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Title: Effect of Various Concentration of Glycerol on Post Thaw Quality of Holstein-Friesian Bull Semen
Authors: Sakhi Bakhsh, Hamzo Khan Kumbhar, Rameez Raja, Hubdar Ali Kaleri, Ghulam Mustafa Solangi, Raza Ali Mangi, Zahid Ali Mangrio, Mohammad Naseem, Zakir Hussain Bozdar, Ali Anwar, Mohammad Arif, Deepesh Kumar Bhuptani, Kashif Malik, Gulzar Ahmed
Journal: Journal of Agriculture and Veterinary Science
Publisher: Zoo Botanica (SMC Pvt.) Ltd.
Country: Pakistan
Year: 2023
Volume: 2
Issue: 2
Language: English
Keywords: SemenGlycerolPost thaw qualityHolstein Frisian
The aims of study were to compare the glycerol concentration in Tris, Citric acid Egg yolk diluents on the quality of Holstein Friesian frozen semen. Three Holstein Friesian bulls were maintained to collect the 45 semen samples. Total of 15 ejaculates sample were collected from each bull. Fresh semen was evaluated for volume, color, pH, concentration and motility. Every collection of 3 ejaculate was pooled and examined. Each pooled semen sample was divided in to three experimental extenders A, B, C, glycerol was used as diluents at 7, 6, 5%, Pencillin (1000 I.U/ml) and streptomycin (1000g/ml) were added to each diluent. Immediately after completion of the semen extension process, the post dilution motility percentage of each extender was noted. After 24 hours of freezing and storage liquid nitrogen -196 oC, three sample straws from each glycerol concentration were taken out and thawed in water bath at 370C for 15 seconds. After thawing the semen was tested by sperm quality analyzer SQA vb for bull post thaw motility % , progressive motility %, average pathway velocity µm/s, straight-line velocity µm/s, curvilinear velocity µm/s varied significantly between 5, 6 and 7% glycerol concentration to treatment p>0.05. Results of the experiment demonstrated that post thawed sperm motility in the semen preserved with 7% glycerol higher than other levels of glycerol, PTM (49.64%), PM (34.76%), VAP (94.92 um/s), VSL(85.36 um/s), VCL(165.42 um/s). It was concluded that 7% glycerol is recommended for freezing media regarding with best result on post-thaw quality of Holsteins-Friesian bull semen.
To compare the effect of different glycerol concentrations (7%, 6%, and 5%) in Tris, Citric acid Egg yolk diluents on the post-thaw quality of Holstein Friesian bull semen.
Semen samples were collected from three Holstein Friesian bulls. Fresh semen was evaluated for volume, color, pH, concentration, and motility. Pooled semen samples were divided into three experimental extenders with varying glycerol concentrations (7%, 6%, 5%), along with Penicillin and Streptomycin. Post-dilution motility was recorded. After 24 hours of freezing at -196°C, semen straws were thawed and tested for post-thaw motility, progressive motility, and various velocity parameters using a Sperm Quality Analyzer (SQA vb). Data was analyzed using ANOVA.
graph TD
A[Semen Collection from 3 Bulls] --> B[Fresh Semen Evaluation];
B --> C[Divide Semen into 3 Extenders7%, 6%, 5% Glycerol];
C --> D[Freezing and Storage];
D --> E[Thawing of Semen Straws];
E --> F[Post-Thaw Sperm Quality Analysis];
F --> G[Statistical AnalysisANOVA];
G --> H[Conclusion and Recommendation];
The results indicate that 7% glycerol is optimal for preserving the post-thaw quality of Holstein-Friesian bull semen. This concentration appears to minimize freezing and thawing stresses, leading to better sperm viability and motility. The study also noted variations in semen parameters among individual bulls, which could be attributed to physiological differences.
The study demonstrated that glycerol concentration significantly affected post-thaw sperm quality. Semen preserved with 7% glycerol showed significantly higher post-thaw motility (49.64%), progressive motility (34.76%), average pathway velocity (94.92 µm/s), straight-line velocity (85.36 µm/s), and curvilinear velocity (165.42 µm/s) compared to 5% and 6% glycerol concentrations.
A glycerol concentration of 7% is recommended for freezing media to achieve the best results in terms of post-thaw quality of Holstein-Friesian bull semen.
1. Glycerol Concentrations Tested: The study tested three glycerol concentrations: 7%, 6%, and 5%.
2. Number of Bulls: Three Holstein Friesian bulls were used for semen collection.
3. Post-Thaw Motility with 7% Glycerol: The study reported a post-thaw motility of 49.64% for semen preserved with 7% glycerol.
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