SPERM MOTILITY AND FERTILIZING ABILITY OF MEN WITH INFERTILITY I. Ilieva, S. Ivanova, P. Tzvetkova and Y. Gluhcheva Institute of Experimental Morphology and Anthropology with Museum,Bulgarian Academy of Sciences, Sofia, Bulgaria Correspondence to: Iliana Ilieva E-mail: iilieva@abv.bg ABSTRACT Motility is one of the main biological properties of spermatozoa that distinguishes them from the other cells in the organism. Their ability to move is of significant importance for the assessment of the ejaculate s quality - main criterion for determination of a man s fertility. The investigation of the factors effecting the motility of the spermatozoa aims to determine the relationship between the physico-chemical characteristics of the sperm, the pathology of the male reproductive tract and the mature spermatozoid. The results show that the highest percentage of spermatozoa with motility without progression is found in case of sexuallytransmitted infections (.25%), immotile spermatozoa are present mainly in case of specific inflammatory processes (44.5%) of the male reproductive system. The obtained data allow determination of spermatozoa motility rating as a marker for their fertilizing ability and for evaluation of the degree of infertility. In the present study we find that the percentage of spermatozoa with the highest mobility rating in cases of vascular and congenital diseases of the male reproductive tract is 55% and 51.6% respectively Keyword: fertility, infertility, motility rating, spermatozoa Introduction Male infertility is a result of various disorders of the reproductive system as well as of different biological factors regulating the reproductive process. Viscosity, ph, agglutination, presence of limited number of leukocytes or other white blood cells affect the progressive motility and fertilizing ability of the normal spermatozoa (). Morphology of mature spermatozoa is another main criterion that is directly related to their motility and fertilizing ability (1, 3, 5, ). Materials and methods 1221 ejaculates of infertile men (mean age 32.6 ± 3.5 years) with congenital, inflammatory and vascular diseases (Tabl. 1) of the male reproductive system were studied. The results were compared with those of 2 healthy male (mean age 3.12 ± 1.22 years) with preserved fertilizing capacity. TABLE 1 Distribution of the surveyed patients Patients with: Numb. Patients with: Numb. Congenital diseases of male sexual system 1 Nonspecific inflammatory diseases 39 Monorchism 15 Prostatitis chronica 25 Polyorchidism 1 Epididymitis chronica 15 Kryptorchism 14 Sexually transmitted infections - STI 55 Kysta epididymis 23 Vascular diseases 363 Inflammatory diseases of male sexual system 61 Varicocele 329 Specific inflammatory diseases 226 Torsio testis 34 Tuberculosis of genital tract TBC 69 Total number of patients 1221 Mumps orchitis MO 15 Control group healthy men 2 Spermatological analysis of the ejaculates, light- and transmission electron microscopy studies of the sperm cells 52
were performed according to the WHO criteria (16). For evaluation of the fertilizing capacity of men the index for progressive motility was used: М х N х V A x 1 were М number of mobile spermatozoa, expressed in percents; N number of spermatozoa, expressed in million per 1 ml ejaculate; V velocity of vertical movement, expressed in mm per 3 min; А percentage of pathological types of spermatozoa Results and Discussion Physico-chemical characteristics of the sperm and infertility Elevated ejaculate viscosity was found in cases of inflammatory diseases (2.2%) and in all cases of sexuallytransmitted infections (Fig. 1a). a b 1 9 6 5 4 3 2 1 9.1 2.99 CONGENIT AL 1.1 2.2 INFLAMMATORY 99.42 VASCULAR.5 Normal Elevated p H 9 6 5 4 3 2 1 A NORCHISM M ONORC HISM POLYORCH I D I SM K R YPTORC HIDIS M DEL-CASTILO SYNDROM K LI NEFELTER'S SY NDROM K Y S TA EPI DIDYMIS A GEN ES I A D. DEFERENS c d..6.4.2 6. 6.6 6.4 6.2..51 6.93 EPIDIDYMITIS chr. PROSTATITIS chr..19.62 ph 9 6 5 4 3 2 1.9.6.62 VARICOCELE TORSIO TESTIS контролна група Fig. 1 а) Viscosity and рн of ejaculate of patients with: b) congenital, с) inflammatory and d) vascular diseases of the reproductive system Our data as well as that of other authors show that seminal plasma viscosity is elevated in cases of inflammation and degenerative alterations of the testis (). It is determined that elevated viscosity is related not only to motility abnormalities of spermatozoa but to their concentration as well and is rarely found together with abnormal morphology (1). рн of ejaculate. The values (Fig.1b, 1c, 1d) of the alkaline-basic environment SPECIAL EDITION/ON-LINE 53 12 YEARS OF ACADEMIC EDUCATION IN BIOLOGY
of each ejaculate in case of pathology of the male reproductive system is in the range of.. and is comparable with the normal -.6. Agglutination of spermatozoa. Spontaneous agglutination of the spermatozoa was observed in 9.5% of the examined samples of male infertility. In cases of non-specific inflammatory diseases and STI mainly agglutination of mixed type was found (Fig. 2), and in individual cases head-to-head type was seen. The obtained percentage of agglutination from our data is in agreement with that of other authors (13, 14). Thakur et al. (13) observed this process in 12.5% and 1% of the cases of primary and secondary infertility, respectively, while Williams (14) showed spontaneous agglutination in about % of patients with decreased fertilizing capability. a b Fig. 2 а) Agglutination and b) Correlation between agglutination, motility and morphology of spermatozoa in ejaculates of patients with infertility. Results in Fig. 2b show proportional dependency between agglutination, motility and the percentage of spermatozoa with abnormal morphology in ejaculates of patients with increased microbial count of contamination in cases of inflammation. It is suggested that correlation between agglutination of spermatozoa and fertility is disputable (1, 11, 15). Other cell types in the seminal plasma Pyospermia (6, 12) is the main symptom in chronic infections of the reproductive system. In case of STI and prostatitis chr. (12.69%), epididymitis chr. (2.9%), kysta epididymis (5.56%) pyospermia is seen. In the rest of the examined groups 5- Leuc/field are counted (Tabl. 2). TABLE 2 Pyospermia and infertility Pathology to 1 Leuc/field over 1 Leuc/ field Pyospermia Viscosity Kysta epididymis (n=1) 5- Leuc/field - 1.12% 12 Leuc/ field -5.56% 5.56% Elevated - 22.23% TBC (n=69) 4-6 Leuc/ field -.24% - - - MO (n=114) 6- Leuc/ field -3.12% Single case - - Epididymitis chr. (n=15) - Leuc/ field -6.6% 15-16 Leuc/ field -.95% 2.6% Elevated 2.61% Prostatitis chr. (n=25) 4-6 Leuc/ field -2.49% 15-2 Leuc/ field - 11.12% 12.69% Elevated 4.3% STI (n=55) Single cases 1-15 Leuc/field 12.69% elevated - 1% Morphology of spermatozoa and infertility Different degenerative abnormalities in the spermatozoa 54
structure are observed in all disorders of the male reproductive system. The variety of specific morphological alterations is related to the immotility spermatozoa of infertile men. Our morphological studies showed that abnormalities in the head of spermatozoa (kryptorchidism 49%, varicocele 46%, torsio testis 49%) prevail to those of the tail (observed mainly in МО 31%) and to the mixed abnormalities (TBC 25%) (Tabl. 3). TABLE 3 Spermatozoa with abnormal configuration (%) in case of pathology of the male genital tract Abnormal gametes Kryptor chidism Kysta epididymis TBC MO Prostatitis chr. Epididymitis Varicocele Torsio testis chr. Head 49 29 1 21 39 42 46 49 Tail 21 31 9 19 2 2 Cytopl.droplet 2 2 1-3 2 - Mixeds 4 2 25 9 15 3 1 Total % 63 54 51 6 63 6 51 59 Motility, velocity and rating of spermatozoa Motility of spermatozoa is the main qualitative parameter for evaluation of their fertilizing potential. Despite pathology of the reproductive tract in congenital 33.5 %, inflammatory 13.33 % and vascular diseases - 2% we observed spermatozoa with progressive motility. In the rest of the cases decreased or no motility was registered (Tabl. 4). While evaluating the physico-chemical parameters of the ejaculate, morphology and motility of spermatozoa useful data for the motility rating of the sperm cells can be obtained for the different disorders of the male reproductive system (Fig. 3). TABLE 4 Motility/number and velocity of spermatozoa in case of disorders of the male reproductive system Pathology Motility (%) Velocity (µ/sec) Progressive Non-progressive motile Immotility Congenital diseases of male sexual system (n=11) Kysta Epididymis (n= 9) 33.5 ± 3.5 11.25 ± 1.24 55. ± 3.2 < 4 Kryptorchydism (n= 22) - 5 95-1 acinesis 3 Inflammatory diseases of male sexual system (61) Specific (n= 9) 13.33 ± 6.1 21.66 ±.3 65.1 ± 1.56 3. ±.9 Nonspecific 4. ± 1.12 1. ±.9 5. ±. 2.2 ± 2.22 Vascular diseases of male sexual system (34) Torsio testis (n= ) 2. ± 2.3 4. ± 1.65 4. ± 1. 6. ±.5 Control group healthy men (n=3) 63.9 ± 5.5 15.13 ± 1.6 2.9 ± 4.4 16.31 ± 1.3 The highest motility rating of spermatozoa was determined in case of vascular (55%), followed by that of congenital (51.6%) disorders of the genital tract. Conclusions and further perspectives The fertilizing capacity of spermatozoa depends not only on their total number or the number of cells with good motility, but also on the parameters characterizing their motility and morphology. At some point they become crucial for fertilization because along with the physico-chemical factors and the pathology of the reproductive system they form a SPECIAL EDITION/ON-LINE 55 12 YEARS OF ACADEMIC EDUCATION IN BIOLOGY
complex of spermatological markers for evaluation of the reproductive process of men. Fig. 3 Motility rating REFERENCES 1. Tzvetkova, P.D. 26. Ph. D. thesis. IEMAM-BAS, Sofia 2. Tzvetkova et al (2) Androl 1:14-21 3. Amann et al (1993) J Androl 14:39-46 4. Bellabara et al (2) Arch Androl 45:131-136 5. Hafez E (23) Male infertility. In: Humana Reprodukcija. Bojovic S (Ed) Beograd, pp 25-53 6. Hillier et al (199) Obstet Gynecol 5:-4. Hossain et al (199) Arch Androl 39:111-11. Klein et al (1991) Arch Path Lab 116:11-15 9. Kjaergaard et al (199) Acta Obstet Gynecol 6: 52-531 1. Mortimer et al (192) Arch Androl :165-11 11. Schirren, (191) Practical Andrology. Berlin: BRUDER Hartmann 12. Sigman et al (1993) J Urol1 49:133-134 13. Thakur et al (199) Ind J Pathol & Mcrobiol 41:343-35 14. Williams W (19) Fertil Steril 21:222-223 15. Wilson L (19) Proc Roy Soc Med 9:652-655 16. Word Health Organization: WHO Laboratory manual for the examination of Human semen and semen. New York: Cambridge University Press, 1992 56