Total Number Of Embryos Research Articles

The progesterone prime protocol: an affordable option for ICSI in Egypt

BackgroundThe two main input indicators needed to assess the success of assisted reproductive technology (ART) strategies are cost-effectiveness and clinical outcome. Therefore, we focused in our study on the direct cost associated with the Progesterone Prime Ovulation Induction Protocol (P-P-OP protocol) compared to other protocols, and the clinical outcomes of this strategy were measured in their effect on implantation and pregnancy rates.MethodsA pilot single-armed study was conducted on 47 infertile women coming for an ICSI cycle. Progesterone priming was started with 10 mg dydrogesterone along with HMG 150–300 IU from the second day of the menstrual cycle and was maintained until the day of ovulation trigger. The primary outcome was the cost per cycle, while the secondary outcome was the clinical pregnancy rate. The cost per cycle, including the cryopreservation cost, was calculated for economic evaluation.ResultsPer each cycle, the number of retrieved oocytes was 14.87 ± 8.09, the number of mature oocytes was 9.81 ± 4.67, the number of total embryos was 8.74 ± 4.28 (grade A = 67.40%, grade B = 11.92%, grade C = 20.68%), and the number of transferred embryos was 2.43 ± 0.68. The clinical pregnancy rate in our study was 42.5%. Economically, the cost of the P-P-OP protocol for each patient was only 56 USD.ConclusionThe P-P-OP protocol is a simplified approach suitable for freeze-only strategies, offering the benefits of positive results, fewer injections, lower costs, and increased patient convenience.

Effect of Gonadotropin-Releasing Hormone Treatment on Superovulation Response and Embryo Recovery in Holstein Heifers: A Field Trial at Zagros Milk and Meat Company, Iran

Background: Superovulation is a crucial component of assisted reproductive technology. Inducing superovulation with Gonadotropin-releasing hormone (GnRH) can lead to ovarian hyperstimulation, potentially affecting reproductive outcomes. Objectives: This study aimed to examine the effects of GnRH treatment on superovulation response and embryo recovery in Holstein heifers. Methods: Twenty-one Holstein heifers (age: 135.15 months; weight: 361.05 kg) were selected based on their general health status and ovarian function. All heifers received two consecutive doses of prostaglandin F2α and underwent superovulation. The heifers were inseminated twice: Once at the onset of standing estrus and again 12 hours later. In the treatment group (13 heifers), a single dose of GnRH was administered simultaneously with the second insemination. The superovulation response was evaluated based on the number of corpus luteum (CL), unovulated follicles (UoF), total recovered embryos/ova, and the number of transferable embryos, using logistic regression with the GENMOD procedure in SAS. Results: The mean number of CL was not significantly greater in the control group (13.6) compared to the GnRH group (11.4). The average number of UoF was similar between the two groups (P = 0.1853). However, the control group had a significantly higher average total number of recovered embryos/ova (7.7) compared to the GnRH group (2.1). Additionally, the control group produced more transferable embryos, with an average of 2.5, while the GnRH group averaged 0.7 (P < 0.0062). Conclusions: Gonadotropin-releasing hormone likely deactivated the oviduct by disrupting the balance of estradiol and estrogen, leading to a reduction in both the total number of embryos and the number of transferable embryos in heifers.

Effect of the Application of PGF2α Associated With Ovulation Induction in a Fixed-Time Superovulation Programme for Precocious Nellore Heifers.

This study evaluated the effect of prostaglandin F2α (PGF2α) associated with gonadotropin-releasing hormone (GnRH) for ovulation induction in precocious indicus heifers submitted to a fixed-time superovulation (SOV) programme. Precocious Nellore heifers (n = 35), aged 13 months, were subjected to the SOV protocol. On day 0 (D0), all animals received intravaginal insertion of a progesterone (P4) device along with intramuscular administration of 2 mg of oestradiol benzoate, plus 200 IU of follicle-stimulating hormone in decreasing doses, with 12-h intervals between D4 and D7, in addition to 150 μg of D-cloprostenol on D6 and device removal on D7. On D8, the donors received 10.5 μg of buserelin acetate and the treatment group received 300 μg of D-cloprostenol/PGF2α. Artificial insemination was performed 12 h and 24 h after GnRH administration using frozen semen. On D15 of the protocol (i.e., D7 after insemination), the embryos were collected and evaluated. All animals passed through the control and treatment groups. Results were evaluated by analysis of variance using an adjusted mixed-effects model (p < 0.05). There was no difference in the total number of embryos between the control and treatment groups (10.40 ± 1.52 vs. 9.60 ± 1.36; p = 0.63) or viable embryos (6.30 ± 1.22 vs. 4.30 ± 0.71). For precocious indicus heifers, treatment with PGF2α in association with GnRH did not affect embryo production in the fixed-time SOV protocol.

Vaginal Progesterone Gel versus Intramuscular Progesterone for Luteal Phase Support in Suboptimal Responders Undergoing Assisted Reproductive Cycles

Background: The purpose of this research is to compare the efficacy of 8% micronized vaginal progesterone gel (VAG) at 180 mg daily versus intramuscular progesterone (IMP) in oil at 100 mg daily for luteal phase support (LPS) after fresh single embryo transfer (SET) in Patient-Oriented Strategies Encompassing Individualized Oocyte Number (POSEIDON) Group 1b patients, and to ascertain whether the type of LPS predicts live birth in these patients. Methods: A total of 2420 assisted reproductive technology (ART) cycles performed in Ondokuz Mayıs University IVF Unit were analysed retrospectively. The data of POSEIDON Group 1b (unexpected suboptimal responders) who underwent antagonist in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) cycles were included. All patients were categorized into groups according to the form of LPS, specifically VAG and IMP. Pregnancy, clinical pregnancy rate (CPR), live birth rate (LBR), spontaneous abortion rate and predictors of live birth (age, infertility duration, body mass index (BMI), type of progesterone used for luteal support, serum day 3 estradiol, serum progesterone on the day of human coryonic gonadotrophin (HCG), total duration of induction, serum estradiol on the day of HCG, the total number of oocytes retrieved, number of mature oocytes obtained, average gonadotropin dose per day, total gonadotropin dose administered and total number of embryos retrieved) were analyzed. Results: There was no statistically significant difference between the micronized VAG and IMP groups in terms of age (31 (23–35) vs. 27 (23–35), p = 0.319). There were no statistically significant differences in pregnancy outcomes between the two groups concerning CPR per transfer (70.6% vs. 75.6%; p = 0.364), ongoing pregnancy per cycle (36.2% vs. 39.5%; p = 0.577), and LBR per cycle (34.4% vs. 36.1%; p = 0.785). The spontaneous abortion rates (36.2% vs. 36.8%; p = 0.921) were similar between VAG and IMP groups. The type of LPS did not emerge as a parameter predicting pregnancy (odds ratio (OR): 0.718, 95% confidence interval (95% CI): 0.652–1.313, p = 0.451), clinical pregnancy (OR: 0.598, 95% CI: 0.592–1.289, p = 0.562) and live birth (OR: 0.802, 95% CI: 0.661–1.202, p = 0.580). The logistic regression analysis aimed at assessing the influence of confounding factors, namely age, BMI, and duration of infertility on pregnancy rate, CPR and LBR, did not reveal statistically significant results (p &gt; 0.05). Conclusions: VAG 180 mg daily provide similar pregnancy outcomes compared to 100 mg daily IMP in POSEIDON Group 1b patients undergoing antagonist fresh IVF/ICSI cycles.

Constructing a predictive model for live birth following fresh embryo transfer in antagonist protocol for polycystic ovary syndrome.

The present research aims to assess the factors that influence live birth outcomes following fresh embryo transfers using antagonist protocols in individuals diagnosed with polycystic ovary syndrome (PCOS). Furthermore, it seeks to develop a predictive nomogram model to facilitate clinical decision-making and provide personalized treatment strategies. This retrospective cohort research analyzed the clinical data of 1242 individuals having PCOS who went through fresh embryo transfers employing antagonist protocols and in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) at Fujian Provincial Maternal and Child Health Hospital between January 2018 and December 2022. Individuals were assigned randomly to a modeling group (869 cases) and a validation group (373 cases) in a 7:3 ratio. The Boruta algorithm and multivariable logistic regression were utilized to identify independent risk factors linked to live births after transfer. A predictive nomogram was subsequently developed. The discriminatory power of the model and its accuracy were monitored by utilizing receiver operating characteristic (ROC) curves, calibration curves, and decision curve analysis. Multivariable logistic regression analysis identified several independent factors that influence live birth rates in fresh embryo transfer cycles for individuals having PCOS using antagonist protocols, including female age, body mass index (BMI), infertility duration, serum testosterone levels, progesterone levels at the time of human chorionic gonadotropin (hCG) injection, number of high-quality cleavage-stage embryos, type of embryo transferred, and the total number of embryos transferred. Based on these findings, a predictive nomogram was developed. The area under the ROC curve stood at 0.804 (95% confidence interval (CI), 0.775-0.833) for the modeling group and 0.807 (95% CI, 0.762-0.851) for the validation group. Calibration curves confirmed that the predictions of the nomogram closely matched the actual live birth outcomes. Decision curve analysis highlighted that the model provides significant net benefits for predicting live birth rates, with optimal performance across a probability range of 16.5 to 88.6%. Independent factors, including female age, infertility duration, BMI, serum testosterone levels, progesterone levels on the day of hCG injection, and the number and type of high-quality cleavage-stage embryos transferred are pivotal in influencing live birth outcomes in fresh embryo transfer cycles under antagonist protocols in individuals with PCOS undergoing IVF/ICSI treatments. The predictive nomogram developed from these factors offers substantial predictive accuracy and clinical utility, providing a reliable basis for clinical prognosis, targeted interventions, and the development of personalized treatment plans.

Influence of male body mass index on semen analysis parameters and in vitro fertilization outcomes.

Concurrent global increase of prevalence of obesity and male fertility implies link between overweight and obesity with male subfertility. This hypothesis is supported by numerous population-based epidemiological studies. Increase in body mass index (BMI) is associated with poor sperm quality in fertile, and more noticeable in infertile men. Nevertheless, some studies disprove damaging effect of BMI on semen quality. To examine the influence of men's BMI in infertile couples undergoing in vitro fertilization (IVF) on semen analysis parameters and IVF outcomes. Study encompassed all couples who underwent IVF at Gynecology and Obstetrics Clinic Narodni Front in Belgrade during 2018 and 2019. Exclusion criteria were azoospermia, conditions and diseases that could affect the semen analysis parameters (diabetes, malignant diseases treated with radiation and/or chemotherapy, trauma or surgery of the genital organs, mumps or undescended testicles in childhood). Evaluated semen analysis parameters included semen ejaculate volume, sperm pH, sperm count, sperm motility, and sperm morphology. IVF outcomes comprised total number of embryos, number and percentage of obtained good-quality embryos and clinical pregnancy rates. Based on BMI value, participants were divided into a group of underweight (Group 1), normally weight (Group 2), overweight (Group 3), and obese men (Group 4). After applying inclusion and exclusion criteria, 411 men (couples) were included in the analysis. The largest number of men were overweight, while the smallest belonged to the group of underweight participants. There are no significant differences in the semen analysis parameters between study groups. Correlation analysis shown weak and insignificant correlation between BMI and semen analysis parameters. The number and proportion of good quality embryos is significantly lower in overweight and obese study groups compared to normal weight and underweight groups (2.89, 2.91, 2.42, and 2.36, respectively, P = .041). The differences in other IVF outcomes: total number of embryos (3.61, 3.74, 3.21, and 3.37, respectively) and clinical pregnancy rates (41.26%, 43.09%, 42.78%, and 39.95%, respectively) between study groups were not significant (P > .05). BMI does not significantly affect semen analysis parameters, but a higher BMI is associated with a lower number and proportion of good quality embryos in IVF outcomes.

P-669 Not the oocyte yield but the number of embryos predicts live birth at high response: analysis of 39,136 fresh IVF/ ICSI cycles from National Database

Abstract Study question Oocyte or embryo yield- which is a better predictor of live birth (LB) in fresh cycle IVF/ ICSI? Summary answer The embryo yield is better than oocyte in predicting fresh-cycle LB and positively correlates with LB at high ovarian response when the oocytes yield doesn’t. What is known already As the LB rates plateau after a certain oocyte yield, the number of oocytes collected no longer can predict the live birth in IVF/ ICSI cycles with fresh embryo transfer.Since pregnancy outcomes also depend on the number of mature oocytes obtained, fertilisation rate and subsequent embryo development, it can be postulated that the number of total or transferrable embryos could be a better predictor of LB. Study design, size, duration Retrospective analysis of 39,136 fresh IVF/ ICSI cycles that met the inclusion criteria from the national database published by Human Fertilisation and Embryology Authority (HFEA) in the United Kingdom. We analysed 2 years of published data between January 2015 and December 2016. Participants/materials, setting, methods Only the first cycles of IVF/ ISCI that had oocyte retrieval procedure with intended fresh embryo transfer were included. Couples with any cause of infertility and female partners aged up to 44 years were included. Very few cycles with &amp;gt;25 oocytes had fresh transfer, therefore excluded. The ovarian response has been graded by 0-5, 6-10, 11-15, 16-20, 21-25 collected oocytes. Significant confounders found in univariate analysis were adjusted for in the multiple regression model. Main results and the role of chance Women’s age, method of insemination (IVF or ICSI) and number of transferred embryos and stage of transfer significantly associated with LB in univariate analysis (p &amp;lt; 0.0001 each). The oocyte yield predicted fresh-cycle LB only in low response group (0-5 oocytes), but not in higher response groups. Total number of embryos, the number of transferable embryos and embryo utilisation rate remained positively correlated with LB in all response groups. The number of transferable embryos found to be the best predictor of LB with the highest adjusted OR 1.51 (CI:1.4-1.65, p &amp;lt; 0.0001) in the lowest response (0-5 oocyte) group, that gradually declined to 1.06 (CI:1.03-1.10, p = 0.0003) in 21-25 oocytes group. Followed next by the total number of embryos with the highest adjusted OR of 1.20 (CI:1.13-1.27, p &amp;lt; 0.0001) in the lowest response (0-5 oocytes) group, down to 1.04 (CI:1.02-1.06, p = 0.0003) in the 21-25 oocytes group. The number of oocytes was the weakest in predicting LB (adjusted OR 1.07 (CI 1.02-1.12; p = 0.003) in the 0-5 oocytes group only. The inference remained the same when ICSI cycles with supposedly sperm factor was excluded. Limitations, reasons for caution Information on patient’s BMI or stimulation dose is not available in the HFEA database. However, it might have little impact in our findings, given large sample size. Ages are given in groups, so the groups taken as ordinal (dummy) variables in multiple regression analysis. Cumulative LB data were not available. Wider implications of the findings Having been influenced by both oocyte and sperm competency, the number of transferable embryos may have a stronger relation with the pregnancy outcomes than merely the oocyte number. On theory, this may be applicable for cumulative LB too, but awaiting confirmation by data evidence. Trial registration number Not applicable

P-248 Egg vitrification related to female age: survival and laboratory outcome of 1233 vitrified-thawed oocytes in non-medical autologous use

Abstract Study question Is the female age at retrieval a determinant factor for the viability and potential of the thawed eggs? Summary answer In patients over 40, thawed oocytes display more abnormal fertilization, develop less in extended culture and yield less top quality blastocyst. What is known already Oocyte cryopreservation is a well-established procedure especially since the advent of the vitrification technique. The successful outcomes achieved with vitrification encourage its application in fertility preservation (FP) for medical and non-medical reasons. Besides, the age-related decline in women’s fertility and the oocyte quality negatively affect the outcome of assisted reproduction. Some studies have reported that age at oocyte retrieval strongly influences reproductive prognosis after FP. However, the appropriate age for non-medical FP is still debated. Although oocyte freezing at a younger age is recommended, there should be a balance between a favourable outcome and the cost-effectiveness of the strategy. Study design, size, duration Retrospective study on 166 patients with frozen-thawed oocytes who underwent ICSI during 2021-2023 in a single hospital. Oocyte vitrification had different purposes: banking for poor responders, to limit the number of embryos, and to solve unexpected situations where partner’s semen sample was unavailable. Patients were divided into three groups according to their age at retrieval (Group A ≤ 37 n = 77, Group B 37-40 n = 44, Group C ≥ 40 n = 45). Post-warming survival, fertilization, and embryological data were studied. Participants/materials, setting, methods 1233 oocytes were thawed. The intact oocytes were microinjected 1-2 hours after warming. Embryos were cultured in Sage 1-StepTM to blastocyst stage, and rated on daily basis. Embryo transfer was performed on day 3 or day 5 and remaining blastocysts were frozen on Day 5 or Day 6. Post warming survival, lysis after ICSI, fertilization, embryo development, blastulation rate (BR) and top blastulation rate (TBR), and morphokinetic features were studied in the 3 groups. Main results and the role of chance 84.6% of the 1233 oocytes (vitrified in a high-security closed system) survived the warming process. 1043 eggs were injected and the post-ICSI lysis rate was 11.2%. No significant differences were seen in thawing survival rate in groups A, B and C (85.6% vs. 83.5% vs. 83.8%, p=0.635) nor in the post-ICSI lysis rate (13.1% vs. 7.4% vs. 11.2%, p=0.072). 2PN fertilization rate was similar between the groups (67.8% vs. 61.3% vs. 67.7%, p=0.178) as well as the total number of embryos (303 vs 146 vs 188 p=0.605). However, triploid fertilization rate (3PN) statistically grew with age (2.8% vs. 5.8% vs. 6.5%, p=0.034). Significant differences were also reported in the global abnormal fertilisation rate (1PN+3PN) (6.4% vs 10.3% vs. 13.3%, p= 0.005). BR and TBR were statistically lower after the age of 40 (BR: 61.5% vs. 62.7% vs 40% p&amp;lt;0.001, TBR: 43.3% vs. 47.9% vs. 24.3% p&amp;lt;0.001). Morphokinetic features in the 3 groups were studied (abnormal cleavage pattern, tSB and tB). Overall results were similar between group A and group B but oocyte potential decreases drastically from the age of 40. Limitations, reasons for caution The major limitation of the study is its retrospective nature and the relative small size of each group. As most of patients were transferred during 2023, the pregnancies have not delivered yet so the live birth rate is not mentioned. Wider implications of the findings Studies have reported the impact of age at oocyte retrieval in case of medical FP in cancer and endometriosis. Here we report a negative effect on outcome after the age of 40 in non-medical FP. Patients should be informed that FP is not a certification of a successful outcome. Trial registration number not applicable

O-009 Comparison of the flexible Progestin-Primed Ovarian Stimulation with the GnRH antagonist protocol in predicted suboptimal responders: a freeze-all randomized controlled trial

Abstract Study question Is the Flexible Progestin-Primed Ovarian Stimulation (fPPOS) non-inferior to GnRH antagonist regarding fertility outcomes in women with a suboptimal response scheduled for freeze-all cycles? Summary answer In predicted suboptimal responders, who do not contemplate a fresh embryo transfer, the ongoing pregnancy outcomes of fPPOS was comparable to that of GnRH-antagonist protocol. What is known already Recent retrospective cohort studies involving donor cycles have demonstrated that Medroxyprogesterone Acetate (MPA) can be effectively and flexibly employed during the mid-follicular phase of the PPOS protocol. This approach prevents premature ovulation without compromising oocyte yields. Given its lesser degree of pituitary suppression, the fPPOS protocol may be particularly relevant for women predicted to have a suboptimal response (defined as AFC &amp;lt; 10) where accumulation of more oocytes could enhance cumulative success rates. Study design, size, duration This is a open-label RCT, performed in a single IVF centre Between July 2019 and June 2023. A total of 484 women scheduled for freeze-all strategy were randomly assigned to fPPOS or GnRH-antagonist protocol. The primary outcome was live birth per woman starting stimulation following18 months of randomization. In this abstract, we present fertility outcomes following the initial frozen embryo transfer (FET) and ongoing pregnancy rates per women within a period of 6 months. Participants/materials, setting, methods Women (AFC&amp;lt;10) referred for IVF/ICSI and did not intend to undergo a fresh transfer, were recruited for this study. MPA (10 mg/day) and Cetrotide (0.25 mg/day) were initiated either when the leading follicle reached 14 mm or on the sixth day of stimulation, whichever occurred first. Due to the COVID-19 pandemic, 22 women withdrew from the study at the stimulation commencement, resulting in 462 women included in the analysis. Data underwent intention-to-treat and per-protocol analyses. Main results and the role of chance No significant difference was observed between the two groups in terms of serum endocrine profiles and stimulation parameters, including the consumption of gonadotropins. Within the 6-month period, 79.5% of women in the fPPOS group and 84.02% in the GnRH antagonist group had undergone at least one cycle of FET (P = 0.234).The ongoing pregnancy rates per women were 48.20% (107/222) in the fPPOS group versus 52.92% (127/240) in the GnRH-anta group (RR: 0.91, 95%CI, 0.76 to 1.09, P = 0.311). There are 62 women who are ongoing pregnancy but do not reached live birth during the follow-up period. Regarding the outcomes of the initial FET cycle, there was no difference in the ongoing pregnancy rate between the two groups (49.01% vs 48.37%, RR: 1.03, 95% CI, 0.81 to 1.26, P = 0.908). Other outcomes including biochemical pregnancy rate (62.25% vs 63.59%), clinical pregnancy rate (60.26% vs 57.61%), and miscarriage rate per pregnancy (16.48% vs 15.09%), also exhibited no significant differences. The number of oocytes retrieved, two-pronuclear oocytes, viable and high-quality embryos, as well as the total number of embryos cryopreserved were also similar between groups. There was one case of premature ovulation in the fPPOS group and none in the GnRH-ant group. Limitations, reasons for caution Due to the distinct administration routes of the medications, blinding was not feasible in this open-label trial, which may affect the assessment of outcomes. All participants were recruited in a single center from one country, limiting the generalizability. In this abstract, we only report on the ongoing pregnancy outcomes. Wider implications of the findings This is the first freeze-all RCT comparing the long-term outcomes of fPPOS and GnRH antagonist in autologous ovarian stimulation cycles. Eliminating the variability of fresh and frozen transfers, the fPPOS seems an efficacious alternative for women with predicted suboptimal response, when a GnRH antagonist would have been the preferred regimen. Trial registration number ChiCTR2000030356

P-595 Increased clinical pregnancy rates using a clinical decision support tool - a retrospective cohort study

Abstract Study question Does using a clinical decision support tool, Opt-IVF, to guide hormone dosing during superovulation yield increased clinical pregnancy rates in routine clinical practice? Summary answer Higher clinical pregnancy rates were obtained using Opt-IVF to guide stimulatory hormone dosing during superovulation. What is known already ART success rates and cost depend heavily on the superovulation stage of the cycle. Two prospective single-center trials and one prospective multi-center trial had shown that using Opt-IVF to guide dosing resulted in increased numbers of high-quality embryos, used lower hormone doses, and required less ultrasound testing. This is a retrospective cohort study of the use of Opt-IVF in clinical practice over 12 months compared with the control cohort from the previous 12 months. Study design, size, duration Design: The study examined patients undergoing an IVF cycle using Opt-IVF in a single center for 12 months. Opt-IVF was used in all patients who presented to one of the investigators at Akanksha Hospital during the study period. As a comparison group, we examined patients from the previous 12 months when Opt-IVF was not used, and standard clinical protocols were followed to guide dosing. Duration: The study was conducted over 24 months Participants/materials, setting, methods 204 women aged 25-45 years underwent superovulation using hormone doses suggested by Opt-IVF (intervention). Of the participants, 19% were poor responders, 74% were below 35 years of age, 19% were 35-39 years of age, and 7% were above 39 years of age. The control group included 207 women, of whom 10% were poor responders. 72% were below 35 years of age, 26% were 35-39 years, and 2% were above 39 years of age. Main results and the role of chance The participants in the intervention group required significantly lower cumulative gonadotropin dosage compared to the control group during their controlled ovarian stimulation cycles (p &amp;lt; 0.0001). The average number of total embryos was 20% higher (p &amp;lt; 0.001), and the average number of good-quality embryos was 50% higher (p &amp;lt; 0.0001) in the Opt-IVF group than in the control group. Only 4.9% of cycles were canceled in the intervention group, compared to 12.9% canceled in the control group. Overall, Clinical pregnancy rates (including canceled cycles) increased from 35 % in the control group to 43% in the intervention group. The clinical pregnancy rate for poor responder patients was 39% in the Opt-IVF group but was only 16% in controls (p &amp;lt; 0.0001). Similarly, the clinical pregnancy rates were 34% in patients ages 35-39 years and 33% in patients above 39 years of age, much higher than the corresponding clinical pregnancy rates in controls (27% and 0%, respectively). Higher clinical pregnancy rates were also noted in the intervention group for normal responders and for younger patients. Limitations, reasons for caution This is a single-center retrospective cohort study. Wider implications of the findings Using Opt-IVF to guide dosing resulted in increased numbers of high-quality embryos and higher clinical pregnancy rates, particularly among older patients and poor responders. The proportions of these patients can be expected to grow in the future, so improving results in such patients is essential. Trial registration number not applicable

O-186 A novel micro-insemination method, assisted sperm fusion insemination, increases the embryo utilization rate compared to conventional intracytoplasmic sperm injection

Abstract Study question Does a novel micro-insemination method, assisted sperm fusion insemination (ASFI), improve fertilization and embryonic development compared to conventional intracytoplasmic sperm injection (C-ICSI)? Summary answer ASFI showed comparable fertilization and blastocyst formation rates to C-ICSI, while achieving significantly higher embryo utilization rates. Furthermore, there were no degenerated oocytes after ASFI. What is known already We have reported that ASFI yielded fertilization and blastocyst rates similar to those obtained with rescue ICSI performed at 6 hours after in vitro fertilization (IVF). In ASFI, fertilization is achieved by pressing a zona pellucida (ZP)-bound sperm onto the oocyte membrane without breaking the oocyte membrane. However, there was a risk of artificial polyspermy because the sperm were pressed against the unfertilized oocytes after IVF. Moreover, it is uncertain whether fertilization occurred with sperm that penetrated into the oocyte via IVF or that were pressed against the oocyte membrane. Clarifying this requires performing ASFI of the oocyte without IVF. Study design, size, duration We prospectively evaluated the effectiveness of ASFI in a sibling oocyte study including 14 subjects from which at least two metaphase II (MII) oocytes and one immature or degenerated oocyte were retrieved. Between December 2022 and October 2023, 119 MII oocytes were obtained in 22 oocyte retrieval cycles and used in the study. Sibling MII oocytes were randomly assigned to a C-ICSI or ASFI group. The present study was approved by the local ethics committee. Participants/materials, setting, methods In the ASFI group, 10,000 motile sperm were incubated with at least one immature or degenerated oocyte for 3 hours in 100 μL of medium per patient. After incubation, the motile sperm bound to the ZP were aspirated directly using an injection pipette. The heads of these ZP-bound sperm were pressed onto the membrane of MII oocytes for 10 seconds using the injection pipette. C-ICSI was conducted using the conventional method. Main results and the role of chance The mean and standard deviation of age of the women and number of MII oocytes per cycle were 39.9 ± 2.2 years and 5.5 ± 1.8, respectively. All sperm collected from the ZP adhered to the MII oocyte membrane after ASFI. Furthermore, time-lapse imaging revealed that all sperm adhering to the MII oocyte membrane were subsequently incorporated into the oocyte. Comparison of the ASFI and C-ICSI groups did not reveal any significant differences in 2 pronuclear embryo rate (ASFI group: 85.4% vs. C-ICSI group: 71.8%), degeneration rate (0% vs. 5.6%), good-quality embryo rate at day 3 (63.4% vs. 52.9%), blastocyst formation rate (67.6% vs. 51.9%), or good-quality blastocyst (Grade 3BB and above by the Gardner criteria) formation rate (29.4% vs. 15.4%). On the contrary, the embryo utilization rate (defined as the total number of embryos transferred and cryopreserved divided by the number of MII oocytes) was significantly higher in the ASFI group compared to the C-ICSI group (47.9% vs. 25.4%, P &amp;lt; 0.05). Limitations, reasons for caution This study was limited by using the ZP for sperm selection. To expand its applicability, a method to identify sperm capable of fusing with an oocyte without using the ZP must be established. Additionally, since our sample size was small, larger studies are required to confirm our findings. Wider implications of the findings ASFI is expected to enhance the survival rate of oocytes and to increase the number of embryos available for implantation, not only compared to rescue ICSI but also C-ICSI. ASFI could aid in understanding fertilization mechanisms by enabling direct observation of oocyte-sperm fusion during the process. Trial registration number UMIN000050184

P-428 Identifying key factors for achieving biochemical pregnancy in first autologous IVF cycles: an artificial intelligence-assisted approach

Abstract Study question What key factors determine a positive biochemical pregnancy outcome in first autologous IVF cycles, as identified by machine learning (ML) algorithms? Summary answer Maternal age, AMH, BMI, number of day 3 embryos and total embryos transferred emerged as top predictors for biochemical pregnancy across several ML methods. What is known already ML methods are powerful tools that discern patterns from multiple features to predict outcomes. Beyond their clear predictive capabilities, ML algorithms also offer valuable clinical insights by uncovering underlying data trends. These insights come from understanding the algorithms’ decision-making processes, revealing the rationale behind their predictions. While maternal age is known to influence IVF success, other pre-treatment and in-cycle factors also play a significant role. Understanding these factors can enhance patient management and guide clinical advice and interventions. Study design, size, duration This multicenter retrospective study included data from 9,945 cycles performed between January 2010 and July 2021. Only first autologous cycles using fresh oocytes and donor or patient sperm (fresh or frozen) were included. Preimplantation genetic testing (PGT) cycles were excluded. Both pre-treatment and in-cycle variables were considered, with treatment outcome expressed as cumulative biochemical pregnancy. Cases and variables deemed to have poor data quality were excluded. The final dataset comprised 80 variables and 7350 cycles. Participants/materials, setting, methods Five ML classifier algorithms were applied to predict cumulative biochemical pregnancy using pre-treatment and in-cycle variables. The models tested included logistic regression, support vector machine, random forests, gradient boosting and multilayer perceptron. Shapley values analysis was performed to explain the decision-making structures of all the algorithms tested, quantifying the impact of each included feature on outcome prediction. Main results and the role of chance Within our cohort, mean maternal age was 37.3±4.7 years, mean serum AMH was 2.4±2.1 ng/ml, mean BMI was 23.7±4.0 and mean antral follicle count (AFC) was 11.6± 7.6. Notably, 40.1% of cycles used donor sperm, while 33.8% of male patients were normozoospermic. The majority of cycles (90.8%) used antagonist stimulation protocols, yielding an average of 7.8±5.5 oocytes per patient. Fertilization rate was 70.6%, with a mean of 2.7±1.8 usable embryos obtained per cycle. Fresh embryo transfers accounted for 73% of cases, with an average of 1.5±0.9 transfers performed per cycle. The embryo utilization rate was 64.6% and 48.1% of cycles resulted in a biochemical pregnancy. Among the evaluated algorithms, the gradient boosting model demonstrated superior performance, with an accuracy of 70.6±1.4% and AUC of 77.3±1.7%. Shapley values across all five algorithms consistently revealed the negative correlation between maternal age and biochemical pregnancy rates. Among pre-treatment variables, AMH levels had a positive influence on outcomes, followed by BMI, AFC, and parity status. Regarding in-cycle variables, the number of embryos available at day 3 and the total number of embryos transferred, positively affected the chances of pregnancy. The number of mature oocytes was also an important in-cycle predictive feature. Limitations, reasons for caution his retrospective study spans an extended timeframe, suggesting potential variations in clinical and laboratory practices over the years. Therefore, caution is advised in generalizing the results to other patient populations or to contemporary data gathered using newer and enhanced clinical protocols. Wider implications of the findings The consistent identification of maternal age, ovarian reserve markers, and embryo quantity as primary predictors of pregnancy by the five AI models corroborates existing clinical understanding. This consistency emphasizes the strength and dependability of AI to reinforce and complement clinical knowledge with data-driven evidence. Trial registration number not applicable

The significance of light chronodestruction in the development of the embryotoxic effect in the experiment

Objective. To assess the significance of light chronodestruction in the development of the embryotoxic effect in the experiment. Materials and methods. The experiment was conducted on 60 female rats of the Wistar line. To assess the significance of chronodestruction in the development of embryotoxic effect, the uterus was isolated in the experiment, in which the number of implantation sites, the total number of embryos, the number of live and dead embryos, and the number of resorptions were estimated. The number of corpus luteum in the harvested ovaries was estimated. In addition, the mean embryo weight (g) and mean craniocaudal size (mm) were recorded. Indicators of embryotoxic effects of light deprivation were determined by pre- and post-implantation death, which was calculated in accordance with methodological recommendations. Results. Exposure to light deprivation reduces the number of implantation sites as of 7th, 14th and 21st days by 1,5-1,8 times, the total number of embryos by 1,6-1,8 times, the number of living embryos by 2,2-9 times and increases the number of dead embryos as of 14th and 21st days by 4-5,5 times, the number of resorptions – 1,6-11 times. The average craniocaudal size (mm) of embryos in experimental animals was recorded below control values by 1,5 times, 1,3 and 3,7 times by 7th, 14th, 21st days respectively. In females exposed to light deprivation, there was a significant increase in pre-implantation mortality by 35-41,8 times and post-implantation mortality by 7,2-20,4 times compared to the control. Conclusion. Light deprivation may have a negative effect on pregnancy and fetal development in female rats, which is confirmed by an increase in pre- and post-implantation mortality.

Comparison of pregnancy outcomes in women with normal ovarian response to the gonadotropin-releasing hormone agonist protocol using different trigger methods: a single-center retrospective cohort study based on propensity score matching.

To investigate whether gonadotropin-releasing hormone agonist (GnRH-a) combined with human chorionic gonadotropin (HCG) can improve pregnancy outcomes in patients with normal ovarian response (NOR). In this retrospective cohort study, data of 404 NOR patients undergoing fresh embryo transfer (ET) from 2018 to 2022 were studied. Patients were divided into HCG group and HCG plus GnRH-a group according to trigger methods. After confounding factors were controlled by propensity score matching, 67 cases were included in HCG group and HCG plus GnRH-a group, respectively, and pregnancy outcomes were assessed. Basal data, ovarian stimulation, embryological data and pregnancy outcomes were compared. The effect of trigger methods on pregnancy outcomes was analyzed by binary logistic regression. There was no statistically significant differences in embryological data, embryo implantation rate, clinical pregnancy rate, live birth rate of ET, number of fresh embryos transferred and total number of embryos transferred after one cycle of oocyte retrieval. While, cumulative live birth rate (CLBR) was better in the dual-trigger group than in the HCG group. The binary logistic regression analysis indicated that the trigger methods had an independent influence on embryo implantation and cumulative live birth. During IVF/ICSI, dual-trigger could potentially play a role in improving oocyte quality, ensuring embryo implantation rate, clinical pregnancy rate, live birth rate of ET and cumulative live birth rate at the end of one ovum pick-up (OPU) cycle, and reducing the physical, temporal and financial negative consequences due to repeated OPU cycle.

The impact of microfluidics sperm processing on blastocyst euploidy rates compared with density gradient centrifugation: a sibling oocyte double-blinded prospective randomized clinical trial

ObjectiveTo compare euploidy rates among blastocysts created from sibling oocytes injected with sperm processed by microfluidics or by density gradient centrifugation. DesignSibling oocyte randomized controlled trial. SettingSingle university-affiliated infertility practice. Patient(s)106 patients aged 18-42 undergoing fresh in vitro fertilization (IVF) cycles with preimplantation genetic testing (PGT) between January 2021 to April 2022 contributed 1442 mature oocytes which were injected with sperm processed via microfluidics or density gradient centrifugation. Intervention(s)Sperm sample divided and processed via a microfluidics device and density gradient centrifugation for injection into sibling oocytes. Main outcome measure(s)Primary outcome was the embryo euploidy rate. Secondary outcomes included the fertilization, high quality blastulation, and ongoing pregnancy rates. Result(s)The blastocyst euploidy rate per mature oocyte was not significantly different in the study group compared to the control group (22.9% versus 20.5%, p = 0.60). The blastocyst euploidy rate per biopsied embryo was also similar between the two groups (53.0% versus 45.7%, p = 0.34). However, the fertilization rate per mature oocyte injected was found to be significantly higher in the study group compared to the control group (76.0% versus 69.9%, p = 0.03). The high quality blastulation rate per mature oocyte injected was similar between the two groups, as was the total number of embryos frozen. There were no differences in the number of participants with no blastocysts for biopsy or the number of participants with no euploid embryos between the two groups. Among the male factor infertility and recurrent pregnancy loss subgroups, there were no differences in euploidy rates, fertilization rates, blastulation rates, or total numbers of blastocysts frozen, although the study was underpowered to detect these differences. Seventy-seven patients underwent frozen embryo transfer; there were no significant differences in pregnancy outcomes between the two groups. Conclusion(s)Microfluidics processing did not improve embryo euploidy rates compared to density gradient centrifugation in this sibling oocyte study, although fertilization rates were significantly higher.

The Reproductive Biology of Puye (Galaxias maculatus) under Experimental Culture.

This study determines the reproductive patterns of puye (Galaxias maculatus) under culture conditions. A population of 567 wild fish was caught in the Cautín River, Chile, and held in captivity for four years. Mortality, sex ratio, gonadosomatic index (GSI), sexual maturity stages, spawning period, type and frequency of spawning, and fecundity were measured. The fish grew throughout the experimental period, with the fastest rate during the first half of the first year of life. The highest mortality occurred during the first three months of the experiment and during the spawning season. The sex ratio was almost 1:1 (female:male). First sexual maturity was reached at one year of age, with an average weight of 0.85 ± 0.01 g, total length of 4.85 ± 0.16 cm, and condition factor 0.0074. The highest GSI in both females (12.14 ± 0.74) and males (17.7 ± 2.70) was recorded in August. Nevertheless, the females spawned 3 to 10 times between September and February, with the highest reproductive peak between September and October. The number of embryos per female per day varied from 1 to 429, while the total number of embryos per female during the entire season evaluated varied from 163 to 1044. There was a high correlation (r = 0.82) between absolute fecundity and body weight. Although further studies are needed in this field, these results are basic for establishing future reproductive programs in captivity as a strategy for sustainable fisheries and aquaculture management.

IVF Stimulation - personalized, optimized, and simplified using an advanced decision-support tool: A randomized trial

We assessed the effectiveness of a clinical decision support tool to reduce total cumulative follicle-stimulating hormone dosage used, to eliminate the need for ultrasound exams after day 5 of an individual superovulation cycle, and to improve the number of high-quality embryos obtained. The design we used was a Randomized Control Trial in a private fertility center. This study included 93 women aged 25–45 years undergoing IVF. 48 Test and 45 Control participants included normal and poor responders and patients with polycystic ovarian syndrome. A clinical decision support tool was used to forecast stimulatory hormone dosing for an individual cycle based on follicle size distribution on day 1 and day 5. Cumulative stimulatory hormone doses, oocytes retrieved, number of Mii oocytes, total embryos, high-quality embryos obtained during the cycle, and clinical pregnancy rates was the main outcome measures. Test participants required significantly lower cumulative FSH doses during superovulation cycles (average 1883 IU test, 2530 IU control, p &lt;0.01), with significantly higher numbers of total embryos (average 5.4 test, 3.5 control, p&lt;0.01), and high-quality embryos (average 3.1 test, 1.2 control, p&lt;0.01). Test participants had higher Mii follicles, although the difference was not statistically significant. The clinical pregnancy rate reported was significantly higher for test participants than control participants (52% test, 26% control, p&lt;0.05). The test group had more poor responders and PCOS patients than the control group. In conclusion, the clinical decision support tool eliminated the need for ultrasound exams after day 5, reduced the doses of stimulatory hormone required, yielded significantly higher numbers of high-quality embryos, and resulted in higher clinical pregnancy rates.

Dysregulation of TMEM16A impairs oviductal transport of embryos.

Ectopic pregnancy is an acute abdominalgia in obstetrics and gynecology, especially in fallopian tubal pregnancy. The ion channel protein transmembrane protein 16A (TMEM16A) is widely distributed in various tissues, even in the oviduct. In this study, we showed that TMEM16A was expressed in the human fallopian tube and was upregulated in patients with tubal pregnancy. By measuring isolated fallopian tube tissues, we found that TMEM16A was involved in regulating not only the contraction of muscle strips but also the beat frequency of cilia. In addition, pharmacological activation or inhibition of TMEM16A could lead to retention of embryos in oviducts. Moreover, the embryos in oviducts were delayed in development and some of them had malformations and deletions. The total number of embryos in the oviducts and uterus was significantly less than that of the control group. Furthermore, we detected changes in the level of m6A methylation, where the relevant writers and readers were reduced in tubal tissues from tubal pregnancies. In m6A mRNA methylation, writers catalyze the addition of methyl groups to cytosine residues and readers bind to the methyl groups and affect gene translation. In human fallopian tube epithelial cell line FTE187, we found that interference with methyltransferase 3 (METTL3) expression increased TMEM16A, suggesting that TMEM16A might be regulated by m6A methylation. In general, our study revealed a novel regulatory point for embryo transport and development, introducing a new role for the diagnosis and treatment of tubal pregnancy.NEW & NOTEWORTHY The ion channel protein TMEM16A is expressed in the epithelium and smooth muscle of the human fallopian tube and is upregulated in patients with tubal pregnancy. TMEM16A is involved in regulating the smooth muscle contraction and the cilia beating. Dysregulated TMEM16A may result in embryo retention in the oviduct and delayed early embryo development. Our study reveals a new regulatory point for embryo transport and development.

O-160 Do day of vitrification, grade of expansion and blastocyst quality affect embryo survival rate? An observational study in oocyte recipient patients

Abstract Study question Which parameters are the most predictive of embryo survival rate in oocyte recipient cryotransfer cycles? Summary answer We have identified that only day of vitrification and grade of expansion have an impact on embryo survival rate What is known already Embryo vitrification is a well stablished technique that has led to survival rates around ±96%. Recent studies, based on day of vitrification, grade of expansion and blastocyst quality, are trying to identify which vitrified blastocyst has the highest potential to survive and to implant. However, controversial results are present in the literature, improvements in laboratory quality parameters and embryo culture could unleash into no significant differences regardless of morphological parameters. Nevertheless, having an acceptable survival rate does not mean to have a good implantation rate, biomarkers to identify the best potential blastocyst to survive and to implant are needed Study design, size, duration An unicenter-based retrospective observational clinical study was performed at IVI Madrid, approved by the local institutional review board (IRB). The study included all the warmings at blastocyst stage from oocyte recipient patients from January 2019 to April 2022. A total of 4.196 of warmed blastocysts were assessed, out of which 4.006 blastocysts were transferred Participants/materials, setting, methods All warmed blastocysts were allocated in groups according to the day of vitrification, grade of expansion and quality. The primary outcome analysed was embryo survival rate that was calculated as the number of viable embryos divided by the total number of embryos warmed. Vitrification and warming were performed following Kitazato method (Cryotop, Kitazato, Biopharma). Embryo classification was assessed previous vitrification according to IVIRMA criteria. Statistical analyses were conducted using univariate and multivariate regression model Main results and the role of chance The mean maternal oocyte recipient age was 42.27±3.89 years old and body mass index 23±3.74 kg/m2). In a multivariate logistic regression analysis, a statistically significant association between embryo survival rate and the day of vitrification and grade of expansion was found. Chance of survival was significantly reduced for blastocyst vitrified on day 6 (n = 737) compared to day 5 (n = 3459) (93,2 % versus 96,0% [OR = 0.976 (0.960-0.993); p=0.05]). Regarding grade of expansion, cavitated blastocyst (n = 840) survival rate was 97,7% [OR = 1;p&amp;lt;0.001], expanded blastocyst (n = 1866) was 96,2% [OR = 0.095 (0.978-1.013)], hatching blastocysts (n = 1465) was 94% [OR = 0.978 (0.960-0.997); p=0.02) and fully hatched blastocysts (n = 25) was 60% [OR = 0.704 (0.648-0.765);p&amp;lt;0.001)]. However, no association was found neither between lower inner cell mass quality [OR = 0.964 (0.842-1.104); p=0.596) nor trophoectoderm [OR = 1.004 (0.981-1.028); p=0.746)] and survival rate. Analysing the combination of these three variables will give us the possibility to choose the best embryo to warm. The impact of day of vitrification, grade of expansion and blastocyst quality on implantation, clinical pregnancy, miscarriage, and live birth rates will be included in further studies Limitations, reasons for caution A weakness of this study is that it is retrospective in nature. The single-group design employed in retrospective studies limits the researchers’ ability to determine cause and effect. Further studies are needed to determine the impact of these variables on the survival rate Wider implications of the findings Based on our findings, blastocyst survival rate is influenced mostly by the day of vitrification and degree of expansion. To be able to select the best blastocyst to warm, a comparison between different day of vitrification when we have a similar morphology and embryo quality cohort is needed Trial registration number Not applicable

P-688 The relationship between higher levels of serum AMH and in vitro fertilization outcomes

Abstract Study question What is the relationship between increased serum anti-mullerian hormone (AMH) levels and in vitro fertilization (IVF) outcomes in women with infertility? Summary answer High levels of AMH are associated with higher oocytes and embryo number, but a lower proportion of GQE from the transferred and the total number. What is known already Although serum AMH level is considered a predictor of ovarian response to controlled ovarian stimulation, the predictive value of higher AMH levels is not clearly established due to contradictory reports, with both positive and negative associations with IVF outcomes. Study design, size, duration We performed a retrospective study in the Department of Reproductive Medicine of a private hospital. The medical records of all consecutive patients who underwent IVF between January 2014 and December 2021 with all causes of infertility were reviewed. The study group included 1401 patients with a median age [interquartile range] of 35 [6] years and median AMH of 1.99 [2.79] ng/mL. Participants/materials, setting, methods Patients with various causes of infertility undergoing IVF-ICSI and normal ovarian reserve (defined as a serum level of AMH above 1.1 ng/mL) were included in the study. Only patients with serum levels of AMH and age available for analysis were included in the study. The outcomes included were oocytes and obtained embryos number, the proportion of good-quality embryos from the total number of embryos obtained (GQE/T) and from the transferred number of embryos (GQE/Tr). Main results and the role of chance Patients were divided according to their serum AMH level into 3 groups: group 1 with AMH between 1.1 ng/ml and 5 ng/mL, group 2 with AMH between 5 and 8 ng/mL and group 3 with AMH more than 8 ng/mL. After adjustment for age, patients in groups 2 and 3 had a higher number of oocytes (beta 0.231, p &amp;lt; 0.0001 and beta 0.206, p &amp;lt; 0.0001) and embryos (beta 0.143, p &amp;lt; 0.0001 and beta 0.158, p &amp;lt; 0.0001). After adjustment for age, patients in group 2 had a lower proportion of GQE/T in comparison with patients in group 1 (beta -0.077, p 0.037) and patients in both groups 2 (beta -0.119, p 0.001) and 3 (beta -0.103, p 0.004) had a lower proportion of GQE/Tr in comparison with group 1. Being in groups with higher AMH (2 and 3) was not associated with the number of GQE. Limitations, reasons for caution Patients included in this study are infertile patients with an indication for IVF treatment. Therefore, the results of this study should be used with caution in other populations Wider implications of the findings Our study suggests that having higher levels of serum AMH (above 5 ng/mL) is not associated with better IVF outcomes in terms of the number of good-quality embryos. Moreover, obtaining a higher number of embryos is not associated with a proportional increase in good-quality embryo availability. Trial registration number NA