Qualitative ultrastructural changes and morphometry of deccani sheep spermatozoa preserved with egg yolk citrate extender
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Abstract
The present investigation was aimed to study the sequential changes in the sperm cell deterioration dur-ing liquid storage of Deccani sheep breed semen from dilution to 48 h of storage along with its seminal characteris-tics and sperm morphometric measurement. Thus the two Deccani adult rams (aged 2 years), were selected (six ejaculates/each ram) and the collected semen was diluted with Egg yolk citrate extender (EYC) (final concentration - 400 million spermatozoa/0.2 ml semen).Seminal characteristics were assessed along with sperm morphologi-cal changes by Electron microscopy immediately after dilution, at 24 and 48 h of storage, respectively. Sperm morphometry was analysed by Image analysis.The percentage of Individual motility, Live spermatozoa, Acrosomal integrity and Hos-test reactive sperm decreased significantly (P<0.05) from 80.41 to 49.16%, 82.75 to 51.25%, 94.16 to 83% and 76 to 48.58%, respectively during liquid storage of semen from initial dilution to 48 h of storage. The sperm head length (?m), Head width, sperm head area (?m2), sperm head perimeter (?m), mid piece length (?m), proximal mid piece width (?m), distal midpiece width (?m), volume of mid piece (?m3) and acrosomal cap length (?m) were 7.80, 4.33, 26.84, 20.63, 14.03, 0.74, 0.51, 4.54 and 5.24, respectively. Electron microscopic qualitative evaluation revealed that the main site of injury is the apical ridge of ram spermatozoa when stored at 5ºC. The electron density of the mitochondria reduced indicating concomittant depletion of ATP and loss of motility resulting in reduction of fertility.
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Keywords
Liquid preservation, Sperm cell deterioration, Sperm morphometry, Electron microscopy, Mitochondria
References
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Aitken, R.J. and Fisher,H. (1994). Reactive oxygen species generation and human spermatozoa: the balance of benefit and risk. Bioessays. 16:259-267
Alcay, S., Toker, B., Ustuner, B., Nur, Z., Sagirkaya, H. and Soylu, M. K.(2014). Investigation of Relationships between DNA Integrity and Fresh Semen Parameters in Rams. KafkasUniv Vet FakDerg. 20 (5): 793-798
Arthur, G.H., Noakes, D.E. and Pearson, H. (1982). Veterinary Reproduction and Obstetrics. 6thEd. The English Language Book Society and Balliere Tindall, London. pp 517-519
Asadpour, R (2012). Relationship between mineral composition of seminal plasma and semen quality in various ram breeds. Acta Scientiae Veterinariae. 40 (2): 1027
Bharti, M.K., Sinha, M.P., Singh, Balraj and Mahto, D. (2009). Study of semen characteristics of chottanagpuri rams. Indian J. Anim. Reprod. 30: 42-45
Boediono, A., Herdis and Rizal, M. (2004). Preservation of garut rams spermatozoon as a source of male germ plasm. Biotropia. 23: 40 – 46
Boshoff. (2014). The influence of genotype on sperm motility andsperm head morphometry of Merino (Ovisaries) sheep. M. Sc. Thesis (unpublished). Faculty of Agrisciences at Stellenbosch University
Boonkusol, D., Saikhun, K. and Ratanaphumma, P. (2010). Effect of extender and storage time on motility and ultrastructure of cooled-preserved boar spermatozoa. Kasetsart J. (Natural Science) 44:582-589
Campbell, R.C., Dott, H.M. and Glover, T.D. (1956). Nigrosin eosin as stain for differentiating live and dead spermatozoa. J. Agric. Sci. 48, 1–8
Courot, M. and Ortavant, R. (1981). Endocrine control of spermatogenesis in the ram. J Reprod Fertil Suppl., 30:47– 60
David, I. (2008). Analyse génétique et modélisation de la production de semence et de la réussite de l’insémination artificielle en ovin Life sciences. AgroParisTech, Paris, pp. 209
Cseh. S., Faigl, V., Vass, N., Javor, A., Kulcsar, M., Solti, L. and Amiridis, G.(2012). Artificial insemination of small ruminants — A Review. Acta VeterinariaHungarica. 60 (1): 115–129
Elsharif, B.A.A. (2010). Reproductive traits and freezability of semen of rams from two desert sheep ecotypes in the Sudan. Department of Reproduction and Obstetrics, Ph.D. thesis (unpublished). Faculty of Veterinary Medicine. University of Khartoum. Sudan
Fiser, P. S. and Fairful, R. W. (1989). The effect of glycerol related osmotic changes on post thaw motility and acrosomal integrity of ram spermatozoa. Cryobiology. 26:64-69
Gundogan, M., 2007. Seasonal variation in serum testosterone, T3 and andrological parameters of two Turkish sheep breeds. Small Rumin Res. 67: 312-316
Herman, H. A. and Madden, F.W. (1953). The Artificial Insemination of Dairy Cattle. A hand book and laboratory manual. Lucase Brothers Publishers, Colombia, Missori, USA
Herman, H.A., Mitchell, J.R. and Doak, G.A. (1994). The Artificial Insemination and Embryo Transfer of Dairy and Beef Cattle. Danville, IL: Interstate Publishers, Inc. Danville, Illinois, USA
Jeyendran, R.S., Van der Ven, H.H. and Perez-Pelaez, M. (1984). Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics. J. Reprod. Fertil. 70:219-228
Jones R.C. (1973). The effects of dilution, egg yolk and cooling to 5ºC on the ultrastructure of ram spermatozoa. J. Reprod. Fert. 35: 311-320
Kakar. S.S. and Anand, S R. (1984). Acrosomal Damage and Enzyme Leakage during Freeze Preservation of Buffalo Spermatozoa. Indian Journal of Experimental Biology. 22: 5-10
Kayali, I.M., Rashed, M.A., Anous, M.R., Sallam, M.A.A. and Atta, A.H. (2014). Physiological and molecular characterization of some Egyptian sheep breeds.J. Biol. Chem. Environ. Sci., 9 (1): 121-140
Lamirande, E., Jiang, H., Zini, A. and Kodama, C. (1997). Gagnon Reactive oxygen species and sperm physiology. Reviews of Reproduction. 2(1):48–54
Marai, I.F.M. and Owen. J.B. (2013). New Techniques in Sheep Production. Butterworth-Heinemann publishers. pp 99
Moghaddam, G.H., Pourseif, M.M. and Rafat, S.A. (2012). Seasonal variation in semen quantity and quality traits of Iranian crossbred rams.Slovak J. Anim. Sci., 45 (3): 67-75
Mortimer ,S.T (2000). CASA-Practical aspects. J. Androl., 21: 515-524
Nalley, W.M.M. and Arifiantini, R.I. (2013). The hypo-osmotic swelling test in fresh garut ram spermatozoa. J.Indonesian Trop. Anim.Agric. 38(4): 212-216
Pesch, S. and Bergmann, M. (2006). Structure of mammalian spermatozoa in respect to viability, fertility and cryopreservation. Micron. 37: 597-612
Prinosilova, P., Sedlackova, M., Kopecka, V. and Hlavicova, J. (2012). Boar sperm head membrane damage during cryopreservation evaluated by electron microscopy. Research in pig breeding. 6 (2): 58-61
Rao, A.R. (1971). Changes in the morphology of sperm during their passage through the genital tract in bulls with normal and impaired spermatogenesis. Ph.D. Thesis (unpublished). Roy. Vet. Coll. Stockholm, Sweden
Ros-Santaella, J.L., Dom?nguez-Rebolledo, A.E. and Garde, J.J.(2014). Sperm Flagellum Volume Determines Freezability in RedDeer Spermatozoa. Plos one. 9 (11):1-11
Salamon, S. and Maxwell, M.C. (2000): Storage of ram semen. Anim. Reprod. Sci. 62:77–111
Saxena, M.S. (2000). Veterinary andrology and Artificial Insemination. 1st Ed. CBS publishers and Distributors. New Delhi (India)
Shimizu, T. and Johnson, K.A. (1983). Kinetic evidence for multiple dynein ATPase sites. The Journal of Biological Chemistry, 258(22): 3841–3846
Shukla, M.K. (2011). Applied veterinary andrology and frozen semen technology. New delhi publishing agency. New Delhi (India)
Toe, F., Lahlou-Kassi, A. and Mukasa-Mugerwa, E. (1994). Semen characteristics of Ile-de-France rams of different age and physical condition. Theriogenology. 42(2):
321-326
Watson, P.F. (1975). Use of Giemsa stain to detect changes in the acrosome of frozen ram spermatozoa. Vet. Record. 97:12-15
Yaniz, J.L., Santolaria, P., Marco-Aguado, M.A and Lopez-Gatius, F. (2008). Use of image analysis to assess the plasma membrane integrity of ram spermatozoa in different diluents. Theriogenology. 70:192–198
Yaniz, J.L., Vicente-Fiel, S., Capistros, S., Palacín, I. and Santolaria, P. (2012). Automatic evaluation of ram sperm Morphometry.Theriogenology. 77 :
1343–1350
Yotov, S., Fasulkov, I. and Vassilev, N. (2011). Effect of ejaculation frequency on spermatozoa survival in diluted semen from Pleven Blackhead rams. Turk. J. Vet. Anim. Sci. 35(2): 117-122
Youngquist. (2007). Current therapy in large animal theriogenology. 2nd Ed. Saunders Elsevier. Philadelphia, USA: pp.620-641
Aitken, R.J. and Fisher,H. (1994). Reactive oxygen species generation and human spermatozoa: the balance of benefit and risk. Bioessays. 16:259-267
Alcay, S., Toker, B., Ustuner, B., Nur, Z., Sagirkaya, H. and Soylu, M. K.(2014). Investigation of Relationships between DNA Integrity and Fresh Semen Parameters in Rams. KafkasUniv Vet FakDerg. 20 (5): 793-798
Arthur, G.H., Noakes, D.E. and Pearson, H. (1982). Veterinary Reproduction and Obstetrics. 6thEd. The English Language Book Society and Balliere Tindall, London. pp 517-519
Asadpour, R (2012). Relationship between mineral composition of seminal plasma and semen quality in various ram breeds. Acta Scientiae Veterinariae. 40 (2): 1027
Bharti, M.K., Sinha, M.P., Singh, Balraj and Mahto, D. (2009). Study of semen characteristics of chottanagpuri rams. Indian J. Anim. Reprod. 30: 42-45
Boediono, A., Herdis and Rizal, M. (2004). Preservation of garut rams spermatozoon as a source of male germ plasm. Biotropia. 23: 40 – 46
Boshoff. (2014). The influence of genotype on sperm motility andsperm head morphometry of Merino (Ovisaries) sheep. M. Sc. Thesis (unpublished). Faculty of Agrisciences at Stellenbosch University
Boonkusol, D., Saikhun, K. and Ratanaphumma, P. (2010). Effect of extender and storage time on motility and ultrastructure of cooled-preserved boar spermatozoa. Kasetsart J. (Natural Science) 44:582-589
Campbell, R.C., Dott, H.M. and Glover, T.D. (1956). Nigrosin eosin as stain for differentiating live and dead spermatozoa. J. Agric. Sci. 48, 1–8
Courot, M. and Ortavant, R. (1981). Endocrine control of spermatogenesis in the ram. J Reprod Fertil Suppl., 30:47– 60
David, I. (2008). Analyse génétique et modélisation de la production de semence et de la réussite de l’insémination artificielle en ovin Life sciences. AgroParisTech, Paris, pp. 209
Cseh. S., Faigl, V., Vass, N., Javor, A., Kulcsar, M., Solti, L. and Amiridis, G.(2012). Artificial insemination of small ruminants — A Review. Acta VeterinariaHungarica. 60 (1): 115–129
Elsharif, B.A.A. (2010). Reproductive traits and freezability of semen of rams from two desert sheep ecotypes in the Sudan. Department of Reproduction and Obstetrics, Ph.D. thesis (unpublished). Faculty of Veterinary Medicine. University of Khartoum. Sudan
Fiser, P. S. and Fairful, R. W. (1989). The effect of glycerol related osmotic changes on post thaw motility and acrosomal integrity of ram spermatozoa. Cryobiology. 26:64-69
Gundogan, M., 2007. Seasonal variation in serum testosterone, T3 and andrological parameters of two Turkish sheep breeds. Small Rumin Res. 67: 312-316
Herman, H. A. and Madden, F.W. (1953). The Artificial Insemination of Dairy Cattle. A hand book and laboratory manual. Lucase Brothers Publishers, Colombia, Missori, USA
Herman, H.A., Mitchell, J.R. and Doak, G.A. (1994). The Artificial Insemination and Embryo Transfer of Dairy and Beef Cattle. Danville, IL: Interstate Publishers, Inc. Danville, Illinois, USA
Jeyendran, R.S., Van der Ven, H.H. and Perez-Pelaez, M. (1984). Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics. J. Reprod. Fertil. 70:219-228
Jones R.C. (1973). The effects of dilution, egg yolk and cooling to 5ºC on the ultrastructure of ram spermatozoa. J. Reprod. Fert. 35: 311-320
Kakar. S.S. and Anand, S R. (1984). Acrosomal Damage and Enzyme Leakage during Freeze Preservation of Buffalo Spermatozoa. Indian Journal of Experimental Biology. 22: 5-10
Kayali, I.M., Rashed, M.A., Anous, M.R., Sallam, M.A.A. and Atta, A.H. (2014). Physiological and molecular characterization of some Egyptian sheep breeds.J. Biol. Chem. Environ. Sci., 9 (1): 121-140
Lamirande, E., Jiang, H., Zini, A. and Kodama, C. (1997). Gagnon Reactive oxygen species and sperm physiology. Reviews of Reproduction. 2(1):48–54
Marai, I.F.M. and Owen. J.B. (2013). New Techniques in Sheep Production. Butterworth-Heinemann publishers. pp 99
Moghaddam, G.H., Pourseif, M.M. and Rafat, S.A. (2012). Seasonal variation in semen quantity and quality traits of Iranian crossbred rams.Slovak J. Anim. Sci., 45 (3): 67-75
Mortimer ,S.T (2000). CASA-Practical aspects. J. Androl., 21: 515-524
Nalley, W.M.M. and Arifiantini, R.I. (2013). The hypo-osmotic swelling test in fresh garut ram spermatozoa. J.Indonesian Trop. Anim.Agric. 38(4): 212-216
Pesch, S. and Bergmann, M. (2006). Structure of mammalian spermatozoa in respect to viability, fertility and cryopreservation. Micron. 37: 597-612
Prinosilova, P., Sedlackova, M., Kopecka, V. and Hlavicova, J. (2012). Boar sperm head membrane damage during cryopreservation evaluated by electron microscopy. Research in pig breeding. 6 (2): 58-61
Rao, A.R. (1971). Changes in the morphology of sperm during their passage through the genital tract in bulls with normal and impaired spermatogenesis. Ph.D. Thesis (unpublished). Roy. Vet. Coll. Stockholm, Sweden
Ros-Santaella, J.L., Dom?nguez-Rebolledo, A.E. and Garde, J.J.(2014). Sperm Flagellum Volume Determines Freezability in RedDeer Spermatozoa. Plos one. 9 (11):1-11
Salamon, S. and Maxwell, M.C. (2000): Storage of ram semen. Anim. Reprod. Sci. 62:77–111
Saxena, M.S. (2000). Veterinary andrology and Artificial Insemination. 1st Ed. CBS publishers and Distributors. New Delhi (India)
Shimizu, T. and Johnson, K.A. (1983). Kinetic evidence for multiple dynein ATPase sites. The Journal of Biological Chemistry, 258(22): 3841–3846
Shukla, M.K. (2011). Applied veterinary andrology and frozen semen technology. New delhi publishing agency. New Delhi (India)
Toe, F., Lahlou-Kassi, A. and Mukasa-Mugerwa, E. (1994). Semen characteristics of Ile-de-France rams of different age and physical condition. Theriogenology. 42(2):
321-326
Watson, P.F. (1975). Use of Giemsa stain to detect changes in the acrosome of frozen ram spermatozoa. Vet. Record. 97:12-15
Yaniz, J.L., Santolaria, P., Marco-Aguado, M.A and Lopez-Gatius, F. (2008). Use of image analysis to assess the plasma membrane integrity of ram spermatozoa in different diluents. Theriogenology. 70:192–198
Yaniz, J.L., Vicente-Fiel, S., Capistros, S., Palacín, I. and Santolaria, P. (2012). Automatic evaluation of ram sperm Morphometry.Theriogenology. 77 :
1343–1350
Yotov, S., Fasulkov, I. and Vassilev, N. (2011). Effect of ejaculation frequency on spermatozoa survival in diluted semen from Pleven Blackhead rams. Turk. J. Vet. Anim. Sci. 35(2): 117-122
Youngquist. (2007). Current therapy in large animal theriogenology. 2nd Ed. Saunders Elsevier. Philadelphia, USA: pp.620-641
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Qualitative ultrastructural changes and morphometry of deccani sheep spermatozoa preserved with egg yolk citrate extender. (2017). Journal of Applied and Natural Science, 9(3), 1515-1521. https://doi.org/10.31018/jans.v9i3.1394
