A survey on the status of semen analysis in 118 laboratories in China

Collecting baseline information on how laboratories perform testing is a reasonable first step towards establishing intra- and inter-laboratory standardization and quality control for semen analysis. We carried out a survey of the laboratories performing the testing in China's Mainland. A questionnaire, composed of 36 questions covering all aspects of semen analysis, was designed, and a copy was distributed to each of the 145 laboratories. Of these, 118 laboratories completed the questionnaires. The survey results showed that semen volume was measured visually in 53.6% （59/110） of the responding laboratories, and 70.9% （73/103） of laboratories analysed incompletely liquefied semen without any treatment. In addition, both manual-microscopic and computer-assisted semen-analysis systems were applied to analyse sperm concentration, motility and morphology. However, more than five methods were employed in routine sperm staining. An enzyme-linked immunosorbent assay was commonly used for determining whether antisperm antibodies were present. Several seminal biochemical markers were analysed in only 27.1% （32/118） of the responding laboratories. Generally, there was a lack of intra- and inter-laboratory quality control measures for semen analysis in all laboratories responding to this survey. In conclusion, the methods of semen analysis and the interpretation of test results in the surveyed laboratories differed markedly. In particular, many laboratories employed methods other than those recommended by the World Health Organization Laboratory Manual for the Examination of Human Semen and Sperm- cervical Mucus Interaction （1999）. These findings suggest an urgent need for the standardization of semen analysis with acceptable quality controls for each parameter to make the results repeatable and meaningful.

Semen quality analysis and the idea of normal fertility

The World Health Organization （WHO） has extensively revised its manual for semen analysis, and seeks to provide reference limits for semen quality parameters. This raises the question of what is meant by ＇normal＇ （excluding the use of this term to denote a Gaussian distribution）. It could be taken in a purely statistical sense, using a biologically arbitrary cut-off point to denote an abnormal level, typically the extreme 5 percent of the population. Alternatively, ＇normal＇ could be defined according to the biological concept of normality and abnormality, in terms of the point at which biological function becomes impaired. Either of these can be used in descriptive epidemiology, for example, to study trends, but in the case of fertility, both semen quality and functional fertility （time to pregnancy） are continuous variables with no clear threshold. The WHO manual uses the biological meaning of normal, in that it provides the semen parameter distributions for men who have recently fathered pregnancies that took 12 months or less to conceive. However, what is really needed is the same information the other way around： given a particular semen test result, what should be expected in terms of ability to conceive, and how long it is likely to take. In considering epidemiological research, the focus has been mainly on internal comparisons, rather than reference limits, but it would be beneficial if more attention were paid to the absolute levels and to what these mean in terms of function--in other words, if the data were better calibrated biologically.

Semen analysis in 21st century medicine： the need for sperm function testing

Sperm function testing, once commonly performed for the infertile couple before employing assisted reproductive technology （ART）, has fallen out of favour in many reproductive medicine centers throughout the world. Indeed, the most recent addition of the ＇World Health Organisation （WHO） Laboratory Manual for the Examination and Processing of Hu- man Semen＇ now groups many of these procedures into a section termed Research Procedures. In large part, this reflects the current clinical practice of bypassing the in-depth evaluation of the male partner, while assuming that if a spermatozoon can be found for intracytoplasmic sperm injection （ICSI）, it must be a healthy cell capable of achieving fertilization. Never- theless, sperm function testing can provide valuable clinical insights into defects causing male infertility. Admittedly, in some cases, functional sperm deficiencies can be overcome using an ART. In other cases, couples will be empowered by the knowledge of the cause of their infertility, and for some couples, perhaps even the likelihood of ICSI success （relative to the spermatozoa）. The knowledge allows them to make truly informed reproductive decisions, including （perhaps） the de- cision to seek donor insemination, to adopt or to remain childless. Knowledge of the cause of their infertility may provide closure for couples and a sense of confidence regarding their choice of reproductive treatment.

Semen analysis： a new manual and its application to the understanding of semen and its pathology

This article reviews the latest edition of the World Health Organization＇s manual on seme instructional guide. The methodology used in the assessment of the usual variables in semen many of the less common, but very valuable, sperm function tests. Seminal fluid preparatio such as in vitro fertilization and intrauterine insemination are also outlined in the manual. useful techniques for the assessment of seminal fluid. It will be a very useful manual for an analyses of seminal fluid.

Semen analysis from an epidemiologic perspective

The fifth edition of the World Health Organization （WHO） manual for semen analysis includes for the first time reference values for human semen characteristics. This paper considers whether such values will help to resolve the intensely debated data indicating temporal and geographical shifts in sperm counts and hypotheses that anthropogenic activities that result in the release of chemicals into the environment are detrimental to male reproductive health. The reasons that these reference values will not fulfil these purposes are also explained. Although established reference values for semen characteristics are of limited value in analytical epidemiologic research, the WHO guidelines are of utmost importance for supporting the development of appropriate research protocols. Moreover, in spite of its limitations, semen analysis is still a useful research tool in epidemiological research, and no superior alternatives are on the horizon.

Practical semen analysis： from A to Z

Accurate semen analysis is critical for decisions about patient care, as well as for studies addressing overall changes in semen quality, contraceptive efficacy and effects of toxicant exposure. The standardization of semen analysis is very difficult for many reasons, including the use of subjective techniques with no standards for comparison, poor technician training, problems with proficiency testing and a reluctance to change techniques. The World Health Organization （WHO） Semen handbook （2010） offers a vastly improved set of standardized procedures, all at a level of detail that will preclude most misinterpretations. However, there is a limit to what can be learned from words and pictures alone. A WHO- produced DVD that offers complete demonstrations of each technique along with quality assurance standards for motility, morphology and concentration assessments would enhance the effectiveness of the manual. However, neither the manual nor a DVD will help unless there is general acknowledgement of the critical need to standardize techniques and rigorously pursue quality control to ensure that laboratories actually perform techniques ＇according to WHO＇ instead of merely reporting that they have done so. Unless improvements are made, patient results will continue to be compromised and comparison between studies and laboratories will have limited merit.

Quality analysis of commercial samples of Ziziphi spinosae semen(suanzaoren) by means of chromatographic fingerprinting assisted by principal component analysis

Due to the scarcity of resources of Ziziphi spinosae semen （ZSS）, many inferior goods and even adulterants are generally found in medicine markets. To strengthen the quality control, HPLC fingerprint common pattern established in this paper showed three main bioactive compounds in one chromatogram simultaneously. Principal component analysis based on DAD signals could discriminate adulterants and inferiorities. Principal component analysis indicated that all samples could be mainly regrouped into two main clusters according to the first principal component （PC1, redefined as Vicenin II） and the second principal component （PC2, redefined as zizyphusine）. PC1 and PC2 could explain 91.42%of the variance. Content of zizyphusine fluctuated more greatly than that of spinosin, and this result was also confirmed by the HPTLC result. Samples with low content of jujubosides and two common adulterants could not be used equivalently with authenticated ones in clinic, while one reference standard extract could substitute the crude drug in pharmaceutical production. Giving special consideration to the well-known bioactive saponins but with low response by end absorption, a fast and cheap HPTLC method for quality control of ZSS was developed and the result obtained was commensurate well with that of HPLC analysis. Samples having similar fingerprints to HPTLC common pattern targeting at saponins could be regarded as authenticated ones. This work provided a faster and cheaper way for quality control of ZSS and laid foundation for establishing a more effective quality control method for ZSS.

Semen analysis and sperm function testing

Despite controversy regarding the clinical value of semen analysis, male fertility investigation still relies on a standardized analysis of the semen parameters. This is especially true for infertility clinics in both developing and developed countries. Other optional tests or sophisticated technologies have not been widely applied. The current review addresses important changes in the analysis of semen as described in the new World Health Organization （WHO） manual for semen analysis. The most important change in the manual is the use of evidence-based publications as references to determine cutoff values for normality. Apart from the above mentioned changes, the initial evaluation and handling methods remain, in most instances, the same as in previous editions. Furthermore, the review evaluates the importance of quality control in andrology with emphasis on the evaluation of sperm morphology. WHO sperm morphology training programmes for Sub-Saharan countries were initiated at Tygerberg Hospital in 1995. The external qualitY control programme has ensured that the majority of participants have maintained their morphological reading skills acquired during initial training. This review reports on current sperm functional tests, such as the induced acrosome reaction, and sperm-zona pellucida binding assays, as well as the impact of sperm quality in terms of DNA integrity, and the relationship of sperm function tests to sperm morphology.

Semen analysis standardization： is there any problem in Polish laboratories？

The aim of the study was to determine the degree of compliance of Polish laboratories with World Health Organization （WHO） recommendations, with regard to semen analysis methodology. A survey requesting information about methods of semen analysis was distributed to employees of 55 laboratories. Respondents who had participated in external seminological workshops （31%） were termed certified respondents （CR）, the remaining （69%）--non-certified respondents （NCR）. Only one laboratory （6%） in the CR group and none in the NCR were compliant with WHO guidelines for methods and equipment used to evaluate seminal volume, sperm motility, concentration, vitality and morphology. Most problems were of volume measurement （weighing method was reported by 17% of CR and 10% of NCR） and staining method for sperm morphology （Papanicolau or Diff-Quik were found in 33% of CR and 23% of NCR）. A three- or four-point grading of sperm motility was used by the majority of respondents; however, 17% of CR and 37% of NCR did not use a laboratory counter to tally spermatozoa. Although a haemocytometer method was used by 80% of laboratories in each group, the improved Neubauer chamber was used only by 42% of CR and 19% of NCR. In each group, 24% of laboratories did not perform a vitality test. Procedural errors and the interchangeable utilization of two or even three methods to analyse a given parameter was observed in both groups. The results indicate a need for standardisation of the methods and continuous, unified training in semen analysis in Polish laboratories.

A comparison of conventional and computer-assisted semen analysis.（CRISMAS software） using samples from 166 young Danish men

The aim of the present study was to compare assessments of sperm concentration and sperm motility analysed by conventional semen analysis with those obtained by computer-assisted semen analysis （CASA） （Copenhagen Rigshospitalet Image House Sperm Motility Analysis System （CRISMAS） 4.6 software） using semen samples from 166 young Danish men. The CRISMAS software identifies sperm concentration and classifies spermatozoa into three motility categories. To enable comparison of the two methods, the four motility stages obtained by conventional semen analysis were, based on their velocity classifications, divided into three stages, comparable to the three CRISMAS motility categories： rapidly progressive （A）, slowly progressive （B） and non-progressive （C＋ D）. Differences between the two methods were large for all investigated parameters （P〈0.001）. CRISMAS overestimated sperm concentration and the proportion of rapidly progressive spermatozoa and, consequently, underestimated the percentages of slowly progressive and non-progressive spermatozoa, compared to the conventional method. To investigate whether results drifted according to time of semen analysis, results were pooled into quarters according to date of semen analysis. CRISMAS motility results appeared more stable over time compared to the conventional analysis; however, neither method showed any trends. Apparently, CRISMAS CASA results and results from the conventional method were not comparable with respect to sperm concentration and motility analysis. This needs to be accounted for in clinics using this software and in studies of determinants of these semen characteristics.

Semen analysis with regard to and functional aspects sperm number, sperm morphology

The new World Health Organization （WHO） Manual for Semen Analysis contains several improvements. One is that the 20 million spermatozoa per mL paradigm has been ousted in favour of proper calculations of lower reference limits for semen from men, whose partners had a time-to-pregnancy of 12 months or less. The recommendation to grade the progressive motility as described in the third and fourth editions of the WHO manual was not evidence-based, and WHO was therefore motivated to abandon it. However, the new recommendation is not evidence-based either, and it is difficult to understand the rational for the new assessment. It may have been a compromise to avoid returning to the rather robust system recommended in the first edition （1980）. The unconditional recommendation of the ＇Tygerberg strict criteria＇ is not evidence-based, and seems to be the result of an unfortunate bias in the composition of the Committee in favour of individuals known to support the ＇strict criteria＇ method. This recommendation will have negative effects on the develop- ment ofandrology as a scientific field. Given the importance of the WHO manual, it is unfortunate that the recommenda- tions for such important variables, as motility and morphology, lack evidence-based support.

Semen analysis in chronic bacterial prostatitis： diagnostic and therapeutic implications

The significance and diagnostic value of semen analysis in chronic bacterial prostatitis has been extensively debated and remains controversial. To investigate the diagnostic relevance of semen culture in the bacteriological workup of prostatitis patients, we retrospectively analyzed a clinical database of 696 symptomatic patients. All patients were routinely subjected to a four-glass test, followed by semen culture and analysis. This allowed to dissect from the database three different diagnostic scenarios, and to compare the ＇two-glass＇ pre-/post- massage test and the standard ＇four-glass＇ test with a ＇five-glass＇ test （four-glass plus post-VB3 semen culture）. The ＇five-glass＇ test showed 3.6- or 6.5-fold increases in relative sensitivity and lesser reductions （-13.2% or -14.7%） in relative specificity for traditional uropathogens （TUs） compared with the four-glass or two-glass test, respectively. The area under the ROC curve and Jouden＇s index were increased, whereas positive and negative likelihood ratios were lower than comparators, indicating that the ＇five-glass＇ assay may be superior in confirming the negative outcome of both standard tests. The five-, four-, and two-glass tests detected TUs （Enterobacteriaceae, Enterococci, etc.） in 120, 33, and 20 patients and unusual pathogens （Streptococci, other Gram-positive species, Mycoplasmata, and others） in 130, 56, and 45 patients, respectively. When patients were subjected to pharmacological treatment, including a combination of a fluoroquinolone and a macrolide, no differences in eradication rates were observed between groups diagnosed with different tests, irrespective of pathogen category. Eradication was associated with long-term sign/symptom remission; no significant intergroup differences in sign/symptom scores were observed throughout a 24-month off-therapy follow-up period. In conclusion, our data support the usefulness of semen analysis in the diagnostic workup ofprostatitis patients when this test is used to complement the four-glass Meares and Stamey test. Improvement of microbiological assays conveys important diagnostic and therapeutic implications.

Semen Analysis and Fecundity Association Among Women with Polycystic Ovary Syndrome Experiencing Ovulatory Dysfunction Treated by Ovulation Induction

In this study,normal values of semen analysis were set for a general infertile population of couples among which most women had normal ovulation.The predictive capacity values of sperm quality,including concentration,motile count,progressive motile count,and morphology,are unclear for women with polycystic ovary syndrome(PCOS).A secondary analysis was conducted based on a randomized controlled trial investigating infertility among women with PCOS experiencing ovulatory disorder between 2011 and 2016 in China.A total of 1000 women received ovulation induction(acupuncture and clomiphene).We randomized the women with PCOS in 27 hospitals in China who received one of four interventions(acupuncture plus clomiphene,sham acupuncture plus clomiphene,acupuncture plus placebo,or sham acupuncture plus placebo).Semen analysis was performed for every male partner according to the World Health Organization(WHO)criteria.The outcomes included conception,clinical pregnancy,and live birth.Logistic regression was used to evaluate the predictive value of semen analysis among ovulatory women for conception,clinical pregnancy,and live birth.Among the 1000 couples,the number of couples who attained ovulation,conception,clinical pregnancy,and live birth were 780,320,235,and 205,respectively.Semen volume and motility were applied and used as prediction parameters for conception(area under the curve(AUC)of 0.62(95%confidence interval(CI),0.55–0.69)),clinical pregnancy(AUC of 0.67(95%CI:0.61–0.73)),and live birth(AUC of 0.57(95%CI:0.50–0.64)).No poor calibration was shown for these models in Hosmer–Lemeshow tests.The predictive capacity of semen analysis for treatment outcome in PCOS women with PCOS experiencing with ovulatory dysfunction is limited.

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 %) as compared to the combined categories of slow and rapid linear progressive. Conclusion: The present study has confirmed similar findings from other different countries that declining sperm counts in humans is not a global phenomenon.

Determination of the diagnostic value of the resazurin reduction assay for evaluating boar semen by receiver operating characteristic analysis

Aim： To assess that metabolic status of spermatozoa could provide a useful tool for evaluation of semen quality. Methods： The accuracy of the spectrophotometric application of the resazurin reduction assay was assessed using receiver operating characteristic （ROC） analysis. Results： Areas under ROC curves （AUC） for motile sperm concentration and sperm index （SI） （sperm concentration multiplied by the square root of percentage sperm motility multiplied by the percentage normal sperm morphology） were 0.922. The best discrimination between poor and good semen samples according to the SI was achieved at a cut-off point of A610 = 0.209, where high sensitivity （94.1%） and specificity （91.7%） were calculated. The assay was less accurate when motile sperm concentration was used as the criterion value, yielding sensitivity of 88.2% and specificity of 87.5%, respectively. Likelihood ratios （LR） indicate that absorbances lower than 0.209 were at least 11.3 times as likely to be found in good semen samples than those in poor according to the SI, whereas in the case of motile sperm concentration, the LR was calculated to be 7.06. Conclusion： These results show that the resazurin reduction assay combined with spectrophotometry is an accurate method of assessing the quality of boar semen.

Principal Component Analysis on Semen Quality among Chinese Young Men

Objective To understand the current semen quality status among Chinese young men and influential factors in China and to explore its evaluation index. Methods A total of 562 healthy male volunteers were recruited during their premarital examinations in seven provincials and municipal regions' MCH centers; descriptive and principal component analyses were used to analyze data.Results The findings show that semen volume (2. 61± 1. 10 mL), sperm density (64. 47× 34. 59× 106/mL), percentage of sperm forward progression (59. 89%± 17. 11%), percentage of sperm viability (77. 19% ± 11. 87%), and percentage of normal sperm morphology ( 78. 23% ± 9. 15% ). The first principal component function is Z1= -8.512 54 + 0. 001 35X1' + 0. 031 92X2'+0. 043 52X3'+ 0. 039 84X4', which is closely related to percentage of sperm viability (X3), percentage of sperm forward progression (X2), and percentage of normal sperm morphology (X4)The second principal component function is: Z2= 0. 491 92+ 0. 080 80X1- 0. 000 58X2-0. 005 10X3- 0. 018 07X4, which depends on the total sperm count (X1). Conclusion Only 42. 3% subjects meet all the common WHO standard of semen quality. The multiple analysis of Z1 showed that the highest Z1 are among subjects from Guizhou,workers, or town residents. Multiple analysis of Z2 showed that the older age when the subjects had the first sexual impulse, the longer period of sexual abstinence and more quantity of sperm they had; the more sexual activity subjects had, the less amount of sperm they had.

Determination of the diagnostic value of the resazurin reduction assay for evaluating boar semen by receiver operating cha-racteristic analysis

Aim:To assess that metabolic status of spermatozoa could provide a useful tool for evaluation of semen quality. Methods:The accuracy of the spectrophotometric application of the resazurm reduction assay was assessed using receiver operating characteristic(ROC)analysis.Results:Areas under ROC curves(AUC)for motile sperm concen- tration and sperm index(SI)(sperm concentration multiplied by the square root of percentage sperm motility multi- plied by the percentage normal sperm morphology)were 0.922.The best discrimination between poor and good semen samples according to the SI was achieved at a cut-off point of A_(610)=0.209,where high sensitivity(94.1%)and specificity(91.7%)were calculated.The assay was less accurate when motile sperm concentration was used as the criterion value,vielding sensitivity of 88.2% and specificity of 87.5%,respectively,Likelihood ratios(LR)indicate that absorbances lower than 0.209 were at least 11.3 times as likely to be found in good semen samples than those in poor according to the SI,whereas in the case of motile sperm concentration,the LR was calculated to be 7.06.Conclusion: These results show that the resazurin reduction assay combined with spectrophotometry is an accurate method of assessing the quality of boar semen.

Impact of chronological ageing on semen parameters in southern Indian men visiting infertility centre:A retrospective study

Objective:To investigate the association between age and semen parameters among male partners of subfertile couples.Methods:This retrospective study analyzed the semen of 1523 infertile men aged 26 to 50 years.Data were extracted from GarbhaGudi IVF Centre database from January 2019 to September 2020.The basic semen parameters were interpreted according to the WHO manual 2021,6th edition.Semen parameters in different age groups were compared.Results:Total and progressive motile sperms were significantly higher in the age group of 26-30 years compared to other age groups(P<0.05).Normal sperm count was significantly higher in the age group of 26-30 years compared to the age groups of 41-45 years and>46 years(P=0.001).However,sperm head defects,neck and midpiece defects,tail defects,and cytoplasmic droplets showed statistically insignificant difference in all the age groups(P>0.05).Semen viscosity showed no statistical difference in all the age groups compared to the reference age group of 26 to 30 years.Conclusions:Higher age can lead to a significant decrease in normal sperms and motility in subfertile men.Hence,male partner age should be considered as one of the major determining factors for reproductive outcomes.

Male Infertility as a Circumstance of Discovery of Adult Cryptorchidism: A Retrospective Analysis from the Region of Thies

Introduction: The aim of this work is to study the clinical aspects and the impact of cryptorchidism on male infertility in the region of Thies. Patients and methods: This is a retrospective study involving 28 patients followed up at the Saint Jean de Dieu, Barthimée and Tivaouane hospitals of the region of Thies between January 2007 and December 2016. Results: Among the 223 patients followed up for the undescended testicle, 13% (n = 28) were adults or at least 15 years old. 71% (n = 20) were married. Fourteen patients were followed up for primary infertility lasting for a period of time averaging 7 years. The mean age of the patients was 28 years (range: 17 years and 45 years). Cryptorchidism was unilateral in 82% (n = 23) and bilateral in 18% (n = 5). The mean delay of consultation varied from 1 to 13 years. The main reason for consultation was couple infertility (50%). The semen analysis was requested in all our patients. The levels of FSH, LH and testosterone were measured by immunoassay for the patients with azoospermia (n = 9). We noticed 2 cases of hypotestosteronemia and 3 cases of increased FSH serum level. Computed Tomography scans (CT) were performed in 28.5% of patients (n = 8) and ectopic testicles were found very high in the inguinal area in 5 cases. The open inguinal approach was used. In perioperative period, the testicle was found, high in the inguinal region in 5 patients (17.8% of the cases) and low in inguinal region in 19 patients (67.8%). In 4 cases the testicle was in abdominal position. The testicle was atrophied in 53.5% of the patients (n = 15) and 8 patients presented testicular hypotrophy. An orchidectomy was performed in 1 patient in whom it was impossible to lower the atrophied testicle. The postoperative period was uneventful and simple. The mean follow-up duration after the operation was 36 months (12 - 60 months).

国家药品计划抽检品种麸炒薏苡仁质量问题分析及评价

目的:通过国家药品计划抽检工作,对药食两用中药材麸炒薏苡仁的质量进行考察,了解麸炒薏苡仁饮片质量现状及存在的问题,完善麸炒薏苡仁检验标准,为药品监管部门提供技术支持。方法:从全国24个省(市、自治区)的生产和使用环节抽取麸炒薏苡仁样品,按照《中华人民共和国药典》2020年版一部进行性状、水分、灰分、浸出物及含量测定项检验,分析检验数据以评价麸炒薏苡仁饮片的质量状况,并针对检验中发现的问题,开展探索性研究。结果与结论:本次抽验的麸炒薏苡仁样品共计98批次,按现行质量标准检验合格率均为100%。经探索性研究,发现麸炒薏苡仁存在炮制不到位的问题,33批次样品中4批次玉米赤霉烯酮超标;此外,甘油三油酸酯含量测定项限度偏低。通过探索性研究发现麸炒薏苡仁饮片和标准中的问题,结合标准提高工作完善麸炒薏苡仁检验标准。建议增加安全性控制指标及炮制控制指标,并提高含量测定限度。