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Background: Sperm deoxyribonucleic acid (DNA) fragmentation is commonly encountered in spermatozoa, and the oocyte assumes responsibility for repairing sperm DNA fragmentation during the oocyte–embryo transition. Objectives: This study aimed to investigate whether the effect of sperm DNA fragmentation on intracytoplasmic sperm injection outcomes depends on the incidence of oocyte dimorphisms. Materials and methods: For the present cohort, 2942 fertilized oocytes from 525 patients submitted to intracytoplasmic sperm injection cycles were assessed. The present study was conducted in a private in vitro fertilization center affiliated to a university from June 2016 to July 2019. Semen samples were divided into the following two groups depending on the sperm DNA fragmentation index: a low fragmentation index group (<30% sperm DNA fragmentation, n = 1468) and a high fragmentation index group (≥30% sperm DNA fragmentation, n = 486). In addition, mature oocytes were examined before sperm injection, and intracytoplasmic and extracytoplasmic defects were recorded. The effect of the sperm DNA fragmentation index on laboratory and clinical intracytoplasmic sperm injection outcomes (depending on the presence of oocyte defects) was evaluated. Results: Significant increases in the rates of fertilization, high‐quality embryo, implantation, and pregnancy were noted for cycles with <30% sperm DNA fragmentation than cycles with ≥30% sperm DNA fragmentation (regardless of the presence of oocyte dimorphisms). The presence of dimorphisms significantly impacted laboratory and clinical outcomes. The lowest fertilization and high‐quality embryo rates were observed when a high sperm DNA fragmentation index was associated with the presence of dark cytoplasm, vacuoles, resistant membrane, and non‐resistant membrane. The lowest implantation and pregnancy rates were observed when a high sperm DNA fragmentation index was associated with the presence of vacuoles, defective perivitelline space, and fragmented polar body. The effect of sperm DNA fragmentation on miscarriage rates was significantly influenced by the presence of centrally located cytoplasmic granulation, a defective perivitelline space and non‐resistant membrane. Conclusion: A high sperm DNA fragmentation index increases the likelihood of miscarriage in intracytoplasmic sperm injection cycles, an effect that may potentially be magnified by the presence of oocyte dysmorphisms. [ABSTRACT FROM AUTHOR]

Background: Sperm DNA fragmentation was hypothesized to have a role in the pathogenesis of recurrent pregnancy loss. Unfortunately, the quality of already published evidence is low. Objectives: To investigate the association between sperm DNA fragmentation and idiopathic recurrent pregnancy loss by limiting, as much as possible, the interference of confounding factors. Materials and methods: This was a retrospective multicenter case–control study conducted in two Italian University Hospitals (i.e., Policlinico Gemelli, Rome and Humanitas S. Pio X, Milan) from July 2020 to March 2022. Cases were men belonging to couples affected by first trimester idiopathic recurrent pregnancy loss, defined as the previous loss of two or more pregnancies. Two control groups were selected: (i) men belonging to couples with proven fertility (i.e., at least two previous full‐term pregnancies) (control group A); (ii) men belonging to couples with proven infertility (i.e., the failure to achieve a pregnancy after 12 months or more of regular unprotected sexual intercourse) (control group B). The sperm DNA fragmentation index was measured by the terminal deoxynucleotidyl transferase dUTP nick end labeling assay. Results: We included 74 cases, 37 men with proven fertility (control group A) and 100 men belonging to infertile couples (control group B). The median sperm DNA fragmentation index was significantly lower in control group A (17%, interquartile range: 14.3%–20.6%) compared to both case group (24.5%, interquartile range: 17%–32%; p < 0.0001) and control group B (24%, interquartile range: 18.9%–30%; p = 0.001). The rate of subjects with sperm DNA fragmentation index greater than 30% was significantly higher in both case groups (28%, 95% confidence interval [18%–40%]) and control group B (26%, 95% confidence interval [18%, 36%]) compared to control group A (0%, 95% confidence interval [0%–10%]) (p < 0.001). Multivariate regression models yielded a significant association between sperm DNA fragmentation index and recurrent pregnancy loss (adjusted odds ratio 1.13, 95% confidence interval [1.04–1.23], p = 0.006), but failed to show an association between sperm DNA fragmentation index and infertility (adjusted odds ratio 1.13, 95% CI [1–1.29], p = 0.05). Conclusions: Men within couples affected by recurrent pregnancy loss or infertility had a significantly higher rate of sperm DNA fragmentation compared to fertile controls. However, after adjusting for covariates, sperm DNA fragmentation index was associated only with recurrent pregnancy loss. [ABSTRACT FROM AUTHOR]

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

Background: Cigarette smoking seems to have a negative impact on men's reproductive health, but our knowledge of its effects on the reproductive function of Russian men is still very limited. The purpose of this study was to evaluate the effect of cigarette smoking on semen quality, including sperm DNA fragmentation, hormonal, zinc and metabolic status in young men from the general multi-ethnic Russian population (n=1,222, median age 23 years) and to find out the ethno-specific effects of smoking by comparing male groups of different ethnicity. Methods: Each participant filled out a standardized questionnaire, provided one blood and semen sample. Semen parameters, serum reproductive hormones, lipids, glucose, uric acid and seminal zinc were analyzed. Participants were classified as smokers (n=450) and non-smokers (n=772), and smokers were stratified into moderate (<10 cigarettes/day) and heavy (>10 cigarettes/day) smokers. Results: In the entire study population, heavy smokers were characterized by a decrease in semen volume, total sperm count, sperm concentration and motility, and an increase in sperm DNA fragmentation and teratozoospermia compared with non-smokers (p<0.05). There was also a reduction in the serum and seminal zinc level as well as an impairment in metabolic health in smokers compared with non-smokers (p<0.05). No significant differences between smokers and non-smokers were found for serum levels of LH, FSH, inhibin B, testosterone and estradiol. In the second part of our study, the most numerous ethnic groups of Slavs (n=654), Buryats (n=191), and Yakuts (n=125) were selected from the entire study population. Among three ethnic groups, the smoking intensity was higher in Slavs than in Buryats or Yakuts suggesting a greater tobacco addiction in Slavs than in Asians. A decrease in semen parameters and seminal zinc levels, and an increase in sperm DNA fragmentation and teratozoospermia was observed only in smoking Slavs (p<0.05); moderate decrease in testosterone and increase in triglyceride levels were revealed in smoking Yakuts (p<0.05), but no significant changes were detected in smoking Buryats. Conclusion: We concluded that cigarette smoking has an ethno-specific effect on male reproductive function, probably due to the different activity of the seminal antioxidant system, which is yet to be elucidated. [ABSTRACT FROM AUTHOR]

Background Methods Results Conclusion Patient summary Recurrent pregnancy loss (RPL), defined as two or more failed clinical pregnancies, affects 1%–3% of couples trying to conceive. Nowadays up to 50% of cases remain idiopathic. In this context, paternal factors evaluation is still very limited. The aim is to address the topic of the male factor in RPL with a broad approach, analyzing collectively data on sperm DNA fragmentation (SDF) and semen parameters. We systematically searched in Pubmed/MEDLINE and Google Scholar from inception to February 2023. A protocol has been registered on PROSPERO (ID number CRD42022278616). PRISMA guidelines were followed.Pooled results from 20 studies revealed a higher DNA fragmentation rate in the RPL group compared to controls (mean difference [MD] 9.21, 95% CI 5.58–12.85, p < 0.00001, I2 98%). Age, body mass index (BMI), smoking, and alcohol intake were not associated with DNA fragmentation. Subgroup analysis by different SDF assays (TUNEL and COMET at a neutral pH vs. indirect assessment with other assays) and ethnicity did not highlight different results (p = 0.25 and 0.44).Results pooled from 25 studies showed a significant difference comparing RPL and control groups regarding ejaculation volume (MD −0.24, 95% CI −0.43; −0.06, p 0.01, I2 66%), total sperm number (MD −10.03, 95% CI −14.65; −5.41, p < 0.0001, I2 76%), total sperm motility (MD −11.20, 95% CI −16.15; −6.25, p < 0.0001, I2 96%), progressive sperm motility (MD −7.34, 95% CI −10.87; −3.80, p < 0.0001, I2 97%), and normal sperm morphology (MD −5.99, 95% CI −9.08; −2.90, p 0.0001, I2 98%). A sub‐analysis revealed that Asian and Africans, but not white‐European RPL men had lower progressive sperm motility compared to controls.In conclusion, current review and meta‐analysis findings suggested that SDF and some specific semen parameters were associated with RPL in a multi‐ethnic evaluation. This effort opens future direction on a growing awareness of, first, how the male factor plays a key role and, second, how appropriate would be to establish a direct dialogue between the gynecologist and the urologist.We performed a systematic review and meta‐analysis on the male component of RPL. We found that sperm DNA fragmentation and some specific sperm parameters are significantly associated with RPL. [ABSTRACT FROM AUTHOR]

To date, infertility affects 10% to 15% of couples worldwide. A male factor is estimated to account for up to 50% of cases. Oral supplementation with antioxidants could be helpful to improve sperm quality by reducing oxidative damage. At the same time, there is a growing interest in the literature on the use of testicular sperm in patients with high DNA fragmentation index (DFI). This narrative review aims to evaluate the effectiveness of supplementation of oral antioxidants in infertile men with high DFI compared to testicular sperm retrieval. The current evidence is non-conclusive because of serious risk of bias due to small sample sizes and statistical methods. Further large well-designed randomised placebo-controlled trials are still required to clarify the exact role of these to different therapeutic approaches. [ABSTRACT FROM AUTHOR]

Benzo(a)pyrene (BaP) is considered one of the most dangerous air pollutants for adverse health effects, including reproductive toxicity. It is found both in male and female reproductive fluids likely affecting spermatozoa after the selection process through cervical mucus, a process mimicked in vitro with the swim-up procedure. In vitro effects of BaP (1, 5, 10 µM) were evaluated both in unselected and swim-up selected spermatozoa after 3 and 24 h of incubation. BaP reduced total, progressive and hyperactivated motility and migration in a viscous medium both in swim-up selected and unselected spermatozoa. Viability was not significantly affected in swim-up selected but was reduced in unselected spermatozoa. In swim-up selected spermatozoa, increases in the percentage of spontaneous acrosome reaction and DNA fragmentation were observed after 24 h of incubation, whereas no differences between the control and BaP-treated samples were observed in caspase-3 and -7 activity, indicating no effects on apoptotic pathways. ROS species, evaluated by staining with CellROX® Orange and Dihydroethidium, did not differ in viable spermatozoa after BaP treatment. Conversely, the percentage of unviable ROS-positive spermatozoa increased. Our study suggests that BaP present in male and female genital fluids may heavily affect reproductive functions of human spermatozoa. [ABSTRACT FROM AUTHOR]

Background: There is ongoing research to find an optimum modality to predict male fertility potential. Aims: To compare the semen parameters, sperm DNA damage and seminal metal levels of Zinc, Lead and Aluminium among the male partners of couples with unexplained infertility and men with proven fertility. Settings and Design: Prospective case-control study at a tertiary level teaching hospital. Materials and Methods: One hundred male partners of couples with unexplained subfertility and 50 men with proven fertility were included in the study. Male partners of unexplained infertility couples and fertile men were compared for their semen parameters, sperm DNA Fragmentation Index (DFI) and seminal metal levels in semen. Statistical Analysis Used: Chi-square test, Student's t-test, sensitivity and specificity analysis, binomial logistic regression analysis. Results: Fertile men had statistically significantly higher mean progressive sperm motility than male partners of unexplained infertility (53.12 ± 9.89% vs. 44.81 ± 19.47%, P = 0.005). Semen volume and sperm concentration were comparable among the cases and control population. The mean sperm DFI was significantly lower among fertile men (10.83 ± 6.28 vs. 21.38 ± 10.28, P < 0.0001). Plotting the receiver-operating characteristic curve the threshold for discrimination was calculated to be 18% DFI. The sensitivity specificity and overall accuracy were 43%, 84% and 56.67%, respectively when the DFI cut-off was set at 18%. Zinc concentration in the semen had a strong positive correlation (Point Biserial correlation coefficient = 0.831) with fertility, whereas lead and aluminium had a moderate negative correlation. Conclusion: Conventional semen analysis had limited differentiating ability for unexplained infertility. The sperm DFI may be employed for explanatory purposes among couples with unexplained subfertility. A lower discriminatory threshold of DFI (18%) has better overall accuracy as opposed to a 30% cutpoint for unexplained subfertility. Among metals, Zinc was strongly correlated with fertility status. [ABSTRACT FROM AUTHOR]

Background/Aim: Sperm quality has experienced a decline in recent years, with this issue being particularly pronounced in industrialized nations, suggesting a potential link to occupational exposures. Evaluating sperm DNA fragmentation can yield valuable insights into male fertility, although its association with occupational exposures remains less well-established. Our study aimed to investigate the relationship between self-reported occupational exposures and sperm DNA fragmentation in infertile men. Methods: This retrospective cohort study involved 391 infertile men who sought fertility treatment at a university clinic between 2017 and 2020. A brief questionnaire was administered to collect data on patients’ demographic characteristics, medical history, occupation, and exposure types. In this comparative study, patients were categorized into two groups based on their occupational exposures (the unexposed and exposed groups). The exposed group was further sub-grouped according to their specific exposure types, which included cement, solvents, metals, pesticides, mechanical vibration, and heat. The primary outcome in this study was assessed using the terminal deoxynucleotidyl transferase-mediated nick end-labeling test (TUNEL), with results expressed as the sperm DNA fragmentation index (DFI). Results: Patients in the exposed group exhibited a significantly higher sperm DFI compared to those in the unexposed group (14 [17] vs. 8 [9], P<0.001). After accounting for potential confounding factors, our results demonstrated that several occupational exposure factors significantly increased the risk of elevated sperm DFI (>15%) levels, including solvents (odds ratio (OR) 8.2, 95% confidence interval (CI) 3.6–18.5, P<0.001), metals (OR: 2.2, 95% CI: 1.0–4.7, P=0.048), pesticides (OR: 14.6, 95% CI: 1.6–130.7, P=0.016), mechanical vibration (OR: 2.6, 95% CI: 1.5–4.6, P<0.001), and heat (OR: 6.4, 95% CI: 1.7–23.5, P=0.005). Conclusion: The findings of our study corroborate earlier research suggesting that occupational exposures may have adverse effects on sperm DNA fragmentation in men. The identification and management of such exposures as part of routine clinical practice could offer a complementary approach to enhancing infertility treatment outcomes. [ABSTRACT FROM AUTHOR]

Antioxidant therapy should be reserved for infertile patients who actually exhibit signs of oxidative stress (OS). Nevertheless, there is no consensus regarding the measure of the primary endpoint and the assay that should be used. The formation of 8-hydroxy-2′-deoxyguanosine (8-OHdG), an early marker of sperm DNA oxidation (SDO), was analyzed using flow cytometry, in men at a University hospital setup for infertility treatment. Similar to conventional semen parameters, 8-OHdG assay was validated on fresh semen samples to reduce the variability of results. SDO was associated with semen volume, sperm concentration, leucocytes and round cells, but not with age, body mass index, sperm DNA fragmentation (SDF) or OS. Whether the semen samples were normal or subnormal according to the WHO criteria, the expression of 8-OHdG was not different. Receiver operating characteristic curve analysis could discriminate two independent populations. Both SDF and SDO were independently expressed. A high SDF did not reveal a high SDO and vice versa. The thresholds for SDO have been established, but vary with the techniques used. The methodology for SDO needs to be further validated and optimized on a larger clinically defined patient population before the outcome measure is fit to monitor antioxidant therapy in male infertility. [ABSTRACT FROM AUTHOR]

Introduction. Lifestyle factors, including smoking, alcohol (AU) and drug use (DU), can affect male fertility. The aim of the study was to investigate the frequency and characteristics of smoking, AU and DU in men from infertile couples, to identify the most significant associations of disorders of spermatogenesis and social risk factors.Material and methods. At the 1st stage of the study, the case histories of 1198 men from infertile couples were analyzed to determine the frequency of smoking, AU and DU, at the 2nd stage, a more detailed questionnaire of 239 patients from the general sample was conducted for detailed characteristics of smoking and/or AU and/or DU, at the 3rd stage, a comprehensive analysis of ejaculate from men who consumed strong alcohol, but did not smoke or use DU (n = 46) was performed in comparison with men without bad habits taken into account (n = 60).Results. In the general sample, the frequency of AU was 73 %, smoking – 41 %, DU –17 % (mostly in anamnesis). 47.9 % of AU men consumed beer (1.5 (1–2.5) liters per week, 25.6 % – strong alcohol (250 (100–500) ml per week), 7.6 % – champagne/wine (500 (250–725) ml per week); 92.7 % of smokers used cigarettes, 7.3 % – electronic cigarettes (smoking experience was 15 (10–20) years, the number of cigarettes per day was 15 (10–20) pieces); more than 90 % of DU men have a history of non-injection DU. Men who consumed only strong alcohol, but did not smoke and did not use drugs, increased sperm DNA fragmentation compared to men without bad habits taken into account: 16.0 (13.5–19.6) and 12.8 (8.8–19.4) %, respectively, p = 0.018.Conclusions. Among men from infertile couples, a high frequency of AU was revealed with the predominant use of beer or strong alcohol, the use of the latter increases sperm DNA fragmentation; smoking is characterized by a long experience; active DU is rare. Given the specifics of the requirements for the state of reproductive health of men applying to reproductive medicine centers, it is important to have knowledge about the frequency and severity of smoking, AU and DU among them, as well as about the relationship of risk factors of infertility with the parameters of ejaculate.

Objective: To determine the effects of sperm deoxyribonucleic acid (DNA) fragmentation at the time of fertilization on in vitro fertilization (IVF) outcomes and genetic diagnosis using next generation sequencing. Design: Prospective double-blinded study. Setting: Private Clinic. Patients: Couples (n = 150). Intervention: In vitro fertilization with preimplantation genetic testing for aneuploidy and sperm DNA fragmentation assay, as in sperm chromatin structure assay the day of retrieval. Main Outcome Measures: Laboratory outcomes are listed in the results section. Statistical analysis was performed using JMP, XYLSTAT, and STATA version 15. Results: The sperm DNA fragmentation index (DFI) in the neat ejaculate did not predict fertilization rate, quality, blastulation, or genetic diagnosis. No statistically significant results were obtained comparing 15%, 20%, 30% except for DFI. No statistically significant differences in oocyte source age or male age were observed. No statistically significant differences comparing 15%, 20%, 30% DFI at the time of standard IVF or intracytoplasmic sperm injection (ICSI) were observed for % euploid, aneuploid, mosaic, blastulation, biopsied, or D5/total biopsied. The DFI of >15% had more good quality D3 embryos than the 20% group compared with the

Abstract Background The evaluation of the infertile couple is often complex as multiple factors in both the male and female can contribute, including social history. Previous studies have displayed that male ethanol consumption can disturb sperm motility, nuclear maturity, and deoxyribonucleic acid (DNA) integrity. The main purpose of this study is to evaluate the effects of male alcohol use on sperm chromatin structure analysis (SCSA®). This study was a retrospective chart review of 209 couples that presented to a midsize infertility clinic in the Midwest and had a semen analysis and SCSA® performed. Data extracted from the electronic medical record included demographics, tobacco use, alcohol use, occupational exposures, semen analysis results, and SCSA® results (DNA Fragmentation index (DFI) and High DNA stainability (HDS)). Statistical analysis was performed on this data set to determine significance with a p-level of 0.05, with the primary input being level of alcohol use and primary outcome being the SCSA® parameters. Results Overall, 11% of the cohort had heavy alcohol use (> 10 drinks/week), 27% moderate (3–10/week), 34% rare (0.5- 10% (a marker of immature sperm chromatin). Level of alcohol use was not significantly associated with HDS > 10% or DFI. Heavier alcohol use was significantly associated with lower sperm count (p = 0.042). Increasing age was significantly associated with increasing DNA Fragmentation Index (p = 0.006), increased sperm count (p = 0.002), and lower semen volume (p = 0.022). Exposure to heat at work was significantly associated with lower semen volume (p = 0.042). Tobacco use was associated with lower sperm motility (p

Abstract Background We analyzed the sperm DNA fragmentation index (DFI) and general semen test based on the World Health Organization (WHO) criteria and compared the two tests using semen factors. In addition, we examined whether DFI is a reliable parameter associated with in vitro fertilization (IVF) outcomes. Methods Sperm chromatin dispersion (SCD) and general semen tests were conducted in accordance with the WHO 2010 guidelines, and correlations between the two tests were investigated. The WHO criteria were set as the cutoff values for each of the following factors: semen volume, concentration, total sperm count, motility, and normal morphology, and compared with the DFI results. Results The subjects had a mean sperm DFI of 15.3% ± 12.6%, and the DFI increased with age. In contrast, motility and normal morphology decreased as the DFI increased. Patients who satisfied the WHO criteria in terms of concentration, total sperm count, and motility had a significantly lower DFI than those who did not satisfy the criteria. Therefore, evaluation with a general semen test based on the WHO criteria should be regarded as a qualitative evaluation of all factors other than semen volume and normal morphology. Conclusions High DFI (≥ 30%) caused a low blastocyst development rate following intracytoplasmic sperm injection. Male infertility due to DFI should be suspected when IVF results are poor despite normal semen findings based on the WHO criteria. The results of this study suggest that the SCD test may more accurately evaluate the correlation between IVF clinical outcomes and male infertility. Therefore, it is important to focus on DFI measurements.

BackgroundCigarette smoking seems to have a negative impact on men’s reproductive health, but our knowledge of its effects on the reproductive function of Russian men is still very limited. The purpose of this study was to evaluate the effect of cigarette smoking on semen quality, including sperm DNA fragmentation, hormonal, zinc and metabolic status in young men from the general multi-ethnic Russian population (n=1,222, median age 23 years) and to find out the ethno-specific effects of smoking by comparing male groups of different ethnicity.MethodsEach participant filled out a standardized questionnaire, provided one blood and semen sample. Semen parameters, serum reproductive hormones, lipids, glucose, uric acid and seminal zinc were analyzed. Participants were classified as smokers (n=450) and non-smokers (n=772), and smokers were stratified into moderate (≤10 cigarettes/day) and heavy (>10 cigarettes/day) smokers.ResultsIn the entire study population, heavy smokers were characterized by a decrease in semen volume, total sperm count, sperm concentration and motility, and an increase in sperm DNA fragmentation and teratozoospermia compared with non-smokers (p

To date, infertility affects 10% to 15% of couples worldwide. A male factor is estimated to account for up to 50% of cases. Oral supplementation with antioxidants could be helpful to improve sperm quality by reducing oxidative damage. At the same time, there is a growing interest in the literature on the use of testicular sperm in patients with high DNA fragmentation index (DFI). This narrative review aims to evaluate the effectiveness of supplementation of oral antioxidants in infertile men with high DFI compared to testicular sperm retrieval. The current evidence is non-conclusive because of serious risk of bias due to small sample sizes and statistical methods. Further large well-designed randomised placebo-controlled trials are still required to clarify the exact role of these to different therapeutic approaches.

Sperm cells age during storage in the male reproductive organs, which can lead ejaculate quality to decline dramatically over time. Such may pose a particular concern in species with constrained sperm supplies, where adult males cannot replenish their sperm reserves. In eusocial hymenopterans (i.e. ants, bees and wasps with permanent castes), males typically reach adulthood with a limited quantity of sperm, and their testes deteriorate shortly thereafter. In adult life, both sexes mate during a single reproductive event. While males die quickly after copulation, they persist posthumously via sperm stored in their mates' spermatheca, sometimes for decades. As a result of this mating system, males should experience intense selection pressure to produce and transfer high‐quality sperm. Using the black garden ant (Lasius niger) as a model system, we investigated whether the duration of storage in the accessory testes before copulation affected sperm quality. We found that there was no impact of male age on sperm number or viability and that sperm DNA fragmentation decreased rather that increased with age. These results highlight the extent to which male ants, and possibly other taxa with constrained sperm supplies, have evolved extremely specialized strategies to ensure reproductive performance. [ABSTRACT FROM AUTHOR]

Several preventive measures, including vaccination, have been implemented owing to the severe global effect of coronavirus disease 2019 (COVID-19), but there is still limited evidence in the effect of this disease and vaccination against it on male fertility. Therefore, this study is to compare sperm parameters of infertile patients with or without COVID-19 infection and the effect of COVID-19 vaccine types on them. Semen samples of infertile patients were collected consecutively at Universitas Indonesia - Cipto Mangunkusumo Hospital (Jakarta, Indonesia). COVID-19 was diagnosed by rapid antigen or polymerase chain reaction (PCR) tests. Vaccination was performed with three types of vaccine, namely inactivated viral vaccine, messenger RNA (mRNA) vaccine, and viral vector vaccine. Spermatozoa were then analyzed on the World Health Organization recommendations, and DNA fragmentation was assayed with the sperm chromatin dispersion kit. The results showed that the COVID-19 group experienced a significant decrease in sperm concentration and progressive motility (both P < 0.05), but there was no significant change in morphology or sperm DNA fragmentation index (DFI; both P > 0.05). The viral vector vaccine caused a decrease in morphology as well as an increase in DFI compared with the control (both P < 0.05), meanwhile results for those who were vaccinated with the inactivated and mRNA types were not significant compared with the control (both P > 0.05). We conclude that COVID-19 has negative effects on sperm parametes and sperm DNA fragmentation, and we found that the viral vector vaccines affect sperm parameter values and DNA fragmentation negatively. Further studies with a larger population and longer follow-up are needed to confirm the results. [ABSTRACT FROM AUTHOR]

Purpose: 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. Methods: Semen samples from 239 men were collected. CA0 under different incubation intervals (5, 10, 30, and 60 min) 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). Results: Total motility and motile sperm concentration increased in a time- and temperature-dependent manner and the total motility peaked for 30 min 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). Conclusion: 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. [ABSTRACT FROM AUTHOR]

BACKGROUND It has long been thought that the factors affecting embryo and foetal development were exclusively maternally derived; hence, if issues regarding fertility and embryo development were to arise, the blame has traditionally been placed solely on the mother. An escalating interest in how paternal factors influence embryo development, however, has begun to prove otherwise. Evidence suggests that both seminal plasma (SP) and sperm contribute multiple factors that shape embryogenesis. This review thus focuses on the role that semen has in driving early embryonic development, and describes how paternal factors, such as SP, sperm centriole, sperm proteins, sperm RNA, sperm DNA, and its integrity, together with epigenetics, may influence the female reproductive tract and post-fertilization events. The important contributions of paternal factors to embryo development highlight the imperative need for further research in this area, which is sure to bring forth breakthroughs leading to improvements in infertility diagnosis and ART as well as reducing the risk of miscarriage. OBJECTIVE AND RATIONALE This review provides a comprehensive overview of the role of human semen in development of the early embryo, with the aim of providing a better understanding of the influence of SP and sperm on early embryonic divisions, gene and protein expression, miscarriage, and congenital diseases. SEARCH METHODS PubMed searches were performed using the terms 'sperm structure', 'capacitation', 'acrosome reaction', 'fertilization', 'oocyte activation', 'PLCζ', 'PAWP', 'sperm-borne oocyte activation factor', 'oocyte activation deficiency', 'sperm centriole', 'sperm transport', 'sperm mitochondria', 'seminal plasma', 'sperm epigenetics', 'sperm histone modifications', 'sperm DNA methylation', 'sperm-derived transcripts', 'sperm-derived proteins', 'sperm DNA fragmentation', 'sperm mRNA', 'sperm miRNAs', 'sperm piRNAs', and 'sperm-derived aneuploidy'. The reviewed articles were restricted to those published in English between 1980 and 2022. OUTCOMES The data suggest that male-derived factors contribute much more than just the male haploid genome to the early embryo. Evidence indicates that semen contributes multiple factors that help shape the fate of embryogenesis. These male-derived factors include contributions from SP, the paternal centriole, RNA and proteins, and DNA integrity. In addition, epigenetic changes have an impact on the female reproductive tract, fertilization, and early stages of embryo development. For example, recent proteomic and transcriptomic studies have identified several sperm-borne markers that play important roles in oocyte fertilization and embryogenesis. WIDER IMPLICATIONS This review highlights that several male-derived factors are required to work in tandem with female counterparts to allow for correct fertilization and development of the early embryo. A deeper understanding of the contributions of paternal factors that are shuttled over from the sperm cell to the embryo can shed light on how to improve ART from an andrological perspective. Further studies may aid in preventing the passing on of genetic and epigenetic abnormalities of paternal origin, thus decreasing the incidence of male factor infertility. In addition, understanding the exact mechanisms of paternal contribution may assist reproductive scientists and IVF clinicians in determining new causes of recurrent early miscarriage or fertilization failure. [ABSTRACT FROM AUTHOR]

Background and Aims: Varicocele is one of the most common causes of male infertility in which testicular function is progressively damaged. This study aims to investigate the effect of varicocelectomy and mast cell stabilizers on reducing sperm DNA fragmentation in infertile patients with varicocele. Methods: In this randomized clinical trial, after obtaining ethical approval and informed consent, infertile patients with varicocele were randomly assigned to three groups: varicocelectomy, 1 mg ketotifen, and varicocelectomy plus with 1 mg ketotifen for 3‐month follow‐up. Semen analysis and estimation of DNA fragmentation index (DFI) were obtained from all people in the studied time periods. Results: In this study, 420 infertile patients with varicocele were included with an average age of 31.6 years. The results showed that sperm morphology, sperm motility, sperm count, and sperm DFI had significant differences after the intervention (p < 0.05). Also, the results of semen analysis for each group showed that there was a significant improvement in sperm morphology, motility, count, and DFI index of infertile people with varicocele after the intervention (p < 0.05). But the difference was statistically higher in the varicocelectomy plus with 1 mg ketotifen group than in the other groups (p < 0.01). Conclusion: The evidence and results of this study showed that the use of varicocelectomy plus ketotifen in infertile people with varicocele was more effective in improving Semen parameters than the use of mast cell stabilizers or varicocelectomy alone. [ABSTRACT FROM AUTHOR]

Several preventive measures, including vaccination, have been implemented owing to the severe global effect of coronavirus disease 2019 (COVID-19), but there is still limited evidence in the effect of this disease and vaccination against it on male fertility. Therefore, this study is to compare sperm parameters of infertile patients with or without COVID-19 infection and the effect of COVID-19 vaccine types on them. Semen samples of infertile patients were collected consecutively at Universitas Indonesia - Cipto Mangunkusumo Hospital (Jakarta, Indonesia). COVID-19 was diagnosed by rapid antigen or polymerase chain reaction (PCR) tests. Vaccination was performed with three types of vaccine, namely inactivated viral vaccine, messenger RNA (mRNA) vaccine, and viral vector vaccine. Spermatozoa were then analyzed on the World Health Organization recommendations, and DNA fragmentation was assayed with the sperm chromatin dispersion kit. The results showed that the COVID-19 group experienced a significant decrease in sperm concentration and progressive motility (both P < 0.05), but there was no significant change in morphology or sperm DNA fragmentation index (DFI; both P > 0.05). The viral vector vaccine caused a decrease in morphology as well as an increase in DFI compared with the control (both P < 0.05), meanwhile results for those who were vaccinated with the inactivated and mRNA types were not significant compared with the control (both P > 0.05). We conclude that COVID-19 has negative effects on sperm parametes and sperm DNA fragmentation, and we found that the viral vector vaccines affect sperm parameter values and DNA fragmentation negatively. Further studies with a larger population and longer follow-up are needed to confirm the results.

Purpose: Seminal oxidative stress (OS) is a recognized factor potentially associated with male infertility, but the efficacy of antioxidant (AOX) therapy is controversial and there is no consensus on its utility. Primary outcomes of this study were to investigate the effect of AOX on spontaneous clinical pregnancy, live birth and miscarriage rates in male infertile patients. Secondary outcomes were conventional semen parameters, sperm DNA fragmentation (SDF) and seminal OS. Materials and Methods: Literature search was performed using Scopus, PubMed, Ovid, Embase, and Cochrane databases. Only randomized controlled trials (RCTs) were included and the meta-analysis was conducted according to PRISMA guidelines. Results: We assessed for eligibility 1,307 abstracts, and 45 RCTs were finally included, for a total of 4,332 infertile patients. We found a significantly higher pregnancy rate in patients treated with AOX compared to placebo-treated or untreated controls, without significant inter-study heterogeneity. No effects on live-birth or miscarriage rates were observed in four studies. A significantly higher sperm concentration, sperm progressive motility, sperm total motility, and normal sperm morphology was found in patients compared to controls. We found no effect on SDF in analysis of three eligible studies. Seminal levels of total antioxidant capacity were significantly higher, while seminal malondialdehyde acid was significantly lower in patients than controls. These results did not change after exclusion of studies performed following varicocele repair. Conclusions: The present analysis upgrades the level of evidence favoring a recommendation for using AOX in male infertility to improve the spontaneous pregnancy rate and the conventional sperm parameters. The failure to demonstrate an increase in live-birth rate, despite an increase in pregnancy rates, is due to the very few RCTs specifically assessing the impact of AOX on live-birth rate. Therefore, further RCTs assessing the impact of AOX on live-birth rate and miscarriage rate, and SDF will be helpful.

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This retrospective study assessed the biological intra-individual variability of the percentage of sperm with DNA damage (SDF) observed in subsequent ejaculates of the same individual. Variation in SDF was analyzed using the Mean Signed Difference (MSD) statistic based on 131 individuals, comprising 333 ejaculates. Either two, three or four ejaculates were collected from each individual. With this cohort of individuals two main questions were addressed; (1) does the number of ejaculates analyzed influence the variability in the level of SDF associated with each individual? and (2) is the variability observed in SDF similar when individuals are ranked according to their level of SDF? Results showed that the variation observed in mean SDF was not different when 2, 3 or 4 ejaculates were analyzed; consequently, we suggest that the assessment of SDF based on two ejaculates is likely to be representative of the mean SDF expected for the individual. In parallel, it was determined that the variation in SDF increased as SDF increased; in individuals presenting with an SDF value of lower than 30% (potentially fertile), only 5% possessed levels of MSD that could be considered as variable as that presented by individuals presenting with a recurrent high SDF. Finally, we showed that a single assessment of SDF in individuals with medium SDF (20-30%) was less likely to be predictive of the SDF value in the next ejaculate, and therefore, less informative of the patient's SDF status. [ABSTRACT FROM AUTHOR]

Cancer impairs spermatogenesis, whereas results on sperm DNA integrity are controversial and no data are available about sperm oxidative stress. In cancer patients, we detected sperm DNA fragmentation (sDF) and both viable (ROS production in viable sperm fraction/viable spermatozoa) and total (ROS production in viable sperm fraction/total spermatozoa) oxidative stress. We found that cancer (22.50 (17.00–26.75)%, n = 85) increased sDF with respect to the control groups in both normozoospermic subfertile patients (NSP) (12.75 (8.63–14.88)%, n = 52, p < 0.001) and in healthy donors (HD) (8.50 (7.00–14.00)%, n = 19, p < 0.001). The induction of viable oxidative stress (n = 96) with cancer was even higher: 36.60 (24.05–58.65)% versus 11.10 (8.63–14.90)% in NSP (p < 0.001) and 9.60 (8.00–14.03)% in HD (p < 0.001). Similar, albeit lower, differences were found for total oxidative stress. SDF sharply correlated to viable oxidative stress when we considered all subjects (cancer patients and controls) (r = 0.591, p < 0.001, n = 134), but no correlation was found when only cancer patients were studied (r = 0.200; p > 0.05, n = 63). In conclusion, cancer significantly increases sDF and sperm oxidative stress levels. Additional mechanisms to oxidative attack might be responsible for increased sDF in cancer patients. Because sperm oxidative stress might affect the outcomes of sperm cryopreservation, of cancer treatments and of sperm epigenoma, the detection of oxidative stress could be of help in managing the reproductive issues of cancer patients. [ABSTRACT FROM AUTHOR]

Chromosomal polymorphisms are structural variations in chromosomes that define the genomic variance of a species. These alterations are recurrent in the general population, and some of them appear to be more recurrent in the infertile population. Human chromosome 9 is highly heteromorphic, and how its rearrangement affects male fertility remains to be fully investigated. In this study, we aimed to investigate the association between the polymorphic rearrangements of chromosome 9 and male infertility via an Italian cohort of male infertile patients. Cytogenetic analysis was carried out, along with Y microdeletion screening, semen analysis, fluorescence in situ hybridization, and TUNEL assays using spermatic cells. Chromosome 9 rearrangements were observed in six patients: three of them showed a pericentric inversion, while the others showed a polymorphic heterochromatin variant 9qh. Of these, four patients exhibited oligozoospermia associated with teratozoospermia, along with a percentage of aneuploidy in the sperm of above 9%, in particular, an increase in XY disomy. Additionally, high values for sperm DNA fragmentation (≥30%) were observed in two patients. None of them had microdeletions to the AZF loci on chromosome Y. Our results suggest that polymorphic rearrangements of chromosome 9 might be associated with abnormalities in sperm quality due to incorrect spermatogenesis regulation. [ABSTRACT FROM AUTHOR]

Background: We analyzed the sperm DNA fragmentation index (DFI) and general semen test based on the World Health Organization (WHO) criteria and compared the two tests using semen factors. In addition, we examined whether DFI is a reliable parameter associated with in vitro fertilization (IVF) outcomes. Methods: Sperm chromatin dispersion (SCD) and general semen tests were conducted in accordance with the WHO 2010 guidelines, and correlations between the two tests were investigated. The WHO criteria were set as the cutoff values for each of the following factors: semen volume, concentration, total sperm count, motility, and normal morphology, and compared with the DFI results. Results: The subjects had a mean sperm DFI of 15.3% ± 12.6%, and the DFI increased with age. In contrast, motility and normal morphology decreased as the DFI increased. Patients who satisfied the WHO criteria in terms of concentration, total sperm count, and motility had a significantly lower DFI than those who did not satisfy the criteria. Therefore, evaluation with a general semen test based on the WHO criteria should be regarded as a qualitative evaluation of all factors other than semen volume and normal morphology. Conclusions: High DFI (≥ 30%) caused a low blastocyst development rate following intracytoplasmic sperm injection. Male infertility due to DFI should be suspected when IVF results are poor despite normal semen findings based on the WHO criteria. The results of this study suggest that the SCD test may more accurately evaluate the correlation between IVF clinical outcomes and male infertility. Therefore, it is important to focus on DFI measurements. [ABSTRACT FROM AUTHOR]

Abstract Background and Aims Varicocele is one of the most common causes of male infertility in which testicular function is progressively damaged. This study aims to investigate the effect of varicocelectomy and mast cell stabilizers on reducing sperm DNA fragmentation in infertile patients with varicocele. Methods In this randomized clinical trial, after obtaining ethical approval and informed consent, infertile patients with varicocele were randomly assigned to three groups: varicocelectomy, 1 mg ketotifen, and varicocelectomy plus with 1 mg ketotifen for 3‐month follow‐up. Semen analysis and estimation of DNA fragmentation index (DFI) were obtained from all people in the studied time periods. Results In this study, 420 infertile patients with varicocele were included with an average age of 31.6 years. The results showed that sperm morphology, sperm motility, sperm count, and sperm DFI had significant differences after the intervention (p

In this clinical era of intracytoplasmic sperm injection (ICSI), where a single spermatozoon is chosen for fertilization, the diagnostic functionality of the classical parameters typically associated with fertilization, such as sperm concentration, sperm motility, acrosome integrity, and mitochondria, is perhaps becoming less critical. In contrast, the contribution of sperm DNA quality to our understanding of the impact of male fertility within the context of ICSI is gaining increasing interest and importance. Even with respect to natural conception, high levels of sperm DNA fragmentation (SDF) in the ejaculate can adversely affect reproductive outcomes. However, the precise origin of SDF pathology in sperm cells is often ambiguous and most likely to be multifactorial. Hence, the genetic makeup of an individual, unbalanced REDOX processes, enzymatic activity, environmental and lifestyle factors, and even damage during sperm handling in the laboratory all operate in a unique and often synergistic manner to produce or induce sperm DNA damage. Surprisingly, the contribution of active enzymes as potential agents of SDF has received much less attention and, therefore, is likely to be underrated. This review highlights the roles of different enzymes related to the degradation of sperm DNA as possible effectors of DNA molecules in spermatozoa.

Introducción: El plomo no tiene función fisiológica, por lo que cualquier nivel encontrado en el organismo es potencialmente peligroso. Por las importantes consecuencias adversas que ejerce sobre el hombre que pueden ser provocadas por su exposición ocupacional, se hace énfasis en sus efectos sobre la salud reproductiva masculina. Objetivo: Informar sobre aspectos generales relacionados con los efectos tóxicos del plomo en el hombre, su diagnóstico, fuentes de contaminación ocupacional y su relación con la fertilidad masculina. Método: Búsqueda en la literatura publicada en diversas bases de datos y portales bibliográficos. Del total de artículos encontrados, 64 fueron elegidos por su relevancia para ser incluidos en esta revisión, siendo el 65 % de estas publicaciones de la última década. Desarrollo: Múltiples estudios se refieren a los efectos adversos del plomo, incluso a niveles menores de 5 μg/dL. Su toxicidad reproductiva se evidencia en la disminución de la calidad del semen, daño a la estructura de los espermatozoides y en la integridad de la membrana espermática. Estudios publicados muestran que la medición de la cantidad de plomo en el tracto reproductivo puede ser un buen indicador para evaluar sus efectos sobre la fertilidad masculina. En Cuba la minería, la fundición secundaria, la fabricación, reacondicionamiento y reciclado de baterías Pb-ácido son sectores de alta exposición ocupacional, por lo que se debe reforzar la vigilancia de salud, promover la salud reproductiva y fomentar cambios de estilo de vida, para contribuir a la reducción de la exposición de estos trabajadores. Conclusiones: Existen evidencias de que la exposición ocupacional crónica al plomo, afecta negativamente la salud, la calidad espermática y el índice de fragmentación espermático. No existen en Cuba estudios que evalúen la calidad seminal y la fragmentación del ADN espermático en trabajadores expuestos ocupacionalmente a plomo Introduction: Lead has no physiological function; therefore, any level of it found in the body is potentially dangerous. Due to the significant adverse consequences that it can cause to any man subjected to occupational exposure to this metal, its effects on male reproductive health are emphasized. Objective: To inform about general aspects related to the toxic effects of lead in men, its diagnosis, sources of occupational contamination, and its relationship with male fertility. Method: A search was carried out in the literature published in various databases and bibliographic portals. Of the total number of found articles, 64 were selected for their relevance to be included in this review. 65% of the texts were published in the recent decade. Development: Multiple studies refer to the adverse effects of lead, even at levels under 5 μg/dL. Its reproductive toxicity is evidenced by decreased semen quality, as well as by damage to the spermatozoa structure and the integrity of the spermatic membrane. Published studies show that measurement of the amount of lead in the reproductive tract can be a good indicator to assess its effects on male fertility. In Cuba, mining, secondary smelting, manufacturing, reconditioning, and recycling of Pb-acid batteries are sectors with high occupational exposure to lead, a reason why health surveillance must be strengthened, reproductive health must be promoted, and changes in lifestyle must be encouraged, in order to contribute to reducing the exposure of these workers. Conclusions: There is evidence that chronic occupational exposure to lead affects health, sperm quality and sperm fragmentation index negatively. There are no studies in Cuba that evaluate seminal quality and sperm DNA fragmentation in workers occupationally exposed to lead

Sperm competition is a pervasive evolutionary force that shapes sperm traits to maximize fertilization success. Indeed, it has been shown to increase sperm production in both vertebrates and invertebrates. However, sperm production is energetically costly, which may result in trade-offs among sperm traits. In eusocial hymenopterans, such as ants, mating dynamics impose unique selective pressures on ejaculate. Males are sperm limited: they enter adulthood with a fixed amount of sperm that will not be renewed. We explored whether sperm competition intensity was associated with sperm quantity and quality (i.e. sperm viability and DNA fragmentation) in nine Cataglyphis desert ants. Our results provide phylogenetically robust evidence that sperm competition is positively correlated with sperm production and sperm viability. However, it was unrelated to sperm DNA integrity, indicating the absence of a trade-off involving this trait. These findings underscore that sperm competition may strongly mould sperm traits and drive reproductive performance in eusocial Hymenoptera. [ABSTRACT FROM AUTHOR]

In March 2020, the Italian government imposed a national lockdown which was almost completely removed in June 2020. Due to the abrupt stop of human activities, emissions of air pollutants decreased. Air pollution is an environmental risk factor for noncommunicable disease and mortality. Emerging evidence also suggests a role in male infertility. In this study, we compared sperm DNA fragmentation (sDF) levels and conventional semen parameters between subjects undergoing sDF determination and routine semen analysis in a single Italian centre, during about 6 months before (N = 119) and after lockdown (N = 105). After lockdown, we found an improvement of sperm progressive motility (48.00[38.50–58.00]% vs. 42.00[33.00–53.00]%) and sDF levels (as total: 24.79[18.33–33.97]% vs. 35.02[25.04–45.73]%, p <.001; brighter: 14.02[10.69–17.93]% vs 18.54[13.58–25.82]%, p <.001 and dimmer sDF: 9.24[5.64–15.78]% vs. 12.24[8.08–19.10]%, p <.01), mirrored by a decrease of leukocyte semen concentration (p <.01). The improvement of sperm motility and DNA quality was maintained after adjusting for leukocyte concentration and several conditions known to affect sperm motility and/or sDF levels. With a significant decrease in air pollution observed in Tuscany during and after lockdown, associated improvement in sperm motility and DNA quality in patients referred to the infertility clinic is suggestive of the potential role of air pollution in male infertility. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the relationship between DNase activity associated with bacterial contamination of incubated bovine frozen-thawed spermatozoa and elevated sperm DNA fragmentation. Electrophoresis analysis of plasmid PBR322 incubated for 30 min at 37 °C with the supernatant of the diluent of frozen-thawed centrifuged bovine semen straws infected with bacteria showed clear evidence of DNase activity when compared to plasmid incubated in similarly prepared non-infected bovine diluent supernatant (Experiment 1). This DNase activity was subsequently found to be time dependent (0–60 min) and its activity prevented in the presence of EDTA (10 and 20 mM; Experiment 2). Semen straws infected (n = 10) and not infected (n = 10) with bacteria where incubated at 37 °C for up to 48h post-thaw. Semen infected with bacteria showed an exponential increase in bacterial growth and a corresponding increase in sperm DNA fragmentation. Non-infected semen samples showed no change in the incidence of sperm DNA fragmentation over the same period of incubation (Experiment 3). Our experiments reinforce the idea that exogenous DNases present in the semen should be considered as one of the primary contributing causes of sperm DNA fragmentation post ejaculation. In the case of the bull, post-thaw incubation of commercial straws contaminated with bacteria, resulted in increased levels of sperm DNA fragmentation, most likely associated with DNase activity (potentially restriction endonucleases) derived from the bacteria. Such adverse changes in sperm DNA fragmentation, as described here in vitro, may be also operative after insemination in the female reproductive tract (in vivo) and highlight the importance of implementing high levels of hygiene practice during semen processing, especially in light of future trends of bacterial resistance to the common antibiotics used in semen diluents. • This study determined whether DNase activity associated with bacterial infection in post-thaw bovine semen caused sperm DNA fragmentation (SDF). • We showed bacterial DNase activity of the supernatant of post-thawed bovine semen on plasmid DNA was time dependent but could be prevented by EDTA. • Bacterial DNase activity associated with the supernatant of post-thawed bovine semen resulted in elevated levels of SDF. • Adverse changes in SDF associated with bacterial DNase activity highlight the importance of hygiene during semen processing. [ABSTRACT FROM AUTHOR]

What is the effect of a novel non-centrifugation method (Io-Lix) of sperm selection on sperm parameters and intracytoplasmic sperm injection (ICSI) reproductive outcomes? This pilot study elevated the capacity of the Io-Lix sperm selection protocol to improve sperm parameters (concentration, motility and sperm DNA fragmentation) of the neat ejaculate. Once established, the reproductive outcomes of Io-Lix selected spermatozoa were used for autologous and donor oocyte ICSI programmes and their efficacy compared with those using conventional swim-up. Io-Lix sperm selection resulted in lower sperm concentration yield (P < 0.001) and sperm DNA fragmentation (P < 0.001) but higher sperm motility (P < 0.001) when compared with spermatozoa in the neat ejaculate. When compared with swim-up sperm selection the Io-Lix protocol resulted in a 14.7% (P = 0.028) increase in pregnancy rate and 16.3% (P = 0.047) reduction in miscarriages in the autologous ICSI programme. A similar comparison of sperm selection procedures employed for a donor oocyte ICSI programme showed no difference in terms of their respective reproductive outcomes. The Io-Lix sperm selection protocol resulted in improved pregnancy rate and reduction in miscarriage when applied to autologous ICSI, which was attributed to a reduction in the proportion of spermatozoa with DNA damage post-selection. A similar finding was not apparent in the donor oocyte programme, which may be associated with the capacity of the donor oocyte to repair sperm DNA post-syngamy. [ABSTRACT FROM AUTHOR]

Semen parameters are unable to inform on the function or fertilizing capacity of the male gamete. Standardized methods are provided by the WHO but, the lower reference limits have reduced sensitivity to predict chances of conception. Subfertile men may be falsely classified as "normal" and a male factor contributing to genome instability may be overlooked. Semen parameters, sperm DNA fragmentation (SDF), sperm chromatin maturity and stability, and sperm aneuploidy were assessed in fertile (F), subfertile normozoospermic (SN) and subfertile non-normozoospermic males (SN-N). Standardized assays employing flow cytometry were used to detect genome instability. Sperm DNA fragmentation did not differ significantly whether the semen samples were from a fertile (F), subfertile normozoospermic (SN) or subfertile non-normozoospermic male (SN-N). Chromatin decondensation was significantly reduced and hyperstability significantly increased in the SN group as compared to the F group. The frequency of diploidy was significantly different in the three study groups with significance between F and SN and between F and SN-N groups. Subfertile men with normal semen parameters are often excluded from extensive genetic testing. Genome instability might be an independent attribute of semen quality detecting problems not seen with semen analysis alone. [ABSTRACT FROM AUTHOR]

Sperm separation plays a critical role in assisted reproductive technology. Based on migration, density gradient centrifugation and filtration, a properly selected sperm could help in increasing assisted reproductive outcomes in teratozoospermia (TZs). The current study aimed to assess the prognostic value of four sperm selection techniques: density gradient centrifugation (DGC), swim-up (SU), DGC-SU and DGC followed by magnetic-activated cell sorting (DGC-MACS). These were evaluated using spermatozoa functional parameters. A total of 385 infertile couples underwent the procedure of intracytoplasmic sperm injection (ICSI), with an isolated teratozoospermia in the male partner. Semen samples were prepared by using one of the mentioned sperm preparation techniques. The improvements in the percentage of normal mature spermatozoa, rate of fertilization, cleavage, pregnancy and the number of live births were assessed. The normal morphology, spermatozoa DNA fragmentation (SDF) and chromatin maturity checked by using chromomycin A3 (CMA3) with DGC-MACS preparation were better compared to the other three methods. Embryo cleavage, clinical pregnancy and implantation were better improved in the DGC-MACS than in the other tested techniques. The DGC-MACS technique helped in the selection of an increased percentage of normal viable and mature sperm with intact chromatin integrity in patients with teratozoospermia. [ABSTRACT FROM AUTHOR]

Introduction: Infertility affects one in every six couples in developed countries, and approximately 50% is of male origin. In 2021, sperm DNA fragmentation (SDF) testing became an evidence-based test for fertility evaluations depicting fertility more clearly than standard semen parameters. Therefore, we aimed to summarize the potential prognostic factors of a higher SDF. Methods: We conducted a systematic search in three medical databases and included studies investigating any risk factors for SDF values. We calculated mean differences (MD) in SDF with 95% confidence interval (CI) for exposed and non-exposed individuals. Results: We included 190 studies in our analysis. In the group of associated health conditions, varicocele (MD = 13.62%, CI: 9.39–17.84) and impaired glucose tolerance (MD = 13.75%, CI: 6.99–20.51) had the most significant increase in SDF. Among malignancies, testicular tumors had the highest impact, with a maximum of MD = 11.3% (CI: 7.84–14.76). Among infections, the overall effects of both Chlamydia and HPV were negligible. Of lifestyle factors, smoking had the most disruptive effect on SDF – an increase of 9.19% (CI: 4.33–14.06). Different periods of sexual abstinence did not show significant variations in SDF values. Age seemed to have a more drastic effect on SDF from age 50 onwards, with a mean difference of 12.58% (CI: 7.31–17.86). Pollution also had a detrimental effect – 9.68% (CI: 6.85–12.52). Conclusion: Of the above risk factors, varicocele, impaired glucose tolerance, testicular tumors, smoking, pollution, and paternal age of over 50 were associated with the highest SDF. Trial registration: CRD42021282533. [ABSTRACT FROM AUTHOR]

The intracytoplasmic sperm injection (ICSI) technique was invented to solve severe male infertility due to altered sperm parameters. Nowadays, it is applied worldwide for the treatment of couple infertility. ICSI is performed with any available spermatozoon from surgery or ejaculated samples, whatever are the sperm motility, morphology or quantity. The aim of the present review was to study if embryo development and kinetics would be modified by (1) ICSI under the technical aspects, (2) the micro-injected spermatozoa in connection with male infertility. From published data, it can be seen that ICSI anticipates the zygote kinetics Furthermore, because fertilization rate is higher in ICSI compared to conventional in vitro fertilization (IVF), more blastocysts are obtained for clinical use in ICSI. Sperm and spermatozoa characteristics, such as sperm parameters, morphology and vitality, DNA content (levels of sperm DNA fragmentation, microdeletions, and chromosomal abnormalities), RNA content, epigenetics, and sperm recovery site (testicular, epididymis, and ejaculated), have an impact on fertilization and blastocyst rates and embryo kinetics in different ways. Even though ICSI is the most common solution to solve couples' infertility, the causes of male infertility are crucial in building a competent spermatozoa that will contribute to normal embryonic development and healthy offspring. [ABSTRACT FROM AUTHOR]

Sperm deoxyribonucleic acid (DNA) damage has recently emerged as one of the most controversial topics in male reproductive medicine. While level I evidence indicates that abnormal sperm DNA damage has substantial adverse effects on reproductive outcomes (including chance of pregnancy and risk of miscarriage), there is limited consensus on how sperm DNA fragmentation (SDF) testing should be performed and/or interpreted in clinical practice. In this article, we review: (1) how SDF is assessed, (2) cumulative evidence regarding its impact on reproductive outcomes, (3) methods for mitigating high SDF, and (4) the most recent practice guidelines available for clinicians regarding the use and interpretation of SDF testing. [ABSTRACT FROM AUTHOR]

The evaluation of morpho-functional sperm characteristics alone is not enough to explain infertility or to predict the outcome of Assisted Reproductive Technologies (ART): more sensitive diagnostic tools are needed in clinical practice. The aim of the present study was to analyze Sperm DNA Fragmentation (SDF) and sperm-borne miR-34c-5p and miR-449b-5p levels in men of couples undergoing ART, in order to investigate any correlations with fertilization rate, embryo quality and development. Male partners (n = 106) were recruited. Semen analysis, SDF evaluation and molecular profiling analysis of miR-34c-5p and miR-449b-5p (in 38 subjects) were performed. Sperm DNA Fragmentation evaluation- a positive correlation between SDF post sperm selection and the percentage of low-quality embryos and a negative correlation with viable embryo were found. SDF > 2.9% increased the risk of obtaining a non-viable embryo by almost 4-fold. Sperm miRNAs profile—we found an association with both miRNAs and sperm concentration, while miR-449b-5p is positively associated with SDF. Moreover, the two miRNAs are positively correlated. Higher levels of miR-34c-5p compared to miR-449b-5p increases by 14-fold the probability of obtaining viable embryos. This study shows that SDF, sperm miR-34c-5p, and miR-449b-5p have a promising role as biomarkers of semen quality and ART outcome. [ABSTRACT FROM AUTHOR]

Background: Protamines are the nuclear proteins essential for chromatin compaction during spermatogenesis. Spermatozoa may develop the SDF if the spermatogenesis is dysregulated, which affects male infertility Objective: This study was conducted to quantify the PROTAMINE 1 gene expression in the semen specimen of infertile Iraqi male and estimate sperm DNA fragmentation. Patients and Material: 50 semen specimens were gathered from infertile males and 50 samples from healthy fertile individual as control group and subjected for semen analysis, determinations the expression of PROTAMINE 1 gene and estimation sperm DNA fragmentation. Results: There were no appreciable differences in the ejaculation volume between the studied groups. The fertile group's mean sperm concentration (m/ml)(49.58 ±2.93)was statistically significantly (p <0.05) higher than the infertile male group(27.08±15.7). The progressive motility of fertile male(57.20 ± 2.27) was significantly(P < 0.001) higher as compared to other infertile male groups(22.29 ± 1.95). The morphologically normal sperm of control males(52.20 ± 1.91) was a highly significant (P < 0.001) as compared to infertile male groups (36.30± 2.20). The fold protamine 1 gene expression of patients group(0.26) had a lower significantly than the healthy group (1). The mean SDF of infertile male group(43.05 ± 2.93) was statistically significant (P < 0.001)than the fertile male group(17.97 ± 9.90). Conclusion: The current study found that increased sperm with breaks DNA was linked to reduced protamine 1 gene expression and negative impacts on sperm parameters. [ABSTRACT FROM AUTHOR]

Background: Sperm DNA fragmentation has been proposed as a candidate test for the assessment of sperm function on the premise that damage to the sperm chromatin is associated with a detrimental reproductive outcome. The objective of our study was to investigate whether sperm DNA fragmentation testing has a prognostic value, and thus can play a pivotal role in selecting future patients for intra-uterine insemination (IUI) therapy. Methods: This was a prospective cohort study conducted in a University Hospital setting. SDF was measured through TUNEL assay on the fresh semen sample presented at diagnosis and at insemination in couples with idiopathic/mild male infertility undergoing natural cycle IUI treatment. The generalized estimating equation (GEE)-model and multivariable model were used to analyze the probability of live birth and clinical pregnancy, respectively. ROC analysis was carried out to determine an SDF cut-off. Results: There was an inverse relationship between SDF in the ejaculate of the diagnostic semen sample and CP (p = 0.02; OR 0.94 95% CI (0.90, 0.989)) as well as LB (p = 0.04; OR 0.95 95% CI (0.90, 0.9985)). No significant association was found between SDF after gradient and IUI outcome in the diagnostic sample nor between SDF (ejaculate/after gradient) in the IUI samples. The ROC analysis proposed a cutoff of 17.5% as the best compromise between sensitivity and specificity in the diagnostic SDF for live birth; however, the test diagnostics are low, with an AUC of 0.576. Conclusions: Overall, this study strengthens the hypothesis of an inverse relationship between SDF and CP/LB. Furthermore, SDF taken together with other clinical characteristics might provide more insight into male reproductive potential and predicting IUI outcome. Couples with SDF ≥ 17.5% in the diagnostic semen sample did not reach live birth. Further research is necessary to establish the diagnostic and prognostic potential of SDF as an add-on test. [ABSTRACT FROM AUTHOR]

The origin and quality of gametes are likely to influence the kinetics of embryonic development. The purpose of the study was to assess the impact of sperm nuclear quality, and in particular sperm chromatin condensation, on the kinetics of early embryo development after intracytoplasmic sperm injection (ICSI). Our study included 157 couples who benefitted from ICSI for male factor infertility. Chromatin condensation and DNA fragmentation were assessed in spermatozoa prior to ICSI. Above the 20% threshold of sperm condensation defect, patients were included in the abnormal sperm chromatin condensation (ASCC) group; below the 20% threshold, patients were included in the normal sperm chromatin condensation (NSCC) group. After ICSI, the oocytes were placed in the time-lapse incubator. The kinetics of the cohort's embryonic development have been modeled. The fading times of pronuclei and the time to two blastomeres (t2, first cleavage) and four blastomeres (t4, third cleavage) differed significantly between the NSCC and ASCC groups, with earlier events occurring in the ASCC group. On the other hand, the state of sperm chromatin condensation did not seem to have an impact on live birth rates or the occurrence of miscarriages. The kinetics of early embryonic development was accelerated in males with a sperm chromatin condensation defect without compromising the chances of pregnancy or promoting miscarriage. However, our study highlights the paternal contribution to early embryonic events and potentially to the future health of the conceptus. [ABSTRACT FROM AUTHOR]

Background: Approximately 8%–12% of couples worldwide face infertility, with infertility of individuals assigned male at birth (AMAB) contributing to at least 50% of cases. Conventional semen analysis commonly used to detect sperm abnormalities is insufficient, as 30% of AMAB patients experiencing infertility show normal results in this test. From a genetic perspective, the assessment of sperm DNA fragmentation (SDF) is important as a parameter of sperm quality. Methods: In this narrative study, we review and discuss pathophysiological causes, DNA repair mechanisms, and management of high SDF. We then summarize literature exploring the association between SDF and reproductive outcomes. Main Findings: Recent systematic reviews and meta‐analyses have revealed a significant association between high SDF in AMAB individuals and adverse reproductive outcomes including embryo development, natural conception, intrauterine insemination, and in vitro fertilization. However, the association with live birth rates and pregnancy rates following intracytoplasmic injection remains inconclusive. The disparities among quantitative assays, inconsistent reference range values, absent high‐quality prospective clinical trials, and clinical heterogeneity in AMAB patients with elevated SDF represent the main limitations affecting SDF testing. Conclusion: The evaluation and management of SDF plays an important role in a subset of AMAB infertility, but widespread integration into clinical guidelines will require future high‐quality clinical trials and assay standardization. [ABSTRACT FROM AUTHOR]

Background: It is well known that cryopreservation procedure may result in negative impact on spermatozoa function throughout relative overproduction of reactive oxygen species (ROS). However, ROS overproduction can be corrected by antioxidants. Objective: To find out the effect of vitamin A on cryopreserved spermatozoa throughout measuring their motility rate and DNA fragmentation and compare it with vitamin E effect. Methods: Forty seminal fluid specimens were individually collected from forty healthy, non-drug and nonalcohol consumers, normozoospermic males in Um Al-Baneen Center for Infertility Management and In Vitro Fertilization in Baghdad. To facilitate comparing between used vitamins and experimenting each one alone, those specimens were distributed into 4 groups of 10 specimens each. Following deriving their own controls without any vitamin treatment, specimens of these groups, in general, were treated with two concentrations of vitamin A (20 μg/dl and 30 μg/dl), and two concentrations of vitamin E (10 μmol/l and 20 μmol/l). Then, each specimen was incubated for 1 hour before being cryopreserved in liquid nitrogen for 14 days. Motility percentage and DNA fragmentation were assessed following cryopreservation and thawing of spermatozoa. Results: Results revealed that there were significant statistically differences in post-thawing motility and DNA fragmentation means between specimens treated with vitamin A and their relevant control; between specimens treated with vitamin E and their relevant control; and between specimens treated with vitamin E and those treated with vitamin A among all groups of the study. Conclusion: These results lead to the conclusion that both of vitamin A and E play an important role in improving and protecting sperm motility and DNA integrity following cryopreservation, and vitamin E is more effective than vitamin A. [ABSTRACT FROM AUTHOR]

Purpose: Seminal oxidative stress (OS) is a recognized factor potentially associated with male infertility, but the efficacy of antioxidant (AOX) therapy is controversial and there is no consensus on its utility. Primary outcomes of this study were to investigate the effect of AOX on spontaneous clinical pregnancy, live birth and miscarriage rates in male infertile patients. Secondary outcomes were conventional semen parameters, sperm DNA fragmentation (SDF) and seminal OS. Materials and Methods: Literature search was performed using Scopus, PubMed, Ovid, Embase, and Cochrane databases. Only randomized controlled trials (RCTs) were included and the meta-analysis was conducted according to PRISMA guidelines. Results: We assessed for eligibility 1,307 abstracts, and 45 RCTs were finally included, for a total of 4,332 infertile patients. We found a significantly higher pregnancy rate in patients treated with AOX compared to placebo-treated or untreated controls, without significant inter-study heterogeneity. No effects on live-birth or miscarriage rates were observed in four studies. A significantly higher sperm concentration, sperm progressive motility, sperm total motility, and normal sperm morphology was found in patients compared to controls. We found no effect on SDF in analysis of three eligible studies. Seminal levels of total antioxidant capacity were significantly higher, while seminal malondialdehyde acid was significantly lower in patients than controls. These results did not change after exclusion of studies performed following varicocele repair. Conclusions: The present analysis upgrades the level of evidence favoring a recommendation for using AOX in male infertility to improve the spontaneous pregnancy rate and the conventional sperm parameters. The failure to demonstrate an increase in live-birth rate, despite an increase in pregnancy rates, is due to the very few RCTs specifically assessing the impact of AOX on live-birth rate. Therefore, further RCTs assessing the impact of AOX on live-birth rate and miscarriage rate, and SDF will be helpful. [ABSTRACT FROM AUTHOR]

Sperm DNA fragmentation (SDF) that occurs during the freezing–thawing of sperm may negatively impact the treatment outcomes of assisted reproductive technologies (ART). In a previous study, we developed a human sperm cryopreservation reagent containing carboxylated poly-L-lysine (CPLL) that reduced SDF after freeze-thawing compared with clinically popular cryopreservation reagents containing human serum albumin. However, it is unclear whether CPLL reduces SDF, as it differed from the constituents of the commercial cryopreservation reagents used for comparison. Therefore, here, we examined whether CPLL reduces the SDF of human sperm and evaluated reactive oxygen species (ROS) levels and lipid peroxidation (LPO), which are the causes of SDF; mitochondrial injury, ROS production; and impaired sperm motility. Furthermore, optimal antioxidants and their concentrations that could further enhance the reduction in SDF were determined for future clinical application in ART and underwent the same functional evaluations. CPLL can reduce SDF via inhibition of intracytoplasmic ROS and LPO. Furthermore, the addition of 0.1 mM resveratrol avoided the enhancement of SDF, which potentially affects mitochondrial and cytoplasmic ROS and LPO. This novel human sperm cryopreservation reagent containing CPLL and resveratrol has the potential to improve treatment outcomes in ART using frozen sperm.