Influence of thawing temperature on sperm motility, structure, and metabolism of frozen bovine semen

Abstract

The heating rate used during semen thawing plays an important role in reducing structural and functional damage to spermatozoa. In this study, we evaluated the influence of thawing temperature on semen quality, reactive oxygen species (ROS) production, and mitochondrial activity of cryopreserved bovine semen. A total of 195 straws of 0.5 mL from five Holstein Friesian bulls were used (39 straws per bull). Samples underwent 8 to 22 years of storage; they were processed under a standard protocol with tris-egg yolk and stored in liquid nitrogen. Samples were thawed for 30 seconds in a water bath at T1: 36 °C, T2: 38 °C or T3: 40 °C. Sperm motility and kinematics, morphology, structural membrane integrity (SMI), functional membrane integrity (FMI), acrosome integrity (AI), ROS, and mitochondrial membrane potential (ΔΨM) of post-thawing bovine sperm were evaluated. Generalized linear models were fitted to the data. Each model included the effects of bull, storage time, and treatment. The Shapiro-Wilk test was used to assess data normality, and means were compared using the Tukey test. T2 and T3 showed better results for sperm motility and kinematic parameters, SMI (%) (T1 41.9 ± 2.3; T2 45.7 ± 1.9; T3 47.4 ± 2.8), ROS (RFU/min) (T1 0.026 ± 0.007; T2 0.032 ± 0.001; T3 0.031 ± 0.001) and high-ΔΨM (RFU x 103) (67.1± 0,4; 71.3 ± 0.4; 74.2 ± 0.4) (P < 0.05). However, T1 had higher FMI (39.3 ± 2.3) than T2 (34.0 ± 1.9) (P < 0.05), though not significantly (P > 0.05) different from T3 (38.4 ± 2.2). Thawing temperatures of 38 °C and 40 °C increases motility, kinetics, membrane integrity, mitochondrial activity and ROS of cryopreserved bovine semen, compared with more conventional thawing at 36 °C.

Combination of nanoparticle green tea extract in tris-egg yolk extender and 39 °c thawing temperatures improve the sperm quality of post-thawed Kacang goat semen

“…However, the thawing of semen decreases the plasma membrane integrity, causes an early acrosomal reaction, and reduces sperm quality (Zenteno et al, 2023). Higher thawing temperature than optimum increased acrosomal damage, a rise in sperm metabolic rate (Borah et al, 2015), mitochondrial activity to produce more ATP, and the consequence is higher ROS production (Khalil et al, 2018), followed by hyperactivity and induce sperm death (Zenteno et al, 2023). In this study, the temperature of 39ºC result in an improvement in sperm quality than thawing at 37ºC.…”

“…The thawing at 37 °C 20 s to 70 °C 12 s has not affected the midpiece and sperm head morphometry (Demirhan et al, 2020). However, the thawing of semen decreases the plasma membrane integrity, causes an early acrosomal reaction, and reduces sperm quality (Zenteno et al, 2023). Higher thawing temperature than optimum increased acrosomal damage, a rise in sperm metabolic rate (Borah et al, 2015), mitochondrial activity to produce more ATP, and the consequence is higher ROS production (Khalil et al, 2018), followed by hyperactivity and induce sperm death (Zenteno et al, 2023).…”

Frozen Semen Quality of Simmental Cattle in Various Commercial Diluents

“…The decrease can be explained by the freezing process and reduction in temperature. Membrane damage caused by extreme temperature drops occurs due to the shrinking of the fluid in the membrane (Zenteno et al 2023). Swelling and shrinking are sperm responses to osmotic changes that can cause cell death or significant loss of membrane integrity (Mughal et al 2018).…”

Short‐term preservation of canine sperm‐binding ability and other metrics using the INRA‐96 in comparison to Tris‐egg yolk extender

“…In contrast, in that previous study, samples were rewarmed at 37°C for 10 min, whereas in the current study, this step was performed at the same temperature for only 1 min. Nothling and Schuttleworth (2005) observed that a 30-s warming protocol at 37°C is sufficient for the samples to reach this temperature, and Zenteno et al (2023) argue that sample warming should be done quickly to reduce structural and functional damage caused by osmotic and oxidative stress that can occur during slow warming and cellular metabolism reactivation.…”