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Effects of osmotic pressure on motility, plasma membrane integrity and viability in fresh and frozen-thawed buffalo spermatozoa

Published online by Cambridge University Press:  01 April 2008

M. I. R. Khan  and
A. Ijaz
M. I. R. Khan
Affiliation:
Department of Theriogenology, Faculty of Veterinary Science, University of Veterinary and Animal Sciences, Lahore 54000, Pakistan
A. Ijaz*
Affiliation:
Department of Theriogenology, Faculty of Veterinary Science, University of Veterinary and Animal Sciences, Lahore 54000, Pakistan
*
E-mail: drijaz80@hotmail.com
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Abstract

In the first experiment, osmotic pressure of semen and seminal plasma in a semen sample from each of the 20 mature Nili-Ravi buffalo bulls was determined. In the second experiment, effects of osmotic pressure on motility (%), plasma membrane integrity (%) and viability (%) in fresh and frozen-thawed semen samples from each of the seven mature Nili-Ravi buffalo bulls was determined. In the first experiment, seminal plasma was harvested by centrifuging semen at 400 × g for 10 min at 37°C and osmotic pressure was determined using an osmometer. In the second experiment, motility (%) was assessed in fresh and frozen-thawed (37°C for 30 s) semen samples using a phase-contrast microscope (×400). Plasma membrane integrity (%) was determined by mixing 50 μl each of fresh and frozen-thawed semen with 500 μl of solution having an osmotic pressure of 50, 100, 150, 190 or 250 mOsm/l (hypotonic treatments of fructose + sodium citrate) and incubating at 37°C for 1 h. Viability (%) of fresh and frozen-thawed spermatozoa before and after challenging them to osmotic pressure (hypotonic treatments) was assessed using supravital stain under a phase-contrast microscope (×400). In the first experiment, the mean ± s.e. osmotic pressures of the buffalo semen and seminal plasma were 268.8 ± 1.17 and 256.0 ± 1.53 mOsm/l, respectively. In the second experiment, motility (%) decreased (P < 0.05) in frozen-thawed semen samples as compared with fresh semen (60.1 ± 1.34 v. 81 ± 1.57, respectively). The plasma membrane integrity (%) and magnitude of osmotic stress in fresh and frozen-thawed semen samples was higher (P < 0.05) at 50, 100, 150 and 190 mOsm/l as compared with 250 mOsm/l. Loss of viability (%) in fresh and frozen-thawed semen samples was higher (P < 0.05) at 50 mOsm/l (59% in fresh, 70% frozen thawed) as compared with other osmotic pressures, while it was lowest at 250 mOsm/l (4.1% for fresh, 9.7% frozen thawed). In conclusion, osmotic pressure of Nili-Ravi buffalo semen and seminal plasma is determined. Furthermore, variation in osmotic pressure below 250 mOsm/l is not favorable to fresh and frozen-thawed buffalo spermatozoa.

Keywords

buffalo semenfresh and frozen-thawed semenosmotic pressureplasma membrane integrityviabilitymotility

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Type
Full Paper
Information
animal , Volume 2 , Issue 4 , April 2008 , pp. 548 - 553
Copyright
Copyright © The Animal Consortium 2008

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References

Ahmad, K, Chaudhry, RA 1980. Cryopreservation of buffalo semen. Veterinary Record 106, 199–201.CrossRefGoogle ScholarPubMed
Blach, EL, Amann, RP, Bowen, RA, Franz, D 1989. Change in quality of stallion spermatozoa during cryopreservation: plasma membrane integrity and motion characteristics. Theriogenology 31, 283–298.CrossRefGoogle ScholarPubMed
Bloom, E 1950. A rapid staining method using eosin-nigrosin to distinguish between live and dead spermatozoa. Animal Breeding Abstracts 18, 1390.Google Scholar
Correa, JR, Zavos, PM 1994. The hypo osmotic swelling test: Its employment as an assay to evaluate the functional integrity of the frozenthawed bovine sperm membrane. Theriogenology 42, 351–360.CrossRefGoogle Scholar
Drevius, LO, Erikson, H 1966. Osmotic swelling of mammalian spermatozoa. Experimental Cell Research 42, 136–156.CrossRefGoogle ScholarPubMed
Drevius, LO 1972. Bull spermatozoa as osmometers. Journal of Reproduction and Fertility 28, 29–39.CrossRefGoogle ScholarPubMed
Hammerstedt, RH, Graham, JK, Nolan, JP 1990. Cryopreservation of mammalian spermatozoa: what we ask them to survive. Journal of Andrology 11, 73–88.CrossRefGoogle ScholarPubMed
Hancock, JL 1951. A staining technique for the study of temperature shock in semen. Nature 167, 323–326.CrossRefGoogle Scholar
Jeyendran, RS, Van Der Van, HH, Perez-Pelaez, M, Crabo, BG, Zaneveld, JL 1984. Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics. Journal of Reproduction and Fertility 70, 219–228.CrossRefGoogle ScholarPubMed
Mandal, DK, Nagpaul, PK, Gupta, AK 2003. Motion characteristics of Murrah buffalo bull spermatozoa in various seasons and its relationship with functional integrity of plasmalemma. Theriogenology 60, 349–358.CrossRefGoogle Scholar
Meyers, SA 2005. Spermatozoa response to osmotic stress. Animal Reproduction Science 89, 57–64.CrossRefGoogle ScholarPubMed
Nie, GJ, Wenzel, GW 2001. Adaptation of the hypoosmotic swelling test to assess functional integrity of stallion spermatozoal plasma membranes. Theriogenology 55, 1005–1018.CrossRefGoogle ScholarPubMed
Petrunkina, AM, Petzoldt, R, Stahlberg, S, Pfeilsticker, J, Beyerbach, M, Bader, H, Töpfer-Petersen, E 2001. Sperm-cell volumetric measurements as parameters in bull semen function evaluation: correlation with nonreturn rate. Andrologia 33, 360–367.CrossRefGoogle ScholarPubMed
Pratap, N, Vishwanatha Reddy, NV, Sarma, PA, Honnappa, TG 2000. Employment of hypotonic swelling test (HOST) to evaluate sperm membrane integrity of fresh and frozen buffalo (Bubalus bubalus) bull semen. Buffalo Journal 2, 207–213.Google Scholar
Quinn, GP, Keough, MJ 2002. Experimental design and data analysis for biologists. Cambridge University Press, Cambridge (DOI: 10.2277/0521009766).CrossRefGoogle Scholar
Rao, PS, Skies, JD, Merilan, CP 1966. Effect of freezing point depression in freezing and storage of bovine semen. Journal of Dairy Science 49, 726–732.Google Scholar
Rasul, Z, Ahmad, N, Anzar, M 2001. Changes in motion characteristics, plasma membrane integrity and acrosome morphology during cryopreservation of buffalo spermatozoa. Journal of Andrology 22, 278–283.CrossRefGoogle ScholarPubMed
Vale, WG 1994. Collection, processing and deep freezing of buffalo semen of buffalo semen. Buffalo Journal 2, 65–72.Google Scholar
Yanagimachi, R 1994. Mammalian fertilization. In The physiology of reproduction (ed. E Knobil and JD Neill), pp. 189–317. Raven Press, New York.Google Scholar