scholarly journals Using Gum Arabic in Place of Egg Yolk as Cryoprotectant for Cryopreservation of the Buck Semen

2021 ◽  
Author(s):  
Mohamed Ali
Keyword(s):  
Pregnancy Rate ◽  
Gum Arabic ◽  
Egg Yolk ◽  
Fertility Rate ◽  
Control Group ◽  
Motile Sperm ◽  
Frozen Semen ◽  
Different Levels ◽  
Motility Rate

Background: Egg yolk (EY) is well known to be toxic for buck spermatozoa, which creates restrictions of its use in cryopreservation. Therefore, this study is to compare the effect of different levels of gum Arabic (GA) in an extender on quality and fertility of cryopreserved buck sperm. Methods: Each ejaculate of six bucks was frozen in Tris with one of concentrations of GA which contained 3, 6, 9 and 12 gm/ 100 ml in place of the EY. Control was Tris extender containing 2.5% of EY. Result: A percentages of total motile sperm (54.92%; P less than 0.05) and progressively motile sperm (26.22%; P less than 0.05) of semen was frozen in Tris containing 9% of GA. Similar to control group, the pregnancy rate of does inseminated with extender containing 9% (50.0%) were significantly higher than those of does inseminated with extender containing 6% (8.33%), 3% (0.0%) and 12% (0.0%). Semen evaluation and fertility rate were similar when replacing the EY with GA in the Tris cryodiluent, after cryopreservation of buck semen. The present study shows that high motility rate of frozen semen and acceptable pregnancy rate can be obtained when using GA in place of EY for cryopreserving the buck sperm.

14 MOTILITY AND FERTILITY OF BULL SPERMATOZOA FROZEN IN EGG YOLK EXTENDER SUPPLEMENTED WITH LACTOFERRIN

2007 ◽  
Vol 19 (1) ◽  
pp. 125 ◽  
Author(s):  
J. Kobayashi ◽  
Y. Suda ◽  
N. Takada ◽  
K. Komine ◽  
T. Kuroishi ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Gradient Centrifugation ◽  
Bovine Milk ◽  
Egg Yolk ◽  
Field Trials ◽  
Fertility Rate ◽  
Control Group ◽  
Motile Sperm ◽  
Percoll Gradient ◽  
Frozen Semen

Lactoferrin (Lf) is secreted by the prostate, seminal vesicles, and epididymis, and appears to represent a major component of the sperm-coating antigens that protect sperm. The aim of this study was to determine the effects of Lf used in semen extender on motility and fertility of bovine frozen semen. Semen samples were collected from three fertile Japanese Black sires. The semen collected was cooled and extended with egg yolk–Tris extender supplemented with glycerol and Lf (from bovine milk) to give final concentrations of 7% and 0, 10, 100, 500, 1000, or 2000 �g mL-1, respectively. After being extended, the semen was packaged in 0.5-mL straws, frozen, and transferred into liquid nitrogen. At least one week later, the frozen semen was thawed and washed mTALP containing caffeine. Motile sperm were separated by discontinuous 2-step (40 and 80%) Percoll gradient centrifugation. Sperm motility was calculated as the percentage of motile sperm separated by Percoll gradients relative to pre-separated sperm. Sperm motility character (motile sperm, progressive sperm) was analyzed by a CASA system (Hamilton Thorne Biosciences, Beverly, MA, USA). In the following experiment, we examined the effect of Lf added in the extender on improvement of semen fertility. The frozen semen from one sire extended with or without 500 �g mL-1 of Lf was prepared (99 straws with Lf, 99 straws without), distributed to three inseminators, and used to inseminate to 198 cows. The sperm motility and motility character were analyzed by ANOVA and Fisher's exact test. The field trials were evaluated by chi-square test. After selection of motile sperm with the Percoll gradient, examination of sperm motility revealed that addition of Lf in the extender increased sperm motility significantly (P < 0.05) compared to that in the control without Lf: 51.2–53.8% in semen with added Lf (100–2000 �g mL-1) vs. 39.6% in the control. CASA analysis showed that motile sperm and progressive sperm were improved significantly by the addition of Lf; the peak level was 500 �g mL-1 of Lf in the extender. The percentage of motile sperm in semen with added Lf was higher significantly (P < 0.01) than that of the control: with Lf of 100 and 500 �g mL-1, 50.8 and 56.0%, respectively, vs. 39.7% without Lf. The percentage of progressive sperm was also significantly higher (P < 0.05) than that of the control: with Lf of 100 and 500 �g mL-1, 18.8 and 19.0%, respectively, vs. 13.5% without Lf. In the next experiment, we performed field trials to assess the availability of Lf for artificial insemination. In total, the group with Lf showed some but not a significant increase in fertility rate (delivery cow/inseminated cow) compared with the control group without Lf: 56.6% vs. 52.5%, respectively. In conclusion, these results suggest that addition of Lf in sperm extender may be useful for improvement of sperm motility and motility character after freezing and thawing. In the present study, we used fertile bulls, but the concentration of intact Lf in the semen was unknown, which might be the reason why the effect of Lf on the fertility rate was not observed to be significant. In future experiments it would be valuable to demonstrate the effect of Lf in subfertile semen.

scholarly journals Effect of Gum Arabic on Stallion Sperm Survival During Cold Storage and Post Freezing

2018 ◽  
Vol 41 (1) ◽  
pp. 21-31
Author(s):  
Mohamed Ali ◽  
Musa M. Musa ◽  
Sulaiman Alfadul ◽  
K. Al-Sobayel
Keyword(s):  
Gum Arabic ◽  
Egg Yolk ◽  
High Viscosity ◽  
Motile Sperm ◽  
Sperm Velocity ◽  
Low Viscosity ◽  
Straight Line ◽  
Frozen Semen ◽  
Sperm Survival ◽  
Semen Extender

Abstract This study is aimed at investigating effects of supplementation of stallion’ semen extender with various concentrations of Gum Arabic (GA) versus egg yolk (EY) on viscosity, sperm motility and survival during cooling and freezing. Physical sperm characteristics; i.e. curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), linearity (LIN) and straightness index (STR) were evaluated. Based on the sperm velocity (velocity of the average path), individual spermatozoons were classified into two major groups; i.e., progressively motile (>45 μm/sec) and immotile (0-45 μm/sec) spermatozoa. Addition of 3, 9 or 15% of GA to HF-20 extender resulted in linear decreases in VCL, VSL and VAP and a decrease in the percentage of progressively motile spermatozoa. Dilution of horse semen samples with high viscosityextenders (i.e., high percentage of GA) decreased the VCL, VSL and VAP in fresh and chilled semen. Freezing semen in high viscosity-extenders reduced percentage of progressively motile spermatozoa compared with those of low viscosity-extenders. In refrigerated and frozen semen samples, the extender containing 15% GA had detrimental effects on the percentage of progressively motile sperm cells and velocity of progressive motile sperm. Moreover, cooling sperm in extenders containing 9 or 15% of GA for 72 hours resulted in complete motility cessation. In conclusion, GA could replace EY in stallion semen extenders at a level of 3% to maintain the physical and biological characteristics of cold and frozen semen.

scholarly journals Effect of green tea extract in extender of Simmental bull semen on pregnancy rate of recipients

2021 ◽  
Vol 34 (2) ◽  
pp. 198-204 ◽  
Author(s):  
Suherni Susilowati ◽  
Trilas Sardjito ◽  
Imam Mustofa ◽  
Oky Setio Widodo ◽  
Rochmah Kurnijasanti
Keyword(s):  
Pregnancy Rate ◽  
Dna Fragmentation ◽  
Green Tea ◽  
Prostaglandin F2α ◽  
Green Tea Extract ◽  
Control Group ◽  
Bull Semen ◽  
Frozen Semen ◽  
Tea Extract

Objective: The aim of this study was to ascertain the effects of adding green tea extract (GTE) to skim milk-egg yolk (SM-EY) extender on both the quality of post-thawed bull semen and the pregnancy rates of the recipient cows.Methods: Twelve ejaculates from four Simmental bulls, aged 3 to 5 years and weighing 900 to 950 kg, were diluted SM-EY extender, added with 0, 0.05, 0.1, and 0.15 mg GTE/100 mL extender and then frozen. After four weeks storage in liquid nitrogen, the sperm were thawed and evaluated for viability, motility, intact plasma membrane (IPM), and DNA fragmentation. Meanwhile, the estrus cycles of 48 recipient cows were synchronized by intramuscular administration of a single injection of 5 mg prostaglandin F2α. Estrus cows were divided into four equal groups and inseminated artificially 18 to 20 h after the onset of estrus by using semen from each extender group. Pregnancy was diagnosed by measuring serum progesterone levels at 21 days, followed by transrectal palpation 90 days after insemination.Results: The findings revealed that adding 0.1 mg of GTE/100 mL extender produced the highest percentages of sperm viability (70.67%±1.75%), motility (69.17%±1.47%), and IPM (69.23%±1.21%) and the lowest percentage of DNA fragmentation (3.00%±0.50%). The pregnancy diagnosis revealed that all cows (36/36) inseminated using frozen semen in GTE addition extender were pregnant (pregnancy rate 100%), whereas the pregnancy rate of the control group was 83.33% (10/12).Conclusion: It may be concluded that 0.1 mg GTE/100 mL extender yields the best quality of spermatozoa and that all variants doses of GTE in extender produce a higher pregnancy rate among recipient cows.

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.

34 The effect of dilution method of beagle dog semen on the survival rate of cryopreserved spermatozoa after thawing

2019 ◽  
Vol 31 (1) ◽  
pp. 143
Author(s):  
S. W. Kim ◽  
C.-L. Kim ◽  
I. S. Jeon ◽  
Y. G. Ko ◽  
I.-S. Hwang
Keyword(s):  
Drug Testing ◽  
Semen Analysis ◽  
Egg Yolk ◽  
Control Group ◽  
Dilution Method ◽  
Computer Assisted ◽  
Motile Sperm ◽  
Optimal Method ◽  
Beagle Dog ◽  
Progressive Motility

The successful cryopreservation of spermatozoa of the beagle dog for AI is essential for the establishment of the genetic banks of drug detection dogs. The beagle dog is widely used for drug testing and chosen for breeding by breeders. However, the use of cryopreserved beagle semen is limited by the lower number of offspring of dog species. In this study, 3 highly trained beagle dogs were chosen and their semen was cryopreserved for the next generation. The effects of dilution methods of beagle semen were tested using a direct dilution method at RT and a 2-step dilution method at 5°C. As a control group, the effects of a direct dilution method of semen on the percentage of motile sperm and progressive motility were analysed by computer-assisted semen analysis system (SAIS, Korea), and abnormality of spermatozoa was examined by Diff Quik staining. A total of 9 samples from 3 dogs were extended in 4% glycerol containing Tris-egg yolk diluents at approximately 22 to 25°C. The diluted semen was cooled to 5°C within 2h. The packed 0.5-mL straws were placed 5cm above the surface of LN for 10min and then plunged in. A 2-step dilution method was conducted using the same procedures of freezing, but the first dilution was done with glycerol-free diluent. After cooling to 5°C within 2h, the second diluent with 8% glycerol was added to the same volume of diluted semen at 5°C and stabilised for 1h. After thawing for 45s at 37°C, the semen from the 2-step dilution method showed the higher percentage of motile sperm (65.4±6% v. 45.3±8%; P&lt;0.05) and progressive motility (41.6±5.3% v. 32.3±3.7%; P&lt;0.05). However, the abnormalities between groups showed no differences. The results suggest that the optimal method for freezing beagle dog spermatozoa is a 2-step dilution process that consists of the first dilution at RT and the second dilution with glycerol at 5°C into diluted semen.

50 Effects of Cathepsin B Inhibitor E64 on the Survival Rate of Cryopreserved Semen from Korean Brindled Bulls

2018 ◽  
Vol 30 (1) ◽  
pp. 164
Author(s):  
S. W. Kim ◽  
M.-S. Kim ◽  
C.-L. Kim ◽  
I. S. Jeon
Keyword(s):  
Cathepsin B ◽  
Room Temperature ◽  
Egg Yolk ◽  
Coat Colour ◽  
Control Group ◽  
Vitro Fertilization ◽  
Frozen Semen ◽  
The Difference ◽  
The Individual

Korean brindled cattle have distinctive coat colour and are regarded as rare cattle in the Korean peninsula. To preserve the line as a genetic resource for diversity in cattle, semen cryopreservation has been used for selection of individuals for breeding between local AI centers. Nevertheless, the survival and viability of frozen semen from Korean brindled bulls is not uniform due to the difference between the individual bulls or experimental techniques. In this study, E64, a cathepsin B inhibitor, was used at final concentrations of 0.05, 0.5, and 1 µM to test the viability of frozen semen. To prepare frozen semen, a triladyl-egg yolk diluent with 6.4% glycerol was used in a two-step freezing method with 4 different bulls with 3 repeats. A total of 12 ejaculates was diluted to a concentration of 50 to 100 × 106 mL−1 at room temperature, and slowly cooled from room temperature to 5°C in 2 to 3 h. The cooled semen was diluted 1:1 with a secondary diluent containing E64 to prepare the experimental group. After loading, 0.5-mL straws were immersed into liquid nitrogen after 10 min exposure at 5 cm above the nitrogen using a styrofoam box. The viability of spermatozoa after thawing at 37°C for 40 s was analysed by Student’s t-test. The rate of surviving sperm in the 1 µM E64 group (82 ± 4.3%) was significantly higher than that of the control group (71.2 ± 2.0%; P < 0.05). However, the 0.05 and 0.5 µM E64 treatment groups lead to similar rates (77.5 ± 2.0% and 75.0 ± 5.0%, respectively; P > 0.05). Based on these results, it is expected that E64 could be used for the improvement of productivity of frozen semen; further results on in vitro fertilization and development are ongoing.

scholarly journals Introducing Soybean Oil in Semen Extenders of Awassi Rams

2018 ◽  
Vol 12 (1) ◽  
pp. 86-91
Author(s):  
Nameer Al-biaty ◽  
Safaa Abed ◽  
Hazim J. Alobaidi ◽  
Safaa Almaliki ◽  
Abbas F Kareem ◽  
...  
Keyword(s):  
Soybean Oil ◽  
Semen Quality ◽  
Storage Period ◽  
Egg Yolk ◽  
Control Group ◽  
Significant Deterioration ◽  
Sperm Abnormality ◽  
And Storage ◽  
Pooled Samples ◽  
Different Levels

The aims of the study was to investigate the possibility of using an plant lecithin; commercial soybean oil (SO) directly in the components of semen extenders of Awassi rams, storage of semen in chilled technique and the effects of dilution, cooling and storage periods on semen quality. Semen was collected weekly from four Awassi rams by electro- ejaculator. Pooled samples were divided to six equal aliquots and diluted by Citrate egg yolk extender at 37˚C. Treatments were designed on the base of control extender containing 25% egg yolk and four treatments containing different addition of SO (12.5, 25, 37.5, and 50%) and combination treatment of 12.5% egg yolk + 12.5% SO. Treatment tubes were cooled to 5˚C and stored for three days. Semen was evaluated as raw, diluted, cooled and after storage in refrigerator (5˚C) for 1, 2 and 3 consecutive days. Results showed that there were no significant differences among all treatments in progressive motility, dead sperm %, abnormal sperm % and acrosome defect %, while pH were found to be significantly declined (p≤ 0.05) in control group. Significant effects of dilution, cooling, and storage period have been demonstrated with steadily significant deterioration (p≤ 0.05) for all studied characteristics when motility and pH declined while sperm abnormality, dead, and acrosome defect percentages increased. The results clearly indicated successful of using different levels of SO (plant source) as lecithin source instead of egg yolk (animal source of lecithin) without any impacts on the biological characteristics of Awassi ram semen and the process of dilution, cooling and storage periods have deterioration effects on semen quality.

scholarly journals 10CASA EVALUATION OF SEXED AND NON-SEXED FROZEN BULL SEMEN

2004 ◽  
Vol 16 (2) ◽  
pp. 127 ◽  
Author(s):  
G. Brogliatti ◽  
G. Barreiro ◽  
G. Larraburu ◽  
A. Laborde
Keyword(s):  
Sperm Motility ◽  
High Speed ◽  
Sperm Quality ◽  
Sperm Concentration ◽  
Glass Tube ◽  
Egg Yolk ◽  
Motile Sperm ◽  
Bull Semen ◽  
Frozen Semen ◽  
Sexed Semen

Flow citometry cell sorting has been proven successfully to separate X and Y sperm; however, the technology is still too stressfull for the viability of the sorted semen. The objective of this study was to evaluate nonsexed and sexed frozen sperm motility characteristics using a CASA technology. Ejaculates from 4 different bulls (3 Holstein and 1 Angus) were collected, and processed as split non-sexed and sexed semen samples using Tris egg yolk extenders. X and Y sperm were separated using a high-speed sorter (SX Moflo). Cryopreservation was done at the same time under appropiate conditions using a programmed cryochamber. Thawing procedure was done at 37°C for 30s and a sample of each straw was placed in the evaluation chamber. The experiment was repeated twice and two chambers with 30 observations each were analyzed each time. Mean and standard deviation of each characteristic were calculated, compared and analyzed statistically. The sperm concentration was determined by means of a burker counting chamber. Sperm quality was determined at 0h after thawing, and later at 1h, 2h and 3h after incubation in a glass tube at 30°C. The following sperm motility parameters were determined with the Hamilton Thorne (HTM-ceros 12.1) on at least 1000 spermatozoa: velocity average path (VAP), velocity straight line (VSL), amplitude lateral head (ALH), beat cross frequency (BCF), straightness (STR), linearity (LIN), and percentage of progressively motile spermatozoa (PMS). Linearity of nonsexed spermatozoa was 53±3.5, 47±0.8, 43±7.8 and 42±4.5 for the 0h and the 3 test incubation times and 49.5±3.7, 51.2±3.7, 43.3±7.8 and 44.5±7.6, respectively, for sexed semen. There were no significant differences (P&gt;0.05) in the progressive velocity, track speed and linearity between sexed and nonsexed semen. The percentage of static cells was 33%, 30%, 47% and 50% for the 0h and the 3 test incubation periods; however, the percentage of static cells for the sexed semen was 53%, 71%, 77% and 82%, respectively. Results from the analysis indicate a significant increase (P&lt;0.01) in the number and the percentage of static cells with time. The lateral amplitude of sperm motility for nonsexed semen was 5.9±0.5, 6.8±0.8, 6.0±0.4 and 5.1±0.7, and for sexed semen 6.6±0.7, 6.8±0.4, 6.4±0.4 and 5.5±1.7, respectively. The percentage of progressively motile sperm was significantly different at 0 time 49.7±4.9 and 23.1±4.9 for nonsexed and sexed semen respectively. After 3 hours of incubation the percentage of progressively motile sperm was 38.7±10.2 and 3.7±3.2 for nonsexed and sexed semen, respectively. In conclusion, sexed frozen semen seems to have characteristics similar to those of normal nonsexed semen. However, a significant decrease in the percentage of progressively motile cells could affect pregnancy rates. More research needs to be done to detect differences between bulls and cryoprotectans.Research supported by Centro Genetico Bovino de EOLIA sa Argentina.

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.

Sign in / Sign up

Export Citation Format

Share Document