Relation between morphologically abnormal spermatozoa and pregnancies obtained during a twenty-year follow-up period

The clinical fertility of 1077 men investigated with semen analysis including detection of morphologically abnormal spermatozoa during the years 1950-52 was studied 20 years later using a questionnaire replied to by 785 (72.9%), There was a significant correlation ( P <0.01) between increasing number of abnormal spermatozoa and decreasing chance of getting living children, but no relation to abortions and pathological pregnancies. Moreover, there was a significant correlation P <0.01) to time-interval to first pregnancy. Median values for those obtaining living children was 25% abnormal spermatozoa compared with 28% for those not doing so. Up to 60% abnormal spermatozoa, fertility was not reduced, and the borderline between normal and reduced male fertility is therefore defined to be 60% morphologically abnormal spermatozoa.     

Subnormal sperm parameters in conventional semen analysis are associated with discrepancies between fertilization and pregnancy rates in in-vitro fertilization and embryo transfer

Three hundred and twenty-eight consecutive treatment cycles in 168 couples were analysed retrospectively in order to examine the influence of conventional semen analysis results on the outcome of in-vitro fertilization and embryo transfer with respect to the occurrence of both fertilizations and pregnancies. All treatments were performed under maximally standardized and controlled conditions. Each of the three main determinants of the spermiogram, namely the concentration, motility and morphology of sperm in seminal plasma, was of significant importance for fertilization and subsequent pregnancy. Best correlations were achieved by counting the number of progressively (a+b) motile sperm and the number of normally formed sperm in seminal plasma. The pregnancy rate was reduced significantly in cases in which the sperm concentration was < 10 x 10(6) ml-1 (P < 0.01), or in which there was < 40% progressively motile sperm (P < 0.001), or < 30% normally formed sperm (P < 0.001). If more than one parameter in the spermiogram was abnormal, the fertilization rate depended mainly on the most disturbed sperm parameter. The implantation rate as well as the pregnancy rate was reduced significantly in patients with low progressive sperm motility and normal morphology rates. The difference could only be attributed partially to the lower number of embryos replaced. In conclusion, subnormal sperm quality seems to interfere with developmental stages beyond the process of fertilization.     

1985～2008年间我国正常男性精液质量变化分析

目的:研究1985～2008年间国内正常男性精液质量变化趋势。方法:检索历年发表的精子质量方面的文献,系统收集相关资料和数据,利用散点图和直线回归法,分析近24年正常男性精液精子密度、精液量及精子总数的变化。结果:1985～2008年间国内正常男性精液质量参数中精液量没有明显变化(P>0.05)。1985～1994年精子密度和精子总数不存在与时间相关的变化(P>0.05),但自1995年后的14年以来,精子密度和精子总数呈现显著下降趋势(P<0.05),精子密度从1995年81.5×106/ml下降到2008年66.7×106/ml,每年降低1.40%;精子总数从1995年257.2×106下降到2008年的185.9×106,每年下降2.15%。结论:1985～1995年间国内正常男性精液质量未见明显变化,而1995～2008年14年间精子密度和精子总数明显降低,很可能与进入重化工时代引起的日益加重的环境污染有关。     

精液白细胞含量变化对精子顶体酶活性的影响

目的观察精液白细胞含量变化对精子顶体酶活性的影响。方法在抗炎治疗前后采用正甲苯胺蓝过氧化物酶法对精液中白细胞进行定量测定,并与精子顶体酶活性进行相关性分析。结果383例男性不育就诊者中有106例精液白细胞＞1×10~6个/ml,其精子项体酶活性(12.81±9.23)μIU/10~6精子；经抗炎治疗后75例精液白细胞≤1×10~6个/ml,设为有效组,其精子顶体酶活性(20.65±10.05)μIU/10~6精子,余31例精液白细胞＞1×10~6个/ml,设为无效组,其精子顶体酶活性(13.26±9.81)μIU/10~6精子。治疗后两组精子顶体酶活性有显著性差异(P＜0.01)。结论精液白细胞含量变化可影响精子顶体酶活性,精液白细胞症经抗炎治疗改善后可提高精子顶体酶活性。     

The influence of processed total non-forward and non-motile sperm count on the outcome of artificial insemination with the husband’s semen

BackgroundArtificial insemination with the husband’s semen (AIH) is an economical and noninvasive method of infertility treatment. However, AIH’s pregnancy rate is much lower than in vitro fertilization (IVF) as its multiple and complex uncertainty factors. Semen quality has been one of the main factors which affect the pregnancy outcome of AIH.MethodsThe relevant parameters of 1,142 AIH cycles were retrospectively studied, including the general parameters and the semen quality parameters among clinical pregnancy, biochemical pregnancy, non-pregnancy group, age, infertility duration, infertility type, body mass index (BMI), cycle count, morphology in previously semen examination, and semen quality parameters on the day of AIH.ResultsThe statistically significant difference was only found on processed total non-forward and non-motile sperm count (N-TFMSC). The mean processed N-TFMSC in the biochemical pregnancy group was 6.37±4.27 million, significantly higher than the other two groups (vs. 4.40±3.15 million or vs. 4.48±3.60 million, P<0.05). The study was then divided into two groups according to processed N-TFMSC, Group 1 ≤5.0 million, and Group 2 >5.0 million. A statistical increase in biochemical pregnancy rate was observed when the processed N-TFMSC was >5.0 million (2.72% vs. 0.90%).ConclusionsProcessed N-TFMSC may be one of the independent factors on AIH’s outcome; it should be given equal attention the same as processed total forward motile sperm count (TFMSC).     

Single semen analysis as a predictor of semen quality： clinical and epidemiological implications

It is generally thought that a single ejaculate is a bad predictor of semen quality of a subject, because of significant intra-individual variation. Therefore, we investigated the degree to which the results of a first semen analysis differ from that of a second analysis among men from a how the two different semen results mirrored the overall general population in Norway. In addition, we analysed semen quality assessment. A total of 199 volunteers participated in the study and delivered two semen samples with an interval of 6 months. The semen parameters were determined according to the World Health Organization （WHO） 1999 guidelines, which were also used to determine whether semen quality was normal or abnormal. In addition, the DNA fragmentation index （DFI） was determined using the Sperm Chromatin Structure Assay. The two samples from each individual were very similar with regard to standard semen parameters and DFI （rs： 0.67-0.72）, and there were no significant systematic differences between the two samples. The result of the first sample （normal/abnormal） was highly predictive of the overall conclusion based on the two samples （sperm concentration： in 93% of the cases （95% confidence interval [CI]： 89%-96%）; sperm motility： in 85% of the cases （95% CI： 79%-89%）; overall semen quality： in 85% of the cases （95% CI： 80%-90%）. In epidemiological studies, one ejaculate is a sufficient indicator of semen quality in a group of subjects. In a clinical situation, when the question is whether the semen quality is normal or not, the first ejaculate will, in at least 85% of cases, give a correct overall conclusion.     

Evaluation of donor semen quality provided by six sperm banks: a retrospective study of 1877 artificial insemination cycles

This study aimed at evaluating the impacts of sperm quality of six national sperm banks on pregnancy rates (PRs) of artificial insemination with donor sperm (AID) in China. A large retrospective analysis was performed on 1877 insemination cycles in 1209 women in a unique setting during a 3.5-year period. Global PRs of 22.1% per cycle and 34.2% per patient were achieved. The PRs of the six banks varied from 15.5% to 29.0% (P = 0.011). Significant differences were observed in the quality of donor semen provided by the six sperm banks. Moreover, in some banks, the poor sperm quality was related to the suboptimal PRs. However, in certain banks, high values of sperm parameters did not result in satisfactory PRs accordingly. These data demonstrated that variability of donor semen quality existed in the different banks. But, sperm parameters after thawing may not be detrimental factors affecting the success rate of AID treatment. Further studies are needed to seek potential molecular markers for predicting fertility potency of donor sperm.     

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.     

Comparative study of the effects of three semen preparation media on semen analysis,DNA damage and protamine deficiency,and the correlation between DNA integrity and sperm parameters

Semen samples collected from 28 male partners of infertile couples were divided into three equal aliquots and prepared with three selected media,such as PureSperm （Nidacon,Gothenburg,Sweden）,Sil-Select Plus^TM （Fertipro,Beemem,Belgium） and SpermGrad^TM（Vitrolife,Gothenburg,Sweden）. The differences in mean percentages of semen parameters were assessed by repeated measures analysis. Correlations of sperm DNA damage,as measured by terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL） assay,and of protamine deficiency,as measured by chromomycin A3 （CMA3） staining with sperm parameters,were determined by Pearson＇s correlation. After preparation with all three media,sperm concentrations decreased （P〈0.05） while percentages of sperm with normal morphology increased （P〈0.05）. Percentages of sperm motility,rapid motility and progressive motile concentration （PMC） increased （P〈0.05） for each ofthese parameters,PureSperm preparation gave the best results （P〈0.05）. The percentage of DNA damage decreased in the PureSperm and Sil-Select Plus preparations （17.9% and 31.3%,respectively,P〈0.05） and increased in the SpermGrad preparation （56.3%,P〈0.05）. Protamine deficiency also decreased in all three kinds of media,59.3%,47.7% and 40.3% for PureSperm,Sil-Select Plus and SpermGrad preparations,respectively （P〈0.05）. The percentage of DNA-damaged sperm was negatively correlated with the percentages of sperm motility,rapid motility and PMC,but was positively correlated with static motility （P〈0.05）. This comparative study and correlation analysis revealed that PureSperm preparation yielded sperm with the best motility and the lowest percentage of protamine deficiency. The Sil-Select Plus preparation yielded sperm with the lowest amount of DNA damage. The SpermGrad preparation had a high percentage of sperm with normal morphology,but also had the highest percentage of sperm with DNA damage. Sperm DNA damage was correlated with percentages of sperm motility,rapid motility,static motility and PMC.     

A randomised trial comparing conventional semen parameters,sperm DNA fragmentation levels and satisfaction levels between semen collection at home and at the clinic

The aim of the randomised trial was to compare conventional semen parameters, sperm DNA fragmentation levels and satisfaction levels between semen samples collected at home and at the clinic. We recruited 110 men with a history of infertility for at least 1 year from the outpatient andrology clinic. Each man collected two semen samples, one at home and one at the clinic. Men were randomly assigned into the home first (n = 55) or clinic first (n = 55) groups. The primary outcome was sperm concentration. There was no significant difference in sperm concentration, sperm DNA fragmentation levels or other conventional semen parameters between home first and clinic first samples (p > .05), while satisfaction levels were significantly higher for home first samples (p < .01). Consistent results were obtained when comparing home-collected and clinic-collected samples within individuals. Men can be offered the option to collect semen samples at home for examination or assisted reproduction without compromising semen quality, especially for those with difficulty in producing semen samples at the clinic.     

Relationship between human sperm lipid peroxidation, comprehensive quality parameters and IVF outcome

The membranes of human spermatozoa contain an extremely high concentration of polyunsaturated fatty acids and are therefore susceptible to lipid peroxidation damage. The aim of this study was to retrospectively determine the association between the lipid peroxidation levels of washed spermatozoa, as indicated by thiobarbituric-acid-reactive substance concentration, and: (a) semen quality evaluated by basic routine, biochemical, cytological and quantitative ultramorphological analyses; (b) IVF fertilization rate. Semen samples from 45 male partners of couples who had been referred for IVF treatment due to a female infertility factor were evaluated for quality as well as for thiobarbituric-acid-reactive substance concentrations. The latter were found to have a negative correlation with total sperm count, semen volume, zinc/fructose ratio, and the integrity of sperm acrosome and axonema. It was suggested that lipid peroxidation has a deleterious effect on the ultramorphological status of the sperm cells and, thereby, on the male fertilization potential. The content of the seminal fluid, about 30% of which is produced by the prostate, may protect spermatozoa from this destructive process. A negative correlation was also found between thiobarbituric-acid-reactive substance concentrations and IVF fertilization rate. When the patients were subdivided into fertilizing (fertilization rate > 0%) and nonfertilizing (fertilization rate = 0%) subgroups (n = 33 and n = 12, respectively), the former exhibited significantly lower thiobarbituric-acid-reactive substance concentrations than the latter. A new IVF fertilization index based on the lipid peroxidation level was established. This index had a predictive power of 93% (94% sensitivity and 92% specificity). The clinical value of this index should be further verified.     

Reassessment of sperm morphology of archival semen smears from the period 1980--1994

Several reports have suggested that sperm counts of normal men have declined in many geographical regions during the last decades. Deterioration of sperm morphology has also been reported in some studies covering long sample collecting periods. The original semen analysis data of our semen laboratory from the period 1980--1994 showed a significant decline in the proportion of spermatozoa with normal morphology. The finding was, however, questioned because of changes in sperm morphology assessment criteria during the study period. In the present study 1745 smears were re-analysed to cover evenly the whole study period. The samples were examined in random order by using strict assessment criteria. Multiple linear regression analysis of the re-analysed data showed no effect of the year of sample delivery on sperm morphology between the years 1980 and 1994. However, there was a significant decline in the proportion of normal spermatozoa with later year of men's birth.     

Geographical differences in semen characteristics: Comparing semen parameters of infertile men of the United States and Iraq

The declining trend of male fecundity is a major global health and social concern. Among numerous other confounding factors, variations in male fertility parameters in different regions have repeatedly been suggested to be influenced by geographic locations. The impact of overall lifestyle, behavioural patterns, ethnicity, work stress and associated factors upon health differ greatly between developed and developing countries. These factors, individually or in combination, affect male reproductive functions ensuing the discrepancies in semen qualities in connection with geographic variations. However, reports comparing semen characteristics between developed and developing countries are sparse. The present study finds its novelty in presenting a comparison in semen parameters of infertile men in the United States (n = 76) that fairly represents the population of a highly developed region and Iraq (n = 102), the representative of male populations of a developing region. Samples were collected and analysed according to WHO (WHO laboratory manual for the examination and processing of human semen, WHO; 2010) criteria by means of the Mann–Whitney test. The US population demonstrated lower sperm concentration, total count, and total and progressive sperm motility with a higher seminal total antioxidant capacity (TAC) as compared to the Iraqi population. This report encourages further investigations concerning the confounding factors leading to such alterations in semen qualities between these two geographic areas.     

Evaluation of trend in semen analysis for 11 years in subjects attending a fertility clinic in India

Aim: The data on semen analysis of subjects attending the Fertility Clinic at NIHFW (National Institute of Health and Family Welfare) Munirka, New Delhi for the last 11 years were analyzed to verify the claims and speculations on declining sperm counts in men. Methods: Approximately 10 % of the records every year starting from 1990 to 2000 (numbering 1176 in total) were randomly selected for analysis. Subjects with azoospermia or severe oligozoospermia were excluded from analysis. Results: The average age of the men attending the infertility clinic was 31.2 years. The average semen volume and sperm count were found to be (2.6 ±0.1) mL and (60.6 ± 0.9) × 106/mL, respectively. No significant decline in sperm counts was observed in any year during the entire study period. Only 1.8 % of the total number of sperm counts in the random sampling were less then 20 × 106/mL. On the basis of WHO criteria on motility, the total percentage of non-progressive and non-motile sperm in the ejaculate was higher (63 %)     

精液分析中精子浓度室内质量控制方法的研究

目的:建立一种可行的精子浓度检测的室内质控方法。方法:设定20×106和80×106两种浓度的冷冻精液作为高、低值室内质控品(IQC),QC-BEADSTM质控珠(以下简称质控珠)作为对照。采用双盲法,4名技术员分别通过计算机辅助精液分析(CASA)对上述假定的室内质控品及质控珠进行精子浓度的测定,绘制出各自的质控图(Xbar图和Sbar图),连续检测1个月,计算出相应的高、低浓度值批内及批间变异系数(CV%),并进行比较。结果:①假定室内质控品高、低值批内变异系数分别CV3.5%,CV2.4%;批间分别为CV10.2%、CV9.6%。质控珠高、低值批内分别为CV5.1%,CV7.1%,批间分别CV7.1%,CV8.2%。两组高、低值的批内变异均<10%,批间变异均<15%,符合(Levey-Jennings)L-J质量控制原则,均能达到室内质控目的。②两组间高、低值的批内变异系数及批间变异系数无显著性差异(P>0.05),提示假定质控品可代替质控珠行实验室内部质量控制。结论:我们建立的精子浓度测定的室内质控方法是简单、易行、可靠。     

Elimination of bacteria from human semen during sperm preparation using density gradient centrifugation with a novel tube insert

The occurrence of bacteria in sperm samples intended for in vitro fertilisation can compromise the outcome of assisted reproductive techniques. Effective semen processing procedures should therefore be implemented to remove bacteria from semen. Unfortunately, technique failure does occur whereby bacteria can be found in processed sperm preparations. To improve the effectiveness of semen processing, a novel centrifuge tube insert was developed to facilitate the layering of density gradients and semen, and to prohibit the re‐infection of purified sperm pellets. The purpose of this study was to: (i) determine the prevalence and type of bacteria present in semen of patients participating in the Unit’s assisted reproduction program and (ii) evaluate the effectiveness of density gradient centrifugation with the novel tube insert, for the elimination of bacteria and yeast from spiked human semen samples. A survey in 2007–2010 indicated that 50% of semen samples were found to have positive bacterial cultures. Semen processing by means of density gradient centrifugation with the novel tube insert eliminated significantly more in vitro derived (spiked) bacteria and yeast from semen compared to processing without the insert (P < 0.004). Therefore, it is highly recommended that the centrifuge tube insert, ProInsert?, be incorporated into assisted reproductive programs.     

Reassessment of the accuracy of traditional sperm characteristics and adenosine triphosphate (ATP) in estimating the fertilizing potential of human semen in vivo

Receiver operating characteristic curves and accuracy parameters were computed for traditional sperm characteristics (concentration, motility, morphology) and the number of peroxidase negative cells, and the concentration of adenosine triphosphate (ATP) in semen from populations of fertile and infertile men, and men who achieved a pregnancy after varicocele treatment. The percentage and concentration per millilitre of spermatozoa with rapid linear progressive motility, and the ATP concentration, provided the best discrimination between fertile and treated fertile from infertile men. The misclassification rate was higher for sperm morphology, total progressive motility and viability, whereas sperm concentration and the total sperm count per ejaculate had the worst discriminating power. The number of peroxidase negative cells per 100 spermatozoa was highly specific in identifying men who achieved pregnancy after varicocele treatment. The lower limit of normality of sperm characteristics was remarkably different between fertile men and men achieving pregnancy after treatment or during infertility work-up.     

The outcome of clinical pregnancies following intracytoplasmic sperm injection is not affected by semen quality

Summary.  The objective of this study was to investigate the impact of severe oligoasthenoteratozoospermia (OAT) on pregnancy outcome. For this purpose 279 consecutive intracytoplasmic sperm injection (ICSI) cycles were retrospectively evaluated and compared to 436 consecutive IVF cycles performed during the same time frame. Group A ( n = 62) included ICSI patients with severe OAT; group B ( n = 217) included patients who underwent ICSI for other indications; and group C ( n = 436) included couples who underwent standard IVF. The mean age of female patients and mean number of embryos transferred were comparable in all groups. No difference was observed regarding implantation, clinical pregnancy, delivery and miscarriage rates between all three groups, but fertilization rate was significantly lower in group A than in groups B and C. It is concluded that couples undergoing ICSI with severe male infertility (OAT) have a slightly reduced fertilization rate but their chances of delivery and pregnancy loss are similar to those of other patients undergoing clinical ICSI and IVF with non-male infertility.     

Effects of leptin on sperm count and morphology in Sprague‐Dawley rats and their reversibility following a 6‐week recovery period

Altered epididymal sperm count and morphology following leptin treatment has been reported recently. This study examined the effects of 42 days of leptin treatment on sperm count and morphology and their reversibility during a subsequent 56‐day recovery period. Twelve‐week‐old male Sprague‐Dawley rats were randomised into four leptin and four saline‐treated control groups (n = 6). Intraperitoneal injections of leptin were given daily (60 μg Kg^?1 body weight) for 42 days. Controls received 0.1 ml of 0.9% saline. Leptin‐treated animals and their respective age‐matched controls were euthanised on either day 1, 21, 42 or 56 of recovery for collection of epididymal spermatozoa. Sperm concentration was determined using a Makler counting chamber. Spermatozoa were analysed for 8‐hydroxy‐2‐deoxyguanosine and DNA fragmentation (Comet assay). Data were analysed using anova . Sperm concentration was significantly lower but fraction of abnormal spermatozoa, and levels of 8‐hydroxy‐2‐deoxyguanosine were significantly higher in leptin‐treated rats on day 1 of recovery. Comet assays revealed significant DNA fragmentation in leptin‐treated rats. These differences were reduced by day 56 of recovery. It appears that 42 days of leptin treatment to Sprague‐Dawley rats has significant adverse effects on sperm count and morphology that reverse following discontinuation of leptin treatment.