Effect of graphene oxide as cryoprotectant on post-thaw sperm functional and kinetic parameters of cross bred (HF X Sahiwal) and Murrah buffalo (Bubalus bubalis) bulls

• Published in: Cryobiology 2022-06, Vol.106, p.102-112
• Main Authors: Singh, Prahlad, Bedi, Munish Kumar, Singhal, Sumit, Singh, Ashwani Kumar, Kumar, Ajeet, Honparkhe, Mrigank
• Format: Article
• Language: English
• Subjects: Buffalo; CASA; Cattle; Cryoprotectant; Graphene oxide; Spermatozoa
• ISSN: 0011-2240; 1090-2392
• DOI: 10.1016/j.cryobiol.2022.03.002

Graphene oxide (GO) greatly suppresses the growth and recrystallization by curving the hexagonal shape of ice crystals. Study was conducted to evaluate effect of GO as cryoprotectant in semen extender for augmenting sperm viability in dairy (cattle and buffalo) animals. In experiment one, semen was extended with TRIS Egg Yolk Glycerol (TEYG) extender supplemented with different concentrations of GO: 0.0125, 0.25, 0.5, 0.1 and 0.2 mg ml−1. Freezing of semen samples was conducted at 30 °C min−1 from temperature drop from 4 °C to −15 °C and −15 °C to – 60 °C followed by 50 °C min−1 from – 60 °C to −140 °C, and the semen straws were plunged in liquid nitrogen. Second experiment evaluated the performance of TEYG extender supplemented with combinations of GO (G05 as 0.05 and G10 as 0.1 mg ml−1) and glycerol (T48 as 4.8 and T64 as 6.4%) in four groups as G05T48, G05T64, G10T48 and G10T64. Freezing rates of 30 °C min-1[Protocol (PRT) I], 40 °C min−1 (PRT II) and 50 °C min−1 (PRT III) in the critical temperature fall zone of −15 °C to −60 °C were evaluated for semen extender supplemented with glycerol 6.4% and GO 0.05 mg ml−1 in the third experiment. Cattle (n = 3) and buffalo (n = 3) bulls were chosen for the study taking six ejaculates per bull per treatment. Post-thaw sperm motility, membrane integrity, viability and abnormalities were observed by means of CASA, Hypo-osmotic swelling test (HOST), Eosin-Nigrosin stain and Rose Bengal stain procedures, respectively. Post-thaw total motility (TM), progressive motility (PM), VCL, VSL, VAP, HOST response and viability increased significantly in extender with GO concentrations of 0.1 and 0.05 mg ml−1 as compared to control. Per cent abnormalities were significantly (p