Combination of trehalose and low boron in presence of decreased glycerol improves post‐thawed ram sperm parameters: A model study in boron research

Andrology ◽  
2021 ◽  
Author(s):  
Mustafa Numan Bucak ◽  
Nazan Keskin ◽  
Mustafa Bodu ◽  
Bülent Bülbül ◽  
Mesut Kırbaş ◽  
...  
Keyword(s):  
Sperm Parameters ◽  
Model Study ◽  
Ram Sperm

The synergistic effect of trehalose and low concentrations of cryoprotectants can improve post-thaw ram sperm parameters

Cryobiology ◽  
2020 ◽  
Vol 95 ◽  
pp. 157-163 ◽  
Author(s):  
Ali Erdem Öztürk ◽  
Mustafa Bodu ◽  
Mustafa Numan Bucak ◽  
Vahit Ağır ◽  
Ayşe Özcan ◽  
...  
Keyword(s):  
Synergistic Effect ◽  
Sperm Parameters ◽  
Low Concentrations ◽  
Ram Sperm

scholarly journals Positive effects of trehalose and cysteine on ram sperm parameters

2017 ◽  
Vol 62 (No. 5) ◽  
pp. 245-252 ◽  
Author(s):  
S. Gungor ◽  
C. Ozturk ◽  
AD Omur
Keyword(s):  
Sperm Motility ◽  
Sperm Parameters ◽  
Control Group ◽  
Mitochondrial Activity ◽  
Sperm Viability ◽  
Significant Protection ◽  
Positive Effects ◽  
Liquid Storage ◽  
Acrosome Integrity ◽  
Ram Sperm

The aim of this study was to determine the effects of trehalose and cysteine on sperm motility, viability, mitochondrial activity and acrosome integrity during liquid storage of Merino ram semen. Ejaculates were collected using artificial vaginas from five Merino rams, microscopically evaluated and pooled at 37 °C. The pooled semen samples were diluted in a Tris-based extender, including cysteine (2 mM and 4 mM), trehalose (10 mM and 25 mM) and no antioxidant (control). Diluted semen samples were kept in tubes and cooled from 37 to 5 °C in a cold cabinet, and maintained at 5 °C. Cooled samples were evaluated for sperm motility, viability, mitochondrial activity and acrosome integrity at 0, 24, 48, 72 and 96 h. Extender supplemented with trehalose (10 and 25 mM) and cysteine (2 and 4 mM) led to higher motility in comparison to the control at 24, 48, 72 and 96 h of liquid storage (P < 0.05). Trehalose at the doses of 10 mM, 25 mM and 2 mM cysteine led to higher viability between 24–48–72 h and at 96 h of liquid storage (P < 0.05). Further, 4 mM of cysteine improved sperm viability rates at 24 and 48 h of storage compared to the control group (P < 0.05), and resulted in improved acrosome integrity rates compared to the control group at 72 and 96 h of storage (P < 0.05). Extender supplemented with 10 and 25 mM trehalose at 24 and 72 h and 4 mM cysteine at 24 and 96 h of storage led to higher sperm mitochondrial activity rates when compared to the control group (P < 0.05). In conclusion, the findings of this study show that trehalose and cysteine provided significant protection to ram sperm parameters during liquid storage.

Methionine supplementation improves ram sperm parameters during liquid storage at 5°C

2013 ◽  
Vol 13 ◽  
pp. 23
Author(s):  
Kenan Çoyan ◽  
Mustafa Numan Bucak ◽  
Caner Öztürk ◽  
Şükrü Güngör ◽  
Ali Doğan Ömür
Keyword(s):  
Sperm Parameters ◽  
Liquid Storage ◽  
Methionine Supplementation ◽  
Ram Sperm

Effects of Trehalose and Catalase on the Viability and Kinetic Parameters of Cryopreserved Ram Sperm

2018 ◽  
Vol 46 (1) ◽  
pp. 7 ◽  
Author(s):  
Sukru Gungor ◽  
Ayhan Ata ◽  
Muhammed Enes Inanc
Keyword(s):  
Kinetic Parameters ◽  
Sperm Quality ◽  
Sperm Parameters ◽  
Antioxidant Systems ◽  
Control Group ◽  
Positive Effects ◽  
Group 3 ◽  
Group 5 ◽  
Ram Sperm ◽  
Group 1

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, Methods & Results: At the out of breeding season (March-May) seven rams (1-3 years of age) were used in this study. Ejaculates were collected by electro-ejaculator twice a week. Pooled ejaculates were at 37°C, divided into six aliquots, diluted with the Tris based extender containing Trehalose 25 mM (Group-1), Trehalose 50 mM (Group-2), Catalase 200 µg (Group-3), Catalase 400 µg (Group-4), Trehalose 50 mM + Catalase 400 µg (Group-5) and no anti-oxidant (control), respectively, were cooled to 5°C than frozen in 0.25 mL French straws on the nitrogen vapour and stored in liquid nitrogen. The extender supplemented with Group 1 (54.1 ± 1.53; 73.1 ± 4.37), 50 mM (58.3 ± 4.01; 63.1 ± 0.30) and Group 5 (56.6 ± 1.05; 58.3 ± 0.55) resulted in higher subjective motility in comparison to the control (40.0 ± 3.87; 40.5 ± 0.22) group respectively (P < 0.05). Besides, Group 1 (60.16 ± 4.39) and Group 5 (59.60 ± 2.21) led to higher CASA total motility when compared to control (44.40 ± 8.13) group (P < 0.05). Sperm progressive motility was better in Group 1 (20.57 ± 6.90) than the Group 3 (10.63 ± 3.59) [P < 0.05]. Casa kinetic parameters of catalase 200 (Group 3) was higher values than other groups in VCL, VSL, VAP, LIN parameters (P < 0.05). There were no statistically significant differences on the membrane integrity parameter between the groups (P > 0.05).Discussion: Freezing ram sperm is extremely difficult process when compared bull and dog semen. Previous studies showed that antioxidants which were adding into the ram freezing extender gave positive effects solely or combination. In this study similar results were taken at trehalose 25, 50 mM and trehalose 50 mM + catalase 400 µg except 200 and 400 µg catalase groups. These findings supported some researches but lots of them opposite of catalase results. Catalase is found semen and ameliorates the sperm parameters when adding the liquid storage.  Also after diluted and equilibrated catalase groups motilities were better than the control group. During the freezing stage catalase efficiency has been restricted. On the other hand when it combined with the trehalose 50 mM, catalase activity was triggered. Trehalose acts on sperm as non-permanent had a protective action related both osmotic effect and specific interactions with membrane phospholipids. Our data suggest that solely Trehalose 50 mM or combination with Catalase 400 µg can be added to Tris based extender for improving the post-thawed sperm quality in ram semen.

scholarly journals Mini-percoll gradient may be used for frozen-thawed sperm selection in sheep

2019 ◽  
Vol 71 (2) ◽  
pp. 455-463 ◽  
Author(s):  
C.C.S. Olivares ◽  
J.M.G. Souza-Fabjan ◽  
J.F. Fonseca ◽  
H.F.R.A. Saraiva ◽  
L.R. Côrtes ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Centrifugal Force ◽  
Recovery Rate ◽  
Handling Time ◽  
Sperm Parameters ◽  
Sperm Selection ◽  
Percoll Gradient ◽  
Ram Sperm ◽  
First Time ◽  
Motility Parameters

ABSTRACT This study evaluated the effect of increasing centrifugal force and reducing centrifugation time and volume in Percoll protocols on ram sperm parameters. Commercial semen of Santa Inês rams were used and five treatments were performed: traditional Percoll and mini-Percoll (MP) techniques (I- 5000 x g, 5min; II- 2500 x g, 5min; III- 1250 x g, 5min; IV- 700 x g, 10min). At post-thawing (PT) and post-selection protocols (0h), samples were assessed for spermatozoa recovery rate, motility, plasma membrane (PM) integrity, sperm capacitation and morphology and incubated at 37 C for 1, 2 and 3h. The sperm recovery rate averaged 9.1±1.4%, and most motility parameters were similar (P> 0.05) among protocols. VCL (µm/s) was higher (P< 0.05) after MP-II, III and IV (66.1±4.5) than traditional Percoll (46.3±4.9). Capacitation status and PM integrity were similar (P> 0.05) among treatments. For the first time, we have demonstrated the reduction of the gradient volume and centrifugation time associated with an increase on centrifugation force at Percoll can be successfully used for frozen-thawed ram sperm selection. MP may be used instead of traditional Percoll, decreasing costs and semen handling time.

Raffinose and hypotaurine improve the post-thawed Merino ram sperm parameters

Cryobiology ◽  
2013 ◽  
Vol 67 (1) ◽  
pp. 34-39 ◽  
Author(s):  
Mustafa Numan Bucak ◽  
Nazan Keskin ◽  
Mehmet Taşpınar ◽  
Kenan Çoyan ◽  
Nuri Başpınar ◽  
...  
Keyword(s):  
Sperm Parameters ◽  
Ram Sperm

scholarly journals Mini-Percoll Technique Induces Similar Capacitation Features in Domestic Ruminant Frozen-Thawed Spermatozoa Regardless of the Presence of Heparin

2019 ◽  
Vol 47 (1) ◽  
Author(s):  
Vivan Angélico Pereira Alfradique ◽  
Carolina Cerqueira Sarmento Olivares ◽  
Joanna Maria Gonçalvez de Souza-Fabjan ◽  
Jeferson Ferreira Da Fonseca ◽  
Ribrio Ivan Tavares Pereira Batista ◽  
...  
Keyword(s):  
Incubation Medium ◽  
Reproductive Tract ◽  
Female Reproductive Tract ◽  
Sperm Parameters ◽  
Ruminant Species ◽  
Cryopreserved Sperm ◽  
Seminal Plasma Proteins ◽  
Ram Sperm ◽  
Bovine Species

Background: Sperm capacitation is a process consists of a series of functional, biochemical, and biophysical modifications that render the ejaculated sperm competent for oocyte fertilization. Secreted by the female reproductive tract epithelium, heparin promotes capacitation by binding to and removing seminal plasma proteins, which are adsorbed to the sperm PM and would inhibit capacitation. There is substantial evidence that cryopreservation promotes capacitation-like changes in bull, ram and buck sperm. Our general hypotheses were: (a) cryopreserved ram sperm suffer capacitation more quickly than buck and bull sperm under the same conditions; (b) the capacitation status of ruminant cryopreserved sperm is similar whether or not heparin is present after the mini-Percoll technique; and (c) ruminant frozen-thawed sperm selected by mini-Percoll and incubated within media without heparin supplementation is not impaired in terms of capacitation status and sperm agglutination. This study aimed to compare sperm parameters of ovine, caprine, and bovine frozen-thawed sperm after mini-Percoll processing followed by incubation with or without heparin supplementation.Materials, Methods & Results: Commercial semen of all species were used. Sperm samples were selected by mini-Percoll and supplemented (or not) with heparin within an incubation medium for 18 h. Sperm kinematics (CASA system analyzes), capacitation status (CTC staining) and sperm agglutination were evaluated after thawing, mini-Percoll, 1.5 h, 3 h, 6 h and 18 h. In comparison with post-thawing analysis, ovine species demonstrated a reduction (P < 0.05) in most of the sperm motility parameters after mini-Percoll. Conversely, ovine samples presented the highest (P < 0.05) rate of acrosome-reacted cells after mini-Percoll. Heparin supplementation did not affect most of the parameters evaluated (P > 0.05). In caprine and bovine species, a lower (P < 0.05) rate of sperm agglutination was observed in the presence of heparin at 18 h of incubation. In the absence of heparin, ovine samples showed a higher (P < 0.05) agglutination rate compared to the bovine species after long incubation period.Discussion: The present study compared sperm parameters (sperm kinematics, agglutination rate and capacitation status) of ruminant frozen-thawed sperm after mini-Percoll selection followed by in vitro incubation with or without heparin supplementation. In this study, it was observed the same rate of capacitated cells after the sperm selection (min-Percoll) between ruminant species. This indicate that the capacitation process occurs similarly between ruminant species, refuting the first hypothesis of this study. The presence of heparin did not influence the capacitation status of ruminant frozen-thawed sperm after mini-Percoll selection, it demonstrates that the second hypothesis was supported by this study making more economic and practical the use of ruminant frozen-thawed semen. The absence of heparin in the incubation medium did not harmed the capacitation status and sperm agglutination of ruminant frozen-thawed sperm. This supported the third hypothesis of the current study and indicate that the use of mini-Percoll technique regardless the presence of heparin could be a useful alternative for the preparation of ruminant frozen-thawed sperm. In conclusion, the capacitation status of ruminant frozen-thawed sperm is similar whether or not heparin is present after the mini-Percoll technique.

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