Epidemiological, Clinical, and Biomarker Profile of Male Infertility in Morocco: A Retrospective Single-Center Study of 1399 Cases

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

Many cycling women with elevated basal FSH level have been discouraged from undergoing IVF treatment. This is because elevated basal FSH is associated with poorer assisted reproduction treatment outcome. It has been argued that high FSH reflects not only reduced ovarian reserve but also poor oocyte quality. The aim of this study is to assess the value of treating cycling women who have elevated basal FSH and to assess the reasons for the reduction in both pregnancy rate (PR) and live birth rate (LBR). Between January 1997 and December 2001, 2057 patients underwent 3401 consecutive IVF/ICSI cycles in which the basal level of FSH (days 2-4) was determined at an earlier cycle. Analysis, however, was only performed for a single cycle per patient. All cases were divided into four cohorts according to FSH levels: group A, FSH <10 IU/ml; group B, 10.1-15 IU/ml; group C, 15.1-20 IU/ml; and group D, FSH >20 IU/ml. Each group was stratified further into subgroups according to age, < or =38 and >38 years. Both PR (A, 32.3%; B, 19.8%; C, 17.5%; and D, 3%) and LBR (A, 24.7%; B, 13.2%; C, 13.8%; and D, 3%) were significantly reduced in the higher FSH level groups. LBR was significantly higher in the younger subgroups (A, 32.2%; B, 21.8%; C, 20%; and D, 16.7%) as compared with the older subgroups (A, 12.1%; B, 8.3%; C, 10.5%; and D, 0%). Higher levels of FSH were significantly associated with more cycle cancellation, a larger amount of gonadotrophin required to achieve follicular maturity, and a lower number of eggs collected, embryos available and embryos transferred. In all cases, however, there was no significant correlation between FSH levels and fertilization rate or miscarriage rate. Younger cycling women with elevated FSH had significantly higher LBR compared with older women with normal FSH (21.2% versus 12.1%). Furthermore, the cumulative LBR after three cycles in these younger patients with elevated FSH levels was 49.3%. Although there is a reduction in both PR and LBR associated with higher levels of basal FSH, it is clear that in cycling women, high basal FSH is not a contraindication to IVF treatment, and a respectable PR and LBR can be achieved especially in young women. The reduction in PR and LBR is due to reduced reserve rather than poor oocyte quality. Clinics refusing to treat cycling women with elevated basal FSH levels may be denying these women a reasonable, albeit low, chance of achieving a birth with their own genetic material. Clinicians should use basal FSH levels as a guide to advise patients about their chances of achieving a live birth, not to exclude patients with a predicted lower success rate from a treatment programme.

Processing of semen can result in increased sperm DNA fragmentation

Introduction: Varicocele, characterized as a palpable, dilated, and tortuous pampiniform plexus of testicular veins, is present in approximately 35% of men with primary infertility. While the exact mechanisms underlying varicocele-associated infertility remain unclear, evidence indicates reduced sperm concentration, impaired motility, oxidative stress, and increased sperm DNA fragmentation (SDF) as contributing factors. This study prospectively evaluates the effects of varicoceles on conventional semen parameters and SDF. Materials and Methods: Sixty semen samples from men with (30) and without (30) varicocele were analyzed to assess semen parameters and SDF. Varicocele presence was clinically evaluated and confirmed through color Doppler sonography. Semen parameters were assessed according to the World Health Organization 2021 6th edition guidelines, with sperm concentration measured using a Makler counting chamber and computer-assisted semen analysis. SDF was evaluated in both groups using the sperm chromatin dispersion test (Halosperm kit). Results for semen parameters and DNA fragmentation index were compared between the varicocele and nonvaricocele groups. Results: The study evaluates the impact of varicocele on fertility, lifestyle habits, semen parameters, and DNA fragmentation. Primary infertility was more prevalent in both groups, affecting 63.3% of varicocele and 56.7% of nonvaricocele patients. Lifestyle analysis showed significantly higher smoking (33.3%, P = 0.010) and multiple habits (26.6%, P = 0.039) in the varicocele group, while nonvaricocele patients were more likely to have no habits (56.6%, P = 0.001). Semen analysis revealed significant impairments in varicocele patients, including lower sperm count, total sperm number, total motility, nonprogressive motility, and normal morphology (P &lt; 0.05). DNA fragmentation was significantly higher in varicocele patients, with 73.3% showing &gt;30% fragmentation compared to 40% in nonvaricocele patients (P = 0.009). Normal semen parameters were observed in only 13.3% of varicocele patients compared to 60% in nonvaricocele patients (P = 0.001). Conclusions: This study highlights that infertile men with varicoceles exhibit significant abnormalities in semen quality, coupled with elevated levels of sperm nuclear DNA fragmentation, suggesting the critical role of varicoceles in male infertility.

Recently a polymorphic variant of the FSH receptor in which amino acid asparagine (Asn) at position 680 is replaced by serine (Ser) was found. This is associated with higher FSH levels in the early follicular phase and an increased FSH requirement to obtain follicular response in IVF patients. The aim of our study was to test the hypothesis that this receptor isoform occurs more often in regularly menstruating subfertility patients with elevated basal FSH than in those with normal early follicular phase FSH. A retrospective cohort study of 38 subfertility patients with a regular menstrual cycle and elevated FSH (>10 IU/l) compared to 40 patients with normal early follicular phase FSH was carried out. DNA was analysed to determine the FSH receptor genotype. The N680S variant on one or both alleles of the FSH receptor gene was significantly more prevalent in patients with elevated FSH (P < 0.05). The homozygous Asn/Asn variant at codon 680 was found in 45% of women with normal FSH and in 21% of women with elevated FSH. The homozygous Ser/Ser receptor variant was present in 12.5% of women with normal FSH and in 21% of patients with elevated FSH. Also the heterozygous combination of both variants Asn/Ser occurred more often in women with elevated FSH (58 versus 42.5%). The N680S sequence variation of the FSH receptor is found in >75% of the cases with elevated basal FSH and suggests a higher FSH threshold.

To investigate the relationship among seminal oxidation-reduction potential (nORP), sperm DNA fragmentation (DFI) and semen parameters in patients with varicocele. Clinical data of 522 patients treated in the reproductive andrology clinic of the Northern Theater General Hospital from November 2023 to December 2023 were retrospectively analyzed, including 435 men of childbearing age and 87 men of infertile age. The patients were divided into the varicocele group (n=116) and non-varicocele group (n=406) according to clinical diagnosis. The differences of seminal plasma nORP, DFI, sperm high DNA stain ability (HDS) and semen parameters were analyzed between the two groups. The relationship among general clinical data, seminal plasma nORP, semen parameters, DFI and HDS in patients with varicocele were further analyzed. According to the severity of varicocele, the patients were divided into three groups, including mild, moderate and severe. And the differences of seminal plasma nORP and semen parameters, DFI and HDS among all groups were analyzed. The differences of seminal plasma nORP, semen parameters, DFI and HDS were compared between the varicocele and non-varicocele groups. The total sperm count, sperm concentration, progressive motility sperm percentage (PR%) and normal sperm morphology rate (NSMR) in patients with varicocele were significantly lower than those in control group (P<0.05). And seminal plasma nORP, DFI and HDS in patients with varicocele were significantly higher than those in control group (P<0.05). Seminal plasma nORP in patients with varicocele was significantly negatively correlated with total sperm, sperm concentration and NSMR (P<0.05), and significantly positively correlated with DFI and HDS (P<0.05). There were significant differences in nORP, total sperm count, sperm concentration, PR%, DFI and HDS among mild, moderate and severe varicocele groups (P<0.05). Seminal plasma nORP, sperm concentration, PR% and DFI in severe group were significantly lower than those in mild and moderate groups(P<0.05). Sperm count and HDS in severe group were significantly lower than those in mild group (P<0.05). In infertile patients, seminal plasma nORP, DFI and HDS in varicocele group were significantly higher than those in control group (P<0.05). And PR% in varicocele group was significantly lower than that in control group (P<0.05). Seminal plasma nORP in patients with varicocele may be an important marker of oxidative stress affecting DFI and semen parameters.

Relationship of environmental exposure temperature and temperature extremes on sperm DNA fragmentation index in men with different BMI values and the indirect effect of DNA fragmentation index on semen parameters

Alcohol consumption has commonly been associated with semen parameters. However, the association between alcohol intake and semen parameters in primary and secondary infertile men remains unclear. In this study, 776 infertile men from China were grouped according to alcohol intake: abstainers, moderate drinkers (<9 units/week, up to approximately 100 g of ethanol) and heavy drinkers (≥9 units/week). Semen parameters, including semen volume, sperm concentration, total sperm count, progressive motility and normal morphology were investigated. Alcohol consumption and other lifestyle factors were assessed by questionnaire. Logistic regression models were applied. There was no significant association between alcohol consumption and semen parameters in men with primary infertility. Smaller testis volumes and lower sperm concentrations were found among moderate and heavy drinkers in the secondary infertility group than among abstainers. After adjustment for potential confounders, men with secondary infertility and heavy alcohol consumption had a higher risk of abnormal sperm concentrations (OR = 3.72; 95% CI, 1.04, 13.37). These findings suggest that alcohol intake may decrease sperm concentrations in men with secondary infertility, whereas no association was found in men with primary infertility. It may be beneficial for clinicians to advise male patients with secondary infertility who are seeking fertility treatment to avoid heavy alcohol consumption.

Frequency of sperm cells with fragmented DNA in males infected with Chlamydia trachomatis and Mycoplasma sp, determined with the sperm chromatin dispersion (SCD) test

Routine semen analysis provides limited information about a man’s male reproductive potential and can not always fully explain male infertility. Many male infertilities are caused by sperm DNA defects that routine semen quality analyses fail to detect. In this study, we analyzed the association of sperm DNA fragmentation index (DFI) with the semen routine, sperm morphology, in vitro fertilization embryo transfer (IVF-ET)/intracytoplasmic sperm injection (ICSI). Further, we explored the predictive value of sperm DFI in evaluating male fertility and the outcome of IVF-ET/ICSI. Data on sperm DFI, sperm routine, and sperm morphology were collected from 1462 males with infertility. According to DFI levels, there were 468 cases in group I (DFI ≤ 15%), 518 cases in group II (15% < DFI < 30%), and 476 cases in group III (DFI ≥ 30%). The correlations of sperm DFI with semen routine and malformation rate were analyzed. Seminal plasma malondialdehyde (MDA), and total antioxidant capacity (TAC) were assessed. Sperm DFI, semen routine, and sperm morphology were detected in male patients of 101 pairs of IVF-ET/ICSI infertile couples and subdivided into IVF-I group (DFI ≤ 15%), IVF-II group (15% < DFI < 30%), IVF-III group (DFI ≥ 30%), ICSI-I group (DFI ≤ 15%), ICSI-II group (15% < DFI < 30%) and ICSI-III group (DFI ≥ 30%) according to DFI value. The effect of sperm DFI on the outcome of IVF-ET/ICSI was analyzed. There were significant differences in sperm survival rate, sperm concentration, and PR% between groupIII and group II (P < 0.01). There were significant differences in sperm survival rate, sperm concentration and PR% between group III and group I (P < 0.01). There was no significant difference in semen volume, age, abstinence days, or percentage of normal sperm between the three groups (P > 0.05). DFI was positively correlated with MDA content ( P < 0.01) and negatively correlated with TAC (P < 0.01). Sperm DFI was negatively correlated with sperm survival rate, sperm concentration, and PR% (P < 0.01). There was no correlation with age, abstinence days, semen volume, or percentage of normal-form sperm (r = 0.16, 0.05, 0.04, -0.18, p > 0.05). Compared with IVF-I group, the sperm concentration and PR were decreased in IVF-III group. The sperm malformation rate was higher in IVF-III group than that in IVF-II group. Comparatively, the PR was decreased in ICSI-III group. The sperm malformation rate was higher in ICSI-III group than that of the ICSI-I group (P < 0.05). There were no statistically significant differences in fertilization rate, cleavage rate, embryo rate, and clinical pregnancy rate between IVF group or ICSI group, and between all subgroups (P > 0.05). Sperm DFI is negatively associated with sperm survival rate, sperm concentration, and PR%. Antioxidants can decrease the rate of DNA fragmentation. Sperm DFI has proven to be very valuable in the male fertility evaluation, but its significance as a predictor of pregnancy outcomes following assisted reproductive technology. (ART) requires further investigation.

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

Because the assessment of sperm DNA fragmentation (SDF) plays a key role in male fertility, our study was designed to find the relationships between SDF and standard semen parameters. The receiver operating characteristic (ROC) curve showed that 18% SDF is a prognostic parameter for discriminating between men with normal and abnormal standard semen parameters (n = 667). Men with > 18% SDF had significantly lower quality semen, a higher prevalence of abnormal semen characteristics, and a higher odds ratio for abnormal semen parameters compared to men with ≤ 18% SDF. An ROC analysis provided predictive values for age and semen parameters to distinguish between men with SDF > 18% and men with ≤ 18% SDF. SDF was positively correlated with male age and teratozoospermia index but negatively with sperm concentration, total number of spermatozoa, sperm morphology, progressive motility, and vitality. Our study shows that 18% SDF has a predictive value for distinguishing between men with normal and abnormal semen characteristics. Men with >18% SDF have a higher risk for abnormal semen parameters, while age and obtained semen parameters have a predictive value for SDF. There is a relationship between SDF and conventional sperm characteristics, and thus, SDF can be incorporated into male fertility assessment.

To evaluate the effects of a postsurgical rehabilitation program on employment status 2 years after temporal lobe epilepsy surgery in relation to other predictors. Employment outcome 2 years after temporal lobe resection in a group of 232 adult patients with the offer of a 3-week inpatient rehabilitation program immediately after surgery ("Reha group") was compared to a group of 119 patients who had surgery before such a rehabilitation program existed. One hundred thirty-nine (59.9%) of the Reha group patients attended the rehabilitation program. Further predictors for employment outcome were analyzed using multivariate logistic regression analysis. Preoperatively, the groups did not differ significantly in variables relevant for employment, including employment rate. Two years after surgery, the rate of those not being employed had decreased in the Reha group from 38.4% to 27.6% (p < 0.001, McNemar test), and slightly increased in the control group (37.8-42.0%; n.s.), resulting in a difference of 14.4% in favor of the Reha group (p = 0.008). General unemployment rates during the two observation periods were similar. In addition to the offer of rehabilitation support ("Reha group") and preoperative employment, the following other variables were shown as significant predictors of employment post surgery in multivariate regression analysis: seizure outcome, diagnosis of a personality disorder preoperatively, and age at surgery (all, p < 0.01; Nagelkerkes R(2) = 0.59). Independently from other factors, a 3-week inpatient rehabilitation program after temporal lobe epilepsy surgery seems to improve employment status 2 years after surgery.

The incidence and effect of bacteriospermia and elevated seminal leukocytes on semen parameters

Background/AimThis study aimed to evaluate human sperm acrosomal reaction (AR) using flow cytometry and examine the correlation of AR and acrosomal enzyme activity (AEA) with seven semen parameters: total sperm count, concentration, vitality, motility, morphology, survival rate, and sperm DNA fragmentation index (DFI).Patients and MethodsA retrospective analysis was conducted on semen data from 398 men in the AR group and 526 men in the AEA group. AR was assessed using flow cytometry, while AEA was measured with the solid-phase BAPNA method. Correlations between AR, AEA, and the seven semen parameters were analyzed. Subgroups based on standard reference values were also compared.ResultsAEA demonstrated significant positive correlations with total sperm count, concentration, vitality, motility, morphology, and survival rate, and a negative correlation with DFI (p<0.05). In contrast, AR correlated only with sperm concentration. Subgroup analyses revealed that in the AR group, the lower subgroup correlated with concentration and vitality, while the normal subgroup correlated only with concentration. In the AEA group, the normal subgroup showed significant correlations with all seven parameters, while none were found in the lower subgroup. The normal subgroup exhibited significantly higher total count, concentration, vitality, motility, morphology, and survival rate, and lower DFI (p<0.05). No significant differences were observed between the normal and lower AR subgroups, except for survival rate (p>0.05).ConclusionAEA shows stronger and more consistent correlations with semen parameters compared with AR, suggesting its potential as a reliable marker for evaluating male fertility. Further research is needed to clarify the predictive value of AR in male fertility assessment.