Is addition or removal of seminal plasma able to compensate for the dilution effect of buffalo semen?

Abstract

Summary The present study aimed to compensate dilution effect using additional seminal plasma (SP) in conventional (80 million (M) spermatozoa/ml) dose and low spermatozoa/dose (8M spermatozoa/ml). We also attempted to confirm whether removal of SP before the extension of ejaculates affects post‐thaw sperm quality of buffalo semen. For this, semen ejaculates (N = 15) were divided into four groups: control (CON), removal of SP by centrifugation (NSP), resuspension of the centrifuged semen pellet into SP (CEN) and extra supplementation of SP (ESP). All groups were diluted into two different semen doses to 20 and 2M spermatozoa/0.25 ml using tris egg yolk extender and subsequently cryopreserved. We found that neither addition nor removal of SP affected sperm motility, kinematics, longevity, mitochondrial superoxide production and high mitochondrial membrane potential (MMP). Further, the addition or removal of SP was not able to compensate dilution effect in 2M groups resulting in a significantly (p < .05) reduction in sperm motility, kinematics, sperm longevity, membrane integrity, MMP, and an increase production of mitochondrial superoxide. In conclusion, it appears that role of SP in the sperm cryopreservation process is insignificant.

Effect of mitochondria‐targeted antioxidant on the regulation of the mitochondrial function of sperm during cryopreservation

“…Superoxide anion is the principal ROS produced by mitochondria. The MitoSOX‐based assay has been used to identify sperm population that generated high superoxide after the freeze–thaw process due to the ability of MitoSOX to permeate live cells where it selectively targets mitochondria (Arjun et al, 2021; Dalal, Chandolia, Jan, et al, 2020; Patil et al, 2020). It is quickly oxidized by superoxide but not by other ROS.…”

“…The JC‐1 dye was used to estimate MMP as performed by Arjun et al (2021). The working solution of JC‐1 (1.5 mM in DMSO) was mixed with a semen sample to make a final concentration of 2 μM of JC‐1.…”

“…Sperm mitochondrion is one of the major susceptible organelles that get damaged during cryopreservation, which could be responsible for the reduction of sperm motility after a freeze–thaw process (Arjun et al, 2021; Kumar et al, 2021). The compromised mitochondrion is accountable for reactive oxygen species (ROS) generation through the production of superoxide in the electron transport chain system (Koppers et al, 2008).…”

Effect of over dilution of semen with tris extender on motion and functional attributes of bull spermatozoa during cryopreservation

“…However the proportions of moribund spermatozoa, dead spermatozoa, live acrosome reacted (LAR) spermatozoa, dead acrosome intact (DAI) spermatozoa and capacitated spermatozoa increased ( p < 0.05) when semen was diluted from 20 to 5 M. We found that sperm protamine deficiency and sperm lipid peroxidation were not affected by dilution. The semen dilution significantly affected the proportion of HOS responsive spermatozoa when buffalo semen was diluted to 2 million spermatozoa (Arjun et al, 2021). In mammals, the dynamics of sperm chromatin are greatly determined by protamination of sperm DNA (Talbert & Henikoff, 2010; Ward, 2010).…”

Protective effects of different doses of MitoQ separately and combined with trehalose on sperm function and antioxidative status of cryopreserved Markhoz goat semen

“…and rat [24] kidney after cold preservation. MitoQ could cross the rat blood-testis barrier and protect against rodent testicular injury by promoting the expression of mitochondrial dynamics proteins [44], activating antioxidant enzymes, and inhibiting apoptosis indicators [19] .The addition of MitoQ at various nanomolar concentrations in extender lowered the production of ROS and lipid peroxidation and improved sperm quality in frozen-thawed semen of catfish [13], human [21], buffalo [6], and rooster [25]. In buffalo semen, MitoQ indicated more cryotolerance than Resveratrol as a cytosolic antioxidant [38].…”