Andrology laboratory review: evaluation of sperm morphology
Keywords:
Sperm, morphology, fertility, animal
Abstract
Sperm morphology assessment has an important role in male fertility diagnosis and prognosis, both for humans and animals. Thus, it is important that relevant results are comparable and consistent. To achieve these aims, the following procedures are recommended: a. semen sample is suitably ‘fixed’ (e.g. in isotonic buffered formal-saline); b. sperm are examined at 1,000 x (phase or DIC microscopy); c.at least 200 sperm are counted; d. each sperm is placed into 1 category, only (e.g. normal, head, midpiece etc), and e. 70% ‘normal’ sperm is the threshold for a satisfactory sample. In addition, morphologists should be provided with relevant continuing education, upskilling, and monitoring programs. This review provides guidelines for the best performance of this assessment, as well as for avoiding pitfalls.
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References
1. Brito LFC, Althouse GA, Aurich C, et al: Andrology laboratory review: evaluation of sperm concentration. Theriogenology 2016;85:1507-1527. doi: 10.1016/j.theriogenology.2016.01.002
2. Peter AT, Brito LF, Althouse GC, et al: Andrology laboratory review: evaluation of sperm motility. Clinical Theriogenology 2021;13:297-315. doi: 10.58292/ct.v13.9359
3. Mortimer D, Menkveld R: Sperm morphology assessment: historical perspectives and current opinions. J Androl 2001;22:192-205. doi: 10.1002/j.1939-4640.2001.tb02171.x
4. Rodriguez-Martinez H: Semen evaluation and handling: emerging techniques and future development. In: Chenoweth PJ, Lorton SP: editors. Animal Andrology. Theories and Applications. Wallingford; CABI: 2014. p. 509-549.
5. Sathe S, Shipley CF: Applied andrology in sheep, goats and selected cervids. In: Chenoweth PJ, Lorton SP: editors. Animal Andrology. Theories and Applications. Wallingford; CABI: 2014. p. 226-253.
6. Mortimer D: The functional anatomy of the human spermatozoon; relating ultrastructure and function. Mol Hum Reprod 2018;24:567-592. doi: 10.1093/molehr/gay040
7. Encisco M, Cisale H, Johnston SD, et al: Major morphological sperm abnormalities in the bull are related to sperm DNA damage. Theriogenology 2011;76:23-32. doi: 10.1016/j.theriogenology.2010.12.034
8. Erenpreiss J, Spano M, Erenpreisa J, et al: Sperm chromatin structure and male fertility: biological and clinical aspects. Asian J Androl 2006;8:11-29. doi: 10.1111/j.1745-7262.2006.00112.x
9. Brito LF: Evaluation of stallion sperm morphology. Clin Tech Equine Pract 2007;6:249-264. doi: 10.1053/j.ctep.2007.09.004
10. 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. doi: 10.3109/10520290902984274
11. Chang V, Garcia A, Hitschfield N, et al: Gold-standard for computer-assisted morphological sperm analysis. Comput Biol Med 2017;83:43-150. doi: 10.1016/j.compbiomed.2017.03.004
12. Reeves A, Klabnik J, Strickland L, et al: Bull sperm morphology assessment varied by evaluator. Clinical Theriogenology 2022;14:11-17. doi: 10.58292/ct.v14.9293
13. Mortimer D: Practical Laboratory Andrology. Oxford; Oxford University Press: 1994. p. 393.
14. Chenoweth PJ: Sperm morphology. In: Chenoweth PJ, Lorton SP: editors. Manual of Animal Andrology. CABI: Wallingford; 2022. p. 45-55.
15. Hackett AJ, MacPherson JW: Some staining procedures for spermatozoa. A review. Can Vet J 1965;6:55-62.
16. Amann RP, Waberski D: Computer-assisted sperm analysis (CASA): capabilities and potential developments. Theriogenology 2014;81:5-17. doi: 10.1016/j.theriogenology.2013.09.004
17. Mortimer S, van der Horst G, Mortimer D: The future of computer-aided sperm analysis. Asian J Androl 2015;17:545-553. doi: 10.4103/1008-682X.154312
18. Keel BA, Stembridge TW, Pineda G, et al: Lack of standardization in performance of the semen analysis among laboratories in the United States. Fertil Steril 2002;783;603-608. doi: 10.1016/S0015-0282(02)03296-X
19. Moretti E, Sutera G, Collodel G: The importance of transmission electron microscopy analysis of spermatozoa: Diagnostic applications and basic research. Syst Biol Reprod Med 2016;62:171-183. doi: 10.3109/19396368.2016.1155242
20. Soley JT, du Plessis L: Ultra-imaging in applied animal andrology: the power and the beauty. Anim Reprod Sci 2020;220:106306. doi: 10.1016/j.anireprosci.2020.106306
21. Hancock JL, Trevan DJ: The acrosome and post-nuclear cap of bull spermatozoa. Royal J Microscop Soc 1957;76:77-83. doi: 10.1111/j.1365-2818.1956.tb00442.x
22. Kuster CE, Singer RS, Althouse GC: Determining sample size for the morphological assessment of sperm. Theriogenology 2004;61:691-703. doi: 10.1016/S0093-691X(03)00240-1
23. Mortimer D: A technical note on the assessment of human sperm vitality using eosin-nigrosin staining. Reprod Biomed Online 2020;40:851-855. doi: 10.1016/j.rbmo.2020.03.002
24. WHO laboratory manual for the examination and processing of human semen. 6th edition, WHO Press: 2021.
25. Kaya A, Birler S, Enwall L, et al: Determinants of sperm morphology. In: Chenoweth PJ, Lorton SP: editors. Manual of Animal Andrology. CABI; Wallingford: 2014. p. 34-56.
26. Blom E: Interpretation of spermatic cytology in bulls. Fertil Steril 1950;1:223-238. doi: 10.1016/S0015-0282(16)30183-2
27. Blom E: The ultrastructure of some characteristic sperm defects and a proposal for a new classification of the bull spermiogram. Atti del VII Simposia Int di Zootecnia. Milan 1972;7:125-139.
28. Rao AR: Changes in the morphology of sperm during their passage through the genital tract of bulls with normal and impaired spermatogenesis. PhD Thesis Royal Vet College, Stockholm: 1971.
29. Bjorndahl L, Mortimer D, Barratt CLR, et al: A Practical Guide to Basic Laboratory Andrology. Cambridge; Cambridge University Press: 2010. p. 336.
30. Saacke RG: Sperm morphology: its relevance to compensable and uncompensable traits in semen. Theriogenology 2008;70:473-478. doi: 10.1016/j.theriogenology.2008.04.012
31. Chemes HE, Rawe YV: Sperm pathology: a step beyond descriptive morphology. Origin, characterization and fertility potential of abnormal sperm phenotypes in infertile men. Hum Reprod Update 2003;9:405-428. doi: 10.1093/humupd/dmg034
32. Chenoweth PJ, McPherson FJ: Genetic aspects of male reproduction. In: Chenoweth PJ, Lorton SP: editors. Animal Andrology: Theories and Applications. CABI; Wallingford: 2014. p. 144-173.
33. Chenoweth PJ: Genetic sperm defects. Theriogenology 2005;64:457-468. doi: 10.1016/j.theriogenology.2005.05.005
34. Lorton SP: Evaluation of semen in the andrology laboratory. In: Chenoweth PJ, Lorton SP: editors. Animal Andrology. Theories and Applications. CABI; Wallingford: 2014. p. 100-143.
35. Cairo Consensus Workshop Group. The current status and future of andrology: a consensus report from the Cairo workshop group. Andrology 2020;8:26–52. doi: 10.1111/andr.12720
36. Brito LF, Greene LM, Kelleman, et al: Effect of method and clinician on stallion sperm morphology evaluation. Theriogenology 2011;76:745-750. doi: 10.1016/j.theriogenology.2011.04.007
37. Frenaeu GE, Chenoweth PJ, Ellis R, et al: Sperm morphology of beef bulls evaluated by two different methods. Anim Reprod Sci 2010;118:176-181. doi: 10.1016/j.anireprosci.2009.08.015
38. Menon AG, Thundathil JC, Wilde R, et al: Validating the assessment of bull sperm morphology by veterinary practitioners. Can Vet J 2011;52:407-408.
39. Chacón J: Assessment of sperm morphology in Zebu bulls, under field conditions in the tropics. Reprod Dom Anim 2001;36:91-107. doi: 10.1046/j.1439-0531.2001.00253.x
40. Cooper TG, Atkinson AD, Nieschlag E: Experience with external quality control in spermatology. Hum Reprod 1999;3:765-769. doi: 10.1093/humrep/14.3.765
41. Alvarez C, Castilla JA, Ramírez JP, et al: External quality control program for semen analysis: Spanish experience. J Assist Reprod Genet 2005;22:379-387. doi: 10.1007/s10815-005-7461-2
42. Wiltbank JN and Parish NR: Pregnancy rate in cows and heifers bred to bulls selected for semen quality. Theriogenology 1986;25:779-783. doi: 10.1016/0093-691X(86)90093-2
43. McPherson FJ, Nielsen SG, Chenoweth PJ: Semen effects on insemination outcomes in sows. Anim Reprod Sci 2014;151:28-33. doi: 10.1016/j.anireprosci.2014.09.021
44. Pukazhenthi BS, Wildt DE, Howard JG: The phenomenon and significance of teratospermia in felids. J Reprod Fertil Suppl 2001;57:423-433.
45. Waberski D, Riesenbeck A, Schulze M, et al: Application of preserved boar semen for artificial insemination: past, present and future challenges. Theriogenology 2019;137:2-7. doi: 10.1016/j.theriogenology.2019.05.030
46. Clark G: Staining Procedures. Biological Stain Commission. 4th edition, Baltimore; Williams and Wilkins: 1981.
47. Blom E: A one-minute live-dead stain by means of eosin nigrosin. Fertil Steril 1950;1:176-177. doi: 10.1016/S0015-0282(16)30125-X
48. Bjorndahl L, Soderlund I, Kvist U: Evaluation of the one-step eosin-nigrosin staining technique for human sperm vitality assessment. Hum Reprod 2003;18:813-816. doi: 10.1093/humrep/deg199
49. Saacke RG, Marshall CE: Observations on the acrosomal cap of fixed and unfixed bovine spermatozoa. J Reprod Fertil 1968;16:511-514. doi: 10.1530/jrf.0.0160511
50. Johnson L, Berndtsen WE, Pickett BW: An improved method for evaluating acrosomes of bovine spermatozoa. J Anim Sci 1976;42:951-954. doi: 10.2527/jas1976.424951x
51. Hanson R., Reddick S., Thuerauf S, et al: Comparison of bull sperm morphology evaluation methods under field conditions. Clinical Therigenology 2023;15:1-9. doi: 10.58292/CT.v15.9425
52. Morselli MG, Colombo M, Faustini M, et al: Morphological indices for canine spermatozoa based on the World Health Organization laboratory manual for human semen. Reprod Domest Anim 2019;54:949-955. doi: 10.1111/rda.13440
53. Zhu WJ: Preparation and observation methods can produce mispleading artefacts in human sperm ultrastructural morphology. Andrologia 2018;50:e13043. doi: 10.1111/and.13043
54. O’Connell M, McClure N, Lewis SEM: The effects of cryopreservation on sperm morphology, motility and mitochondrial function. Hum Reprod 2002;17:704-709. doi: 10.1093/humrep/17.3.704
55. Spindler R, Keeley T, Satake N: Applied andrology in endangered, exotic and wildlife species. In: Chenoweth PJ, Lorton SP: editors. Animal Andrology. Theories and Applications. CABI; Wallingford: 2014. p. 450-473.
56. Dey S, Kharbuli SM, Chakraborty R, et al: Effect of environmental acid-stress on the sperm of a hill-stream fish devario aequipinnatus: a scanning electron microscopic evaluation. Microsc Res Tech 2009;72:76-78. doi: 10.1002/jemt.20640
57. Erenpreiss J, Hlevicka S, Zalkalns J, et al: Effect of Leukocytospermia on Sperm DNA integrity: a negative effect in abnormal semen samples. J Androl 2002;23:717-723. doi: 10.1002/j.1939-4640.2002.tb02315.x
58. Althouse GC, Lu K: Bacteriospermia in extended porcine semen. Theriogenology 2005;63:573-584. doi: 10.1016/j.theriogenology.2004.09.031
59. Moretti E, Capitani S, Figura N, et al: The presence of bacteria species in semen and sperm quality. J Assist Reprod Genet 2009;26;47-56. doi: 10.1007/s10815-008-9283-5
60. Dowsett KF, Knott LM: The influence of age and breed on stallion semen. Theriogenology 1996;46:397-412. doi: 10.1016/0093-691X(96)00162-8
61. Rijsselaere T, Maes D, Hoflack G, et al: Effect of body weight, age and breeding history on canine sperm quality parameters measured by the Hamilton-Thorne analyser. Reprod Domest Anim 2007;42:143-148. doi: 10.1111/j.1439-0531.2006.00743.x
62. Hesser A, Darr C, Gonzales K, et al: Semen evaluation and fertility assessment in a purebred dog breeding facility. Theriogenology 2017;87:115-123. doi: 10.1016/j.theriogenology.2016.08.012
63. Johananison E, Campana A, Luthi R, et al: Evaluation of ‘round cells’ in semen analysis: a comparative study. Hum Reprod Update 2000;6:404-412. doi: 10.1093/humupd/6.4.404
64. Aitken RJ, Baker HWG: Seminal leukocytes: passengers, terrorists or good Samaritans. Hum Reprod 1995;10:1736-1739. doi: 10.1093/oxfordjournals.humrep.a136165
65. González-Marín G, Roy R, López-Fernández C, et al: Bacteria in bovine semen can increase sperm DNA fragmentation rates: a kinetic experimental approach. Anim Reprod Sci 2011;123:139-148. doi: 10.1016/j.anireprosci.2010.11.014
66. Keel BA, Webster BW: Handbook of the laboratory diagnosis and treatment of infertility. Boca Raton; CRC Press: 2000. p. 431.
67. Watson PF: Use of a Giemsa stain to detect changes in the acrosome of frozen ram semen. Vet Rec 1975;97:12-15. doi: 10.1136/vr.97.1.12
68. Williams WW, Savage A: Observations upon the seminal micropathology of bulls. Cornell Vet 1925;15:353-375.
69. Lagerlóf N: Changes in the spermatozoa and in the testes of bulls with impaired or enhanced fertility. Acta Pathol Microbiol Scand 1934;19: p. 254.
70. Oettle EE: Using a new acrosome stain to evaluate sperm morphology. Vet Med 1986;81:263-266.
71. Chan PJ, Corselli JU, Jacobson WC, et al: Spermac stain analysis of human sperm acrosomes. Fertil Steril 1999;72:124-128. doi: 10.1016/s0015-0282(99)00201-0
72. Sousa APM, Tavares RS, de la Calle JFV, et al: Dual use of Diff-Quik-like stains for the simultaneous evaluation of human sperm morphology and chromatin status. Hum Reprod 2009;24:28-36. doi: 10.1093/humrep/den365
73. Pozor MA, Zambrano GL, Runcin E, et al: Usefulness of dip quick stain in evaluating sperm morphology in stallions. Proc Am Assoc Equine Pract 2012;58:506-510.
74. Barth AD, Oko RJ: Abnormal Morphology of Bovine Spermatozoa. 1st edition, Ames, IA; Iowa State University Press: 1989. p. 302.
75. Erenpreiss J, Jepson K, Giwercman A, et al: Toluidine blue cytometry test for sperm DNA conformation: comparison with the flow cytometric sperm chromatin structure and TUNEL assays. Hum Reprod 2004;19:2277-2282. doi: 10.1093/humrep/deh417
76. Tsarev I, Bungum M, Giwercman A, et al: Evaluation of male fertility potential by Toludine Blue test for sperm chromatin structural assessment. Hum Reprod 2009;24:1569-1574. doi: 10.1093/humrep/dep068
77. Kovacs A, Foote RH: Viability and acrosome staining of bull, boar and rabbit spermatozoa. Biotech Histochem 1992;67:119-124. doi: 10.3109/10520299209110020
78. Casarett GW: A one-solution stain for spermatozoa. Stain Technol 1953;28:125-127. doi: 10.3109/10520295309105113
2. Peter AT, Brito LF, Althouse GC, et al: Andrology laboratory review: evaluation of sperm motility. Clinical Theriogenology 2021;13:297-315. doi: 10.58292/ct.v13.9359
3. Mortimer D, Menkveld R: Sperm morphology assessment: historical perspectives and current opinions. J Androl 2001;22:192-205. doi: 10.1002/j.1939-4640.2001.tb02171.x
4. Rodriguez-Martinez H: Semen evaluation and handling: emerging techniques and future development. In: Chenoweth PJ, Lorton SP: editors. Animal Andrology. Theories and Applications. Wallingford; CABI: 2014. p. 509-549.
5. Sathe S, Shipley CF: Applied andrology in sheep, goats and selected cervids. In: Chenoweth PJ, Lorton SP: editors. Animal Andrology. Theories and Applications. Wallingford; CABI: 2014. p. 226-253.
6. Mortimer D: The functional anatomy of the human spermatozoon; relating ultrastructure and function. Mol Hum Reprod 2018;24:567-592. doi: 10.1093/molehr/gay040
7. Encisco M, Cisale H, Johnston SD, et al: Major morphological sperm abnormalities in the bull are related to sperm DNA damage. Theriogenology 2011;76:23-32. doi: 10.1016/j.theriogenology.2010.12.034
8. Erenpreiss J, Spano M, Erenpreisa J, et al: Sperm chromatin structure and male fertility: biological and clinical aspects. Asian J Androl 2006;8:11-29. doi: 10.1111/j.1745-7262.2006.00112.x
9. Brito LF: Evaluation of stallion sperm morphology. Clin Tech Equine Pract 2007;6:249-264. doi: 10.1053/j.ctep.2007.09.004
10. 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. doi: 10.3109/10520290902984274
11. Chang V, Garcia A, Hitschfield N, et al: Gold-standard for computer-assisted morphological sperm analysis. Comput Biol Med 2017;83:43-150. doi: 10.1016/j.compbiomed.2017.03.004
12. Reeves A, Klabnik J, Strickland L, et al: Bull sperm morphology assessment varied by evaluator. Clinical Theriogenology 2022;14:11-17. doi: 10.58292/ct.v14.9293
13. Mortimer D: Practical Laboratory Andrology. Oxford; Oxford University Press: 1994. p. 393.
14. Chenoweth PJ: Sperm morphology. In: Chenoweth PJ, Lorton SP: editors. Manual of Animal Andrology. CABI: Wallingford; 2022. p. 45-55.
15. Hackett AJ, MacPherson JW: Some staining procedures for spermatozoa. A review. Can Vet J 1965;6:55-62.
16. Amann RP, Waberski D: Computer-assisted sperm analysis (CASA): capabilities and potential developments. Theriogenology 2014;81:5-17. doi: 10.1016/j.theriogenology.2013.09.004
17. Mortimer S, van der Horst G, Mortimer D: The future of computer-aided sperm analysis. Asian J Androl 2015;17:545-553. doi: 10.4103/1008-682X.154312
18. Keel BA, Stembridge TW, Pineda G, et al: Lack of standardization in performance of the semen analysis among laboratories in the United States. Fertil Steril 2002;783;603-608. doi: 10.1016/S0015-0282(02)03296-X
19. Moretti E, Sutera G, Collodel G: The importance of transmission electron microscopy analysis of spermatozoa: Diagnostic applications and basic research. Syst Biol Reprod Med 2016;62:171-183. doi: 10.3109/19396368.2016.1155242
20. Soley JT, du Plessis L: Ultra-imaging in applied animal andrology: the power and the beauty. Anim Reprod Sci 2020;220:106306. doi: 10.1016/j.anireprosci.2020.106306
21. Hancock JL, Trevan DJ: The acrosome and post-nuclear cap of bull spermatozoa. Royal J Microscop Soc 1957;76:77-83. doi: 10.1111/j.1365-2818.1956.tb00442.x
22. Kuster CE, Singer RS, Althouse GC: Determining sample size for the morphological assessment of sperm. Theriogenology 2004;61:691-703. doi: 10.1016/S0093-691X(03)00240-1
23. Mortimer D: A technical note on the assessment of human sperm vitality using eosin-nigrosin staining. Reprod Biomed Online 2020;40:851-855. doi: 10.1016/j.rbmo.2020.03.002
24. WHO laboratory manual for the examination and processing of human semen. 6th edition, WHO Press: 2021.
25. Kaya A, Birler S, Enwall L, et al: Determinants of sperm morphology. In: Chenoweth PJ, Lorton SP: editors. Manual of Animal Andrology. CABI; Wallingford: 2014. p. 34-56.
26. Blom E: Interpretation of spermatic cytology in bulls. Fertil Steril 1950;1:223-238. doi: 10.1016/S0015-0282(16)30183-2
27. Blom E: The ultrastructure of some characteristic sperm defects and a proposal for a new classification of the bull spermiogram. Atti del VII Simposia Int di Zootecnia. Milan 1972;7:125-139.
28. Rao AR: Changes in the morphology of sperm during their passage through the genital tract of bulls with normal and impaired spermatogenesis. PhD Thesis Royal Vet College, Stockholm: 1971.
29. Bjorndahl L, Mortimer D, Barratt CLR, et al: A Practical Guide to Basic Laboratory Andrology. Cambridge; Cambridge University Press: 2010. p. 336.
30. Saacke RG: Sperm morphology: its relevance to compensable and uncompensable traits in semen. Theriogenology 2008;70:473-478. doi: 10.1016/j.theriogenology.2008.04.012
31. Chemes HE, Rawe YV: Sperm pathology: a step beyond descriptive morphology. Origin, characterization and fertility potential of abnormal sperm phenotypes in infertile men. Hum Reprod Update 2003;9:405-428. doi: 10.1093/humupd/dmg034
32. Chenoweth PJ, McPherson FJ: Genetic aspects of male reproduction. In: Chenoweth PJ, Lorton SP: editors. Animal Andrology: Theories and Applications. CABI; Wallingford: 2014. p. 144-173.
33. Chenoweth PJ: Genetic sperm defects. Theriogenology 2005;64:457-468. doi: 10.1016/j.theriogenology.2005.05.005
34. Lorton SP: Evaluation of semen in the andrology laboratory. In: Chenoweth PJ, Lorton SP: editors. Animal Andrology. Theories and Applications. CABI; Wallingford: 2014. p. 100-143.
35. Cairo Consensus Workshop Group. The current status and future of andrology: a consensus report from the Cairo workshop group. Andrology 2020;8:26–52. doi: 10.1111/andr.12720
36. Brito LF, Greene LM, Kelleman, et al: Effect of method and clinician on stallion sperm morphology evaluation. Theriogenology 2011;76:745-750. doi: 10.1016/j.theriogenology.2011.04.007
37. Frenaeu GE, Chenoweth PJ, Ellis R, et al: Sperm morphology of beef bulls evaluated by two different methods. Anim Reprod Sci 2010;118:176-181. doi: 10.1016/j.anireprosci.2009.08.015
38. Menon AG, Thundathil JC, Wilde R, et al: Validating the assessment of bull sperm morphology by veterinary practitioners. Can Vet J 2011;52:407-408.
39. Chacón J: Assessment of sperm morphology in Zebu bulls, under field conditions in the tropics. Reprod Dom Anim 2001;36:91-107. doi: 10.1046/j.1439-0531.2001.00253.x
40. Cooper TG, Atkinson AD, Nieschlag E: Experience with external quality control in spermatology. Hum Reprod 1999;3:765-769. doi: 10.1093/humrep/14.3.765
41. Alvarez C, Castilla JA, Ramírez JP, et al: External quality control program for semen analysis: Spanish experience. J Assist Reprod Genet 2005;22:379-387. doi: 10.1007/s10815-005-7461-2
42. Wiltbank JN and Parish NR: Pregnancy rate in cows and heifers bred to bulls selected for semen quality. Theriogenology 1986;25:779-783. doi: 10.1016/0093-691X(86)90093-2
43. McPherson FJ, Nielsen SG, Chenoweth PJ: Semen effects on insemination outcomes in sows. Anim Reprod Sci 2014;151:28-33. doi: 10.1016/j.anireprosci.2014.09.021
44. Pukazhenthi BS, Wildt DE, Howard JG: The phenomenon and significance of teratospermia in felids. J Reprod Fertil Suppl 2001;57:423-433.
45. Waberski D, Riesenbeck A, Schulze M, et al: Application of preserved boar semen for artificial insemination: past, present and future challenges. Theriogenology 2019;137:2-7. doi: 10.1016/j.theriogenology.2019.05.030
46. Clark G: Staining Procedures. Biological Stain Commission. 4th edition, Baltimore; Williams and Wilkins: 1981.
47. Blom E: A one-minute live-dead stain by means of eosin nigrosin. Fertil Steril 1950;1:176-177. doi: 10.1016/S0015-0282(16)30125-X
48. Bjorndahl L, Soderlund I, Kvist U: Evaluation of the one-step eosin-nigrosin staining technique for human sperm vitality assessment. Hum Reprod 2003;18:813-816. doi: 10.1093/humrep/deg199
49. Saacke RG, Marshall CE: Observations on the acrosomal cap of fixed and unfixed bovine spermatozoa. J Reprod Fertil 1968;16:511-514. doi: 10.1530/jrf.0.0160511
50. Johnson L, Berndtsen WE, Pickett BW: An improved method for evaluating acrosomes of bovine spermatozoa. J Anim Sci 1976;42:951-954. doi: 10.2527/jas1976.424951x
51. Hanson R., Reddick S., Thuerauf S, et al: Comparison of bull sperm morphology evaluation methods under field conditions. Clinical Therigenology 2023;15:1-9. doi: 10.58292/CT.v15.9425
52. Morselli MG, Colombo M, Faustini M, et al: Morphological indices for canine spermatozoa based on the World Health Organization laboratory manual for human semen. Reprod Domest Anim 2019;54:949-955. doi: 10.1111/rda.13440
53. Zhu WJ: Preparation and observation methods can produce mispleading artefacts in human sperm ultrastructural morphology. Andrologia 2018;50:e13043. doi: 10.1111/and.13043
54. O’Connell M, McClure N, Lewis SEM: The effects of cryopreservation on sperm morphology, motility and mitochondrial function. Hum Reprod 2002;17:704-709. doi: 10.1093/humrep/17.3.704
55. Spindler R, Keeley T, Satake N: Applied andrology in endangered, exotic and wildlife species. In: Chenoweth PJ, Lorton SP: editors. Animal Andrology. Theories and Applications. CABI; Wallingford: 2014. p. 450-473.
56. Dey S, Kharbuli SM, Chakraborty R, et al: Effect of environmental acid-stress on the sperm of a hill-stream fish devario aequipinnatus: a scanning electron microscopic evaluation. Microsc Res Tech 2009;72:76-78. doi: 10.1002/jemt.20640
57. Erenpreiss J, Hlevicka S, Zalkalns J, et al: Effect of Leukocytospermia on Sperm DNA integrity: a negative effect in abnormal semen samples. J Androl 2002;23:717-723. doi: 10.1002/j.1939-4640.2002.tb02315.x
58. Althouse GC, Lu K: Bacteriospermia in extended porcine semen. Theriogenology 2005;63:573-584. doi: 10.1016/j.theriogenology.2004.09.031
59. Moretti E, Capitani S, Figura N, et al: The presence of bacteria species in semen and sperm quality. J Assist Reprod Genet 2009;26;47-56. doi: 10.1007/s10815-008-9283-5
60. Dowsett KF, Knott LM: The influence of age and breed on stallion semen. Theriogenology 1996;46:397-412. doi: 10.1016/0093-691X(96)00162-8
61. Rijsselaere T, Maes D, Hoflack G, et al: Effect of body weight, age and breeding history on canine sperm quality parameters measured by the Hamilton-Thorne analyser. Reprod Domest Anim 2007;42:143-148. doi: 10.1111/j.1439-0531.2006.00743.x
62. Hesser A, Darr C, Gonzales K, et al: Semen evaluation and fertility assessment in a purebred dog breeding facility. Theriogenology 2017;87:115-123. doi: 10.1016/j.theriogenology.2016.08.012
63. Johananison E, Campana A, Luthi R, et al: Evaluation of ‘round cells’ in semen analysis: a comparative study. Hum Reprod Update 2000;6:404-412. doi: 10.1093/humupd/6.4.404
64. Aitken RJ, Baker HWG: Seminal leukocytes: passengers, terrorists or good Samaritans. Hum Reprod 1995;10:1736-1739. doi: 10.1093/oxfordjournals.humrep.a136165
65. González-Marín G, Roy R, López-Fernández C, et al: Bacteria in bovine semen can increase sperm DNA fragmentation rates: a kinetic experimental approach. Anim Reprod Sci 2011;123:139-148. doi: 10.1016/j.anireprosci.2010.11.014
66. Keel BA, Webster BW: Handbook of the laboratory diagnosis and treatment of infertility. Boca Raton; CRC Press: 2000. p. 431.
67. Watson PF: Use of a Giemsa stain to detect changes in the acrosome of frozen ram semen. Vet Rec 1975;97:12-15. doi: 10.1136/vr.97.1.12
68. Williams WW, Savage A: Observations upon the seminal micropathology of bulls. Cornell Vet 1925;15:353-375.
69. Lagerlóf N: Changes in the spermatozoa and in the testes of bulls with impaired or enhanced fertility. Acta Pathol Microbiol Scand 1934;19: p. 254.
70. Oettle EE: Using a new acrosome stain to evaluate sperm morphology. Vet Med 1986;81:263-266.
71. Chan PJ, Corselli JU, Jacobson WC, et al: Spermac stain analysis of human sperm acrosomes. Fertil Steril 1999;72:124-128. doi: 10.1016/s0015-0282(99)00201-0
72. Sousa APM, Tavares RS, de la Calle JFV, et al: Dual use of Diff-Quik-like stains for the simultaneous evaluation of human sperm morphology and chromatin status. Hum Reprod 2009;24:28-36. doi: 10.1093/humrep/den365
73. Pozor MA, Zambrano GL, Runcin E, et al: Usefulness of dip quick stain in evaluating sperm morphology in stallions. Proc Am Assoc Equine Pract 2012;58:506-510.
74. Barth AD, Oko RJ: Abnormal Morphology of Bovine Spermatozoa. 1st edition, Ames, IA; Iowa State University Press: 1989. p. 302.
75. Erenpreiss J, Jepson K, Giwercman A, et al: Toluidine blue cytometry test for sperm DNA conformation: comparison with the flow cytometric sperm chromatin structure and TUNEL assays. Hum Reprod 2004;19:2277-2282. doi: 10.1093/humrep/deh417
76. Tsarev I, Bungum M, Giwercman A, et al: Evaluation of male fertility potential by Toludine Blue test for sperm chromatin structural assessment. Hum Reprod 2009;24:1569-1574. doi: 10.1093/humrep/dep068
77. Kovacs A, Foote RH: Viability and acrosome staining of bull, boar and rabbit spermatozoa. Biotech Histochem 1992;67:119-124. doi: 10.3109/10520299209110020
78. Casarett GW: A one-solution stain for spermatozoa. Stain Technol 1953;28:125-127. doi: 10.3109/10520295309105113
Published
2024-09-19
How to Cite
Chenoweth P., Brito L., Peter A., Waberski D., Althouse G., Aurich C., Luvoni G., Turner R., Fraser N., & Lopate C. (2024). Andrology laboratory review: evaluation of sperm morphology. Clinical Theriogenology, 16. https://doi.org/10.58292/CT.v16.10600
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Section
Review Reports
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