Initial Semen Parameters Research Articles

Role of seminal MDA, ROS, and antioxidants in cryopreservation and their kinetics under the influence of ejaculatory abstinence in bovine semen

Despite many cryopreservation techniques in bovine semen, various stressors' detrimental effects remain a significant issue. The present study targeted to assess the role of semen quality parameters, sperm function tests, lipid peroxidation, reactive oxygen species (ROS), and different antioxidants in the cryopreservation of bovine semen. Further, the kinetics of lipid peroxidation, ROS, and antioxidants on repeated semen collection under short ejaculatory abstinence were studied. We designed a comparative study where bulls were grouped into good and low freezable semen groups (Freeze-groups) based on their post-thaw motility. All the bulls included had similar initial motility and qualified minimum standards for initial semen parameters viz. semen volume and sperm concentration. The present study detected a higher lipid peroxidation and ROS viz. superoxide anions (•O2-) and a lower total antioxidant capacity (TAC) in the low freeze-group compared to the good freeze-group. The ROS and antioxidants showed unique kinetics on repeated semen collection at short intervals, and no significant change was detected in semen volume, sperm motility, and sperm concentration. This study detected higher head abnormalities and poor acrosome integrity in the low freeze-groups. The present study results indicated that the sperm head might be the most vulnerable part of the sperm to cryopreservation stress. The present study finds significantly higher lipid peroxidation and ROS levels and reduced antioxidant capacity as the primary reasons for low cryopreservability. Further, repeated semen collection with a shorter or lack of abstinence does not impose any significant change in the semen volume and sperm concentration; moreover, it could be beneficial for higher antioxidant levels and lower lipid peroxidation levels. As seminal plasma has both inhibitory and stimulatory roles in sperm function and cryopreservation, identifying the critical role players of seminal plasma and identifying sperm related changes in cryopreservation could predict the cryopreservability potential of semen.

Варикоцелэктомия: критический анализ предикторов восстановления репродуктивной функции

Varicocele is one of the most common correctable cause of male subfertility. According to recent meta-analyzes of RCTs varicocelectomy, in general, leads to significant improvement in semen quality and thereby male fertility. However, varicocelectomy success rate is 55-70%, it means that 30-40% of infertile men does not experience fertility improvement after varicocelectomy. The aim of our review was to search and analyze literature data on clinical-anamnestic and laboratory-instrumental predictors that positively or negatively affect varicocelectomy efficacy. As a result of analysis, we systematized the studied predictors based on the total points calculated on the basis of number and quality of studies that confirm or reject the studied predictor as a significant, into three levels of evidence: predictors of high, moderate and low evidence levels. Predictors of high level of evidence included: initial semen parameters, sperm DNA fragmentation index, and Doppler ultrasound imaging of testicular vessels. The initial serum level of FSH and testicular volume were included in the group of predictors of moderate evidence level. At the same time, such factors as the male age and his female partner age, varicocele grade and infertility duration entered the group of predictors of low evidence level. In this way, we performed gradation of predictors of varicocelectomy efficacy in subfertile men based on the analysis of the evidence level of each predictor.

98 Efficacy of commercial equine semen freezing extenders for cryopreservation of southern white rhinoceros (Ceratotherium simum simum) sperm

Once nearly extinct in the wild, the southern white rhinoceros is currently listed as near threatened by IUCN. This status is likely to change as poaching continues to escalate. To preserve the species’ current genetic diversity, cryopreserving and biobanking white rhinoceros sperm is imperative. The horse is the closest domestic relative of the rhinoceros and a useful model for the development of assisted reproductive technologies, including semen cryopreservation. Two equine semen cryopreservation protocols were compared to a common rhinoceros freezing method. Semen was collected from a single male on 3 occasions by electroejaculation. Initial semen parameters were 86% motility; speed 3.2 (scale 1-5); 89% plasma membrane integrity; and 95% intact acrosomes. Semen was extended 1:1 in INRA 96 (IMV Technologies, L’Aigle, France) before centrifugation at 400×g for 10min. Supernatant was removed and the sperm pellet was subjected to 1 of 2 treatments: resuspension in 500µL of either BotuCrio (Botupharma, Botucatu, Brazil) or Cryomax (ARS Inc., Chino, CA, USA), both containing a proprietary combination of glycerol and an amide as cryoprotectants. Following a 40-min cool at 4°C, extended semen was frozen in vials at a cooling rate of 30°C/min for 3min before LN submersion. Control semen was extended 1:1 in TEST-Y buffer without cryoprotectant and cooled for 2.5h before adding glycerol to a final concentration of 4%. Extended sperm (500µL) was frozen in vials at 12.5°C/min for 15min before LN submersion. Initial motility score (IMS;% motile×speed of progression2), plasma membrane integrity (IPL), and acrosome integrity (IAC) were recorded after extension. All vials were thawed at 37°C for 60s and the cryoprotectant was removed by centrifugation. Sperm pellets were resupended in M199+HEPES and sperm was evaluated for the characteristics described above at 37°C at 0, 30, and 60min (T0, T30, T60) post-thaw. All data are expressed as a percentage of initial (%IMS,%IPL, and%IAC) to account for the differences in sperm parameters between ejaculates. Cryopreservation protocol significantly affected%IMS at T0 (P=0.0131, Table 1). Although the differences were significant only at T0, sperm frozen in Botucrio or Cryomax tended to maintain a higher%IMS than the control freeze at all time points. However, sperm frozen in Cryomax lost a greater percentage of%IMS over time (67% from T0 to T60v. 44 and 46% for Botucrio and TEST-Y, respectively). Cryopreservation protocol did not affect%IAC or%IPL at any time point, but again Cryomax and Botucrio tended to be higher than TEST-Y. This study indicates that rhinoceros sperm may suffer less cryodamage in Botucrio or Cryomax frozen at 30°C/min than in the conventional TEST-Y frozen at 12.5°C/min. Table 1.Percent of initial motility score (IMS), plasma membrane integrity (IPL), and acrosome integrity (IAC) at 0, 30, and 60min post-thaw (T0, T30, and T60, respectively)

Quantitative semen parameters of Atlantic cod (Gadus morhua) and their physiological relationships with sperm activity and morphology

Summary Aspects of sperm motility and metabolism are important for understanding gamete quality. The objectives of this study were to investigate changes in adenosine 5¢-triphosphate (ATP), and lactate dehydrogenase (LDH) of Atlantic cod sperm over time following activation, and to determine if initial semen parameters (ATP, LDH, Na + ,K + ,C a 2+ , total protein, osmolality, pH, and spermatocrit) are indicative of sperm quality. Analyses of ATP and LDH were also performed on sperm cells at 0.5, 2, and 90 min post-activation, while sperm activity was recorded at 0.5 and 2 min. Initial semen parameters did not appear to be strong indicators of sperm quality but some correlations were observed between pH, LDH, K + , and protein and motility parameters in the first two min post-activation. The lack of significant correlation may be attributed to the low variability of almost all parameters observed between males in this study. There was a significant effect of post-activation time on sperm ATP and LDH with both ATP and LDH decreasing with time after activation. The decrease in ATP levels during the course of sperm motility suggests that the metabolic activity is too low to substitute the energy consumed. Continued research on sperm motility and metabolism will lead to a better understanding of fertilization and gamete quality in this species.

Sperm recovering efficiency, a mathematical model, is designed to objectively evaluate semen processing techniques and methods

Objective To propose the sperm recovering efficiency (SRE) model, an equation that can be used to objectively quantify the technical skill of individual operators processing semen specimens in an andrology laboratory. Additionally, this equation can be used to compare different semen processing methods and protocols. This model integrates the semen parameters of initial sperm concentration, motility, morphology, and final sperm recovery to produce a unique number that can be used to compare results. Design A theoretical and deductive analysis. Setting Private assisted reproductive units. Patient(s) None. Intervention(s) None. Main outcome measure(s) Motile progressive and morphologically normal sperm concentration. Result(s) The SRE is a mathematical equation that can be used to compare initial semen parameters to postpreparation concentration, progressive motility, and normal morphology. The result is a number between 0 and 1. The model constitutes an objective means for determining the efficiency of a particular technique (gradients, migration, or filtration) or to set up the best conditions for that technique (time, gravities, medium volume, and sample volume). Additionally, the SRE permits comparisons between different operators, media, lots, laboratory conditions, treatments, and sample parameters. It can be used as a tool in laboratory quality control and for multicenter studies. Conclusion(s) Sperm recovering efficiency (SRE) is an objective method to compare the results of semen processing techniques.

R-178. Heavy insemination IVF and ICSI on sibling oocytes for semen with doubtful fertilizing ability at the first trial: analysis of initial semen parameters

R-178. Heavy insemination IVF and ICSI on sibling oocytes for semen with doubtful fertilizing ability at the first trial: analysis of initial semen parameters

Variation in recovery of motile sperm after preparation by a simple Percoll gradient technique.

Numerous techniques have been used to prepare sperm for assisted reproduction technology. Density-gradient centrifugation with Percoll is becoming a method of choice. This study reviewed the results of a simple two-layer discontinuous Percoll gradient. We reviewed retrospectively results obtained from 208 semen specimens prepared for in vitro fertilization by a discontinuous Percoll gradient. Overall results (mean +/- SD) were a recovery of 21 +/- 16% of total sperm, recovery of 38 +/- 30% of motile sperm, and a motility in the final sperm preparation of 88 +/- 10%. Specimens with higher initial concentrations had higher motility in the final preparation. Higher total recovery and higher motility in the final preparation were found for specimens with a higher initial motility. This simple two-layer Percoll technique is rapid and inexpensive and reliably produces a final sperm preparation with desirable characteristics. Even though specimens with poor initial parameters yielded final preparations with excellent characteristics, fertilization rates were still related to the initial semen parameters.

In Vitro Kallikrein Treatment of Human Spermatozoa: Stimulation of Sperm Penetration into Zona-Free Hamster Eggs Depends on the Quality of Initial Semen Parameters but not on Constituents of the Kallikrein-Kinin System in Seminal Plasma

We investigated the effect of an in vitro kallikrein treatment of human spermatozoa on their ability to penetrate zona-free hamster eggs. Sixty-four infertility patients were classified into four groups (normo-, oligo-, astheno-, and oligoasthenozoospermia) in order to find out which men would benefit most from this procedure. An aliquot of each ejaculate was diluted with Ham’s F-10 medium supplemented with human serum albumin and kept at 22C for 1 hour. Another aliquot was incubated with kallikrein (5KU/ml.) for one hour at 22C. These sperm suspensions were then washed twice, subjected to a swim-up preparation, and subsequently incubated at 37C for 2 hours. The mean penetration rates of kallikrein-treated samples were significantly higher than those of the corresponding control assays in all but the oligoasthenozoospermia group. Our results suggest a direct proteolytic action of kallikrein because the stimulation of penetration into hamster eggs was not correlated with the amount of constituents of the kallikrein-kinin system (kinins, kininogen, kininase) in seminal plasma.

Sperm motion parameters after suppression of spermatogenesis with a gonadotropin-releasing hormone antagonist plus testosterone supplementation

To evaluate whether the chronic administration of a GnRH antagonist supplemented with T enanthate affects sperm motion parameters.Prospective study.Academic medical research environment.Six normospermic men of reproductive age.A 20-week treatment included the administration of 10 mg of GnRH antagonist every day and 25 mg of T enanthate once a week.Computerized sperm motion analysis on each participant every 2 weeks for a period of 11 months.Sperm concentration decreased after 4 weeks of the GnRH antagonist administration, accompanied by a reduction of sperm motility from an initial mean value of 56% to 34% on treatment week 4 and 21% on treatment week 6. In contrast, sperm velocity, linearity, amplitude of the lateral head displacement, and beat cross-frequency values did not significantly change in spite of reduced overall sperm concentration and motility. Azoospermia was reached within 6 to 12 weeks of the GnRH administration, and it was sustained during the treatment period. During recovery of spermatogenesis, sperm movement parameters returned to initial values earlier than sperm concentration and normal morphology. All participants recovered initial semen parameters 20 weeks after the end of treatment.The administration of a GnRH antagonist supplemented with T enanthate successfully induced reversible suppression of spermatogenesis. The sperm motion characteristics appeared to be the last parameters to decline and the first to return to initial values after the treatment.

Sperm motion parameters after suppression of spermatogenesis with a gonadotropin-releasing hormone antagonist plus testosterone supplementation

To evaluate whether the chronic administration of a GnRH antagonist supplemented with T enanthate affects sperm motion parameters. Prospective study. Academic medical research environment. Six normospermic men of reproductive age. A 20-week treatment included the administration of 10 mg of GnRH antagonist every day and 25 mg of T enanthate once a week. Computerized sperm motion analysis on each participant every 2 weeks for a period of 11 months. Sperm concentration decreased after 4 weeks of the GnRH antagonist administration, accompanied by a reduction of sperm motility from an initial mean value of 56% to 34% on treatment week 4 and 21% on treatment week 6. In contrast, sperm velocity, linearity, amplitude of the lateral head displacement, and beat cross-frequency values did not significantly change in spite of reduced overall sperm concentration and motility. Azoospermia was reached within 6 to 12 weeks of the GnRH administration, and it was sustained during the treatment period. During recovery of spermatogenesis, sperm movement parameters returned to initial values earlier than sperm concentration and normal morphology. All participants recovered initial semen parameters 20 weeks after the end of treatment. The administration of a GnRH antagonist supplemented with T enanthate successfully induced reversible suppression of spermatogenesis. The sperm motion characteristics appeared to be the last parameters to decline and the first to return to initial values after the treatment.

Subzonal sperm microinjection in cases of severe male factor infertility and repeated in vitro fertilization failure

To examine (1) fertilization rates obtained with subzonal sperm microinjection when different numbers of sperm are injected into the perivitelline space; (2) when subzonal sperm microinjection is combined with dilute insemination; and (3) the association of semen quality characteristics with fertilization. Subzonal sperm microinjection and subzonal sperm microinjection combined with dilute insemination was performed in 109 and 41 cycles on patients in two clinical trials in Melbourne, Australia, and Bologna, Italy, respectively. PATIENT PARTICIPANTS: Couples who have experienced repeated in vitro fertilization failure or in whom the husband has severe male factor infertility. The number of oocytes fertilized after injection of different numbers of sperm into the perivitelline space, the number of patients transferred, and pregnancy outcome. The injection of multiple numbers of sperm into the perivitelline space failed to improve monospermic fertilization rates but caused an increase in polyspermic fertilization. In patients with initial semen parameters exhibiting greater than 50% motility or greater than 50% normal morphology fertilization rates were improved when subzonal sperm microinjection-treated eggs were incubated in a dilute insemination medium. Six pregnancies were obtained, two of which have progressed to term. When applied to male factor patients, the subzonal sperm microinjection technique results in a 14% to 15% fertilization rate. However, of the 102 embryos transferred only three (2.9%) fetal heart beats were obtained.