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Effects of astaxanthin supplementation on the sperm quality and antioxidant capacity of ram semen during liquid storage

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... In recent years, many studies have been focused in the use of exogenous antioxidants to ram semen extenders to diminish the negative effect induced by ROS (Çoyan et al., 2010;Mata-Campuzano et al., 2014;Moradi, Malekinejad, Farrokhi-Ardabili & Bernousi, 2013;Allai et al., 2015;Fang et al., 2015, Rather, Islam, Malik & Lone, 2016Allai, Benmoula, da Silva, Nasser & El Amiri, 2018). The main antioxidants are extracts from the leaves, seeds and roots of plants due to their high content of polyphenols, flavonoids, carotenes, gallic acid, tannins and essential oils (Zhong & Zhou, 2013). ...
... When other antioxidants have been tested in liquid ram sperm, contradictory results have been obtained, suggesting that results could be influenced by other factors such as the male, extender, antioxidant or concentration (Silva, Cajueiro, Silva, Soares & Guerra, 2012). Assorted results are found in literature showing positive (using catalase, methionine, astaxanthin or vitamine C) (Câmara et al., 2010;Bucak, Coyan, Oztürk, Güngor, & Omür, 2012;Fang et al., 2015) or not positive effects (such as glutathione, ergothioneine, reduced glutathione or trolox) (Bucak & Tekin, 2007;Mata-Campuzano et al., 2014) of antioxidants on the sperm motility. In this sense, some authors suggested that the addition of natural substances having antioxidant effect acts as double-edged swords, since high exogenous antioxidants may disrupt redox balance and acts like a pro-oxidant by activating pathways such as increasing pro-inflammatory mediators production, nitrosylation of proteins, proglycation effect and endocrine disrupting activities (Bouayed & Bohn, 2010). ...
... By contrast, control In a recent study, it has been reported that the addition of HT, DHPG and the combination of both in criopreservated ram sperm did not offer neither better nor detrimental effects on the kinetic and velocity parameters in comparison with the control group (Arando et al., 2019), probably because lower antioxidant concentrations were added to the sperm samples than in the present study. In the same way, when antioxidants such as catalase, glutathione or astaxanthin, were added to the sperm dilution media during liquid storage, no significant differences were observed for kinematic parameters (Câmara et al., 2010;Fang et al., 2015) suggesting that factors aside from oxidative effects, such as reduced energy production or metabolism, may contribute to their decline during storage at 4ºC. ...
Article
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The aim of the present study was to evaluate the effect of the addition of two olive oil‐derived antioxidants, hydroxytyrosol (3,4‐dihydroxyphenylethanol, HT) and 3,4‐dihydroxyphenylglycol (DHPG), on ovine semen during liquid storage at 5ºC and 15ºC. Semen was collected, pooled, diluted and then divided into aliquots supplemented with different concentrations (5 μg/ml, 10 μg/ml, 50μg/ml and 100 μg/ml) of HT, DHPG and a mixture (MIX) of both antioxidants. Sperm motility characteristics were assessed in the different samples at 0, 6, 24, 48, 72 and 96 h after cooling and a fertility trial was also conducted. The results showed that the antioxidant addition did not significantly improve total and progressive motility in ovine cooled sperm maintained at 15º or 5ºC. However, in samples stored at 5ºC, LIN (48, 72, 96 h), STR (0h) and WOB (0, 48, 72, 96h) values significantly decreased in comparison with control treatment when high antioxidant concentrations were added (MIX100 or HT100). When samples were maintained at 15ºC, MIX50 showed significantly higher VCL values than the control treatment after 6h cooling, and MIX100 showed significantly lower VCL values at 96h after cooling. According to the artificial insemination trial, no significant differences were observed when antioxidants were added. In conclusion, the use of HT and DHPG showed small impact in sperm motility and fertility was not affected (nor detrimentally nor positively) when insemination was carried out using antioxidant‐supplemented liquid sperm.
... The ASX compound is a xanthophyll carotenoid that has antioxidant, anti-inflammatory and anti-cancer properties (Ambati et al., 2014). The inclusion of ASX in semen extenders led to improvements in values for the sperm variables of liquid stored ram (Fang et al., 2015) and FT bull (Farzan et al., 2014) semen, and there were beneficial effects of oral administration (Comhaire et al., 2005) or in vitro supplementation (Andrisani et al., 2015) on human sperm quality and fertilizing capacity. In addition, there was a beneficial effect on liquid stored boar semen (Basioura et al., 2018), however, there have been no reports about effects of ASX on cryopreserved boar semen. ...
... Based on results from previous studies, ASX can scavenge hydroxyl radicals (Hama et al., 2012) and penetrate biological membranes, protecting the fatty acids from lipid peroxidation (Donà et al., 2013). With ASX supplementation of extenders at two different concentrations (2 μM and 4 μM), there is a reduced malondialdehyde production and lesser ROS concentrations in liquid stored (4°C) ram semen at the end of the 72 h of storage (Fang et al., 2015), while there was a similar beneficial effect on lipid peroxidation of the sperm membrane when ASX (0.5 μM) was added to bull sperm, which was diluted in an EY free cryopreservation extender (Farzan et al., 2014). Furthermore, ASX supplementation to in vitro maturation and culture medium resulted in an inhibition of lipid peroxidation and subsequently improvements in the blastocyst and embryo development in pigs and cattle when there were heat stress conditions prevailing at the time of oocyte collections (Kuroki et al., 2013;Do et al., 2015). ...
... It, therefore, could be assumed when there were these conditions that a possible interaction between ASX and EY was minimized. Based on results from previous studies, ASX has an antioxidant effect on liquid stored ram (Fang et al., 2015) or bull cryopreserved sperm (Farzan et al., 2014) regardless of whether extenders contain EY. These inconsistent results could indicate the need for further investigation to elucidate any possible interaction between antioxidants and EY-containing extenders that could be related to spermatozoa-and/or SP-specific species differences. ...
Article
The aim of the study was to determine whether the presence of astaxanthin (ASX) protects boar spermatozoa against damage related to cryopreservation. Pooled ejaculates extended in Beltsville Thawing Solution (BTS) were used. Three experiments were conducted: 1) sperm samples were pre-incubated overnight (17 °C) with ASX (0, 0.5, 5, 15 μM) prior to freezing and then frozen using cooling and thawing extenders supplemented with ASX (0, 0.5, 5, 15 μM); 2) sperm samples were treated with ASX (0, 0.5, 5, 15 μM) only during overnight pre-incubation (17 °C) prior to cryopreservation; and 3) a thawing extender was supplemented with ASX (0, 0.5, 5, 15 μM). The groups were as follows: control (C; no treatment), ASX 1 (0.5 μM), ASX 2 (5 μM) and ASX 3 (15 μM). Total (TM) and progressive (PM) motility was analyzed using CASA, while sperm viability, reactive oxygen species generation, lipid peroxidation and apoptoticlike changes were analyzed using flow cytometry. Sperm variables were evaluated prior to freezing as well as 30 and 150 min after thawing. In Experiment 1, the values of TM and sperm viability post-thaw were less in the ASX 3 than C group. In Experiment 2, there was no effect of ASX on any of the sperm variables evaluated, while in Experiment 3, apoptotic-like changes were less in the ASX 1 than C group. In conclusion, there was a subtle beneficial effect on cryopreserved boar spermatozoa after addition of ASX to thawing media.
... It can be concluded from the study that supplementation of AX to the semen extender assisted in maintaining or protecting the spermatozoa from damage during storage at 5 °C. membrane integrity (p≤0.05) in rams diluted semen during the storage period of 72 hours at 5 °C (Fang et al., 2015). Malondialdehyde (product of lipid peroxidation) and reactive oxygen species (ROS) levels also decreased (p≤0.05) ...
... Malondialdehyde (product of lipid peroxidation) and reactive oxygen species (ROS) levels also decreased (p≤0.05) markedly during 72 hrs of storage at 5 °C (Fang et al., 2015). Sperm membrane is prone to free radical attack due to the high content of phospholipids (Almbro et al., 2011). ...
... The lower level of CAT and SOD concentration in extended semen also indicates the antioxidant activity of AX. Similarly, Fang et al., (2015) and Farzan et al., (2014) also reported the improvement of sperm quality and reduction of malondialdehyde and ROS in AX supplementation during storage of ram and bull extended semen at 5 °C for 72 hours, respectively. AX is a lipid soluble pigment with potent inside and outside antioxidant properties gives overall protection (McNulty et al., 2007). ...
... Many studies reported that ASX is a promising antioxidant providing important biological properties, such as anti-inflammatory and anticancer actions (Ambati, Phang, Ravi, & Aswathanarayana, 2014). Also, ASX improved sperm parameters and fertility in human (Comhaire, El Garem, Mahmoud, Eertmans, & Schoonjans, 2005), ram (Fang et al., 2015) and bull (Farzan, Chamani, & Varnaseri, 2014). Nevertheless, the effect of ASX on boar semen has not been investigated, yet. ...
... In accordance with our results, ASX supplementation to bull and ram semen significantly increased the values of total and progressive motility (Fang et al., 2015;Farzan et al., 2014). Comhaire et al. (2005) demonstrated that ASX could positively affect male fertility by improving sperm motility. ...
... A showing an increase in VSL between 0 and 24 hr. However, in ram semen, ASX had no effect on VCL, VSL and VAP (Fang et al., 2015). Broekhuijse et al. (2012) indicated that VSL is correlated with the total number of born piglets. ...
Article
Contents The aim of the study was to investigate the effect of the antioxidant astaxanthin on boar semen. Twenty ejaculates from 10 boars (two ejaculates/boar) were extended and split in three groups: semen control (SC), solvent control (C; semen with dimethyl sulfoxide, the diluent of astaxanthin) and semen with astaxanthin (A) in concentration 0.5 μmol/L. Sperm quality parameters (motility and kinetics, morphology, viability, functional integrity of sperm plasma membrane by Hypo‐Osmotic Swelling Test [HOST] and DNA integrity) were assessed at 0, 24 and 48 hr of storage at 17°C (experiment I), before (0 hr) and after (1 hr) of sperm thermal resistance assay at 37°C (experiment II) and finally before (0 hr) and after (1 hr) sperm in vitro incubation (38.5°C, 5% CO2, maximum humidity [experiment III]). In experiment I, group A performed overall better than group SC and as a tendency better than group C regarding viability. Total motility, rapid spermatozoa and HOST remained constant across time in group A, whereas they decreased in the remaining groups. In experiment II, regarding motility and viability, group A displayed better results across time than the other two groups. In experiment III, viability and total motility decreased in groups SC and C, while in group A, these parameters were not significantly different between the examination time points. In conclusion, astaxanthin has a beneficial and protective effect on boar semen quality under the investigated conditions.
... One of efforts conducted to solve this problem is antioxidant treatments of sperms during preservation. Previous studies showed that exogenous antioxidant treatments improved the vitality and motility of sperms in bulls (Sariözkan et al., 2009) and rams (Asadpour et al., 2012;Fang et al., 2015). A study by et al. (1993) reported that β-carotene could reduce destructive ability of oxygen singlet by physically accepting energy from the electron-rich structure of reactive oxygen species. ...
... Finally, frozen semen was thawed in a 37 °C waterbath (Memmerth, Germany) for 30 seconds and assessed for motility, viability, acrosomal integrity, and plasma membrane integrity (Fang et al., 2015). ...
Article
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Sperms are vulnerable to oxidative stress due to the high proportion of polyunsaturated fatty acids within their membranes. This condition could decrease sperms quality during preservation. ß-carotene is among antioxidants able to neutralize reactive oxygen species, natural by-products of oxygen metabolism in sperms. This study was done to investigate the capability and protective effect of this antioxidant on motility, viability, acrosomal integrity, and plasma membrane integrity of frozen-thawed sperms of Aceh swamp buffalo. The study was conducted using completely randomized design with four treatments and five replications. Fresh semen was diluted in egg-yolk tris-based extenders with the addition of antioxidant of 0% (w/v) (control, P0), 0.625% (w/v) (P1), 1.25% (w/v) (P2), and 2.5% (w/v) (P3) prior to freezing. Variables of sperms quality examined were motility, viability, acrosomal integrity, and plasma membrane integrity of semen after dilution, equilibration, and thawing. Semen having the best post-thawing quality after ß-carotene treatment was used for artificial insemination (AI) for accessing its pregnancy rate. The results showed that the addition of 0.625% (w/v) ß-carotene (P1) resulted in the highest motility, viability, acrosomal integrity, and plasma membrane integrity. Pregnancy rate of buffalo inseminated with semen treated with 0.625% ß-carotene (P1) was 50%. In conclusion, administration of 0.625% ß-carotene was able to maintain and could protect motility, viability, acrosomal integrity, and plasma membrane integrity of frozen sperms of Aceh swamp buffalo that are good for artificial insemination.
... There are limited studies regarding the effect of CX on semen quality in ram. Although astaxanthin and lycopene carotenoids have been used as antioxidant in semen extender and significantly improved the progressive sperm motility and viability during the storage period in ram (Fang et al., 2015;Uysal & Bucak, 2007) and boar (Lee & Kim, 2018), the morphological abnormal sperm percentage significantly decreased in MLT and CX group, indicated their protective effect on spermatozoa during storage of liquid semen. These findings are also favoured with the previous studies that support the role of MLT and CX in protection of spermatozoa during storage period (Khalifa, 2017;Najafi et al., 2019). ...
... Khalifa (2017) and Dai et al. (2019) also observed significant positive effect of MLT on plasma membranes integrity in rams. Astaxanthin with lycopene also showed significant effect on membrane integrity of spermatozoa (Fang et al., 2015;Tuncer et al., 2014). ...
... However, the current study investigates the protective effects of Ast, as a lipid-soluble carotenoid against Cdinduced toxicity in rooster testis and reduced sperm quality. Recently, the red carotenoid pigment Ast, a potent antioxidant without provitamin-A activity present in some marine organisms (Lai et al., 2004), has been used in numerous studies to improve sperm quality (Mansour et al., 2006;Fang et al., 2015;Tizkar et al., 2015b;Vahidinia et al., 2017). Furthermore, it has been reported in the literature that the antioxidant activity of Ast is approximately 10 times higher than other carotenoids such as zeaxanthin, lutein, canthaxanthin, and b-carotene and about 100 times than that of a-tocopherol (Miki, 1991). ...
... With the rapid development of modern industries, reproductive impairment caused by environmental pollution should never be neglected. Many experiments have used Ast to enhance sperm quality in human (Dona et al., 2013;Andrisani et al., 2015), ram (Fang et al., 2015), fish (Mansour et al., 2006;Tizkar et al., 2015a, b), and rat (Vahidinia et al., 2017) and reported that this antioxidant can improve sperm capacitation. In this study, we investigated the protective effects of Ast against LPO, and changes in enzymatic (MDA, CAT, SOD, TAC, and GPX) defense systems induced by Cd in rooster sperm. ...
Article
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The protective role of astaxanthin nanoparticles (Ast NPs, 25 mg/kg p.o) against cadmium (Cd, 1 mg/100 g b.w. SC), a known inductor of lipid peroxidation and changes in the antioxidant defense system in the Ross 308 breeder roosters sperm, was examined. Sperm motility (computer-assisted sperm motility analysis), membrane integrity (hypoosmotic swelling test), viability, total abnormality, and enzymatic parameters were assessed after thawing. The testis/body weight (mg/kg) ratio and HE staining results of testis were also performed. The obtained results showed that Cd induced detrimental effects on testis and sperm, while Cd treated by Ast NPs (Cd Ast) diminished this change compared to the Cd group. Cd-treated group resulted in significantly (P < 0.05) lowest total (37.29 ± 2.46) and progressive (5.84 ± 0.47) motility and decreased antioxidant enzyme activity (CAT, TAC, and GPx), as well as producing a significant (P < 0.05) decrease in testis weight (mg) compared to the control group. Treatment with Ast NPs (Ast NPs + Cd) had reversed Cd-induced changes in the antioxidant defense system and significantly prevented Cd-induced testis damage. In conclusion, the results of our work suggest that Ast NPs at 25 mg/kg act as a potent antioxidant in protecting rooster testes against oxidative stress induced by Cd.
... There are limited studies regarding the effect of CX on semen quality in ram. Although astaxanthin and lycopene carotenoids have been used as antioxidant in semen extender and significantly improved the progressive sperm motility and viability during the storage period in ram (Fang et al., 2015;Uysal & Bucak, 2007) and boar (Lee & Kim, 2018), the morphological abnormal sperm percentage significantly decreased in MLT and CX group, indicated their protective effect on spermatozoa during storage of liquid semen. These findings are also favoured with the previous studies that support the role of MLT and CX in protection of spermatozoa during storage period (Khalifa, 2017;Najafi et al., 2019). ...
... Khalifa (2017) and Dai et al. (2019) also observed significant positive effect of MLT on plasma membranes integrity in rams. Astaxanthin with lycopene also showed significant effect on membrane integrity of spermatozoa (Fang et al., 2015;Tuncer et al., 2014). ...
... There are limited studies regarding the effect of CX on semen quality in ram. Although astaxanthin and lycopene carotenoids have been used as antioxidant in semen extender and significantly improved the progressive sperm motility and viability during the storage period in ram (Fang et al., 2015;Uysal & Bucak, 2007) and boar (Lee & Kim, 2018), the morphological abnormal sperm percentage significantly decreased in MLT and CX group, indicated their protective effect on spermatozoa during storage of liquid semen. These findings are also favoured with the previous studies that support the role of MLT and CX in protection of spermatozoa during storage period (Khalifa, 2017;Najafi et al., 2019). ...
... Khalifa (2017) and Dai et al. (2019) also observed significant positive effect of MLT on plasma membranes integrity in rams. Astaxanthin with lycopene also showed significant effect on membrane integrity of spermatozoa (Fang et al., 2015;Tuncer et al., 2014). ...
Article
Antioxidants are used to minimize oxidative stress during liquid semen storage. The main aim of current study was to elucidate effect of supplementing melatonin and canthaxanthin in Tris-based extender could enhance seminal quality of ram at 4°C up to 72 h. A total of 48 ejaculates were collected from breeding Magra rams (n = 8) and were preliminarily subjected for various macroscopic and microscopic semen evaluation tests. These ejaculates were pooled and divided into three equal aliquots. Two aliquots were diluted (1:10) using extender encompassing final concentration of 1mM melatonin and 25 µM canthaxanthin and stored at 4°C. Third aliquot with extender only was kept as control. Structural and functional seminal changes were observed at different time points of preservation. Results revealed that mean values for progressive sperm motility, viability and total antioxidant capacity were significantly higher (p < 0.05) in melatonin group while hypo-osmotic swelling test was significantly (p < 0.05) higher in canthaxanthin group. Total sperm abnormalities and malondialdehyde levels were significantly (p < 0.05) lower in both treatment groups indicating their antioxidant efficacy in protection of spermatozoa from oxidative stress. Results of study indicated that supplementation of these antioxidants to ram semen could be used to enhance storage life of liquid semen at 4°C up to 72 h.
... Foote et al. (2002) have described that the taurine can cross the sperm plasma membrane and protect it. Previous studies reported that the values of ram and bull individual motility were improved by astaxanthin adding (Fang et al., 2015). Basioura et al., (2017) found the addition of 0.5 μМ / L of astaxanthin to boar semen extender improved individual motility during 1 hour of sperm incubation at 5°C. ...
... compared with the control treatment. Whereas, the progressive, path, and curvilinear velocities were not affected (Fang et al. 2015). By entering the membrane and creating a capacitation-like membrane alteration, AST allows Tyr-P of the head. ...
Article
The future fertility of cancer patients after treatment is a serious concern amongst young cancer patients. Such chemotherapeutic agents as Methotrexate (MTX), a folic acid antagonist can cause long-term or permanent gonadal toxicity in male patients. Oxidative stress is a contributing mechanism to MTX-induced testicular damage. In the past decade, several studies have investigated the use of natural antioxidant agents to prevent the side effects of MTX. Astaxanthin (AST), a red-orange xanthophyll carotenoid, owns various clinical benefits and pharmacological effects including antioxidant, anti-tumor, anti-cancer, anti-diabetic, and anti-inflammatory properties. Since there has been no study so far in the literature on the effects of AST on MTX-induced testicular dysfunction, the present study evaluated the ameliorating effect of AST against MTX-induced testicular dysfunction. Biometric and semen parameters as well as, biochemical markers of oxidative stress were measured. Compared to the control group, MTX-treated group showed a significant decrease in sperm count, motility, viability, morphology, superoxide dismutase (SOD) and catalase (CAT) activities, and a significant increase in MDA levels. Pretreatment with AST for six consecutive days before MTX administration prevented these oxidative parameters. Thus, the current results suggest that MTX seems to impair the male fertility through oxidative damage in one hand, and AST pretreatment might improve the male fertility by preventing oxidative stress-induced fertility disorders, on the other hand. However, AST pretreatment might also protect germ cells against MTX-induced oxidative stress.
... DCFH-DA, an oxidation-sensitive fluorescent probe, was used to analyze ROS generation. 21 Sperm were treated as described in section 2.3 and then incubated with 5 µmol l −1 DCFH-DA for 30 min at 37°C. The cellular mean fluorescence intensity (MFI) was measured using flow cytometry. ...
Article
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Polychlorinated biphenyls (PCBs) are common environmental contaminants that represent a considerable risk to reproductive toxicity in exposed human populations. Although some experimental studies have suggested an association between the levels of PCBs and semen quality, the direct effects of PCBs on human sperm parameters remain largely unexplored. To this aim, a short-term in vitro incubation experiment that better imitated the putative exposure of sperm to Aroclor 1254 (a commercial PCB mixture) in male reproduction tissue was conducted. Human sperm were incubated with various concentrations (0, 1, 5, or 25 mg l-1 ) of Aroclor 1254 for different amounts of time (3 and 6 h) in vitro. Sperm motility parameters were analyzed with computer-assisted sperm analysis (CASA). The proportion of sperm with high mitochondrial membrane potential (ΔΨm) and the levels of intracellular reactive oxygen species (ROS) were detected to explore the probable cause of sperm impairment. Human sperm exposed to continuous Aroclor 1254 exhibited: (i) reduced sperm motility and kinematic parameters, (ii) a proportion of sperm with high ΔΨm that decreased in a dose-dependent manner (P < 0.05), and (iii) increased levels of ROS compared with controls (P < 0.05). In conclusion, Aroclor 1254 can decrease sperm motility, which may culminate in increased ROS and general mitochondrial dysfunction, thus affecting the fertilization potential of sperm. Our findings suggest a broader understanding of the effect of Aroclor 1254 on human sperm.
... It is probable that the disruption caused by the concentration of 25 mM in the kinetic variables (TM, WOB, and ALH) could not cause impairment to the integrity of membranes. Alternatively, we can cite a study where astaxanthin, a carotenoid that like canthaxanthin belongs to the class of xanthophylls, was used to supplement an extender egg yolk based on the concentrations of 2 and 4 mM, which improved the integrity of the plasma membrane and the total and PM of ram sperm after 72 hours of liquid storage at 4°C. 44 It is noteworthy that these results were observed in cooled semen that unlike our experiment, did not suffer intense osmotic shock, as noted in the freezing and thawing processes. In addition, the gametes in this study were submitted to a long period of storage (72 hours), which manifested through elevation in the ROS and malondialdehyde (MDA) levels, which can explain the protective effect of astaxanthin to spermatozoa. ...
Article
The addition of antioxidants to semen cryopreservation extenders has been employed for combating oxidative damage. This work aimed to evaluate the addition of carotenoid canthaxanthin to a cryopreservation extender of ram semen. Three breeder rams were used and, after semen collection, with 48-hour intervals between collection, the samples were included in the pool formation (n = 6). The experimental groups comprised 0 (control), 0.1, 1, 10, and 25 μM of canthaxanthin. After thawing (37°C/30 s) and incubation at 37°C for 2 hours, semen aliquots from each group were evaluated for sperm kinetics (CASA), the integrity of the plasma and acrosomal membranes (iPAM), intracellular reactive oxygen species (ROS) production, and lipid peroxidation (LPO) by flow cytometry associated with the image. The control group and canthaxanthin 1 μM after incubation at 37°C for 2 hours showed increases of curvilinear velocity and amplitude of lateral head displacement with decreases of linearity, straightness, and wobble (p < 0.05), which were not observed for the canthaxanthin 10 and 25 μM. The supplementation of a Tris-egg yolk extender with canthaxanthin had no effect on the iPAM, intracellular ROS production in viable spermatozoa, or LPO. In conclusion, supplementation with 10 and 25 μM of canthaxanthin in a Tris-egg yolk extender used for ram semen cryopreservation is able to protect ovine sperm from kinetic changes after incubation at 37°C for 2 hours post-thawing.
... For cooled stored ram semen, egg yolk has been used in concentrations as great as 40% (v/v) in Tris-buffered extenders (Câmara et al., 2011;Fang et al., 2015;Rateb, 2018;Amini et al., 2019;Abadjieva et al., 2020) or in a proportion of 5-21% (v/v) supplemented to skim milk-based extenders (Paulenz et al., 2002;Fernandez-Abella et al., 2003;Kulaksiz et al., 2012;Galarza et al., 2020). Ram sperm maintained approximately 30% sperm motility after 16 days of storage at 5 • C in Tris-based extenders supplemented with trehalose and 10% (v/v) egg yolk (Lopez Saez et al., 2000). ...
Article
This review is part of the Festschrift in honor of Dr. Duane Garner and provides an overview of current techniques in cooled storage of semen from livestock animals such as camelids, goats, and sheep. Facing worldwide environmental changes and a trend towards more conscious and healthy eating behaviors, the development of a stable animal breeding industry is a significant challenge for the near future. In the present review, factors influencing semen handling in camelids, goats and sheep are described and relevant methods as well as current trends to improve liquid-storage of cooled semen are discussed, including extenders, additives, cooling rates, and storage temperatures. The species-specific physiology and resulting challenges are taken into consideration. While the main problem for camelid semen processing is the relatively greater viscosity as compared with that of some other animals, the deciding factor for successful artificial insemination (AI) in goats and sheep is the site (i.e., cervical or vaginal) of semen placement in the reproductive tract. Due to the type of cervical anatomy, the penetration of the cervix when using AI instruments is rather difficult. Furthermore, the seminal plasma of small ruminants affects the interaction with milk-based extenders and egg yolk which results in species-specific regimens for cooled liquid-preservation. Comparing all three species, the greatest pregnancy rates were obtained by AI with goat semen after cooled liquid-storage for several days.
... Astaxanthin fed rats have shown a substantial reduction in ROS production in semen and improved male fertility (Mortazavi et al. 2014). Semen extender added with astaxanthin helped in improving the sperm quality and function of ram by reducing of malondialdehyde and ROS during storage (Fang et al. 2015) and bull (Farzan et al. 2014, Soren et al. 2017 semen at 5 °C for 72 hours. During production of miniature pig semen straw, adding of astaxanthin to frozen storage solutions, prevented damages during freeze-thawing process and maintained quality of sperms and improved fertility rate (Lee and Kim, 2018). ...
... In an in vitro study, the motility of sperm in AST (2 and 4 micromolar) treated rams' cells increased significantly, while curvilinear velocity, progressive velocity, and path velocity were not affected. Supplementing semen extender with AST can enhance semen preservation via protection of plasma membrane integrity [240]. AST pretreatment also safeguarded germ cells against methotrexate-induced OS. ...
Article
Astaxanthin (AST) is a potent lipid-soluble keto-carotenoid with auspicious effects on human health. It protects organisms against a wide range of diseases with excellent safety and tolerability. Various imperative biological activities in vitro and in vivo models have been suggested for AST. This review article is focused on the therapeutic potentials, biological activities and benefical health effects of AST. The pharmacological mechanisms of action of AST in the treatment and prevention of the peripheral and central nervous system diseases was also reviewed to provide new insights to researchers. Finally, we suggested a novel hypothesis for the mechanism of action of AST in neuropathic pain following spinal cord injury.
... Our results revealed that addition of resveratrol to the extender has a potential to increase SOD and GSH levels in ram spermatozoa and inhibits MDA production during cooling storage at 5 C. Similar results were reported after addition of other antioxidants as ascorbic acid, butylated hydroxyl-toluene and taurine to ram semen extender; these antioxidants were shown to protect spermatozoa from oxidative harm during three days of cooling storage [22]. Our results are consistent with previous studies that confirmed that antioxidants supplementation ameliorates ram spermatozoa quality during cooling storage [20,23,43]. Similar results were also reported for frozen-thawed rooster sperm after addition of resveratrol [36]. ...
Article
The aim of the study was to investigate the effects of different concentrations of resveratrol on head morphology, motility characteristics, oxidative state and in vitro fertility of cooled ram spermatozoa. Pooled semen from three Najdi rams was diluted with Triladyl® having different concentrations of resveratrol, zero (control), 200 μM (45.65 μg/mL) and 400 μM (91.30 μg/mL) resveratrol, then stored at 5 °C for 168 h. The head morphometric, sperm kinematic parameters, Malondialdehyde (MDA), reduced glutathione (GSH), superoxide dismutase (SOD) and in vitro fertilizing capability of ram spermatozoa were evaluated after 24, 72, 120 and 168 h of cooling storage. The total motility (TM) of the sperm with resveratrol at 200 μM and 400 μM was significantly (p ≤ 0.05) higher than that in the control group at 72 and 120 h of cooling storage. On the other hand, the progressive motility (PM) of the sperm with resveratrol at 200 μM and 400 μM was significantly (p ≤ 0.05) higher than that in the control group at 168 h of cooling storage period. After 168 h of cooling storage, significantly higher straightness (STR) was observed in 400 μM group than two other group and in 200 μM group than the control group. Both resveratrol groups had higher linearity (LIN) than control one at 120 and 168 h of cooling storage. The length, width and area of sperm head were lower (P ≤ 0.05) in the control compared to the other treatment groups after 120 and 168 h of storage. There was a significant increase in superoxide dismutase (SOD) concentration in the two resveratrol groups compared with the control one over the seven days of cooling storage and the same result was found in the reduced glutathione (GSH) concentration at 24, 72, and 168 h of storage. There was a significant (p ≤ 0.05) decrease in malondialdehyde (MDA) concentration in the 400 μM resveratrol group than that in two other groups over the seven days of storage period. Cleavage and blastocyst rates following in vitro fertilization were significantly higher (p ≤ 0.05) in 400 μM resveratrol than other group at 72 h for cooling storage period. In conclusion, addition of resveratrol in the extender can protect sperm head morphology, improve kinematic parameters and in vitro fertility, and reduce oxidative stress of ram spermatozoa during liquid storage at 5 °C.
... Hence, adverse climatic conditions in tropical areas lead to oxidative stress, mitochondrial dysfunction which resulted in poor sperm quality. The supplementation of AX @ 2 and 4 µM was found to increase sperm vitality and plasma membrane integrity during the storage period (72 h) (rams' liquid semen) at 4°C, besides reduction in lipid peroxidation and reactive oxygen species (ROS) levels (Fang et al. 2015). Higher sperm motility and Semen analysis: Ejaculates were collected in a graduated centrifuge tube. ...
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This study was conducted to evaluate the effect of astaxanthin (potent herbal antioxidant) supplementation on sperm quality, lipid peroxidation and expression of mitochondrial genes in semen of Karan Fries (Tharparkar × Holstein Friesian) bulls during summer under tropical climatic conditions. Adult healthy bulls (10) were selected and divided equally into 2 groups i.e. control and treatment (supplemented [email protected] 0.25 mg/kg body weight/ day/animal). Ejaculates were collected at weekly interval in early-morning from bulls using artificial-vagina from April to August. Just after collection, semen samples were placed in a water bath (37°C) for semen analysis. Astaxanthin supplementation improved semen quality parameters (volume, motility, concentration, and acrosomeintegrity) over non-supplemented bulls. The major abnormalities were lower in supplemented bulls. Semen malondialdehyde concentration was also lower in treatment than control group. The higher concentration of total antioxidant capacity was observed during July and August in supplemented bulls. Relative expression (mRNA) of succinate dehydrogenase, citrate synthase and mitochondrial transcription factor-A was upregulated in spermatozoa of supplemented bulls than control bulls. Supplementation of astaxanthin to crossbred bulls during summer improved the semen quality by improving the antioxidant activity and modulating the mitocondrial gene expression during the summer season in the tropical climate. Therefore, astaxanthin supplementation could be suggested for improving the semen quality of crossbred bulls during summer season. © 2018 Indian Council of Agricultural Research. All rights reserved.
... Also, in the present study, the addition of 10 and 25 µM canthaxanthin significantly improved the sperm viability. Similarly, it was shown that astaxanthin increases the sperm motility, viability, membrane integrity and reduces the ROS levels (Fang et al., 2015). ...
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Different antioxidants have been introduced to reduce oxidative stress during the cryopreservation. The main goal of this study was to evaluate the effects of canthaxanthin on human sperm parameters during the freeze‐thaw process. This study was performed on 25 normozoospermic semen samples dividing into five groups including 0, 0.1, 1, 10, and 25 µM of canthaxanthin. The prepared spermatozoa were cryopreserved by rapid freezing technique. Sperm motility, viability (eosin‐nigrosin), morphology (Papanicolaou), acrosome reaction (double staining), DNA denaturation (acridine orange), chromatin packaging (aniline blue and toluidine blue), and DNA fragmentation (sperm chromatin dispersion test) were evaluated before freezing and after thawing. All sperm parameters after thawing significantly were decreased compared to before freezing. Twenty‐five micromolar canthaxanthin could significantly improve the progressive and total motility, viability, normal morphology, chromatin packaging, acrosome integrity and DNA denaturation and fragmentation. Ten micromolar canthaxanthin significantly improved total motility, viability, normal morphology, chromatin packaging, acrosome integrity and DNA denaturation and fragmentation. Whereas, in 1 µM group, there were significant differences only in improvement of acrosome integrity, chromatin packaging (toluidine blue) and DNA denaturation and fragmentation. But, in 0.1 µM group, there were no significant differences in any of measured parameters. It seems that canthaxanthin ameliorates detrimental effects of cryopreservation on human sperm parameters.
... In fertilization, sperm provides genetic material to oocyte, and this genetic material was sufficient to affect the quality of embryos. In previous study, sperm quality was improved on long-term storage of semen (Fang et al., 2015) and the modified medium (Dias et al., 2014), motility improvement (Aitken et al., 2012), and inhibition of DNA damage (Lewis et al., 2013). ...
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The aim of present study was to determine the effect of astaxanthin supplementation of the semen extender on post-thaw semen quality and fertility. Semen samples were collected from four fertile rams by using an artificial vagina, pooled and diluted with a Tris–citric acid–fructose–yolk extender, supplemented with AST at 0.5, 1, 2, or 4 μM concentraitons. For the control group, semen was processed in the basic extender without AST. Extended semen was aspirated into straws which were cooled to 5 °C and frozen on liquid nitrogen vapor. After 24 h, the straws were thawed in a water bath for 30 s. The results showed that supplementation of the extender with 2 and 4 μM AST increased the sperm viability and plasma membrane integrity in frozen-thawed semen compared to other groups (P < 0.05), whilst there was no significant difference between extenders containing 0.5 and 1 μM AST. Freezing the sperm in the extenders containing 2 and 4 μM AST led to lower percentages of acrosome abnormalities (27.20% ± 0.54 and 28.15% ±0.54, respectively) and malondialdehyde formation (1.70 ± 0.1 and 1.81 ± 0.2 nmol/ml, respectively ) in comparison to other treatments (P < 0.05). There was no effect of AST on SOD and GSH-PX activities (P > 0.05). Higher fertility rates were obtained when semen extender contained 2 μM (48.33%) and 4 μM (45.76%) AST compared with control group (21.15%). The results showed that, inclusion of AST in frozen ram semen extenders could improve seminal quality and fertility rate.
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Contents 2 Astaxanthin is a member of the carotenoid family well known for its anti-cancer, anti-diabetic, 3 anti-inflammatory, and antioxidant nature. This study was designed to investigate the effects of 4 astaxanthin supplementation of the extender (buffer 2) on post-thaw dog semen quality. Semen from 5 four healthy dogs was collected by digital manipulation twice a week. The ejaculates were pooled, 6 washed, divided into four equal aliquots, diluted with the extender supplemented with different 7 concentrations of astaxanthin (0, 0.5, 1, and 2 µM), and cryopreserved. The results showed that 1 µM 8 astaxanthin was the optimum concentration that led to significantly higher (p < 0.05) post-thaw 9 motility, kinematic parameters, and viability than the other groups. In comparison with the control 10 group, sperm samples supplemented with 1 µM astaxanthin showed significantly higher (p < 0.05) 11 sperm counts with intact membranes (55.7 ± 0.6% vs. 51.3 ± 0.9%), intact acrosome (58.4 ± 0.7% vs. 12 53.5 ±0.6%), active mitochondria (54.9 ± 0.5% vs. 42.6 ± 0.6%), and normal chromatin (67.6 ± 0.9% 13 vs. 61.7 ± 0.6%). Furthermore, astaxanthin-supplemented samples showed significantly lower 14 expression levels (p < 0.05) of pro-apoptotic (BAX), oxidative induced DNA damage repair (OGG1), 15 oxidative stress-related (ROMO1) genes and higher expression levels of anti-apoptotic (BCL2), and 16 sperm acrosome associated (SPACA3) genes compared to the control. Thus, supplementation of 1 µM 17 astaxanthin in semen extender results in improved freeze-thaw sperm quality of the dog. 18
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Prescription of antioxidants might increase the quality of sperm parameters and improve the rate of pregnancy in obese people who suffer from infertility. Therefore, the present study investigated protective effects of vitamin A, E and astaxanthin on sperm parameters and seminiferous tubules epithelium in high-fat diet model. Thirty-six numbers of 3 months old albino Wistar rats were divided to control, high-fat diet and high-fat diet with antioxidants groups. After 12 weeks, levels of LDL-C and HDL-C were detected in the groups. Sperm was obtained from the tail of epididymis and its parameters (count, vitality, motility and morphology) were analyzed. Testes were fixed in 10% formalin and after tissue processing, stained with Hematoxylin and Eosine (H&E) for histological evaluation. Data were analyzed by a one-way ANOVA and p < 0.05 was considered significant. Our results indicated that viability, motility and normal morphology of sperm in high-fat diet (HFD) decreased significantly compared to high-fat diet with antioxidant (HFD + A) and the control groups (p < 0.05). Also spermatogonium and the number of Sertoli cells increased significantly in HFD + A compared to the control (p < 0.05). As it is shown in our study, application of antioxidants decreased serum triglyceride, cholesterol and HDL-C/LDL-C in high-fat diet model and improved the semen parameters. Therefore, it is suggested that the low quality of sperm can be improved in obese men through antioxidant prescription. Finally, it seems that the antioxidants in obese patients with subfertility or infertility is a new and efficient strategy with few side effects.
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The aim of this study was to examine the effect of different doses of vitamin B12 on some physicochemical parameters and antioxidtive enzyme activities in crossbreed rams semen during storage at 5°C. Semen samples were collected from eight crossbreed rams, evaluated and pooled at 33°C. Fresh semen was diluted with a Tris-based extender containing 0, 1, 2 and 3 mg/mL vitamin B12 and was cooled at 5°C. In both genetic group, the extender supplemented with vitamin B12 (1, 2 and 3 mg/mL) led to higher motility percentages than control group. While, the addition of 2 mg/mL vitamin B12 into the semen extenders led to higher viability sperm, in comparison to control group. In Ghezel × Baluchi genotype, the percentage of spermatozoa abnormality was reduced with vitamin B12 when compared with control group. Supplementation with vitamin B12 improved significantly sperm membrane integrity in both genotypes. Addition of vitamin B12 did not cause significant differences in the levels of superoxide dismutase (SOD) and glutathione peroxidase (GPx) when compared with the control group in both genotypes. In the current study, the addition of 2 mg/mL of vitamin B12 (as an antioxidant) to extender had higher SOD activities than the other groups in both genetic groups. In conclusion, vitamin B12 supplementation in semen extender benefit the motility and viability of crossbreed ram sperm.
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The effects of the carotenoids β-carotene and astaxanthin on the peroxidation of liposomes induced by ADP and Fe2+ were examined. Both compounds inhibited production of lipid peroxides, astaxanthin being about 2-fold more effective than β-carotene. The difference in the modes of destruction of the conjugated polyene chain between β-carotene and astaxanthin suggested that the conjugated polyene moiety and terminal ring moieties of the more potent astaxanthin trapped radicals in the membrane and both at the membrane surface and in the membrane, respectively, whereas only the conjugated polyene chain of β-carotene was responsible for radical trapping near the membrane surface and in the interior of the membrane. The efficient antioxidant activity of astaxanthin is suggested to be due to the unique structure of the terminal ring moiety.
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There is evidence that renal transplant recipients have accelerated atherosclerosis manifest by increased cardiovascular morbidity and mortality. The high incidence of atherosclerosis is, in part, related to increased arterial stiffness, vascular dysfunction, elevated oxidative stress and inflammation associated with immunosuppressive therapy. The dietary supplement astaxanthin has shown promise as an antioxidant and anti-inflammatory therapeutic agent in cardiovascular disease. The aim of this trial is to investigate the effects of astaxanthin supplementation on arterial stiffness, oxidative stress and inflammation in renal transplant patients. This is a randomised, placebo controlled clinical trial. A total of 66 renal transplant recipients will be enrolled and allocated to receive either 12 mg/day of astaxanthin or an identical placebo for one-year. Patients will be stratified into four groups according to the type of immunosuppressant therapy they receive: 1) cyclosporine, 2) sirolimus, 3) tacrolimus or 4) prednisolone+/-azathioprine, mycophenolate mofetil or mycophenolate sodium. Primary outcome measures will be changes in 1) arterial stiffness measured by aortic pulse wave velocity (PWV), 2) oxidative stress assessed by plasma isoprostanes and 3) inflammation by plasma pentraxin 3. Secondary outcomes will include changes in vascular function assessed using the brachial artery reactivity (BAR) technique, carotid artery intimal medial thickness (CIMT), augmentation index (AIx), left ventricular afterload and additional measures of oxidative stress and inflammation. Patients will undergo these measures at baseline, six and 12 months. The results of this study will help determine the efficacy of astaxanthin on vascular structure, oxidative stress and inflammation in renal transplant patients. This may lead to a larger intervention trial assessing cardiovascular morbidity and mortality. ACTRN12608000159358.
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There is a lack of information regarding lipid peroxidation and antioxidant capacity in cryopreserved ram semen, and cryopreservation is associated with the production of reactive oxygen species (ROS) which lead to lipid peroxidation (LPO) of sperm membranes, resulting in a loss of motility, viability and fertility of sperm. The aim of this study was to determine the influence of certain additives and their different doses on standard semen parameters, lipid peroxidation and antioxidant activities after the cryopreservation/thawing of ram semen. Ejaculates collected from four Akkaraman rams, a native breed of sheep, were evaluated and pooled at 33 degrees C. Semen samples which were diluted with a Tris-based extender containing additives including trehalose (50, 100mM), taurine (25, 50mM), cysteamine (5, 10mM), and hyaluronan (0.5, 1mg/ml), and an extender containing no additives (control) were cooled to 5 degrees C and frozen in 0.25ml French straws, being stored in liquid nitrogen. Frozen straws were thawed individually at 37 degrees C for 20s in a water bath for evaluation. The use of a Tris-based extender supplemented with 50mM trehalose, 25mM taurine, and 5 and 10mM cysteamine led to higher percentages of post-thaw motility, in comparison to the control group (P<0.01). No significant differences were observed in the percentages of acrosome and total abnormalities, and the hypoosmotic swelling test upon the supplementation of the freezing extender with antioxidants after the thawing of semen. In biochemical assays, the addition of antioxidants did not cause significant differences in levels of malondialdehyde (MDA), glutathione (GSH), and glutathione peroxidase (GSH-Px), after thawing, when compared to groups with no additives. In this study, catalase (CAT) activities were higher in the group that was applied 25mM taurine as an antioxidant, than in all of the other groups (P<0.001). Compared to the controls, antioxidant treatment with 100mM trehalose, 50mM taurine, 5mM cysteamine and 0.5mg/ml hyaluronan, significantly elevated vitamin E (vit E) levels in samples (P<0.001).
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During cryopreservation, oxidative stress exerts physical and chemical changes on sperm functionality. In the present study we investigated the antioxidant effect of rosmarinic acid (RA) on quality and fertilising ability of frozen-thawed boar spermatozoa. Ejaculates collected from mature boar were cryopreserved in lactose-egg yolk buffer supplemented with different concentrations of RA (0 μM, 26.25 μM, 52.5 μM and 105 μM). Motion parameters, acrosome and plasma membrane integrity, lipoperoxidation levels, DNA oxidative damage (8-hydroxy-2-deoxyguanosine base lesion) and in vitro fertilisation ability were evaluated. Total and progressive motility were significantly higher in experimental extenders with RA than in the control (P <0.05) at 0 and 120 min post-thawing. The plasma and acrosomal membrane integrity were improved by supplementation with 105 μM RA (P <0.05). Negative correlation between RA and malondialdehyde (MDA) concentration were determined (P <0.05). After thawing, the percentage of spermatozoa with oxidised DNA did not differ between extenders, however, at 120 and 240 min post-thawing, the samples supplemented with 105 μM RA showed the lowest DNA oxidation rate (P <0.05). The penetration rate was significantly higher on spermatozoa cryopreserved with 105 μM RA (P<0.05). The results suggest that RA provides a protection for boar spermatozoa against oxidative stress during cryopreservation by their antioxidant properties.
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Purpose: The effects of astaxanthin (Ax) on the in vitro development of bovine embryos cultured under heat stress were investigated in combination with the assessment of its cellular accumulation and action on mitochondrial membrane potential (ΔΨm). Methods: Bovine ≥8-cell embryos were collected on day 3 after in vitro fertilization and exposed to single (day 4) or repeated (day 4 and 5) heat stress (10 h/day at 40.5 °C). Ax was added into culture medium under the repeated heat stress and blastocyst development was evaluated. The cellular uptake of Ax in embryos was examined using bright-field and confocal laser-scanning microscopy, and high-performance liquid chromatography. The relationship between Ax and mitochondria localization was assessed using MitoTracker dye. The effects of Ax on ΔΨm were investigated using JC-1 dye. Results: Blastocyst development in the repeated heat stress treatment decreased significantly (P < 0.05) compared with those in single heat stress or normal thermal treatment. The addition of Ax into culture medium did lead to a significant recovery in blastocyst development in the repeated heat-treated group. Ax was detected in cytoplasm of embryos and observed to colocalize with mitochondria. Ax recovered ΔΨm in embryos that was decreased by the heat treatment. Conclusions: Ax ameliorated the heat stress-induced impairment of blastocyst development. Our results suggest that the direct action of Ax on mitochondrial activity via cellular uptake is a mechanism of the ameliorating effects.
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Ecology letters (2011) 14: 891–895 Sperm are particularly prone to oxidative damage because they generate reactive oxygen species (ROS), have a high polyunsaturated fat content and a reduced capacity to repair DNA damage. The dietary compounds vitamin E and beta-carotene are argued to have antioxidant properties that help to counter the damaging effects of excess ROS. Here in, we tested the post-copulatory consequences for male crickets (Teleogryllus oceanicus) of dietary intake of these two candidate antioxidants. During competitive fertilisation trials, vitamin E, but not beta-carotene, singularly enhanced sperm competitiveness. However, the diet combining a high vitamin E dose and beta-carotene produced males with the most competitive ejaculates, possibly due to the known ability of beta-carotene to recycle vitamin E. Our results provide support for the idea that these two common dietary compounds have interactive antioxidant properties in vivo, by affecting the outcomes of male reproductive success under competitive conditions.
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Oxidative stress contributes to defective spermatogenesis leading to male factor infertility. The aim of this study was to review the current literature on the effects of various antioxidants to improve fertilisation and pregnancy rates. The sources of literature were Pubmed and the Cochrane data base. Reviewing the current literature revealed that Carnitines and vitamin C and E have been clearly shown to be effective by many well-conducted studies and may be considered as a first line treatment. The efficacy of antioxidants, such as glutathione, selenium and coenzyme Q10 has been demonstrated by few, but well-performed studies, and may be considered second line treatment. There is, however, a need for further investigation with randomised controlled studies to confirm the efficacy and safety of antioxidant supplementation in the medical treatment of idiopathic male infertility as well as the need to determine the ideal dose of each compound to improve semen parameters, fertilisation rates and pregnancy outcomes.
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Storage of buffalo (Bubalus bubalis) bull semen in the cryopreserved state is discussed in this article. Fertility rate in buffalo following artificial insemination with frozen-thawed semen is reviewed. To better understand the freezability of bubaline spermatozoa, the available data on biochemical components and the activity of specific enzymes of semen/spermatozoa are given. Moreover, the major factors that may influence the post-thaw viability and fertility of buffalo spermatozoa are examined in detail. In addition, suggestions for improvement in cryogenic procedures for buffalo spermatozoa are also given.
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To determine whether there is any association between sperm membrane integrity as determined by the hypo-osmotic swelling test score and unexplained recurrent miscarriage. Prospective observational study. Tertiary referral center for recurrent miscarriage. Semen samples from 20 male partners of women who had had three or more first trimester miscarriages of unexplained etiology and semen samples from 20 prospective semen donors of unknown fertility potential. Sperm density, sperm motility, sperm morphology, and hypoosmotic swelling test score. There was no difference in the median sperm density, the mean sperm motility, or the mean sperm morphology between the two groups. However, the recurrent miscarriage group had a significantly lower hypo-osmotic swelling test score than the control group. The hypo-osmotic swelling test score is significantly lower in samples from men whose partners have had unexplained recurrent spontaneous abortions. With the exception of cytogenetic abnormalities in peripheral blood karyotype, this is the first study to identify a male factor component in recurrent miscarriage.
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Storage of ram semen in liquid and frozen state, the diluents used for both methods, processing, cooling, freezing and thawing of semen are reviewed. Factors influencing the fertility of stored semen and methods used for improvement are discussed, and fertility results of long-term frozen stored ram semen are also given.
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Appearance, pharmacokinetics, and distribution of astaxanthin E/Z and R/S isomers in plasma and lipoprotein fractions were studied in 3 middle-aged male volunteers (37-43 years) after ingestion of a single meal containing a 100 mg dose of astaxanthin. The astaxanthin source consisted of 74% all-E-, 9% 9Z-, 17% 13Z-astaxanthin (3R,3'R-, 3R,3'S; meso-, and 3S,3'S-astaxanthin in a 1:2:1 ratio). The plasma astaxanthin concentration--time curves were measured during 72 hr. Maximum levels of astaxanthin (1.3 +/- 0.1 mg/L) were reached 6.7 +/- 1.2 hr after administration, and the plasma astaxanthin elimination half-life was 21 +/- 11 hr. 13Z-Astaxanthin accumulated selectively, whereas the 3 and 3'R/S astaxanthin distribution was similar to that of the experimental meal. Astaxanthin was present mainly in very low-density lipoproteins containing chylomicrons (VLDL/CM; 36-64% of total astaxanthin), whereas low-density lipoprotein (LDL) and high-density lipoprotein (HDL) contained 29% and 24% of total astaxanthin, respectively. The astaxanthin isomer distribution in plasma, VLDL/CM, LDL, and HDL was not affected by time. The results indicate that a selective process increases the relative proportion of astaxanthin Z-isomers compared to the all-E-astaxanthin during blood uptake and that astaxanthin E/Z isomers have similar pharmacokinetics.
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In this work, molecularly imprinted microspheres (MIMs) were synthesized by aqueous microsuspension polymerization using astaxanthin (3,3'-dihydroxy-beta,beta'-carotene-4,4'-dione) as imprinting molecule. The MIMs obtained were subsequently packed into the stainless steel column and the chromatographic characterization of the column was investigated. The effects of pH and composition of the mobile phase on the retention factor (k') were investigated in detail. The mixture of methanol and dichloromethane (DCM) (8:2, v/v) was used as mobile phase A while the mixture of methanol and water (5:5, v/v) as mobile phase B. The separation of astaxanthin and zeaxanthin (3,3'-dihydroxyl-beta-carotene) was obtained when the concentration of mobile phase B was higher than 30% (v/v) due to their strong lipophilicity. The method developed was successfully applied to separate astaxanthin in the saponified samples of the microalga Haematococcus pluvialis and the yeast Phaffia rhodozyma. The recovery of adding 40 mg astaxanthin to 1.0 g microalgal sample was 95.5% with an R.S.D. (n =5) of 5.3%. The results of determination of astaxanthin in the microalga and the yeast were 3.7% (R.S.D (n = 1.5%, n = 9) and 0.041% (R.S.D n= 7.3%, n = 9), respectively.
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The objectives of this study were (1) to determine the changes in structural, functional and motility parameters of ram-lamb semen stored at two different concentrations at 4 degrees C for 8 days in egg-yolk based extender and (2) to determine the effect of breed of ram-lambs on the changes in structural, functional and motility parameters of ram-lamb semen from different breeds stored at two different concentrations at 4 degrees C for 8 days in egg-yolk based extender. Two different concentrations suitable for laparoscopic and cervical insemination were employed in this experiment. A total of 14 ram-lambs (Polled Dorset-5, Suffolk-5, Katahdin-4) with satisfactory breeding potential were selected. Semen samples were collected by electro-ejaculation. Semen samples were extended to 50 and 200 million sperm per ml with a commercial egg yolk based extender (Triladyl, Minitube of America, Verona, WI, USA) at room temperature and were stored at 4 degrees C. The sperm DNA fragmentation index (DFI), percentages of high mitochondrial membrane potential (hMMP) and plasma membrane integrity (PMI) were assessed using flow cytometry as part of structural and functional parameters on Days 0, 1, 4, 6, and 8. A computer assisted sperm analyser (HTM-IVOS, Version 10.8, Hamilton Thorne Research, Beverly, MA, USA) was used to assess the sperm motility parameters on Days 0, 1, 4, 6, and 8. PROC MIXED procedure was used to determine the effect of days of storage, concentration and breed. The concentration and days of storage significantly affected the sperm structural, functional and motility parameters (P<0.0001). Significant concentration x days of storage interaction was found for all structural and functional parameters. There was a significant concentration x days of storage interaction for average path velocity, curvilinear velocity, straightness and linearity. Overall changes in the sperm structural, functional and sperm motility parameters over the storage period were less dramatic in the 200 x 10(6) ml(-1) concentration when compared to 50 x 10(6) ml(-1) concentration. The hMMP and total progressive motility were influenced by breed. In conclusion, the quality of structural, functional and motility parameters declined as days of storage were increased and the magnitude of changes in the parameters was less dramatic at the higher concentration.
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