scholarly journals Comparative effects of addition of superoxide dismutase and reduced glutathione on cryopreservation of Sahiwal bull semen

2019 ◽  
Vol 69 (4) ◽  
pp. 1291
Author(s):  
A. Murtaza ◽  
M. Ahmad ◽  
M. Zubair ◽  
S. Umar ◽  
A. Mushtaq ◽  
...  
Keyword(s):  
Superoxide Dismutase ◽  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Semen Quality ◽  
Reduced Glutathione ◽  
Membrane Integrity ◽  
Bull Semen ◽  
Microscopic Evaluation ◽  
Artificial Vagina

The present study aimed to investigate effects of superoxide dismutase (SOD) and reduced glutathione (GSH) on the quality of frozen-thawed semen of Sahiwal bulls. Semen was collected twice a week for 8 weeks by artificial vagina from six Sahiwal bulls, kept at the Semen Production Unit Qadirabad, Sahiwal-Pakistan. After gross and microscopic evaluation, qualifying semen ejaculates were divided into 10 equal aliquots and diluted in extenders enriched with no antioxidants (control); or supplemented with either SOD (50, 100 and 200 IU/mL), or GSH (0.5, 1 and 2 mM) or their combinations (50 IU/mL SOD and 0.5 mM GSH, 100 IU/mL SOD and 1 mM GSH and 200 IU/mL SOD and 2 mM GSH). Samples were then frozen and stored in liquid nitrogen at -196°C for 24 h. The following parameters were evaluated for semen quality: post-thawed sperm motility, viability, acrosome and membrane integrity. According to the results, sperm motility, viability, acrosome and membrane integrity were significantly (P<0.05) higher in samples treated either with 100 IU/mL of SOD; 1 mM and 2 mM of GSH or 50 IU/mL of SOD plus 0.5 mM of GSH. In conclusion, semen quality might be improved by supplementing semen extenders with 100 IU/mL of SOD; 0.5 and 1 mM of GSH and combination of 50 IU/mL and 0.5 mM of SOD and GSH, respectively.

Effect of docosahexanoic acid on quality of frozen–thawed bull semen in BioXcell extender

2017 ◽  
Vol 29 (3) ◽  
pp. 490 ◽  
Author(s):  
Asmatullah Kaka ◽  
Wahid Haron ◽  
Rosnina Yusoff ◽  
Nurhusien Yimer ◽  
A. M. Khumran ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Optimum Level ◽  
Docosahexanoic Acid ◽  
Normal Morphology ◽  
Bull Semen ◽  
Acrosome Integrity

This study was conducted to investigate the effect of docosahexanoic acid (DHA) supplementation in BioXcell extender on the quality of frozen–thawed bull semen. Twenty-four ejaculates were collected from three bulls (eight from each bull). Ejaculates with motility ≥70% and normal morphology ≥80% were extended into BioXcell extender to which 0 (control), 3, 5, 10 or 15 ng mL–1 DHA was added. The supplemented semen samples were incubated at 37°C for 15 min for DHA uptake by spermatozoa. Later, samples were cooled for 2 h at 5°C and packaged into 0.25-mL straws, frozen in liquid nitrogen for 24 h and subsequently thawed for evaluation. Results are presented as percentages ± s.e.m. Supplementation with DHA at 3 ng mL–1 significantly improved sperm functional parameters including sperm motility, normal morphology, viability, acrosome integrity and membrane integrity when compared with other supplemented groups and the control. Lipid peroxidation increased as the incorporation of DHA supplementation increased. In conclusion, 3 ng mL–1 concentration of DHA resulted in superior quality of frozen–thawed bull spermatozoa and is suggested as the optimum level of DHA to be added into BioXcell extender.

Quality of ram spermatozoa separated with modified swim up method

2018 ◽  
Vol 33 (2) ◽  
pp. 62-70 ◽  
Author(s):  
A Hossain ◽  
MM Islam ◽  
F Naznin ◽  
RN Ferdousi ◽  
FY Bari ◽  
...  
Keyword(s):  
Semen Quality ◽  
Membrane Integrity ◽  
Quality Parameters ◽  
Fluid Medium ◽  
Normal Morphology ◽  
The Mean ◽  
Mass Activity ◽  
Fresh Semen ◽  
Artificial Vagina

Semen was collected from four rams, using artificial vagina and viability%, motility% and plasma membrane integrity% were measured. Fresh ejaculates (n = 32) were separated by modified swim-up separation using modified human tubal fluid medium. Four fractions of supernatant were collected at 15-minute intervals. The mean volume, mass activity, concentration, motility%, viability%, normal morphology and membrane integrity% (HOST +ve) of fresh semen were 1.0 ± 0.14, 4.1 ± 0.1 × 109 spermatozoa/ml, 85.0 ± 1.3, 89.4 ± 1.0, 85.5 ± 0.7, 84.7 ± 0.5 respectively. There was no significant (P>0.05) difference in fresh semen quality parameters between rams. The motility%, viability% and HOST +ve % of first, second, third and fourth fractions were 53.4 ± 0.5, 68.2 ± 0.3, 74.8 ± 0.3 and 65.5 ± 0.4; 55.5 ± 0.4, 66.2 ± 0.4, 74.5 ± 0.3 and 73.6 ± 0.3 and 66.7 ± 0.5, 66.8 ± 0.5, 65.2 ± 0.4 and 74.7 ± 0.5 respectively. The motility%, viability% and membrane integrity% of separated semen samples differed significantly (P<0.05) between four fractions. The mean motility% and viability% were significantly higher (P<0.05) in third fraction (74.8 ± 0.3%), whereas the mean HOST +ve% was significantly higher (P<0.05) in fourth fraction (74.7 ± 0.5). All quality parameters of separated spermatozoa were significantly (P<0.05) lower than that of fresh semen. The pregnancy rates were higher with fresh semen (71%) in comparison to that of separated sample (57%).Bangl. vet. 2016. Vol. 33, No. 2, 62-70

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.

scholarly journals Effect of Supplementation of Polyvinylpyrrolidone in Extender on Buffalo Semen Parameters

2020 ◽  
Vol 53 (1) ◽  
Author(s):  
Bushra Ismail Khan ◽  
Shamim Akhter ◽  
Sanwal Aslam ◽  
Rabea Ejaz
Keyword(s):  
Sperm Motility ◽  
Semen Quality ◽  
Membrane Integrity ◽  
Bubalus Bubalis ◽  
Semen Parameters ◽  
Suction Pump ◽  
Bull Semen ◽  
Buffalo Semen ◽  
Live Sperm ◽  
And Control

The current study was planned to evaluate the supplementation of Polyvinylpyrrolidone in extender on cryopreservation of Nili-Ravi buffalo bull semen. The semen samples were collected from Nili-Ravi buffalo (Bubalus bubalis) bull kept at SPU Qadirabad, District Sahiwal, Pakistan. Qualifying semen ejaculates having motility >60%, volume >5-6ml and concentration >0.5 billion/ ml were diluted 50 × 106 motile sperm ml approximately at 37°C in Tris-citric acid extender supplemented with different concentrations of PVP (0.01, 0.05, 0.1mM). The extender without PVP was kept as control. Semen was stored at 4°C for a period of 2 h and kept at 4°C for 4h. Semen was filled in 0.5 ml French straws using suction pump at 4°C, plunged and stored in liquid nitrogen (-196°C). Semen straws were rewarmed at 37°C for 30 seconds and assessed for sperm motility, plasma membrane integrity (PMI), dead sperm percentage and the live sperm percentage. The data on the role of PVP on different parameters of semen quality were analyzed by using ANOVA and RCBD. Higher percentage (P< 0.05) of sperm motility (66.1±7.51 and 59.4±10.72) and PMI (72.9±5.39 and 75.7±6.5) was observed in extenders having 0.05 mM and 0.1mM PVP compared to extenders having 1.5mM PVP and control. The percentage acrosomal integrity was observed greater (P< 0.05) in extended semen containing 0.1mM (68.2±0.50) PVP compared to extenders having 0.01 and control.

scholarly journals Effects of the addition of docosahexaenoic acid and ?-tocopherol on quality of equine spermatozoa stored at 5°C

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
Keyword(s):  
Lipid Peroxidation ◽  
Plasma Membrane ◽  
Docosahexaenoic Acid ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Control Group ◽  
Mitochondrial Activity ◽  
Membrane Composition ◽  
Artificial Vagina

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.

scholarly journals The quality of buck semen after feed additive minoxvit administration

2021 ◽  
Vol 19 (1) ◽  
pp. 87
Author(s):  
Anita Hafid ◽  
Riasari Gail Sianturi ◽  
Diana Andrianita Kusumaningrum ◽  
Yeni Widiawati ◽  
Anneke Anggraeni ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Semen Quality ◽  
Sperm Quality ◽  
Sperm Concentration ◽  
Positive Influence ◽  
Feed Additive ◽  
Concentration Data ◽  
Randomized Design ◽  
Microscopic Evaluation

<p class="MDPI17abstract"><strong>Objective: </strong>Reproduction and nutritional status is closely related. Nutritional deficiency or insufficiency directly affects reproductive efficiency. Deficiency of nutrition could affect the sperm quality and the ability to fertilize. The objective of this study was to evaluate the effect of feed additive (Minoxvit) on semen quality of buck.</p><p class="MDPI17abstract"><strong>Methods: </strong>This study used 6 sexually mature bucks, aged 1.5 years old. The bucks were fed daily with 3 kg of freshly chopped king grass, 1 kg of Legume (<em>Calliandra</em> sp.), and 500 g of concentrate. Three bucks were given Minoxvit by 1.25 g/day in the concentrate, while three other bucks were considered as control. Semen was evaluated once a week for 49 days. Semen was evaluated macroscopically and microscopically. The macroscopic evaluation consisted of volume, color, and consistency of semen, while microscopic evaluation consisted of mass motility, sperm motility, viability, and sperm concentration. Data were analyzed using Completely Randomized Design with Tukey test.<strong></strong></p><p class="MDPI17abstract"><strong>Results: </strong>The result showed significantly different (<em>P</em>&lt;0,05) in which bucks semen in Minoxvit addition group had higher semen volume (0.75 ml vs 0.54 ml), mass motility (3.32 vs 2.67), sperm motility (70% vs 58 %), sperm viability (86.67% vs 79.19%), and sperm consentration (2,6x10<sup>9</sup> mL vs 1,7x10<sup>9</sup> mL).<strong></strong></p><p><strong>Conclusions: </strong>This study concludes that the addition of Minoxvit has a positive influence on the quality of buck sperms providing volume, mass motility, individual motility, viability, and concentration of the sperm.</p>

Erratum to: 56 SINGLE LAYER CENTRIFUGATION THROUGH PURESPERM® 80 IMPROVES QUALITY OF CRYOPRESERVED DOG SPERMATOZOA

2013 ◽  
Vol 25 (1) ◽  
pp. 175
Author(s):  
L. Alcaráz ◽  
M. Hidalgo ◽  
M. J. Galvez ◽  
D. Acha ◽  
I. Ortiz ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Semen Quality ◽  
Semen Analysis ◽  
Gradient Centrifugation ◽  
Single Layer ◽  
Egg Yolk ◽  
Final Concentration ◽  
Viable Sperm

Density gradient centrifugation with PureSperm® (PureSperm® 40 + PureSperm® 80; Nidacon International, Mölndal, Sweden) has been satisfactorily used to enhance the quality of dog semen samples; however, no studies have been performed on the effect of single layer centrifugation (SLC) with PureSperm® on frozen–thawed dog semen. The aim of this study was to investigate if SLC with PureSperm® 80 can improve the post-thaw semen quality of dog. Semen from 5 dogs was collected by digital manipulation. Two ejaculates from each dog were centrifuged with Tris-based extender, supernatant was removed, and sperm pellet was suspended to a final concentration of 300–400 × 106 sperm mL–1 with CaniPROTM Freeze A plus 20% egg yolk at 22°C. Extended semen was cooled to 5°C within an hour and then diluted to a final concentration of 150–200 × 106 sperm mL–1 in CaniPROTM Freeze B plus 20% egg yolk at 5°C, loaded in 0.5-mL plastic straws and frozen horizontally in ranks placed 4 cm above the surface of liquid nitrogen vapors for 10 min, after which they were directly placed in liquid nitrogen. After 24 to 48 h of storage, straws were thawed in a water bath at 37°C for 30 s. After thawing, semen samples were divided in 2 aliquots: one of them was used as control and the other one was processed by SLC PureSperm® 80. Assessment of sperm motility (assessed by computerized-assisted semen analysis), morphology (Diff-Quick staining), and viability [triple fluorescent stain of propidium iodine/isothiocyanate-labeled peanut (Arachis hypogaea) agglutinin/Rhodamine 123] were evaluated in control and treated semen samples. Data were studied by ANOVA. Results are expressed as mean ± SEM. Significant (P < 0.001) differences were found between SLC-treated and control semen for sperm motility (percentage of total motile spermatozoa: 93.65 ± 0.05 v. 83.79 ± 0.13; percentage of progressive motile spermatozoa: 79.38 ± 6.66 v. 54.61 ± 16.11), morphology (86.45 ± 0.01 v. 83.51 ± 0.01), and viability (percentage of viable sperm with an intact acrosome: 58.32 ± 0.04 v. 36.50 ± 0.17; percentage of viable sperm with an acrosome reaction: 2.81 ± 0.01 v. 9.74 ± 0.21). Based on our results, we can conclude that SLC with PureSperm® 80 is an alternative and successful method for improving the quality of frozen–thawed dog spermatozoa, selecting good-quality spermatozoa (motile, morphologically normal, viable, and acrosome intact spermatozoa) from the rest of the semen sample.

Evaluate the fresh and Post Thawing Quality of Holstein Friesian Bull semen

2019 ◽  
Vol 1 (1) ◽  
pp. 22-26
Author(s):  
Jamal Mehmood shah ◽  
Tahir Hameed ◽  
Farhat abbas Bokhari ◽  
Gul zaman
Keyword(s):  
Sperm Motility ◽  
Sperm Concentration ◽  
Membrane Integrity ◽  
Pooled Data ◽  
Bull Semen ◽  
Holstein Friesian ◽  
Semen Volume ◽  
Sperm Membrane ◽  
Mass Activity

TThe   study   was  carried   out   during the yearr  2017   to   determine   the   fresh   and   post-thaw   quality   of   four   Holstein   Friesian   bulls.   Semen   samples   were   examined   for ejaculate   volume,   pH,   mass   activity,   sperm   concentration   and   sperm   motility and   sperm   membrane   integrity   (HOST).   The   ejaculated   semen   volume   of   Holstein   Friesian   bulls   was   in   the   range   of   6-8ml   and   differences   in   ejaculates   were   significant   (P<0.01).   The   volume   was   significantly   (P<0.05)   higher   (7.75ml)   when   collected   on   5th   July,   slightly   decrease   in   volume   (7.25ml)   when   collected   on   12th   May   and   26th   July.   Semen   pH   was   higher   (6.65)   for   26th   July   ejaculation   and   lowest (6.55ml)   for   24th   May   ejaculation.   The   results   indicated   that   the   semen   of   Holstein   Friesian   bulls   did   not   have   considerable   variation   in   pH   during   May   –   July.   Most   of   the   semen   samples   were   creamy   white and   yellow   in   colour,   while   few   samples   were   milky   white   and   watery.   Mass   activity   score   of   semen   samples   indicated   vigorous   movement with   moderate rapid   waves   and   eddies.   The   sperm   motility   of   fresh   vs   post-thaw   semen   was   obtained   72.80   vs   48.75   percent   (12th   May),   75.00   vs   48.75   percent   (24th   May),   76.25   vs   55.00   percent   (2nd   June),   73.75   vs   52.50   percent   (21st   June),   75.00   vs   56.25   (5th   July)   and   75.00   vs   50.00   percent   (26th   July).   In   case   of   post-thaw   semen,   highest   sperm   motility   of   56.25   percent   was   recorded   in   semen   ejaculated   on   5th   July,   while   lowest   post-thaw   sperm   motility   of   48.75   percent   was   observed   in   semen   ejaculated   on   12th   May   and   24th   May.   Highest   sperm   concentration   of   1549.75x106   was   determined   in   semen   ejaculated   on   21.06.2017   and   lowest   concentration   (1259.50x106)   in   26.07.2017   collected   samples.   It   was   concluded   that   sperm   motility   was   significantly   (P<0.01)   affected   by   semen   types   (fresh   and   post-thaw),   while   sperm   concentration   was   also   significantly   (P<0.01)   affected   by   ejaculation   date   and   bulls.   The   membrane   integrity   (HOST)   of   the   pooled   data   on   fresh   semen   samples   over   a   period   of   three   months   was   58.37   percent   against   post-thaw   membrane   integrity   of   44.79   percent.

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 Erratum to: 56 SINGLE LAYER CENTRIFUGATION THROUGH PURESPERM® 80 IMPROVES QUALITY OF CRYOPRESERVED DOG SPERMATOZOA

2013 ◽  
Vol 25 (3) ◽  
pp. 587
Author(s):  
L. Alcaráz ◽  
M. Hidalgo ◽  
M. J. Galvez ◽  
D. Acha ◽  
I. Ortiz ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Semen Quality ◽  
Semen Analysis ◽  
Gradient Centrifugation ◽  
Single Layer ◽  
Egg Yolk ◽  
Final Concentration ◽  
Viable Sperm

Density gradient centrifugation with PureSperm® (PureSperm® 40+PureSperm® 80; Nidacon International, Mölndal, Sweden) has been satisfactorily used to enhance the quality of dog semen samples; however, no studies have been performed on the effect of single layer centrifugation (SLC) with PureSperm® on frozen–thawed dog semen. The aim of this study was to investigate if SLC with PureSperm® 80 can improve the post-thaw semen quality of dog. Semen from 5 dogs was collected by digital manipulation. Two ejaculates from each dog were centrifuged with Tris-based extender, supernatant was removed, and sperm pellet was suspended to a final concentration of 300–400×106spermmL–1 with CaniPROTM Freeze A plus 20% egg yolk at 22°C. Extended semen was cooled to 5°C within an hour and then diluted to a final concentration of 150–200×106spermmL–1 in CaniPROTM Freeze B plus 20% egg yolk at 5°C, loaded in 0.5-mL plastic straws and frozen horizontally in ranks placed 4cm above the surface of liquid nitrogen vapors for 10min, after which they were directly placed in liquid nitrogen. After 24 to 48h of storage, straws were thawed in a water bath at 37°C for 30s. After thawing, semen samples were divided in 2 aliquots: one of them was used as control and the other one was processed by SLC PureSperm® 80. Assessment of sperm motility (assessed by computerized-assisted semen analysis), morphology (Diff-Quick staining), and viability [triple fluorescent stain of propidium iodine/isothiocyanate-labeled peanut (Arachis hypogaea) agglutinin/Rhodamine 123] were evaluated in control and treated semen samples. Data were studied by ANOVA. Results are expressed as mean ± SEM. Significant (P&lt;0.001) differences were found between SLC-treated and control semen for sperm motility (percentage of total motile spermatozoa: 93.65±0.05 v. 83.79±0.13; percentage of progressive motile spermatozoa: 79.38±6.66 v. 54.61±16.11), morphology (86.45±0.01 v. 83.51±0.01), and viability (percentage of viable sperm with an intact acrosome: 58.32±0.04 v. 36.50±0.17; percentage of viable sperm with an acrosome reaction: 2.81±0.01 v. 9.74±0.21). Based on our results, we can conclude that SLC with PureSperm® 80 is an alternative and successful method for improving the quality of frozen–thawed dog spermatozoa, selecting good-quality spermatozoa (motile, morphologically normal, viable, and acrosome intact spermatozoa) from the rest of the semen sample.

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