Ejaculate Quality Research Articles

Ejaculate Adjustment in Response to Sperm Competition Risk in Humans.

Previous research suggests that human males, like males of many mammalian and avian species, adjust their ejaculate quality in accordance with sperm competition risk. Men who spend less time with their regular female partner since the couple's last copulation produce ejaculates with more sperm at the couple's next copulation (Baker & Bellis, 1993). We conducted a conceptual replication of this research to investigate whether sperm competition risk predicts ejaculate adjustment in human males using additional measures of sperm competition risk (e.g., perceptions of partner infidelity, presence of potential sexual rivals) and updated laser-optic semen analysis technology. We collected data from 34 heterosexual couples (age range 18-32years) from a university population who completed self-report surveys on their relationship dynamics and provided six ejaculate samples (three copulatory and three masturbatory) across a 45-day period. Time spent together since the couple's last copulation was not significantly associated with ejaculate quality. However, sperm concentration for copulatory ejaculates was higher for men who perceived more potential sexual rivals. Discussion situates the current results within the literature on human sperm competition and suggests several directions for future research.

How do taurine and ergothioneine additives improve the parameters of high- and low-quality turkey semen during liquid preservation?

Abstract The study aimed to examine the parameters of turkey ejaculates (n=40) of high and low quality (HQ and LQ, respectively), preserved with the addition of taurine (TAU; 5 mM, 10 mM) and ergothioneine (EGT; 5 mM, 10 mM) for 48 h at 5°C in a liquid state. The motility, plasma membrane integrity (PMI), mitochondrial function, apoptotic and necrotic spermatozoa percentage, and sperm cells that generated NO were determined after 2, 24, and 48 h of storage. The preserved semen was also analysed for the activity of superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), glutathione (GSH) content, and malondialdehyde (MDA) production. Taurine, in both doses, may improve the antioxidant status of stored turkey semen as well as sperm motility, viability and functionality regardless of ejaculate quality, as manifested by increased SOD and CAT activities, reduced MDA levels, and enhanced sperm characteristics i.e. plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), total motility (TMOT), and progressive motility (PMOT). In turn, the addition of EGT increased GSH content in the external environment and suppressed lipid peroxidation in turkey spermatozoa, in particular those from low-quality (LQ) ejaculates. However, it appears that EGT could deliver more satisfactory results when added at lower concentrations.

Expression of exosomal microRNAs miR-34a and miR-210 in male infertility: relationship with morphokinetic parameters and sperm DNA fragmentation

Introduction. Infertility affects tens of millions of men and women across the globe. In approximately half of cases, male factors are the cause of infertility. In recent decades, there has been a significant decline in the quality of male ejaculate, which is characterized by reduced sperm concentration and motility. The insufficient diagnostic and prognostic value of routine semen analysis results highlights the challenge of developing effective diagnostic tools and searching for reliable biomarkers of male infertility. One of the most promising approaches may be assessing sperm microRNA expression.Objective. To study the role of sperm exosomal microRNAs miR-34a and miR-210 in the development of male infertility.Materials & methods. The retrospective study included 150 men aged 25 – 49 years; of these, 96 patients were diagnosed with idiopathic infertility. The comparison group consisted of 54 fertile men. To assess the structure and motility of sperm, the results of a standard ejaculate study (WHO, 2021) and computer data analysis using MMC Sperm (MMCSoft, St. Petersburg, Russia) software were used. The degree of DNA fragmentation was assessed using the TUNEL method. To analyze the expression of miR-34a and miR-210, quantitative real-time PCR was performed using the miRCURY LNA SYBR Green PCR Kit («Qiagen» GmbH, Hilden, Germany) and the Rotor-Gene Q PCR product detection system («Qiagen» GmbH, Hilden, Germany).Results. The study of ejaculate using the Computer Assisted Sperm Analysis System (CASA) method revealed a statistically significant relationship between the level of DNA fragmentation of sperm and indicators of the speed of their movement: rectilinear (VSL) (r = -0.522726; p < 0.01), curvilinear (VCL) (r = -0.499096; p < 0.01), along the middle path (VAP) (r = -0.429533; p < 0.01), as well as with the amplitude of lateral head displacement (ALH) (r = -0.294779, p < 0.01), the linearity of their curvilinear path (LIN) (r = -0.385796; p < 0.01), the degree of straight-line movements (STR) (r = -0.268248; p < 0.05) and their progressive mobility (r = -0.411547; p < 0.01). A study of the level of microRNA expression in sperm exosomes revealed a statistically significant decrease in its miRNA-34a pool (p = 0.0116). According to the Chaddock scale, the strength of the correlation between miR-210 expression and the effectiveness of ART programs was moderate (0.437993). The inverse relationship between miR-34a expression and IVF and ICSI results was weak (0.135314).Conclusion. The analysis of exosomal microRNA-34a and microRNA-210, which are involved in the regulation of spermatogenesis, reveals a direct correlation between their variations and changes in the kinetic and morphological parameters of gametes. It also indicates a relationship with the state of DNA fragmentation. These findings suggest varying levels of gene expression among infertile patients, men with proven fertility, and those undergoing assisted reproductive technology (ART) treatments, both with successful and repeated unsuccessful outcomes.

Recent decline in sperm motility among donor candidates at a sperm bank in Denmark.

Has there been variation in semen quality among men applying to be sperm donors (i.e. donor candidates) in Denmark in recent years (2017-2022)? The motile sperm concentration and total motile sperm count (TMSC) in ejaculates-both measures of sperm quality-declined by as much as 22% from 2019 to 2022. Questions remain about whether human semen quality has declined in recent years. Whilst some studies provide evidence for a decline in human semen quality, these findings have been disputed owing to potential biases in the populations studied or in the methods used to measure semen quality. Resolution of this issue has important implications for human fertility, as well as for those involved in the recruitment of sperm donors for use in medically assisted reproduction. We obtained data on the semen quality of ejaculates previously collected from 2017 to 2022 at sperm bank locations in four cities in Denmark: Aarhus, Aalborg, Copenhagen, and Odense. Our study focuses on the single semen samples provided by 6758 donor candidates aged between 18 and 45 years old to determine whether their sperm quality met a minimum criterion for them to be accepted as sperm donors. All ejaculates were analyzed within 1 hour of production. Semen volume (ml) was estimated by weight and both the concentration (106/ml) of sperm as well as the concentration of motile sperm (World Health Organization grades a and b) were measured using the same protocols and computer-assisted semen analysis system across all years at each site. Statistical analyses of the semen variables were controlled for age and donation site, as well as the average monthly high temperature when the ejaculate was produced. From 2017 to 2019, semen volume, sperm concentration, and total sperm count in the ejaculates of donor candidates increased by 2-12%. Then, from 2019 to 2022, sperm concentration and total sperm count changed by 0.1-5% from year to year, but none of those changes were statistically significant. In contrast, both motile sperm concentration and TMSC declined significantly, by 16% and 22%, respectively, between 2019 and 2022. Thus, the concentration of motile sperm in donor candidates declined from 18.4 [95% CL: 17.0, 20.0] million/ml in 2019 to 15.5 [14.4, 16.7] million/ml in 2022, and TMSC declined from 61.4 [55.8, 67.5] million per ejaculate in 2019 to 48.1 [44.1, 52.4] million in 2022. We cannot determine from the available data the causes of the decline in semen quality of donor candidates from 2019 to 2022. However, as this period coincides with lockdowns and changes in work patterns during the coronavirus disease 2019 pandemic, it is possible that changes in motile sperm concentration and TMSC were the result of changes in the lifestyles of the men whose semen was analyzed. Men providing initial semen samples at sperm banks, when applying to be sperm donors, are a useful population in which to monitor changes in human semen quality over time. Our results have implications for human fertility and the recruitment of sperm donors for medically assisted reproduction, where motile sperm concentration is an essential selection criterion because it influences fertility. We suggest that gathering health and lifestyle data on donor candidates at sperm banks might help to identify causal factors for the decline of sperm quality that could be addressed and intervention, if desired, could be personalized for each accepted donor. No external funding was obtained for this study. E.L. and A.-B.S. are employees of Cryos International. AP reports paid consultancy for Cryos International, Cytoswim Ltd, Exceed Health, and Merck Serono in the last 2 years of this study, but all monies were paid to the University of Sheffield (former employer). AP is also an unpaid trustee of the Progress Educational Trust (Charity Number 1139856). RM declares support from Cryos International to present results of this research at ESHRE 2023. None of the authors were directly involved in the collection or physical analysis of semen samples. N/A.

Which is better to predict erectile dysfunction and male sexual function in the context of metabolic syndrome: triglyceride-glucose index or visceral adiposity index?: a retrospective cross-sectional study.

Many factors influence male sexual function, including metabolic disorders such as metabolic syndrome (MetS). We aimed to investigate the effects of two metabolic indices, the triglyceride-glucose (TyG) index and the visceral adiposity index (VAI), on male sexual function. A total of 400 men having sexual dysfunction were included. Anthropological data, comorbidities were recorded. Serum total testosterone, prolactin, and estradiol levels were recorded. Sex-specific VAI was calculated using the [(WC/39.68) + (1.88xMI)] × (TG/1.03) × (1.31/HDL) formula and using Ln (fasting triglycerides) × (fasting glucose)/2] formula, TyG index was calculated. Turkish-validated 15-item long-form of the International Index of Erectile Dysfunction (IIEF) questionnaire and male sexual health questionnaire (MSHQ) were used for erectile function and ejaculatory function, respectively. The ROC analysis was used to evaluate the predictive abilities of TyG and VAI cut-off values for ED risk. A higher TyG index and VAI were associated with an increased risk of ED. The presence of MetS further worsened sexual function, with lower scores in sexual satisfaction, orgasm, desire, and general satisfaction. The TyG index and VAI showed similar predictive abilities for ED. Patients with MetS had worse ejaculation quality compared to those without MetS. These findings highlight the potential of the TyG index and VAI as convenient tools for predicting and assessing sexual dysfunction in men, particularly in the context of metabolic disorders. Early detection and intervention for metabolic syndrome and insulin resistance may help to mitigate their negative impact on male sexual function.

The correlation between human seminal plasma sialoproteins and ejaculate parameters

Sialic acids are negatively charged carbohydrates that are components of saccharide chains covalently linked to macromolecules. Sialylated glycoproteins are important for most biological processes, including reproduction, where they are associated with spermatogenesis, sperm motility, immune responses, and fertilization. Changes in the glycoprotein profile or sialylation in glycoproteins are likely to affect the quality of ejaculate. The aim of this study was to determine differences in the degree of sialylation between normozoospermic ejaculates and ejaculates with a pathological spermiogram using two lectins, Sambucus nigra (SNA) and Maackia amurensis (MAL II/MAA) recognizing α-2,6 or α-2,3 linkage of Sia to galactosyl residues. Our results show a close relationship between seminal plasma (SP) sialoproteins and the presence of anti-sperm antibodies in the ejaculate, apoptotic spermatozoa, and ejaculate quality. Using mass spectrometry, we identified SP sialoproteins such as, semenogelins, glycodelin, prolactin-inducible protein, lactotransferrin, and clusterin that are associated with spermatozoa and contribute to the modulation of the immune response and sperm apoptosis. Our findings suggest a correlation between the degree of SP glycoprotein sialylation and the existence of possible pathological states of spermatozoa and reproductive organs. Glycoproteins sialylation represents a potential parameter reflecting the overall quality of ejaculate and could potentially be utilised in diagnostics.

Monitoring of taurine dietary supplementation effect on parameters of Duroc boar ejaculate in summer season.

The aim of this experiment was to find out whether the taurine supplementation in daily ration had an effect on quantity or quality of Duroc boar ejaculate. The experiment duration was from June to August, when it could assumed the possible occurrence of heat stress. For the study was chosen 12 Duroc boars of approximately the same age and condition. The control group of 6 Duroc boars was fed only by basic diet and the experimental group of 6 Duroc boars was fed by the same basic diet with supplementation of 15 g taurine/boar/day. Ejaculate was collected once a week by hand glowed technique. From ejaculate parameters were monitored volume of ejaculate, sperm concentration, total amount of sperm, morphologically abnormal sperm, taurine concentration and GSH/GSSH concentration. From microscopic analysis, results were statistically significant in motility in June and July (P<0.05). In biochemical results, a significant difference (P<0.05) has been found between the experimental groups in the concentrations of taurine as well as GSH/GSSG in ejaculate which indicates the effect of heat stress on boars during the experimental period.

Analysis of the relationship between the frequency of erythrocytes with micronuclei, semen quality and reproductive indicators in artificial insemination of foxes

Relevance. Reproductive biotechnologies are widely used in various branches of animal husbandry. Their scientifically based application shows high results in obtaining offspring. In fur-bearing cellular animal husbandry, reproductive biotechnology methods also give certain results. At the same time, in order to increase and predict the success in reproduction of fur-bearing animals, it is necessary to control the selection and selection of parental individuals, in particular male sperm donors during artificial insemination.Methods. The studies were performed in February — May 2023. The object of the study were ejaculate samples, peripheral blood smears of 16 silver-black fox males, as well as the corresponding results of artificial insemination (AI). To evaluate ejaculates, the following were studied: sperm concentration — by photometry, their percentage distribution by type of movement — on the ISAS system, sperm morphology — by microscopy. In peripheral blood smears, the frequency of occurrence of erythrocytes with micronuclei was determined using a micronucleus test (MTA). The results of the IO were evaluated by the number and percentage of pregnant and missing females, the number of live and dead puppies born.Results. The results showed that with the frequency of occurrence of erythrocytes with micronuclei above 2%, the sperm content in ejaculates was from 45 to 54.5% with non-accessible movement (type c), and from 22.7 to 44.2% stationary (type d). Also, abnormal sperm forms prevailed in the ejaculates of these males (63–83%), while 66–100% of missing females were noted. At the same time, males with MINTS of less than 1.5% demonstrated good ejaculate quality and high efficiency of artificial insemination.

FERTILITY FACTORS. D-HORMONE IN PREVENTION OF REPRODUCOPATHIES CAUSED BY BENIGN PROSTATIC HYPERPLASIA (Literature review)

In accordance with the statistical data, the spreading of benign prostatic hyperplasia (BPH) is increasing, because modern stressful lifestyle causes sex hormones disbalances and other dyshormonal conditions which lead to pathologies’ development at younger ages. The pathogenesis of BPH is connected with disturbances of regulation in the hypothalamo-hypophyseal system and with the changes of the regulatory signals intensity which go to the reproductive organs. The central pathogenic links of this pathology’ development are: the declining of efficacy of hypothalamus functioning regulation which carries out on the basis of feedback; the changes of the pituitary gland cells sensitivity to the releasing hormones of hypothalamus and disbalance of sex hormones. In addition to gonadotropins prolactin has a significant impact on the BPH development. Its hypersecretion causes the development of hyperplastic age-related changes in prostate gland. The introduction of dopamine receptors antagonists such as sulpiride causes the increasing of prolactin concentration and declining of gonadotropins releasing. “Sulpiride model” is broadly used in the experimental endocrinology for studying prostate-active drugs and is characterized by sex hormones disbalance' and inflammation processes’ development. The development of the pathology is linked with the disturbance of testosterone synthesis in the testicles, insufficiency of prostate gland functioning and increasing of prolactin and estradiol concentrations which provoke proliferative changes. The androgens disbalance is the main cause of BHP development, therefore, the “Testosterone model” based on testosterone’ and estradiol’ slow releasing implants is broadly used for modeling of urination disturbance in rats which imitates the same condition in elderly men with benign prostatic hyperplasia. The prostatitis, which developed under the condition of BHP, causes the disturbances of spermatogenesis during experimental modeling and in the clinical practice when spermatopathies, sexual disturbances and hypofertility are diagnosed in patients. The traditional BHP therapeutic methods are not always effective and the new therapeutic approaches of hypofertility caused by prostate gland pathologies are developed by investigators. Thus, there are data about correlation between increased vitamin D consumption and decreasing of BHP spreading, between gland volume and cells proliferation in patients with this pathology. Vitamin D level influences the quantity and quality of ejaculate. The deficiency of vitamin D is associated with lower proportion of sex hormones in young men and choriogonin sensitivity of Leydig's cells to be diminished. It has been experimentally shown that cholecalciferol regulates steroid hormones genesis, mitigates the age-related declining of testicles activity, protects the testicles in old animals and the unchanged spermatogenesis in normal males which received vitamin D3 can be considered to be the result of estrogens and androgens balance. The androgens receptors as well as vitamin D receptors have been found in prostate gland in health males and in those with hyperplasia. In addition to its classical function in calcium homeostasis and bones health, vitamin D inhibits tumor growth in different tissues including prostate gland. Vitamin D metabolites have been determined to be anti-inflammatory, proapoptotic and antiproliferative. D-hormone application potentiates the specific properties of prostate gland protectors, recovers reproductive capacity under the condition of experimental chronic prostatitis of inflammatory genesis as well as traumatic. All facts listed above allow to confirm the positive role of D-hormone in the regulation of prostate gland and reproductive function in patients with BHP and recovering of fertility under the condition of this pathology. The literature search for this review has been carried out on databases PubMed, Science Direct, Europa PMC, BMC, MedLine etc.

A retrospective analysis investigating the effects of Telazol® and medetomidine on ejaculate characteristics in cheetahs (Acinonyx jubatus)

Zoo managed cheetahs provide an insurance population for wild cheetahs that are under threat of extinction from habitat loss, lack of prey, competition, pet trade and poaching for skin and bones. Assisted reproductive techniques including artificial insemination, in vitro fertilization, and embryo transfer augment natural breeding programs but rely on good quality semen for best results. It is understood that anesthesia can affect semen characteristics such as ejaculate volume, total sperm count, sperm motility, and incidence of urine contamination. Thus, the aim of this study was to conduct a retrospective analysis of 23 years of data to investigate sperm parameters of semen collected under anesthesia using medetomidine in combination with butorphanol and midazolam or Telazol® alone. Electroejaculation records (Medetomidine, Butorphanol, and Midazolam anesthetized n = 59 ejaculates, from 30 cheetahs, Telazol® anesthetized, n= 169 ejaculates, from 72 cheetahs) were evaluated for incidence of urine contamination. Electroejaculation records (Medetomidine, Butorphanol, and Midazolam anesthetized n = 21 ejaculates, from 17 cheetahs, Telazol® anesthetized, n = 143 ejaculates, from 63 cheetahs) were evaluated for total sperm count, total motility, ejaculate volume, and testicle size. Telazol® treated cheetahs had a numerically higher total sperm count (Median ± SD: 42.58 ± 77.8 × 106 spermatozoa) compared to those treated with medetomidine (Median ± SD: 31.2 ±44.58 × 106 spermatozoa), and a significantly (p &amp;lt; 0.05) higher sperm motility (Median ± SD: 70.0 ± 9.71%) compared to medetomidine (Median ± SD: 53.0 ± 16.41%) treated cheetahs. The findings of this study indicate that medetomidine anesthesia results in significantly lower sperm motility and Telazol® anesthesia results in a higher total sperm count and motility, thus resulting in higher quality ejaculate. This information can aid in the veterinary management of the species when involved in genome resource banking and assisted reproductive technologies.

Features of reproductive capacity and state of prooxidant-antioxidant homeo-stasis in breeding boars of different breeds

Temperature stress is caused by the long-term effect of high or low average daily temperature, which leads to a decrease in the quality of sperm in fetuses and processes from the formation of the leading herd of pigs, which causes a reduction in the production of livestock products. Changes in lipid peroxidation accompany the inhibition of reproductive function in breeding boars of various breeds. The research method was to determine the features of pro-oxidant-antioxidant homeostasis in breeding boars of different species depending on the keeping conditions. For the experiment, six breeding boars of other breeds, similar in age, live weight, and quality of sperm production, were selected, and 2 study groups of 3 heads in the skin were formed. The first group consisted of animals of the large white breed, and the second group – was from Myrhorod. Animals were fed following feed regulations. The influence of heat stress on the quality of sperm production of breeding boars of the large white breed is manifested in a decrease in the following indicators: ejaculate volume – by 20.3 %, sperm motility – by 20.6 %, sperm concentration – by 28.3 %, the number of live sperm – by 54.6 %, heat resistance by 26.4 %. As the temperature rises, peroxidation processes accelerate – the content of TBC-active compounds in sperm increases by 1.3 times. Catalase activity in seminal plasma and sperm increases by 16.0 % and 21.0 %, respectively. An increase in indoor temperature harms the indicators of sperm production of breeding boars of the Myrhorod breed: ejaculate volume – by 18.6 %, sperm motility – by 10.5%, sperm concentration – by 30.3 %, number of live sperm – by 34.7 %, thermal resistance – by 23.5 %. The changes occur against the background of an increase in superoxide dismutase activity – 21.0 % (sperm) and 5.0 % (sperm plasma), a decrease in ascorbic acid saturation by 10.0 % and 52.2 %, respectively. The establishment of an interbreed difference in the quality of ejaculates in the direction of the superiority of indicators in animals of the large white breed compared to the Myrhorod breed on the 60th day of the primary period in the summer in terms of ejaculate volume (p = 0.001) and sperm concentration was established. However, according to the level of heat resistance of sperm, animals of the second breed were characterized by higher values. At the end of the 45th day of temperature stress, a probable predominance of ascorbic acid (p = 0.001) and dehydroascorbic acid (p = 0.01) content was observed in the semen of animals of the Myrhorod breed compared to large white.

Semen quality of rams fed a liposomal vitamin-mineral supplement during the period of sexual rest

To increase the sexual activity and sperm quality of rams during the period of sexual rest, it is necessary to increase the consumption rates of vitamins and microelements to the level of the mating season. The aim of the work was to find out the effect of feeding a liposomal vitamin-mineral supplement on the quality parameters of ram sperm during the period of sexual rest. The experiment was conducted on 12 clinically healthy Texel rams, aged 2–4 years, during the period of sexual rest (March–May). Animals were divided into two groups: control and experimental, 6 heads each. The control rams received the basic diet, the animals of the experimental group were individually added to the combined feed for 45 days with a feed additive in the form of a liposomal emulsion, which included: vitamins A, D3, E, C and zinc gluconate. After the end of supplement feeding, ejaculates were collected from the rams for research. The motility, kinetic indicators of sperm, the activity of succinate dehydrogenase (SDH) and cytochrome oxidase (CO) in sperm, as well as the activity of antioxidant protection enzymes superoxide dismutase (SOD), glutathione peroxidase (GPO) and catalase (CAT) were determined. It was established that feeding liposomal vitamin-mineral supplement to rams during the period of sexual rest significantly (P &lt; 0.05) increases sperm activity, their kinetic indicators: curvilinear (VCL), straight-line (VSL) and average speed (VAP) of sperm movement (P &lt; 0.05–0.001), the activity of enzymes – markers of the fertilizing capacity of SDH and CO sperm (P &lt; 0.05–0.001). At the same time, there was a significant (P &lt; 0.05) decrease in the activity of SOD with a simultaneous significant (by 23.3–25.0 %) increase in the activity of GPO and CAT (P &lt; 0.01). Higher indicators of the quality of ejaculates of rams under the influence of vitamins A, D3, E, C and zinc gluconate indicate the possibility of obtaining high-quality sperm from breeding rams during the period of sexual rest.

Receptivity and Remating Propensity in Female Spodoptera litura (Fabricius) after Mating with an Irradiated Male or Its F1 Male Progeny.

The 'Inherited or F1 sterility technique' (IS), using sub-sterilized male moths, is a widely proposed pest management tool for Lepidoptera pests in general, and the tobacco cutworm Spodoptera litura (Fabr.) in particular. However, the multiple mating tendency of female moths and the ejaculate quality of male moths might influence the efficiency of this technique. Reduced ejaculate quality was observed in irradiated males, as evidenced by radiation's impact on certain bio-parameters, such as the weight of the spermatophores and their protein content, sperm count, the molecular expression of the sex peptide receptor (SPR) and egg fertility, with a greater impact in F1 male progeny. During the remating of females with untreated males, irrespective of the irradiation status of the first male, there was an increase in calling behavior, remating propensity and fertility in females, with a larger time gap between consecutive matings. The ability of F1 male progeny to check remating propensity in females 24 h after the initial mating was lower than that of unirradiated males. Partially sterile (130 Gy) males were as successful as unirradiated males in inducing the level of mating refractoriness in females. Decreased ejaculate quality in F1 male progeny could be associated with increased female receptivity during remating. Understanding the influence of male moth irradiation, insemination quality and post (initial)-mating intervals on the remating behavior of normal female moths and induced sterility might help in simulation modeling and optimizing IS insect programs.

Impaired Testicular Function without Altering Testosterone Concentration Using an Anti-Follicular-Stimulating Hormone Receptor (Anti-FSHr) Single-Chain Variable Fragment (scFv) in Long-Tailed Macaques (Macaca fascicularis).

FSHr antibodies have been shown to inhibit the differentiation of spermatogonia to primary spermatocytes, resulting in infertility without a pathological effect on reproductive organs. The aim of this study was to develop single-chain variable fragments (scFvs) against the follicular-stimulating hormone receptor (anti-FSHr) using phage-display technology and to evaluate the effects of intratesticular administration of the anti-FSHr scFv on testicular function and testosterone production. A phage clone against the extracellular domain of FSHr selected from a scFv phagemid library was analyzed for binding kinetics by surface plasmon resonance. Using ultrasound guidance, three adult macaques (M. fascicularis) were administered with 1 mL of 0.4 mg/mL anti-FSHr scFv (treatment) and 1 mL sterile phosphate buffer solution (control) into the left and right rete testis, respectively. Testicular appearance and volume, ejaculate quality, and serum testosterone levels were recorded on day 0 (before injection) and on days 7, 28, and 56 (after injection). Testicular tissue biopsies were performed on day 7 and day 56 to quantify the mRNA expressions of androgen binding protein (ABP), inhibin subunit beta B (IHBB), and vascular endothelial growth factor A (VEGFA). The results demonstrated that the anti-FSHr scFv molecule was calculated as 27 kDa with a dissociation constant (KD) of 1.03 µM. The volume of the anti-FSHr scFv-injected testicle was reduced on days 28 and 56 compared with day 0 (p < 0.05). Total sperm number was reduced from day 0 (36.4 × 106 cells) to day 56 (1.6 × 106 cells) (p < 0.05). The percentage of sperm motility decreased from day 0 (81.7 ± 1.0%) to day 7 (23.3 ± 1.9%), day 28 (41.7 ± 53.4%), and day 56 (8.3 ± 1.9%) (p < 0.05). Sperm viability on day 0 was 86.8 ± 0.5%, which reduced to 64.2 ± 1.5%, 67.1 ± 2.2%, and 9.3 ± 1.1% on days 7, 28, and 56, respectively (p < 0.05). The expression of ABP and VEGFA on days 7 (14.2- and 3.2-fold) and 56 (5.6- and 5.5-fold) was less in the scFv-treated testicle compared with the controls (p < 0.05). On day 56, the expression of IHBB was less (p < 0.05) in the treated testis (1.3-fold) compared with the controls. Serum testosterone levels were unchanged throughout the study period (p > 0.05). This study characterized the anti-FSHr scFv and demonstrated that treatment with anti-FSHr ameliorates testicular function without altering testosterone levels, offering a potential alternative contraceptive for the long-tailed macaques.

Effects of Exogenous Human Chorionic Gonadotropin Administration on Plasma Testosterone and Semen Production in the Veiled Chameleon (Chamaeleo calyptratus)

Gamete collection, gamete preservation, hormone analysis, and artificial insemination have become integral parts of in situ and ex situ conservation programs for threatened and endangered species, and while these methods have been used to assist conservation in many different vertebrate groups, limited work has been done in reptiles. For example, semen collection in lizards has only recently been described and determined to be safe. The purpose of this study was to determine effective doses of human chorionic gonadotropin (hCG) for increasing plasma testosterone concentrations in the veiled chameleon (Chamaeleo calyptratus), as well as to determine whether the rise in testosterone impacts semen collection. A cross-over design was used. Chameleons were given injections of 100 IU, 200 IU, and 300 IU of hCG/animal and serial plasma testosterone measurements were collected over 24 hours. 100 IU/animal was determined to increase plasma testosterone concentrations similar to 200 IU, and 300 IU/animal. Following this, we determined weekly injections of hCG (100 IU/animal) would maintain elevated testosterone concentrations over 30 days. Additionally, we determined that elevated testosterone secondary to repeated injections of hCG decreased testicular size as determined by ultrasound. Repeated hCG injections and long-term elevation of plasma testosterone concentrations did not increase the likelihood to collect a semen sample with electroejaculation or improve ejaculate quality. Further research is needed to exogenously stimulate spermatogenesis and increase ejaculate quality to perform timed semen collections in male reptile species.

Age‐related changes in male ant sperm quality

AbstractSperm cells age during storage in the male reproductive organs, which can lead ejaculate quality to decline dramatically over time. Such may pose a particular concern in species with constrained sperm supplies, where adult males cannot replenish their sperm reserves. In eusocial hymenopterans (i.e. ants, bees and wasps with permanent castes), males typically reach adulthood with a limited quantity of sperm, and their testes deteriorate shortly thereafter. In adult life, both sexes mate during a single reproductive event. While males die quickly after copulation, they persist posthumously via sperm stored in their mates' spermatheca, sometimes for decades. As a result of this mating system, males should experience intense selection pressure to produce and transfer high‐quality sperm. Using the black garden ant (Lasius niger) as a model system, we investigated whether the duration of storage in the accessory testes before copulation affected sperm quality. We found that there was no impact of male age on sperm number or viability and that sperm DNA fragmentation decreased rather that increased with age. These results highlight the extent to which male ants, and possibly other taxa with constrained sperm supplies, have evolved extremely specialized strategies to ensure reproductive performance.

Differential distribution of 5-formylcytosine and 5-carboxylcytosine in human spermatogenic cells and spermatozoa

Background. The epigenome of gametes is formed under the control of the developmental programme and the influence of environmental factors. How cytosine oxidation patterns are formed and altered in human spermatogenesis remains obscure so far.&#x0D; The aim of the study was to assess 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC) patterns in human spermatogenic cells and spermatozoa.&#x0D; Materials and Methods. The study was performed on testicular biopsy samples of 10 azoospermic patients and ejaculate samples of 5 sperm donors and 8 patients from infertile couples. The microscope slides were prepared for further indirect immunofluorescence to detect 5fC and 5caC and FISH to determine spermatogenic cell ploidy. &#x0D; Results. 5fC and 5caC were undetectable in mitotic and meiotic chromosomes of spermatogenic cells, and was present exclusively in some spermatogonia and spermatid interphase nuclei as well as in some ejaculated spermatozoa. The frequency of spermatozoa with 5fC and 5caC varied in a wide range and was higher in patients than in sperm donors (p=0,007, p=0,028). The increase in frequency of spermatozoa with 5fC and 5caC was accompanied with the decrease in frequency of morphologically normal and progressively motile spermatozoa.&#x0D; Conclusions. 5fC and 5caC are differentially distributed in human spermatogenic cells and spermatozoa. The immunocytochemically detected increase of 5fC and 5caC in individual spermatozoa is most likely induced by oxidative stress caused by effects of internal and external factors rather than developmental programme. The evaluation of 5fC and 5caC in spermatozoa can be potentially used as an additional criterion of ejaculate quality.

Evaluation of semen quality in young men in Republic of North Macedonia

Research data show that in the last 50 years (1938-1991) there has been a trend of decreasing sperm concentration in the male population in Europe by 2.3% and in the USA by 0.8%. The reasons for such negative trend are not known, but it is assumed that lifestyle and environmental factors have an influence on genetic factors. Aim of this study was to evaluate sperm quality in young, healthy men in our country, and to compare sperm quality in our population with others in the world. Material and methods: Ejaculates from 203 healthy male subjects, aged 18-32, were stored in a thermostat at 36⁰C and analyzed manually on a native slide and hematoxylin-eosin-stained slides, under a phase contrast microscope. Sperm motility was assessed at two-time intervals, group A, 60 minutes after ejaculation and group B, 120 minutes after ejaculation, while sperm concentration and sperm morphology were assessed at one time interval. Results: Semen analysis showed an average volume of ejaculate 3.45 ± 1.5 ml, sperm concentration in 1 milliliter62.4 ± 39.2 x10(6) /ml, while total sperm concentration was 211.2 ± 173.2 x10(6). In group A, values for progressive spermatozoa were 48.6 ± 18.1 x10(6) /ml and in group B, values for progressive spermatozoa were 47.9 ± 17.3 x10(6) /ml. There was no statistically significant difference between the two time intervals (group A and group B) when interpreting sperm motility, p&gt;0.005. Analysis of morphology of spermatozoa showed a mean value of 6.9% for morphologically normal spermatozoa. Conclusion: The quality of ejaculate in young men in North Macedonia is in the range of reference values according to WHO, and also our results are similar to those from Germany, Turkey, Bulgaria, Faroe Islands.

Post-pubertal functional outcomes of one-stage anatomical reconstruction of the corpus spongiosum, bulbo-spongiosus muscle and dartos in 46 children with proximalhypospadias

Anatomical studies of hypospadias show failure of zipping-up of histologically normal urethral plate and corpus spongiosum. With the commonly utilized substitution urethroplasties for proximal hypospadias, a reconstructed urethra of just an "epithelial-lined tube" with no spongiosal support, is apt to long-term urinary and ejaculatory dysfunctions. We completed a one-stage anatomical reconstruction in children with proximal hypospadias whenever the ventral curvature could be reduced to <30° and evaluated the post-pubertal outcomes. This is a retrospective analysis of prospectively maintained data on one-stage anatomical repair of proximal hypospadias between 2003 and 2021. In children with proximal hypospadias, the corpus spongiosum, bulbo-spongiosus muscle (BSM), Bucks', and Dartos' layers of the shaft were anatomically re-aligned prior to assessing the ventral curvature visually. When the curvature was >30°, the urethral plate was divided at the glans for a 2-stage procedure, and those patients were excluded from the study. Otherwise, the anatomical repair was continued (this series). The Hypospadias Objective Scoring Evaluation (HOSE) and the Paediatric Penile Perception Score (PPPS) were used for post-pubertal assessment. Prospective records provided details of 105 patients with proximal hypospadias who had complete primary anatomical repair. The median age at surgery was 1.6 years, and 15.9 years at the post-pubertal assessment. Forty-one (39%) had complications that necessitated re-operations. Thirty-five (33.3%) patients had complications involving the urethra. For fistula and diverticula, eighteen cases required only one corrective procedure, while one required two. Other 16 patients required an average of 1.78 corrective operations for severe chordee and/or breakdown, with 7 requiring Bracka's 2-stage procedure. Fifty patients (47.6%) were over 14 years old; 46 (92.0%) had pubertal reviews and scoring, while four were lost to follow-up. The mean HOSE score was 14.8/16, and the mean PPPS score was 17.8/18. Five patients had residual curvature of >10°. 17 and 10 patients, respectively, were unable to comment on glans firmness and ejaculation quality. During erections, 26/29 (89.7%) patients reported a firm glans, and 36/36 (100%) reported normal ejaculations. This study proves the need for reconstruction of normal anatomy for normal post-pubertal function. In all proximal hypospadias, we strongly recommend anatomical reconstruction (zipping up) of the corpus spongiosum and BSM. When the curvature can be reduced to <30°, a complete one-stage reconstruction is possible; otherwise, anatomical reconstruction of the bulbar and proximal penile urethra is recommended, reducing the length of the epithelial-lined substitution tube for the distal shaft and glans.

Ranking parameters driving siring success during sperm competition in the North African houbara bustard

Sperm competition is a powerful force driving the evolution of ejaculate and sperm traits. However, the outcome of sperm competition depends on many traits that extend beyond ejaculate quality. Here, we study male North African houbara bustards (Chlamydotis undulata undulata) competing for egg fertilization, after artificial insemination, with the aim to rank the importance of 14 parameters as drivers of siring success. Using a machine learning approach, we show that traits independent of male quality (i.e., insemination order, delay between insemination and egg laying) are the most important predictors of siring success. Traits describing intrinsic male quality (i.e., number of sperm in the ejaculate, mass motility index) are also positively associated with siring success, but their contribution to explaining the outcome of sperm competition is much lower than for insemination order. Overall, this analysis shows that males mating at the last position in the mating sequence have the best chance to win the competition for egg fertilization. This raises the question of the importance of female behavior as determinant of mating order.