Abstract
Semen analysis is a basic step in the investigation of several disturbances affecting the male genital tract. Analysis of seminal parameters provides important clinical information on the spermatogenesis and functional competence of spermatozoa, as well as on the secretory pattern of the accessory genital glands. Semen analysis is particularly useful in the evaluation of couples requiring fertility investigation (to detect genital infections and pathologies) and in verifying the influence of environmental factors, drugs, lifestyle, chemical products, and professional activities on several diseases affecting male reproductive health. Measure of semen quality is of substantial interest for diagnoses in clinical urology, andrology, and gynecology. Currently, basic requirements for semen analysis are standardized by World Health Organization (WHO) guidelines that describe several procedures for an objective evaluation of the semen quality with diagnostic purposes. These guidelines include: parameters for the physical and biochemical evaluation of semen; parameters for the analysis of sperm characteristics; and other seminal parameters that can be easily adopted in any laboratory. This report summarizes current concepts on semen analysis and the significance of the seminal parameters for reaching a diagnosis based on the procedures recommended by WHO guidelines. J. Clin. Lab. Anal. 17:247–258, 2003. © 2003 Wiley‐Liss, Inc.
Keywords: male fertility, spermatozoa, semen analysis, spermatogenesis, accessory sex glands, genital infections, antisperm antibodies
REFERENCES
- 1. Comhaire F, Vermeulen L. Human semen analysis. Hum Reprod Update 1995;1:343–362. [DOI] [PubMed] [Google Scholar]
- 2. Silber SJ. Evaluation and treatment of male infertility. Clin Obstet Gynecol 2000;43:854–888. [DOI] [PubMed] [Google Scholar]
- 3. Sharpe RM. Environment, lifestyle and male infertility. Baillieres Best Pract Res Clin Endocrinol Metab 2000;14:489–503. [DOI] [PubMed] [Google Scholar]
- 4. Bonde JP, Storgaard L. How work‐place conditions, environment toxicants and lifestyle affect male reproductive function. Int J Androl 2002;25:262–268. [DOI] [PubMed] [Google Scholar]
- 5. Barratt CL, St John JC. Diagnostic tools in male infertility. Hum Reprod 1998;13(suppl 1):51–61. [DOI] [PubMed] [Google Scholar]
- 6. Carrell DT. Semen analysis at the turn of the century: an evaluation of potential uses of new sperm function assays. Arch Androl 2000;44:65–75. [DOI] [PubMed] [Google Scholar]
- 7. World Health Organization . WHO laboratory manual for the examination of human semen and sperm‐cervical mucus interaction. Cambridge: Cambridge University Press; 1999. p 1–128 [Google Scholar]
- 8. Johnson L, Staub C, Neaves WB, Yanagimachi R. Live human germ cells in the context of their spermatogenic stages. Hum Reprod 2001;16:1575–1582. [DOI] [PubMed] [Google Scholar]
- 9. Clermont Y. The cycle of the seminiferous epithelium in man. Am J Anat 1963;112:35–51. [DOI] [PubMed] [Google Scholar]
- 10. de Kretser DM, Loveland KL, Meinhardt A, Simorangkir D, Wreford N. Spermatogenesis. Hum Reprod 1998;13():1–suppl 1 8. [DOI] [PubMed] [Google Scholar]
- 11. Hecht NB. Molecular mechanisms of male germ cell differentiation. Bioessays 1998;20:555–561. [DOI] [PubMed] [Google Scholar]
- 12. Griswold MD. The central role of Sertoli cells in spermatogenesis. Semin Cell Dev Biol 1998;9:411–416. [DOI] [PubMed] [Google Scholar]
- 13. McLachlan RI. The endocrine control of spermatogenesis. Baillieres Best Pract Res Clin Endocrinol Metab 2000;14:345–362. [DOI] [PubMed] [Google Scholar]
- 14. Grootegoed JA, Siep M, Baarends WM. Molecular and cellular mechanisms in spermatogenesis. Baillieres Best Pract Res Clin Endocrinol Metabol 2000;14:331–343. [DOI] [PubMed] [Google Scholar]
- 15. Amory JK, Bremner W. Endocrine regulation of testicular function in men: implications for contraceptive development. Mol Cell Endocrinol 2001;182:175–179. [DOI] [PubMed] [Google Scholar]
- 16. Andrade‐Rocha FT. Study on the general characteristics of the spermatozoa in polyzoospermic semen. Rev Bras Ginec Obstet 1994;16:77–79. [Google Scholar]
- 17. Chan SYW, Tang LCH, Tang GWK, Ho PC, Wang C. Spermatozoal fertilizing capacity in polyzoospermia: a preliminary study. Andrologia 1986;18:208–213. [DOI] [PubMed] [Google Scholar]
- 18. Calamera JC, Giovenco P, Brugo S, Dondero F, Nicholson RF. Adenosine 5‐triphosphate (ATP) content and acrosine activity in polyzoospermic subjects. Andrologia 1987;19:460–463. [DOI] [PubMed] [Google Scholar]
- 19. Töpfer‐Petersen E, Völcker C, Heissler E, Schill WB. Absence of acrosome reaction in polyzoospermia. Andrologia 1987;19:225–228. [DOI] [PubMed] [Google Scholar]
- 20. Glezerman M, Bernstein D, Zakut C, Misgav N, Insler V. Polyzoospermia: a definite pathologic entity. Fertil Steril 1982;38:605–608. [DOI] [PubMed] [Google Scholar]
- 21. Tournaye H, Staessen C, Camus M, Verheyen G, Devroey P, van Steirteghem A. No evidence for a decreased fertilizing potential after in‐vitro fertilization using spermatozoa from polyzoospermic men. Hum Reprod 1997;10:2183–2185. [DOI] [PubMed] [Google Scholar]
- 22. Merino Ruiz MC, De Leon Cervantes MG, Garcia Flores RF. Male sterility and its association with genital disease and environmental factors. Ginecol Obstet Mex 1995;63:427–431. [PubMed] [Google Scholar]
- 23. Forti G, Krausz C. Clinical review 100: evaluation and treatment of the infertile couple. J Clin Endocrinol Metab 1998;83:4177–4188. [DOI] [PubMed] [Google Scholar]
- 24. Dohle GR, Halley DJ, Van Hemel JO, et al. Genetic risk factors in infertile men with severe oligozoospermia and azoospermia. Hum Reprod 2002;17:13–16. [DOI] [PubMed] [Google Scholar]
- 25. Matorras R, Diez J, Corcostegui B, et al. Spontaneous pregnancy in couples waiting for artificial insemination donor because of severe male infertility. Eur J Obstet Gynecol Reprod Biol 1996;70:175–178. [DOI] [PubMed] [Google Scholar]
- 26. Kolettis PN. The evaluation and management of the azoospermic patient. J Androl 2002;23:293–305. [PubMed] [Google Scholar]
- 27. Daudin M, Bieth E, Bujan L, Massat G, Pontonnier F, Mieusset R. Congenital bilateral absence of the vas deferens: clinical characteristics, biological parameters, cystic fibrosis transmembrane conductance regulator gene mutations, and implications for genetic counseling. Fertil Steril 2000;74:1164–1174. [DOI] [PubMed] [Google Scholar]
- 28. Moench GL, Holt H. Sperm morphology in relation to fertility. Am J Obstet Gynecol 1931;22:199–210. [Google Scholar]
- 29. Hammen R. Studies on impaired fertility in man with special reference to the male. Acta Obstet Gynecol Scand 1944;24 (suppl 3):1–205. [Google Scholar]
- 30. MacLeod J. Human seminal cytology as a sensitive indicator of the germinal epithelium. Int J Fertil 1964;9:281–295. [PubMed] [Google Scholar]
- 31. Eliasson R. Standards for investigation of human semen. Andrologia 1971;3:49–64. [Google Scholar]
- 32. David G, Bisson JP, Czyglik F, Jouannet P, Gernigon C. Anomalies morphologiques du spermatozöide humain. 1) Proposition pour un système de classification. J Gynecol Obstet Biol Reprod (Paris) 1975;4(supp 1):17–36. [Google Scholar]
- 33. Hartman CG, Schoenfeld C, Copeland E. Individualism in semen picture of infertile men. Fertil Steril 1964;15:231–253. [DOI] [PubMed] [Google Scholar]
- 34. Zamboni L. The ultrastructural pathology of the spermatozoon as a cause of infertility: the role of electron microscopy in the evaluation of semen quality. Fertil Steril 1987;48:711–734. [DOI] [PubMed] [Google Scholar]
- 35. Bartoov B, Eltes F, Weissenberg R, Lunenfeld B. Morphological characterization of abnormal human spermatozoa using transmission electron microscopy. Arch Androl 1980;5:305–322. [DOI] [PubMed] [Google Scholar]
- 36. Kruger TF, Menkveld R, Stander FSH, et al. Sperm morphologic features as a prognostic factor in in vitro fertilization. Fertil Steril 1986;46:1118–1123. [DOI] [PubMed] [Google Scholar]
- 37. Mundy AJ, Ryder TA, Edmonds DK. Morphometric characteristics of motile spermatozoa in subfertile men with an excess of non‐sperm cells in the ejaculate. Hum Reprod 1994;9:1701–1704. [DOI] [PubMed] [Google Scholar]
- 38. Salanova M, Gandini L, Lenzi A, et al. Is hyperdiploidy of immature ejaculated germ cells predictive of testis malignancy? A comparative study in healthy normozoospermic, infertile and testis tumor suffering subjects. Lab Invest 1999;79:1127–1135. [PubMed] [Google Scholar]
- 39. Arata de Bellabarba G, Tortolero I, Villarroel V, Molina CZ, Bellabarba C, Velasquez E. Nonsperm cells in human semen and their relationship with semen parameters. Arch Androl 2000;45:131–136. [DOI] [PubMed] [Google Scholar]
- 40. Ariagno J, Cury S, Mendeluk G, et al. Shedding of immature germ cells. Arch Androl 2002;48:127–131. [DOI] [PubMed] [Google Scholar]
- 41. Couture M, Ulstein M, Leonard J, Paulsen CA. Improved staining method for differentiating of immature germ cells from white blood cells in human seminal fluid. Andrologia 1976;8:61–66. [DOI] [PubMed] [Google Scholar]
- 42. Homyk M, Anderson DJ, Wolff H, Herr JC. Differential diagnosis of immature germ cells in semen utilizing monoclonal antibody MHS‐10 to the intra‐acrosomal antigen SP‐10. Fertil Steril 1990;53:323–330. [DOI] [PubMed] [Google Scholar]
- 43. Amer M, Abd Elnasser T, El Haggar S, Mostafa T, Abdel‐Malak G, Zohdy W. May‐Grunwald‐Giemsa stain for detection of spermatogenic cells in the ejaculate: a simple predictive parameter for successful testicular sperm retrieval. Hum Reprod 2001;16:1427–1432. [DOI] [PubMed] [Google Scholar]
- 44. Cooper TG. Epididymis and sperm function. Andrologia 1996;28:57–59. [PubMed] [Google Scholar]
- 45. Moore HDM. Contribution of epididymal factors to sperm maturation and storage. Andrologia 1998;30:233–239. [DOI] [PubMed] [Google Scholar]
- 46. Potts RJ, Jefferies TM, Notarianni LJ. Antioxidant capacity of the epididymis. Hum Reprod 1999;14:2513–2516. [DOI] [PubMed] [Google Scholar]
- 47. Krause W, Bohring C. Why do we determine alpha‐glucosidase activity in human semen during infertility work‐up? Andrologia 1999;31:289–294. [DOI] [PubMed] [Google Scholar]
- 48. Comhaire F, Mahmoud A, Schoonjans F, Kint J. Why do we continue to determine α‐glucosidase in human semen? Andrologia 2002;34:8–10. [DOI] [PubMed] [Google Scholar]
- 49. Lundquist F. Aspects of the biochemistry of human semen. Acta Obstet Gynecol Scand 1949;19(suppl(:1–105. [Google Scholar]
- 50. Bukovsky A, Thaler CJ, McIntyre JA. Antigens of immunoglobulin G‐Fc receptor III in human male reproductive tract accessory glands. 1991;Fertil Steril 55:595–602. [PubMed] [Google Scholar]
- 51. Robert M, Gagnon C. Sperm motility inhibitor from human seminal plasma: association with semen coagulum. Hum Reprod 1995;10:2192–2197. [DOI] [PubMed] [Google Scholar]
- 52. Gonzales GF. Function of seminal vesicles and their role on male infertility. Asian J Androl 2001;3:251–258. [PubMed] [Google Scholar]
- 53. de Lamirande E, Yoshida K, Yoshiike TM, Iwamoto T, Gagnon C. Semenogelin, the main protein of semen coagulum, inhibits human sperm capacitation by interfering with the superoxide anion generated during this process. J Androl 2001;22:672–679. [PubMed] [Google Scholar]
- 54. Gonzales GF, Kortebani G, Mazzolli AB. Hyperviscosity and hypofunction of the seminal vesicles. Arch Androl 1993;30:63–68. [DOI] [PubMed] [Google Scholar]
- 55. Beer R, Weisselberg M, Soffer Y, Lewin LM. Human seminal fluid γ‐glutamyl‐transferase: a prostatic marker enzyme. Int J Androl 1978;1:405–410. [Google Scholar]
- 56. von der Kammer H, Scheit KH, Weidner W, Cooper TG. The evaluation of markers of prostatic function. Urol Res 1991;19:343–347. [DOI] [PubMed] [Google Scholar]
- 57. Comhaire F, Vermeulen L, Pieters O. Study of the accuracy of physical and biochemical markers in semen to detect infectious dysfunction of the accessory sex glands. Int J Androl 1989;10:50–53. [DOI] [PubMed] [Google Scholar]
- 58. Sorensen MB, Bergdhal IA, Hjollund NH, Bonde JP, Stoltenberg M, Ernst E. Zinc, magnesium and calcium in human seminal fluid: relations to other semen parameters and fertility. Mol Hum Reprod 1999;5:331–337. [DOI] [PubMed] [Google Scholar]
- 59. Dieudonne O, Godin PA, Van‐Langendonckt A, Jamart J, Galanti L. Biochemical analysis of the sperm and infertility. Clin Chem Lab Med 2001;39:455–457. [DOI] [PubMed] [Google Scholar]
- 60. Caldamone AA, Emilson LBV, Al‐Juburi A, Cockett ATK. Prostatitis: prostatic secretory dysfunction affecting fertility. Fertil Steril 1980;34:602–603. [PubMed] [Google Scholar]
- 61. Polakoski KL, Syner FN, Zaneveld LJD. Biochemistry of the human seminal plasma In: Hafez ESE, editor. Human semen and fertility regulation in men. Saint Louis: The C.V. Mosby Company; 1976. p 133–143. [Google Scholar]
- 62. Wolff H, Bezold G, Zebhauser M, Meurer M. Impact of clinically silent inflammation on male genital tract organs as reflected by biochemical markers in semen. J Androl 1991;12:331–334. [PubMed] [Google Scholar]
- 63. Wolff H, Anderson DJ. Immunohistologic characterization and quantitation of leukocyte subpopulations in human semen. Fertil Steril 1988;49:497–504. [PubMed] [Google Scholar]
- 64. Zorn B, Virant‐Klun I, Meden‐Vrtovec H. Semen granulocyte elastase: its relevance for the diagnosis and prognosis of silent genital tract inflammation. Hum Reprod 2000;15:1978–1984. [DOI] [PubMed] [Google Scholar]
- 65. Wolff H. The biologic significance of white blood cells in semen. Fertil Steril 1995;63:1143–1157. [DOI] [PubMed] [Google Scholar]
- 66. Aitken RJ, Baker HWG. Seminal leukocytes: passengers, terrorists or good samaritans? Hum Reprod 1995;10:1736–1739. [DOI] [PubMed] [Google Scholar]
- 67. Barratt CL, Bolton AE, Cooke ID. Functional significance of white blood cells in the male and female reproductive tract. Hum Reprod 1990;5:639–648. [DOI] [PubMed] [Google Scholar]
- 68. Bar‐Chama N, Goluboff E, Fisch H. Infection and pyospermia in male infertility,,, Is it really a problem? Urol Clin North Am 1994;21:469–475. [PubMed] [Google Scholar]
- 69. Drevius LO, Ericksson H. Osmotic swelling of mammalian spermatozoa. Exp Cell Res 1966;42:136–156. [DOI] [PubMed] [Google Scholar]
- 70. Jeyendran RS, Van der Ven HH, Perez‐Pelaez M, Crabo BG, Zaneveld LJD. Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics. J Reprod Fertil 1984;70:219–228. [DOI] [PubMed] [Google Scholar]
- 71. Takahashi K, Uchida A, Kitao M. Hypoosmotic swelling test of sperm. Arch Androl 1990;25:225–242. [DOI] [PubMed] [Google Scholar]
- 72. Naz RK, Menge AC. Antisperm antibodies: origin, regulation, and sperm reactivity in human infertility. Fertil Steril 1994;61:1001–1013. [DOI] [PubMed] [Google Scholar]
- 73. Mazumdar S, Levine AS. Antisperm antibodies: etiology, pathogenesis, diagnosis, and treatment. Fertil Steril 1998;70:799–810. [DOI] [PubMed] [Google Scholar]
- 74. Lombardo F, Gandini L, Dondero F, Lenzi A. Immunology and immunopathology of the male genital tract. Antisperm immunity in natural and assisted reproduction. Hum Reprod Update 2001;7:450–456. [DOI] [PubMed] [Google Scholar]
- 75. de Lamirande E, Gagnon C. Impact of reactive oxygen species on spermatozoa: a balancing act between beneficial and detrimental effects. Hum Reprod 1995;10(suppl 1)15–21. [DOI] [PubMed] [Google Scholar]
- 76. Saleh RA, Agarwal A. Oxidative stress and male infertility: from research bench to clinical practice. J Androl 2002;23:737–752. [PubMed] [Google Scholar]
- 77. Aitken RJ. The Amoroso lecture. The human spermatozoon—a cell in crisis? J Reprod Fertil 1999;115:1–7. [DOI] [PubMed] [Google Scholar]
- 78. Ochsendorf FR. Infections in the male genital tract and reactive oxygen species. Hum Reprod Update 1999;5:399–420. [DOI] [PubMed] [Google Scholar]
- 79. Ollero M, Gil‐Guzman E, Lopez MC, et al. Characterization of subsets of human spermatozoa at different stages of maturation: implications in the diagnosis and treatment of male infertility. Hum Reprod 2001;16:1912–1921. [DOI] [PubMed] [Google Scholar]
- 80. Sakkas D, Mariethoz E, Manicardi G, Bizzaro D, Bianchi PG, Bianchi U. Origin of DNA damage in ejaculated human spermatozoa. Rev Reprod 1999;4:31–37. [DOI] [PubMed] [Google Scholar]
- 81. Evenson DP, Larson KL, Jost LK. Sperm chromatin structure assay: its clinical use for detecting sperm DNA fragmentation in male infertility and comparisons with other techniques. J Androl 2002;23:25–43. [DOI] [PubMed] [Google Scholar]
- 82. Seligman J, Shalgi R, Oschry Y, Kosower NS. Sperm analysis by flow cytometry using the fluorescent thiol labeling agent monobromobimane. Mol Reprod Dev 1991;29:276–281. [DOI] [PubMed] [Google Scholar]
- 83. Evenson DP, Darzynkiewicz Z, Melamed MR. Relation of mammalian sperm chromatin heterogeneity to fertility. Science 1980;210:1131–1133. [DOI] [PubMed] [Google Scholar]
- 84. Terquem A, Dadoune JP. Aniline blue staining of human spermatozoon chromatin. Evaluation of nuclear maturation In: Andre J, ed. The sperm cell. The Hague: Martinus Nijhoff Publishers; 1980. p 249–252. [Google Scholar]
- 85. Tejada RI, Mitchell JC, Norman A, Marik JJ, Friedman S. A test for the practical evaluation of male fertility by acridine orange (AO) fluorescence. Fertil Steril 1984;42:87–91. [DOI] [PubMed] [Google Scholar]
- 86. Sailer BL, Jost LK, Evenson DP. Mammalian sperm DNA susceptibility to in situ denaturation associated with the presence of DNA strand breaks as measured by the terminal deoxynucleotidyl transferase assay. J Androl 1995;16:80–87. [PubMed] [Google Scholar]
- 87. Hughes CM, Lewis SE, McKelvey‐Martin VJ, Thompson W. A comparison of baseline and induced DNA damage in human spermatozoa from fertile and infertile men, using a modified comet assay. Mol Hum Reprod 1996;2:613–619. [DOI] [PubMed] [Google Scholar]
- 88. Manicardi GC, Tombacco A, Bizzaro D, Bianchi U, Bianchi PG, Sakkas D. DNA strand breaks in ejaculated human spermatozoa: comparison of susceptibility to the nick translation and terminal transferase assays. Histochem J 1998;30:33–39. [DOI] [PubMed] [Google Scholar]
- 89. Mortimer ST. CASA—Practical aspects. J Androl 2000;21:515–524. [PubMed] [Google Scholar]
- 90. Suarez SS. Hyperactivated motility in sperm. J Androl 1996;17:331–335. [PubMed] [Google Scholar]
- 91. Schill WB. Some disturbances of acrosomal development and function in human spermatozoa. Hum Reprod 1991;6:969–978. [DOI] [PubMed] [Google Scholar]
- 92. Baccetti B, Burrini AG, Capitani S, et al. Notulae seminologicae. 2. The ‘short tail’ and ‘stump’ defect in human spermatozoa. Andrologia 1993;25:331–335. [PubMed] [Google Scholar]
- 93. Viville S, Mollard R, Bach ML, Falquet C, Gerlinger P, Warter S. Do morphological anomalies reflect chromosomal aneuploidies?: case report. Hum Reprod 2000;15:2563–2566. [DOI] [PubMed] [Google Scholar]
- 94. Kalahanis J, Rousso D, Kourtis A, Mavromatidis G, Makedos G, Panidis D. Round‐headed spermatozoa in semen specimens from fertile and subfertile men. J Reprod Med 2002;47:489–493. [PubMed] [Google Scholar]
