Ronald L. Urry

A Comparison of Subjective Measurements of Human Sperm Motility and Viability with Two Live-Dead Staining Techniques*

Two commonly used live-dead stains (eosin-nigrosin (EN) and eosin-opal blue (EOB)) were compared with the estimated active spermatozoa in semen samples from patients attending an infertility service. Twenty-eight semen samples were analyzed throughout the day of their collection by estimating the number of active spermatozoa and by staining a portion of the incubated sample (37 degrees C) with each stain. The samples were analyzed 30, 60, 120, 240, and 360 minutes after the initial collection. At 30 minutes there were no significant differences between the estimated values and those of either stain. The slope of the EN stain closely paralleled that of the estimated measurements throughout the remainder of the time periods, while the EOB slope was somewhat steeper. Repeatability of values for semen samples obtained on different days was generally good for each stain. A comparison of semen samples from 85 patients, 30 minutes after collection, showed no significant differences between the numbers of estimated active sperm and the percentage live using the EN stain. Studies of 132 semen samples using the EN stain showed a positive correlation with the over-all quality of sperm motility. The results indicate that there is a role for live-dead staining in assessing semen quality.

Semen Analysis: A Review of Samples from 225 Men Seen at an Infertility Clinic

The results from 225 men attending an infertility clinic are presented. The percentage of oval, viable and active sperm cells, and the motility scores were lower in samples with counts less than 10 times 10(6) per ml., increased in counts or 10 to 40 times 10(6) per ml. and again increased in counts more than 40 times 10(6) per ml. The percentage of semen samples with abnormalities in measured parameters dramatically increased as the sperm count decreased. The percentage of samples with significant numbers of white blood cells was higher in samples with sperm counts less than 10 times 10(6) per ml. and in the azoospermic patients, while viscosity problems seemed to be associated with counts less than 10 times 10(6) per ml. Agglutination was more of a problem in samples with counts more than 40 times 10(6) per ml.

Inhibition of rat spermatogenesis and seminiferous tubule growth after short-term and long-term administration of a monoamine oxidase inhibitor.

Seminiferous tubules from rats killed 24 hours after injection of pargyline, a monoamine oxidase inhibitor, did not grow well in tissue culture when compared to control tubules. Treated tubules showed severe tubular degeneration and loss of cellular detail after nine days in culture. Animals injected with pargyline for ten days had varying degrees of semi-niferous tubule degeneration with depletion of the spermatogenic elements. It is suggested that pargyline has a detrimental effect on spermatogenesis. Pargyline possibly acts by decreasing MAO levels which, in turn, may increase potentially damaging amines which may be responsible for the testicular damage.

Correlation between follicle stimulating hormone luteinizing hormone testosterone and 5-hydroxyindole acetic acid with sperm cell concentration.

Plasma follicle stimulating hormone, luteininzing hormone, testosterone, urinary 5-hydroxyindole acetic acid and 17-ketosteroids were measured in patients seen at an infertility clinic. Plasma folicle stimulating hormone and luteinizing hormone levels, and urinary 5-hydroxyindole acetic acid levels were increased in patients with sperm concentrations less than 10 times 10(6) per ml. Plasma testosterone levels were lower in patients with sperm concentrations less than 10 times 10(6) per ml. The results suggest that in patients with sperm counts less than 10 times 10(6) per ml. there is not only impaired spermatogenesis but also decreased Leydig cell function. Urinary 17-ketosteroid levels were not related to sperm cell concentration.

Vasectomy and vasovasostomy. II. A comparison of two methods of vasovasostomy: silastic versus chromic stents.

Twelve adult, bilaterally vasectomized dogs were used to compare two different procedures for reuniting the severed vas deferens. Approximately 5 months postvasectomy six dogs had vasovasostomies utilizing Silastic stents and the vasa deferentia of six additional dogs were rejoined with intravasal chromic stents. Ejaculates were subsequently obtained at 2-week intervals and evaluated for sperm count, motility, viability, and seminal ion concentrations. All of the dogs in which Silastic had been utilized as the stent had sperm reappear in the ejaculate, whereas only 40% of the dogs in which chromic catgut had been utilized had reappearance of spermatozoa. The over-all quality of the ejaculate was also better in the animals in which Silastic had been utilized. Testicular and epididymal histology was normal in most dogs of both groups, but three of the dogs with chromic stents in which the anastomosis had not been a success had abnormal histology, with reduced numbers of spermatocytes, spermatids, and spermatozoa. The data suggest that Silastic stents are better than chromic stents for vas reanastomosis. They also suggest that some dog testes react negatively to vasectomy and do not recover by 1 year postvasectomy when vas patency is not reversed.

Treating the Subfertile Male Patient: Improvement in Semen Characteristics after Low Dose Androgen Therapy

Preliminary results suggest that the semen of selected patients can be improved after administration of low doses of fluoxymesterone and that the therapeutic regimen may be most beneficial for individuals with reduced sperm motility, abnormal semen morphology and/or low semen volume.

Simultaneous determination of human sperm morphology and viability: Simple office technique

Determinations of sperm morphology and viability are useful techniques for assessment of semen quality. In this report eosin-nigrosin is compared with hematoxylin-eosin for effectiveness in characterizing spermatozoa. The results indicate that eosin-nigrosin and hematoxylineosin give comparable results in differentiating spermatozoa. In addition, eosin-nigrosin allows the determination of the per cent of viable cells in each morphologic category. The eosin-nigrosin staining technique is a simple, rapid method which may give additional information in the assessment of male infertility.

Elevated Urinary Levels of 5-Hybroxyindole Acetic Acid and Its Relationship Among Levels of Plasma Follicle Stimulating Hormone, Testosterone and Testicular Pathology in Patients with Severe Oligospermia and/or Azoospermia

Plasma follicle stimulating hormone, testosterone and urine 5-hydroxyindole acetic acid levels were measured in 300 patients seen at an andrology clinic. Testicular biopsies were obtained from many of the severely oligospermic or azoospermic patients. Plasma follicle stimulating hormone concentrations were related inversely to sperm cell concentration, while plasma testosterone concentrations were depressed as the sperm count decreased to 5 times 10(6) per ml. The urine levels of 5-hydroxyindole acetic acid followed the same pattern as follicle stimulating hormone and were elevated in the group of individuals with sperm counts less than 10 times 10(6) per ml. Elevations in 5-hydroxyindole acetic acid often were associated with abnormalities in the testicular biopsy. Only 30 per cent of the patients who had elevated 5-hydroxyindole acetic acid also had elevated follicle stimulating hormone or depressed testosterone levels. The factors responsible for the elevation in 5-hydroxyindole acetic acid levels as the sperm count declines remain unclear.

Effect of amylase on sperm motility and viability.

The effect of increasing concentrations of amylase on the percentage of active spermatozoa, the quality of their motility and the percentage of viable cells was studied in semen samples in vitro. The amount of amylase needed to liquefy viscous semen samples in vitro also was determined. The percentage of active spermatozoa and viable cells, and the quality of sperm motility were altered in relationship to the amylase levels. Significant decreases in these parameters compared to control values were seen at the higher concentrations of amylase. The lowest level of amylase did not alter these parameters significantly and was sufficient to liquefy 80 per cent of the viscous semen samples. Amylase appears to be effective at low concentrations for use in liquefying viscous semen samples, thus making them easier to analyze during routine semen examination. The level of amylase used and the interval between addition and analysis must be controlled carefully.

Supravital staining of spermatozoa: relationship of eosin concentration to the percentage of cells staining live.

Supravital staining of human spermatozoa is a useful technique to assess semen quality. We compared 3 concentrations of eosin (1, 2.5 and 5 per cent) for their effectiveness to differentiate viable and non-viable spermatozoa. The percentage of viable cells determined by each concentration was compared as well as the percentage of cells estimated to be active. The results indicate that the percentage of spermatozoa determined to be viable with the supravital stains can be altered by changing the percentage of eosin in the stains. Use of 1 per cent eosin gave values that were significantly higher than the percentage of cells determined to be viable with 5 per cent eosin and the percentage of cells estimated to be active. Better quality slides were produced with 5 per cent eosin, which provided values that correlated favorably with motility estimations.