Toxic Environmental Chemicals in Human Semen: Analytical Method and Case Studies/Toxische Umweltchemikalien in menschlichem Sperma: Analysenmethode und Fallstudien.

In 1982, 89 men from all over the FRG participated in an investigation of sperm density and the presence of selected persistent environmental chemicals in their semen. Semen concentrations for the following chemicals were measured: lead, cadmium, hexachlorobenzene (HCBC), a-hexachlorocyclohexane (a-BHC), DDT and metabolites, dieldrin and polychlorobiphenyls (PBC). Heavy metal analyses were performed by Zeeman-AAS with direct sample measurement. A method for the quantitative determination of chlorinated hydrocarbons in semen by liquid-liquid extraction, clean-up and quantitation by gas chromatography with an electron capture detector. Recovery rates ranged from 72 to 120%. Compound identification in samples was confirmed by negative chemical ionization mass spectrometry. Heavy metals and organochlorine compounds in semen were present in the same concentration range as in blood or other biofluids. Occupational exposure as well as extraordinary environmental exposure appeared to influence semen concentrations. Students in chemistry showed elevated levels of organochlorine compounds. Men who lived in the vicinity of an atmospheric source of heavy metals showed strikingly the vicinity of an atmospheric source of heavy metals showed strikingly elevated levels of semen lead and cadmium. We were unable to find a statistically significant correlation between sperm density and any of the variables examined in this study including: orgonochlorine compounds summed or individually; lead and/or cadmium; and tobacco consumption. Significant correlations were found between the simultaneous occurrence of lead and cadmium and between the simultaneous occurrence of HCB, DDT and DDE.     

Blood concentrations of lead, cadmium, mercury, zinc, and copper and human semen parameters.

The study consisted of 35 male subjects attending an andrology clinic. The subjects all had poor sperm parameters that could not be attributed to any known medical cause. The objective was to evaluate the relation between various seminal characteristics (volume, total sperm count, sperm viability, proportion of progressively motile sperm, and different sperm morphology) and the blood concentrations of lead, cadmium, mercury, copper, and zinc. The mean blood concentrations of lead, mercury, copper, and zinc were within the normal values; cadmium concentration (1.35 micrograms/L) was much higher than the norms. Asthenozoospermic subjects had significantly (p less than .025) higher blood cadmium levels than normozoospermic subjects. No significant differences were noted between the two groups for mean concentration of mercury, zinc, and copper in blood. Significant correlations were observed between blood cadmium levels and volume of semen, midpiece defects, and immature forms of spermatozoa. High blood cadmium levels may have an effect on spermatogenesis. Possible reasons for the high blood cadmium levels among the subjects are discussed.     

非职业接触人群中不育夫妇卵泡液及精液中4种重金属元素的含量测定

目的:探讨非职业接触人群中不育夫妇卵泡液和精液中4种重金属:铅、镉、锌和锰的含量。方法:女性行IVF-ET操作,常规取卵后收集卵泡液,丈夫手淫法留取精液,用火焰原子吸收法测定卵泡液及精液中的锌含量,用石墨炉原子吸收法测定铅、镉和锰含量。结果:铅、镉、锌和锰的测定均值在卵泡液中分别是(151.06±421.03)μg/L、(2.02±2.79)μg/L、(0.54±0.55)mg/L和(28.54±50.90)μg/L,在精液中分别是(250.23±600.88)μg/L、(7.42±15.84)μg/L、(189.11±283.08)mg/L和(82.15±202.65)μg/L;检出率在卵泡液中分别为43.8%(21/48)、22.9%(11/48)、75.0%(36/48)和50.0%(24/48),在精液中分别为70.2%(33/47)、31.9%(15/47)、100.0%(47/47)和46.8%(22/47)。结论:精液中4种重金属元素含量均高于卵泡液,检出率除锰以外,也均以精液为高。     

Cadmium, lead, selenium, and zinc in semen of occupationally unexposed men

Summary. Concentrations of cadmium, lead, selenium, and zinc were determined in semen and seminal plasma of 22 volunteers by atomic absorption spectrometry. Additionally conventional semen parameters and, by means of computer videomicrography, motion parameters of spermatozoa were evaluated. Concentrations of Cd, Pb, and Zn determined in semen were not significantly different from those measured in seminal plasma. However, selenium levels were significantly higher in semen (53.8 ± 22.9 μg 1⁻¹) than in seminal plasma (40.4 ± 15.5 μg 1⁻¹, P <0.01). The investigated semen samples on average contained low levels of Cd (0.4 ± 0.23 μg 1⁻¹) and Pb (9.8 ± 6.5 μg 1⁻¹). Studies on the intra-individual variability revealed the following average coefficients of variation (%) for element concentrations: Pb (70), Cd (53), Se (27), and Zn (23); and for semen parameters: total sperm count (46), sperm concentration (37), motility (22), ejaculate volume (21), linearity (19), linear velocity (11), curvilinear velocity (10), and percentage of normally formed sperm (9). Significant positive correlations were detected between semen selenium levels and sperm concentration ( r =0.51, P <0.05), and percentage of normally formed sperm ( r =0.46, P <0.05), respectively. Sperm motility ( r =0.53, P <0.02), linear ( r = 0.76, P <0.001) and curvilinear velocity ( r = 0.64, P < 0.002) were significantly correlated with semen cadmium levels.     

Content of chlorinated pesticides in human semen of a random population

Residues of chlorinated pesticides - BHC isomers, DDT isomers, HCB and DMDT - in samples of human semen have been determined. Samples of semen were collected from a random population of patients sent to our department for the evaluation of their fertilizing ability. The contents of 50 samples of semen were analyzed individually for residues of chlorinated pesticides and their metabolites by gas-liquid chromatography. Of BHC isomers detected in the analyzed samples of semen, the highest absolute values were shown for epsilon-BHC (x 0.005 μg/g of sample). DMDT, op'DDT and pp'DDT were not detected in the studied samples. DDE was found in 20 samples on the average level of 0.003 μg/g.     

Correlation between lead and cadmium concentration and semen quality

There are contrary reports of association of lead and cadmium with the decline in semen quality. This study evaluates whether seminal lead (Pb) and cadmium (Cd) at environmental concentration are associated with altered semen quality. We conducted a study of healthy fertile and infertile men 20–43 years of age attending the Andrology Laboratory of Reproductive Biology Department for semen analysis. The semen analysis was carried out according to the WHO 2010 guidelines. Seminal lead and cadmium were estimated by ICP‐AES. The lead and cadmium values were significantly higher in infertile subjects. A negative association between seminal lead or cadmium concentration and sperm concentration, sperm motility and per cent abnormal spermatozoa was found. This study shows that exposure to Pb (5.29–7.25 μg dl^?1) and cadmium (4.07–5.92 μg dl^?1) might affect semen profile in men. Age, diet, smoking and tobacco chewing habits may have an influence on the increase in exposure to Pb and Cd in the individual subjects.     

Seminal lead and copper in fertile and infertile men: Blei und Kupfer im Spermaplasma bei fertilen und infertilen Männern

Lead and copper concentrations were determined by atomic absorption spectroscopy in semen from 18 fertile and 172 infertile men. Significant correlations between copper concentrations in semen and sperm concentration (r = 0.32, P less than 0.001), percentage progressive motility (r = 0.23, P less than 0.005) and normal morphology (r = 0.22, P less than 0.005) were observed, while no such correlation existed for lead. However, semen copper concentrations of infertile men (194.99 +/- 5.70 micrograms l-1) and fertile men (183.39 +/- 14.37 micrograms l-1) did not differ significantly. Mean lead concentration in semen of fertile men was 11.18 +/- 0.62 micrograms l-1 and significantly higher than lead concentration in semen of fertile men (5.61 +/- 0.53 micrograms l-1, P less than 0.006). Reinvestigation of 18 infertile men after 2 years showed a significant drop of lead concentrations in semen from 17.31 +/- 1.41 to 6.94 +/- 1.32 micrograms l-1 (P less than 0.0002), which might be related to the decreasing use of leaded petrol in the Federal Republic of Germany.     

邻苯二甲酸二酯与特发性弱精子症的关系

目的 检测人精浆中邻苯二甲酸二酯(DEHP)的浓度来评价DEHP对特发性弱精子患者精液质量的影响.方法 选择特发性弱精子患者50例,非特发性弱精子患者50例,精液正常者50例,分别用非特发性弱精子患者组及精液正常者组作为对照组.运用反相液相高效色谱法测定血浆及精浆中DEHP浓度.并运用SPSS分析软件测定结果进行了分析.结果 特发性弱精子患者组中DEHP均可检出,含量范围在精液中为ND0.10-1.5 mg/L;非特发性弱精子患者组及精液正常组DEHP分别检出15例及13例,精液中为ND0～0.96 mg/L和ND0～0.95mg/L;特发性弱精子组与非特发性弱精子组和正常组精浆中DEHP的浓度有差异(P＜0.05),非特发性弱精子症患者组与正常组精浆中DEHP浓度无差异(p＞0.05),精浆中DEHP浓度与精子活力负相关(p＜0.05).结论 精浆中DEHP可以影响精子质量.

Abstract：

Objectives To detect the level of di-2 - ethylhexyl phthalate(DEHP) in serum and semen to evaluate effect of DEHP to idiopathic asthenaspermia. Methods 150 people were assigned into idiopathic asthenaspermia group( n = 50), no idiopathic asthenaspermia group ( n = 50 ) and nomal group ( n = 50 ). The concentrations of DEHP in the human biological samples were measured by using reversed-phase HPLC. Results The DEHP were detected in most of the idiopathic asthenaspermia group 0. 10 ～ 1. 5 mg / L in semen specimens, 15 of no idiopathic asthenaspermia group 0 ～0.96 mg/L in semen specimens, 13 of nomal group 0 ～0.95mg/L in semen specimens. There was difference between semen concentrations of DEHP and sperm molity ( p ＜ 0. 05 ). Conclusions These results suggest that DEHP affect semen quanlity,specialy DEHP in semen specimens.     

Antibacterial skin preparation decreases the incidence of false-positive semen culture results

PURPOSE: Antibacterial skin preparation was used before obtaining semen cultures to assess whether true seminal pathogens could be isolated with greater specificity. MATERIALS AND METHODS: A total of 14 paired semen cultures were obtained from men evaluated for infertility. The first set (group 1) was obtained without preparation and the second set (group 2) was obtained after using an antibacterial skin preparation consisting of a shower followed by perineal, penile and hand wash with 4% chlorhexidine and 10% povidone-iodine. The sets were compared and statistical analysis was performed using the paired t test. RESULTS: In group 1, 25 total bacterial isolates were detected compared to 13 in group 2, which was statistically significant (p <0.002). In group 1, 11 enteric organisms were isolated compared to 5 in group 2, which was also statistically significant (p <0.05). These data suggest that the source of most enteric organisms isolated from semen is genital and perineal skin. CONCLUSIONS: A significant reduction in enteric contaminants, which would otherwise be erroneously treated, can be achieved with an effective antibacterial skin preparation. We were able to isolate true pathogens with greater specificity with (80%) than without (63%) an antibacterial skin preparation. An antibacterial skin preparation reduces false-positive cultures by at least 50%, which would lead to a significant reduction in costs and morbidity.     

Usefulness of sperm quality analyzer-V (SQA-V) for the assessment of sperm quality in infertile men

The aim of this study was to evaluate the correlation of new of Sperm Quality Analyzer (SQA-V) with the computer-assisted sperm analysis (CASA) and manual semen analysis estimates. One hundred five fresh semen samples were analyzed using SQA-V and CASA and manual semen analysis. Significant correlations of sperm concentration (p < 0.0001), sperm motility (p < 0.0001), and normal morphology (p < 0.0001) were observed between SQA-V variables and manual semen analysis estimates. There also were significant correlations of sperm concentration (p < 0.0001), sperm motility (p < 0.0001), and sperm velocity (p = 0.0235) between SQA-V variables and CASA estimates. Meanwhile, it did not correlate with amplitude of lateral head displacement, beat cross frequency, lineality assessed by CASA. The value of the sperm concentration and the sperm motility measured by SQA-V showed high correlations with the value of those measured by CASA and manual semen analysis. In addition, velocity and sperm morphology may also be evaluated to some extent using SQA-V.     

Seminal Factor VIII and von Willebrand Factor: a possible role of the conventional clotting system in human semen?

Factor (F) VIII circulates in blood complexed with von Willebrand Factor (vWF). Deficiency or defect accounts for haemophilia A and vWF disease. In blood, FVIII functions as a co-factor for FIXa in the activation of FX. Human semen coagulates and liquefies in a process that resembles and has some links with the conventional haemostatic process. A study elsewhere has detected traces, but not measurable levels, of FVIII coagulant activity (FVIII:C). In the present study we have assessed FVIII antigen (FVIII:Ag), FVIII:C and vWF antigen (vWF:Ag) levels in 159 semen specimens obtained from sub-fertile (n = 21), normally fertile (n = 38), fertile donors (n = 32), and vasectomized men (n = 57). Seminal FVIII:Ag levels were also measured in a group defined by several parameters derived from the World Health Organization (WHO) fertility criteria, termed "pooled normal semen parameters" (PNSP). Factor VIII:Ag levels were compared with conventional fertility parameters. In addition, both FVIII:C and vWF:Ag were assessed in a separate group of normal individuals (n = 11). Factor VIII:Ag, FVIII:C and vWF were present and quantifiable in human semen. Factor VIII:Ag levels were significantly lower in vasectomy subjects compared with donors (p = 0.01) or PNSP group (p = 0.01). Several trends taken together suggest an associations between FVIII:Ag and semen quality. Parallel investigations demonstrate FV, FVII, FVIIa, FIX, FIXa, FXa, FXI, FXII, tissue factor (TF) and tissue factor pathway inhibitor (TFPI) in semen. The present report therefore provides further evidence for the presence of a functioning clotting system in human semen.     

Human Seminal Plasma Cadmium: Comparison with Fertility and Smoking Habits/Cadmium in menschlichem Seminalplasma: Vergleich zwischen Fertilität und Rauchgewohnheiten

Cadmium, selenium and zinc were determined in seminal plasma and serum of 64 men by atomic absorption spectrometry (AAS). The mean (+/- SD) cadmium concentrations in seminal plasma and serum were 0.22 +/- 0.22 micrograms and 0.28 +/- 0.10 micrograms, respectively, but they did not correlate with each other. Smokers (n = 31) had significantly (p less than 0.01) higher serum cadmium concentrations than non-smokers (n = 31). Also seminal plasma cadmium in smokers was elevated, but a significant difference to non-smokers was only found if more than 20 cigarettes were consumed daily. No differences were found in semen quality and fertility between smokers and non-smokers. The seminal plasma cadmium had no correlation to selenium or zinc which, however, displayed a positive correlation (r = 0.852, p less than 0.001) to each other. It is concluded that smoking increases the exposure to cadmium. Although no obvious reproductive suppression was observed, heavy smoking may possibly enhance toxic effects in men under other detrimental exposures.     

Comparison of the Content of Chlorinated Pesticide Residues in Human Semen, Testicles and Fat Tissues

Studies were undertaken to determine the residues of chlorinated pesticides in human semen, testicles and fat tissues. Samples of semen were taken from a random population and samples of tissues at surgery. A preliminary estimation was made to assess the level gradient of environmental pollutants in these types of specimens characterized by a different fat content. Pesticides were considered to show an affinity to fat. The results are an early stage in a programme for a detailed analysis of the effect of pesticides on human reproductive ability and on the value of genetic material.     

Sperm chromatin heterogeneity as an infertility factor

Semen samples from husbands with a history of unexplained infertility (n = 33), of women with habitual abortion (n = 36), or normal fertile donors (n = 20) were subjected to conventional semen analysis (SA), Acridine orange test (AOT), and zona-free hamster egg penetration test (HEPT). The three tests operate independently. The most discriminatory test was AOT (p = 0.0001) followed by HEPT (p = 0.019). The frequency of sperm chromatin heterogeneity as detected by AOT red fluorescence was highest in habitual abortion (39.4%), followed by unexplained infertility (16.4%), and, last, donors (9.4%). However the percentage of penetration was highest in habitual abortion (50.7%), followed by donors (43.1%), and least in unexplained infertility (33.9%). Conventional semen parameters (sperm density, motility, abnormality, and vitality) were the least to discriminate between the three groups. The presence of abnormal sperm chromatin may lead to infertility as a result of early pregnancy loss.     

Semen quality in fertile US men in relation to geographical area and pesticide exposure

We conducted the first US study to compare semen quality among study centres using standardized methods and strict quality control. We present data on semen quality in partners of 493 pregnant women recruited through prenatal clinics in four US cities during 1999-2001. Sperm concentration, semen volume and motility were determined at the centres and morphology was assessed at a central laboratory. While between-centre differences in sperm morphology and sample volume were small, sperm concentration and motility were significantly reduced in Columbia, MO (MO) relative to men in New York, NY, Minneapolis, MN and Los Angeles, CA; total number of motile sperm was 113 x 10(6) in MO and 162, 201 and 196 x 10(6) in CA, MN and NY respectively. Differences among centres remained significant in multivariate models that controlled for abstinence time, semen analysis time, age, race, smoking, history of sexually transmitted disease and recent fever (all p-values <0.01). We hypothesized that poorer sperm concentration and motility in MO men relative to other centres might be related to agricultural pesticides that are commonly used in the mid-west. We investigated this hypothesis by conducting a nested case-control study within the MO cohort. We selected 25 men in this cohort for whom all semen parameters (concentration, % normal morphology and % motile) were low as cases and an equal number of men for whom all semen parameters were within normal limits as controls. We measured metabolites of eight non-persistent, current-use pesticides in urine samples the men had provided at the time of semen collection. Pesticide metabolite levels were elevated in cases compared with controls for the herbicides alachlor and atrazine, and for the insecticide diazinon (2-isopropoxy-4-methyl-pyrimidinol) (p-values for Wilcoxon rank test = 0.0007, 0.012, and 0.0004 for alachlor, atrazine and diazinon respectively). Men with higher levels of alachlor or diazinon were significantly more likely to be cases than men with low levels [odds ratios (OR) = 30.0, 16.7 for alachlor and diazinon respectively], as were men with atrazine over the limit of detection (OR = 11.3). These associations between current-use pesticides and reduced semen quality suggest that agricultural chemicals may have contributed to the reduced semen quality seen in fertile men from mid-Missouri.     

Time-related decline in sperm motility patterns in men with cytotoxic antibodies

Sperm motility patterns in semen from 10 fertile nonautoimmune men (fertile control group) and 33 infertile men with various titers of cytotoxic sperm antibodies in their seminal plasma (group 1: titers less than or equal to 16, n = 6; group 2: titers 64 to 512, n = 12; group 3: titers greater than or equal to 1024, n = 15) were evaluated every 2 hours for 12 hours and finally at 24 hours. A significant decline in sperm swimming speed and linearity was observed at 6 hours in semen from 27 infertile men with sperm antibodies. Beginning at 8 hours, semen from sperm antibody-positive men in group 2 showed a significant decline in percentage motile sperm (p less than 0.001) compared to the fertile controls. The percentage motility in semen of donors in groups 1 and 3 was significantly lower than that in semen of fertile donors at 10 hours (p less than 0.05), 12 hours (p less than 0.01), and 24 hours (p less than 0.001). The mean velocity in groups 2 and 3 was significantly less than that in fertile controls at 10 and 12 hours (p less than 0.05). The linearity of sperm motility started to decline 4 hours after semen samples were obtained from sperm antibody-positive groups 2 and 3 in contrast to sperm antibody-negative fertile or infertile groups (p less than 0.05). It is concluded that the presence of cytotoxic sperm antibodies in the seminal plasma adversely affects sperm linearity as early as 4 hours after semen collection.(ABSTRACT TRUNCATED AT 250 WORDS)     

内分泌干扰物对精液质量的影响

内分泌干扰化合物(endocrine disrupting chemicals,EDCs)是指外源性的能干扰人类或动物内分泌系统诸环节并导致异常效应,或使其后代内分泌功能发生改变的化学物质。EDCs种类繁多,广泛存在于土壤、水、甚至食物中。本文从农药、塑料增塑剂和洗涤剂这3类常见EDCs着手,从流行病学和生物学角度阐述了其对人类及动物精液质量的影响。     

Semen levels of mitochondrial DNA deletions in men attending an infertility clinic do not correlate with phenotype

In view of previous reports associating mitochondrial DNA deletions with male reproductive disorders, levels of the `common' 4977 bp mitochondrial DNA deletion were evaluated semi-quantitatively in 64 men, without prior knowledge of the clinical diagnosis. Significant levels of deletions were detected in 34/64 men (53%) but 29 of these (45%) had a normal semen profile and were phenotypically normal. No deletions were detected in 30 men, of whom 21 were normospermic, six were oligozoospermic and three were azoospermic. It is concluded that although mitochondrial DNA deletions within the testis may be associated with primary testicular disease, no correlation with semen quality was evident in this study, thus limiting its potential use as a diagnostic test.     

精液凝固及粘稠度异常的区别

目的采用显微镜观察和计算机辅助的精子运动分析对人类精液凝固-液化和粘稠度异常进行比较性研究,以确定二者的区别。方法通过显微镜观察精液液化过程中和粘稠度增高异常的精液的镜下特征,采用计算机辅助分析分别比较精液凝固状态下、液化状态下、正常精液和粘稠度增加状态下的精子运动特征。结果人精液液化过程中显微镜明显地观察到精浆中凝胶状物质的形态变化,而在粘稠度增加的精液中未能见到此类有形物质;配对资料分析显示,37例精液液化前后的精子运动状态存在显著性差异,精子在凝固的精浆中运动速度明显不同于高粘稠度精液中,而高粘稠度精液与正常精液的精子仅有精子的侧摆幅度有显著性差异,其它运动参数的差异无显著性。结论对精液中凝胶状物质的显微镜观察可以作为一种对精液液化进行客观判定的简易可行的手段,精液凝固和粘稠度增高对精子运动的影响是不同的。     

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.