Comparing ethylene glycol with glycerol and with or without dithiothreitol and sucrose for cryopreservation of bull semen in egg-yolk containing extenders

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.

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Antibiotics for Bull Semen Frozen in Milk and Egg Yolk Extenders

Journal of Dairy Science ◽

10.3168/jds.s0022-0302(87)80306-5 ◽

1987 ◽

Vol 70 (11) ◽

pp. 2439-2443 ◽

Cited By ~ 5

Author(s):

K. Ahmad ◽

R.H. Foote ◽

M. Kaproth

Keyword(s):

Egg Yolk ◽

Bull Semen

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Freezability of Bull Semen in Centrifuged or Dialyzed and Reconstituted Buffered Egg Yolk Extenders

Journal of Dairy Science ◽

10.3168/jds.s0022-0302(67)87652-5 ◽

1967 ◽

Vol 50 (9) ◽

pp. 1481-1485 ◽

Cited By ~ 4

Author(s):

A.M. Yassen ◽

R.H. Foote

Keyword(s):

Egg Yolk ◽

Bull Semen

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Manganese Provides Antioxidant Protection for Sperm Cryopreservation that May Offer New Consideration for Clinical Fertility

Oxidative Medicine and Cellular Longevity ◽

10.4161/oxim.2.3.8804 ◽

2009 ◽

Vol 2 (3) ◽

pp. 152-159 ◽

Cited By ~ 24

Author(s):

Ranjna S. Cheema ◽

Amrit K. Bansal ◽

Gurmail Singh Bilaspuri

Keyword(s):

Protective Effect ◽

Semen Quality ◽

Maximum Level ◽

Egg Yolk ◽

Quality Parameters ◽

Vitro Fertilization ◽

Bull Semen ◽

Protein Leakage ◽

A Chain

Reactive oxygen species (ROS) are generated by sperm metabolism. While, ROS are required for maturation, capacitation and acrosome reaction, they also modify many peroxidable cellular compounds. There is production of ROS during cryopreservation and frozen spermatozoa are highly sensitive to lipid peroxidation (LPO). Antioxidants exert a protective effect on the plasma membrane of frozen bovine sperm preserving both metabolic activity and cellular viability. Manganese (Mn++) is proved to be a chain breaking antioxidant in biological system. Therefore, we examined the role of (Mn++) during cryopreservation of cattle bull semen. Semen was divided into four parts and cryopreserved in egg-yolk-citrate extender + glycerol (EYC-G), EYC-G + 100 µM of Mn++, EYC-G + 150 µM of Mn++and EYC-G + 200 µM of Mn++. After four hours of cooling and 24 hrs of freezing, the spermatozoa were examined for percentage motility, Hypo-osmotic swelling (HOS), LPO and protein leakage. Addition of manganese to the semen during cryopreservation showed a protective effect and accounted for an increase in semen quality parameters [percentage motility, HOS percent and decrease in malondialdehyde (MDA) production and protein leakage]. The effect of manganese on motility and HOS was non-significant (p < 0.05) in cooled spermatozoa but significant with 150 µM of Mn++in frozen-thawed spermatozoa. MDA production and protein leakage decreased to a significant and maximum level (p < 0.05) on addition of 200 µM of manganese. The addition of manganese to EYC-G dilutor will improve the quality/fertility of semen, which will result in improvement of in vitro fertilization and artificial insemination success rate.

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KUALITAS MOTILITAS DAN VIABILITAS SPERMATOZOA DARI SEMEN AFKIR SAPI LIMOUSIN PADA PENGENCER SUSU SKIM KUNING TELUR SITRAT DENGAN PENAMBAHAN BERBAGAI KADAR GLUKOSA

Ovozoa Journal of Animal Reproduction ◽

10.20473/ovz.v7i2.2018.96-101 ◽

2020 ◽

Vol 7 (2) ◽

pp. 96

Author(s):

Rudi Irvanto ◽

Hardijanto Hardijanto ◽

Widya Paramita ◽

Suherni Susilowati ◽

Tita Damayanti L ◽

...

Keyword(s):

Research Design ◽

Glucose Level ◽

Skim Milk ◽

Egg Yolk ◽

Spermatozoa Motility ◽

Bull Semen ◽

Randomized Design ◽

Completely Randomized Design

Quality of spermatozoa motility and viability from rejected limousin bull semen diluted with skim milk egg yolk sitrat added with various levels of glucose. Glucose level used were 0%, 0,5%, 1,5%, 2,5%, and 3,5%. Writer was using on four years old Limousine bull. Bull semen used in this research was bull rejected semen with bellow 70% motility. Semen observation was done at 0 hour, 24 hours and 48 hours. Research design used in this study was completely randomized design with faktorial pattern with 5 replicates. Highest result in motility this research was showed at 24 hours with 30% value in glucose 2,5% treatment and 48 hours with 10% value. The lowest result showed in glucose 0% treatment at 24 hours and 0% at 48 hours. Highest result in viability showed on glucose 2,5% treatment with 62,6% value at 24 hours and at 48 hours with 53,4% value. Lowest result in viability showed on glucose 0% treatment with 44,2% value at 24 hours and 31,4% value at 48 hours.

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