Effects of organic and inorganic selenium supplementation on frozen-thawed ram semen at two cooling periods.

The aim of this study was to evaluate the effects of different selenium compounds on the sperm quality of cryopreserved ram semen. Ejaculates from four rams, collected using an artificial vagina heated to 38 °C, were individually evaluated. The approved ejaculates were pooled and diluted (1:1 v:v) in Tris-egg yolk extender (20%, v/v) and separated into two control groups, one cooled for 2 h and the other for 4 h. The pooled ejaculates at the two cooling periods were supplemented with two doses (0.5 and 1 µg/mL) of organic selenium (ORG), and inorganic selenium (SeNa), each. The samples were packed in 0.25 ml straws, at a concentration of 400 × 106 sperms/mL and stored in liquid nitrogen. The straws were thawed in a water bath at 37 °C for 20 s, and the samples were subjected to sperm kinetics evaluation by Computer Assisted Semen Analysis software. Sperm membrane integrity, acrosome morphology, and mitochondrial potential were assessed. In addition, oxidative stress markers reactive oxygen species (ROS), ferric reducing antioxidant power (FRAP), thiobarbituric acid reactive species (TBARS), and glutathione peroxidase (GPx) enzyme activity) were also evaluated. No significant improvement was observed in the ram semen quality at the two cooling times. Supplementation of the freezing extender with 0.5 µg/mL ORG, subjected to 4 h cooling period, increased the sperm motility when compared with the control group at the same cooling time. In addition, the 0.5 µg/mL SeNa group, under the 2 h cooling period, showed an increase in sperm motility when compared to the control group at the same cooling period. Considering the importance of sperm motility as a fertility parameter, our study indicates that supplementation with ORG and SeNa can help improve the total motility of the cryopreserved ram semen.

Treatment With Cholecalciferol Leads to Increase Of Selected Semen Parameters in Young Infertile Males: Results of a 6-month Interventional Study.

High incidence of infertility along with low vitamin D levels was detected in otherwise healthy young men. The aim is to observe the effect of vitamin D supplementation on semen parameters as assessed by semen analysis in infertile men. In total, 45 men (mean age 36.6 years) in consecutive order were included, of whom 34 finished the study. Subjects were supplemented by vitamin D (cholecalciferol) 2500 IU/day. Vitamin D levels were assessed by HPLC. Semen analysis was performed strictly following 2010 WHO guidelines. Study periods were baseline and month 6. During follow-up, 20 %, 7.4 %, 22 % and 0.7 % increase in serum vitamin D levels, progressive sperm motility, sperm concentration and sperm morphology, respectively, were observed (all p<0.05). At follow-up end, 9 patients (26 %) reached normal sperm parameters of whom 2 fertilized their partner. There was no correlation between vitamin D and semen parameters observed. This study proves that vitamin D supplementation is possibly a modulator of sperm parameters in vitamin D deficient, otherwise healthy men. Although a direct relationship between vitamin D and sperm parameters was not observed obtaining adequate vitamin D levels could likely play a role in the male factor of infertility.

Dietary ß-glucan influences the expression of testicular aquaporins, antioxidative defence genes and sperm quality traits in endangered golden mahseer, Tor putitora (Hamilton, 1822).

The effect of dietary ß-glucan on seminal plasma composition, sperm characteristics, expression of aquaporins, and antioxidative defence genes of golden mahseer was evaluated. For that, four experimental diets containing 0 (control), 0.5, 1, and 1.5% ß-glucan were fed to male golden mahseer brooders for 130 days. Feeding of 0.5% ß-glucan was found to improve sperm characteristics, viz. sperm count, motility, viability, and morphology with no effect on gonadosomatic index and seminal plasma energy resources. The marked down-regulation in the transcript abundance of testicular aqp3a noticed in 1.5% ß-glucan fed brooders corresponds to their poor sperm quality. Further, the mRNA expression of genes encoding antioxidant enzymes, namely gst and sod1, was lowest in 0.5% ß-glucan fed brooders. In contrast, control and higher ß-glucan (1 and 1.5%) groups displayed relatively higher expression levels of testicular gst and sod1. On the other hand, the higher seminal plasma total antioxidant capacity observed in 0.5 and 1% ß-glucan fed brooders indicated increased scavenging ability of reactive oxygen species. Overall, supplementation of 0.5% ß-glucan improved sperm quality and antioxidative potential, but the higher inclusion (1.5%) negatively affected sperm characteristics. Collectively, dietary ß-glucan (0.5%) can be a practical approach to developing quality broodstock of golden mahseer.

The Effect of Oral Vitamin E on Semen Parameters and IVF Outcome: A Double-Blinded Randomized Placebo-Controlled Clinical Trial.

BACKGROUND: Male infertility is a main clinical problem that affects about 7% of all men worldwide. Many patients with male infertility are caused by a reduced antioxidant capacity of semen. Several antioxidant supplements, especially vitamin E, are proposed to help male infertility treatment. This project was goaled to study the effects of oral synthetic vitamin E (400 IU/day) for eight weeks on betterment of semen parameters and pregnancy rate. METHODS: After dropping the cases, 124 infertile couples with a male factor who were admitted to the IVF program were included. The male patients with idiopathic abnormal motility and/or morphology were randomized into two groups: 61 receiving vitamin E and 63 as the control group receiving placebo for eight weeks. The pretreatment semen parameters of both groups were compared with those of posttreatment. The pregnancy outcomes were considered between the two groups. RESULTS: There were no significant differences statistically between before and after treatment in the term of sperm volume, count, motility, and morphology. Furthermore, the IVF outcomes of the two groups were not different significantly, either. Interestingly, the percent of normal sperm in the placebo group was significantly decreased after eight weeks. CONCLUSION: Vitamin E supplementation might neutralize free radical activity to keep sperm from more oxidative damages. Further studies regarding the influence of higher acceptable doses of vitamin E on semen characteristics and fertility rates are needed. This study was registered as a two-arm, blinded, randomized, placebo-controlled clinical trial (IRCTID: IRCT2014020616506N1, 2014-03-18).

Soursop juice enhanced seminal antioxidant defence and semen quality of rabbit bucks in extremely dry climatic condition of Southwestern Nigeria.

Heat induced stress associated with dry climatic conditions in the tropics does have adverse effects reproduction in rabbits, and this in-turn impacts negatively on the income of rabbit farmers. However, natural products might prove to be a reliable, safe and cheap remedy for ameliorating reproductive such anomalies in rabbits. The potential of soursop for mitigating heat induced reproductive deficiency in rabbit bucks was investigated during the peak of dry climatic conditions in southwestern Nigeria. Sixty mixed breed (New Zealand white x Chinchilla) adult rabbit bucks were allotted to five treatments of four replicates (3 bucks per replicate) each in a completely randomized design. Soursop (Annona muricata) fruit was processed into juice using standard procedures and was designated as soursop juice. The juice was administered orally daily per kg body weight (BW), 0.55ml/kgBW distilled water (control), 0.55 ml/kgBW soursop juice, 1.11 ml/kgBW soursop juice, 1.67 ml/kgBW soursop juice and 2.22 ml/kgBW soursop juice to designated treatments 1 to 5, respectively for 56 days. Semen samples were collected with an improvised artificial vagina on the 28th and 56th day of the study, semen quality and seminal oxidative status were evaluated using standard procedures. Results showed that rabbit buck exposed to heat stress had lower semen quality, seminal antioxidants and increased seminal lipid peroxidation. However, the consumption of soursop juice lowered lipid peroxidation and enhanced (p < 0.05) antioxidant production in the seminal fluid of heat-stressed bucks than bucks on control group. Bucks' semen quality and antioxidant status peaks in heat-stressed bucks gavaged 2.22 ml/kgBW soursop juice and gives 100% recovery from the effects of heat induced stress. It can be concluded that 2.22 ml/kgBW soursop juice administered for 56 days did enhance spermatozoa quality and mitigated lipid peroxidation by improving antioxidant capacity of male rabbits in a dose dependent manner on extremely dry climatic conditions.

Effects of hydroethanolic extracts of Terminalia chebula and Thymbra spicata on ram fresh semen under normal and oxidative stress conditions.

The objective of this study was to evaluate protective effects of hydroethanolic extracts of Terminalia chebula and Thymbra spicata on viability, lipid peroxidation (LPO) and DNA integrity of ram fresh semen under normal and oxidative stress (OS) conditions. Antioxidant activities of different concentrations of Terminalia chebula and Thymbra spicata extracts were evaluated with DPPH assay. Semen samples were taken from three fertile adult rams. After diluting semen with Tris-base extender, different concentrations of Terminalia chebula and Thymbra spicata (30, 300, and 3000 µg/ml) extracts were used under normal and induced OS conditions. The group not receiving any supplements was considered as control group. A total of 50 µM hydrogen peroxide was used to induce OS. MTT solution was added to each of treatment groups which were kept in an incubator at 37°C for 2 h. After incubation, readings were obtained by ELISA reader. DNA integrity and LPO were determined with acridine orange (AO) staining and malondialdehyde (MDA) assay. Higher concentrations of Terminalia chebula and Thymbra spicata extracts preserved viability and DNA integrity while reducing MDA concentrations compared to other treatment groups. Also, under induced OS, higher concentrations of both extracts reduced detrimental effects of H2 O2 . In conclusion, it seems that addition of Terminalia chebula and Thymbra spicata extracts can reduce induced OS in spermatozoa.

The effects of Vitamin D3 supplementation on Spermatogram and endocrine factors in asthenozoospermia infertile men: a randomized, triple blind, placebo-controlled clinical trial.

OBJECTIVE: Evaluate the effects of vitamin D3 (VD3) on sperm parameters and endocrine markers in infertile men with asthenozoospermia. MATERIALS AND METHODS: This randomized, triple-masking, placebo-controlled clinical trial conducted on 86 asthenozoospermia infertile men with serum 25 hydroxy vitamin D3 (25(OH)VD3) < 30 ng/ml in the infertility clinic of Ahvaz Jahad daneshgahi, Iran. Patients were randomly allocated to groups A and B, who received daily 4000 IU VD3 and matching placebo respectively for 3 months. Demographic data, dietary intake, physical activity, sun exposure, anthropometric indices, serum 25(OH)VD3, luteinizing hormone (LH), follicle-stimulating hormone (FSH), total testosterone (T), estradiol (E2),, sex hormone-binding globulin (SHBG), free androgen index (FAI = T/SHBG. 100), T/LH and T/E2 ratios, prolactin (PRO), parathyroid hormone (PTH), osteocalcin (OCN), phosphorus and sperm parameters were assessed. RESULTS: Three months VD3 supplementation with 4000 IU/day had no significant effects body weight, body mass index (BMI), waist circumference (WC), body fat (BF), serum, OCN, LH, FSH, T, E2, SHBG, PRO, T/E2 ratio, FAI, semen volume, sperm count and normal sperm morphology. It increases serum 25(OH)VD3, PTH and phosphorus and seminal and serum calcium, T/LH ratio and total and progressive sperm motility and decreased significantly compared to the baseline and placebo group. CONCLUSION: VD3 supplementation may affect sperm motility in men with asthenozoospermia and serum 25(OH)VD3 < 30 ng/ml. TRIAL REGISTRATION: Iran Clinical Trials Registry, ID: IRCT20151128025274N4, registered on 28 March 2018, URL of trial registry record: https://www.irct.ir/trial/29983.

IGF-1 supplementation in semen affects mitochondrial functional and calcium status of buffalo sperm following cryopreservation.

This study was designed to examine the effects of seminal insulin-like growth factor-1 (IGF-1) supplementation on structural and functional properties of buffalo sperm post cryopreservation. Semen ejaculates from buffalo bulls (n = 6) were proportioned into four aliquots and diluted with egg yolk-based extender. Prior to equilibration, IGF-1 was added to extender as four treatments: group IGF0 (no supplementation), IGF150 (150 ng/mL), IGF250 (250 ng/mL) and IGF350 (350 ng/mL). The extended semen was transferred into 0.25 mL mini-straws, equilibrated (4 °C at 4 h), and cryopreserved. Total sperm motility was greater (P < 0.05) when there was the IGF150 treatment compared with values for other groups. Furthermore, with the IGF150 treatment there was the least and greatest (P < 0.05) mitochondrial superoxide status and membrane potential, respectively. Similarly, with the IGF150 treatment there was a greater (P < 0.05) sperm membrane integrity with a lesser (P < 0.05) calcium status compared to values for the other groups. In conclusion, seminal IGF-1 supplementation affects the structural and functional properties of buffalo sperm following cryopreservation.

Improving the quality and in vitro fertilization rate of frozen-thawed semen of buffalo (Bubalus bubalis) bulls with the inclusion of vitamin B12 in the cryopreservation medium.

This study was designed to evaluate the effects of vitamin B12 in cryopreservation medium on frozen-thawed semen of buffalo (Bubalus Bubalis) bulls. Semen from five bulls (fertility-proven) were diluted in five aliquots not supplemented (control), or supplemented with 1, 2, 4, or 5 mg/mL of vitamin B12 and evaluated using the Computer Assisted Sperm motion Analysis, antioxidant enzymes, lipid peroxidation (LPO), reactive oxygen species (ROS), ATP concentrations, and in vitro fertilization rate (%). Sperm progressive motility, rapid velocity (%), mitochondrial potential, and acrosome integrity were greater (P < 0.05) with supplementation of 4, and 5 mg/mL vitamin B12 than the control sample. Similarly, compared with the control, samples with 5 mg/mL vitamin B12 supplementation had markedly greater average-path, straight-line, and curved-line velocities (µm/sec). Semen samples supplemented with 2, 4 and 5 mg/mL vitamin B12 had greater concentrations of GPx (U/mL) and SOD (U/mL), whereas LPO (µM/mL) was less (P < 0.05) compared with the control sample. Seminal plasma ROS concentrations (104/25 × 106) were less in the 5 mg/mL vitamin B12 supplemented than control sample. Semen samples supplemented with 5 mg/mL of vitamin B12 had greater concentrations of ATP than control and the 1 mg/mL vitamin B12 supplemented sample. Semen samples supplemented with 5 mg/mL of vitamin B12 had greater plasmalemma and DNA integrities (%) than the control sample. In summary, vitamin B12 supplementation augments semen quality, as evidenced by values for CASA variables, antioxidant enzymes, and ATP concentrations, which may occur as a consequence of inhibition in LPO and ROS production by buffalo spermatozoa.

Supplementation of the BIOXcell extender with the antioxidants crocin, curcumin and GSH for freezing bull semen.

Semen cryopreservation is routine in cattle, but the results of artificial insemination need improvement. A strategy to these aims is the supplementation of the freezing extender with novel antioxidants. This study aimed at testing the natural antioxidants curcumin and crocin as supplements to the commercial extender BIOXcell for freezing semen from 8 Holstein bulls. We tested curcumin at 0.05 and 0.1 mM (CU0.05, CU0.1) and crocin at 0.5 and 1.5 mM (CR0.5, CR1.5), with 0.5 mM reduced glutathione (GSH0.5) as reference, and a control (CTL, without supplementation). The samples were evaluated post-thawing and after 5 h at 38 °C by CASA for motility and flow cytometry for viability, apoptotic, capacitation, acrosomal status, cytoplasmic and mitochondrial reactive oxygen species (ROS) production, and chromatin status (SCSA). Control and GSH0.5 showed similar results, possibly because of the good protection from BIOXcell. CU0.05 and CU0.1 showed little effects but increased cytoplasmic ROS production and motility ALH. CR0.5 and CR1.5 decreased viability and increased apoptotic features significantly post-thawing and after the incubation, resulting in lower motility (significant after the incubation) but decreasing SCSA %HDS (loose chromatin). Whereas crocin at these concentrations seems incompatible with BIOXcell, maybe because of a prooxidant activity, curcumin use merits further research, considering the elevation of ROS with no significant negative effects.

Exposure to multiple metals/metalloids and human semen quality: A cross-sectional study.

BACKGROUND: Exposure to metals/metalloids, including essential and nonessential elements, has been associated to male reproductive health in animals. However, findings from human studies are inconsistent. OBJECTIVES: To investigate the impact of exposure to multiple metals/metalloids at environmental levels on the conventional human semen-quality parameters. MATERIALS AND METHODS: Men living in rural or industrial areas were recruited by personalized letters. No exclusion criteria were applied. Each man provided one semen sample and one blood sample. We analyzed the semen sample both to determine conventional sperm parameters (concentration, progressive motility and normal forms) and to quantify lead (Pb), cadmium (Cd), mercury (Hg), arsenic (As), nickel (Ni), vanadium (V) and selenium (Se) levels. The levels of these metals/metalloids were also quantified in venous blood and spermatozoa samples. Associations between the blood/seminal plasma metal/metalloid levels and semen quality parameters were assessed using confounder adjusted logistic regression models. Correlation and interactions between blood/seminal plasma and semen metal/metalloid levels were investigated using the Spearman's correlation. RESULTS: We found a positive association of seminal plasma cadmium level with lower Total count (OR = 4.48, 95%CI 0.25-80); whereas lead (OR = 4.51, 95%CI 0.86-23) and cadmium (OR = 3.45, 95%CI 0.77-16) seminal plasma levels had a positive association with progressive sperm motility. Overall, these associations remained suggestive after adjustment, though statistically unstable risks. Finally, we found weak interactions between beneficial effects of Se and detrimental ones only for Cd and Pb blood level on sperm concentration, total sperm count and progressive sperm motility. CONCLUSIONS: Our findings suggest that environmental exposure to Pb and Cd contributes to a decline in human semen quality, whereas Se can have beneficial effects. Measurements of metals/metalloids in the seminal fluid may be more predictable of semen quality than conventional blood measurements.

Maternal dietary supplementation with grape seed extract in reproductive hens increases fertility in females but decreases semen quality in males of the F1 generation.

Genetic selection in parental broiler breeders has increased their susceptibility to metabolic disorders and reproductive dysfunction. We have recently shown that maternal dietary grape seed extract (GSE) supplementation in hens improves fertility parameters, egg quality, oxidative stress in different tissues and the quality of F1 chicks. Here, we analysed the growth and fertility (both female and male) of the F1 generation animals and the quality of their offspring (F2 generation). Eggs issued from hens supplemented with GSE presented lower ROS production than control hens, suggesting a change in the embryonic environment. However, this did not affect the growth nor the body composition of male and female F1s from hatching to adulthood (37 weeks of age). At 37 weeks of age, the biochemistry analysis of the GSE-F1 muscle has revealed an increase in sensitivity to oxidative stress and a slight change in lipid composition. Both male and female F1-GSE groups presented a delay in puberty with a lower testis volume at 30 weeks of age and lower ovary development at 26 weeks of age. Adult GSE-F1 males did not present histological alterations of seminiferous tubules or semen production, but the semen quality was degraded due to higher oxidative stress and DNA-damaged spermatozoa compared with control F1 animals. In adult GSE-F1 females, despite the delay in puberty, the females laid more eggs of better quality (fewer broken eggs and a higher hatching rate). At hatching, the weight of the chicks from GSE-F1 females was reduced, and this effect was stronger in F2 male chicks (F2) compared with F2 control chicks (F2), because of the lower muscle volume. In conclusion, we can raise the hypothesis that maternal dietary GSE supplementation produces eggs with change in embryonic metabolism, which may affect in adulthood the fertility. The data obtained from the F1-GSE group pointed to a sex-specific modification with higher egg quality in females but semen sensitive to stress in males. Finally, male F2 chicks were leaner than control chicks. Thus, maternal dietary grape seed extract (GSE) supplementation in hens may impact on the fertility of the offspring in a sex-specific manner in subsequent generations.

Efecto de la suplementación del medio de incubación con extracto de Berberis microphylla (calafate) sobre la funcionalidad espermática en semen bovino descongelado / Effect of supplementation of incubation medium with Berberis microphylla (calafate) extract on sperm functionality in thawed bovine semen

RESUMEN: En el semen criopreservado, los procesos de congelación/descongelación y posterior manipulación, dañan las células espermáticas provocando disminución de la capacidad fecundante de los espermatozoides descongelados. Estos procesos han sido asociados con el estado de estrés oxidativo (EO) inducido por altos niveles de especies reactivas de oxígeno (EROS), causando daño a la función y estructura espermática. Los espermatozoides descongelados pueden ser protegidos de este daño, con la adición de antioxidantes (AO) al medio de incubación. El fruto de Calafate (Berberis microphylla G. Forst.) posee una alta capacidad antioxidante, lo que hace interesante investigar el efecto de sus componentes antioxidantes en estos procesos biotecnológicos especialmente postdescongelación. El objetivo de este estudio fue determinar el efecto de la suplementación de extracto liofilizado de fruto de Calafate (ELC), sobre la calidad espermática post-descongelación. Previamente se caracterizó el ELC, determinando la actividad antioxidante y metabolitos como fenoles y antocianinas; posteriormente, espermatozoides de bovino descongelados fueron incubados en un medio base suplementado con diferentes concentraciones de ELC. Post-incubación se evaluó la motilidad progresiva; la viabilidad e integridad de la membrana plasmática (SYBR14- PI) y acrosomal (FITC-PNA/PI) y la peroxidación lipídica (BODIPY) por citometría de flujo. La caracterización de ELC demostró que tanto la actividad antioxidante como los fenoles y antocianinas incrementan concomitante con el aumento de la concentración de ELC. La adición de ELC al medio de incubación, dependiendo de la concentración y tiempo de incubación, sería eficaz en proteger la motilidad, viabilidad e integridad de la membrana plasmática y disminuir la lipoperoxidación en los espermatozoides de bovino descongelados.

SUMMARY: In cryopreserved semen, the freezing/thawing process following of manipulation, damage the sperm cell, decreasing the fertilizing capacity of the thawed sperm; being one of the main factors of this damage the oxidative stress. The sperm once thawed can be protected from this damage, with the addition of antioxidants to the incubation medium. The Calafate fruit (Berberis microphylla G. Forst.) has a high antioxidant capacity, making it an interesting resource for investigating the effect of its antioxidant components on biotechnological processes. The objective of this study was to determine the effect of supplementation of Calafate fruit lyophilized extract (ELC) on sperm quality. The lyophilized extract of the Calafate fruit was characterized, determining the antioxidant activity and metabolites such as phenols and anthocyanins; subsequently, thawed bovine sperm were incubated in a medium supplemented with different concentrations of ELC. Post-incubation, progressive motility was evaluated. By flow cytometry, the viability and integrity of the plasma (SYBR14-PI), and acrosomal (FITC-PNA / PI), as well as lipid peroxidation (BODIPY), was determined. The characterization of Calafate fruits lyophilized extract indicated that antioxidant activity, phenols and anthocyanins increased concomitantly with the increase of dose extract used. The addition of ELC to the incubation medium, depending on the concentration and incubation time, would be effective to protect motility, viability and integrity of the plasma membrane and decreased lipid peroxidation in thawed bovine sperm.

Effect of adding different concentrations of L-arginine to Tris-yolk extender on the quality of sub-fertile ejaculates in buffalo.

To investigate the effect of supplementation of L-arginine (AR) on sub-fertile buffalo-bulls' ejaculates, 25 ejaculates of poor motility (40 to 55%) were collected by artificial vagina from 5 buffalo-bulls and extended with Tris-yolk extender (1:10) supplemented with different concentrations of AR (0, 3, 4, 5, and 6 mM). Semen was cooled gradually to 4 °C within 2 h and incubated at 4 °C for additional 2 h. Incubated semen samples were evaluated by computer-assisted semen analysis. Results showed that addition of 5 mM AR increased (P < 0.05) total sperm motility and rapid progressive motility percentages, while decreased (P < 0.05) non-motile sperm and static sperm percentages compared with AR-free (control) extender. Increasing the AR level to 6 mM increased (P < 0.05) the percentages of sperm progressive motility and rapid and slow progressive motilities, while decreased (P < 0.05) the non-progressive sperm motility percentages compared with AR-free extender. Supplementation of 5 mM AR improved (P < 0.05) sperm straight linear, curve linear, and average path velocities (36 ± 0.13, 20.6 ± 5.3, and 33.2 ± 8.5, respectively) in comparing with control and other AR treatments. Addition of AR (5 and 6 mM) improved (P < 0.05) the percentages of vitality (89.8 ± 1.9 and 80.0 ± 3.4, respectively), normality (44.3 ± 3.6 and 44.8 ± 1.5, respectively), and functional sperm (20.4 ± 8.6 and 21.0 ± 0.61, respectively), and decreased abnormal neck and tail percentages compared with AR-free extender. All AR levels decreased (P < 0.05) the abnormal neck and tail percentages. Addition of all AR levels had no significant (P > 0.05) effect on the activity of aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase in semen extender. Supplementation of Tris-yolk extender with L-arginine (5 or 6 mM) can improve sperm motility, velocity, vitality, and functional sperm and can decrease tail and neck abnormalities of sub-fertile buffalo ejaculate after 4 h incubation at cool temperature.

The role of alpha lipoic acid in female and male infertility: a systematic review.

OBJECTIVE: Infertility is an increasingly frequent health condition, which may depend on female or male factors. Oxidative stress (OS), resulting from a disrupted balance between reactive oxygen species (ROS) and protective antioxidants, affects the reproductive lifespan of men and women. In this review, we examine if alpha lipoic acid (ALA), among the oral supplements currently in use, has an evidence-based beneficial role in the context of female and male infertility. METHODS: We performed a search from English literature using PubMed database with the following keywords: 'female infertility', 'male infertility', 'semen', 'sperm', 'sub-fertile man', 'alpha-lipoic acid', ' alpha lipoic acid', 'lipoid acid', 'endometriosis', 'chronic pelvic pain', 'follicular fluid' and 'oocytes'. We included clinical trials, multicentric studies and reviews. The total number of references found after automatically and manually excluding duplicates was 180. After primary and secondary screening, 28 articles were selected. RESULTS: The available literature demonstrates the positive effects of ALA in multiple processes from oocyte maturation (0.87 ± 0.9% of oocyte in MII vs 0.81 ± 3.9%; p < .05) to fertilization, embryo development (57.7% vs 75.7% grade 1 embryo; p < .05) and reproductive outcomes. Its regular administration both in sub-fertile women and men shows to reduce pelvic pain in endometriosis (p < .05), regularize menstrual flow and metabolic disorders (p < .01) and improve sperm quality (p < .001). CONCLUSIONS: ALA represents a promising new molecule in the field of couple infertility. More clinical studies are needed in order to enhance its use in clinical practice.

Does alpha-lipoic acid-loaded nanostructured lipid carriers improve post-thawed sperm quality and ameliorate apoptosis-related genes of rooster sperm?

Oxidative stress could be prevented by antioxidant-loaded nanoparticles. The purpose of this study was to assess the effects of 10 (A10), 20 (A20), 30 (A30), 40 (A40), and 50 (A50) µM alpha-lipoic acid and alpha-lipoic acid nanostructured lipid carriers (ALN) at 10 (ALN10), 20 (ALN20), 30 (ALN30), 40 (ALN40), and 50 (ALN50) µM on post-thawed sperm quality, fertility, and apoptosis-related genes of rooster sperm. The extender supplemented with ALN30 led to higher total and progressive motility, straight-line velocity, and linearity in comparison to the control group. The ALN30 resulted in higher percentage of mitochondria activity and glutathione peroxidase level compared with control (P < 0.05). The extender supplemented with ALN30 led to lower percentage of apoptotic sperm, when compared with the control. CASPASE 3 expression in ALN30 was lower (P < 0.05) than the other groups. The results showed that BCL-2 mRNA expression of sperm was significantly (P < 0.05) higher in ALN30 compared with the other groups (P < 0.05). Higher percentages of fertility and hatchability rates were observed in ALN30 group. The results indicate that ALN30 could be regarded as a novel potential cryoprotectant for the cryopreservation of rooster semen. Therefore, nanostructured lipid carriers improve not only the active compound (such as alpha-lipoic acid) of biomedical applicability but also the potential for industrial application in sperm cryopreservation.

Effect of crocin and naringenin supplementation in cryopreservation medium on post-thaw rooster sperm quality and expression of apoptosis associated genes.

The aim of our study was to examine the effects of crocin (0.5 (C0.5), 1 (C1) and 1.5 (C1.5) mM) and naringenin (50 (N50), 100 (N100) and 150 (N150) µM) in cryopreservation extender for freezing rooster semen. Sperm motility, viability, abnormalities, membrane functionality, active mitochondria, apoptosis status, lipid peroxidation (LP), GPX, SOD, TAC, the mRNA expression of pro-apoptotic (CASPASE 3) and anti-apoptotic (Bcl-2) genes, fertile eggs, hatched eggs and hatching rate were investigated following freeze-thawing. C1 and N100 resulted in higher (P < 0.05) total motility and progressive motility in comparison to the control group. The C1 and N100 groups improved viability, membrane functionality and reduced lipid peroxidation. We found higher values for active mitochondria with C1 and N100 compared to control group. The C1 and N100 groups showed lower percentages of early apoptosis when compared with control group. Also, C1 and N100 had higher TAC, compared to the control group. The mRNA expressions of BCL-2 in the C1 and N100 groups were significantly higher than that of other treatments. The expression of CASPASES 3 was significantly reduced in C1 and N100 group (P < 0.05) when compared to control group. Significantly higher percentages of fertile eggs, hatched eggs and hatching rate were observed in C1 and N100 compared to the control group. In conclusion, crocin at 1 mM and naringenin at 100 µM seem to improve the post-thawing rooster semen quality, fertility and could protect the sperm by reducing the pro-apoptotic (CASPASE 3) and increasing anti-apoptotic (Bcl-2) genes.

Evaluation of the effect of vitamin D3 supplementation on quantitative and qualitative parameters of spermograms and hormones in infertile men: A Randomized controlled trial.

BACKGROUND: 25-Hydroxy Vitamin D3 is known to have an effect on reproductive system in both genders and may change the semen parameters in men. OBJECTIVE: Our study aimed to evaluate the effect of oral vitamin D3 supplementation on spermogram quantitative and qualitative parameters in infertile men. MATERIALS AND METHODS: This study was a triple-blind randomized controlled trial involving 62 infertile men with impaired spermatogonial tests. They were randomly divided into placebo and D3-supplemented groups. Spermograms and tests for LH (Luteinizing Hormone), FSH (Follicle Stimulating Hormone), TT (Total Testosterone), FT (Free Testosterone), SHBG (Sex Hormone Bonding Globulin), FAI (Free Androgen Index) and vitamin D3 levels were performed before and after the intervention. RESULTS: There were no significant differences between the two groups in parameters of the spermograms or serum levels of LH, FSH, TT, and FAI. In the intervention group, SHBG was significantly decreased after intervention (p = 0.01) and there was a significant increase in FT in the placebo group (p = 0.03). CONCLUSION: The intake of vitamin D3 did not change the quality and quantity of spermograms and serum levels of LH, FSH, TT, and FAI but affected FT and SHBG. Further studies are still needed to clarify the biological role of vitamin D3 on fertility particularly on male fertility. This study lays a foundation for more extensive studies on male infertility.

Escherichia coli membrane-derived oxygen-reducing enzyme system (Oxyrase) protects bubaline spermatozoa during cryopreservation.

The objective of this study was to determine the effectiveness of deoxygenation of semen extender using Escherichia coli membrane-derived oxygen scavenger (Oxyrase) on post-thaw quality of buffalo (Bubalus bubalis) spermatozoa. Sixteen semen ejaculates, four each from four bulls, were each divided into five equal fractions, diluted using Tris-egg yolk extender supplemented with different concentrations of Oxyrase (0, 0.3, 0.6, 0.9, and 1.2 U/ml), designated as treatments T1, T2, T3, T4, and T5, respectively, and cryopreserved. Immediately after thawing, Oxyrase did not improve sperm kinetics and motility; however, it improved the keeping quality (significantly lower deterioration of post-thaw sperm motility after incubation for 120 min) in T3. Further, T3 reduced (p < .05) cholesterol efflux and protected the intactness of the sperm plasma membrane. Flow cytometry with Fluo-3 AM/propidium iodide (PI) dual staining revealed the highest (p < .05) proportion of live spermatozoa with low intracellular calcium in T3. Oxyrase supplementation protected spermatozoa from premature capacitation which was confirmed by low expression of tyrosine-phosphorylated proteins (32, 75, and 80 kDa) and a relatively lower percentage of F-pattern (uncapacitated spermatozoa) in chlortetracycline assay. Importantly, the Oxyrase fortification decreased superoxide anion in a dose-dependent manner indicating reduced availability of oxygen at sperm mitochondrial level. Similarly, in Oxyrase-fortified sperm, malondialdehyde concentration, an index of lipid peroxidation, is also reduced in a dose-dependent manner. In conclusion, we demonstrate that deoxygenation of buffalo semen by Oxyrase has the potential of improving post-thaw sperm quality by overcoming the problem of cryocapacitation and oxidative damage during cryopreservation process.

Efecto de la borra de café sobre la biológica espermática: aproximación in vitro / Effect of spent coffee grounds on sperm biology: in vitro approach

OBJETIVO: El objetivo de este trabajo fue establecer el efecto de la borra de café sobre la movilidad y los parámetros funcionales de los espermatozoides humanos in vitro. MATERIALES Y MÉTODOS: La borra de café, un subproducto obtenido en establecimientos especializados en la preparación de café soluble a base de grano, se diluyo en tampón fosfato salino y se mezcló en proporciones iguales con las muestras de semen de 16 voluntarios aparentemente sanos. A cada muestra se le determinó el efecto sobre la movilidad espermática en función del tiempo (30, 60, 90 y 120 minutos, n=16) y sobre los parámetros funcionales (n=6) por medio de citometría de flujo: potencial de membrana mitocondrial, producción de especies reactivas de oxígeno y lipoperoxidación de la membrana espermática. RESULTADOS: La incubación de los espermatozoides con la borra de café evidencio un cambio positivo en la movilidad espermática. Adicionalmente, la incubación con la borra de café incremento significativamente el potencial de membrana mitocondrial en los espermatozoides. CONCLUSIÓN: La borra de café, seguramente debido a los compuestos antioxidantes, afecta positivamente la movilidad espermática aumentando el potencial de membrana mitocondrial. Por lo tanto, esto es un paso inicial en la búsqueda de un suplemento de origen natural que aumente la calidad seminal.

OBJECTIVE: The objective of this work is to establish the effect of spent coffee grounds on the motility and functional parameters of human spermatozoa, in vitro. MATERIALS AND METHODS: Spent coffee grounds, a by-product obtained in specialized establishments in the preparation of soluble coffee based on grain, was diluted in saline phosphate buffer and mixed in equal proportions with semen samples from 16 apparently healthy volunteers. Each sample was determined the effect on sperm motility as a function of time (30, 60, 90 and 120 minutes, n=16) and on functional parameters (n=6) by means of flow cytometry: mitochondrial membrane potential, reactive oxygen species production and membrane lipoperoxidation. RESULTS: The incubation of the spermatozoa with the spent coffee grounds showed a positive change in sperm motility. Additionally, incubation with spent coffee grounds significantly increased the mitochondrial membrane potential in human sperm cells. CONCLUSION: Spent coffee grounds, probably due to antioxidant compounds, positively affects sperm motility by increasing mitochondrial membrane potential. Therefore, this is an initial step in the search for a supplement of natural origin that increases seminal quality.