 10) were submitted to OPU procedures without any stimulation every 21 days. Animals in the synchronization group received a protocol-based progesterone implant, estradiol benzoate and prostaglandin on a random day of the estrus cycle (Day 0) and the OPU was performed on Day 5. After in vitro production, embryos were transferred to recipients synchronized at a fixed time and the diagnosis was performed 60 days later. An evaluation of the parameters for each OPU session revealed that donors that received the synchronization protocol pre-OPU showed a greater number of embryos (5.9 ± 0.5 vs. 4.5 ± 0.4; P = 0.037), higher rate of embryo production (45.8% vs. 38.5%; P = 0.001) and higher mean number of conceptions per group (2.2 ± 0.2 vs. 1.6 ± 0.2; P = 0.07) in relation to the group that did not receive hormonal treatment. We concluded that synchronization of the follicular wave prior to OPU showed positive effects on in vitro embryo production as well as on pregnancy rates.

Puncture and drainage of the subordinate follicles at timed artificial insemination prevents the risk of twin pregnancy in dairy cows.

The objective of this study was to determine whether fluid drainage from the subordinate follicles by ovum pickup procedures prevents the risk of twin pregnancy without reducing the fertility of the cow. Lactating dairy cows with at least two follicles over 12mm diameter located one on each ovary and selected from synchronized groups for fixed-time insemination were assigned to a Control (n=49) or Drainage (n=49) group. The largest follicle was considered as the dominant follicle. Draining of all subordinate follicles (≥10mm) was performed in the Drainage group. All drained follicles developed as a corpus luteum 7days later. In the Control cows, the presence of two corpora lutea (55.1%) was lower (p=.003) than those in the Drainage cows (87.8%), whereas the incidence of twin pregnancies was 50% and 0% for the Control and Drainage groups, respectively (p<.001). Draining did not affect the pregnancy rate. These results indicate that puncture and drainage of the subordinate follicles at insemination may eliminate the risk of twin pregnancies and reduce the risk of subsequent pregnancy loss by increasing the incidence of additional corpora lutea.

The status of assisted reproductive technology in Korea in 2012.

ObjectiveThis study was designed to report the status of assisted reproductive technology (ART) therapy in South Korea between January 1, 2012 and December 31, 2012.MethodsA localized online survey, originally developed by the International Committee Monitoring Assisted Reproductive Technologies, was first launched and provided to all available ART centers via email in 2015. Fresh embryo transfer (FET) cases were categorized as standard in vitro fertilization, intracytoplasmic sperm injection (ICSI), or half-ICSI. Thawed embryo transfer (TET) and other related procedures, including surgical sperm retrieval, were surveyed.ResultsData from 33,956 ovum pick-up procedures were provided by 75 clinics in 2012. Of the 33,088 cycles in which ovums were retrieved, a complete transfer was performed in 90.5% (29,932 cycles). In addition, 10,079 FET cycles were confirmed to have resulted in clinical pregnancy, representing a pregnancy rate of 30.5% per ovum pick-up and 33.7% per ET. The most common number of embryos transferred in FET was 2 (41.6%), followed by 3 (34.0%), and non-elective single ETs (10.0%). Of the 10,404 TET cycles in which transfer was completed, 3,760 clinical pregnancies (36.1%) were confirmed by ultrasonography.ConclusionThe overall clinical pregnancy rate for FET and TET cycles in 2012 was higher than in 2011 (33.7% vs. 33.2% and 36.1% vs. 31.1%, respectively). The most common number of embryos transferred in FET cycles was 2, unlike in 2011.

Beef donor cows with high number of retrieved COC produce more in vitro embryos compared with cows with low number of COC after repeated ovum pick-up sessions

The aim of the present study was to evaluate whether repeated ovum pick-up (OPU) procedures may affect the efficiency of OPU programs for in vitro embryo production (IVEP) in Bos indicus (Nelore) donors. In addition, the repeatability (r) efficiency of IVEP was also assessed. Data available were from 432 OPU-IVEP sessions that were performed at random stages of the estrous cycle in 36 cycling, nonlactating Nelore donors. Semen from three Nelore bulls was used for the IVF. Donors were submitted to 12 consecutive OPU procedures, with an interval of approximately 30 days between sessions. Data were analyzed as repeated measures using the GLIMMIX procedure of SAS 9.3. Cows yielding ≥15 COCs were defined as “high” COCs and cows with less than 15 COCs were defined as “low” COCs donors. The number of COCs retrieved decreased over time in donors classified with high COCs and remained fairly steady in cows with low COCs at the beginning of the program (Psession*COC retrieved groups = 0.02). Moreover, the number of COCs retrieved (P < 0.0001), and number of blastocysts produced per OPU (P = 0.001) was greater for the high COCs donors compared with the low COCs category, and these results were consistent across OPU sessions. Interestingly, there was no effect of COCs category on the rate of blastocyst development (P = 0.83). In addition, number of blastocysts produced were not affected by repeated OPU (P = 0.37) and interactions between time and COC category (P = 0.72). Similarly, blastocyst rate was not affected by repeated OPU (P = 0.21) and interactions between time and COC category (P = 0.58). Despite of COC category of the donor cows, repeatability was high for the number of COCs retrieved (r = 0.81), number of blastocysts produced per OPU (r = 0.79), and blastocyst rate (0.69). In conclusion, overall numbers of COCs decreased over time in donors classified as having high COCs. However, cumulative amounts of produced blastocysts were greater in donors with high COCs. More importantly, high repeatability was observed in terms of IVEP efficiency. Therefore, IVEP programs can be significantly improved with more aggressive selection toward donors with greater numbers of COCs.

Does intrauterine injection of low-molecular-weight heparin improve the clinical pregnancy rate in intracytoplasmic sperm injection?

ObjectiveHeparin can modulate proteins, and influence processes involved in implantation and trophoblastic development. This study aimed to assess the improvement of clinical pregnancy and implantation rates after local intrauterine injection of low-molecular-weight heparin (LMWH) in patients undergoing intracytoplasmic sperm injection (ICSI).MethodsA randomised case/control design was followed in women scheduled for ICSI. The study arm was injected with intrauterine LMWH during mock embryo transfer immediately following the ovum pickup procedure, while the control arm was given an intrauterine injection with a similar volume of tissue culture media. Side effects, the clinical pregnancy rate, and the implantation rate were recorded.ResultsThe pregnancy rate was acceptable (33.9%) in the LMWH arm with no significant reported side effects, confirming the safety of the intervention. No statistically significant differences were found in the clinical pregnancy and implantation rates between both groups (p=0.182 and p=0.096, respectively). The odds ratio of being pregnant after intrauterine injection with LMWH compared to the control group was 0.572 (95% confidence interval [CI], 0.27−1.22), while the risk ratio was 0.717 (95% CI, 0.46−1.13; p=0.146). No statistical significance was found between the two groups in other factors affecting implantation, such as day of transfer (p=0.726), number of embryos transferred (p=0.362), or embryo quality.ConclusionIntrauterine injection of LMWH is a safe intervention, but the dose used in this study failed to improve the outcome of ICSI. Based on its safety, further research involving modification of the dosage and/or the timing of administration could result in improved ICSI success rates.

Effects of a high-energy diet on oocyte quality and in vitro embryo production in Bos indicus and Bos taurus cows

The effects of different dietary energy levels [100 and 170% for maintenance (M) and high energy (1.7M), respectively] on metabolic, endocrine, and reproductive parameters were evaluated in nonlactating Bos indicus (Gir; n=14) and Bos taurus (Holstein; n=14) cows submitted to ultrasound-guided ovum pick-up followed by in vitro embryo production. The oocyte donor cows were housed in a tiestall system and fed twice daily (0800 and 1600 h). Twenty-one days before the beginning of the experiment, the animals were fed with a maintenance diet for adaptation followed by the experimental diets (M and 1.7M), and each cow underwent 9 ovum pick-up procedures 14 d apart. The recovered oocytes were cultured in vitro for 7 d. We measured glucose and insulin concentrations and performed glucose tolerance tests and the relative quantification of transcripts (PRDX1, HSP70.1, GLUT1, GLUT5, IGF1R, and IGF2R) from the oocytes recovered at the end of the experimental period. No interactions were observed between the effects of genetic groups and dietary energy level on the qualitative (viable oocytes, quality grade, and oocyte quality index) and quantitative (oocytes recovered) oocyte variables. There were no effects of dietary energy level on the qualitative and quantitative oocyte variables. However, Bos indicus cows had greater numbers of recovered structures, viable oocytes, and A and B oocyte grades as well as better oocyte quality index scores and lower DNA fragmentation rates compared with Bos taurus donors. In vitro embryo production (cleavage and blastocyst rates and number of embryos) was similar between diets, but the 1.7M diet reduced in vitro embryo production in Bos indicus cows after 60 d of treatment. Moreover, Bos indicus cows on the 1.7M diet showed lower transcript abundance for the HSP70.1, GLUT1, IGF1R, and IGF2R genes. All cows fed 1.7M diets had greater glucose and insulin concentrations and greater insulin resistance according to the glucose tolerance test. In conclusion, increasing dietary energy did not interfere with oocyte numbers and quality, but the 1.7M diet reduced in vitro embryo production in Bos indicus cows after 60 d of treatment. Finally, Bos indicus cows had greater oocyte quality, greater numbers of viable oocytes and greater in vitro embryo yield than Bos taurus.

Anti-Muellerian hormone levels in plasma of Holstein-Friesian heifers as a predictive parameter for ovum pick-up and embryo production outcomes.

The aim of this study was to investigate whether plasma anti-Muellerian hormone (AMH) levels of Holstein-Friesian heifers could be used to predict ovum pick-up (OPU) and embryo production outcomes. Plasma samples and data were collected from 64 heifers, which underwent repeated OPU with subsequent in vitro embryo production followed by embryo flushing after superovulation. AMH levels were significantly positively correlated with the number of follicles aspirated per OPU session (r = 0.45), recovered oocytes per OPU (r =0.43) and in vitro produced embryos per OPU (r = 0.28). No significant correlations between AMH and in vivo produced embryos were ascertained. Our results suggest that correlations between AMH and outcomes of an OPU-IVF program are too low to use AMH as a precise predictive parameter for the success of a particular OPU procedure in Holstein-Friesian heifers. However, AMH can help to identify groups of very good or very poor oocyte donors.