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Post-thaw Quality of Modified Dextran Swim-up Separated Bull’s Semen Fractions

Zannatul Maoya
Department of Surgery and Obstetrics, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh

Mofizul Islam
Department of Surgery and Obstetrics, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh

Aynul Hoque
Department of Surgery and Obstetrics, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh

Farida Yeasmin Bari
Department of Surgery and Obstetrics, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh

Nasrin Sultana Juyena
Department of Surgery and Obstetrics, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh

Abstract/ Executive Summary:

Development of alternative, less harmful and feasible sperm sexing procedures through swim-up technique are cost-effective and highly desirable for domestic animals, especially bulls. Hence, the study was planned to assess fresh and post-thaw sperm quality after dextran swim-up separation. Semen was collected from four bulls to evaluate sperm quality, and separated fractions were divided into two groups (10 and 15 min.) according to the time allowed for swim-up separation using dextran transition layer and later cryopreserved using Tris-based extender. It was observed that concentration of spermatozoa and progressive motility% significantly (P<0.05) and motility% insignificantly (P>0.05) decreased in all fractions of two groups in comparison to fresh whole semen. We found significantly (P<0.05) the highest concentration of spermatozoa in Fraction 1 in both time groups. No significant (P>0.05) difference was observed in different parameters except motility% of Fraction 4 of two groups. Notably, there was no significant (P>0.05) difference found in kinematic parameters of fractioned semen collected at 10- and 15-minute intervals. After freezing and thawing, motility% and progressive motility% significantly (P<0.05) decreased in all fractions irrespective of groups. The highest post-thaw motility (45.57±7.54) and progressive motility% (5.42±1.50) was observed in Fraction 2 and Fraction 4 of 15 min group, respectively. This preliminary study could help animal andrologist to design sex pre-selective semen preparation protocol in a cost-effective way.

Keywords: Spermatozoa, Swim-up, Cryopreservation

Location: Research Animal Farm, Department of Surgery and Obstetrics, Bangladesh Agricultural University (BAU), Mymensingh, Bangladesh

Start Date: 00-01-2017

End Date: 00-05-2019

Program Area: Animal Health and Management

Commodity: Bull, Animal

Objectives:

To evaluate the quality of fresh and frozen swim-up separated fractions concerning collection time by studying kinetic parameters of semen using CASA.

Methodology:

Research duration and place: The experiment was performed from January 2019 to May 2019 at Research Animal Farm, Department of Surgery and Obstetrics, Bangladesh Agricultural University (BAU). Experimental animals: Semen was collected from pure (100%) Holstein-Friesian (Harison), pure (100%) Sahiwal (Bahadur), 87.5% Holstein-Friesian crossbred (George) and 87.5% Sahiwal (Sultan) crossbred bulls. Experimental procedure: Modified human tubular fluid and 1% dextran solution were used as separation media. After semen collection, the routine evaluation was performed along with the kinetic velocities using a computer-assisted semen analyzer. Four fractions were collected following 10 minutes and 15 minutes intervals. Samples were grouped according to the time of collection. Each fraction was evaluated for its kinematic parameters and plasma membrane functional integrity. Tris-based Media was prepared using Tris (2.42%w/v), citric acid (1.36%w/v), fructose (1%w/v), 20% (v/v) egg yolk, glycerol 7% (v/v), penicillin (100,000 IU), streptomycin (100 mg) and distilled water. The media was added in an equal volume (1:1) with the separated fractions for cryopreservation in one step method. Preservation was performed at -1960C for further evaluation in 0.25 ml straws. After thawing, semen of each fraction was evaluated for the kinematic parameters along with the motility% and progressive motility% using CASA. Data analysis: Data was recorded in an excel worksheet obtained from CASA software. Then the statistical analysis was performed by paired t-test to find out the comparison in each parameter between the fresh and frozen condition of semen within a specific time interval with SPSS 20.0.

Source of Information: J Bangladesh Agril Univ 18(S1): 853–857, 2020 ISSN 1810-3030 (Print) 2408-8684 (Online)

Article Link (Online Journal): https://doi.org/10.5455/JBAU.25155

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

In this study, effects for all sperm quality parameters were studied within bulls and fractions. The concentration of spermatozoa and progressive motility% significantly (P<0.05) and motility% insignificantly (P>0.05) decreased in all fractions of two groups in comparison to fresh whole semen. After freezing and thawing, motility% and progressive motility% significantly (P<0.05) decreased in all fractions irrespective of groups. The dextran swim-up procedure is inexpensive and simple to perform. To maximize reproductive efficiency and the fertilizing ability of semen samples, a reduction in the number of spermatozoa used per insemination is an important objective in animal production. Therefore, we should design future studies to freeze swim-up separated fractions in an acceptable and feasible way to produce the desired number of male and female offspring. We strongly believe that more studies should confirm the data before recommendations for widespread use of the modified procedure used in this study.

Knowledge Management: Journal