Non-obstructive idiopathic azoospermia vs azoospermia with antecedents of cryptorchidism: ways and probabilities of becoming parents

Background Non-obstructive azoospermia (NOA) with history of cryptorchidism and idiopathic NOA are the most common forms of NOA without genetic aetiology. Of all patients with one of these two types of NOA, only a few will have a positive TEsticular Sperm Extraction (TESE). Of those with positive extraction followed by sperm freezing, not all will have a child after TESE-ICSI. What are the ways and probabilities of taking home a baby for patients with NOA and a history of cryptorchidism compared with patients with idiopathic NOA? Results Patients with idiopathic NOA or NOA and a history of cryptorchidism who underwent their first TESE were included. The patients were divided into two groups: Group 1 was composed of 125 patients with idiopathic NOA and Group 2 of 55 patients with NOA and a history of surgically treated cryptorchidism. Our results showed that more than half of the NOA patients succeeded in becoming parents. The main way to fulfil their plans for parenthood is to use sperm or embryo donation (72%) for men with idiopathic NOA, whereas the majority of men with NOA and a history of cryptorchidism had a child after TESE-ICSI (58.8%). Conclusions In our centre, before considering TESE for a patient with NOA, we explain systematically TESE-ICSI alternatives (sperm donation, embryo donation or adoption). As a result, the couple can consider each solution to become parents.

Informal Sperm Donation in Russia

Rising infertility across the globe has created a growing demand for assisted reproductive technologies (ART). In recent years, apart from sperm donation in formal settings such as fertility clinics, informal donation practices have emerged and spread across Russia. These reproductive donation practices have become possible due to the development of social networks and private online platforms. We conducted a pilot study (eleven semi-structured interviews) of the informal sperm donation in Russia and analyzed donor-recipient interactions, donors’ expectations and experiences of finding recipients online. We focus on donors' motivations and on the meanings, which donors invest in this practice that consumes significant resources on their part (medical tests and artificial insemination costs, travel and accommodation expenses, sometimes mutually agreed financial support of future offspring). We interpreted the practices that coalesced around informal donation from the perspective of symbolic interactionism, because it allowed us to showcase how actors reflected on and formulated the meanings of their actions in the absence of externally imposed rules (legal regulations, established moral conventions). Since informal donation practices do not fit into the traditional schemes of interpretation, such research requires the actors involved in informal donation either to create their own schemes or to modify the existing conceptual frames in creative ways. The study shows that informal donors do not only provide their genetic material but also spend time and invested considerable resources to ensure their procreation, including eventual financial support of the child. At the same time, these men are not interested in marital relations or paternal relations with their offspring. Thus, the informal sperm donors do not associate the parental project with traditional family and its values. We conclude that ART engendered a new phenomenon, which might be described as extramarital reproduction. Assisted reproduction outside marriage ­gains footing in Russia and requires more detailed further study.

P–021 An accurate and reliable screening for SARS-CoV–2 in human sperm samples by RT-PCR: A requirement to evaluate the viral contamination risk during SARS-CoV–2 pandemic

Study question How to ensure a reliable and accurate detection of SARS-CoV–2 in seminal plasma and spermatozoa fractions of human sperm samples? Summary answer This RT-PCR assay showed high sensibility, repeatability and reproducibility for SARS-CoV–2 detection in seminal plasma and spermatozoa fractions, with a detection limit of 17 genomes/reaction. What is known already SARS-CoV–2 pandemic brings numerous concerns, such as the safety of gametes for patients undergoing assisted reproductive technologies, fertility preservation or sperm donation. Transient viremia and expression of SARS-CoV–2 receptors in testis and accessory glands bring the question of the presence of the virus in sperm samples. Moreover, the contamination during sperm collection may be possible. The few available studies about this issue mostly showed the absence of SARS-CoV–2 detection in semen of COVID–19 patients, except one reported study. All these studies performed SARS-CoV–2 detection with RT-PCR assays approved for naso-pharyngeal swabs, without a process specifically validated for semen fractions. Study design, size, duration Method validation was conducted between July 2020 and January 2021. SARS-CoV–2 direct detection was performed according to the French Society of Microbiology guidelines (SFM). Repeatability (n = 6), reproducibility (n = 3), limit of quantification (n = 2) and of detection (n = 6) were evaluated in seminal plasma (SP) and spermatozoa samples isolated after density gradient centrifugation and cryopreserved. In addition, variability of the whole analytical method efficiency was evaluated in samples of men with normal (n = 6) or altered sperm parameters (n = 6). Participants/materials, setting, methods Samples were surplus semen obtained from men undergoing routine semen analysis after granting informed consent. Assays were performed on SP and frozen spermatozoa fractions. After automated RNA extraction (MGISP–960, MGI-Tech®), real-time RT-PCR was performed using the one-step multiplex TaqPath COVID–19 kit (ThermoFisher®) targeting three viral regions (ORF1, nucleocapsid-N and spike-S proteins). An exogenous internal control was added before RNA extraction. Positive samples and dilution ranges were prepared with a standard (SARS-CoV–2 inactivated virus, QnosticTM Randox®). Main results and the role of chance RT-PCR assay applied for human sperm samples has been previously validated and is routinely used for SARS-CoV–2 detection in naso-pharyngeal swabs. We evaluated the efficiency of RNA extraction and RT-PCR for SARS-CoV–2 detection in semen fractions. The qualitative and quantitative performance of the whole analytical method was validated with an accuracy profile for SP and spermatozoa fractions. Overall, for repeatability, the standard deviation (SD) of the cycle threshold (Ct) was lower than 0.40 for the strong positive sample and 0.50 for the low positive one. An exception was observed for the S target of the low positive SP samples (SD = 3) which was consistent with S being the less sensitive target of the assay. For reproducibility, SD of the Ct was lower than 0.30 for the strong positive sample and 0.80 for the low positive, except for the S target of the low positive (SD = 1.5). The linearity range was determined for N target, the most sensitive target of the RT-PCR assay. It layed between 5200 and 52 SARS-CoV–2 genomes/reaction. The limit of detection of the RT-PCR assay was 17 viral genomes/reaction. Equal efficiency of the assay was observed for SP and spermatozoa independently of semen parameters (normal and altered sperm parameters). Limitations, reasons for caution: Our detection method was validated for the whole process: RNA extraction (reagents and system), RT-PCR (reagents and thermocycler QuantStudio 5TM) and for both SP and frozen spermatozoa fractions. Variability might be observed with a different extraction system or a different type of biological sample. Wider implications of the findings: This validated RT-PCR assay enables accurate and reliable screening of SARS-CoV–2 in SP and spermatozoa fractions, mandatory to investigate the presence of the virus in semen samples of patients undergoing assisted reproductive techniques, fertility preservation or sperm donation, and to ensure viral safety in the cryobanking process during covid–19 pandemic. Trial registration number EudraCT 2020-A01409–30

P–093 The use of donor sperm improves post-ICSI live birth rates in advanced maternal age women

Study question Can the use of donor sperm improve post-ICSI live birth rate in advanced maternal age (AMA) patients? Summary answer The use of donor sperm increases post-ICSI live birth rate while substantially reducing abortion occurrence in AMA patients. What is known already Oocyte DNA repair capacity decreases with maternal age, when sperm DNA integrity is particularly important to avoid the transfer of gene truncations and de novo mutations to the zygote. Optimal DNA repair activity in the zygote requires paternal inheritance of 8-oxoguanine DNA glycosylase (OGG1), a rate-limiting enzyme in the base excision repair pathway. However, the involvement of paternal aging and sperm quality in the severe drop in fertility observed in AMA patients has not been addressed. While strategies to mitigate the impact of AMA on fertility have exclusively targeted oocyte quality, the sperm contribution in this scenario remains somehow neglected. Study design, size, duration Retrospective, multicentric, international study including 755 first ICSI cycles with patients’ own oocytes achieving a fresh ET between 2015 and 2019, 337 of which using normozoospermic partner semen and 418 using donor sperm. The association of sperm origin (partner vs. donor) with live birth was assessed by univariate/multivariate analysis in non-AMA (<37 years, n = 278) and AMA (≥37 years, n = 477) patients. ICSI outcomes were compared between partner and donor sperm in non-AMA and AMA patients. Participants/materials, setting, methods The study was conducted in 3 fertility clinics including 755 Caucasian patients aged 24 to 42 years. Univariate/multivariate analyses were performed to test the association of sperm origin with live birth; infertility factor, maternal age, oocyte yield and number of embryos transferred were included in the model as confounding variables. In addition, ICSI outcomes were compared between donor and partner sperm groups with the Chi-square (percentages) or with the Wilcoxon sum rank (continuous variables) tests. Main results and the role of chance The multivariate analysis revealed that the use of donor sperm was positively and independently associated with live birth occurrence in AMA [1.82 OR (1.08–3.07) 95% IC; p = 0.024], but not in non-AMA patients [1.53 (0.94–2.51); p = 0.090]. Maternal age [0.75 (0.64–0.87); p < 0.001], number of MII oocytes recovered [1.14 (1.05–1.23); p = 0.001] and number of embryos transferred [1.90 (1.27–2.86); p = 0.002] were also independently associated with live birth in AMA patients. Live birth and delivery rates were 70–75% higher, while miscarriage rate was less than half in donor sperm compared to partner sperm AMA cycles (LBR: 25.4% vs. 14.5%, p = 0.003; DR: 22.5% vs. 13.5%, p = 0.008; MR: 18.0% vs. 39.5%; p = 0.009). Implantation (17.4% vs. 13.5%; p = 0.075) and clinical pregnancy rates (27.5% vs. 22.3%; p = 0.121) did not significantly differ between sperm donation and partner sperm AMA cycles. Male age was substantially lower (23.6 ± 5.2 vs. 41.4 ± 5.0; p < 0.0001) and oocyte yield was higher (5.1 ± 3.1 vs. 4.3 ± 2.6; p < 0.0001) in sperm donation compared to partner sperm AMA cycles, while maternal age did not vary (39.8 ± 1.6 vs. 39.6 ± 1.7; p = 0.348). Limitations, reasons for caution This study is limited by its retrospective nature and by differences in patients’ profiles between sperm donation and homologous cycles, although this variation has been controlled for in the statistical analysis. Wider implications of the findings: The findings suggest that donor sperm can improve live birth rates by drastically reducing miscarriage occurrence in AMA patients. Therefore, the present results may influence AMA treatment decisions and, above all, contribute for AMA patients to achieve a healthy birth. Trial registration number Not applicable