138 EFFECT OF SEMINAL PLASMA ON CHILLING AND FREEZING OF CANINE SPERMATOZOA

2007 ◽  
Vol 19 (1) ◽  
pp. 187
Author(s):  
I. Yu ◽  
Y. J. Kim ◽  
I. S. Kim ◽  
S. P. Leibo ◽  
N. Songsasen
Keyword(s):  
Pisum Sativum ◽  
Seminal Plasma ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Beneficial Effects ◽  
Progressive Motility ◽  
Healthy Dogs ◽  
Sperm Survival ◽  
Acrosome Integrity ◽  
Group 1

Seminal plasma (SP) is usually removed from semen that is to be cryopreserved. However, some reports indicate that SP has beneficial effects on spermatozoa during chilling and freezing (Barrios et al. 2000 Biol. Reprod. 63, 1531–1537; Moore et al. 2005 Theriogenology 63, 2372–2381; Vadnais et al. 2005 Anim. Reprod. Sci. 87, 121–132). The purpose of this study was to determine the effect on sperm survival of adding SP to the extender before cooling and freezing canine spermatozoa. In replicate experiments, ejaculates obtained from 4 healthy dogs (3–4 years old) of various breeds were pooled and centrifuged at 300g for 10 min at 25�C; the supernatant of seminal plasma was decanted. Spermatozoa were suspended in egg yolk-Tris (EYT) buffer. The study comprised 2 experiments: Exp. 1: Sperm were suspended in EYT extender containing 0%, 20%, 50%, 80%, or 100% SP, and were slowly cooled to 4�C for 2 h or held at 25�C as controls. Exp. 2: Sperm samples, each of which contained 1 � 108 sperm mL-1, were assigned to 5 groups to be frozen. In group 1, sperm in EYT + 20% SP were cooled to 4�C, diluted to contain final concentrations of 5% glycerol + 10% SP in EYT, and then frozen. In the 4 other groups, sperm in EYT alone were first cooled slowly to 4�C, then diluted to contain 5% glycerol plus 0%, 20%, 40%, or 50% SP in EYT, and then frozen. Spermatozoa were frozen at 25�C min in plastic straws that were suspended above liquid nitrogen and thawed in water at 38�C for 30 s. Sperm survival was assayed by determining progressive motility and integrity of plasma and acrosome membranes. Progressive motility was determined by microscopic examination at 400� magnification. Membrane integrity was assessed by use of a double fluorescent dye, and acrosome integrity by staining sperm with Pisum sativum agglutinin. The results of the first experiment showed that 20%, 50%, 80%, or 100% SP did not improve motility, membrane integrity, or acrosome integrity of spermatozoa chilled to 4�C compared to those chilled without SP (P > 0.05). Survival of spermatozoa suspended in EYT + 20% SP and maintained at 25�C was significantly higher than for those that were chilled (P < 0.05). The results of the second experiment showed that spermatozoa suspended in EYT + 20% SP and then diluted at 4�C to contain 5% glycerol + 10% SP exhibited the highest progressive motility and membrane integrity after being frozen and thawed (P < 0.05). In summary, although seminal plasma did not affect spermatozoa that were only chilled, addition of seminal plasma did significantly improve survival of canine spermatozoa that were frozen and thawed.

90 EFFECT OF EGG YOLK ON THE KINEMATICS AND ACROSOME MEMBRANE INTEGRITY OF COOLED-REWARMED CANINE SPERMATOZOA

2010 ◽  
Vol 22 (1) ◽  
pp. 204
Author(s):  
J. Dorado ◽  
M. J. Galvez ◽  
M. R. Murabito ◽  
S. Demyda ◽  
L. J. De Luca ◽  
...  
Keyword(s):  
Semen Analysis ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Computer Assisted ◽  
Mean Values ◽  
Head Displacement ◽  
Healthy Dogs ◽  
Acrosome Integrity ◽  
Digital Manipulation ◽  
Lateral Head

Tris-egg yolk-based diluents provide adequate cryoprotection for the sperm of most species. This study was conducted to compare the ability of Tris-glucose extender containing 2 different concentrations of egg yolk to maintain sperm motility and acrosome integrity of canine spermatozoa during 72 h of preservation. For this purpose, a total of 20 ejaculates from 4 clinically healthy dogs (2 Spanish Greyhound, 1 German Pointer, and 1 Crossbreed) were collected by digital manipulation. The sperm-rich fraction of each ejaculate was divided into 2 aliquots. Then, they were diluted in Tris-based extender and centrifuged at 700g for 8 min. Sperm pellets were resuspended in either Tris buffer added to 20% (EY20) or 10% centrifuged egg yolk (EY10) and cooled to 5°C over 72 h. The effects of these extenders on motility and acrosome integrity were assessed objectively using a computer-aided semen analyzer (Sperm Class Analyzer, Microptic SL, Spain) and Spermac® staining, respectively. Each cooled-rewarmed semen sample was evaluated after 24, 48, and 72 h of preservation. Sperm motion parameters shown by computer-assisted semen analysis (CASA) are progressively motile (PMS) and motile spermatozoa (MS), curvilinear velocity (CLV), average path velocity (APV), progressive speed (SLV), and lateral head displacement (LHD). Data were statistically analysed by ANOVA. Dependent variables expressed as percentages were arsine-transformed before analysis. Differences between mean values were evaluated by the Duncan method. Data were presented as mean ± SEM. Differences were considered significant when P < 0.05. Analyses were performed using the statistical package SPSS 12.0. A total of 98 172 motile sperm trajectories were analyzed by CASA: 52 259 in EY20 and 45 913 in EY10. After 24, 48, and 72 h of preservation, MS and PMS were statistically higher (P < 0.01) in EY20. No significant differences were found for LHD using either extender over a 72-h period. No significant differences were observed for CLV using either extender during the first 2 days. At Day 3, CLV data were significantly higher (P < 0.01) in EY20. Similarly, from Day 2, APV was significantly higher (P < 0.001) in EY20. After 24 h of preservation, SLV was statistically higher (P < 0.001) in EY10, whereas the opposite tendency was found at Day 3. No significant differences were observed for SLV using either extender after 48 h of preservation. During the first 2 days, acrosome integrity was statistically higher (P < 0.001) in EY20. At hour 72, higher acrosome integrity (P < 0.001) was observed in EY10. In conclusion, we have observed that the EY20 extender provided higher motility after 72 h of chilled preservation; however, the acrosome membrane integrity was better preserved in EY10.

113 EFFECT OF SPERM COATING ON THE QUALITY OF BOVINE FROZEN - THAWED SPERMATOZOA

2006 ◽  
Vol 18 (2) ◽  
pp. 164
Author(s):  
M. Thys ◽  
A. Van Soom ◽  
J. Dewulf ◽  
T. Rijsselaere ◽  
A. de Kruif
Keyword(s):  
Seminal Plasma ◽  
Sperm Quality ◽  
Univariate Analysis ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Freezing And Thawing ◽  
Bovine Sperm ◽  
Sperm Characteristics ◽  
Group 2 ◽  
Group 1

The substantial decrease of sperm quality after cryopreservation remains an important issue in the artificial insemination industry. Sperm coating with Triladyl® (Minitübe, Tiefenbach, Germany) during ejaculation can preserve sperm characteristics and oocyte penetrating capacity of fresh bovine spermatozoa stored in egg yolk diluent for up to 6 days (De Pauw et al. 2003 Theriogenology 59, 1109–1122). Since collecting semen in a tube containing egg yolk-Tris extender (sperm coating) limits the period of contact between spermatozoa and seminal plasma, the present experiment was conducted to assess if this slightly adjusted method of sperm collection could also have a significant effect on bovine sperm quality after cryopreservation. Semen of five young Holstein Friesian bulls was collected by means of an artificial vagina connected to an empty tube (Group 1; five ejaculates per bull) or a tube containing 4 mL of an egg yolk-Tris extender (Groups 2 and 3; each five ejaculates per bull). The semen samples of Group 1 were conventionally diluted in straws (60 × 106 sperm/mL), frozen, and stored in liquid nitrogen. The samples of Group 3 were centrifuged, and after removing diluent and seminal plasma, the sperm pellet was conventionally diluted and processed. The samples of Group 2 were processed without removal of the supernatant. After thawing each ejaculate was analyzed for average path velocity (VAP), beat cross frequency (BCF), and progressive motility (PROG) using CASA (Minitübe, Tiefenbach, Germany). Furthermore, the membrane integrity of each sample was evaluated using fluorescent SYBR®–14/PI staining (BD Biosciences, Erembodegem, Belgium). All parameters were compared among the three groups of sperm using univariate analysis of variance (SPSS 12.0; SPSS, Inc., Chicago, IL, USA). No significant differences could be observed among the three groups for all of the evaluated sperm characteristics (Table 1). A significant effect of the bull could be determined for all analyzed parameters (P ≤ 0.02), except for the percentage of moribund cells. Nevertheless, the group-bull interaction was never statistically significant. Coating bovine sperm with an egg yolk-Tris extender during ejaculation cannot prevent the substantial deterioration of the spermatozoa that occurs during freezing and thawing since this method of sperm collection does not significantly influence the motility parameters or the membrane integrity after thawing. Table 1. VAP, BCF, PROG, and percentage of membrane-intact, dead, and moribund spermatozoa for the three groups of sperm This research was supported by IWT (no. IWT/020727).

scholarly journals 92THE ADDITION OF SEMINAL PLASMA FROM INDIVIDUAL BOARS TO FREEZING EXTENDER CAN IMPROVE THE POST-THAW SPERM SURVIVAL

2004 ◽  
Vol 16 (2) ◽  
pp. 167 ◽  
Author(s):  
T. Cremades ◽  
G. Carvajal ◽  
M. Hernandez ◽  
J.M. Vazquez ◽  
E.A. Martinez ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Potential Effect ◽  
Conclusive Evidence ◽  
Sperm Cryopreservation ◽  
Low Concentrations ◽  
Sperm Survival ◽  
And Control ◽  
Boar Spermatozoa

Contradictory results have been reported about the effect of seminal plasma (SP) on the freezability of mammalian spermatozoa. In pigs, current methods for sperm cryopreservation involve removing seminal plasma. Therefore, no conclusive evidence of the potential effect of SP on the freezability of boar spermatozoa has been reported. In this study, we evaluate the effect of the addition of low concentrations of SP from individual boars to the freezing extender on post-thaw sperm survival. Sperm cryopreservation procedure included: dilution of sperm-rich fraction in Beltsville Thaw Solution extender (BTS), cooling to 17°C for 16h, centrifugation at 2400g for 3min, dilution in lactose/egg-yolk/glycerol/Equex Stem (freezing extender) to a final concentration of 1×109 spermmL−1, dispensing into 0.5-mL straws, and freezing in a programmable cell freezer at 20°Cmin−1. Thawing was carried out in a waterbath at 37°C for 20s. Post-thaw sperm survival was assessed by progressive sperm motility (PSM) using a CASA system (SCA); plasma membrane integrity (PMI) and acrosome membrane integrity (AMI) were assessed by flow cytometric procedures (SYBR-14/PI and FITC-PNA/PI, respectively) at 30 and 150min post-thawing in BTS-diluted thaw spermatozoa held in a waterbath at 37°C. Four individual seminal plasma donors (SP1 to SP4) were selected in a preliminary study in which 48 ejaculates from 12 boars (4 ejaculates/boar) were cryopreserved. Then the boars were classified into 3 groups (good, moderate and bad freezers) based on their post-thaw sperm survival. SP1 and SP2 were good freezers (&gt;60% PSM and PMI), SP3 was a moderate freezer (40–60% PSM and PMI) and SP4 was a bad freezer (&lt;40% PSM and PMI). In the main experiment, pooled sperm-rich fractions collected from 9 mature hybrid boars were divided into five aliquots and each was diluted with freezing extender supplemented with 0% (control) or 10% of SP (1–4). Data from eight replicates were analyzed as a split plot design using a PROMIXED model. The addition of SP to freezing extender had a significant effect (P&lt;0.05) on post-thaw sperm survival compared to control. Moreover, there were significant differences (P&lt;0.05) between SP donors. PSM, PMI and AMI were significantly (P&lt;0.05) higher in SP1 (56.71±4.30; 57.16±4.01 and 57.22±4.01, respectively) and SP2 (59.48±4.30; 60.17±4.01 and 60.05±4.01, respectively) compared to control (50.39±4.30; 49.98±4.01 and 49.54±4.01, respectively). There were no differences (P&gt;0.05) between SP3, SP4 and control. These results indicate that the addition of SP from particular boars (good freezers) to freezing extender may improve post-thaw sperm survival. Individual differences in the SP composition should explain the above results. Supported by INIA (RZ01-019) and MCYT (AGL2001-0471).

scholarly journals Effect of egg yolk-based extender and seminal plasma removal on sperm viability of cooled donkey semen

2021 ◽  
Vol 58 ◽  
pp. e174301
Author(s):  
Carolina Natalia Alonso ◽  
Catalina Castañeira ◽  
Ana Flores Bragulat ◽  
Luis Losinno
Keyword(s):  
Kinetic Parameters ◽  
Seminal Plasma ◽  
Membrane Integrity ◽  
Milk Proteins ◽  
Egg Yolk ◽  
Reproductive Efficiency ◽  
Sperm Viability ◽  
Progressive Motility ◽  
Live Sperm

Developing effective cooled semen protocols is essential to increase pregnancy rates and reproductive efficiency in donkeys. This study aimed to evaluate the effect on sperm kinetic parameters and membrane integrity in cooled donkey semen diluted with defined milk proteins extender with 1% or 2% of egg yolk and the removal of seminal plasma. Twenty-four ejaculates from six jackasses were collected. Each ejaculate was divided into four aliquots that were diluted in extender with 1% (EY1) or 2% (EY2) egg yolk. One sample from each group was centrifuged, seminal plasma was removed (CEY1, CEY2 groups, respectively), and the samples were then refrigerated at 5 °C for 24 h. Fresh and cooled semen samples were assessed for sperm motility, morphology, and plasma membrane integrity. Total motility, progressive motility, sperm kinetic parameters, or live sperm cells were not statistically different when semen was cooled with an extender supplemented with 1% or 2% of egg yolk. Seminal plasma removal does not affect total motility or sperm kinetic parameters. However, progressive motility decreased (P<0.05) when semen was extended with 2% of egg yolk and seminal plasma was removed. Membrane integrity was affected (P<0.05) in centrifuged samples. In conclusion, the obtained results suggest that there is no difference in sperm kinetics and membrane integrity when 1% or 2% of egg yolk was added to the Equiplus(R) extender. Also, the removal of seminal plasma by centrifugation did not have any beneficial effect on cooled donkey semen. Further studies are needed to relate these results with in vivo fertility tests with cooled donkey semen.

Supplementation of different concentrations of Orvus Es Paste (OEP) to ostrich egg yolk lipoprotein extender improves post-thaw boar semen quality

2014 ◽  
Vol 17 (2) ◽  
pp. 225-230 ◽  
Author(s):  
L. Fraser ◽  
E. Jasiewicz ◽  
W. Kordan
Keyword(s):  
Semen Quality ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Plasma Membrane Integrity ◽  
Sperm Characteristics ◽  
Boar Semen ◽  
Sperm Survival ◽  
Acrosome Integrity ◽  
Egg Yolk Lipoproteins

Abstract This study aimed to compare post-thaw quality of boar semen following freezing in an ostrich egg yolk lipoprotein (LPFo) extender supplemented with 0%, 0.25% and 0.50% Orvus Es Paste (OEP). Sperm assessments included total motility (TMOT), mitochondrial function (MF), plasma membrane integrity (PMI) and acrosome integrity (normal apical ridge, NAR). Considerable variations among boars and OEP treatments had a significant effect (P < 0.001) on post-thaw sperm characteristics. It was observed that post-thaw sperm characteristics were significantly compromised in semen samples frozen in the absence of OEP. By contrast, lactose-LPFo-glycerol extender supplemented with either 0.25% OEP or 0.50% OEP markedly enhanced post-thaw sperm characteristics. In all boars, there were no marked differences in post-thaw sperm TMOT between the freezing extenders supplemented with 0.25% and 0.50% OEP. However, a decline in the percentage of post-thaw motile spermatozoa was more pronounced in the extender supplemented with 0.50% OEP following a 120-min incubation period. Furthermore, the proportions of frozen-thawed spermatozoa with MF, PMI and NAR acrosomes varied significantly among the boars in the OEP-supplemented extenders. The findings of this study indicate that different OEP concentrations, in the presence of ostrich egg yolk lipoproteins, could have varying effects on post-thaw sperm survival

Effects of fulvic acids on goat sperm

Zygote ◽  
2018 ◽  
Vol 26 (3) ◽  
pp. 220-223
Author(s):  
Yu Xiao ◽  
Zhengmu Wu ◽  
Min Wang
Keyword(s):  
Superoxide Dismutase ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Fulvic Acids ◽  
Quality Parameters ◽  
Control Group ◽  
Progressive Motility ◽  
Acrosome Integrity

SummaryThe effects of adding fulvic acids (FAs) to semen extenders on the quality parameters of frozen–thawed goat buck spermatozoa remain undetermined. Buck semen samples collected from six mature goat bucks once a week were diluted with Tris–egg yolk-based extenders. The diluted semen samples were supplemented with FAs (0.2, 0.4 and 0.6%, w/w), cryopreserved, and evaluated for sperm-quality parameters. Addition of FAs to the extender increased progressive motility, acrosome integrity, membrane integrity, and superoxide dismutase and catalase activities and decreased percentage abnormality and sperm malondialdehyde level compared with the control group. However, excessive FA addition (>0.4%, w/w) to semen extenders did not improve the efficiency. The results indicated that FAs could be a promising cryoprotectant for goat buck sperm.

267 EFFECT OF SEMINAL PLASMA IN LAMA GLAMA SPERM

2015 ◽  
Vol 27 (1) ◽  
pp. 223 ◽  
Author(s):  
M. Carretero ◽  
F. Fumuso ◽  
M. Miragaya ◽  
C. Herrera ◽  
S. Giuliano
Keyword(s):  
Sperm Motility ◽  
Seminal Plasma ◽  
Membrane Integrity ◽  
South American ◽  
South American Camelids ◽  
Progressive Motility ◽  
Coomassie Blue ◽  
Blue Stain ◽  
Lama Glama ◽  
Motility Of Sperm

In South American camelids, raw semen only presents sperm with oscillatory movements. Therefore, it is necessary to treat these cells to enable them to acquire progressive motility. The effects of raw seminal plasma (SP) on sperm movement patterns (oscillatory, progressive, and hyperactive) have apparently not yet been reported. The objective of this study was to determine effects of raw seminal plasma on sperm motility, viability, and acrosomal status in fresh llama semen. A total of 15 ejaculates were collected (electroejaculation) from 5 llamas (n = 5, r = 3). Each ejaculate was diluted 4 : 1 in 0.1% collagenase in HEPES-TALP (HT) medium and incubated 4 min at 37°C, with the objective of separating spermatozoa from SP. Immediately after incubation, each ejaculate was divided into 2 and centrifuged for 8 min at 800 × g. Pellets were resuspended in either HT or raw SP and maintained at 37°C until evaluation (at 0, 1.5, and 3 h). Sperm motility was evaluated using a phase contrast microscope and a warm stage. Propidium iodide and carboxyfluorescein diacetate were used for assessing membrane integrity (viability). Acrosomal status was evaluated with the Coomassie blue stain. A split-plot design was used with treatment as a factor, with 2 levels (HT and SP) and time as the other factor, with 3 levels (0, 1.5, and 3 h), and blocked by males. There was no significant interaction between treatments (HT and SP) and times (0, 1.5, and 3 h) for each of the seminal characteristics evaluated. Progressive sperm motility was observed after collagenase treatment in all samples. Progressive motility disappeared immediately after the addition of raw SP and showed only oscillatory movements. In contrast, samples incubated in HT maintained progressive motility and became hyperactive. There were no differences (P > 0.05) in total motility of sperm incubated in HT among incubation times (0 h: 30.8 ± 18.9%; 1.5 h: 26.5 ± 11.5%; and 3 h: 21.5 ± 13.5%). However, in samples incubated with SP, a decrease (P < 0.05) in total sperm motility was detected after 3 h of incubation (0 h: 16.5 ± 12.6%, 3 h: 2.3 ± 3.2%). Sperm viability was not different (P > 0.05) between treatments (HT and SP); samples incubated in HT retained 78.4% of the initial viability (32.8/41.8, 3 h/0 h), and samples incubated in SP retained 69.7% of their initial viability (24.4/35.0, 3 h/0 h). The percentage of spermatozoa with intact acrosomes was not different (P > 0.05) between treatments (HT and SP); however, the percentage of sperm with intact acrosomes decreased after 3 h of incubation in both samples (HT and SP). Due to the presence of a high percentage of progressive and hyperactive motile sperm in samples incubated in HT and their absence in samples incubated in SP, we concluded that raw seminal plasma preserved oscillatory sperm motility. Further studies are needed to understand the effects of SP on South American camelid spermatozoa.

54 CRYOPRESERVATION OF DOMESTIC CAT EPIDIDYMAL SPERM IN A DEFINED EXTENDER WITHOUT ANIMAL OR PLANT PROTEINS

2012 ◽  
Vol 24 (1) ◽  
pp. 139
Author(s):  
J. R. Saenz ◽  
C. Dumas ◽  
B. L. Dresser ◽  
M. C. Gómez ◽  
R. A. Godke ◽  
...  
Keyword(s):  
Membrane Integrity ◽  
Egg Yolk ◽  
Domestic Cat ◽  
The Other ◽  
Initial Assessment ◽  
Computer Assisted ◽  
Plant Proteins ◽  
Epididymal Sperm ◽  
Sperm Suspension ◽  
Sperm Survival

Previously, we have shown that survival of cat sperm is maintained in both non-egg yolk, semi-defined extenders and in extenders with greatly reduced levels of egg yolk (2%). Usually, cryoprotectant is added to extended samples after gradual cooling to 4°C, but recent reports have shown that satisfactory sperm survival can be obtained after addition at 22°C. Here, our objectives were to examine sperm survival after (1) cryopreservation from 22°C vs after gradually cooling to 4°C or (2) cryopreservation in a completely defined extender without animal or plant proteins vs extender + 2% egg yolk. Epididymides from local veterinary clinics were dissected in HEPES 199 medium (He199). The sperm suspension was filtered (40 μ), layered onto a density gradient column and centrifuged at 650 × g for 20 min. Then, the sperm pellet was resuspended in 1 mL of He199 and centrifuged for 5 min at 800 × g and the subsequent pellet was extended in TEST Buffer with either 0% (0% EY) or 2% egg yolk (2% EY). Next, 0% EY samples were further split into 2 groups—either gradually cooled to 4°C before 12% glycerol (1:1) was added (4C-0%EY) or 12% glycerol (1:1) was added at 22°C without cooling (22C-0%EY). Control samples extended in 2% EY were cooled to 4°C before addition of 12% glycerol (1:1) (4C-2%EY). Samples were loaded into 0.25-mL straws and placed in a –80°C freezer for 20 min before storage in LN2. Sperm samples were thawed in air (22°C) for 5 s and immersed in a 60°C water bath for 5 s. After a 7-step addition of He199, samples were centrifuged at 800 × g for 5 min and pellets resuspended in He199. Sperm samples were evaluated for motility (Mot; computer-assisted semen analysis, 37°C) at 0 h (initial assessment), after cooling to 4°C (PC) and at 0-h (0-PT) and 3-h post-thaw (3-PT) incubation at 37°C. Membrane integrity (MI; SYBR 14-PI) and acrosomal status (AS; FITC-PNA) were analysed at the initial assessment, 0-PT and 3-PT. Results are shown in Table 1. At 4°C (PC), sperm extended in 0% EY and 2% EY maintained 92 and 91%, respectively, of their initial motility (66%). At 0-PT and 3-PT, motility in the 3 groups had decreased by >50% and >70%, respectively. Motility at 3-PT in the 22C-0%EY treatment was less than the other 2 treatments (P < 0.05; 1-way ANOVA). At 0-PT, samples in the 4C-2%EY group had a higher membrane integrity value (P < 0.05) than did the 22C-0%EY group, whereas that of the 4C-0%EY group was not different from the other 2 groups. However, at 3-PT, both groups cooled to 4°C before cryopreservation had higher membrane integrity values (P < 0.05) than the group cryopreserved at 22°C. At 0-PT and 3-PT, the percentage of sperm with intact acrosomes ranged from 69% (4C-2%EY) to 59% (22C-0%EY) and from 55% (4C-2%EY) to 43% (22C-0%EY) of the initial value (89%), respectively. In summary, we demonstrated that cat epididymal sperm could be frozen successfully in a completely defined TEST-buffered extender. Furthermore, we confirmed that addition of cryoprotectant (i.e. glycerol) after gradual cooling to 4°C is beneficial to post-thaw survival. Table 1.Motility (Mot), membrane integrity (MI) and acrosomal status (AS) of cat epididymal sperm before and after cryostorage

scholarly journals Comparison of Extenders With the Addition of Egg Yolk for Cooling Alpaca Sperm Obtained From Deferent Ducts

2020 ◽  
Vol 7 ◽  
Author(s):  
Mariana Lucía Bertuzzi ◽  
Edita Yola Torres ◽  
Teodosio Huanca ◽  
Deborah Neild ◽  
María Ignacia Carretero
Keyword(s):  
Sperm Motility ◽  
Chromatin Condensation ◽  
Egg Yolk ◽  
Final Concentration ◽  
Sperm Parameters ◽  
Membrane Function ◽  
Progressive Motility ◽  
Evaluation Time ◽  
Viable Sperm ◽  
Acrosome Integrity

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.

scholarly journals Effect of sunflower lecithin on Kalahari Red goat semen during cryopreservation

2018 ◽  
Vol 51 (1) ◽  
pp. 21-28 ◽  
Author(s):  
Sakirat Opeyemi Adeyanju ◽  
James Olatinbo Daramola ◽  
Jimoh Alao Olanite ◽  
Olufiropo Samson Awokola
Keyword(s):  
Soybean Lecithin ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Domestic Animal ◽  
Arginase Activity ◽  
Semen Parameters ◽  
Phospholipid Content ◽  
Acrosome Integrity ◽  
Semen Extender ◽  
Different Levels

Abstract Soybean lecithin had been used as an alternative to egg yolk in domestic animal semen extender during cryopreservation due to its characteristic phospholipid content which played a major cryoprotective role. This composition of soybean lecithin informed the replacement of soybean with sunflower lecithin (SL) in the extender for the Kalahari Red (KR) buck semen cryopreservation in this study. Effect of different levels of SL on the quality of the KR buck semen during cryopreservation using slow freezing method was evaluated. Semen samples were collected from four KR bucks of between two and two and half of age using artificial vagina, evaluated for motility and then diluted in extenders containing different levels of SL (1.5%, 3.0% and 4.5%) as experimental group and 0% SL or 20% egg yolk as control. Semen parameters including motility, acrosome integrity (AcI), membrane integrity (MI), malondialdehyde (MDA) concentration, cholesterol level and seminal arginase activity were evaluated for. The results showed that motility, acrosome integrity (AI) and membrane integrity were comparable at 0%, (22.00 ± 4.58, 82.00 ± 3.51 and 96.00 ± 2.03); 1.5%, (23.00 ± 2.08, 87.00 ± 3.79 and 89.00 ± 2.08); 3.0%, (13.00 ± 2.52, 81.33 ± 0.41 and 76.67 ± 1.20) and 4.5% (11.00 ± 4.51, 85.33 ± 9.88 and 84.00 ± 8.50), respectively, after thawing. SL at 0% had the highest (P < 0.05) values for MDA, cholesterol and seminal arginase activity (1.10 ± 0.008 nmol/ml, 236.35 ± 4.08 mg/dl and 0.54 ± 3.3 E-3 units/mg protein, respectively). Our data suggest that 1.5% sunflower lecithin can be used in place of soy lecithin as a substitute for egg yolk during the cryopreservation of caprine semen.

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