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.

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.

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 A Comparison of Freezing Methods and Diluents Types on Post-Thaw Sperm Quality of Ram Sperm

2020 ◽  
Vol 7 (2) ◽  
pp. 235-241
Author(s):  
Pankaj Kumar Jha ◽  
M Golam Shahi Alam ◽  
Farida Yeasmin Bari
Keyword(s):  
Plasma Membrane ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Sperm Parameters ◽  
Plasma Membrane Integrity ◽  
One Step ◽  
Acrosome Integrity ◽  
Ram Sperm

The effect of freezing methods and diluents types on post-thaw sperm quality of Bangladeshi ram semen was studied. Two freezing methods and three diluents was tested as pooling effects (freezing methods or diluents) on post-thaw sperm parameters; sperm motility (SM), viability (SV), plasma membrane integrity (SPMI) and acrosome integrity (SAI), respectively. From selected ten rams, eight ejaculates were used for each freezing group (freezing methods × diluents). Semen samples were diluted by using two-steps for hand-made tris-based diluents (20% egg yolk): D1 (7% glycerol) and D2 (5% glycerol), and one-step dilution for commercial diluents: D3 (Triladyl®) at 35°C. After 4h of equilibration of temperature at 5°C, diluted semen samples was aspirated into 0.25 mL straws, and sealed. Straws were frozen in liquid nitrogen (LN2) vapour using two methods: F1 (manually in Styrofoam box, using three-steps method; +5°C to -80°C at -11.33°C/min, -80°C to -120°C at -26.66°C/min, and -120°C to -140°C at - 13.33°C/min) and F2 (programmable bio-freezer, using two-steps method; +5°C to -100°C at - 20°C/min and -100°C to -140°C at -10°C/min). Two semen straws from each batch were evaluated (37°C for 20 sec) for sperm parameters. In pool effects between freezing methods; SAI differed significantly (P < 0.001). The SM (56%) and SV (72%) were observed competitive. However, SPMI (67.58 ± 2.02%) and SAI (76.13 ± 1.42%) were higher in F1. Among diluents, SM (P < 0.006), SV (P < 0.008), SPMI (P < 0.012) and SAI (P < 0.019) differed significantly. The SM (61.25 ± 1.80%), SV (77.13 ± 1.47%), SPMI (68.31 ± 1.91%) and SAI (74.75 ± 1.64%) were highest in D3. In conclusion, the combination of manual freezing (three-steps) and handmade tris-based diluents (20% egg yolk, 5% glycerol) is suitable and sustainable method for cryopreservation of ram semen. Res. Agric., Livest. Fish.7(2): 235-241,  August 2020

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.

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 Sperm quality assessments for endangered razorback suckers Xyrauchen texanus

Reproduction ◽  
2011 ◽  
Vol 141 (1) ◽  
pp. 55-65 ◽  
Author(s):  
Jill A Jenkins ◽  
Bruce E Eilts ◽  
Amy M Guitreau ◽  
Chester R Figiel ◽  
Rassa O Draugelis-Dale ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Natural Populations ◽  
Reproductive Capacity ◽  
Dna Integrity ◽  
Computer Assisted ◽  
Progressive Motility ◽  
Isotonic Buffer ◽  
Quality Assessments ◽  
Sperm Motion

Flow cytometry (FCM) and computer-assisted sperm motion analysis (CASA) methods were developed and validated for use with endangered razorback suckersXyrauchen texanuscollected (n=64) during the 2006 spawning season. Sperm motility could be activated within osmolality ranges noted during milt collections (here 167–343 mOsm/kg). We hypothesized that sperm quality of milt collected into isoosmotic (302 mOsm/kg) or hyperosmotic (500 mOsm/kg) Hanks' balanced salt solution would not differ. Pre-freeze viabilities were similar between osmolalities (79%±6 (s.e.m.) and 76%±7); however, post-thaw values were greater in hyperosmotic buffer (27%±3 and 12%±2;P=0.0065), as was mitochondrial membrane potential (33%±4 and 13%±2;P=0.0048). Visual estimates of pre-freeze motility correlated with total (r=0.7589; range 23–82%) and progressive motility (r=0.7449) by CASA and were associated with greater viability (r=0.5985;P<0.0001). Count (FCM) was negatively correlated with post-thaw viability (r=−0.83;P=0.0116) and mitochondrial function (r=−0.91;P=0.0016). By FCM-based assessments of DNA integrity, whereby increased fluorochrome binding indicated more fragmentation, higher levels were negatively correlated with count (r=−0.77;P<0.0001) and pre-freeze viabilities (r=−0.66;P=0.0004). Fragmentation was higher in isotonic buffer (P=0.0234). To increase reproductive capacity of natural populations, the strategy and protocols developed can serve as a template for use with other imperiled fish species, biomonitoring, and genome banking.

scholarly journals PENGGANTIAN BOVINE SERUM ALBUMIN PADA PENGENCER CEP-2 DENGAN SERUM DARAH SAPI DAN PUTIH TELUR TERHADAP KUALITAS SEMEN CAIR SAPI LIMOUSIN SELAMA PENDINGINAN (The Substitution of Bovine Serum Albumin with Cattle Blood Serum and Egg White in CEP-2 Diluent on the Quality of Limousin Bull Liquid Semen during Refrigerating)

2016 ◽  
Vol 10 (2) ◽  
pp. 98-102
Author(s):  
Trinil Susilawati ◽  
Feri Eka Wahyudi ◽  
Inna Anggraeni ◽  
Nurul Isnaini ◽  
Muhammad Nur Ihsan
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Blood Serum ◽  
Sperm Motility ◽  
Bovine Serum ◽  
Sperm Quality ◽  
Egg Yolk ◽  
Egg White ◽  
Cattle Blood

This study aims to determine the effect of the substitution of bovine serum albumin (BSA) with cattle blood serum and egg white in the diluent of Cauda epididymal Plasma 2 (CEP-2) on sperm quality of Limousin cattle during cooling at 3-5 C. The research material used was rejected Limousin bull sperm (motility of 50-60%) from Artificial Insemination Centre Singosari, Malang. This research was a laboratory experiment using a randomized block design which was composed of six treatments with 10 replications, those were T0 as controls ((90% CEP-2 with BSA + 10% egg yolk); T1 (83.84% CEP-2 + 6.16% cattle blood serum + 10% egg yolk); T2 (81.84% CEP-2 + 8.16% cattle blood serum + 10% egg yolk); dan T3 (90% CEP-2 + 0,4% egg white + 10% egg yolk); T4 (90% CEP-2 + 0.8% egg white + 10% egg yolk); and T5 (90% CEP-2 without BSA + 10% egg yolk). Parameters measured were the percentage of motility, viability, and abnormality of sperms. Results of research after 48 hours of storage showed that the percentage of sperm motility in T0, T1, T2, T3, T4, and T5 were 40.50±5.90, 36±36.16, 34.00±6.58, 40.50±3.69, 38.50±3.37, and 38.50±4.12, respectively, while the percentage of sperms viability were 75.16±4.30, 70.50±2.88, 73.80±2.80, 74.80±3.30, 75.13±3.13, and 74.03±4.13, respectively, and the percentage of sperms abnormality were 10.14±2.34, 10.62±1.34, 11.33±2.00, 10.94±2.82, 10.02±1.95, and 10.78±1.96, respectively. In conclusion, CEP-2 diluent with or without the addition of 19% egg yolk in BSA and the substitution of BSA with 0.4-0.8% egg white can maintain semen quality to hour of 48 in cold storage.This study aims to determine the effect of the substitution of bovine serum albumin (BSA) with cattle blood serum and egg white in the diluent of Cauda epididymal Plasma 2 (CEP-2) on sperm quality of Limousin cattle during cooling at 3-5 C. The research material used was rejected Limousin bull sperm (motility of 50-60%) from Artificial Insemination Centre Singosari, Malang. This research was a laboratory experiment using a randomized block design which was composed of six treatments with 10 replications, those were T0 as controls ((90% CEP-2 with BSA + 10% egg yolk); T1 (83.84% CEP-2 + 6.16% cattle blood serum + 10% egg yolk); T2 (81.84% CEP-2 + 8.16% cattle blood serum + 10% egg yolk); dan T3 (90% CEP-2 + 0,4% egg white + 10% egg yolk); T4 (90% CEP-2 + 0.8% egg white + 10% egg yolk); and T5 (90% CEP-2 without BSA + 10% egg yolk). Parameters measured were the percentage of motility, viability, and abnormality of sperms. Results of research after 48 hours of storage showed that the percentage of sperm motility in T0, T1, T2, T3, T4, and T5 were 40.50±5.90, 36±36.16, 34.00±6.58, 40.50±3.69, 38.50±3.37, and 38.50±4.12, respectively, while the percentage of sperms viability were 75.16±4.30, 70.50±2.88, 73.80±2.80, 74.80±3.30, 75.13±3.13, and 74.03±4.13, respectively, and the percentage of sperms abnormality were 10.14±2.34, 10.62±1.34, 11.33±2.00, 10.94±2.82, 10.02±1.95, and 10.78±1.96, respectively. In conclusion, CEP-2 diluent with or without the addition of 19% egg yolk in BSA and the substitution of BSA with 0.4-0.8% egg white can maintain semen quality to hour of 48 in cold storage.

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
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