Abstract Background Semen analysis remains the first‐line diagnostic test for male infertility, although it is affected by operator subjectivity. Computer‐assisted sperm analysis (CASA) is proposed to improve objectivity; however, its accuracy and clinical relevance remain uncertain. Objectives We evaluated 424 samples from men attending routine semen analysis and 179 samples from male partners of ICSI couples to compare manual assessment with a CASA system (Ceros II), and to explore whether CASA‐derived kinematic parameters, in particular hyperactivated motility (HA), predict ICSI outcomes. Materials and Methods Manual (according to WHO manual) and CASA assessments were compared using correlation analysis, Passing–Bablok regression, and Bland–Altman plots; kinematic parameters were evaluated in relation to ICSI outcomes. Results Significant correlations were found between CASA and manual assessment for rapid progressive motility and for sperm count for samples with less than 50 × 10 6 /mL spermatozoa, whereas agreement was lost above this value, unless dilution was performed. Among CASA kinematic parameters, HA measured in whole semen was significantly higher in cycles achieving clinical pregnancy and resulted an independent pregnancy predictor also after adjustment for female age and female factor, showing good discriminative performance. Discussion These findings suggest that CASA provides objective, reproducible assessment of sperm motility, and that hyperactivated motility may serve as a clinically relevant marker to improve prediction of ICSI outcomes. Conclusions CASA may provide useful support for routine semen analysis and identify clinically meaningful kinematic parameters. Trial Registration Not applicable

The management of species in human care, hereafter ex situ management, has been identified as a key tool in the effort to restore global biodiversity. However, sustaining reproduction and genetic diversity in an ex situ environment can be challenging. Assisted reproductive technologies (assisted reproduction) can help. In this study, we begin development of an assisted reproduction program for an ex situ population of gray ratsnake (Pantherophis spiloides), a federally 'threatened' species in Canada. Assisted reproduction application in squamates has had limited attention, despite the disproportionate representation of the taxa among threatened and endangered species worldwide. Hence, we hoped to provide a model for the development of assisted reproduction in squamates in Canada more broadly. As such, we focused on the essential first steps of gamete collection and short-term cold preservation. We collected sperm samples monthly from 17 individuals in the ex situ population from May to October. We assessed concentration, motility, rapid progressive motility, and membrane integrity (viability). We used generalized linear models to assess the impact of four common sperm extenders on refrigerated sperm quality over a period of 5 days. We found that TL HEPES with 10% fetal bovine sera maintained the highest quality sperm. Further, we discovered previously unknown seasonality in sperm viability, where the highest percentage of viable sperm was collected in October, rather than May-June, at the height of the species breeding season. Our work highlights that the timing of sperm collection and the type of sperm extender used both influence the preservation of sperm ex vivo.

This study investigates the effects of high-dose radioactive iodine therapy on gonadotropin and sex hormone levels, and on sperm parameters in male patients with differentiated thyroid carcinoma following thyroidectomy. Twenty-five male patients (aged 20-60 years) with differentiated thyroid carcinoma underwent thyroidectomy and iodine therapy. The therapeutic dose was 150 mCi of oral sodium iodide solution. Levels of gonadotropins, sex hormones, and anti-Müllerian hormone (AMH) were measured before and two weeks after radioiodine therapy (RT). Semen analysis included liquefaction, odor, color, viscosity, agglutination, and aggregation. The main parameters evaluated were semen volume, pH, sperm count, percentages of motile and progressively motile sperm, round cells, and sperm morphology. Sperm motility, including progressive, non-progressive, and immotile types, and DNA fragmentation were analyzed according to World Health Organization guidelines. The Wilcoxon signed-rank test was used with a significance level of p≤0.05. Follicle-stimulating hormone levels in patients' sera were significantly higher than pre-RIT measurements (p=0.002), whereas luteinizing hormone, dihydrotestosterone, dehydroepiandrosterone sulfate, testosterone, and AMH levels were not significantly different from pre-RT measurements. Total sperm count, volume, motility, and rapid progressive motility increased significantly compared to pre-radioiodine ablation measurements, while other parameters remained unchanged. Male patients who received 150 mCi of radioactive iodine showed no impairment in fertility. Long-term follow-up studies with larger sample sizes are crucial to investigate the physiological roles of gonadal hormones, sperm DNA fragmentation, and AMH in the testes after RIT.

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.

ABSTRACTPurposeMammalian sperm require a large amount of ATP to sustain motility. While glycolysis and oxidative phosphorylation (OXPHOS) are established ATP sources, they cannot fully explain motility maintenance under glucose‐deprived conditions. This study examined whether gluconeogenesis functions as a third metabolic pathway supporting sperm energy production.MethodsSperm from mice, humans, cattle, and horses were comparatively analyzed under glucose‐containing and glucose‐free conditions with inhibitors of OXPHOS (FCCP) and fructose‐1,6‐bisphosphatase 1 (FBPase1). Motility and ATP levels were measured, and FBPase1 expression was assessed by Western blotting and immunofluorescence.ResultsMouse sperm maintained motility without glucose, and inhibition of FBPase1 significantly reduced ATP and rapid progressive motility. FBPase1 was expressed in the midpiece and principal piece, indicating active gluconeogenesis. Similar patterns were observed in human and bovine sperm, suggesting a conserved mechanism among mammals.ConclusionGluconeogenesis serves as a third metabolic platform that complements glycolysis and OXPHOS, enabling efficient substrate use and sustaining sperm motility. This discovery provides new insight into sperm metabolic flexibility and may contribute to treatments for male infertility and improved livestock reproduction.

Suboptimal sperm quality often poses a challenge to successful fertilization. This study hypothesizes that secretome could enhance sperm quality and increase the likelihood of successful pregnancy. This research aims to assess the impact of secretome on various sperm quality parameters, including concentration, motility, and DNA fragmentation, while considering patient-related factors such as age, duration of abstinence, and body mass index (BMI). An analysis involving 45 patients enrolled in the pregnancy program at Halim Fertility Center, Stella Maris Women's and Children's Hospital, Indonesia, from April to September 2023 was conducted. Semen samples from these patients were subjected to the swim-up method and divided into two groups: Group A, which underwent swim-up without secretome, and Group B, which underwent swim-up with the addition of secretome. DNA fragmentation analysis was performed on the sperm swim-up results from both groups. The sperm analysis data obtained before swim-up (pretreatment) with those of Group A and Group B were compared. The demographic data revealed an average age of 37.67 ± 5.36 years, abstinence duration of 4.00 ± 1.15 days, and BMI of 28.20 ± 3.49 kg/m2 among the patients. No significant difference was observed in sperm concentration between pretreatment, Group A, and Group B (mil/mL) (36.2 ± 18.5; 36.3 ± 18.4; 36.6 ± 19.3). However, a significant difference was found in the rapid progressive motility of sperm across pretreatment, Group A, and Group B (%) (0.48 ± 1.32; 13.7 ± 8.3; 17 ± 8.3), as well as a significant reduction in DNA fragmentation in Group B compared to Group A (%) (3.48 vs. 4.39). The findings suggest that secretome enhances rapid progressive motility and reduces DNA fragmentation rates without affecting sperm concentration.

The objective of the study was to investigate the vanillic acid's (VA) protective effects, a phenolic compound, on the ram semen after freeze-thaw. Semen was obtained from Ramlıç rams and was diluted with control (0 μg/mL VA) and VA-supplemented Tris-based extenders at concentrations of 1, 10 and 50 μg/mL. The diluted semen was equilibrated for 2 h at +4°C, filled into 0.25 mL straws and frozen in liquid nitrogen vapour. It was then stored in a liquid nitrogen container at -196°C. For analysis, the samples were thawed at 37°C for 30 s in a water bath. There was no difference detected among total and progressive motility as well as velocity parameters (p > 0.05) except for rapid progressive motility (p < 0.05). The findings support the idea that VA has an outstanding effect on reducing DNA damage (p < 0.001). While there was no positive development with regard to total oxidant status (p > 0.05), VA enhanced the antioxidant defences of total antioxidant status (p < 0.05). VA administered at doses of 10 and 50 μg increased total antioxidant status (p < 0.01). Lipid peroxidation was not directly affected by VA application (p > 0.05); otherwise, 10 and 50 μg VA treatments showed a positive effect on viability (p < 0.001). Based on findings, it was concluded that although VA was put in the semen extender, it did not have an ameliorative potency on sperm motility and velocity properties except for specific sub-parameters such as rapid progressive motility. All applied doses reduced DNA damage, and 10 and 50 μg doses supported cellular viability.

Deciphering the impact of essential elements on spermatozoa quality and seminal protein integrity- an in vitro and in silico study.

The role of sperm morphology in male fertilization potential remains unclear. This study aimed to evaluate the incidence of specific sperm morphological abnormalities in a cohort of Turkish men attending a single-center infertility clinic and to assess their associations with semen quality and sperm functionality. A total of 2,923 men aged 17–57 years were included in this cross-sectional study. Semen parameters and specific morphological abnormalities in the head, neck-midpiece, tail, and cytoplasmic residue were analyzed according to WHO criteria. Participants were categorized into normal and low semen parameter groups based on sperm count, motility, and normal morphology. Associations between semen parameters and morphological abnormalities were assessed using Spearman correlation and binary logistic regression analyses. Head defects were the most prevalent abnormalities, followed by tail, neck-midpiece, and cytoplasmic residue. Normal sperm morphology and specific abnormalities were significantly associated with semen quality, with normal forms demonstrating the strongest predictive potential. Head defects were primarily associated with teratozoospermia, whereas neck-midpiece and tail defects were linked to motility impairments. Notably, round and tapered heads, bent necks, and coiled or short tails showed the strongest associations with progressive and rapid progressive motility, which are critical determinants of fertilization capacity. Our findings indicate that different categories of sperm abnormalities have distinct predictive potentials for semen quality disorders and suggest that specific defects may arise from disruptions in different stages of sperm morphogenesis.

Assisted reproductive technologies, such as in vitro fertilization, remain inefficient in camelids, largely due to gaps in understanding the molecular interactions that regulate sperm capacitation. Fertilization requires not only viable spermatozoa but also the precise modulation of capacitation by the peri-ovulatory microenvironment, including follicular fluid (FF) and oviductal fluid (OF). In this study, spermatozoa were incubated in Fert-TALP medium supplemented with FF or OF, and both functional outcomes and proteomic remodeling were assessed. Sperm treatments were evaluated in five independent biological replicates per individual (three individuals), with triplicate proteomics performed. FF (n = 20) was collected from pre-ovulatory follicles (7–9 mm) and OF (n = 10) from the corresponding ipsilateral oviducts, thereby reflecting the in vivo environment encountered by sperm in the female reproductive tract following mating. Incubation with FF enhanced progressive motility by 72%, rapid progressive motility by 169%, viability by 30%, and acrosome responsiveness by 30%, and was associated with a proteomic shift involving ~12% of proteins (p < 0.05). These included factors implicated in zona pellucida binding (LYPD4, PGK1, ANXA2, and TCP1 complex members) and galactose metabolism (MAOA, AKR1B1, GLA, and HK1). The enriched processes included glycolysis/gluconeogenesis, cytoskeletal reorganization, and protein maturation, all consistent with sperm capacitation. By contrast, sperm incubated with OF showed an underrepresentation of capacitation-related pathways, including the proteasome complex, sperm fibrous sheath, and TCA cycle. Moreover, the OF proteome (r = 2) revealed decapacitation-associated factors such as PEBP1 and PAFAH1B3, which likely stabilize membranes and delay premature capacitation. Together, these findings demonstrate complementary yet contrasting roles of FF and OF in modulating sperm physiology: FF acting as a capacitating medium, and OF providing a stabilizing environment. This study presents the first partial proteome of capacitated alpaca sperm together with matched reproductive fluids, providing mechanistic insights with direct implications for improving assisted reproduction in camelids.

F4-Neuroprostanes (F4-NeuroPs), oxidative metabolites of docosahexaenoic acid, act as bioactive lipid mediators enhancing sperm motility and induce capacitation-like changes in vitro. Their biological action is proposed to involve sperm ion channels, in particular ryanodine receptors (RyRs), which regulate intracellular calcium homeostasis. We evaluated the effects of dantrolene, a RyR inhibitor, on motility and vitality of a selected spermatozoa at different concentrations (10, 30, 50, 100 μM). Then sperm motility, acrosome integrity, and RyR localization following co-incubation with dantrolene (D50 or D100 μM) and 4-/10-F4t-NeuroPs (7 ng) were investigated. Acrosomal status was assessed using Pisum sativum agglutinin (PSA) staining and RyR localization by immunofluorescence. D50 was identified as the minimum effective dose to induce significant reductions in sperm motility. F4-NeuroPs significantly increased rapid progressive motility versus controls. Co-incubation with F4-NeuroPs + D50 reduced rapid motility and increased in situ and circular movement. The acrosome staining appeared altered or absent to different percentages, and RyR localization was also seen in the midpiece. These findings suggested that F4-NeuroPs enhance sperm motility via RyR-mediated pathways, as confirmed by dantrolene inhibition. Accordingly, our results underscore the physiological relevance of RyRs in sperm function and suggest new insights into lipid-based mechanisms regulating sperm motility.

Sperm motility is a key indicator of male fertility. Decreased motility, or asthenozoospermia, highlights the need for understanding male fertility challenges. This study was designed to evaluate the effects of in vitro use of follicle-stimulating hormone (FSH) on various sperm parameters, sperm DNA integrity, and mitochondrial membrane potential. Semen samples were obtained from 20 asthenozoospermic men. Each sample was divided into control and case groups. In the case group, samples were incubated with FSH at 30mIU/mL for 1h, while in the control group, the samples were incubated for 1h without treatment. Sperm parameters, DNA fragmentation, and mitochondrial membrane potential were assessed in the two groups based on WHO 2021 criteria. Progressive motility and rapid progressive motility were higher in the case group compared to the control. However, FSH did not have a significant effect on morphology, viability, DNA fragmentation, or mitochondrial membrane potential in either group. In conclusion, FSH effectively enhanced sperm motility without compromising sperm DNA integrity or mitochondrial membrane potential (MMP). Therefore, FSH can be recommended as a safe and effective option for sperm selection in patients with asthenozoospermia.

The current hypothesis suggests that translation occurs in capacitated spermatozoa through mitochondrial ribosomes. Mitochondrial translation has several particularities, which rise some questions about how mitochondrial ribosomes can ensure sperm translation activity. Here, we aimed to elucidate if cytoplasmic translation occurs in mammalian spermatozoa. A bioinformatic workflow was performed to identify translation-related proteins in human spermatozoa and their association with cytoplasmic translation. The surface sensing of translation (SUnSET) method was used to measure translation activity in capacitated human and bovine spermatozoa. Two translation inhibitors, cycloheximide (CHX, cytoplasmic) and D-chloramphenicol (D-CP, mitochondrial) were used to identify which ribosomes were active in sperm. To spot newly synthesized proteins, puromycin-peptides were immunoprecipitated and analysed by mass spectrometry. A second approach was performed using translation inhibitors and analysing the sperm proteome by mass spectrometry. Bioinformatic analysis revealed that human spermatozoa possess 510 translation proteins, which were enriched for cytoplasmic mRNA translation. CHX decreased translation activity in mammalian sperm, whereas no effect was observed after D-CP treatment. Nine proteins were immunoprecipitated and identified as newly synthesized in capacitated bovine spermatozoa. CHX and D-CP decreased the level of 22 proteins that were replaced, or de novo translated during capacitation. New proteins were associated with relevant processes for sperm physiology. Both translation inhibitors decreased sperm rapid progressive motility and increased sperm immotility. Our results proved sperm translation occurs through cytoplasmic translation machinery in capacitated bovine and human spermatozoa. These results also support that sperm translation is required during capacitation to produce relevant proteins for sperm functions.

A randomized, placebo-controlled, quadruple-blind trial was performed to evaluate the effect of oral administration of the antioxidant combination Spermotrend® for three months on semen quality in infertile men with at least one abnormal variable in semen analysis. Eighty men were randomized between 2019 and 2022, receiving either the antioxidant combination Spermotrend® (n = 40, spermotrend-group) or placebo (n = 40, placebo-group). Although a total of 80 patients were enrolled in the study, the final data is only from 70 patients. The primary outcome measure was sperm motility (rapid progressive, progressive, and total motility). The values of primary and secondary outcomes between treatment initiation and treatment completion were compared within groups. Moreover, their changes between treatment initiation and treatment completion were compared between the placebo- and the spermotrend-groups. Sperm rapid progressive motility significantly increased in infertile men treated for three months with antioxidant combination Spermotrend® (+1.0%, 95% CI: 0.0 to +2.0, p = 0.04), while this increase was not observed in the placebo-group. Sperm progressive motility significantly increased in infertile men treated for three months with antioxidant combination Spermotrend® (+3.0%, 95% CI: 0.0 to +15.1, p = 0.02), while this increase was not observed in the placebo-group. Similarly, DFI was significantly decreased in infertile men treated for three months by antioxidant combination Spermotrend® (-3.2%, 95% CI: -5.8 to -0.5, p = 0.02). However, no statistically significant differences were observed in the changes of pre- and post-treatment values between the spermotrend- and the placebo-group regarding sperm progressive motility, concentration, normal morphology, DFI, and formation of 8-OH-dG. The antioxidant combination Spermotrend® appears to exert limited benefit on sperm motility and DFI in infertile men with at least one abnormal variable in semen analysis.

Currently, the potential role of obesity on semen quality remains unclear. In recent years, alternative anthropometric indices to body mass index (BMI), which can precisely distinguish body fat distribution, have been described to estimate central adiposity. However, the role of neck circumference (NC), a novel index of central adiposity, on semen quality is unknown. This study aims to reveal the potentials of adiposity indices, including NC, in predicting poor semen quality. A total of 4739 male participants between the ages of 17–55 were included in this cross-sectional study. Semen parameters, including sperm concentration, total count, total, progressive and rapid progressive motility and vitality, were divided into two categories according to the WHO classification for male infertility. As anthropometric measurements of obesity, BMI and central obesity indices including waist circumference (WC), hip circumference (HC), waist-to-hip ratio (WHpR), waist-to-height ratio (WHtR) and NC were examined. The predictive potentials of obesity indices for low semen parameters were evaluated by Receiver Operating Characteristics (ROC) curve analysis. The associations of obesity indices with semen parameters were analyzed by binary logistic regression analyze after adjusting potential confounding factors. Patients with lower semen parameters had higher obesity indices than those with normal parameters. All obesity indices have the predictive potentials for low semen parameters. After adjustment for confounders, the strongest associations were found between HC with sperm count parameters, WC and WHtR with sperm vitality and motility parameters and WHpR and NC with rapid progressive motility. Obesity plays an important role in male infertility. Body fat distribution appears to have specific roles on sperm functionality, which may influence different infertility markers. NC is a strong predictor for sperm rapid progressive motility, suggesting a role for male infertility.

This study evaluated the effects of chlorogenic acid (CGA) on human semen and on oxidative stress (OS) induced in vitro in human spermatozoa. After the treatment of the basal semen with 100 µM CGA, rapid and slow sperm progressive motility were evaluated and seminal F2-Isoprostanes (F2-IsoPs), a marker of OS, were quantified by ELISA. In a second set of experiments, semen was treated with 100 µM CGA, 1 mM H2O2 to induce OS, or H2O2+CGA; untreated samples were used as controls. Then, sperm motility, DNA integrity by the acridine orange test, F2-IsoPs and Nrf2 mRNA expression by RT-PCR were quantified. In CGA-treated specimens, rapid progressive sperm motility was increased (p < 0.01) and F2-IsoP levels decreased (p < 0.001) versus controls. The increase of F2-IsoP levels and DNA damage and the decrease of sperm motility after H2O2 treatment was reversed in the presence of CGA, which upregulated Nrf2 mRNA expression. These findings contributed to clarifying CGA's antioxidant activity and highlighted the positive impact of CGA on sperm progressive motility, suggesting also a possible mechanism of action based on the Nrf2 pathway. CGA can be useful during human semen handling procedures in the laboratory and in optimizing the recovery of motile spermatozoa through selection techniques during assisted reproductive technology protocols.

Cryopreservation of poultry sperm is crucial for preserving genetic diversity and protecting endangered breeds. Rooster sperm is highly sensitive to cryopreservation due to its high polyunsaturated fatty acid content, making it prone to damage during freezing and thawing. This study evaluated the effects of thawing temperatures and storage conditions on sperm quality, including motility, morphology, and viability. Frozen rooster semen samples were thawed at 37°C for 30 seconds, 60°C for 5 seconds, or 72°C for 5 seconds and stored at 4°C for up to 48 hours. Sperm quality parameters, including motility, kinematic characteristics, abnormal morphology, and viability, were assessed at 0, 3, 6, 9, 12, 24, and 48 hours using a Computer-Assisted Semen Analyzer (CASA). Post-thaw motility varied significantly between thawing temperatures at 24 and 48 hours (p < 0.05). Progressive and rapid progressive motility also differed significantly at 24 hours (p < 0.05). Sperm viability showed statistical differences across thawing groups at 24 and 48 hours (p < 0.05), while morphological abnormalities were significant at 12 and 48 hours (p < 0.05). Across all groups, sperm quality parameters varied significantly at each time point (p < 0.05). Thawing at 37°C and storing at 4°C for up to 24 hours optimizes sperm motility and viability, minimizing cryodamage and ensuring functional preservation. This approach is effective for short-term storage and crucial for sustaining genetic diversity and fertility in poultry breeding programs.

Infertility is a disease of the male or female reproductive system defined by the failure to achieve a pregnancy after 12 months or more of regular unprotected sexual intercourse. Infertility is estimated to affect 8%–12% of reproductive-age group couples (60–80 million couples) worldwide, with around 15–20 million (25%) in India alone. A significant contributor to male infertility is oligoasthenozoospermia, a term that encompasses both oligospermia and asthenozoospermia. Clinically, it is distinguished by reduced sperm concentration and motility. The prevalence of oligoasthenozoospermia among men has increased worldwide annually to 10%–15% under the influence of numerous harmful factors, such as environmental pollution, psychological stress, and unhealthy lifestyles, placing a significant burden on people and the social healthcare system. The drawbacks of the currently available treatments include their side effects, unclear clinical effectiveness, and increased cost. In Ayurveda, there is no direct correlation of oligoasthenozoospermia but it can be correlated with Ksheena shukra (~oligospermia) or Shukra kshaya (~reduced semen). It is a Vatapaittika-pradhana shukra dosha (~pathology due to the predominance of Vata-pitta dosha). This is a case report of a 36-year-old male complaining of infertility due to oligoasthenozoospermia. After 98 days of treatment, there is an increase in sperm count from 4 million/mL to 54 million/mL, rapid progressive motility from 7% to 35%, a decrease in nonprogressive motility from 22% to 5%, immotile sperm percentage from 71% to 55%. After one month of follow-up, the values were maintained and his partner achieved pregnancy. The observation of this case highlights the role of Vajeekarana aushadha (~aphrodisiac medicines) in the management of Oligoasthenozoospermia.

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.

The aim of this study was to evaluate using whey of cottage cheese as a diluting agent for ovine semen. Semen from five rams were collected, pooled, and divided into 3 parts, and diluted (1:20) using three solvents: Tris-15% egg yolk extender (E1, control), whey-cetric-10% egg yolk extender (E2), and whey alone (E3). The diluted semen was then allowed to equilibrate (5 oC for four hours), then frozen (-196 oC). After thawing, semen was assessed by Computer Assisted Semen Analyzers (CASA) program for progressive motility (PM), live sperm (LS), acrosome integrity (AI), abnormality (AB), and DNA damage of spermatozoa. Results showed that the percentages of LS, and AI in post-thawed semen were higher (P<0.50) and the proportion of chromatin damage was lower (P<0.05) with E2 than in other extenders (E1 and E3). Comparing E2 to other extenders, post-thawing semen examined by CASA showed improved (P<0.05) percentages of progressive, rapid progressive, and total motility, and decreased immotility. E2 showed significantly (P<0.05) higher VSL than in E1 and E3 with the highest LIN and STR percentages. E3 showed significantly (P<0.05) higher VCL and VAP than in E1 and E2 with the lowest LIN and STR percentages. Conclusion: Cottage whey-10% egg yolk can be used as an extender for ram semen cryopreservation. It increased the freezing ability of ram spermatozoa by improving most sperm characteristics as compared to Tris-egg yolk extender.