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Factors affecting sperm motility. I. In vitro change in motility with time after ejaculation.

Semen specimens from 33 donors were analyzed intermittently between 1 and 24 hours after ejaculation. Motility was determined objectively by analyzing still-camera photomicrographs taken with the aid of the new multiple exposure photography (MEP) technique, from which the following information was obtained: the percentage of motile spermatozoa, the average velocity and frequency distribution of spermatozoal velocities, and index of motility as well as sperm concentration. From curves drawn for each of the 33 analyzed specimens, these conclusions were drawn: 1. The percentage of motile spermatozoa decreases progressively, beginning 1 hour after ejaculation, at a rate of about 5% to 10%/hour. 2. In the majority of cases, sperm velocity increases for the first 4 hours and then decreases gradually. 3. There is no unique pattern of sperm motility and velocity with time for all specimens, especially during the first 4 hours. However, three typical curves of motility change were found during the first 4 hours: increase in motility, moderate decline in motility, and rapid loss of motility. 4. The frequency distribution of velocities of motile spermatozoa revealed typical normal curves at all times, but these curves became flatter as time elapsed. An explanation for these divergent patterns and the possible role of these findings in some physiologic and clinical problems are discussed.

Ejaculation↗

Factors affecting sperm motility. II. Human sperm velocity and percentage of motility as influenced by semen dilution.

Spermatozoal velocity and percentage of motility were analyzed objectively with the multiple exposure photography method before and after specimens from fertile and infertile men were diluted in their own seminal plasma or normal saline. No significant change in percentage of motility was found in samples diluted up to 1:6 in both kinds of diluents. However, a significant relative increase (up to 25% of the original velocity) was found when a specimen was diluted with its own seminal plasma, and an even greater increase (up to 37% of the original velocity) was found when it was diluted with saline. Compared with undiluted specimens, there was no delayed effect on spermatozoal motility when semen was diluted with saline after up to 4 hours' incubation time. Contrary to the findings in animal and human semen described by others, there was no deleterious effect on sperm motility with this kind and rate of dilution and duration of time. The assumption that the increase in sperm velocity caused by dilution is not excitatory but is due only to a decrease of seminal fluid viscosity and a reduced number of spermatozoa which interfere with sperm free movement is discussed. We recommend evaluation of spermatozoal motility in diluted specimens in addition to evaluation of the original specimen in any routine semen analysis in order to determine true spermatozoal motility potential under optimal conditions.

Culture Media↗

Factors affecting sperm motility. III. Influence of visible light and other electromagnetic radiations on human sperm velocity and survival.

Specimens of semen from fertile and infertile patients were exposed to different electromagnetic radiations, including visible light, ultraviolet (UV) light, x-rays, and high-frequency radio waves. Sperm motility was analyzed before, during, and after irradiation by the multiple exposure photography (MEP) method. No significant difference was found between controls and specimens exposed to various doses of visible and UV light and x-rays either immediately or several hours after exposure. In contrast to spermatozoa of other species that were reported to be adversely affected by visible and UV light and x-rays, human spermatozoa seem to be highly resistant to similar doses of these radiations. A deleterious influence was observed when high-frequency radio waves were applied to human spermatozoa. This may be attributed to an intracellular diathermic effect. The informative value of this study in relation to routine semen analyses and experimental studies in the physiology and comparative anatomy of spermatozoa is discussed.

Cell Survival↗

Factors affecting sperm motility. VI. Sperm viability under the influence of bacterial growth in human ejaculates.

The influence of bacterial growth on human sperm motility and viability was evaluated objectively with the multiple-exposure photography method. Experimental semen specimens, obtained from normal donors bh nonaseptic means of masturbation, were incubated with antibiotics at room temperature or body temperature for 24 hours. Although bacteria, grew in control specimens, were totally eradicated in all antibiotic-treated specimens, no significant difference was found between these groups with regard to sperm motility throughout the time of incubation. Sperm survival was not inhibited, nor was it extended as a result of suppression of bacterial growth. In both groups, survival time was much shorter in specimens incubated at body temperature than in those kept at room temperature. Sperm motility was not affected after 2 hours of incubation of fresh specimens with concentrations of various pathogenic bacteria similar to those found in severe prostatitis. The question of whether the use of antibiotics in the treatment of asthenospermia per se has a prognostic value is discussed.

Adult↗

Factors affecting sperm motility. VII. Sperm viability as affected by change of pH and osmolarity of semen and urine specimens.

The effects of pH and osmolarity of semen and urine specimens on motility and velocity of human spermatozoa were studied objectively with the aid of the multiple exposure photography (MEP) method. The pH of fresh ejaculates ranged from 7.2 to 8.2 and specimens were slightly hyperosmotic ranging between 300 to 380 mOsm/kg. Gradually changing the pH and osmolarity to either side of normal values led to progressive loss of sperm motility. However, sperm velocity was slightly increased by mild alkalinization and hyperosmolarity. Spermatozoa that became immobilized by acidification regained their motility shortly after pH was restored to normal values. In the majority of instances spermatozoa lost their motility when mixed with fresh urine specimens. Neutralization of urinary pH could not protect them from this effect unless urine osmolarity was also isotonically adjusted. It is suggested that patients with retrograde ejaculation should adequately increase their fluid intake before recovery of sperm from their bladder for artificial insemination.

Humans↗

Factors affecting sperm motility. IX. survival of spermatozoa in various biological media and under different gaseous compositions.

The role of various environmental conditions on sperm motility and their survival was investigated by incubating washed human spermatozoa in various biologic or artificial media and under different atmospheric compositions. When sperm were analyzed by the multiple exposure photography technique for objective motility determination, it was found that none of the investigated biologic media induced any immediate stimulatory effect on the tested spermatozoa. Exchange of seminal fluid between normal and oligoasthenospermic specimens did not reveal any beneficial or harmful effect on sperm motility. Sperm did not survive for more than 24 hours when kept in their original seminal fluid under any circumstances. The most favorable media tested for prolonged sperm survival were pure human umbilical cord serum or Ham's solution containing 0.5% human albumin. However, even in these media sperm did not survive much beyond 24 hours unless incubated in an atmosphere containing 5% CO2 or a triple mixture of 90% N2, 5% O2, and 5% CO2. Only under these circumstances could sperm stay alive, in some cases up to 5 days.

Carbon Dioxide↗

Computerized staged-freezing technique improves sperm survival and preserves penetration of zona-free hamster ova.

Sperm cryosurvival was evaluated after a staged freezing program, compared with standard vapor-cooling. Fifty-nine ejaculates produced by nine proven fertile males were studied before and after preservation with glycerol or dimethylsulfoxide. Spermatozoal survival was assessed by after-thaw motility, mean sperm velocity (MSV) with the use of multiple exposure photography and by the ability to penetrate zona-free hamster ova. A significantly greater reduction in sperm motility was observed with the addition of DMSO in contrast to G (P less than 0.01). All freezing methods resulted in reduced sperm motility, compared with unfrozen samples. The after-thaw sperm motility was significantly lowered when vapor-freezing was used in comparison with the staged process (22.9% +/- 2.6% versus 32.9% +/- 3.4%; P less than 0.01). The MSV was slightly, but not significantly, lower in the after-thaw semen, compared with fresh ejaculates. A significant impairment in sperm penetration ability was evidenced with vapor-freezing, compared with the staged cooling (25.7% +/- 3.1% versus 34.9% +/- 3.0%; P less than 0.01). The programmed freezing process rendered the same sperm efficiency in penetrating zona-free hamster ova as did the fresh samples. This relatively rapid, simple, and reliable computerized cooling technique improves the efficiency of sperm cryostorage and preserves its penetration capacity.

Animals↗

Enhancement of human sperm motility and velocity in vitro: effects of calcium and creatine phosphate.

Because of their roles in motility regulation and energy transport, calcium and creatine phosphate were examined for their effects on sperm motility and velocity in specimens of normal donors. Semen or migrated sperm fractions were incubated with of 1 mmol of calcium, 5 mmol magnesium, and 10 mmol of creatine phosphate (n = 28) or in the presence of 4 mumol of Verapamil, calcium, and creatine phosphate (n = 10). The samples were subjected to multiple exposure photography (four picture frames of two different drops) at 0, 1, 4, or 5 and at 10 hours and sperm motility and velocity were analyzed. In both calcium and calcium-creatine phosphate conditions, sperm motility and velocity were significantly increased, compared with control values (P = between less than 0.001 and 0.05). Sperm motility declined following Verapamil exposure, but the motility values remained at the level of the control in the presence of additional calcium or creatine phosphate. The effects of calcium and creatine phosphate take place rapidly; within 1 minute all improvements in sperm velocity and motility are fully achieved. There is no loading effect of calcium, and when the sperm is transferred into media without the additional calcium, the velocity decreased to that of the initial control value. Magnesium alone had no effect on motility or velocity. These experiments indicate that calcium or creatine phosphate can support sperm motility and velocity at a significantly increased level. Thus the addition of calcium or creatine phosphate to the insemination media may enhance the fertilizing capacity of sperm during in vitro fertilization or gamete intrafallopian transfer procedures.

Calcium↗

Follicular fluid enhances sperm motility and velocity in vitro.

Forty-three follicular fluids (FFs) obtained during laparoscopy were tested in vitro for their effect(s) on sperm motility using gametes obtained by the swim-up procedure. Both the proportion of motile sperm and the velocity distribution patterns were evaluated as function of time by multiple-exposure photography technique. At the various incubation periods considered, all FFs maintained or then enhanced sperm motility as compared with the paired control suspension incubated with a sperm survival medium. The results of the sperm contact test for FFs from women who achieved pregnancy versus FFs from women who remained infertile were not significantly different for both parameters measured. Comparing these with our previously reported results, we may hypothesize that FF released at ovulation into the peritoneal cavity may counteract some sperm-immobilizing effect of peritoneal fluid, thereby increasing the fertility potential of the male gametes.

Ascitic Fluid↗

Effect of volumetric mixtures of peritoneal and follicular fluid from the same woman on sperm motility and acrosomal reactivity in vitro.

OBJECTIVE: To study the effect of peritoneal (PF) and follicular fluids (FF from the same woman) as well as of given volumetric combinations thereof on sperm motility and acrosomal reactivity. DESIGN: Prospective. Peritoneal fluid and FF were incubated separately or in given volumetric combinations (PF/FF = 100/0, 75/25, 50/50, 25/75, 0/100; vol/vol) with swim-up sperm suspensions. SETTING: In vitro fertilization and general infertility clinic and laboratories. PATIENTS, PARTICIPANTS: Women participants of the gamete intrafallopian transfer program (motility study, n = 20; acrosomal reaction study, n = 14). Sperm donors of the artificial insemination program and men with given sperm parameters. INTERVENTIONS: Hormonal stimulation. Laparoscopy. MAIN OUTCOME MEASURES: Progressive velocity and percentage of motile gametes measured with multiple-exposure photography. Acrosomal reactivity measured by an immunofluorescent technique. RESULTS: Follicular fluid always influenced progressive motility and also sustained the number of motile gametes, as function of time, better than PF or the PF/FF mixtures (P less than 0.05). The acrosomal reactivity of sperm incubated in the various PF/FF combinations was low; after 5 hours only the FF-sperm suspensions showed a significant enhancement of acrosomally reacted gametes. CONCLUSION: At ovulation, FF transmit positive (motility- and acrosomal reactivity-enhancing) signals to sperm, whereas PF may transmit positive, neutral, or negative signals (noise signals). The volumetric combination of FF and PF in the tubal environment, which may differ from cycle to cycle and from woman to woman, could therefore result in synergic (or antagonistic) effects on the sperm fertility potential.

Acrosome↗

A new model for investigating in real-time the existence of chemotaxis in human spermatozoa.

OBJECTIVE: To suggest a new approach to the research of chemotaxis between various media and human spermatozoa to solve the riddle of its existence. DESIGN: Laboratory experiments in which chemotaxis between human spermatozoa and follicular fluid (FF) as well as N-formyl was investigated. SETTING: Male physiology laboratory and in vitro fertilization program unit. PATIENTS: Follicular fluid was collected from 15 patients and used either fresh or after storage at -5 degrees C for less than 2 weeks. High-quality semen specimens were obtained from normal donors and underwent one-step washing with Ham's F-10 solution (Biological Industries, Kibbutz Beth Haemek, Israel). TECHNIQUES: Assays were performed in real time by direct microscopical observation and quantitative determination of the disturbance in random movements of human spermatozoa caused by the tested media. Spermatozoa and tested media, including FF and N-formyl-Met-Leu-Phe were placed in a sealed minichamber and photographed with the aid of the multiple exposure photography technique. RESULTS: By analyzing photographed tracks of more than 80,000 moving spermatozoa, a physiologically insignificant deviation from randomness, corresponding to a 95% confidence interval of -0.57 +/- 0.46%, was found when spermatozoa swam along the concentration gradient of the tested media. The average velocity of spermatozoa swimming toward the test media was slightly lower (less than 5%) than that for spermatozoa swimming away, indicating, if anything, negative chemoattraction. CONCLUSIONS: Despite the high sensitivity of the model, neither natural FF nor synthetic N-formyl exerted any chemotaxis on human sperm. The advantages and potential use of this model for investigating chemotaxis by other biological and synthetic media are discussed.

Body Fluids↗

Use of a sealed minichamber for direct observation and evaluation of the in vitro effect of cigarette smoke on sperm motility.

OBJECTIVE: To determine the in vitro effect of cigarette smoke on sperm motility and survival in an attempt to find a possible association with clinical studies that had pointed to the existence of such an effect in vivo. DESIGN: Laboratory experiments in which the effect of cigarette smoke on human sperm placed in a sealed minichamber was directly observed and determined by motion analysis. PATIENTS: High-quality fresh semen specimens were obtained from healthy donors and used for the experimental studies. INTERVENTIONS: Various doses of filtered and nonfiltered cigarette smoke, collected by an improvised manual smoking machine, were streamed at a rate of 2 mL/s into sealed chambers containing samples of washed spermatozoa. Evaluation of the changes in sperm motility was performed in real time by direct microscopical observation and repeated determinations by the multiple exposure photography technique until their complete immobilization. RESULTS: Streaming of 100 mL of cigarette smoke induced a dramatic drop in sperm motility, which caused sperm immobilization in about 15 minutes. This effect showed a clear dose-response relationship either with the amounts streamed or with the time during which sperm were exposed to cigarette smoke. The harmful effect was almost the same whether sperm were exposed to filtered or nonfiltered cigarette smoke. The role of carbon monoxide and hydrogen cyanide on these findings was studied separately, and it was shown that the inhibitory effect was not due to the presence of these two important components of cigarette smoke. CONCLUSIONS: The clear evidence of the in vitro harmful effect of crude cigarette smoke on sperm motility points to the possibility that in heavy smokers certain toxic agents may reach mature spermatozoa or their precursors via the blood stream and induce their harmful effect on sperm motility in vivo.

Carbon Monoxide↗

Sperm-immobilizing antibodies in women undergoing artificial insemination with donor sperm.

Forty-nine female partners of infertile couples undergoing AID (1 to 44 times) were tested for the presence of sperm-immobilizing antibodies in their serum. One hundred fifty-one infertile women undergoing ART treatment other than AID were enrolled as a control group. One of the 49 AID patients and 3 of the 151 women of the control group were positive for sperm-immobilizing antibodies. The sperm-immobilizing antibodies incidence in AID patients did not differ significantly from that in the control group. This suggests that multiple exposures to semen from different donors do not increase the possibility of inducing sperm-immobilizing antibodies in AID patients.

Antibodies↗

Correlation of human sperm motility characteristics with an in vitro cervical mucus penetration test.

Semen analysis was performed on 226 ejaculates by an integrated microcomputerized system employing the multiple-exposure photography (MEP) method. Mucus penetration tests were performed in vitro using commercial preparations of bovine cervical mucus. A highly significant (P less than 0.001) correlation between mucus penetration distance and sperm count (r = 0.582), motility (r = 0.357), velocity (r = 0.569), motile density (r = 0.582), motility index (r = 0.467), and morphology (r = 0.383) was observed. Increased percentages of immature germ cells (r = -0.318) and bent-tailed sperm (r = -0.221) were the most strongly correlated with mucus penetration. Approximately 10% to 15% of patients with otherwise normal semen parameters displayed poor penetration of mucus. Conversely, 5% to 40% of patients with abnormal semen parameters displayed excellent penetration of the mucus. Motile density and velocity demonstrated the strongest relationship with the outcome of the mucus penetration test. These results suggest that a significant subpopulation of patients can be identified as having inadequate (or adequate) penetration of mucus with otherwise normal (or abnormal) motility characteristics.

Animals↗

Semen analysis data from fresh and cryopreserved donor ejaculates: comparison of cryoprotectants and pregnancy rates.

Patients (155) were selected at random for fresh or cryopreserved semen and inseminated on the predicted day of ovulation. Semen analysis was performed using a microcomputerized multiple-exposure photography system. Frozen semen was used with either glycerol or TEST-yolk (TEST-buffered 20% egg yolk with 10% glycerol) as the cryoprotectant. Cryopreservation resulted in significant decreases in all semen parameters measured. Of these, velocity appeared to be the least effected. TEST-yolk provided significantly more protection against a reduction in velocity compared with glycerol. A total of 18, 17, and 27 patients conceived using fresh, glycerol, or TEST-yolk-preserved semen, respectively. For these same groups, a cumulative pregnancy rate of 52.9%, 27.1%, and 68.5%, respectively, was observed (not significant). The total number of motile sperm per insemination used for fresh artificial inseminations resulting in conception (132.4 X 10(6] was significantly greater than the number used for successful glycerol- and TEST-yolk-preserved semen (approximately 24 X 10(6]. These results demonstrate that although the number of motile sperm of cryopreserved ejaculates are dramatically reduced compared with the fresh counterparts, if a minimum criteria for ejaculate quality is established, the use of cryopreserved semen can offer a viable, effective, and relatively safe alternative to artificial insemination by donor with fresh semen.

Cell Survival↗

Effect of peritoneal fluid on sperm motility and velocity distribution using objective measurements.

The aim of this study was to analyze in vitro the effect(s) of peritoneal fluid (PF) on sperm motility. Seventy PFs obtained during laparoscopy were tested on motile-rich sperm suspensions. Proportion of motile sperm and velocity distribution were evaluated by multiple-exposure photography technique. At time (t) = 0, PFs increased both sperm parameters as compared with control (P less than 0.01). Maximum effect was observed at t = 5 hours: 32 (45%) PFs increased and 5 (7%) decreased the proportion of motile sperm, while 8 (11%) PFs increased and 4 (6%) decreased sperm velocity. No correlation was found between a particular infertile group and a definite negative effect. However, 70% of PFs from fertile women maintained or increased the proportion of motile sperm at t = 5 hours, compared with 36% in the total infertile group. Comparison of the sperm motility effect(s) of a given PF on different ejaculates revealed that the effects observed also were influenced by the sperm sample tested. In conclusion, PFs can maintain or increase the motility of spermatozoa as function of time. However, some PFs can inhibit sperm motility and these effect(s) can be influenced by the sperm sample.

Body Fluids↗

Evaluation and risk factors of latex allergy in spina bifida patients: is it preventable?

Allergic reactions to latex, varying from urticaria to life-threatening IgE-mediated anaphylaxis, have been described in patients with spina bifida. In a study of 50 patients 60% had a latex allergy identified by history, a radioallergosorbent test and/or a skin prick test. Latex allergic patients had undergone statistically more surgical procedures than nonallergic patients (9.5 versus 6.7) with a p value of 0.03. The presence of latex allergy did not correlate with the number of intra-abdominal procedures (1.9 versus 1.4) or with the number of years on clean intermittent catheterization (7.7 versus 5.9). These results suggest that latex allergy is acquired through multiple exposures due to breakdown of blood tissue barriers. Therefore, we recommend that all patients with spina bifida undergoing surgery be evaluated for latex allergy and that all surgical procedures in spina bifida patients, from the time of birth, be performed in a latex-free environment.

Adolescent↗

A critical method of evaluating tests for male infertility.

Considerable uncertainty surrounds the selection of test values that separate infertile from fertile men in the evaluation of male infertility. We herein describe an objective method of determining these values, referred to as threshold values, for different infertility tests. Using test results from fertile men threshold values were chosen such that 96 per cent of the semen samples from the fertile men were scored as fertile. These threshold values then were used to evaluate 100 semen samples from 74 men presenting for evaluation of infertility. Using this method we constructed infertility profiles on each of the 100 semen samples presented for infertility evaluation and found that the zona pellucida-free hamster egg penetration test (a measure of a spermatozoon's ability to undergo capacitation and penetrate an egg) identified 66 per cent of these samples as infertile, while multiple exposure photomicrography (a quantitative measure of sperm motility) identified 54 per cent of these samples as infertile. This compares with results from routine semen analyses using the same method, which identified none of the samples as infertile by sperm motility grade, 1 per cent by semen pH, 4 per cent by the percentage of motile sperm, 7 per cent by the total count of motile sperm, 10 per cent by the total sperm count, 11 per cent by the semen leukocyte concentration, 12 per cent by the concentration of motile sperm, 13 per cent by ejaculate volume, 16 per cent by sperm concentration and 27 per cent by sperm morphology. This method of analyzing infertility test results provides insight into the potential causes of male infertility and offers a critical approach towards understanding the complex problem of male fertility dysfunction.

Female↗