Groupe de Recherche en Fertilité Humaine

Numerous publications have questioned the lack of analytical reliability and clinical relevance of sperm morphology assessment for infertility workup and before use of assisted reproductive techniques (ART). There is a huge variability in the performance and interpretation of this test. It has become necessary to evaluate its true medical service rendered to the patient. To develop clinical guidelines for use of spermatozoa morphology assessment during male fertility check-up and before ART. These guidelines were produced following a pre-defined standard methodology for narrative and Patient Intervention Comparison Outcomes (PICO) questions. The French Working Group (WG) on Sperm Morphology Assessment consisted of 15 members including an expert in statistics. R1: WG does not recommend systematic detailed analysis of abnormalities (or groups of abnormalities) during sperm morphology assessment. R2: WG recommends that the laboratory should use a qualitative or quantitative method for detection of a monomorphic abnormality (globozoospermia, macrocephalic spermatozoa syndrome, pinhead spermatozoa syndrome, multiple flagellar abnormalities). The result may be given as an interpretative commentary or as a numerical report of the percentage of detailed abnormalities. R3: There is insufficient evidence to demonstrate the clinical value of indexes of multiple sperm defects (TZI, SDI, MAI) in investigation of infertility and before ART. Accordingly, the working group does not recommend the use of sperm abnormality indexes (TZI, SDI, MAI) in sperm morphology assessment. R4: WG gives a positive opinion on the use of automated systems based on cytological analysis after staining after qualification of the operators, and validation of the analytical performance within their own laboratory. R5: WG does not recommend using the percentage of spermatozoa with normal morphology as a prognostic criterion before IUI, IVF, or ICSI, or as a tool for selecting the ART procedure. This article examines the clinical interest of sperm morphology assessment during fertility check-up and before ART. The overall level of evidence from studies is low, challenging current practices regarding sperm morphology assessment. These guidelines suggest a significant simplification of sperm morphology assessment in the light of the examined publications while maintaining the detection of monomorphic sperm abnormalities.

Abstract Study question What is the impact of a commercially available antioxidants-enriched culture medium on blastocyst ploidy rates? Summary answer The rates of euploidy, aneuploidy, as well as of low and high mosaics are comparable to those achieved with a standard single-step culture medium. What is known already The in vitro manipulation of gametes and embryos is a significant stress factor associated with the generation of reactive oxygen species (ROS), which can accumulate over time during development. Oxidative stress (OS) occurs when ROS levels exceed the cellular antioxidant defence capacity, potentially compromising IVF outcomes by impairing gamete function and embryo development. Substantial evidence highlights the benefits of antioxidant supplementation in culture media, demonstrating improvements in key parameters such as blastocyst developmental rates in both animal models and humans. However, studies evaluating its impact on ploidy outcomes—a critical marker of embryo viability—are notably lacking. Study design, size, duration This is a single center ongoing randomized study with 1,091 MII oocytes in the present interim analysis obtained from 93 pre-implantation genetic testing for aneuploidy (PGT-A) cycles between March and December 2024. Participants/materials, setting, methods Patients included were 18–43-year-old and had ≥6 MII oocytes. Following denudation, MII oocytes were randomized into two culture media: antioxidants-containing medium, Gx (Vitrolife) or standard single-step medium, GT (Global-Total LP, CooperSurgical). ICSI was performed 1-hour post-denudation, and injected oocytes were placed into their respective medium, Gx or GT, for culture. Blastocysts graded ≥BL3CC on Days (D)5-7 underwent trophectoderm (TE) biopsy for next-generation sequencing (NGS) analysis. Post-insemination outcomes and blastocyst ploidy were assessed by mixed-effects-logistic-regression. Main results and the role of chance A total of 546 and 545 MIIs from Gx and GT medium were inseminated, respectively. Gx-medium was associated with a lower likelihood of achieving 2PN (OR:0.67; 95%CI: 0.50 – 0.90, p=0.008) (75.8% vs 82.1% 2PN in Gx vs GT, respectively), but no association was observed with abnormal fertilization (<3.0% of abnormal fertilisation for both) or oocyte degeneration. There were no significant differences observed in blastulation rates per 2PN for D5 (OR: 1.27 [0.83–1.95]; p= 0.263), D6 (OR: 1.10 [0.82–1.48]; p= 0.532), or D7 (OR: 1.98 [0.85–4.59]; p= 0.114). The use of Gx did not impact TE or inner cell mass (ICM) quality, neither blastocyst formation per injected MII (38.3% vs 36.9% in Gx vs GT, respectively). There was a borderline positive association between Gx-medium and total blastocyst formation per 2PN (OR: 1.32 [0.99–1.75]; p= 0.054) (50.6% vs 44.8% in Gx vs GT, respectively). Blastocyst euploidy per 2PN (OR: 1.23 [0.90–1.69]; p= 0.194) or per biopsy (OR: 1.03 [0.69–1.53] p= 0.900) were not impacted, neither the rates of mosaicism per 2PN (OR: 1.81 [0.43–7.64]; p= 0.417) or per biopsy (OR: 1.61 [0.38–6.82]; p= 0.519). Limitations, reasons for caution Oocytes were exposed to Gx-medium only 3 hours after COC retrieval, during which they had been placed in the standard single-step GT medium. Therefore, the effects of placing COCs immediately after retrieval into the antioxidant-containing Gx medium remained unexplored, warranting further investigation to clarify its potential benefits. Wider implications of the findings Gx-cultured oocytes while exhibiting lower fertilization rates delivered similar rates and quality of blastocysts in a cohort of MII oocytes, without impacting ploidy rates. Whether Gx-medium have an impact on implantation rates and on oocytes from other patient populations remains to be investigated. Trial registration number No

To develop and validate a predictive model for live birth (LB) outcomes in single euploid frozen embryo transfers (seFET) based on patient's characteristics and embryo parameters. A retrospective cohort study was performed including 1979 seFET performed between March 2017 and December 2023. Prediction models were built using logistic regression (LR), random forest classifier (RFC), support vector machines (SVM), and a gradient booster (XGBoost). Considered variables associated with LB outcomes were blastocyst expansion, blastocyst inner cell mass (ICM) and TE quality, day (D) of TE biopsy (D5, D6, and D7), female age and body mass index (BMI), distance from the uterine fundus at embryo transfer, endometrial preparation as natural cycles (NC) or hormonal replacement therapy (HRT), and endometrial thickness. Model performance was evaluated using area under the precision-recall curve and calibration metrics. Variables that were negatively associated with LB rate were BMI (OR = 0.79 [0.64-0.96], P = 0.020 for overweight and OR = 0.76 [0.60-0.95], P = 0.015 for obese class I/II), ICM grade B (OR = 0.72 [0.57-0.90], P = 0.005) or C (OR = 0.21 [0.15-0.30], P < 0.001), TE grade C (OR = 0.32 [0.24-0.43], P < 0.001), and blastocyst biopsied on D6 (OR = 0.66 [0.55-0.80], P < 0.001 or D7 (OR = 0.19[0.09-0.37], P < 0.001). The LR model was the best in terms of overall classification performance (C-statistics: 0.626 ± 0.018 vs. 0.606 ± 0.018, 0.581 ± 0.018, 0.601 ± 0.017, LR vs. RFC, XGBoost, and SVM, respectively, P < 0.001). A prediction model of LB outcome was developed and is free to access: https://artfertilityclinics.shinyapps.io/ABLE/ . LR demonstrated a stable validation performance and superior LB prediction, aiding as a predictive tool for patient counselling and assessing success in seFET cycles.

IntroductionA visual digital tool called SEXual Dysfunction Evaluation Tool (SEX-DET) was created for assessing the causes of sexual dysfunction, based on a biopsychosocialsexual and relational schema, to assist patients in understanding the relative influence of the various factors contributing to their sexual dysfunction. The aim of this study was to evaluate satisfaction of this tool in sexual dysfunction patients and practitioners. MethodsA satisfaction study of the SEX-DET was performed in 50 patients (20 F, 30 M) consulting for a sexual trouble and 3 practitioners. The main reasons for consultation were erectile dysfunction (36%) and desire disorder (30%). ResultsAll the participants agreed that the SEX-DET helped them to understand the origin their trouble (46% strongly, 42% moderately, 12% somewhat), the various factors involved (56% strongly, 22% moderately and 18% somewhat) and the management of their trouble (6% strongly, 47% moderately, 33% somewhat). Overall, 94% of participants found that the SEX-DET was easy to use (58% strongly, 16% moderately and 20% somewhat). Practitioners acknowledged the user-friendly interface, its critical role in improving patients’ understanding and providing a comprehensive visualization of the dimensions associated with sexual dysfunction, even if one practitioner threatened the tool's intrinsic inclination to oversimplify complex data. ConclusionSEX-DET enabled to improve patients’ understanding of their trouble and helped practitioners in structuring better communication during consultations communication regarding sexual dysfunction. The shortcomings of this study was its focus on short-term outcomes, self-reported data, and the absence of control group. Level of evidence3.

Severe acute respiratory syndrome coronavirus 2, the virus responsible for coronavirus disease 2019, affects multiple organs. The virus enters cells through angiotensin-converting enzyme-2 and host factors present in genital organs, leading to concern over virus shedding in semen and reproductive function. To investigate severe acute respiratory syndrome coronavirus 2 in semen from patients with a mild infection, identify the seminal infected cells, and explore the effect of the infection on sex hormones and semen parameters. Prospective study of 54 men with mild severe acute respiratory syndrome coronavirus 2 infection. Semen was collected at 7, 15, 30, 60, 90, 180, and 365 days after symptom onset, and severe acute respiratory syndrome coronavirus 2 RNA was measured in serum, saliva, urine, and semen. The presence of infectious severe acute respiratory syndrome coronavirus 2 in semen was assessed using Vero cell culture. Infected semen cells were identified using immunofluorescence against severe acute respiratory syndrome coronavirus 2 nucleoprotein antigen and cell markers. Semen characteristics as well as testosterone, inhibin B, luteinizing hormone, and follicle-stimulating hormone levels were determined. 11% of patients had at least one severe acute respiratory syndrome coronavirus 2 RNA-positive semen. One patient had viral semen shedding up to day 90 after infection onset, with replication-competent virus isolated from semen and 40% cell fraction at day 7. After sperm preparation, 90% fraction was severe acute respiratory syndrome coronavirus 2 RNA-positive at days 7 and 15. The swim-up fraction was positive only on day 7. In semen, nucleoprotein antigen was detected mainly in exfoliated epithelial cells and less frequently in Sertoli cells. Sperm count and motile sperm count were lower at day 30 than at day 7. Round cells in semen were increased during the acute phase. At days 7 and 15, sperm count and motile sperm count were lower in severe acute respiratory syndrome coronavirus 2 RNA-positive semen compared with negative semen, while semen volume and follicle-stimulating hormone levels were increased. Long-term follow-up shows no evidence of a detrimental effect on hormonal or semen characteristics. 11% of patients with mild coronavirus disease 2019 who were not hospitalized had severe acute respiratory syndrome coronavirus 2 excretions in semen, which persisted for up to 90 days in one patient. No germ cells appeared infected by the virus, but the detection of nucleoprotein antigen-positive epithelial semen cells and Sertoli cells suggests genital tract infection. Albeit infrequent, semen may contain the replication-competent virus during the acute phase with potential risk of severe acute respiratory syndrome coronavirus 2 transmissions during sexual contact and assisted reproduction procedures. The effect of mild coronavirus disease 2019 on spermatogenesis and reproductive hormones was moderate and reversible.

Purpose: The objective of this study was to evaluate the population density of quinoa plants sown by broadcasting or in rows to improve agronomic indicators of an organic farming system focused on increasing food and nutrition security of the population. Method/design/approach: The study was conducted in 2020, in Augusto Pestana, RS, Brazil, using a randomized block experimental design with four replications, in a 2×4 factorial arrangement consisted of 2 sowing methods (sowing in rows spaced 0.20 m apart and by hand broadcasting) and 4 plant population densities (10, 40, 80, and 120 plants m-2). Thinning was performed in the plots of both sowing methods approximately 20 days after emergence for achieving the desired plant densities. Grain yield and other traits of agronomic interest were evaluated under an organic cropping system. Results and conclusion: The arrangement of 70 quinoa plants m-2 through sowing in rows or by broadcasting ensures satisfactory crop yields without using pesticides and synthetic fertilizers. The incorporation of these more sustainable practices into quinoa production systems contributes to the promotion of agroecological and organic agriculture, resulting in a high-quality nutraceutical food. Originality/value: This study provides significant contribution to the advancement of research on quinoa in Brazil, bringing new information benefiting farmers, technical assistance agents, and the food industry and improving the production system efficiency to ensure food and nutrition security and care for the environment and society.

Abstract Study question Does cumulative livebirth-rate (CLBR) after reutilization of vitrified oocytes differ between patients having undergone fertility preservation (FP) for oncologic (onco-FP) reason or benign conditions (benign-FP)? Summary answer Although CLBR per woman was lower in the onco-FP group, there was no statistically significant association in multivariate analysis between onco-FP and CLBR. What is known already Recent ESHRE guidelines reported oocyte vitrification after controlled ovarian hyperstimulation (COH) as an established option for FP. Although age at the time of oocyte vitrification remains the main predictive factor of success, several lines of evidence suggest that the use of COH or in vitro maturation (IVM), as well as the indication of FP, in particular the type of disease, may influence outcomes after devitrification. The present investigation aimed at clarifying this issue. Study design, size, duration Observational comparative, monocentric retrospective study including all patients having reused, between January 2014 and December 2021, their oocytes vitrified for oncologic or benign conditions between 2013 and 2021. Women having undergone FP for non-medical indication were excluded. Participants/materials, setting, methods Among the 2201 patients having vitrified their oocytes, 94 (4.7%) returned for reutilization. The primary objective was the comparison of CLBR between onco-FP (n = 48) and benign-FP (n = 46) groups. A logistic model was performed. Factors associated with the CLBR in univariate analysis were included in a multivariate model. A secondary analysis was performed, comparing the benign-FP group and 2 groups of onco-FP according to the use of COH (Onco-COH, n = 25) or in-vitro maturation (Onco-IVM, n = 23). Main results and the role of chance Overall, in comparison with benign FP, women with malignant diseases were younger (median [IQR]: 34.0 y [31.0;37.0] vs. 36.5 y [33.2;38.0], p = 0.04) and had fewer oocytes vitrified (median [IQR]: 6.0 [3.0;9.2] vs.15.5 [6.2;18.0]), p &amp;lt; 0.001). The CLBR in onco-FP and benign-FP groups were 14.6% (7/48), and 32.6% (15/46), respectively. In univariate analysis, the CLBR was significantly lower in the onco-FP group when compared with the benign-FP group: (OR [95%CI]: 0.35 [0.12;0.94], p = 0.04). However, this difference in the CLBR did not reach significance after multivariate analysis (OR [95%CI]: 0.38 [0.09;1.51], p = 0.18). As expected, age at the time of oocyte vitrification was negatively associated with CLBR (OR [95%CI]: 0.80 [0.67;0.92], p = 0.005), while the number of oocytes inseminated was positively related to CLBR (OR [95%CI]: 1.22 [1.08;1.40], p = 0.002). Unlike onco-COH, onco-IVM was associated with a significant decrease in CLBR when compared to benign-FP (only COH), in univariate analysis: 8.7% (OR [95%CI]: 0.20 [0.03;0.80], p = 0.04), without significance after multivariate analysis (OR [95%CI]: 0.37 [0.04;2.16], p = 0.29).The livebirth rate (LBR) per oocyte was 2.1% in the onco-IVM subgroup (3 babies for 141 oocytes thawed), 2.7% in the onco-COH subgroup (7 babies for 260 oocytes thawed) and 3.7% (18 babies for 479 oocytes thawed) in the benign-FP group. Limitations, reasons for caution The small size of our population was probably responsible for a lack of statistical power. The retrospective and monocentric nature of our study may also be a weakness. Furthermore, LBR per oocyte was analyzed without age-adjustment. Wider implications of the findings This study provides actual data on chances of achieving a livebirth after oocyte vitrification, according to the use of a COH or IVM and the context of FP, oncologic or benign conditions. Trial registration number not applicable

Abstract Study question Does a short-time co-incubation in conventional IVF improve rates of fertilization, embryo development and euploidy, when compared to overnight co-incubation? Summary answer There is no significant difference in fertilization, embryo development and euploidy rates between sibling oocytes subjected to short co-incubation versus overnight co-incubation. What is known already Conventional IVF usually implies coincubation of cumulus-oocyte complex(es) with sperm for an overnight period. This overnight incubation might create an over exposure of COCs to reactive oxygen species (ROS), which might disturb oocyte homeostasis, and impact on embryonic developmental rate. To this extend, many early studies have compared a short co-incubation time (2-6 hours) with overnight co-incubation, with reported conflicting outcomes. None of the studies combined time-lapse monitoring with short co-incubation, which allows the identification of early morphokinetic events of the inseminated oocytes after conventional IVF, as already known for ICSI oocytes in time-lapse incubators. Study design, size, duration This single center prospective study included 604 COCs in 38 IVF/ICSI cycles with preimplantation genetic testing between December 2020 and November 2022. After oocyte retrieval a min of 6 COCs were assigned for insemination by IVF and equally divided between short(2hrs) and overnight (16-20hrs) co-incubation. Supernumerary oocytes were denuded and inseminated by ICSI. Primary endpoint was fertilization rate; secondary endpoints were maturation rate, ploidy rate after trophectoderm biopsy, embryo development and morphokinetics. Participants/materials, setting, methods Couples with a female age between 18-43 (average:34.4) years, BMI ≤30kg/m2 and normal semen parameters (WHO) in the fresh ejaculate on the day of OR, were eligible for the study. After IVF, all embryos were cultured in a time-lapse imaging system in Global Total LP media. Blastocysts (≥Bl3BCC) were subjected to trophectoderm biopsy on day 5-7 and next generation sequencing (NGS) to determine blastocyst ploidy status. Main results and the role of chance In total, 288 oocytes (47.7%) were denuded for ICSI, whereas 316 (52.3%) oocytes were randomized 1:1 to short and overnight co-incubation groups. Maturation rates (M2/COC) and normal fertilization rates (2PN/COC) in short and overnight co-incubation groups were similar (88.0% vs 83.5%, P = 0.074 and 60.8% vs 62.0%, P = 0.193, respectively). Day-3 embryo cell numbers (P = 0.383), fragmentation rates (P = 0.529), blastulation rates on day 5 (39.9 vs 38.0% P = 0.696), time to blastulation (median: 112.6 vs 111.1 hours after insemination by time lapse, P = 0.544) were similar between short and overnight co-incubation groups. In total, 48 embryos in short and 45 embryos in overnight co-incubation groups were of sufficient quality for biopsy resulting in similar euploidy rates between the groups (39.6 vs 44.4%, P = 0.791). When compared to their siblings in ICSI, short (mean difference: -6.48%, 95% CI: -18.5 to 5.59%, P = 0.290) and overnight co-incubation (mean difference: -0.87%, 95% CI: -12.9 to 11.2%, P = 0.886) attained similar fertilization rates per cycle. No significant differences regarding blastulation and euploidy rates were observed in short or overnight co-incubation compared to ICSI in unadjusted and adjusted analyses. Limitations, reasons for caution This is a prospective study with a limited patient population with a normal ovarian response. Results might not be transferable to patients with high or low ovarian reserve. Wider implications of the findings The present study demonstrated that there is no difference between short and overnight co-incubation in terms of fertilization and euploidy rates. Therefore short co-incubation might allow us to benefit from early denudation, and to investigate early morphokinetics in time lapse monitoring, which are otherwise missed by overnight co-incubation. Trial registration number NCT04627545

Simple SummaryTesticular cancer is the most common cancer among men below 40 years old, and its causes remain largely unknown. Marked geographic differences in the occurrence along with the relatively young age at diagnosis suggest a possible role of environmental and occupational exposures that occur early in life, probably already during embryonic development. Using data from a French case–control study, we investigated associations between paternal occupational exposure to five heavy metals and welding fumes before and at birth and testicular cancer risk in sons. We estimated exposures from job titles and considered other potential risk factors in the statistical analyses. We found no association. Thus, further research is necessary to identify potentially modifiable risk factors.Testicular cancer is the most common cancer in young men. Its causes are largely unknown, although prenatal occupational and environmental exposures have been suggested. We investigated paternal occupational exposure to heavy metals and welding fumes and the risk of testicular germ cell tumors (TGCT) in their offspring. A total of 454 cases and 670 controls were included from a French nationwide case–control study. The INTEROCC job exposure matrix was used to assign occupational exposures (cadmium, chromium, iron, nickel, lead, and welding fumes) to the fathers’ jobs. Odds ratios (ORs) for TGCT were estimated using conditional logistic regression models for frequency-matched sets. Three complementary analytical approaches were used: (1) single-agent analysis, (2) analysis by groups, and (3) principal component analysis (PCA). The proportion of paternal exposure to different heavy metals and welding fumes ranged from 0.7% (cadmium) to 11.3% (lead). Based on PCA, three principal components explained 93.5% of the cumulative variance. No associations were found between heavy metals or welding fumes and TGCT. In this study, paternal occupational exposure to heavy metals or welding fumes was not associated with TGCT development in their sons.

Abstract Study question What are the motivations, experience, and acceptability of thermal male contraception (TMC) in men using TMC as the only couple contraceptive ? Summary answer Men wished to share the role of providing contraception in a couple and to avoid the use of hormones. Sexual satisfaction was significantly improved. What is known already Two male contraceptive methods inducing spermatogenesis suppression have achieved contraceptive efficacy, on a large population for hormonal contraception and a smaller one for thermal contraception (TC). The suppression of sperm counts to below 1 million/mL results in fertility rates commensurate with female pills. TC is obtained by wearing 15 hours/day a contraceptive underwear (CU) inducing a 2°C increase in testicular temperature (Mieusset &amp; Bujan 1994). As no commercially available male thermal contraceptive exists, little information regarding the acceptability of this approach to men has been reported and only one publication analysed the hypothetical acceptability of TMC (Amouroux et al. 2018). Study design, size, duration We contacted 72 men who had started the CU testing phase between June 2011 and September 2019. The participants completed an anonymous online survey of 93 questions exploring population characteristics, contraceptive histories, motivations for choosing TC, patient experience at the successive phases of use, relations with partner and environment, and satisfaction with TC. Sexual life was evaluated by rating (1 to 4): frequency and pleasure of sexual intercourse, sexual desire, morning and intercourse erections. Participants/materials, setting, methods Among men who had reached the contraception use phase, the quality ratings of sexual life were compared over time, from the contraceptive method used before TC (“N-1” method) to the 2 previous methods (“N-2” and “N-3”)): “N-3” versus “contraception use phase”, “N-3” versus “CU testing phase,” and “CU testing phase” versus “contraception use phase.” These comparisons were made for all 5 sexual life parameters by a paired samples Wilcoxon signed rank test. Main results and the role of chance Sixty-seven men responded (93% response rate);63 participated in the survey (94% participation rate). The median age was 31 [21-52], 30 men (48%) had a Master’s degree or more, 49 (78%) had no child and 24 (38%) no parental project. Among the men whose couple's contraception method immediately before TC (N-1) was a female method (n = 36/63, 57%), the 2 main reasons for switching to a male method were the wish to share the role of providing contraception in a couple (n = 31/36, 86%) and the willingness to take responsibility for contraception (n = 21/36, 58%). The main motivations for choosing TC were “not using hormones” (n = 59/63, 94%) and using a method regarded as “natural” (n = 49/63, 78%). Most of men (68%) adopted the method in less than 2 weeks. During the contraception use phase (n = 59 men), they described: i) significantly improved sexual satisfaction (3,49±0,29/4) compared to that experienced with previous contraceptive methods (3,27±0,58/4;p&amp;lt;0,01); ii) high female partner confidence (100% rather/totally confident partner) and support (88% rather/totally supporting partner); iii) improved self-esteem (n = 35/59, 59%). The main negative point was discomfort, reported by 24% of the men. The overall satisfaction score of using TC was 3.78±0.46/4, and 100% would recommend TC to other men. Limitations, reasons for caution The study population may seem small (63 men). However, it is not different from that of the previously pubished studies on men using hormonal contraception, which included 20, 38, 79 and 57 men, respectively. Our study also lacked a similar questionnaire for the female partners. Wider implications of the findings Men using TC successfully assumed the responsibility for and performed a daily task to provide couple contraception. This positive evaluation of TC together with its previously published contraceptive efficacy and reversibility on a small population, should encourage the development of this method. Trial registration number IRB No. 2020-09-10-005