The aim of this study was to compare the effect of different packaging systems on some parameters of cryopreserved canine spermatozoa. The experimental material consisted of the sperm-rich fractions of ejaculates collected from four Beagle dogs. Semen samples for cryopreservation were stored in 0.25 ml plastic straws and two aluminum tubes with a total volume of 5.0 ml. Semen was frozen in static nitrogen vapor for 10 minutes (0.25 ml straws) or 15 and 20 minutes (aluminum tubes). Post-thaw assessments involved the determination of sperm motility parameters using a computer assisted sperm analyzer (CASA), sperm plasma membrane integrity (SPMI), mitochondrial membrane potential (MMP) and acrosome integrity (normal apical ridge, NAR). Regardless of the packaging system applied, no significant differences in total sperm motility (TMOT) or selected kinematic parameters were observed after freezing-thawing. However, spermatozoa frozen in 0.25 mL straws were characterized by improved functionality, in particular mitochondrial function, after thawing. The results indicate that large quantities of canine semen can be frozen in aluminum tubes. Further studies are required, however, to evaluate different freezing and thawing rates of aluminum tubes.
Ali SB, Tischner M (1988) Freezing of ram semen in aluminium packets and deep cervical insemination of ewes with a modified pipette. Med Weter 44: 294-297.
Anderson K (1972) Fertility of frozen dog semen. Acta Vet Scand 13: 128-130.
Bielas W, Dubiel A, Rząsa A, Twardoń J, NiżańskiW (2010) Effect of packaging form and methods of boar semen freezing on chosen reproductive indexes of sows. Med Weter 66: 711-715.
Bielas W, Dubiel A, Niżański W (2003) Effects of cryopreservation methods, packaging systems and the thermoresistance test on the post-thaw quality of boar semen. Med Weter 59: 172-175.
Ericsson BM, Petersson H, Rodriguez-Martinez H (2002) Field fertility with exported boar semen frozen in the new flatpack container. Theriogenology 58: 1065-1079.
Farstad WK (2010) Artificial insemination in dogs. In: England G, von Heimendahl A (eds) BSAVA Manual of canine and feline reproduction and neonatology, 2nd ed., British Small Animal Veterinary Association, Quedgeley, Gloucester, pp 80-88.
Fraser L, Strzeżek J (2007) Effect of different procedures of ejaculate collection, extenders and packages on DNA integrity of boar spermatozoa following freezing-thawing. Anim Reprod Sci 99: 317-329.
Garner DL, Johnson LA (1995) Viability assessment of mammalian sperm using SYBR-14 and propidium iodide. Biol Reprod 53: 276-284.
Ivanova-Kicheva MG, Bobadov N, Somlev B (1997) Cryopreservation of canine semen in pellets and in 5-mL aluminum tubes using three extenders. Theriogenology 48: 1343-1349.
Kosiniak K, Bittmar A, Sallwerk K (1992) Relationship between biochemical components of dog seminal plasma and semen freezability. Proceedings of 12th International Congress on Animal Reproduction, Hague, pp 1791-1792.
Martins-Bessa A, Rocha A, Mayenco-Aguirre A (2006) Comparing ethylene glycol with glycerol for cryopreservation of canine semen in egg-yolk TRIS extenders. Theriogenology 66: 2047-2055.
Niżański W (2006) Intravaginal insemination of bitches with fresh and frozen-thawed semen with addition of prostatic fluid: use of an infusion pipette and the Osiris catheter. Theriogenology 66: 470-483.
Niżański W, Dubiel A (2003) The effect of packaging systems on post-thaw spermatozoal characteristics in cryopreserved dog semen. Med Weter 59: 829-833.
Nothling JO, Shuttleworth R (2005) The effect of straw size, freezing rate and thawing rate upon post-thaw quality of dog semen. Theriogenology 63: 1469-1480.
Peńa AI, Barrio F, Quintela LA, Herradón PG (1998) Effect of different glycerol treatments on frozen-thawed dog sperm longevity and acrosomal integrity. Theriogenology 50: 163-174.
Peña FJ, Núñez-Martínez I, Morán JM (2006) Semen technologies in dog breeding: an update. Reprod Domest Anim 41, (Suppl 2): 21-29.
Rota A, Ström B, Linde-Forsberg C (1995) Effects of seminal plasma and three extenders on canine semen stored at 4 degrees C. Theriogenology 44: 885-900.
Rota A, Linde-Forsberg C, Vannozzi J, Romagnoli S, Rodriguez- Martinez H (1998) Cryosurvival of dog spermatozoa at different glycerol concentrations and freezing/thawing rates. Reprod Domest Anim 33: 355-361.
Silva AR, Cardoso RC, Silva LD (2006) Influence of temperature during glycerol addition and post-thaw dilution on the quality of canine frozen semen. Reprod Domest Anim 41: 74-78.
Schäfer-Somi S, Kluger S, Knapp E, Klein D, Aurich C (2006) Effects of semen extender and semen processing on motility and viability of frozen-thawed dog spermatozoa. Theriogenology 66: 173-182.
Strzeżek J, Glogowski J, Hopfer E, Wojtkiewicz K (1985) The „Kortowska” method of a boar’s semen freezing. Med Weter 41: 349-353.
Tischner M (1979) Evaluation of deep-frozen semen in stallions. J Reprod Fertil Suppl 27: 53-59.
Thomas CA, Garner DL, DeJarnette JM, Marshall CE (1998) Effect of cryopreservation of bovine sperm organelle function and viability as determined by flow cytometry. Biol Reprod 58: 786-793.
Thomas PG, Larsen RE, Burns JM, Hahn CN (1993) A comparison of three packaging techniques using two extenders for the cryopreservation of canine semen. Theriogenology 40: 1199-1205.
Watson PF (1975) Use of a Giemsa stain to detect changes in acrosomes of frozen ram spermatozoa. Vet Rec 97: 12-15.