Abnormal Sperm Morphology Research Articles

Alterations in testis histology, reproductive hormones and abnormal sperm morphology in mice treated with polyherbal antimalarials

Alterations in testis histology, reproductive hormones and abnormal sperm morphology in mice treated with polyherbal antimalarials

The effect of temperature and storage time on DNA integrity after freeze-drying sperm from individuals with normozoospermia.

This study evaluated the effects of temperature and storage time on the quality and DNA integrity of freeze-dried sperm from individuals with normozoospermia. Normal sperm samples from 15 men aged 24 to 40 years were studied. Each sample was divided into six groups: fresh, freezing (frozen in liquid nitrogen), freeze-dried then preserved at room temperature for 1 month (FD-1m-RT), freeze-dried then preserved at room temperature for 2 months (FD-2m-RT), freeze-dried then preserved at 4 °C for 1 month (FD-1m-4 °C), and freeze-dried then preserved at 4 °C for 2 months (FD-2m-4 °C). The morphology, progressive motility, vitality, and DNA integrity of the sperm were evaluated in all groups. In all freeze-dried groups, sperm cells were immotile after rehydration. The freeze-dried groups also showed significantly less sperm vitality than the fresh and frozen groups. Significantly more morphological sperm abnormalities were found in the freeze-dried groups, but freeze-drying did not lead to a significantly higher DNA fragmentation index (DFI). The DFI was significantly higher in the FD-2m-RT group than in the other freeze-dried groups. The freeze-drying method preserved the integrity of sperm DNA. The temperature and duration of storage were also identified as factors that influenced the DFI. Accordingly, more research is needed on ways to improve sperm quality in the freeze-drying process.

Adverse effects of exposure to petrol-generator exhaust fumes on the reproductive hormones, testis and spermatozoa in male dogs

There is evidence that sperm count has progressively declined in men over the recent decades. Exposure to air pollutants including petrol and diesel exhaust have been reported to impair male reproduction although there is little experimental evidence. This study investigated the effects of petrol-generator exhaust fumes (PGEF) on semen, sperm, gonadal structure and hormonal status in the dog. Sixteen adult male Basenji dogs were randomly assigned four to each of 4 groups as follows: an unexposed (Control) group and three groups exposed to graded levels of PGEF for 1, 2 or 3 h per day (hpd), respectively, for 90 days. Serum concentrations of testosterone (T), follicle stimulating hormone (FSH) and luteinizing hormone (LH) were measured on days 0 (baseline), 30, 60 and 90 of the study. At day 90, semen samples were collected for semen and sperm analysis. Testicular and epididymal tissues were subjected to gross, histopathological and histomorphometric evaluation. Graded exposure to PGEF resulted in increased serum concentration of T and decreased concentrations of FSH and LH, increased seminal plasma lipid peroxidation, seminiferous and epididymal tubular degeneration, germ cell depletion, lowered sperm concentration, decreased sperm motility and vitality, and increased sperm abnormal morphology. The close proximity between dogs and humans in exposed environments underscores the importance of these findings to human reproductive health and fertility. The findings suggest that with prolonged exposure, the impairment of reproductive functions will likely play significant roles in the decline in male fertility.

Dnali1 is required for sperm motility and male fertility in mice

BackgroundThe sperm flagellum is an evolutionarily conserved specialized organelle responsible for sperm motility and male fertility. Deleterious mutations in genes involved in the sperm flagellum assembly can often cause sperm motility defects and male infertility. The murine Dnali1 gene encodes a protein that is known to interact with the cytoplasmic dynein heavy chain 1.ResultsA Dnali1-mutated mouse model was generated by inducing a nonsense mutation in the Dnali1 gene. The Dnali1-mutated male mice presented impaired sperm motility and were completely infertile. Although no obviously abnormal sperm morphology was observed in Dnali1-mutated male mice, the ultrastructural structure of sperm flagellum was disrupted, displaying as an asymmetrical distribution of the longitudinal columns (LCs). Notably, infertile Dnali1-mutated male mice were able to obtain offspring via ICSI.ConclusionsOur results uncover a role of DNALI1 in sperm motility and male fertility in mice, and demonstrate that ICSI overcomes Dnali1-associated male infertility, thus providing guidance for the diagnosis and genetic counseling of DNALI1-associated human infertility.

Effect of varicocele surgery on sperm quality

Introduction: A varicocele is a vascular lesion characterised by dilatation and tortuosity of the veins of the pampiniform plexus and is found in approximately one-fifth of the adult male population. It is the most common cause of treatable infertility. Physical examination may or may not show a varicocele which is mostly diagnosed only when patients present with infertility or adolescents during a physical examination, like testicular hypotrophy, impairment in spermatogenesis mainly in the form of low or absent count, decreased sperm motility, and abnormal sperm morphology. Treatment for varicocele includes medical therapies, radiological embolization, and surgical techniques. Medical therapy, including antioxidants and anti-inflammatory agents, has been utilized with variable success. Radiological embolization of varicocele is a new technique in the treatment of varicocele, but it is associated with high recurrences and high cost. Varicoceles are surgically treated either by open or laparoscopic approaches the principal aim being occlusion of the dilated veins of the pampiniform plexus. A definitive conclusion of indication of medical treatment cannot be drawn at present because most published studies have inadequate design and lack controls. Various studies in the past have concluded significant changes in sperm parameters (total count, motility, morphology) post-varicocele repair. However, few other studies suggest no improvement in sperm parameters following surgical varicocele repair and studies that conclude recommendations against repair of varicocele. Thus, conflicting opinions have opened the scope to establish this fact by this study. Aim: To evaluate changes in sperm morphology, sperm count, and sperm motility in a patient with varicocele after varicocele surgery. Materials and methods: The study was conducted in the Department of Surgery, North Delhi Municipal Corporation Medical College & Hindu Rao Hospital, Delhi, India from August 2019 to February 2021. The sample size was calculated to be 50 using Slovin’s formula with a confidence level of 95% with a margin error taken as 5% (p - value 0.05). Sperm morphology, sperm count, and sperm motility were the parameters studied before and after 3 months of surgery. Results: The majority of patients belonged to the age group of 18 to 25 years (42%) followed by 26 to 35 years (40%) (27.78 ± 6.547 years). Left-sided varicocele was more common (54%), followed by bilateral varicocele (42%). Varicocele grade II was most common (58%) followed by grade I (30%) and lastly grade III (12%). Total sperm count improved by 8.88 million per ml after varicocelectomy. Total sperm motility improved by 6.08% after the surgery. Total normal sperm forms improved by 6.44% after varicocele repair. Conclusion: Based on the findings it was concluded that Varicocelectomy results in significant improvement in total sperm count, total sperm motility, and total normal sperm morphology. The best improvement was seen in sperm count followed by total normal sperm forms and lastly total sperm motility.

Effect of white guava (Psidium guajava L.) fruit juice on the quality of lead acetate induced rats (Rattus norvegicus) spermatozoa

This study aims to determine the effect of the administration of white guava (Psidium guajava L.) fruit juice on spermatozoa plasma membrane integrity (PMI), morphological abnormality, viability, and motility of lead acetate induced rats (Rattus norvegicus). Twenty-five male rats were divided into five groups: NC (negative control) group, rats were administered with distilled water twice daily at four-hour intervals; T0 (positive control) group, rats were administered daily with lead acetate 50 mg/kg bw and distilled water four hours later; T1, T2, and T3 groups, rats were administered daily with lead acetate 50 mg/kg bw and 0.5 mL of 25, 50, and 100% white guava fruit juice four hours later. The treatment of the rats was conducted for 14 days, and on day 15, all rats were sacrificed to assess the spermatozoa quality. Data was analyzed using ANOVA followed by Duncan's multiple range test at a confidence level of 95%. The results showed that exposure to lead acetate (T0) caused lower spermatozoa PMI, viability, and motility as well as higher spermatozoa morphological abnormalities (p <0.05) compared to those of the T0 group. Administration of white guava fruit juice starting at a dose of 25% (T1) resulted in higher spermatozoa motility, viability, and PMI as well as lower spermatozoa morphological abnormalities (p <0.05) compared to rats in the T0 group. It could be concluded that white guava fruit juice maintained the spermatozoa quality of lead acetate induced rats.

Effect of white guava (Psidium guajava L.) fruit juice on the quality of lead acetate induced rats (Rattus norvegicus) spermatozoa

This study aims to determine the effect of the administration of white guava (Psidium guajava L.) fruit juice on spermatozoa plasma membrane integrity (PMI), morphological abnormality, viability, and motility of lead acetate induced rats (Rattus norvegicus). Twenty-five male rats were divided into five groups: NC (negative control) group, rats were administered with distilled water twice daily at four-hour intervals; T0 (positive control) group, rats were administered daily with lead acetate 50 mg/kg bw and distilled water four hours later; T1, T2, and T3 groups, rats were administered daily with lead acetate 50 mg/kg bw and 0.5 mL of 25, 50, and 100% white guava fruit juice four hours later. The treatment of the rats was conducted for 14 days, and on day 15, all rats were sacrificed to assess the spermatozoa quality. Data was analyzed using ANOVA followed by Duncan's multiple range test at a confidence level of 95%. The results showed that exposure to lead acetate (T0) caused lower spermatozoa PMI, viability, and motility as well as higher spermatozoa morphological abnormalities (p <0.05) compared to those of the T0 group. Administration of white guava fruit juice starting at a dose of 25% (T1) resulted in higher spermatozoa motility, viability, and PMI as well as lower spermatozoa morphological abnormalities (p <0.05) compared to rats in the T0 group. It could be concluded that white guava fruit juice maintained the spermatozoa quality of lead acetate induced rats.

Homoeopathic Management of Male Infertility—A Case Series

Abstract Background and Objectives Male infertility means a man is not able to start a pregnancy with his female partner. Infertility affects 10 to 15% of the couples globally. Male factor infertility contributes to about 20 to 30% of the cases. Low sperm concentration, poor sperm motility and abnormal sperm morphology are the main etiological factors for male infertility. For some patients, high cost of conventional treatment can be a deterrent while some fail to respond to it. Therefore, there is a need to explore the role of homoeopathy as an alternative. Methods Five male patients with infertility were treated with homoeopathic medicine based on the totality of symptoms after consulting repertory and Materia Medica. Further studies with suitable sample size are the cases were followed up with laboratory investigations are reported according to the HOM-CASE guidelines. Results Improvement was guaged by sperm analysis of all five men during treatment and all five cases resulted in pregnancy. Conclusion Further studies with suitable sample size are Warranted for further validation of the results.

Aberrant protamination in sperm correlates to anomalous nuclear and cytoplasmic architectures in infertile males with sperm dysmorphology.

Aberrant sperm protamination is linked to sperm dysmorphology and nuclear chromatin condensation. Yet, its effects on sperm cytoplasmic maturation remain largely unexplored. The relationships of protamines, sperm morphology, DNA damage, and cytoplasmic remodeling were illustrated in this study to provide fresh perspectives on the mechanisms of male infertility. A total of 205 infertile males were allocated into 5 groups according to the percentage of spermatozoa exhibiting abnormal morphology within their samples. Sperm concentration, motility, abnormal sperm morphology, cytoplasmic droplets (CDs), and excess residual cytoplasm (ERC) were analyzed according to the World Health Organization manual (2010). Sperm nuclear vacuoles (NVs) were determined by propidium iodide (PI) staining. Sperm protamine expressions (P1 and P2) were detected by western blot. DNA damage was measured by acridine orange test (AOT) to calculate the proportion of sperm with single-strand DNA breaks (SSBs). Our data showed that sperm concentration and motility in infertile males significantly decreased with the severity of abnormal sperm morphology (both P < 0.01). P1 level, P1/P2 ratio, and SSB rate increased with the severity of sperm dysmorphology, whilst the P2 level decreased (all P < 0.01). NVs, CDs, and ERC were more common in males with sperm dysmorphology and positively correlated with the SSB rate (all P < 0.01). The relationships between the SSB rate and the P1/P2 ratio were also significant ( P < 0.01). Aberrant protamination may cause sperm dysmorphology and compromise male fertility by impairing sperm's nucleus and cytoplasm maturation, with the P1/P2 ratio potentially serving as a valuable indicator of sperm quality and male fertility.

Rambutan Fruit Peel Extract Reduces Abnormal Sperm Morphology in Male Wistar Rats with Obesity

AbstractObesity is an accumulation of excessive fat tissue in the body. Excessive fat tissue in the body lead to infertility by increased Reactive Oxygen Species (ROS) and decrease the hormone balance regulation, those things can be affected the process of spermatogenesis, especially sperm morphology. Rambutan peel was known as a source of antioxidants because it has phenolic compounds, so it has a protective effect on free radicals. The research purpose knew the effect of Rambutan Peel Extract (RPE) (Nephelium lappaceum) on abnormal sperm morphology of Wistar rats (Rattus novergicus) percentage induced with a High-Fat Diet (HFD). This study uses True experimental post control group design for this research on 30 male Wistar rats. Samples were divided into 5 groups. Group 1: induced HFD only, Group 2: only given RPE at a dosage of 30 mg/kilogram Body Weight (kg BW), Group 3, 4, and 5: induced HFD and RPE at dosage of 15 mg/kg BW, 30 mg/kg BW, and 60 mg/kg BW feed using sonde. The data were analysed using ANOVA One Way. The result showed that RPE has decreased the abnormal sperm morphology of male Wistar rats at dose of 15 mg/kg BW. This is the first study that observe the effect of RPE administration to sperm morphology of obese and non-obese group of rats, with larger rats’ population, several doses of the RPE extract, and longer time to complete one cycle of rat spermatogenesis.AbstrakObesitas diartikan sebagai akumulasi jaringan lemak berlebihan yang ada di dalam tubuh. Jaringan lemak yang berlebihan dapat menyebabkan ketidakseimbangan regulasi hormonal dan terbentuknya Reactive Oxygen Species (ROS). Kedua hal tersebut dapat mempengaruhi proses spermatogenesis sehingga dapat menyebabkan infertilitas, terutama pada morfologi sperma. Ekstrak Kulit Rambutan (EKR) diketahui memiliki efek sebagai antioksidan dikarenakan memiliki senyawa fenolik, senyawa tersebut dapat menangkal radikal bebas. Penelitian ini dilakukan untuk mengetahui pengaruh pemberian EKR (Nephelium lappaceum) terhadap persentase morfologi abnormal sperma tikus Wistar (Rattus novergicus) yang diinduksi dengan Pakan Tinggi Lemak (PTL). Penelitian ini menggunakan desain True experimental post control group design pada 30 ekor tikus. Sampel dibagi menjadi 5 kelompok. Kelompok 1: hanya diberi PTL, Kelompok 2: hanya diberikan EKR dengan dosis 30 mg/kilogram Berat Badan (kgBB), Kelompok 3, 4 dan 5: diinduksi dengan PTL dan EKR dengan dosis 15 mg/kgBB, 30 mg/kgBB, dan 60 mg/kgBB. Pemberian PTL dan EKR dilakukan menggunakan sonde. Data dianalisis menggunakan uji ANOVA One Way. Hasil penelitian menunjukkan bahwa EKR dapat menurunkan jumlah morfologi abnormal pada kelompok dengan induksi PLT, dengan dosis yang paling efektif 15 mg/kgBB. Studi ini adalah yang pertama dalam mencari tahu efek pemberian RPE tehadap morfologi spermatozoa pada kelompok tikus obesitas dan tidak obesitas, dengan populasi tikus yang lebih besar, beberapa dosis ekstrak RPE, dan waktu yang lebih lama agar dapat menyelesaikan satu siklus spermatogenesis tikus.

A REVIEW OF PARACETAMOL: MECHANISM OF ACTION AND THE EFFECTS ON SPERMATOGENESIS

Nowadays, there are many factors causing the decrease of male reproductive function such as diet, routine, chemical, ... But some of them have not been paid appropriate attention to, especially the use of paracetamol, which has a significant negative effect on the reproductive process. In this study, we will summarize the results of several studies on the harmful effects of prolonged high-dose use of paracetamol on sperm number, quality, portability and testicular morphology in experimental animals and humans. The association between the long-term usage of paracetamol in chronic human diseases and their toxicity is highly concerned, especially the reproductive function. Their toxicity has increased more in adult male subjects, along with the introduction of studies on the use of high and long-term doses of paracetamol affecting reproductive quality. This review discusses the link between long-term usage of high doses of paracetamol and fertility as well as the effects of sex hormones in experimental animals and humans nowadays. As a result, paracetamol affects the reproductive system, impairing spermatogenesis and sperm quality. The amount of reproductive function damage, the dosage and duration of paracetamol usage are closely correlated. When groups of participants take paracetamol, numerous investigations in both experimental animals and humans have shown a decrease in sperm count and sperm motility as well as abnormalities in sperm morphology and testicular histology. Physicians have to exercise caution while treating patients with paracetamol and other related medications, especially men. Therefore, more studies should be done to determine the relationship between dosage, response, and treatment duration. When treating patients with decreased sexual function, medications that can prevent paracetamol toxicity should be used in conjunction with other medications. Especially, traditional medicine continues to play a significant part in the research and development of many new medicines despite the rapid advancement of contemporary medicine.

Nickel-induced oxidative stress causes cell death in testicles: implications for male infertility.

Aligarh region is well known for its lock industry. This lock industry utilises nickel for electroplating. There have been informal reports of infertility in men and women living near the lock industry. We analysed field water samples to investigate this link, and the results showed considerable nickel contamination. To further validate our results, we exposed male rats to relevant nickel levels in drinking water. This experimental exposure resulted in abnormal sperm morphology, decline in sperm count, significant change in activities of antioxidant enzymes, pronounced oxidative stress in the rat spermatocytes and decrease in serum testosterone level, as well as damage in the hypothalamus and pituitary (in all cases, the changes were most significant at the highest concentration used i.e 2.5 mg/l). The breeding experiments showed decline in live birth rate, while pups did not survive post birth in cages where males were given 2 and 2.5 mg/l concentrations of nickel in drinking water prior to mating. Our data strongly indicate a link between industrial nickel exposure and male infertility.

Effects of adverse semen parameters on total oxidation status and DNA/chromatin integrity

BackgroundInfertility is a major health problem that affects 7% of the men’s population. Oxidative stress (OS) plays a significant role in the pathophysiology of male infertility. The purpose of this study comparatively evaluated the total anti-oxidation status and DNA/chromatin integrity in semen samples among different infertile men’s groups compared with the normozoospermic men.MethodsThis cross-sectional study contains four experimental groups, including teratozoospermia (Exp I), asthenoteratozoospermia (Exp II), oligoasthenoteratozoospermia (Exp III), and azoospermia (Exp IV), as well as the control group of normozoospermic men. The total antioxidant capacity (TAC) and total oxidant status (TOS) were assessed by applying the enzyme-linked immunosorbent assay. The chromatin/DNA damage was assessed in semen samples of all study groups by applying chromomycin A3 (CMA3) and toluidine blue (TB) staining methods.ResultsThe results showed significantly higher proportions of TB+ and CMA3 positive sperm in all experimental groups compared to controls (P < 0.001). TAC, TOS, and the ratio of TAC to TOS were significantly different in all experimental groups compared to the normozoospermic men (P < 0.001).ConclusionOur study demonstrated that at least one sperm parameter abnormality, such as teratozoospermia could cause serious defects at the levels of DNA/chromatin as well as the antioxidants to oxidant balance of human spermatozoa in subfertile men with abnormal spermogram. Infertile men with sperm morphological abnormalities may benefit from simultaneous assessment of sperm DNA defects and OS.

APOLLO, a testis-specific Drosophila ortholog of importin-4, mediates the loading of protamine-like protein Mst77F into sperm chromatin

DNA in sperm is packed with small, charged proteins termed SNBPs (sperm nuclear basic proteins), including mammalian and Drosophila protamines. During spermiogenesis, somatic-type chromatin is taken apart and replaced with sperm chromatin in a multistep process leading to an extraordinary condensation of the genome. During fertilization, the ova face a similarly challenging task of SNBP eviction and reassembly of nucleosome-based chromatin. Despite its importance for the animal life cycle, sperm chromatin metabolism, including the biochemical machinery mediating the mutual replacement of histones and SNBPs, remains poorly studied. In Drosophila, Mst77F is one of the first SNBPs loaded into the spermatid nuclei. It persists in mature spermatozoa and is essential for sperm compaction and male fertility. Here, by using invitro biochemical assays, we identify chaperones that can mediate the eviction and loading of Mst77F on DNA, thus facilitating the interconversions of chromatin forms in the male gamete. Unlike NAP1 and TAP/p32 chaperones that disassemble Mst77F-DNA complexes, ARTEMIS and APOLLO, orthologs of mammalian importin-4 (IPO4), mediate the deposition of Mst77F on DNA or oligonucleosome templates, accompanied by the dissociation of histone-DNA complexes. Invivo, a mutation of testis-specific Apollo brings about a defect of Mst77F loading, abnormal sperm morphology, and male infertility. We identify IPO4 ortholog APOLLO as a critical component of sperm chromatin assembly apparatus in Drosophila. We discover that in addition to recognized roles in protein traffic, a nuclear transport receptor (IPO4) can function directly in chromatin remodeling as a dual, histone- and SNBP-specific, chaperone.

A variant in sperm-specific glycolytic enzyme enolase 4 (ENO4) causes human male infertility.

Although defects in sperm morphology and physiology lead to male infertility, in many instances, the exact disruption of molecular pathways in a given patient is often unknown. The glycolytic pathway is an essential process to supply energy in sperm cell motility. Enolase 4 (ENO4) is crucial for the glycolytic process, which provides the energy for sperm cells in motility. ENO4 is located in the sperm principal piece and is essential for the motility and organization of the sperm flagellum. In the present study, we characterized a family with asthenozoospermia and abnormal sperm morphology as a result of a variant in the enolase 4 (ENO4) gene. Computer-assisted semen analysis, papanicolaou smear staining and scanning electron microscopy were used to examine sperm motility and morphology for semen analysis in patients. For genetic analysis, whole-exome sequencing followed by Sanger sequencing was performed. Two brothers in a consanguineous family were being clinically investigated for sperm motility and morphology issues. Genetic analysis by whole-exome sequencing revealed a homozygous variant [c.293A>G, p.(Lys98Arg)] in the ENO4 gene that segregated with infertility in the family, shared by affected but not controls. In view of the association of asthenozoospermia and abnormal sperm morphology in Eno4 knockout mice, we consider this to be the first report describing the involvement of ENO4 gene in human male infertility. We also explore the possible involvement of another variant in explaining other phenotypic features in this family.

Morphological analyses of sperms causing severe male infertility with light microscopy, motile sperm organellar morphology examination and electron microscopy tecnics.

To evaluate abnormal sperm morphology by three different methods including light microscope, real-time motile sperm organelle morphology assessment at 8050x high magnification and electron microscope and to determine degree of affect on morphologic defects level. Beside the light and electron microscopic techniques, the motile sperm organelle examination can offer information about sperm quality in a simple and quick way and by the IMSI developed through the use of sperm selected by this technique can contribute to the success of in vitro fertilization centers. We believe that sperm selection at high magnifications can increase IVF pregnancy rates in patients with sperm morphology that causes severe male infertility.

Protective effects of melatonin against oxidative stress induced by metabolic disorders in the male reproductive system: a systematic review and meta-analysis of rodent models.

Male infertility is a multifaceted issue that has gained scientific interest due to its increasing rate. Studies have demonstrated that oxidative stress is involved in male infertility development. Furthermore, metabolic disorders, including obesity, diabetes, hypo- and hyperthyroidism, are risk factors for male infertility, and oxidative stress is believed to contribute to this association. Melatonin, functioning as an oxidative scavenger, may represent a promising therapeutic approach for the prevention and treatment of metabolic disorder-associated male infertility. We systematically searched three online databases (PubMed, Scopus, and Web of Science) for studies that evaluated the effects of melatonin therapy on metabolic disorders-induce infertility in male rodents. The favorable outcomes were histopathological parameters of testicular tissue, reproductive hormones, and markers of oxidative stress. Then, meta-analyses were done for each outcome. The results are reported as standardized mean difference (Cohen's d) and 95% confidence interval. 24 studies with 31 outcomes were included. Rats and mice were the subjects. Studies have employed obesity, diabetes, hypothyroidism, hyperthyroidism, hyperlipidemia, and food deprivation as metabolic disorders. To induce these disorders, a high-fat diet, high-fructose diet, leptin, streptozotocin, alloxan, carbimazole, and levothyroxine were used. The outcomes included histopathologic characteristics (abnormal sperm morphology, apoptotic cells, apoptotic index, Johnsen's testicular biopsy score, seminiferous epithelial height, tubular basement membrane thickness, tubular diameter, sperm count, and motility), weight-related measurements (absolute epididymis, testis, and body weight, body weight gain, epididymal adipose tissue weight, and relative testis to body weight), hormonal characteristics (androgen receptor expression, serum FSH, LH, and testosterone level), markers of oxidative stress (tissue and serum GPx and MDA activity, tissue CAT, GSH, and SOD activity), and exploratory outcomes (serum HDL, LDL, total cholesterol, triglyceride, and blood glucose level). The overall pooled effect sizes were statistically significant for all histopathological characteristics and some markers of oxidative stress. Melatonin can reduce damage to male rodents' gonadal tissue and improve sperm count, motility, and morphology in metabolic diseases. Future clinical studies and randomized controlled trials are needed to evaluate the safety and effectiveness of melatonin for male infertility in patients with metabolic diseases.

Therapeutic effects of melatonin in long-term exposure to 2100 MHz radiofrequency radiation on rat sperm characteristics

IntroductionRadiofrequency electromagnetic fields (RF-EMFs) are one of the risk factors for male reproductive health and melatonin can be an ideal candidate for therapeutic development against RF-induced male fertility problems due to its antioxidant properties. The possible therapeutic role of melatonin in the destructive effects of 2100MHz RF radiation on rat sperm characteristics is investigated in the present study. MethodsWistar albino rats were divided into four groups and the experiment continued for ninety consecutive days; Control, Melatonin (10mg/kg, subcutaneously), RF (2100MHz, thirty minutes per day, whole-body), and RF+Melatonin groups. Left caudal epididymis and ductus deferens tissues were placed in sperm wash solution (at 37°C) and dissected. The sperms were counted and stained. Measurements of the perinuclear ring of the manchette and posterior portion of the nucleus (ARC) were performed and the sperms were examined at an ultrastructural level. All of the parameters were evaluated statistically. ResultsThe percentages of abnormal sperm morphology were significantly increased with RF exposure, while the total sperm count was significantly decreased. RF exposure also showed harmful effects on acrosome, axoneme, mitochondrial sheath, and outer dense fibers at the ultrastructural level. The number of total sperms, sperms with normal morphology increased, and ultrastructural appearance returned to normal by melatonin administration. DiscussionThe data showed that melatonin may be a beneficial therapeutic agent for long-term exposure of 2100MHz RF radiation-related reproductive impairments.

P-024 Effect of microsurgical varicocelectomy on fertility potential in patients with low sperm vitality

Abstract Study question Does microsurgical varicocelectomy improve semen parameters and fertility potential in patients with low sperm vitality? Summary answer Micro surgical varicocelectomy in patients with low sperm vitality is beneficial as it results in significant improvement in sperm count and progressive motility. What is known already Low sperm vitality (Necrozoospermia) indicates the presence of higher percentage of necrotic/ dead sperm in the ejaculate which can impair fertility potential. Varicocele induced testicular hyperthermia is among the most common causes of impaired semen quality, including necrozoosprmia. While many studies confirmed the beneficial effect of varicocelectomy on different semen parameters, very few have evaluated its role on sperm vitality in particular. Study design, size, duration A retrospective study of 1521 patients who underwent microsurgical sub inguinal varicocelectomy in our center from 2011–2021. A total of 115 patients with clinical varicocele and low sperm vitality (&amp;lt;58%) were included. Participants/materials, setting, methods The exclusion criteria were history of genitourinary tract infection, exposure to heat, drugs or toxic compounds, hormonal disturbances or prior infertility related treatments. Patients demographics, and clinical data were collected, in addition to the pre- and post-operative Semen analysis (WHO 5th edition), sperm DNA fragmentation (Halosperm) and hormonal profile (estradiol, FSH, LH, Prolactin, Testosterone). Spearman’s correlations and Wilcoxon Signed Ranked Test were used for data analysis. Main results and the role of chance Mean age was 34.9±7.6 years. Varicocelectomy was done on left side in 75 patients and bilateral in 40 patients. Sperm vitality was significantly negatively correlated with sperm DNA fragmentation (-0.618, p = 0.000) and abnormal sperm morphology (-0.435, p = 0.000, while it was significantly positively correlated with total motility (0.674, p = 0.000) &amp; progressive motility (0.272, p = 0.003). The median (IQR) sperm vitality pre-operatively was 41% (30-50) and the level increased post-operatively to 54% (31-63). There was a significant improvement in post-operative sperm count, progressive motility as well as LH and FSH levels. Improvements were also noted in total motility, normal morphology and sperm DNA fragmentation however the result didn’t reach statistical significance. Limitations, reasons for caution The main limitation is the retrospective design of the study and it did not evaluate the pregnancy outcomes. Wider implications of the findings Improvement in semen parameters by microsurgical varicocelectomy in patients with low sperm vitality is important as it may change the management plan for patients and extend their fertility options to natural conception or intrauterine insemination instead of only intracytoplasmic sperm injection. Trial registration number not applicable

P-069 A male factor panel of NGS can identify the genetic cause of more than half patients with seminal alterations

Abstract Study question Is Next Generation Sequencing (NGS) an effective diagnostic tool for sperm abnormalities? Summary answer NGS male factor gene panel can be a potent diagnostic tool for discovering pathogenic variants linked to sperm abnormalities. What is known already Male infertility, responsible for half of infertility cases, often shows as absence or decreased sperm count (azoospermia, criptozoospermia or oligozoospermia), poor motility (asthenozoospermia) or a higher proportion of morphologically abnormal sperm (teratozoospermia). While the spermiogram remains the gold standard for evaluating male fertility, the potential to identify the molecular basis of sperm abnormalities may make NGS a valuable approach for such patients. The aim of our study was to assess NGS panel's effectiveness in diagnosing patients with abnormal sperm count, motility and morphology. Study design, size, duration A prospective study was conducted from May 2021 to May 2022 in 82 patients including 20 patients with oligozoospermia, 19 with asthenozoospermia and 23 with teratozoospermia. No other sperm alterations were taken into account in each group. The control group consisted of 20 normozoospermic healthy donors selected on the basis of normal sperm parameters according to the WHO criteria (2010). Patients carrying Y-chromosome microdeletions or abnormal karyotype were excluded. Participants/materials, setting, methods Genomic DNA extraction from blood-EDTA of the patients was performed using the commercial MagMax DNA MultiSample Ultra kit and the King-Fisher automated extractor (ThermoFisher®). Next Generation Sequencing (NGS) was done using a panel with 426 genes involved in the spermatogenesis process. Panel sequencing for identification of genetic variants was performed using Nextera Enrichment technology (Illumina®). FASTAQ data were processed using BWA and GATK algorithms. VCF files were analysed using Variant Interpreter software and in silico predictors. Main results and the role of chance Data analysis showed that thirty-seven of the sixty-two patients were carriers of pathogenic mutations in at least one of the genes included in the panel (37/62,59.6%). In the oligozoospermic group, eleven patients (11/20,55%) were carriers of pathogenic mutations in:CFTR, CEP290, WDR66, ESR1, DNAI2, POLG, PIWIL3, GNRHR, MSH5 and CTNS. In the asthenozoospermic group, ten out of nineteen patients (10/19,52.6%) were carriers of pathogenic mutations in:HS6ST1, PMS2, CYP19A1, DNAI2, POLG, LRRC6, G6PD, CCDC39 and PIWIL3. In the teratozoospermic group, six of the twenty-three patients (6/23,26%) were carriers of pathogenic mutations in:CFTR, PIWIL3, CYP21A2, SRD5A. No pathogenic mutations were detected in the control group. The analysis showed comparable results among the various spermiogram alterations with no significant difference (p &amp;gt; 0.05). Each variant was found in a single patient per group, except POLG and CCDC39, which were found in two cases in the oligozoospermic and asthenozoospermic group, respectively, also CFTR and PIWIL3 in two teratozoospermic patients. Besides, it is remarkable that some of the pathogenic variants were present in genes in more than one group:CFTR (3/62), DNAI2 (2/62), POLG (3/62) and PIWIL3 (4/62). All identified variants were linked with processes of spermatogenesis such as dynein assembly and DNA integrity. Limitations, reasons for caution The main limitation of this study is the limited number of patients included. Functional studies with a larger cohort of males with seminal disorders is warranted to confidently correlate the genetic variants identified in this analysis with spermatogenic failure. Wider implications of the findings The gene list included in our panel represents a step-forward in the screening of males with altered sperm parameters. Our results may add in the knowledge of male factor infertility to provide etiologic factors towards a personalized treatment and adequate genetic counselling. Trial registration number Not applicable