Evaluation of sperm motility across varying thawing temperatures in Jersy, Holstein Friesian, and Murrah Bulls

Rajesh  Gautam; Bhaju Ram  Mahato; Amrit  Shrestha; Suman Kumar  Singh; Muhammad  Adil; Syed Mubash Sher  Tajmir; Md. Mahfuzul  Haque

Rajesh Gautam Institute of Agriculture and Animal Science (IAAS), Tribhuvan University, Paklihawa Campus, Rupandehi, 32900, Nepal
Bhaju Ram Mahato Ministry of Agriculture and Livestock Development, Department of livestock Services, National Livestock Breeding Office, Pokhara, Nepal
Amrit Shrestha State key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Graduate School of Chinese Academy of Agricultural Sciences, China
Suman Kumar Singh Department of Veterinary Surgery, Medicine, Epidemiology and Public Health, Institute of Agriculture and Animal Science (IAAS), Tribhuvan University, Paklihawa Campus, Rupandehi, Nepal
Muhammad Adil Department of Clinical Medicine and Surgery, University of Agriculture, Faisalabad, Pakistan
Syed Mubash Sher Tajmir Lanzhou Veterinary Research Institute, Graduate School of Chinese Academy of Agricultural Sciences, China
Md. Mahfuzul Haque Department of Surgery and Theriogenology, Sylhet Agricultural University, Bangladesh

Keywords: Thawing, Semen analysis, Conception rate, Reproduction failures, Artificial insemination, cattle

Abstract

Background: The heating rate during the thawing of frozen semen significantly impacts the functional activation of mitochondria, which affects sperm motility assessment.

Methods: This study examined the effect of thawing temperature on the motility of spermatozoa in cryopreserved semen. A total of 240 semen straws (0.25 ml each) from 24 bulls (8 Jersey, 8 Holstein Friesian (HF), and 8 Murrah breed), aged 2 to 6 years, were used. Semen was collected, analyzed, processed, frozen, and stored in liquid nitrogen using a standard protocol with a tris-citrate-egg yolk extender. Samples were thawed for 30 seconds in a water bath at temperatures T1: 32°C, T2: 34°C, T3: 36°C, T4: 38°C, and T5: 40°C. Sperm motility, progressive motility, fast motility, slow motility, and immotile percentage of the frozen semen were evaluated. A computer-assisted semen analyzer (CASA) was used for analysis. The data was entered into MS-Excel and analyzed using analysis of variance (ANOVA), with the significance between treatments assessed using Duncan’s multiple range test (DMRT) post hoc test.

Results: Jersey semen showed the best motility percentage at T3 (p<0.05) (T1: 53.025±6.73, T2: 60.01±3.81, T3: 74.33±1.40, T4: 65.59±3.17, T5: 59.92±3.58). HF semen also showed the best motility percentage at T3 (p<0.05) (T1: 60.67±6.31, T2: 55.93±6.31, T3: 76.6±2.28, T4: 72.93±2.10, T5: 57.69±2.28). Murrah semen showed the best motility percentage at T4 (p<0.05) (T1: 56.63±3.93, T2: 58.58±4.34, T3: 77.09±1.59, T4: 82.72±4.03, T5: 72.87±4.00). Progressive motility (%) was highest at T3 for Jersey (52.41±2.97), T3 for HF (56.29±4.65), and T4 for Murrah (63.94±6.37) (p<0.05). Fast motility (%) was highest at T3 for Jersey (12.71±2.04), T3 for HF (18.43±1.63), and T4 for Murrah (12.589±2.74) (p<0.05). Slow motility (%) was lowest at T3 for Jersey (27.8±3.43), T3 for HF (34.23±2.72), and T4 for Murrah (31.32±4.72) (p<0.05). Immotility (%) was lowest at T3 for Jersey (25.73±1.37), T3 for HF (23.4±2.28), and T4 for Murrah (17.27±4.03) (p<0.05).

Conclusion: Thawing at T3 (36°C) enhances motility, progressive motility, and fast motility while reducing slow motility and immotile percentage in Jersey semen. Similarly, thawing at T4 (38°C) improves motility, progressive motility, and fast motility while reducing slow motility and immotile percentage in Murrah semen.

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