Search

ObjectiveEvaluate reduction in progressively motile sperm per high power field (HPF) in midcycle cervical mucus after intercourse with Ovaprene: an investigational monthly non-hormonal vaginal contraceptive consisting of a vaginal ring and mechanical barrier, releasing spermiostatic ferrous gluconate.Study designOpen-label, multicenter study enrolling heterosexually-active women with previous permanent contraception. Participants underwent a baseline postcoital test cycle with no device to confirm the presence of sperm, followed by one diaphragm postcoital test cycle, one Ovaprene safety cycle, and two Ovaprene postcoital test cycles. In each postcoital test cycle, participants underwent a midcycle cervical mucus evaluation to confirm an Insler score ≥10 and absence of sperm, and then returned two to four hours after vaginal intercourse for repeat cervical mucus evaluation. We considered

The ex situ population of the endangered black-footed ferret (Mustela nigripes) has been experiencing declines in reproductive success over the past 30 years of human-managed care. A potential cause may be environmental-dependent inbreeding depression with diet being one of the contributing factors since ferrets are not fed their natural diet of prairie dogs. Here, we generated and analyzed semen proteome and transcriptome data from both wild and ex situ ferrets maintained on various diets. We identified 1757 proteins across all samples, with 149 proteins unique to the semen of wild ferrets and forming a ribosomal predicted protein-protein interaction cluster. Wild ferrets also differed from ex situ ferrets in their transcriptomic profile, showing enrichment in ribosomal RNA processing and potassium ion transport. Successful fertility outcomes documented for ex situ ferrets showed the strongest association with the semen transcriptome, with enrichment in genes involved in translation initiation and focal adhesion. Fertility also synergized with the effect of diet on differentially expressed transcriptomes, mainly affecting genes enriched in mitochondrial function. Our data and functional networks are important for understanding the causes and mechanisms of declining fertility in the ex situ ferret population and can be used as a resource for future conservation efforts.

CRISPR-Cas9 short guide RNA (sgRNA) library screening is a powerful approach to understand the molecular mechanisms of biological phenomena. However, its in vivo application is currently limited. Here, we developed our previously established in vitro revival screening method into an in vivo one to identify factors involved in spermatogenesis integrity by utilizing sperm capacitation as an indicator. By introducing an sgRNA library into testicular cells, we successfully pinpointed the retinal degeneration 3 (Rd3) gene as a significant factor in spermatogenesis. Single-cell RNA sequencing (scRNA-seq) analysis highlighted the high expression of Rd3 in round spermatids, and proteomics analysis indicated that Rd3 interacts with mitochondria. To search for cell-type-specific signaling pathways based on scRNA-seq and proteomics analyses, we developed a computational tool, Hub-Explorer. Through this, we discovered that Rd3 modulates oxidative stress by regulating mitochondrial distribution upon ciliogenesis induction. Collectively, our screening system provides a valuable in vivo approach to decipher molecular mechanisms in biological processes.

Aminergic signaling is known to play a critical role in regulating female reproductive processes in both mammals and insects. In Drosophila, the ortholog of noradrenaline, octopamine, is required for ovulation as well as several other female reproductive processes. Two octopamine receptors have already been shown to be expressed in the Drosophila reproductive tract and to be required for egg-laying: OAMB and Octβ2R. The Drosophila genome contains 4 additional octopamine receptors-Octα2R, Octβ1R, Octβ3R, and Oct-TyrR-but their cellular patterns of expression in the reproductive tract and potential contribution(s) to egg-laying are not known. In addition, the mechanisms by which OAMB and Octβ2R regulate reproduction are incompletely understood. Using a panel of MiMIC Gal4 lines, we show that Octα2R, Octβ1R, Octβ3R, and Oct-TyrR receptors are not detectable in either epithelium or muscle but are clearly expressed in neurons within the female fly reproductive tract. Optogenetic activation of neurons that express at least 3 types of octopamine receptors stimulates contractions in the lateral oviduct. We also find that octopamine stimulates calcium transients in the sperm storage organs and that its effects in spermathecal, secretory cells, can be blocked by knock-down of OAMB. These data extend our understanding of the pathways by which octopamine regulates egg-laying in Drosophila and raise the possibility that multiple octopamine receptor subtypes could play a role in this process.

There has recently been an explosion of studies implicating the human microbiome in playing a critical role in many disease and wellness states. The etiology of abnormal semen analysis (SA) parameters is not identified in 30% of cases; investigations involving the semen microbiome may bridge this gap. Here, we explore the relationship between the semen microbiome and alterations of sperm parameters. We recruited men presenting for fertility evaluation or vasectomy consultation with proven biological paternity. SA and next generation sequencing was performed. Differential abundance testing using Analysis of composition of Microbiota with Bias Correction (ANCOM-BC) was performed along with canonical correlational analysis for microbial community profiling. Men with abnormal (N = 27) sperm motility showed a higher abundance of Lactobacillus iners compared to those with normal (N = 46) sperm motility (mean proportion 9.4% versus 2.6%, p = 0.046). This relationship persisted on canonical correlational analysis (r = 0.392, p = 0.011). Men with abnormal sperm concentration (N = 20) showed a higher abundance of Pseudomonas stutzeri (2.1% versus 1.0%, p = 0.024) and Pseudomonas fluorescens (0.9% versus 0.7%, p = 0.010), but a lower abundance of Pseudomonas putida (0.5% versus 0.8%, p = 0.020), compared to those with normal sperm concentration (N = 53). Major limitations are related to study design (cross-sectional, observational). Our results suggest that a small group of microorganisms may play a critical role in observed perturbations of SA parameters. Some of these microbes, most notably Lactobacillus iners, have been described extensively within other, fertility-related, contexts, whereas for others, this is the first report where they have potentially been implicated. Advances in our understanding of the semen microbiome may contribute to potentially new therapeutic avenues for correcting impairments in sperm parameters and improving male fertility.

Primordial germ cells (PGCs) are the embryonic precursors of sperm and eggs. They transmit genetic and epigenetic information across generations. Given the prominent role of germline defects in diseases such as infertility, detailed understanding of human PGC (hPGC) development has important implications in reproductive medicine and studying human evolution. Yet, hPGC specification remains an elusive process. Here, we report the induction of hPGC-like cells (hPGCLCs) in a bioengineered human pluripotent stem cell (hPSC) culture that mimics peri-implantation human development. In this culture, amniotic ectoderm-like cells (AMLCs), derived from hPSCs, induce hPGCLC specification from hPSCs through paracrine signaling downstream of ISL1. Our data further show functional roles of NODAL, WNT, and BMP signaling in hPGCLC induction. hPGCLCs are successfully derived from eight non-obstructive azoospermia (NOA) participant-derived hPSC lines using this biomimetic platform, demonstrating its promise for screening applications.

Despite rapid evolution across eutherian mammals, the X-linked MIR-506 family miRNAs are located in a region flanked by two highly conserved protein-coding genes (SLITRK2 and FMR1) on the X chromosome. Intriguingly, these miRNAs are predominantly expressed in the testis, suggesting a potential role in spermatogenesis and male fertility. Here, we report that the X-linked MIR-506 family miRNAs were derived from the MER91C DNA transposons. Selective inactivation of individual miRNAs or clusters caused no discernible defects, but simultaneous ablation of five clusters containing 19 members of the MIR-506 family led to reduced male fertility in mice. Despite normal sperm counts, motility, and morphology, the KO sperm were less competitive than wild-type sperm when subjected to a polyandrous mating scheme. Transcriptomic and bioinformatic analyses revealed that these X-linked MIR-506 family miRNAs, in addition to targeting a set of conserved genes, have more targets that are critical for spermatogenesis and embryonic development during evolution. Our data suggest that the MIR-506 family miRNAs function to enhance sperm competitiveness and reproductive fitness of the male by finetuning gene expression during spermatogenesis.

1. This study investigated the potential use of glycerol as a cryoprotectant for quail sperm cells. Its role in maintaining sperm fertilising ability in vivo and in vitro quality parameters, such as motility, was assessed. 2. The data showed that the presence of glycerol in semen samples was associated with infertility, which suggested that removal prior to insemination is mandatory. Removal through serial dilution centrifugation was associated with fewer than 5% of motile sperm cells and resulted in no fertility. 3. In conclusion, glycerol alone is not suitable for quail semen cryopreservation, and other approaches need to be investigated to develop cryobanking programmes for this species. [ABSTRACT FROM AUTHOR]

Purpose: Non-obstructive azoospermia (NOA) represents the persistent absence of sperm in ejaculate without obstruction, stemming from diverse disease processes. This survey explores global practices in NOA diagnosis, comparing them with guidelines and offering expert recommendations. Materials and Methods: A 56-item questionnaire survey on NOA diagnosis and management was conducted globally from July to September 2022. This paper focuses on part 1, evaluating NOA diagnosis. Data from 367 participants across 49 countries were analyzed descriptively, with a Delphi process used for expert recommendations. Results: Of 336 eligible responses, most participants were experienced attending physicians (70.93%). To diagnose azoospermia definitively, 81.7% requested two semen samples. Commonly ordered hormone tests included serum follicle-stimulating hormone (FSH) (97.0%), total testosterone (92.9%), and luteinizing hormone (86.9%). Genetic testing was requested by 66.6%, with karyotype analysis (86.2%) and Y chromosome microdeletions (88.3%) prevalent. Diagnostic testicular biopsy, distinguishing obstructive azoospermia (OA) from NOA, was not performed by 45.1%, while 34.6% did it selectively. Differentiation relied on physical examination (76.1%), serum hormone profiles (69.6%), and semen tests (68.1%). Expectations of finding sperm surgically were higher in men with normal FSH, larger testes, and a history of sperm in ejaculate. Conclusions: This expert survey, encompassing 367 participants from 49 countries, unveils congruence with recommended guidelines in NOA diagnosis. However, noteworthy disparities in practices suggest a need for evidence-based, international consensus guidelines to standardize NOA evaluation, addressing existing gaps in professional recommendations. [ABSTRACT FROM AUTHOR]

Hyaluronan (HA) is a glycosaminoglycan that forms a gel-like barrier in the subcutaneous (SC) space, limiting bulk fluid flow and the dispersion of SC-administered therapeutics. Recombinant human hyaluronidase PH20 (rHuPH20) facilitates the rapid delivery of co-administered therapeutics by depolymerizing HA in the SC space. Administration of rHuPH20 can induce the formation of anti-rHuPH20 antibodies, or anti-drug antibodies (ADAs), with the potential to bind endogenous PH20 hyaluronidase in the adult testes and epididymis. Using a variety of relevant animal models and multiple dose regimens of rHuPH20 across the full spectrum of animal development, we demonstrated that rHuPH20 administration resulted in the formation of ADAs. Although these ADAs can bind both the recombinant rHuPH20 enzyme and recombinant versions of animal model-specific hyaluronidases, they had no impact on fertility parameters (as measured by sperm concentration and motility, litter size, and litter viability) or fetal development. We present the result of our nonclinical studies in order of the developmental lifecycle, beginning with adults. Toxicology studies that extend beyond the standard package are also presented. These studies demonstrate the favorable safety profile of rHuPH20 and ADAs in nonclinical models. Additionally, we identified substantial safety margins for clinically relevant doses of rHuPH20. [Display omitted] • rHuPH20 facilitates subcutaneous administration of therapeutics. • Impact of rHuPH20 and anti-drug antibodies observed in multiple nonclinical models. • rHuPH20 anti-drug antibodies did not negatively alter fertility or development. • Substantial safety margins were identified when compared to human-equivalent doses. • Studies support favorable safety profile of rHuPH20. [ABSTRACT FROM AUTHOR]

Little is known about the effects of low gravity on human gametes. The aim of this study was to analyze if fresh human sperm samples after fast gravity load changes suffered any detrimental effect in comparison to the splits maintained in Earth's gravity. Fifteen fresh samples from normozoospermic donors were analyzed. Statistically significant differences in vitality (69.7 ± 9.9 % vs 72.4 ± 9.7 %, [95 % CI: 0.002,0.07]); motile sperm concentration (23.7 ± 15.3 M/ml vs 31.5 ± 25.1 M/ml, [95 % CI: 1.03,14.65]); grade "a" sperm concentration (8.7 ± 6.5 M/ml vs 11.7 ± 9.9 M/ml, [95 % CI: 0.71,5.28]); percentage of progressive motility sperm (30 ± 12.9 % vs 36 ± 14.3 %, [95 % CI: 0.10,0.37]) and curvilinear velocity VCL: 45.7 ± 12.8 μm/s vs 47.7 ± 13.3 μm/s, [95 % CI: 0.79,3.22]) were observed. No statistical differences were observed in other sperm kinematic parameters, morphology, DNA fragmentation, apoptosis, and oxidative stress. In conclusion, even though it did not result in a total loss, heavy gravity load changes including microgravity causes a significant decrease in sperm vitality and motility suggesting that negative consequences would be even higher if the exposure were longer. The results obtained indicate that further research is really needed before Assisted Reproduction will be considered for the future human reproduction outside the Earth. • Motility and Vitality of human sperm are significantly decreased after parabolic flight including short periods of microgravity. • Curvilinear Velocity of human male gametes was particularly decreased. • No changes were found in other sperm kinematic parameters, morphology, DNA fragmentation, apoptosis and oxidative stress. • Results were found after short exposure; suggesting more negative consequences regarding human fertility in a longer time exposure. • More studies needed before Assisted Reproduction Techniques can be considered for the likelihood of Human Reproduction in Space. [ABSTRACT FROM AUTHOR]

The decrease in fertility in aging roosters is related to the reduced quality of ejaculated sperm. This study aimed to investigate the effect of dietary supplementation with ginseng extract at various concentrations (0–150 mg/kg) on testicular function, semen preservation, and fertility at different stages of sexual maturity (mature and aging roosters) in Thai native roosters. Pradu Hang Dum roosters at 32 (mature; n = 24) and 75 (aging; n = 24) weeks of age were fed diets with non-supplemented or supplemented ginseng extracts (50, 100, and 150 mg/kg) until the end of the experiment. In experiment 1, fresh semen samples were examined for the quality parameters of semen volume, sperm concentration, sperm motility, sperm viability, lipid peroxidation, and enzymatic activities. In experiment 2, semen was preserved at 5 °C for up to 48 h, and the semen quality and fertility potential were determined. In experiment 3, testicular function and testosterone concentrations were evaluated. The results showed that ginseng extract supplementation in the diets of both mature and aging roosters at 50 and 100 mg/kg improved fresh semen quality (P < 0.05). A decrease in malondialdehyde levels in fresh semen was observed with increasing enzyme activities. In mature roosters, the progressive motility of cold-stored semen and fertility rates were higher in the G50 and G100 groups compared to the control and G150 groups after 24 h of storage (P < 0.05). In aging roosters, the highest significant differences in progressive motility, viability, and fertility rates were observed in the G50 and G100 groups at all storage times (P < 0.01). These improvements might be attributed to good testicular function in spermatogenesis, as revealed by the results of histological examination and testosterone concentrations. However, higher doses of ginseng extract supplementation negatively affected sperm quality. In summary, the recommended dose of ginseng extract supplementation in diets is 50 mg/kg. Fertility results indicated that insemination with semen preserved for 24 h was satisfactory in both mature and aging roosters. • Ginseng extract improved sperm production and semen preservation in mature and aging roosters. • Semen quality was highest in G50 and G100 groups, confirmed by histological and testosterone results. • High doses (150 mg/kg) negatively impacted sperm quality. • 24 hour preserved semen showed satisfactory fertility rates in both mature and aging roosters. [ABSTRACT FROM AUTHOR]

Introduction: Since boar spermatozoa show a marked deterioration in sperm quality when cooled, insemination doses are usually stored at 16-18 °C. However, maintaining this temperature during transport of semen doses is challenging, particularly during the summer months. An alternative could be to store the doses at 4 °C if cold-shock to the sperm could be prevented. The objective of this study was to evaluate boar sperm quality and fertility in in vitro fertilization after storage in AndroStar Premium at 4 °C for 1 week. Methods: Insemination doses (n =9) in AndroStar Premium from a commercial boar semen collection station were transported to the laboratory at approximately 20 °C. At the laboratory, sperm quality evaluation and was preformed and each dose was split; half of each ejaculate was stored in a climate-controlled box at 16-18 °C, the other was slowly cooled to 4 °C. Both samples were stored for 1 week before further sperm quality evaluation and in vitro fertilization (IVF) were performed. Mean values were tested using generalized linear regression, with treatment and boar as fixed factors; p ≤ 0.05 was considered significant. Results: Sperm membrane integrity (mean±sem: 91±0.05 and 83±0.09% for 16 and 4 °C, respectively) and superoxide production (6.79±2.37 and 13.54±6.23% for 16 and 4 °C, respectively), were different between treatments. The DNA fragmentation index was lower in cold-stored samples than in conventionally stored samples (3.74±2.25 and 7.40±3.36% for 4 and 16 °C, respectively). The numbers of oocytes developing to blastocyst on Day 6 (mean±sd: 9.0±8.0 and 6.0±5.0%, for storage at 16 and 4 °C, respectively) were not different between treatments. Discussion: Therefore, storage of boar semen doses in AndroStar Premium at 4 °C for up to 7days would be a viable alternative to current praxis. [ABSTRACT FROM AUTHOR]

Background: Valproic acid (VPA) is a widely used antiepileptic drug (AED) often prescribed as a first-line treatment for many idiopathic and symptomatic generalized epilepsies. Several studies have highlighted the side effects of VPA on male fertility and reproductive factors in males, although the specific underlying etiology of these abnormalities is not clear. The present systematic review and meta-analysis aimed to assess the preclinical and clinical evidence concerning the impact of VPA on male fertility and reproductive factors. Methods: The scientific literature was reviewed for eligibility using PubMed, Web of Science, and PsycINFO, encompassing preclinical and clinical studies. Factors related to male fertility and reproduction, such as differences in sperm count, sperm motility, and the percentage of abnormal sperm, were compared between the experimental groups treated with VPA (in both preclinical and clinical) and the control groups using the Standardized Mean Difference (SMD) with 95% confidence intervals (CIs). Additionally, differences in follicle-stimulating hormone (FSH) and luteinizing hormone (LH) were explicitly assessed in clinical studies. Results: Male fertility data were extracted from 7 preclinical studies (112 animals) and 5 clinical studies (274 male individuals). The results of animal studies found that the sperm count (SMD = -2.28, 95% CI: -3.39 to -1.18, P = 0.335) and sperm motility (SMD = -2.32, 95% CI: -3.34 to -1.30, P = 0.368) were decreased in the treated groups compared to the control groups. The percentage of abnormal sperm (SMD = 3.27, 95% CI: 1.98 to 4.56, P = 0.019) was significantly increased, while a non-significant reduction was revealed in the weight of the testis (SMD = -2.73, 95% CI: -4.23 to -1.23, P = 0.673) in treated groups. The outcomes of clinical studies indicated a non-significant decrease in sperm count (SMD = -0.78, 95% CI: -1.58 to 0.03, P = 0.286) among patients with epilepsy treated with VPA compared to control subjects. However, a significant reduction in sperm motility (SMD = -1.62, 95% CI: -2.81 to -0.43, P = 0.033 was observed. The percentage of abnormal sperm showed a non-significant increase (SMD = 0.93, 95% CI: -0.97 to 2.84, P = 0.616) after being treated with VPA. Furthermore, there was a non-significant reduction in the levels of FSH (SMD = -1.32, 95% CI: -2.93 to 0.29, P = 0.198) and LH (SMD = -0.96, 95% CI: -1.95 to 0.04, P = 0.211) observed in clinical participants. Conclusion: This meta-analysis of both preclinical and clinical studies revealed that VPA causes a significant reduction in male fertility and reproductive factors among male patients with epilepsy. Clinical neurologists should be more cautious when prescribing VPA, especially to young male adult patients with epilepsy. [ABSTRACT FROM AUTHOR]

Cooled semen storage methods result in oxidative stress generated by an imbalance between oxidation rates, specifically reactive oxygen species production, and sperm cell antioxidants, leading to degradation of semen quality. We aimed to investigate the impact of adding Eurycoma longifolia (EL) extract as an antioxidant supplement in semen storage medium (IGGKPh semen extender) on semen quality and fertility potential. EL extract at concentrations of 5, 10, 15, and 20 mg/mL was assessed for its antioxidant capacity in IGGKPh semen extender. Our findings revealed that the total phenolic content in the EL extract did not vary significantly across the various concentrations and temperatures tested. However, incubation at 5°C was found to be the most effective temperature for increasing the EL extract antioxidant capacity as assessed via the 2,2-diphenyl-1-picrylhydrazyl inhibition assay in a dosedependent manner. Supplementation of the IGGKPh semen extender with 15 mg/mL EL extract was found to enhance semen quality during cold storage for up to 48 h (p < 0.05), as evidenced by decreased malondialdehyde levels in cooled semen (p < 0.05). However, antioxidant enzyme activities showed no significant differences among the various experimental groups (p > 0.05). The fertility test showed that the 15 mg/mL EL extract group stored for 24 h had a higher percentage than the control group (p < 0.05). However, there was no significant difference in percentage between the two groups at 48 h of storage (p > 0.05). The hatchability showed no significant difference in both 24 and 48-h storage periods (p > 0.05). Our results indicated that supplementing the IGGKPh semen extender with 15 mg/mL EL extract may positively influence semen quality during storage, suggesting potential applications for enhancing semen quality. [ABSTRACT FROM AUTHOR]

Simple Summary: This study evaluated the impact of resveratrol-loaded cyclodextrin (RLC) on the quality of ram sperm after thawing. Our hypothesis was that resveratrol-loaded cyclodextrin, which enhances the antioxidant activity of resveratrol (RES), would improve the outcomes of freezing ram semen. Accordingly, this study investigates the effects of resveratrol at varying doses (10, 20, and 40 µg/mL), either in its pure form or loaded into methyl-β-cyclodextrin (CD), on sperm functions following the thawing of ram semen. The findings indicate that while cyclodextrin negatively affected sperm quality, particularly by increasing early capacitation and mitochondrial activity, resveratrol at a dose of 20 µg/mL effectively prevented early capacitation. Overall, both resveratrol and resveratrol-loaded cyclodextrin were generally effective and beneficial in the context of freezing ram semen. This study investigated the effects of pure and methyl-β-cyclodextrin loaded forms of resveratrol (10 µg/mL, 20 µg/mL, and 40 µg/mL) on ram sperm functions post-thawing. Semen samples were pooled and divided into ten groups: Control, RES10, RES20, RES40, CD10, CD20, CD40, RLC10, RLC20, and RLC40. The groups were pre-diluted with media containing the group-specific chemicals, followed by 15 min of incubation, dilution, and freezing. To assess the effects of the chemicals, a post-thaw sperm quality assessment was conducted. Motility and other velocity parameters were evaluated using computer-assisted semen analysis. The functional integrity of spermatozoa membranes was assessed with the hypo-osmotic swelling test, and the capacitation status of spermatozoa was determined through fluorescent microscopic evaluation. Additionally, flow cytometry was used to evaluate mitochondrial activity, oxidative stress, and the integrity of the sperm membrane and acrosome. The results indicated that cyclodextrin adversely affected sperm functions following freezing–thawing, notably increasing the rate of spermatozoa exhibiting pre-capacitation and mitochondrial activity by approximately 34% and 16%, respectively (p < 0.05). It was found that 20 µg/mL resveratrol prevented pre-capacitation (p < 0.05). Both resveratrol and resveratrol-loaded cyclodextrin groups improved post-thaw sperm qualities overall, demonstrating their utility for freezing ram semen. However, higher concentrations of resveratrol were found to negatively impact sperm functions. [ABSTRACT FROM AUTHOR]

Simple Summary: This study investigates the microbiome of bulls infected with Tritrichomonas foetus (T. foetus), a protozoan causing bovine tritrichomoniasis. It aims to describe the preputial, penile, and seminal microbiomes of chronically infected bulls and evaluate different sampling methods. Eleven bulls were sampled using dacron swabs, pizzle sticks, double-guarded swabs, and semen collection. The microbiomes were dominated by Fusobacteria, Firmicutes, Bacteroidota, Actinobacteria, and Campylobacterota, with semen samples showing the most microbial diversity. Despite differences in operational taxonomic units (OTUs) between methods, there was no significant difference in microbial richness and abundance. The study highlights that T. foetus infections may lead to shifts in the reproductive microbiome, indicating a potential dysbiosis compared to healthy bulls. These findings provide a basis for future research on T. foetus impacts on bovine reproductive health and possible treatment strategies. Tritrichomonas foetus (T. foetus), the causative agent of bovine trichomoniasis, is an obligate protozoan parasite of the bovine reproductive tract and can be found on the penis, prepuce, and distal urethra of the bull and from the cranial vagina to the oviduct in the infected cow. To date, the microbiome of bulls infected with T. foetus has not been described. The objectives of this study were to (1) describe the preputial and penile microbiome of bulls chronically infected by T. foetus, (2) describe the seminal microbiome of T. foetus-infected bulls, and (3) evaluate different collection devices that could be used for sampling. Eleven bulls naturally infected with T. foetus were utilized for the collection of samples. Samples were obtained during the process of a routine breeding soundness exam utilizing either a dacron swab, pizzle stick, double-guarded swab, or semen collection. The preputial and seminal microbiome of T. foetus-infected bulls was dominated by bacterial members of the phyla Fusobacteriota, Firmicutes, Bacteroidota, Actinobacteria, and Campylobacterota. Semen collection yielded the most microbial diversity; however, there was no significant difference between the four methods (p ≥ 0.05). This study characterizes both the preputial and seminal microbial communities of bulls chronically infected by T. foetus. [ABSTRACT FROM AUTHOR]

Sepsis is an inflammatory condition causing organ failure due to an uncontrolled immune response to infection and remains a significant challenge. Crotonis Semen has displayed various pharmacological effects, yet its potential in protecting against sepsis and the mechanisms involved remains largely unclear. Here, we explored the antiseptic properties of Crotons Semen extract (CSE) in both LPS-stimulated J774 macrophages and mice subjected to sepsis through Cecal ligation and Puncture (CLP) or LPS induction. We found that CSE enhanced survival rates in mouse models with acute sepsis induced by CLP operation and LPS injection. Administering CSE also reduced levels of enzymes indicating organ damage, such as aspartate aminotransferase (AST), alanine aminotransferase (ALT), and creatine kinase (CK), in septic mice. Furthermore, CSE lowered the serum levels of inflammatory mediators and cytokines, such as NO, TNF-α, IL-1β, and IL-6, in septic mice. In LPS-stimulated J774 macrophages, CSE reduced the expression of pro-inflammatory proteins, including iNOS and COX-2. Moreover, CSE inhibited the phosphorylation of IκBα and IKK, key components of the NF-κB signaling pathway, thereby reducing inflammatory mediators and cytokines. These results demonstrate CSE's protective effects against sepsis through NF-κB pathway disruption, indicating its potential as a therapeutic option for acute inflammatory conditions. [ABSTRACT FROM AUTHOR]

Background: Aromatase inhibitors improve male fertility by modifying the hormonal control of spermatogenesis. The present study aimed to investigate the effects of oral administration of letrozole on testosterone and estradiol concentrations and their ratios in blood serum, seminal plasma, prostatic fluid, sperm quality in fresh semen, and prostate gland dimensions. Seven adult male intact mixed-breed dogs were selected. The animals received letrozole (72 µg/kg, PO) daily for four weeks. Blood samplings and semen collections were carried out on days 0 (control), 14 (treatment), 28 (treatment), and 42 (post-treatment). Results: Our results showed that letrozole administration resulted in a 4.3 fold significant increase in serum, seminal plasma, and prostatic fluid testosterone levels after 14 days. This remained high until the end of the study. Serum and prostatic fluid estradiol levels did not change significantly over the study period. However, the seminal plasma estradiol level showed a significant increase on day 14. The estradiol: testosterone ratio was significantly reduced on day 14 in serum, seminal plasma, and prostatic fluid samples. Letrozole significantly improved the ejaculated spermatozoa viability and concentration after 28 days of oral administration. However, the sperm plasma membrane functional integrity and kinematic parameters were not significantly affected by the treatment. Transabdominal ultrasound examination revealed a significant increase in the height, width, and volume of the prostate gland after 28 days of treatment. Conclusions: According to the present research, oral administration of letrozole for 28 days affects local and systemic sex hormone balance leading to an improvement of the ejaculated canine spermatozoa viability and concentration concurrent with an increase in the prostate gland dimensions. [ABSTRACT FROM AUTHOR]

Context: Prostate-specific antigen (PSA) is important for semen liquefaction and sperm motility. Anti-PSA antibodies may lead to immune infertility. Aims: The present study was conducted to evaluate the impact of seminal anti-PSA antibodies on semen parameters in fertile and infertile men. Methods: A cross-sectional analytic study was conducted on 105 fertile men (≥21–50 years) having biological children (within last 2 years) with normal semen analysis as controls and 105 infertile men with abnormal semen analysis as cases. All semen samples were cryopreserved till 210 samples were collected, followed by estimation of anti-PSA antibodies using an indirect enzyme-linked immunosorbent assay technique. Key results: Mean ± standard deviation (s.d.) age of 210 participants was 30.0 ± 4.65 years. Mean ± s.d. levels of seminal anti-PSA antibodies in infertile men were 27.82 ± 102.19 ng/mL and in fertile men −30.45 ± 49.49 ng/mL (P = 0.001). A significant negative correlation was observed between anti-PSA antibody levels and sperm concentration (P = 0.013), rapid progressive motility (P = 0.001), slow progressive motility (P = 0.006), progressive sperm motility (P = 0.001), and normal morphology (P = 0.001), and significant positive correlation was observed with immotile sperms (P = 0.001). The overall accuracy of anti-PSA antibody for differentiating infertile from fertile men was 63.33%. Conclusions: Seminal anti-PSA antibodies were significantly correlated with semen parameters in fertile and infertile men with an accuracy of 63.33%. A negative correlation was observed between antibody levels and progressive sperm motility. Implications: Seminal anti-PSA antibodies can be used as a biomarker for male infertility assessment. Globally, male factor infertility is increasing and so is the need for newer and effective tests for its diagnosis and management. The present study was conducted to evaluate the impact of seminal anti-prostate-specific antibodies (PSA) antibodies on semen parameters in fertile and infertile men. A significant negative correlation was observed with sperm concentration, motility, and morphology, and a positive correlation was observed with grade D motility. The overall accuracy of anti-PSA antibodies in predicting infertile from fertile men was 63.33%. Image by Naina Kumar. [ABSTRACT FROM AUTHOR]

The objective of this study was to compare the plasma (PL) and seminal plasma (SP) pharmacokinetic profile of ceftiofur (CEFT) and desuroylceftiofur acetamide (DFCA) after administration of CEFT crystalline-free acid (CCFA) by SC route in two sites of the ear in beef bulls. Four clinically healthy Hereford bulls received a comprehensive physical exam and subsequently a breeding-soundness examination, CBC, and chemistry profile panel. All bulls were diagnosed healthy and satisfactory potential breeders. In one group (n = 2), a single dose of CCFA was administered SC route at the base of the ear (BOE) at a dose of 6.6 mg/kg of body weight. The second group (n = 2) was also administered by SC route in the middle third of the posterior aspect of the ear (MTE). The concentrations of CEFT and DFCA in PL and SP were determined by a high-performance liquid chromatography mass spectrometry (HPLC-MS). Blood and semen samples were collected before the administration of CCFA and at 12, 24, 36, 48, 72, 96, 120, 144, and 168 h after injection. No levels of CEFT were detected in PL and only in 20 of the 40 SP samples (P = 0.0001). The mean level of CEFT in SP was 0.11 % in comparison with the DFCA level. DFCA was found in all PL and SP samples. Therefore, DFCA was chosen to be utilized in the study of the pharmacokinetics parameters both in PL and SP. There were no differences in the mean PL levels of DFCA for the two sites of SC administration between the BOE (102.9 ± 78.9 ng/mL; X ± SD) and to MTE (116.1 ± 70.2 ng/mL; P = 0.58). The mean SP levels of DFCA after administration in the BOE was 857 ± 747 ng/mL, and for the MTE was 549 ± 488 ng/mL without differences between both sites (P = 0.15). The mean level of DFCA in PL was 109.5 ± 74.0 ng/mL, which was lower than the mean SP levels of 695 ± 103 ng/mL (P = 0.001). Moreover, the PL peak DFCA concentration (C max) was 229 ± 46 ng/mL at 36.0 ± 29.4 h (T max) post-administration. The SP C max was 1851 ± 533 ng/mL at 30.0 ± 28.6 h (T max) post-administration. The C max between PL and SP were distinctive (P = 0.004) without any differences in T max between PL and SP (P = 0.60). The terminal half-life for PL DFCA (47.4 ± 29.3 h) was not different than in SP (53.1 ± 23.6 h; P = 0.77). The PL area under the curve concentration time from the first to the last sample (AUC 0–last) was 18,984 ± 4841 ng/mL/h, which was significatively smaller compared with 125,677 ± 59,445 ng/mL/h for SP AUC 0–last (P = 0.04). The PL mean residence time from the first to the last sample (MRT 0–last) was 69.7 ± 15.1 h, and it was similar than for SP of 66.5 ± 7.7 h (P = 0.69). From the present investigation, based in its pharmacokinetic features, it was concluded that CCFA should be an appropriate antibiotic that could be used for the treatment of bull genital infections when its indication is properly outlined. To study the pharmacokinetics of CCFA in SP, DFCA metabolite was appropriated. [ABSTRACT FROM AUTHOR]

Breeding soundness evaluation (BSE) is the best methodology to estimate the fertility potential of future bulls and performing indirect selection for their fertility. However, the outcome of the BSE is influenced by several factors, including genetics, environment, and BSE guidelines. Herein, in this retrospective study, our first aim was to characterize the reasons for failure in 46,566 BSE from 2-year-old beef Bos indicus bulls (Nellore) born from 1997 to 2018. Our second aim was to determine whether or not BSE was associated with reproductive potential improvement of the bulls over the years. Due to changes in the BSE criteria, we used the same dataset, but only bulls born from 2002 to 2018 were included resulting in 35,856 BSE. For the second aim, the effect of the year and farm were included in the model of the multivariate logistic regression. We also determined if the main reasons for BSE failure decreased over time. Bulls were classified as approved (satisfactory potential breeders and qualified for natural breeding service) and not approved (deferred and unsatisfactory potential breeders). The reasons for BSE failure in Nellore bulls were poor semen quality (53.1 %) and physical defects (46.9 %), with the main physical defect being testis abnormalities (19.7 %). The overall percentage of bulls approved each year was 87.1 %, with no improvement over the years of study. However, the percentage of approved bulls at the first BSE increased over the years (P < 0.05). This increase was evident by a reduction in the difference between the overall percentage of the bulls approved vs the percentage of bulls approved at the first BSE. Furthermore, there was an increase in the percentage of bulls classified as satisfactory potential breeders in the BSE and an evident decrease in the percentage of bulls qualified only for natural breeding service (P < 0.05). In addition, an increase of the scrotal circumference (SC) of the herd was found (P < 0.05). These results indicate the overall quality of the bulls improved over the years. To associate and identify the main sperm abnormalities, 3461 not approved bulls were clustered. The most frequent defects were strongly coiled tail spermatozoa, proximal droplets, and acrosomal defects. Overall, there was no change in the frequency of bulls not approved by the sperm morphology nor the frequency of the main sperm abnormalities over the years. Nevertheless, the frequency of the defects remained very low, implying they were controlled. Additionally, abnormalities in the testis decreased over the years and was associated with the increase in the SC of the herd and a decrease of culled bulls due to low SC. In conclusion, this study demonstrates that there is an association between implementation and use of BSE with improvements in the reproductive quality of future generation bulls. • Records of 20 years and 46,566 BSE from a seedstock herd in Brazil were investigated. • About 14 % of the bulls fail to be approved in the BSE at two years of age. • Semen quality (53 %) & physical defects (47 %) were the most common causes of failure. • Testicular abnormalities decreased, & scrotal circumference increased over the years. • BSE was associated with increased percentage of bulls approved at first BSE over years. [ABSTRACT FROM AUTHOR]

Cryopreservation is a pivotal technique in safeguarding genetic material across diverse species, despite its inherent challenges linked to induced spermatozoa damage, notably apoptosis and lipid peroxidation (LPO). Given the insufficient antioxidant defense of spermatozoa against LPO, there is a rising interest in integrating additional additives into extenders to ameliorate mammalian semen quality. Among these additives, flavonoids have garnered considerable attention due to their potent antioxidative properties. Hence, our study aimed to assess the efficacy of flavone (FL) and 3-hydroxyflavone (3-OH =) supplementation in the cryopreservation medium to protect canine sperm against the damaging impacts of freezing and ensure the preservation of their reproductive potential. Semen was collected from five Beagle stud dogs and then pooled. Then, the sample was divided into 7 groups, each treated with 1) 0 mM, 2) 0.1 mM FL, 3) 0.2 mM FL, 4) 0.4 mM FL, 5) 0.1 mM 3-OH = , 6) 0.2 mM 3-OH = , 7) 0.4 mM 3-OH =. Semen samples were subjected to cryopreservation in French straws and glycerol as a cryoprotectant. In the frozen thawed semen, sperm motility parameters by CASA system and sperm membrane integrity, acrosome status, mitochondrial activity, DNA fragmentation, early apoptosis with capacitation, and LPO were assessed using flow cytometry just after thawing (0 h) and 4 h post thaw. Results reveal significant increase in the proportion of live spermatozoa with undamaged acrosomes in the FL 0.1 and 3-OH = 0.2 groups at 0 h post thaw. At this time point, 3-OH = 0.1 significantly reduced the DNA fragmentation index (DFI) compared to the FL 0.1 and 0.2 groups. However, after the next 4 h, 3-OH = 0.4 exhibited the lowest (P < 0.05) DFI compared to FL 0.2 and 3-OH = 0.1. Additionally, 3-OH = 0.4 showed the highest (P < 0.05) proportion of non apoptotic and non capacitated spermatozoa compared to FL 0.1 0 h post-thaw. Simultaneously, the same group demonstrated significant reduction in apoptotic and capacitated sperm cells, at 0 h and 4 h post-thaw. Moreover, 3-OH = at 0.1 (0 h and 4 h) and 0.2 mM (4 h) significantly enhances the proportion of live sperm without LPO post thaw. Whitin the FL groups, only 0.4 FL significantly increased the percentage of live sperm without LPO. No significant effect of the tested substances was observed on sperm motility, cell membrane integrity, or mitochondrial activity. These findings highlight the promising role of flavone and 3-hydroxyflavone in enhancing sperm resilience during cryopreservation, suggesting their protective function against acrosome damages, capacitation, apoptosis and lipid peroxidation. [Display omitted] • lFlavone (FL) and 3-hydroxyflavone (3-OH =) were used to supplement the extender for canine semen cryopreservation for the first time. • Using FL 0.1 mM and 3-OH = 0.2 mM increased (P < 0.05) the proportion of live spermatozoa with undamaged acrosomes. • Using 3-OH = 0.1 and 0.4 mM significantly reduced the DNA fragmentation index of spermatozoa. • Using 3-OH = 0.4 mM significantly reduced apoptosis and capacitation of sperm cells. • Using FL 0.4 mM and 3-OH = 0.1 and 0.2 mM significantly protected the sperm against lipid peroxidation. [ABSTRACT FROM AUTHOR]

Single Layer Centrifugation (SLC) through a low density colloid offers an alternative solution to antibiotic use in boar semen extenders, with lower costs compared to high density colloids. The aim of this study was to explore the reproductive performance of sows when using SLC-prepared semen doses without antibiotics, employing low density Porcicoll to prepare semen doses for artificial insemination in a commercial swine herd in Thailand. Ejaculates were divided into two equal parts to create insemination doses, with each dose containing 3000 × 106 sperm/80 ml for intra-uterine insemination in individual sows. The sows were inseminated twice, with the interval between the two inseminations ranging from 8 to 16 h. The CONTROL group consisted of 206 semen doses treated with antibiotics, prepared for insemination in 103 sows, while the SLC group comprised 194 SLC-prepared semen doses without antibiotics for inseminating 97 sows. Fertility and fecundity traits, including non-return rate, conception rate, farrowing rate, and litter traits (i.e., the total number of piglets born per litter, number of piglets born alive per litter, number of stillborn piglets, and number of mummified fetuses), were compared between groups. Furthermore, data on piglet characteristics, including live-born and stillborn piglets (i.e., the prevalence of stillbirth (yes, no), birth weight, crown-rump length, body mass index (BMI), and ponderal index (PI)), were determined. No significant differences in non-return rate (75.7 % vs. 77.3 %), conception rate (73.8 % vs. 73.2 %), and farrowing rate (71.8 % vs. 73.2 %) were observed between the CONTROL and SLC groups, respectively (P > 0.05). Nevertheless, the total number of piglets born per litter in the SLC group was higher than in the CONTROL group (14.6 ± 0.9 vs. 12.3 ± 0.6, respectively, P = 0.049). Interestingly, the prevalence of stillbirth in the SLC group was lower than in the CONTROL group (6.2 % vs. 11.6 %, respectively, P < 0.001). Moreover, the newborn piglets in the SLC group exhibited higher birth weight and BMI compared to those in the CONTROL group (1.36 ± 0.03 vs. 1.26 ± 0.02 kg, P = 0.005, and 18.3 ± 0.3 vs. 17.3 ± 0.2 kg/m2, P = 0.003). In conclusion, employing sperm doses after SLC through a low density colloid in artificial insemination within a commercial breeding operation did not have a detrimental impact on either fertility or fecundity traits but showed potential benefits in increasing the total number of piglets born per litter. Moreover, improvements were observed in the birth weight and body indexes of piglets, and the percentage of stillbirths was reduced. Our findings introduce new possibilities for antibiotic alternatives in semen extenders to reduce the risk of antimicrobial resistance in the swine industry. Additionally, they provide compelling reproductive outcomes supporting the integration of SLC-prepared semen doses into artificial insemination practices. • Single Layer Centrifugation is an alternative solution to antibiotic use in boar semen extenders. • Farrowing rate did not differ between control and SLC groups. • The total born in the SLC group was higher than in the control group. • The prevalence of stillbirth in the SLC group was lower than in the control group. • Newborn piglets in the SLC group had higher birth weight and BMI than control group. [ABSTRACT FROM AUTHOR]

Melatonin, a hormone synthesized in various tissues, plays a crucial role in modulating sperm characteristics, yet its protective function on boar sperm remains poorly understood. This study aimed to investigate the expression and localization of melatonin-related proteins (AANAT, ASMT, MT1, MT2, and NQO2) in pig tissues, assess the impact of melatonin on pig sperm motility parameters and quality, and elucidate the underlying molecular mechanisms. Our results revealed widespread expression of AANAT, ASMT, MT1, MT2, and NQO2 proteins in pig tissues, particularly in the testis. Specific localization patterns were observed in Leydig cells, reproductive epithelium, and columnar epithelium cells in the testis and cauda epididymis. Additionally, melatonin membrane receptors MT1 and MT2 were detected in boar sperm. Melatonin treatment significantly enhanced boar sperm motility parameters and quality, particularly with 10 nM melatonin treatment. Inhibition of the MT1 receptor, but not the MT2 receptor, resulted in decreased sperm motility, highlighting the pivotal role of the MT1 receptor in mediating melatonin's effects on boar sperm. Metabolomic analysis revealed significant alterations in sperm metabolites following melatonin supplementation, particularly in amino acid metabolism. Overall, our findings provide comprehensive insights into melatonin's mechanisms in improving boar sperm quality, suggesting its potential as a therapeutic agent for enhancing male fertility. • Comprehensive study shows melatonin boosts boar sperm motility and quality, mainly via the MT1 receptor. • Melatonin-related proteins in pig tissues suggest roles in testicular physiology and sperm protection. • Melatonin alters seminal plasma metabolites, showing diverse impacts on sperm function and metabolism. [ABSTRACT FROM AUTHOR]

Background: Centrifugation is a common procedure to improve the quality of chilled and frozen canine semen by removing debris and seminal plasma and adding semen extenders. The aim of this study was to evaluate the efficacy and influence of a second centrifugation after 48 h of storage at 5 °C on the sperm quality of canine semen. The ejaculates of 45 healthy male dogs, divided into three groups according to body weight, were analyzed for macro- and microparameters such as ejaculate volume, sperm concentration, kinematic parameters, morphology, and integrity of plasma membrane. Samples were analyzed at baseline conditions (T0), after 24 h (T24) and after 48 h (T48) to assess the effects of the different treatments on sperm quality. Results: The results showed a significant effect of a second centrifugation on the improvement of chilled sperm quality compared to the other techniques, especially up to 48 h. Conclusions: Analysis of the data showed that the semen samples centrifuged and then cooled at 5 °C had acceptable semen parameters, especially in terms of motility, with a gradual decrease in serial evaluations after 24 and 48 h. A second centrifugation after 48 h of storage may lead to better semen quality and improve the kinetics of sperm parameters, the percentage of morphologically normal sperm and the percentage of sperm with intact membranes. [ABSTRACT FROM AUTHOR]

Myo‐inositol may be efficient to improve sperm parameters to increase the chance of fertility. Although, the data are controversial. This study aimed to assess the impact of Myo‐inositol supplements on semen quality and in vitro fertilization (IVF) outcomes. In this systematic review and meta‐analysis, a comprehensive search was conducted in PubMed, Web of Science, and Embase. The objective was to identify relevant human studies that investigated the effects of Myo‐inositol treatment on various sperm factors, such as sperm motility, sperm concentration, sperm morphology, viable spermatozoa, spermatozoa with DNA fragmentation, and pregnancy rate. Additionally, the testosterone levels of patients with Oligo‐astheno‐teratozoospermia (OAT) after Myo‐inositol application were considered. The findings of 16 selected studies from 2240 citations indicated significant improvements in several parameters of sperm after Myo‐inositol administration. Myo‐inositol treatment was associated with a notable increase in total sperm motility (SMD 0.90; 95% CI: 0.34 to 1.46; I2 = 0%, p = .001) and progressive sperm motility (SMD 1.48; 95% CI: 0.37 to 2.59; I2  = 0%, p = .008). Additionally, there was a significant improvement in testosterone levels (SMD 0.54; 95% CI: 0.34 to 0.73; I2  = 0%, p < .0001). Furthermore, Myo‐inositol therapy demonstrated a significant decrease in spermatozoa with DNA fragmentation (SMD −1.37; 95% CI: −2.43 to −0.32; I2  = 85%, p = .01). This study suggests that Myo‐inositol therapy has a positive impact on specific sperm parameters, such as total and progressive sperm motility, along with testosterone levels. These findings provide support for the potential benefits of Myo‐inositol in improving male fertility parameters related to sperm factors. [ABSTRACT FROM AUTHOR]

Simple Summary: When a couple faces infertility challenges like sperm motility issues, they often visit fertility clinics for assistance. Poor sperm motility can delay fertilization, making natural conception difficult. In an andrology laboratory, embryologists test the semen sample. Before assisted reproduction procedures, embryologists usually perform a group of steps to prepare the semen sample and to isolate the best sperms. These technical steps might harm the sperms, because they become susceptible to something so called; oxidative stress, which occurs when too many harmful molecules called free radicals accumulate in the test tube, leading to cell damage and death. A group of chemicals called antioxidants can neutralize these free radicals. The researchers in this study investigated the therapeutic effects of Mitoquinone, which is a mitochondria-targeted antioxidant, on the motility of human sperms exposed to oxidative stress during in vitro procedures. Mitoquinone as an antioxidant might have therapeutic potential for sperms' oxidative-damage-associated disorders. This investigation paves the route for future research to confirm the beneficial role of Mitoquinone supplementation in improving and enhancing sperm quality during in vitro processes in the andrology laboratories. The preparation of human sperm in an andrology laboratory subjects it to oxidative stress. Reactive oxygen species are produced by mitochondria, making it susceptible to oxidative damage; hence, mitochondria-targeted antioxidants like Mitoquinone (MitoQ) might have therapeutic potential for oxidative-damage-associated disorders. The current research aims to establish whether MitoQ has any positive effects during in vitro preparation of fresh human sperm. Viability and motility are evaluated to determine the effective MitoQ concentration and to assess whether MitoQ supplementation is affected by sperm concentration by incubating normospermia semen samples at 37 °C for 2 h and 4 h, respectively. The effect of semen centrifugation following supplementation of 20 × 106 sperm/mL with 200 nM MitoQ is also assessed by measuring viability, motility and sperm DNA fragmentation. The best sperm motility is achieved after 2 h of incubation with 200 nM MitoQ at 37 °C. Sperm concentration of 20 × 106 sperm/mL is the best concentration where 200 nM MitoQ works efficiently. For semen centrifugation at 300× g for 20 min, supplementation with 200 nM MitoQ shows higher sperm motility. The current results demonstrate that MitoQ supplementation during in vitro human semen preparation procedures positively affects fresh sperm motility without affecting viability or increasing DNA fragmentation. [ABSTRACT FROM AUTHOR]

Simple Summary: In the Yucatán Peninsula, the availability and diversity of plants as food sources for bees vary throughout the year, which impacts dietary composition during the larval phase and compromises the development of their reproductive organs. We evaluated the eversion of the copulatory apparatus, ejaculation, and sperm quality of drones bred during flowering-scarcity (November–December) and flowering-onset periods (January–February). It was observed that more drones had a complete eversion of the copulatory apparatus during the flowering-onset period, with better ejaculation and sperm quality than those bred during the period of flowering scarcity. Insufficient protein intake during the larval phase of drones affects the development of reproductive organs and spermatogenesis. The aim of the study was to evaluate the effect of the flowering-scarcity and flowering-onset periods on the eversion of the copulatory apparatus, ejaculation, and sperm quality of drones (Apis mellifera L.). To stimulate the laying of drone eggs during the flowering-scarcity period, 1 L of sugar syrup was supplied weekly to the hives, along with a protein supplement made of 20% pollen and 80% brewer's yeast. During the periods of flowering scarcity and the onset of flowering, 800 drones were collected (200 per month). At the onset of flowering, 270 drones showed eversion of the copulatory apparatus, of which 162 ejaculated, compared to the flowering-scarcity period, where 112 drones showed eversion of the copulatory apparatus and 39 drones ejaculated. During the period of flowering onset, sperm parameters such as volume, concentration, motility, viability, acrosome integrity, and mitochondrial activity were higher compared to the period of flowering scarcity. In conclusion, we observed a greater number of drones with eversion of the copulatory apparatus and ejaculation, as well as better sperm quality with the drones bred during the period of flowering onset. These differences indicate that the flowering-scarcity period significantly impacts the reproductive parameters of drones. [ABSTRACT FROM AUTHOR]

The objective of this study was to determine the effect of a coconut water-based extender on the quality parameters of dog semen (n=24 ejaculates) during refrigeration (4°C) preservation up to 72 h. Four ejaculates per dog were collected from 6 dogs by digital manipulation. All fresh ejaculates were evaluated for macroscopic and microscopic semen evaluation tests. The semen samples were then extended in a coconut water-based extender using a split sample technique. The diluted semen was kept in a refrigerator for preservation at 4°C and evaluated at 0, 24, 48, and 72 h for sperm quality parameters. The sperm motility significantly reduced over a period of time from 0 to 72 h. The live sperm percentage did not differ significantly at 0, 24 and 48 h of preservation. However, there was a significant drop in live sperm percentage at 72 h. The percent sperm abnormalities were significantly increased from 24, 48 to 72 h of preservation. In coconut water-based extender there was a significant difference in intact plasma membrane percentage at different time slots (0, 24, 48 and 72 h). It was concluded that coconut water-based extender can preserve canine sperm for up to 48 h while maintaining sperm motility and viability above 50%. Sperm abnormalities remained within 20% and plasma membrane integrity above 60% both were maintained up to 72 h of semen preservation. [ABSTRACT FROM AUTHOR]

The addition of antioxidants to cryopreservation media reportedly improves sperm post‐thaw quality and reproductive performance after artificial insemination. Therefore, the objectives of this study were to evaluate if the addition of L‐carnitine and pyruvate to freezing media, or their addition to samples after thawing, improves the post‐thaw quality of equine spermatozoa. Thus, in Experiment 1, stallion semen samples were cryopreserved in: (1) EDTA–glucose–based extender with 20% egg yolk and 5% dimethylformamide (EDTA control); (2) skim milk–based extender with 20% egg yolk and 5% dimethylformamide (milk control); (3) Extender 1 supplemented with 50 mM L‐carnitine and 10 mM pyruvate (EDTA‐carnitine‐pyruvate); and (4) Extender 2 supplemented with 50 mM L‐carnitine and 10 mM pyruvate (milk‐carnitine‐pyruvate). In Experiment 2, 50 mM L‐carnitine and 10 mM pyruvate were added post‐thaw to samples cryopreserved with extenders 1 and 2 (EDTA control and milk control). Sperm kinematic parameters, DNA fragmentation, membrane lipid peroxidation, acrosome status and viability were evaluated after thawing. No significant differences (p > 0.05) were observed for most of the kinematic parameters, DNA fragmentation, membrane lipid peroxidation, acrosome status and viability of spermatozoa, between the samples frozen in the presence or absence of L‐carnitine and pyruvate, nor between the samples after the post‐thaw addition of these components. A higher (p < 0.05) mean velocity and higher (p < 0.05) amplitude of lateral head displacement were observed in the samples frozen in the milk‐based extender with the addition of L‐carnitine and pyruvate after thawing. The addition of 50 mM L‐carnitine and 10 mM pyruvate, either to the freezing extenders or after thawing, was not deleterious for sperm; however, it did not improve equine sperm motility, viability, acrosome and DNA integrity, nor decrease membrane lipid peroxidation after thawing. [ABSTRACT FROM AUTHOR]

Trichomonas vaginalis and associated bacteria are important urogenital pathogens. This study aimed to investigate the effect of T. vaginalis and its associated bacteria on semen and sperm parameters related to male fertility. Urine and semen samples were collected from 97 males, whose ages between 18-50 years. Semen and sperm parameters, such as semen volume, sperm count, sperm concentration, sperm motility, leucocytes/pus cells, and erythrocytes, were evaluated according to the WHO instructions. Conventional-PCR and real-time quantitative PCR were used to detect T. vaginalis infection. Amie's-transport, blood-agar, MacConkey-agar, Chocolate-agar, Thayer-Martin-agar, and Mannitol-salt-agar were used to culture bacteria in urine, and the Vitek2 system was used to differentiate bacteria. Samples that showed positive microscopic examination for bacteria but were not culturable were diagnosed using PCR. T. vaginalis and bacterial infection rates were 31.8% and 17.5%, respectively. T. vaginalis, bacteria, and both together caused a decrease in some qualitative and quantitative parameters of semen and sperm such as a decline in sperm total count by 46.5, 63.46, and 65.67% respectively, sperm concentration by 6.27, 15.37, and 44.27% respectively, sperm motility by 67.63, 66.18 and 66.75% respectively, semen volume by 42.2, 36.4 and 63.86% respectively. They caused increases of 100.92, 16.89, and 84.56%, respectively, and sperm death by 36.58, 80.0%, 93%, and 60.77%, respectively. In addition, they caused an increase in WBCs/pus cells and RBCs in the semen and urine. Four bacteria species, Pantoea spp., Sphingomonas paucimobilis, Staphylococcus aureus, Methylobacterium spp. were detected in urine, and 10 species, Enterobacter aerogenes, Serratia marcescens, Pseudomonas putida, Sphingomonas paucimobilis, Aeromonas sobria, Burkholderia cepacia, Staphylococcus aureus, Aerococcus viridans, Pseudomonas androgens and Staphylococcus epidermidis were detected in semen. In addition, PCR identified three unculturable species: one in urine (Holomonas spp.), and two in semen (Lactobacillus jensenii and Holomonas spp.), S. aureus had the highest frequency in urine and semen of (32%). [ABSTRACT FROM AUTHOR]

Sperm DNA fragmentation (sDF) is a DNA damage able to predict natural conception. Thus, many laboratories added tests for the detection of sDF as an adjunct to routine semen analysis with specific indications. However, some points related to sDF are still open. The available tests are very different each from other, and a direct comparison, in terms of the prediction of reproductive outcomes, is mandatory. The proposed mechanisms responsible for sDF generation have not yielded treatments for men with high levels of sDF that have gained the general consent in clinical practice, thus requiring further research. Another relevant point is the biological meaning to attribute to sDF and, thus, what we can expect from tests detecting sDF for the diagnosis of male infertility. SDF can represent the "tip of iceberg" of a more extended and undetected sperm abnormality somehow impacting upon reproduction. Investigating the nature of such a sperm abnormality might provide novel insights into the link between sDF and reproduction. Finally, several studies reported an impact of native sDF on assisted reproduction technique outcomes. However, to fertilise the oocyte, selected spermatozoa are used where sDF, if present, associates with highly motile spermatozoa, which is the opposite situation to native semen, where most sDF associates with non-viable spermatozoa. Studies comparing the impact of sDF, as assessed in both native and selected spermatozoa, are needed. [ABSTRACT FROM AUTHOR]

Simple Summary: This study compares two methods of preparing boar sperm for in vitro fertilization: the traditional density gradient selection (DGS) using centrifugation and the newer microfluidic chip-based sperm (MCS) sorting. MCS resulted in lower sperm concentration and fewer morphologically abnormal sperm compared to DGS. Additionally, although DGS showed higher progressive motility, both methods had similar fertilization outcomes in terms of cleavage rates, blastulation rates, and embryo quality. The results show that MCS performs at least as well as DGS and may offer advantages due to its gentler approach and potential for more consistent results in IVF procedures. Porcine in vitro embryo production (IVP) protocols have conventionally used density gradient selection (DGS) by centrifugation to prepare sperm samples and achieve successful fertilisation. However, the possible toxicity of the solutions used and the potential damage caused by the centrifugation step may have a negative effect on the quality of the sample. Microfluidic chip-based sperm (MCS) sorting has been proposed as an alternative technique for the selection of high-quality sperm with the purpose of improving reproductive outcomes in IVF. This device does not require centrifugation or any toxic solution to prepare the sample for fertilisation. The sample is not subjected to unnecessary stress, and the process is less operator-dependent. In this study, we compared the sperm parameters of unselected extender-diluted boar semen samples with selected samples using DGS and MCS methods. The results show an expected reduction in sperm concentration after both methods. All the groups were significantly different from one another, with MCS being the group with the lowest concentration. Though the three groups had a similar overall motility, significant differences were found in progressive motility when comparing the unselected group (control, 19.5 ± 1.4%) with DGS and MCS. Progressive motility in DGS was also significantly higher than in MCS (65.2 ± 4.9% and 45.7% ± 5.3, respectively). However, MCS selection resulted in enriched sperm samples with a significantly lower proportion of morphologically abnormal sperm compared to DGS. After fertilisation, no statistical differences were found between the two methods for embryological parameters such as cleavage rates, blastulation rates, and embryo quality. The number of cells in blastocysts derived from MCS was significantly greater than those derived from DGS sperm. Thus, we demonstrate that MCS is at least as good as the standard DGS for most measures. As a more gentle and reproducible approach for sperm selection, however, it could improve consistency and improve IVP outcomes as mediated by a greater proportion of morphologically normal sperm and manifested by a higher cell count in blastocysts. [ABSTRACT FROM AUTHOR]

Simple Summary: The timing of ovulation varies widely among gilts, leading farms to often perform multiple inseminations to ensure that sperm and eggs meet in the reproductive tract, thus enhancing fertility. Administering a gonadotropin-releasing hormone (GnRH) agonist, such as buserelin, at the onset of estrus can induce ovulation and reduce the variation in ovulation timing among sows. This study examined the effect of adding buserelin to boar semen on the litter size of gilts. Gilts were inseminated with semen enriched with either 5 or 10 µg of buserelin acetate during the initial insemination. Although there were no significant differences in the farrowing rate between the groups, the group receiving 10 µg of buserelin acetate had a higher total number of piglets and live-born piglets. Additionally, this group exhibited greater litter birth weight compared to the control group. These findings indicate that supplementing boar semen with a GnRH agonist enhances litter size in gilts. The use of exogenous hormones has long been of interest for improving reproductive performance in swine production. Enhancing litter size directly impacts the economic efficiency of pig production. Various strategies, including nutritional, genetic, and hormonal approaches, have been explored with varying degrees of success. Administering a gonadotropin-releasing hormone (GnRH) agonist, such as buserelin, at the onset of estrus can induce ovulation and reduce the variation in ovulation timing among sows. This study assessed the impact of GnRH agonist supplementation in boar semen doses on the litter size of inseminated gilts. The research was conducted on a commercial swine herd in northern Thailand. A total of 231 Landrace × Yorkshire crossbred gilts, aged 224.5 ± 16.2 days at the onset of estrus synchronization, participated in the experiment. The gilts' estrus was synchronized with oral altrenogest supplementation at a dosage of 20 mg/day for 18 days. After exhibiting standing estrus, the gilts were randomly divided into three groups. Control group: gilts were inseminated at 0 and 12 h post standing estrus onset with a conventional semen dose (n = 94). Treatment 1: similar to the control group, but with an added 5 µg (1.25 mL) of buserelin acetate to the boar semen dose during the first insemination (n = 71). Treatment 2: similar to the control group, but with 10 µg (2.5 mL) of buserelin acetate added to the boar semen dose during the first insemination (n = 66). All gilts were inseminated twice during their standing estrus using the intrauterine artificial insemination method. Each semen dose contained 3.0 × 109 motile sperm in 80 mL. The farrowing rate averaged 78.8% and did not significantly differ between the groups (p = 0.141). The total number of piglets born per litter in the treatment 2 group was greater than in the control group (14.0 ± 0.3 vs. 13.2 ± 0.3, respectively, p = 0.049), but was not significantly different from the treatment 1 group (13.3 ± 0.3, p = 0.154). Similarly, the number of live-born piglets in the treatment 2 group was greater than in the control and treatment 1 groups (13.2 ± 0.4 vs. 12.3 ± 0.3 and 12.0 ± 0.4, respectively, p < 0.05). Moreover, the live-born piglets' litter birth weight in the treatment 2 group was greater than in the control group (17.0 ± 0.4 vs. 15.6 ± 0.3 kg, respectively, p = 0.008) and the treatment 1 group (15.7 ± 0.4 kg, p = 0.025). In conclusion, adding a GnRH agonist to boar semen appears to enhance the litter size of gilts. Further research should focus on understanding the underlying mechanisms and determining the optimal dose and timing for GnRH agonist supplementation. [ABSTRACT FROM AUTHOR]

Context: The conflicting findings regarding the impact of equilibration time on post-thawed sperm quality underscore the need for further research to evaluate the impact of equilibration time and cooling rate on post-thaw sperm quality of ram semen. Aims: The current study aimed to assess the combined impact of cooling rates and pre-freezing equilibration times on post-thaw sperm quality in Kail ram semen (n = 5). Methods: Semen collection was performed using an artificial vagina at 42°C. The pooled semen was divided into equal aliquots and subjected to either slow cooling (SC, −0.27°C/min) or moderate cooling (MC, −0.36°C/min) rates, transitioning from 37°C to 4°C. Equilibration times of 0, 4, 8, and 12 h were employed before freezing. Key results: Semen samples undergoing the SC rate and equilibrated for 4 h exhibited higher (P < 0.05) percentages of progressive motile (PM), rapid progressive (RP), and medium progressive (MP) sperm compared with the MC rate. However, total motility remained unaffected by the cooling rate (P < 0.05). Semen equilibrated for 4 h demonstrated higher (P < 0.05) percentages of PM and RP sperm, as well as improved kinematics (curvilinear velocity, average path velocity, and straight-line velocity) compared with other equilibration times. Nevertheless, equilibration time had no (P > 0.05) impact on the amplitude of the lateral head displacement for semen samples subjected to the MC rate. Notably, the cooling rate did not affect post-thaw sperm kinematics, plasma membrane integrity, or live-sperm percentage (P > 0.05). Semen samples equilibrated for 4, 8, and 12 h exhibited a higher (P < 0.05) percentage of sperm with intact plasma membrane and viability than did those equilibrated for 0 h. Conclusions: In conclusion, slow cooling rate and a 4 h equilibration period were shown to be optimal for preserving post-thaw sperm quality in Kail rams. Implications: The findings highlighted the combined effect of cooling rate and equilibration time on post-thaw sperm quality for optimising sperm cryopreservation protocols in the context of ram semen. The conflicting results regarding the impact of equilibration time on sperm quality warrant further research to improve the post-thaw sperm quality of ram semen. The findings of the present study indicated that slow cooling and equilibration time of 4 h better-preserved sperm quality. The optimisation of sperm-freezing protocol will be beneficial for the widespread use of artificial insemination in sheep. [ABSTRACT FROM AUTHOR]

Objective: The Hu sheep is a renowned breed known for its high reproductive rate. It is in estrus all year round, and its breeding population is gradually expanding. However, the current techniques for cryopreserving semen have limited effectiveness, which hinders the continuous development of this species. The purpose of this study is to explore the effects of different penetrating cryoprotectants (CPAs) and egg yolk (EY) concentrations on the cryopreservation of Hu ram semen to determine the most effective combination. Methods: In this study, the effects of glycerol (GLY), ethylene glycol (EG), dimethylacetamide, dimethyl sulfoxide, different proportions of GLY and EG, EY on sperm quality after thawing were investigated by detecting sperm total motility (TM), progressive motility (PM), straight-line velocity, curvilinear velocity, average path velocity, amplitude of lateral head displacement, wobble movement coefficient, average motion degree, functional integrity (plasma membrane integrity, acrosome integrity) and reactive oxygen species (ROS) level. Results: When GLY and EG were added together, compared to other concentration groups, 6% GLY significantly (p<0.05) increased TM, PM, plasma membrane integrity, and acrosome integrity of thawed sperm. Additionally, it significantly (p<0.05) decreased the ROS level of sperm. In this study, the TM, PM, and membrane integrity of the 6% EG were significantly (p<0.05) higher than those of the control, 1% GLY+5% EG and 6% GLY+6% EG groups. Compared to other concentration groups, 20% EY significantly (p<0.05) improved the TM, PM, and plasma membrane integrity of thawed sperm. However, the integrity of the acrosome increased with the higher concentration of EY. Conclusion: In conclusion, the post-thawed Hu ram semen diluted with a diluent containing 6% GLY and 20% EY exhibited higher quality compared to the other groups. [ABSTRACT FROM AUTHOR]

Goat bucks are seasonal breeders that show variation in sperm quality, endogenous melatonin (MLT), and presumably in the expression of MLT receptors on the sperm throughout the year, which may modify sperm freezability. The aim of this study was to determine whether sperm freezability is associated with (i) endogenous melatonin levels in seminal plasma and (ii) the expression of sperm plasma membrane melatonin receptors (MT1, MT2). To evaluate this, spermatozoa from seven Saanen goat bucks were cryopreserved throughout the year in Mexico using a standard freezing protocol. Seminal plasma MLT concentrations were determined by ELISA and the expression and localization of MT1 and MT2 were detected by immunocytochemistry and confirmed by western blotting. The recovery rate of progressive motility after thawing was higher in spring than autumn and winter; in contrast, the F pattern (CTC assay) was higher in winter than in the other seasons. A proportional increase in the AR pattern (CTC assay) was smaller in winter than in the other seasons and the proportion of sperm showing high plasma membrane fluidity was higher in spring than in summer and autumn. The seminal plasma MLT concentrations showed no significant interseasonal differences. The MT1 receptor was immunolocalised at the apical region of the sperm head, while MT2 was mainly localised in the neck. The relative expression of MLT receptors showed significant differences between summer and winter for all bands, except at 75 kDa of MT2. In conclusion, there was an association between the relative expression of MT1 and MT2 receptors throughout the year and sperm freezability in goat bucks in México. Post-thaw sperm quality is enhanced in semen samples collected during breeding season. • MLT concentration in goats' seminal plasma does not show interseasonal variation. • Expression of MLT receptors in goat spermatozoa shows variation throughout the year. • Goat buck sperm freezability is associated with the expression of MLT receptors. [ABSTRACT FROM AUTHOR]

Females' ability to select mates that can fertilize eggs directly impacts their fitness, but the secondary sexual traits that often guide mate choice may not be directly correlated with male fertility. In externally fertilizing species, such as many fish species, chemical cues in ejaculates may allow females to assess male fertilization capacity. However, chemical cues associated with sperm release have only been described in a few species, and evidence that females use these cues to discriminate among males remains limited. Female sea lamprey, Petromyzon marinus , use several chemical cues to find and interact with potential mates, including a sex pheromone released by males across their gill epithelia and a second sex pheromone released in their semen. Unlike males of many fish species that control pheromone release via urinary pulses, male sea lamprey continually broadcast the gill pheromone once they are sexually mature. This presents a potentially costly scenario for females because attraction to males that are not actively spawning or that are sperm-depleted likely wastes time and energy. We tested the hypothesis that female sea lamprey use pheromones present in seminal plasma to discriminate among potential mates. In-stream behavioural assays revealed that females (1) preferred seminal plasma + male-conditioned water (containing gill-released pheromones) over male-conditioned water alone, (2) preferred male-conditioned water + 2× seminal plasma over male-conditioned water + 1× seminal plasma and (3) oriented towards seminal plasma over large distances and when no male-conditioned water was applied. Taken together, our results indicate that female sea lamprey use seminal pheromones to find and remain near males with immediate fertilization capacity, thereby reducing the risk of wasting time and energy during their single, reproductive window. Finally, our study highlights the importance of multiple cues in mate choice. • We tested whether seminal pheromones provide cues to female lamprey about male status. • Females discriminated males based on the presence of seminal pheromones. • Females discriminated males based on the concentration of seminal pheromones. • Seminal pheromones are a potent, independent cue in sea lamprey. [ABSTRACT FROM AUTHOR]

With a prospect to improve goat artificial insemination, this study aims to evaluate the effects of storage time, egg yolk concentration and semen washing on the rate of motile spermatozoa (spz) in Arbia breed bucks semen stored at 5 °C for 48 h. Semen was collected from three mature bucks using an artificial vagina during December 2018. The ejaculates used in this study (rate of motile sperm > 75 %) were divided into two fractions: A, and B, (unwashed and washed samples, respectively): themselves divided into three fractions, and diluted in Tris-Acid citric based extenders, containing egg yolk at 2 %, 10 % and 20 %. ANOVA test was performed using the R software version 3.3.0. The rate of motile spermatozoa was evaluated after 24 and 48 hours of storage in the refrigerator at 5 °C. It registered a significant decrease as a function of the storage time in the different extenders. It varied between 42.43 ± 9.86 % and 61.29 ± 5.44 %, after 24 hours of storage, and between 35.86 ± 13.95 %, and 55.45 ± 11.65 % after 48 hours of storage. The semen washing significantly reduced the rate of motile spermatozoa in the different extenders. Increasing the concentration of egg yolk in the extender increased the rate of motile spermatozoa, especially during the first 24 hours of storage. In conclusion, the rate of spermatozoa motility of unwashed semen stored at 5 °C is higher in extenders containing high egg-yolk concentration, and decrease according to conservation time. [ABSTRACT FROM AUTHOR]

Objectives: This study aimed to investigate the correlation between testicular shear wave elastography (SWE) values and semen analysis results in men with infertility. Methods: This was a retrospective case–control study. Patients were categorized as normal, abnormal, or azoospermic based on sperm analysis results. Testicular volume was measured using B‐mode ultrasonography using the Lambert formula. Subsequently, 40–80 regions of interest measuring 1.5 × 1.5 mm were manually positioned in both testicles based on their size, and two‐dimensional SWE was applied through virtual touch imaging quantification software. Results: The patients had a mean age of 33.79 ± 6.3 years, with semen analysis revealing normal results in 15 patients (22.4%), pathological findings in 35 patients (52.2%), and azoospermia in 17 patients (25.4%). Right, left, total, and mean testicular volumes were significantly lower in patients with azoospermia compared to those in both normal and impaired semen parameters (P <.05). Conversely, testicular elastography scores were higher in patients with azoospermia than in the other groups (P <.05). The significant negative correlation between volume and elastographic findings remained independent of age (r = 0.4, P <.001). The accuracy rates for detecting impaired semen parameters and azoospermia were 94.3% and 94.1%, respectively, after considering factors such as age, testicular volume (right/left/total), and elastography (right/left/total). Notably, the total mean elastography score ranked first, with 100% in the independent normalized importance distribution of these variables. Conclusion: SWE can be used effectively alone or in combination with other diagnostic tools to evaluate histopathological changes in the testicles of male patients with infertility. [ABSTRACT FROM AUTHOR]

Background: Though a certain level of free radicals is required for the proper maturation of a sperm, excessive level of free radicals is believed to be a major factor contributing to male infertility. Aims and Objectives: This study aimed to identify the level of oxidative stress (OS) among infertile males. Materials and Methods: In the present study, semen Malondialdehyde (MDA) level, as an OS marker was studied in 96 normozoospermics who were taken as controls, and the values were compared separately to semen MDA levels of 75 oligozoospermics, 36 azoospermics and 19 necrozoospermics using unpaired t-test. P < 0.05 was considered statistically significant. Results: The study showed significantly higher semen MDA levels in oligozoospermics and necrozoospermics compared to normozoospermics, but azoospermics did not show a statistically significant difference. Conclusion: We concluded that excessive OS can be one of the major contributing factors for male infertility. [ABSTRACT FROM AUTHOR]

SUMMARY: Good semen quality is a vital tool for the rapid improvement of fertility by allowing maximum use of genetically improved roosters. Evaluation of the best quality semen for different chicken breeds increases artificial insemination efficiency and helps to improve chicken production through the wider usage of genetically superior roosters. The main aim of this systematic review is to summarise the fertility responses and semen quality parameters of different poultry strains to provide a summarised clue for breeders working on indigenous poultry breeds. There was a difference in semen volume per ejaculation between breeds' fertility; however, there was similar semen volume for both Bar Plymouth Rock and Sasso breeds (0.8 mL). The semen volume of Bar Plymouth Rock ranged from 0.3 to 0.8 mL. There was a maximum (86%) of sperm motility in indigenous chickens from Chung University and a lower motility for indigenous chickens at the University of Urmia (29.6%). Viable sperm for Venda chicken was higher (96.8%) than the other breeds, and its performance ranged from 46.7% to 96.8%; however, there was the lowest (13%) sperm viability for Hubbard rooster. The sperm viability for White Leghorn also ranged from 49.7% to 82.5%. The abnormality of sperm was higher (72.6%) for the Rhode Island Red breed but lowest (3.2%) for indigenous chicken from Taiwan and 7% for Dahlem Red. Turkey's breed was superior for spermatozoa concentration (10.6 × 109/mL) than other rooster breeds. The lowest (1.2 × 109/mL) sperm concentration was reported for Beijing chickens. In terms of the fertility rate, there was a higher (93%) fertility rate for White Leghorn (ranging from 81.7% to 93%) and the lowest (50.6%) fertility rate for South African Naked Neck (ranging from 50.6% to 55.7%. The observed difference in fertility and semen characteristics across the breeds was mainly attributed to the genetic, age, nutrition type, management difference, production type, agroecology difference, collection frequency and collection interval. [ABSTRACT FROM AUTHOR]

Sperm oxidative stress has been extensively associated to male infertility. However, tests to detect this parameter have not been yet introduced in clinical practice and no definitive data are present on the extent of oxidative stress in male infertility. In this study, we used a novel and reliable flow cytometric method to reveal sperm ROS production in subfertile patients (n = 131) and in healthy donors (n = 31). Oxidative stress was higher in subfertile patients (14.22 [10.21–22.08]%) than in healthy donors (9.75 [8.00–14.90]% (p < 0.01)), but no correlation was found with age, semen quality or sDF. We also failed to detect an increase in sperm ROS production with semen viscosity or leukocytospermia, but a sharp impact of semen bacteria was evident (with bacteria: 31.61 [14.08–46.78]% vs. without bacteria: 14.20 [10.12–22.00]%, p < 0.01). Finally, after establishing a threshold as the 95th percentile in healthy donors, we found that 29% of subfertile patients exceeded this threshold. The percentage decreased to 25.56% when we excluded subjects with bacteriospermia and increased to 60.87% when only these patients were considered. In conclusion, 29% of subfertile patients showed an excessive sperm ROS production. Surprisingly, this parameter appears to be independent from routine semen analysis and even sDF determination, promising to provide additional information on male infertility. [ABSTRACT FROM AUTHOR]

Background: Historically, viruses have impaired the functionality of human systems. The discovery of novel viruses and the resurgence of established strains heighten concerns about their implications for reproduction. In the aftermath of COVID-19, research efforts have intensified to broaden the understanding of the severe acute respiratory syndrome coronavirus-2's (SARS-CoV-2) effect on male reproduction across diverse populations. Various findings have been reported, with more studies highlighting the adverse impact of SARS-CoV-2 on semen parameters and, consequently, male fertility. This review aims to comprehensively assess and consolidate existing evidence on the impact of SARS-CoV-2 on semen quality and male fertility. It highlights the potential mechanism of action and further underscores the implications for assisted reproductive technology. Methodology: A thorough literature search was conducted across various electronic databases, including PubMed, Scopus, Google Scholar, Embase, and Web of Science. Studies published between January 2020 and May 2024 were included if they explored the impact of SARS-CoV-2 on semen quality. Results: Twenty-nine (29) studies were included in the review. These studies varied in findings but delineated a pattern and trend. While most studies noted a decline in sperm parameters—motility, count, concentration—altered morphology, hormonal imbalances, and increased DNA fragmentation in COVID-19 patients, others reported normal semen parameters 3 to 6 months post-recovery. Few studies reported no change in semen parameters, especially with mild disease. Potential mechanisms underscoring these effects include the presence of fever and consequent release of pro-inflammatory cytokines—interleukin 1β, tumor necrosis factor (TNF), and interferon-gamma (IFNγ). In addition, the activities of the angiotensin-converting enzyme 2 (ACE2) and the transmembrane serine protease 2 (TMPRSS2) receptors have been implicated as gateway mechanisms for viral entry. The long-term consequences and comparisons with other viral infections highlight the complexity of drawing definite conclusions. The different findings on semen changes have implications for assisted reproduction and family planning. Research suggests potential negative effects on gonadal function, emphasizing the need for long-term follow-up studies to understand the persistent effects on male fertility biomarkers. Conclusion: A multidisciplinary approach is essential to optimizing male reproductive health during and after SARS-CoV-2 infection. This includes incorporating assessments into vaccine safety studies to address fertility concerns. [ABSTRACT FROM AUTHOR]

Infertility is an inability to conceive after a reasonable period of time (12 months) without the use of contraception or due to a person’s incapacity to procreate, whether independently or with a spouse. Problems with the production and maturation of sperm are the most common causes of male infertility additionally; the motility is the major functional character that determines the fertilizing ability of spermatozoa. Therefore the goal of this study is to get better certain sperm function parameters in vitro of asthenozoospermic patient. The World Health Organization (WHO) and many studies considered the infertility as a disease and so many couples complaining from unsuccessful assisted reproductive technologies (ART) procedures to overcome their problem. The goal of this study is to improve certain sperm function feature in vitro of asthenozoospermic semen patients by using combination of motility inducing namely; Maca, L-carnitine and Pentoxifylline that enhance the medium to improve certain sperm characters that might be utilized for ART centers. Semen aliquots were collected from ninety patients with asthenozoospermia who participated in present study, the volume of semen samples with normal ejaculate when was ranged between 1.4-6ml and can be measured by using a measure pipette or conical graduated tube; Inclusion criteria was Asthenozoospermia, oligozoospermia and teratozoospermia men, Infertile idiopathic men also, fertile normozoospermic men. While Exclusion criteria was Azoospermic men, Alcoholic, Patients under treatment with antibiotics and men with Varicocele. The samples split into two equal groups at random. Using Ham’s F12 medium, one portion served as the control group, and the other was the treatment group, which was mixing by combining the following ingredients, Maca powder extracts (Lepidium meyenii) (M) 1 mg/ml, 0.5 mg/ml of L-Carnitine (LC), and 10 mg/ml of Pentoxifylline (PTX). The data were analyzed using Statistical Package for Social Sciences (SPSS) version 23.0. The descriptive statistics including frequency, range, mean and standard error, Data from treated and control groups were expressed as mean ± SEM and to compare value between experimental and control groups using Students t-test. Layering approach is used to investigate sperm parameters before and after in vitro activation. The information showed a very large (p< 0.001) increase in active sperm motility grade A, percentage of progressive motility with significant increase in morphologically normal sperm (MNS) with decreased in DNA fragmentation index after activation by layering technique with novel combination medium compared to Hams F12 medium and before activation. The present work stated that novel combination medium (LC, maca and PXT) have potential effects to improve sperm characters in male infertility factors and suggested to be used for sperm preparation and activation in ART programs. [ABSTRACT FROM AUTHOR]

Background: Male Genital Schistosomiasis (MGS) remains an often-overlooked chronic sequela of urogenital schistosomiasis in endemic areas of sub-Saharan Africa. As part of a 2-year longitudinal study on Hybridization of UroGenital Schistosomiasis (HUGS) in Malawi, a MGS sub-study was conducted to assess whether hybrid schistosomes were incriminated. Methods: During recruitment, demographic, health and socio-economic data were collected through individual questionnaire interviews in Mthawira community from Nsanje District along Shire River and Samama community from Mangochi District along Lake Malawi shoreline. Urine and semen samples were collected and analysed to determine the identity of schistosome infection. Urine filtration and microscopy, direct microscopy of semen and its sediments (after centrifugation) were performed. Thereafter, the sediments were examined by molecular DNA analysis with a novel two-tube real-time PCR assay. The participants were also screened for Human papilloma virus (HPV) and other sexually transmitted infections (STIs). Results: Twenty-two men were recruited for the sub-study, 8 in Nsanje District and 14 in Mangochi District, with a median age of 22.0 years. By microscopy, ten (45.7%) participants had Schistosoma ova in their urine, 11 (50.0%) in semen while 16 (72.7%) were positive by real-time PCR. One participant had both S. haematobium and S. mattheei ova in his semen, three showed symptoms, and one had a mixed infection of S. mansoni and possible S. haematobium-S. mattheei hybrid. Twelve men had detectable high-risk HPV serotypes 16, 18 and others while six had Trichomonas vaginalis and other STIs. Conclusion: Zoonotic and hybrid schistosomes can cause MGS similar to human schistosomes, which can be co-infected with HPV and STIs, thereby posing a new challenge in diagnosis, management and control measures in resource poor settings. Increased awareness of these infections among local communities and primary healthcare workers and improvement of disease management are needed and advocated. [ABSTRACT FROM AUTHOR]

Simple Summary: In this study, semen collection techniques, in particular, Urethral Catheterization after Pharmacological Induction (Ur.Ca.P.I.) and electroejaculation, have been applied in two tigers and two leopards. Information about sperm collection in tigers and leopards has been reported, describing the authors' experience and presenting new data about a topic difficult to study due to the animal species involved. In particular, collection was possible in both species thanks to the use of electroejaculation. Ur.Ca.P.I. gave positive results in leopards, while in tigers, this technique did not allow satisfactory results. Assisted reproduction techniques play a significant role in veterinary medicine, and although they are widely used in domestic animals, they are also becoming increasingly relevant in clinical practice for wild felids, especially in the conservation efforts for endangered species. In this study, the result of two semen collection techniques, urethral catheterization after pharmacological induction (Ur.Ca.P.I.) and electroejaculation, are described, aiming to provide new practical information about sperm collection using the Ur.Ca.P.I. technique and electroejaculation in tigers and leopards, describing the authors' experience and presenting new data and observations. The following descriptive study included two subjects of Panthera tigris species and two of Panthera pardus. These subjects, after general anesthesia, underwent sperm collection initially with Ur.Ca.P.I. and, subsequently, with electroejaculation. Sampling was made possible in both species thanks to the use of electroejaculation. Sperm volumes in leopards ranged from 0.3 to 0.5 mL and in tigers from 0.5 to 2.177 mL. Sperm concentration in leopards ranged from 136 × 106 to 280 × 106 sperm/mL, and in tigers, from 21.5 × 106 to 354 × 106 sperm/mL. Urethral catheterization gave positive results in leopards, with sperm volumes ranging from 25 up to 150 µL and a concentration ranging from 110 × 106 up to 1082 × 106 sperm/mL. In tigers, unlike in leopards, the use of the Ur.Ca.P.I. technique encountered difficulties that did not allow satisfactory results to be obtained. Therefore, it would be useful to test the feasibility of urethral catheterization on a larger group of individuals in order to have more meaningful feedback. Finally, because electroejaculation always allowed semen collection in tigers, with a higher sperm quality than samples collected by Ur.Ca.P.I., we currently consider it the technique of choice for the collection of semen material in this species. [ABSTRACT FROM AUTHOR]

Semen persicae is the dried mature seeds of Prunus persica (L.) Batsch and P. davidiana (Carr.) Franch and is commonly used in traditional Chinese medicine (TCM) formulations because of its variety of biological effects. The present study aimed to evaluate the antioxidant activity and toxicity profiles of semen persicae extract (SPE) after determining the amygdalin content (4.95%) using HPLC. Regarding the in vitro antioxidant activity, SPE with 2 mg/mL concentration scavenged 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl, and ABTS free radicals with rates of 51.78%, 55.47%, and 57.16%, respectively. The same concentration of SPE chelated 30.76% Fe2+. The in vitro cytotoxicity study revealed that SPE induced 92.45% cell viabilities of HEPG2 even at 2000 μg/mL. In the acute toxicity study, oral administration of SPE did not provoke mortality or any toxic signs at doses up to 2000 mg/kg bw. Repeated oral administration for 28 days at doses of 100, 300, and 600 mg/kg per day in rats did not show any toxicity signs or gross pathological abnormalities. The results of the present research provide basic reference data for SPE with a moderate effect on antioxidant activity and low toxicity for future screening of biological and pharmacological properties. [ABSTRACT FROM AUTHOR]