Effect of the Breed on the Activity of the Antioxidant Enzymes – Sod and Cat in Ram Sperm, Before and After Cryopreservation (original) (raw)
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Cryobiology, 2017
Ram sperm are subjected to extreme oxidative stress during their preservation at-196°C resulting in reduced quality at post thaw. Therefore, the main objective of this study was to evaluate the effect of antioxidants taurine, quercetin and reduced glutathione on the post thaw quality of crossbred ram sperm. A total of twenty four ejaculates from six crossbred rams were collected and extended with tris-based extender with no antioxidant (Control), with taurine (40 mM), quercetin (5µg/ml) and reduced glutathione (5 mM). The post thaw sperm quality was determined by percent sperm motility, live sperm count, intact acrosome and hypoosmotic swelling test (HOST) reacted spermatozoa and lipid peroxidation was measured in terms of malondialdehyde (MDA) level both in seminal plasma and sperm cell. At post thaw, percent sperm motility and live sperm count were significantly (p<0.05) higher for taurine than control and reduced glutathione but did not differ significantly (p>0.05) from quercetin. The percent HOST reacted spermatozoa were significantly higher for taurine than control, quercetin and reduced glutathione. Seminal plasma MDA level was significantly (p<0.05) lower for taurine than control and non-significantly lower than
2009
Freezing of the semen in liquid nitrogen is the only method for long term storage of fertile spermatozoa. The procedure involves different steps which are harmful to spermatozoa and consequently reduce their quality and fertility. In many animal species the addition of antioxidants to semen before freezing was found to have positive effect on the quality and fertility of frozen/thawed (F/T) spermatozoa. As in rams there are contradictory results in the literature concerning the influence of specific antioxidants on sperm quality of F/T spermatozoa, the present study was performed to evaluate the effects of two different antioxidants on post thaw motility, viability and DNA integrity of F/T ram spermatozoa. Ejaculates from six crossbreed rams were frozen according to the standard procedure after two step dilution with modified Tris-egg yolk extender. The second extender, which contained 14% of glycerol, was added to the semen at 5°C. Ejaculates were divided into three parts and exten...
Theriogenology, 2012
The aim of the present study was to determine the influence of chicken semen cryopreservation on sperm parameters, lipid peroxidation and antioxidant enzymes activities. Pooled semen from 10 Black Minorca roosters was used in the study. Semen samples were subjected to cryopreservation using the "pellet" method and dimethylacetamide (DMA) as a cryoprotectant. In the fresh and the frozen-thawed semen sperm membrane integrity (SYBR-14/propidium iodide (PI)), acrosomal damage (PNA-Alexa Fluor ® 488) and mitochondrial activity (Rhodamine 123) were assessed using flow cytometry. Malondialdehyde (MDA) concentration, catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities were determined in sperm cells and seminal plasma by spectrophotometry. All sperm characteristics evaluated using flow cytometry were affected by cryopreservation. After freezing-thawing, there was significant (P Ͻ 0.01) reduction in sperm membrane integrity, sperm acrosome integrity and mitochondrial activity. Following cryopreservation, MDA concentration significantly increased in chicken seminal plasma and spermatozoa (P Ͻ 0.01, P Ͻ 0.05). The CAT activity in seminal plasma significantly decreased (P Ͻ 0.05), while intracellular activity of this enzyme did not significantly change in frozen-thawed semen. In seminal plasma of frozen-thawed semen the significant increase (P Ͻ 0.01) in GPx activity was detected. Whereas GPx activity in spermatozoa remained statistically unchanged after thawing. The SOD activity significantly increased (P Ͻ 0.01) in cryopreserved seminal plasma with simultaneous decrease (P Ͻ 0.01) of its activity in cells. In conclusion, this is probably the first report describing the level of antioxidant enzymes in frozen-thawed avian semen. The present study showed that the activity of CAT, GPx and SOD in chicken semen was affected by cryopreservation, what increased the intensity of lipid peroxidation (LPO). Catalase appeared to play an important role in the sperm antioxidant defense strategy at cryopreservation since, opposite to SOD and GPx, its content was clearly reduced by the cryopreservation process. Change in the antioxidant defense status of the chicken spermatozoa and surrounding seminal plasma might affect the semen quality and sperm fertilizing ability.
Impact of Seminal Plasma Antioxidants on Donkey Sperm Cryotolerance
Antioxidants, 2022
This study investigated whether the activities of the antioxidant components of donkey seminal plasma (SP)—both enzymatic (superoxide dismutase (SOD), catalase-like (CAT), glutathione peroxidase-like (GPX), and paraoxonase type 1 (PON1)) and non-enzymatic (measured in terms of total thiol, copper-reducing antioxidant capacity (CUPRAC), ferric-reducing ability of plasma (FRAP), and Trolox equivalent antioxidant capacity (TEAC))—and oxidative stress index (OSI) are related to sperm cryotolerance. For this purpose, 15 ejaculates from jackasses (one per individual) were collected and split into two aliquots. The first one was used for measuring the activities levels of enzymatic and non-enzymatic antioxidants and OSI in SP, whereas the other aliquot was cryopreserved. Before cryopreservation, sperm quality parameters (concentration, motility, and viability) were evaluated. After thawing, sperm motility, plasma membrane integrity, lipid disorder, mitochondrial membrane potential, reactiv...
Small Ruminant Research, 2010
The aim of this study was to determine the effects of the antioxidants curcumin, inositol and carnitine on microscopic seminal parameters, lipid peroxidation (LPO) and the antioxidant activities of sperm, following the freeze-thawing of Angora goat semen. Ejaculates were collected via artificial vagina from three Angora goats and microscopically evaluated and pooled at 37 • C. The pooled semen samples were diluted in a Tris-based extender, including curcumin (2.5, 5 or 10 mM), inositol (2.5, 5 or 10 mM), carnitine (2.5, 5 or 10 mM) and no antioxidant (control). The diluted semen was slowly (at a rate of 0.2-0.3 • C/min) cooled to 5 • C and then cryopreserved in 0.25 mL French straws. Frozen straws were thawed individually at 37 • C for 20 s in a water bath, for microscopic sperm evaluation. The freezing extender supplemented with 2.5 mM curcumin led to higher percentage of computerassisted semen analyzer (CASA) sperm motility (65 ± 3%), when compared to the control, inositol and the 10 mM carnitine (P < 0.01) groups, following the freeze-thawing process. The addition of antioxidants did not provide any significant effect on the percentages of post-thaw subjective analyses and CASA progressive motilities, as well as sperm motility characteristics (VAP, VSL, LIN and ALH), compared to the controls. Freezing extenders with antioxidants at three different doses led to lower percentages of acrosome and total sperm abnormalities, when compared to the controls (P < 0.001). However, the addition of 5 mM inositol did not induce any difference in total sperm abnormalities, when compared to the controls. The antioxidants also did not show any effectiveness in the elimination of malondialdehyde (MDA) formation and the maintenance of glutathione peroxidase (GSH-PX) activity, when compared to the controls. Superoxide dismutase (SOD) activity was found to be higher in the presence of curcumin at all three dose levels and carnitine at 5 mM, compared to the other groups. Glutathione (GSH) concentration was demonstrated to be maintained at a higher level with the addition of inositol, compared to the other groups. However, these differences in SOD and GSH levels were not significant, compared to the controls. All the antioxidants at all three dose levels resulted in a better protection of the sperm morphology (except for 5 mM inositol with respect to the total sperm abnormalities), compared to the control samples. According to CASA, the best post-thawing sperm motility M.N. Bucak et al. / Small Ruminant Research 89 (2010) 24-30 25 rate was recorded when the freezing extender was supplemented with 2.5 mM curcumin. Further studies are required to obtain more conclusive results regarding the characterization of microscopic and oxidative stress parameters in cryopreserved goat sperm, using the different antioxidants.
Effects of Trehalose and Catalase on the Viability and Kinetic Parameters of Cryopreserved Ram Sperm
Acta Scientiae Veterinariae, 2018
Background: Most part of ram spermatozoa membrane has unsaturated fatty acids (phospholipids). Membrane structure of cells is composed of double ordered phospholipid layers adorned with mosaic-like protein, glycoprotein and glycolipids. Sperm freezing protocols could be negatively affected on ram sperm motility, viability and acrosome integrity during cryopreservation. For these reasons, researchers were designed their topics has led to the search for effective antioxidant systems against peroxidative damage and spermatozoon dysfunction. There are three protective enzymatic systems against reactive oxygen species (ROS) damage in sperm. These include superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase / reductase cycles Catalase (CAT) is a hemo-protein in the enzyme tetramer structure. The aim of this study was to investigate the effects of trehalose, catalase and their combinations on ram sperm parameters after the cryopreservation/thawing process.Materials, Metho...
Seminal Plasma Antioxidants Are Related to Sperm Cryotolerance in the Horse
Antioxidants
The objective of this study was to determine the relationship of enzymatic (superoxide dismutase, SOD; glutathione peroxidase, GPX; catalase, CAT; and paraoxonase type 1, PON1) and non-enzymatic antioxidants (measured in terms of: Trolox equivalent antioxidant capacity, TEAC; cupric-reducing antioxidant capacity, CUPRAC; and ferric-reducing ability of plasma, FRAP), as well as the oxidative stress index (OSI) in seminal plasma (SP) with the resilience of horse sperm to freeze-thawing. Twenty-one ejaculates (one per individual) were collected and split into two aliquots: the first was used to harvest the SP and assess the activity levels of antioxidants and the OSI, and the second one was cryopreserved. The following post-thaw sperm quality parameters were evaluated: sperm motility, plasma membrane and acrosome integrity, mitochondrial membrane potential, intracellular levels of reactive oxygen species (ROS), and plasma membrane lipid disorder. Based on post-thaw total motility (TM) ...
Seasonal variations in antioxidant enzyme activity in ram seminal plasma
Theriogenology, 2007
Seminal oxidative stress status is emerging as a significant prognostic tool in assisted reproductive technology. A dynamic interplay between pro-and anti-antioxidant substances in the ejaculate is essential. In this study, we determined seasonal changes in the activity of the antioxidant enzyme defense system comprising superoxide dismutase (SOD), glutathione reductase (GR), glutathione peroxidase (GPx) and catalase (CAT) in seminal plasma (SP) of mature Rasa Aragonesa rams. This breed corresponds to a local Spanish genotype with a short seasonal anoestrus between May and July. In addition, the activity of these enzymes was measured in protein fractions isolated from ram SP by exclusion chromatography.
Influence of the Cryopreservation on the Vitality of the Sperm of the Different Breeds of Rams
2020
The aim of the study was to determine the effect of cryopreservation on the vitality of sperm from dif-ferent breeds of rams. For the purpose of the study were used 15 healthy rams from three breeds – Ile-de-France, Lacaune, Synthetic Population of Bulgarian Milk (SPBM), during their insemination campaign. Sperm viability was determined by a Computer-assisted sperm analysis (CASA) after staining with a kit containing nigrosin-eosin (NE) solution. After cryopreservation, the highest percentage of vital sperm were found in the Ile-de-France breed. A significance difference was found between the Ile-de-France and SPBM breeds (P˂0.001) and between Ile-de-France and Lacaune (P˂0.01), regarding the studied indicator