Effect of Adding Human Umbilical Cord Mesenchymal Stem Cells-Derived Secretome on Sperm Quality Improvement by Swim-Up Method.

Study question Following 6thWHO (2021), we analysed the correlation between DNA fragmentation index (DFI), Human papillomavirus (HPV), and seminal parameters, highlighting slow and rapid progressive motility alterations. Summary answer DFI rates and seminal parameters correlated with rapid, slow, and progressive motility. However, HPV-positivity caused the loss of association between DFI and slow progressive motility. What is known already HPV detection in semen samples has long opened an investigation into its influence on male infertility. Some studies indicate that HPV can affect sperm quality and DFI, while others have failed to find any correlation. With reference to 2010 WHO guidelines, our latest work highlighted how HPV positivity significantly impairs progressive motility, morphology, and immotile sperm rate. Since the latest 2021 WHO guidelines included the evaluation of slow and rapid progressive motility and DFI, we analysed if these new parameters and the other conventional parameters could be altered by HPV infection. Study design, size, duration From August 2021 to December 2022, 121 semen samples were collected from male partners of HPV-positive women attending in vitro fertilization (IVF). Every specimen underwent DFI evaluation, analysis of seminal parameters, and HPV test. Participants/materials, setting, methods Seminal samples were collected by masturbation after 3-5 days of sexual abstinence. The inclusion criteria were as follows: no other sexually transmitted infections, no genetic diseases, and no inflammatory disorders. Sperm concentration, morphology, non-progressive and immotile sperms, and both slow and rapid progressive motility were evaluated according to WHO 2021 guidelines. DFI analysis was assessed by sperm chromatin dispersion test (SCD), while HPV-DNA detection was performed using InnoLipa HPV Genotyping Extra II (Fujirebio, Tokyo, Japan). Main results and the role of chance Of the 121 semen samples tested, 60 (49.6%) were HPV-positive and 61 (50.4%) were HPV-negative. DFI rates showed a significant negative correlation with rapid progressive motility in both groups and a positive correlation with slow progressive motility in the HPV-negative group. Conversely, the significance of the correlation between DFI and slow progressive motility was completely lost in HPV-positive patients. Sperm concentration, normal forms and immotile spermatozoa percentages were correlated with both motility parameters in the HPV-negative group. Similar results were observed in HPV-positive samples, except for the normal form rate, which was not associated with slow progressive motility. In addition, the same samples displayed a negative correlation between non-progressive motility and rapid progressive motility, absent in HPV-negative samples. Significant associations were found also for the derived parameter of progressive motility, which was correlated with DFI, sperm concentration, immotile sperm, and normal forms rate in both groups. The results suggest how high DFI rates, in the presence or absence of HPV infection, could affect reproductive health through a consistent impairment of spermatozoa motility. In particular, the distinction of slow and rapid progressive motility by WHO 2021 allows a deeper understanding of the possible correlations between DFI, semen parameters and HPV infection. Limitations, reasons for caution This is a preliminary study characterized by a small number of samples. Therefore, confirmation of these findings requires the enlargement of the patient cohort, which is already taking place. Wider implications of the findings Our results highlight how the introduction of the new WHO 2021 evaluation criteria, i.e. DFI, and slow and rapid progressive motility, provides additional information about sperm quality and the impact of HPV infection on it. Trial registration number Not applicable

To determine whether semen quality in Slovenians has changed over 14 years (1983-96), we analysed retrospectively the semen of 2343 healthy men with a normal spermiogram, who were partners of women with tubal infertility included in the IVF-ET programme. Age at semen collection, duration of sexual abstinence, semen volume, sperm concentration, total sperm count, percentage of spermatozoa with progressive motility, and normal morphology were determined. Multiple regression analysis was used to assess the changes in sperm characteristics according to the year of semen collection, year of the man's birth and the duration of sexual abstinence. Semen volume, sperm concentration, sperm count and total sperm motility did not change between 1983 and 1996, whereas between 1988 and 1996 rapid progressive sperm motility decreased by 0.95% per year (p < 0.0001). Semen volume, sperm concentration, and sperm count increased with duration of sexual abstinence. After adjustment for the year of semen collection and duration of sexual abstinence, multiple regression analysis showed that sperm concentration decreased by 0.67% per each successive year of birth (p = 0.03). Thus the sperm concentration decreased from 87.6 x 10(6)/mL in men born in the 1940s to 77.3 x 10(6)/mL in those born between 1956 and 1960. After 1960, sperm concentration was found to increase. In 2343 healthy men, no decline in semen quality, except in rapid progressive motility, was observed in the study period. Lower sperm concentration was found among men born between 1950 and 1960. This could be related to worse socio-economic status, stress or negative environmental factors in this time period.

Processing of semen can result in increased sperm DNA fragmentation

The objective of this study was to evaluate sperm quality of Duroc boars during sexual maturation and examine whether these parameters were related to metabolites in semen. Ejaculates were collected longitudinally from 28 Duroc boars at approximately 7 months of age (Age 1), 6 weeks later (Age 2) and 12 weeks later (Age 3). Motility characteristics, acrosome integrity, and viability were assessed on collection day and after five days storage at 18°C using computer-assisted sperm analysis and flow cytometry, while DNA fragmentation was measured on day five. Amino acid and amine concentrations in semen were measured by liquid chromatography-mass spectrometry and correlated with sperm quality parameters. Motility, rapid progressive motility, and acrosome intact live sperm increased significantly (adj P < 0.05) with boar maturation. Mature boars (Age 2 and Age 3) maintained superior sperm quality compared to younger boars after storage. The DNA fragmentation index was low in all samples but declined significantly from 1 % at Age 1 to 0.64 % at Age 3 (adj P < 0.05). Correlation analysis showed significant relationships between sperm quality and specific metabolites. Cystine, glutamate, aspartate, choline and taurine were inversely correlated with progressive and rapid progressive motility, while showing positive correlation with rapid non-progressive motility. These findings demonstrate that sperm quality continues improving beyond initial reproductive ability, especially between Age 1 and Age 3, with improvements in motility, viability, chromatin stability, and storage resilience. The observed relationships between metabolites and sperm quality parameters provide insights into biochemical mechanisms underlying sperm functionality during sexual maturation.

Nurture vs nature: how can we optimize sperm quality?

Science linking environmental contaminant exposures with fertility and reproductive health impacts in the adult male

A live motile sperm sorting device (LensHooke® CA0) developed to prevent the deleterious effects of centrifugation was evaluated comparatively with conventional density-gradient centrifugation (DGC) and microfluidic-based device (Zymot) in sperm selection. Semen samples from 239 men were collected. CA0 under different incubation intervals (5, 10, 30, and 60min) and temperatures (20, 25, and 37℃) was conducted. The sperm quality in CA0-, DGC-, and Zymot-processed samples was then comparatively evaluated. Semen parameters included concentration, motility, morphology, motion kinematics, DNA fragmentation index (DFI), and the rate of acrosome-reacted sperm (AR). Total motility and motile sperm concentration increased in a time- and temperature-dependent manner and the total motility peaked for 30min at 37℃. In paired analysis, CA0 showed significantly higher total motility (94.0%), progressive motility (90.8%), rapid progressive motility (83.6%), normal morphology (10.3%), and lower DFI (2.4%) and AR (4.7%) than the other two methods in normozoospermic samples (all p < 0.05). For non-normozoospermic samples, CA0 had significantly better results than the other two methods (total motility 89.2%, progressive motility 80.4%, rapid progressive motility 74.2%, normal morphology 8.5%, DFI 4.0%, and AR 4.0%; all p < 0.05). CA0 yielded spermatozoa with enhanced sperm fertilization potentials; DFI was minimized in samples processed by CA0. CA0 was effective for both normal and abnormal semen samples due to its consistent selection efficiency.

Infertility affects 186 million people worldwide, with male factors contributing to 50% of infertility cases. Semen analysis is a key for diagnosing male factor infertility, but sperm parameters can be influenced by ejaculatory abstinence (EA) duration. Shortening or prolonging EA can impact on semen quality and assisted reproductive technology (ART) outcomes, but the optimal EA duration remains unclear, particularly for infertility patients. This study conducts a comprehensive meta-analysis to explore the impact of varying abstinence durations on semen quality and fertility outcomes. Three English database (PubMed, Embase, and Cochrane Central Register of Controlled Trials) as well as four Chinese database (China National Knowledge Infrastructure, Chinese Scientific Journals database, WanFang database, and Chinese Biomedical Literature database) were searched from 2000 to August 2023. The classical meta-analysis and "one-stage" dose-response meta-analysis were conducted to compare the associations of different abstinence durations (short-term abstinence vs. long-term abstinence) on semen quality in healthy adult and different type of infertile patients. There were 85 eligible studies were finally included. The meta-analysis of volume (mean difference [MD]=-0.95mL, 95% confidence interval [CI]: -1.16 to -0.74mL), total sperm count (TSC) (MD=-102.45×106, 95% CI: -117.98×106 to -86.91×106), sperm concentration (SC) (MD=-11.88×106/mL, 95% CI: -18.96×106/mL to -4.80×106/mL), DNA fragmentation index (DFI) (MD=-2.37%, 95% CI: -4.73% to -0.01%) in healthy men showed a significant decrease with different abstinence durations (short-term abstinence vs. long-term abstinence). The meta-analysis of infertile men showed significant decrease in volume in various subgroups (MD range: -0.73 to -1.17mL) with P<0.01; TSC (MD=-61.93×106, 95% CI: -88.84×106 to -35.01×106), SC (MD=-5.39×106/mL, 95% CI: -9.97×106 to -0.81×106/mL), DFI (MD=-5.63%, 95% CI: -10.19% to -1.06%) in unexplained infertility subgroup; significant increase in viability (MD=6.14%, 95% CI: 3.61% to 8.68%) in the unexplained infertility subgroup. The dose-response meta-analysis showed that TSC in oligozoospermia showed a nonlinear increase (coefficient from 3.38 to -5.76, P from 0.02 to 0.22) and the truncation point was around the 4th to 5th abstinence day. The percentage of progressive motile sperm (PR) in asthenozoospermia showed a significant decrease (coefficient=-2.39, 95% CI: -4.28 to -0.50). For fertility outcomes of different ARTs, only the clinical pregnancy rate (CPR) in the intrauterine insemination (IUI) subgroup showed a significant decrease around the 3rd day (coefficient=0.85, 95% CI: 0.75 to 0.97). Short-term abstinence may be associated with limited improvements in semen quality in healthy men but could be more beneficial for infertile men, especially within the first 4 days of abstinence. Caution is urged in making definitive conclusions about the causal relationship between abstinence time and semen quality changes due to potential confounding and interactions.

Study question Can ORP serve as a reliable biomarker for male infertility assessment and should it be integrated into routine testing? Summary answer Since ORP does not correlate with DNA fragmentation, both parameters should be routinely assessed in male infertility evaluations for a comprehensive understanding of semen quality. What is known already Oxidative stress plays a crucial role in male infertility by impairing sperm function and increasing DNA fragmentation. ORP is an emerging marker to evaluate oxidative stress levels in semen. Elevated ORP has been associated with poor semen quality, including reduced motility and concentration. However, its relationship with SDF remains unclear, and studies assessing its predictive value for male infertility are limited. Understanding the association between ORP, sperm concentration, and SDF may help refine diagnostic protocols and improve patient management. Study design, size, duration This retrospective cohort study analyzed semen samples from 900 men undergoing fertility evaluation over two years. Participants/materials, setting, methods This study included men (mean age: 38 years) undergoing fertility evaluation. Semen samples were collected and analyzed following WHO guidelines to ensure standardized assessment of oxidative stress and sperm quality. Sperm DNA fragmentation (SDF) was measured using the TUNEL assay on the BD FACSLyric flow cytometer, while oxidation-reduction potential (ORP) was assessed with the MiOXSYS analyzer. Semen parameters, including concentration, total sperm count, pH, and abstinence duration, were also recorded. Main results and the role of chance Semen analysis of 900 men undergoing fertility evaluation showed a mean sperm concentration of 57.2 × 106 sperm/mL and a total sperm count of 147.9 × 106 sperm/sample. The mean semen pH was 8.2, and the average abstinence period was 2.9 days. ORP levels averaged 1.7 mV/106 sperm/mL, with values &amp;gt;1.34 mV/106 sperm/mL indicating high oxidative stress. The mean SDF was 19.4%, categorized as low (≤16.9%), moderate (16.9–30%), or high (&amp;gt;30%). The distribution of ORP and SDF categories among patients was: low ORP &amp; low SDF (38.4%), low ORP &amp; moderate SDF (22.6%), low ORP &amp; high SDF (11.8%), high ORP &amp; low SDF (13.8%), high ORP &amp; moderate SDF (7.3%), and high ORP &amp; high SDF (6.1%). A weak, non-significant correlation was found between ORP and SDF (Spearman r = 0.046, 95% CI: -0.02 to 0.11, P = 0.46). However, a significant negative correlation was observed between ORP and sperm concentration (Spearman r = -0.569, 95% CI: -0.61 to -0.52, P &amp;lt; 0.001). Limitations, reasons for caution Potential methodological variations in ORP and SDF measurement techniques may impact results. Standardization across different assay platforms and laboratory conditions is necessary to ensure consistency and reproducibility. Wider implications of the findings ORP is a valuable complementary marker for semen quality, helping identify infertile patients with normal DNA fragmentation. Since oxidative stress affects fertility independently of DFI, evaluating both markers in male infertility assessments could improve diagnostic accuracy and guide personalized treatment strategies, ultimately enhancing reproductive outcomes for affected individuals. Trial registration number No

Due to the limitations of conventional semen analysis in predicting a man's fertility potential, sperm DNA fragmentation was recently introduced as a novel marker of sperm quality. This prospective study was undertaken to investigate the associations between conventional seminal parameters and DNA fragmentation in Greek men. A total of 669 subject data were evaluated in two groups, normozoospermic (n = 184) and non-normozoospermic (n = 485), according to the WHO 2010 (WHO Laboratory Manual for the Examination and Processing of Human Semen, 5th edn. World Health Organization), reference limits. For all the subjects, semen volume, sperm concentration, total count, rapid and total progressive motility and morphology were recorded following the WHO 2010 methods and DNA fragmentation was assessed by the sperm chromatin dispersion assay. An inverse correlation was established between DNA fragmentation and all conventional seminal parameters except semen volume in men with seminal profiles below the reference limits, with statistical significance for rapid and total progressive motility. Normozoospermic men exhibited lower levels of DNA fragmentation than their non-normozoospermic counterparts, even though the values were not always below 30%. DNA fragmentation testing and traditional semen analysis should therefore be considered as complementary diagnostic tools in a comprehensive evaluation of male infertility.

Study question How does convolutional neural network (CNN)-predicted sperm motility correlate with manual assessment according to the WHO guidelines. Summary answer CNN predicts sperm motility comparable to reference laboratories in the ESHRE-SIGA External Quality Assessment Programme for Semen Analysis. What is known already Manual sperm motility assessment according to WHO guidelines is regarded as the gold standard. To obtain reliable and reproducible results, comprehensive training is essential as well as running internal and external quality control. Prediction based on artificial intelligence can potentially transfer human-level performance into models that perform the task faster and can avoid human assessor variations. CNNs have been groundbreaking in image processing. To develop AI models with high predictive power, the data set used should be of high quality and sperm motility assessment based on WHO guidelines. Study design, size, duration Videos of 65 fresh semen samples obtained from the ESHRE-SIGA External Quality Assessment Programme for Semen Analysis (from the period 2006–2018) were used in the development of the model. One video was captured for each semen sample. Sperm motility data was obtained from manual assessment of the videos according to WHO criteria by reference laboratories in the programme. Rapid progressive motility was also included. Ten-fold cross-validation was used to compensate for the relatively small dataset. Participants/materials, setting, methods The mean values of the reference laboratories were used. Sparse optical flow of the sperm videos was generated from each second of each video and fed into a ResNet50 convolutional neural network. For training, Adam was used to optimize the weights and mean squared error (MSE) to measure loss. For baseline, ZeroR (pseudo regression) was performed. Results are reported as MAE. For correlation analysis, Pearson’s r was used. Main results and the role of chance Predicting sperm motility based on the optical flow generated from the videos, achieved an average MAE of 0.05 across progressive (0.06), non-progressive (0.04) and immotile sperm (0.05). The ZeroR baseline was 0.09, indicating that the method is able to capture the movement of the spermatozoa and predict motility with low error. Pearson’s correlation between manually and AI-predicted motility showed r of 0.88, p &amp;lt; 0.001 for progressive, 0.59, p &amp;lt; 0.001 for non-progressive and 0.89, p &amp;lt; 0.001 for immotile sperm. When predicting rapid progressive motility, the average MAE was 0.07 across rapid progressive (0.11), slow progressive (0.09), non-progressive (0.04) and immotile sperm (0.05). Pearson’s correlation analysis between manually and AI-predicted motility showed r of 0.67, p &amp;lt; 0.001 for rapid progressive, 0.41, p &amp;lt; 0.001 for slow progressive, 0.51, p &amp;lt; 0.001 for non-progressive and 0.88, p &amp;lt; 0.001 for immotile sperm. The results show that differentiating between rapid progressive and slow progressive motility is difficult, but the model is still able to do this better than the ZeroR baseline, which was 0.15 for rapid progressive and 0.11 for slow progressive. This is interesting since rapid progressive motility has been regarded challenging to assess. The next step would be to compare the results of the algorithm to the human performance. Limitations, reasons for caution The sample size is small. The model is based on videos of high quality, and the performance may not transfer well to videos of lower quality. The performance for rapid progressive motility, which may have an important clinical value, has to be improved. Wider implications of the findings: This CNN model has a potential to assess sperm motility according to WHO guidelines for progressive motility and immotility. The error values for the automatic predictions are low, and the model shows a good performance taking into account that only videos were used to perform the prediction. Trial registration number Not applicable

Study question Do sperm quality parameters differ between “high” and “low” cardiorespiratory fitness (CRF) and muscular strength levels in healthy young men? Summary answer Self-reported CRF was positively associated with sperm count and concentration, while muscular strength (objectively measured and self-reported) did not correlate with seminal parameters. What is known already Physical activity (i.e., any body movement that results in energy expenditure) levels are positively associated with sperm quality parameters in healthy men. In this context, physical fitness, which is modifiable by physical activity, is a powerful marker of health that integrates several body functions such as hematocirculatory, skeletomuscular, phsychoneurological and endocrine-metabolic. In particular, the main components of physical fitness (i.e., CRF and muscular strength) have been negatively related to the risk of all-cause mortality in any age population. However, the association of CRF and muscular strength with sperm quality parameters in healthy young men is unknown. Study design, size, duration This cross-sectional study included 253 healthy men (age= 22.63 ± 5.04 years, BMI= 22.84 ± 1.33). Semen samples and physical fitness assessment were performed between March 2019 and July 2021 at the sperm biobank (Ceifer Biobank – NextClinics). The study was approved by the Ethics Committee of Investigación Biomédica de Andalucía. Participants/materials, setting, methods CRF was self-reported using the The International FItness Scale (IFIS). Muscular strength was objectively assessed using a handgrip dynamometer and self-reported with IFIS. Semen samples were analysed by clinical technicians to determine sperm count, sperm concentration and progressive motility. Analysis of covariance (ANCOVA) was performed to obtain adjusted mean differences on sperm quality parameters between high and low CRF and muscular strength groups after including age and body mass index as confounders. Main results and the role of chance Men with higher CRF presented increased sperm count and concentration compared to men with lower CRF values (adjusted mean differences 43.581 x 106 [2.630, 84.532; 95% confidence interval] and 10.131 x 106/ml [0.799, 19.463; 95% confidence interval]; both p &amp;lt; 0.05). Muscular strength (objectively measured and self-reported) was not associated with any semen quality parameter (all p &amp;gt; 0.05) when including age and body mass index as covariates. Limitations, reasons for caution This study reported a positive association between self-reported CRF using IFIS questionnaire and sperm count and concentration in healthy young men. Although the validity of the self-reported CRF using IFIS has been widely proved in healthy young adults, objectively measured CRF is needed to further confirm our findings. Wider implications of the findings Infertility has increased in the last decades. Therefore, more research is needed to identify health indicators that affect sperm quality. Our results suggest that self-reported CRF could contribute to the identification of young men with poor seminal quality and a higher risk of infertility later in life. Trial registration number NA

Study question Can centrifugation-free sperm separation device be used to simplify and standardize the selection of high-quality spermatozoa? Summary answer Centrifugation-free CA0 sperm separation device minimizes inter- and intra-operator variability and yields spermatozoa with comparable fertilizing properties in a variety of semen conditions. What is known already Centrifugation-based sperm separation methods have been used in assisted reproductive technology (ART) for many decades. However, the conventional methods are criticized for harmful effects due to centrifugation. To overcome the disadvantage, more noninvasive technologies have been developed and attempted to improve the sperm separation process, e.g., the formation of capillary bridge to select motile spermatozoa, migration-sedimentation technique to sort out functional spermatozoa, microfluidic sorting chip to isolate healthy sperm. While these methods provide alternatives for noninvasive sperm separation, the limitations such as inconsistency of semen quality improvement and lack of standardized procedure remain to be resolved. Study design, size, duration A randomized controlled trial of 76 men who sought ART treatment (Lee Women’s Hospital, Taichung, Taiwan) from June to October 2022 was carried out. Seventy-six neat semen specimens were categorized into 27 normozoospermic specimens, and 49 non-normozoospermic samples (semen quality below any of the WHO 5th Edition lower reference values: concentration &amp;lt;15 million/mL, total motility &amp;lt;40%, or normal morphology &amp;lt;4%). Participants/materials, setting, methods Neat semen samples were separated for three replicates (replicate 1-3) using LensHooke® CA0 sperm separation device (Bonraybio, Taichung, Taiwan). Three operators (operators 1–3) performed CA0 procedures on each sample to test inter-technician variability. Interclass-correlation coefficient (ICC) between replicates as well as operators were evaluated. Pre-selection and post-selection semen quality were evaluated. The parameters included total motility, progressive motility, rapid progressive motility, morphology, DNA fragmentation index (DFI), and acrosome reaction rate (AR). Main results and the role of chance CA0 selects self-propelling spermatozoa within a microenvironment created by a microporous filter membrane. The procedure involves three pipetting steps: loading semen sample, adding sperm washing medium, and recovering the processed sample. Following the standard procedure, CA0 resulted in a low intra- and inter-operator variability and ICC values between replicates and different operators were all greater than 0.9, indicating an excellent reproducibility of CA0. In addition, significant higher levels of motility and normal morphology were observed in post-selection specimens either of normozoospermic or non-normozoospermic samples (pre-selection vs. post-selection, all p &amp;lt; 0.0001). In paired analysis of the advanced semen parameter, our study showed noteworthy results that CA0 significantly improved DFI from 18.2% to 2.6% for normozoospermic samples; such reduction was also found in non-normozoospermic sample processing, from 13.4% to 4.2% (both, p &amp;lt; 0.0001). The levels of AR were significantly reduced in normozoospermic samples (from 14.4% to 5.4%) and non-normozoospermic samples as well (from 12.8% to 5.0%) (both groups p &amp;lt; 0.0001). In conclusion, CA0 provides an efficient, noninvasive, standardized, and reproducible sperm separation model that CA0 diminishes the variations and ensures sperm quality. Limitations, reasons for caution The presented study was a pilot trial examining the sperm quality improvement. Follow-up analysis on IUI/IVF outcomes associated with the improvement in semen quality utilizing CA0 will be assessed in future studies. Wider implications of the findings CA0 provides multifaceted benefits covering consistent clinical outcomes, simplified and standardized procedure, user-friendliness, and cost reduction. We believe CA0 not only allows noninvasive sperm separation of clinically usable but also gives the possibility of standardization on sperm separation procedure. Trial registration number CS2-22039

Background:Obesity, Vitamin D (VD) deficiency, and infertility are important ubiquitous issue; however, the association of obesity and serum VD levels with abnormal sperm is unclear and inconclusive. The current study investigated the correlation of obesity and serum VD levels with sperm DNA integrity and sperm parameters in normozoospermia men.Materials and Methods:Semen and blood samples from 64 men were divided into two groups: obese and nonobese men based on body mass index (BMI). Sperm motility and viability were determined by computer-aided sperm analysis and eosin-nigrosin staining. DNA fragmentation, determined by sperm chromatin dispersion method. VD concentrations were assessed by the Elisa technique.Results:Serum concentration of VD levels in the obese group was significantly lower than nonobese men (P < 0.05). Sperm motility was significantly reduced in the obese group in comparison to nonobese (P < 0.05). Rapid progressive motility was statistically lower in obese men compared with the nonobese group (P < 0.05). Sperm count and morphology were not statistically significant in both groups. Sperm viability in the nonobese group was significantly decreased in comparison to obese group (P < 0.05). DNA integrity was significantly higher in the obese group as compared with nonobese (P < 0.01).Conclusion:VD deficiency in the obese group showed decreased sperm motility, increased DNA damage, and viability. Adverse consequences of obesity and the possible effect of BMI infertility treatment must be discussed with counseling couples interested in assisted reproductive techniques outcomes, especially in men without any unknown cause.

Male infertility is a complex issue influenced by multiple environmental and pathological factors. In this context, the impact of Human papillomavirus (HPV) infection on male fertility remains controversial. The introduction of new WHO 2021 evaluation criteria, included in the 6th ed. of Laboratory Manual for the examination and processing of human semen, i.e. DNA fragmentation index (DFI), slow and rapid progressive motility, could provide additional information about this correlation. 121 semen samples of male partners of HPV-positive women attending In Vitro Fertilization (IVF) were evaluated following WHO 2021 and HPV-DNA test. Comparing HPV-negative and positive samples for rapid and slow progressive motility showed significantly different results (p = 0.0018, p = 0.0004), contrary to what was observed for total progressive motility. Regarding sperm DFI, only high-risk HPV infections affected DNA integrity. In addition, the correlation between the different semen parameters revealed a significant correlation between midpiece morphological defects and rapid progressive motility in the HPV-positive group (rho = 0.43, p = 0.0006). In conclusion, WHO 2021 provides additional information regarding HPV’s impact on seminal parameters. The correlation between HPV positivity, midpiece defects and a higher rapid progressive motility opens new research perspectives that may help unravel the issues surrounding the role of HPV in compromising sperm quality.