scholarly journals THE EFFECT OF GLYCEROL CONCENTRATION IN TRIS GLUCOSE EGG YOLK EXTENDER ON THE QUALITY OF TIMOR DEER FROZEN SEMEN

2011 ◽  
Vol 36 (2) ◽  
Author(s):  
W.M.M. Nalley ◽  
R. Handarini ◽  
T.L. Yusuf ◽  
B. Purwantara ◽  
G. Semiadi
Keyword(s):  
Egg Yolk ◽  
Glycerol Concentration ◽  
Frozen Semen

Kualitas Semen Cair Kambing Boer Berbahan Pengencer Air Kelapa Muda Varietas Viridis Setelah Simpan Dingin

2019 ◽  
Vol 6 (1) ◽  
pp. 78
Author(s):  
Muhammad Ade Salim ◽  
Muhammad Nur Ihsan ◽  
Nur Isnaini ◽  
Trinil Susilawati
Keyword(s):  
Semen Quality ◽  
Block Design ◽  
Animal Husbandry ◽  
Egg Yolk ◽  
Coconut Water ◽  
Boer Goat ◽  
Randomized Block Design ◽  
Frozen Semen ◽  
Artificial Vagina

ABSTRAKAir kelapa muda varietas viridisdapat dijadikan pengencer aletrnatif semen cair bagi program IB di daerah minim sarana semen beku. Tujuan penelitian ini untuk menguji pengaruh penggunaan air kelapa muda viridissebagai bahan pengencer terhadap kualitas semen cair kambing Boer setelah didinginkan. Dilaksanakanselama 3 bulan di Laboratorium Fakultas Peternakan UBUnit SumberSekar,Malang. Metodenya yaitu eksperimen. Semen dari  3 pejantan Boer umur 3-5 tahun, dikoleksi seminggu sekali dengan VB. Air kelapa mudaviridis umur 5-7 bulan serta tris aminomethane sebagai kontrol. Didesain menggunakan Rancangan Acak Kelompok (RAK) dengan 2 perlakuan yaitu P0 (tris aminomethane + 10% KT) dan  P1 (air kelapa muda viridis + 10% KT) masing-masing diulang 10 kali. Data dianalisis dengan analisis Ragam (Anova) dengan software Genstat 18. Variabelnya yaitu motilitas individu, viabilitas dan abnormalitas. Hasil penelitian yaitu motilitas individu pada P1bertahan sampai 4 hari (40,5± 24,3%), viabilitas terbaik sampai hari ke-5 (42±24,6%), abnormalitas terendah di hari ke-7(1,31± 0,6). Kesimpulannya, Pengencer air kelapa muda viridis dapat mempertahankan kualitas semen cair kambing Boer selama 4 hari untuk motilitas dan 5 hari untuk viabilitas.Kata Kunci:pengencer, air kelapa, varietas viridisABSTRACTYoung viridis coconut water could be used as an alternative to liquid semen diluent for artificial insemination program in the area with limited facility for frozen semen production. This study evaluated the use of young coconut water as a diluent on liquid semen quality of Boer goat after cold storage. This study was carried out for 3 months at Sumber Sekar Laboratory, Faculty of Animal Husbandry, University of Brawijaya, Malang. The semen was collected from 3 Boer bucks aged at 3 to 5 years old. The semen collection was done once a week with the aid of artificial vagina. The diluents used were young Viridis coconut (5 to 7 months old) and tris aminomethane. The method used was an experiment in a randomized block design with 2 treatments and 10 replicates. The treatments used were T0: tris aminomethane + 10% egg yolk (control) and T1:  young Viridis coconut water + 10% egg yolk. Data were analyzed by analysis of variance using Genstat 18 software. The variables measured were sperm individual motility, viability, and abnormality. The results showed that the sperm individual motility in T1 survived up to 4 days (40.5± 24.3%), the best viability at 5 days (42.0±24.6%),  while the lowest abnormality at 7 days (1.31±0.6). It could be concluded that: 1. Tris aminomethane diluent has higher quality with the storage length up to 9 days, 2. Young Viridis coconut water diluent could preserve liquid semen quality of Boer goat up to 4 days for sperm motility and 5 days for sperm viability.Keywords: diluents, coconut water, viridis variety

76 THE EFFECT OF GLYCEROL CONCENTRATIONS IN EGG-YOLK CITRATE EXTENDER ON THE QUALITY OF CRYOPRESERVED NGUNI BULL SEMEN

2013 ◽  
Vol 25 (1) ◽  
pp. 185
Author(s):  
M. M. Seshoka ◽  
M. L. Mphaphathi ◽  
F. V. Ramukhithi ◽  
T. R. Netshirovha ◽  
C. Hlungwani ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Rural Areas ◽  
Egg Yolk ◽  
Positive Control ◽  
Negative Control ◽  
Glycerol Concentration ◽  
Bull Semen ◽  
Frozen Semen ◽  
Thawing Process ◽  
Motility Rate

There are bull shortages in South African poor rural areas. Artificial-insemination technology could play a significant role on breeding emerging farmer’s cattle. The objective of this study was to compare glycerol concentrations (0, 4, 8, or 12%) during freezing of Nguni bull semen to conduct AI in different villages. Semen was collected by electro ejaculator from 2 Nguni bulls of known and proven fertility. Collected semen samples were kept in a thermo flask (37°C) and transported to the laboratories within 10 min after collections. Semen samples were pooled and evaluated by Sperm Class Analyser® and allocated randomly per treatment. Semen was then diluted (1 : 2 v:v) with egg-yolk citrate extender supplemented with either 0% (negative control), 4, 8, and 12% of glycerol concentration or AndroMed® (positive control). Semen samples were equilibrated for 4 h at 5°C. After equilibration period, samples were loaded into 0.5-mL straws and placed horizontally into the controlled rate (–5, –8, –10, –12, –15, –25, –35°C min–1) from 5°C until target temperature of –80°C is reached. The frozen semen straws were stored in a liquid nitrogen tank (–196°C) until thawing. Treatment means were separated using Fisher’s protected t-test least, and data are presented as mean ± SD. There was a significant differences (P < 0.05) between raw total sperm motility (83.3 ± 19.3) and frozen–thawed sperm with either 0% (0.0 ± 0.0), 4% (30.2 ± 13.4), 8% (47.9 ± 12.5), or 12% (61.5 ± 4.7) of glycerol and on AndroMed® (27.7 ± 17.8) group. Regardless of the glycerol concentrations used, the freezing-thawing process reduced (P < 0.05) the Nguni total sperm motility rate compared to uncryopreserved sperm (83.3 ± 19.3). In conclusion, egg-yolk citrate extender supplemented with 12% glycerol yielded a better (P < 0.05) total sperm motility rate (61.5 ± 4.7) as compared with the 0% (0.0 ± 0.0), 4% (30.2 ± 13.4), 8% (47.9 ± 12.5), and AndroMed® (27.7 ± 17.8) group. Further studies are required to test other levels of glycerol concentrations (>12%) on freezing Nguni semen and conducting AI.

scholarly journals Effect of preservation time on the quality of frozen semen in indigenous rams

2015 ◽  
Vol 44 (1) ◽  
pp. 10-15 ◽  
Author(s):  
BBA Mahmuda ◽  
Azizun Nesa ◽  
BF Zohara ◽  
MGS Alam ◽  
FY Bari
Keyword(s):  
Semen Quality ◽  
Egg Yolk ◽  
Volume Density ◽  
Normal Morphology ◽  
Mean Values ◽  
Frozen Semen ◽  
Significant Difference ◽  
The Mean ◽  
Fresh Semen

The study was carried out to observe the effects of preservation time on the quality of frozen semen of indigenous rams. Semen was collected using AV once a week from 4 rams. Tris based with 10% egg yolk and 7% glycerol extender was used to extend and freezing the semen. Fresh semen was evaluated for volume, density, mass motility and concentration, and mean values were observed as 0.8±0.2ml, 3.0±0.3, 3.2±0.7, 3.9±0.7×109/ml, respectively. Significant difference (p<0.05) was found for all the parameters among the rams. Mean values of motility, viability and normal morphology percentages were 83.3±4.3%, 88.2±4.4%, 84.2±3.5% in fresh semen while those of chilled semen at 40C were 74.7±2.3, 78.8±4.9 and 79.2±2.9%, respectively. For all the parameters, significant (p<0.05) difference was found among the rams. Frozen sperm motility was observed after thawing at 39-400C for 14-15 seconds. The mean motility, viability and normal morphology percentages after freezing for 24hrs, 7, 15 and 30 days of duration were 39.8±3.1, 41.1±4.3, 40.1±4.1 and 39.4±2.9%; 44.5±2.5, 45.3±2.8, 44.6±2.8 and 43.9±2.8%; 71.0±2.0, 71.7±1.5, 70.7±1.7 and 70.3±1.8%, respectively and values did not decrease significantly (p>0.05) with the increasing time of preservation. Non significantly decrease of the semen quality with advance of preservation time indicates the suitability of the protocol used for freezing of indigenous ram semen in Bangladesh.DOI: http://dx.doi.org/10.3329/bjas.v44i1.23113            Bang. J. Anim. Sci. 2014. 44 (1): 10-15

62 EFFECTS OF VARYING GLUTATHIONE CONCENTRATIONS IN SEMEN EXTENDER ON THE QUALITY OF FROZEN–THAWED CANINE SPERM

2014 ◽  
Vol 26 (1) ◽  
pp. 145
Author(s):  
K. Ogata ◽  
B. Sarentonglaga ◽  
M. Yamaguchi ◽  
A. Sasaki ◽  
Y. Kato ◽  
...  
Keyword(s):  
Sperm Motility ◽  
High Speed ◽  
Pure Water ◽  
Egg Yolk ◽  
Motility Index ◽  
Term Survival ◽  
Genetic Traits ◽  
Frozen Semen ◽  
Semen Extender

Trans-cervical insemination (TCI) with cryopreserved semen offers a potentially effective approach for breeding canids with specific genetic traits, such as guide dogs for the blind. However, there are technical difficulties in canine sperm cryopreservation, such as the composition of semen extender. The aim of this study was to evaluate the effects of glutathione (GSH) as an antioxidant in the semen extender to improve the quality of frozen-thawed dog sperm. A Tris-egg yolk-citrate extender containing 15.7 mg mL–1 of TRIS, 8.8 mg mL–1 of citric acid, 14.1 mg mL–1 of lactose, 25.4 mg mL–1 of raffinose, 1% (vol/vol) antibiotics, and 20% (vol/vol) egg yolk in ultra-pure water was used as the base medium. Twelve ejaculates were collected from 7 dogs. Each ejaculate was divided into 2 to 5 aliquots and extended with base extender supplemented with 0, 2.5, 5, 7.5, and 10 mM GSH as first dilution. The extended semen was equilibrated for 3 h at 4°C. An equal volume of second extender was added to obtain a final concentration of 6.5% glycerol and sperm per milliliter. The sperm samples were loaded in straws and frozen at 6 cm above the surface of LN2 for 15 min in a styrene foam box and plunged into the LN2. The frozen semen was thawed for evaluation. The motility of sperm was estimated with a phase-contrast microscope and the motile patterns were classified into the following grades: progressively motile at a high speed (+++), progressively motile at a moderate and low speed (++), motile without progression (+), and immotile (–). Then, the sperm motility index (SMI) was determined from the following formula as described previously (Iritani et al., 1975), with some modifications: the percentage of (+++) sperm + the percentage of (++) sperm × 0.75 + the percentage of (+) sperm × 0.5. Sperm motility and the SMI were determined at 0, 1, 2, 3, 4, 12, and 24 h after thawing. Acrosome status was evaluated at 4 h after thawing. Lipid peroxidation (LP) levels at 0 and 12 h after thawing were used to examine the antioxidant ability of GSH. Trans-cervical insemination was carried out on 5 bitches to evaluate the fertility of GSH-treated sperm. The TCI were performed nonsurgically with a laparoscope and deposited 2 mL of semen through a catheter. Each bitch was inseminated 1 to 2 times during oestrus. Data were analysed using ANOVA with the Tukey-Kramer method. We found that the rate of (+++) sperm in the 5 mM GSH group was higher than that in the 0 mM group from 1 to 24 h after thawing (P < 0.05). The SMI was higher in the 5 and 7.5 mM GSH groups than in the 0 mM group (P < 0.05). There were no significant differences in the control and 2.5 and 10 mM GSH groups. Long-term survival was increased in the 5 mM GSH group. Acrosome integrity was higher in the GSH-treated group. The level of LP was lower in the GSH-treated groups at 0 h after thawing (P < 0.05). Trans-cervical insemination with the 5 mM GSH-treated semen resulted in the delivery of 5 pups from 2 bitches. These results indicate that the cryopreservation with 5 mM GSH can improve the motility, viability, and fertility of frozen-thawed canine sperm by its antioxidant effects on the sperm membrane.

scholarly journals The Influence of Breed and Type of Extender on the Quality of Bull Semen

2020 ◽  
Vol 21 (2) ◽  
pp. 64
Author(s):  
Iman Sukirman ◽  
Eros Sukmawati ◽  
Siti Darojah Rasad ◽  
Nurcholidah Solihati
Keyword(s):  
Plasma Membrane ◽  
Semen Quality ◽  
Block Design ◽  
Egg Yolk ◽  
Spermatozoa Motility ◽  
Bull Semen ◽  
Randomized Block Design ◽  
Frozen Semen ◽  
Two Factors

This study aimed to determine the influence of breed and type of extenders on frozen semen quality of cows at BIB Lembang. The experimental study was conducted in a Factorial Randomized Block Design (RBD) with two factors. The first factor was four cow breeds, i.e. Ongole Cross (PO), Brahman (BR), Simmental (SM) and Limousin (LM), and the second factor was two types of extender, i.e. Skim-Egg Yolk (SKT) and AndroMed® (AND), all repeated four times. The observed variables were percentage of spermatozoa motility and intact plasma membrane (IPM). All data obtained were analyzed using a general linear model (IBM SPSS ver. 23). The results demonstrated an interaction between breed and the type of diluent to motility. Breeds showed significantly different motility but non-significantly different intact plasma membrane (MPU) of semen. The type of diluent did not significantly affect motility and intact plasma membrane (MPU) of the frozen semen. The effect of the breed on BR motility was lower and significantly different from PO, LM and SM. The types of diluent did not significantly affect motility, MPU. The results showed that SKT was lower than AND, it was indicative effect of breed on intact plasma membrane (MPU) PO was lower than BR, LM and SM and the effect of the type of diluent on whole plasma membrane (MPU) AND is lower than SKT. It can be concluded that breed influences the motility of semen. The lowest motility reduction in frozen semen is Brahman cattle by using skim-egg yolk extender.

scholarly journals 98CONCEPTION RATES USING BRAHMAN BULL SEMEN FROZEN IN MILK BASED EXTENDER CONTAINING EGG YOLK OR SOYBEAN LIPIDS; A FIELD STUDY IN A TROPICAL ENVIRONMENT

2004 ◽  
Vol 16 (2) ◽  
pp. 170 ◽  
Author(s):  
R. Gonzales ◽  
M. Rosales ◽  
F. Perea ◽  
J. Velarde ◽  
E. Soto ◽  
...  
Keyword(s):  
Egg Yolk ◽  
Pregnancy Rates ◽  
Glycerol Concentration ◽  
Forest Environment ◽  
Chi Square ◽  
Bull Semen ◽  
Frozen Semen ◽  
The Mean ◽  
Semen Extender ◽  
Chi Square Analysis

The objective of this study was to examine the substitution of soybean-origin phospholipids for egg yolk in Brahman bull semen extender. Semen was frozen in 3 different low-fat milk (1%) based extenders containing 10mgmL−1 of fructose and supplemented with: 8% of whole egg yolk (Extender 1, control), 8% rectified egg yolk (egg yolk granules were removed by double centrifugation at 3000g for 1h at 5°C; Extender 2), and 7.3mgmL−1 of phospholipids of soybean-origin containing 10% of phosphatidyl choline (Extender 3). All 3 extenders were supplemented with 1000IU of penicillin, 1mgmL−1 streptomycin and 150μgmL−1 lincomycin. The semen was collected by means of artificial vagina from 3 Brahman bulls, and AI was performed during the dry season between December and April in a tropical forest environment. The mean temperature for the region was 26–30°C, with mean rainfall of 900–1500mm/year and the relative humidity of 60–70%. Ejaculates with at least 60% motility were diluted in 2 steps as follows: in step 1, each ejaculate was split into 3 even parts and diluted at 26°C with each of the extenders containing no glycerol, and in step 2, 14% of glycerol was added in 15-minute intervals to a final glycerol concentration of 7%. Semen was aspirated into 0.5mL plastic straws (20×106 sperm/per straw), frozen 7cm above liquid nitrogen (LN2) for 8min, and then plunged into LN2. Straws were thawed in a water bath at 37°C for 30s. Each experiment was replicated 3 times (different collection days). Sperm viability was tested within artificial insemination trials. Results are based on the pregnancy rates of crossbreed Brahman cows determined by palpation 45 Days after AI and by calving rates. Data were compared by chi-square analysis. In Experiment I, a total of 157 cows were inseminated with semen collected from 3 different bulls (A, B and C) and frozen in 3 different extenders (1, 2 and 3; 3×3 factorial design). Bull A, Extender 1, 2 and 3 (n=19, 20 and 22); Bull B, Extender 1, 2 and 3 (n=20, 20 and 20) and Bull C, Extender 1, 2 and 3 (n=22, 15 and 24), respectively. Although semen from all 3 bulls frozen in Extenders 2 and 3 fostered numerically higher pregnancy rates (from 30% for Bull B and Extender 2 to 50% for Bull C and Extender 3) than in Extender 1 (from 23.5% for Bull C to 40% for Bull B), there were no differences (P&lt;0.05) between bulls with any of 3 extenders on the pregnancy rates. In Experiment II, a total of 117 cows were inseminated with semen collected from Bull B and frozen in Extender: 1 (n=37), 2 (n=48) and 3 (n=39). There were significantly higher (P&lt;0.05) calving rates for cows inseminated with semen frozen in Extender 2 and 3 (41.6% and 46.1%, respectively) than in Extender 1 (24.3%). It can be concluded that rectified egg yolk may improve viability of frozen semen, and that phospholipids of soybean origin can be successfully substituted for egg yolk in Brahman bull milk based semen extender. Supported by Bioniche Inc, Belleville, Ontario, Canada.

scholarly journals Quality of Sexed Sperm of Bali Bull in Regional Artificial Insemination Center of Riau Province

2019 ◽  
Vol 43 (2) ◽  
Author(s):  
Yendraliza Yendraliza ◽  
Anwar Efendi Harap ◽  
July Handoko ◽  
Muhammad Rodiallah
Keyword(s):  
Plasma Membrane ◽  
Pregnancy Rate ◽  
Artificial Insemination ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Female Offspring ◽  
Plasma Membrane Integrity ◽  
Frozen Semen ◽  
Calving Rate

This study aimed to evaluate the quality of frozen semen of Bali bull resulted from sexing procedure on calf or offspring production with desired sex. The tested sperm of Bali bull were collected from Bali bull raised at Regional Artificial Insemination Center of Riau Province (BIBD Riau). The study was carried out in 2 stages. The first stage was X and Y chromosome separation by albumin method. The extender used in the sexing procedure is trice citrate fructose and egg yolk. The second stage was mainly testing the sexed sperm collected in 60 Bali cow in Langkat Village, Bengkalis Regency. To determine the quality of post thawing frozen semen collected from the sexing procedure, the study evaluated motility, viability, mortality, abnormality and plasma membrane integrity of the spermatozoa. The pregnancy rate, calving rate, and birth accuracy of inseminated sexed sperm to offspring’ sex were also evaluated. The evaluation resulted in motility (66.3-75.3%), viability (70-78.5%), plasma membrane integrity (60-65.8%), abnormality (6.05-8.05%), mortality (20.05-30.05%), and pregnancy rate (83.33-90%). The calving rate on this study was 100% with the birth accuracy of 81.8% for male offspring and 40% for female offspring. As conclusion, the sexed sperm evaluated on this study have fairly good fertility.

scholarly journals Kualitas Sperma Beku Sapi Bali dalam Pengencer Air Kelapa Modifikasi dengan Berbagai Aras Dimethyl Sulfoxide (FROZEN SPERM QUALITY OF BALI BULLS IN MODIFIED COCONUT WATER EXTENDER WITH DIFFERENT DIMETHYL SULFOXIDE CONCENTRATION)

2019 ◽  
Vol 20 (1) ◽  
pp. 93
Author(s):  
Thomas Mata Hine ◽  
Kirenius Uly ◽  
Wilmientje Marlene Nalley ◽  
Heri Armadianto
Keyword(s):  
Dimethyl Sulfoxide ◽  
Liquid Nitrogen ◽  
Sperm Quality ◽  
Egg Yolk ◽  
Optimal Concentration ◽  
Coconut Water ◽  
Low Molecular Weight ◽  
Frozen Semen ◽  
Artificial Vagina

Dimethyl Sulfoxide (DMSO) is one type of cryoprotectant which has a low molecular weight so that it is easier to enter cells when cryopreservation. The purpose of this study was to explore the optimal concentration of DMSO in modified coconut water (mCW) extender that were able to maintain frozen sperm quality of bali bulls. Semen was collected from two four-year old bali bulls by artificial vagina. Good quality semen diluted with mCW (young coconut water + 20% egg yolk + 7.5 % moringa leaf extract) and supplemented by 3, 5, or 7% DMSO. Semen was filled into 0.25 ml ministraw, and was incubated in a refrigerator at 5°C for four hours, frozen on the surface of liquid nitrogen for 10 minutes and then dipped into liquid nitrogen. The quality of post thawing sperm was measured 24 hours later by placing the ministraw of frozen semen into water at 37oC for 30 seconds. Data were analyzed by analysis of variance and continued with Duncan test. Postthawing observations showed that bali bulls sperm cryopreserved at 3% DMSO yielded higher motility and viability (p<0.05) i.e. 36 and 44.15%, than DMSO 5% i.e. 18 and 23.65%, and DMSO 7% i.e. 7 and 12.62%. The recovery rate of sperm cryopreserved at 3% DMSO was also higher (p<0.05) than DMSO 5 and 7%, successively 45.65, 23.06, and 8.86%. The results of this study concluded that the optimal concentration of DMSO in mCW diluent to maintain frozen sperm quality of bali bulls was 3%. 

scholarly journals Supplementation of Pandanus conoideus Oil in Cryopreservation Diluents for Maintaining the Semen Quality of Ongole Grade Bull

2021 ◽  
Vol 44 (2) ◽  
pp. 146-151
Author(s):  
Nurcholis Nurcholis ◽  
A. Furqon ◽  
R. I. Arifiantini ◽  
S. M. Salamony
Keyword(s):  
Sperm Motility ◽  
Recovery Rate ◽  
Semen Quality ◽  
Egg Yolk ◽  
Sperm Viability ◽  
Nitrogen Vapor ◽  
Frozen Semen ◽  
Fruit Oil ◽  
Range Test

Antioxidants such as tocopherol, ß-carotene, and polyunsaturated fatty acids (PUFA) from red fruit oil of Papua may be used to protect frozen semen. The study is aimed to test the effect of red fruit oil supplementation on motility, viability, and recovery rate of frozen sperm of Ongole-grade bulls. Semen was collected twice a month from eight 4-5-year-old male Ongole grade using an artificial vagina, followed by macro- and microscopical evaluations. Collected semen was divided into four tubes and diluted with tris egg yolk diluents (TEY) as a control, TEY supplemented with 0.5% red fruit oil (RFO) (TEY RFO0.5), TEY supplemented with 1% RFO (TEY RFO1.0), and TEY supplemented with 1.5% RFO (TEY RFO1.5). The diluted semen was then packed into the straw and equilibrated for 2, 4, and 6 h prior to frozen on liquid nitrogen vapor for 10 minutes. The observed variables in this study were sperm motility, sperm viability, and morphology after equilibration, after thawing, and recovery rate. The experimental design is a completely randomized factorial design. The data were analyzed using ANOVA and were further tested using Duncan multiple range test. The results showed that the sperm motility of fresh semen was 81.10±1.42%. The percentage of sperm motility in TEY RFO1.5 treatment at 6 h equilibration was 60.00±1.06%, significantly higher compared to TEY RFO1.0 and TEY RFO0.5. The percentage of post-thawing sperm motility in TEY RFO1.5 treatment was 62.40±1.09%. The best post-thawing sperm viability in TEY RFO1.5 was 80.70±1.20%, significantly increase from the treatment of TEY RFO1.0 and TEY RFO0.5. The recovery rate (RR) for TEY RFO1.5 treatment had the best percentage at 76.94%. In conclusion, RFO supplementation in semen diluents for 2 h of equilibration resulted in the best motility and viability at 0 h of post thawing observation.

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