Plasma Membrane Functional Integrity Research Articles

Oxyrase-Mediated Improvement in the Quality and Fertility of Crossbred Boar Spermatozoa During Liquid Storage.

The present experiment was carried out to investigate the role of Oxyrase in preserving the invitro quality, redox status and invivo fertility of crossbred boar spermatozoa. A total of 24 ejaculates from 6 crossbred (n = 4 from each boar) boars were collected and extended in Beltsville Thawing Solution (BTS) in 1:2 ratio and divided into three aliquots. The first aliquot served as a control (without Oxyrase). Rest of the two aliquots were supplemented with 0.125 (T1) and 0.25 IU/mL Oxyrase (T2). Semen samples were preserved at 15°C for 5 days and kinematics of spermatozoa by CASA, semen quality parameters and oxidative stress status were evaluated at 0, 72 and 120 h of storage. The findings of studies revealed that supplementation of Oxyrase at 0.25 IU/mL resulted in higher (p < 0.05) total motility, progressive motility, plasma membrane integrity, acrosome integrity and functional integrity of plasma membrane at 72 and 120 h in comparison to the control group. Mitochondrial membrane potential (MMP) was higher (p < 0.05) at 72 and 120 h, whereas higher (p < 0.05) DNA integrity was observed at 120 h in T2. The lipid peroxidation (LPO) was lower (p < 0.05) and superoxide dismutase activity (SOD) and total antioxidant capacity (TAC) were higher (p < 0.05) in the T2 group at 120 h as compared to control. Invivo fertility trials indicated a higher (p < 0.05) litter size in T2 in comparison to other groups. The study concluded that the inclusion of Oxyrase at 0.25 IU/mL in the extender protects the crossbred boar spermatozoa against oxidative damage and improves the invivo fertility.

Effects of short-term oral letrozole on fresh semen parameters, endocrine balance, and prostate gland dimensions in domestic dogs

BackgroundAromatase 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).ResultsOur 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.ConclusionsAccording 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.

The effects of supplementation of vitamin D to the egg-yolk extender on cryopreservation of ram semen.

This study aimed to examine the effects of supplementation of vitamin D to the egg-yolk extender on characteristics of frozen-thawed ram semen. Semen samples obtained from adult rams were pooled and divided into five equal volumes. It was reconstituted with extenders containing different concentrations of vitamin D: 0 (control), 12.5 (VITD 12.5), 25 (VITD 25), 50 (VITD 50), and 100ng/mL (VITD 100), and then they were frozen. Sperm motility parameters, plasma membrane functional integrity, acrosomal integrity, DNA fragmentation, and mitochondrial membrane potential of the groups were evaluated after sperm thawing. Total motility and progressive motility were higher in VITD 50 than in all other groups (p<0.05). Higher sperm straightness, linearity, and wooble were higher in VITD 50 than in the control group (p<0.05). A similar pattern of VITD 50 was observed for plasma membrane integrity and mitochondrial membrane potential (p>0.05). In the study, it was observed that adding vitamin D to the extender had a beneficial effect on ram spermatological parameters. In addition, it was concluded that the use of the 50ng/mL vitamin D in the extender provided more effective protection than the other doses.

Insulin enhances drone semen quality after cryopreservation

The main objective of this study was to investigate the effect of insulin on drone semen quality after cryopreservation. Semen samples were obtained from sexually mature drones. The semen samples were pooled and diluted with varying concentrations of insulin lispro, specifically 5 IU (I-5), 10 IU (I-10), 15 IU (I-15), and 20 IU (I-20), as well as a control. The assessment of motility, plasma membrane integrity and DNA integrity was conducted using microscopy. Flow cytometry was used to assess acrosome integrity, mitochondrial membrane potential, and nitric oxide level. The preservation of motility, plasma membrane functional integrity, and acrosome integrity was significantly superior in the I-5 group compared to the control group. However, increasing levels of insulin had a detrimental impact on the integrity of the acrosome in both the I-15 and I-20 groups. The mitochondrial membrane potential was significantly elevated in the I-5 and I-10 groups compared to the control group. However, a statistically significant decrease was observed in both the I-15 and I-20 groups. We observed a significant decrease in nitric oxide levels in both the I-5 and I-10 groups as compared to the control group. The preservation of DNA integrity was significantly higher in the insulin containing groups compared to the control group. The study findings revealed that the inclusion of insulin lispro in drone semen extenders yielded beneficial outcomes for drone semen.

Cryopreservation and freezability of epididymal and ejaculated stallion spermatozoa

The aim of the present study was to compare the seminal attributes and freezability of the epididymal sperm recovered from castrated stallions and ejaculated semen collected through artificial vagina from the same stallions previously. The seminal quality parameters viz., viability, sperm morphology, plasma membrane functional integrity, acrosome integrity and mitochondrial membrane potential were studied, and recorded immediately after fresh semen collection/retrieval, before freezing, and after cryopreservation. Similar protocols were followed for freezing the epididymal and ejaculated semen. From the current study, it was observed that the sperm concentration of the epididymal recovered was significantly higher than that of fresh ejaculation. Other seminal parameters in thefresh sample, like total and progressive sperm motility, sperm membrane functional integrity, acrosome integrity and kinematic parameters like VSL, STR, and linearity were similar without any significant differences between both. However, mitochondrial membrane potential, BCF and linearity of the spermatozoa differed significantly between the groups after cryopreservation. From the current study, it can be concluded that the protocols used for cryopreserving the fresh ejaculates can be adopted for freezing the stallion epididymal spermatozoa without causing significant damage and through this method; elite stallion germplasm can be safely cryopreserved and conserved.

Cryopreservation and freezability of epididymal and ejaculated stallion spermatozoa

The aim of the present study was to compare the seminal attributes and freezability of the epididymal sperm recovered from castrated stallions and ejaculated semen collected through artificial vagina from the same stallions previously. The seminal quality parameters viz., viability, sperm morphology, plasma membrane functional integrity, acrosome integrity and mitochondrial membrane potential were studied, and recorded immediately after fresh semen collection/retrieval, before freezing, and after cryopreservation. Similar protocols were followed for freezing the epididymal and ejaculated semen. From the current study, it was observed that the sperm concentration of the epididymal recovered was significantly higher than that of fresh ejaculation. Other seminal parameters in thefresh sample, like total and progressive sperm motility, sperm membrane functional integrity, acrosome integrity and kinematic parameters like VSL, STR, and linearity were similar without any significant differences between both. However, mitochondrial membrane potential, BCF and linearity of the spermatozoa differed significantly between the groups after cryopreservation. From the current study, it can be concluded that the protocols used for cryopreserving the fresh ejaculates can be adopted for freezing the stallion epididymal spermatozoa without causing significant damage and through this method; elite stallion germplasm can be safely cryopreserved and conserved.

Cryopreservation of Pig Semen Using a Quercetin-Supplemented Freezing Extender.

Reactive oxygen species (ROS) produced during freeze–thaw procedures cause oxidative damage to the sperm, reducing fertility. We aimed to improve the post-thaw quality of pig sperm by quercetin (QRN) supplementation to reduce the cryodamage associated with the freeze–thaw procedure. Four equal aliquots of pooled boar semen were diluted with a freezing extender supplemented with different concentrations of QRN (0, 25, 50, and 100 µM) and then were subjected to cryopreservation in liquid nitrogen. Semen analysis was performed following 7 days of cryopreservation. Results demonstrated that the semen samples supplemented with 50 µM QRN significantly improved the post-thaw sperm quality than those subjected to other supplementations (p < 0.05). Semen samples supplemented with 50 µM QRN showed significantly improved plasma membrane functional integrity (47.5 ± 1.4 vs. 43.1 ± 4.1, 45.3 ± 1.7, and 44.1 ± 1.4) and acrosome integrity (73.6 ± 3.4 vs. 66.3 ± 2.4, 66.7 ± 3.6, and 68.3 ± 32.9) as compared to the control, 25 µM, and 100 µM QRN groups, respectively. The mitochondrial activity of the 50 µM QRN group was greater than control and 25 µM QRN groups (43.0 ± 1.0 vs. 39.1 ± 0.9 and 41.9 ± 1.0) but showed no difference with the 100 µM QRN group. Moreover, the 50 µM QRN group showed a higher sperm number displaced to 1 cm and 3 cm points in the artificial mucus than other groups. Therefore, supplementing the freezing extender with QRN can serve as an effective tool to reduce the magnitude of oxidative damage associated with sperm freezing.

Supplementation of melatonin to cooling and freezing extenders improves canine spermatozoa quality measures

BackgroundSperm freezing and cold storage are the two most common assisted reproductive technologies in the canine breeding industry. The freeze-thawing process causes significant detrimental changes in both sperm cell structure and function. Previous research has confirmed that excessive accumulation of un-scavenged free radicals (oxidative stress) plays an important role in the cryopreservation-induced damage to sperm cells. Also, the gradual accumulation of the free radicals during cold storage leads to a decline in the sperm quality markers. Melatonin is an endogenous neurohormone synthesized from tryptophan amino acid by pineal glands. Besides its several well-known physiologic roles, melatonin has a significant antioxidant potential through direct free radical scavenging properties. Therefore, the current study was designed to evaluate the potential in vitro protective properties of melatonin (0.5, 1, and 2 mM) on canine sperm cells after freezing or during long-term cold storage (9 days, 5 °C) on most important sperm in vitro fertility markers.ResultsMelatonin at 0.5, 1- or 2-mM concentrations could preserve significantly higher sperm total motility after 4 days of cold storage. However, only the 1- and 2 mM melatonin concentrations could result in better TM and PM values after 7 days of cold storage. Furthermore, melatonin supplementation could preserve higher sperm viability and acrosome integrity after 7 days of storage. Also, it could have significant protective effects on the cooled sperm DNA integrity. In the freezing section of the current research, melatonin at either 1- or 2-mM concentrations could not improve the sperm post-thaw TM and PM, whereas they improved sperm DNA integrity. Also, the post-thaw plasma membrane functional integrity and sperm velocity parameters were not affected by the treatment. Although DMSO (Dimethyl Sulfoxide) as the melatonin solvent could reduce the level of sperm lipid peroxidation and even improve the post-thaw sperm DNA integrity compared to the negative control, it reduced the post-thaw sperm progressive motility. However, the negative effects were reversed by concurrent melatonin supplementation at 1- and 2-mM concentrations.ConclusionThe addition of 1- or 2-mM melatonin to the canine sperm freezing and cooling media could improve sperm motility, viability, acrosome, and DNA integrity.

Positive effect of autologous platelet rich plasma on Saanen buck semen cryopreservation in non-breeding season

The objective of the current study was to evaluate the effect of autologous platelet-rich plasma (PRP) addition into soybean lecithin based extender on buck semen at post-thaw. Semen samples were collected from eight Saanen buck, and each semen sample was split into four equal aliquots and diluted with different concentrations of PRP supplemented extenders [no PRP (control), 0.5 × 107/ml PRP, 1 × 107/ml PRP, or 2 × 107/ml PRP]. Motility, plasma membrane functional integrity, acrosome integrity, mitochondrial membrane potential, DNA integrity and malondialdehyde concentrations (MDA) were measured and analyzed at post-thaw. The results showed that 2 × 107/ml PRP group had a positive effect on motility (62.41 ± 4.24), membrane functional integrity (71.11 ± 2.90), mitochondrial membrane potential (69.70 ± 1.99), DNA integrity (7.22 ± 0.93) and MDA levels (2.56 ± 0.73) at post-thaw (P < 0.05). The results of the study demonstrated that autologous PRP has a protective effect on cryopreservation of buck spermatozoa and the fertility effects are worthy of further study.

Effects of alpha-lipoic acid on ram semen cryopreservation and post-thaw life span.

The aim of the present study was to evaluate the effects of an alpha-lipoic acid-supplemented extender on ram semen at a post-thaw stage and after incubation (6hr). Semen samples were collected from five Kivircik Rams. Pooled semen was diluted with an egg yolk-based extender with different concentrations of alpha-lipoic acid (0.25mmol L-1 , 0.5mmol L-1 and 1mmol L-1 ) and without alpha-lipoic acid. Motility, plasma membrane functional integrity (HOST), acrosome integrity (FITC-Pisum sativum agglutinin) and DNA integrity (TUNEL) were assessed at post-thaw and 6hr after incubation of the frozen-thawed semen. At the post-thaw stage, 0.25mmol L-1 alpha-lipoic acid had a positive effect on sperm motility and plasma membrane functional integrity compared to the control group (p<.05). At the post-incubation stage (6hr), it was determined that the motility and plasma membrane functional integrity of the antioxidant groups were adversely affected compared to the control group (p<.05) and 1mmol L-1 dose of alpha-lipoic acid had a harmful effect on DNA integrity compared to the control group (p<.05). The results of the study demonstrated that alpha-lipoic acid has positive effects at post-thaw but have harmful effects on long term to ram semen.

Fluoride induced testicular toxicities in adult Wistar rats

Fluoride is essential for the development of teeth and bone but its excessive exposure causes reprotoxic effects. We have studied the graded effects of different doses of sodium fluoride (NaF) on 24 adult Wistar rats which were randomly divided into four groups (n = 6). All the rats were given normal diet prepared in our laboratory. Control (group I) rats were received vehicle only and treated rats (group II, III, and IV) were administered NaF orally at 10, 15, and 20 mg/kg/d doses, respectively, for 30 consecutive days. Gravimetric index of testis was decreased significantly in group III and IV rats but such changes were not observed in the accessory reproductive organs. Marked alterations in number, motility, viability, and plasma membrane functional integrity of caudal spermatozoa were noted in most of the treated groups. Noticeable alterations in testicular histoarchitecture were observed in group III and IV rats. Graded Changes of testicular redox homeostasis were noted by assessment of enzymatic (superoxide dismutase [SOD], catalase, glutathione s-transferase [GST], and glutathione peroxidase [GPx]) and nonenzymatic (malondialdehyde [MDA] and protein carbonyl [PC]) markers. Changes of testicular functional markers (acid phosphatase [ACP], alkaline phosphatase [ALP], and lactate dehydrogenase (LDH) activity) were noted in group III and IV rats. DNA fragmentation assay proved the possibility of DNA damage caused by oxidative stress, as evidenced by prominent trailing pattern of fragmented testicular genomic DNA from group III and IV rats. The study concludes that fluoride-mediated oxidative stress not only impedes the structural and functional facets of testis but also results in testicular DNA damage.

Effect of vitamin E supplementation on plasma membrane functional integrity in Surti buck semen preserved at refrigeration temperature

Effect of vitamin E supplementation on plasma membrane functional integrity in Surti buck semen preserved at refrigeration temperature

Soy lecithin as a potential alternative to powdered egg yolk for buck sperm cryopreservation does not protect them from mitochondrial damage

The aim of this study was to address whether soy lecithin (SL) was an effective non-penetrating cryoprotectant for buck sperm cryopreservation in the presence of seminal plasma. There was also an attempt to determine the optimal concentration of BHT as an antioxidant in powdered egg yolk (PEY) or in SL based media. Two ejaculates were collected from six bucks and mixed ejaculates were aliquoted into washed, using centrifugation procedures, and unwashed samples. In Experiment 1, washed sperm were re-suspended in PEY (15%) or SL (1%) media, while unwashed semen was only diluted in SL medium. In Experiment 2, washed and unwashed sperm were diluted in PEY and SL media, respectively, with there being different BHT concentrations (0.6, 2.0 and 5.0 mM). In both experiments, after 4 h of refrigeration, there were no differences neither in sperm viability nor plasma membrane functional integrity (HOST) between groups when there were evaluations using eosin-nigrosine staining. After thawing, however, there was a negative effect on motility of washed sperm preserved in SL media. Furthermore, results from cytometry evaluations indicated there was a larger population of thawed sperm with intact plasma (SYBR-14+/PI-) and acrosome (PE-PNA-) membranes, but inactive mitochondria (Mitotracker deep red-) when SL media were used. When there was BHT supplementation, there was only a slight enhancement of motility of spermatozoa preserved in PEY media with 5 mM BHT. In conclusion, when effectiveness and efficiencies are considered, PEY is the non-penetrating cryoprotectant that should be utilized for buck sperm cryopreservation.

Successful cryopreservation of honey bee drone spermatozoa with royal jelly supplemented extenders

The aim of the present study was to evaluate different concentrations of royal jelly (RJ) supplemented extenders for post-thawing quality of drone sperm. Semen samples were collected from sexually mature drones. Pooled semen was diluted with extender without RJ (control) or supplemented with different concentrations of RJ (1, 2, 4 or 8%). Sperm motility, plasma membrane functional integrity, and acrosomal integrity were evaluated. At post thaw, the highest sperm motility and acrosomal integrity rates were obtained in the RJ1 group. Functional integrity of sperm membrane was better preserved in the RJ1 and RJ2 groups compare to the other groups. The study shows that RJ supplemented extenders have beneficial effects on drone semen parameters. The results of the present study demonstrated advantage of using 1% RJ supplemented extender.

Effect of sperm pooling with seminal plasma collected in breeding or nonbreeding season on Saanen goat sperm cryosurvival.

The aim of this study was to determine the effects of both the removal of seminal plasma (SP) and the pre-freezing addition of seminal plasma collected during the breeding or nonbreeding season on goat sperm survival after thawing. Semen samples were pooled. One aliquot of pooled semen was used as a control group. Four aliquots were then centrifuged, and the SP was removed in Group I, pipetted but not removed in Group II, removed and then pooled for animals collected in the breeding season in Group III and removed and pooled for animals collected in the nonbreeding season in Group IV. Group samples were frozen and then were assessed for rates of sperm motility, plasma membrane functional integrity hypo-osmotic swelling test (HOST), defective acrosomes (FITC-PSA), DNA fragmentation (TUNEL) and mitochondrial membrane damage (Rhodamine 123). The results showed that pre-freezing addition of SP collected in breeding season maintained post-thaw sperm characteristics at 0hr better than SP removal group, but removing seminal plasma showed positive effects on spermatozoa, as incubation time increased to 5hr. In conclusion, the pre-freezing addition of seminal plasma did not maintain post-thaw goat sperm characteristics as successfully as in the groups with seminal plasma removed after an incubation period.

Positive effect of BSA supplemented soybean lecithin based extender on liquid storage of ram semen at 5C

The aim of the study was to evaluate different BSA doses in soybean-lecithin based extenders for liquid storage of ram semen. Pooled semen was divided into four equal volumes and diluted in a single-step dilution method with BSA supplemented (BSA5, BSA7.5, BSA10 groups supplemented 5mg/mL, 7.5mg/mL, 10mg/mL BSA respectively) and BSA-free (control) extenders. Semen samples were assessed for sperm motility, plasma membrane functional integrity using hypo-osmotic swelling test (HOST), damaged acrosome using FITC-Pisum sativum agglutinin (PSA-FITC) at 0, 24, 48 and 72h of storage at 5 °C. Antioxidant activities of groups at 0 and 72h were measured with glutathionine (GSH) concentration, membrane lipid peroxidation status was analyzed using the malondialdehyde (MDA) concentration. Antioxidant capacity of extender also was measured with 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and Dichloro-dihydro-fluorescein diacetate (DCFH-DA) assays. In the study, BSA (5mg/mL) supplementation has beneficial effect on ram sperm parameters at all evaluation times. In addition, the increasing doses of BSA supplementation has a negative effect on sperm parameters especially 48 and 72h of storage

Impact of arsenic(V) on testicular oxidative stress and sperm functional attributes in Swiss albino mice.

The arsenic (As) is a multi system effector including reproduction. The present study examined the association of graded doses of As(V) on testicular microenvironment and sperm function in mice. Thirty-six adult male mice were randomly assigned to six groups (n = 6). Group A served as control without test chemical. The groups B, C, D, E, and F were administered graded doses of 10, 25, 50, 100, and 200ppm As(V), respectively, through drinking water for 40days. A dose-dependant significant (P < 0.05) decrements were observed in epididymal sperm kinematic attributes (progressive motility, rapid, fast progressive, VCL, VSL, VAP, LIN, STR, WOB and TYPE A (STR >80%, ALH 2.5μm) by CASA), viability, plasma membrane functional integrity, and mitochondrial membrane potential which were associated with insignificant decrease in serum testosterone levels. The histoarchitectural studies of testes showed progressive loss of spermatozoa concentration in the seminiferous tubules as the As(V) dose increased. The mice exposed to As(V) had an increase in the As accumulation, protein carbonylation, and lipid peroxidation levels associated with alterations in SOD, CAT, and GST activities in the testes. In conclusion, higher doses of As(V) (more than 50ppm) were found to be testicular toxicants which impaired semen quality by inducing oxidative stress in the testicular microenvironment.

Cryopreservation of ram semen with antioxidant supplemented soybean lecithin-based extenders and impacts on incubation resilience

The scope of this study was investigation the affects of various antioxidants on 1% soybean lecithin-based semen extenders for ram semen cryopreservation. Ejaculates, collected via electrically stimulated ejaculation, that have a thick consistency, rapid wave motion (3–5 on a 0–5 scale) and >75% initial motility were pooled. The pooled samples were split into four equal aliquots as 5 mM Methionine, 5 mM Cysteamine, 1 mM Cysteine and a sample of antioxidant-free control group. Each sample group was diluted to a ratio of 1/5 (semen/extender, v/v) as final concentration and two step dilution method was used for cryopreservation. Extender groups were assessed for sperm motility, plasma membrane functional integrity using hypoosmotic swelling test (HOST), damaged acrosome using FITC-Pisum sativum agglutinin (PSA-FITC) and DNA integrity using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). Semen samples also incubated for 6 h in humidified air with 5% CO2 at 39 °C to evaluate post-thaw incubation resilience of semen characteristics. The results showed that freezing and thawing procedures had negative effects on motility (P < 0.05), plasma membrane integrity (P < 0.05) and acrosomal integrity (P < 0.05). After 6 h of incubation time, the Cysteine supplemented extender group yielded significantly higher results than other extender groups in terms of spermatological parameters. Furthermore MDA levels in the antioxidant groups were lower than control group (P < 0.05). Nevertheless, there were no significant differences among antioxidant groups.

L-CARNITINE ENHANCING ROLES ON BUFFALO SEMEN FREEZABILITY, ULTRA STRUCTURE AND FERTILIZING POTENTIALS

Recently, male impact has been recognized as a momentous cause of infertility. Male infertility isn’t an entity but it evokes a variety of mechanisms and explanations to understand it. In between, a biochemical semen additive that improves or overcome certain semen situations so can enhance or treat semen fertility and infertility, respectively. L-carnitine is one of these biochemical semen additives. Till now, the exact effects of L-carnitine on buffalo semen processing outcomes haven’t been discovered. The current study aimed to clarify L-carnitine roles during buffalo semen cryopreservation. Semen samples were obtained from six fertile buffalo bulls (aged 3 to 5 years). Weekly, two consecutive ejaculates were collected from each bull for successive six weeks using artificial vagina. The ejaculates were pooled to eliminate samples variability. Semen samples were extended with Tris-based extender supplemented with different concentrations of L-carnitine (0.01, 0.05 and 0.1 mg/ml) Vs. Tris-based extender only (control). Then they were processed to cryopreservation and thawing to assess different semen characteristics. It had been found that L-carnitine (0.05mg/ml) significantly (p < 0.05) improved post thawing motility, viability index, acrosomal integrity, in vitro fertilization, blastocyst and conception rates of treated buffalo semen (57.00±2.55%, 126.5±8.33, 14.40 ± 1.43%, 56.45%, 15.79% and 64.71%, respectively), compared with control (42.00±2.56%, 87.00±10.43, 23.80±1.86%, 36.53%, 2.13% and 42.86%, respectively). Conclusion: L-carnitine supplementation to buffalo semen extender significantly enhanced its characteristics and protecting its plasma membrane and mitochondrial functional integrity. Moreover, the preceding results focusing more light on the potential roles of L-carnitine in regulating male fertility and infertility.

Freeze-dried egg yolk based extenders containing various antioxidants improve post-thawing quality and incubation resilience of goat spermatozoa

The aim of this study was to evaluate different antioxidants-supplemented freeze-dried egg yolk based extenders for the post-thawing quality and incubation resilience of goat spermatozoa. Pooled semen were diluted in a two-step dilution method to a final concentration of 1/5 (semen/extender) in control and antoxidant supplemented freeze-dried extenders (methionine, cysteamine and butylated hydroxytoluene). Semen samples were assessed for sperm motility, plasma membrane functional integrity using hypoosmotic swelling test (HOST), damaged acrosome using FITC-Pisum sativum agglutinin (PSA-FITC) and DNA integrity using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). Membrane lipid peroxidation status was also analyzed using the malondialdehyde (MDA) concentration. In the study, antioxidant supplemented freeze-dried egg yolk based extenders have beneficial effect on goat sperm parameters. In addition, we achieved a higher quality in post thawed goat semen even after 6 h incubation when the extender was supplemented by 5 mM BHT or cysteamine.