Ejaculated Spermatozoa Research Articles

The role of testicular sperm extraction in repeated intracytoplasmic sperm injection failures using ejaculate sperm

The role of testicular sperm extraction in repeated intracytoplasmic sperm injection failures using ejaculate sperm

GENETIC POLYMORPHISMS OF CRISP2 GENE IN ASSOCIATION WITH INFERTILITY IN IRAQI PATIENTS

The objective of this study was to evaluate the association and effect of CRISP2 variants on the risk of asthenozoospermia, a male infertility condition marked by absent or diminished sperm motility. There are numerous reasons why individuals develop asthenozoospermia. Therefore, it is crucial to understand the molecular mechanisms underlying this condition of infertility. Furthermore, seminal plasma, a rich source of sperm quality-related biomarkers, transports the many spermatozoa pools that make up human ejaculate down the epididymis. These spermatozoa vary in size, shape, and motility. The morphology and mobility of male ejaculated spermatozoa are affected by a number of genes, including CRISP2. In seminal samples from 120 Iraqi infertile male patients and 40 healthy males who were matched for age, gender, and ethnicity as a control group, the connection of the CRISP2 gene single nucleotide polymorphisms L56V, M176I, and C196R with infertility was investigated. According to statistical analysis of the genotype distribution of these three nsSNPs of the CRISP2 gene in patients with the asthenozoospermia subgroup and the control group, there weren't detectable differences in genotype distribution between AS, OAS, OTA, and fertile men in the Iraqi research sample. Based on allele frequencies, C, T, and G were determined to be protective alleles, with OR values of 0.74, 0.64, and 2.0, respectively.

P-143 Will Use of Testicular sperm in ICSI cycles increase Embryo Aneuploidy?

Abstract Study question Will use of surgically retrieved Testicular Sperms (TESA) in Intra-Cytoplasmic Sperm Injection (ICSI) cycles increase embryo aneuploidy? Summary answer Use of TESA sperms for ICSI seems safe and doesn’t increase embryo aneuploidy. TESA performed for various indications also showed no variation with embryo aneuploidy. What is known already Invention of ICSI and improved surgical sperm retrieval techniques have helped men with severe male factor infertility to father their own genetic child. Sperms retrieved from seminiferous tubules of testes are considered immature as they haven’t completed the process of spermiogensis. It is hypothesized that use of TESA sperm for ICSI can increase embryo aneuploidy, which is still uncertain. Study design, size, duration This retrospective study conducted at a private fertility clinic from 2014–2022 (n = 265 patients; total of 860 embryos evaluated). Study populationTESA sperm used and PGT-A done (n = 66). TESA group was further sub-divided into 2 groups based on the indication for TESA,Non-Obstructive Azoospermia(n = 26) and Raised SDF(n = 40). Control Population Ejaculated sperm used and PGT-A done (n = 199) Only younger women(<37yrs) with self-gametes and RIFconsidered in this study. RIF was defined as women with 2 failed IVFcycles in thepast with at least 4 blastocystsbeing transferred. Participants/materials, setting, methods All fertilized oocytes inseminated by ICSI and subjected to extended blastocyst culture. TESA done as per our clinic’s SOP and for control group sperm preparation done by density gradient method. Trophectoderm biopsy was performed and sent for PGT-A to the genetic lab. Next-Generation sequencing (NGS) was the Comprehensive Chromosomal technique (CCS) used for assessing embryo ploidy status. Aneuploidy across all groups compared. Main results and the role of chance Following were the Aneuploidy rates of TESA sperm, Ejaculate sperm group and TESA done for NOA and SDF respectively. Ejaculate Sperm – Aneuploidy%- 42% TESA Sperm (irrespective of the indication)- Aneuploidy%- 44% (p = 0.6260) TESA Sperm (done for NOA)- Aneuploidy%- 44% (p = 0.7504) TESA Sperm (done for raised SDF) – Aneuploidy%- 32% (p = 0.05) Embryo aneuploidy was comparable between Ejaculate and TESA sperm group. Though TESA sperm is considered immature, it didn’t seem to increase embryo aneuploidy. Embryo Aneuploidy from TESA sperm ICSI cycles done for NOA or raised SDF also showed no altering trend. Data from this study is suggestive that TESA sperm doesn’t alter embryo ploidy status. Limitations, reasons for caution Data is retrospective with small sample size and unequal distribution. A well designed Randomized Trial might help test the inference of this study further. Wider implications of the findings Incidence of male infertility and use of ICSI is on a rise globally. Interventions to optimize embryo implantation and reproductive outcomes will help with better success rates and shorten duration to conception. Further research is warranted with severe male factor infertility to improve sperm selection techniques and reproductive outcomes. Trial registration number not applicable

P-711 Are there any differences among blastocyst euploid and mosaicism rate (high or low) using fresh or frozen ejaculate sperm in donated oocytes?

Abstract Study question To compare euploid and mosaicism rate (high or low) among blastocysts derived from fresh and frozen ejaculate sperm in donated oocytes. Summary answer No statistically significant differences were found on blastocyst euploid and mosaicism rate between blastocysts derived from fresh versus frozen. What is known already Even though 5%-10% autosomal aneuploidies and 5%-100% sex chromosomes aneuploidies are paternally derived, the clinical relevance of paternal contribution to chromosome aneuploidy is frequently overlooked. Besides, male infertility could contribute to the generation of chromosomal abnormalities in the resulting embryos. Sperm cryopreservation is a widely used technique in assisted reproduction for different reasons. However, little is known regarding the potential effect of sperm freezing on embryo aneuploidies. Study design, size, duration Retrospective, observational and multicenter study of patients undergoing Preimplantation Genetic Testing for Aneuploidies (PGT-A) from January 2017 to December 2022 in IVIRMA Madrid, Valencia and Barcelona. To elucidate the effect of advance maternal age, 1,780 biopsied blastocysts coming from the egg donation program were analyzed. Autologous sperm samples with less than 2 million spermatozoa per milliliter and male partners with altered karyotype, abnormal spermatozoa fish or any genetic disease were excluded from the study. Participants/materials, setting, methods Embryos were cultured in regular incubators, with an atmospheric concentration of 5% O2, 6.5% CO2 and a temperature of 37ºC. Blastocyst biopsy was performed either on day 5 or day 6 of embryo development and analyzed through Next Generation Sequencing (NGS). Sperm freezing was done following manufacturer’s instructions (Sperm freeze, Origio). Continuous variables were compared with a student´s t-test and categorical variables with a Chi-square using SPSS statistical program. Main results and the role of chance Mean paternal age (44.3±5.20 years old vs 43.2±5.20 years old), sperm concentration (46.67±33.05 mill/ml vs 35.1±32.92 mill/ml) were not significantly different among fresh versus frozen sperm group (p > 0.05). A total 1780 blastocysts tested for PGT-A were included: 1267 blastocysts coming from fresh ejaculate sperm (n = 194 patients) and 513 blastocysts from frozen sperm (n = 81 patients). No statistically significant differences were found in blastocyst euploid rate between fresh versus frozen ejaculate sperm (64.5% versus 67.2%, p = 0.278 ;) respectively. Blastocyst mosaicism rate: High mosaicism (>50% of aneuploidy trophectoderm cells) and low mosaicism (30-50% of aneuploidy trophectoderm cells) was comparable between fresh versus frozen sperm (2.7% versus 4.5%; p = 0.90) and (3.9% versus 3.7%), respectively. Limitations, reasons for caution This study is based on a retrospective data. Wider implications of the findings According to our findings, there are no statistically significant differences in the blastocyst euploid and mosaicism rate between fresh and frozen ejaculate sperm. Trial registration number Not Applicable

Double semen collection at a 1-h interval in dogs decreases the bacterial contamination of canine ejaculates

Semen extenders usually contain antibiotics with the aim to minimize bacterial growth, but the indiscriminate use of antibiotics increases the emergence of multidrug-resistant bacteria. A limiting factor of semen processing in dogs is the low total sperm count that limits the number of insemination doses that can be obtained from one ejaculate. Therefore, two ejaculates collected at a short interval can be combined to increase the number of AI doses. In this study, semen was collected from dogs either once or the same dogs (n = 28) were submitted to dual semen collection 1 h apart. All ejaculates were submitted to bacteriological analysis. We hypothesized that bacterial contamination of semen is low but that a dual semen collection might increase contamination. A sample for bacteriological examination was taken from raw semen immediately after semen collection. Bacteria including mycoplasmas were isolated using conventional cultivation procedures and isolates were identified to the species level by matrix-assisted laser desorption ionization – time of flight (MALDI-ToF) mass spectrometry. In total, 22 bacterial species were identified in the 84 ejaculates with Mycoplasma cynos, Streptococcus canis and Canicola haemoglobinophilus being most frequent. Bacterial growth was sporadic in 16 and absent in 10 ejaculates. The overall bacterial growth was lower in the second than in the first ejaculate of dual semen collections (p < 0.05). The percentage of motile and membrane-intact spermatozoa in frozen-thawed ejaculates was not associated with the degree of bacterial contamination of raw semen. In conclusion, there was only limited microbial contamination in dog semen and the microorganisms isolated are considered part of the normal genital bacterial flora. Repeated semen collection reduced bacterial contamination in the second in comparison to the first ejaculate. The use of antibiotics in canine semen should be questioned.

Effect of testicular degeneration on expression of sperm protein at 22kDa in stallions.

Sperm protein at 22kDa has been associated with fertility. The objectives of this study were to determine (1) the localization pattern of SP22 on ejaculated and caudal epididymal equine spermatozoa and in epididymal fluid, and to (2) characterize SP22 protein and mRNA expression in testicular and epididymal tissues in response to heat-induced testicular degeneration. Semen was collected before and after hemi-castration, as well as prior to and following insulation of the remaining testes, and tissue specimens were collected for analysis. Histopathology confirmed degeneration in insulated testes. Ejaculated and epididymal spermatozoa from samples collected prior to insulation of the testicles had a predominant staining pattern of SP22 over the equatorial region. However, the equatorial pattern in the pre-insulation epididymal semen samples was significantly lower than in the pre-insulation ejaculated semen samples (68±3, 81±2.6, respectively). Ejaculated and epididymal samples collected after insulation of the testicles showed a complete loss of staining as the predominant pattern. Western blot analysis verified the presence of SP22 on fresh ejaculated spermatozoa prior to and following heat-induced degeneration, on epididymal spermatozoa after testicular insulation, and in testicular and epididymal tissues. Heat insulation significantly reduced messenger RNA expression in the head of the epididymis and testicular tissues. Immunohistochemistry of the testicular and epididymal tissues pre-heating showed considerably weaker staining than the same tissues post-heating. It was concluded that heat-induced testicular damage causes both loss and relocation of SP22 on the sperm membrane. Future studies are warranted to determine the diagnostic value of these findings.

Substitution of soyabean meal by faba bean improves body weight, sperm production and testosterone concentration in mature Queue Fine de l'Ouest rams.

The current study aimed at investigating the effects of partial or total substitution of soybean meal with faba bean on reproductive parameters of Queue Fine de l'Ouest rams. Eighteen adult rams (49.8 ± 3.7 kg and 2.4 ± 1.5 years old) were divided into three homogenous groups. Rams received oat hay ad libitum and 3 types of concentrate (33 g/BW0.75) where soybean meal was the main protein source (SBM diet, n = 6), partially (50%, SBMFB diet, n = 6) or totally (100% FB diet, n = 6) substituted in nitrogen basis by local faba bean. The volume of ejaculate, sperm concentration and sperm mortality rate were measured weekly by collecting semen using an artificial vagina. Serial blood samples were collected 30 days and 120 days after the beginning of the experiment in order to assess plasma testosterone concentrations. The results showed that hay intake was affected (P < 0.05) by the nature of the nitrogen source incorporated (1032.3 ± 12.2; 1026.8 ± 56.6 and 972.8 ± 39.05 g DM/d for SBM, FB and SBMFB respectively. The average live weight of the rams increased from 49.8 ± 0.4 (week 1) to 57.3 ± 0.9 kg (week 17) without effect of the diet. A positive effect of faba bean incorporation in the concentrate was observed on ejaculate volume, concentration and spermatozoa production. All these parameters were significantly higher in SBMFB and FB groups than in SBM group (P < 0.05). The protein source did not affect the percentage of dead spermatozoa and the total abnormalities which were similar for the three diets (38.7; 35.8 and 38.1% for SBM; SBMFB and FB, respectively). The mean testosterone concentration was higher (P < 0.05) in rams fed with faba bean (1.7 ± 0.7 to 1.9 ± 0.7 ng/ml for the SBMFB and the FB groups respectively) than in rams fed SBM diet (1.06 ± 0.5 ng/ml). It was concluded that the substitution of soybean meal by faba bean improved the reproductive performance and did not affect sperm quality in Queue Fine de l'Ouest rams.

(256) Sexual Function and Andrology Parameters in Men after Treatment of Anterior Urethral Strictures with the Optilume Drug Coated Balloon

Abstract Introduction The Optilume Urethral Drug Coated Balloon (DCB) is a novel endoscopic treatment for recurrent male anterior urethral strictures. An important topic of discussion with patients during counseling on treatment options includes the potential affect of treatment on sexual function and fertility. The ROBUST III study was a prospective, randomized study comparing standard of care endoscopic therapy against treatment with the Optilume DCB. Assessments included evaluation of sexual function and andrology parameters. Objective The objective was to study the potential affect of treatment with the Optilume DCB on sexual function and, for younger patients, fertility. Methods A total of 79 men were randomized to receive treatment with the Optilume DCB. Sexual function was assessed via the 15 item International Index of Erectile Function (IIEF) questionnaire. Erectile function, orgasmic function, and overall satisfaction sub-domains were assessed at 3, 6, and 12 months after treatment in patients that were sexually active at baseline. Fifteen additional subjects were enrolled in a non-randomized pharmacokinetic sub-study that included assessments of paclitaxel concentration in semen and sperm quality parameters through 6 months post-treatment. Results Average erectile function and overall satisfaction scores improved from baseline to 12 months post-treatment, while orgasmic function was not changed (Figure). The improvement in satisfaction was statistically significant at 12 months (p=0.003). No adverse events of de novo erectile or ejaculatory dysfunction were reported. Average ejaculate sperm concentration did not change from baseline to 6 months (49 million/mL vs 46.6 million/mL, p=0.88), nor did total spermatozoa per ejaculate (107.2 million vs 104.0 million, p=0.94). Paclitaxel concentration in semen after treatment was maximal at 30 days (2.99 ng/mL) and was reduced near the limit of quantitation of the analytical method by 6 months (0.12 ng/mL). Conclusions Treatment of male anterior urethral strictures with the Optilume DCB did not appear to have an impact on sexual function as measured by the IIEF and adverse event assessments. Average sperm quality parameters did not differ from baseline to 6 months post-treatment. Low levels of paclitaxel were detected in semen after treatment but were nearly undetectable by 6 months. Disclosure Yes, this is sponsored by industry/sponsor: Urotronic Inc. Clarification Industry funding only - investigator initiated and executed study. Any of the authors act as a consultant, employee or shareholder of an industry for: Boston Sci, Coloplast, Medtronic USA Inc., PROCEPT BioRobotic Corporation, NeoTract Inc., Janssen Biotech Inc., Endo Pharmaceuticals Inc., Tolmar Pharmaceuticals, Inc., 180 Medical Inc., AbbVie Inc., Pfizer Inc., Bluewind Medical, Astellas Pharma US Inc., Antares Pharma Inc., Axonic Modulation Technologies Inc., Allergan Inc., Terumo Medical Corporation, Dendreon Pharmaceuticals llc, Blue Earth Diagnostics Limited, Ferring Pharmaceuticals Inc., Olympus America Inc., Karl Storz Endoscopy-America, Clovis Oncology Inc., Dornier MedTech America Inc., Travere Therapeutics Inc., Valeant Pharmaceuticals North America LLC

154 Growth, Libido, Semen Quality, and Reproductive Organ Weights of Hair Sheep Rams Rendered Short-Scrotum at Birth or Weaning

Abstract Rendering male lambs short-scrotum interferes with testicular development and thermoregulation, and has been proposed as a management tool to control male reproduction. This study evaluated at the effect of timing of the procedure (birth or weaning) on growth rate, testis size, libido, ejaculate characteristics and carcass traits in male landrace hair sheep lambs. April- (n = 31) and August-born (n = 23) Barbados Blackbelly (BB) and St. Croix (SC) male hair sheep lambs, balanced by breed and birth type, were rendered short-scrotum (SS) using elastrator bands at birth (B) or weaning (W), or left intact (I). Lambs were born on pasture, weaned at 9 weeks, transitioned in pens for two months post weaning, and then rotationally grazed as a single group to a target weight of 40 kg. At 6-mo of age (peri-pubertal), and again at target weight (post-pubertal), mating behavior was recorded (day) and ejaculate samples collected (day 2). Mating behavior was recorded in a0-min test with males individually exposed to estrus-induced ewes pen. An initial attempt was made to collect semen using an estrus-induced teaser ewe, and if not successful, the semen sample was collected by electro-ejaculation. At target weight lambs were harvested, scrotal content collected, and carcass characteristics (April cohort) determined. Data were analyzed in a model with sex class, breed and cohort as main effects. Pen and pasture ADG (142 and 129 g/d, respectively) and age at target weight (287 d) was not affected (P &amp;gt; 0.1) by SS treatment. However, April-born lambs grew 19 g/d less (P &amp;lt; 0.01) and reached target weight 40 d later (P &amp;lt; 0.001) than August-born males. As for breed, STX had greater pasture ADG (139 vs 124 g/d; P &amp;lt; 0.05) and reached target age 19 days earlier (P&amp;lt; 0.05) than BB males. Libido (number of mounts and services) was not affected by SS treatment, breed or cohort. The frequency of ejaculates containing sperm was less (P &amp;lt; 0.001) in SS (26 to 33%) than intact males (97%) and increased with age in SS but not I males. Sperm concentration and motile sperm in ejaculates were greater (P&amp;lt; 0.05) in I than SS males. Testis and epididymis were larger (P &amp;lt; 0.001) in I (300 and 44.3 g) than in B (134 and 20.0 g) and W (154 and 23.8 g), which were not different. Cohort and breed had no effect (P &amp;gt; 0.1) on mating behavior, ejaculate characteristics, and organ weights. Results suggest that SS and its timing had no effect on growth rate or on mating behavior. Though the procedure depressed the presence of sperm in the ejaculate and decreased testis weight, the suppression was not complete, and caution is warranted in utilizing the technique to facilitate co-grazing ewe lambs.

An Algorithm to Predict the Lack of Pregnancy after Intrauterine Insemination in Infertile Patients.

Increasing intrauterine insemination (IUI) success rates is essential to improve the quality of care for infertile couples. Additionally, straight referral of couples with less probability of achieving a pregnancy through IUI to more complex methods such as in vitro fertilization is important to reduce costs and the time to pregnancy. The aim of the present study is to prospectively evaluate the threshold values for different parameters related to success in intrauterine insemination in order to provide better reproductive counseling to infertile couples, moreover, to generate an algorithm based on male and female parameters to predict whether the couple is suitable for achieving pregnancy using IUI. For that, one hundred ninety-seven infertile couples undergoing 409 consecutive cycles of intrauterine insemination during a two-year period were included. The first year served as a definition of the parameters and thresholds related to pregnancy achievement, while the second year was used to validate the consistency of these parameters. Subsequently, those parameters that remained consistent throughout two years were included in a generalized estimating equation model (GEE) to determine their significance in predicting pregnancy achievement. Parameters significantly associated with the lack of pregnancy through IUI and included in the GEE were (p < 0.05): (i) male age > 41 years; (ii) ejaculate sperm count < 51.79 x 106 sperm; (iii) swim-up alkaline Comet > 59%; (iv) female body mass index > 45 kg/m2; (v) duration of infertility (>84 months), and (vi) basal LH levels > 27.28 mUI/mL. The application of these limits could provide a pregnancy prognosis to couples before undergoing intrauterine insemination, therefore avoiding it in couples with low chances of success. The retrospective application of these parameters to the same cohort of patients would have increased the pregnancy rate by up to 30%.

Unilateral Versus Bilateral Vasoepididymal Anastomosis for Idiopathic Obstructive Azoospermia: A Randomised Controlled Trial

BackgroundMen with idiopathic obstructive azoospermia (OA) are candidates for surgical reconstruction with a vasoepididymal anastomosis (VEA) performed on one or both testis. There are no randomised trials comparing the success of unilateral versus bilateral VEA. ObjectiveWe conducted a randomised trial to compare the two surgical options. Design, setting, and participantsBetween April 2017 and March 2022, men with infertility due to idiopathic OA were randomised to a unilateral (group 1) or bilateral (group 2) VEA in an ethics committee–approved clinical trial, registered with the Clinical Trials Registry. Outcome measurements and statistical analysisThe primary outcome was successful surgery, defined as appearance of sperm in the ejaculate, evaluated at 3 mo intervals after surgery. Additional outcomes were pregnancy rates and complications between the two groups. Men with successful surgery were compared with those without patency to identify the predictors of success. Results and limitationsFifty-four men fulfilled the criteria and 52 who completed follow-up were included in the analysis. The overall patency rate was 36.5% (19/52 individuals). This was higher in men with bilateral surgery (12/26 patients, 46%) than in those with unilateral surgery (7/26 patients, 27%) but was not statistically significant (p = 0.1). The overall pregnancy rate with ejaculated sperm was significantly higher in the bilateral surgery group (4 vs 0, p = 0.037), while the spontaneous conception rate was higher but not statistically significant (3 vs 0, p = 0.074). The complication rates in the two groups were similar (p = 0.7), and all complications were Clavien-Dindo grade 1. Although bilateral surgery and presence of sperm in epididymal fluid were higher in men with patency, these were not statistically significant. ConclusionsA bilateral VEA was associated with higher patency and spontaneous pregnancy rates than unilateral surgery, but the results were not statistically significant. However, the overall pregnancy rate with ejaculated sperm, spontaneous and assisted, was significantly higher in the bilateral surgery group. Patient summaryIn this study, we compared between unilateral and bilateral reconstructive surgery in azoospermic men and found better overall success with bilateral surgery. However, these results were not statistically significant.

DIETARY LEVELS OF PINEAPPLE PEELS ON GROWTH PERFORMANCE, REPRODUCTIVE PERFORMANCE, NUTRIENT DIGESTIBILITY AND CARCASS CHARACTERISTICS OF WEANER RABBITS

A feeding trial was conducted on weaner rabbit (Oryctolagus cunniculus) to assess the growth, reproductive performance, nutrient digestibility and carcass characteristics rabbits fed graded levels of inclusion of dried pineapple peal meal. Thirty-six (36) composite weaned rabbits of mixed sexed and aged between 6 and 8 weeks were randomly allotted to four dietary treatments group with nine rabbits per treatments. Each treatment had three replicates with 3 rabbits in the ratio of 2 females and 1 male per replicate. The pineapple peal was included in the diet at varying inclusion levels of 0,5,10 and 15 % (forming diets T1, T2, T3 and T4, respectively) T1 was the control diet. The experiment lasted for fourteen (14) weeks. Data were collected on growth parameter (body weight feed intake and conversion ratio.), at the end of the 7 weeks digestibility trial using a total collection method. Reproductive performance trial was also determined and carcass characteristics 8 weeks, 2 rabbits per replicate were slaughtered eviscerated and dressed to obtain carcass information on the carcass, weighted and expressed as percentage live weight. Result revealed non- significant (P&lt;0.05) differences in the values obtained for final weight gain, feed conversion ratio and mortality. Result of nutrient digestibility revealed a significant different (P&lt;0.05) most of the parameters (CP. CF and Ash). Significant (P&lt;0.05) different were also recorded in the semen characteristics (sperm per ejaculate and % live sperm. Also, carcass characteristics were observed to be significantly (P&lt;0.05) different on head, fore limb and hind limb). Result obtained for the experiments revealed significant differences in most of growth parameters, higher values of total weight gains and feed intake were observed in T2(5%) 508.33level of inclusion level with T4(15 %) 485.33 lower values. Results of carcass characteristics revealed significant difference (P&lt;0.05) in back, hind limbs and fore limbs, respectively, with T2 having higher values of 181.60, 23.76, for organ weight of weaner rabbits the kidney revealed significant effects (P&lt;0.05) with T2 (5%) having a higher value 6.00 and lower value of 5.20 at T4(15%) inclusion level. Also, the result of semen characteristics revealed significant difference (P&lt;0.05) on sperm ejaculate and live sperm % with T3(10%) having the highest value 355.00 and lowest value of 0.275.00 at T1(5%). It can therefore be recommended that 5% (T2) of dry peal pineapple meal can be included in the diet of weaner rabbit for optimum growth performance, nutrient rabbit digestibility and carcass characteristics while T4(15%) inclusion level for reproductive

MP30-19 HUMAN SPERM IGF2 mRNA LEVELS INVERSELY CORRELATE WITH EMBRYO GROWTH VELOCITY DURING EARLY STAGE OF EMBRYO DEVELOPMENT

You have accessJournal of UrologyCME1 Apr 2023MP30-19 HUMAN SPERM IGF2 mRNA LEVELS INVERSELY CORRELATE WITH EMBRYO GROWTH VELOCITY DURING EARLY STAGE OF EMBRYO DEVELOPMENT Rossella Cannarella, Scott Lundy, Nina Desai, Sarah Vij, and Aldo E. Calogero Rossella CannarellaRossella Cannarella More articles by this author , Scott LundyScott Lundy More articles by this author , Nina DesaiNina Desai More articles by this author , Sarah VijSarah Vij More articles by this author , and Aldo E. CalogeroAldo E. Calogero More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003258.19AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: IGF2 is a potent growth factor capable of influencing cell division and modulating numerous cellular functions. IGF2 mRNA has been identified in human spermatozoa, but little is known about its function in in early embryo development. The aim of this study is to evaluate whether IGF2 levels in spermatozoa is associated with early growth kinetics of human embryos in couples undergoing ICSI. METHODS: This study was conducted on patients undergoing ICSI treatment with fresh oocytes and fresh ejaculated spermatozoa. Couples with infertility due to suspected female factor (e.g., impaired oocyte quality as in patients with severe endometriosis and premature ovarian failure) were excluded from the study. Following clinical use, a surplus aliquot of the semen sample used for ICSI was used to analyze the levels of IGF2 mRNA. The kinetic parameters of each embryo were correlated with the amount of IGF2 mRNA measured in the same swim-up sperm sample used for ICSI. RESULTS: 16 patients were enrolled and a total of 131 embryos were collected. Patients were divided into two groups based on the βhCG test results. We found no statistically significant difference in IGF2 mRNA levels and embryo morphokinetic parameters between the two groups (Table 1). At the correlation analysis, we found a significant negative correlation between sperm IGF2 mRNA levels and the time of pronuclei (tPN) fading (r= -0.2572, p= 0.0078, n=106 embryos), and between sperm IGF2 mRNA and t2 (r= -0.2368, p= 0.0145, n=106 embryos). In contrast, sperm IGF2 mRNA levels were not significantly correlated with tPN, t3, t4, t5, t6, t7, t8, t9, t morula (tM), t blastocyst (tB), and time of expanded blastocyst (tEB) (Table 1). CONCLUSIONS: These results suggest a role for sperm IGF2 mRNA in the very early embryo development (tPN fading -t2). This evidence may help in identify the etiology of apparently idiopathic couple infertility. Indeed, since prolonged t2 has been associated with a poor chance of embryo development into a viable blastocyst, a low level of sperm IGF2 mRNA level can delay t2 and lead to no blastocyst formation. Source of Funding: None © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e398 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Rossella Cannarella More articles by this author Scott Lundy More articles by this author Nina Desai More articles by this author Sarah Vij More articles by this author Aldo E. Calogero More articles by this author Expand All Advertisement PDF downloadLoading ...

Surgically retrieved spermatozoa for ICSI cycles in non-azoospermic males with high sperm DNA fragmentation in semen.

Intracytoplasmic sperm injection (ICSI)using surgically retrieved spermatozoa outside the classic context of azoospermia has been increasingly used to overcome infertility. The primary indications include high levelsofsperm DNA damage in ejaculated spermatozoa and severe oligozoospermia or cryptozoospermia, particularly in couples with ICSI failurefor no apparent reason. Current evidence suggests that surgically retrieved spermatozoa for ICSI in the above context improves outcomes, mainly concerning pregnancy and miscarriage rates. The reasons are not fully understood but may be related to the lower levels of DNA damage in spermatozoa retrieved from the testis compared with ejaculated counterparts. These findings are consistent with the notion that excessive sperm DNA damage can be a limiting factor responsible for the failure to conceive. Using testicular in preference of low-quality ejaculated spermatozoa bypasses post-testicular sperm DNA damage caused primarily by oxidative stress, thusincreasing the likelihood of oocyte fertilization by genomically intact spermatozoa. Despite the overall favorable results, data remain limited, andmainly concern males with confirmed sperm DNA damage in the ejaculate. Additionally, information regarding the health of ICSI offspring resulting from theuseofsurgicallyretrievedspermatoaofnon-azoospermicmales is still lacking. Efforts should be made to improve the male partner's reproductive health for safer ICSI utilization. A comprehensive andrological evaluation aiming to identify and treat the underlying male infertility factor contributing to sperm DNA damage is essential for achieving this goal.

Does SARS-CoV-2 Affect Human Semen? A Systematic Review and Meta-Analysis

Contradictory results have been reported regarding effects of the SARS-CoV-2 upon human semen. A timely and up-to-date systematic review with meta-analysis appears necessary. This study aimed to deliver pooled prevalence (PP) of SARS-CoV-2 in semen and pooled semen parameters as compared with the uninfected. The relevant databases were scanned by two authors for observational studies reporting analysis of semen in COVID-19 patients. The SARS-CoV-2-infected were assigned to group A (exposed arm), whereas the uninfected to group B (unexposed arm). Newcastle–Ottawa Scale was used to address the risk of bias. PRISMA guidelines were adopted. In case of homogenous studies, fixed-effects model was followed, whereas for heterogenous studies random-effects model was used. Of 990 studies, 24 were eligible involving 1589 subjects (947 in group A and 642 in group B). The “comparability” domain was biased the most. SARS-CoV-2 RNA was detected in three studies among 8 individuals producing the PP of 1.76% (95% CI 0.72–3.21). Sperm concentration was reduced significantly (WMD = –16.23 [95% CI –25.56 to –6.89], as well as total sperm in ejaculate (WMD = –34.84 [95% CI − 43.51 to –26.17]) and sperm volume (WMD = − 0.48 [95% CI − 0.59 to − 0.36] in group A as compared with controls. There was a non-significant effect upon progressive motility and leukocyte presence in semen. SARS-CoV-2 RNA in semen among the infected individuals is detected infrequently. By this token, sexual transmission through semen is of low probability and little concern for public health. However, significant decrease in sperm volume, sperm concentration, and total sperm in ejaculate has been noted. The current data, though, are limited, and more studies with longer follow-up are needed to evaluate the further impact.

Genetic background of semen parameters in Italian Simmental bulls

The aim of this study was to estimate genetic parameters and investigate the genomic background of scrotal circumference and semen parameters in the Italian Simmental bulls. Scrotal circumference, number of normal spermatozoa, ejaculate volume, spermatozoa motility, and total spermatozoa were measured on 622 bulls, of which 603 had genotypes for 42,141 SNP. Variance components of scrotal circumference were estimated with an animal model that included the fixed effects of birth year, animal age, and measurement method, and the random effects of day of measurement and animal. In the model for the other traits, the scrotal circumference was added as a covariate to account for its influence on the semen parameters. A genome-wide association study was carried out using the ssGBLUP-approach. Heritabilities ranged from 0.07 ± 0.05 (spermatozoa motility) to 0.50 ± 0.14 (scrotal circumference). A total of 13 SNP passed the Bonferroni correction threshold and the number of significantly associated markers ranged from 1 (ejaculate volume and spermatozoa motility) to 5 (total spermatozoa). Genes already associated with reproduction parameters were retrieved close to the significant SNP. Results of the present study gave preliminary insights about the genetic determinism of semen quality in Italian Simmental bulls. Highlights Low to moderate heritabilities were estimated for semen traits. Few markers were associated with the phenotypes, suggesting their polygenic determinism. Candidate genes already associated in literature with sperm traits were found.

Location of aquaporins 3, 7 and 10 in frozen-thawed ejaculated and cauda epididymal spermatozoa from the Iberian ibex, mouflon, and chamois

Spermatozoa collected from the cauda epididymis of wild ruminants are more cryoresistant than ejaculated spermatozoa. Changes in the membrane location of aquaporins (AQPs) follow the osmotic changes that occur during freeze-thawing, and might influence the cryosurvival of spermatozoa depending on their source. This work reports the location of AQP3, AQP7 and AQP10 in the cauda epididymal and post-ejaculation spermatozoa of three wild mountain ungulate species (Iberian ibex, mouflon, and chamois), as determined by Western blotting (WB) and immunocytochemistry (ICC) using commercial rabbit polyclonal primary antibodies. WB confirmed the presence of all three AQPs in the spermatozoa of all the studied species, while ICC showed AQP3 to be mainly located in the sperm acrosome, mid-piece, principal piece, and end piece, both in cauda epididymal and ejaculated cells. The percentage of ejaculated spermatozoa showing AQP3 in the principal piece was higher in the ibex than in the chamois (P < 0.05), and higher in epididymal spermatozoa in the mouflon than in the chamois (P < 0.05). AQP7 was located in the acrosome of both epididymal and ejaculated spermatozoa, as well as in the cytoplasmic droplet of the epididymal spermatozoa of all three species. No differences were seen between the species with respect to the percentage of spermatozoa showing AQP7. AQP10 was located mainly in the mid-piece, principal piece and end piece of the sperm tail in both epididymal and ejaculated spermatozoa. The percentage of mouflon spermatozoa with AQP10 in the end piece was higher in the cauda epididymal than in the ejaculated spermatozoa (P < 0.05). In conclusion, except for AQP10 in the mouflon, the locations of the studied AQPs are similar in epididymal and ejaculated spermatozoa, with inter-species differences seen only for AQP3. Further studies are needed to determine what this might mean with respect to sperm cryopreservation.

THE REPRODUCTIVE ABILITY OF MALE RABBITS DEPENDING ON THE INFLUENCE OF PARATYPICAL AND GENOTYPICAL FACTORS

THE REPRODUCTIVE ABILITY OF MALE RABBITS DEPENDING ON THE INFLUENCE OF PARATYPICAL AND GENOTYPICAL FACTORS

Female factors modulate Sex Peptide’s association with sperm in Drosophila melanogaster

BackgroundMale-derived seminal fluid proteins (SFPs) that enter female fruitflies during mating induce a myriad of physiological and behavioral changes, optimizing fertility of the mating pair. Some post-mating changes in female Drosophila melanogaster persist for ~10–14 days. Their long-term persistence is because the seminal protein that induces these particular changes, the Sex Peptide (SP), is retained long term in females by binding to sperm, with gradual release of its active domain from sperm. Several other “long-term response SFPs” (LTR-SFPs) “prime” the binding of SP to sperm. Whether female factors play a role in this process is unknown, though it is important to study both sexes for a comprehensive physiological understanding of SFP/sperm interactions and for consideration in models of sexual conflict.ResultsWe report here that sperm in male ejaculates bind SP more weakly than sperm that have entered females. Moreover, we show that the amount of SP, and other SFPs, bound to sperm increases with time and transit of individual seminal proteins within the female reproductive tract (FRT). Thus, female contributions are needed for maximal and appropriate binding of SP, and other SFPs, to sperm. Towards understanding the source of female molecular contributions, we ablated spermathecal secretory cells (SSCs) and/or parovaria (female accessory glands), which contribute secretory proteins to the FRT. We found no dramatic change in the initial levels of SP bound to sperm stored in mated females with ablated or defective SSCs and/or parovaria, indicating that female molecules that facilitate the binding of SP to sperm are not uniquely derived from SSCs and parovaria. However, we observed higher levels of SP (and sperm) retention long term in females whose SSCs and parovaria had been ablated, indicating secretions from these female tissues are necessary for the gradual release of Sex Peptide’s active region from stored sperm.ConclusionThis study reveals that the SP-sperm binding pathway is not entirely male-derived and that female contributions are needed to regulate the levels of SP associated with sperm stored in their storage sites.

Appraisal of ATP1B1 and GSTM3 proteins as freezability factors in buffalo ejaculated spermatozoa

Appraisal of ATP1B1 and GSTM3 proteins as freezability factors in buffalo ejaculated spermatozoa