P–130 The effect of time of ejaculation and time of processing of semen on fertilization and ongoing pregnancy in IVF/ICSI treatments

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
A Badal ◽  
G Pilgram ◽  
D Diaz de Pool ◽  
L Van der Westerlaken
Keyword(s):  
Incubation Time ◽  
Room Temperature ◽  
Semen Quality ◽  
Sperm Quality ◽  
Gradient Centrifugation ◽  
Handling Time ◽  
Fertility Treatment ◽  
Ongoing Pregnancy ◽  
Ongoing Pregnancy Rate ◽  
Oocyte Vitrification

Abstract Study question Does time between ejaculation and processing, and time between processing and insemination/injection affect fertilization rate (FR) and ongoing pregnancy rate (OPR) in IVF/ICSI treatments? Summary answer Increasing time between processing and insemination significantly decreased the OPR after IVF. FRs after IVF/ICSI and OPR after ICSI were not affected by different time-intervals. What is known already The choice for IVF or ICSI depends on semen quality, however, this doesn’t affect the outcome of IVF/ICSI treatments (Mariappen et al 2018). After ejaculation, the percentage of motile spermatozoa decreases progressively at a rate of about 10%/hour (Makler 1979). According to the ESHRE-guideline, semen should be processed within 1 hour after ejaculation. In our laboratory, a validation was performed that confirmed a decrease in sperm motility after ejaculation. During incubation at 37 °C after processing, the sample remained stable in incubation medium (unpublished data). Therefore, we analyzed the effect of handling time and incubation time with regard to IVF/ICSI outcomes. Study design, size, duration This retrospective data analysis examines the effect of time between ejaculation and processing using density-gradient centrifugation (handling time) and time between processing and insemination (IVF)/injection (ICSI) (incubation time) on the FR and OPR, irrespective of the initial semen quality. A total of 1488 oocyte pickups (844 IVF, 644 ICSI) were included from 1060 patients undergoing fertility treatment between 2017 and 2019. Oocyte pickups without oocytes, with oocyte vitrification, or with donor oocytes were excluded. Participants/materials, setting, methods Anonymized data were obtained from the laboratory database ProMISe. Handling time and incubation time of the semen incubated at 37 °C and 5% CO2 were analyzed in relation to the occurrence of TFF (Total Fertilization Failure), FR and OPR. Linear and logistic regression was performed in SPSS version 25. In case of significant association, the data were adjusted for potential confounders, such as woman’s age, semen quality before and after preparation, and number of oocytes. Main results and the role of chance This study shows that increasing the incubation time of the semen significantly reduced the OPR per ET in IVF treatments (from 30,8% within 3,5 hours to 24,1% after 6 hours) even after adjusting for the potential confouders. However, the OPR in ICSI treatments was not significantly affected by the incubation time (rather, there was an opposite trend). Also, the handling time of the semen did not significant effect the FR per OPU and the OPR per ET in IVF/ICSI treatments. The overall percentage of TFF was 3,5% and did not differ significantly between the IVF and ICSI treatments. Both handling time and incubation time did not have a significant effect on the occurrence of TFF. An explanation for the decrease in OPR in IVF treatments may be that increasing the incubation time at 37 °C reduces the sperm quality as the capacitation reaction takes place too early, energy levels are reduced, DNA damage increases, or vacuoles arise in the sperm heads (Thijssen et al 2014, Jackson et al 2010, Peer et al 2007). Incubation at room temperature and reduction of the insemination time may improve OPR. Limitations, reasons for caution Retrospective study limitations (bias), no data on DNA fragmentation, incubation of semen only at 37 °C after preparation. Wider implications of the findings: Although it is recommended to produce semen at the IVF-department, our results show that an exception can be made, when production of a semen sample in a clinical setting is stressful, with no negative effect on the outcome. Furthermore, incubation-time at room temperature may have a positive effect on OPR. Trial registration number Not applicable

scholarly journals A Modified Flotation Density Gradient Centrifugation Technique Improves the Semen Quality of Stallions with a High DNA Fragmentation Index

Animals ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1973
Author(s):  
Muhammad Umair ◽  
Heiko Henning ◽  
Tom A. E. Stout ◽  
Anthony Claes
Keyword(s):  
Dna Fragmentation ◽  
Semen Quality ◽  
Sperm Quality ◽  
Gradient Centrifugation ◽  
Quality Parameters ◽  
Sperm Dna Fragmentation ◽  
Fragmentation Index ◽  
Sperm Dna ◽  
Cushion Layer ◽  
Dna Fragmentation Index

Sperm DNA fragmentation compromises fertilization and early embryo development. Since spermatozoa lack the machinery to repair DNA damage, to improve the likelihood of establishing a healthy pregnancy, it is preferable to process ejaculates of stallions with a high sperm DNA fragmentation index (DFI) before artificial insemination or intracytoplasmic sperm injection. The aim of this study was to examine a modified flotation density gradient centrifugation (DGC) technique in which semen was diluted with a colloid solution (Opti-prepTM) to increase its density prior to layering between colloid layers of lower and higher density. The optimal Opti-prepTM solution (20–60%) for use as the bottom/cushion layer was first determined, followed by a comparison between a modified sedimentation DGC and the modified flotation DGC technique, using different Opti-prepTM solutions (20%, 25% and 30%) as the top layer. Finally, the most efficient DGC technique was selected to process ejaculates from Friesian stallions (n = 3) with high sperm DFI (>20%). The optimal Opti-prepTM solution for the cushion layer was 40%. The modified sedimentation technique resulted in two different sperm populations, whereas the modified flotation technique yielded three populations. Among the variants tested, the modified flotation DGC using 20% Opti-prepTM as the top layer yielded the best results; the average sperm recovery was 57%; the DFI decreased significantly (from 12% to 4%) and the other sperm quality parameters, including progressive and total motility, percentages of spermatozoa with normal morphology and viable spermatozoa with an intact acrosome, all increased (p < 0.05). In Friesian stallions with high sperm DFI, the modified flotation DGC markedly decreased the DFI (from 31% to 5%) and significantly improved the other semen quality parameters, although sperm recovery was low (approximately 20%). In conclusion, stallion sperm DFI and other sperm quality parameters can be markedly improved using a modified flotation DGC technique employing a 40% Opti-prepTM cushion and a 20% top layer.

scholarly journals The Effect of Incubation Temperature On Semen Parameters Before Intra-Uterine Insemination

2021 ◽  
Author(s):  
Lingling Salang ◽  
Kanok Seejorn ◽  
Thanida Pongsritasana
Keyword(s):  
Body Temperature ◽  
Sperm Motility ◽  
Incubation Time ◽  
Room Temperature ◽  
Incubation Temperature ◽  
Intrauterine Insemination ◽  
Gradient Centrifugation ◽  
Semen Parameters ◽  
Centrifugation Technique ◽  
Optimal Incubation

Abstract Objectives To evaluate the effect of different incubation temperature between room (26–28˚c) and body (37 ˚c) temperature on percentage of progressive sperm motility and the optimal incubation period before intrauterine insemination. Methods Seventy-one normal semen samples under WHO 2010 criteria were recruited. All semen was prepared with Density Gradient Centrifugation technique (DGC) and divided into two groups to evaluate sperm motility at 60 min of incubation time. First group: prepared semen was incubated in room temperature (26˚-28˚c) and second group: prepared semen was incubated in body temperature (37°c). Moreover, each group is divided into 4 items for compare sperm motility between both groups in same of incubation time and evaluate the optimal incubation time between the items in the same groups. Results Spermatozoa incubated at body temperature had a significantly higher percentage of progressive sperm motility than those incubated at room temperature (89.62 ± 8.02 vs 85.97 ± 9.42; p < 0.01). The optimal incubation time at room temperature was 30 minutes and at body temperature was 60 minutes. These results suggest that spermatozoa incubated at 37°C for 60 minutes were more likely to have better sperm motility functions for IUI. Conclusion These results suggest that spermatozoa incubated at 37°C for 60 minutes were more likely to be effective for use in IUI in terms of sperm motility functions.

scholarly journals The Effects of In Vitro Incubation of Asthenoteratozoospermic Semen after Density Gradient Centrifugation at Room Temperature and 37C on Sperm Parameters, Chromatin Quality and DNA Fragmentation in a Short Time Period

2020 ◽  
Author(s):  
Motahareh Karimi Zarchi ◽  
Behnam Maleki ◽  
Mahmood Dehghani Ashkezari ◽  
Leila Motamed Zadeh ◽  
Azam Agha-Rahimi
Keyword(s):  
Dna Fragmentation ◽  
Density Gradient ◽  
Room Temperature ◽  
Density Gradient Centrifugation ◽  
Sperm Quality ◽  
Gradient Centrifugation ◽  
Sperm Parameters ◽  
Sperm Dna Fragmentation ◽  
In Vitro Incubation

Background: Sperm quality is an important factor in assisted reproductive technology (ART) that affects the success rate of infertile couples treatment. In vitro incubation of sperm can influence its parameters and DNA integrity. The present study focused on the effect of different incubation temperatures sperm parameters on asthenoteratozoospermia semen prepared with density gradient centrifugation at different times. Methods: Twenty-seven samples were collected and prepared. Then, the suspension was divided into two parts. One part was incubated at room temperature (RT), and another was incubated at 37°C. Immediately and after 2 hr (2H) and 4 hr (4H), sper-matozoa were evaluated regarding motility, viability, morphology, sperm protamine deficiency, chromatin and DNA fragmentation. Statistical analysis was performed using paired t-test and repeated measures. The p<0.05 was considered statistically significant. Results: Our results showed that following 2 and 4 hr of incubation at RT, sperm progressive motility and viability decreased significantly. Sperm DNA fragmentation increased significantly following 2 and 4 hr of incubation at RT and 37°C. The Trend analysis confirmed that there were no significant differences between sperm parameters and DNA fragmentation after different times at RT and 37°C. Conclusion: Incubation of sperm at RT in comparison to 37°C didn’t preserve sperm parameters and DNA efficiently. Therefore, IVF, ICSI and IUI procedure should be performed in the soonest possible time after sperm preparation.

The Prevalence of Bacteriospermia in Infertile Men and Association with Semen Quality in Southwestern Iran

2020 ◽  
Vol 20 (2) ◽  
pp. 198-202 ◽  
Author(s):  
Mohammad Motamedifar ◽  
Yalda Malekzadegan ◽  
Parisa Namdari ◽  
Behzad Dehghani ◽  
Bahia Namavar Jahromi ◽  
...  
Keyword(s):  
Semen Quality ◽  
Sperm Quality ◽  
Reproductive Function ◽  
World Health ◽  
Public Health Issue ◽  
Control Group ◽  
Microbiological Analysis ◽  
Male Reproductive Function ◽  
Infertile Men ◽  
Southwestern Iran

Introduction: Infertility considered as a social and public health issue and estimated that most of these infertile couples are residents of developing countries. Infectious diseases including the history of Sexually Transmitted Infections (STIs) may impact on male reproductive function. Therefore, the present study aimed to investigate the prevalence of bacterial contaminants of semen and probable association with sperm quality of infertile men in Iranian population. Methods: The study population consisted of 200 infertile men and 150 fertile men attending an infertility Center in southwestern Iran during the study period in 2015. The assessment of sperm parameters was according to the World Health Organization (WHO) guidelines. The presumptive pathogens were identified using standard microbiology tests and confirmed by specific PCR primers. Results: The prevalence of bacteriospermia in the semen of the infertile group was significantly higher than that in the fertile group (48% vs. 26.7%, P <0.001). The microbiological analysis of samples showed that the most abundant species of bacteria in semen of infertile men were Chlamydia trachomatis (12.5%) followed by Neisseria gonorrhoeae (11%). On the other hand, in the control group, Lactobacillus spp. (17.3%) was the most isolated pathogen. Results showed that the presence of N. gonorrhoeae, C. trachomatis, Mycoplasma genitalium, Haemophilus, and Klebsiella was significantly associated with sperm abnormality. Conclusion: Based on our findings, it seems that bacteriospermia is associated with alterations in the properties of semen which may lead to a decrease in the fertilization potential of sperm. Therefore, immediate and appropriate treatment is necessary before investigating every other possible cause of infertility.

Restraint stress of male mice triggers apoptosis in spermatozoa and spermatogenic cells via activating the TNF-α system

Zygote ◽  
2020 ◽  
Vol 28 (2) ◽  
pp. 160-169 ◽  
Author(s):  
Jie Zhang ◽  
De-Ling Kong ◽  
Bin Xiao ◽  
Hong-Jie Yuan ◽  
Qiao-Qiao Kong ◽  
...  
Keyword(s):  
Psychological Stress ◽  
Restraint Stress ◽  
Semen Quality ◽  
Sperm Quality ◽  
Fertilization Rate ◽  
Seminiferous Tubules ◽  
Spermatogenic Cells ◽  
Male Mice ◽  
Testicular Cells ◽  
Tnf Α

SummaryStudies have indicated that psychological stress impairs human fertility and that various stressors can induce apoptosis of testicular cells. However, the mechanisms by which psychological stress on males reduces semen quality and stressors induce apoptosis in testicular cells are largely unclear. Using a psychological (restraint) stress mouse model, we tested whether male psychological stress triggers apoptosis of spermatozoa and spermatogenic cells through activating tumour necrosis factor (TNF)-α signalling. Wild-type or TNF-α−/− male mice were restrained for 48 h before examination for apoptosis and expression of TNF-α and TNF receptor 1 (TNFR1) in spermatozoa, epididymis, seminiferous tubules and spermatogenic cells. The results showed that male restraint significantly decreased fertilization rate and mitochondrial membrane potential, while increasing levels of malondialdehyde, active caspase-3, TNF-α and TNFR1 in spermatozoa. Male restraint also increased apoptosis and expression of TNF-α and TNFR1 in caudae epididymides, seminiferous tubules and spermatogenic cells. Sperm quality was also significantly impaired when spermatozoa were recovered 35 days after male restraint. The restraint-induced damage to spermatozoa, epididymis and seminiferous tubules was significantly ameliorated in TNF-α−/− mice. Furthermore, incubation with soluble TNF-α significantly reduced sperm motility and fertilizing potential. Taken together, the results demonstrated that male psychological stress induces apoptosis in spermatozoa and spermatogenic cells through activating the TNF-α system and that the stress-induced apoptosis in spermatogenic cells can be translated into impaired quality in future spermatozoa.

scholarly journals Macro- and microelements in serum and seminal plasma as biomarkers for bull sperm cryotolerance

2021 ◽  
Vol 63 (1) ◽  
Author(s):  
Maja Zakošek Pipan ◽  
Petra Zrimšek ◽  
Breda Jakovac Strajn ◽  
Katarina Pavšič Vrtač ◽  
Tanja Knific ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Semen Quality ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Quality Characteristics ◽  
Freezing And Thawing ◽  
Additional Tool ◽  
Bull Semen ◽  
Bull Sperm ◽  
Fresh Semen

ABSTRACT Background Wide variation in fertility rates is observed when using frozen bull semen, even when the bulls have met quality standards for semen production. Therefore, a simple and reliable test to assess the freezing potential of bull semen based on the analysis of fresh semen or blood would be of great value. Attention is now turning to assessment of seminal plasma components such as proteins and elements. In the present study, the concentrations of macro- and microelements in fresh bull semen plasma and in serum and their correlation with quality characteristics of fresh semen and with semen quality after freezing and thawing were determined. Ejaculates were collected from 30 mature bulls, and semen volume, concentration, sperm motility, morphology, tail membrane integrity, plasma membrane permeability and DNA fragmentation were determined on the day of collection and after freezing and thawing. The concentrations of macroelements (Na, Mg, K and Ca) and microelements (Cu, Fe, Zn and Se) were determined in the seminal plasma and serum. The semen samples were classified into satisfactory and unsatisfactory groups according to the fresh semen quality. Results Zinc and Se levels measured in serum were associated with almost all fresh and frozen-thawed semen quality characteristics, while Fe levels were associated only with acrosomal defects in fresh semen. Zinc and Fe levels in fresh seminal plasma were associated with various quality characteristics of fresh and frozen-thawed semen, while Se level in fresh seminal plasma was not associated with any of the semen quality characteristics. Conclusions Microelements were shown to be useful as biomarkers involved in the analysis of bull sperm quality and could be used as an additional tool to predict bull semen quality after freezing and thawing. Our results confirm that the analysis of Zn and Se levels in serum and Zn, Cu and Fe levels in fresh seminal plasma can provide information to discriminate between bull semen samples with spermatozoa with high or low cryotolerance.

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