INTERFERENCE OF HUMAN SPERMATOZOA MOTILITY BY ESCHERICHIA COLI*

Transcription

1 FERTnITY AND STERILITY Copyright ~ 1971 by The Williams & Wilkins Co Vol 22 No5 May 1971 Printed in USA INTERFERENCE OF HUMAN SPERMATOZOA MOTILITY BY ESCHERICHIA COLI* NELSON S TEAGUE MDt SAUL BOYARSKY MD FACS* AND JAMES F GLENN MD FACS Division of Urology Department of Surgery Duke University Medical Center Durham North Carolina The association of pathogenic bacteria in the male genitourinary tract with lowered male fertility I has been noted in urologic and fertility-sterility textbooks but there is a scarcity of information as to why this is so Little is known regarding the effect of bacteria on infertility or on spermatozoa Since the early 1930's there have been sporadic references in the literature to the agglutination of spermatozoa by various bacteria and their endotoxins In 1962 Dennis 2 demonstrated immobilization of ram and bull spermatozoa by the endotoxin obtained from Vibrio fetus a common pathogen in rams and bulls Based on their interesting observations in animals and our observation that Escherichia coli and its endotoxin alters ureteral function 3 we devised a series of simple experiments to determine what effect a common human urinary tract pathogen might have on human spermatozoa METHOD Experiments were initiated no later than 1 hr after collection on ejaculate specimens obtained by masturbation from healthy adult volunteers 05-cc volumes of fresh ejaculate were mixed and diluted with 05-cc volumes of each of the following: (1) normal saline (2) sterile culture * Presented at the 26th Annual Meeting of the American Fertility Society Washington DC March t Present address: Jefferson Surgical Clinic Roanoke Va * Present address: Professor and Chairman Genito-Urinary Surgery Washington University School of Medicine St Louis Mo 281 broth media (3) live E coli suspension and (4) saline solutions containing E coli endotoxin (Difco 0127-B8) Untreated ejaculate served as a control for the other preparations Following mixing a drop of each of the suspensions was placed on a glass slide and examined under the microscope to determine motility and viability These examinations were repeated at hourly intervals for 8 hr with a final observation being made at hr The numerical system devised by Emmens 4 was employed for the grading of sperm motility and is outlined in Table 1 The E coli were obtained from the infected urine of patients Very heavy growths of the bacteria were utilized usually in excess of organisms/ml A total of 46 tests were conducted and separated into the following groups (Table 2) RESULTS From the onset it was apparent that the live bacteria markedly reduced the motility and viability of the spermatozoa (Fig 1) The curve representing the spermatozoa mixed with the live bacteria falls far below the other curves Immediately upon mixing there was a significant dampening of the activity the motility rating falling between 20 and 25 At 20 hr there was a complete absence of motility and viability in this group The promptness with which the decreased motility was observed to occur in the live E coli group was striking Immediately after mixing the ejaculate with the

2 282 TEAGUE ET AL Vol 22 Grade TABLE 1 Index of Motility Characteristics 40 Full activity but there may be up to 30-40~o of dead cells 35 A detectable dampening of activity compared with Sluggish rate of motion about 2!3 that of Most cells progressing but many stationary with tails vibrating 20 Most cells stationary many with tails vibrating 1 5 Many motionless none or almost none actively progressing 10 Hardly any motility only tails moving (very rarely or occasional) O 5 Only a few cells per field showing any movement Completely motionless o 0 TABLE 2 Group Preparation No of tests 1 Ejaculate alone 8 2 Ejaculate + broth 10 3 Ejaculate + saline 9 4 Ejaculate + endotoxin 8 5 Ejaculate + live bacteria 11 live bacteria clumping of the spermatozoa was noted (Fig 2) Interestingly no agglutination was noted in two experiments in which the bacteria were killed by boiling for 10 min and mixed with the ejaculate Findings similar to ours have previously been reported by Matthews and Buxton 5 who found that certain bacteria cultured from the cervix were spermicidal To verify this we obtained E coli bacteria from cervical cultures and repeated our experiments utilizing this organism The results were identical (Fig 3) Again the dramatic and immediate clumping of the spermatozoa was observed (Fig 4) DISCUSSION Certainly it seems evident that E coli which are pathogenic for humans can significantly depress the motility and viability of human spermatozoa in vitro The mechanism by which this alteration takes place is not obvious Many organic substances have been observed to influence mammalian spermatozoa Follicle cyst fluid tubal fluid and peritoneal fluids have the ability to accelerate the movement of human spermatozoa 6 Similarly prostatic secretions in the dog activate markedly the motility of spermatozoa 7 Alterations in acidity especially lowering ph are known to decrease the viability of spermatozoa ph determinations were obtained in the above experiments and in only one instance was ph noted to be less than 70 The normal ph of human semen is Therefore it would appear that ph alterations do not contribute to the depressed motility and decreased viability we have observed Our observations suggest that the decreased motility is a result of the rapid :S '" -- II: " ~ 20 ' -' ' e _ A Ejaculate + Endotoain r )( Ejaculat + Broth a Ejaculof + SaUne -'---t--- -Ejaculo +EColi -JC- - )C--X- Ejocu te alone 05 I! ' - 0L-'--2r-T3-'4r-T5--6r-T7-'8r-T9~~~~20 40 Full activit)' 35 A detectable dampinq of acllvlt 30 Slutllth motion 25- Man Itatlono cliltail' 'libra tin Time in Hours 20 Mot cli Itatiano) tall 'libra tin 15 Mon motlonl 10 Onl taill moving 015 F tail mowlnt onl 00 - Complt motion I FIG 1 Curve representing ejaculate plus urinary tract E coli demonstrates depressed motility For further information see Table 1 and Reference 4 (From Emmens C W J Physiol (London) 106: )

3 May 1971 SPERMATOZOA MOTILITY 283 FIG 2 Photograph on the left represents spermatozoa plus saline Photograph on right represents spermatozoa plus urinary tract E coli Clumping of spermatozoa prevents progressive motility clumping or agglutination of the sperma- saccharide of E coli failed to produce sigtoza Similar sperm agglutination phe- nificant alterations was surprising Hownomena have been observed when sper- ever this does not mean endotoxins are matozoa are mixed with certain viruses 8 40 and mycoplasmas 9 It is postulated a reejacula' + Saline ceptor mechanism is involved in this sperm 35 f adsorption to the mycoplasmas Perhaps antigen antibody-like reactions are in30 volved resulting in the clumping of the spermatozoa Also spermatozoa have been noted to be massively adsorbed to cells in~ 20 ::::; fected with viruses is 15 Normal human semen clots immediately following ejaculation but liquifies within min due to a proteolytic enzyme derived from the prostate gland One can 05 only speculate at this point that the live 1L---~~r-~3--~4--~--~6~~ E coli may interfere in some manner with TIME IN HOURS the semen enzymes or cellular membrane systems FIG 3 Curve representing ejaculate plus cervical The fact that the endotoxin or lipopoly- E coli demonstrates depressed motility

4 284 TEAGUE ET AL Vol 22 without effect on human spermatozoa since domonas Proteus and Gonococcus endotoxin from only one strain of E coli Therefore it would be worthwhile to was employed Also in our hands the re- search for genital infections especially actions in other biologic systems to endo- E coli in patients with unexplained intoxins varies tremendously from day to day fertility Meares and StameylO have shown with their split-urine cultures that bacterial insummary fections of the prostate do occur and not Live pathogenic E coli obtained from infrequently We would expect to find inurinary or cervical cultures produce profertility problems associated with genital found depression in the motility and VIinfections since we have shown the proability of human spermatozoa in vitro found influence live E coli have on spermatozoa Quesada Dukes Deem and REFERENCES Franklin l l reported on 50 patients who L LANE RoBERTS C Sterility and Impaired Fertility had fertility problems associated with (ed 1) Harper & Row New York 1948 p 78 genital infections They noted an improve- 2 DENNIS S M Spermicidal activity of bacterial ment of semen quality following eradicaendotoxin Nature (London) 195: tion of infection Schirren and Fander 12 3 TEAGUE N S AND BOYARSKY S Further effects of coliform bacteria on ureteral peristalsis J found decreased sperm motility in paurol 99: tients with proven E coli infections but 4 EMMENS C W The motility and viability of rab not in those with other organisms such bit spermatozoa at different hydrogenion conas Staphylococcus Streptococcus Pseucentration J Physiol (London) 106: ~ " " ' FIG 4 Photograph on the left represents spermatozoa plus saline Photograph on right represents spero matozoa plus cervical E coli Clumping of spermatozoa prevents progressive motility

5 May 1971 SPERMATOZOA MOTILITY 285 ' ' 5 MATTHEWS C S AND BUXTON C L Bacteriology of the cervix in cases of infertility Fertil Steril 2: BALIN H Follicular and tubal fluids in the reproductive process Amer J Obstet Gynec 76: IVANOV1 1 AND KASSAVINA B S Action of prostatic secretion on the motility and metabolism of spermatozoa Nature (London) 158: PELEG B A AND lanconescu M Spermagglutination and spermadsorption due to mycoviruses Nature (London) 211 : TAYLOR-RoBINSON D Spermadsorption and spermagglutination by mycoplasmas Nature (London) 215: MEARES E M AND STAMEY T A Bacteriologic localization patterns in bacterial prostatitis and urethritis Invest UroI5: QUESADA E M DUKES C D DEEM G H AND FRANKLIN R R Genital infections and sperm agglutinating antibodies in infertile men JUral 99: SCHIRREN C AND FANDER H A Genitalinfektionen des Mannes und ikre Ausiverkungen aut die Spermalozoenmotilitat Med Welt 1: "