COMPARATIVE CHARACTERISTICS OF RAM SEMEN FROZEN IN DIFFERENT EXTENDERS

2021 ◽  
pp. 63-78
Author(s):  
M.M. AYBAZOV ◽  
◽  
A.N. SHEVCHENKO ◽  
M.I. SELIONOVA ◽  
T.V. MAMONTOVA
Keyword(s):  
Egg Yolk ◽  
Quality Parameters ◽  
Semen Parameters ◽  
Progressive Motility ◽  
Kinematic Performance ◽  
Synthetic Media ◽  
Ram Sperm

Numerous studies have proved the necessity of egg yolk in synthetic media to dilute semen before its cryopreservation. However, at the same time, it has been demonstrated that its use can adversely affect the quality of frozen-thawed sperm. The present study compares the main quality parameters of ram sperm frozen using TRIS-based diluent with egg yolk and two egg yolk-free diluents (OvixCell® and AndroMed®). A slower deterioration in the kinematic performance of sperm cryopreserved in TRIS diluent with native egg yolk confirmed higher cryoprotective performance compared to commercial extenders containing no egg yolk. Significantly higher total and progressive motility was observed in TRIS-based medium with egg yolk (P< 0.05). This advantage was maintained after four hours of incubation and became more significant at the end of cultivation (after six hours) (P< 0.01). Thus, ram sperm frozen in egg yolk medium retained better motility than in egg yolk-free extenders, which allows predicting its higher bioavailability. Assessment of some semen parameters using CASA showed that there were no significant differences in motility between the three extenders immediately after thawing the straws. When assessed two hours after thawing, a diluent containing egg yolk (TRIS-based) was found to have higher results for some of the examined traits than phospholipid diluents.

scholarly journals THE PRODUCTION OF FREEZE-DRIED EGG YOLK POWDER AND ITS EFFECT ON THE QUALITY OF GARUT RAM LIQUID SEMEN

2021 ◽  
Vol 15 (2) ◽  
Author(s):  
Oriza Savitri Ariantie ◽  
Amrozi Amrozi ◽  
Tuty Laswardi Yusuf ◽  
Nurul Taufiqu Rochman ◽  
Bambang Purwantara
Keyword(s):  
Particle Size ◽  
Sperm Quality ◽  
Egg Yolk ◽  
Progressive Motility ◽  
Freeze Dried ◽  
Semen Preservation ◽  
Viability Percentage ◽  
Index Value ◽  
Ram Sperm

This research aims to evaluate the protective potency of freeze-dried egg yolk powder on Garut ram sperm during the liquid semen preservation process. Semen with good sperm quality was divided into three groups and diluted using the following Tris diluents: fresh egg yolk-Tris (FEY-Tris), commercial egg yolk powder-Tris (CEY-Tris), and freeze-dried egg yolk powder-Tris (DEY-Tris). Semen that had been diluted was observed every 12 hours until sperm progressive motility was 50%. Sperm quality was tested for progressive motility percentages, viability, intact plasma membrane (IPM), and intact acrosome (IA). Fresh egg yolk had a particle size of 14460.00±330.76 nm, with polydispersity index value (PI) and zeta potential (ZP) being 1.00±0.11 and -9.22±0.30 mV, respectively. The particle size of commercial egg yolk powder was 877.90±168.86 nm with PI value 0.34±0.04 and ZP-28.7±1.24 mV. The particle size of freeze-dried egg yolk powder was 1296.00±86.73 nm, with PI value 0.70±0.04 and ZP -34.5±0.64 mV. Progressive motility percentage showed that DEY-Tris diluent managed to survive to 51.11±6.06% for 168 hours, compared to FEY-Tris (51.25±6.74%) which survived for 156 hours, although no longer than CEY-Tris diluent (53.47±6.33%) which survive for 180 hours (P0.05). Higher sperm motility was also supported by viability percentage, IPM, and IA, which were all higher (P0.05). In conclusion, freeze dried egg yolk powder mixed with Tris buffer was able to maintain sperm quality during the liquid semen preservation process.

Value of semen parameters, with special reference to TNF-α, in predicting the quality of boar semen after short-term storage

2013 ◽  
Vol 61 (2) ◽  
pp. 209-219 ◽  
Author(s):  
Janko Mrkun ◽  
Marjan Kosec ◽  
Petra Zrimšek
Keyword(s):  
Semen Quality ◽  
Semen Parameters ◽  
Short Term ◽  
Progressive Motility ◽  
Tnf Α ◽  
Semen Storage ◽  
Boar Semen ◽  
Α Level ◽  
Term Storage

The aim of this study was to address the question whether changes in boar semen quality after short-term storage could be predicted on the basis of standard semen parameters and TNF-α level determined on the day of semen collection under commercial conditions. Progressive motility showed the highest positive correlation with morphology on day 0 of collection, and progressive motility on day 3 (P < 0.05) showed a negative correlation with acrosome abnormalities (P < 0.05). According to the area under receiver operating characteristics (ROC) curves (AUCs), progressive motility could also be used in predicting semen quality after 3 days of storage (AUC > 0.5; P < 0.05). TNF-α in seminal plasma is the only parameter measured on day 0 to show a significant correlation with the percentage of viable spermatozoa after 3 days of semen storage (r = 0.495, P < 0.05). ROC analysis shows that TNF-α level is helpful in discriminating viability outcome after semen storage (AUC = 0.94, P < 0.001). We can predict with 92.35% certainty that fresh semen samples with more than 150 pg/ml of TNF-α in the seminal plasma will retain more than 85% of viable spermatozoa after 3 days of storage. Thus, TNF-α can contribute to predicting the quality of short-term stored semen.

scholarly journals Cholesterol-loaded cyclodextrin plus trehalose improves quality of frozen-thawed ram sperm

2019 ◽  
Vol 64 (No. 03) ◽  
pp. 118-124 ◽  
Author(s):  
ME Inanc ◽  
S Gungor ◽  
C Ozturk ◽  
F Korkmaz ◽  
I Bastan ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Mitochondrial Membrane ◽  
Improve Quality ◽  
Progressive Motility ◽  
Computer Aided ◽  
Acrosome Integrity ◽  
Ram Sperm ◽  
Additive Control ◽  
Artificial Vagina

The objective of this study was to determine effects of supplementing Tris-based semen extenders with either cholesterol-loaded cyclodextrin (CLC) or 7-dehydrocholesterol loaded cyclodextrin (7-DCLC) plus trehalose (T) for cryopreservation of ram semen. Semen was collected with an artificial vagina from five Merino rams (2–3 years of age) during the non-breeding season. Ejaculates were pooled, divided into eight equal portions, diluted with a standard Tris-based extender containing: no additive (control); T (50 mM); or T (50 mM) + 1.5, 2.5 or 3.5 mg of either 7-DCLC or CLC. Semen was chilled from 37°C to 4°C, placed in 0.25 ml French straws, held 5 cm above liquid nitrogen for 12 minutes, then plunged into liquid nitrogen. After thawing, a computer-aided semen analyzer system (CASA) was used to assess motility, whereas plasma membrane and acrosome integrity (PMAI) and high mitochondrial membrane potential (HMMP) were assessed with flow cytometry. Sperm supplemented with 2.5 mg and 3.5 mg CLC + T had the highest (P &lt; 0.05) total and progressive motility (65.2 ± 4.7 and 19.0 ± 1.0% respectively, mean ± SEM), albeit with no significant differences from sperm with 1.5 or 3.5 mg CLC + T. Sperm with 2.5 mg CLC + T had the highest (P &lt; 0.05) PMAI (59.3%; not different from 3.5 mg CLC + T) and highest (P &lt; 0.05) HMMP (64.6%; not different from 1.5 or 3.5 mg CLC + T). The lowest ALH value, 2.8 ± 0.3 µm was in the 2.5 mg 7-DCLC + T group; otherwise, there were no significant differences among groups for any other CASA end point. In conclusion, adding CLC + T to a tris-based extender optimized quality of frozen-thawed ram semen. Therefore, extenders including CLC + T have potential to improve quality of frozen-thawed ram sperm.

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.

scholarly journals Effect of two freezing extenders on characteristic of fresh and frozen-thawed semen in endangered Old Kladruber stallions – A pilot study

2017 ◽  
Vol 62 (No. 6) ◽  
pp. 227-233 ◽  
Author(s):  
J. Šichtař ◽  
A. Nehasilová ◽  
O. Šimoník ◽  
F. Bubeníčková
Keyword(s):  
Egg Yolk ◽  
Quality Parameters ◽  
Computer Assisted ◽  
Kinematic Parameters ◽  
Fluorescence Staining ◽  
Sperm Analysis ◽  
Sperm Characteristic ◽  
Computer Assisted Sperm Analysis ◽  
Fresh Semen

The aim of the study was to evaluate the effect of two different extenders on sperm characteristic before equilibration and post-thaw in the endangered Old Kladruber stallions. Also, the response of individual stallions to the extenders used was tested. Semen was collected from six stallions every other day within one week. After centrifugation of the collected sperm-rich fraction, the supernatant was removed and sperm pellets were divided to two aliquots; these were diluted either with Gent (Minitube, Germany) or privately manufactured lactose-EDTA-egg yolk extender (Lact). Three cryopreserved insemination doses (IDs) from each extender (Gent and Lact) were prepared for each stallion from one collection (108 samples from six stallions in total). As a parameter of quality, the motility (computer assisted sperm analysis), viability (fluorescence staining), and morphology (eosin/nigrosine staining) were evaluated after dilution with freezing extenders (fresh) and after thawing (frozen-thawed). The different effects of chosen extenders on the quality of fresh semen were only manifested in higher kinematic parameters of sperm when the Lact extender was used. However, in frozen-thawed samples, the Gent extender yielded significantly better results in all of the evaluated parameters. The representation of sperm subpopulation was significantly influenced by extender in fresh as well as frozen-thawed samples; moreover, we found a significant effect of freezing on the distribution of these subpopulations. The response of individual stallions to chosen extenders was evident in the different quality of fresh as well as frozen-thawed IDs; Gent extender yielded better frozen-thawed IDs. Based on our results, among others describing quality parameters of ejaculate in endangered Old Kladruber stallions, we can recommend using Gent extender for the production of frozen-thawed IDs.  

scholarly journals The Use of κ-Carrageenan in Egg Yolk Free Extender Improves the Efficiency of Canine Semen Cryopreservation

Animals ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 88
Author(s):  
Eunji Kim ◽  
Areeg Almubarak ◽  
Nabeel Talha ◽  
Il-Jeoung Yu ◽  
Yubyeol Jeon
Keyword(s):  
Egg Yolk ◽  
Control Group ◽  
Spermine Oxidase ◽  
Plant Polysaccharide ◽  
Red Seaweeds ◽  
Spermine Synthase ◽  
Progressive Motility ◽  
Overall Efficiency ◽  
Lower Expression

κ-Carrageenan is a plant polysaccharide derived from red seaweeds reported to possess potential medicinal and antioxidants activities. The present study aimed to identify the cryoprotective effects of κ-carrageenan on the quality of frozen-thawed canine semen. Twenty-eight ejaculates were collected and diluted in a Tris egg-yolk-free extender supplemented with various concentrations of κ-carrageenan (0.0%, 0.1%, 0.2%, 0.3%, and 0.5%). The addition of κ-carrageenan to the extender at a 0.2% concentration induced a significant increase in the total motility (TM) and the rapid progressive motility (RPM) of canine sperm. Among the experimental groups, the highest percentage of sperms with intact acrosomes was found in the 0.5% κ-carrageenan group (p < 0.05). Apoptosis levels were significantly lower in the 0.1% and 0.2% κ-carrageenan treatment. Moreover, sperm in the κ-carrageenan supplemented group showed a significantly higher expression of antiapoptotic (Bcl-2) and lower expression of NADPH oxidase (NOX5), spermine synthase (SMS), and spermine oxidase (SMOX) genes than those in the control group. In conclusion, the addition of κ-carrageenan to the freezing extender improved the overall efficiency of frozen-thawed dog spermatozoa.

Effect of different extenders for preservation of ram semen at 4°C

2016 ◽  
Author(s):  
Haneef A. Rather ◽  
Rafiqul Islam ◽  
Asloob A. Malik ◽  
Farooz A. Lone ◽  
Mohamad Naiem Banday
Keyword(s):  
Citric Acid ◽  
Sperm Motility ◽  
Egg Yolk ◽  
Morphological Abnormalities ◽  
Progressive Motility ◽  
Hypoosmotic Swelling Test ◽  
Live Sperm

The aim of this study was to investigate the effects of different extenders viz. Tris citric acid fructose egg yolk (TCFEY), Tris citric acid glucose egg yolk (TCGEY), Egg yolk citrate fructose (EYCF) and Egg yolk citrate glucose (EYCG) on the quality of ram spermatozoa during preservation at 4°C. Semen samples showing more than 3+ mass motility and 70% progressive motility were pooled and subsequently divided into four aliquots. Each aliquot was extended separately in four different extenders viz. TCFEY, TCGEY, EYCF and EYCG and stored at 4°C up to 72h. The quality of spermatozoa on the basis of percentage of sperm motility, live sperm, morphological abnormalities, intact acrosome and hypoosmotic swelling test (HOST) reacted spermatozoa was evaluated immediately after extension in particular extenders (0 h), 24 h, 48 h and 72 h after preservation at 4°C. The percent sperm motility was significantly (P<0.01) higher for TCFEY and TCGEY than EYCF and EYCG at 72 h of preservation at 4°C. The percent HOST reacted spermatozoa and intact acrosomes were significantly (P<0.01) higher and morphological abnormalities were significantly (P<0.01) lower for Tris based fructose extender than other three extenders at 72 h at 4°C. In conclusion, Tris citric acid fructose egg yolk (TCFEY) was found the best in maintaining the quality of ejaculated ram spermatozoa during preservation for 72 h at 4°C. 

scholarly journals Trehalose addition to a Tris-fructose egg yolk extender on quality of ram sperm preserved at 0 °C

2020 ◽  
Vol 49 ◽  
Author(s):  
Jianqing Zhao ◽  
Guoliang Xiao ◽  
Wenliang Zhu ◽  
Di Fang ◽  
Na Li ◽  
...  
Keyword(s):  
Egg Yolk ◽  
Ram Sperm

scholarly journals Ram semen preserved at 0°C with soybean lecithin Tris-based extender substituted for egg yolk

2021 ◽  
Vol 34 (2) ◽  
pp. 192-197
Author(s):  
Jian-qing Zhao ◽  
Guo-liang Xiao ◽  
Wen-liang Zhu ◽  
Di Fang ◽  
Na Li ◽  
...  
Keyword(s):  
Soybean Lecithin ◽  
Egg Yolk ◽  
Control Group ◽  
Progressive Motility ◽  
Significant Difference ◽  
Acrosome Integrity ◽  
Normal Offspring ◽  
And Control ◽  
Ram Sperm ◽  
Artificial Vagina

Objective: The present study evaluated the preservation of ram semen at 0°C using soybean lecithin with a Tris-fructose extender.Methods: Semen was collected by artificial vagina ejaculation from six rams with proven fertility. High quality ejaculates were diluted by soybean lecithin (0.25%, 0.5%, 0.75%, 1.0%, 1.25%) using Tris-fructose extender and control (Tris-fructose egg yolk extender), respectively. The ejaculates were diluted to a concentration of 5×10<sup>8</sup> sperm/mL, followed by cooling to 0°C in 90 min and maintaining the temperature for 12 days. The diluted semen samples were examined and recorded for sperm progressive motility, acrosome integrity at 0, 24, 72, 144, 216, 288 h, respectively. Two hundred and twenty-three ewes were inseminated for 216 h with optimal soybean lecithin concentrated semen or control via trans-cervical insemination.Results: The results showed that there were no differences in sperm progressive motility at 0, 24, 72, and 144 h (p>0.05). After 216 h, the sperm progressive motility in the control group and 0.5% concentration groups was significantly higher when compared to 0.25% concentration (p<0.05). The 0.5% concentration group demonstrated the highest survival rate and had no difference with the control group (p>0.05). At 216 h, the sperm progressive motility of all groups was still above 50%. The acrosome integrity of all groups was decreased with prolongation of storage time, but there was no difference at each time point (p>0.05). There was no significant difference in the lambing rate and pregnancy rate between the 0.5% concentration group and the control group (p>0.05).Conclusion: These results suggest that ram sperm is capable of fertilization after preservation at 0°C with 0.5% of soybean lecithin in Tris-based extender substituted for egg yolk and produce normal offspring after insemination.

scholarly journals KUALITAS SPERMATOZOA AYAM PERANAKAN SENTUL DALAM PENGENCER RINGER LAKTAT KUNING TELUR DENGAN BERBAGAI MONOSAKARIDA (Quality of Sentul Crossbreed Chicken Spermatozoa in Ringer Lactate-Egg Yolk Diluents Supplemented with Various Monosaccharide)

2016 ◽  
Vol 10 (2) ◽  
pp. 166-169
Author(s):  
Khaeruddin Khaeruddin ◽  
Raden Iis Arifiantini ◽  
Cece Sumantri ◽  
Sri Darwati
Keyword(s):  
Energy Source ◽  
Semen Quality ◽  
Egg Yolk ◽  
Quality Parameters ◽  
Ringer Lactate ◽  
Spermatozoa Motility ◽  
Significant Difference

The aim of this study was to examine the preservation of sentul crossbreed chicken semen in ringer lactate egg yolk diluent supplemented with various monosaccharide. Semen was collected from three roosters using massage method. Immediately after collection, the semen was evaluated macroscopically and microscopically. Semen with more than 70% motility was divided into four tubes. Each of them diluted with ringer lactate egg yolk glucose (RLEYG), ringer lactate egg yolk fructose (RLEYF), ringer lactate egg yolk xylose (RLEYX) and ringer lactate egg yolk mannose (RLEYM). Semen was stored in refrigerator (5o C) for sixty hours and evaluated every twelve hours for spermatozoa motility and viability. Results showed that no significant difference (P>0.05) among diluents used on spermatozoa quality parameters after dilution and during preservation. Semen quality decrease during storage and at sixty hours of storage, the motility and viability of spermatozoa ranging from 48.33±2.56 to 55.42±2.26% and 58.59±2.87 to 64.83±2.42%, respectively. This research conclude that glucose, fructose, xylose and mannose can be used as energy source for roosters semen during preservation.

Sign in / Sign up

Export Citation Format

Share Document