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Necrozoospermia is a poorly documented condition with a low incidence, and its definition and clinical significance are unclear. Herein, we provide a reference range for necrozoospermia and discuss its possible etiology and impact on male fertility and assisted reproductive outcomes. We extracted relevant information from 650 Chinese male partners of infertile couples and statistically analyzed sperm vitality. Necrozoospermia was present in 3.4% (22/650) of our study population, and the lower cut-off value for sperm vitality was 75.3%. We compared two methods for assessing sperm vitality (eosin-nigrosin head staining and hypo-osmotic swelling test [HOST]), for which the percentage in the eosin-nigrosin group (mean ± standard deviation [s.d.]: 77.5% ± 10.5%) was significantly higher than that in the HOST group (mean ± s.d.: 58.1% ± 6.7% [5–10 min after incubation] and 55.6% ± 8.2% [25–30 min after incubation]; both P < 0.001). The incidence of necrozoospermia increased with age (odds ratio [OR] = 1.116, 95% confidence interval [CI]: 1.048–1.189, P = 0.001), while the percentage of normal sperm morphology and DNA fragmentation index (DFI) were significantly associated with necrozoospermia, with ORs of 0.691 (95% CI: 0.511–0.935, P = 0.017) and 1.281 (95% CI: 1.180–1.390, P < 0.001), respectively. In the following 6 months, we recruited 166 patients in the nonnecrozoospermia group and 87 patients in the necrozoospermia group to compare intracytoplasmic sperm injection (ICSI) and pregnancy outcomes between the two groups. The necrozoospermia group had a significantly lower normal fertilization rate (74.7% vs 78.2%, P = 0.041; OR = 0.822; 95% CI: 0.682–0.992) than that in the nonnecrozoospermia group. This study presents substantial information on necrozoospermia to establish comprehensive and applicable reference values for sperm vitality for spontaneous conception and artificially assisted reproductive management.

Background: Despite the clear clinical diagnostic criteria for necrozoospermia in andrology, the fundamental mechanisms underlying it remain elusive. This study aims to profile the lipid composition in seminal plasma systematically and to ascertain the potential of lipid biomarkers in the accurate diagnosis of necrozoospermia. It also evaluates the efficacy of a lipidomics-based random forest algorithm model in identifying necrozoospermia. Methods: Seminal plasma samples were collected from patients diagnosed with necrozoospermia (n = 28) and normozoospermia (n = 28). Liquid chromatography–mass spectrometry (LC–MS) was used to perform lipidomic analysis and identify the underlying biomarkers. A lipid functional enrichment analysis was conducted using the LION lipid ontology database. The top 100 differentially significant lipids were subjected to lipid biomarker examination through random forest machine learning model. Results: Lipidomic analysis identified 46 lipid classes comprising 1267 lipid metabolites in seminal plasma. The top five enriched lipid functions as follows: fatty acid (FA) with ≤ 18 carbons, FA with 16–18 carbons, monounsaturated FA, FA with 18 carbons, and FA with 16 carbons. The top 100 differentially significant lipids were subjected to machine learning analysis and identified 20 feature lipids. The random forest model identified lipids with an area under the curve > 0.8, including LPE(20:4) and TG(4:0_14:1_16:0). Conclusions: LPE(20:4) and TG(4:0_14:1_16:0), were identified as differential lipids for necrozoospermia. Seminal plasma lipidomic analysis could provide valuable biochemical information for the diagnosis of necrozoospermia, and its combination with conventional sperm analysis may improve the accuracy and reliability of the diagnosis. [ABSTRACT FROM AUTHOR]

A major factor affecting male fertility is excessive death of germ cells, both immature germ cells and mature spermatozoa. It can be due to various factors causing testicular and/or post-testicular damage, such as infections, obstructive conditions, toxins, oxidative stress, hormonal imbalance, hyperthermia, and anti-sperm antibodies. Massive death of spermatozoa leads to a high proportion of dead sperm cells in the ejaculate (necrozoospermia or necrospermia) while death of immature germ cells can lead to low sperm count (oligozoospermia or oligospermia). Cell death can occur both by necrosis and by apoptosis; in recent decades, it has been found that apoptosis of mature spermatozoa is not only possible but quite common, and can contribute to infertility. Treatment approaches are primarily directed to the underlying condition, i.e. removing the cause(s) of sperm cell death whenever possible, but include also attempts to bypass the cell death event by intracytoplasmic sperm injection with testicular spermatozoa.

Summary: Spermiostasis (or sperm accumulation) is an ejaculatory dysfunction of common occurrence in referral reproductive practice with stallions. The accumulation of sperm in the epididymides, ducti deferens, and ampullae results in ejaculates with a high concentration of degenerated sperm. It is estimated that 5% of light‐breed stallions present this condition; however, the incidence is unknown in ponies as this condition has not been reported in these breeds. The present report describes the diagnosis and management of spermiostasis in a 16‐year‐old Shetland stallion presented for semen collection, evaluation, and cooling test. Semen collection resulted in 73 billion total sperm with necrozoospermia and 67% detached heads. Ten serial semen collections gradually improved sperm motility and morphology. There was no evidence of ampullary plugs in the semen filter throughout the collections. An additional semen collection was procured 48 h after collection 10 for a cooling test. Sodium caseinate cholesterol‐loaded cyclodextrin extender resulted in superior sperm motility compared to the native phosphocaseinate extender up to 48 h post‐cooling in two semen containers. Sixty days after the initial visit, the stallion returned for a semen freezing test. The ejaculate was normozoospermic, and the owner reported that the stallion had satisfactory fertility for live cover after discharge from the hospital. The thermal longevity test of the frozen‐thawed resulted in excellent motility after 120 min of storage at 37°C. In conclusion, herein, we described, apparently for the first time, the occurrence of spermiostasis in a pony stallion. We foresee that other practitioners managing similar clinical cases could have comparable outcomes. [ABSTRACT FROM AUTHOR]

According to some statistics, absolute asthenozoospermia affects every 1 in 5000 men. Although this incidence rate does not appear to be too high, it is extremely important to address the phenomenon because it can drastically reduce the chances of pregnancy, even with assisted reproduction. The biggest problem with absolute asthenozoospermia is that it is difficult to distinguish between live and dead sperm cells, and fertilization with non-viable spermatozoa may contribute to the failure of an assisted reproduction cycle. Nowadays, DNA fragmentation (DF) is a crucial parameter of semen analysis, and in this paper, we provide evidence of the correlation between DF and vitality. For this purpose, the main semen parameters were investigated by a CASA system (concentration, motility, progressive motility, vitality and DF). In the necrozoospermic group (vitality < 58%), all the measured parameters showed significant differences compared to normal vitality. Concentration (30.1 M mL−1 vs. 13.6 M mL−1), motility (31.9% vs. 18.3%), and progressive motility (24.3% vs. 12.7%) were significantly decreased, while DF was significantly increased (17.4% vs. 23.7%). Based on the connection between vitality decrement and DF increment, DF lowering methods, such as magnetic-activated cell sorting, have been hypothesized as novel methods for the elimination of dead spermatozoa. [ABSTRACT FROM AUTHOR]

Background: Necrozoospermia is a condition found in 0.2%–0.4% of male infertility cases. The causes of necrozoospermia are multiple: they can be related to testicular and/or post‐testicular damage. Additionally, these causes most often involve the production of reactive oxygen species (ROS) and/or sperm DNA fragmentation (SDF) which can reduce the chances of spontaneous pregnancy or affect the outcome of assisted reproductive technologies. Objective: To focus on potential etiologies of necrozoospermia, its diagnosis and its therapeutic management especially before the employment of ICSI. Methods: Authors searched PubMed/Medline, Web of Science, Cochrane Library, Google and Institutional websites for medical subheading terms and free text words referred to "necrozoospermia", "sperm vitality", "sperm viability", SDF and "ICSI". Results: We identified 12 main etiologies of necrozoospermia responsible for either a decrease of sperm vitality, a mild, a moderate or a severe necrozoospermia. In case of a confirmed decreased vitality, a thorough check‐up should be conducted and if available, etiological treatment should be proposed. Therapeutic management could also include repeated ejaculations, drug treatments, the use of ICSI with ejaculated or surgically extracted spermatozoa in case of a non‐treatable necrozoospermia. Discussion and conclusion: The potential causes of necrozoospermia should be investigated because many of them could be corrected, thus avoiding the use of ICSI. Moreover, if ICSI procedure remains necessary, the therapeutic management of necrozoospermia could also improve the chances of success by reducing oxidative stress and/or SDF. [ABSTRACT FROM AUTHOR]

Asthenozoospermia is the most frequent sperm motility disorder, but there are other more extreme sperm motility disorders, namely oligo-astheno-teratozoospermia (OAT) and necrozoospermia. There are several cellular mechanisms known for OAT and necrozoospermia, but there are limited data on dynein ATPase and ATPases associated with various cellular activities (AAA+). AAA1 is involved in ATP hydrolysis, while AAA2 is entangled in ATP-binding pocket. This study was conducted to investigate the role of dynein ATPase activity and quantification of AAA dynein. Spermatozoa from 14 men with OAT, 11 men with necrozoospermic and 17 men with normoozspermic samples were used in this study. Makler chamber was used to determine sperm concentration and motility, while Papanicolaou stained semen smears using World Health Organization-fifth edition criteria was performed to determine sperm morphology, and dynein ATPase was quantified by calculation of released inorganic phosphate. AAA was quantified by enzyme-linked immunosorbent assay, whereas the distribution was determined by immunocytochemistry. This study showed that the dynein ATPase activity in OAT and necrozoospermia was lower than in the normozoospermic group (2.68±0.76, 1.01±0.31, 7.22±1.08 μmol Pi/mg protein/h, respectively, P

Asthenozoospermia is the most frequent sperm motility disorder, but there are other more extreme sperm motility disorders, namely oligo-astheno-teratozoospermia (OAT) and necrozoospermia. There are several cellular mechanisms known for OAT and necrozoospermia, but there are limited data on dynein ATPase and ATPases associated with various cellular activities (AAA+). AAA1 is involved in ATP hydrolysis, while AAA2 is entangled in ATP-binding pocket. This study was conducted to investigate the role of dynein ATPase activity and quantification of AAA dynein. Spermatozoa from 14 men with OAT, 11 men with necrozoospermic and 17 men with normoozspermic samples were used in this study. Makler chamber was used to determine sperm concentration and motility, while Papanicolaou stained semen smears using World Health Organization-fifth edition criteria was performed to determine sperm morphology, and dynein ATPase was quantified by calculation of released inorganic phosphate. AAA was quantified by enzyme-linked immunosorbent assay, whereas the distribution was determined by immunocytochemistry. This study showed that the dynein ATPase activity in OAT and necrozoospermia was lower than in the normozoospermic group (2.68±0.76, 1.01±0.31, 7.22±1.08 µmol Pi/mg protein/h, respectively, P<0.05), as well as the amounts of AAA1 and AAA2. In addition, staining for AAA in the sperm tail paralleled the dynein ATPase activity and quantity of AAA, being the highest in sperm from normozoospermic samples, lower in sperm from OAT samples, and almost undetectable in sperm from necrozoospermic samples. The structure and function of damaged sperm dynein may alter dynein ATPase activity and levels of AAA1 and AAA2. [ABSTRACT FROM AUTHOR]

Male factors are responsible for more than forty percent of the infertility cases. Necrozoospermia is one among the main cause for infertility in male. Necrospermia i.e. 100% immotile and 0% viable spermatozoa in ejaculate, is a rare and poorly documented cause of male infertility. A 56-year-old man was referred for reproductive counseling with secondary infertility and diagnosed as necrozoospermia (Shukra Dosha). He presented with complaints of having no issues since 7 years of married life with his second wife. The patient has a son from divorced first wife. After the thorough clinical examination and laboratory investigations, diagnosis was confirmed as necrozoospermia. The patient was administered Koshtha-Shuddhi (purgation) with Eranda Taila (castor oil) as per the guidelines of Ayurvedic treatment. After the proper purificatory procedures depending on dominance of deranged Dosha, the patient was given Arogyavardhini Vati, Chandraprabha Vati, Shilapravanga and Phala Ghrita (medicated ghee) as Sneha (internal oleation). At the end of 3½ month, semen analysis showed marked improvement in sperm count and increase in the sperm motility. The present finding and the effective management of necrozoospermia with Ayurvedic formulations with no adverse effect highlight the promising scope of traditional medicine in male infertility disorders. [ABSTRACT FROM AUTHOR]

Background: In the pathological study of necrozoospermia-a form of sperm mortality-the underlying metabolic mechanism remains unclear. Thus, the aim of this study was to characterize metabolic alterations in the seminal plasma of necrozoospermic patients and to provide insights into the etiology of the disease., Methods: Seminal plasma samples were collected from patients diagnosed with necrozoospermia (n=28) as well as normozoospermia (n=37). The samples were analyzed using nontargeted metabolomics based on liquid chromatography-mass spectrometry (LC-MS). The raw data were subjected to multivariate analysis to identify metabolites correlated with necrozoospermia. Differential metabolites were subjected to pathway analysis using MetaboAnalyst., Results: The results of the metabolomic analysis showed that there were 194 differential metabolites between the two groups; 129 metabolites were upregulated and 65 metabolites were downregulated. Among the differential metabolites, the top ten differential metabolites were choline, benzaldehyde, pyrazinamide, 5-aminoimidazole-4-carboxamide, and dihydrothymine. The following differential metabolite pathways were identified, and the top five metabolite pathways were arachidonic acid metabolism, steroid hormone biosynthesis, alanine aspartate and glutamate metabolism, bile secretion, and prostate cancer., Conclusions: The elevation of choline and 2-hydroxyglutarate levels in seminal plasma was an important finding, and the results also indicate that abnormalities in arachidonic acid metabolism and glutamate metabolism were an underlying pathological mechanism of necrozoospermia., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at https://tau.amegroups.com/article/view/10.21037/tau-23-14/coif). The authors have no conflicts of interest to declare., (2023 Translational Andrology and Urology. All rights reserved.)

Failure or severe difficulty in conceiving a child is surprisingly common, worldwide problem. Half of these cases are due to male factors with defects in sperm (1 in 15 men) being the single most common cause. Also about 60-75 % of male infertility cases are idiopathic, since the molecular mechanisms underlying the defects remain unknown. DNA methylation is crucial for spermatogenesis and high methylenetetrahydrofolate reductase (MTHFR) activity in adult testis than other organs in mouse, signifies its critical role in spermatogenesis. According to recent findings there is a correlation of epigenetic regulation of several imprinted genes with disturbed spermatogenesis and fertility. Consequently any change in the MTHFR gene sequence can modify the spermatogenesis including transmission of infertility to the carriers. The aim of the study is to analyze the distribution of the single nucleotide polymorphism C677T in the MTHFR gene in 637 North Indian infertile patients and 364 fertile North Indian men as controls by using PCR-RFLP technique and Chi Square test for statistical analysis. The average MTHFR 677CC, 677CT, 677TT genotype frequencies of total infertile men were 70.17, 24.17, 5.65 % in infertile men and 75.27, 21.7, 2.74 % in controls, respectively. The average frequency of the MTHFR 677T allele was 17.73 % in infertile men as compared to 13.59 % in controls. The statistical difference was significant. Disease risk was found 2.27-folds increased in patients who were carrying T allele. We found an association of C677T polymorphism with male infertility and that it may be a genetic risk factor for male infertility in North Indian population. [ABSTRACT FROM AUTHOR]

Objective: To determine whether intracytoplasmic sperm injection (ICSI) outcome with testicular sperm is superior to that of ejaculated sperm in men with incomplete necrozoospermia, defined as sperm viability ≥5 to ≤45%. Study design: Retrospective study at a Reproductive Medicine Center of a tertiary referral hospital. A total of 231 couples with male infertility due to incomplete necrozoospermia underwent 342 ICSI cycles with freshly ejaculated sperm (ICSI-ejaculated) and 182 cycles with testicular sperm (ICSI-TESE). Results: Overall 1624 MII oocytes were injected in the ICSI-ejaculated group with a fertilisation rate of 60.8%, while in ICSI- TESE cycles the fertilisation rate was 59.6% in 874 MII oocytes. The cleavage rate was higher in ICSI-ejaculated cycles than in ICSI-TESE cycles (96.3% versus 92.9%; p = 0.004). However, the pregnancy and implantation rates per cycle were significantly higher in the ICSI-TESE group (67/182, 36.8% versus 68/342, 19.9% ( p = 0.0001); and 23.7% versus 12.7% ( p = 0.0001), respectively). The miscarriage rate was similar (ICSI-ejaculated 26.5% versus ICSI-TESE 17.9%, p = 0.301). Live birth rate per cycle in the ICSI-ejaculated group was significantly lower than in the ICSI-TESE (13.7% versus 28.6%, p = 0.0001). Conclusions: In cases of persistent necrozoospermia, testicular sperm should be favoured over ejaculated sperm. These data call for more research to understand the pathophysiology of refractory necrozoospermia. [ABSTRACT FROM AUTHOR]

Introduction: Sperm apoptosis is well characterized but studies on the effect of male age and necrozoospermia are lacking. The objectives were: (a) to analyze percentages of apoptotic and necrotic sperm in ejaculates, and (b) to compare the results between younger and older age groups. Materials and Methods: Routine semen analyses were carried out (n = 189 males) and sperm cells were analyzed by dual fluorescence assay Hoechst 33342 and propidium iodide, and the acridine orange test. Results: The percentage of necrotic sperm in the ejaculate increased by 22% for males aged over 35. There was a positive correlation between age and necrosis (R = 0.30). Sperm apoptosis increased by 17% in males aged 45 and older. The population of DNA intact sperm declined in males aged 40 and over (R = –0.21). There were no age-related changes in strict normal morphology, sperm concentration and semen volume. A decrease in rapid progressive motility was correlated (R = –0.24) with male age and was significant after age 35. Conclusions: The study demonstrated increased necrosis, DNA damage and apoptosis while rapid progression and total motility declined with advancing age in the male beginning as early as age 35. The order of the observed changes was sequential, suggesting the involvement of different pathways in sperm necrosis after age 40. Copyright © 2007 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

We present the ultrastructural, functional and chromosomal analyses of sperm from an infertile man with a normal karyotype and 100% necrozoospermia. Tests for microbial infection showed the presence of Escherichia coli in seminal and urethral fluid. Semen analysis was performed concomitantly with the infection and repeated twice after the therapy, in the absence of infection, to control the possible recovery of necrozoospermia. Morphological sperm evaluation was performed by light, fluorescent and electron microscopy; meiotic segregation was examined by fluorescence in situ hybridisation (FISH) technique. Polymerase chain reaction (PCR) was carried out on DNA from peripheral blood lymphocytes to analyse partial sequences of the Akap4 and Akap3 genes. AKAP3 protein is synthesised in round spermatids, incorporated into the fibrous sheath and is involved in organising the basic structure of the fibrous sheath. AKAP4 protein is incorporated late in spermatid development and plays a major role in completing fibrous sheath assembly. The whole sperm population was immotile and non-viable. The ultrastructural characteristics of sperm necrosis were identified. Immunostaining of tubulin and AKAP4 proteins was negative in sperm tails. PCR did not reveal any deletions in the investigated regions. FISH sperm analysis highlighted an altered meiotic segregation. After recovery from infection, 100% necrozoospermia persisted and chromosomal sperm aneuploidies were still present. [ABSTRACT FROM AUTHOR]

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