Artificial Insemination Research Articles

Laparoscopic artificial insemination of white-tailed deer with fresh sex-sorted semen

This case study describes the feasibility of using chilled, sex-sorted semen for laparoscopic artificial insemination (AI) in the captive white-tailed deer (Odocoileus virginianus) industry. Ejaculates from 3 bucks were collected, extended, and shipped overnight for flow cytometric sex-sorting. Sex-sorted semen was chilled and shipped back overnight. Synchronized does were bred using chilled, sex-sorted (n = 30, farm 1; n = 10, farm 2) or conventional frozen (n = 7, farm 2) semen by laparoscopic AI. Does were subsequently turned out with bucks. Percentage of bucks born via AI with sex-sorted semen was 85% (28/33) for farm 1 and 100% (16/16) for farm 2. Percentage of bucks born by natural cover was 50% (12/24) for farm 1 and percentage of bucks born by using either conventional semen or live cover was 56% (9/14) for farm 2. This case study demonstrated acceptable results using chilled, sex-sorted semen in white-tailed deer herds as a technique to manipulate the sex proportions per breeding season.

Synchronization of Ovulation and Fixed Time Breeding in Tellicherry Goats

Background: Reproductive management of goats on large scale becomes difficult due to contributing reasons like poor estrus expression, lack of heat detection techniques, etc. Synchronization of ovulation has significantly improved the conception in cows. However, such investigations are lacking in goats. Hence, an experiment was conducted to compare the efficacy of different ovulation synchronization protocols on kidding and fecundity rates in Tellicherry goats during winter and summer seasons. Methods: The study was conducted in 240 does of 60 days postpartum period in two different seasons and investigated the efficacy of different protocols, The efficacy of intravaginal sponge (group I), Ovsynch (group II), Ovsynch with sponge (group III), Co synch (group IV), Co synch with sponge (group V) and control (group VI) on kidding and fecundity rate following fixed time breeding (NS/AI) was investigated in Tellicherry goats. Result: The overall kidding rate recorded during winter season in natural service (NS) was 80.00 and Artificial Insemination (AI) it was 70.00 per cent and in summer season it was 60.00 and 55.00 per cent, respectively. During winter season the fecundity rates observed were 1.70, 1.60, 1.75, 1.60, 1.60 and 1.40 in group I, II, III, IV, V and VI, respectively. The corresponding values during summer season were 1.55, 1.40, 1.60, 1.45, 1.50 and 1.10, respectively. From this study, the ovsynch with intravaginal sponge protocol found to be an effective protocol for ovulation synchronization in Tellicherry goats.

Effects of rate of body weight gain during the first trimester of gestation on beef heifer and offspring performance, concentrations of hormones and metabolites, and response to vaccination.

Our study objectives were to evaluate the effects of divergent rates of body weight (BW) gain during early gestation in beef heifers on F0 performance, metabolic and endocrine status, colostrum immunoglobulins, and subsequent F1 calf characteristics, growth performance, concentrations of hormones and metabolites, and response to vaccination. Angus-based heifers (n = 100; BW = 369 ± 2.5kg) were adapted to individual feeding for 14 d and bred using artificial insemination with female-sexed semen. Heifers were ranked by BW assigned to either a basal diet targeting 0.28kg/d gain (LG, n = 50) or the basal diet plus an energy/protein supplement targeting 0.79kg/d gain (MG, n = 50) until d 84 of gestation. Dam BW and blood samples were collected at 6 time points during gestation; body composition was evaluated at d -10 and 84; and fetal measurements were taken on d 42, 63, and 84. At calving (LG, n = 23; MG, n = 23), dam and calf BW were recorded; and colostrum, calf body measurements, and blood samples were collected. Cow-calf pairs were managed on a common diet from calving to weaning, followed by a common postnatal development period for all F1 female offspring. Growth performance, hormone and metabolite profiles, feeding behavior, and reproductive performance were assessed from birth to pre-breeding in F1 heifers. Offspring were vaccinated against respiratory disease and bovine viral diarrhea pathogens on d 62.3 ± 4.13 and 220.3 ± 4.13 post-calving. By design, MG dams were heavier (P < 0.0001) than LG at d 84, and the BW advantage persisted until subsequent weaning of F1 calves. Concentrations of serum IGF-1 and glucose were increased throughout gestation (P < 0.001) for MG dams, whereas concentrations of NEFA were decreased (P < 0.001) in LG dams. Calves from MG dams were 2.14kg heavier (P = 0.03) and had larger chest circumference (P = 0.04) at birth compared with LG cohorts. Heifers from MG dams continued to have greater (P ≤ 0.03) BW gain and feed efficiency during the development period, but no differences were observed (P ≥ 0.13) in body composition, concentrations of hormones and metabolites, feeding behavior, puberty attainment, and response to vaccination in F1 offspring. Hence, early gestation rate of gain impacted BW and concentrations of glucose and IGF-1 throughout gestation in the F0 dam, resulting in altered F1 calf BW and measurements at birth and increased gain and efficiency during the development period.

First-line infertility treatment in normal or subnormal sperm: Interest of a simplified pre-IMSI test.

In the field of male infertility, when sperm is normal/subnormal, a few "add-on" routine tests can complete the basic semen examination. The aim of this study was to develop and evaluate a faster, simplified motile sperm organelle morphology examination (MSOME) technique for selected infertile patients with apparently normal/subnormal sperm and, in their background: failure of two or three intrauterine insemination (IUI) cycles, repeatedly fragmented embryos, embryonic development to blastocyst-stage failures, repeated miscarriages, a long period of infertility or 2 or more IVF attempts without pregnancy. Our test results were correlated with IUI, conventional in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI) and intracytoplasmic morphologically selected sperm injection (IMSI) outcomes. We validated an adapted version of the MSOME analysis called the pre-IMSI test (PIT), based on vacuole evaluation alone. 248 infertile patients from our assisted reproductive technology (ART) Center were retrospectively selected and split into three PIT score subgroups (patients with ≤8% (score I), 9 to 15% (score II) and ≥16% normal spermatozoa (score III)) based on the correlation between PIT results and each ART technique outcome. The choice of one or another of these ART techniques had been made according to the usual clinico-biological criteria. Clinical outcomes for each of the three PIT subgroups were compared individually for the different ART techniques. For ICSI, the effect of the PIT score subgroup was significant for clinical pregnancies (p = 0.0054) and presented a trend for live births (p = 0.0614). Miscarriage rates of IVF attempts were statistically different depending on the PIT score (p = 0.0348). Furthermore, the odds ratios of clinical pregnancy rates were significantly different according to PIT score subgroup when comparing ICSI vs. IMSI or IVF vs. ICSI attempts. IMSI appears to be recommended when sperm belongs to PIT score I, ICSI when it belongs to PIT score II and IVF or IUI when sperm is of PIT score III quality in selected infertile couples. The lack of statistical power in these PIT subgroups means that we must remain cautious in interpreting results. Our results support the interest of this simplified test for certain couples with normal/subnormal sperm to help choose the most efficient ART technique, even as first-line treatment.

Carboxyfullerene C60 preserves porcine sperm by enhancing antioxidant capacity and inhibiting apoptosis and harmful bacteria.

This study used a porcine model to systematically investigate whether carboxyfullerene C60 (CF-C60) can be used for the sperm preservation. The results indicated that CF-C60 supplementation can preserve porcine sperm quality during storage at 17 °C. This effect was attributable to improvement in the antioxidant capacity of sperm through a decrease in the reactive oxygen species (ROS) level. Additionally, CF-C60 can maintain mitochondrial function, inhibit sperm apoptosis through the ROS/Cytochrome C (Cyt C)/Caspase 3 signaling pathway, and mediate suppression of bacterial growth through the effects of ROS. Finally, the results of artificial insemination (AI) experiments indicated that insemination with CF-C60-treated sperm can increase the total number of offspring born and reduce the number of deformed piglets. Thus, CF-C60 is safe for use as a component of semen diluent for sperm storage.

Artificial insemination of mud crab, Scylla olivacea

Artificial insemination of mud crab, Scylla olivacea

Unlocking the potential of propolis: Concentration-dependent effects on Ossimi Ram (Ovis aries) semen quality

Reproductive biotechnology requires successful and efficient cryopreservation of semen since it is a crucial technique for artificial insemination, which preserves and multiplies the superior genetic origins of animals. Propolis, a natural antioxidant produced by honeybees, was examined in this study. Tris-citric-glucose-soybean lecithin extender supplemented with propolis ethanolic extract (P, μg/mL) at 0 (Control), 25 (T1), 50 (T2), 100 (T3), and 200 (T4) was used. The extended semen, prepared in a 1:20 ratio (semen: extender) at 37 °C, was equilibrated for 2 hours at 5 °C and packaged in straws (0.25 ml) before storage in liquid nitrogen at -196 °C. Visual characteristics (e.g., motility, livability, abnormality), DNA integrity, lipid peroxidation, and total antioxidant capacity of semen after thawing with P extract were thoroughly investigated. The results revealed that the low concentration (T1, 25 and T2, 50 µg/ml P) decreased lipid peroxidation, apoptotic sperm, improved sperm characteristics, and maintained the highest sperm cell viability. This resulted in improved reproductive performance and DNA integrity during cryopreservation of Ossimi ram semen. However, the high concentration (T3, 100 and T4, 200 µg/ml P) had the opposite effect and showed a low impact on maintaining cryopreserved Ossimi ram semen. In conclusion, Propolis shows potential for enhancing cryopreservation efficiency, but an optimal dosage is crucial to avoid adverse effects.

The timing of pronuclear transfer critically affects the developmental competence and quality of embryos.

Pronuclear transfer has been successfully used in human assisted reproduction to suppress the adverse effects of a defective oocyte cytoplasm or to bypass an idiopathic developmental arrest. However, the effects of the initial parental genome remodelling in a defective cytoplasm on the subsequent development after pronucleus transfer have not been systematically studied. By performing pronuclear transfer in pre-replication and post-replication mouse embryos, we show that the timing of the procedure plays a critical role. Although apparently morphologically normal blastocysts were obtained in both pre- and post-replication pronuclear transfer groups, post-replication pronuclear transfer led to a decrease in developmental competence and profound changes in embryonic gene expression. By inhibiting the replication in the abnormal cytoplasm before pronuclear transfer into a healthy cytoplasm, the developmental potential of embryos could be largely be restored. This shows that the conditions under which the first embryonic replication occurs strongly influence developmental potential. Although pronuclear transfer is the method of choice for mitigating the impact of a faulty oocyte cytoplasm on early development, our results show that the timing of this intervention should be restricted to the pre-replication phase.

Transcriptomics reveals that NAD(P)H affects the development of the Zig-zag eel (Mastacembelus armatus ♀) × Spiny eel (Sinobdella sinensis ♂) hybrid offspring leading to low hatching rates

Zig-zag eel (Mastacembelus armatus (2 n = 48)) and Spiny eel (Sinobdella sinensis (2 n = 48)) are two species of the Mastacembelidae family commonly found in southern China. Hybridization between the two has a very high deformity rate and a very low hatching rate. In order to investigate the reasons for this, the first hybridization between M. armatus and S. sinensis was carried out using artificial insemination, and the embryonic development of the hybrid offspring was examined using microphotography, and the malformations of the hybrid offspring were investigated by transcriptomics. The experiments showed that the average egg production was 4265.7 ± 322.94 (Mean ± SD), the average fertilization rate of hybrid offspring was 98.67 ± 0.58 % (Mean ± SD), the hatching rate was 12.06 ± 3.44 % (Mean ± SD), the deformity rate was 98.15 ± 3.21 % (Mean ± SD), and the embryonic development successively went through the five main stages of fertilized egg, egg cleavage, embryo formation, organogenesis, and exertion of membranes. Transcriptomics showed that the expression of NAD(P)H-related enzyme activity DEGs was increased, and many DEGs related to cell signaling molecule transmission and metabolic regulation are enriched in KEGG pathways, such as IL-17 signaling pathway, Osteoclast differentiation, TNF signaling pathway and MAPK signaling pathway. etc. The major types of DEGs corresponded to those coding for proteins. This study suggests that the high malformation rate in hybrid offspring may be caused by impaired synthesis of proteins during embryonic development.

Anti-Müllerian hormone does not predict cumulative pregnancy rate in non-infertile women following four IUI cycles with donor sperm.

To evaluate the predictive value of serum AMH for clinical pregnancy in non-infertile population undergoing intrauterine insemination with donor sperm (ds-IUI). This multicenter prospective study (ClinicalTrials.gov ID: NCT06263192) recruited all non-infertile women undergoing ds-IUI from June 2020 to December 2022 in three different fertility clinics in Spain and Chile. Indications for ds-IUI included severe oligoasthenoteratozoospermia, female partner, or single status. Clinical pregnancy rates were compared between women with AMH ≥ 1.1 and < 1.1ng/mL. The main outcome measure was the cumulative clinical pregnancy rate after up to 4 ds-IUI cycles. A total of 458 ds-IUI cycles were performed among 245 patients, of whom 108 (44.08%) achieved clinical pregnancy within 4 cycles, 60.2% of these occurring in the first attempt and 84.2% after two attempts. We found no significant differences in AMH levels or other parameters (such as age, BMI, FSH, AFC) between women who became pregnant and those who did not. Cumulative pregnancy rates and logistic regression analysis revealed that AMH ≥ 1.1ng/mL was not predictive of ds-IUI success. While a high positive correlation was observed between AFC and AMH (r = 0.67, p < 0.001), ROC curve analyses indicated that neither of these ovarian reserve markers accurately forecasts cumulative ds-IUI outcomes in non-infertile women. The findings of this multicenter study suggest that AMH is not a reliable predictor of pregnancy in non-infertile women undergoing ds-IUI. Even women with low AMH levels can achieve successful pregnancy outcomes, supporting the notion that diminished ovarian reserve should not restrict access to ds-IUI treatments in eligible non-infertile women.

Retrospective Review of Congenital Anomaly Cases in Ruminants

This study was designed to provide data on the incidence rates of congenital anomalies, their distribution by organs and systems in calves, lambs and kids brought to Van Yüzüncü Yıl University Surgery Clinic between 2017-2023. The animal material of the study consisted of 1104 ruminants, including 886 calves, 183 lambs and 35 kids, aged 0-6 months. In addition to clinical examination, anomalies were diagnosed by using direct and indirect radiographic examinations when necessary. In some cases, a definitive diagnosis was made by performing experimental laparotomy based on clinical and radiological examinations. 341 out of 1104 ruminants, including 234 calves, 96 lambs and 11 kids, suffered from congenital anomalies. Of the ruminants with congenital anomalies, 208 (61.00%) were male, 132 (38.70%) female and 1 (0.30%) hermaphrodite. Abdominal wall anomalies were determined with the highest number of 112 (32.9%) cases. These were followed by musculoskeletal system anomalies in 83 (24.4%) cases and gastrointestinal system anomalies in 56 (16.5%) cases. Head region anomalies occurred in 41 cases (12.1%), urinary system anomalies in 23 cases (6.8%) and CNS anomalies in 9 cases (2.6%). Multiple anomalies were recorded in 16 (4.7%) cases. In conclusion, congenital anomalies are frequently seen pathologies in ruminants in our country, especially in our region. Avoiding inbreeding, making artificial insemination widespread, improving the care and nutrition of the dam during pregnancy, correcting adverse environmental conditions and eliminating stress factors, avoiding over-the-counter medication misuse during pregnancy and most importantly informing field veterinarians and animal owners about the subject will contribute to the prevention of congenital malformations and losses in livestock.

Intrauterine Insemination vs Timed Intercourse in Letrozole and FSH-stimulated Cycles in Dysovulatory Infertility

Intrauterine Insemination vs Timed Intercourse in Letrozole and FSH-stimulated Cycles in Dysovulatory Infertility

Retraction notice to “Luteal phase support may improve pregnancy outcomes during intrauterine insemination cycles” [Eur. J. Obstet. Gynecol. Reprod. Biol. 157(1) (2011) 57–62]

Retraction notice to “Luteal phase support may improve pregnancy outcomes during intrauterine insemination cycles” [Eur. J. Obstet. Gynecol. Reprod. Biol. 157(1) (2011) 57–62]

Post-cooling sperm processing can rescue sperm quality of cooled-stored stallion semen

Poor sperm quality in cooled-shipped semen has been related to subpar fertility in horses. Therefore, this study aimed to evaluate the ability of post-cooling sperm processing to improve sperm parameters of cooled-stored stallion semen for artificial insemination. For all experiments, ejaculates were collected, processed, and diluted in skimmed milk-based (SM) medium and stored at 5 °C/24h. In all experiments an aliquot of unprocessed cooled semen was used as a control. In the first experiment (Exp 1.), cooled-stored semen from 16 stallions (n = 32) was processed by SpermFilter or centrifugation (600×g/10min) and resuspended in an egg yolk-based freezing medium containing permeating cryoprotectants (EY-C) for cryopreservation. Sperm recovery and motility parameters were immediately assessed after sperm resuspension in both groups and compared with unprocessed (Unp) samples. In Exp 2., cooled semen samples from six stallions (n = 18) were processed using SpermFilter and resuspended in SM or EY-C. Motility parameters and plasma membrane integrity were assessed in all groups (Unp, SM, and EY-C). In Exp 3, cooled semen from four stallions (n = 20) was processed by SpermFilter, resuspended in SM, EY-C, or egg yolk-based medium without cryoprotectants (EY-nC); and submitted to a thermoresistance test (37 °C/3h). Motility parameters, plasma membrane integrity and stability, mitochondrial membrane potential, mitochondrial superoxide generation, and DNA fragmentation index were evaluated in all groups. Finally, in Exp 4, 39 estrous cycles of 11 mares were inseminated with unprocessed (n = 6) cooled-stored semen or semen cooled at 5 °C/24h and then processed by SpermFilter and resuspended in SM (n = 5), EY-C (n = 11), EY-nC (n = 11), or centrifuged and resuspended in EY-C (n = 6). Overall, semen processing and resuspension in EY mediums (EY-C and EY-nC) improved sperm parameters compared with those of unprocessed semen (P < 0.05). Centrifugation (91 ± 5 %) recovered more sperm than SpermFilter (84 ± 9 %; P < 0.05). Sperm resuspended in EY-nC maintained better sperm parameters throughout the thermoresistance test than those in the other groups (P < 0.05). The fertility rates were similar between all groups (P > 0.05). In conclusion, processing and resuspension in EY medium can improve sperm parameters in post-cooled-stored stallion semen.

Characterizing the consistency of motion of spermatozoa through nanoscale motion tracing

To demonstrate nanoscale motion tracing of spermatozoa and present analysis of the motion traces to characterize the consistency of motion of spermatozoa as a complement to progressive motility analysis. Anonymized sperm samples videographed under quantitative phase microscope, followed by generating and analyzing super-resolution motion traces of individual spermatozoa. Centrifuged human sperm samples. Precision of motion trace of individual sperms, presence of helical pattern in the motion trace, and mean and standard deviations of helical periods and radii of sperm motion traces, speed of progression. Spatially sensitive quantitative phase imaging with a super-resolution computational technique MUltiple SIgnal Classification ALgorithm (MUSICAL) allowed achieving motion precision of 340 nm using 10×, 0.25 NA lens whereas the diffraction limited resolution at this setting was 1320 nm. The motion traces thus derived facilitated new kinematic features of sperm, namely the statistics of helix period and radii per sperm. Through the analysis, 47 sperms with speed >25 μm/sec randomly selected from the same healthy donor's semen sample, it is seen that the kinematic features did not correlate with the speed of the sperms. Also, it is noted that spermatozoa may experience changes in the periodicity and radius of the helical path over time. Further, some very fast sperms (for example >70 μm/sec) may demonstrate irregular motion, needs further investigation. Presented computational analysis can be used directly for sperm samples from both fertility patients with normal and abnormal sperm cell conditions. We note that MUSICAL is an image analysis technique which may vaguely fall under machine learning category, but the conventional metrics for reporting found in EQUATOR do not apply. Alternative suitable metrics are reported, and bias is avoided through random selection of regions for analysis. Detailed methods are included for reproducibility. Kinematic features derived from nanoscale motion traces of spermatozoa contain information complementary to the speed of the sperms, allowing further distinction among the progressively motile sperms. Some highly progressive spermatozoa may have irregular motion pattern, and whether irregularity of motion indicate poor quality regarding artificial insemination needs further investigation. Presented technique can be generalized for sperm analysis for a variety of fertility conditions.

Effect of altrenogest treatment before weaning on reproductive performance and production efficiency in primiparous and multiparous sows

BackgroundMost sows will experience negative energy balance during lactation resulting in impaired follicular development. This study aimed to treat 28-day lactating sows with altrenogest (ALT) to suppress follicle enlargement during lactation, and to assess the estrus and reproductive performance post-weaning.MethodsIn this study, we conducted two trials. In trial 1, we monitored the follicular development of lactating sows including 10 primiparous sows and 10 multiparous sows during the whole lactation to confirm the ALT administration time. In trial 2, a total of 42 primiparous and 111 multiparous sows were allocated to three treatments: Ctrl (control group, n = 51): no treatment; TAI (timed artificial insemination group, n = 51): sows were injected with equine chorionic gonadotropin (eCG) after weaning 24 h and gonadotropin-releasing hormone (GnRH) when they expressed estrus; and AT-TAI (ALT treatment-timed artificial insemination group, n = 51): base on the process of TAI group, the sows were fed with 20 mg ALT per day before weaning 10 days. All sows were artificially inseminated twice at 12 h and 36 h after estrus. The follicle size changes and serum hormone levels were explored in this process.ResultsAlthough the follicle size of multiparous sows was larger than primiparous sows during the whole lactation (P < 0.05), similar change trends of follicle size were observed in primiparous and multiparous sows. Meanwhile, the FSH, LH and E2 levels of multiparous sows were higher than primiparous sows. The ALT treatment significantly inhibits the increase in follicle size (P < 0.05) and reduces the serum levels of FSH, LH and E2 (P > 0.05). Additionally, ALT treatment increases estrus concentration and the preovulatory follicle size (P < 0.05), meanwhile, it delays the weaning-to-estrus interval (WEI, P < 0.001). However, the estrus rate, pregnancy rate, total pigs born and born alive did not differ between treatments (P > 0.05).ConclusionsThere were significant differences in the size of follicles in the lactation between primiparous and multiparous sows. ALT treatment during the last ten days of lactation concentrated estrus expression leading to higher work efficiency of breeder in batch production, however, with no improvement in reproductive performance.

P-782 Are pregnancy complications associated with the method of conception in female same-sex couples? Results of a retrospective, matched cohort study comparing ROPA and IUI

Abstract Study question Are pregnancies in female same-sex couples at a higher risk when using reception of oocytes from partner (ROPA) compared to those following intrauterine insemination (IUI)? Summary answer In female same-sex couples, pregnancies following ROPA demonstrated a higher incidence of preterm birth and first-trimester bleeding. What is known already Reception of Oocytes from PArtner (ROPA) enables both partners to contribute biologically to the desired pregnancy. However, oocyte donation, often performed in case of (premature) ovarian failure, is associated with an increased risk of hypertensive disorders of pregnancy/preeclampsia. Female same-sex couples typically comprising healthy, young women without underlying subfertility problems, are a priori at low absolute risk for pregnancy complications. Few studies have investigated the incidence of pregnancy complications and/or obstetrical outcomes in this specific group of women. Further insights and formal data could guide clinicians and patients in making informed decisions regarding the preferred method of conception. Study design, size, duration A retrospective, single-center, matched cohort study was performed including 577 unique female same-sex couples using donor semen who became pregnant after reproductive treatment (88 pregnancies following ROPA and 489 following IUI). None of the patients who opted for ROPA had (failed) IUI before. The study covered the period from January 2015 to January 2023. Participants/materials, setting, methods Pregnancies following ROPA were matched with pregnancies following IUI for age (+/-4 years), BMI (+/-4 kg/m2), parity (nulliparous vs multiparous) and date of embryo transfer/IUI (+/-1year) in a 1:1 ratio. Statistical Package for Social Sciences (SPSS) v26.0 was used for statistical analysis. Kolmogorov-Smirnov test was used to evaluate the normal distribution of continuous variables. Normally distributed data were analyzed with the student t-test and categorical variables were analyzed with the chi-square test. Main results and the role of chance After matching, 154 pregnancies (77 ROPA versus 77 IUI) were eligible for the study. Age (mean age 31.4±4.0 for ROPA versus 31.2±4.3 years for IUI, p = 0.48) and BMI (24.5±3.9 kg/m2 versus 25.2±4.4 kg/m2, for respectively ROPA versus IUI, p = 0.32) were comparable between groups after matching. Two ROPA pregnancies were excluded from further analysis due to elective termination of pregnancy (radiusaplasia and trisomy 21). Both groups experienced two stillbirths (two unexplained mors in utero, one chorioamnionitis and one intrapartum death). No significant difference in hypertensive disorders of pregnancy (HDP) (10.7% (8/75) versus 13.7% (10/73) for ROPA and IUI respectively, p = 0.53) or preeclampsia rates (11.0% (8/73) and 6.7% (5/75) for ROPA and IUI respectively, p = 0.33) was observed. ROPA-associated preeclampsia cases were often preterm (&amp;lt;37weeks’ gestation) compared to IUI (5/8 versus 1/5). ROPA pregnancies had a higher incidence of preterm birth (15.1% vs 1.3%,p&amp;lt;0.002) and first trimester bleeding was significantly more frequent (12.3% vs 0%,p&amp;lt;0.001). No statistically significant differences were observed between groups in neonatal weight, fetal structural malformations and cesarean section delivery rates. Limitations, reasons for caution The study is limited by its retrospective design and inherent bias. Despite reducing the sample size, matching enhanced study robustness. Recognized risk factors for hypertensive disorder of pregnancy, like the use of donor sperm, may mitigate differences associated with conception methods in this population. Wider implications of the findings The study points out potentially increased obstetrical risks following ROPA. These data are crucial to counsel and guide female same-sex couples in their decision regarding the method of conception. The risk of hypertensive disorders of pregnancy, specifically preterm preeclampsia, associated with ROPA warrants further attention, urging larger prospective studies. Trial registration number not applicable

P-449 Association of corpus luteum activity in early pregnancy and live birth after assisted reproductive technology (ART) and intrauterine insemination (IUI)

Abstract Study question What is the incidence of low corpus luteum activity in early pregnancy, and how are endogenous progesterone levels and live birth achievement associated? Summary answer Frequently occurring low corpus luteum activity (progesterone-level) in early pregnancy after ART and IUI strongly predicts pregnancy demise, though not independent of trophoblast activity (hCG-level). What is known already Ovarian stimulation and human chorion gonadotropine (hCG) administration may induce a luteal phase defect via negative feedback on the hypothalamic-pituitary-ovarian (HPO) axis. Luteal phase support (LPS) is routinely administered with bio-identical progesterone or oral dydrogesterone (DYD). DYD has high oral availability, minimal interaction with the pituitary gland, and no cross-reactivity in enzyme-linked immunosorbent assay (ELISA) with progesterone. Therefore, utilizing DYD as LPS in IVF/ICSI or ovulatory IUI cycles provides a unique perspective to assess endocrine luteal activity. While various studies explored progesterone and hCG post-IVF or IUI, none focused solely on the corpus luteum’s sole endocrine function and its outcomes. Study design, size, duration This single-center, retrospective study (with institutional review board approval) investigated hCG and progesterone serum levels after IVF/ICSI cycles or IUI cycles supported with DYD as the sole LPS. The study aims to assess their predictive value, individually and in combination, for sustaining pregnancy. Data from 189 IVF/ICSI and 198 IUI early pregnancies, respectively, with positive hCG test between January 2017 and June 2022, was extracted from the patient management system at the University Fertility Center. Participants/materials, setting, methods Included were only first IVF/ICSI and IUI treatment-cycles with serum hCG value ≥ limit of detection (0.6 IU/l) between days 9 and 16 after fresh embryo transfer or IUI, respectively. Additionally to hCG, endogenous progesterone was determined and the relationship between these and pregnancy maintenance was investigated using logistic regression analysis. Inclusion criteria encompassed hCG-triggered cycles using DYD as the sole LPS, administered at 30 and 20 mg daily for IVF/ICSI and IUI therapy, respectively. Main results and the role of chance The 10th, 25th, 50th and 75th quantiles of serum progesterone are at 0.28, 3.75, 33.0 and 98.7 µg/l in IVF/ICSI cycles and 0.48, 2.70, 18.40 and 26.6 µg/l in IUI cycles. Women in the first quartile of progesterone levels had nearly exclusively pregnancy demise: live birth rate 3/47 (7%, 95% confidence interval (CI) 1-19%) and 0/47 (0%, 95% CI 0-7.5%) in IVF/ICSI and IUI cycles, respectively. In both IVF/ICSI and IUI, progesterone and hCG were highly correlated (correlation coefficient 0.83 in both cases). Accordingly, the association of serum hCG levels with live birth achievement was similar in quartile analyses. Using receiver operating characteristic (ROC) curves and Youden’s indices, the optimal cut-off values for predicting live birth were determined as follows: 16.49 µg/l (89 % sensitivity (SN) and 59 % specificity (SP)) for IVF/ICSI and 15.89 µg/l (90 % SN and 70 % SP) for IUI therapy. Logarithmic regression analysis also revealed a significant correlation between progesterone levels and live birth in IVF/ICSI and IUI therapy, with a p-value of &amp;lt; 0.001. Limitations, reasons for caution Low progesterone in early pregnancy may be a consequence or a cause for pregnancy demise. DYD, like any other progesterone for LPS, might adversely impact corpus luteum formation and maintenance through HPO axis interaction, especially in mono-ovulatory cycles or in older women with inherently higher risk of luteal phase defect. Wider implications of the findings Ovarian stimulation and hCG-administration may induce a luteal phase defect with insufficient luteal activity in early pregnancy, even up to luteolysis, with potential consequences for pregnancy maintenance and maternal/fetal health. Future treatment protocols may require exploring an optimal luteotropic state at pregnancy’s outset, necessitating the establishment of relevant markers. Trial registration number not applicable

P-051 Results of Intrauterine Insemination (IUI) with donor semen in women with poor ovarian reserve

Abstract Study question If we have a woman with a poor ovarian reserve, will she have a lower chance of pregnancy after a cycle of IUI? Summary answer A woman with a poor ovarian reserve has a lower chance of pregnancy, although in donor IUI we have an acceptable rate of pregnancy. What is known already Actually, the better predictor to define a poor ovarian reserve is a low antimullerian hormone (AMH). AMH is a well-known predictor of ovarian reserve, which allow to predict a response after an in vitro fecundation (IVF) cycle. Nevertheless, we also know that a low AMH does not mean to be not able to get pregnant. Few studies have compared different markers of ovarian reserve as predictors of pregnancy after an IUI cycle, and those which have done it have conflictive results. Maternal age seems to be strongly linked with the chance of pregnancy. Study design, size, duration We designed a retrospective cohort study with longitudinal follow-up upon the patients who undergo an IUI cycle in a public hospital of third level (Hospital Virgen del Rocio, Seville, Spain). We included 1059 cycles of IUI (383 with conjugal and 676 with donor semen) between 2015 and 2021. These cycles belong to 390 patients (64 with poor ovarian reserve and 326 with normal ovarian reserve). Participants/materials, setting, methods We collected data from clinical history which is completed during the visit of the patients. Cohort was stratified between patients with a low AMH (&amp;lt;1.1 ng/ml) and normal AMH (&amp;gt;1.1 ng/ml). We used this cut point following the definition of our regional guideline that includes when we can practice fertility treatments. Our main variable was pregnancy rate, defining pregnancy as an echography at 7 weeks with at least one embryo with cardiac activity. Main results and the role of chance When we compared pregnancy rate between normal ovarian reserve against poor ovarian reserve, we did not find statistically significant results in conjugal IUI independently that we compared by cycle (11.2% in normal ovarian reserve vs 3.8% in poor reserve (p = 0.2)) or by couple (27.97% in normal ovarian reserve vs 8.3% in poor ovarian reserve (p = 0.1)). We found differences with the same comparison in donor IUI by cycle (20.2% normal reserve vs 6.8% in poor reserve (p = 0.001)) and by couple (56.83% in normal ovarian reserve vs 21.15% in poor reserve (p = 0.001)). Nevertheless, pregnancy rate in IUI was similar to what we will get with an IVF cycle with only one oocyte. We compared pregnancy rate between normal ovarian reserve patients who undergo a conjugal IUI cycle, with low reserve patients who undergo a donor IUI cycle. When we compared by cycle normal ovarian reserve patients had 11.2% pregnancy rate, while poor ovarian reserve patients had a 6.8% (p = 0.1). When we compared by couple normal ovarian reserve patients had a pregnancy rate of 27.97% while poor ovarian reserve patients had a 21.15% (p = 0.3). Actually, our regional guide excludes poor ovarian reserve woman of an IVF cycle in the public system. Limitations, reasons for caution We had the proper limitations of an observational retrospective study. As we are a public center, we must follow our regional guide which defines whether we can do a treatment or not. Wider implications of the findings Patients with a poor ovarian reserve have a lower pregnancy rate during IUI with donor semen. Nevertheless, this rate could be acceptable especially when we cannot offer them an IVF cycle in public healthcare. Trial registration number Not applicable

O-017 Swim-up versus density gradients for sperm preparation in intrauterine insemination: a multi-centre randomised clinical trial

Abstract Study question What is the effectiveness of swim-up (SU) and density gradients (DG) for sperm preparation in infertile couples undergoing intrauterine insemination (IUI)? Summary answer In couples undergoing IUI, there is no significant difference in the treatment outcomes between SU and DG. What is known already Swim-up (SU) and density gradients (DG) are the most two commonly used techniques for sperm preparation in intrauterine insemination (IUI). The latest Cochrane review concluded that there is uncertainty about the effectiveness of SU versus DG and no studies reported on live birth. Study design, size, duration This open-label, multi-centre randomised clinical trial was conducted on 912 infertile couples at a tertiary IVF center, Ho Chi Minh City, Vietnam from August 2020 to June 2023. Couples were randomly asigned to two groups, SU and DG using computerized randomisation on the day of IUI with variable block size of 2, 4 or 6. Primary outcome was live birth rate. Participants/materials, setting, methods Infertile couples were eligible if aged ≥18, indicated for IUI with ≤2 IUI prior attempts and husbands’ sperm concentration ≥5x106/ml, progressive motility (PR) ≥32% and total PR sperm count &amp;gt;5x106 before sperm preparation (WHO 2010 criteria). Couples using frozen semen, or high viscosity semen were not eligible. Women had ovulation induction using hMG. Sperm preparation was performed within one hour after ejaculation. IUI was performed once at 36 to 40 hours after hCG trigger. Main results and the role of chance Between August 2020 and June 2023, 912 couples were randomly assigned to SU (n = 456) or DG (n = 456). Baseline characteristics were comparable between the two groups (mean female age of 31.3 ± 0.4 years; total sperm count: 86.2 [52.9;137.6] million; sperm motility: 44.8 ± 8.2%). Live birth occurred in 55 (12.1%) couples in the SU group and 71 (15.7%) couples in the DG group (RR 0.77; 95% CI 0.56-1.07). Ongoing pregnancy and multiple pregnancy were also comparable between the two groups. There were no statistically significant differences between the SU and DG groups with respect to the occurrence of pregnancy complications, obstetric and perinatal complications, preterm delivery, birth weight and neonatal complications. Pre-planned subgroup analysis showed that the live birth rate was not materially affected by the IUI indications, total motile sperm after preparation (&amp;lt;1, ≥1 and &amp;lt;5, ≥5 and &amp;lt;10 and ≥10 million). Limitations, reasons for caution The main limitation of the study was its open-label design, due to the nature of the interventions. Wider implications of the findings Our study suggests that in couples undergoing IUI with progressive motility (PR) ≥32% and total PR sperm count &amp;gt;5x106 before sperm preparation, SU or DG can both be used for sperm preparation, with a preference for density gradients. Trial registration number NCT04477356