Spontaneous Alterations in Semen Parameters are Associated with Age, Accesssory Gland Function and the FSHB c.-211G>T Variant – Maria Schubert

Publications on oligozoospermia states that subject variations are observable around certain homeostatic setting points such as age, accessory gland function, testicular function, related genetics, and FSH (follicle-stimulating hormone). WHO suggests two semen analysis, 60-90 days apart, covering two spermatogenic cycles in in fertility patients. This study aimed to elucidate the subgroup of patients in which these variations are likely to occur and the semen parameters that are likely to be affected without clinical intervention. Factors to be stratified included testicular volume, markers of accessory gland function, hormones, and genetic setting points. Hormones that mostly affect spermtogenesis are FSH and SNP (single-nucleotide polymorphism). Earlier studies have shown that FSHB gene c.-211G>T impairs transcription factor binding, leading to reduced FSH production and secretion. Comparing patients carrying G and T allele, it was observed that patients with T allele had significantly reduced sperm count, testicular volume and serum FSH. The current study was a retrospective, longitudinal study that included men for fertility workup with at least two semen analyses 70-90 days apart. Idiopathic/unexplained male infertility with sperm concentration >1 mill/ml, ejaculate volume >0.8 ml, no signs of infection, and full data set with semen parameters, ultrasound and genotyping were included. A total of 3456 patients were selected and divided into 3 groups: group A, severe oligozoospermia (n=397), group B, mild-moderate oligozoospermia (n=708), and group C, normal concentration (n=2351). Group A patients had more patients with T allele genotype and higher FSH. For statics, categorical parameters were selected which were considered clinically relevant if changes in ejaculate volume and FSH were ≥1.8 ml and ≥1.5IU/l, respectively, between first and second visit. In group B, change in sperm level was due to change in FSH level that mostly occurs spontaneously in men carrying the wildtype GG. Total sperm count and sperm concentration may change up to 10-15% over time either way. This impact comes from ejaculate volume which itself it impacted by abstinence time, younger age and normal testicular volume. Sperm count and concentration is also impacted by FSH which itself is dependent on the genotype. The patient subgroup most affected by the variations are younger men with normal testicular volume and low to almost normal sperm concentration. Ejaculate volume due to accessory gland function impairment can be changed therapeutically by reducing inflammatory stress. FSH level can be increased by FSH treatment, which could be the basis for future clinical trials.

Sperm Motility has Declined from 2017-2022 among Candidate Sperm Donors in Denmark – Robert Montgomerie

Many studies over the last 10 years have suggested that sperm count has declined since the 1970s. However, more recent studies have identified potential problems with previous studies in terms of inadequate statistical analysis, changes in semen analysis protocols in different studies and data from different male volunteers (recent fathers, healthy volunteers, etc.). To address that literature confusion, this present study was set up to analyze semen characteristics of sperm donor applicants and accepted sperm donors from four clinics in Denmark. At each clinic, exact same CASA (computer assisted sperm analyses) software and semen handling protocols were followed. The two groups of this study were: (1) applicants (n=6758 men) that donated one semen sample each and (2) subset of those participants who passed minimum required criteria, accepted as sperm donors (n=1839) and each of them donated several semen samples over the course of the study. Fewer donors were accepted from age group <25 years. For analyses, controls applied were male age, mean monthly temperature, cities and multiple samples from donors. Semen traits measured were ejaculate volume, sperm concentration per mL, total sperm count, motile sperm concentration and total motile sperm count (TMSC) classified as grade a (rapidly progressive) and grade b (slowly progressive). Results showed very less change in ejaculate volume and sperm concentration (<5%) over the 5 years of study period. There was a steep decline in sperm motility (>30%) both in TMSC and motile sperm concentration. Both grade a and b sperm concentration declined from the year 2019 to 2022. On analyzing individual males, 80% men who donated semen during the study had no sperm count decline at all. Motile sperm concentration of a few donors increased significantly, while twice as many decreased significantly (about 10% of all men). Overall, a remarkably steep decline was seen in both grade a and b TMSC, supporting the findings of other studies, but this decline was only in 10% of men. This steep decline between 2019-2022 perfectly matches the COVID-19 pandemic period. The possible reasons of the decline could be changes in male behavior during the pandemic, such as increased sedentary levels, increased drug/alcohol use, or it could be certain general societal changes unrelated to the pandemic. For future studies, more data are needed including more information from sperm donors to identify possible causes of the decline and decide on intervention to improve sperm motility based on individuals.

A Novel Diagnostic Test Identifies Patients Suffering from Loss of CatSper Function – Samuel Young

Outcomes of semen analysis of infertile men have shown 30% of patients to be normozoospermic, displaying unexplained sperm dysfunction. It is difficult to decide the therapy for these patients. As a sperm tries to penetrate an egg, it encounters progesterone hormone secreted by cumulus cells surrounding the egg. This gives hyperactive motility to the sperm, providing it the force to penetrate the egg coat and ultimately fertilize the egg. The target of progesterone in sperm is CatSper channel, which is a calcium channel found exclusively in flagellum of sperm and it modulates the swimming behavior. CatSper is essential for fertilization and without it, men are infertile. The aim of the study was to identify CatSper dysfunction in infertile patients. To do that, CatSper test was created in which ejaculate is added into a buffer solution that makes sperm immotile. This buffer only works when CatSper is functional. Thus, when CatSper inhibitor is added to the sperm, it continues to swim in the buffer. For evaluation, 2286 patients were screened, and 8 patients with loss of CatSper function were identified. These patients were genetically analyzed and all had homozygous deletion on chromosome 15 which included the CatSper 2 gene that made the whole channel dysfunctional. 7 of these 8 patients were normozoospermic and confirming this hypotheses explains the cause of unexplained infertility in them. Sperm of these patients failed to get hyperactivated after exposure to progesterone, thus confirming infertility. There was total fertilization failure when these sperms were subjected to in vitro fertilization and only ICSI (intracytoplasmic sperm injection) worked for these patients.

In summary, infertility in normozoospermic patients is not uncommon. In this study, prevalence of CatSper dysfunction was 1.2% in normozoospermic infertile men. The CatSper test is a simple technique to provide evidence-based diagnosis of CatSper dysfunction. Apart from ICSI, other fertilization therapies would fail in these cases.

Impact of Atypical Bacterial Semen Infections on Spermocytogram Parameters and Spermatozoa DNA Fragmentation and Denaturation – Dr Hatem Bahri

Numerous bacteria have been isolated from semen and genital tract of infertile men that can impact spermatogenesis process at different levels and is associated with poor sperm function, leading to infertility. Some bacterial endotoxins, such as lipopolysaccharide and glycoprotein detected in semen have shown to affect semen quality by triggering a local inflammatory reaction. This further activates leukocytes and mediators such as cytokines and reactive oxygen species (ROS), which play significant roles in sperm DNA fragmentation (SDF) and denaturation (SDD). The study aimed to evaluate the impact of atypical bacterial semen infections on spermocytogram parameters, SDF and SDD levels by analyzing 10-year results conduced from January 2013 to December 2022. It included 10,386 men between age 19-73 years, divided into two groups, group 1 was infection positive group (n=613) and group 2 was control (n=9773). Group 1 was further divided into 6 subgroups based on nature of the bacteria (5 with mono-atypical and 6^th with all poly-atyical bacterial infection). For each patient, sperm culture, spermocytogram, SDF and SDD test were carried out to identify a correlation. Study results showed significantly lower sperm concentration, motility and typical morphology in the event of an infection (group 1). Leukocyte concentration and SDD were significantly higher in group 1. Similar results were observed with 3/5 of the monobacterial semen infected subgroups (MH, UU and UP), while remaining two (MG and CT) showed no difference except for higher levels of SDD in CT-subgroup. SDD was positively correlated to the presence of MH and UU, leukocyte concentration was positively correlated to UU, MH, CT and UP. Sperm concentration, motility and morphology had negative correlations with UU, MH and UP. SDF showed no significant variation between studied groups. These results are in accordance with the 2013 study of Lee et al. showing significantly lower semen motility in UU-positive cases and low total motility in MH-positive cases. SDD showed positive correlation with semen infection with MH and UU. Sperm DNA damage is linked to abortive apoptosis during meiosis I, problem in chromatin condensation during spermiogenesis, oxidative stress and ROS generation.

To conclude, North African men’ sperm parameters and DNA integrity could be affected by certain bacterial infections, the mechanisms of which are not yet established. Positive sperm culture rate in this study was lower (6%) than other similar worldwide studies. It could be due to the difference of climate, ethnicity and lifestyle. Meta-analysis and more prospective randomized controlled trials are necessary to confirm the results of this study.

Abbreviations- CT: Chlamydia trachomatis, MG: Mycoplasma genitalium, MH: Mycoplasma hominis, UU: Ureaplasma urealyticum, UP: Ureaplasma parvum 

Decreased Sperm Motility Associated with Bacterial Vaginosis Modeled in a Human Cervix Chip – Ola Gutzeit

Microbiome play an important role in women health. However, the causation of dysbiotic condition to infertility is still under evaluation. Studies have shown higher biodiversity of bacterial species in cervical samples of infertile women, reduced fecundability in women with bacterial vaginosis (BV), and higher proportion of lactobacilli-impaired microbiota in women with unexplained infertility. A healthy microbiome of lower reproductive tract has lower amount of bacteria with lactobacilli being dominant which protects epithelium, produce mucins and reduce inflammatory state. In dysbiotic condition, there is higher variety of anaerobic bacteria which can produce biofilm and increase inflammation. The most abundant dysbiotic condition is BV which affects >25% of women of reproductive age and has stronger associated risk of preterm birth and acquiring sexually transmitted diseases like HIV (human immunodeficiency virus). Despite treatment, the recurrence rate is >60%. To mimic a lower reproductive tract with a good model, it is important to know what to mimic in terms of microbiome. It should have a good epithelial layer and good mucus production. An organ-on-a-chip platform has a microfluidic chain and porous membrane to separate two channels. The porous membrane is covered with extracellular matrix print with the ability to see different cell types on each side of the membrane. There are vacuum channels on the sides that can cause contraction of the chip. On the vagina chip, the top is the vaginal epithelial layer and bottom is vaginal fibroblast layer. This can mimic the physiological markers of the vaginal epithelium. Similarly on the cervix chip, the top and bottom are cervical epithelial and fibroblast layers, respectively, that mimics the markers in vivo. The aim was to evaluate the effects of BV on sperm function using the chip. Clinically, decrease in sperm motility was observed in Gardnerella vaginalis (GV) and BV sample cultures. However, it does not mimic in vivo activity of the sperm. To evaluate this, chips were cultured with good (lactobacilli consortia, LC) and bad bacteria (GV) in this study as it faithfully model mucosa-microbiome interactions. Cytokine levels increased with GV in both cervix and vagina chip. The epithelial barrier was maintained in LC but was disrupted with GV. The number of bacteria under good condition was maintained at same level on chip in 72 hours, but there was a significant increase in amount under dysbiotic condition. Mucus production was more with LC, whereas, it was thinner and patchy with GV. Sperm was introduced to the cultured chips after 48 hours. After 24 hours, significantly lower motility of sperm was observed on dysbiotic chip. BV consortia decreased sperm migration on cervix chip compared to healthy consortia. The ability of sperm to bind to hyaluronic acid was significantly reduced with the presence of dysbiotic condition.

To conclude, this study suggested that infertility related to BV may cause sperm dysfunction. This model can be used to identify dysbiotic conditions in the female reproductive tract that are unfavorable to sperm migration.

What is the Recovery Time for Sperm Parameters in Men who have Suffered a Mild COVID-19 Infection? - Núñez-Calonge R

Several reports indicated that approximately 58% of SARS-CoV-2 infected patients are male, suggesting male sex as one of the risk factors for the disease. Testosterone and androgen receptors, which are coregulators of ACE2 (Angiotensin-converting enzyme 2) and TMPRSS2 (Transmembrane protease, serine 2) expression, probably help in the internalization of SARS-CoV-2 in the male reproductive organs. However, COVID-19 has yet to be proven to alter male reproductive function. Previous studies evaluating the effect of COVID-19 infection on semen parameters have insufficient data and are inconclusive regarding the long-term impact. The purpose of this study was to assess the effect of mild/asymptomatic SARS-CoV-2 infection on semen parameters in a cohort of men recovered from COVID-19 and the time required for the seminal parameters to be restored to the pre-COVID values. This prospective, multicentre study in Spain included 45 patients (aged 18-46 yrs) for infertility study with semen analysis done before and after COVID-19. The diagnosis of mild COVID-19 was confirmed in these patients, and at the time of disease, no patient was vaccinated. Analyses included volume, sperm count, total sperm count, motility, morphology and vitality. No significant correlation was found between the age of the participants and semen parameters analyzed. Sperm parameters before and after having COVID-19 were compared, and except for vitality, there was a clear correlation between the values for all parameters before and after the infection. Values of all parameters decreased significantly after infection. Morphology was not affected. Vitality decreased by 10%, total sperm count decreased by 39%. Half of the individuals obtained a total count in the second sample less than 57% of the initial one. Pair of values between the time elapsed from infection to the subsequent semen analysis showed no significant change, indicating that the effect of COVID-19 on semen remained unchanged over time, within the range of this study. There was no difference in any parameters between early group (<100 days of infection) and late group (>100 days of infection).

In conclusion, the effect of COVID-19 infection on the concentration and sperm motility is clearly confirmed and is maintained over time, at least beyond 100 days. The present study revealed the long-term impacts of SARS-CoV-2 on various parameters of sperm quality even with a mild infection. Clinicians should be aware of the damaging effects of SARS-CoV-2 virus on male fertility. Further efforts should be made to study long-term effects of COVID-19 infection on male reproduction.