Search

This study was conducted to examine the effects of adding glutathione (1 mM) to media used for sperm washing and in vitro fertilization (IVF) on the improvement of early development of embryos produced using cryopreserved spermatozoa of the less IVF-competent bull (the one considered unqualified as spermatozoa supplier for the production of bovine blastocysts using IVF). The cryopreserved spermatozoa of this bull were characterized by normal motility and lower ATP content and blastocyst productivity than those of IVF-competent bulls. The addition of glutathione to the sperm washing medium was more effective in improving the productivity of blastocysts and ATP content than the addition of glutathione to the IVF medium or no glutathione addition at all (control). These results suggest that this simple method may be used to improve the potential of cryopreserved spermatozoa of less IVF-competent bulls to fertilize oocytes in vitro and to induce normal embryonic development after fertilization.

Objective To investigate whether the presented rheotaxis-based microfluidic device could be used to separate spermatozoa from viruses (i.e., Zika) in the infected semen sample during the selection and washing process. Design Quantitative and experimental study of the sperm washing/selection process through the microfluidic platform exploiting the positive rheotaxis of sperm. Setting None. Patient(s) None. Intervention(s) None. Main Outcome Measure(s) Human sperm were purchased from a sperm bank. The raw semen sample was mixed with viruses and loaded into a microfluidic device. Experiments were performed with 2 different flow rates (0 and 25 μL/minute) to investigate the washing efficiency of the device in the sperm selection process. The sperm sample was collected after 45 minutes and analyzed to check whether the collected sample is free of any infections (viruses) after isolation. Result(s) Fluorescent microscopy and quantitative polymerase chain reaction-based analysis showed that the sperm selected with the presented rheotaxis-based microfluidic device at the optimal flow rate (25 μL/minute) was free of any viruses. Conclusion(s) We have developed a simple, cost-effective microfluidic device that mimics the conditions of the female genital tract while washing out the raw semen efficiently during the selection process for assisted reproductive technology.

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.

Purpose Progressive 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. Methods Forty‐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. Results Both 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. Conclusions The 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.

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.

OBJECTIVE: In this study, the effects of sperm parameters on the success of intrauterine insemination were investigated. STUDY DESIGN: The data from 309 infertile couples who were admitted between 2012-2018 without a female factor were analyzed retrospectively and included in the study. After the administration of gonadotropin and hCG (5000-10000 IU), single insemination was performed in 36-40 hours in all cycles. All couples underwent routine infertility screening. The relationship between sperm parameters (motility, morphology, sperm count), patient age, duration of infertility with intrauterine insemination success was evaluated.RESULTS: There was no statistically significant difference between the two groups in terms of mean age and age related-parity. There was no statistically significant difference between male ages, liquefaction, and sperm volumes between the two groups (p=0.898, p=0.448, p=0.651). Before washing; There was a statistically significant difference between the sperm concentration, percentage of total motile sperm, percentage of progressive motility sperm, percentage of normal sperm morphology, and total sperm count between the two groups (p=0.0001, p=0.0001, p=0.0001, p=0.0001, p=0.0001). After sperm washing; the results were similar to those obtained before washing. While statistically significant difference was observed between sperm volume and sperm concentrations (p=0.023, p=0.018), no significant difference was observed between the two groups in total sperm count (p=0.612).CONCLUSION: As a result, during the application of intrauterine insemination to infertile couples, total motile sperm count, progressive motility sperm count ratio and high sperm ratio with normal morphology used in order to increase pregnancy success can be considered as criteria that increase the chances of success.

Research question Is it possible, by sperm-washing spermatozoa from clinically HPV-positive men, to obtain spermatozoa free of human papillomavirus (HPV) to be employed in assisted reproduction? Design This was an observational study performed on HPV-positive men. Freshly ejaculated semen was collected and readily processed by gradient separation followed by swim-up from the washed pellet. The resulting fractions were seminal plasma, cell pellet, round cells, non-motile spermatozoa and motile spermatozoa. All fractions were then tested for the presence of HPV DNA. Results Of the 15 clinically HPV-positive subjects, 67% were positive in at least one of the seminal fractions. If any postivity was detected, the plasma was always HPV positive. No consistent pattern was observed throughout different samples in the cell pellet, round cell and non-motile spermatozoa fractions. However, after the sperm-wash procedure, the fraction of motile spermatozoa was never found to be HPV-positive. Conclusions The sperm-washing technique, which was previously successfully used to remove human immunodeficiency virus, can efficiently remove HPV from spermatozoa. However, the present study was conducted on a small population so a larger follow-up study is recommended. HPV screening should be performed in sperm samples and, upon HPV positivity, sperm-washing should be considered before assisted reproduction techniques are used.

Background: In this study, our aim is to investigate the possible protective effects of curcumin on potential damage caused by benzo(a)pyrene (BaP) on rat sperm motility and morphology.Methods: 36 Wistar Albino male rats were divided into six groups: Group 1, Control; Group 2, Corn oil (BaP dissolver); Group 3, DMSO (Curcumin dissolver); Group 4, BaP; Group 5, Curcumin; Group 6, BaP+Curcumin. By the end of the six week oral administration period, left caudal epididymis and ductus deferens tissues were placed in Pure Sperm Wash (Nidacon) solution (at 37°C) and dissected. Sperms were counted and stained. Measurements of the perinuclear ring of the manchette and the posterior portion of the nucleus were performed. All of the parameters were evaluated statistically.Results: BaP administration caused a decrease in the number of total sperms and total motile sperms, while increasing abnormalities in tail, acrosome and cytoplasmic droplet particularly. Abnormalities did not approach the control group by curcumin administration in expected rates, but the percentage reduced. The number of total and motile sperms increased by curcumin. All parameters were statistically supported.Conclusions: In conclusion, curcumin is one of the antioxidant that could be used potentially to inhibit the effects of Benzo(a)pyrene, but is ineffective in preventing all abnormalities.

Background The restriction of reproductive rights of HIV-positive couples in low-resource settings could be related to the attitudes and skills of health workers. We assessed health workers’ knowledge of safer conception and their attitudes toward the reproductive rights of HIV-positive couples in a tertiary hospital in Nigeria. Methods A cross-section of health workers (n=294) was interviewed using structured questionnaires. Knowledge and attitude scores were analyzed. Logistic regression was employed to generate adjusted odds ratios (AORs) for predictors of attitude. Results Safer conception methods mentioned by respondents included timed unprotected intercourse with (27.9%) and without antiretroviral pre-exposure prophylaxis (37.4%), in vitro fertilization plus intracytoplasmic sperm injection (26.5%), and sperm washing and intrauterine insemination (24.8%). The majority (94.2%) of health workers acknowledged the reproductive rights of HIV-infected persons, although (64.6%) strongly felt that HIV-infected couples should have fewer children. Health workers reported always/nearly always counseling their patients on HIV transmission risks (64.1%) and safer conception (59.2% and 48.3% for females and males, respectively) (p Conclusions Health workers had limited knowledge of safer conception methods, but were supportive of the reproductive rights of HIV-positive couples. Health workers in Nigeria require training to effectively counsel couples on their reproductive rights, risks and options.

Objectives Sperm cryopreservation has great potential for male infertility treatment as used in assisted reproduction technology (ART). There are a variety of cryopreservation methods in order to preserve sperm in a long term. Although conventional freezing and vitrification now are used widely, they have damage on sperm parameters as well as sperm DNA integrity. It is necessary to answer which method is better and appropriate for sperm cryopreservation. The aim of this study was to compare the effect of conventional freezing and vitrification regarding to motility, vitality and morphology of sperm found in washed and unwashed samples. Study design One hundred and five human fresh semen samples were divided into washed and unwashed halves using density-gradient centrifugation. Each group then was split into two aliquots: one cryopreserved by conventional freezing and the other by vitrification, using SpermFreeze Solution™ (Vitrolife, Vastra Frolunda, Sweden) containing glycerol as a cryoprotectant. The sperm parameters were analyzed and compared between six groups: washed fresh (FW), unwashed fresh (FU), washed conventional freezing (CfW), unwashed conventional freezing (CfU), washed vitrification (VitW) and unwashed vitrification (VitU) samples. Results Sperm progressive motility, vitality and normal morphology significantly decreased, together with an appreciable increase in sperm head, midpiece and tail defects when comparing to the fresh sperm parameters after thawing in all groups. In conventional freezing method groups, progressive motility and vitality were substantially higher than that in vitrification method groups. However, vitrification gave better results in normal morphology rates. Additionally, sperm head, midpiece and tail defects were significant lower in two vitrification groups in comparison with conventional freezing groups. Interestingly, washed groups had better sperm parameters than unwashed groups so that washing process before frozen seemed to improve sperm parameters. Conclusion Conventional freezing method resulted in better motility, viability in both washed/unwashed groups. On the contrary, spermatozoa undergoing vitrification were healthier regarding morphology with less defects than conventional freezing. Sperm washing before frozen was a beneficial preparation to sperm cryopreservation.

The aim of this study was to compare the post-thaw distribution of motile sperm subpopulations, following simple or colloid centrifugation. A new analysis was used to evaluate the available number of sperm from each subpopulation after each centrifugation protocol. Frozen/thawed semen samples were divided into the following after-thawing treatments: uncentrifuged control (UDC), sperm washing (SW) and two colloid centrifugation procedures (Equipure, SLC-E, and Androcoll, SLC-A). Percentage of total and progressive motility (TM and PM), as well as sperm motility kinematics, distribution of motile sperm subpopulations, and recovery rates, were statistically compared among treatments. The SLC treatments showed higher (P < 0.001) TM and PM than UDC and SW. Following each SLC procedure, different percentages of the subpopulation with the most vigorous and progressive sperm (sP4) were obtained. SLC-A recovered a larger number of sperm belonging to sP4 than SLC-E, but not significantly higher than SW. From a practical point of view, sperm washing, the standard centrifugation procedure for equine semen processing, recovered the same amount of fast and progressive sperm as colloid centrifugation, apparently the best treatment according to traditional analysis. In conclusion, samples processed by SLC have higher motility percentages than SW and UDC but, after combining the available number of sperm, SLC and SW techniques are equally efficient in recovering sperm from the most vigorous, fast and progressive motile subpopulation (sP4).

To analyze the effect of time intervals from the end of sperm collection to IUI on the pregnancy rates in couples who treated by COH-IUI cycles with gonadotropin due to unexplained infertility.556 couples who underwent the first COH-IUI treatment with gonadotopin due to unexplained infertility were included. Semen samples were obtained by masturbation in a private room near the laboratory on the day of insemination. For each semen samples, time between the delivery time to the laboratory and starting time of sperm washing procedure was noted as semen collection to sperm washing (SC-SW) interval, the time between the starting time of sperm washing procedure and insemination time was noted as sperm washing to insemination (SW-IUI) interval and the time between the delivery time to the laboratory and insemination was noted as semen collection to insemination (SC-IUI) interval. Clinical pregnancy was defined as positive pregnancy test followed by the presence of an intrauterine gestational sac with fetal cardiac activity by transvaginal ultrasonography at least 4 weeks after IUI.Among 556 couples, there were 84 cases of succesful clinical pregnancy and pregnancy rate was calculated as 15.1%. Intervals from SW to IUI and SC to IUI were significantly shorter in pregnant women than in non-pregnant (p0.001 and p=0.007, respectively). SW performed ≤40min after SC resulted in higher clinical pregnancy rates than did SW performed40min after SC (19.1% vs. 6.3%, respectively) (p0.001). IUI performed ≤60min after the end of SW had higher pregnancy rates (16.9%) compared to IUI performed60min after the end of SW (4.0%) (p=0.017). Likewise, IUI performed ≤90min after SC resulted in higher pregnancy rates than did IUI performed90min after SC (18.9% vs. 12.0%, respectively) (p=0.029).IUI outcome is enhanced by shorter intervals from SC to SW, from SW to IUI, and from SC to IUI in gonadotropin-IUI cycles for unexplained infertile couples.

International audience; Retrograde ejaculation (RE) is a condition defined as the backward flow of the semen during ejaculation, and when present can result in male infertility. RE may be partial or complete, resulting in either low seminal volume or complete absence of the ejaculate (dry ejaculate). RE can result from anatomic, neurological or pharmacological conditions. The treatment approaches outlined are determined by the cause. Alkalinizing urinary pH with oral medications or by adding sperm wash media into the bladder prior to ejaculation may preserve the viability of the sperm. This article provides a step-by-step guide to diagnose RE and the optimal techniques to retrieve sperm. Copyright

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.

Objective: The ZyMōt sperm separation device has proven favorable for use in elevated DNA fragmentation index (DFI) male factor patients, as an alternative to density gradient (DG) washing or surgically attained testicular sperm. In 2020, without fully understanding the infectivity and transmission potential of SARS-CoV2 in semen, a more liberal application of a timed ZyMōt microfluidic swim-up was applied to our IVF patients to dilute out and minimize potential pathogens. This study aimed to evaluate whether the use of ZyMōt sperm improved normal embryo development. Materials and Methods: Retrospective analysis of PGT-A/ICSI cycles (N= 3219) between 2016-2020 was conducted to assessed fertilization rates (FR), blastocyst development/utilization rates (BUR) and genetic outcomes. Sperm preparations were performed per standard operating or manufacturer advised (i.e., ZyMōt) procedures. Cumulus oocyte complexes were harvested 36h post-hCG, stripped and ICSI performed 3-5hr later. Zygotes were assessed at 16-18hr post-ICSI, and embryos cultured under humidified tri-gas incubation for up to 7 days. Blastocyst (BL) development as evaluated, and expanded BL or greater were biopsied on Days 5, 6 or 7. All BL were vitrified and genetics determinations for euploidy, aneuploidy and mosaicism were contrast. Applying Chi-squared analysis, we compared potential differences (p

Computer-assisted semen analysis (CASA) is an automated and objective method of evaluating structural (e.g. morphology) and functional sperm parameters (e.g. motility and hyperactivation). Sperm hyperactivation is essential for successful fertilization and is thus an important aspect in determining the fertility potential of a male. In the current study, CASA was used for standard semen analysis and for comparison of the ability of phosphate buffered saline (PBS), BO sperm wash (10 mM caffeine), 4% lignocaine, and 5 mM procaine hydrochloride to induce hyperactivation in Tankwa goat spermatozoa. Twenty-nine ejaculates were collected from randomly selected male goats by electroejaculation. Although none of the four media affected percentage total sperm motility, lignocaine caused a significant decrease (P >0.05) in percentage progressive motility. Exposure to procaine resulted in an increase in swimming speed (P ≤0.05) and star-spin motility tracks, which are typical of sperm hyperactivation. Using PBS and procaine motility data from individually selected spermatozoa, receiver operator characteristic curves were constructed to distinguish the kinematic parameters employed as cut-off values for sperm hyperactivation. PBS and BO sperm wash did not induce hyperactivation (0.1 + 0.2% and 0.04  0.2% respectively), while lignocaine induced little hyperactivation (3.4 + 3.0%) and procaine hydrochloride had the highest percentage hyperactivation (25.3 + 13.6%). The large variation in hyperactivation (0–54.5%) may reflect inter-individual differences in sperm quality among these males. This study indicated procaine hydrochloride was the most promising hyperactivation-inducing medium for Tankwa goat spermatozoa and should be considered for similar assessments in other animal species Keywords: computer-aided sperm analysis, procaine hydrochloride, sperm kinematics, sperm morphometry, sperm motility

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 &gt, 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 &lt, 0.05), while both 5 and 50 µM reduced sperm intracellular ROS concentrations in mice ~30% (p &lt, 0.05). Following IVF, 5 µM of idebenone to aged sperm increased fertilization rates (65% vs. 60%, p &lt, 0.05), blastocyst total, trophectoderm and inner cell mass cell numbers (73 vs. 66, 53 vs. 47 and 27 vs. 24, respectively, p &lt, 0.01). Treatment with idebenone also increased blastocyst cryosurvival rates (96% vs. 78%, p &lt, 0.01) and implantation rates following embryo transfer (35% vs. 18%, p &lt, 0.01). Placental weights were smaller (107 mg vs. 138 mg, p &lt, 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.

Funder: Isaac Newton Trust, Funder: British HIV Association, It is now established that people living with HIV who have an undetectable viral load and adhere to antiretroviral treatment cannot transmit HIV to their sexual partners. Previous research has shown that ‘being undetectable’ changes how HIV‐positive gay men experience their sex lives. But how does it affect gay men’s reproductive behaviours? And what influence does it have on views about parenthood at a time when gay fatherhood has become more socially accepted and publicly visible? Drawing on qualitative interviews with patients and clinicians at four HIV clinics in London, we identify differences in how interviewees talked about the possibility of having children for HIV‐positive men. Both groups, unprompted, frequently referred to sperm washing as a method enabling safe conception. However, whereas clinicians talked about sperm washing as an historical technique, which is no longer necessary, patients spoke of it as a current tool. The men rarely mentioned being undetectable as relevant to parenthood and, when prompted, some said that they did not fully understand the mechanics of HIV transmission. Our findings offer new insights into how biomedical knowledge is incorporated into people’s understandings of living with HIV, raising important questions about how the meanings of being undetectable are communicated.

PROBLEM Artificial insemination, which requires cryopreservation of semen, is not completely optimized in goats because bucks discharge a small volume of ejaculate and seminal plasma (SP) contains specific proteins that are detrimental to spermatozoa at cryopreservation. However, it is not known the effects of sperm washing (removal of SP) before cryopreservation on the post-thawing frozen spermatozoa of Black Bengal bucks. Moreover, it is completely unknown whether SP of goats contains TGF-β and CXCL10 that have been proven essential for fertility in other mammals. METHODS Thirty-five ejaculates were collected from six mature Black Bengal bucks at one-week intervals and were subjected to microscopic evaluation for semen characteristics at pre- and post-freezing condition. The concentrations of TGF-β and CXCL10 in the SP using ELISA were determined. SP was harvested with centrifugation of fresh semen at 1500 g for 15 minutes twice at room temperature. RESULTS Semen characteristics were significantly varied between bucks. Seminal plasma of all ejaculates contained TGF-β and CXCL10 while significant variation of concentrations between bucks was observed in case of CXCL10. Cryopreservation of semen reduced total motility and progressive motility, while sperm washing before cryopreservation was beneficial to the total motility and progressive motility of post-thawing spermatozoa. CONCLUSION Black Bengal buck seminal plasma was affluent of TGF-β and CXCL10 and washing of spermatozoa before cryopreservation was beneficial to the post-thawing sperm motility. The results of the current investigation will be helpful for future research on the roles of SP in female reproductive tract and pregnancy in Black Bengal goats.

A semen analysis evaluates the classical “descriptive” characteristics of ejaculates produced by masturbation. In vivo, the potentially functional sperm population is separated from liquefied semen by virtue of their migration into cervical mucus. To maximize the yield of this method, multiple tubes with relatively small volumes of semen per tube are used, maximizing the total interface area between semen and culture medium. The semen to be used for underlayering is gently aspirated into a sterile plastic syringe of a brand known not to have any deleterious effects upon spermatozoa. The swim-up from semen procedure remains the simplest means of obtaining populations of highly motile human spermatozoa and is that recommended by the World Health Organization’s for sperm fertilizing ability tests. Standardized and quality-controlled methods must be used for semen analysis as for any other laboratory test and there is no substitute for thorough training and experience.

The purpose of this prospective non-randomized study was to study the effectiveness of semen washing followed by intrauterine insemination (IUI) in Human Immune Deficiency Virus (HIV)-discordant couples in which the male partner was infected, in preventing HIV transmission to uninfected partner and offspring. The study was performed in a private assisted reproductive center specialized in couples with infectious diseases and enrolled sixty-nine fertile couples in which male partner tested positive for HIV, seeking for reproductive treatment. Triple sperm washing followed by viral RNA purification and real-time polymerase chain reaction was performed prior to IUI intervention. HIV transmission to female partner and newborns, and clinical pregnancy rate were the main outcome measures. A total of 180 IUI treatment cycles were performed in 69 couples. There were 16 clinical pregnancies (clinical pregnancy rate/cycle 9.0%, clinical pregnancy rate/patient 23.2%), one of which resulted in miscarriage (6.3%). No seroconversion was detected in the 69 women treated with sperm washing followed by IUI or in any of the newborns (tested at birth and at 3 months of age). Sperm washing followed by IUI is a safe and effective treatment option for serodiscordant couples wishing to conceive and to prevent HIV virus transmission to the mothers and newborns.

The use of cooled semen is relatively common in goats. There are a number of advantages of cooled semen doses, including easier handling of artificial insemination (AI) doses, transport, more AI doses per ejaculate, and higher fertility rates in comparison with frozen AI doses. However, cooled semen has a short shelf life. The objective of this study was to examine the effect of temperature and sperm concentration on the in vitro sperm quality during liquid storage for 48 h, including sperm motility and kinetics, response to oxidation, mitochondrial membrane potential (MMP) and DNA fragmentation in goats. Three experiments were performed. In the first, the effects of liquid preservation of semen at different temperatures (5 &deg, C or 17 &deg, C), durations (0, 24 and 48 h) and sperm concentrations (250 &times, 106 sperm/mL (1:2 dilution rate), 166.7 &times, 106 sperm/mL (1:3 dilution rate) or 50 &times, 106 sperm/mL (1:10 dilution rate)) on sperm motility and kinetics were studied. In the second experiment, the effect of temperature, sperm washing and concentration on sperm motility and DNA fragmentation was studied. Finally, the effect of sperm concentration and duration of storage at 5 &deg, C on sperm motility, response to oxidative stress and MMP was examined. We found that refrigerated liquid storage of goat sperm impaired sperm quality, such as motility, MMP and response to oxidation, as storage time increased, however, sperm DNA fragmentation index was not significantly affected. Liquid storage at 5 &deg, C preserved higher total motility than at 17 &deg, C. Moreover, we observed that the reduction of sperm concentration below 500 &times, 106 sperm/mL did not seem to improve the quality of spermatozoa conserved in milk-based extender in the conditions tested.

The role of heat shock protein in reproduction is widely known as a molecular chaperone in aiding and repairing protein formation when stress occurred. The present objectives were to evaluate the effect of different thawing temperature and time on the expression of HSP70 gene expression and the capacitation status in cryopreserved bovine spermatozoa. Briefly, fresh ejaculates were obtained from three different adult bulls. The semen then underwent a sperm washing technique known as Magnetic Activated Cell Sorting System (MACS) and later on, cryopreserved. The sperm- containing straws were then thawed at five different thawing temperatures and time post-cryostorage; 20°C for 13 s, 37°C for 30 s, 40°C for 7 s, 60°C for 6 s and 80°C for 5 s. The RNA was extracted from each of the sperm’s pellets and converted to cDNA prior to the qPCR process. Capacitation status was then determined by means of CTC assay. The results showed that after the process of amplification, there is a significant different of HSP70 gene expression in MACS process samples when the thawing process was performed at 37°C for 30 s, with p

The aim of this study was to test the effects of five different concentrations (0, 10−3, 10−4, 10−5, and 10−6 M) of resveratrol (Res) supplementation in bull sperm washing and fertilisation medium on levels of reactive oxygen species (ROS), phosphatidylserine (PS) externalisation, mitochondrial membrane potential (Δψm), ATP and malondialdehyde (MDA), acrosomal integrity, blastocyst rate, and blastocyst quality after in vitro fertilisation (IVF). The results for sex-sorted sperm from three bulls showed: (1) ROS and MDA levels in 10−3 M and 10−4 M Res groups were significantly lower than those of controls (P −3 M and 10−4 M Res groups were significantly higher than those of controls (P −4 M Res group were significantly higher than those of controls (P −4 M Res supplementation in washing and fertilisation medium of sex-sorted bull sperm significantly decreased ROS, PS externalisation, and MDA, and protected mitochondrial function and acrosomal integrity, thereby increasing blastocyst percentage and quality following IVF.