The present study aimed to study the effect of cryopreservation on the membrane, acrosome, DNA ,and mitochondrial integrities of buffalo bull spermatozoa. Two adult buffalo bulls were enrolled in this study. Semen samples were collected once weekly from each bull over a period of six successive weeks by means of artificial vagina. Samples were processed for cryopreservation. Individual motility, mitochondrial activity as well as membrane, acrosomal and DNA integrities of fresh and frozenthawed spermatozoa were evaluated. Results showed that the procedures applied for freeze-thawing resulted in significant (P ≤ 0.001) reduction in sperm motility, sperm membrane, acrosome, mitochondrial and DNA integrities by 26.67, 29.17, 17.66, 40 and 4.11%, respectively. In conclusion, the process of freezing and thawing significantly reduced semen quality of buffalo bull in terms of various functional parameters including motility, membrane, acrosome, DNA and mitochondrial integrities. Keywords Cryopreservation, Buffalo, Spermatozoa, DNA, Mitochondria
IntroductionArtificial insemination (AI) is the most important reproductive biotechnology that can be applied in a large scale for dissemination of superior genetic material of males. (Singh & Balhara, 2016) The quality of frozen semen is a major determinant to the successful of AI program. Accurate evaluation of fertility of bulls used for AI is of utmost importance since a single ejaculate provides several insemination doses (up to 500 doses. (Pesch & Hoffmann., 2007), and influences the reproductive potential of a herd. (Rodriguez-Martinez & Larsson., 1998) Cryopreservation is a non-physiological and complex method that involves a high level of adaptation of biological cells to the thermal and osmotic shocks that occur both during the dilution, cooling-freezing and during the thawing procedures. (Holt, 2000) During cryopreservation, freeze-thaw damages are unavoidable that result in reduced semen quality. (Yoshida, 2000) The process of semen cryopreservation is stressful and around 40%-50% of sperm lose their motility Article info: Citation: Mahrous A., Answer A., El-Badry D. & Zain A. (2022) Cryopreservation-induced changes in buffalo spermatozoa with special reference to their DNA, membrane, acrosomal and mitochondrial integrities.