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Chapter 3 - Basic Semen Examination

Published online by Cambridge University Press:  16 February 2022

David Mortimer ,
Lars Björndahl ,
Christopher L. R. Barratt ,
José Antonio Castilla ,
Roelof Menkveld ,
Ulrik Kvist ,
Juan G. Alvarez  and
Trine B. Haugen
David Mortimer
Affiliation:
Oozoa Biomedical Inc., Vancouver
Lars Björndahl
Affiliation:
Karolinska Institutet, Stockholm
Christopher L. R. Barratt
Affiliation:
University of Dundee
José Antonio Castilla
Affiliation:
HU Virgen de las Nieves, Granada
Roelof Menkveld
Affiliation:
Stellenbosch University, South Africa
Ulrik Kvist
Affiliation:
Karolinska Institutet, Stockholm
Juan G. Alvarez
Affiliation:
Centro ANDROGEN, La Coruña
Trine B. Haugen
Affiliation:
Oslo Metropolitan University
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Summary

Provides reference methods for all aspects of a basic semen examination that are structured as standard operating procedures (SOPs) for easy use at the bench. Methods are focussed on objectivity, robustness, standardized reporting, controlling the risk of errors, and minimizing measurement uncertainty. Includes ejaculate physical characteristics, volume and pH, sperm concentration, sperm motility, sperm vitality, sperm morphology (including seminal cytology), and the simple detection of peroxidase-positive leukocytes in semen.

Keywords

semen analysisspermatozoasperm countsperm concentrationsperm motilitysperm morpholgysperm vitality
Type
Chapter
Information
A Practical Guide to Basic Laboratory Andrology , pp. 34 - 80
Publisher: Cambridge University Press
Print publication year: 2022

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References

Kvist, U, Björndahl, L, eds. Manual on Basic Semen Analysis. Oxford: Oxford University Press, 2002.Google Scholar
Mortimer, D. Practical Laboratory Andrology. Oxford: Oxford University Press, 1994.CrossRefGoogle Scholar
Pound, N, Javed, MH, Ruberto, C, et al. Duration of sexual arousal predicts semen parameters for masturbatory ejaculates. Physiol Behav 2002; 76: 685–9.CrossRefGoogle ScholarPubMed
Zavos, PM, Goodpasture, JC. Clinical improvements of specific seminal deficiencies via intercourse with a seminal collection device versus masturbation. Fertil Steril 1989; 51: 190–3.Google Scholar
Kamischke, A, Nieschlag, E. Treatment of retrograde ejaculation and anejaculation. Hum Reprod Update 1999; 5: 448–74.CrossRefGoogle ScholarPubMed
MacLeod, J, Gold, RZ. The male factor in fertility and infertility. V. Effect of continence on semen quality. Fertil Steril 1952; 3: 297–315.CrossRefGoogle ScholarPubMed
Carlsen, E, Andersson, AM, Petersen, JH, Skakkebaek, NE. History of febrile illness and variation in semen quality. Hum Reprod 2003; 18: 2089–92.CrossRefGoogle ScholarPubMed
Cooper, TG, Brazil, C, Swan, SH, Overstreet, JW. Ejaculate volume is seriously underestimated when semen is pipetted or decanted into cylinders from the collection vessel. J Androl 2007; 28: 1–4.CrossRefGoogle ScholarPubMed
Björndahl, L, Kvist, U. Sequence of ejaculation affects the spermatozoon as a carrier and its message. Reprod Biomed Online 2003; 7: 440–8.CrossRefGoogle ScholarPubMed
MacLeod, J, Gold, RZ. The male factor in fertility and infertility. III. An analysis of motile activity in the spermatozoa of 1000 fertile men and 1000 men in infertile marriage. Fertil Steril 1951; 2: 187–204.CrossRefGoogle ScholarPubMed
World Health Organization. WHO Laboratory Manual for the Examination of Human Semen and Sperm-Cervical Mucus Interactions, 4th edn. Cambridge: Cambridge University Press, 1999.Google Scholar
World Health Organization. WHO Laboratory Manual for the Examination and Processing of Human Semen, 6th edn. Geneva: World Health Organization, 2021.Google Scholar
International Organization for Standardization. ISO 23162:2021 Basic Semen Examination – Specification and Test Methods. Geneva: International Organization for Standardization, 2021.Google Scholar
Mortimer, D. Laboratory standards in routine clinical andrology. Reprod Med Review 1994; 3: 97–111.Google Scholar
Sifer, C, Sasportes, T, Barraud, V, et al. World Health Organization grade ‘a’ motility and zona-binding test accurately predict IVF outcome for mild male factor and unexplained infertilities. Hum Reprod 2005; 20: 2769–75.Google Scholar
Verheyen, G, Tournaye, H, Staessen, C, et al. Controlled comparison of conventional in-vitro fertilization and intracytoplasmic sperm injection in patients with asthenozoospermia. Hum Reprod 1999; 14: 2313–19.CrossRefGoogle ScholarPubMed
Barratt, CLR, Björndahl, L, Menkveld, R, Mortimer, D. The ESHRE Special Interest Group for Andrology Basic Semen Analysis Course: a continued focus on accuracy, quality, efficiency and clinical relevance. Hum Reprod 2011; 26: 3207–12.CrossRefGoogle ScholarPubMed
Mortimer, D, Shu, MA, Tan, R. Standardization and quality control of sperm concentration and sperm motility counts in semen analysis. Hum Reprod 1986; 1: 299–303.CrossRefGoogle ScholarPubMed
Mortimer, D, Shu, MA, Tan, R, Mortimer, ST. A technical note on diluting semen for the haemocytometric determination of sperm concentration. Hum Reprod 1989; 4: 166–8.Google Scholar
Kirkman Brown, J, Björndahl, L. Evaluation of a disposable plastic Neubauer counting chamber for semen analysis. Fertil Steril 2009; 91: 627–31.CrossRefGoogle ScholarPubMed
Zuvela, E, Matson, P. Performance of four chambers to measure sperm concentration: results from an external quality assurance programme. Reprod Biomed Online 2020; 41: 671–8.CrossRefGoogle ScholarPubMed
Mortimer, D. A technical note on the assessment of human sperm vitality using eosin-nigrosin staining. Reprod Biomed Online 2020; 40: 851–5.CrossRefGoogle ScholarPubMed
Björndahl, L, Söderlund, I, Johansson, S, et al. Why the WHO recommendations for eosin-nigrosin staining techniques for human sperm vitality assessment must change. J Androl 2004; 25: 671–8.CrossRefGoogle ScholarPubMed
Björndahl, L, Söderlund, I, Kvist, U. Evaluation of the one-step eosin-nigrosin staining technique for human sperm vitality assessment. Hum Reprod 2003; 18: 813–16.Google Scholar
Hühner, M. Methods of examining for spermatozoa in the diagnosis and treatment of sterility. Int J Surg 1921; 34: 91–100.Google Scholar
Cary, WH. Sterility diagnosis: The study of sperm cell migration in female secretion and interpretation of findings. NY State J Med 1930; 30: 131–6.Google Scholar
Cary, WH, Hotchkiss, RS. Semen appraisal. A differential stain that advances the study of cell morphology. JAMA 1934; 102: 587–90.Google Scholar
Moench, GL, Holt, H. Biometrical studies of head lengths of human spermatozoa. J Lab Clin Med 1932; 17: 297–316.Google Scholar
Menkveld, R, Van Zyl, JA, Kotze, T, Joubert, G. Possible changes in male fertility over a 15-year period. Arch Androl 1986; 17: 143–4.Google Scholar
Mortimer, D. Selectivity of sperm transport in the female genital tract. In: Cohen, J, Hendry, WF, eds. Spermatozoa, Antibodies and Infertility. Oxford & London: Blackwell Scientific Publications, 1978.Google Scholar
Mortimer, D. Sperm form and function: Beauty is in the eye of the beholder. In: van der Horst, G, Franken, D, Bornman, R, et al., eds. Proceedings of the 9th International Symposium on Spermatology. Bologna: Monduzzi Editore, 2002.Google Scholar
Holt, WV, Fazeli, A. Do sperm possess a molecular passport? Mechanistic insights into sperm selection in the female reproductive tract. Mol Hum Reprod 2015; 21: 491–501.Google Scholar
Kölle, S. Transport, distribution and elimination of mammalian sperm following natural mating and insemination. Reprod Dom Anim 2015; 50: 2–6.Google Scholar
Miller, DJ. The epic journey of sperm through the female reproductive tract. Animal 2018; 12 Suppl 1: S110–20.Google Scholar
Shojaei Saadi, HA, van Riemsdijk, E, Dance, AL, et al. Proteins associated with critical sperm functions and sperm head shape are differentially expressed in morphologically abnormal bovine sperm induced by scrotal insulation. J Proteomics 2013; 82: 64–80.CrossRefGoogle ScholarPubMed
Menkveld, R. An investigation of environmental influences on spermatogenesis and semen parameters. PhD Dissertation, Faculty of Medicine, University of Stellenbosch, South Africa, 1987.Google Scholar
Menkveld, R, Stander, FS, Kotze, TJ, et al. The evaluation of morphological characteristics of human spermatozoa according to stricter criteria. Hum Reprod 1990; 5: 586–92.Google Scholar
Coetzee, K, Kruge, TF, Lombard, CJ. Predictive value of normal sperm morphology: a structured literature review. Hum Reprod Update 1998; 4: 73–82.Google Scholar
Menkveld, R. The basic semen analysis. In: Oehninger, S, Kruger, TF, eds. Male Infertility. London: Informa Healthcare, 2007, 141–70.Google Scholar
Papanicolaou, GN. A new procedure for staining vaginal smears. Science 1942; 95: 438–9.CrossRefGoogle ScholarPubMed
Oettlé, EE. Using a new acrosome stain to evaluate sperm morphology. Vet Med 1986; 81: 263–6.Google Scholar
Henkel, R, Schreiber, G, Sturmhoefel, A, et al. Comparison of three staining methods for the morphological evaluation of human spermatozoa. Fertil Steril 2008; 89: 449–55.CrossRefGoogle ScholarPubMed
van der Horst, G, Maree, L. SpermBlue: a new universal stain for human and animal sperm which is also amenable to automated sperm morphology analysis. Biotech Histochem 2009; 84: 299–308.Google Scholar
Maree, L, du Plessis, SS, Menkveld, R, van der Horst, G. Morphometric dimensions of the human sperm head depend on the staining method used. Hum Reprod 2010; 25: 1369–82.Google Scholar
World Health Organization. WHO Laboratory Manual for the Examination of Human Semen and Sperm-Cervical Mucus Interactions, 3rd edn. Cambridge: Cambridge University Press, 1992.Google Scholar
Mortimer, D, Menkveld, R. Sperm morphology assessment – Historical perspectives and current opinions. J Androl 2001; 22: 192–205.CrossRefGoogle ScholarPubMed
Menkveld, R. Clinical significance of the low normal sperm morphology value as proposed in the fifth edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen. Asian J Androl 2010; 12: 47–58.Google Scholar
Mortimer, D. The functional anatomy of the human spermatozoon: relating ultrastructure and function. Mol Hum Reprod 2018; 24: 567–92.Google ScholarPubMed
Oettlé, EE, Menkveld, R, Swanson, RJ. Photographs with interpretations. In: Menkveld, R, Oettlé, EE, Kruger, TF, et al., eds. Atlas of Human Sperm Morphology. Baltimore: Williams & Wilkins, 1991, 15–65.Google Scholar
Cooper, TG, Yeung, CH, Fetic, S, et al. Cytoplasmic droplets are normal structures of human sperm but are not well preserved by routine procedures for assessing sperm morphology. Hum Reprod 2004; 19: 2283–8.Google Scholar
World Health Organization. WHO Laboratory Manual for the Examination and Processing of Human Semen, 5th edn. Geneva: World Health Organization, 2010.Google Scholar
Kruger, TF, Menkveld, R, Stander, FSH, et al. Sperm morphological features as a prognostic factor in in vitro fertilization. Fertil Steril 1986; 46: 1118–23.CrossRefGoogle ScholarPubMed
Endtz, AW. A rapid staining method for differentiating granulocytes from ‘germinal cells’ in Papanicolaou-stained semen. Acta Cytol 1974; 18: 2–7.Google Scholar
Wolff, H. The biologic significance of white blood cells in semen. Fertil Steril 1995; 63: 1143–57.Google Scholar
Lackner, J, Schatzl, G, Horvath, S, et al. Value of counting white blood cells (WBC) in semen samples to predict the presence of bacteria. Eur Urol 2006; 49: 148–52; discussion 52–3.Google Scholar
Punab, M, Loivukene, K, Kermes, K, Mandar, R. The limit of leucocytospermia from the microbiological viewpoint. Andrologia 2003; 35: 271–8.CrossRefGoogle ScholarPubMed
Rodin, DM, Larone, D, Goldstein, M. Relationship between semen cultures, leukospermia, and semen analysis in men undergoing fertility evaluation. Fertil Steril 2003; 79 Suppl 3: 1555–8.CrossRefGoogle ScholarPubMed
Bedford, JM, Zelikovsky, G. Viability of spermatozoa in the human ejaculate after vasectomy. Fertil Steril 1979; 32: 460–3.CrossRefGoogle ScholarPubMed
Lewis, EL, Brazil, CK, Overstreet, JW. Human sperm function in the ejaculate following vasectomy. Fertil Steril 1984; 42: 895–8.Google Scholar
Jouannet, P, David, G. Evolution of the properties of semen immediately following vasectomy. Fertil Steril 1978; 29: 435–41.Google Scholar
Hancock, P, Woodward, BJ, Muneer, A, Kirkman-Brown, JC. 2016 Laboratory guidelines for postvasectomy semen analysis: Association of Biomedical Andrologists, the British Andrology Society and the British Association of Urological Surgeons. J Clin Pathol 2016; 69: 655–60.CrossRefGoogle Scholar
Breitinger, MC, Roszkowski, EH, Bauermeister, AJ, Rosenthal, AA. Duplicate vas deferens encountered during inguinal hernia repair: a case report and literature review. Case Rep Surg 2016; 2016: 8324925.Google Scholar
Cooper, TG, Yeung, CH, Nashan, D, Jockenhovel, F, Nieschlag, E. Improvement in the assessment of human epididymal function by the use of inhibitors in the assay of alpha-glucosidase in seminal plasma. Int J Androl 1990; 13: 297–305.Google Scholar

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