Total Sperm Research Articles

EFFECT OF EXOGENOUS GONADOTROPHIN-RELEASING HORMONE ON SEMEN CHARACTERISTICS AND LIBIDO OF RABBIT BUCKS

Twenty-seven (27) New Zealand White rabbit bucks aged 6 — 8 were used to evaluate the effect of Ovaprim, an exogenous gonadotropin releasing hormone (GnRH), on their seminal characteristics and libido. The rabbits were randomly selected and assigned to three treatment groups designated as Tl, T2 and T3 in a completely randomized design (CRD). The exogenous hormone was administered by intramuscular injection at the rate of 0.00ml, 0.20ml and 0.40ml per kilogram body weight for T1, T2 and T3 respectively, with Tl serving as the control experiment. Semen samples were collected at the beginning of the experiment to determine the baseline and after 48 days of administration of the hormone. Libido was measured 3 hours after administering the GnRH. Results obtained from the baseline study showed no significant difference (P>0.05) in the various treatments for semen volume, mass motility, sperm concentration, live sperm proportion, number of spermatozoa per ejaculate, total viable spermatozoa, normal sperm proportion, semen viscosity, semen colour, libido score and reaction time. At the end of the study, semen volume, sperm concentration, live sperm proportion, number of spermatozoa per ejaculate, total viable sperm, semen viscosity and libido were found to differ significantly (P<0.05) between the treatments. It was concluded from this study that the administration of 0.2ml/kg exogenous GnRH (Ovaprim) used for this experiment led to enhanced reproductive performance with respect to libido and semen quality and had little or no deleterious effect on the rabbit bucks.

SEMEN CHARACTERISTICS AND SCROTAL BIOMETRY OF WEST AFRICAN DWARF RAMS FED AMMONIUM SULPHATE FORTIFIED DIET

Ammonium sulphate (AS) is non-protein nitrogen sources widely used as a feed additive for ruminants to provide sulphur (S) to the diet. This study assessed the semen characteristics and scrotal biometry of West African dwarf rams fed AS fortified diets at 0. 2.5, 5.0 and 7.5g/kg to the concentrate diet as T1, T2, T3 and T4 respectively. Sixteen WAD rams with average body weight of 12.20+0.12 kg were used and fed for 105 days. Each treatment had four replicates while semen was collected once from all replicates in the treatments. There was significant difference in the sperm concentration (x 109/mL) with T4 (160.06) having the highest while T1 (136.00) had the lowest. The total sperm ejaculate followed the same trend as sperm concentration with the lowest value recorded for rams fed T1 (98.24) while those on T4 (150.66) had the highest value. There were significant differences in the values recorded for the scrotal circumference and length. however, scrotal width, testicular weight, scrotal length and scrotal circumference values in T4 was numerically higher than other treatments. It can be concluded that AS fortified diets at 7.5g/kg improved the semen quality positively.

Sperm Quality in 1252 Adolescents and Young Adults (AYAs) Undergoing Fertility Preservation Due to Cancer or Nonmalignant Diseases.

Purpose: To investigate the quality of emergency-collected semen samples aimed at sperm cryopreservation provided by adolescents and young adults (AYAs) presenting with cancer or nonmalignant diseases. Methods: This is a prospective cohort study of postpubertal males referred for sperm cryopreservation who provided at least one semen sample for fertility preservation at the Reproductive Medicine Clinic of Karolinska University Hospital, Stockholm, Sweden, between January 2009 and January 2020. Sperm quality was assessed by total sperm count, concentration, and motility. Sperm quality by disease groups was compared with the reference population data of fertile men defined by the World Health Organization (WHO). Results: Among the 1252 patients who provided samples for cryopreservation, 1063 had cancer and 189 had nonmalignant diseases. The most common malignant indications included testicular cancers (n = 501) and Hodgkin lymphoma (n = 102). Among those with nonmalignant disease, 35% (n = 66) had testicular disease. Sperm quality was significantly lower in all groups of patients with cancer compared with the reference population. In total, azoospermia was found in 8% of the patients with cancer, in 9% of those with nonmalignant testicular disease, and in 3% of the remaining men with nonmalignant disease. Conclusion: Sperm quality in adult patients with cancer was significantly impaired compared with the WHO reference population standards for fertile men. For adolescent patients, standard reference values are lacking. AYAs wishing to preserve fertility should receive individualized counseling regarding sperm quality at the time of cryopreservation, and in selected cases, banking of additional samples should be recommended depending on the sperm quality parameters.

Biogenic and risk elements in wild boar testes and relation to spermatozoa motility

Environmental pollution results in serious health hazards to animals, reflected in biogenic and risk element concentrations in animal tissues. The objective of this study was to examinate concentration of selected elements in testes, and epididymal spermatozoa motility of wild boars. Wild boars were hunted in region Žuhračka - Levice, Slovak Republic. Testes were removed postmortem, spermatozoa were collected from cauda epididymis and assessed by CASA system. Elements concentrations were measured by ICP and by CV-AAS. Spermatozoa motility was 44.29% and progressive motility 18.47%. Concentration of elements in testes was in following order: K > Na > Mg > Ca > Fe > Zn > Al > Cu > Se > Mn > As > Cr > Pb > Mo > Sr > Ni > Ba > Cd > Li > Hg. The most notable correlations indicate association between Se and total spermatozoa motility, as well as with progressive motility, furthermore between As and velocity curved line and beat cross frequency. A high positive significant correlation was found between mercury and beat cross frequency. The data may serve as a fine control indicator to detect potentially toxic elements accumulated from polluted environment that can affect reproduction of wild animals.

Individual and joint associations of air pollutants exposure with semen quality: A retrospective longitudinal study in Wenzhou, China.

The impact of air pollution on semen quality has been confirmed, yet the joint effect remains unclear. We evaluate the individual and joint associations of particulate (PM2.5 and PM10) and gaseous pollutants (NO2, SO2, O3 and CO) with semen quality. We included 5,114 men in this study from 2014 to 2022. The individual and joint associations were measured by multiple linear regression models. Sperm motility and semen volume were inversely associated with pollutant concentrations during every stage of sperm development, especially at lag days 0-9 and 10-14 (all P < 0.05). Stratified analyses showed that the study pollutants (except CO) had a positive effect on semen concentration during the stage of sperm development, especially in spring and autumn, while a decreased total sperm number was associated with CO (all P < 0.05). However, joint associations of particulate and gaseous pollutants with semen quality parameters were not statistically significant (all P > 0.05). During all stages of sperm development, particulate and gaseous pollutants had individual negative impacts on spermmotility andsemen volume, and these impacts were less pronounced in spring and autumn. Our findings highlight the importance and necessity of reducing the exposure to pollutants especially in the critical stage of sperm development to improve semen quality.

Exposome Profiling of Environmental Pollutants in Seminal Plasma and Novel Associations with Semen Parameters.

Indicators of male fertility are in decline globally, but the underlying causes, including the role of environmental exposures, are unclear. This study aimed to examine organic chemical pollutants in seminal plasma, including both known priority environmental chemicals and less studied chemicals, to identify uncharacterized male reproductive environmental toxicants. Semen samples were collected from 100 individuals and assessed for sperm concentration, percent motility, and total motile sperm. Targeted and nontargeted organic pollutant exposures were measured from seminal plasma using gas chromatography, which showed widespread detection of organic pollutants in seminal plasma across all exposure classes. We used principal component pursuit (PCP) on our targeted panel and derived one component (driven by etriadizole) associated with total motile sperm (p < 0.001) and concentration (p = 0.03). This was confirmed by the exposome-wide association models using individual chemicals, where etriadizole was negatively associated with total motile sperm (FDR q = 0.01) and concentration (q = 0.07). Using PCP on 814 nontargeted spectral peaks identified a component that was associated with total motile sperm (p = 0.001). Bayesian kernel machine regression identified one principal driver of this association, which was analytically confirmed to be N-nitrosodiethylamine. These findings are promising and consistent with experimental evidence showing that etridiazole and N-nitrosodiethylamine may be reproductive toxicants.

In Vitro Gene Conservation Status and the Quality of the Genetic Resources of Native Hungarian Sheep Breeds.

Studies revealed a global loss of genetic resources for local sheep breeds. Therefore, the current study aimed to introduce and highlight the progress made on Hungary's existing gene conservation program (small Gene Bank). Furthermore, we evaluated breed (Tsigai, Cikta, and Racka), season, and individual variabilities (n = 24) of the pre-freeze and post-thaw semen stored in the Gene Bank to enhance the gene conservation of the breeds. The samples were cryopreserved manually, and post-thaw spermatozoa were analyzed for motility (CASA), viability, chromatin structure, and morphometry of the sperm nuclei. Ejaculate volume, spermatozoa concentration, subjective motility and standard motility, kinematic parameters, and spermatozoa's head area standard deviation of the post-thaw samples differed significantly among breeds (p < 0.05). Season affected ejaculate volume, total spermatozoa number/ejaculate, STR, BCF, and ALH. We observed a significant (p < 0.001; 0.05) breed and season interaction on concentration, total spermatozoa number/ejaculate, VCL, LIN, WOB, spermatozoa's head average perimeter and nucleus length (Tsigai and Cikta differed but were statistically the same as Racka). Similarly, season significantly (p < 0.05) affected the proportion of ejaculate suitable for freezing. There was a significant (p < 0.05) difference in kinematic parameters and viability among the rams across the breeds. The spermatozoa's head morphometry of the Tsigai and Cikta breeds differed significantly (p < 0.05) among the rams. There were individual and breed differences in many spermatozoa quality parameters. The stored samples are of good quality, with more than 40% having intact membranes and low abnormal chromatin condensation.

Sperm-borne miR-34c-5p and miR-191-3p as markers for sperm motility and embryo developmental competence.

Sperm-borne microRNAs play a pivotal role in influencing essential cellular processes during fertilization, impacting the quality of embryo development. Dysregulated microRNA profiles have been associated with compromised embryonic development and increased incidences of pregnancy loss. This study aimed to investigate the potential associations between the abundance of miR-34c-5p and miR-191-3p in human spermatozoa with sperm quality, as well as with embryo quality and metabolic performance during in vitro development. Thirteen couples who underwent a total of 13 cycles participated in this study. The sperm quality was assessed using conventional methods following World Health Organization guidelines. Quantitative polymerase chain reaction was employed to measure microRNA abundance in spermatozoa. Embryos were categorized as good, lagging, or bad based on morphokinetic evaluation. Evaluation of embryo metabolic performance involved tracking changes in specific metabolites within the cultured media using nuclear magnetic resonance spectroscopy. Statistical analysis was conducted to explore the correlation between microRNA abundance in human spermatozoa and all other collected data. Our findings revealed a negative correlation between the abundance of miR-34c-5p (but not miR-191-3p) and total sperm motility, potentially mediated by the modulation of key signaling pathways. Additionally, higher levels of miR-34c-5p in spermatozoa were strongly associated with the consumption or release of key metabolites by developing embryos, particularly those linked with lipid and glucose metabolism, suggesting enhanced metabolic performance, while miR-191-3p was mostly associated with glucose consumption. Concurrently, only miR-34c-5p content in spermatozoa correlated with higher embryo quality. This study provides evidence suggesting that the abundance of miR-34c-5p in spermatozoa is correlated not only with total sperm motility but also with markers of embryo developmental competence, highlighting the potential significance of this sperm microRNA content as a biomarker in assisted reproduction.

Downward trend in male reproductive health and fertility in Eastern Iran.

This study aimed to assess the ten-year trend in semen quality among couples referred to the Infertility Center in Kerman between 2008 and 2017. The study included 2952 semen samples from men 18 to 60 years old referred to the infertility center as infertile couples living in Kerman province, Iran, whether they had normal or abnormal semen analysis. A total of 2952 sperm samples were included. Statistically significant changes were observed in semen parameters. Particularly, significant changes were observed for volume (-0.08 mL/year), sperm concentration (-2.34 (mio/mL)/year), total sperm count (-13.17 (mio/ejaculate)/year), progressive motility (-2.62%/year), non-progressive motility (-0.59%/year), immotile sperm (2.49%/year), and normal morphology (-0.134%/year). In bivariate analysis, the prevalence of oligozoospermia in this study showed a statistically significant association with age (OR = 1.019; 95% CI = 1.007-1.032; p = 0.003). Likewise, there was a statistically significant association with the year (OR = 1.087; 95% CI = 1.050-1.125; p = 0.000). Semen quality parameters showed a downtrend during the last 10 years in this study, emphasizing the importance of male reproductive health monitoring and warning public health coordinators to pay more attention to this important issue.

Chronic oral administration of L-carnitine induces testicular injury: in vivo evidence.

While L-carnitine is commonly used to treat oligoasthenozoospermia, concerns have been raised regarding its potential harm to spermatogenesis. This study aims to investigate the potential testicular toxicity of long-term oral administration of L-carnitine. In this study, we refer to the clinical adult dosage and mode of L-carnitine administration, and after converting to mouse doses, mice were daily intragastrical administered L-carnitine to investigate whether it was harmful to the testis. The investigation involved assessing its potential testicular toxicity through histopathological staining, sperm motility analysis, and quantitative real-time PCR. Our results showed that L-carnitine increased sperm motility after 14days of continuous administration, but increased luminal exfoliated spermatogenic cells occurred in the testis, and TUNEL results showed increased apoptotic cells. Compared with the control group, the mRNA expression of the spermatogenic cell marker at each stage was decreased in mice treated for 14 consecutive days of L-carnitine. After 50days of continuous administration followed by 14days of drug withdrawal, the total sperm motility of mice was almost 0, and a large number of abnormal eosinophilic spermatogenic cells appeared in the testis. These indicate that oral L-carnitine for more than 14 days impairs spermatogenesis in mice, and sudden discontinuation of administration results in substantial death of established spermatogenic cell populations. Our findings suggest that chronic oral administration of L-carnitine impairs spermatogenic function in the testis. The oral administration of L-carnitine to enhance sperm motility should not exceed the 2/5 point of the spermatogenic cycle.

Pseudoephedrine for ejaculatory dysfunction after retroperitoneal lymph node dissection in testicular cancer.

To assess the impact of ejaculatory dysfunction (EjD; failure of emission or retrograde ejaculation) on health-related quality of life (HRQoL) after retroperitoneal lymph node dissection (RPLND) for testicular cancer and explore the efficacy of pseudoephedrine hydrochloride as treatment. In a single arm, phase II trial, patients at ≥6 months after RPLND were invited to complete patient-reported outcome measures (European Organisation for Research and Treatment of Cancer [EORTC] quality of life questionnaire [QLQ]-30-item core, EORTC QLQ-testicular cancer-26, and Brief Male Sexual Function Inventory) evaluating HRQoL and sexual function in follow-up (ACTRN12622000537752/12622000542796). If EjD was reported, post-ejaculatory urine ± semen analysis was undertaken. In eligible patients, pseudoephedrine hydrochloride 60 mg was administered orally every 6 h for six doses. The primary endpoint was sperm count >39 million sperm/ejaculate (>5th centile) following treatment. The trial was powered to detect a clinically relevant 36% achieving sperm count of >39 million sperm/ejaculate. Secondary endpoints included semen volume >1.5 mL, total motile sperm count, safety, and HRQoL impacts. Of the 58 patients enrolled, the median (interquartile range [IQR]) age was 35 (29-41) years, with a median (IQR) of 37 (18-60) months from RPLND. EjD was reported in 33 (57%), including 27/52 (52%) receiving follow-up at our centre. There were no differences in global HRQoL; however, role functioning (P = 0.045), sexual problems (P < 0.005), and sexual enjoyment (P = 0.005) was poorer if EjD was present. In all, 24/33 (73%) patients with EjD consented to pseudoephedrine treatment. Of 22 evaluable patients, four (18%) achieved a sperm count of >39 million/ejaculate (P = 0.20), and four (18%) had a semen volume of >1.5 mL (P = 0.20). There was a mean increase of 105 million sperm/ejaculate (P = 0.051) and 1.47 mL increase in semen volume (P = 0.01). No safety concerns arose. Ejaculatory dysfunction is common after RPLND but did not impact global HRQoL in our cohort. Pseudoephedrine improved EjD for some; however, its efficacy was lower than expected. Pseudoephedrine may be considered on an individualised basis.

The effect of mixed honeybee drone semen on sperm quality characteristics: A preliminary study

Following copulation, sperm of multiple honeybee (Apis mellifera (A.m.) Linnaeus (Apidae)) drones are mixed inside the spermatheca of a queen, and sperm behaviour in this environment is of importance for sperm survival and successful fertilization. The polyandrous mating system of honeybees further may allow for cryptic female choice and sperm competition to select for specific sperm traits to enable successful reproduction. This preliminary study therefore examined multiple sperm quality parameters in mixed drone semen. Thirty-one drone ejaculates were collected and split to be mixed with one and two other drones’ semen. A computerised sperm analysis system was used for analysing sperm functionality parameters, at baseline and 60 minutes. Morphology measurements included head, tail, and total sperm length. Sperm concentration was significantly higher in three-drone samples compared to other groups (p &lt; 0.001), while comparisons of sperm functional and morphometric parameters did not reveal differences. However, after 60 minutes, mixed semen samples displayed changes in terms of sperm functionality. The motility percentage significantly decreased in two-drone samples (p = 0.004). In contrast, three-drone samples had consistently lower motility and kinematic parameters at baseline, but following an hour it was the only group to show improvement in sperm progressive motility, swimming speed and kinematics. Sperm concentration correlated with the majority of functionality parameters, while, morphological measurements did not. Sperm functionality changes observed in this study, can be attributed to a combination of factors, including, mixing of drone semen, sperm concentration, and time allowing for potential sperm interaction.

Effect of Breed and Season in Buck Semen Cryopreservation: The Portuguese Animal Germplasm Bank.

The aims of this study were to characterize the semen as well as the influence of breed, season, and semen processing on spermatozoa (SPZ) traits of four native Portuguese goat breeds used for the bank of Portuguese animal germplasm (BPAG). A total of 1017 ejaculates from Serrana (n = 30), Bravia (n = 15), Charnequeira (n = 11), and Preta de Montezinho (n = 3) bucks were collected between 2004 and 2020 at (EZN-INIAV; 39° N) during the whole year under natural conditions. All the fresh and cryopreserved (-196 °C) semen was evaluated and stored in the BPAG. Bravia bucks (the smallest breed) produced less (p < 0.05) volume of ejaculate than all the other breeds, which was higher during the full breeding season (September-January; p < 0.05), regarding all the other breeds. Contrarily, in general, SPZ concentration was lower during September-January, but total SPZ per ejaculate remained similar (p > 0.05) during May-August and September-January in Serrana bucks. The SPZ viability and SPZ midpiece defects were slightly influenced by breed and SPZ head defects by season (lowest % in February-April; p < 0.05). On the contrary, the freezing-thawing cycle strongly influenced (p < 0.01) all SPZ traits. The correlation coefficients of these traits between fresh and thawed SPZ were low (up to 0.33; p < 0.01), highlighting the importance of semen processing in semen cryopreservation. We conclude that breed and season had a relevant effect on ejaculate traits, but it was much less evident for the studied SPZ traits. These native goats can serve as semen donors throughout the year, under natural conditions.

Ambient air pollution decreased normal fertilization rate via the mediation of seminal prosaposin

ObjectiveThis study focuses on the association between seminal concentration of prosaposin and ambient air pollutants and whether the association affects the normal fertilization rate in vitro fertilization (IVF) treatment. MethodsThe cohort of 323 couple participants aged 22–46 was recruited from Jan. 2013 to Jun. 2018. At enrollment, resident address information was obtained and semen parameters of male counterparts were evaluated according to WHO criteria. We used inverse distance weighting interpolation to estimate the levels of ambient pollutants (SO2, O3, CO, NO2, PM2.5, and PM10) in the surrounding area. The exposure of each participant was estimated based on the data gathered from air quality monitoring stations and their home address over various periods (0–9, 10–14, and 0–90 days) before semen sampling. The generalized linear regression model (GLM) and the Bayesian kernel machine regression (BKMR) were used to analyze the associations between pollutants, semen parameters, prosaposin, and normal fertilization. Additionally, the mediating effect of prosaposin and semen parameters on the link between pollutants and normal fertilization was investigated. ResultsGLM and BKMR showed exposure to ambient air pollutants was all associated with the concentration of seminal prosaposin, among them, O3 and CO were also associated with normal fertilization (-0.10, 95 %CI: −0.13, −0.06; −26.43, 95 %CI: −33.79, −19.07). Among the semen parameters, only the concentration of prosaposin and total motile sperm count (TMC) was associated with normal fertilization (0.059, 95 %CI: 0.047, 0.071; 0.016, 95 %CI: 0.012, 0.020). Mediation analysis showed that prosaposin played a stronger mediating role than TMC in the relationship between short-term exposure to O3 and fertilization (66.83 %, P<0.001 versus 3.05 %, P>0.05). ConclusionSeminal plasma prosaposin showed a stronger meditating effect reflect the correlation between ambient air pollutants and normal fertilization rate than conventional semen parameters, which may be used as one of the indicators between pollution and fertilization in IVF.

Sperm Size Decreases With Increasing Pheomelanin Pigmentation but Not With the Amount of Glutathione in the Barn Swallow.

Pigment-based coloration is prevalent in animals, but its expression greatly varies across species, populations, and even among individuals in the same populations. Some animals are highly pigmented and thus have conspicuous coloration, whereas others are modestly pigmented and thus have drab coloration. A possible explanation for the variety in pigmentation is a resource-based tradeoff, in which resources invested in pigmentation are unavailable for other functional traits, and thus animals that need to invest in the latter have limited resources to invest in pigmentation. Resource-based tradeoff is plausible in theory, but direct tests are scarce, partially because of many components of pigment-based coloration (i.e., multiple pigments, integument microstructure, and stains) that affect coloration, preventing the use of coloration as an index of pigmentation. Here, using the barn swallow, Hirundo rustica, we examined the relationship between pheomelanin pigmentation in reddish throat patch (a precopulatory sexual trait) and total sperm length (a postcopulatory sexual trait), with particular attention to glutathione as the common resource. We predicted that pheomelanin, which is the predominant pigment in the reddish throat patch, should be negatively related to total sperm length, and that both sexual traits should be further negatively related to the amount of glutathione. As predicted, we found a negative relationship between pheomelanin pigmentation and total sperm length. However, the amount of glutathione in the blood showed no detectable relationship to them. The tradeoff between pheomelanin pigmentation and sperm size, as inferred from the current and previous results, might not be a simple glutathione-based tradeoff.

Comparison of Nanoparticles and Single-Layer Centrifugation for Separation of Dead from Live Stallion Spermatozoa.

The goal of this study was to compare the efficacy of coated iron-core nanoparticles and single-layer centrifugation for separation of dead from live stallion spermatozoa. Our hypothesis was that nanoparticles would bind to dead sperm and allow for separation from live sperm using a magnet, resulting in a population of spermatozoa with a high percentage of total and progressive motility. Treatment Group 1 was an untreated control. Treatment Group 2 (nanoparticles, NP) utilized sperm incubated with nanoparticles followed by application of a magnet to remove dead sperm adhered to the coated nanoparticles. Treatment Group 3 (single-layer centrifugation, SLC) layered sperm above EquiPure™ followed by centrifugation. Semen samples were subsequently evaluated for sperm motility parameters, plasma membrane integrity, acrosome status, and morphology. The SLC technique yielded higher (p < 0.05) progressive motility (76 ± 9.2%) than the NP separation technique (59 ± 12.2%) or the untreated control (47.3 ± 5.1%). However, the total number of sperm recovered was higher (p < 0.05) in the NP technique (526.2 ± 96.6 × 106) than the SLC procedure (211.7 ± 70 × 106), yielding a higher total number of progressively motile sperm (317.6 ± 109 × 106) recovered using the NP technique than the SLC technique (157.8 ± 43.6 × 106). The percentage of live, acrosome intact sperm recovered was higher for SLC than NP. In summary, the SLC technique yielded a higher percentage of sperm motility, intact plasma membranes, and acrosome integrity, but yielded lower total sperm than with the nanoparticle separation technique.

Cr (VI)-induced ribosomal DNA copy number variation is associated with semen quality impairment: Evidence from human to animal study

ObjectivesThis study aimed to analyze the possible role of rDNA copy number variation in the association between hexavalent chromium [Cr (VI)] exposure and semen quality in semen donors and further confirm this association in mice. MethodsIn this cross-sectional study, whole blood and semen samples were collected from 155 semen donors in the Zhejiang Human Sperm Bank from January 1st to April 31st, 2021. Adult C57BL/6 J male mice were treated with different doses of Cr (VI) (0, 10, or 15 mg/kg b.w./day). Semen quality, including semen volume, total spermatozoa count, sperm concentration, progressive motility, and total motility, were analyzed according to the WHO laboratory manual. Cr concentration was detected using inductively coupled plasma mass spectrometry. The rDNA copy number was measured using qPCR. ResultsIn semen donors, whole blood Cr concentration was negatively associated with semen concentration and total sperm counts. Semen 5 S and 45 S rDNA copy numbers were negatively associated with whole blood Cr concentration and whole blood 5.8 S rDNA copy number was negatively associated with semen Cr concentration. In mice, Cr (VI) damaged testicular tissue, decreased semen quality, and caused rDNA copy number variation. Semen quality was related to the rDNA copy number in whole blood, testicular tissue, and semen samples in mice. ConclusionCr (VI) was associated with decreased semen quality in semen donors and mice. Our findings suggest an in-depth analysis of the role of the rDNA copy number variation in the Cr (VI)-induced impairment of semen quality.

O-017 Swim-up versus density gradients for sperm preparation in intrauterine insemination: a multi-centre randomised clinical trial

Abstract Study question What is the effectiveness of swim-up (SU) and density gradients (DG) for sperm preparation in infertile couples undergoing intrauterine insemination (IUI)? Summary answer In couples undergoing IUI, there is no significant difference in the treatment outcomes between SU and DG. What is known already Swim-up (SU) and density gradients (DG) are the most two commonly used techniques for sperm preparation in intrauterine insemination (IUI). The latest Cochrane review concluded that there is uncertainty about the effectiveness of SU versus DG and no studies reported on live birth. Study design, size, duration This open-label, multi-centre randomised clinical trial was conducted on 912 infertile couples at a tertiary IVF center, Ho Chi Minh City, Vietnam from August 2020 to June 2023. Couples were randomly asigned to two groups, SU and DG using computerized randomisation on the day of IUI with variable block size of 2, 4 or 6. Primary outcome was live birth rate. Participants/materials, setting, methods Infertile couples were eligible if aged ≥18, indicated for IUI with ≤2 IUI prior attempts and husbands’ sperm concentration ≥5x106/ml, progressive motility (PR) ≥32% and total PR sperm count &amp;gt;5x106 before sperm preparation (WHO 2010 criteria). Couples using frozen semen, or high viscosity semen were not eligible. Women had ovulation induction using hMG. Sperm preparation was performed within one hour after ejaculation. IUI was performed once at 36 to 40 hours after hCG trigger. Main results and the role of chance Between August 2020 and June 2023, 912 couples were randomly assigned to SU (n = 456) or DG (n = 456). Baseline characteristics were comparable between the two groups (mean female age of 31.3 ± 0.4 years; total sperm count: 86.2 [52.9;137.6] million; sperm motility: 44.8 ± 8.2%). Live birth occurred in 55 (12.1%) couples in the SU group and 71 (15.7%) couples in the DG group (RR 0.77; 95% CI 0.56-1.07). Ongoing pregnancy and multiple pregnancy were also comparable between the two groups. There were no statistically significant differences between the SU and DG groups with respect to the occurrence of pregnancy complications, obstetric and perinatal complications, preterm delivery, birth weight and neonatal complications. Pre-planned subgroup analysis showed that the live birth rate was not materially affected by the IUI indications, total motile sperm after preparation (&amp;lt;1, ≥1 and &amp;lt;5, ≥5 and &amp;lt;10 and ≥10 million). Limitations, reasons for caution The main limitation of the study was its open-label design, due to the nature of the interventions. Wider implications of the findings Our study suggests that in couples undergoing IUI with progressive motility (PR) ≥32% and total PR sperm count &amp;gt;5x106 before sperm preparation, SU or DG can both be used for sperm preparation, with a preference for density gradients. Trial registration number NCT04477356

P-363 Reproductive toxicity of new immunobiologic treatments for male patients with melanoma

Abstract Study question Do immunotherapy and targeted therapy for melanoma affect male fertility? Summary answer Immunotherapy and targeted therapy for melanoma affect sperm quality. However, the detrimental effects are only transient. Sperm quality recovers once the treatment is discontinued. What is known already Immune checkpoint inhibitors, such as anti-BRAF and anti-MEK inhibitor, are currently used in melanoma patients to modulate the immune system and promote its anti-cancer action. Since these agents do not have an anti-mitotic action, they should not cause damage to spermatogenesis. However, no specific studies on human fertility have been conducted before drug registration. In fact, based only on pre-clinical studies, fertility-lowering effect of the BRAF/MEK inhibitors cannot be excluded. Potential endocrine autoimmune-related adverse events which could alter the hypothalamic-pituitary-gonadal axis such as hypophysitis, could impair spermatogenesis. These potential harmful effects should not be overlooked when counselling patients. Study design, size, duration Male patients scheduled for immunotherapies for melanoma were prospectively recruited at National Cancer Institute in Milan, Italy, between March 2020 and April 2023. Patients were counselled by a reproductive medicine specialist and offered to cryopreserve semen before immunotherapies. A semen analysis was also proposed 6-8 months after the initiation and 6-8 months after the end of immunotherapies. The primary outcome was to compare semen variables before, during and after immunotherapies. Participants/materials, setting, methods All patients underwent therapy with Nivolumab or Dabrafenib for 12 months.Semen samples were all cryopreserved and analysed by a single biologist at the Infertility Unit of Policlinico in Milan. Data are reported as median [Interquartile range-IQR] or number (percentage). Main results and the role of chance Twenty-three men operated on for melanoma and scheduled for immunotherapy were recruited. The age of patients at cryopreservation was 33 years [29–39]. An average of 12 [10-12] sperm paillettes were cryopreserved per patient. Total sperm count and total progressive motile sperm count were 126 millions [75–246] and 90 millions [38–154], respectively. Three patients were lost to follow-up and two patients died after cryopreservation. Twelve patients (52%) returned for a semen analysis during oncological treatment. After a median time of 9 months [8-12] of immunotherapy, a reduction in all semen variables was noted, particularly in total sperm count (77 million [49-189], p = 0.05 compared to basal condition) and the total count of progressive spermatozoa (34 million [23-77], p = 0.02 compared to basal condition). No differences were observed in morphology. Ten (43%) patients returned after the end of immunotherapies. One patient died before completing the follow-up. A recovery of all sperm variables compared to the cryopreserved sample was observed. Total sperm count and total motile sperm count were 190 millions [107–320] and 79 millions [34–140], respectively (p=ns). Two patients thawed their cryopreserved semen for an IVF cycle during immunotherapy. No pregnancy was achieved. Limitations, reasons for caution Our study evaluates male fertility only in terms of semen quality. Alterations of the hypothalamic-pituitary-gonad axis, such as the increase of autoimmune phenomena, have not been studied. Furthermore, the small sample size of our study is a limit. Wider implications of the findings Our study evaluates male fertility only in terms of semen quality. Alterations of the hypothalamic-pituitary-gonad axis, such as the increase of autoimmune phenomena, have not been studied. Furthermore, the small sample size of our study is a limit. Trial registration number not applicable

P-040 The characteristic of human seminal plasma extracellular vesicle (SPEVs) profiles with different sperm features in a large cohort of 309 Taiwanese men

Abstract Study question To investigate human seminal plasma extracellular vesicle profiles in different sperm characteristics and their potential clinical application. Summary answer Normozoospermic semen has significantly higher SPEV concentration than asthenozoospermic or oligozoospermic samples. Moreover, SPEV concentration was highly correlated with total motile sperm count. What is known already Seminal plasma extracellular vesicles (SPEVs) are cell-derived membrane structures that carry proteins, nucleic acid, and lipids, which can modify the constitution and biological function of sperm. However, the characteristics of SPEVs in different sperm features remained unclear. Study design, size, duration From January 2021 to September 2023, 309 men who visited the NCKUH andrology clinic agreed to collect semen samples for this study, which the Institutional Review Board has approved. Participants/materials, setting, methods Semen parameters were measured by the X1 CASA system (Bonraybio, Taiwan). SPEVs were isolated by size exclusion chromatography column and ExoQuick™. Next, the SPEVs’ concentration and size distribution were performed with nanoparticle tracking analysis. The Kruskal-Wallis test calculated comparison in multiple groups, and Dunn’s multiple comparison tests accessed group-group comparison. The Pearson correlation coefficient r of exosome concentration with individual sperm parameters was calculated. A p-value less than 0.05 was set as statistical significance. Main results and the role of chance Among them, 59 semen samples were presented with normozoospermia, 36 were oligozoospermia, 73 were asthenozoospermia, and 141 samples had no sperm in semen. Among these 141 azoospermic men, 89 men are characterized by non-obstructive azoospermia (NOA), while the other 52 men had a vasectomy. Our data showed that normozoospermic semen has significantly higher SPEV concentration than asthenozoospermic or oligozoospermic semen samples (26 x109 [IQR 17.25 x109-56.25 x109] versus 12 x109 [IQR 5.6 x109-19.5 x109] versus 15.5 x109 [IQR 8.575 x109-26.75 x109], p = 0.0001). Nevertheless, azoospermic semen has the lowest SPEV concentrations. The concentration of SPEVs from infertile men diagnosed with NOA is slightly lower than vasectomized men (3.9 x109 [IQR 1.875 x109-7.675 x109] versus 5.4 x109 [IQR 2.9 x109-8 x109], p = 0.0001). The Pearson r coefficient of SPEV concentration with sperm concentration, progressive motility, and total motile sperm count was 0.238, 0.303, and 0.451, respectively. (P &amp;lt; 0.0001) Limitations, reasons for caution Because our data were collected from Taiwanese men, it is not yet known whether SPEV profiles differ among races. Wider implications of the findings Human SPEV concentration and its bio-content may be a potential non-invasive biomarker to predict the male reproductive outcome. Trial registration number not applicable