Number Of Spermatozoa Research Articles

Impact of Oxidative Stress on Sperm Quality in Oligozoospermia and Normozoospermia Males Without Obvious Causes of Infertility

Background: Male factors contribute to approximately 50% of infertile couples. However, obvious causes remain unknown in many cases. This observational study aimed to investigate the associations of clinical and lifestyle parameters with sperm parameters. Methods: This study enrolled 41 men in infertile couples without obvious causes for male infertility from July 2023 to April 2024. Semen samples were evaluated for sperm number, motility, DNA fragmentation, and oxidative stress (OS) marker oxidation–reduction potential (ORP). Blood samples were analyzed for biochemical parameters, including advanced glycation end products (AGEs), and systemic OS marker diacron-reactive oxygen metabolites (d-ROMs). Skin-accumulated AGE levels were identified with an autofluorescence method. Lifestyle factors were assessed with a lifestyle questionnaire. Results: Most of the participants were under 40 years old and non-obese with normal clinical parameters. Multiple regression analyses revealed that body mass index, serum d-ROMs, and semen ORP levels were independently associated with decreased sperm number. Additionally, serum zinc and semen ORP levels were associated with sperm motility. Furthermore, serum zinc and high-density lipoprotein cholesterol levels were associated with sperm progressive motility and DNA fragmentation, respectively. The rest of the clinical and lifestyle factors, including skin-accumulated and serum AGE levels, were not correlated with any sperm parameters. Furthermore, serum d-ROM and semen ORP levels were not correlated with each other or any of the clinical and lifestyle factors. Conclusions: Our present study indicates that both systemic and local OS may be independently involved in sperm abnormality in healthy men without obvious causes for male infertility.

The effects of modest intake of soy milk enriched with Lactobacillus casei and omega-3 on the testis parameters in diabetic rats: a stereological study.

Diabetes mellitus is a widely distributed endocrine disorder in the world. Altered reproductive function is a notable long-term consequence of type 1 diabetes mellitus (T1DM). In the current study, we assessed the effects of soya milk containing Lactobacillus casei and omega-3 on stereology of testes in type 1 diabetic rats. 30 male Sprague Dawley rats were randomly allocated into five groups. Streptozocin (STZ (60 mg/kg)) was applied for diabetes induction. The non-diabetic and diabetic control groups were fed with 1 ml of distilled water. Three treatment diabetic groups were fed 1 ml of Soy milk group (SM), Probiotic soy milk group (PSM), and Omega-3 probiotic soy milk group (OPSM) via intragastric gavage for 60 days. At the endpoint, the animals were sacrificed and serum luteinizing hormone (LH), Follicle-stimulating hormone (FSH), testosterone, MDA besides testicular, and seminal parameters were analyzed. The administration of soy milk supplemented with L. casei and omega-3 in diabetic rats elevated the concentrations of LH, FSH, testosterone, and reduced malondialdehyde (MDA). In addition, this combination improved sperm quality, enhanced the number of sperm with rapid progress, increased testis weight and volume, seminiferous tubule and germinal epithelium volume; and also augmented the number of spermatogonia, spermatocyte, round and long spermatids, Sertoli cells and Leydig cells. Supplementation with soy milk containing L. casei and omega-3 can inhibit T1DM-induced infertility rats through improving testicular parameters, enhancing sperm quality, and increasing Sertoli and Leydig cell number.

EP300‐interacting inhibitor of differentiation 3 is required for spermatogenesis in mice

AbstractBackgroundMammalian spermatogenesis is a highly complex process of cell proliferation, meiosis, and differentiation. A series of genes are expressed in an orderly and precise manner to ensure spermatogenesis, with chromatin undergoing intricate changes throughout. EP300‐interacting inhibitor of differentiation 3 (Eid3) is a testis‐enriched gene, but its role in male reproduction remains unclear.ObjectiveTo investigate the role of EID3 in male spermatogenesis and explore the potential underlying mechanism.Materials and MethodsWe generated Eid3 knockout mouse model using the CRISPR‐Cas9 system. We measured the expression of EID3 in mouse tissues and testicular cell populations by qRT‐PCR and western blot. Histological analysis, including hematoxylin and eosin (H&E) and periodic acid‐Schiff (PAS) staining, together with computer‐assisted sperm analysis (CASA), were performed to evaluate the effect of EID3 on spermatogenesis in mice. Light and ultrastructural microscopy were used to evaluate the morphology and structure of the Eid3−/− spermatozoa. We used western blot and immunofluorescence to further analyze the function of EID3 in spermiogenesis.ResultsEid3−/− mouse showed a significant decrease in sperm count, motility, and morphology. Loss of EID3 impaired the normal meiotic process and induced apoptosis of abnormally developing spermatocytes, ultimately resulting in the decrease of sperm cell number. Additionally, EID3 deficiency led to a decrease in histone acetylation levels in spermatids, impaired histone‐to‐protamine transition and chromatin condensation process, and ultimately resulted in abnormal sperm morphology.Discussion and ConclusionsThis study confirms for the first time that EID3 is crucial for meiosis and chromatin condensation during spermatogenesis, and EID3 deficiency leads to a significant decrease in sperm parameters. Given the high expression paradigm of Eid3 in human testis, EID3 likely plays a role in human reproduction. Future research could provide a new target for the clinical diagnosis and treatment of male infertility.

Prediction model of male reproductive function damage caused by CHOP chemotherapy regimen for non-Hodgkin's lymphoma

ObjectiveThe CHOP combined chemotherapy regimen (cyclophosphamide, doxorubicin, vincristine, and prednisone) is commonly used to treat non-Hodgkin Lymphoma (NHL). While these drugs are effective for cancer treatment, they may have side effects on the reproductive system that are poorly studied. This study used a mouse model to investigate the mechanisms of reproductive function impairment induced by the CHOP regimen and developed a predictive model for assessing reproductive damage with a non-invasive procedure.MethodsFrom 2022 to 2023, we statistically analyzed the changes of reproductive function of NHL patients before and after receiving CHOP regimen in the First Affiliated Hospital of Xiamen University. The NHL mouse model was established and divided into CHOP treatment group and control group. The weight of testis and epididymis, sperm quality and motility were compared between the two groups. Histopathological examination of testicular tissue was performed to determine pathological changes. ELISA was used to measure the expression of cytokines and cytokine pathways in serum, protein expression was analyzed by immunohistochemistry, and protein and mRNA levels of cytokines and pathways were evaluated by Western blotting and qPCR. Using stepwise regression method to select important factors, a prediction model of reproductive system damage was constructed.ResultsFifty-two NHL patients included in the questionnaire showed significant reproductive system damage after CHOP regimen treatment. The weight of testis and epididymis, as well as the number and vitality of sperm in the mouse model treatment group were significantly lower than those in the control group. Serum LH, FSH, estradiol and progesterone levels decreased significantly, while inhibin B levels increased significantly. There was no significant change in testosterone or prolactin levels. Inflammatory markers such as CSF-1, IL-1, IL-6, TGF-β1 and GDNF increased significantly, while the level of SOD1 decreased significantly. Immunohistochemical staining analysis showed that CAMP, Caspase3, CSF-1, GDNF, IL-1, IL-6, PRKACB, TGF-β1 and TXNDC5 were all expressed in spermatocytes, and the expression of therapeutic histones was significantly higher than that of the control group. Western blot analysis further detected the protein expression, and QPCR detected the mRNA content. The results showed that the expression of histone and mRNA in the treatment group was significantly higher than that in the control group. Stepwise regression method determined that estradiol (E2) was the most important variable in the prediction model, and the AUC for predicting reproductive damage was 1.ConclusionsThe CHOP regimen induces male reproductive toxicity, potentially mediated through alterations in hormone levels and increased expression of inflammatory cytokines and oxidative stress. Using E2 as the sole predictor in the model accurately predicts the extent of reproductive damage, offering a non-invasive method for detecting reproductive system damage.

Sexual size dimorphism affecting mate choice and reproduction in the pine sawyer beetle, Monochamus alternatus.

Monochamus alternatus is a notorious pest of the pine forests across East Asian countries. Both large and small individuals of either sex coexist in the wild. However, whether and how body size affects sexual selection outcomes and longevity remains unknown. In this study, we allowed a male or a female of small or large size to choose between two mates of varying sizes, which were from a wild-caught population. Our results revealed that mating latency and frequency of mating attempts varied across different social environments. Both large males and large females preferred larger mates, whereas this preference was not observed for the small males and small females. Male-male competition had no significant effect on ejaculation duration and frequency or sperm number in ejaculates. When a single male had an opportunity to choose between two females of different sizes, the reproductive investment of small males towards the small females was almost doubled compared to that of the large males, i.e., longer ejaculation duration, higher ejaculation frequency, and greater numbers of sperm transferred. However, the greater mating efforts of small males did not translate to greater fecundity/fertility of small females. We also showed that regardless of their body size, females always had greater fertility when they mated with large males but evidence for a trade-off between fecundity and longevity was not found. Alternatively, female fecundity was positively associated with longevity, and fertility was negatively associated with longevity and fecundity. We discussed these results placing it in a broader context of mate choice evolutionary dynamics.

Disperse Red 1 azo dye: Consequences of low-dose/low-concentration exposures in mice and zebrafish

Color Index Disperse Red 1 (DR1), an azo dye widely used in the textile industry and released into aquatic environments, is genotoxic in somatic cells, but little is known concerning its effects on the reproductive system or the early stages of embryonic development. We have assessed the effects on the spermatozoa of male mice following oral exposure to the dye, at low doses, for 14 days. Measured endpoints were DNA damage (comet assay), miRNA-34c levels, and sperm number, morphology, and motility. Exposure caused decreased miRNA-34c levels. We have also examined dye effects on zebrafish embryos and larvae, which included developmental impairment, altered glutathione transferase activity, and effects on reactive oxygen species and lipid peroxidation levels.

Ethnopharmacological insights: hyperoside from Marsdenia latifolia (Benth.) K.Schum. mitigates reproductive toxicity in male Wistar rats induced by manganese exposure

This study provides novel findings on the ameliorative effect of hyperoside isolated from Marsdenia latifolia leaves (HIGLL) in reproductive health challenged by manganese toxicity. The study investigated the efficacy of HIGLL on male fertility in rats exposed to manganese chloride (MnCl2). The rats received either MnCl2 (30 mg/L) or in combination with HIGLL (100 mg/kg or 200 mg/kg). MnCl2 reduced fluid intake, organ-body weight, body weight; sperm count, sperm viability, sperm density, sperm motility, semen viscosity, daily sperm production, testicular sperm number; testosterone, follicle-stimulating and luteinising hormones, oestradiol, testosterone/oestradiol ratio; G6PDH, ALP, glucose, NO, acid phosphatase, sialic acid, 17-β-HSD, superoxide dismutase, CAT, GST, GSH, and T-SH without modifying the semen pH and volume, while it correspondingly raised the abnormalities associated with morphology of sperm cells in head, neck, and tail; H2O2 and lipid peroxidation levels. HIGLL abrogated the damaged histoarchitecture of the testes and epididymis caused by MnCl2. HIGLL can serve as a therapeutic agent in the management of male reproductive disorders related to oxidative stress and endocrine disruption.

Selenium efficiency in protecting sperm quality and testicular parameters of South African indigenous Zulu rams exposed to heat stress.

The study investigated selenium's (Se) efficiency in preserving South African Zulu rams' sperm quality and testicular parameters when they were exposed to heat stress. Indigenous Zulu rams (20) between 2 and 5 years old were allocated into four groups, namely the Se, testicular heat stress (THS), selenium plus testicular heat stress (SeTHS), and control. Each group comprised five rams; the groups were balanced according to the rams' body weight and scrotal circumference. The Se and SeTHS groups received sodium selenite orally bi-weekly for 5 months. To induce heat stress, testicular heat insulation bags were wrapped around the testes of the rams receiving the THS and SeTHS treatments for 49 days. Semen was collected from the rams weekly from the third month onward; the first two months were for Se & THS acclimatization. In addition, testicular measurements were taken bi-weekly. Analysis of variance was used to analyze the sperm quality data. Duncan's multiple range test was used to compare the groups' data for significant differences. The results showed that the Se-supplemented rams' scrotal circumference was smaller (p < 0.05) compared with the other groups. The Se, SeTHS, and control groups demonstrated similar total sperm motility; in contrast, the THS and SeTHS groups recorded low and high total sperm motility, respectively, compared with other treatment groups (p < 0.05). The semen from the rams that received THS without Se displayed a significantly higher number of immotile sperm cells (p < 0.05) and poor sperm quality, including total and progressive motility, and kinematic parameters when compared with other treatments, suggesting that Se protects sperm against THS. We concluded that selenium protected some sperm parameters (TSM, PSM, MV, VCL, VSL) of THS- treated rams while others did not improve (RV, NSM, C, STR).

Normal male fertility in a mouse model of KPNA2 deficiency.

The nuclear transport of proteins is mediated by karyopherins and has been implicated to be crucial for germ cell and embryonic development. Deletion of distinct members of the karyopherin alpha family has been shown to cause male and female infertility in mice. Using a genetrap approach, we established mice deficient for KPNA2 (KPNA2 KO) and investigated the role of this protein in male germ cell development and fertility. Breeding of male KPNA2 KO mice leads to healthy offsprings in all cases albeit the absence of KPNA2 resulted in a reduction in sperm number by 60%. Analyses of the KPNA2 expression in wild-type mice revealed a strong KPNA2 presence in meiotic germ cells of all stages while a rapid decline is found in round spermatids. The high KPNA2 expression throughout all meiotic stages of sperm development suggests a possible function of KPNA2 during this phase, hence in its absence the spermatogenesis is not completely blocked. In KPNA2 KO mice, a higher portion of sperms presented with morphological abnormalities in the head and neck region, but a severe spermiogenesis defect was not found. Thus, we conclude that the function of KPNA2 in round spermatids is dispensable, as our mice do not show any signs of infertility. Our data provide evidence that KPNA2 is not crucial for male germ cell development and fertility.

Factors determining semen sample collection and semen quality parameters in African penguins Spheniscus demersus

Our research focuses on semen collection from 42 African penguin males, grouped by age, habituation levels, and reproductive season timing. We assess behavioral and physiological responses to dorso-abdominal massage, evaluating species-specific seminal traits using conventional and advanced methods. Positive behavioral responses corresponded with successful semen collection. Young and human-reared males exhibited more favorable behaviors, and samples containing spermatozoa were more likely collected during the reproductive season. Age did not influence sample collectability or spermatozoa presence, though mature males produced higher semen volumes. Young males exhibited more sperm morphological abnormalities, especially outside the breeding season. Sperm concentration and number per ejaculate showed no significant differences across age or seasonal groups. Young males had higher motile and progressive sperm percentages, while mature males had more static sperm. Additionally, percentages of live sperm and intact acrosomes were higher in mature males. Young males and samples from non-reproductive season presented more dead sperm with damaged acrosomes. Plasma membrane integrity positively correlated with age for live sperm and negatively for dying sperm. This research demonstrates the feasibility of semen collection from African penguins regardless of habituation level, fills the knowledge gap by describing sperm morphological abnormalities, and provides insights into using flow cytometry in Sphenisciformes.

Triazophos-induced spermotoxicity in rats: Protective effects of nano-quercetin

This study aimed to evaluate the spermotoxic potential of triazophos in rats and to check the possible shielding effect of quercetin and nano-quercetin against triazophos-induced toxicity. Rats in Group I were given olive oil as a vehicle. Group II and Group III received high-dose and low-dose triazophos, respectively. Oral administration of quercetin (Group IV) and nano-quercetin (Group VI) at a dose of 50 mg/kg body weight was given to two additional groups of animals. Two other high-dose triazophos groups were co-administered with quercetin (Group V) and nano-quercetin (Group VII).Triazophos administration for 60 days in rats altered the structural and functional parameters of spermatozoa and brought about a decline in total sperm count, percentage of viable sperms, drop in sperm motility, and decrease in the number of sperms showing normal morphology. It also decreased the number of spermatozoa with intact acrosomes and HOST-positive spermatozoa. Further, triazophos increased the levels of reactive oxygen species and triggered apoptotic pathways in spermatozoa in a dose-dependent manner. It decreased daily sperm production and caused histomorphological aberrations in the epididymis and vas deferens. Co-administration of nano-quercetin with triazophos effectively counteracted sperm-related pathological changes. Nano-quercetin offered better protection over quercetin in ameliorating the triazophos-induced spermotoxicity in rats.

Sperm collection in the domestic cat: A comparison of two techniques

Semen collection in cats in the clinic setting can be difficult. However, semen analysis is vital when evaluating breeding soundness of a male. Electroejaculation (EEJ) is currently the most reliable semen collection method but requires specialized equipment and training of the operator. Chemical ejaculation followed by urethral catheterization (UC) is a technique that allows semen collection without special equipment: a catheter is placed into the urethra of a sedated tom and semen is collected passively into the catheter. Earlier studies used the sedative medetomidine at high doses for this procedure. However, medetomidine has been replaced with dexmedetomidine in some countries. This study sought to compare the results of EEJ and UC for semen collection in the domestic cat using dexmedetomidine, a potent α2-adrenoceptor agonist (α2A), as a substitute for medetomidine at the equivalent dose to that used in earlier studies. Twelve domestic cats were collected thrice at weekly intervals. All cats received intramuscular ketamine (5 mg/kg) and intramuscular dexmedetomidine (30 μg/kg) for initial cleanout via EEJ, then randomly underwent either EEJ or UC one week apart. The EEJ was performed under the same anesthetic protocol as the initial cleanout. The UC was performed using intramuscular dexmedetomidine at a dose of 60 μg/kg. Success of collection, total sperm number, sperm morphology, and motility characteristics were analyzed. Sperm was collected successfully from all 12 cats via EEJ and from 11/12 via UC. There were no significant differences in the percentage of total motile, progressively motile, or morphologically normal sperm between ejaculate types when averaged across all cats or individual cats. Although UC yielded a lower volume and higher concentration ejaculate, it consistently produced a lower total sperm number than ejaculates retrieved via EEJ (17.91 x 106 total sperm for UC versus 46.51 x 106 total sperm for EEJ). These results indicated that dexmedetomidine is a very effective sedative and performed satisfactorily in both procedures at the doses used in this study. It was also safe with no adverse effects on healthy toms. EEJ remained the most reliable in terms of assessing semen quality and retrieving semen with adequate number of sperm for breeding purposes. However, UC with dexmedetomidine at this dose demonstrated a 92 % success rate, presenting itself as a remarkably consistent alternative.

Effect of refreezing extender on stallion sperm quality and embryo production after intracytoplasmic sperm injection

Intracytoplasmic sperm injection (ICSI) is a valuable assisted reproduction technology in clinical practice, especially when semen availability is limited. Since the number of sperm required per ICSI cycle is much less than the number of sperm available in a standard straw of frozen semen, refreezing semen at lower sperm concentrations could yield multiple straws for ICSI use. However, there is little data on the effect of sperm refreezing on ICSI outcomes, especially on the effect of extender used for refreezing. The objective of the present study was to evaluate the effect of refreezing extender on stallion sperm quality and embryo production after ICSI. Semen was frozen in a low egg yolk/glycerol/amide extender (Extender 1), then thawed, re-extended, and refrozen in each of three extenders. When sperm were refrozen in Extender 1, the cleavage rate was lower (P < 0.05) and the blastocyst development rate tended to be lower (P = 0.06) than for the once-frozen sperm. In contrast, when sperm were refrozen in high egg yolk/glycerol (Extender 2) or low egg yolk/milk/glycerol (Extender 3) extenders, cleavage and blastocyst development rates did not differ significantly from those for the once-frozen semen. Notably, sperm refrozen in Extender 1, which yielded the lowest ICSI outcomes, showed the highest sperm motility and viability, demonstrating that traditional measures of sperm quality were inadequate to assess the suitability of refrozen sperm for ICSI. In a follow-up experiment conducted to evaluate the effects of Extenders 1 and 3 when used for once-frozen semen, cleavage and blastocyst rates did not differ between extenders. In conclusion, the extender used to initially freeze stallion sperm may not significantly affect ICSI outcomes; however, the extender used for refreezing can significantly affect embryo production. Refrozen stallion semen can be effectively used for ICSI when low egg yolk/milk/glycerol extender is used for refreezing. Until further research is available, use of extenders without amides is recommended when refreezing stallion semen for ICSI.

9325 Obesity Alters Pomc And Kisspeptin Neuron Crosstalk Leading To Lower Luteinizing Hormone

Abstract Disclosure: P. Villa: None. R. Ruggiero-Ruff: None. B. Jamieson: None. R. Campbell: None. D. Coss: None. Obesity is a significant public health concern due to its high prevalence and numerous comorbidities. It is associated with hypogonadism in males, characterized by low testosterone and sperm number. Previous studies determined that these stem from dysregulation of hypothalamic circuitry that regulates reproduction, by unknown mechanisms, since studies are confounded by chronic nature of this condition that leads to synaptic changes and alterations in neuron responsiveness to stimuli. Herein, we used mice fed chronic high-fat diet, that mimics human obesity, to determine mechanisms of impairment at the level of the hypothalamus, in particular gonadotropin-releasing hormone (GnRH) neurons that regulate luteinizing hormone levels (LH), which in turn regulates testosterone synthesis. Consistent with obese humans, we demonstrated lower LH, and lower pulse frequency of LH secretion, but unchanged pituitary responsiveness to GnRH. Pulse frequency of LH is regulated by pulsatile GnRH secretion, which is controlled by kisspeptin. Peripheral and central kisspeptin injections, and DREADD-mediated activation of kisspeptin neurons, demonstrated that kisspeptin neurons were suppressed in obese mice. Thus, we investigated regulators of kisspeptin secretion. We determined that LH response to NMDA was lower in obese mice, corresponding to fewer glutamate receptors in kisspeptin neurons, which may be critical for kisspeptin synchronization. Given that kisspeptin neurons also interact with POMC neurons, which regulate satiety and are particularly affected by obesity, we examined their crosstalk, and determined that LH response to either DREADD-mediated activation of POMC neurons or central injection of αMSH, a product of POMC, is abolished in obese mice. This was accompanied by diminished levels of αMSH receptor, MC4R, in kisspeptin neurons. The reason may be that chronically higher activity of POMC neurons in response to obesity, downregulates αMSH receptors on target neurons, including kisspeptin. This may lead to suppression of kisspeptin neurons, and their inability to regulate pulsatile secretion of GnRH. Together, our studies determined that obesity leads to downregulation of receptors that regulate kisspeptin neurons, which is associated with lower LH pulse frequency, leading to lower LH and hypogonadism. Presentation: 6/2/2024

Media viscosity affects post-thaw ram sperm rheotactic behaviour.

The aim of this experiment was to assess the effect of media viscosity on ram sperm motility, kinematics and rheotaxis in vitro by using methylcellulose as a media thickener. Frozen-thawed semen of three rams was thawed and diluted in Tyrode's albumin lactate pyruvate (TALP) media supplemented with 0%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6% and 0.7% w/v of methylcellulose. Sperm motility and kinematic characteristics were analysed using computer-assisted sperm analysis (CASA). The rheotactic behaviour was assessed in a microfluidic channel, and the number of spermatozoa that passed the 10 mm point of a microfluidic channel over a 2min period against a flow rate of 30 μm/sec was assessed. The use of media with higher viscosity (higher levels of methylcellulose) resulted in significantly lower (p < .05) sperm motility and kinematic parameters. Moreover, higher levels of methylcellulose reduced (p < .05) the number of spermatozoa that exhibited positive rheotaxis. In conclusion, viscosity affected the kinematic properties and rheotactic behaviour of ram sperm.

No impact of a positive Chlamydia trachomatis serology on live-birth rate after intra-uterine insemination

ObjectiveTo study the impact of a CT serology on intrauterine insemination (IUI) cumulative live-birth rate (cLBR) in women with documented bilateral tubal patency. DesignCohort study SubjectsInfertile women with documented bilateral tubal patency and medical indication of IUI matched on the following criteria: woman's age, number of cycles completed and number of motile sperm inseminated (NMSI). ExposureThis retrospective, observational and monocentric cohort study compared women with positive CT serology matched 1:1 to control women with negative CT serology. Main outcome measuresCumulative LBR, rates of clinical pregnancy, spontaneous abortion, biochemical pregnancy. ResultsA total of 71 women in the CT positive group were matched to 71 women in the negative CT group, leading to compare 136 cycles per group. No statistically significant difference was observed between groups regarding the demographic and medical characteristics of couples. Cumulative LBR per woman was similar in both groups with 32.4% (n = 23) in the negative serology group Vs 25.4% (n = 18) in the positive CT group (NS). The rates of clinical pregnancy, spontaneous abortion, biochemical pregnancy were comparable between the two groups. ConclusionIn a population of infertile women with patent tubes, our study suggests that the serological status for CT has no impact on the IIU cLBR.

Freezability of Semen Collected from Ganders of 12 Breeds Covered by the Polish Genetic Resources Conservation Program

Abstract A rapid decline in wild animal biodiversity, as well as in the number of local livestock breeds of lower economical values, makes species and breeds protection and creation of genetic reserves a necessity and a challenge of the 21st century. For birds, semen freezing is still the best method of gene preservation ex situ in vitro. The aim of the presented study was to assess the susceptibility to freezing process and to establish the semen gene bank of twelve goose breeds covered by the Polish genetic resources conservation program. Sexually mature ganders were randomly selected from four northern local breeds: Kartuska, Pomorska, Rypinska, Suwalska, four southern local breeds: Garbonosa, Kielecka, Lubelska, Podkarpacka, and four foreign goose breeds: Kuban, Landes, Roman and Slovakian. Each breed was represented by 12 males, twelve repetitions were carried out. Semen samples were frozen in a computer-controlled cryogenic chamber using a method developed for White Koluda goose. The basic evaluation criterion was sperm morphology in fresh and freeze-thawed semen assessed in nigrosine-eosin stained histological smears. The percentage of live in total and live normal sperm in the fresh semen differed significantly (P&lt;0.05) between breeds. Semen of Kartuska and Suwalska ganders (northern breeds), contained the highest percent of live sperm – 94.4 and 94.1 respectively, including 52.5% and 54.2% of live normal sperm, while the lowest number of these forms was observed for Slovakian goose – 86.4% and 29.2%, respectively. The freezing process resulted in a significant (P&lt;0.05) reduction in the total number of live and live normal sperm and an increase in dead sperm in all groups. Cryopreservation process was the most effective in case of Suwalska goose semen (28.4% of live normal cells in thawed semen) and the less profitable for Landes (7.4% of live normal cells). The analyses showed significant differences between breeds in sperm morphology in fresh and thawed semen. Moreover, the high proportion of live spermatozoa in the fresh semen did not guarantee a high proportion of spermatozoa after thawing, indicating the differences in semen susceptibility to cryopreservation process depending on goose breed.

Gone but not forgotten: dynamics of sperm storage and potential ejaculate digestion in the black soldier fly Hermetia illucens.

Understanding the dynamics of sperm storage is essential to unravel the complexity of post-copulatory sexual selection processes in internally fertilized species. This physiological process goes from sperm transfer during copulation to its use for fertilization. In this context, the spatiotemporal dynamics of sperm storage were described in the black soldier fly (BSF) with fluorescence and transmission electron microscopy (TEM). BSF females have compartmentalized spermathecae with a transfer compartment, the fishnet canals, and a storage compartment, the reservoirs. Spermatozoa were counted both during and after mating in the two compartments. In addition to seminal fluids, the male transfers a mass of sperm in the fishnet canals, then only 49% of the transferred spermatozoa reach the reservoirs over two days. TEM observations of the fishnet canals revealed potential digestive functions, explaining the decline in the number and viability of spermatozoa in this compartment but not in the reservoirs. After one mating, females laid up to three fertile clutches, showing no constraints on sperm quantity or quality. Spermatic and ultrastructural investigations strongly suggest that BSF ejaculate acts both as a sperm plug and as a nuptial gift, reinforcing the interest in studying this farming insect as a new model for sexual selection.

Sauger testicular harvest eliminates sperm limitation during large‐scale saugeye (Walleye × Sauger) production

AbstractObjectivePoor milt yield via strip‐spawning in male Sauger Sander canadensis limits the production of saugeye (female Walleye S. vitreus × male Sauger) by the Ohio Division of Wildlife. Compared to strip‐spawning, collecting sperm via testicular harvest offers several advantages that increase production efficiency through enhanced sperm yield (e.g., requires fewer broodfish, allows higher sperm application rates, and increases fertilization rates). The benefits of testicular harvest have been demonstrated experimentally in Sauger but not at a production scale. In this study, I validated the large‐scale efficacy of testicular harvest in Sauger for saugeye production.MethodsI collected sperm from male Sauger by using either testicular harvest (42 fish) or strip‐spawning (222 fish), diluted samples with extender, and combined samples to create 9‐mL sperm pools (n = 21 for testicular harvest; n = 39 for strip‐spawning). I compared sperm quantity, sperm motility, and production attributes (broodstock requirements, sperm application rates, and expected fertilization) between testicular harvest and strip‐spawning collection methods.ResultBoth collection methods resulted in high‐quality sperm. Testicular harvest doubled the sperm yield (total number of sperm) per 9‐mL pool despite using only 2 fish/pool, while strip‐spawning required 5–7 fish/pool. Through this enhanced yield, testicular harvest produced enough sperm to fertilize 317 L of eggs (1.06 mL/L) by using 80% fewer males than strip‐spawning. Further, sperm application increased fourfold using testicular harvest (4.23 mL/L) while still requiring 30–40% fewer fish than are currently used to meet production. Adopting testicular harvest for saugeye production in 2023 increased broodstock sperm yield, allowed greater sperm application rates, reduced Sauger broodstock needs, and increased saugeye hatching rates (15–20% increase).ConclusionTesticular harvest provides an efficient approach for sperm collection during large‐scale production in fish species that produce small quantities of sperm.

Comparison of different processed products of Allium tuberosum Rottler for the treatment of mice asthenozoospermia.

Allium tuberosum Rottler improves sexual function and is used in the treatment of impotence and spermatorrhea. However, its chemical composition and mechanism of action remain unclear. This study investigates the chemical composition and mechanism of action of Allium tuberosum Rottler co-processed with salt and wine (GZP) in modulating testicular mitochondrial autophagy for the treatment of asthenozoospermia in mice. Adenine gavage + cyclophosphamide intraperitoneal injection was used to establish the model of asthenozoospermia, and six Allium tuberosum Rottler processed products were compared in the pharmacological efficacy for the treatment of asthenozoospermia in mice. The liquid chromatograph mass spectrometer (LC-MS) assay was performed to analyse the compositional changes in the GZP. The mechanism of GZP in the treatment of asthenozoospermia in mice was further investigated. The mitophagy was detected by transmission electron microscope (TEM) and immunofluorescence, respectively. Reactive oxygen species (ROS) were detected by probe. Protein expression was determined by Western blotting. GZP exhibited optimal therapeutic effects on asthenozoospermia in mice. It showed the best therapeutic effect in improving the total number of spermatozoa, sperm survival rate, improving sperm viability and reducing sperm deformity rate, alleviating the abnormal pathological morphology of mice testis, and increasing the serum testosterone (T), follicle-stimulating hormone (FSH) and prolactin (PRL) levels in mice. The LC-MS detection found that Allicin showed the most significant increase in GZP. Besides, GZP reduced ROS level and inhibited mitophagy in mice testicular tissues. Meanwhile, it restrained the expression of PINK1, Parkin, Light chain 3II (LC3-II)/Light chain 3I (LC3-I) and Caspase-3 proteins. GZP improves asthenozoospermia via inhibiting excessive mitophagy and protects the integrity of mitochondria by blocking the PINK1/Parkin signaling pathway. During which, the Allicin may play an important role.