Öz
The aim of this study was to determine the influence of different cooling rates and trehalose concentrations on standard semen parameters after the freezing-thawing of ram spermatozoa. Ejaculates collected using artificial vagina from 4 Akkaraman rams were evaluated and pooled at room temperature. Semen samples diluted with a Tris-based extender containing different concentrations (50, 100 and 150 mM) of trehalose and no additives (control) were cooled to 5°C and frozen at slow (a rate of 0.5°C/min from 5°C to –25°C; -50°C/min from –25°C to -130°C), and fast (a rate of 10°C/min from 5°C to -25°C; -50°C/min from -25°C to -130°C) cooling rates and plunged into liquid nitrogen. Frozen straws were thawed individually at 37°C for 20 sec in a water bath for evaluation. In conclusion, 50 and 100 mM trehalose and slow cooling rate were found to significantly (p<0.05) improve the freezability of ram spermatozoa which were assessed regarding post-thawing sperm motility, morphologic sperm abnormalities, viability and membrane integrity. But, 100 mM trehalose concentration and slow cooling rate were observed superior than other treatment groups
Anahtar Kelimeler
Fast freezing ram semen semen parameters slow freezing trehalose
Kaynakça
- Aboagla EME, TeradaT (2003): Trehalose-enhanced fluidity of the goat sperm membrane and its protection during freezing. Biol Reprod, 69, 1245-1250.
- Aisen EG, Medina VH, Venturino A (2002): Cryopreservation and post-thawed fertility of ram semen frozen in different trehalose concentrations. Theriogenology, 57, 1801-1088.
- Anel L, Boixo JC, Anel E, Carbajo M, Domínguez JC, Olmedo JA, Melcon C (1992): Fertility of Churra Ewes following intrauterine insemination by laparoscopy with frozen-thawed semen. In: Proceedings of the 12th International Congress on Animal Reproduction and AI. The Hage, Holland,1384–1386 pp.
- Anel L, Alvarez M, Martinez-Pastor F, Garcia-Macias V, Anel E, de Paz P (2006): Improvement strategies in ovine artificial insemination. Reprod Dom Anim, 41, 30- 42.
- Bakas LS, Disalvo EA (1991): Effect of Ca2+ on the cryoprotective action of trehalose. Cryobiology, 28, 347- 353.
- Bucak MN, Ateşşahin A, Varışlı Ö, Yüce A, Tekin N, Akçay A (2007): The influence of trehalose, taurine, cysteamine and hyaluronanon ram semen: Microscobic and oxidative parameters after freze-thawing process. Theriogenology, 67, 1060-1067.
- Buckett WM, Luckas MJ, Aird IA, Farquharson RG, Kingsland CR, Lewis-Jones DI (1997): The hypo-osmotic swelling test in recurrent miscarriage. Fertil and Steril, 68, 506-509.
- Byrne GP, Lonergan P, Wade M, Dufy P, Donovan A, Hanrahan JP, Boland M P (2000): Effect of freezing rate of ram spermatozoa on subsequent fertility in vivo and in vitro. Anim Reprod Sci, 62, 265-275.
- Chen Y, Foote RH, Brockett CC (1993): Effect of sucrose, trehalose, hypotaurine, taurine, and blood serum on survival of frozen bull sperm. Cryobiology, 30, 423-431.
- Curry MR, Millar JD, Watson PF (1994): Calculated optimal cooling rates for ram and human sperm cryopreservation fail to conform with empirical observations. Biol Reprod, 51, 1014-1021.
- Daniel WW (1991): Analysis of variance. 274-320. In: WW Daniel (Ed), Biostatistic: A Foundation for Analysis in the Health Sciences. John Wiley& Sons, Hoboken.
- Evans G, Maxwell WMC (1987): Handling and examination semen. 93-106. In: WMC Maxwell (Ed), Salamon’s Artificial Insemination of Sheep and Goat. Butterworths, Sidney.
- Gil J, Lundeheim N, Söderquist L, Rodriguez-Martinez H.(2003): Influence of extender, temperature, and addition of glycerol on post-thaw sperm parameters in ram semen. Theriogenology, 59,1241-1255.
- Henry MA, Noiles EE, Gao D, Mazur P, Critser JK (1993): Cryopreservation of human spermatozoa. IV. The effects of cooling rate and warming rate on the maintenance of motility plasma membrane integrity and mithocondrial function. Fertil and Steril, 60, 911-918.
- Kumar S, Millar JD, Watson PF (2003): The effect of cooling rate on the survival of cryopreserved bull, ram, and boar spermatozoa: a comparison of two controlled- rate cooling machines. Cryobiology, 46, 246–253.
- Liu Z, Foote RH, Brockett CC (1998): Survival of bull sperm frozen at different rates in media varying in osmolarity. Cryobiology, 37, 219-230.
- Marti JI, Marti E, Cebrian-Perez JA, Muino-Blanco T (2003): Survival rate and antioxidant enzyme activity of ram spermatozoa after dilution with different extenders or selection by a dextran swim-up procedure. Theriogenology, 60, 1025-1037.
- Matsuoka T, Imai H, Kohno H, Fukui Y (2006): Effects of bovine serum albumine and trehalose in semen diluents for improvement of frozen-thawed ram spermatozoa. J Reprod Dev, 52, 675-683.
- Mazur P (1965): Causes of injury in frozen and thawed cells. Fed. Proc. 24, 175-182.
- Molinia FC, Evans G, Maxwell WMC (1994): Effects of monosaccharides and disaccharides in tris-based diluents on motility, acrosome integrity and fertility of pellet frozen ram spermatozoa. Anim Reprod Sci, 36,113-122. 21. O’Neill
- DJ (1998): Studies Related to the Cryopreservation of Ram Spermatozoa Thesis, National University of Ireland, Dublin. Rodriguez-Martinez H, Tienthai P, Suzuki K,
- Funahashi H, Ekwall H, Johannisso A (2001): Involvement of oviduct in sperm capacitacion and oocyte development in pigs. Reproduction, 58, 129-145.
- Rudolph AS, Crowe HJ (1985): Membrane stabilization during freezing: the role of two natural cryoprotectants, trehalose and praline. Cryobiology, 22, 367-377.
- Salamon S, Maxwell WMC (1995): Frozen storage of ram semen. I. Procesing, freezing, thawing and fertility after cervical insemination. Anim Reprod Sci, 37, 185– 249.
- Salamon S, MaxwellWM (2000): Storage of ram semen. Anim Reprod Sci, 62, 77-111.
- Schafer S, Holzmann A. (2000): The use of transmigration and spermac stain to evaluate epididymal cat spermatozoa. Anim Reprod Sci, 59, 201-211.
- Seigneurin F, Blesbois E (1995): Effects of the freezing rate on viability and fertility of frozen-thawed fowl spermatozoa. Theriogenology, 43, 1351–1358.
- Watson PF (1995): Recent developments and concepts in the cryopreservation of spermatozoa and the assessment of their post-thawing function. Reprod Fertil Dev, 7, 871-891.
- Woelders H, Matthijs A, Engel B (1997): Effects of trehalose and sucrose, osmolality of the freezing medium, and cooling rate on viability and intactness of bull sperm after freezing and thawing. Cryobiology, 35, 93-105.
Öz
Bu çalışmanın amacı dondurma/çözdürme sonrası farklı soğutma oranları ve trehaloz konsantrasyonlarının standart spermatolojik parametreler üzerine etkisini saptamaktır. Sun’i vagina kullanılarak 4 Akkaraman koçtan toplanan ejakülatlar oda sıcaklığında değerlendirildi ve karıştırıldı. Farklı konsantrasyonlarda (50, 100 ve 150 mM) trehaloz içeren ve içermeyen (control) tris bazlı solusyonla sulandırılmış sperma örnekleri 5°C’a soğutuldu ve yavaş (dakikada 0.5°C ile 5°C’dan –25°C’a; dakikada -50°C ile –25°C’dan -130°C’a) ve hızlı (dakikada 10°C ile 5°C’dan -25°C’a; dakikada -50°C ile -25°C’dan -130°C’a) soğutma oranlarında donduruldu ve sıvı azota daldırıldı. Dondurulmuş payetler değrlendirme için bireysel olarak su banyosunda 37°C’da 20 saniyede çözdürüldü. Sonuç olarak, çözdürme sonrası spermatozoa motilitesi, morfolojik spermatozoa anomalileri, canlılığı ve membrane bütünlüğü değerlendirilen koç spermatozoasının dondurulabilirliğini, 50 ve 100 mM trehaloz ile yavaş dondurma oranının önemli ölçüde (p<0.05) geliştirdiği bulundu. Fakat, 100 mM trehaloz konsantrasyonu ve yavaş soğutma oranının diğer tedavi gruplarından daha üstün olduğu gözlendi
Anahtar Kelimeler
Hızlı dondurma koç sperması sperma parametreleri yavaş dondurma trehaloz
Kaynakça
- Aboagla EME, TeradaT (2003): Trehalose-enhanced fluidity of the goat sperm membrane and its protection during freezing. Biol Reprod, 69, 1245-1250.
- Aisen EG, Medina VH, Venturino A (2002): Cryopreservation and post-thawed fertility of ram semen frozen in different trehalose concentrations. Theriogenology, 57, 1801-1088.
- Anel L, Boixo JC, Anel E, Carbajo M, Domínguez JC, Olmedo JA, Melcon C (1992): Fertility of Churra Ewes following intrauterine insemination by laparoscopy with frozen-thawed semen. In: Proceedings of the 12th International Congress on Animal Reproduction and AI. The Hage, Holland,1384–1386 pp.
- Anel L, Alvarez M, Martinez-Pastor F, Garcia-Macias V, Anel E, de Paz P (2006): Improvement strategies in ovine artificial insemination. Reprod Dom Anim, 41, 30- 42.
- Bakas LS, Disalvo EA (1991): Effect of Ca2+ on the cryoprotective action of trehalose. Cryobiology, 28, 347- 353.
- Bucak MN, Ateşşahin A, Varışlı Ö, Yüce A, Tekin N, Akçay A (2007): The influence of trehalose, taurine, cysteamine and hyaluronanon ram semen: Microscobic and oxidative parameters after freze-thawing process. Theriogenology, 67, 1060-1067.
- Buckett WM, Luckas MJ, Aird IA, Farquharson RG, Kingsland CR, Lewis-Jones DI (1997): The hypo-osmotic swelling test in recurrent miscarriage. Fertil and Steril, 68, 506-509.
- Byrne GP, Lonergan P, Wade M, Dufy P, Donovan A, Hanrahan JP, Boland M P (2000): Effect of freezing rate of ram spermatozoa on subsequent fertility in vivo and in vitro. Anim Reprod Sci, 62, 265-275.
- Chen Y, Foote RH, Brockett CC (1993): Effect of sucrose, trehalose, hypotaurine, taurine, and blood serum on survival of frozen bull sperm. Cryobiology, 30, 423-431.
- Curry MR, Millar JD, Watson PF (1994): Calculated optimal cooling rates for ram and human sperm cryopreservation fail to conform with empirical observations. Biol Reprod, 51, 1014-1021.
- Daniel WW (1991): Analysis of variance. 274-320. In: WW Daniel (Ed), Biostatistic: A Foundation for Analysis in the Health Sciences. John Wiley& Sons, Hoboken.
- Evans G, Maxwell WMC (1987): Handling and examination semen. 93-106. In: WMC Maxwell (Ed), Salamon’s Artificial Insemination of Sheep and Goat. Butterworths, Sidney.
- Gil J, Lundeheim N, Söderquist L, Rodriguez-Martinez H.(2003): Influence of extender, temperature, and addition of glycerol on post-thaw sperm parameters in ram semen. Theriogenology, 59,1241-1255.
- Henry MA, Noiles EE, Gao D, Mazur P, Critser JK (1993): Cryopreservation of human spermatozoa. IV. The effects of cooling rate and warming rate on the maintenance of motility plasma membrane integrity and mithocondrial function. Fertil and Steril, 60, 911-918.
- Kumar S, Millar JD, Watson PF (2003): The effect of cooling rate on the survival of cryopreserved bull, ram, and boar spermatozoa: a comparison of two controlled- rate cooling machines. Cryobiology, 46, 246–253.
- Liu Z, Foote RH, Brockett CC (1998): Survival of bull sperm frozen at different rates in media varying in osmolarity. Cryobiology, 37, 219-230.
- Marti JI, Marti E, Cebrian-Perez JA, Muino-Blanco T (2003): Survival rate and antioxidant enzyme activity of ram spermatozoa after dilution with different extenders or selection by a dextran swim-up procedure. Theriogenology, 60, 1025-1037.
- Matsuoka T, Imai H, Kohno H, Fukui Y (2006): Effects of bovine serum albumine and trehalose in semen diluents for improvement of frozen-thawed ram spermatozoa. J Reprod Dev, 52, 675-683.
- Mazur P (1965): Causes of injury in frozen and thawed cells. Fed. Proc. 24, 175-182.
- Molinia FC, Evans G, Maxwell WMC (1994): Effects of monosaccharides and disaccharides in tris-based diluents on motility, acrosome integrity and fertility of pellet frozen ram spermatozoa. Anim Reprod Sci, 36,113-122. 21. O’Neill
- DJ (1998): Studies Related to the Cryopreservation of Ram Spermatozoa Thesis, National University of Ireland, Dublin. Rodriguez-Martinez H, Tienthai P, Suzuki K,
- Funahashi H, Ekwall H, Johannisso A (2001): Involvement of oviduct in sperm capacitacion and oocyte development in pigs. Reproduction, 58, 129-145.
- Rudolph AS, Crowe HJ (1985): Membrane stabilization during freezing: the role of two natural cryoprotectants, trehalose and praline. Cryobiology, 22, 367-377.
- Salamon S, Maxwell WMC (1995): Frozen storage of ram semen. I. Procesing, freezing, thawing and fertility after cervical insemination. Anim Reprod Sci, 37, 185– 249.
- Salamon S, MaxwellWM (2000): Storage of ram semen. Anim Reprod Sci, 62, 77-111.
- Schafer S, Holzmann A. (2000): The use of transmigration and spermac stain to evaluate epididymal cat spermatozoa. Anim Reprod Sci, 59, 201-211.
- Seigneurin F, Blesbois E (1995): Effects of the freezing rate on viability and fertility of frozen-thawed fowl spermatozoa. Theriogenology, 43, 1351–1358.
- Watson PF (1995): Recent developments and concepts in the cryopreservation of spermatozoa and the assessment of their post-thawing function. Reprod Fertil Dev, 7, 871-891.
- Woelders H, Matthijs A, Engel B (1997): Effects of trehalose and sucrose, osmolality of the freezing medium, and cooling rate on viability and intactness of bull sperm after freezing and thawing. Cryobiology, 35, 93-105.
Ayrıntılar
| Birincil Dil | İngilizce |
|---|---|
| Konular | Veteriner Cerrahi |
| Diğer ID | JA53FU45MJ |
| Bölüm | Araştırma Makalesi |
| Yazarlar | |
| Yayımlanma Tarihi | 1 Haziran 2009 |
| Yayımlandığı Sayı | Yıl 2009Cilt: 56 Sayı: 2 |