Flow cytometric and microscopic evaluation of post-thawed ram semen cryopreserved in chemically defined home-made or commercial extenders

Emamverdi, Mojtaba; Zhandi, Mahdi; Shahneh, Ahmad Zare; Sharafi, Mohsen; Akhlaghi, A.; Motlagh, Mahdi Khodaei; Dadkhah, Fatemeh; Davachi, Navid Dadashpour

doi:10.1071/an13215

Cited by 26 publications

(11 citation statements)

References 44 publications

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“…that include egg yolk serve to prevent cold shock and provide membrane stabilization during the freeze-thawing process [46]. The stabilization of sperm membranes has an important role in improvement of post thawed sperm quality [47,48].…”

Section: Effect Of Extenders and Insemination Protocol On The Fertilimentioning

confidence: 99%

Effect of Extenders and Insemination Protocol on the Fertilizing Capacity of Cryopreserved Arabian Horse Semen

Waheed¹,

Al-Khaldi²,

Pratap³

2016

SOJVS

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SOJ Veterinary SciencesOpen Access Research Article modifications have been proposed for the freezing process [6]. It has been estimated that only 30-40% of stallions produce semen suitable for cryopreservation [7]. Moreover, consistent variation of sperm freezability has been observed from breed to breed [8,9]. At present, there is no standard protocol for stallion semen cryopreservation able to achieve consistent and acceptable post thaw sperm fertilizing capacity with every ejaculate [10]. That lack of standard can be attributed to many factors, including damage to the sperm membrane that may occur during the process of freezing or thawing [11]. In addition, the cryopreservation extender has an important influence on post thaw sperm quality [11].Many diluents have been used in different animal species, with good results in post thaw sperm motility, but they have not been used with the semen of Arabian horses. Several diluents have been used in stallion semen, with good results such as Kenney's diluent, cream-gel, skim milk diluent, glycine egg yolk, sugar-based extender and INRA 96 extender [12,13]. The use of Computer-Assisted Semen Analysis (CASA), which facilitates objective measurement of several parameters of sperm motility, offers a more reliable and repeatable means of assessing sperm motility than does examination by eye [14]. The CASA system is therefore able to determine a series of variables, including number of moving spermatozoa, Curvilinear Velocity (VCL), Linear Velocity (VSL), Linear Coefficient (LIN), straightness coefficient (STR) and frequency of head displacement (BCF) [15].Success with frozen semen requires attention to detail and a basic understanding of the techniques involved in the proper handling, thawing, and insemination of frozen semen [7]. Frozen stallion semen is now used more extensively by minimizing freeze thaw damage to spermatozoa, and indeed, more stallions might be used and pregnancy rates might be increased. Many steps constitute interactive participation in the success of the cryopreservation process, including semen collection, dilution, cooling, freezing, and thawing [12]. AbstractForty-eight ejaculates were obtained from four Arabian stallions to study the impact of three extenders (INRA Freeze, TRIS-egg yolk and E-Z Mixin) on the fertilizing capacity of cryopreserved stallions' semen. Ejaculates were cryopreserved in 0.5-ml plastic straws with a dilution rate of 1:1 and 1:2. Frozen straws were thawed at either 37°C for 30 sec or 70°C for 7sec and used for AI of eighty Arabian mares via different protocols. Results revealed that percentages of motility; live sperm; and abnormalities were significantly (P < 0.01) better in the E-Z Mixin at 5°C with dilution rates 1:1 and 1:2. Motility characteristics such as path velocity (VAP, μm/s), straightline velocity (VSL,μm/s), Point-to-Point Velocity (VCL, μm/s) and lateral head displacement (ALH, μm) were significantly (P < 0.01) the best by using E-Z Mixin at rates of dilution 1:1 and 1:2. Conception rate was significantly (...

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Section: Effect Of Extenders and Insemination Protocol On The Fertilimentioning

confidence: 99%

Effect of Extenders and Insemination Protocol on the Fertilizing Capacity of Cryopreserved Arabian Horse Semen

Waheed¹,

Al-Khaldi²,

Pratap³

2016

SOJVS

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“…Samples were washed (500 g for 5min) and pellet were resuspended in 250 μL PBS buffer. Then, 10 μL of PI solution (1 mg/mL) was added to the sample and assessed by flowcytometry to determine the number of live spermatozoa with active mitochondria (positive for R123 and negative for PI) [35].…”

Section: Mitochondrial Statusmentioning

confidence: 99%

Improvement of post-thawed sperm quality and fertility of Arian rooster by oral administration of d-aspartic acid

et al. 2017

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This study was conducted to investigate the effect of d-Aspartic acid (D-Asp) on post-thawed sperm quality, fertility and hatchability outcomes in male broiler breeders. Twenty 55-week-old roosters were selected and equally split into four groups (n = 5 rooster/group). Different daily D-Asp doses including 0 (D-0), 100 (D-100), 200 (D-200) or 300 (D-300) mg/kg BW were capsulated and individually administered for 12 weeks to roosters in each group. Semen samples were weekly collected from 7th to 12th week of experiment. Sperm quality from 7th to 11th week was evaluated in both fresh (total and forward motility and plasma membrane functionality) and post-thawed (total and forward motility, plasma membrane functionality, apoptosis status and mitochondrial activity) conditions. Also, collected semen samples on the 12th week were frozen and artificially inseminated to evaluate fertility and hatchability. The results from fresh condition showed that total and forward motility and plasma membrane functionality were significantly higher in D-200 compared to other groups. Also, interaction effect of time and treatment was not significant for all assessed parameters in fresh condition. In post-thawed condition, D-200 showed significantly higher total and forward motility, fertility and hatchability compared to other groups. The higher value for plasma membrane functionality and mitochondrial activity was observed in D-200 compared to D-0 and D300 groups. However, the percentage of live, early apoptotic and dead spermatozoa were not significantly affected by applied treatment in the current study. No significant difference for time and treat interaction effect was observed for all assessed parameters except forward motility. In conclusion, it seems that D-Asp administration could improve fresh and post-thawed sperm quality and post-thawed sperm fertility in male broiler breeders.

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“…Semen cryopreservation has allowed specific opportunities for conservation of genetic resources through sperm banks, guarantee of a constant commercial supply of semen, and collaboration in breed improvement programs by artificial insemination (Bucak et al, 2007;Masoudi et al, 2017). Semen cryopreservation may lead to oxidative, chemical and physical damages on sperm membrane leading to reduction of sperm viability and fertility (Evans et al, 1987;Watson, 2000;Emamverdi et al, 2014;Najafi et al, 2014a;Sarıözkan et al, 2015). Freezing process mostly leads to loss of motility, acrosomal and plasma membrane functionality of spermatozoa (Tuncer et al, 2011;Najafi et al, 2013;Shahverdi et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

Alpha-tocopherol improves frozen-thawed sperm quality by reducing hydrogen peroxide during cryopreservation of bull semen

Motemani

Chamani

Sharafi

et al. 2017

Span. j. agric. res.

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This study was conducted to investigate the effects of different levels of α-tocopherol in cryopreservation media on the bull sperm quality after thawing. Semen samples were collected from six Holstein bulls using artificial vagina twice a week. Semen samples were pooled to eliminate individual differences and then divided into four equal parts for freezing with extenders containing different concentrations of α-tocopherol according to experimental groups as follows: 0 (control), 1.2 mM (E1), 2.4 mM (E2) and 4.8 mM (E4). Motion characteristics, viability, plasma membrane functionality, lipid peroxidation and H 2 O 2 status were determined after thawing. Results showed that malondialdehyde (MDA) concentration was significantly lower in E4 (6.1±0.6 nmol/mL) than E1 and E2 extenders (8.1±0.6 nmol/mL and 8±0.6 nmol/mL, respectively). Also, the lowest significant concentration of H 2 O 2 was observed in E4 (3.2±0.13 nmol/mL) compared to E1 (4 ±0.1 nmol/mL), E2 (5.3± 0.1 nmol/mL) and control (6.7±0.1 nmol/mL). Moreover, E2 and E4 produced the highest significant motility (74.2±1.6%, 75.9±1.6%), viability (78.2±1.8%, 76.1±1.8%) and membrane functionality (73±1.6%, 70.5±1.6%) compared to other groups. It can be concluded that α-tocopherol at the concentration of 4.8 mM can be an efficient antioxidant additive in Bioxcell extender for cryopreservation of bull semen.Additional key words: antioxidant; bioxcell; lipid peroxidation; sperm; H 2 O 2 .Correspondence should be addressed to Mohammad Chamani: m.chamani@srbiau.ac.ir Abbreviations used: LPO (lipid peroxidation); MDA (malondialdehyde); ROS (reactive oxygen species).Funding: The authors received no specific funding for this work.

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Flow cytometric and microscopic evaluation of post-thawed ram semen cryopreserved in chemically defined home-made or commercial extenders

Cited by 26 publications

References 44 publications

Effect of Extenders and Insemination Protocol on the Fertilizing Capacity of Cryopreserved Arabian Horse Semen

Effect of Extenders and Insemination Protocol on the Fertilizing Capacity of Cryopreserved Arabian Horse Semen

Improvement of post-thawed sperm quality and fertility of Arian rooster by oral administration of d-aspartic acid

Alpha-tocopherol improves frozen-thawed sperm quality by reducing hydrogen peroxide during cryopreservation of bull semen

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