Effect of freezing rate on goat sperm morphology and DNA integrity

This study investigates the effect of freezing rates on the spermatological parameters of frozen and thawed Saanen goat semen. Equilibrated semen was frozen at 4 different freezing rates from +5 °C to -150 °C (G10: 10 °C/min, G12: 12 °C/min, G15: 15 °C/min, and G24: 24 °C/min) and stored in liquid nitrogen until use. Semen samples were examined for sperm motility, defective acrosomes (FITC-PSA), and DNA integrity using TUNEL after dilution with extender A at equilibration and postthaw stage. There was no significant difference among the freezing stages in terms of DNA fragmentation (P > 0.05). DNA integrity was partially affected by the freezing rate. The increase of freezing rate from 10 °C/min to 24 °C/min between +5 °C and -150 °C resulted in higher postthaw DNA damage. The study found that the freeze-thawing process is detrimental to postthawed goat semen motility (P < 0.05), acrosome integrity (P < 0.05), and DNA integrity (P > 0.05). Although the freezing rates used in the present study had no effect on postthaw sperm motility and acrosome integrity (P > 0.05), sperm DNA integrity was affected.

Effect of freezing rate on goat sperm morphology and DNA integrity

This study investigates the effect of freezing rates on the spermatological parameters of frozen and thawed Saanen goat semen. Equilibrated semen was frozen at 4 different freezing rates from +5 °C to -150 °C (G10: 10 °C/min, G12: 12 °C/min, G15: 15 °C/min, and G24: 24 °C/min) and stored in liquid nitrogen until use. Semen samples were examined for sperm motility, defective acrosomes (FITC-PSA), and DNA integrity using TUNEL after dilution with extender A at equilibration and postthaw stage. There was no significant difference among the freezing stages in terms of DNA fragmentation (P > 0.05). DNA integrity was partially affected by the freezing rate. The increase of freezing rate from 10 °C/min to 24 °C/min between +5 °C and -150 °C resulted in higher postthaw DNA damage. The study found that the freeze-thawing process is detrimental to postthawed goat semen motility (P < 0.05), acrosome integrity (P < 0.05), and DNA integrity (P > 0.05). Although the freezing rates used in the present study had no effect on postthaw sperm motility and acrosome integrity (P > 0.05), sperm DNA integrity was affected.

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  • Cooter PZ, Goolsby HA, Prien SD. Preliminary evaluation of a unique freezing technology for bovine spermatozoa cryopreservation. Reprod Dom Animal 2005; 40: 98–99.
  • Lemma A. Effect of cryopreservation on sperm quality and fertility. In: Manafi M, editor. Artificial Insemination in Farm Animals. Rijeka, Croatia: InTech; 2011. pp. 191–216.
  • Kumar S, Millar JD, Watson PF. The effect of cooling rate on the survival of cryopreserved bull, ram and boar spermatozoa: a comparison of two controlled rate cooling machines. Cryobiology 2003; 46: 246–253.
  • Kasimanickam R, Pelzer KD, Kasimanickam V, Swecker WS, Thatcher CD. Association of classical semen parameters, sperm DNA fragmentation index, lipid peroxidation and antioxidant enzymatic activity of semen in ram-lambs. Theriogenology 2006; 65: 1407–1421.
  • Amman RP, Hammerstedt RH. In vitro evaluation of semen quality: an opinion. J Androl 1993; 14: 397–406.
  • Larson KL, DeJonge CJ, Barnes AM, Jost, LK, Evenson DP. Sperm chromatin structure assay parameters as predictors of failed pregnancy following assisted reproductive techniques. Hum Reprod 2000; 15: 1717–1722.
  • Barroso G, Morshedi M, Oehninger S. Analysis of DNA fragmentation, plasma membrane translocation of phosphatidylserine and oxidative stress in human spermatozoa. Hum Reprod 2000; 15: 1338–1344.
  • Hammadeh ME, Askari A, Georg T, Rosenbaum P, Schmidt W. Effect of freeze-thawing procedure on chromatin stability, morphological alteration and membrane integrity of human spermatozoa in fertile and subfertile men. Int J Androl 1999; 22: 155–162.
  • Paasch U, Sharma RK, Gupta AK, Grunewald S, Mascha EJ, Thomas AJ Jr, Glander HJ, Agarwal A. Cryopreservation and thawing is associated with varying extent of activation of apoptotic machinery in subsets of ejaculated human spermatozoa. Biol Reprod 2004; 71: 1828–1837.
  • Nur Z, Zik B, Ustuner B, Tutuncu S, Sagirkaya H, Ozguden CG, Gunay U, Dogan I. Effect of freezing rate on acrosome and chromatin integrity in ram semen. Ankara Üniv Vet Fak Derg 2011; 58: 267–272.
  • Fraser L, Strzezek J. Effects of freezing–thawing on DNA integrity of boar spermatozoa assessed by the neutral comet assay. Reprod Dom Anim 2005; 40: 530–536.
  • Nur Z, Zik, B, Ustuner B, Sagirkaya H, Ozguden CG. Effects of different cryoprotective agents on ram sperm morphology and DNA integrity. Theriogenology 2010; 73: 1267–1275.
  • Batista M, Ninõ T, Alamo D, Castro N, Santana M, Gonzalez F, Cabrera F, Gracia A. Successful artificial insemination using semen frozen and stored by an ultrafreezer in the Majorera goat breed. Theriogenology 2009; 71: 1307–1315.
  • Ashrafi I, Kohram H, Naijian H, Bahreini M, Mirzakhani H. Effect of controlled and uncontrolled cooling rate on motility parameters of cryopreserved ram spermatozoa. Afr J Biol 2011; 10: 8965–8969.
  • Ustuner B, Gunay U, Nur Z. Effect of seminal plasma, egg yolk, and season on the freezability of Saanen buck semen. B Vet I Pulawy 2009; 53: 369–374.
  • Zhao BT, Han D, Xu CL, Luo MJ, Chang ZL, Tan JH. Protocol optimization for long-term liquid storage of goat semen in a chemically defined extender. Reprod Dom Anim 2009; 44: 865–872.
  • López-Fernández C, Johnston SD, Gosálbez A, Gosálvez J. Seasonal changes in sperm DNA fragmentation of Murciano- Granadina goats: the compelling case for dynamic assessment. Small Rum Res 2011; 100: 50–53.
  • Watson PF. AI and the preservation of semen. In: Lammingi GE, editor. Marshall’s Physiology of Reproduction. London, UK: Churchill Livingstone; 1990. pp. 747–869.
  • Barbas JP, Mascarenhas RD. Cryopreservation of domestic animal sperm cells. Cell Tissue Bank 2009; 10: 49–62.
  • Dorado J, Hidalgo M, Muñoz A, Rodríguez I. Assessment of goat semen freezability according to the spermatozoa characteristics from fresh and frozen samples. Anim Reprod Sci 2009; 112: 150–157.
  • Blanco JM, Gee G, Wildt DE, Donoghue AM. Species variation in osmotic, cryoprotectant, and cooling rate tolerance in poultry, eagle, and peregrine falcon spermatozoa. Biol of Reprod 2000; 63: 1164–1171.
  • Bittencourt RF, Ribeiro Filho AL, Alves SGG, Biscarde CE, Vasconcelos MF, Oba E. Cryopreservation of goat semen: the effect of freezing rate and equilibration time. Ciência Animal 2007; 17: 75–82 (in Portuguese with English abstract).
  • Medeiros CMO, Forell F, Oliveria ATD, Rodrigues JL. Current status of sperm cryopreservation: why isn’t it better? Theriogenology 2002; 57: 327–344.
  • Choe CY, Kim JG, Cho SR, Son DS, Kim YK, Balasubramanian, S, Choe SY, Rho GJ. Influence of seasons, extenders, slow and rapid freezing on seminal characters in Korean native bucks. Reprod Dom Anim 2006; 41: 55–60.
  • Frankel TE, Theisen DD, Guthrie HD, Welch GR, Woods LC. The effect of freezing rate on the quality of striped bass sperm. Theriogenology 2013; 79: 940–945.
  • Martorana K, Klooster K, Meyers S. Suprazero cooling rate, rather than freezing rate, determines post thaw quality of rhesus macaque sperm. Theriogenology 2013; 81:381–388.
  • Memon AA, Wahid H, Rosnina Y, Goh YM, Ebrahimi M, Nadia FM. Effect of ascorbic acid concentrations, methods of cooling and freezing on Boer goat semen cryopreservation. Reprod Dom Anim 2013; 48: 325–330.
  • Hafez ESE. Semen evaluation. In: Hafez ESE, editor. Reproduction in Farm Animals. 6th ed. Philadelphia: Lea & Febiger; 1993. pp. 405–423.
  • Benchaib M, Braun V, Lornage, J, Hadj S, Salle B, Lejeune H, Guerin JF. Sperm DNA fragmentation decreases the pregnancy rate in an assisted reproductive technique. Hum Reprod 2003; 18: 1023–1028.
  • Donnelly ET, McClure N, Lewis SE. Cryopreservation of human semen and prepared sperm: effects on motility parameters and DNA integrity. Fertil Steril 2001; 76: 892–900.
Turkish Journal of Veterinary and Animal Sciences-Cover
  • ISSN: 1300-0128
  • Yayın Aralığı: 6
  • Yayıncı: TÜBİTAK
Sayıdaki Diğer Makaleler

Effect of dietary seaweed (Ulva lactuca) supplementation on growth performance of sheep and on in vitro gas production kinetics

Ahmed EL-WAZIRY, Ahmed AL-HAIDARY, Aly OKAB, Emad SAMARA, Khalid ABDOUN

Effect of freezing rate on goat sperm morphology and DNA integrity

Burcu ÜSTÜNER, Zekariya NUR, Selim ALÇAY, Mehmet Berk TOKER, Hakan SAĞIRKAYA, Mustafa Kemal SOYLU

Incidence and occurrence time of clinical mastitis in Holstein cows

Ismail BOUJENANE, Jalila EL AIMANI, Khalid BY

Effects of alpha lipoic acid and vitamin C supplementation on transforminggrowth factor alpha (TGF α) and transforming growth factor beta 2 (TGF β2)secretion in testicular tissue of diabetic rats

Hamit USLU, Buket BAKIR, Musa KARAMAN, Ebru SARI KARADAĞ, Sevda YILDIZ ELİŞ

Effects of dystocia on milk production and reproduction in subsequent lactation in a Turkish Holstein herd

İbrahim KAYA, Can UZMAY, Tarık AYYILMAZ

Single nucleotide polymorphism identification and characterization of GnRHR gene in Bos taurus and Bos indicus cattle

Ramesha KEREKOPPA, Marappa BASAVARAJU, Akhila RAO, Gujjar Ramachandra GEETHA, Kataktalware MUKUND, Jeyakumar SAKTHIVEL

Effect of organic zinc supplementation on growth, nutrient utilization, and plasma zinc status in lambs

Mokhtar MALLAKI, Mohammad Ali NOROUZIAN, Ali Akbar KHADEM

Alev Gürol BAYRAKTAROĞLU, Özkan ŞİMŞEK, Aytül KÜRÜM, Şevket ARIKAN, Emel ERGÜN

Histological and histochemical studies on the structure of pancreatic Histological and histochemical studies on the structure of pancreatic ducts of the goose (Anser anser)

Turgay DEPREM, Serap Koral TAŞÇI, Seyit Ali BİNGÖL, Ebru Karadağ SARI, Şahin ASLAN, Serap İLHAN

Relationships between sarcomere length and basic composition of infraspinatus and longissimus dorsi muscle

Dominika GUZEK, Dominika GLABSKA, Krzysztof GLABSKI, Grzegorz POGORZELSKI, Jerzy BARSZCZEWSKI, Agnieszka WIERZBICKA