Comparison of ICSI and Conventional IVF in Non-Male Factor Patients with Less Than Four Oocytes

Background: We aimed to analyse our clinical results for a particular subgroup of patients with poor ovarian response (POR) in order to clarify if lower number of oocytes is a drawback for proceeding to C-IVFMaterials and methods: In this retrospective study, patient files of all couples (#1733) who underwent oocyte retrieval between January 2017 and December 2019 were reviewed and 191 cases diagnosed with non-male factor infertility in which ≤3 cumulus-oocyte-complexes available for fertilisation were analysed. Exclusion criteria were: woman age>42, patients with a history of previous ART trial, prenatal genetic testing cycles and couples undergoing total cryopreservation for any indication. Three groups were constructed depending on the method of fertilisation and on semen quality as follows: IVF non-male factor (Group 1,n=77); ICSI non-male factor (Group 2, n=65); ICSI male factor- ICSI/MF n=49 according to WHO reference values. Main outcome parameters were: fertilisation rate, implantation rate and live birth rate. Results: Fertilisation rate per collected COC was significantly higher in group 1 compared to the other two groups (85,68%, 72,58%, 73,33% respectively, p=0,004). FR per inseminated oocyte also tended to be higher in group 1 but not reaching a statistically significant level. Both techniques yielded similar implantation rates (20,42%, 28,49%, 23,33% respectively, p=0,407) and live birth rates (26,8%, 30,6%, 31,1% respectively, p=0.643).Conclusions: In the presence of normal semen parameters, low egg number is not an indication to perform ICSI. The choice of fertilisation method should be based primarily on semen quality, in combination with the patient’s previous history regardless of the ovarian reserve.

Cyclic adenosine monophosphate modulator supplementation on in vitro maturation of bovine oocytes

Study evaluated the role of cAMP modulator (Forskolin and 3-isobutyl-1- methyl xanthine) supplementation on developmental competence of bovine oocytes. Cumulus oocyte complexes recovered from bovine ovaries of unknown reproductive status were used for the study. Oocytes retrieved by aspiration method were graded based on cumulus cell distribution and culture grade oocytes were selected for the study. A total of 414 culture grade oocytes were taken and divided into two groups. Group I constituted of 201 oocytes in which pre-maturation was carried out for a period of 2 h. In group II, 213 oocytes were selected in which normal maturation was carried out. Maturation was assessed after 24h of culture in CO2 incubator maintained at 38.5o C in 95 per cent humidified atmosphere of 5 per cent CO2 . Fertilisation was carried out using frozen thawed semen and the presumptive zygotes were then transferred to culture media and cleavage was assessed 48 h after insemination. A significantly higher maturation rate (p ?0.05) was observed in group I compared to group II (86.85 ± 1.19 vs 79.88 ± 2.67). There was a highly significant increase (p ?0.01) in cleavage rate in group I (65.92 ± 1.23) compared to group II (59.29 ± 1.50). A higher fertilisation rate was observed in group I (75.35 ± 1.19) than group II (71.88 ± 2.56). It could be concluded that pre-maturation with cAMP modulators improved the developmental competence of bovine oocytes.

Follicular GH and IGF1 Levels Are Associated With Oocyte Cohort Quality: A Pilot Study

IntroductionOocyte quality contributes to the development of an optimal embryo and thus a successful pregnancy. The objective of this study was to analyse the association between oocyte cohort quality and the follicular levels of growth hormone (GH), insulin-like growth factor 1 (IGF1), 25-hydroxy vitamin D (25OHD), thyroid-stimulating hormone (TSH), free triiodothyronine (fT3), free thyroxine (fT4) and antithyroid antibodies, as a function of intracytoplasmic sperm injection (ICSI) outcomes.Material and methodsWe conducted a prospective comparative pilot study from January 2013 to December 2017. 59 ICSI cycles constituted an abnormal oocyte cohort (n=34 cycles, in which more than 50% of oocytes presented at least one morphological abnormality) and a normal oocyte cohort (n=25 cycles, in which 50% or less of the oocytes presented at least one morphological abnormality). GH, IGF1, 25OHD, TSH, fT3, fT4 and antithyroid antibodies were measured in follicular fluid.ResultsThe fertilisation rate was lower in the abnormal oocyte cohort (65.5% vs. 80%, respectively, p=0.012). Oocytes’ proportion with at least one abnormality was 79.4% in the abnormal oocyte cohort and 29.0% in the normal oocyte cohort. The mean number of morphological abnormalities per oocyte was significantly higher in the abnormal oocyte cohort. The follicular levels of GH (4.98 vs. 2.75 mIU/L, respectively; p <0.01) and IGF1 (72.1 vs. 54.2 ng/mL, respectively; p=0.05) were higher in the normal oocyte cohort. There was no association with follicular levels of TSH, fT3, fT4, antithyroid antibodies, or 25OHD.ConclusionOocyte cohort quality appears to be associated with follicular levels of GH and IGF1.

Effects of Anti-TPO Antibodies on the Outcome of In Vitro Fertilization-Embryo Transfer Cycles

BACKGROUND The objectives of the study were to investigate whether the incidence of antithyroid antibodies (ATAs) is related to an adverse outcome in the in-vitro fertilization and embryo transfer (IVF-ET) in terms of quality of oocytes retrieved, fertilisation, cleavage, embryo quality and implantation rate (primary outcome) as compared to patients without anti-thyroid antibodies and also evaluate the association of ATAs with abortion rate, biochemical pregnancy and clinical pregnancy (secondary outcome). METHODS A total of 52 women having anti-thyroid peroxidase antibody (anti- TPO Ab) level ≥ 35 IU/mL (Cases) and 21 women having anti-TPO Ab level < 35 IU/mL (Controls) undergoing IVF / ICSI from Jan 2014 to Dec 2014 at Milann – The Fertility Centre, Bangalore, were retrospectively analysed. RESULTS There was no difference among the cases and controls for the primary outcome, viz. maturation rate, fertilisation rate, cleavage rate, implantation rate, and embryo quality. However, the abortion rate and biochemical pregnancy rate were significantly higher in the cases than in controls (P = 0.027 for each parameter). On the other hand, the clinical pregnancy rate was significantly lower among the cases (P = 0. 045). CONCLUSIONS There was no statistically significant difference in the primary outcome among the two groups. The secondary outcome of abortion rate and biochemical pregnancy rate was significantly higher in the cases than controls. The clinical pregnancy rate was significantly lower among the cases as compared to controls. Clinical Significance - Anti-TPO Ab acts as a prognostic factor and is disadvantageous to the pregnancy outcome following IVF-ET. KEY WORDS Abortion Rate, Clinical Pregnancy Rate, Fertilisation Rate, Implantation Rate, IVF/ICSI, Oocyte Maturation Rate

Evaluation of the quality of embryo based on morphology cultured at low oxygen concentration

Objectives: evaluating the quality of embryo morphology cultured at low oxygen concentration (5%) at different stages of embryonic development: day 3, day 5. Methods: the present study examined 168 IVF/ICSI cycles from October 2019 to February 2021 at the Assisted Reproductive Center, 16A Ha Dong General Hospital. Embryos were randomly assigned to 2 groups: group 1 used a K-system G-210 tri-gas incubator (Australia) (5% O2, 5% CO2, 90% N2) while group 2 used a Thermo Scientific 371 dual-gas incubator (Denmark) (5% CO2, 75% N2 with 20% atmospheric Oxygen concentration). Participants in the study were patients younger than 37 years old, with AMH>1.2 ng/ml, and AFC≥4. Mature oocytes were injected with sperm, and cultured in a sequential medium (G1,G2-PLUSTM). Embryologists assessed embryos on the day of fertilisation, days 3, 5, and compared the results of the two groups, using the method of descriptive statistics and T-test. The results revealed an insignificant difference in fertilisation rate and the quality of cleavages cultured in these 2 groups (percentages of good- and average quality cleavages were, in turn, 77.28±4.62% và 77.99±5.03%, at p>0.05, number of poor quality cleavages were, in turn, 1.71±0.38 vs 1.97±0.49 with p>0.05). The results on day 5 embryo showed that the percentage of blastocysts (from fertilisation) and the percentage of morphologically good- and average-quality blastocysts tended to increase higher when cultured in 5% oxygen concentration (p<0.05) compared with the 20% one (57.79±3.60% and 53.05±4.50%, 78.62±4.42% and 70.97±5.67%, respectively). Conclusions: embryo cultured in a low oxygen concentration helps embryos develop better on day 5 than when cultured at atmospheric concentrations

P–756 Predictive factors influencing multiple live births in cumulative IVF cycles: retrospective analysis of over 265000 embryo transfer procedures from the national French registry

Study question What are the factors that could predict the number of embryos to be transferred in order to diminish risk of multiple pregnancies? Summary answer Single embryo transfer (SET) is advisable for &lt;38 year-old women in fresh cycles and for &lt;35 year-old women in FET whatever the IVF number attempts. What is known already Multiple pregnancies are associated to increased maternal and perinatal complications. Risks associated to multiple implantations are significantly reduced with SET policy. However, while SET is more assertive with a lesser negative impact in younger patients (&lt;35 years), its feasibility is less evident for the older population, whom oocyte quality is likely compromised. A double embryo transfer (DET) could improve chances of implantation and shorten their time to pregnancy. Identification of risk factors for multiple pregnancies could help in decision making for a double or SET and reduce chances for multiple gestations without reducing the chances to achieve pregnancy. Study design, size, duration A retrospective study from the national French data registry provided and approved by the Agence de la Biomédecine was performed. A total of 196530 fresh and 68913 frozen cycles from women aged 18–43 year-old were included (2014–2017). Risk factors assessed included women’s age, number of attempts, number of oocytes, fertilization rate, embryo stage, number of embryos transferred, number of supernumerary embryos frozen. Secondary infertility, oocyte donor, oocyte freezing, PGT, freeze-all and IVM cycles were excluded. Participants/materials, setting, methods Cumulative cycles derived from 65% of ICSI, 32% of IVF and 3,2% IVF/ICSI. The distribution of patients age at oocyte retrieval was 60% &lt; 35, 21% &lt; 38, 11% &lt; 40, and 8% ≥ 40 years old. Multivariable logistic regression was conducted to calculate adjusted odds ratios with 95% confidence intervals for live birth chance and multiple live birth risk associated with each risk factor. Main results and the role of chance The chances of obtaining a cumulative live birth decreases with increased patients age (OR 0.71 for 35–38 years and 0.47 for 38–40 years, p &lt; 0.00001), with increased number of attempts (from OR 0.87 for attempt = 2 to OR 0.74 for attempt ≥ 4, p &lt; 0.00001), and for frozen embryos transferred (OR 0.14, p &lt; 0.00001). The chances of live birth increases with the increased number of oocytes (from OR 1.33 for 4–12 to OR 1.52 for &gt; 18, p &lt; 0.00001 in all cases), with a fertilisation rate &gt;40% (OR 1.29, p &lt; 0.00001), with blastocyst transfer (OR 1.29, p &lt; 0.00001), with the increase on the number of frozen embryos (OR 7.37 for &gt;1, OR 13.08 for &gt;2, and OR 16.92 for &gt;6, p &lt; 0.00001 in all cases) and number of embryos transferred (OR 1.42 for 2 embryos and OR 1.39 for &gt;2 embryos, p &lt; 0.00001 in all cases). In case of live birth, the risks of multiple births when two embryos were transferred decreases in patients aged &gt;38 years (OR 0.50, p &lt; 0.00001) and for frozen embryos transferred (OR 0.65, p &lt; 0.00001). The risk increases with a fertilisation rate &gt;60% (OR 1.30, p &lt; 0.00001), with blastocysts transfer (OR 1.34, p &lt; 0.00001) and when at least one supernumerary embryo is frozen (OR &gt; 1.30, p &lt; 0.00001). Limitations, reasons for caution This study is limited in only providing a risk-benefit balance for multiples on the choice of transferring one or two embryos. Clinical data such as stimulation protocols and doses of gonadotropins were not considered in this evaluation. Wider implications of the findings: This study provides help to develop a strategy for the medical staff in the decision making for the number of embryos to be transferred. It may also serve as a patient’s information aid and help to improve their chances of achieving a health singleton if pregnant. Trial registration number Not applicable

P–001 Effect of mild α -chymotrypsin treatment of highly viscous semen samples on fertilization rates

Study question The main objective of this study was to examine the influence of mild α -chymotrypsin treatment of highly viscous semen samples on the fertilisation rate after artificial reproductive treatment (ART). Summary answer The use of mild α -chymotrypsin treatment of semen samples in case of hyperviscosity does not appear to impact negatively on the fertilization rates after ART. What is known already Highly viscous semen reduces sperm motility significantly and can contribute to infertility. When processing semen samples, few techniques exist to induce liquefaction in case of seminal hyperviscosity such as different washing steps and mechanical treatment. The use of α -chymotrypsin seems controversial due to possible negative effects on fertilisation rates after IVF. Study design, size, duration All patients were recruited at the Department of Gynecological Endocrinology and Reproductive Medicine, University Hospital of Bonn, Germany from July first 2018 till June first 2019. Anonymized data on age, number of previous pregnancies and deliveries were retrospectively collected. The study group consisted of a cohort of 52 couples meeting the inclusion criteria of highly viscous semen and were compared to a cohort control group of 88 couples. Participants/materials, setting, methods: The fertilization rate of 52 ART cycles was examined following IVF using a low dose of α -chymotrypsin to induce liquefaction of highly viscous semen and was compared to a control group of 88 ART cycles. Data was analyzed using SPSS version 25. A Mann Whitney U test was used to compare continuous parameters between groups. Main results and the role of chance The study group consisted of a cohort of 52 couples meeting the inclusion criteria of highly viscous semen and were compared to a cohort control group of 88 couples. The Fertilization rate between the two groups was not significant (p &lt; 0.146, Mann-Whitney U test), with a rate of 57.95 in the study group compared to 57.53 in the control group. Our analysis showed no significant differences in male and female age, male abstinence period, semen volume, sperm concentration, total sperm count, and total progressive sperm count between the two groups. We observed a significant difference [p = 0.025] in the progressive motility and a borderline significance in the total progressive sperm count [p = 0.052] between α -chymotrypsin treated samples and the non α -chymotrypsin treated samples. Analyzing the clinical and biochemical pregnancy parameters of the study group (n = 41) and the control group (n = 66) per fresh embryo transfer procedures (n = 107) we found no significant differences. Freeze all cycles were excluded from the study. No significant differences concerning previous pregnancies and childbirth were detected. Limitations, reasons for caution The study is a pilot study as the majority of the studies using α -chymotrypsin was conducted up to three decades ago, comparison of data must be interpreted in the light of the fact that since then, IVF has seen a rapid evolution of technology and culture techniques. Wider implications of the findings The use of mild α -chymotrypsin treatment of semen samples in case of hyperviscosity does not appear to impact negatively on the fertilization rates after ART and could be regarded as an additional method to induce liquefaction of highly viscous semen samples in IVF... Trial registration number not applicable

Evaluation of efficiency of controlled-release N fertiliser on tartary buckwheat production

To provide reference for scientific management of nitrogen (N) fertiliser on tartary buckwheat, the effects of the mixed application of controlled-release N fertiliser (a kind of thermoplastic polymer-coated urea types that are characterised by a semi-permeable membrane) and common urea was studied in the main tartary buckwheat production area in China. In 2018 and 2019, a two-year field experiment was conducted a randomised block design with five treatments: (1) no nitrogen fertilisation (CK); (2) 100% N from common urea (T1); (3) 15% N from controlled-released urea fertiliser (plastic coated) + 85% N from common urea (T2); (4) 30% N from controlled-released fertiliser + 70% N from urea (T3); (5) 45% N from controlled-released fertiliser + 55% N of urea (T4). The N fertilisation rate was 90 kg N/ha in all fertilisation treatments. The results showed: (1) the mixed application of controlled-release N fertiliser and common urea was conductive to enhance the yield, dry mass, N uptake and apparent N fertiliser efficiency (NFE), compared with a single application of common urea. In two seasons, NFE was 38.6% (T1), 48.6% (T2), 53.6% (T3) and 53% (T4), separately; (2) the mixed application of controlled-release N fertiliser and common urea could significantly increase the soil inorganic N content in the soil surface layer and decreased the leaching loss of N; (3) with the increasing ration of controlled-release N fertiliser, the tendency of increasing N content of crop uptake and soil residual and decreasing rate of N loss and N surplus was visible. Overall, considered the indicators of grain yield, input cost, N utilisation and N balance, the suitable N fertilisation mode for tartary buckwheat production is the mixed application of 30% controlled-release N fertiliser and 70% common urea when 90 kg N/ha is applied.

In vitro fertilisation (IVF) versus intracytoplasmic sperm injection (ICSI) in patients without severe male factor infertility: study protocol for the randomised, controlled, multicentre trial INVICSI

IntroductionOver the last decades, the use of intracytoplasmic sperm injection (ICSI) has increased, even among patients without male factor infertility. The increase has happened even though there is no evidence to support that ICSI results in higher live birth rates compared with conventional in vitro fertilisation (IVF) in cases with nonmale factor infertility. The lack of robust evidence on an advantage of using ICSI over conventional IVF in these patients is problematic since ICSI is more invasive, complex and requires additional resources, time and effort. Therefore, the primary objective of the IVF versus ICSI (INVICSI) study is to determine whether ICSI is superior to standard IVF in patients without severe male factor infertility. The primary outcome measure is first live birth from fresh and frozen-thawed transfers after one stimulated cycle. Secondary outcomes include fertilisation rate, ongoing pregnancy rate, birth weight and congenital anomalies.Methods and analysisThis is a two-armed, multicentre, randomised, controlled trial. In total, 824 couples/women with infertility without severe male factor will be recruited and allocated randomly into two groups (IVF or ICSI) in a 1:1 ratio. Participants will be randomised in variable block sizes and stratified by trial site and age. The main inclusion criteria are (1) no prior IVF/ICSI treatment, (2) male partner sperm with an expected count of minimum 2 million progressive motile spermatozoa following density gradient purification on the day of oocyte pick up and (3) age of the woman between 18 and 42 years.Ethics and disseminationThe study will be performed in accordance with the ethical principles in the Helsinki Declaration. The study is approved by the Scientific Ethical Committee of the Capital Region of Denmark. Study findings will be presented, irrespectively of results at international conferences and submitted for publication in peer-reviewed journals.Trial registration numberNCT04128904. Pre-results.