Swim-up Research Articles

Effect of semen washing methods on diminishing the transmission of viral infections in artificial reproductive technology

Seminal infections were considered to be an etiologic factor of male infertility. Semen washing is used as an appropriate technique in artificial reproductive technology (ART) for serodiscordant couples when the partner of male is contaminated. The aim of this study was to investigate the role of semen washing methods in reducing the transmission of viral infections in ART. In this narrative review we investigated accessible information from Google Scholar, PubMed, Scopus, Web of Science, IranMedex, Irandoc, Magiran, SID, MEDLIB and Scientific Information Database until 2019. The MeSH terms of semen washing, HIV, assisted reproduction, serodiscordant, and virus transmission were used. According to the search strategy, 89 articles were discovered. After checking the titles, abstracts and manuscripts, a collection of 45 papers were chosen pursuant to the suitability indexes. The studies were performed around different categories of semen washing (swim up, density gradient) in serodiscordant couples. Some studies highlighted that semen washings can completely prevent viral transmission, but a few do not believe this claim. However, scientific researchers have revealed that sperm washing may diminish the possibility of infection in serodiscordant couples where the male partner is infected. Swim up in combination with density gradient is a suitable method to diminish the transmission of infections.

EFFECT OF SWIM-UP AND GLASS WOOL TECHNIQUES, WITH ADDING ANTIOXIDANTS TO TRIS EXTENDER ON IMPROVING POST-CRYOPRESERVED TOTAL SPERM CHARACTERISTICS IN STRAW AND FREEZABILITY PERCENTAGE FOR LOW SEMEN QUALITY OF HOLSTEIN BULLS

This study was conducted to investigate the effect of swim-up and glass wool as sperm separation techniques with adding vitamin E and superoxide dismutase (SOD) to Tris extender on improving post-cryopreserved total sperm characteristics in straw and freezability percentage for low semen quality of Holstein bulls. Low and good of semen were extended using Tris extender. Good semen quality (GSQ) was divided into 3 groups (L1; Tris extender, L2; 2 mM vitamin E, L3; 200 IU SOD) Low semen quality (LSQ) was divided into two main groups, and subdivided into 3 sub-groups (L4; Tris extender, L5; 2 mM vitamin E, L6; 200 IU SOD). In the second main group, swim-up and glass wool techniques were used with adding vitamin E and SOD and subdivided into 3 sub-groups with each technique, and referred to L7, L8 and L9 for swim-up technique and L10, L11 and L12 for glass wool technique. Sperms with good motile intact acrosome and plasma membrane normal morphology of sperm were obtained using the glass wool separation technique with adding vitamin E.

Sperm Quality and Deoxyribonucleic Acid Fragmentation after 5 and 10 min Centrifugation with Swim-Up Processing Technique: A Prospective Cohort Study

AIM: This study aimed to assess the difference in sperm concentration, total motility, progressive motility, and deoxyribonucleic acid (DNA) fragmentation index (DFI) before and after processing with 5 and 10 min swim-up (SU). METHODS: Fifty patients who met the study inclusion criteria from June 2020 to October 2020 were subjected to routine semen and sperm DNA fragmentation analysis. Each of the samples was then divided into three tubes, one as control and the others were processed using the SU method with 5 and 10 min centrifugation time, respectively. After being processed, the samples were subjected again to routine semen and sperm DNA fragmentation analysis. The results were being compared among three groups. RESULTS: The sperm concentration after 5 and 10 min SU (27.78–39.79 and 35.36–51.09, respectively; p < 0.05) was significantly higher compared to fresh semen (24.85–32.33). The total motility before and after 5 and 10 min SU were 43.78–51.08, 97.66–98.20, and 97.86–98.20, respectively. The progressive motility after 5 and 10 min SU (0–41 and 0–54, respectively) was significantly higher than fresh semen (0–24; p < 0.05). The DFI was significantly better after 5 min SU (3.82–6.98) compared to fresh semen and after 10 min SU (13.48–19.04 and 1–25, respectively; p < 0.05). CONCLUSION: Prolonged centrifugation time may yield a higher number of sperm concentration and motility, but it may also lead to a higher DFI. Hence, a shorter centrifugation time should be used for a better semen quality intended for assisted reproductive technologies.

P–013 Sperm freezing does not affect live birth rates: results from 6,594 cycles in normozoospermic patients

Abstract Study question Does sperm cryopreservation influence the reproductive outcomes of normozoospermic patients undergoing elective ICSI? Summary answer After controlling for confounders, the use of cryopreserved semen from normozoospermic patients does not affect pregnancy and live birth rates after ICSI. What is known already Sperm cryopreservation with slow freezing is a common practice in ART. While frozen-thawed semen typically presents reduced motility and vitality, its use for ICSI is generally considered adequate in terms of reproductive outcomes. Nevertheless, most studies comparing reproductive outcomes between fresh versus cryopreserved sperm include patients with oligo- and/or asthenozoospermia, where the altered quality of the sample can partially mask the full effect of freezing/thawing. The objective of this study is to ascertain whether ICSI using fresh or cryopreserved semen from normozoospermic patients results in similar fertilization rates and reproductive outcomes. Study design, size, duration Retrospective cohort of 6,594 couples undergoing their first elective ICSI cycle between January 2011 and December 2019, using normozoospermic partner semen (fresh or cryopreserved). All cycles involved a fresh embryo transfer, either at cleavage or blastocyst stage. Cycles were divided in 4 groups: fresh semen with partner’s oocytes (FSPO, n = 1.878), cryopreserved semen with partner’s oocytes (CSPO, n = 142), fresh semen with donor oocytes (FSDO, n = 2.413), and cryopreserved semen with donor oocytes (CSDO, n = 2.161). Participants/materials, setting, methods A slow freezing protocol using GM501 SpermStore medium (Gynemed, Lensahn) was used for all sperm cryopreservation. Sperm washing, capacitation, and selection prior to ICSI were performed equally for fresh and frozen-thawed samples, using pellet swim-up in IVF® medium (Vitrolife, Göteborg). Fertilization rate (FR), pregnancy (biochemical, clinical, and ongoing) and live birth (LB) rates were compared among study groups using Pearson’s Chi square and Student’s t-test. A p-value <0.05 was considered statistically significant. Main results and the role of chance Male and female age, sperm concentration and motility after ejaculation, and number of oocytes inseminated were similar between study groups compared (FSPO vs. CSPO, FSDO vs. CSDO). As expected, oocyte donation cycles resulted in higher LB rate than cycles in which partner’s oocytes were used (30.04% vs 18.17%, p < 0.001). In cycles using partner’s oocytes, no significant differences were observed between fresh and cryopreserved sperm in FR, pregnancy and LB rates (p > 0.05 for all outcomes). However, in oocyte donation, the mean FR after ICSI using cryopreserved semen (73.6 ± 19.6) was lower than the FR obtained with fresh semen (75.1 ± 19.2), p = 0.010. Similarly, in oocyte donation cycles, the biochemical pregnancy rate was significantly lower when using cryopreserved semen (48.5% in CSDO vs. 52.3% in FSDO, p = 0.009), while clinical, ongoing pregnancy and LB rates were similar between both semen status (p > 0.05). In oocyte donation, a subgroup analysis including only the ICSI cycles with embryo transfer at blastocyst stage (n = 1.187 for FSDO, n = 337 for CSDO) confirmed that the LB rate was comparable between fresh and cryopreserved semen groups (34.7% vs 35.6% respectively, p = 0.76), without significant differences in pregnancy rates neither (p > 0.05 for all outcomes). Limitations, reasons for caution Caution should be exerted when extrapolating these results to different protocols for sperm cryopreservation and selection, or to IVM and classical IVF cycles, which were excluded from analysis. Due to the retrospective nature of the study, some uncontrolled for variables may affect the results. Wider implications of the findings: Sperm cryopreservation does not affect pregnancy and live birth rates in normozoospermic patients, although it may lower slightly fertilization rates. In line with previous studies including patients with an apparent male factor detected after routine semen analysis, sperm cryopreservation is a safe and convenient technique. Trial registration number Not applicable

P–012 A new sperm preparation technique for ICSI

Abstract Study question Is there any difference in this technique(swim-in) according to clinical results in ICSI cycles? Summary answer Fertilization rates are significantly better with this new technique(swim-in). There is no difference in pregnancy and ongoing pregnancy rates between swim-in and DGS techniques. What is known already Sperm separation is crucial in assisted reproductive technologies, based on different principles like migration, filtration, or density gradient centrifugation techniques. However, there are some studies that centrifugation steps and using gradient solutions may increase sperm DNA fragmentation. On the other hand, multiple contacts to different plastic surfaces such as pipettes, plastic tubes, etc., and also pipetting the ejaculate in sperm preparation may have detrimental effects. The ideal sperm separation technique should be non-invasive, not time-consuming, easy, and cost-effective, not cause sperm damage or non-physiological alterations of the separated sperm cells, eliminate dead spermatozoa and other cells, including leukocytes. Study design, size, duration This is a prospective randomized study between 01/02/2019...01/01/2021. Sperms were prepared either with this new technique that we call “swim-in”(n = 359) or with density gradient centrifugation(n = 404) before microinjection (ICSI). Fertilization rates, clinical pregnancy rates, and on-going pregnancy rates were compared between groups. Sperm motility is less than%20 patients are not included in the study. t-test and Chi-square test used. p < 0.05 significant. Participants/materials, setting, methods For this new centrifugation free technique(swim-in), a Braun injector, 21G green needle, and sperm washing medium are used. First, we pulled a 0.5 ml sperm wash medium in a Braun injector. After sperm liquefaction, we put Braun injector with the needle in the sperm cap perpendicularly and waited 30 minutes at 37-degree Celcius. Sperms swam through the needle to the injector used for ICSI. For DGC sperms were prepared according to the 2015 WHO manual. Main results and the role of chance: We try to make a centrifugation-free, gentle, cost-effective, and easy sperm preparation technique and called it a swim-in technique. Groups were comparable according to mean age, collected oocyte numbers, and oocyte maturation ratios. Fertilization rates are%73,2(1786/2438) vs%66,9(2167/3237) respectively for swim-in and DGC groups. Clinical pregnancy rates are%45,1 vs%42,3 respectively for swim-in and DGC groups. On-going pregnancy rates are%41,5 vs%39,6 for swim-in and DGC respectively. There is a significant difference between the two techniques according to fertilization rates but no difference according to clinical pregnancy and ongoing pregnancy rates. Limitations, reasons for caution Sperms can move by their own motility from ejaculate to the sperm washing medium for that reason with severe sperm problem patients are not suitable for this technique. Wider implications of the findings: This technique is centrifugation free so there are no detrimental g-forces. The needle used may mimic the physiological environment like Uterine Tube. Alternative sperm washing mediums like follicular fluid or chemical attractor progesterone rich medium can be used.DNA fragmentation rates and live birth rates should be established for future aspect. Trial registration number Not applicable

Model-based localization of deep-diving cetaceans using towed line array acoustic data.

Passive acoustic monitoring using a towed line array of hydrophones is a standard method for localizing cetaceans during line-transect cetacean abundance surveys. Perpendicular distances estimated between localized whales and the trackline are essential for abundance estimation using acoustic data. Uncertainties in the acoustic data from hydrophone movement, sound propagation effects, errors in the time of arrival differences, and whale depth are not accounted for by most two-dimensional localization methods. Consequently, location and distance estimates for deep-diving cetaceans may be biased, creating uncertainty in abundance estimates. Here, a model-based localization approach is applied to towed line array acoustic data that incorporates sound propagation effects, accounts for sources of error, and localizes in three dimensions. The whale's true distance, ship trajectory, and whale movement greatly affected localization results in simulations. The localization method was applied to real acoustic data from two separate sperm whales, resulting in three-dimensional distance and depth estimates with position bounds for each whale. By incorporating sources of error, this three-dimensional model-based approach provides a method to address and integrate the inherent uncertainties in towed array acoustic data for more robust localization.

Effect of Microfluidic Sperm Separation vs. Standard Sperm Washing Processes on Laboratory Outcomes and Clinical Pregnancy Rates in an Unselected Patient Population

A prospective, multicenter, randomized, sibling oocyte study was conducted with 86 couples to evaluate if a microfluidic sperm separation device improved ICSI sperm selection and subsequent cycle outcomes of fertilization, blastocyst utilization, ploidy, and clinical pregnancy rate when applied to a general patient population. Patients with at least 10 metaphase II oocytes were enrolled in the study and sibling oocyte groups were split in half. One half of the oocytes underwent ICSI with the control processed sperm and the other half were injected with sperm sorted by the ZyMōt microfluidic sperm separation device. Fertilization rate was recorded and resulting blastocysts were biopsied and evaluated for ploidy status with NGS. Euploid, non-mosaic embryos were randomly selected for single embryo transfer. A total of 787 oocytes were evaluated in the ZyMōt group and 777 in the control group. No statistical differences were observed between ZyMōt and control processing methods in any of the study outcomes evaluated. It is possible that the selection of normal, progressive sperm for ICSI, and the repair capacity of oocytes are sufficient to promote normal embryonic development in the general infertility population.

Improving Sperm Oxidative Stress and Embryo Quality in Advanced Paternal Age Using Idebenone In Vitro-A Proof-of-Concept Study.

Advanced paternal age is associated with increased sperm reactive oxygen species (ROS) and decreased fertilization and pregnancy rates. Sperm washing during infertility treatment provides an opportunity to reduce high sperm ROS concentrations associated with advanced paternal age through the addition of idebenone. Sperm from men aged >40 years and older CBAF1 mice (12–18 months), were treated with 5 µM and 50 µM of idebenone and intracellular and superoxide ROS concentrations assessed. Following in vitro fertilization (IVF), embryo development, blastocyst differentiation, DNA damage and cryosurvival, pregnancy and implantation rates and fetal and placental weights were assessed. Five µM of idebenone given to aged human and mouse sperm reduced superoxide concentrations ~20% (p < 0.05), while both 5 and 50 µM reduced sperm intracellular ROS concentrations in mice ~30% (p < 0.05). Following IVF, 5 µM of idebenone to aged sperm increased fertilization rates (65% vs. 60%, p < 0.05), blastocyst total, trophectoderm and inner cell mass cell numbers (73 vs. 66, 53 vs. 47 and 27 vs. 24, respectively, p < 0.01). Treatment with idebenone also increased blastocyst cryosurvival rates (96% vs. 78%, p < 0.01) and implantation rates following embryo transfer (35% vs. 18%, p < 0.01). Placental weights were smaller (107 mg vs. 138 mg, p < 0.05), resulting in a larger fetal to placental weight ratio (8.3 vs. 6.3, p = 0.07) after sperm idebenone treatment. Increased sperm ROS concentrations associated with advanced paternal age are reduced with the addition of idebenone in vitro, and are associated with improved fertilization rates, embryo quality and implantation rates after IVF.

EFFECT OF SWIM-UP AND GLASS WOOL TECHNIQUES, WITH ADDING ANTIOXIDANTS TO TRIS EXTENDER ON IMPROVING POST-CRYOPRESERVED SOME SEMEN ATTRIBUTES OF LOW SEMEN QUALITY FOR HOLSTEIN BULLS

This study was conducted to investigate the effect of swim-up and glass wool as sperm separation techniques with adding vitamin E and superoxide dismutase (SOD) to Tris extender on improving some post-cryopreserved semen quality for Holstein bulls. Low and good of semen were extended using Tris extender. Good semen quality (GSQ) was divided into 3 groups (L1; Tris extender, L2; 2 mM vitamin E, L3; 200 IU SOD) Low semen quality (LSQ) was divided into two main groups, and subdivided into 3 sub-groups (L4; Tris extender, L5; 2 mM vitamin E, L6; 200 IU SOD). In the second main group, swim-up and glass wool techniques were used with adding vitamin E and SOD and subdivided into 3 sub-groups with each technique, and referred to L7, L8 and L9 for swim-up technique and L10, L11 and L12 for glass wool technique. Improving sperms’ cell motility and live sperm and reducing total sperms’ abnormalities percentages of low semen quality were noticed using swim-up and glass wool techniques were used with adding vitamin E and SOD. In conclusion, glass wool filtration separates out dead, abnormal and immotile sperm cells, a good sperm harvested by this technique.

IGF-I and NGFβ enhance in vitro progressive motility and vitality of human spermatozoa.

PurposeProgressive motility (PM) and vitality are positively associated with fertilization ability of spermatozoa. Here, the effects of IGF‐I and NGFβ on PM and vitality of human spermatozoa were investigated.MethodsForty‐three volunteers gave semen samples after 2‐3 days of sexual abstinence. Each sample was processed with density gradient centrifugation and sperm washing. The pellet was divided into 3 aliquots. An aliquot containing one million of progressively motile spermatozoa was incubated for an hour (37°C) in standard culture medium (control group), and two aliquots with the same number of progressively motile spermatozoa were incubated in medium supplemented with IGF‐I or NGFβ. Two concentrations of IGF‐I (100 ng/ml and 1000 ng/ml) and NGFβ (0,5 ng/ml and 5 ng/ml) were tested.ResultsBoth growth factors significantly increased PM and vitality in comparison with control either at the low or the high concentration. IGF‐I seemed to be more effective than NGFβ. The effects did not seem to be dose dependent with the exception of the effect of IGF‐I on vitality.ConclusionsThe enhancement of PM and vitality of human spermatozoa by IGF‐I and NGFβ opens new ways for the improvement of sperm processing. Further research is needed to determine the most effective concentrations.

Cryopreservation and Preparation of Thawed Spermatozoa from Rhesus Macaques (Macaca mulatta) for In Vitro Fertilization

Advances in assisted reproductive technologies in rhesus macaques have allowed the development of valuable models of human disease, particularly when combined with recent techniques for gene editing. While the ability to perform in vitro fertilization (IVF) in rhesus macaques is well established, this procedure has not yet been optimized. Specifically, damage to the sperm caused by cryopreservation (cryodamage) may lead to unsuccessful artificial insemination and low fertilization and blastocyst formation rates in vitro. To address this, we systematically assessed 2 cryopreservation methods and 4 recovery methods in the following 3 interdependent experiments: 1) comparing sperm survival after vitrification or slow-freezing; 2) comparing simple wash (SW), density gradient centrifugation (DGC), swim-up (SU), and glass wool filtration (GWF) for removal of cryoprotectants and isolation of motile sperm after thawing; and 3) evaluating the efficacy for IVF of the 2 best methods of isolating thawed sperm. We found that after vitrification, only 1.2 ± 0.3% of thawed sperm were motile, whereas after slow-freezing, 42 ± 5% of thawed sperm were motile. SW was significantly better than all other isolation methods for the recovery of total sperm and for the recovery of sperm with an intact plasma membrane. The isolation methods had no significant differences in the recovery of motile sperm or sperm with progressive motility. However, IVF of ova with sperm recovered by DGC resulted in 5% more embryos and 25% more blastocysts than did IVF with sperm recovered by SW. Although additional studies are required to optimize sperm cryopreservation in rhesus macaques, our study showed that slow-freezing, coupled with DGC, provided the highest efficacy in providing functional sperm for in vitro use.

A microfluidic approach to rapid sperm recovery from heterogeneous cell suspensions

The isolation of sperm cells from background cell populations and debris is an essential step in all assisted reproductive technologies. Conventional techniques for sperm recovery from testicular sperm extractions stagnate at the sample processing stage, where it can take several hours to identify viable sperm from a background of collateral cells such as white bloods cells (WBCs), red blood cells (RBCs), epithelial cells (ECs) and in some cases cancer cells. Manual identification of sperm from contaminating cells and debris is a tedious and time-consuming operation that can be suitably addressed through inertial microfluidics. Microfluidics has proven an effective technology for high-quality sperm selection based on motility. However, motility-based selection methods cannot cater for viable, non-motile sperm often present in testicular or epididymal sperm extractions and aspirations. This study demonstrates the use of a 3D printed inertial microfluidic device for the separation of sperm cells from a mixed suspension of WBCs, RBCs, ECs, and leukemic cancer cells. This technology presents a 36-fold time improvement for the recovery of sperm cells (> 96%) by separating sperm, RBCS, WBCs, ECs and cancer cells into tight bands in less than 5 min. Furthermore, microfluidic processing of sperm has no impact on sperm parameters; vitality, motility, morphology, or DNA fragmentation of sperm. Applying inertial microfluidics for non-motile sperm recovery can greatly improve the current processing procedure of testicular sperm extractions, simplifying the fertility outcomes for severe forms of male infertility that warrant the surgery.

The evaluation of Human papilloma virus and human herpes viruses (EBV, CMV, VZV HSV-1 and HSV-2) in semen samples.

There are a number of risk factors, especially viral diseases, which can lead to infertility. Among the various viral infections, much attention has been given to the role of the Papillomaviridae and Herpesviridae. After collecting 82 semen samples (37 teratospermia, 2 asthenozoospermia, 2 oligoasthenospermia, 1 oligospermia, 6 asthenoteratospermia and 34 normal semen samples), and washing them, the DNA from both freshly ejaculated spermatozoon and washed spermatozoa was extracted. Subsequently, the prevalence of EBV, CMV, HSV-1, HSV-2, VZV and HPV was evaluated using Multiplex PCR and Nested PCR. In this study, 1 normal and 5 abnormal semen samples were infected with HSV-1 (1 normal, 4 teratospermia and 1 oligoasthenospermia). In addition, there were 2 VZV-positive samples (both were teratozoospermia). Nested PCR indicated that 1 asthenozoospermia, 1 asthenoteratospermia, 3 teratospermia and 4 normal samples were HPV positive (including 8 HPV-18 and 1 HPV-33). Among 9 HPV-positive subjects, 3 samples were negative after washing the infected samples. The prevalence of EBV, CMV, VZV, HSV-1 and HSV-2 remained unchanged prior to and after washing. Maybe sperm washing can be useful to eliminate HPV infection from semen samples, but further investigation is required because of the small number of samples.

Female reproductive system and oogenesis in the mite Bakericheyla chanayi

The fine structure of the female reproductive system of a cheyletid mite Bakericheyla chanayi (Trombidiformes: Cheyletidae) is investigated for the first time. This system consists of an unpaired ovary, glandular oviduct, receptaculum seminis, long cuticle-lined vagina, and genital atrium terminating in the genital opening. A separate sperm access system has not been found. The receptaculum seminis opens into the distal oviduct region, where fertilization apparently takes place. The ovary contains clusters of oogonia (cystocytes), clustered early meiotic cells, a few growing previtellogenic oocytes, and 3 large nurse cells. The dorsal ovarian region is occupied by the clusters of bacteriocytes which harbor symbiotic bacteria. Oocytes undergo vitellogenesis in individual ovarian pouches, each connected to the corresponding nurse cell by an intercellular bridge. The fine structure of the bridge suggests transport between the interconnected cells in the course of vitellogenesis. The population of cystocytes was shown to be heterogenic. The electron-light cells enter meiosis and develop into the oocytes or nurse cells. The electron-dense cystocytes do not show meiotic transformation and probably give rise to the bacteriocytes. The early development of the nurse cells and oocytes is similar and accompanied by the blebbing of the nuclear envelope, appearance of nuage material and Balbiani bodies.

Sperm Parameters, ASAs and Apoptosis After Processing by the Double Tube and Swim up Methods.

The aim of this study was to improve the quality of semen samples by using a novel double-tube (DT) method. The DT method was developed to select sperm and compared with traditional swim-up (SU) technique for 31 semen samples. Sperm DNA integrity were tested with TUNEL and SCSA. Content of antisperm antibodies (ASA) in the semen was measured by ELISA and MAR. Levels of the caspase-3 in the sperm were assessed by western blotting. After SU and DT, 15 couples and 16 couples were underwent IVF-ET. The number of RCDs, the percentage of SDF and DFI, ASA and the level of caspase-3 were significantly decreased after DT and SU (p = .001 and p < .001). When the DT and SU compared, there were significant changes in the number of RCD, the percentage of SDF and DFI, ASA and the level of caspase-3 (p < 0.05-0.001). There was a higher cleavage rate (p = .017) and a lower abortion rate (p < .05) in DT-IVF group than in SU-IVF group. DT selection yielded spermatozoa with low RCDs, DFI, ASA, and caspase-3 which would be benefit for ART.

Assessing the impact of COVID 19 on male fertility: A systematic review

Coronavirus disease 2019 (COVID-19) infection is presently a global pandemic; it is mainly an acute respiratory infection. It can affect various organs of the body, primarily the lung, GI tract, and CVS and can even affect the urinary system and male reproductive system. A systematic search using PubMed, EMBASE, and Scopus was performed using the keywords and Boolean operators. There were 55 studies, which were found to be relevant to the search conducted in the above-mentioned databases. Out of the 55 studies, 32 were identified to be fitting the inclusion and exclusion criteria of the review. Recent studies have conflicting results on the presence of COVID-19 in the semen of patients who were reverse transcription polymerase chain reaction (RT-PCR) positive for COVID-19. Majority studies negate COVID-19 infection in semen as per RT-PCR, but semen parameters are adversely affected especially in moderate cases. One study states that RT-PCR is positive in few semen samples of severe cases. Testes have high angiotensin-converting enzyme 2 (ACE2) expression and transmembrane serine protease 2 is essential for priming of the spike (S) protein in COVID-19 virus and it is concentrated in spermatogonia and spermatids, which means if spermatogonia is infected with COVID-19 and it hampers the spermatogenesis, it can lead to dysfunction of the male reproductive system. There is evidence of high ACE2 expression in renal tubules, which means there can be renal tubular damage because of COVID-19 infection and there is a possibility of shedding of virus in urine. The COVID-19 detected in the semen could also be because of shedding of virus from the urinary tract in men. The authors thus conclude that there is a need larger study to answer the questions like whether the COVID-19 infection just present in the semen or it is attached to the spermatozoa or its present inside the cells because of interaction of COVID-19 S protein and ACE2 receptor on the spermatogonia. If COVID-19 infection is detected in the semen, then whether it can be removed from the semen by the sperm washing techniques and is it safe to use in ART procedures or any other fertility treatment.

Low density Porcicoll separates spermatozoa from bacteria and retains sperm quality

Antibiotics are added to semen extenders to control the growth of bacteria contaminating semen during collection but may contribute to the development of antibiotic resistance. An alternative would be physical separation of spermatozoa from bacteria. The objective of the present study was to evaluate two low densities of Porcicoll for removal of bacteria, and for their effect on sperm recovery and sperm quality. Semen was collected from boars at a commercial station. Aliquots of 8 extended ejaculates were subjected to colloid centrifugation through 20% Porcicoll (P20) and 30% Porcicoll (P30) in 500 mL tubes and then stored at 17 °C. Microbiological examination and sperm quality evaluation (computer assisted sperm analysis and flow cytometry) were carried out on controls and all colloid-selected samples immediately after preparation and again after storage for 3 and 7 days. The microorganisms found were mainly bacteria from the environment, gut or skin. There was a considerable reduction or complete removal of some bacteria by both colloids. Recovery rates were 86% for P20 and 81% for P30. Sperm quality was not adversely affected by colloid centrifugation on day 0, and thereafter showed a more gradual deterioration in colloid centrifuged samples than in controls, possibly due to lower bacterial contamination. There were no differences in sperm quality between the two colloid treatments. Thus, these results show that contaminating bacteria in semen can be controlled by centrifugation through low density colloids.

Comparison of three sperm selection methods for ICSI-DGC, Cumulus column, and incubation with supernatant product of adipose tissue-derived adult stem cells: An experimental study.

BackgroundThe examination of sperm parameters and sperm DNA integrity are necessary for male fertility expression. These parameters can be affected by method of sperm separation.ObjectiveTo measure the damage of each sperm separation method on the sperm parameters and sperm DNA integrity.Materials and MethodsIn this experimental study, semen samples of 20 infertile men with asthenoteratozoospermia (Infertility Research Center, Qom, Iran, 2017) were processed in three ways: density gradient centrifugation (DGC), cumulus column, and incubation with supernatant products of adipose tissue-derived adult stem cells (SPAS). The results of sperm parameters and DNA fragmentation before and after the process were statistically analyzed.ResultsThe number of separated sperms by normal morphologies during the SPAS and the cumulus column was significantly more than the corresponding population in the DGC group. In addition, although all three methods have the same ability to increase total sperm motility and the number of recovered sperms, in the field of forwarding movement and DNA fragmentation, the SPAS method performed more efficiently (p = 0.021).ConclusionSperm parameters and DNA fragmentation in the SPAS group were better than those in the DGC and cumulus column groups. Furthermore, it has been shown that the sperm capacity was increased with the SPAS method. However, the rearrangement of sperm chromatin by reducing the disulfide bridges and providing the possibility of re-histone over capacity causes a significant reduction in DNA fragmentation.

Epigenetic Status of Lebanese Dizygotic Twins

Lebanon is an eastern Mediterranean country with a small population of only 4 million (Abbas et al. 2020). It is characterized for its distinct human genetic, cultural, ethnic, and religious diversity (Haber et al. 2011). The average live twinning births rate in Lebanon is 18.9/1000 (Romanos &amp; Borjac, 2018). Twins can be either monozygotic (MZ) or dizygotic (DZ). The difference between them is how they are formed during embryogenesis (Hoekstra et al. 2008). When a single sperm fertilizes a single egg, it splits then in two genetically identical twins (Xu et al. 2015). However, in fraternal twins case, separate eggs are fertilized by separate sperm to yield genetically different twins (Martino et al. 2013).

Use of Androcoll-ETM to Separate Frozen-Thawed Llama Sperm From Seminal Plasma and Diluent.

It is not easy to separate frozen-thawed South American camelid sperm from seminal plasma (SP) and diluents to be used for in vitro embryo production. The objective of this study was to evaluate Androcoll-E™ (AE) efficiency to separate llama sperm from SP and freezing extender in frozen-thawed semen. A total of 22 ejaculates from five Lama glama males were collected using electroejaculation. After performing semen analysis (sperm motility, concentration, viability, membrane function, and acrosome integrity), samples were cryopreserved with a diluent containing lactose, ethylenediaminetetraacetic acid (EDTA), egg yolk, and 7% dimethylformamide. After thawing, samples were divided in aliquots, one of which was used as a control and the others processed by AE. Experiment 1 (12 ejaculates): 100 μl of frozen-thawed semen was placed on top of 1,000 μl AE column and centrifuged at 800 g for 10 min. Experiment 2 (10 ejaculates): two samples of 100 μl of frozen-thawed semen were placed on two columns of 500 μl AE each, and both were centrifuged at 800 g for 10 and 20 min, respectively. Pellets were resuspended in Tyrode's albumin lactate pyruvate (TALP) medium, and sperm parameters were evaluated. A significant decrease in all sperm parameters was observed in thawed samples compared to raw semen. AE allowed the separation of frozen-thawed sperm from SP and freezing extender independently from the height of the column used and time of centrifugation assayed. Although no significant differences were found between AE columns, higher sperm recovery was observed with 500 μl of AE coupled with 20 min of centrifugation. Despite the significant decrease observed in sperm motility in AE samples, no changes in sperm viability, membrane function, and acrosome integrity were observed when comparing control thawed semen with the sperm recovered after AE (p > 0.05). The use of AE columns, either 500 or 1,000 μl, allows the separation of frozen-thawed llama sperm from SP and freezing extender, preserving the viability, membrane function, and acrosome integrity. Of the protocols studied, 800 g centrifugation during 20 min using a 500 μl column of AE would be the method of choice to process frozen-thawed llama semen destined for reproductive biotechnologies.