Vet Med - Czech, 2004, 49(1):1-8 | DOI: 10.17221/5668-VETMED

Bull and boar sperm DNA integrity evaluated by sperm chromatin structure assay in the Czech Republic

R. Rybak, L. Faldikova, M. Faldyna, M. Machatkova, J. Rubes
Veterinary Research Institute, Brno, Czech Republic

Analysis of sperm parameters is very important for predicting the outcome of assisted reproductive techniques and is necessary for determination of fertility potential of males tested for artificial insemination. In our study we have determined the level of bull and boar sperm DNA damage by Sperm Chromatin Structure Assay (SCSA). This test is based on increased susceptibility of altered DNA (strand breaks) in sperm nuclear chromatinto in situ denaturation measured by flow cytometry after staining with acridine orange (AO). Sperm chromatin damage was quantified by percentages of spermatozoa with detectable DNA Fragmentation Index - DFI divided into moderate (m-DFI) and high (h-DFI) DFI. Percentage of immature cells (HDS; cells with High DNA Stainability) was also evaluated. We measured sperm SCSA parameters in a total of 37 bulls in two groups from different localities and 68 boar samples from one locality. Significantly higher percentage of spermatozoa with detectable DFI was detected in six bulls (16.2%) and a significantly higher percentage of immature cell forms (HDS) was found in other six bulls (16.2%) among all tested bulls. The mean percentages of spermatozoa with h-DFI and HDS of bulls from the second group were statistically higher than those from the first group (P < 0.01). Five boars (7.4%) of all tested boars had significantly higher percentage of spermatozoa with DFI and 18 boars (26.5%) had significantly higher percentage of sperm with HDS compared to the other boars. Both percentages of spermatozoa with DFI and HDS were significantly higher in one boar compared to the others. Boars had significantly higher percentages of spermatozoa with h-DFI and HDS (P < 0.0001) in comparison to bulls. For individual bulls, the highest percentages of spermatozoa with DFI and HDS were 20.8% and 3.5%, respectively while for boars these were 17.6% and 10.2%, respectively. No significant correlations were found between percentages of spermatozoa with DFI and HDS. This sensitive procedure seems to be convenient as additional method for semen quality detection in farm animals before their exploitation in breeding.

Keywords: sperm; DNA integrity; flow cytometry; bull; boar; fertility

Published: January 31, 2004  Show citation

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Rybak R, Faldikova L, Faldyna M, Machatkova M, Rubes J. Bull and boar sperm DNA integrity evaluated by sperm chromatin structure assay in the Czech Republic. Vet Med - Czech. 2004;49(1):1-8. doi: 10.17221/5668-VETMED.
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References

  1. Ahmadi A., Soon-Chye N.G. (1999): Fertilizing ability of DNA damaged spermatozoa. J. Exp. Zool., 284, 696-704. Go to original source...
  2. Ballachey B.E., Hohenboken W.D., Evenson D.P. (1987): Heterogeneity of sperm nuclear chromatin structure and its relationship to fertility of bulls. Biol. Reprod., 36, 915-925. Go to original source... Go to PubMed...
  3. Bochenek M., Smorag Z., Pilch J. (2001): Sperm chromatin structure assay of bulls qualified for artificial insemination. Theriogenology, 56, 557-567. Go to original source... Go to PubMed...
  4. Chacon J., (2001): Assessment of sperm morphology in zebu bulls, under field conditions in the tropics. Reprod. Domest. Anim., 36, 91-99. Go to original source... Go to PubMed...
  5. Colas G. (1983): Factors affecting the quality of ram semen. In: Haresign W. (ed.): Sheep Production. Bu erworths, Stoneham, Massachuse s. 285-291.
  6. Engh E., Clausen O.P., Scholberg A., Tollefsrud A., Purvis K. (1992): Relationship between sperm quality and chromatin condensation measured by sperm DNA fluorescence using flow cytometry. Int. J. Androl., 15, 407-415. Go to original source... Go to PubMed...
  7. Evenson D.P., Jost L.K. (1994): Sperm chromatin structure assay: DNA denaturability. In: Darzynkiewicz Z., Robinson J.P., Crissman H.A. (eds.): Methods in Cell Biology. Vol 42, Flow Cytometry, Second Edition. Academic Press, Inc., Orlando. 159-176. Go to original source...
  8. Evenson D.P., Jost L.K. (2000): Sperm chromatin structure assay is useful for fertility assessment. Method Cell Sci., 22, 169-189. Go to original source... Go to PubMed...
  9. Evenson D.P., Melamed M.R. (1983): Rapid analysis of normal and abnormal cell types in human semen and testis biopsies by flow cytometry. J. Histochem. Cytochem., 31, 248-253. Go to original source... Go to PubMed...
  10. Evenson D.P., Darzynkiewicz Z., Melamed M.R. (1980): Relation of mammalian sperm chromatin heterogenity to fertility. Science, 210, 1131-1133. Go to original source... Go to PubMed...
  11. Evenson D.P., Lee J., Darzynkiewicz Z., Melamed M.R. (1985): Rhodamine 123 alters the mitochondrial ultrastructure of cultured L 1210 cells. J. Histochem. Cytochem., 33, 353-359. Go to original source... Go to PubMed...
  12. Evenson D.P., Jost L.K., Gandy J. (1993a): Effect of methyl methansulfonate on mouse sperm chromatin structure and testicular cell kinetics. Environ. Mol. Mutagen., 21, 144-153. Go to original source... Go to PubMed...
  13. Evenson D.P., Parks J.E., Kaproth M.T., Jost L.K. (1993b): Rapid determination of sperm cell concentration in bovine semen by flow cytometry. J. Dairy. Sci., 76, 86-84. Go to original source... Go to PubMed...
  14. Evenson D.P., Jost L.K., Marshall D., Zinaman M.J., Clegg E., Purvis K., de Angelis P., Claussen O.P. (1999): Utility of sperm chromatin structure assay as a diagnostic tool in the human fertility clinic. Hum. Reprod., 14, 1039-1049. Go to original source... Go to PubMed...
  15. Evenson D.P., Larson K.L., Jost L.K. (2002): Sperm chromatin structure assay: Its clinical use for detecting sperm DNA fragmentation in male infertility and comparisons with other techniques. J. Androl., 23, 25-43. Go to original source... Go to PubMed...
  16. Evenson D.P., Larson-Cook K.L., Virro M. (2003): Sperm chromatin structure assay (SCSA) parameters relationship with fertilization rate, blastocyst development and pregnancy outcomes following in vitro fertilization (IVF) and IVF with intracytoplasmic sperm injection (ICSI). Fertil. Steril., 80 (S3), 113-114. Go to original source...
  17. Garner D.L., Pinkel D., Johnson L.A., Pace M.M. (1986): Assessment of spermatozoal function using dual fluorescein staining and flow cytometric analyses. Biol. Reprod., 34, 127-138. Go to original source... Go to PubMed...
  18. Giwercman A., Richthoff J., Hjollund H., Bonde J.P., Jepson K., Frohm B., Spano M. (2003): Correlation between sperm motility and sperm chromatin structure assay parameters. Fertil. Steril., 80, 1404-1412. Go to original source... Go to PubMed...
  19. Gogol P., Bochenek M., Smorag Z. (2002): Effect of rabbit age on sperm chromatin structure. Reprod. Domest. Anim., 37, 92-95. Go to original source... Go to PubMed...
  20. Graham J.K., Kunze E., Hammerstedt R.H. (1990): Analyses of sperm viability, acrosomal integrity and mitochondrial function using flow cytometry. Biol. Reprod., 43, 55-64. Go to original source... Go to PubMed...
  21. Johnson L.A., Flook J., Hawk H., Look M., Pinkel D. (1987): Flow sorting of X and Y chromosome bearing spermatozoa into two populations. Gamete Res., 16, 1-9. Go to original source... Go to PubMed...
  22. Johnson L.A., Flook J., Hawk H. (1989): Sex preselection in rabbits: Live births from X and Y sperm separated by DNA and cell sorting. Biol. Reprod., 41, 199-203. Go to original source... Go to PubMed...
  23. Johnson K.R., Dewey C.E., Bobo J.K., Kelling C.L., Lunstra D.D. (1998): Prevalence of morphologic defects in spermatozoa from beef bulls. J. Am. Vet. Med. Assoc., 213, 1468-1471. Go to original source... Go to PubMed...
  24. Kruger T.F., Menkveld R., Stander F.S. (1986): Sperm morphologic features as a prognostic factor in in vitro fertilization. Fertil. Steril., 46, 1118-1123. Go to original source... Go to PubMed...
  25. Larson-Cook K.L, Brannian J.D., Hansen K.A., Kasperson K.M., Aamold E.T., Evenson D.P. (2003): Relationship between the outcomes of assisted reproductive techniques and sperm DNA fragmentation as measured by the sperm chromatin structure assay. Fertil. Steril., 80, 895-902. Go to original source... Go to PubMed...
  26. Larson K.L., DeJonge C., Barnes A.M., Jost L.K., Evenson D.P. (2000): Sperm chromatin structure assay parameters as predictors of failed pregnancy following assisted reproductive techniques. Hum. Reprod., 15, 1717-1722. Go to original source... Go to PubMed...
  27. Lopes S., Jurisicova A., Casper R.F. (1998): Gamete specific DNA fragmentation in unfertilized human oocytes a er intracytoplasmic sperm injection. Hum. Reprod., 13, 703-708. Go to original source... Go to PubMed...
  28. Love C.C., Thompson J.A., Lowry V.K., Varner D.D. (2002): Effect of storage time and temperature on stallion sperm DNA and fertility. Theriogenology, 57, 1135-1142. Go to original source... Go to PubMed...
  29. Ochsendorf F.R. (1999): Infections in the male genital tract and reactive oxygen species. Hum. Reprod., 5, 399-420. Go to original source... Go to PubMed...
  30. Pacifici R., Altieri I., Gandini L., Lenzi A., Pachini S., Rosa M., Zuccaro P., Dondero F. (1993): Nicotine, cotinine and trans-3-hydroxycotinine levels in seminal plasma of smokers: effects on sperm parameters. Ther. Drug. Monit., 15, 358-363. Go to original source... Go to PubMed...
  31. Perreault S.D., Rubes J., Robbins W.A., Evenson D.P., Selevan S.G. (2000): Evaluation of aneuploidy and DNA damage in human spermatozoa: applications in field studies. Andrologia, 32, 247-254. Go to original source... Go to PubMed...
  32. Po s R.J., Newbury C.J., Smith G., Notarianni L.J., Jefferies T.M. (1999): Sperm chromatin damage associated with male smoking. Mutat. Res., 423, 103-111. Go to original source... Go to PubMed...
  33. Sailer B.L., Jost L.K., Evenson D.P. (1995a): Mammalian sperm DNA susceptibility to in situ denaturation associated with the presence of DNA strand breaks as measured by the terminal deoxynukleotidyl transferase assay. J. Androl., 16, 80-87. Go to original source... Go to PubMed...
  34. Sailer B.L., Jost L.K., Erickson K.R., Tajiran M.A., Evenson D.P. (1995b): Effects of X-irradiation on mouse testicular cells and sperm chromatin structure. Environ. Mol. Mutagen., 25, 23-30. Go to original source... Go to PubMed...
  35. Sailer B.L., Sarkar L.J., Bjordahl J.A., Jost L.K., Evenson D.P. (1997): Effects of heat stress on mouse testicular cells and sperm chromatin structure. J. Androl., 18, 294-301. Go to original source... Go to PubMed...
  36. Selevan S.G., Borkovec L., Slo V.L., Zudová Z., Rubeš J., Evenson D.P., Perreault S.D. (2000): Semen quality and reproductive health of young Czech men exposed to seasonal air pollution. Environ. Health. Perspect., 108, 887-894. Go to original source... Go to PubMed...
  37. Setchell B.P., D'Occhio M.J., Hall M.J., Laurie M.S., Tucker M.J., Zupp J.L. (1988): Is embryonic mortality increased in normal female rats mated to subfertile males? J. Reprod. Fertil., 82, 567-574. Go to original source... Go to PubMed...
  38. Sills E.S., Fryman J., Perloe M., Michels K., Tucker M. (2004): Chromatin fluorescence characteristic and standard semen analysis parameters: correlations observed in andrology testing among 136 males referred for infertility evaluation. Obstet. Gynaecol., 24, 74-77. Go to original source... Go to PubMed...
  39. Spano M., Cordelli E., Leter G., Lombardo F., Lenzi A., Gandini L. (1999): Nuclear chromatin variations in human spermatozoa undergoing swim-up and cryopreservation evaluated by the flow cytometric sperm chromatin structure assay. Mol. Hum. Reprod., 5, 29-37. Go to original source... Go to PubMed...
  40. Spano M., Bonde J.P., Hjollund H.I., Kolstad H.A., Cordelli E., Leter G. (2000): Sperm chromatin damage im

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