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.

Freezability of Goat Epididymal Sperm using Aloe vera Extract and Trehalose in Diluents

Background: Cryopreservation process causes oxidative stress on sperm membranes, which in turn damages sperm organs and enzymatic activities which thereby decrease motility, functional membrane integrity and sperm fertility. Therefore, current study carried out to evaluate the effect of Aloe vera Ethanolic Extract (AEE), alone and with trehalose in diluents on cryopreserved epididymal goat sperm. Methodology: Epididymal sperm isolated from testes with motility >70% and total morphological abnormalities <10%. The experimental treatments consist of control (no additives) and basic diluents plus 5, 10, 20 or 50 μg/ml of AEE (AEE1, AEE2, AEE3 and AEE4, respectively), tr (150 mM trehalose), tr+AEE1, tr+AEE2, tr+AEE3 and tr+AEE4. Results: Obtained data show that the extender containing AEE3, AEE1+tr, AEE2+tr and AEE3+tr improved significantly the cryopreserved sperm. The combined treatments indicate also a decrease in MDA than control. In addition, AEE2+tr and AEE3+tr showed the lowest (P<0.05) DNA fragmentation compared to the other treatments. Extender containing AEE3+tr resulted in higher total motility and viability than the extender containing tr alone, as well as AEE1, AEE2 and AEE4 treatments. Conclusion: The present study indicates that ethanolic extract of Aloe vera could be used for goat sperm cryopreservation. Also, it can be concluded that trehalose in combination with 20 μg/ml of Aloe vera extract can be promised cryoprotectant in goat epididymal sperm freezing.

Use of glycosaminoglycans from Oreochromis niloticus skin as an antioxidant supplement for milt cryopreservation of Brazilian bocachico

The study aimed to evaluate the in vitro antioxidant action of glycosaminoglycans (GAGs) from the skin of Oreochromis niloticus, and to determine their ideal concentration to supplement the sperm freezing medium of Prochilodus brevis. In experiment 1, the in vitro antioxidant properties of GAGs were verified through the analysis of DPPH, chelating ferrous ability, and total antioxidant capacity. In experiment 2, milt pools were formed, which were frozen in solution supplemented or not with different GAGs concentrations: 0 (control), 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, or 5.0 mg mL-1 (total of 10 treatments). The samples were evaluated for membrane integrity, DNA integrity, sperm morphology, and sperm kinetics. The results of experiment 1 showed that the GAGs exhibited, with the increase of the concentration, significant antioxidant action, for all the evaluated tests, mainly in the chelating ferrous ability. In experiment 2, it was observed that the increase of GAGs concentration decreased kinetic parameters (P < 0.05), however, the control and 0.5 mg mL-1 GAGs concentration showed similar results. For the other parameters (membrane integrity, DNA integrity, and sperm morphology), there was no decrease in results with the increase of GAGs concentration. In conclusion, GAGs extracted from O. niloticus skin have antioxidant action, and the concentration of 0.5 mg mL-1 was the most adequate to supplement the P. brevis sperm-freezing medium.

P–126 A comparison of the efficacy of sperm freezing using high security tubes versus high security straws

Study question Are there any advantages in using High security tubes rather than High Security straws for conventional slow sperm freezing? Summary answer Freezing sperm in High Security tubes (HST) improved post-thaw recovery rate and motility, and also reduced processing and handling compared to High Security straws (HSS). What is known already The use of High Security freezing consumables (HSFC) in an IVF setting is a safe and effective way of eliminating concerns related to viral cross-contamination during storage. The lower diameter of HSS does make them susceptible to warming during handling. The HSFC used in this study is the only CE marked products that are made of resin, leak-proof and shatter-proof in all cryogenic temperatures even in LN2. No previous studies have compared the use of HST with HSS for conventional human sperm freezing. This study sets out to investigate the performance of HST compared to HSS. Study design, size, duration The study was designed as a controlled split-sample study with blind post-thaw analysis. Following the routine WHO analysis of 20 semen samples, the remainder of each of the samples was evenly divided and cryopreserved by conventional slow freezing in each of the two different HSFC. The freeze was conducted simultaneously by the same practitioner, employing the same freezing protocol and cryoprotectant. The pre-freeze and post-thaw concentration, total and progressive sperm motility were recorded. Participants/materials, setting, methods At one IVF clinic, semen samples with sperm density ≥15million/ml, ≥40% motility, ≥1.5ml were included. Cryoprotectant (SpermFreeze, Fertipro) was added dropwise to unprepared semen and kept at room temperature for 10 minutes before loading into HSFC (0.5ml CBS™HSS; CBS™HST). HSFC were heat-sealed (SYMS; SYMSIII sealers) and placed in vapour for 30 minutes before plunging into LN2. Samples were thawed by immersion in a 37Cº water bath for 5 minutes and analysed using WHO methods. Main results and the role of chance Paired-t test was used to compare the percentage motility between the different HSFC. All analysis was considered statistically significant when p &lt; 0.01. We demonstrated that the sperm recovery rate (Percentage total motility post-thaw/ Percentage total motility pre-freeze) in HST was 66.63 ± 14.94 (mean ± standard deviation) compared to 40.80 ± 14.69 in HSS. In the HSS, the percentage post-thaw total motility was 19.99 ± 7.21 and the percentage post-thaw progressive motility was 12.26 ± 2.59. In the HST, the percentage post-thaw total motility was 32.57 ± 8.33 and the percentage post-thaw progressive motility was 23.08 ± 5.53. The overall improvement when using HST against HSS was 12.53 ± 5.69, 10.44 ± 5.29 for the total motility and the progressive motility respectively. Comments were recorded regarding the handling and the condition of the HSS and HST for each freeze event. Neither device displayed any leakage of LN2 or any explosion during the warming. The freezing process was easier and faster using HST rather than HSS. It was also noted that the entire sample can be recovered from the HST, unlike the HSS. Limitations, reasons for caution The study looked at sperm recovery in terms of motility only. DNA damage was not considered as a parameter of sperm quality. Also, fertilization, pregnancy rates, live birth rates and the use of poorer quality sperm samples have not been investigated. Wider implications of the findings: For conventional sperm freezing, the use of HST resulted in improved sperm motility and progression post-thaw, when compared to HSS. This finding supports the use of HST to improve the post thaw quality of sperm, benefitting patients with own frozen samples, recipients of donor sperm and donor sperm banks. Trial registration number Not applicable.

Rare Sperm Freezing

Gamete cryobanking has been widely incorporated in present assisted reproductive technology (ART). Preserving male gametes for future fertility is considered to be an easy and accessible way to insure one’s reproduction. Despite the fact that the method could not secure success, sperm freezing could be the only chance to father biological offspring. In cases when severe male factor (SMF) infertility is diagnosed (retrograde ejaculation, virtual azoospermia, obstructive azoospermia, cryptozoospermia) and providing fresh semen samples for assisted reproduction may alter chances to achieve pregnancy, rare sperm cryopreservation could contribute for conceiving. Isolation, selection and cryopreservation of single sperm cells from semen samples is a challenging procedure. Different approaches and devices could be used in order to extract utmost spermatozoa. Aiming to highest cryosurvival rates sperm freezing protocols should be carefully considered. For some men, rare sperm cryopreservation might be the only alternative for parenting biological offspring. Thus, the latter technique should be widely discussed, developed and practiced in assisted reproduction.

For Whom Does the Clock Tick?: Male Repro-Temporality in Fertility Campaigns, Scientific Literature, and Commercial Accounts

Sperm swimming in circles or a lone sperm cell with two heads: male reproductive aging is increasingly equated with poor sperm quality, the prevalence of offspring learning disabilities even schizophrenia. To discuss the construction of a male biological clock, this article asks: how does the biological clock intervene in men’s reproductive bodies. And secondly: how is male repro-temporality visually and rhetorically invoked in fertility campaigns, in medical scientific accounts and in the marketing material of one elective sperm-freezing company? Situated within an interdisciplinary theoretical framework, the article draws upon biomedicalization theory (e.g. Clarke et al. 2003), reproductive masculinity studies (e.g. Daniels 2006; Almeling and Waggoner 2013), and social scientific theorizing of time and temporality (e.g. Amir 2006; van de Wiel 2014a; 2014b) to discuss the emergence of male repro-temporality. This article contributes to the interdisciplinary scholarly agenda on time and temporality by theorizing the emergence of a male biological clock as a type of repro-temporality that, in its discursive and aesthetic framing, portrays male reproductive aging as involving loss and disability. The article concludes that while the biological clock derives its temporal force from the logic of decay, it simultaneously cements heteronormative ideals of the nuclear family, re-naturalizes the genetic unit, and situates men as proactive and modern in their anticipation of future infertility.

Optimizing the conventional method of sperm freezing in liquid nitrogen vapour for Wallachian sheep conservation program

The aim of the present study was to optimize the conventional method of sperm freezing in liquid nitrogen (LN₂) vapour for successful cryopreservation of Wallachian ram sperm, the genetic resources of the Czech Republic. Sperm in straws were frozen using the conventional freezing method via a static exposure of sperm doses to LN₂ vapour, or by four different modified freezing methods. Under modified freezing, straws were frozen by a discontinuous, time-dependent decremental change in the distance between the straws and the surface of LN₂. The viability of sperm was evaluated by flow cytometry after sperm equilibration, and immediately after thawing. Besides the observed inter-sire and daily variation, the obtained results suggest the methodological weakness of the conventional freezing method via the static exposure of sperm doses to LN₂ vapour. With the use of the optimized freezing procedure, all parameters of thawed sperm were significantly (P &lt; 0.05) improved in comparison with the conventional method: percentage of thawed sperm viability increased up to 48.3%, percentage of sperm with plasma membrane damage after thawing decreased to 6.58%, percentage of sperm with acrosome damage decreased to 24.4%, and percentage of sperm with deteriorated mitochondrial activity decreased to 6.28%. In conclusion, our results suggest that an optimized freezing procedure should be routinely used instead of the conventional method to cryopreserve Wallachian ram sperm.