Cryoprotection synergism between glycerol and dimethyl sulfoxide improves the mitochondrial transmembrane potential, plasmalemma, acrosomal and DNA integrities, and in vivo fertility of water buffalo (Bubalus bubalis) spermatozoa

Abstract

The objective of the study was to devise a cryoprotection synergism between glycerol and dimethyl sulfoxide (DMSO) for water buffalo spermatozoa. Additionally, the effect of best evolved concentrations of glycerol and DMSO in extender was assessed on in vivo fertility of buffalo spermatozoa. Ejaculates (n = 30) were equally distributed into five aliquots; first aliquot was diluted at 37°C in extender having 7 % glycerol (control); second aliquot was diluted at 37°C as well as at 4°C in extender having 3.5 % DMSO (Group 1); third aliquot was diluted at 37°C in extender having 3.5 % glycerol and then at 4°C in extender having 3.5 % DMSO (Group 2); fourth aliquot was diluted at 37°C in extender having 3.5 % DMSO and then at 4°C in extender having 3.5 % glycerol (Group 3); fifth aliquot was diluted in extenders having 1.75 % glycerol and 1.75 % DMSO at 37 as well as at 4°C (Group 4). At post thawing, sperm progressive motility (%), rapid velocity (%), average path velocity (lm/s), curved line velocity (lm/s), in vitro longevity (%), structural and functional integrity of plasmalemma (%), mitochondrial transmembrane potential (%) and viable sperm with intact acrosome (%) were higher (P \ 0.05) in Group 4 compared to other treatment groups and control. Regarding sperm DNA integrity (%); it was higher (P \ 0.05) in Group 4 compared to Group 1, 3 and control. The in vivo fertility (%) of buffalo spermatozoa was significantly higher with Group 4 compared to control (69.45 vs. 59.81). In conclusion, synergism exists between glycerol and DMSO (Group 4) in improving the quality and in vivo fertility of cryopreserved water buffalo spermatozoa.

Addition of pomegranate juice (Punica granatum) in tris‐based extender improves post‐thaw quality, motion dynamics and in vivo fertility of Nili Ravi buffalo (Bubalus bubalis) bull spermatozoa

“…The improvement in fertility rate in Nili Ravi buffaloes is imperative for the improvement of reproductive and productive efficiency (Shah, Andrabi, Ahmed, & Qureshi, 2016).…”

Effect of Synergism Between Carboxylated Poly-l-Lysine and Glycerol on Freezability of Nili-Ravi Buffalo (Bubalus bubalis) Semen

“…21 Sperm chromatin integrity. Sperm chromatin integrity of buffalo spermatozoa was assessed by acridine orange test modified by Shah et al 11 and formerly described by Martins et al 22 A semen smear of each experimental extender was prepared on microscopic slides (37°C), allowed to air dry at room temperature, and fixed in Carnoy's solution (glacial acetic acid and methanol, 3:1 proportion; Merck) for 2 hours at room temperature. After fixation, air-dried microscopic glass slides were placed at 75°C for 5 minutes in Tampon solution (15 mmol/L Na 2 HPO 4 $7H 2 0; 80 mmol/L citric acid; pH 2.5; Merck).…”

“…Studies are also being conducted to investigate nonpermeating and permeating cryoprotectants for their replacement with better agents in terms of improved freeze-thaw quality and reduced oxidative stress. [9][10][11]34,35 Glycerol has been used traditionally as a permeating cryoprotectant for >70 years, inevitably associated with higher cytotoxicity and less cryoprotectant ability compared to modern day cryoprotectants like CPLL. 12 CPLL has low toxicity compared to other cryoprotectants.…”

“…9,10 In this regard, different studies have been conducted to replace or reduce the level of glycerol in semen extender with other permeating cryoprotectants such as dimethyl sulfoxide. [9][10][11] Unfortunately, dimethyl sulfoxide has not gained popularity in semen cryopreservation industry due to contradictory results and its toxic and osmotic effects on buffalo spermatozoa. [9][10][11] Therefore, exploration of alternate cryoprotectants with low cytotoxicity is needed.…”

Association of SNPs in GnRH gene with sperm quality traits of Chinese water buffalo

“…The straws were directly immersed in liquid nitrogen and stored in a cryogenic container at 196°C for 5–7 days until the time of post‐thaw analysis (Ramires Neto et al, ). The straws of each randomly chosen buffalo were thawed in a thermostat water bath cauldron at 37°C for 30 s (Shah, Andrabi, Ahmed, & Qureshi, ). The motilities of the post‐thaw cryopreserved sperms were observed through a computer monitor connected to a camera mounted onto a phase‐contrast microscope (Olympus‐BX40, Minitüb) at 200× or 400× magnification and evaluated immediately (Yang et al, ).…”