Functional and Flow Cytometric Analysis of Buffalo Cryopreserved Spermatozoa: Comparison of Different Breeds and Incubation Times

Document Type : Original Article

Authors

1 Department of Anatomical Sciences, Faculty of Medicine, Urmia University of Medical Sciences, Urmia, Iran.

2 Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, Selcuk University, Konya, Turkey

3 Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

4 Department of Anatomical Sciences, Faculty of Medicine, Guilan University of Medical Sciences, Rasht, Iran

5 Department of Urology, Selcuk University School of Medicine, Konya, Turkey

6 Department of Medical Services and Techniques, Denizli Vocational School of Health Services, Pamukkale University, Denizli, Turkey

7 Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.

8 Buffalo Breeding and Training Extension Center, Jabal, Urmia, Iran

9 Department of Animal Science, Faculty of Veterinary Medicine, Selcuk University, Konya, Turkey

10 Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran Reproductive Epidemiology Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

Abstract

Background: The purpose of this research was to compare the functional parameters of frozen-thawed Iranian Azari
buffalo spermatozoa with imported semen samples of Italian Mediterranean buffalo (IMB) after the thawing process
and 4 hours of incubation.
Materials and Methods: In this experimental study, a total of twenty-four ejaculates from four Iranian Azari buffalo
bulls were collected. Semen samples were diluted in AndroMed extender at a concentration of 50×106 spermatozoa/
ml. The diluted samples were filled in 0.5 ml straws and were frozen in a programmable freezer. For imported semen
samples, twenty-four samples of four IMB were used, which were diluted in AndroMed extender and frozen by the same
procedure. Frozen-thawed sperm motion patterns, mitochondrial activity, membrane integrity, DNA integrity, reactive
oxygen species (ROS), and apoptosis status were evaluated immediately after thawing and 4 hours of incubation.
Results: Post-thawed sperm motility, progressive motility (PM), mitochondrial activity, membrane integrity were
significantly higher in imported semen samples in compare with Iranian Azari buffalo. After 4 hours of incubation,
sperm velocity patterns were superior in Iranian Azari semen samples. Moreover, the percentage of sperm cells with
un-damaged DNA was higher in Iranian semen samples compared to imported samples at the time 0 of incubation.
Following 4 hours of incubation, a significant increase in intracellular ROS level leads to reduced membrane integrity,
mitochondrial activity, and DNA integrity in both buffalo breeds. At time 4, Iranian samples showed significantly
lower apoptosis and higher dead spermatozoa compared to imported semen samples.
Conclusion: Our study showed that the post-thawed quality of Iranian Azari buffalo semen was comparable with imported samples after 4 hours of incubation. Further investigations are recommended to assess the in vitro and in vivo
fertility rate of both buffalo breeds.

Keywords


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