Andrological Parameters in Human Semen of High (≥ 6 ml) and Low (≤ 1 ml) Volume/Andrologische Parameter im menschlichen Samen bei hohem (≥ 6 ml) und niedrigem Ejakulatvolumen (≤ 1 ml)

A study was carried out to evaluate 792 semen of high (greater than or equal to 6 ml) and low (less than or equal to 1 ml) volume and sperm counts ranging from 0.1 to 200 x 10(6) per ml. Particularly emphasized were: motility percentage and grade, percentage viability, morphologically normal sperm and immature cells, and the concentration of fructose in semen (mg/ml). The Duncan multiple range test and the Kruskal-Wallis test with multiple comparison of ranks were used in the statistical analyses. The results show that abnormal semen volume influence neither the basic tendency of andrological parameters to change in accordance with sperm counts of semen nor to correlate with each other. The quality of motility was significantly higher in specimens of high semen volume as compared to those of low semen volume. The concentration of fructose in semen was generally higher in specimens of high volumes than in those of low volumes, including semen devoid of sperm. It was assumed that in general increased semen volume does not affect the quality of other andrological parameters, thus being probably unrelated with fertility potential. The considerably lower values of both motility and viability characterizing the semen of low volume would suggest a lower fertility potential of specimens affected by this type of abnormality.     

Andrological parameters in men with high sperm counts and possible correlation with age

Two hundred eighty six human semen samples with sperm counts above 200 million/ml, were divided into 3 groups according to semen volume: (1) less than 1 ml (low), (2) 1 ml-5 ml (normal), and (3) greater than or equal to 6 ml (high). Each group was examined for routine andrological parameters. Also examined was a possible correlation between sperm counts, semen volumes, and ages of patients. The quality of motility (percent and grade) and viability of sperm were generally lowest in specimens with a volume below 1 ml and highest in those of normal volume. Proportions of morphological normality exhibited a similar trend, lacking, however, statistical significance. The mean fructose content of low-volume semen was significantly decreased in comparison with those of normal volume. Patients with low-volume semen and sperm counts above 200 million/ml were older than those with similar sperm counts and normal volume. The mean ages of all patients with high sperm counts of semen were higher than of those with lower sperm counts. It was assumed that in contradiction to specimens with high sperm counts and low volume, those whose volumes range from 1 ml-5 ml should be considered normal with respect to motility, viability, and morphological normality.     

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.     

Relationship between age and semen parameters in men with normal sperm concentration: analysis of 6022 semen samples

This study evaluates retrospectively the relationship between age and semen parameters among men with normal sperm concentration. It was based on computerized data and performed in an Academic Fertility and IVF Unit. Six thousand and twenty-two semen samples with sperm concentrations of >or=20 x 10(6) ml(-1) were examined according to WHO criteria and analysed in relation to patients' age. For each age group, mean values +/- SD of semen volume, sperm concentration, percentage of motile spermatozoa, normal morphology, acrosome index, total sperm count/ejaculate, total motile sperm count/ejaculate and sexual abstinence duration were examined. A peak semen volume of 3.51 +/- 1.76 ml(-1) was observed at age >or=30 to <35 years and a lowest volume of 2.21 +/- 1.23 ml(-1) was observed at age >or=55 years (P<0.05). Sperm motility was found to be inversely related to age with peak motility of 44.39 +/- 20.69% at age <25 years and lowest motility of 24.76 +/- 18.27% at age >or=55 years (P<0.05). A reduction of 54% was observed for total motile sperm, between values of 103.34 +/- 107 x 10(6) at age >or=30 to <35 years and 46.68 +/- 53.73 x 10(6) (P<0.05) at age >55 years. A statistically significant and inverse relationship was observed between semen volume, sperm quality and patient age, in spite of prolonged sexual abstinence duration. Top sperm parameters were observed at age >or=30 to <35 years, while the most significant reduction in sperm parameters occurred after the age of 55 years.     

Stability of sperm characteristics in men with disturbances in sperm quality

Sixty-one men referred to our laboratory for semen analysis, and subsequently judged to exhibit some form of sperm pathology, were asked to return for a second analysis, not less than 2 months after the first, in order to assess the stability of the pathological changes observed. In almost half of the cases, the referring physician had, on his own initiative, started hormone or antibiotic treatment. The sperm parameters studied included sperm count, sperm motility judged by laser-Doppler spectroscopy, and sperm morphology and viability. The motility characteristics included percentage motile, their average velocity, and percentage swimming in a progressive manner, and their progressive velocity. In untreated subjects, there was no significant difference between the first and second analysis in any of the sperm parameters measured. This was also true for both oligozoospermic individuals (less than 20 x 10(6) sperm/ml) and the group with higher sperm concentrations. All parameters were highly correlated on the two occasions. The average coefficients of variation of the paired observations were highest for sperm count (approximately 25%) and lowest for sperm velocities and the proportion of abnormal and viable cells in the ejaculate (1-9%). No major differences in the extent of variation could be detected between the low and high sperm density groups. In general, the unsystematic antibiotic and hormone regimens (clomiphene or androgen) used by the referring physicians had no discernable effect on any aspect of sperm quality, indicating the need for more controlled and standardized programmes of treatment.     

Seminal factor VII and factor VIIa: supporting evidence for the presence of an active tissue factor-dependent coagulation pathway in human semen

Human semen spontaneously coagulates into a semisolid mass and then wholly liquefies in a process that may have some similarity to that of normal blood. This well described phenomenon is referred to as coagulation and liquefaction of semen. Besides other active components of the haemostatic system, semen contains a significant amount of functional tissue factor (TF). However, TF needs factor (F)VII in order to exert it actions. In this study, we assessed human semen for the presence of FVII and FVIIa, and related their levels to conventional fertility parameters. Using a functional, one stage, clotting assay based upon the prolongation of the prothrombin clotting time, using the ACL 300R analyser and an Imubind FVIIa ELISA assay, FVII and FVIIa levels were measured in 97 semen specimens obtained from sub-fertile (sperm counts <20 x 10(6)/mL), normally fertile (sperm counts >or=20 x 10(6) but <60 x 10(6)/mL), fertile sperm donors (sperm counts >or=60 x 10(6)/mL), vasectomized subjects and in a pooled normal semen parameters group (categorization into groups was based on the World Health Organization guidelines on fertility criteria). In addition, conventional semen parameters were analysed on all semen samples. Both FVII and FVIIa were quantifiable in human semen. The mean levels of FVII and FVIIa were 4.4 IU/dL and 12 ng/mL respectively. Despite the observed variations of FVIIa levels in the studied groups they did not meet statistical significance when the groups were tested against each other. However, seminal FVIIa levels showed a significant positive association with semen liquefaction time, sperm motility and semen volume. The anti-sperm antibodies and sperm-agglutination groups were also associated with raised seminal FVIIa levels. We observed no significant relationship between FVIIa levels and total sperm concentration, sperm count per mL (sperm density), sperm progression and days of sexual abstinence. This study demonstrates that human semen contains appreciable amounts of FVII and FVIIa. It is possible to quantify these using commercially available assays. There also appears to be a direct correlation between the levels of these factors and certain seminal parameters. This finding reinforces the concept of an active clotting system in human semen, by establishing the missing link in the activation of a TF-dependent pathway.     

Semen quality in adult male survivors 5 years after the 2008 Wenchuan earthquake

The influence of the Wenchuan earthquake on semen quality of adult male survivors is unclear. We investigated the semen quality included 673 male survivors from the worse‐affected counties in the earthquake between Aug 2008 and July 2013. Semen parameters including pH, volume, concentration, motility and morphology were measured according to the World Health Organization (WHO) criteria. Kruskal–Wallis analysis of variance was used to examine the statistical differences between years, and a logistic regression was used to analyse the impacts caused by earthquake on the changes of semen quality. We found the medians (5th and 95th) were 2.5 ml (0.6–5.5) for semen volume, 59.0 × 10⁶ ml^?1 [(13.0–133.0)] × 10⁶ ml^?1 for semen concentration, 46% (13–64%) for sperm progressive motility and 3.0% (0–17.5%) for normal morphology for adult male survivors. Semen concentration, the percentage of sperm progressive motility, total motility and sperm normal morphology were all decreased in the first 3 years, and the differences among years 1, 2 and 3 were significant except the percentage of sperm progressive motility (P < 0.05). The casualties and heavy housing damage caused by earthquake had a negative effect on semen quality. The main findings will provide further diagnosis and therapy basis of male fertility by data, for affected populations in the earthquake.     

Secular and seasonal changes in semen quality among young Danish men: a statistical analysis of semen samples from 1927 donor candidates during 1977-1995

The objective of this study was to investigate whether semen quality has changed during the years 1977-1995 in a group of unselected semen donor candidates, and to determine whether semen quality is subject to seasonal variation, by analysis of time- and season-related changes in semen quality using multiple regression and ANOVA. The study was based on analysis of the first semen sample delivered by 1927 semen donor candidates in Copenhagen during the period 1977-1995, with determination of semen volume, sperm concentration, total sperm count, percentage motile spermatozoa, and a semiquantitative sperm motility score. Multiple linear regression analysis with year, sexual abstinence and season as covariates showed a significant increase in mean sperm concentration from 53.0 x 10(6)/mL in 1977 to 72.7 x 10(6)/mL in 1995 (p < 0.0001) and in mean total sperm count from 166.0 x 10(6) to 227.6 x 10(6) (p < 0.0001). Mean semen volume and percentage motile spermatozoa did not change. Sperm motility deteriorated, as the spermatozoa in 74.2% of the samples were of excellent motility in 1977-1980 compared to only 41.9% in 1993-1995 (chi 2 = 130.0, p < 0.0001). Analysis of variance showed significant variation between seasons regarding sperm concentration (p < 0.0001) and total sperm count (p < 0.0001). Highest sperm counts were found in spring, with a mean concentration (95% C.I.) of 77.6 x 10(6)/mL (71.9-83.7), and lowest in summer, with a mean of 57.5 x 10(6)/mL (50.1-65.4). No other semen parameter varied with season. It is concluded that sperm counts increased, whereas sperm motility decreased, in a group of Danish semen donor candidates, from 1977 to 1995. Due to the retrospective design and the anonymity of the donors, we were unable to control for variation in donor age, and we cannot exclude the possibility that some donor candidates were selected by being accepted as donors by other semen donor services in Copenhagen. With these limitations in mind, we suggest our results should be interpreted cautiously and regarded as a contribution to the ongoing dispute on whether or not there is a continuous decrease in sperm quality. The seasonal variations found in sperm concentration and total sperm count were pronounced and were not attributable to seasonal differences in the length of sexual abstinence. Additionally, the same seasonal pattern was observed in five successive year-intervals. These findings strongly indicate that human testicular function is influenced by season, a phenomenon well known in many lower mammals.     

Oligozoospermia: recent prognosis and the outcome of 73 pregnancies in oligozoospermic couples

The minimum value for each of the five main semen parameters, below which conception rarely occurred or did not occur at all, was calculated in a group of 1884 couples complaining of primary and secondary infertility: 304 conceptions including first as well as consecutive conceptions, occurred. The parameters evaluated were (minimum value calculated in this study between brackets) volume (1.0 ml), sperm count ml(-1) (2.0 million), total sperm count (4.0 million), motility (10%), forward progression (2.0 MacLeod units: scale 1-4) and normal sperm morphology (3%). The pregnancy rate in the group of 308 oligozoospermic men and the minimum value of semen parameters were the cornerstones in determining the prognosis for oligozoospermic patients. A sperm count of >2.0 million ml(-1) was considered relatively adequate for eventual conception judged by the 68 of 308 (22.1%) pregnancies that occurred among oligozoospermic men in this study, provided that the other five semen parameters showed values above the minimum value. In cases where the average sperm count was <2 million ml(-1), the chances for conception became rare, viz five of 308 (1.6%). The total number of pregnancies in the group classified as oligozoospermic was 73 (23.7%). With these pregnancies there was no increase in the rate of foetal wastage and congenital abnormalities. Abortion occurred in 15.09% and ectopic pregnancy in 0.9% among first and consecutive pregnancies. One infant among the 56% boys and 44% girls was born with congenital abnormalities. Most of these infants had a normal birth mass of >2500 g.     

The reference values for semen parameters of 1213 fertile men in Guangdong Province in China

Semen samples were collected from 1213 fertile men whose partners had a time-to-pregnancy (TTP) ≤12 months in Guangdong Province in Southern China, and semen parameters including semen volume, sperm concentration, total counts, motility, and morphology were evaluated according to the World Health Organization (WHO) 2010 guideline. All semen parameters analyzed were normal in ~62.2% of the total samples, whereas ~37.8% showed at least one of the semen parameters below normal threshold values. The fifth centiles (with 95% confidence intervals) were 1.3 (1.2–1.5) ml for semen volume, 20 × 10⁶ (18×10⁶–20×10⁶) ml⁻¹ for sperm concentration, 40 × 10⁶ (38×10⁶–44×10⁶) per ejaculate for total sperm counts, 48% (47%–53%) for vitality, 39% (36%–43%) for total motility, 25% (23%–27%) for sperm progressive motility, 5.0% (4%–5%) for normal morphology. The pH values ranged from 7.2 to 8.0 with the mean ± standard deviation at 7.32 ± 0.17. No effects of age and body mass index were found on semen parameters. Occupation, smoking and alcohol abuse, varicocele appeared to decrease semen quality. Sperm concentration, but not sperm morphology, is positively correlated with TTP, whereas vitality is negatively correlated with TTP. Our study provides the latest reference values for the semen parameters of Chinese fertile men in Guangdong Province, which are close to those described in the new WHO guidelines (5^th Edition).     

Sialyl transferase in human semen

The activity of the enzyme sialyl transferase, which is responsible for the transfer of sialic acid, has been examined in the following human seminal plasma: (1) originating from semen with sperm counts above 30 X 10(6)/ml and exhibiting normal values of other andrological parameters; (2) from semen with sperm counts below 25 X 10(6)/ml and with other andrological parameters of poor quality; (3) from semen devoid of sperm. Additionally, enzyme activity was examined in fractionated (split) semen. The activity in plasma based on measurement of radioactivity incorporated into asialofetuin, following incubation in the presence of 14C sialic acid, was expressed as cpm both per volume and per mg protein. There was a tendency for decreased activity with the decrease in sperm density of semen. The difference, however, lacked statistical significance. Enzyme activity in split semen was significantly lower in the first than in the second fraction.     

尿激酶对弱精症患者精子活力影响的研究

对２８例弱精症男子精液中加入不同浓度的尿激酶后，分别在３０、６０、９０和１２０分钟观察精子的活率和前向运动能力。并对１４例患者体内给予尿激酶１０天后，比较用药前后精子活率和有前向运动力精子数目的变化。结果：２８例精液标本体外加入尿激酶后，３０、６０、９０和１２０分钟时精子的活率均有提高；前向运动精子的数目亦有增加。体内给药前，１４例精子平均活率为４４．６４±１１．７９，用药１０天后，精子平均活率为６４．７１±１３．０１，二者比较有极显著差异（Ｐ＜０．０１）。尿激酶的这一作用可能与它降低精液的粘稠度有关。     

Separation of sub-populations of sperm with higher fertility potential from normal and pathological semen by peanut agglutinin

Incubation of human sperm from semen with counts below 20 x 10(6)/ml with peanut agglutinin (PNA) resulted in agglutination of about 70% of sperm. An inverse correlation was found between non-agglutinating (PNA) sperm and sperm density (r = 0.48, p less than 0.01) and a direct correlation with acrosome-damaged sperm (r = 0.83, p less than 0.001). Binding of 125I-PNA to sperm revealed high, and possibly also low affinity binding sites on sperm from both normozoospermic and oligozoospermic origins. Quantitative transmission electron microscopy analysis revealed that agglutinated, PNA+, sperm had lower frequency of acrosomeless spermatozoa than PNA(-)-sperm (44% versus 77%) and higher fertility score (+0.18 +/- 0.1 and -1.5 +/- 0.87 respectively, p less than 0.01). Removal of PNA- sperm from oligozoospermic semen may increase the fertility score of the remaining sperm.     

Beta-endorphin and calcitonin in human semen

The levels of beta-endorphin (beta-E) and calcitonin were estimated in 36 samples of seminal plasma from semen of normospermic, oligozoospermic, and azoospermic origins and in pools of isolated sperm. The mean levels in plasma calculated for all samples examined were 192 +/- 224 pg/ml for beta-E and 754 +/- 397 pg/ml for calcitonin. The amounts in sperm were as follows: for beta-E in pools with sperm counts of 0.1-10 x 10(6)/ml, 157.2 +/- 99.7 pg/10(8) and 27.9 +/- 23.6 pg/ml protein; in pools of greater than 10-30 x 10(6)/ml, 71.2 +/- 41.5 pg/10(8) and 6.5 +/- 1.2 pg/mg protein; in pools of greater than 30-200 x 10(6)/ml, 24.9 +/- 9.7 pg/10(8) and 61 +/- 1.9 pg/mg protein. For calcitonin the amounts were: 501.2 +/- 170.8 pg/10(8) and 27.4 +/- 21.5 pg/mg protein, correspondingly. It was suggested that beta-E and calcitonin present in seminal plasma are synthesized mostly in a compartment of the male reproductive system. The high cellular beta-E and calcitonin levels would be involved in the process of motility through their effect on calcium transport.     

Semen quality changes among 2343 healthy Slovenian men included in an IVF-ET programme from 1983 to 1996

To determine whether semen quality in Slovenians has changed over 14 years (1983-96), we analysed retrospectively the semen of 2343 healthy men with a normal spermiogram, who were partners of women with tubal infertility included in the IVF-ET programme. Age at semen collection, duration of sexual abstinence, semen volume, sperm concentration, total sperm count, percentage of spermatozoa with progressive motility, and normal morphology were determined. Multiple regression analysis was used to assess the changes in sperm characteristics according to the year of semen collection, year of the man's birth and the duration of sexual abstinence. Semen volume, sperm concentration, sperm count and total sperm motility did not change between 1983 and 1996, whereas between 1988 and 1996 rapid progressive sperm motility decreased by 0.95% per year (p < 0.0001). Semen volume, sperm concentration, and sperm count increased with duration of sexual abstinence. After adjustment for the year of semen collection and duration of sexual abstinence, multiple regression analysis showed that sperm concentration decreased by 0.67% per each successive year of birth (p = 0.03). Thus the sperm concentration decreased from 87.6 x 10(6)/mL in men born in the 1940s to 77.3 x 10(6)/mL in those born between 1956 and 1960. After 1960, sperm concentration was found to increase. In 2343 healthy men, no decline in semen quality, except in rapid progressive motility, was observed in the study period. Lower sperm concentration was found among men born between 1950 and 1960. This could be related to worse socio-economic status, stress or negative environmental factors in this time period.     

Semen quality evaluation in a cohort of 28213 adult males from Sichuan area of south‐west China

The trends in semen quality are conflicting. Although many previous surveys on semen quality indicated a decline, the trends in semen quality in Sichuan area of south‐west China are not clear. We analysed the semen parameters in a cohort of 28 213 adult males close to general population in Sichuan between July 2007 and June 2012, and investigated the changes on semen quality. The semen parameters including pH, volume, concentration, motility, morphology were measured according to the World Health Organization (WHO) criteria. Kruskal–Wallis analysis of variance was used to examine the statistical differences of semen quality between groups. We found that the medians (5th and 95th percentiles) were 2.4 ml (1.0–5.0) for semen volume, 62.0 × 10⁶ ml^?1 (15.0–142.0) for semen concentration, 39% (18–60%) for sperm progressive motility and 10.5% (1.0–34.5%) for normal morphology. In these 5 years, sperm concentration and the percentage of sperm normal morphology were decreased from 66.0 × 10⁶ ml^?1 to 49.0 × 10⁶ ml^?1 and from 13.5% to 4.5%, respectively; among different reproductive history groups, sperm concentration and the percentage of sperm normal morphology were also decreased in these 5 years. And the incidence of azoospermia was increasing. These may imply that there is a decline in semen quality of adult males in Sichuan area.     

Adenosine triphosphate levels in human spermatozoa

ATP levels in human spermatozoa have been positively correlated with good motility. This has given rise to the impression that good ATP levels per se are related to good motility, i.e., the higher the ATP level, the better the motility and fertilizing potential. There was no direct correlation between motility percentage, forward progression, viability percentage, and ATP levels (when expressed per 1 million spermatozoa) in the general population. This finding was not unexpected since other factors, such as defects of the microtubules and viscosity of the semen, could affect the motility in some patients. However, when semen from individual patients was assessed, the motility percent, viability, and ATP concentration decreased by comparable levels over a 4-h period postejaculation. Semen samples with normal counts of spermatozoa (greater than 20 X 10(6)/ml) had higher levels of ATP than samples from patients with oligozoospermia. Spermatozoa from patients whose semen contained less than 0.60 X 10(-2) nmol ATP/1 million spermatozoa showed a rapid drop in motility over a 4-h period compared with semen samples where the motility remained above 10% motile over this period, the latter samples having ATP levels averaging 3.30 X 10(-2) nmol/1 million spermatozoa.     

Factors affecting the variability of semen analysis results in infertile men

Infertile men who had 3 or more semen analyses performed in one laboratory were placed in 2 groups (I) oligozoospermic group (n = 106), mean sperm concentration between 1 and 20 million/ml (II) asthenozoospermic group (n = 71), mean sperm concentration greater than 20 million/ml, and mean motility less than 60%. With increasing durations of abstinece from ejaculation before the tests there were significant increases in semen volume and sperm concentration. Semen volume increased over the first 4 days to a similar extent in both groups. Sperm concentrations increased over 15 days, but the effect of abstinence was much greater in the asthenozoospermic group than in the oligozoospermic group (14% compared with 1.4% of the within subject variation). Significant changes in results accompanied repeated testing, notably rises in sperm concentration and motility. Sperm motility was lower in winter and higher in summer in both groups and also, but to a lesser extent, in artificial insemination donors who collected semen in the laboratory.     

Concentrating human sperm before cryopreservation

To improve the quality of cryo-preserved sperm, ejaculated semen was concentrated prior to freezing by means of continuous-step density gradient centrifugation. Freezing was simplified by employing liquid nitrogen vapor with KS-II cryo-medium. The original semen (n = 32, 41 +/- 13 x 10(6)/ml, 17 +/- 11% motility) was improved by the processing to be 68 +/- 29 x 10(6)/ml, 55 +/- 19% motility. Even after the specimens were thawed, sperm concentrations were similar to those of the original semen and 39 +/- 18% motility remained with a mean survival rate of 74 +/- 7.7%. The improved sperm motility and higher survival rate resulted in better post-thaw sperm quality than those of the original semen.     

精子优选后冷冻保存对活性氧生成的影响

目的观察优选技术在精液冷冻保存中对常规精液参数、线粒体膜电位(△Ψm)和活性氧(ROS)的影响。方法分别用计算机辅助精子分析仪(CASA)、流式细胞仪(FCM)检测优选或未优选的精子在冷冻前后常规精液参数、△Ψm和精子细胞内ROS的变化。结果各冻融组精子活率、a级精子、(a+b)级精子、正常形态率均比冷冻前有不同程度降低,有显著性差异;而优选冻融组和优选加精浆冻融组间常规精液参数无显著性差异(P>0.05),但都明显高于原始冻融组(P<0.01)。原始精液冷冻后比冷冻前△Ψm下降(P<0.05),ROS显著上升(P<0.01)。优选冻融组和优选加精浆冻融组与冷冻前相比,以及组间无显著性差异(P>0.05);但与原始冻融组分别比较,△Ψm均显著提高(P<0.05),ROS均显著减少(P<0.01)。各冻融组△Ψm和ROS都有明显相关性,相关系数(r)依次为-0.528、-0.537、-0.606(P<0.001)。结论精液优选后冷冻可以在保持良好的△Ψm同时,减少ROS生成,因此建议临床可应用该技术冷冻保存精子。