Leukocytes In Semen Research Articles

Impact of high-risk and low-risk human papillomavirus infections on the male genital tract: effects on semen inflammation and sperm quality.

Human Papillomavirus (HPV), a prevalent sexually transmitted infection, comprises high-risk (HR-HPV) and low-risk (LR-HPV) viruses, the former posing a high risk for developing malignancies whereas the latter mainly for benign warts. Despite increasing awareness of HPV's impact on men's health, the influence of HR-HPV and LR-HPV urogenital infections on male fertility potential remains uncertain. This study aimed to investigate whether male urogenital infection with HR- or LR-HPV associates with impaired sperm quality, oxidative stress, and inflammation. A total of 205 male patients attending an urology clinic were enrolled. Semen samples were analyzed for HPV using PCR and genotyped by RFLP. Semen quality was evaluated following WHO guidelines. Semen leukocytes, reactive oxygen species (ROS), and sperm viability were analyzed using flow cytometry. HPV was detected in 19% (39/205) of semen samples. HR-HPV infections were more prevalent, with HPV-16 being the most frequent genotype. Neither HR-HPV nor LR-HPV were associated with significant alterations in routine sperm quality parameters. However, HR-HPV+ individuals showed significantly higher levels of sperm necrosis and exhibited increased proportions of ROS+ spermatozoa compared to LR-HPV+ or control individuals. Furthermore, no significant semen inflammation was detected in patients infected with either HR-HPV or LR-HPV, and unexpectedly reduced semen leukocytes and inflammatory cytokines (IL-6 and IL-1β) were observed in HR-HPV+ patients compared to controls. These observations underscore the importance of comprehensive HPV screening, including genotyping, in urology and fertility clinics to understand the progression of the infection, potential adverse effects on reproductive health, and the oncogenic risks involved.

An Alternative Application of Magnetic-Activated Cell Sorting: CD45 and CD235a Based Purification of Semen and Testicular Tissue Samples.

Magnetic activated cell sorting (MACS) is a well-known sperm selection technique, which is able to remove apoptotic spermatozoa from semen samples using the classic annexinV based method. Leukocytes and erythrocytes in semen samples or in testicular tissue processed for in vitro fertilization (IVF) could exert detrimental effects on sperm. In the current study, we rethought the aforementioned technique and used magnetic microbeads conjugated with anti-CD45/CD235a antibodies to eliminate contaminating leukocytes and erythrocytes from leukocytospermic semen samples and testicular tissue samples gained via testicular sperm extraction (TESE). With this technique, a 15.7- and a 30.8-fold reduction could be achieved in the ratio of leukocytes in semen and in the number of erythrocytes in TESE samples, respectively. Our results show that MACS is a method worth to reconsider, with more potential alternative applications. Investigations to find molecules labeling high-quality sperm population and the development of positive selection procedures based on these might be a direction of future research.

(302) Intertest Reliability of Seminal Round Cells and Leukocytes Without Treatment in Men Presenting for Infertility

Abstract Introduction Pyospermia may be due to infection or inflammation along the male reproductive tract. Semen round cells and leukocytes can have a major impact on the management of infertility yet we have a poor understanding of the natural history of patients with seminal round cells or pyospermia. Objective We aimed to assess the intertest reliability of seminal leukocytes and round cells within serial semen analyses in patients without treatment. Methods We conducted a retrospective review of men presenting to a high-volume fertility center for evaluation who had multiple semen analyses assessed for leukocytes between 2/1/2022 and 1/1/2023. Men with azoospermia or who received treatment for pyospermia including antibiotics or surgery in the interval between the tests were excluded. We recorded round cells, the presence of leukocytes, and days between tests. The presence of leukocytes was detected using QwikCheck™ Test Strips which report a positive result if the concentration of leukocytes was greater or equal to 1 million/ml. A Bland-Altman assessment of agreement was conducted to assess the agreement of the number of round cells between the first and second semen analysis. The limits of agreement (LoA) represent the 95% confidence intervals between which we expect most of the differences between measurements. The number of round cells was log-transformed prior to analysis as they were positively skewed. A Cohen Kappa test was used to assess the agreement between the presence of leukocytes in the first sample and the second sample. Results One hundred patients were included in the analysis. The mean time between tests was 75 days (SD: 68.3 days) ranging from 1 to 285 days. There was a median of 2.95 million round cells in the initial semen analysis and 2.45 million round cells in the follow-up semen analysis. Nineteen patients had positive leukocytes on the initial sample and fifteen on the second sample. Eight patients had persistently positive leukocytes. The assessment of agreement for the round cells in the initial and follow-up semen analysis shows a lower LoA of 22% and an upper LoA of 582% meaning that most measurements of round cells in the follow-up tests are estimated to be between 22% and 582% of the initial measurement. The wide range of LoAs suggests poor agreement between tests. The kappa for the presence of leukocytes in the first and second sample was 0.36 (CI 0.13-0.60) which would be considered a fair agreement. Conclusions Round cells and semen leukocytes are poorly understood but are used to aid in the diagnosis and management of male infertility. The number of round cells and the presence of pyospermia between samples varies greatly in patients even without treatment. Providers should consider these discrepancies prior to initiating treatment of pyospermia in men presenting for infertility. Further study is needed to gain a better understanding of how round cells and semen leukocytes should influence management of men with infertility. Providers should be cautious when using the presence of round cells or pyospermia to influence treatment as there is poor consistency between tests even without pyospermia treatment. Disclosure No.

(378) Shorter Abstinence is Associated with a Decrease in Semen Round Cells but Not in Semen Leukocytes

Abstract Introduction The connection between abstinence times and semen round cells and pyospermia is not well understood. However, frequent ejaculation is a commonly prescribed treatment for pyospermia. Objective We sought to identify if a correlation exists between abstinence periods and round cells and semen leukocytes. Methods We conducted a retrospective review of men presenting to a high-volume fertility center for evaluation who had semen samples assessed for leukocytes between 2/1/2022 and 1/1/2023. Azoospermic men and men with abstinence times over 14 days were excluded. We recorded round cells, the presence of leukocytes, and days of abstinence prior to the sample. The presence of leukocytes was detected using test strips which report a positive result if the concentration of leukocytes was greater or equal to 1 million/ml. The study received IRB approval. Pearson correlations (assumes linear relationship) were used as well as generalized additive models (GAMs), which allow for a non-linear relationship between abstinence and the outcome. For analysis of semen leukocytes, a positive result was coded as 1 and a negative result was coded as 0. Round cells were log-transformed prior to calculating correlation. The estimated relationships from GAMs were plotted graphically and summarized via average marginal slopes. Results A total of 382 semen analyses were included, with a mean abstinence period of 3.67 days (SD: 2.11). The number of round cells ranged from 0.2 to 44.0 million (mean: 4.21 million, SD: 4.68), with 19.1% (n = 73) of samples testing positive for leukocytes. Days of abstinence and round cells are significantly correlated according to a Pearson correlation (r = 0.14, p = 0.006). According to the GAM, with each day increase in abstinence, a 0.52 million increase in round cells is expected (CI 0.25-0.79 million; p<0.05). There is an increase in round cells from 0 to 5 days of abstinence before plateauing. Beyond 10 days there is a large degree of uncertainty, due to a low number of observations. When patients with negative pyospermia tests were excluded, there is no linear correlation (r = 0.11, p = 0.36), however, the GAM appears similar to the original sample (1.2; 0.2, 2.1; p<0.05). Semen leukocytes and abstinence are not significantly correlated according to the Pearson correlation (r = -0.04, p = 0.38). There is little change in the probability of leukocytes being present with respect to abstinence in the GAM (-0.001; CI 2.3-2.1; p = 0.926). There is a similar increase in the degree of uncertainty over 10 days. Conclusions These results demonstrate a statistically significant, yet very minimal linear correlation between round cells and days of abstinence. There is an increasing number of round cells up to 5 days of abstinence before plateauing. However, there was no significant correlation between abstinence time and the presence of leukocytes in the semen in either model. Shorter abstinence times may have a role in reducing semen round cells, but it is unlikely to reduce pyospermia. Providers should be hesitant when suggesting an increased frequency of ejaculation to treat pyospermia. Disclosure No.

The Examination of Peroxidase-Positive Leukocytes in Semen.

Leukocytospermia can lead to decreased spermatozoa motility, increased spermatozoa morphological abnormalities, elevated spermatozoa DNA fragmentation index, impairment of the spermatozoa acrosome function, and even affected embryonic development. It is a common andrological disease in clinical practice and one of the important causes of male infertility. When determining whether male reproductive tract inflammation exists, andrologists often choose to examine round cells or seminal plasma elastase in the semen as a clinical diagnostic basis. However, the examination of round cells is easily influenced by sloughed spermatogenic cells and reproductive tract epithelial cells, which do not contribute to reducing the indiscriminate and unnecessary use of antibiotics. At the same time, the detection process of elastase is relatively complicated, time-consuming, and slow in reporting results, which is not beneficial for early diagnosis and treatment of diseases such as male genital tract infections (MGTIs). We have innovatively applied the examination of peroxidase-positive leukocytes in semen assisted by a computer-assisted semen analysis (CASA) system as a diagnostic criterion for leukocytospermia, successfully solving these problems. This examination only requires the addition of the operating fluid consisting of four reagents into the specimen, and the total reaction time at room temperature can be controlled within 20-30 min. With the subsequent smear and microscopic examination, the concentration of peroxidase-positive leukocytes in semen can be obtained within a total of 60 min, which can be used to diagnose whether the inflammation of the male reproductive tract existed.

Antioxidants in Male Infertility-If We Want to Get This Right We Need to Take the Bull by the Horns: A Pilot Study.

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.

COVID-19 associates with semen inflammation and sperm quality impairment that reverses in the short term after disease recovery.

Introduction: COVID-19 exerts deleterious effects on the respiratory, cardiovascular, gastrointestinal, and central nervous systems, causing more severe disease in men than in women. However, cumulative reported data about the putative consequences on the male reproductive tract and fertility are controversial. Furthermore, the long-term effects of SARS-CoV-2 infection are still uncertain. Methods: In this study, we prospectively evaluated levels of inflammatory cytokines and leukocytes in semen and sperm quality parameters in a cohort of 231 reproductive-aged male patients, unvaccinated, who had recovered from mild or severe COVID-19 and in 62 healthy control individuals. Sperm quality was assessed early (less than 3 months) and long (more than 3 and up to 6months) after having COVID-19. Interestingly, and unlike most reported studies, available extensive background and baseline data on patients' sperm quality allowed performing a more accurate analysis of COVID-19 effects on sperm quality. Results: Significantly higher levels of IL-1β, TNF and IFNγ were detected in semen from patients recently recovered from mild and/or severe COVID-19 with respect to control individuals indicating semen inflammation. Moreover, patients recovered from mild and/or severe COVID-19 showed significantly reduced semen volume, lower total sperm counts, and impaired sperm motility and viability. Interestingly, all observed alterations returned to baseline values after 3 or more months after disease recovery. Discussion: These results indicate that COVID-19 associates with semen inflammation and impaired semen quality early after disease. However, long COVID-19 seems not to include long-term detrimental consequences on male fertility potential since the observed alterations were reversible after 1-2 spermatogenesis cycles. These data constitute compelling evidence allowing a better understanding of COVID-19 associated sequelae, fundamental for semen collection in assisted reproduction.

Does folate deficiency alter sperm parameters in infertile males?

Introduction: There is not enough evidence regarding the influence of folate deficiency in the etiology of idiopathic male infertility. Objectives: The objective of the current study was to examine whether there is an association between serum levels of folate and sperm parameters among infertile males. Patients and Methods: This cross-sectional study was undertaken on a sample of infertile males referring to an infertility treatment clinic in Isfahan province, Iran. Semen parameters, comprising sperm concentration, progressive/total motility, morphology, semen leukocyte, agglutination, and vitality were assessed based on standard protocols. The sperm DNA fragmentation assay (SDFA) and sperm chromatin maturity assay (SCMA) were also assessed by the sperm chromatin dispersion and aniline-blue tests respectively. Results: In general, 70 infertile males including 46 with sufficient serum folate level and 24 with insufficient serum folate level were examined. Subjects with sufficient serum folate level had a significantly higher mean of sperm concentration, vitality, progressive motility, total motility, SDFA, and SCMA (P<0.05). Higher serum folate level was also associated with lower risk of abnormal sperm agglutination (OR: 0.09, 95% CI: 0.01-0.65), and morphology (OR: 0.12, 95% CI: (0.02-0.60) after adjustment for potential confounders. Conclusion: The present study found a significant association between serum folate level and infertility among Iranian infertile males. However, prospective large-scale population-based research is required to confirm the finding.

Diff-Quik: A reliable stain for rapid onsite evaluation and differentiation of round cells in semen

Objectives: Identification of immature germ cells and leukocytes and their differentiation pose difficulty in interpretation. In this study, Diff-Quik (DQ) and Papanicolaou (Pap) are used to differentiate between these two, as they are recommended by the WHO, cost-effective and are easy to use. Materials and Methods: This study attempts to differentiate between immature germ cells and leukocytes in semen; compare the usefulness of two stains –Pap and DQ; evaluate the possible associations between the total sperm count and the round cell count; the total sperm count and the presence of round cells and inflammatory cells; the occupation and the sperm concentration; and the occupation and the round cell count. Results: A total of 60 semen samples were included in the study. Two smears were made for each patient; DQ stain was used to stain the air-dried smears and Pap stain for alcohol-fixed smears. The whole procedure of DQ staining takes only 30 s. The smears were compared. There was good concordance between DQ and Pap stains in the detection of inflammatory cells and immature germ cells (P = 0.000). There was no association between total sperm count and round cell count (r = −0.086 and P = 0.520); and between the occupation and the sperm concentration or the round cell count. Conclusion: There was an inverse relationship between total sperm count and presence of immature germ cells (r = −0.291). There was no association between round cell count and presence of immature germ cells (r = 0.148 and P = 0.269), total sperm count and presence of inflammatory cells, round cell count, and presence of inflammatory cells. DQ stain is easy to use, convenient, and rapid. There is good concordance between DQ and Pap stains in the detection of inflammatory cells and immature germ cells. Total sperm count is independent of the round cell count or the presence of inflammatory cells/immature germ cells. In most of the cases, the round cells were the inflammatory cells. Morphological interpretation can be done even in non-liquefied samples.

Semen Leucocytes Affect Sperm Quality of Infertility Patient

The association between the risk factors for male infertility including smoking, obesity, male age, and leukocyte count with sperm analysis, still shows mixed results. This study aims to determine the association between smoking, obesity, male age, and leukocytes count with sperm quality (sperm concentration, sperm motility, and sperm morphology) of infertility patients in Purwokerto. This study is an observational study with a cross-sectional design conducted in the medical records section of the Bunda Arif Hospital Purwokerto. The sample was taken by total sampling. The bivariate test of smoking and obesity behaviour variables with the results of sperm quality using the Chi-Square test and Fisher's exact test. Male age variables used the Kruskal-Wallis test and the Spearman correlation test. Variable leukocytes count using the Spearman test. The results showed no association between male age, obesity and smoking behaviour with sperm quality, sperm concentration, sperm motility, and sperm morphology (p>0.05). There is a significant association (p<0,05) between leukocyte semen count and spermatozoa concentration, sperm motility, and morphology of spermatozoa. It is concluded that there was a significant association between leukocyte count and sperm quality in infertility patients in Purwokerto. Research needs to be continued by examining the relationship between leukocytopenia and sperm DNA damage by looking at sperm DNA fragmentation.

Detection of Trichomonas vaginalis Infection in Chronic Prostatitis/Chronic Pelvic Pain Syndrome Patients by Rapid Immunochromatographic Test.

This study aims to evaluate associations between the immunochromatographic rapid test technique and Trichomonas vaginalis (TV) infection in patients with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) in Taiwan. All patients received post-prostate massage urine (VB3) Trichomonas rapid tests. The demographic characteristics and urogenital symptoms of CP/CPPS were recorded. Routine urinalysis of VB3 was also performed, and laboratory examination results of semen were recorded if available. A total of 29 patients with TV infection and 109 without TV infection were enrolled, which reflected that the prevalence in patients with TV infection was approximately 21%. Patients with TV infection displayed a significantly higher frequency of suprapubic/lower abdominal pain (p = 0.034), semen leukocyte > 5/high-power field (HPF) (p = 0.020), and an inflammatory type (category IIIA) (p = 0.005) than patients without TV infection. A higher prevalence of TV infection was found in patients with category IIIA (47.37%). No significant difference was found in the symptom duration and other clinical symptoms. In conclusion, the high prevalence of TV infection was revealed in CP/CPPS patients using the VB3 rapid Trichomonas test, especially in CP/CPPS patients with category IIIA. Thus, rapid TV testing might be vital for CP/CPPS patients in the hospital.

O-001 Semen impairment and occurrence of SARS-CoV-2 virus in semen after recovery from COVID-19

BackgroundThe presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in human semen and its role in virus contagion and semen quality after recovery from coronavirus disease 2019 (COVID-19) is still unclear. Recent reports evidence that, after SARS-CoV-2 infection, male reproductive function and semen quality may be damaged.AimTo evaluate the semen parameters and inflammation of sexually active men following recovery from SARS-CoV-2 infection at 1 month and 3 months follow-up after the second negative nasopharyngeal swab.Materials and methodsA prospective cross-sectional study on sexually active men recovered from SARS-CoV-2 infection was performed.For previously hospitalized COVID-19 patients, data on serum inflammatory markers were retrospectively collected.One month after the second SARS-CoV-2 negative nasopharyngeal swab and 3 months later, four biological fluid samples, namely saliva, pre-ejaculation urine, semen, and post-ejaculation urine, were collected. The occurrence of SARS-CoV-2 RNA in the specimen was evaluated in all the biological fluids collected by RT-PCR. Female partners were retested if any specimen was found to be SARS-CoV-2 positive.Semen parameters were evaluated according to the World Health Organization manual edition V. Furthermore, semen inflammation was assessed by quantification of semen leukocytes and interleukin-8 (IL-8) levels and evaluation of a panel of sperm cytokine levels by a two-step ELISA method.ResultsA total of 43 men were enrolled in the study. Three patients (7%) tested positive for at least one sample (one saliva; one pre-ejaculation urine; one semen and one post-ejaculation urine), so the next day new nasopharyngeal swabs were collected. The results from these 3 patients and their partners were all negative for SARS-CoV-2.At 1-month follow-up, 25% of the men with recent SARS-Cov-2 infections and proven healing were oligo-cryptoazoospermic, despite the absence of virus RNA in semen. Of the 11 men with semen impairment, 8 were azoospermic and 3 were oligospermic.Serum inflammatory markers (procalcitonin and C-reactive protein) were analyzed in previously hospitalized patients both at admission and at peak of infection. Levels at admission were statistically significantly higher in patients resulting in crypto-azoospermic with respect to those resulting in normozoospermic (p = 0.05; p = 0.03 and p = 0.02, respectively) after healing.Oligo-crypto-azoospermia was significantly related to COVID-19 severity (P < 0.001).A total of 33 patients (76.7%) showed pathological levels of IL-8 in semen.Interleukin-1β and tumor necrosis factor-α levels were significantly negatively related to sperm total number and concentration, whereas interleukin-4 was correlated with sperm motility.At 3-months follow-up, 8/10 men with semen impairment showed an overall increase of semen parameters compared to levels assessed after 1 month. Of the 4 crypto-/azoo-spermic men 1 month after healing, 2 resulted oligozoospermic, 1 normozoospermic and only 1 remained azoospermic. Two of the 3 oligozoospermic men turned normozoozpermic. Semen cytokine levels remained elevated after 3 months, except for IL-6.Discussion and conclusionSARS-CoV-2 can be detected in saliva, urine, and semen in a small percentage of men who recovered from COVID-19. 25% of men who recovered from COVID-19 demonstrated oligo-crypto-azoospermia. Negative correlations between interleukin-1β and tumor necrosis factor-α and sperm number and the overall high levels of semen cytokines indicate a potential detrimental role of SARS-CoV-2 driven inflammation on spermatogenesis.An overall tendency to an improvement of semen parameters was found although a genital tract inflammatory condition appears to persist at least 3 months after COVID-19 recovery.Despite the low number of enrolled patients may limit the statistical power of study and the fact that the previous semen quality of these men was unknown, our results indicate that male of reproductive age recovering from COVID-19 deserve accurate follow-up for their fertility status.

Bariatric Surgery Does Not Improve Semen Quality: Evidence from a Meta-analysis.

The meta-analysis aimed to explore the possible relationship between bariatric surgery and semen quality. PubMed, EMBASE, and CENTRAL were searched from database inception through October 28, 2021. Articles were eligible for inclusion if they evaluated the impact pre- and post-bariatric surgery on semen parameters. A total of 9 studies with 218 patients were found. The mean preoperative age distribution of the patients included centralized from 18 to 50 years, and the mean pre-op BMI ranged from 36.7 to 70.5 kg/m2. The follow-up period ranged from 6 to 24 months. The results revealed that bariatric surgery had no significant effect on sperm volume, concentration, total count, morphology, total motility, progressive motility, viability, semen pH, and semen leukocytes. Bariatric surgery does not improve semen quality in obese males.

Role of Infection and Leukocytes in Male Infertility.

Male infertility is considered as a multifactorial complex reproductive illness, and male urogenital infection and inflammation are crucial etiologies contributing up to 35% of all cases. Mostly triggered by sexually transmitted diseases and uropathogens, chronic manifestation of such infection may cause irreversible infertility in the male. Male urogenital infection involves bacterial, viral, protozoal, and fungal infections many of which remain asymptomatic most of the time and are passed to the sexual partner leading to fertilization failure, pregnancy loss, and even development of illness in the offspring. The abundance of leukocytes in semen can be used as an indicator of urogenital infection. Its contribution in male infertility can be as high as 30% and the clinical condition is referred to as leukocytospermia. Seminal bacterial load together with increased leukocytes contribute to the impairment of male fertility parameters such as, sperm motility, DNA integrity, acrosome reaction, and damage sperm molecular structure. Pathophysiology of bacteriospermia-induced impairment of male infertility is probably mediated by the involvement of bacterial pathogens in the intrinsic apoptotic pathway resulting in sperm death, whereas that of seminal leukocytes operates through excessive generation of ROS. Although the application of antibiotics forms the frontline therapeutic approach, the growing resistance to antibiotics poses a concern in the management of microbes-induced male urogenital infection. Complementary and alternative medicine may offer additional management options in combating such infections. On the other hand, both broad spectrum antibiotics and antioxidant therapy have showed promising results in the management of infertile men with leukocytospermia. Use of herbal medicine may also play a promising role in the management of such patients. However, recent molecular biology techniques have noted the association of elevated levels of IL-8 with both the Chlamydial infection of the male urogenital tract as well as the clinical condition of leukocytospermia. On the basis of such common pathogenesis, further research involving advanced molecular techniques may pave the way towards the development of better diagnostic tools in the clinical management of male urogenital infection and leukocytospermia.

Leukocytospermia is related to total motile sperm counts in infertile men at Sadewa Maternal and Child Hospital, Yogyakarta, Indonesia

Background: Leukocytospermia is defined as the presence of leukocytes in semen at the concentration of ? 1 x 106/mL according to World Health Organization (WHO) manual procedure. Leukocytes are the source of reactive oxygen species (ROS), which may lead to oxidative stress conditions and sperm damage. This study aims to evaluate the association between leukocytospermia, poor semen quality, and infertility at Sadewa Maternal and Child Hospital, Yogyakarta, IndonesiaMethods: This study was based on secondary data of semen analyses of males attending IVF clinics at Sadewa Maternal and Child Hospital, Yogyakarta, Indonesia, from May 2015 to November 2020. Variables assessed in this study were semen volume, sperm concentration, progressive motility, and Total Motile Sperm Count (TMSC). Data were analyzed using SPSS version 23 for Windows.Results: The median sperm concentration of the leukocytospermia group was higher than that of the non-leukocytospermia group. There is a relationship between the number of leukocytes in semen and TMSC, with the higher TMSC median in the leukocytospermia group.Conclusion: This is the first study to report a significant association between leukocytospermia and total motile sperm count (TMSC). The association is not expected with leukocytospermia patients who have better TMSC value.

Association of sexually transmitted infection with semen quality in men from couples with primary and secondary infertility

This study aims to compare the prevalence of sexually transmitted infections (STIs) with semen quality in men from couples with primary and secondary infertility. Semen samples were collected from 133 men who requested fertility evaluation. Seminal tract infection with Ureaplasma spp. (UU), Mycoplasma hominis (MH), Mycoplasma genitalium (MG), Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and herpes simplex virus-2 (HSV-2) was assessed by PCR-based diagnostic assays. Among all patients, the prevalence of STIs was higher in men from couples with primary infertility than that in men from couples with secondary infertility (39.7% vs 21.7%, P = 0.03). The prevalence of UU was 28.8% and 13.3% in men from couples with primary and secondary infertility, respectively. Men from couples with primary infertility were more likely to be positive for UU than men from couples with secondary infertility (P = 0.04). Regarding the UU subtype, the prevalence of Ureaplasma urealyticum (Uuu) and Ureaplasma parvum (Uup; including Uup1, Uup3, Uup6, and Uup14) did not differ between the two groups. No associations between the prevalence rates of MH, MG, and CT were found in men from either infertility group. A lower sperm concentration was associated with STI pathogen positivity in men with primary infertility according to the crude model (P = 0.04). The crude and adjusted models showed that semen volume (both P = 0.03) and semen leukocyte count (both P = 0.02) were independently associated with secondary infertility. These findings suggest the importance of classifying the type of infertility during routine diagnosis of seminal tract infections.

Bacterial communities in bovine ejaculates and their impact on the semen quality

ABSTRACT Although bacterial contamination of ejaculates may cause difficulties in cattle reproduction, standard protocols for a routine microbiological analysis of bovine semen are still missing. Understanding of the mechanisms of bacterial damage to spermatozoa may contribute to the prevention and management of bacteriospermia in the future. Therefore, this study was designed to investigate bacterial profiles of fresh bovine ejaculates (n = 30), while at the same time we focused on assessing the relationships between bacteriospermia and selected sperm quality parameters as well as an array of oxidative stress and inflammatory markers. The samples were divided into three quality groups according to the sperm motility: Excellent (EX) – over 90% > Good (GO) – between 89% and 80% > Moderate (MO) – under 80%. The results showed a significant increase in reactive oxygen species (ROS) generation in the GO group when compared to the EX group. In the MO group, a deterioration of almost all quality parameters was observed when compared to the EX group. In particular, sperm motility, mitochondrial membrane potential, ROS production and IL-6 concentration exhibited a significant decline. Pearson correlation analysis revealed positive associations among bacterial load and the presence of leukocytes in semen (r = 0.965), malondialdehyde concentration (r = 0.816) and DNA fragmentation (r = 0.784). MALDI-TOF MS Biotyper analysis showed a prevalence of the Staphylococcus genus. The quantification of bacterial colonies revealed a significantly increased (P < 0.01) bacterial load in the MO group when compared with the EX as well as the GO group. Overall, our results suggest that sperm quality may be affected by both, bacterial composition, and bacterial load. It appears that an increased presence of bacterial species triggers the immune response, causes oxidative stress, and thereby contributes to sperm structural alterations while diminishing their fertilization ability. Abbreviations: EX: Excellent; GO: Good; MO: Moderate; MOT: Motility; ROS: Reactive Oxygen Species; MMP: Mitochondrial Membrane Potential; IL-1: Interleukin 1; IL-6: Interleukin 6; IL-8: Interleukin 8; IL–12: Interleukin 12; CRP: C-reactive protein; DNA: Deoxyribonucleic acid; MALDI-TOF MS: Matrix-assisted Laser Desorption/Ionization–Time of Flight Mass Spectrometry; LPO: Lipid peroxidation; CFU: Colony-forming unitsMDA: Malondialdehyde; CASA: Computer-assisted Sperm Analysis; WS: Working solution; RIPA: Radio–immunoprecipitation assay; TBARS: Thiobarbituric Acid Reactive Substances; BHB: D–β–hydroxybutyrate.

Effects of Selenium Supplementation on Sperm Parameters and DNA-Fragmentation Rate in Patients with Chronic Autoimmune Thyroiditis.

Background: Selenium (Se) is an essential component of selenoenzymes, which have catalytic and antioxidant functions. A low Se status has been reported in patients with chronic autoimmune thyroiditis (AT) who benefit from Se supplementation. The role of Se in male reproduction is still a matter of debate. Although Se and selenoenzymes ensure sperm viability and protect against increased oxidative stress, only a few studies have assessed the effects of the administration of Se alone on sperm parameters, providing contrasting results. Aim: The aim of this study was to assess the effects of oral Se supplementation on conventional sperm parameters and DNA fragmentation (SDF) in patients with AT of reproductive age with normal thyroid function. Patients and Methods: Only patients with AT and normal thyroid function were selected for this study. All included patients underwent oral Se supplementation at the dose of 83 µg once daily (Syrel®, IBSA) for six months. Sperm conventional parameters, SDF, and thyroid function were assessed before and at the end of the treatment. Results: Twenty AT patients with normal weight were enrolled. After Se supplementation, they showed a higher sperm concentration, a higher percentage of sperm with progressive motility, and a higher percentage with normal morphology. They also had lower semen leukocyte concentration, and a lower percentage of spermatozoa with DNA fragmentation compared with pre-treatment values. Free-thyroxine serum levels increased significantly, whereas free triiodothyronine showed an upward trend. The thyroid-stimulating hormone did not change significantly. Conclusion: Se supplementation may represent a possible non-hormonal therapeutic choice for the treatment of male infertility, although further studies are needed to confirm this evidence. The possible thyroid hormone dependency of these findings needs to be clarified.

Semen impairment and occurrence of SARS-CoV-2 virus in semen after recovery from COVID-19.

STUDY QUESTIONHow is the semen quality of sexually active men following recovery from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection?SUMMARY ANSWERTwenty-five percent of the men with recent SARS-Cov-2 infections and proven healing were oligo-crypto-azoospermic, despite the absence of virus RNA in semen.WHAT IS KNOWN ALREADYThe presence of SARS-CoV-2 in human semen and its role in virus contagion and semen quality after recovery from coronavirus disease 2019 (COVID-19) is still unclear. So far, studies evaluating semen quality and the occurrence of SARS-CoV-2 in semen of infected or proven recovered men are scarce and included a limited number of participants.STUDY DESIGN, SIZE, DURATIONA prospective cross-sectional study on 43 sexually active men who were known to have recovered from SARS-CoV2 was performed. Four biological fluid samples, namely saliva, pre-ejaculation urine, semen, and post-ejaculation urine, were tested for the SARS-CoV-2 genome. Female partners were retested if any specimen was found to be SARS-CoV-2 positive. Routine semen analysis and quantification of semen leukocytes and interleukin-8 (IL-8) levels were performed.PARTICIPANTS/MATERIALS, SETTING, METHODSQuestionnaires including International Index of Erectile Function and Male Sexual Health Questionnaire Short Form were administered to all subjects. The occurrence of virus RNA was evaluated in all the biological fluids collected by RT-PCR. Semen parameters were evaluated according to the World Health Organization manual edition V. Semen IL-8 levels were evaluated by a two-step ELISA method.MAIN RESULTS AND THE ROLE OF CHANCEAfter recovery from COVID-19, 25% of the men studied were oligo-crypto-azoospermic. Of the 11 men with semen impairment, 8 were azoospermic and 3 were oligospermic. A total of 33 patients (76.7%) showed pathological levels of IL-8 in semen. Oligo-crypto-azoospermia was significantly related to COVID-19 severity (P < 0.001). Three patients (7%) tested positive for at least one sample (one saliva; one pre-ejaculation urine; one semen and one post-ejaculation urine), so the next day new nasopharyngeal swabs were collected. The results from these three patients and their partners were all negative for SARS-CoV-2.LIMITATIONS, REASONS FOR CAUTIONAlthough crypto-azoospermia was found in a high percentage of men who had recovered from COVID-19, clearly exceeding the percentage found in the general population, the previous semen quality of these men was unknown nor is it known whether a recovery of testicular function was occurring. The low number of enrolled patients may limit the statistical power of study.WIDER IMPLICATIONS OF THE FINDINGSSARS-CoV-2 can be detected in saliva, urine, and semen in a small percentage of men who recovered from COVID-19. One-quarter of men who recovered from COVID-19 demonstrated oligo-crypto-azoospermia indicating that an assessment of semen quality should be recommended for men of reproductive age who are affected by COVID-19.STUDY FUNDING/COMPETING INTEREST(S)None.TRIAL REGISTRATION NUMBERN/A.

Application of leukocyte subsets and sperm DNA fragment rate in infertile men with asymptomatic infection of genital tract.

About 15% of male infertility is due to genital tract infection and inflammation, some of them have no clinical symptoms, but manifested as leukocytospermia (LCS). Leukopenia will lead to functional impairment of male sperm and integrity damage of sperm morphology. A large amount of reactive oxygen species (ROS) produced by leukocytes can damage sperm nuclear DNA. The aim of this study was to investigate the correlation between leukocyte subsets and sperm DNA fragmentation rate in semen of infertile men with asymptomatic infection of genital tract. One hundred and eight cases of infertile men were enrolled, who were admitted to our hospital from May 2016 to September 2018, and all had genital tract infections. After routine sperm analysis, realtime PCR was performed for detecting the infection of chlamydia trachoma (CT) and Ureaplasma urealyticum (UU). Furthermore, total leukocyte count in semen was evaluated by detection of CD45 molecules using immunocytochemistry. Flow cytometry was used for subset analysis, monocyte/macrophages were evaluated by CD14, and activated macrophages were evaluated by HLA-DR antigen. Sperm DNA fragmentation index (DFI) were evaluated by SCD method and 8-hydroxydeoxyguanosine (8-OHdG) expression were evaluated by chromatin diffusion method and TUNEL method; the correlation analysis was conducted between semen leukocyte subsets, sperm DNA fragment rate and conventional semen parameters. There was a significant correlation among the concentrations of cells expressing HLA-DR antigen and those expressing CD14 (P<0.01), but the concentrations of differential leukocyte subsets all had no significant correlation with sperm DFI, the percentage of 8-OHdG-expressing cells and routine semen parameters. The percentage of 8-OHdG-expressing sperm was positively correlated with the percentage of sperm fragments (r=0.42, P<0.01), and negatively correlated with sperm concentration (r=-0.32, P<0.01). After adjustment for possible confounders including age, abstinence time and smoking, the percentage of 8-OHdG-expressing sperms independently associated with sperm concentration (β=-0.30; P=0.006) and DFI (β=0.180, P=0.06). Among infertile men with genital tract infection, the sperm DFI is associated with decreased semen quality and not the concentration of differential leukocyte subsets.