AZOOSPERMIA CYTOLOGICAL MANIFESTATIONS

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1 ý Comptes rendus de l Académie bulgare des Sciences ÌÓÑ ÆÓ ¾¼½½ BIOLOGIE Morphologie AZOOSPERMIA CYTOLOGICAL MANIFESTATIONS Stefka Ivanova, Petia Tzvetkova (Submitted by Corresponding Member J. Jordanov on February 2, 2011) Abstract Several clinical factors for azoospermia exist. Records of semen analyses frequently contain the item round cells without further specification of the type of cells. We have investigated 1333 patients (on an average ± year-old) with congenital (n = 299), specific (n = 226) and nonspecific inflammatory (n = 390), and vascular diseases (n = 363) of male genital system and 129 (on an average ± 2.15-year-old) healthy men as a control group. The following methods were used: andrological anamnesis and status; spermatological analysis of the ejaculate and sperm morphology according to WHO [ 1 ]; cytological analysis of round cells using Papanicolaou staining technique; statistical significance was verified with Student s t-test and SPSS computer programme. Azoospermia was proved in 20.18% in all male genital pathology. Cytological analysis of type round sperm cells determined the following round cells of spermatogenic origin: spermatids 79%, spermatocytes 7%. The round cells of non-spermatogenic origin were counted: monocyte 2%, granulocyte 1%, macrophage 3%, abnormal form cells 8%. Knowledge of the clinical features and the cytological sight of azoospermia open new horizons in the treatment of infertility in some forms of azoospermia, and therefore knowledge of the specific degree of the testicular damage is a necessary step in the evaluation of azoospermic men. Key words: azoospermia, frequency, round cells, men Introduction. Several aetiologies for azoospermia exist, but the prospects of fertility in every case are very poor [ 2 4 ]. Azoospermia is found in approximately 5 20% of men evaluated for infertility [ 5, 6 ]. Records of semen analyses frequently contain the item round cells [ 7 ] without further specification of the type of cells. The round cells observed in the 1291

2 semen samples could be either of spermatogenic origin [ 8 10 ] or varying types of cells of nonspermatogenic origin [ ]. The purpose of this report is to provide an update on the clinical manifestation and frequency of azoospermia, also to investigate the cytological manifestation of such seminal plasma. Material and methods. We have investigated 1333 patients (on an average 24.81±1.90-year-old) with congenital, inflammatory and vascular diseases of male genital system and 129 (25.63 ± 2.15-year-old) healthy men as a control group (Table 1). T a b l e 1 Patients with diseases of male genital system PATIENTS WITH: CONGENITAL DISEASES OF MALE REPRODUC- TIVE SYSTEM NUMBER 299 CONGENITAL DISEASES OF TESTIS 276 Monorchism 15 Kryptorchism 148 Del-Castilo syndrome 32 Klinefelter s syndrome 81 CONGENITAL DISEASES OF EPIDIDYMIS 23 Kysta epididymis 23 INFLAMMATORY DISEASES OF MALE REPRO- DUCTIVE SYSTEM 671 SPECIFIC INFLAMMATORY DISEASES 226 Tuberculosis of genito-urinarae tract 69 Mumps orchitis 157 NONSPECIFIC INFLAMMATORY DISEASES 390 Epididymitis chronica 105 Prostatitis chronica 285 SEXUAL-TRANSMITTED INFECTION (STI) 55 VASCULAR DISEASES OF MALE REPRODUCTIVE SYSTEM 363 Varicocele 329 Torsio testis 34 CONTROL GROUP 129 We used the following methods: Andrological anamnesis and status, spermatological analysis of the ejaculate and sperm morphology according to [ 1 ]; 1292 S. Ivanova, P. Tzvetkova

3 Fig. 1. Spermatogenic cells in the seminal plasma on azoospermic men. (a e) spermatid cells; (f h) spermatocytes. Papanicolau, 600 Cytological analysis of seminal plasma, used the Papanicolaou staining to distinguish round cells of spermatogenic and nonspermatogenic origin. The following forms were identified: spermatogenic spermatogonia (dark and pale), primary spermatocytes, secondary spermatocytes, early and late spermatids (corresponding to Sa-Sb and Sc-Sd steps of spermatogenesis); nonspermatogenic round cells. Results. Cytological examination of seminal plasma in cases with azoospermia were studied. We observed cells divided into two groups (Fig. 1) spermatogenic and nonspermatogenic round cells (Fig. 2). Spermatids are the most common cell line spermatogenic (79%). As for those of nonspermatogenic origin, most common are monocytes (2%) and macrophages (3%) (Fig. 3). Discussion. Clinical manifestation and frequency of azoospermia 20.18%, described by us are not different according to other authors [ 14 16, 3, 4 ]. Inflammatory and vascular diseases of male reproductive system were very often genital pathology attended with disturbances of sperm fertilizing ability. Azoospermia was found in 20.35% on specific, 5.38% in nonspecific inflammatory Compt. rend. Acad. bulg. Sci., 64, No 9,

4 Fig. 2. Nonspermatogenic cells in the seminal plasma on azoospermic men. (a) degenerating monocyte, (b) monocyte, (c d) epithelial cells, (e) lymphocyte, (f) macrophage, (h) seminal plasma (azoospermia), (g) seminal plasma (normal, 400). Papanicolau, 600 diseases. Sexually-transmmited inflammation of male system led to 1.23% absence of spermatozoa. We proved varicocele and azoospermia in 19.72%. In our study we conducted cytological examination of seminal plasma in cases with azoospermia. Spermatids were the most frequently counted 79% of all types of cells. A number of spermatocytes in the semen is seldom accompanied by a large number of mature spermatozoa and severe disturbances of the spermatogenesis can be expected in such cases. On the other hand, degenerated spermatids are frequently observed in the presence of mature spermatozoa in the semen [ 17, 10 ]. According to Holstein and Schirren [ 9 ] abnormal form cells were detected in 50% of all cases studied. We found 8%. Atanassova and Kancheva [ 18 ] reported that chemical agents (vitamin A- deficiency), heat, and certain pathological conditions also affect selectively more differentiated germ cell stages. A differentiation of the round cells into cells of spermatogenic and nonspermatogenic origin is very important for a correct semen analysis. The lumping of 1294 S. Ivanova, P. Tzvetkova

5 Fig. 3. Frequency of nonspermatogenic cells in the seminal plasma on azoospermic men all round cells into one group (as it is suggested in many recommendations for semen analysis) highly increases the risk of misinforming the treating clinicians. Conclusion. Knowledge of the clinical features and the cytological sight of azoospermia open new horizons in the treatment of infertility in some forms of azoospermia, and therefore knowledge of the specific degree of the testicular damage is a necessary step in the evaluation of azoospermic men. REFERENCES [ 1 ] World Health Organisation (WHO). Special programme on research, development and research training in human reproduction. Annual Tech. Report., Geneva, WHO, [ 2 ] Matsumiya K., M. Namiki, S. Takahara. International Journal of Andrology, 17, 1994, No 3, [ 3 ] Popken G., J. Schwarzer. Urologie, 47, 2008, No 12, [ 4 ] Ferhi K., R. Avakian, J. Griveau, F. Guille. Andrologia, 41, 2009, No 2, [ 5 ] Jarow J., R. Oates, J. Buch, S. Shaban, M. Sigman. Urology, 49, 1992, [ 6 ] Foresta C., M. Zorzi, C. Galeazzi, M. Rossato. International Journal of Andrology, 18, 1995, [ 7 ] Johanisson E., A. Campana, R. Luthi, A. Agostini. Human Reproduction, 6, 2000, No 4, [ 8 ] Fawcett D., C. Marhert, J. Papanconstantinou. The Developmental Biology of Reproduction. New York, Academic Press, USA, 1975, Compt. rend. Acad. bulg. Sci., 64, No 9,

6 [ 9 ] Holstein A., C. Schirren, D. Fawcett, J. Bedford. The Spermatozoon: Maturation, Motility, Surfaces, Properties and Comparative Aspects. Baltimore, USA, Urban and Schwarzenberg, 1979, [ 10 ] Comhaire F., L. Vermeulen. Hum. Reprod., Update, 4, 1995, No 1, [ 11 ] Fedder J. Arch. Androl., 36, 1996, No 1, [ 12 ] Thomas J., S. Fishel. J. Hall. Hum. Reprod., 12, 1997, No 1, [ 13 ] Sapundjiev E. Practicum of Cytology and Histology. Sofia, GERA-ART, [ 14 ] Johannisson E., R. Eliasson. Int. J. Androl., 7, 1978, [ 15 ] Sousa M., A. Barros, K. Takahaski. Hum. Reprod., 14, 1999, No 5, [ 16 ] Uchechukwu I., O. Ezeh. Human Reproduction, 15, 2000, No 11, [ 17 ] Dym M., D. Fawcett. Biol. Reprod., 4, 1971, [ 18 ] Atanassova N., L. Kancheva. Compt. rend. Acad. bulg. Sci., 49, 1996, No 6, Department of Experimental Morphology Institute of Experimental Morphology, Pathology and Anthropology with Museum Bulgarian Academy of Sciences Acad. G. Bonchev Str., Bl Sofia, Bulgaria stefi13831@ .bg 1296 S. Ivanova, P. Tzvetkova

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