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

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.

2018 ◽  
Vol 46 (1) ◽  
pp. 7 ◽  
Author(s):  
Sukru Gungor ◽  
Ayhan Ata ◽  
Muhammed Enes Inanc

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.


2015 ◽  
Vol 55 (5) ◽  
pp. 666 ◽  
Author(s):  
Zh. Bohlool ◽  
M. Mohammadi ◽  
M. Roostaei-Ali Mehr ◽  
N. Ghavi Hossein-Zadeh

This study was conducted to determine the effect of different levels of trehalose and glycerol on ram sperm cryosurvival using lecithin-based diluents. Ejaculates were collected from four rams, pooled after initial evaluation, diluted with Tris-soybean lecithin extender and split into nine equal parts. A total of 0 (T0), 50 (T50) or 100 (T100) mM of trehalose and 3% (G3), 5% (G5) or 7% (G7) of glycerol were added to each part. Sperm motility, viability, plasma membrane integrity and acrosome integrity were evaluated immediately after thawing (0 h), and subsequently after 3 h and 6 h post-thawing incubation at 37°C. Results indicated that there was interaction between trehalose and glycerol on sperm motility. In addition, interaction of trehalose and glycerol with incubation time on sperm motility, viability, plasma membrane integrity and acrosome integrity was not significant (P > 0.05). Sperm motility was greatest in the sperm treated with 100 mM trehalose and 7% glycerol (27%; P < 0.05). The effect of trehalose was significant on viability and plasma membrane integrity of ram spermatozoa (P < 0.05). The main effect of trehalose showed that sperm viability was higher in T100 (47.06%) than T50 (53.96%; P < 0.05). The highest membrane integrity was observed in T100 (47.04%; P < 0.05). Membrane integrity was higher (P < 0.05) in G5 (49.97%) than G3 (41.56%) and there was no difference between G7 (46.86%) and G3 (41.56%; P > 0.05). The best sperm viability and plasma membrane integrity was observed at 0 h (65.75% and 51.58%, respectively). It was concluded that simultaneous use of 7% glycerol and 100 mM trehalose had a desirable effect on motility of ram frozen–thawed sperm.


Author(s):  
I. S. Esua ◽  
U. U. Uno ◽  
U. B. Ekaluo

Background and Aim: Tramadol is a potent analgesic effective in the treatment of mild to severe pains. However, the use of the drug can pose a threat to other organs and systems. Therefore, this study evaluated the effect of graded doses of tramadol on sperm profile of male albino rats. Materials and Methods: Eighteen male rats were divided into three groups (A, B and C) using completely randomized design (CRD) with six rats in each group. Rats in group A served as the control group and were given just food and water while groups B and C were given tramadol at 50 and 100 mg/kg body weight (BW) respectively, daily for the period of 65 days. The treatment was administered via oral gavage and at the end of the treatments, the rats were sacrificed. Immediately after sacrifice, a puncture was made in the epididymis with a sterile pin and examined for semen pH. The epididymes were processed for epididymal sperm motility, viability, count and sperm head abnormality. Results: There was no significant difference in the weight of testes and semen pH. Sperm viability, sperm motility, sperm count and weight of epididymes significantly reduced (p<0.05) in tramadol treated animals when compared with the control. Results also indicated statistically significant (p<0.05) increase in sperm head abnormalities in rats treated with tramadol when compared with the control. Conclusion: The results obtained from this study reveal that tramadol has negative effects on weight of epididymes, sperm count, sperm viability, sperm motility and sperm head abnormalities in male albino rat as mammalian models in a dose dependent manner.


2020 ◽  
Vol 7 ◽  
Author(s):  
Mariana Lucía Bertuzzi ◽  
Edita Yola Torres ◽  
Teodosio Huanca ◽  
Deborah Neild ◽  
María Ignacia Carretero

The use of non-commercial and commercial extenders for cooling alpaca sperm has already been reported, the latter showing certain advantages over the first. The Andromed® (AM) extender was created for use in ruminants and has also been tested in ejaculated and epididymal alpaca sperm. According to the manufacturer, this extender does not need the addition of egg yolk (EY); however, it is known that the addition of EY to some extenders improves the preservation of cooled sperm. The objective of this study therefore was to compare a non-commercial extender (Tris) with the addition of EY vs. the commercial extender AM with and without the addition of EY, for cooling alpaca sperm obtained from diverted deferent ducts. Fifteen pools of deferent duct sperm were formed using samples from two or three different males for each. Each sperm pool was evaluated and then divided into three aliquots that were diluted to a final concentration of 30 × 106 sperm ml-1 (0 h) with either: (1) Tris with 20% EY (T-EY), (2) AM, or (3) AM with 20% EY (AM-EY). Samples were cooled to 5°C and the following sperm parameters were evaluated after 24 and 48 h of storage: motility, viability, membrane function, acrosome integrity, morphology, and chromatin condensation. Motility was also evaluated after 72 h of storage. The samples that best preserved progressive and total sperm motility at the 24 and 48 h evaluation periods were the ones diluted with AM-EY, observing that with this extender these motility patterns decreased significantly after 72 h of storage compared to time 0 h (p &lt; 0.05). A significant decrease (p &lt; 0.05) in total and progressive motility was observed at 48 h for the T-EY and AM extender compared to 0 h. AM was the only extender in which the percentages of viable sperm decreased significantly (p &lt; 0.05) after 48 h of conservation. For the rest of sperm parameters evaluated, no significant differences were observed between any of the extenders at any evaluation time. The Andromed® extender with the addition of 20% EY could be an alternative option for cooling alpaca sperm obtained from deferent ducts.


2000 ◽  
Vol 12 (6) ◽  
pp. 337 ◽  
Author(s):  
H. Lambrechts ◽  
F. E. van Niekerk ◽  
S. W. P. Cloete ◽  
W. A. Coetzer ◽  
G. van der Horst

Microscopically evaluated sperm parameters, as well as computer-aided sperm motility analysis (CASMA), were used to assess sperm quality and the effect of cryopreservation on ram semen obtained from two genetically diverse Merino lines. These lines were divergently selected on maternal ranking values for multiple rearing ability from the same base population since 1986. Replacements in the high (+) line were preferentially the progeny of ewes rearing >1 lamb per joining. Progeny of ewes rearing <1 lamb per joining was preferred as replacements in the low (–) line. Sperm quality, as assessed by percentages of live, abnormal and acrosome-intact spermatozoa as well as by motility, was independent (P≤0.20) of line, time of sampling and their interaction in ejaculated samples obtained from the eight rams used as sires in 1995. Sperm quality of frozen–thawed samples was adversely affected (P≤0.01) by cryopreservation and thawing at 35˚C for 30 s relative to fresh ejaculated samples. No consistent differences between lines were found in epididymal sperm samples obtained from 12 slaughtered rams (6 from each line). The adverse effect (P≤0.05) of cryopreservation and thawing at 35˚C for 30 s on sperm viability and motility was also demonstrated for these samples.


Author(s):  
LI. Jingchun ◽  
LI. Qi ◽  
LI. Yanbug ◽  
WEI Guosheng ◽  
SUN Dongbo

The present study was aimed to investigate the effects of negative pressure applied before storage on the quality and fertilization ability of boar semen. Boar semen samples were collected and pooled, and diluted with Modena solution containing 0.4% (w/v) of bovine serum albumin. Negative pressure was applied for 2–5 min using a vacuum pump with a barometer. The pressure applied were 0 (Control), -0.02 MPa (P200), -0.04 MPa (P400), and -0.08 MPa (P800). The sperm motility, acrosome integrity and sperm fertilizing ability were evaluated. Application of –0.04 MPa improved the sperm motility, acrosome integrity and fertilizing ability, compared with the other groups. The sperm motility and acrosome integrity decreased with increasing storage time in vitro. After 5 days, the sperm motility and acrosome integrity of the P400 group were all higher than those of the other groups (P less than 0.05). The cleavage rate (64.5% ± 2.4%) and blastocyst development rate (33.9% ± 2.8%) for semen stored for 7 days were similar to those of fresh semen. In conclusion, application of –0.04 MPa before liquid storage at 17°C can improve the quality and fertilization ability of boar semen.


Andrologia ◽  
2014 ◽  
Vol 47 (5) ◽  
pp. 545-552 ◽  
Author(s):  
M. N. Bucak ◽  
M. B. Ataman ◽  
N. Başpınar ◽  
O. Uysal ◽  
M. Taşpınar ◽  
...  

2020 ◽  
Vol 89 (1) ◽  
pp. 47-53
Author(s):  
Aiste Iljenkaite ◽  
Sigita Kerziene ◽  
Agila Dauksiene ◽  
Zoja Mikniene ◽  
Henrikas Žilinskas ◽  
...  

The current study was carried out to investigate the protective effects of royal jelly supplementation on sperm motility, viability and pH value during the liquid storage of boar semen at 16 °C and 4 °C, at various periods of time (0, 24, 48, 72 and 96 h). Semen samples were collected from 11 boars, diluted with a long-term extender and supplemented with different concentration of royal jelly (0%, 0.5%, 1% and 2%) at a final concentration of 50 × 106 sperm/ml. In the laboratory, the semen was assessed for sperm morphology, viability (eosin-nigrosin staining), subjective motility and objective sperm motility by sperm class analyzer. In total, 396 tests for sperm viability and motility were performed. The longer storage time and the lower incubation temperature showed lower sperm motility and viability results. The results showed that royal jelly supplementation at 1% concentrations protected the functionality of the sperm plasma membrane during the liquid storage time of 96 h at 16 °C. Sperm subjective and objective motility results in samples stored at 4 °C decreased with higher royal jelly concentrations and longer storage time, and differ significantly from the results in samples stored at 16 °C (P < 0.05). Our data showed that royal jelly supplementation at lower concentrations can improve boar semen motility and viability parameters during liquid storage at 16 °C for 96 h.


2020 ◽  
Vol 41 (1) ◽  
pp. 167 ◽  
Author(s):  
Breno Fernandes Barreto Sampaio ◽  
Bruno Gomes Nogueira ◽  
Maria Inês Lenz Souza ◽  
Eliane Vianna da Costa-e-Silva ◽  
Carmem Estefânia Serra Neto Zúccari

Plasma membrane composition has impact on phase transition from liquid crystal to gel state of cooled sperm cell. The incorporation of polyunsaturated fatty acids increases its fluidity and can contribute to sperm motility. The aim of this study was to compare the effect of adding docosahexaenoic acid (DHA) and ?-tocopherol (?-Toh) to the cooling extender, singly or combined, to the equine sperm parameters, submitted to cooling, up to 72 hours. Two ejaculates of ten stallions collected with artificial vagina were used, and evaluated for motility, plasma membrane integrity, chromatin fragmentation, mitochondrial activity and lipid peroxidation, according to the following treatments: C; DHA; ?-Toh; DHA/?-Toh; EtOH 100: and EtOH 140 (corresponding to control; 10 ng mL-1 of DHA; 2 mM of ?-Toh; : 10 ng mL-1 of DHA + 2 mM of ?-Toh; 100 µL of ethanol and 140 µL of ethanol respectively). DHA treatment showed higher motility (68.2 ± 12.3; p < 0.05) when compared to control (62.1 ± 16.2), DHA/?-Toh (61.3 ± 12.7) and EtOH (58.1 ± 8.6) groups. In lipid peroxidation assay, the control group showed 2,506.2 ± 796.4 ng of MDA 108 spermatozoa-1, being significantly higher (p < 0.05) than the groups treated with DHA (2,036.0 ± 687.0), ?-Toh (1,890.8 ± 749.5) and DHA/?-Toh (1,821.1 ± 627.2). In conclusion, ?-Toh was effective in diminishing lipid peroxidation of equine sperm subjected to cooling, and DHA improved sperm motility and, in spite of being a polyunsaturated fatty acid with high susceptibility to peroxidation, reduced lipid peroxidation.


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