117. THE COMPARISON OF SPERM FREEZABILITY USING TWO EGG YOLK-FREE DILUENTS IN ZANDI RAM

2009 ◽  
Vol 21 (9) ◽  
pp. 36
Author(s):  
R. Ardebili ◽  
A. Towhidi ◽  
S. Zeinodini ◽  
M. Bahreini
Keyword(s):  
Recovery Rate ◽  
Sperm Quality ◽  
Calculated Data ◽  
Egg Yolk ◽  
Freezing Process ◽  
Automatic Equipment ◽  
Progressive Motility ◽  
Motility Of Sperm ◽  
Qualitative Characteristics

The aim of this study was to investigate the effect of two kinds of commercial egg yolk-free extenders: Bioxcell and AndroMed on in vitro sperm function of sheep. A total of 10 ejaculates were collected from 5 adult Iranian Zandi rams using artificial vagina. Semen samples were mixed and diluted with Bioxcell or AndroMed. Freezing was conducted using imv semi-automatic equipment. The percentage of motility and progressive motility of sperm were evaluated before freezing (at 37°C), after refrigeration (at 5°C) and thawing, and recovery rate was calculated. Data was analyzed using proc GLM of SAS. The effect of extender on sperm qualitative characteristics after thawing was significant. Mean percentage of motility, progressive motility and recovery rate were higher (P≤0.01) in AndroMed than Bioxcell. Results suggested that AndroMed in comparison with Bioxcell had more ability to preserve sperm quality during freezing process.

85 THE COMPARISON OF SPERM FREEZABILITY USING TWO EGG YOLK-FREE DILUENTS IN ZANDI RAM

2010 ◽  
Vol 22 (1) ◽  
pp. 201
Author(s):  
R. Ardebili ◽  
A. Towhidi ◽  
S. Zeinodini ◽  
M. Bahreini ◽  
E. Dirandeh
Keyword(s):  
Least Squares ◽  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Recovery Rate ◽  
Sperm Quality ◽  
Egg Yolk ◽  
Freezing And Thawing ◽  
Automatic Equipment ◽  
Progressive Motility ◽  
Motility Of Sperm

The aim of this study was to investigate the effect of 2 different commercial egg yolk-free extenders, Bioxcell and AndroMed, on post-thaw ram sperm motility and recovery rate. A total of 10 ejaculates were collected from 5 adult Iranian Zandi rams using artificial vagina during 3 weeks, once per week. Preexamination was conducted on semen, and semen motility >70% were selected. Semen samples were pooled and diluted with Bioxcell or AndroMed. Pooled semen was split into 2 treatments and extended to a final concentration of 200 × 108 spermatozoa per mL with extenders. The extended semen was manually packed into 0.25-mL mini-straws and sealed with PVC powder. After 4 h of equilibration in a waterbath at 5°C, the straws were dried with paper tissue, placed horizontally on a rack, and transferred to an isothermal box to be frozen in vapor 5 cm above liquid nitrogen. After 10 min, the straws were plunged into liquid nitrogen. Freezing was conducted using IMV semi-automatic equipment. The percentage of motility and progressive motility of sperm were evaluated before freezing (at 37°C), after refrigeration (at 5°C) and thawing, and recovery rate was calculated. To evaluate indices of sperm motility including the percentage of motile sperm and the progressive forward motility, semen was diluted with 0.9% NaCl w/v (1:100). Ten μL of diluted semen was placed on the prewarmed (37°C) microscope slide (76.2 mm × 24.5 mm; Pearl, China) covered with a cover slip (24 × 24 mm; Menzel-Glaser, Germany) and examined 10 different fields under a phase contrast microscope (Nikon, Tokyo, Japan), at ×200. Dilution factor 1:5 was used. The microscope was equipped with warm plate set at 37°C. Data were analyzed using Proc GLM of SAS. The effect of extender on sperm qualitative characteristics after thawing was significant. Least squares mean percent motility and progressive motility of AndroMed (36.05% ± 0.75; 29.0% ± 0.84) was significantly higher (P < 0.01) than Bioxcell (8.02% ± 0.35; 4.0% ± 0.60); least squares mean recovery rate in AndroMed (34.23% ± 1.05) also was higher (P < 0.01) than Bioxcell (10.0% ± 0.92). Results suggested that AndroMed in comparison with Bioxcell had more ability to preserve sperm quality during the freezing and thawing process.

P–126 A comparison of the efficacy of sperm freezing using high security tubes versus high security straws

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
A Bañuelo. Linares ◽  
K Berrisford ◽  
L Kellam ◽  
A Campbell
Keyword(s):  
Sperm Motility ◽  
Recovery Rate ◽  
Sperm Quality ◽  
Freezing Process ◽  
Donor Sperm ◽  
Progressive Motility ◽  
Sperm Density ◽  
High Security ◽  
Sperm Freezing ◽  
Sperm Recovery

Abstract Study question Are there any advantages in using High security tubes rather than High Security straws for conventional slow sperm freezing? Summary answer Freezing sperm in High Security tubes (HST) improved post-thaw recovery rate and motility, and also reduced processing and handling compared to High Security straws (HSS). What is known already The use of High Security freezing consumables (HSFC) in an IVF setting is a safe and effective way of eliminating concerns related to viral cross-contamination during storage. The lower diameter of HSS does make them susceptible to warming during handling. The HSFC used in this study is the only CE marked products that are made of resin, leak-proof and shatter-proof in all cryogenic temperatures even in LN2. No previous studies have compared the use of HST with HSS for conventional human sperm freezing. This study sets out to investigate the performance of HST compared to HSS. Study design, size, duration The study was designed as a controlled split-sample study with blind post-thaw analysis. Following the routine WHO analysis of 20 semen samples, the remainder of each of the samples was evenly divided and cryopreserved by conventional slow freezing in each of the two different HSFC. The freeze was conducted simultaneously by the same practitioner, employing the same freezing protocol and cryoprotectant. The pre-freeze and post-thaw concentration, total and progressive sperm motility were recorded. Participants/materials, setting, methods At one IVF clinic, semen samples with sperm density ≥15million/ml, ≥40% motility, ≥1.5ml were included. Cryoprotectant (SpermFreeze, Fertipro) was added dropwise to unprepared semen and kept at room temperature for 10 minutes before loading into HSFC (0.5ml CBS™HSS; CBS™HST). HSFC were heat-sealed (SYMS; SYMSIII sealers) and placed in vapour for 30 minutes before plunging into LN2. Samples were thawed by immersion in a 37Cº water bath for 5 minutes and analysed using WHO methods. Main results and the role of chance Paired-t test was used to compare the percentage motility between the different HSFC. All analysis was considered statistically significant when p &lt; 0.01. We demonstrated that the sperm recovery rate (Percentage total motility post-thaw/ Percentage total motility pre-freeze) in HST was 66.63 ± 14.94 (mean ± standard deviation) compared to 40.80 ± 14.69 in HSS. In the HSS, the percentage post-thaw total motility was 19.99 ± 7.21 and the percentage post-thaw progressive motility was 12.26 ± 2.59. In the HST, the percentage post-thaw total motility was 32.57 ± 8.33 and the percentage post-thaw progressive motility was 23.08 ± 5.53. The overall improvement when using HST against HSS was 12.53 ± 5.69, 10.44 ± 5.29 for the total motility and the progressive motility respectively. Comments were recorded regarding the handling and the condition of the HSS and HST for each freeze event. Neither device displayed any leakage of LN2 or any explosion during the warming. The freezing process was easier and faster using HST rather than HSS. It was also noted that the entire sample can be recovered from the HST, unlike the HSS. Limitations, reasons for caution The study looked at sperm recovery in terms of motility only. DNA damage was not considered as a parameter of sperm quality. Also, fertilization, pregnancy rates, live birth rates and the use of poorer quality sperm samples have not been investigated. Wider implications of the findings: For conventional sperm freezing, the use of HST resulted in improved sperm motility and progression post-thaw, when compared to HSS. This finding supports the use of HST to improve the post thaw quality of sperm, benefitting patients with own frozen samples, recipients of donor sperm and donor sperm banks. Trial registration number Not applicable.

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 Effect of fruit-juice on spermatozoa viability and lipid peroxidation of Red Sokoto bucks during liquid storage

2021 ◽  
Vol 41 (1) ◽  
pp. 16-27
Author(s):  
J. O. Daramola ◽  
T. A. Sorongbe ◽  
O. M. Onagbesan ◽  
A. V. Jegede ◽  
A. O. Ladokun ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Cell Damage ◽  
Egg Yolk ◽  
Protective Effects ◽  
Sperm Viability ◽  
Progressive Motility ◽  
Liquid Storage ◽  
Watermelon Juice ◽  
And Control

Antioxidants are linked with sperm viability because of their protective effects against cell damage during preservation. In order to enhance the life span of refrigerated buck semen, this study was carried out to determine the effect of fruit-rich antioxidants on spermatozoa viability and lipid peroxidation (LPO) of buck semen during liquid storage. Pooled semen from five Red Sokoto bucks was diluted with Tris-egg yolk based extender and supplemented each with juices from pawpaw tomato and watermelon at 0, 2.5, 5, 7.5 and 10/ 100 ml respectively. Following dilution, the semen samples were assessed subjectively after in vitro storage at 5°C for 24, 48, 72 and 96 hours as regards sperm motility, abnormalities, and acrosome status using a phase-contrast microscope. The concentration of malondialdehyde (MDA) as indices of lipid peroxidation (LPO) in the stored semen was measured in thiobarbituric acid reactive substances (TBARS) at 24, 48, 72 and 96 hours. The results showed highest progressive motility in watermelon juice at 2.5% (P<0.05) during the first 24 hours of storage while the lowest progressive motility was recorded at various levels of pawpaw juice (P<0.05). After 48 hours of storage, extender supplemented with watermelon and tomato juices had better progressive motility compared to control except 7.5% and 10%% of tomato juice (P<0.05). Irrespective of level of juice in the extender, the percentage of intact acrosome was similar among the various juices and control. The results showed that spermatozoa extended with watermelon juice had the lowest (P<0.05) percentage abnormality compared to other extenders at 24, 48, 72 and 96 hours of storage. Higher (P<0.05) percent spermatozoa abnormality compared to other fruit juices and control was observed at 72 and 96 hours of storage in spermatozoa extended with pawpaw juice. Significant reductions of MDA concentrations were achieved by addition of fruit-rich antioxidants to Tris-egg yolk based extender during the first 72 hours and the reduction was much pronounced in extender supplemented with pawpaw juice compared to control (P<0.05). The findings reveal that fruit-rich antioxidants from watermelon and tomato have protective ability to maintain sperm viability and to reduce concentration MDA of buck semen during liquid storage.

INFLUENCE OF GLYCEROL CONCENTRATION AND FREEZING TO −79 C ON OXYGEN UPTAKE AND LIVABILITY OF BOAR AND BULL SPERMATOZOA

1972 ◽  
Vol 52 (1) ◽  
pp. 65-72 ◽  
Author(s):  
L. M. SANFORD ◽  
G. J. KING ◽  
J. W. MACPHERSON
Keyword(s):  
Oxygen Uptake ◽  
Incubation Period ◽  
Skim Milk ◽  
Egg Yolk ◽  
Freezing Process ◽  
Glycerol Concentration ◽  
Motile Cells ◽  
Boar Semen ◽  
Boar Spermatozoa

Boar and bull spermatozoa were diluted in a skim milk–egg yolk–glucose extender containing 0, 7.5, or 15% glycerol (v/v) and incubated aerobically for 6 hr at 37 C. Other partially diluted boar semen samples were cooled to 5 C. Glycerol was added to a final concentration of 0, 7.5, and 15%. Samples were frozen to −79 C, rewarmed, and incubated for 3 hr at 37 C. The presence of glycerol in the extender depressed (P < 0.01) the oxygen uptake by nonfrozen boar and bull spermatozoa during the 6-hr incubation period. The reduction of oxygen uptake by semen samples increased as the level of glycerol in the extender increased. There was a corresponding decrease (P < 0.01) in the number of motile cells at the conclusion of the incubation period. Glycerol appeared to have more of a detrimental effect on boar spermatozoan oxygen uptake. The rate of oxygen uptake by boar semen samples postfreezing was extremely depressed, suggesting that spermatozoa surviving the freezing process metabolize at a much lower rate than normal. Active progressive motility of most of the surviving boar spermatozoa ceased within 1–2 hr of incubation under the in vitro conditions of this experiment.

The use of butylated hydroxytoluene in cryopreservation of boar semen

2016 ◽  
Vol 12 (2) ◽  
pp. 21-28
Author(s):  
Monika Trzcińska ◽  
Magdalena Baryła
Keyword(s):  
Dna Fragmentation ◽  
Sperm Quality ◽  
Annexin V ◽  
Egg Yolk ◽  
Butylated Hydroxytoluene ◽  
Preimplantation Embryos ◽  
Progressive Motility ◽  
Boar Semen ◽  
Boar Spermatozoa

The objective of the study was to determine the effect of butylated hydroxytoluene (BHT) on the quality and fertilizing capacity of frozen-thawed (FT) boar semen. Semen from five boars (36 ejaculates) was resuspended in lactose-egg yolk-glycerol extender supplemented with 0 (control), 1.0 (R1), 1.5 (R2) or 2.0 mM BHT (R3). Sperm quality was assessed based on motility (CASA; TM: total motility; PM: progressive motility), phosphatidylserine (PS) translocation across the plasma membrane (Annexin-V-FLuos Staining Kit) and DNA fragmentation (TUNEL Assay). The FT semen was also used for intrauterine artificial insemination (AI) of synchronized gilts. The fertilizing capacity of the FT semen was assessed on the basis of the gilt insemination rate and the number of morphologically normal embryos. The quality of the preimplantation embryos was determined by observing a TUNEL-positive reaction. The highest percentage of progressive motile and viable spermatozoa was noted in extender R3 (74.8 ±4.4% and 63.7 ±5.8%), as compared with the control (38.3 ±2.8% and 36.1 ±2.6%). The addition of BHT to the extender did not increase early apoptotic changes in the frozen-thawed spermatozoa with respect to the control. Irrespective of the variant of the extender, cryopreservation and thawing did not induce fragmentation in the boar spermatozoa. The highest number of morphologically normal embryos from inseminated gilts was observed in the case of semen cryopreserved in extender supplemented with 1.5 mM BHT. No significant differences were observed in DNA fragmentation in the expanded blastocysts from gilts inseminated with FT semen cryopreserved in the extenders analysed.

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

2015 ◽  
Vol 55 (4) ◽  
pp. 551 ◽  
Author(s):  
M. Emamverdi ◽  
M. Zhandi ◽  
A. Zare Shahneh ◽  
M. Sharafi ◽  
A. Akhlaghi ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Soybean Lecithin ◽  
Sperm Quality ◽  
Egg Yolk ◽  
Mitochondrial Activity ◽  
Flow Cytometric ◽  
Microscopic Evaluation ◽  
Open Question ◽  
Ram Sperm

The present study was designed to determine the effect of three different extenders on ram sperm quality during a freeze–thawing procedure using flow cytometric and microscopic evaluations. Several in vitro qualitative analyses of post-thawed sperm parameters including motility and velocity parameters, plasma membrane functionality, total abnormality, capacitation status, acrosome integrity, mitochondrial activity and apoptosis features were considered. In the breeding season, seven ejaculates from each Zandi ram were collected routinely twice a week. Following semen collection, samples were pooled and equally divided into three aliquots. Each aliquot was diluted and frozen with one of the following extenders: (1) Tris-based extender containing 1.5% (w/v) soybean lecithin (TSL), as a chemically defined extender, (2) Bioxcell, a commercial soybean lecithin-based extender, and (3) Tris-based extender containing 20% (v/v) egg yolk (TEY). The results of the present study indicated no differences in total [TSL (55.8 ± 2.02%) vs TEY (50.2 ± 2.02%; P < 0.05)] and progressive motility of spermatozoa [TSL (26.2 ± 1.36%) vs Bioxcell (22.4 ± 1.36%; P < 0.05)]. Semen freezing by means of TSL resulted in a higher percentage of live spermatozoa (39.42 ± 1.81%) compared with TEY (29.17 ± 1.81%; P < 0.05), and a higher percentage of functional plasma membrane (50.8 ± 192%) compared with TEY (44 ± 1.92%) and Bioxcell (38.8 ± 1.92%; P < 0.05). The effect of extenders on sperm capacitation status showed that the percentage of post-thawed capacitated spermatozoa was higher in TEY (61.9 ± 1.48%) compared with that in TSL (56.6 ± 1.48%; P < 0.05). The evaluation of post-thawed spermatozoa indicated that the percentage of live spermatozoa with active mitochondria was higher in TSL (53.05 ± 2.31%) compared with Bioxcell (45.92 ± 2.31; P < 0.05) and the percentage of intact acrosome spermatozoa was higher in TSL (84.55 ± 2.51%) compared with TEY (74.91 ± 2.51%; P < 0.05). The use of TSL and Bioxcell extenders reduced the percentage of apoptotic spermatozoa (40.82 ± 2.07% and 42.22 ± 2.07%, respectively), compared with TEY (51.34 ± 2.07%; P < 0.05). Post-thawing dead spermatozoa were increased when semen was frozen by Bioxcell (25.69 ± 1.28%). The results of this study showed that TSL extender may provide stabile milieu and conditions for ram sperm cryopreservation compared with Bioxcell and TEY extenders. Whether TSL extender can improve the artificial insemination results remains, however, an open question.

scholarly journals Effect of whole and clarified egg yolk based extenders on post-thaw semen quality in bulls.

2021 ◽  
Author(s):  
Prosper Kamusasa ◽  
Eddington Gororo ◽  
Fungayi Primrose Chatiza
Keyword(s):  
Semen Quality ◽  
Sperm Quality ◽  
Egg Yolk ◽  
Quality Parameters ◽  
Normal Morphology ◽  
Semen Cryopreservation ◽  
Bull Semen ◽  
Inclusion Level ◽  
Whole Egg

Abstract This study was conducted to evaluate the comparative cryoprotective effects of whole egg yolk and clarified egg yolk on post thaw sperm quality parameters and to determine the optimum clarified egg yolk inclusion level (10-20%) in semen extenders for Mashona bull semen cryopreservation. It was shown that there was a significant decrease in sperm quality variables following cryopreservation. Semen quality increased with the concentration of clarified egg yolk, indicating a positive relationship between egg yolk LDL concentration and maintenance of in vitro sperm quality. The 20% clarified egg yolk (CEY20) extender treatment gave post-thaw motility, viability and normal morphology values which were comparable to the control (20% whole egg yolk, WEY20). The 10% clarified egg yolk concentration gave the least post-thaw quality values and the greatest proportion of defective spermatozoa. This experiment found no advantage of replacing whole egg yolk with up to 15% clarified egg yolk in Mashona bull semen cryopreservation. However, 20% clarified and 20% whole egg yolk performed similarly in the maintenance of post-thaw sperm motility, viability and normal morphology.

scholarly journals Effects of season, breed and age on the boar sperm quality: 2. The extended sperm

2003 ◽  
Vol 19 (3-4) ◽  
pp. 25-29
Author(s):  
Blagoje Stancic ◽  
Mladen Gagrcin ◽  
Ivan Radovic
Keyword(s):  
Sperm Quality ◽  
Progressive Motility ◽  
Boar Sperm ◽  
June July

Total of 182 ejaculates were extended in 1:4 proportions and investigated for progressive motility (PM) ratio, after 24h and 72h of storage at +170C. The boars of Yorkshire, Landrace, Duroc, Hempshire and Pietrain pure breeds and F1-crossbreeds were divided according to age as: ?12, 13-18, 19-24 and ?25 months. Extended sperm samples were investigated within 4 seasons of year January-March, April-June, July-September and October-December. The PM ?65% was estimated in 60% of ejaculates after 24h and in 25% ejaculates after 72h preservation in vitro. Some differences in progressive motility, influenced by season, breed and age of boars, were established. Present results are preliminary and require further investigations.

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