Skip to main content

Advertisement

SpringerLink
Log in

Development of a new method to preserve caprine cauda epididymal spermatozoa in-situ at -10°C with electrolyte free medium

  • Animal Experimentation in Assisted Reproduction
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

In-situ preservation of cauda epididymal spermatozoa at -10°C with electrolyte free media for obtaining maximum functional gametes than preservation at 5°C.

Methods

Electrolyte free media prepared with soybean lecithin-glycerol, Coenzyme Q10 — glycerol and soybean lecithin — Coenzyme Q10— glycerol were inoculated separately into ligated cauda epididymides, equilibrated 2 h at 5°C, wrapped with aluminium foil and freezed at — 10°C. Spermatozoan characters were evaluated 7 and 21 days after thawing at 38.5°C in a water bath for 5 min.

Results

Spermatozoan characteristics were diminished gradually and significantly (p < 0.001, p < 0.05) between the media and observation days. Soybean lecithin-CoenzymeQ10-glycerol effectively protected spermatozoa against cold shock where spermatozoan progressive motility, viability, hypo-osmotic swelling positivity were 30.2 ± 0.62; 45.2 ± 0.82 and 41.6 ± 0.79 percent respectively on day 21.

Conclusion

This method can be adopted in field conditions for transportation of frozen epididymides and re-utilization of maximum functional gametes to conserve valuable animals after postmortem / slaughter.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Foote RH. Letter to the Editor. J. Androl. 2000;21(3):355.

    CAS  PubMed  Google Scholar 

  2. Anel L, Guerra C, Alvarez M, Kaabi M, Anel E, Boixo JC, et al. Basic parameters in spermatozoa recovered post-mortem from the Spanish Cantabrian Chamois (Rupicapra pyrenaica parva). Theriogenology. 2000;53(1):323. abstract.

    Google Scholar 

  3. Blash S, Melican D, Gavin W. Cryopreservation of epididymal sperm obtained at necropsy from goats. Theriogenology. 2000;54(6):899–905.

    Article  CAS  PubMed  Google Scholar 

  4. Kilian I, Lubbe K, Bartels P, Friedmann Y, Denniston RS. Evaluating epididymal sperm of African wild ruminants: Longevity when stored at 4°C and viability following cryopreservation. Theriogenology. 2000;53(1):330. abstract.

    Google Scholar 

  5. Lubbe K, Bartels P, Kilian I, Friedmann Y, Godke RA. Comparing motility and morphology of horse, zebra and rhinoceros epididymal spermatozoa when cryopreserved with two different cryodiluents or stored at 4°C. Theriogenology. 2000;53(1):338. abstract.

    Google Scholar 

  6. Soler AJ, Perez-guzman MD, Grade J. Storage of red deer epididymides for four days at 5°C: effects on motility, viability and morphological integrity. J. Exp. Zool. Part A: Comparative Exp. Biol. 2003;295A:188–99.

    Article  Google Scholar 

  7. Martinez-Pastor F, Guerra C, Kabbi M, Diaz AR, Anel E, Herraez P, et al. Decay of sperm obtained from epididymis of wild ruminants depending on postmortem time. Theriogenology. 2005;63(1):24–40.

    Article  CAS  PubMed  Google Scholar 

  8. Dong Q, Rodenberg SE, Huang C, Vandevoort CA. Cryopreservation of Rhesus monkey (Macaca muitatta) epididymal spermatozoa before and after refrigerated storage. J. Androl. 2008;29(3):283–92.

    Article  PubMed  Google Scholar 

  9. James AN, Green H, Hoffman S, Landry AM, Paccamonti D, Godke RA. Preservation of equine sperm stored in the epididymides at 4°C for 24, 48, 72 and 96 Hours. Theriogenology. 2002;58:401–4.

    Article  Google Scholar 

  10. Yu I, Leibo SP. Recovery of motile, membrane-intact spermatozoa from canine epididymides stored for 8 days at 4°C. Theriogenology. 2002;57:1179–90.

    Article  CAS  PubMed  Google Scholar 

  11. Fournier-Delpech S, Colas G, Courot M, Ortavant R, Brice G. Epididymal sperm maturation in the ram: motility, fertilizing ability and embryonic survival after uterine artificial insemination. Ann. Biol. Anim. Bioch. Biophys. 1979;19(3A):597–605.

    Article  Google Scholar 

  12. Niwa K, Ohara K, Hosoi Y, Iritani A. Early events of in-vitro fertilization of cat eggs by epididymal spermatozoa. J. Reprod. Fertil. 1985;74(2):657–60.

    CAS  PubMed  Google Scholar 

  13. Temple-Smith PD, Southwick GJ, Yates CA, Trounson AO, de Kretser DM. Human pregnancy by in vitro fertilization (IVF) using sperm aspirated from the epididymis. J. In Vitro Fertil. Embryo Transfer. 1985;2:119–222.

    Article  CAS  Google Scholar 

  14. Marks SL, Dupuis J, Mickelsen WD, Memon MA, Platz CC. Conception by use of postmortem epididymal semen extraction in a dog. J. Anim. Vet. Med. Assoc. 1994;204(10):1639–40.

    CAS  Google Scholar 

  15. Graff KJ, Chandler JE, Reggio BC, Lim JM, Canal A, Carter JA, Meintjes M, Godke RA: Pregnancies obtained from IVF with noncapacitated epididymal bovine spermatozoa. Proc. 3rd Intern’l. Mtg. Biotech. Anim. Reprod. Cairo, Egypt (Nov. 2–6). 1996: pp. 19–21.

  16. Bonduel M, Wilikens A, Buysse A, Van Assche E, Devroey P, Van Steirteghem AC, et al. A follow-up study of children born after intracytoplasmic sperm injection (ICSI) with epididymal and testicular spermatozoa and after replacement of cryopreserved embryos obtained after ICSI. Hum. Reprod. 1998;13(Suppl 1):196–207.

    Google Scholar 

  17. Morrell JM, Nubbemeyer R, Heistermann M, Rosenbusch J, Kuderling I, Holt W, et al. Artificial insemination in Callithrix jacchus using fresh or cryopreserved sperm. Anim. Reprod. Sci. 1998;52(2):165–74.

    Article  CAS  PubMed  Google Scholar 

  18. Bartels P, Lubbe K, Kilian I, Friedmann Y, van Dyk G, Mortimer D. In vitro maturation and fertilization of lion (Panthera leo) oocytes using frozen-thawed epididymal spermatozoa recovered by cauda epididymectomy of an immobilized lion. Theriogenology. 2000;53(1):325. abstract.

    Google Scholar 

  19. Kusunoki H, Daimaru H, Miami S, Nishimoto S, Yamane K-I, Fukumoto Y. Birth of a chimpanzee (Pan troglodytes) after artificial insemination with cryopreserved epididymal spermatozoa collected postmortem. Zoo. Biol. 2001;20:135–43.

    Article  Google Scholar 

  20. Ikeda H, Kikuchi K, Noguchi J, Takeda H, Shimada A, Mizokami T, et al. Effect of preincubation of cryopreserved porcine epididymal sperm. Theriogenology. 2002;57:1309–18.

    Article  CAS  PubMed  Google Scholar 

  21. Martins CF, Rumpf R, Pereira DC, Dode MN. Cryopreservation of epididymal bovine spermatozoa from dead animals and its use in in-vitro embryo production. Anim. Reprod. Sci. 2007;101:326–31.

    Article  CAS  PubMed  Google Scholar 

  22. Said S, dan Saili T. Rescuing genetic material of unexpectedly die animal. Jurnal Ilmu Ternak dan Veteriner. 2007;12(2):147–52.

    Google Scholar 

  23. World health Organization: WHO Laboratory manual for the examinations of human semen and sperm-cervical mucus interaction (3e). Cambridge; The press syndicated of the University of Cambridge. 1997.

  24. Sidhu KS, Guraya SS: In: Buffalo bull semen morphology, biochemistry, physiology and methodology. USA publishers and distributors, Ludhiana, India. 1985a; 152–154.

  25. Jeyendran RS, Vandervent HH, Perez-Paleac ZM, Crabo BG, Zaneveld LJD. Development of an assay to assess the functional integrity of human sperm membrane and its relationship to other semen characteristics. J. Reprod. Fertil. 1984;70:219–25.

    Article  CAS  PubMed  Google Scholar 

  26. Duncan DB. Multiple range and multiple F tests. Biometrics. 1955;11:1–42.

    Article  Google Scholar 

  27. Scott TW, Voglmayer JK, Setchell BP. Lipid composition and metabolism in testicular and ejaculated spermatozoa. The Biochemical Journal. 1967;102:456–60.

    CAS  PubMed  Google Scholar 

  28. Arora R, Dinakar N, Prasad MRN. Biochemical changes in the spermatozoa and luminal contents of different regions of the epididymis of the rhesus monkey, Macaca mulalta. Contraception. 1975;11:689–700.

    Article  CAS  PubMed  Google Scholar 

  29. Hammerstedt RH, Keith AD, Hay S, Deluca N, Amann RP. Changes in ram sperm membrane during epididymis transit. Arch. Biochem. Biophys. 1979;196:7–12.

    Article  CAS  PubMed  Google Scholar 

  30. Thun R, Hurtado M, Janett F. Comparison of Biociphos-plus and TRIS-egg yolk extender for cryopreservation of bull semen. Theriogenology. 2002;57:1087–94.

    Article  PubMed  Google Scholar 

  31. Mancini A, Marinis LD, Oradei A, Hallgass ME, Conte G, Pozza D, et al. Coenzyme Q10 concentration in normal and pathological Human seminal fluid’s. J. Androl. 1994;15(6):591–4.

    CAS  PubMed  Google Scholar 

  32. Jones R, Mann T. Damage to ram spermatozoa by peroxidation of endogenous phospholipids. J.Reprod. Fertil. 1977;50:261–8.

    CAS  PubMed  Google Scholar 

  33. Ernster L, Forsneark-Andree P. Ubiquinol: an endogenous antioxidant in aerobic organisms. Clin. Invest. 1993;71:S60–65.

    Article  CAS  Google Scholar 

  34. Ernster L, Dallner G. Biochemical, physiological and medical aspects of ubiquinone function. Biochem. Biophys. Acta. 1995;1271:195–204.

    PubMed  Google Scholar 

  35. Lewin A, Lavon H. The effect of Coenzyme Q10 on sperm motility and function. Molecular Aspects of Medicine. 1997;18(Suppl):S 213–9.

    CAS  Google Scholar 

  36. Thomas SR, Neuzil J, Stocker R. Co supplementation with coenzyme Q prevents the prooxidant effects of alphatocopherol and increase the resistance of LDL to transition metal-dependent oxidation initiation. Arterioscler. Thromb. Vasc. Biol. 1996;16:687–96.

    CAS  PubMed  Google Scholar 

  37. Thomas SR, Neuzil J, Stocker R. Inhibition of LDL oxidation by ubiquinone-10. A protective mechanism for coenzyme Q in atherogenesis? Mol.Aspects. Med. 1997;18:S85–103.

    Article  CAS  PubMed  Google Scholar 

  38. Booth NH, McDonald LE. Veterinary pharmacology and Therapeutics. 6th ed. New Delhi: Kalyani publisher; 1982.

    Google Scholar 

  39. Mazzilli F, Cerasaro M, Bisanti A, Rossi T, Dondero F: Seminal parameters and the swelling test in patients with sperm before and after treatment with ubiquinone (CoQ10). 2nd international symposium on reproductive medicine. Acta Medica, Edizioni e Congresi, Rome, Italy: Fiuggi. 1988; 71.

  40. Ball BA, Vo AT, Baumber J. Reactive oxygen species generation by equine spermatozoa. Am. J. Vet. Res. 2001;62:5508–15.

    Article  Google Scholar 

  41. Hammerstedt RH, Graham JK, Nolan JP. Cryopreservation of mammalian sperm: What we ask them to survive. J. Androl. 1990;11:73–88.

    CAS  PubMed  Google Scholar 

  42. Riddle VM, Lorenz N. Nonenzymic formulation of toxic levels of methylglyoxal from glycerol and dihydroxyacetone in Ringers phosphate suspensions of avian spermatozoa. Biochem, Biophy. Res. Commun. 1973;50:27–34.

    Article  CAS  Google Scholar 

  43. Fahy GM, Lilley TH, Linsdell H, Douglas MS, Meryman HT. Cryoprotectant toxicity and cryoprotectant toxicity reduction: In search of molecular mechanisms. Cryobiol. 1990;27:247–68.

    Article  CAS  Google Scholar 

  44. Watson PF. The effects of cold shock on sperm cell membranes. In: Morris GJ, Clark A, editors. Effects of Low temperatures on Biological membranes. London: Academic; 1981. p. 189–218.

    Google Scholar 

  45. Rodojcic L, Vukotic-Maletic V, Balint B. Current Knowledge on cryopreservation of spermattzoa, ovum cells and zygotes. Medicinski pregled. 1998;51(1–2):29–36.

    Google Scholar 

  46. Curry MR, Watson PF. Osmotic effects on ram and human sperm membranes in relation to thawing injury. Cryobiology. 1994;31(1):39–46.

    Article  CAS  PubMed  Google Scholar 

  47. Watson PF. Recent developments and concepts in the cryopreservation of spermatozoa and the assessment of their post-thawing function. Reprod. Fertility. Dev. 1995;7:747–869.

    Article  Google Scholar 

  48. Harrison RAP, White IG. Glycolytic enzymes in the spermatozoa and cytoplasmic droplets of bull, boar and ram and their leakage after shock. J Reprod Fertil. 1972;30:105–15.

    Article  CAS  PubMed  Google Scholar 

  49. Simpson AM, White IG. Effect of cold shock and cooling rate on calcium uptake of ram spermatozoa. Anim. Reprod. Sci. 1986;12:131–43.

    Article  CAS  Google Scholar 

  50. Robertson L, Bailey JL, Buhr MM. Effects of cold shock and phospholipase A2 on intact boar spermatozoa and sperm head plasma membranes. Mol. Reprod Dev. 1990;26:143–9.

    Article  CAS  PubMed  Google Scholar 

  51. Cotran RS, Kumar V, Robbins SL. Robbin’s Pathologic basis of disease. 4th ed. Philadelphia: WB Saunders Co; 1989. p. 9–16.

    Google Scholar 

  52. Bilodeau JF, Chatterjee S, Sired MA, Gagnon C. Levels of antioxidant defenses are decreased in bovine spermatozoa after a cycle of freezing and thawing. Mol. Reprod. Dev. 2000;55:282–8.

    Article  CAS  PubMed  Google Scholar 

  53. Quinn PJ. Principles of membrane stability and phase behavior under extreme conditions. J Bioenerg Biomembr. 1989;21:3–19.

    Article  CAS  PubMed  Google Scholar 

  54. Holt WV, North RD. The role of membrane —active lipids in the protection of ram spermatozoa during cooling and storage. Gamete Res. 1988;19:77–89.

    Article  CAS  PubMed  Google Scholar 

  55. De Leeuw FE, Chen HC, Colenbrander B, Verkleji AJ. Cold-induced ultra structural changes in bull and boar sperm plasma membranes. Cryobiology. 1990;27:171–83.

    Article  PubMed  Google Scholar 

  56. Buhr MM, Curtis EF, Kakuda NS. Composition and behavior of head membrane lipids of fresh and cryopreserved boar sperm. Cryobiology. 1994;31:224–38.

    Article  CAS  PubMed  Google Scholar 

  57. White IG. Lipids and calcium uptake of sperm in relation to cold shock and preservation: A review. Reproduction. Fertility. Dev. 1993;5(6):639–58.

    Article  CAS  Google Scholar 

  58. Aitken RJ. Free radicals, lipid peroxidation and sperm function. Reprod. Fertil. Dev. 1995;7:659–68.

    Article  CAS  PubMed  Google Scholar 

  59. Drobinis EZ, Crowe LM, Berger T, Anchordoguy TJ, Overstreet JW, Crowe JH. Cold shock damage is due to lipid phase transitions in cell-membranes-a demonstration using sperm as a model. J. Exp. Zool. 1993;265:432–7.

    Article  Google Scholar 

  60. Rao B, Soufir JC, Martin M. Lipid peroxidation in human spermatozoa as related to mid piece abnormalities and motility. Gamete Res. 1989;24:127–34.

    Article  CAS  PubMed  Google Scholar 

  61. Aitken RJ, West K, Buckingham DW. Leukocytic infiltration into the human ejaculate and its association with semen quality, oxidative stress, and sperm function. J. Androl. 1994;15:343–52.

    CAS  PubMed  Google Scholar 

  62. Baumber J, Ball BA, Gravance CG, Medina V, Davies-Morel MC. The effect of reactive species on equine sperm motility, viability, acrosomal integrity, mitochondria membrane potential, and membrane lipid peroxidation. J.Androl. 2001;21:895–902.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Veterinary Gynaecology & Obstetrics, Faculty of Veterinary and Animal Sciences West Bengal University of ANIMAL and Fishery Sciences, 37and 68, Kshudiram Bose Sarani, Belgachia, Kolkata, 700037, West Bengal, India

    Uttam Datta, M. Chandra Sekar & Manik Lal Hembram

  2. Department of Animal Genetics and Breeding, Faculty of Veterinary and Animal Sciences West Bengal University of ANIMAL and Fishery Sciences, 37and 68, Kshudiram Bose Sarani, Belgachia, Kolkata, 700037, West Bengal, India

    Raju Dasgupta

Authors
  1. Uttam Datta
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. Chandra Sekar
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Manik Lal Hembram
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Raju Dasgupta
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Uttam Datta.

Additional information

Capsule In this method spermatozoa remaining within ligated cauda epididymides were treated with electrolyte free media and freezed at -10°C and were assessed after thawing.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Datta, U., Chandra Sekar, M., Hembram, M.L. et al. Development of a new method to preserve caprine cauda epididymal spermatozoa in-situ at -10°C with electrolyte free medium. J Assist Reprod Genet 26, 467–473 (2009). https://doi.org/10.1007/s10815-009-9344-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10815-009-9344-4

Keywords

Search

Navigation