Beneficial effects of oral antioxidant supplementation on semen quality parameters, reproductive hormones, and sperm DNA integrity in men with idiopathic oligoasthenoteratozoospermia.

OBJECTIVE: Recently, oral antioxidants in combined forms have been used to treat men with idiopathic infertility. This study aimed to evaluate the effects of treatment with vitamin C, vitamin E, selenium, zinc, arginine, L-carnitine, and coenzyme Q10 on sperm quality parameters, DNA integrity, reproductive hormones, and pregnancy rates in men with infertility and idiopathic oligoasthenoteratozoospermia (OAT). METHODS: A prospective study was conducted on 420 men with infertility and idiopathic OAT who took an oral supplement of antioxidant SP-Power tablets twice daily for 6 months. Semen quality, reproductive hormones, and the DNA fragmentation index (DFI) were evaluated at baseline and at 3 and 6 months after supplementation, using the World Health Organization 2021 guidelines. RESULTS: No significant difference was observed in volume or the percentage of typical morphology during treatment. A significant improvement in sperm concentration was observed after supplementation (8.67±1.41, 12.17±1.91, and 19.01±0.86 at baseline, 3, and 6 months respectively, p<0.01). The total motility, progressive motility, and total motile sperm count also increased significantly (p<0.01), whereas the DFI decreased after 6 months. There was an increase in normal FSH levels and testosterone levels after 6 months of supplementation of antioxidant SP-Power but these differences were not statistically significant (p=not significant and p=0.06, respectively). CONCLUSION: Supplementation with SP-Power tablets improved sperm quality parameters, sperm DFI, some reproductive hormones, and pregnancy rates in men with infertility and idiopathic OAT, which could be attributed to the supplement's synergistic antioxidant action. Further studies are needed to determine the effects of supplementation on oxidative stress markers.

Effect of nutraceutical supplementation on semen quality in stallions.

BACKGROUND: The use of reproductive biotechnologies in equine practice has shown that some stallions are subfertile, so ways to improve fertility have been sought. OBJECTIVE: This study aimed to evaluate the effect of nutraceutical supplementation on improving semen quality in Quarter Horse stallions. METHODS: Semen from six Quarter Horse stallions was assessed for 4 months every 20 days using the computer-assisted semen analysis system. They were evaluated for 60 days before supplementation; then, the same stallions were re-evaluated for 60 days with nutraceutical supplementation (30 g/day). RESULTS: Volume showed no significant difference (p > 0.05) with nutraceuticals. Sperm concentration (10x6 ) was significantly higher with supplementation (339.4 ± 17.5 sperm/mL) than without supplementation (224.6 ± 19.9). Sperm abnormalities (%) were significantly (p < 0.05) lower with supplementation (14.3 ± 0.6) than without supplementation (19.1 ± 0.4). Sperm kinematic parameters, total motility (TM), progressive motility (PM), rectilinear velocity (VSL), the velocity of the trajectory (VAP) and curvilinear velocity (VCL), were significantly better with supplementation (p < 0.05). CONCLUSIONS: Based on the results, it is concluded that nutraceutical supplementation improved semen parameters in Quarter Horse stallions.

Lipids in the reproduction of sires.

The main class of nutritional interest for lipids are fatty acids (FA), which correspond to 90% of triglycerides, the main form of lipid storage in both plants and animals. FAs serve as a source of energy in the diet of cattle; however, they also have an important non-caloric effect on animal organisms as they are important components of the physical and functional structures of cells and participate in the composition of steroid hormones. As such, research has studied the improvement of semen quality through the provision of polyunsaturated FAs in bull diets, as well as the use of FAs in semen extenders in order to reduce damage to sperm cells, which can alter lipid composition and the quality of frozen sperm. Therefore, the objective of this work was to review the effectiveness of lipids on reproductive efficiency, based on their effects on semen quality and hormonal production. Supplementation with polyunsaturated FAs positively alters semen composition and in vitro fertility; however, results vary according to the type of FA used, the method of administration, and its quality. Fish oil and linseed oil showed better results in qualitative parameters in fresh and thawed semen. The use of cyclodextrins to incorporate or extract cholesterol from plasma membranes can also improve the viability of cryopreserved semen.

Influence of spirulina supplementation on growth performance, puberty traits, blood metabolites, testosterone concentrations, and semen quality in Barki male lambs.

Background: The fertility and genetic value of the flock can be enhanced by selecting lambs with highly developed early puberty characteristics. Spirulina (SP) has been evaluated as a natural product supplement to boost lamb growth, immunity, and productivity. Aim: Study growth performance, blood metabolites, puberty development traits, semen quality, and seminal plasma biochemical concentrations of growing Barki lambs when supplemented with SP at different levels. Methods: in a 24 weeks study, 30 Barki male lambs weighing an average of 21.78 ± 2.56 kg, with a body condition score of 3.20 ± 0.55 and an age of about 16 ± 0.24 weeks were used. The lambs were randomly assigned to three groups (10 lambs each) of daily SP supplementation levels per lamb of 0 ml (control), 50 ml (SP1), and 100 ml (SP2). The SP powder was made into a water suspension using SP to water ratio of 1 g:10 ml. The growth characteristics, as well as the development of puberty, blood metabolites, and semen quality analysis of every lamb, were measured. Results: The growth performance was greater (p < 0.05) in SP2 lambs compared with other lambs. While daily dry matter intake was not affected by SP treatment, feed efficiency had significantly improved in SP2 groups. Furthermore, the SP2 lambs have attained puberty at early ages than the control lambs. The testes volume of SP2 lambs was bigger (p < 0.05) than other groups throughout the pre-pubertal up to the puberty stage. The addition of SP had no effects on the total protein, glucose, and triglycerides concentrations. Meanwhile, the cholesterol concentration was lowest (p < 0.05) in the SP2 lambs. The blood and seminal plasma levels of alanine aminotransferase and aspartate aminotransferase decreased (p < 0.05) in the SP lambs more than their control counterparts. The levels of superoxide dismutase reduced glutathione, and total antioxidants had increased (p < 0.05) in the treated lambs compared with the control group. Further, the malondialdehyde levels decreased (p < 0.05) in the SP-treated lambs. Additionally, the SP2 lambs produced better semen quality than the control lambs. Conclusion: SP supplementation (100 ml/head/day) enhanced growth performance, feed efficiency, and antioxidative status, exerting a positive influence on the physiological parameters and sexual behavioral patterns at puberty in Barki lambs.

Transcriptomic analysis of mechanism underlying the effect of induced molting on semen quality and reproductive performance in aged Houdan roosters.

The reproductive performance of breeder roosters has significant economic importance in the poultry industry. Breeder roosters have severely reduced semen quality with age and will be at risk of culling in the following years. In order to extend the use of breeder roosters, we drew on the induced molting model of hens and selected 35 Houdan roosters aged 50 wk for induced molting. By comparing the body weight, testicular weight, semen quality, and reproductive performance before and after induced molting, we found that induced molting could restore the body weight and testicular weight to the levels before molting (P > 0.05). At the same time, it significantly improved sperm motility (P < 0.05) and also improved reproductive performance such as fertilization rate and hatching rate. To further reveal the mechanism underlying the effects of induced molting on semen quality and reproductive performance in aged Houdan roosters, we collected testes from 3 periods: 1 d before fasting (F0), 15 d after fasting (F15), and 32 d after recovery feeding (R32) for transcriptome sequencing analysis. A total of 5,671 genes were detected in F0, F15, and R32, and trend analysis of the 5,671 differential genes showed 2 significant trends (profile 5 and profile 2). KEGG enrichment analysis of the genes in the 2 profiles, revealed significantly enriched pathway regulation of actin cytoskeleton. In the regulation of actin cytoskeleton pathway, we found a protein kinase gene (SRC) and a senescence gene (ROCK2). SRC was highly expressed at F15, leading to the phosphorylation of key substrates, which in turn disrupted the Sertoli cell spermatid connection and the spermiogenesis process, resulting in no mature spermatozoa produced from F15, SRC expression was inhibited at R32, the expression level was reduced, and mature spermatozoa reappeared. The senescence gene ROCK2 was highly expressed at F15 compared to F0 and R32, which may have been responsible for inducing senescence atrophy in the testes.

Effect of In Vitro Addition of Melatonin and Glutathione on Seminal Parameters of Rams in Diluted Semen and after Thawing.

The present study aimed to evaluate the effects of melatonin (MLT), glutathione (GSH), and their combination on ram semen quality after thawing. During eight weekly sessions, semen from three Merino rams was pooled, diluted with an egg-yolk-based semen extender, and divided into four groups: control, 1 mM MLT, 5 mM GSH, and 1 mM MLT + 5 mM GSH. Diluted semen was evaluated before and after the freezing process. The supplementation of diluted semen with GSH at 5 mM had a deleterious effect on total motility progressive (TPM), curvilinear velocity (VCL), straight-line velocity (VSL), average-path velocity (VAP), linearity (LIN), and straightness (STR) and increased slow spermatozoa (%). MLT at 1 mM also had a negative effect on TPM, VSL, and STR in diluted semen. In thawed semen, 1 mM MLT increased the TPM compared with the control group. VSL was lower in the 5 mM GSH group than in the 1 mM MLT group. Additionally, the combination of both antioxidants attenuated the negative effect of 5 mM GSH on TPM, VSL, and BCF. These results indicate that 5 mM GSH impairs or does not improve sperm kinetic parameters in either diluted or thawed semen. They also suggest that MLT combined with GSH plays a protective role against these effects.

Reducing oxidative stress by κ-carrageenan and C60HyFn: The post-thaw quality and antioxidant status of Azari water buffalo bull semen.

Azeri water buffalo is a species of great interest due to the high quality of its products such as milk. Due to the decreasing trend of its number and risk of extinction in the future, our attention is directed towards ensuring the preservation of its genetic reserves by keeping its sperm. Using antioxidants in semen extender is one of the ways to reduce the detrimental effects of freezing process on post-thawed quality of spermatozoa. This study was conducted to determine the effect of κ-carrageenan (k-CRG) and C60HyFn supplemented semen extender on the quality of post-thawed Azari water buffalo spermatozoa. A total of 30 semen samples were obtained from three buffaloes using an artificial vagina (twice a week for five weeks = 10 replicates). The samples (n = 3) from each replicate were pooled and divided into equal aliquots to prepare 14 extender groups, including control (C), k-0.2, K-0.4, K-0.6, K-0.8 (containing 0.2, 0.4, 0.8 mg K-CRG/mL, respectively), C-0.1, C-0.2, C-0.4, C-0.8, C-1, C-5, C-10, C-20, and C-40 (containing 0.1, 0.2, 0.4, 0.6, 0.8, 1, 5, 10, 20, 40 µM C60HyFn, respectively), and then frozen. After thawing, motility and velocity parameters, plasma membrane integrity (PMI) and functionality (PMF), DNA damage, Hypo-osmotic swelling (HOS) test, malondialdehyde (MDA), total antioxidant capacity (TAC), glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase glutathione activities and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging were evaluated. In vivo fertility was compared between k-0.6, C-1 and control groups. 60 buffalo were inseminated 24 h after the onset of estrus. The diagnosis of pregnancy was performed rectally at least 60 days after fertilization. Total and progressive motility and velocity parameters were improved by k-0.4, k-0.6, k-0.8, C-0.4, C-0.8, C-1, C-5, and C-10 groups) compared to the other groups. Plasma membranes integrity and PMF were improved by k-0.4, k-0.6, C-0.4, C-0.8, C-1, C-5, and C-10 groups compared to other groups, while in terms of sperm DNA damage K-0.4, K-0.6, K-0.8, C-0.2, C-0.4, C-0.8, C-1, C-5, and C-10 groups showed better results compared to the control group. The evidence also showed that k- 0.4, k-0.6, k-0.8, C-0.4, C-0.8, C-1, C-5, and C-10 groups could improve TAC, and decrease MDA levels. Also, k-0.4, k-0.6, k-0.8, C-0.2, C-0.4, C-0.8, C-1, C-5, and C-10 groups could improve GPx, CAT, and GSH levels, but no significant difference was found regarding SOD compared to the other groups. DPPH scavengers were tested by K-0.6, K-0.8 and C-1, C-5, C-10, C-0.8, C-0.4 and C-0.2 groups and compared to other groups improved. The fertility rate [70% (14/20)] was higher in C-1 than other groups. To conclude that k-CRG and C60HyFn supplementation can increase the quality parameters of cryopreserved buffalo semen after thawing and that 1 M C60HyFn can increase in vivo fertility of buffalo semen.

Seasonal Variations in Semen Quality, Testosterone Levels, and Scrotal Size following Dietary Flaxseed Oil and Ascorbic Acid in South African Indigenous Rams.

The purpose of this study was to determine the seasonal variations in semen quality, testosterone levels, and scrotal size, following dietary flaxseed oil and ascorbic acid in South African indigenous rams. A total of 22 South African indigenous rams were randomly distributed into five treatment diets from June 2021 to May 2022 (12 months). To allow for the spermatogenesis period, semen was collected after sixty days of dietary supplementation with treatment diets. Blood was collected twice a week using an 18-gauge needle and vacutainer tubes and sent to the laboratory for testosterone analysis. Semen and blood collection were repeated eight times each season. The scrotal size (circumference, length, and width) was measured using a flexible measuring tape. Data was subjected to the General Linear Model (GLM) in Minitab® 2017. Treatment means were separated using Fisher's t-test and considered significantly different when the p-value was less than 0.05. Seasons and diet had an effect on progression, total motility, and testosterone levels. For instance, NC during the spring season had the lowest progressive motility (42.84 ± 5.32), followed by the summer (49.38 ± 4.49), winter (62.46 ± 4.35), and autumn (63.26 ± 3.58). Notably, when treatment diets were introduced, improvements were realized, and there were significant differences (p < 0.05) among the seasons following supplementation of FLAX, ASCA, and FLAX + ASCA, except for FLAX in the autumn season (53.83 ± 4.16). Total motility did not differ significantly (p > 0.05) between the seasons when the NC and PC diets were supplemented; nevertheless, there was an improvement when FLAX, ASCA, and FLAX + ASCA were supplemented. Testosterone levels were significantly influenced by the seasons when negative and PC diets were supplemented. It is noteworthy that supplementing FLAX + ASCA can reverse the influence of the season on the testosterone levels (spring, 27.52 ± 4.42; summer, 20.23 ± 5.11; autumn, 25.24 ± 3.96; and winter, 25.92 ± 4.42). In conclusion, seasons do affect semen quality and testosterone levels of South African indigenous rams. However, flaxseed oil and ascorbic acid can reverse the seasonal variations in semen quality and testosterone levels.

Low serum anti-Müllerian hormone is associated with semen quality in infertile men and not influenced by vitamin D supplementation.

BACKGROUND: Anti-Müllerian hormone (AMH) is released by testicular Sertoli cells and of great importance during fetal male sexual development, but less is known about the role of circulating AMH during adulthood. In vitro studies have shown that vitamin D may induce AMH transcription, but a controlled trial investigating the possible effect of vitamin D on serum AMH has not been conducted in men. METHODS: A single-center, double-blinded, randomized placebo-controlled clinical trial (NCT01304927) conducted in Copenhagen, Denmark. A total of 307 infertile men were included and randomly assigned (1:1) to a single dose of 300,000 IU cholecalciferol followed by 1400 IU cholecalciferol + 500 mg of calcium daily (n = 151) or placebo (n = 156) for 150 days. Difference in serum AMH was a predefined secondary endpoint. Explorative outcomes were associations between serum AMH and gonadal function in infertile men. The primary endpoint was difference in semen quality and has previously been published. RESULTS: Infertile men in the lowest AMH tertile had significantly lower sperm concentration (∆T3-1 16 mill/mL (228%); P < 0.001), sperm count (∆T3-1 55 million (262%); P < 0.001), motile sperm count (∆T3-1 28 million (255%); P < 0.001), progressive motile sperm count (∆T3-1 18 million (300%); P < 0.001), testis size (∆T3-1 2.7 mL (16%); P < 0.001), serum inhibin B (∆T3-1 72 pg/mL (59%); P < 0.001), inhibin B/FSH ratio (∆T3-1 48 (145%); P < 0.001), and higher FSH (∆T3-1 2.6 (38%); P < 0.001) than the tertile of infertile men with highest serum AMH. Vitamin D supplementation had no effect on serum AMH compared with placebo treatment. CONCLUSIONS: In infertile men, low serum AMH is associated with severely impaired gonadal function illustrated by poor semen quality and lower testosterone/LH ratio. Serum AMH in infertile men was not influenced by vitamin D supplementation.

Sperm mitochondrial DNA copy number mediates the association between seminal plasma selenium concentrations and semen quality among healthy men.

Selenium (Se) is essential for successful male reproduction. However, the association of Se status with human semen quality remains controversial and the underlying mechanisms are poorly understood. We measured seminal plasma Se concentrations, sperm mitochondrial DNA copy number (mtDNAcn), and sperm quality parameters among healthy Chinese men screened as potential sperm donors. Linear mixed-effects models were used to investigate the associations of within-subject pooled seminal plasma Se concentrations (n = 1159) with repeated sperm quality parameters (n = 5617); mediation analyses were applied to evaluate the mediating role of sperm mtDNAcn (n = 989). Seminal plasma Se concentrations were positively associated with sperm concentration and total count (both P for trend < 0.001). In adjusted models, men in the top vs. bottom quartiles of seminal plasma Se concentrations had 70.1 % (95 % CI: 53.3 %, 88.9 %) and 59.1 % (95 % CI: 40.5 %, 80.2 %) higher sperm concentration and total count, respectively. Meanwhile, we observed inverse associations between seminal plasma Se concentrations and sperm mtDNAcn, and between sperm mtDNAcn and sperm motility, concentration, and total count (all P for trend < 0.05). Mediation analyses suggested that sperm mtDNAcn mediated 19.7 % (95 % CI: 15.9 %, 25.3 %) and 23.1 % (95 % CI: 17.4 %, 33.4 %) of the associations between seminal plasma Se concentrations and sperm concentration and total count, respectively. Our findings suggest that Se is essential for male spermatogenesis, potentially by affecting sperm mtDNAcn.

In Vitro and In Vivo Studies on the Efficacy of Zinc-Oxide and Selenium Nanoparticle in Cryopreserved Goat (Capra hircus) Spermatozoa.

Different nanoparticles (NPs) are currently being investigated for their potential role as cryoprotectant during semen cryopreservation in several mammalian species. It may be possible to improve semen quality following cryopreservation by supplementation of NPs in the freezing extenders. The present study was carried out in semen collected from four (4) Assam Hill Goat bucks (10 ejaculates per buck) to investigate the effect of supplementing zinc oxide (ZnO) and selenium (Se) NPs in Tris-citric acid-fructose yolk (TCFY) extender on in vitro sperm quality and in vivo fertility rate after freeze-thawing. The size morphology and zeta potential of ZnO and Se NPs were evaluated prior to its incorporation in the freezing extender. Qualified semen samples (> 70% progressive motility) were divided into five (5) aliquots and then diluted in TCFY extender containing ZnO and Se NP supplementation at different concentrations (T0, control; T1, 0.1 mg/mL ZnO NPs; T2, 0.5 mg/mL ZnO NPs; T3, 0.5 µg/mL Se NPs; and T4, 1 µg/mL Se NPs). Diluted semen was packed in 0.25 mL straws and then stored in liquid nitrogen. After thawing, post-thaw in vitro sperm attributes were evaluated. Finally, the effect of NPs on in vivo fertility rate was checked in heat-synched does (n = 70) by artificial insemination (AI) using straws that showed superior results during the in vitro study. Results showed that ZnO and Se NPs were poly-crystalline in nature with particle size below 100 nm (nm). The evaluated post-thaw sperm in vitro attributes were significantly (p < 0.001) higher in T1 in comparison to T0. The antioxidant enzyme activities were significantly (p < 0.001) higher in T1. Lipid peroxidation (LPO) profile was significantly (p < 0.001) lower in T1. Sperm motility and mitochondrial membrane potential (MMP) had a highly significant (r = 0.580, p < 0.05) association in T1. No significant (p > 0.05) differences in pregnancy rates were recorded after AI in the different treatments. In conclusion, extender supplemented with 0.1 mg/mL ZnO NPs improved post-thaw semen quality of goat spermatozoa consequently by increasing activities of endogenous antioxidant enzymes thereby lowering LPO levels. However, improved in vitro outcomes might not correspond to improved field fertility outcomes.

Determining the Optimal Dosage of Lecithin Nanoliposome in Rooster Semen Freezing Medium and Fertility Potential.

Introduction: Lecithin nanoliposome (nano-LPO), with its cryoprotective properties, is considered to enhance the performance of a traditional semen cryoprotectant. Objective: To determine the optimal dose of lecithin nano-LPO added to the rooster semen extender. Materials and Methods: Semen samples collected weekly from eight broiler breeder roosters were mixed and aliquoted into five equal subsamples, during the five successive weeks. The subsamples were then diluted with a semen extender containing 0%, 0.5%, 1%, 1.5%, or 2% of lecithin nano-LPO. Post-thawed semen quality attributes, including sperm motility and velocity parameters, plasma membrane functionality, mitochondrial membrane potential (MMP), apoptosis-like changes, and fertility potential, were evaluated. Results: Total motility and velocity parameters, including curvilinear velocity (VCL), straight-line velocity (VSL), average path velocity µm/s (VAP), straightness (STR), linearity (LIN), lateral head displacement (ALH), and wobble (WOB) were quadratically (p < 0.01) influenced by graded levels of lecithin nano-LPO, such that the highest values were obtained when 1% of lecithin nano-LPO was used. Treatments had no significant effect on plasma membrane functionality; however, MMP (p < 0.08) and percentages of live and dead spermatozoa (p < 0.05) quadratically responded to increasing levels of lecithin nano-LPO, where the best outcome was found when about 1% of lecithin nano-LPO was used in the semen extender. The percentage of apoptotic spermatozoa cubically responded to increasing levels of lecithin nano-LPO (p ≤ 0.07). No significant trend of fertility rate was found in response to addition of lecithin nano-LPO levels. Conclusions: Supplementing an extender with 1.10% of lecithin nano-LPO is shown to be the optimal dose associated with the most improvement in post-thawed rooster sperm velocity measurements.

Dietary supplementation of spirulina and canthaxanthin boosts laying performance, lipid profile in blood and egg yolk, hatchability, and semen quality of chickens.

This study examined the impacts of dietary spirulina (SP) and canthaxanthin (CX) supplementation to laying hens on reproductive performance and cock's semen quality evaluation. Three hundred and sixty of indigenous Sinai chickens (300 hens + 60 cocks), 48-weeks-old, were randomly divided into five treatments; a control and four supplemented (0.6 g SP mg/kg, 0.8 g SP mg/kg, 6 mg CX/kg diet, and 8 mg CX/kg diet) with six replications of 10 females and 1 male per treatment for laying trial and 1 male for semen evaluation. The animals were fed with experimental diets for 12 weeks. Supplementing 6 mg CX/kg diet led to an increase in the final body weight and egg production traits, while 8 mg CX/kg diet resulted in an improving feed conversion ratio. Supplementation of 0.6 g SP mg/kg or 6.0 mg CX/kg diet resulted in the highest egg production. The heaviest egg weight and egg mass were induced by 0.8 g SP mg/kg. Eggs laid from hens fed 0.6 g SP/kg diet had a higher concentration of cholesterol than those fed 0.8 g SP/kg and those fed 6.0 and 8.0 mg CX/kg. Cocks fed 6 mg CX/kg and those fed 0.8 g SP/kg diet had the greatest sperm concentration. The supplementation of 0.8 g SP mg/kg diet or both 6 and 8 mg CX/kg diet enhanced hatchability. So, dietary spirulina (0.8 mg/kg) and canthaxanthin (6 mg/kg) supplementation to Sinai laying hens and cocks could be used to improve productive and reproductive and performance.

Identification of potential candidate genes and regulatory pathways related to reproductive capacity in hypothalamus and pituitarium of male ducks (Anas platyrhynchos) by differential transcriptome analysis.

The improvement of reproductive capacity of poultry is important for the poultry industry. The existing studies on reproductive capacity mainly focus on the testis tissue, but few reports on regulationary effect of brain neuroendocrime on reproductive capacity have been available. The hypothalamus-pituitarium-gonad (HPG) axis is an important pathway regulating spermatogenesis and sexual behavior. This study analyzed the gene expression in the hypothalamus and pituitary tissues of male ducks in high-semen-quality group (DH), low-semen-quality group (DL), and non-response group (DN) by RNA-sequencing. A total of 1980 differentially expressed genes (DEGs) were identified, and significantly less DEGs were found in pituitary gland than in hypothalamus. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed that these DEGs were mainly enriched in nerve-related and synapse-related biological processes, mitochondrial inner membrane formation pathway, and ribosome structure pathway. Notably, the neuroactive ligand-receptor interaction pathway significantly enriched in all three comparisons (DH vs. DL, DH vs. DN, and DL vs. DN) was related to different reproductive performance such as semen quality and sexual response. Furthermore, six genes, including POMC, CPLX2, HAPLN2, EGR4, TOX3, and MSH4, were identified as candidate genes regulating reproductive capacity. Our findings provide new insights into the regulation mechanisms underlying the reproductive performance of male poultry, and offer a valuable reference for duck breeding programs aimed at promoting reproductive capacity.

Individual reproductive capacity is crucial to poultry breeding and reproduction. The hypothalamus­pituitarium­gonad (HPG) axis is an important pathway regulating animal spermatogenesis and sexual behavior. This study identified the neuroactive ligand­receptor interaction pathway as the potential biological pathway regulating the semen quality and sexual behavior by differential transcriptome analysis of the hypothalamus and pituitarium of male ducks. Genes including proopiomelanocortin (POMC), complexin 2 (CPLX2), hyaluronan and proteoglycan link protein 2 (HAPLN2), early growth response 4 (EGR4), tox high mobility group box family member 3 (TOX3), and muts homolog 4 (MSH4) were identified as key candidate genes affecting the HPG axis. Our findings provide new insights into the regulatory mechanisms underlying reproductive performance in male poultry and offer a reference for breeding programs aimed to improve reproductive performance in ducks.

Enhancing semen quality, antioxidant status and sex hormones of V-line rabbit bucks fed on supplemented diets with dried moringa leaves.

This study was conducted to evaluate the impact of dried moringa leaves (DML) on semen quality, immunoglobulin measurements, blood metabolic analysis, antioxidant status and sex hormones of V-line rabbit bucks. Twenty-four mature rabbit bucks of 7-8 months-old of proven fertility with an average weight (2.8 ± 0.4 kg) were classified into four treatments (basal diet with 0, 750, 1500 and 3000 mg DML/kg diet) and 6 bucks per treatment were used. Seminal plasma cholesterol (p = 0.013) and triglycerides (p = 0.0001) of all moringa supplemented rabbits were higher than the control. Moringa leaves enhanced (p < 0.05) serum testosterone, follicle stimulating hormone and luteinizing hormone. Total antioxidant capacity, superoxide oxidase, glutathione peroxidase and catalase in seminal plasma of moringa rabbits were higher (p < 0.05) than the control. Serum total lipid (p = 0.002), cholesterol (p = 0.008), triglycerides (p = 0.019) and blood urea (p < 0.05) were lower in DML rabbits. Moringa leaves improved total antioxidant capacity (p = 0.0001), glutathione peroxidase (p = 0.0001) and superoxide oxidase (p = 0.037) in rabbits versus control. Rabbits consuming DML had higher (p < 0.05) immunoglobulin IgG and IgM than control. Results suggest that DML improved rabbits' antioxidant, and immunological status, semen quality, and sex hormones, but DML (750 mg/kg diet) is recommended because it was more effective than the other two high doses.

Sexual behaviour response, testicular development and semen quality of New Zealand white rabbit bucks as influenced by dietary chitosan.

The present study was conducted to determine the effects of dietary chitosan supplementation on sexual behaviour responses, testicular development, and semen quality traits of New Zealand White (NZW) rabbit bucks. Twenty-four 5-week-old rabbit bucks were used in this experiment. Animals were grouped into four equal experimental groups: the control group was fed only on a basal diet, whereas the other groups were fed the basal diet supplemented with three levels of chitosan at 0.2, 0.4, or 0.6 g/kg, respectively. Also, bucks that received chitosan at 0.2 and 0.4 g/kg had a significantly earlier time of sexual libido (p ≤ .05) and had significantly higher ejaculate volume and sperm concentration than other groups (p ≤ .001). Furthermore, basic and sexual behaviours were significantly improved in bucks fed chitosan at 0.2 and 0.4 g/kg compared with other groups. Therefore, it could be concluded that using chitosan at 0.2 and 0.4 g/kg enhanced sexual behaviour, improved semen quality, and reproductive efficiency in the NZW rabbit bucks.

Maternal intake of folate and folic acid during pregnancy and markers of male fecundity: A population-based cohort study.

BACKGROUND: Poor male fecundity is of concern, and a prenatal origin has been proposed. Folate, a methyl donor involved in DNA methylation, is essential for normal fetal development by regulating gene expression during different periods of fetal development. Thus, prenatal exposure to low maternal folate intake might have a programing function of the developing reproductive organs. OBJECTIVES: To examine the association between maternal intake of folate from diet and folic acid from supplements during pregnancy and markers of fecundity in young men. MATERIALS AND METHODS: We conducted a follow-up study using a Danish mother-son cohort of 787 young men born 1998-2000. Percentage differences in semen characteristics, testes volume, and reproductive hormone levels were analyzed according to total folate calculated as dietary folate equivalents from diet and supplements in midpregnancy, using multivariable negative binomial regression models. Total folate was analyzed in quintiles, continuous per standard deviation decrease (SD: 318 µg/day) and as restricted cubic splines. RESULTS: Low maternal intake of total folate was associated with lower total sperm count (-5% (95% confidence intervals [CI]: -11%; 2%)), a lower proportion of non-progressive and immotile spermatozoa (-5% [95% CI: -8%; -3%]), and lower testes volume (-4% [95% CI: -6%; -2%]) per SD decrease in total folate intake. Spline plots supported these findings. DISCUSSION: The finding of a lower proportion of non-progressive and immotile spermatozoa, and hence a higher proportion of motile spermatozoa, in men of mothers with a lower intake of total folate in midpregnancy was surprising and may be a chance finding. CONCLUSION: Lower maternal intake of total folate in midpregnancy was associated with lower sperm count and lower testes volume, however, also with a lower proportion of non-progressive and immotile spermatozoa in adult men. Whether this actually affects the ability to obtain a pregnancy warrants further investigation.

The impact of varying doses of moringa leaf methanolic extract supplementation in the cryopreservation media on sperm quality, oxidants, and antioxidant capacity of frozen-thawed ram sperm.

To increase rams' post-thaw semen quality following cryopreservation, this study used enriched Tris-based diluent with varying amounts of moringa leaf methanolic extract (MLME). The antioxidant activity, total phenolic, and total flavonoid content were all assessed in MLME. The sperm of five healthy Awassi rams were collected, divided into 4 equal aliquots, and diluted [1:5; (v/v)] in Tris-citrate-glucose extender supplemented with 0.48, 0.56, and 0.64 mg MLME/ml or without MLME supplementation (control). The percentages of sperm total motility (STM, %), sperm progressive motility (SPM, %) and viability (V, %), abnormal morphology (AM, %), membrane functional integrity (MFI, %), and acrosome integrity (AI %) were measured. Malondialdehyde (MDA), nitric oxide (NO), ascorbic acid (AA), superoxide dismutase (SOD), glutathione peroxidase (GPx), total cholesterol (TC), low-density lipoproteins (LDL), lactate dehydrogenase (LDH), alkaline phosphatase (ALP), zinc (Zn), and copper (Cu) were measured. The total phenolic gallic acid and flavonoid catechin (equivalent) contents were 19.78 mg/g and 11.94 mg/g, respectively. 2,2-Diphenyl-1-picrylhydrazyl (34.37 mM TE/g) and 2,2'-azino-bis/3-ethylbenzothiazoline-6-sulfonic acid (53.47 mM TE/g) were found in MLME. MLME had a 64.59 mM TE/g ferric-reducing power. In comparison to control, the addition of 0.64 mg/ml MLME to Tris-based extender resulted in the highest (P < 0.001) STM (55.22 ± 0.98), SPM (45.41 ± .70), SV (60.01 ± 1.05), MFI (75.23 ± 0.77), and AI (73.13 ± 0.72) and the lowest (P < 0.001) AM (21.34 ± 0.72) values. In comparison to the control, the addition of 0.56 mg/ml semen extender resulted in lower STM, SPM, SV, MFI, and AI with higher AM percentages. MDA (P = 0.03), NO (P = 0.012), CHO (P = 0.0001), and LDL (P = 0.004) were reduced by 0.64 mg/ml MLME, while AA (P = 0.017) and SOD (P = 0.0001) were elevated. In conclusion, the highest copper (P = 0.006) and lowest zinc concentrations in MLME (0.48 mg/ml extender) deteriorated the post-thaw semen quality, prompting us to suggest the addition of 0.64 mg MLME to rams' Tris-based semen extender.

Longitudinal semen parameter assessments and live birth: variability and implications for treatment strategies.

OBJECTIVE: To examine whether semen parameters are associated with live birth among couples seeking infertility treatment after accounting for semen parameter variability. DESIGN: Folic Acid and Zinc Supplementation Trial (FAZST) prospective cohort. SETTING: Four US reproductive endocrinology and infertility care study centers, 2013-2017. PATIENT(S): Couples (n = 2,369) seeking fertility consultations at 4 US infertility care study centers. INTERVENTION(S): Semen volume, pH, sperm viability, morphology, progressive and total motility, concentration, count, and total and progressive motile count assessed at baseline and at 2, 4, and 6 months after enrollment. MAIN OUTCOME MEASURE(S): Log-binomial models stratified by fertility treatment received (in vitro fertilization [IVF], intrauterine insemination [IUI], ovulation induction [OI], or no treatment) estimated risk differences (RDs) between semen parameter quartiles and live birth and accounted for multiple semen assessments per person. We accounted for abstinence time, the biological interdependence of semen parameters, and potential selection bias because of loss to follow-up. RESULT(S): Among couples using OI only or no treatment, 39% had a live birth, and relative to the highest quartile, the lowest quartiles of morphology (RD, -19 [95% CI, -23 to -15] per 100 couples), motility (RD, -13 [95% CI, -17 to -9]), concentration (RD, -22 [95% CI, -26 to -19]), and total motile count (RD, -18 [95% CI, -22 to -14]) were associated with fewer live births. For IUI, 26% had a live birth, and the lowest quartiles of volume (RD, -6 [95% CI, -11 to -0.4]), concentration (RD, -6 [95% CI, -11 to -0.1]), count (RD, -10 [95% CI, -15 to -4]), and total motile count (RD, -7 [95% CI, -13 to -1]) were associated with fewer live births. For IVF, 61% had a live birth, and only morphology (Q1 RD, -7 [95% CI, -14 to 0.2]; Q2 RD, -10 [95% CI, -17 to -2.2]) was associated with live birth. CONCLUSION(S): Semen parameters are critical in couples undergoing OI/IUI. Only low morphology was important for live birth after IVF. Although data supporting the use of semen parameters are fragmented across differing populations, current findings are generalizable across the range of male fertility and couple fertility treatments, providing evidence about which semen parameters are most relevant in which settings. CLINICAL TRIAL REGISTRATION NUMBER: NCT#01857310.

L-carnitine improves quality parameters and epigenetic patterns of buck's frozen-thawed semen.

Buck sperm cryopreservation is an effective method to distribute qualified sperm for reproductive purposes, but this procedure reduces sperm quality. The current study was conducted to investigate the effect of L-carnitine (LC) on the quality and epigenetic patterns of buck's post-thawed semen. Semen samples were collected from five male goats twice a week and diluted in extenders supplemented with 0 (LC-0), 1 (LC-1), 5 (LC-5) and 10 (LC-10) mM LC. Samples were cryopreserved according to standard protocol. After thawing, motility characteristics, lipid peroxidation, membrane functionality, abnormal morphology, mitochondrial activity, acrosome integrity, epigenetic modifications, viability, apoptotic-like changes and DNA fragmentation were assessed. Samples supplemented with 5 mM LC showed greater (P ≤ 0.05) total motility, progressive motility, membrane functionality, mitochondrial activity, acrosome integrity, DNA methylation, viability, and lower (P ≤ 0.05) apoptotic-like changes. Lipid peroxidation was lower (P ≤ 0.05) in LC-5 and LC-10 compared to the control group. Addition of LC to the cryopreservation extender had no effect (P > 0.05) on velocity parameters, abnormal morphology, histone modifications, or DNA fragmentation. In conclusion, supplementing the cryopreservation extender with 5 mM LC significantly preserves the quality of buck sperm after the cryopreservation process.