scholarly journals Effect of Centrifugation on Motility, Sperm Capacitation and Acrosome Reaction in Soy Bean and Avocado Seed Milk Extenders of Cryopreserved Goat Spermatozoa

2017 ◽  
Vol 50 (1) ◽  
pp. 13-18
Author(s):  
James Ola Daramola
Keyword(s):  
Sperm Motility ◽  
Seminal Plasma ◽  
Acrosome Reaction ◽  
Sperm Quality ◽  
West African ◽  
West African Dwarf ◽  
Artificial Vagina ◽  
Avocado Seed

Abstract Removal of seminal plasma by centrifugation (0 centrifugation, 1 centrifugation, 2 centrifugations, 3 centrifugations) and preservation in two different tris-extenders viz., avocado seed milk (ASM) and soy bean milk (SBM) based extenders were studied for their ability to support motility, in vitro capacitation and acrosome reaction of spermatozoa obtained from West African Dwarf (WAD) goat bucks during cryopreservation. Semen samples collected with the aid of artificial vagina were centrifuged for one, two and three times. The centrifuged samples were diluted with the two tris-extenders each containing 20 mL of avocado seed milk and soybean milk and cryopreserved for 30 days. The results showed higher (P < 0.05) sperm motility (P < 0.05) with increased centrifugation times. Spermatozoa that were centrifuged had higher (P < 0.05) percentage of acrosome reaction and capacitation with increased centrifugation times compared to the control. Optimal improvement in these parameters was obtained with increased centrifugation times. The findings revealed that removal of seminal plasma by centrifugation improved sperm quality of WAD goat bucks during cryopreservation and optimum improvement was achieved consistently with 3 centrifugations.

scholarly journals IMPACT OF AMPK ACTIVATOR METFORMIN ON SPERM QUALITY

2018 ◽  
Vol 15 (4) ◽  
pp. 605-615
Author(s):  
Nguyen Thi Mong Diep ◽  
Elisabeth Blesbois
Keyword(s):  
Sperm Motility ◽  
Acrosome Reaction ◽  
Positive Impact ◽  
Sperm Quality ◽  
Fluorescent Intensity ◽  
Freeze Thaw ◽  
The Media ◽  
Cryopreserved Sperm ◽  
Amp Activated Protein Kinase

Semen cryopreservation allows crucial management of animal genetic diversity. However, the freeze-thaw process causes biochemical and physical alterations, impairing sperm energy-dependent functions. Currently, many chemicals are added to the media to enhance frozen-thawed sperm quality during artificial insemination. The aims of this study was to determine the effects of Metformin (Metf) on fresh chicken sperm motility and ability to perform acrosome reaction, and to evaluate Metf’s effects on the functions of cryopreserved sperm. Chicken semen was diluted and incubated at 35°C in media supplemented with or without different doses of 5’-AMP-Activated Protein Kinase (AMPK) activator, Metf (0,5 to 5 mM). We then looked for the concentration improving the most sperm quality to use it in the cryopreservation media used for chicken sperm. Our results show that 1 mM Metf is the concentration giving the best results regarding sperm quality. AMPKα phosphorylation, viability, acrosome reaction ability (AR), and various motility parameters, were negatively affected by the freeze-thaw process, and that Metf partially restored them. Sperm quality improved (mean increased by 23% for motility, by 10% for viability) as well as AMPKα phosphorylation (mean increased by 30%). Moreover, fluorescent intensity levels of phospho-AMPK were also stronger with Metf than in the control. These results show that the presence of Metf in fresh semen has a positive impact on the quality of sperm and helps reducing the gradual decline in sperm motility caused by cryopreservation by partially restoring several essential sperm functions, and thus leads to a better overall quality of cryopreserved sperm.

Freezability of Andalusian donkey (Equus asinus) spermatozoa: effect of extenders and permeating cryoprotectants

2016 ◽  
Vol 28 (12) ◽  
pp. 1990 ◽  
Author(s):  
D. Acha ◽  
M. Hidalgo ◽  
I. Ortiz ◽  
M. J. Gálvez ◽  
J. J. Carrasco ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Equus Asinus ◽  
Sperm Membrane ◽  
Freeze Test ◽  
Pooled Samples

The aim of this study was to compare the effect of two semen extenders and four permeating cryoprotectants on post-thaw sperm quality of Andalusian donkeys. First, 32 ejaculates were pooled, split and frozen in either Gent B or INRA 96 with egg yolk and glycerol. Second, 12 pooled semen samples were simultaneously frozen in Gent B (glycerol) or Gent A containing ethylene glycol (EG; 1 or 1.5%) or dimethyl sulfoxide (DMSO; 1.5 or 2%). Finally, nine pooled samples were simultaneously cryopreserved in Gent A containing 1% EG (as control), dimethylformamide (DMFA; 1 or 2.5%) or a combination of 1% EG and 1.5% DMFA. Gent B yielded a higher (P < 0.01) post-thaw sperm motility than modified INRA96. EG 1% increased the sperm membrane integrity (P < 0.001), whereas DMSO affected sperm motility and membrane integrity (P < 0.001). DMFA 2.5% yielded higher (P < 0.001) values for sperm motility and membrane integrity. We concluded that Gent B improves in vitro post-thaw sperm quality of donkey spermatozoa, but the replacement of glycerol with 1% EG or 2.5% DMFA increased sperm protection against cryodamage. The use of DMSO for freezing donkey semen was unsuccessful and a toxic effect is suspected. These extenders should be included in the pre-freeze test for each donkey.

scholarly journals Glutathione supplementation in semen extender improves rabbit sperm attributes during refrigeration

2021 ◽  
Vol 29 (2) ◽  
pp. 81
Author(s):  
Ejaz Ahmad ◽  
Zahid Naseer ◽  
Melih Aksoy
Keyword(s):  
Sperm Motility ◽  
Acrosome Reaction ◽  
Reaction Rate ◽  
Sperm Quality ◽  
Final Concentration ◽  
Sperm Viability ◽  
Acrosome Integrity ◽  
Semen Extender ◽  
Sperm Motion ◽  
Artificial Vagina

In the present study, we evaluated the sustaining effect of various glutathione (GSH) concentrations in extender on rabbit sperm attributes during storage at 5°C for 24 h. Semen was collected from regular donor rabbit bucks using an artificial vagina and initially evaluated for sperm quality. The qualifying ejaculates were diluted with one of the extenders having 0, 1, 2, 4 or 8 mM GSH, to achieve a final concentration of 1×108 sperm/mL. The extended samples were stored at 5°C for 24 h. Sperm motility, motion kinetics, acrosome integrity and viability were assessed after 3, 6, 12 and 24 h of storage. The results showed that total sperm motility and sperm motion kinetics (oscillation index of the sperm, straightness index and beat cross frequency) were influenced (<em>P</em>&lt;0.05) by glutathione dose and refrigeration time. An interaction of (<em>P</em>&lt;0.05) GSH concentrations and refrigeration time was observed for sperm viability and acrosome reaction rate. In conclusion, the 4 mM GSH supplemented extender’s protective influence was remarkable to maintainrabbit sperm quality for 24 h 5°C.

scholarly journals Relationship of extender and packaging system an the length of preservation and the quality of chilled semen of Boer goat

2016 ◽  
Vol 21 (1) ◽  
pp. 49
Author(s):  
Arie Febretrisiana ◽  
. Anwar ◽  
Simon Sinulingga
Keyword(s):  
Sperm Motility ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Packaging System ◽  
Boer Goat ◽  
Sperm Membrane ◽  
Relationship Of ◽  
Artificial Vagina

<p class="abstrak2">The aim of this research was to compare the effectiveness of different extender (either Triladyl or Tris Egg Yolk extender) and different packaging method (pool and straw) of chilled semen an the length of preservation and the quality of chilled semen of Boer goat. Semen was collected using an artificial vagina from 3 two years old Boer bucks with body weight of 50-55 kg. It was evaluated under a microscope, then each was diluted either in Tris egg yolk extender (TEY) or Triladyl. Those diluted sperms were then packed either in pool or straw and preserved at 5⁰C refrigerator. Sperm motility, viability and membrane integrity of each group were evaluated every 24 h for up to 5 days. Results showed that sperm motility in Triladyl of  pool packaging system up to 3 days was higher than straw packaging system or TEY in pool or straw packaging system which were 45.8%, 26.1%, 32.1% and 9.1%, respectively (P&lt;0.05). Percentage of sperm membrane integrity showed the same pattern to Triladyl both in pool and straw packaging system which was higher than TEY group (75.2% and 77,2%; P&lt;0.05). Sperm viability in Triladyl both in pool or straw packaging system decreased (P&lt;0.05) after 3 days of preservation (77.1% and 76.2%) but TEY significanly decreased after 4 days of preservation either in pool or straw packaging system (73.2% and 58.0%; P&lt;0.05). It was concluded that sperm quality decreased with increasing of the length of preservation while Triladyl extender in pool packaging system showed the best quality.</p><strong>Key Words: </strong>Chilled Semen, Boer, Triladyl, Tris Egg Yolk, Straw

scholarly journals Effects of calpain and Rho-kinase inhibitors on the acrosome reaction and motility of fowl spermatozoa in vitro

Reproduction ◽  
2006 ◽  
Vol 131 (1) ◽  
pp. 71-79 ◽  
Author(s):  
K Ashizawa ◽  
G J Wishart ◽  
S Katayama ◽  
D Takano ◽  
M Maeda ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Acrosome Reaction ◽  
Kinase Inhibitors ◽  
Rho Kinase ◽  
Specific Inhibitor ◽  
Immunoblot Analysis ◽  
Calpain Inhibitor ◽  
Cytoplasmic Matrix ◽  
Sperm Extract

At the avian body temperature of 40 °C, intact fowl spermatozoa require Ca2+for the initiation of motility and a combination of both Ca2+and homogenized inner perivitelline layer (IPVL) together to induce the acrosome reaction. Within the range of 1–100 μmol/l, neither PD 150606 (a Ca2+-dependent calpain inhibitor) nor Y-27632 (an inhibitor of Ca2+-dependent Rho-kinase) were able to inhibit the acrosome reaction induced by the presence of Ca2+and IPVL. However, PD 150606, although not Y-27632, was able to inhibit sperm motility initiated by Ca2+, as well as motility initiated by calyculin A – a specific inhibitor of protein phosphatases, which also initiates sperm motility at 40 °C. The addition of PD 150606 did not reduce the ATP concentrations of intact spermatozoa, nor the motility of demembranated spermatozoa. Immunoblot analysis of sperm extract using a polyclonal antibody against calpain 12 revealed a cross-reacting protein of approximately 80 kDa. These results suggest that Rho-kinase is not involved in the regulation of the acrosome reaction or of motility in fowl spermatozoa. In contrast, calpain appears to be involved in the regulation of flagellar movement, but not izn that of the acrosome reaction. Furthermore, it seems that endogenous calpain is present in the cytoplasmic matrix and/or the plasma membrane, but not retained in the axoneme and/or accessory cytoskeletal components.

scholarly journals Sperm cryopreservation of lane snapper Lutjanus synagris(Linnaeus, 1758)

2015 ◽  
Vol 75 (3) ◽  
pp. 662-669 ◽  
Author(s):  
EG Sanches ◽  
IR Oliveira ◽  
PCS Serralheiro ◽  
VR Cerqueira
Keyword(s):  
Sperm Motility ◽  
Sperm Quality ◽  
Equilibration Time ◽  
Dilution Ratio ◽  
Sperm Cryopreservation ◽  
Ph Values ◽  
Cryopreserved Sperm ◽  
Time Density ◽  
Motility Rate

AbstractThis study aims developing and evaluate a protocol of semen cryopreservation of the lane snapper Lutjanus synagris. Firstly, sperm motility rate, motility time, density and spermatocrit were appraised to characterize the sperm quality of the lane snapper. The effect of three extenders with distinct ionic compositions and pH values combined with seven concentrations of cryoprotector dimethylsulfoxide (0; 2.5; 5.0; 7.5; 10.0; 12.5 e 15.0%), five cooling rates (110, 90, 60, 45 e 30°C –min), nine equilibration time (1; 2,5; 5; 10; 15; 20; 25; 30 e 60 minutes) e five dilutions ratio (1:1; 1:3; 1:6; 1:10 e 1:20) on the sperm motility rate and motility time were analyzed. Fertilization test was accomplished to evaluate the viability of the cryopreserved sperm. The higher sperm motility rate and motility time (P<0.05) was achieved by combining extender with pH 8.2 with 10% concentration of dimethylsulfoxide and cooling rate 60°C –min, 1 minute of equilibration time and 1:3 (v/v) dilution ratio. The use of cryopreserved sperm presented fertilization rates >60% validating the present protocol for lane snapper. The cryoconserved sperm of lane snapper is a viable alternative, being possible to maintain appropriate sperm viability.

scholarly journals Effect of dietary supplementation with omega-3 and -6 on fresh and frozen/thawed sperm quality of dogs

2017 ◽  
Vol 38 (5) ◽  
pp. 3069 ◽  
Author(s):  
Ana Carolina Rodrigues ◽  
Camila Montanari Ruiz ◽  
Carla Daniela Dan De Nardo ◽  
Gabriele Barros Mothé ◽  
Fabiano Martinez Rossi ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Sperm Motility ◽  
Sperm Quality ◽  
Dietary Supplementation ◽  
Reproductive Efficiency ◽  
Omega 3 ◽  
Oral Supplementation ◽  
Animal Reproduction ◽  
Fresh Semen

For years, fatty acids have been recommended as a dietary supplement to improve canine hair. For animal reproduction, supplementation with omegas has been used to increase the reproductive efficiency and conception rate, but few studies have been conducted in dogs. The aim of this study was to evaluate the effects of daily dietary supplementation with omega-3 and -6 on the quality of fresh and frozen/thawed semen in canines. Semen was collected from seven dogs and evaluated for sperm motility, vigor, concentration, and morphology. The 17-week study included 119 ejaculates and was divided according to oral supplementation with omega-3 and -6: M1 (1st-5th week) or pre-supplementation; M2 (6th-9th week) and M3 (10th-13th week) or during supplementation; and M4 (14th-17th week) or post-supplementation. After analysis, the semen was frozen and then revaluated both immediately and 30 minutes (at 37° C) after thawing. Supplementation with omegas increased sperm motility, vigor, and concentration; however, supplementation had no influence on semen freezability. In addition, there was no improvement in sperm motility after supplementation when the thawed cells were maintained at 37° C for 30 minutes. We concluded that dietary supplementation with omega-3 and -6 for 4 to 8 weeks can improve the quality of fresh semen, although it has no effect on the freezability of canine semen.

Seminal plasma proteins protect flow-sorted ram spermatozoa from freeze - thaw damage

2009 ◽  
Vol 21 (4) ◽  
pp. 571 ◽  
Author(s):  
T. Leahy ◽  
J. I. Marti ◽  
G. Evans ◽  
W. M. C. Maxwell
Keyword(s):  
Seminal Plasma ◽  
High Speed ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Weight Fraction ◽  
Protein Supplementation ◽  
Freezing Process ◽  
Spermatozoa Motility ◽  
Functional Integrity

Seminal plasma improves the functional integrity of compromised ram spermatozoa but has been reported to be toxic to sorted spermatozoa. The present study attempted to clarify this paradoxical effect and improve the functional integrity of spermatozoa following sorting and cryopreservation. The in vitro function of sorted spermatozoa (motility characteristics and membrane integrity) was examined after supplementation with differing concentrations and protein fractions of seminal plasma at various stages of the sorting and freezing process. For all experiments, spermatozoa (two males, n = four ejaculates per male) were processed through a high-speed flow cytometer before cryopreservation, thawing and incubation for 6 h (37°C). Supplementation of crude seminal plasma (CP), its low molecular weight fraction (LP; <10 kDa) or protein-rich fraction (SPP; >10 kDa), immediately before freezing improved the functional integrity of sorted spermatozoa compared with no supplementation (control), whereas supplementation after thawing had no effect for CP and LP. The protective effect of seminal plasma was not altered by increasing the amount of protein supplementation. No toxic effect of CP, SPP or LP was evident even when supplemented at high protein concentrations. It is concluded that seminal plasma protein, if added to ram spermatozoa after sorting and before freezing, can improve post-thaw sperm quality and consequently the efficiency of sorting. This effect is most likely related to protection of the spermatozoa during freeze–thawing.

scholarly journals Spectrophotometric Analysis of the Resazurin Reduction Test as a Tool for Assessing Canine Semen Quality

2013 ◽  
Vol 57 (2) ◽  
pp. 281-285 ◽  
Author(s):  
Rafał Strzeżek ◽  
Krystyna Filipowicz ◽  
Marta Stańczak ◽  
Władysław Kordan
Keyword(s):  
Sperm Motility ◽  
Seminal Plasma ◽  
Semen Quality ◽  
Membrane Integrity ◽  
Spectrophotometric Analysis ◽  
Normal Morphology ◽  
Additional Tool ◽  
Reduction Test ◽  
Highly Correlated

Abstract The resazurin reduction test (RRT) was subjected to spectrophotometric analysis to evaluate the quality of canine semen. Twenty four samples of canine semen were analysed. The absorption peaks for resazurin and resorufin were determined at 615 and 580 nm, respectively. The RRT ratio (RRTsperm-the ratio for samples containing spermatozoa, RRTplasma-the ratio for samples containing seminal plasma) was calculated by dividing the absorbance at 580 nm by the absorbance at 615 nm. Spearman’s correlation test was used to determine the significance of correlations between the analysed sperm parameters and the results of the resazurin reduction assay. The RRT ratio was highly correlated with sperm motility (r=0.68, P<0.01), progressive sperm motility (r=0.61, P<0.01), the subpopulation of cells with rapid velocity (r=0.72, P<0.01), and the subpopulation of cells with medium velocity (r= -0.54, P<0.05). A negative correlation was observed between the reducing capacity of seminal plasma vs. sperm with plasma membrane integrity (r= -0.60, P<0.01) and sperm with normal morphology (r= -0.58, P<0.01). The RRT test can be used as an additional tool for evaluation of the quality of canine semen.

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