Impact of Non-physiological Incubation Temperatures on Spermatological and Functional of Thawed Buffalo Spermatozoa

Burcu Esin; Cumali Kaya

Article Details

Impact of Non-physiological Incubation Temperatures on Spermatological and Functional of Thawed Buffalo Spermatozoa

Authors:

Burcu Esin | Cumali Kaya

Publisher:

Gültekin ÖZDEMİR

Main Topics:

Veterinary Urology

Journal:

International Journal of Agriculture Environment and Food Sciences Vol.9 Issue 3, pp.759-767

Document Type:

Article

Publication Date:

9/2025

Document Language:

English

Document ID (Org No.):

2001354275

pISSN:

2618-5946

Keywords:

Abstract:

 Cryopreservation is a widely accepted technique for preserving male gametes; however, post-thaw incubation conditions may substantially affect sperm quality and function. This study aimed to investigate the influence of different post-thaw incubation temperatures on the spermatological and functional parameters of buffalo bull spermatozoa. Fifteen semen straws from the same bull were thawed at 37C for 30 seconds, pooled to eliminate inter-straw variation, and equally divided into three groups: control 37C, cold shock 4C, and thermal stress 45C. All samples were incubated for 30 minutes before evaluation. Sperm motility and kinematics were analyzed using a computer-assisted sperm analysis system CASA. Viability was assessed with eosin-nigrosin staining, plasma membrane integrity with the hypoosmotic swelling test, acrosome integrity via SpermBlue staining, and chromatin condensation using Toluidine Blue staining. Incubation temperature had a statistically significant effect on all examined parameters P0.05. Both the cold shock and thermal stress groups exhibited a significant decrease in motility and progressive motility, kinematic parameters VCL, VSL, VAP, viability, and membrane integrity when compared to the control group P0.05 to P0.001. Chromatin decondensation levels were significantly higher in both groups compared to the control, and acrosome integrity was significantly compromised. Furthermore, thermal stress induced a significantly decrease in progressive motility, chromatin integrity, and acrosomal structure compared to cold shock P0.001. In conclusion, post-thaw exposure to non-physiological temperatures was observed to negatively affect buffalo sperm quality, highlighting the importance of thermal regulation in post-thaw handling during assisted reproduction procedures. Further studies are needed to evaluate the effects on live fertility outcomes.

 Cryopreservation is a widely accepted technique for preserving male gametes; however, post-thaw incubation conditions may substantially affect sperm quality and function. This study aimed to investigate the influence of different post-thaw incubation temperatures on the spermatological and functional parameters of buffalo bull spermatozoa. Fifteen semen straws from the same bull were thawed at 37C for 30 seconds, pooled to eliminate inter-straw variation, and equally divided into three groups: control 37C, cold shock 4C, and thermal stress 45C. All samples were incubated for 30 minutes before evaluation. Sperm motility and kinematics were analyzed using a computer-assisted sperm analysis system CASA. Viability was assessed with eosin-nigrosin staining, plasma membrane integrity with the hypoosmotic swelling test, acrosome integrity via SpermBlue staining, and chromatin condensation using Toluidine Blue staining. Incubation temperature had a statistically significant effect on all examined parameters P0.05. Both the cold shock and thermal stress groups exhibited a significant decrease in motility and progressive motility, kinematic parameters VCL, VSL, VAP, viability, and membrane integrity when compared to the control group P0.05 to P0.001. Chromatin decondensation levels were significantly higher in both groups compared to the control, and acrosome integrity was significantly compromised. Furthermore, thermal stress induced a significantly decrease in progressive motility, chromatin integrity, and acrosomal structure compared to cold shock P0.001. In conclusion, post-thaw exposure to non-physiological temperatures was observed to negatively affect buffalo sperm quality, highlighting the importance of thermal regulation in post-thaw handling during assisted reproduction procedures. Further studies are needed to evaluate the effects on live fertility outcomes.

 Cryopreservation is a widely accepted technique for preserving male gametes; however, post-thaw incubation conditions may substantially affect sperm quality and function. This study aimed to investigate the influence of different post-thaw incubation temperatures on the spermatological and functional parameters of buffalo bull spermatozoa. Fifteen semen straws from the same bull were thawed at 37C for 30 seconds, pooled to eliminate inter-straw variation, and equally divided into three groups: control 37C, cold shock 4C, and thermal stress 45C. All samples were incubated for 30 minutes before evaluation. Sperm motility and kinematics were analyzed using a computer-assisted sperm analysis system CASA. Viability was assessed with eosin-nigrosin staining, plasma membrane integrity with the hypoosmotic swelling test, acrosome integrity via SpermBlue staining, and chromatin condensation using Toluidine Blue staining. Incubation temperature had a statistically significant effect on all examined parameters P0.05. Both the cold shock and thermal stress groups exhibited a significant decrease in motility and progressive motility, kinematic parameters VCL, VSL, VAP, viability, and membrane integrity when compared to the control group P0.05 to P0.001. Chromatin decondensation levels were significantly higher in both groups compared to the control, and acrosome integrity was significantly compromised. Furthermore, thermal stress induced a significantly decrease in progressive motility, chromatin integrity, and acrosomal structure compared to cold shock P0.001. In conclusion, post-thaw exposure to non-physiological temperatures was observed to negatively affect buffalo sperm quality, highlighting the importance of thermal regulation in post-thaw handling during assisted reproduction procedures. Further studies are needed to evaluate the effects on live fertility outcomes.

 Cryopreservation is a widely accepted technique for preserving male gametes; however, post-thaw incubation conditions may substantially affect sperm quality and function. This study aimed to investigate the influence of different post-thaw incubation temperatures on the spermatological and functional parameters of buffalo bull spermatozoa. Fifteen semen straws from the same bull were thawed at 37C for 30 seconds, pooled to eliminate inter-straw variation, and equally divided into three groups: control 37C, cold shock 4C, and thermal stress 45C. All samples were incubated for 30 minutes before evaluation. Sperm motility and kinematics were analyzed using a computer-assisted sperm analysis system CASA. Viability was assessed with eosin-nigrosin staining, plasma membrane integrity with the hypoosmotic swelling test, acrosome integrity via SpermBlue staining, and chromatin condensation using Toluidine Blue staining. Incubation temperature had a statistically significant effect on all examined parameters P0.05. Both the cold shock and thermal stress groups exhibited a significant decrease in motility and progressive motility, kinematic parameters VCL, VSL, VAP, viability, and membrane integrity when compared to the control group P0.05 to P0.001. Chromatin decondensation levels were significantly higher in both groups compared to the control, and acrosome integrity was significantly compromised. Furthermore, thermal stress induced a significantly decrease in progressive motility, chromatin integrity, and acrosomal structure compared to cold shock P0.001. In conclusion, post-thaw exposure to non-physiological temperatures was observed to negatively affect buffalo sperm quality, highlighting the importance of thermal regulation in post-thaw handling during assisted reproduction procedures. Further studies are needed to evaluate the effects on live fertility outcomes.

Article Details

Impact of Non-physiological Incubation Temperatures on Spermatological and Functional of Thawed Buffalo Spermatozoa

Authors:

Burcu Esin | Cumali Kaya

Publisher:

Gültekin ÖZDEMİR

Main Topics:

Veterinary Urology

Journal:

International Journal of Agriculture Environment and Food Sciences Vol.9 Issue 3, pp.759-767

Document Type:

Article

Publication Date:

9/2025

Document Language:

English

Document ID (Org No.):

2001354275

pISSN:

2618-5946

Buffalo spermatozoa

Cold shock

Post-thaw incubation

Sperm quality

Thermal stress

Issues

Article Details

Impact of Non-physiological Incubation Temperatures on Spermatological and Functional of Thawed Buffalo Spermatozoa

Authors: Burcu Esin | Cumali Kaya

Publisher: Gültekin ÖZDEMİR

Main Topics: Veterinary Urology

Journal: International Journal of Agriculture Environment and Food Sciences Vol.9 Issue 3, pp.759-767

Document Type: Article

Publication Date: 9/2025

Document Language: English

Document ID (Org No.): 2001354275

pISSN: 2618-5946

Buffalo spermatozoa

Cold shock

Post-thaw incubation

Sperm quality

Thermal stress

Issues

Authors:

Burcu Esin | Cumali Kaya

Publisher:

Gültekin ÖZDEMİR

Main Topics:

Veterinary Urology

Journal:

International Journal of Agriculture Environment and Food Sciences Vol.9 Issue 3, pp.759-767

Document Type:

Article

Publication Date:

9/2025

Document Language:

English

Document ID (Org No.):

2001354275

pISSN:

2618-5946