Indirect fertility analysis in painters exposed to ethylene glycol ethers: sensitivity and specificity

Semen analysis has proven useful in the clinical diagnosis of infertility and is the most widely used method of monitoring the effects of occupational exposure on male fertility. Collection and analysis of semen samples in a field setting, however, require a highly motivated population and excellent technical resources, limiting the widespread application of the method. Techniques of monitoring male worker fertility using questionnaires to avoid some of the difficulties of semen analysis have been developed. These methods compare the rate of observed births for wives of workers with expected birth rates derived either from U.S. fertility tables or from unexposed workers. The present study compares the sensitivity of this questionnaire method with that of semen analysis in an evaluation of reproductive function in men exposed to ethylene glycol ethers. The reproductive function of 74 married painters exposed to ethylene glycol ethers was compared with that of 51 married controls employed at a shipyard. The groups differed in sperm count, but the questionnaire method showed no effect of exposure on fertility. This analysis suggests that the questionnaire assessment of fertility is less sensitive than semen analysis as a screening tool for male reproductive function.     

Exposure to hazardous substances and male reproductive health: a research framework

The discovery in the mid-1970s that occupational exposures to pesticides could diminish or destroy the fertility of workers sparked concern about the effects of hazardous substances on male reproductive health. More recently, there is evidence that sperm quantity and quality may have declined worldwide, that the incidence of testicular cancer has progressively increased in many countries, and that other disorders of the male reproductive tract such as hypospadias and cryptorchidism may have also increased. There is growing concern that occupational factors and environmental chemical exposures, including in utero and childhood exposures to compounds with estrogenic activity, may be correlated with these observed changes in male reproductive health and fertility. We review the evidence and methodologies that have contributed to our current understanding of environmental effects on male reproductive health and fertility and discuss the methodologic issues which confront investigators in this area. One of the greatest challenges confronting researchers in this area is assessing and comparing results from existing studies. We elaborate recommendations for future research. Researchers in the field of male reproductive health should continue working to prioritize hazardous substances; elucidate the magnitude of male reproductive health effects, particularly in the areas of testicular cancer, hypospadias, and cryptorchidism; develop biomarkers of exposure to reproductive toxins and of reproductive health effects for research and clinical use; foster collaborative interdisciplinary research; and recognize the importance of standardized laboratory methods and sample archiving.     

The healthy men study: an evaluation of exposure to disinfection by-products in tap water and sperm quality

BACKGROUND: Chlorination of drinking water generates disinfection by-products (DBPs), which have been shown to disrupt spermatogenesis in rodents at high doses, suggesting that DBPs could pose a reproductive risk to men. In this study we assessed DBP exposure and testicular toxicity, as evidenced by altered semen quality. METHODS: We conducted a cohort study to evaluate semen quality in men with well-characterized exposures to DBPs. Participants were 228 presumed fertile men with different DBP profiles. They completed a telephone interview about demographics, health history, water consumption, and other exposures and provided a semen sample. Semen outcomes included sperm concentration and morphology, as well as DNA integrity and chromatin maturity. Exposures to DBPs were evaluated by incorporating data on water consumption and bathing and showering with concentrations measured in tap water. We used multivariable linear regression to assess the relationship between exposure to DBPs and adverse sperm outcomes. RESULTS: The mean (median) sperm concentration and sperm count were 114.2 (90.5) million/mL and 362 (265) million, respectively. The mean (median) of the four trihalomethane species (THM4) exposure was 45.7 (65.3) microg/L, and the mean (median) of the nine haloacetic acid species (HAA9) exposure was 30.7 (44.2) microg/L. These sperm parameters were not associated with exposure to these classes of DBPs. For other sperm outcomes, we found no consistent pattern of increased abnormal semen quality with elevated exposure to trihalomethanes (THMs) or haloacetic acids (HAAs). The use of alternate methods for assessing exposure to DBPs and site-specific analyses did not change these results. CONCLUSIONS: The results of this study do not support an association between exposure to levels of DBPs near or below regulatory limits and adverse sperm outcomes in humans.     

Effect of occupational exposures on male fertility: literature review

The present review was aimed to determine the influence of working conditions, occupational exposures to potential chemical and physical reproductive toxic agents and psychological stress during work on male fertility. Significant associations were reported between impaired semen parameters and the following chemical exposures: metals (lead, mercury), pesticides (dibromochlorophane, 2,4-dichlorophenoxyacetic acid), ethylene glycol ethers and estrogens. The following physical exposures were shown to deteriorate sperm parameters: radiation (both ionized and microwaves) and heat. Psychological distress has another important contribution to infertility. Several studies indicated that stress has a negative impact on sperm parameters. Occupational parameters should be an important part of history taking among patients attending infertility clinics.     

Semen quality and reproductive sex hormone levels in Peruvian pesticide sprayers

To determine the association between occupational pesticide exposure and semen quality among sprayers using organophosphate (OPs) pesticides, 31 pesticide sprayers and 80 men who were not exposed were studied. Semen and blood samples were obtained one day after last pesticide application. Semen samples were analyzed for sperm concentration, percentage of sperm motility, percentage of normal morphology, and fructose and zinc concentrations. Blood samples were analyzed for sex hormones. Pesticide sprayers had significantly reduced age-adjusted seminal volume, percentage of motility, percentage of sperm with normal morphology, serum luteinizing hormone, serum testosterone levels, and seminal zinc concentration (a marker of prostate function), as well as significantly increased time of liquefaction, seminal pH, percentage of immature sperm morphology, and leukocyte concentration. These findings provide further evidence that occupational exposures to OP pesticides adversely affect semen quality and sex hormones.     

SEMEN ANALYSIS AT THE TURN OF THE CENTURY: AN EVALUATION OF POTENTIAL USES OF NEW SPERM FUNCTION ASSAYS

Semen analysis is a critical assay in the evaluation of infertility and may yield critical information regarding the etiology and prognosis of many types of reduced male fertility. However, basic semen analysis does not directly measure sperm fertilizing capacity, or many of the biochemical events both prior to and subsequent to fertilization. In the last two decades numerous assays of sperm function have been developed. These assays can be classified as: 1)Assays of general biochemistry and ultrastructure, 2) Assays of zona binding and oocyte penetration, and 3) Assays of postpenetration events. Sperm function assays not only allow an accurate diagnosis of many infertilities not diagnosed by the semen analysis, but can also lead to improved treatment modalities. In this review, basic semen analysis and many sperm function assays are briefly reviewed. Novel uses of sperm function are demonstrated in brief case studies.     

Residential distance to major roadways and semen quality,sperm DNA integrity,chromosomal disomy,and serum reproductive hormones among men attending a fertility clinic

Objective

We examined associations of residential distance to major roadways, as a proxy for traffic-related air pollution exposures, with sperm characteristics and male reproductive hormones.

Exposure to Polychlorinated Biphenyls (PCBs) and Male Reproduction

Polychlorinated biphenyls (PCBs) are a class of persistent organic pollutants that were widely used in the mid-20th century. Though their production and use was banned by most countries several decades ago, the general population continues to be exposed due to the persistence and bioaccumulation of PCBs. A number of human epidemiological studies have assessed the relationship between environmental PCB exposure and markers of male reproductive health, namely semen quality parameters (sperm concentration, motility, and morphology), sperm DNA integrity (DNA damage or chromatin fragmentation), and circulating reproductive hormone levels. Despite a wide range of study designs and locations, measurement methods, and PCB exposure levels, reports of inverse associations between PCBs and sperm motility have been consistent which may suggest a lack of exposure threshold for a PCB-related effect on sperm motility. Several studies have also reported inverse associations between PCBs and circulating testosterone levels in men, though the specific form of testosterone (i.e. total, bound, or free testosterone) associated with exposure has not been fully consistent between studies. In conclusion, although PCBs are no longer used and can be considered a legacy chemical, concerns regarding altered male fertility in relation to PCBs remain due to the existing human data demonstrating inverse associations with markers of male reproductive function coupled with recent evidence for continued population exposure.     

SPERM MORPHOLOGY AND CHROMATIN CONDENSATION BEFORE AND AFTER SEMEN PROCESSING

Semen analysis constitutes the most important investigation of male infertility. However, the true anomalies present in defective sperm cells have been only partially characterized. The integrity of the sperm chromatin may play the most important role, particularly in ICSI, where most of the natural selection mechanisms are bypassed. This study was carried out to characterize sperm morphology (strict criteria), to evaluate chromatin condensation and sperm count in native semen as well as after semen preparation by the swim-up technique, and to eventually evaluate any correlation between these parameters. Semen from 90 men was analyzed for the above parameters in both the fresh and processed semen. Whereas the sperm count decreased after sperm preparation by the swim-up technique in comparison to the value in the fresh semen (p<.001), there was an increase in the percentage of morphologically normal (p<.001) and chromatin-condensed sperm (p=.99). However, there was no correlation between sperm morphology, chromatin condensation, and sperm count either in the fresh or in the processed semen samples. These results suggest that sperm morphology, sperm count, and chromatin condensation are independent parameters that should be evaluated separately in the assessment of male fertility in an assisted reproduction program.     

Incidence and implications of altered semen quality on family planning

Alterations in the expression of the human genome, or interference with its products, can be induced in the male reproductive system by chemicals mimicking or antagonizing naturally occurring hormones. Opportunities exist for disruption at the hypothalamus, pituitary and testis levels. Recent concerns generated by the increased incidence of testicular cancer, congenital anomalies of the male genitalia and possible alterations in human semen quality have been linked to the environment.The report by Carlsen in 1992 [1] suggested that semen quality has deteriorated over the past six decades. More recent reports suggest that the decline may be globally non-uniform and regional in nature. The effects of any such declines upon overall pregnancy rates are generally unknown, although some studies have attempted to address them. A preliminary review of the impact of a small decrease in sperm concentrations suggests that a directly measurable reduction in fecundity does not occur, but that future problems could be anticipated. Decrements in semen quality will alter the epidemiological probabilities of pregnancy due to coitus on different cycle days and may thereby change the duration of the fertile time. Current understanding of the implications of altered semen quality on relative fertility is not sufficient to change our current teaching and practice of NFP.     

The effects of inbreeding on testicular sperm concentration in Peromyscus polionotus

Inbreeding effects on fitness have most often been quantified via juvenile mortality. However, inbred adults may suffer from inbreeding depression if their fertility or fecundity is compromised as a consequence of inbreeding. Here, the effects of inbreeding on male fertility in oldfield mice, Peromyscus polionotus, were examined. Testicular sperm concentration was assessed in 93 males, 68 of which were paired for breeding. Forty of the 68 paired males failed to produce offspring. Total testicular sperm count, sperm count (g testis)(-1), and testis mass all declined significantly with increasing inbreeding coefficient. Sperm concentration did not significantly impact reproductive success. Although sperm concentration in males of most species can decline to low levels before reproductive impairment is detectable, the declines in testicular sperm concentration found here suggest that inbreeding can affect fertility in adult males. Furthermore, monitoring testicular sperm concentration could provide a mechanism to monitor potential declines in reproductive performance before population-level reproductive success is irreparably impaired. The implications for the management of small, captive and wild populations may be substantial.     

Segregation of sperm subpopulations in normozoospermic infertile men

Sperm function is essential for fertilization and embryogenesis yet semen contain a heterogeneous population of sperm. This study was designed to evaluate two different sperm populations separated by the density gradient method. Semen from 25 idiopathic normozoospermic infertile men was processed by double density gradient centrifugation and evaluated for sperm present in the 50% (upper) layer and the 90% (lower) layer for reactive oxygen species (ROS), sperm chromatin integrity, and morphology. The population of sperm in the 90% layer showed significantly lower ROS levels (22.90 (0.92, 85.32) vs. 382.03 (158.30, 1409.51) and lower DNA fragmentation index (DFI) (24.26 (22.54, 25.50) vs. 29.93 (28.48, 31.25) and higher number of sperm with normal morphology (55 (45.0, 60.0) vs. 32.5 (20, 40) compared to sperm in the 50% layer. However, in the original raw semen, sperm DFI (27.02 (26.19, 27.76)) and percentage high DNA stainability (% HDS) (3.1 (2.40, 3.78)) cells were significantly higher compared to the 90% layer population. Density gradient separation of the sperm subpopulation from the original semen favors the selection of sperm with genome integrity, low levels of ROS, and normal morphology. Therefore presence of pathological sperm in the semen may disrupt the function of normal spermatozoa, and hence the selection of the normal sperm subpopulation may be a better candidate for assisted conception. Further studies are required to evaluate the gradient separated sperm population in assisted reproductive techniques (ART).     

Hair mercury (Hg) levels,fish consumption and semen parameters among men attending a fertility center

General population exposure to methylmercury (MeHg), the most common organic mercury compound found in the environment, occurs primarily through the consumption of contaminated fish and shellfish. Due to limited studies and lack of consideration of effect modification by fish consumption, it remains uncertain if exposure to mercury affects semen parameters. Thus, we investigated whether hair Hg levels, a biomarker of mercury exposure, were associated with semen parameters among men attending an academic fertility center, and whether this relationship was modified by intake of fish.This analysis included 129 men contributing 243 semen samples who were enrolled in the Environment and Reproductive Health (EARTH) Study between 2005 and 2013, and had data of hair Hg, intake of fish and semen parameters available. Hair Hg levels were assessed using a direct mercury analyzer. Intake of fish was collected using a validated food frequency questionnaire. Semen parameters were analyzed following WHO 2010 evaluation criteria. Generalized linear mixed models with random intercepts accounting for within-man correlations across semen samples were used to evaluate the association of hair Hg levels and semen parameters adjusting for age, BMI, smoking status, abstinence time and alcohol intake. Effect modification by total fish intake (≤1.68 vs.?>1.68 servings/week) was tested.The median hair Hg levels of the men was 0.72 ppm and ranged from 0.03 to 8.01 ppm; almost 30% of the men had hair Hg levels >1 ppm. Hair Hg levels were positively related with sperm concentration, total sperm count, and progressive motility, after adjusting for potential confounders and became attenuated after further adjustment for fish intake. Specifically, men in the highest quartile of hair mercury levels had 50%, 46% and 31% higher sperm concentration, total sperm count and progressive motility, respectively, compared to men in the lowest quartile. These associations were stronger among men whose fish intake was above the study population median. Semen volume and normal morphology were unrelated to hair Hg levels.These results confirmed exposure to MeHg through fish intake and showed the important role of diet when exploring the associations between heavy metals and semen parameters among men of couples seeking fertility care. Further research is needed to clarify the complex relationship between fish intake and Hg, and potential effects on male reproductive health, specifically, semen parameters.     

The relationship between human semen parameters and environmental exposure to polychlorinated biphenyls and p,p'-DDE

Scientific and public concern exists about potential reproductive health effects of persistent chlorinated organic chemicals, such as polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDT), and dichlorodiphenyldichloroethylene (DDE, the most stable daughter compound of DDT). To explore the hypothesis that environmental exposures to PCBs and DDE are associated with altered semen parameters, we conducted a cross-sectional study of 212 male partners of subfertile couples who presented to the Massachusetts General Hospital Andrology Laboratory. Semen parameters were analyzed as both a continuous measure and dichotomized based on World Health Organization reference values for sperm concentration (< 20 million/mL), motility (< 50% motile), and Kruger strict criteria for morphology (< 4% normal). The comparison group for the dichotomized analysis was men with all three semen parameters above the reference values. In serum, 57 PCB congeners and p,p -DDE were measured by congener-specific analysis using gas chromatography with electron capture detection. There were dose-response relationships among PCB-138 and sperm motility (odds ratio per tertile, adjusted for age, abstinence, and smoking, and p-value for trend were, respectively, 1.00, 1.68, 2.35, and p-value = 0.03) and morphology (1.00, 1.36, 2.53, p-value = 0.04). There was limited evidence of an inverse relationship between sum of PCBs, as well as those PCBs classified as cytochrome P450 enzyme inducers, with sperm motility and sperm morphology, as well as limited evidence of an inverse association between p,p -DDE and sperm motility. The lack of a consistent relationship among semen parameters and other individual PCB congeners and groupings of congeners may indicate a difference in spermatotoxicity between congeners.     

Endocrine disruptors and human health--is there a problem? An update.

It has been hypothesized that environmental exposure to synthetic estrogenic chemicals and related endocrine-active compounds may be responsible for a global decrease in sperm counts, decreased male reproductive capacity, and breast cancer in women. Results of recent studies show that there are large demographic variations in sperm counts within countries or regions, and analyses of North American data show that sperm counts have not decreased over the last 60 years. Analyses of records for hypospadias and cryptorchidism also show demographic differences in these disorders before 1985; however, since 1985 rates of hypospadias have not changed and cryptorchidism has actually declined. Temporal changes in sex ratios and fertility are minimal, whereas testicular cancer is increasing in most countries; however, in Scandinavia, the difference between high (Denmark) and low (Finland) incidence areas are not well understood and are unlikely to be correlated with differences in exposure to synthetic industrial chemicals. Results from studies on organochlorine contaminants (DDE/PCB) show that levels were not significantly different in breast cancer patients versus controls. Thus, many of the male and female reproductive tract problems linked to the endocrine-disruptor hypothesis have not increased and are not correlated with synthetic industrial contaminants. This does not exclude an endocrine-etiology for some adverse environmental effects or human problems associated with high exposures to some chemicals.     

Association between a biomarker of exposure to polycyclic aromatic hydrocarbons and semen quality

Objectives

Growing evidence supports the reproductive and developmental toxicity of polycyclic aromatic hydrocarbons (PAHs) from prenatal and postnatal exposure, but the results of epidemiological studies regarding harmful effects of PAHs exposure on male reproductive system still remain limited and inconclusive. The aim of the present study was to investigate the relationship between 1-hydroxypyrene, a biomarker of polycyclic aromatic hydrocarbons exposure and semen quality.

Materials and Methods

The study population consisted of 277 men attending an infertility clinic for diagnostic purposes and having normal semen concentration of 20–300 mln/ml or slight oligozoospermia (semen concentration: 15–20 mln/ml) (WHO 1999). All the men were healthy and under 45 years of age. All participants were interviewed and provided a semen sample. The interview included questions concerning demographics, socio-economic status, medical history related to past diseases which may have an impact on semen quality, lifestyle factors and occupational information. Concentrations of 1-hydroxypyrene (1-OHP) in the urine samples were analyzed using high performance liquid chromatography (HPLC).

Results

A positive association was found between the level of 1-OHP in urine and sperm neck abnormalities as well as the percentage of static sperm cells (p = 0.001, p = 0.018, respectively). Additionally, exposure to PAHs measured by 1-OHP in urine decreased semen volume and the percentage of motile sperm cells (p = 0.014, p = 0.0001, respectively).

Conclusions

Presented findings indicate that the environmental level of PAHs exposure adversely affects male semen quality. The future large-scale studies should incorporate different biomarkers to generate a more accurate and full assessment of the effects of PAHs exposure on male fertility.     

Environmental fertility research at the turn of the century

Research into environmental causes of impaired human fertility is still immature. Knowledge on the strengths and limitations of several research options, the main sources of bias, the occurrence of infertility and subfecundity in several countries, the patterns of contraceptive behavior, and the distribution of sperm counts at the population level has advanced, but the much-debated questions about temporal and regional trends in fertility and semen quality remain unsettled. Refined biological markers of male fecundity and markers feasible for large-scale studies of female fecundity are needed. The effects of several life-style factors have been characterized successfully, but the detrimental effects of environmental exposures have not. Most studies are either inconclusive or fail to demonstrate any important risks. If comprehensive knowledge on biological mechanisms is a criterion of maturity, epidemiologic and toxicologic fertility research is young. The new millennium will be challenged to determine the role played by the environment in developed countries, wrong exposure time windows, too crude or inappropriate measures of fertility, and overlooked critical exposures.     

Flow cytometry applications in the evaluation of sperm quality: semen analysis, sperm function and DNA integrity

Flow cytometry (FCM) has been extensively used to study mammalian sperm in the areas of reproductive toxicology (to monitor effects from environmental, occupational and therapeutic exposures), veterinary science (to preselect the gender of offspring by sorting X- and Y-chromosome-bearing sperm) and clinical andrology (to assess individual fertility potential). Using FCM, a variety of sperm features can now be rapidly measured on a cell-by-cell basis such as sperm count, viability, acrosomal integrity, mitochondrial function and DNA integrity; the last one is involved in postfertilization failure and embryo toxicity. It is foreseen that only a multiplex approach, which includes FCM assays together with the new genomics/proteomics methods, could increase the predictive power of fertility status and help identify susceptible subpopulations of men at risk for infertility, spontaneous abortions and birth defects.     

输精管吻合术后精子超微结构观察

目的:观察输精管吻合术后精子超微结构的变化。方法:采用光学显微镜、扫描和透射电子显微镜对20例接受输精管吻合术者和10例正常生育男性精子的形态和超微结构改变进行观察、统计、分析。结果:①光镜和扫描电镜分析显示吻合术后精液中正常形态精子显著减低;②两组精子尾畸在光镜下有明显差异,在扫描电镜下无差异;③两组精子头畸在扫描电镜下有显著性差异;④在透射电镜下吻合术后精子多数均属于复合型精子结构异常,大部分均有多个部位和多个细胞器结构异常。结论:吻合术后精子有多种类型和复合型的异常改变,这些改变与不育男子精子的改变相似。提示:输精管结扎术后精子形态学的改变,是使吻合术后生育能力降低的主要原因之一。