The effect of soybean extract supplementation to tris aminomethane–egg yolk extender on the post-thawing quality of Simmental semen

This research aims to evaluate the effect of soybean extract (Glycine max (L.) Merr.) supplementation on tris aminomethane egg yolk extender to the post thawing quality of Simmental bull frozen semen. The fresh semen was collected from 3 Simmental bulls aged at 3 to 3.5 years for twice a week by using artificial vagina. The semen was then selected for sperm motility and abnormality, and the semen that had >70% motility and <15% abnormality was used for the research. The research was conducted in a completely randomized with 4 treatments and 30 replications. The research treatments include 0% (T0), 7.5% (T1), 10% (T2), and 12.5% (T3) soybean extract supplementation on tris aminomethane egg yolk extender. The observed variables include spermatozoa motility, viability, abnormality, and plasm membrane integrity. The collected data were analysed with ANOVA and followed with Duncan’s Multiple Range Test to determine significant differences. The results showed that soybean extract supplementation gave significant effect (P<0.05) to the post thawed quality of Simmental semen. The research concludes that 10% (v/v) soybean supplementation on tris aminomethane egg-yolk extender was effective to maintain sperm motility, viability, and plasma membrane integrity of post thawed Simmental semen.

Download Full-text

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

Tropical Animal Science Journal ◽

10.5398/tasj.2021.44.2.146 ◽

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.

Download Full-text

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

Bangladesh Journal of Animal Science ◽

10.3329/bjas.v44i1.23113 ◽

2015 ◽

Vol 44 (1) ◽

pp. 10-15 ◽

Cited By ~ 1

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

Download Full-text

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

Reproduction Fertility and Development ◽

10.1071/rdv26n1ab62 ◽

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.

Download Full-text

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

ANIMAL PRODUCTION ◽

10.20884/1.jap.2019.21.2.641 ◽

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.

Download Full-text

Effects of apple and orange juices on quality of refrigerated goat semen

Journal of Agricultural Sciences Belgrade ◽

10.2298/jas1801053a ◽

2018 ◽

Vol 63 (1) ◽

pp. 53-65

Author(s):

Ezekiel Adekunle ◽

James Daramola ◽

Olusiji Sowande ◽

John Abiona ◽

Monsuru Abioja

Keyword(s):

Sperm Motility ◽

Orange Juice ◽

Apple Juice ◽

Membrane Integrity ◽

Egg Yolk ◽

Fruit Juices ◽

Sperm Viability ◽

In Vitro Storage

This study investigated the effects of apple and orange juices on quality of refrigerated spermatozoa of goat bucks. Semen samples from WAD goat bucks were diluted with Tris-egg yolk extenders each supplemented with apple and orange juices at 0, 2.5, 5, 7.5 and 10/100 ml of diluents. The diluted semen samples were assessed for sperm viability and malondialdehyde (MDA) concentration after in vitro storage for 240 hours at 5oC. The ability to maintain sperm motility was higher in the extenders with 7.5% orange juice followed by 10% apple juice compared to other treatments (P<0.05). The extenders supplemented with 2.5%, 5% and 7.5% apple juice, and 5% orange juice had higher intact acrosome compared to other treatments and the control (P<0.05). The 10% orange juice had higher percentage membrane integrity compared to other treatments. Consistent and reduced (P<0.05) MDA levels were observed in the extenders supplemented with fruit juices and lower MDA was observed in the extenders supplemented with 10% apple juice compared to other treatments and the control (P<0.05). The findings reveal that additions of the fruit juices to semen extenders to maintain the viability of refrigerated spermatozoa were best at concentrations of 10 ml/100 ml of apple juice and 7.5 ml/100 ml of orange juice.

Download Full-text

Individual variation in fresh and frozen semen of Bali bulls (Bos sondaicus)

Veterinary World ◽

10.14202/vetworld.2020.840-846 ◽

2020 ◽

Vol 13 (5) ◽

pp. 840-846

Author(s):

R. Indriastuti ◽

M. F. Ulum ◽

R. I. Arifiantini ◽

B. Purwantara

Keyword(s):

Dna Fragmentation ◽

Sperm Motility ◽

Individual Variation ◽

Sperm Concentration ◽

Individual Factors ◽

Abnormal Morphology ◽

Frozen Semen ◽

Semen Volume ◽

Fresh Semen

Aim: This study aimed to analyze the individual factors influencing the sperm quality of Bali bulls at Baturiti Artificial Insemination (AI) center. Materials and Methods: Semen that was ejaculated from nine Bali bulls was collected using artificial vaginas (n=5/bull). Semen ejaculates were evaluated immediately after collection to measure the quality of the fresh semen, including semen volume, sperm concentration, progressive motility, membrane integrity (MI), and abnormal morphology. Frozen semen was evaluated for progressive sperm motility, concentration, viability, MI, abnormal morphology, and deoxyribonucleic acid (DNA) fragmentation. Other secondary data, focusing on semen quantity (semen volume and sperm concentration), were also collected from frozen the semen production data of the Baturiti AI center from 2017 to 2019. Data were analyzed statistically using a completely randomized design, and one-way analysis of variance was applied to find differences among individual bulls. Results: Significant differences (p<0.05) were found among the bulls in semen volume, sperm motility, concentration, and MI of the fresh semen. Significant differences (p<0.05) were also found among the bulls in sperm motility, viability, MI, abnormal morphology, and DNA fragmentation of the frozen semen. Conclusion: Individual variation in all the tested sperm parameters of the fresh semen of Bali bulls, except sperm viability and abnormalities, was noted. Similarly, individual variation in all the tested sperm parameters in frozen semen, except sperm concentration, was noted. Therefore, individual factors can be used for selecting a superior bull in Bali cattle.

Download Full-text

Reduction Spermatozoa Motility and Viability on Various Local Chickens During Storage at 5℃

ANIMAL PRODUCTION ◽

10.20884/1.jap.2020.22.3.64 ◽

2021 ◽

Vol 22 (3) ◽

pp. 158-162

Author(s):

Nu'man Hidayat ◽

Chomsiatun Nurul Hidayah ◽

Aras Prasetiyo Nugroho

Keyword(s):

Vitamin E ◽

Sodium Dodecyl Sulfate ◽

Sperm Motility ◽

Sodium Dodecyl ◽

Egg Yolk ◽

Sperm Viability ◽

Spermatozoa Motility ◽

Abdominal Massage ◽

Range Test ◽

Local Chickens

The research compared the changes motility and viability of sperm from various local chickens during storage at 5℃ for 72 hours. Semen was collected every three days using the dorsal-abdominal massage from twelve chickens consisting of Kedu, Sentul and Pelung chicken. semen was diluted in extender contains 90% lactate ringer and 10% egg yolk extender with 0.025% sodium dodecyl sulfate and 2% vitamin E (LREYSE). Sperm motility and viability was observed every 12 hours and the measurements were made up to 72 hours of storage. Complete random design repeated measurement with 4 replications was used in this study. One-way analysis of variance was used to analyze the data and followed by Duncan’s Multiple Range Test. The results showed sperm motility declined and sperm viability reduced during storage for all breeds. The significant declined of motility between breeds were only observed at 60 and 72 hours and the reduced viability between breeds was observed at 72 hours of storage. The declined sperm motility for Kedu chicken (31.59±3.26% and 75.36±1.25%) and Pelung chicken (36.11±4.05% and 75.83±5.34%) were significantly lower (p<0.05) than Sentul Chicken (50.39±2.60% and 95.00±5.00%) at 60 and 72 hours of storage respectively, while the decline sperm viability of Kedu chicken (57.59±3.64%) and Pelung chicken (54.39±5.73%) was significantly lower (p<0.05) than Sentul Chicken (90.30±9.70%) after 72 hours storage. It can be concluded that the reduction sperm motility and viability of Kedu and Pelung chicken is lower than Sentul chicken which are stored at 5℃ for 72 hours.

Download Full-text

Freezability of Andalusian donkey (Equus asinus) spermatozoa: effect of extenders and permeating cryoprotectants

Reproduction Fertility and Development ◽

10.1071/rd14449 ◽

2016 ◽

Vol 28 (12) ◽

pp. 1990 ◽

Cited By ~ 15

Author(s):

D. Acha ◽

M. Hidalgo ◽

I. Ortiz ◽

M. J. Gálvez ◽

J. J. Carrasco ◽

...

Keyword(s):

Sperm Motility ◽

Sperm Quality ◽

Membrane Integrity ◽

Egg Yolk ◽

Equus Asinus ◽

Sperm Membrane ◽

Freeze Test ◽

Pooled Samples

The aim of this study was to compare the effect of two semen extenders and four permeating cryoprotectants on post-thaw sperm quality of Andalusian donkeys. First, 32 ejaculates were pooled, split and frozen in either Gent B or INRA 96 with egg yolk and glycerol. Second, 12 pooled semen samples were simultaneously frozen in Gent B (glycerol) or Gent A containing ethylene glycol (EG; 1 or 1.5%) or dimethyl sulfoxide (DMSO; 1.5 or 2%). Finally, nine pooled samples were simultaneously cryopreserved in Gent A containing 1% EG (as control), dimethylformamide (DMFA; 1 or 2.5%) or a combination of 1% EG and 1.5% DMFA. Gent B yielded a higher (P < 0.01) post-thaw sperm motility than modified INRA96. EG 1% increased the sperm membrane integrity (P < 0.001), whereas DMSO affected sperm motility and membrane integrity (P < 0.001). DMFA 2.5% yielded higher (P < 0.001) values for sperm motility and membrane integrity. We concluded that Gent B improves in vitro post-thaw sperm quality of donkey spermatozoa, but the replacement of glycerol with 1% EG or 2.5% DMFA increased sperm protection against cryodamage. The use of DMSO for freezing donkey semen was unsuccessful and a toxic effect is suspected. These extenders should be included in the pre-freeze test for each donkey.

Download Full-text

Comparative Evaluation of Motility and Kinematics of Fresh Versus Frozen-Thawed Spermatozoa of Cattle and Buffalo Bull by CASA

Indian Journal of Animal Research ◽

10.18805/ijar.b-3881 ◽

2019 ◽

Author(s):

P K Pathak ◽

A J Dhami ◽

D V Chaudhari ◽

K K Hadiya

Keyword(s):

Sperm Motility ◽

Motile Sperm ◽

Mean Values ◽

Head Displacement ◽

Straight Line ◽

Frozen Semen ◽

Bovine Spermatozoa ◽

Motion Characteristics ◽

Fresh Semen

A study was undertaken on semen of three mature bulls each of Gir, Surti and Murrah breed to evaluate the comparative motion characteristics and kinematics of their fresh and frozen-thawed spermatozoa by Biovis CASA. The ejaculates (n= 24/breed) having >75% initial motility were diluted @ 80 million sperm/ml using TFYG extender and were assessed. Amongst motility traits, the total motile, rapid progressive motile and slow progressive motile spermatozoa percentage decreased significantly by 23.08 - 30.09, 43.57 - 55.18, 9.12 - 22.75 %, respectively, plessthan0.01), while non-progressive motile sperm (4.78 - 21.48%) and immotile sperm (164.38 - 178.38 %) percentage increased significantly ( plessthan0.01) in frozen-thawed semen compared to that of fresh semen. The post-thaw quality of semen of all three breeds was in acceptable range. The mean values of sperm velocity/kinematic parameters observed in frozen-thawed semen of Gir, Surti and Murrah bulls, based on total motile sperms, viz., average path velocity, curvilinear velocity, straight line velocity, linearity, straightness, beat-cross frequency, amplitude of lateral head displacement, dancing frequency and dancing mean decreased significantly by 13.70 - 17.79; 9.76 - 12.95; 13.28 - 21.90; 7.28 - 9.68; 4.36 - 7.79; 15.56 - 25.15; 8.78 - 10.50; 6.16 - 18.67 and 12.98 - 15.96 %, respectively, as compared to that of their fresh semen samples. However, wobbling index remained almost same for both fresh and frozen semen. All motility traits differed but none of kinematics/velocity traits differed significantly between breeds/species. The values of all velocity parameters for progressive motile sperms were higher than total motile sperms in all three breeds. The effect of freezing-thawing on velocity and kinematic attributes was much less compared to absolute sperm motility, and both the fresh and frozen-thawed sperms behaved identically with respect to their velocity and kinematics. The rapid progressive motile sperm in both fresh (r=0.41 to 0.92) and frozen-thawed (r=044 to 0.88) semen had significant correlations with most of their velocity traits, and the later were significantly and positively or negatively inter-related among each other in semen of all three breeds. It was therefore concluded that cryopreservation process significantly reduces the motility and kinematics attributes of bovine spermatozoa and, CASA analysis of fresh semen for motility and velocity traits could predict the post-thawed sperm motility and velocity/ kinematics of spermatozoa.

Download Full-text

The effects of Trolox on the quality of sperm from captive squirrel monkey during liquefaction in the extender ACP-118™

Zygote ◽

10.1017/s096719941800028x ◽

2018 ◽

Vol 26 (4) ◽

pp. 333-335 ◽

Cited By ~ 2

Author(s):

D. V. C. Almeida ◽

J. S. Lima ◽

D. L. Leão ◽

K. G. Oliveira ◽

R. R. Santos ◽

...

Keyword(s):

Plasma Membrane ◽

Squirrel Monkey ◽

Sperm Motility ◽

Membrane Integrity ◽

Plasma Membrane Integrity ◽

Time Periods ◽

Semen Extender ◽

Fresh Semen

SummaryThe aim of this study was to evaluate the effect of incubating semen for different periods (90, 270 or 450 min) with or without Trolox® (100 or 150 µM) on the quality of sperm from Saimiri collinsi. Sperm motility, vigour, and plasma membrane integrity (PMI) were evaluated in both fresh semen and semen incubated for different time periods, i.e. 90, 270 or 450 min of incubation. Supplementation of semen extender with Trolox® 100 µM improved sperm motility, vigour and PMI for up to 270 min of incubation.

Download Full-text