World Health Organization Laboratory Manual Research Articles

Sixth edition of the World Health Organization laboratory manual of semen analysis: Updates and essential take away for busy clinicians

ABSTRACT Background Semen analysis is the cornerstone for male fertility evaluation. In 2021, the World Health Organization (WHO) released its 6th edition of semen analysis manual. Methods We highlight the main changes in the latest 6th edition of the WHO manual of semen analysis and their possible interpretations. Results The manual is highly comprehensive, offering detailed information, and is widely regarded as an excellent technical reference for laboratory staff. Nevertheless, several aspects of the manual require further elucidation for infertility practitioners. Conclusion The recently published 6th edition of the WHO manual provides a strong framework for the assessment and processing of human semen. Grasping the modifications introduced in this updated edition and their clinical significance can enhance the quality of patient care.

O-285 The 6th edition of the WHO manual for the examination and processing of human semen: How should it affect our practice?

Abstract The WHO laboratory manual (World Health Organization 2021) has widened the scope of semen examination from primarily a prognostic tool for Medically Assisted Reproduction (MAR) by indicating the importance of evaluating men with possible disorders in male reproductive functions. Practical instructions for basic examination have been clarified and focused to measures of sperm production and motility as judged from sperm number, motility (vitality if poor motility) and sperm morphology. In addition to the recommendations by the WHO, an international standard based on the same laboratory science as the WHO manual has been published (International Organization for Standardization 2021) to facilitate for laboratories to provide reliable results of semen examination. Each ejaculate must be handled with great care before analysis begins. Time and temperature between sample collection and start of assessments is crucial. To reduce assessment variability, it is essential to ascertain that the examined aliquots are representative for the entire ejaculate and that the number of observations is sufficient to reduce the risk for significant influence of random factors. The former is handled both by aliquot volume (at least 50 µL for sperm concentration) and replicate assessment and comparison that the two assessments don’t differ too much. Regarding number of observations, 400 sperm counted in sperm concentration and in sperm motility assessments is required to reduce this source of errors to ± 10%. For assessment of sperm morphology and vitality a minimum number of observations is 200 spermatozoa. To achieve acceptable performance, thorough in-house training is necessary. There are protocols how to set up and run such schemes (Mortimer 1994, 1994). Basic medical laboratory standards also require that the laboratory regularly performs internal quality control to monitor that inter- and intra-personal variability is under control. Each laboratory should also participate in an external scheme for quality assessment (EQA) (International Organization for Standardization 2012). For the proper interpretation of semen examination results reference limits are important. However, the reference limits suggested by the WHO (World Health Organization 2021) come from a very mixed group of men and should therefore not be mistaken for true limits between fertility and infertility. The new WHO manual therefore argues for development of decision limits (when is it reasonable to act?) that are much more important than limits from a mixed population. It has been argued that semen examination must be better standardized (Björndahl et al. 2016), but the compliance has been almost non-existent (Vasconcelos et al. 2022) leading to even stronger appeals for stronger improvement of basic laboratory investigations of male reproductive functions (Björndahl et al. 2022) References Björndahl L, Barratt CL, Mortimer D, and Jouannet P. ‘How to count sperm properly': checklist for acceptability of studies based on human semen analysis. Hum Reprod 2016: 31; 227-232. Björndahl L, Barratt CLR, Mortimer D, Agarwal A, Aitken RJ, Alvarez JG, Aneck-Hahn N, Arver S, Baldi E, Bassas L, et al. Standards in semen examination: publishing reproducible and reliable data based on high-quality methodology. Hum Reprod 2022. International Organization for Standardization. ISO 15189:2012 Medical Laboratories – Requirements for Quality and Competence 2012. Geneva. International Organization for Standardization. ISO 23162:2021 Basic semen examination — Specification and test methods 2021. ISO, Geneva. Mortimer D. Laboratory standards in routine clinical andrology. Reproductive Medicine Review 1994: 3; 97-111. Mortimer D. Practical Laboratory Andrology 1994. Oxford University Press, Oxford. Vasconcelos AL, Campbell MJ, Barratt CLR, and Gellatly SA. Do studies published in two leading reproduction journals between 2011 and 2020 demonstrate that they followed WHO5 recommendations for basic semen analysis? Hum Reprod 2022. World Health Organization. WHO laboratory manual for the examination and processing of human semen. 6th edn, 2021. World Health Organization, Geneva.

Association between central obesity and semen quality: A cross-sectional study in 4513 Chinese sperm donation volunteers.

General obesity classified by body mass index has been linked to a reduction in semen quality; however, evidence on the adverse effect of central obesity on semen quality remains limited. To investigate the association between central obesity and semen quality. We conducted a cross-sectional study of 4513 sperm donation volunteers in Guangdong Provincial Human Sperm Bank during 2018-2021. Three central obesity indicators, including waist circumference, waist-to-hip ratio, and waist-to-height ratio, were measured using a multi-frequency bioelectrical impedance analysis for each subject. Semen analysis was conducted according to the World Health Organization laboratory manual for the examination and processing of human semen 5th edition. Linear regression models and unconditional logistic regression models were used to quantify the association between central obesity and semen parameters. With adjustment for age, race, education level, marital status, fertility status, occupation, year of semen collection, abstinence period, ambient temperature, and relative humidity, central obesity defined as waist circumference ≥90cm, waist-to-hip ratio ≥0.9, or waist-to-height ratio ≥0.5 was significantly associated with a 0.27 (95% confidence interval: 0.15, 0.38) mL, 14.47 (3.60, 25.34) × 106 , 7.06 (0.46, 13.76)× 106 , and 6.80 (0.42, 13.18) × 106 reduction in semen volume, total sperm number, total motile sperm number, and total progressive motile sperm number, respectively, and a 53% (10%, 112%) increase in odds of below the World Health Organization 2010 reference value for semen volume. These associations did not significantly vary across age. Similar results were observed for central obesity defined using each of the three indicators, except that subjects with a waist circumference ≥90cm had a slightly higher total motility (estimated change: 1.30%; 95% confidence interval: 0.27%, 2.34%) and progressive motility (estimated change: 1.27%; 95% confidence interval: 0.23%, 2.31%). We found that central obesity was significantly associated with a reduction in semen volume, total sperm number, total motile sperm number, and total progressive motile sperm number. Future studies are warranted to confirm our results in other regions and populations.

Dietary total antioxidant capacity and the risk of developing asthenozoospermia: a hospital-based case-control study in China.

Is dietary total antioxidant capacity (DTAC) associated with the odds of developing asthenozoospermia in Chinese men? There is no statistically significant association between DTAC indices and the odds of developing asthenozoospermia. Both diet and oxidative stress may be related to sperm quality; however, few studies have investigated the association between DTAC and sperm quality. This case-control study was conducted from June 2020 to December 2020. Those diagnosed with asthenozoospermia were assigned to the case group, whereas those with normal sperm parameters were assigned to the control group. Data from a total of 553 cases and 586 controls were included in the final analysis. Men who had been referred to the infertility clinic of Shengjing Hospital of China Medical University were enrolled. Dietary intake was assessed using a validated food frequency questionnaire. DTAC was based on ferric-reducing ability of plasma (FRAP), total oxygen radical absorbance capacity (T-ORAC), hydrophilic oxygen radical absorbance capacity (H-ORAC), lipophilic oxygen radical absorbance capacity (L-ORAC), total phenolics (TP), total radical-trapping antioxidant parameter (TRAP), and Trolox equivalent antioxidant capacity (TEAC). Asthenozoospermia was defined according to the criteria published in the fifth edition of the World Health Organization laboratory manual for the examination and processing of human semen. No significant association was observed between the DTAC indices and the odds of asthenozoospermia after multivariable adjustment (T3 vs T1, odds ratio (OR) = 0.99, 95% CI: 0.73-1.33 for FRAP; OR = 1.05, 95% CI: 0.77-1.42 for T-ORAC; OR = 0.88, 95% CI: 0.65-1.18 for H-ORAC; OR = 0.98, 95% CI: 0.71-1.34 for L-ORAC; OR = 1.03, 95% CI: 0.76-1.39 for TP; OR = 1.18, 95% CI: 0.87-1.59 for TRAP; and OR = 1.15, 95% CI: 0.85-1.55 for TEAC). Both additive and multiplicative interaction analyses suggested that smoking might modify the association of T-ORAC with the odds of developing asthenozoospermia (relative excess risk due to interaction = 0.45, 95% CI: 0.07-0.83, attributable proportion due to interaction = 0.46, 95% CI: 0.07-0.84 for additive interaction; P = 0.033 for multiplicative interaction). Recall bias and protopathic bias were inevitable in this retrospective case-control study. The estimation accuracy of the DTAC indices may have also affected the findings. To the best of our knowledge, this is the first study to specifically investigate whether an association exists between DTAC and the odds of developing asthenozoospermia. Although no significant association was found, this study provides novel information pertaining to the fields of nutrition and human reproduction. This work was supported by the JieBangGuaShuai Project of Liaoning Province (2021JH1/10400050), the Shengjing Hospital Clinical Research Project (M0071), and the Outstanding Scientific Fund of Shengjing Hospital (M1150). All authors have no competing interests to disclose. N/A.

Controversy and Consensus on Indications for Sperm DNA Fragmentation Testing in Male Infertility: A Global Survey, Current Guidelines, and Expert Recommendations.

Sperm DNA fragmentation (SDF) testing was recently added to the sixth edition of the World Health Organization laboratory manual for the examination and processing of human semen. Many conditions and risk factors have been associated with elevated SDF; therefore, it is important to identify the population of infertile men who might benefit from this test. The purpose of this study was to investigate global practices related to indications for SDF testing, compare the relevant professional society guideline recommendations, and provide expert recommendations. Clinicians managing male infertility were invited to take part in a global online survey on SDF clinical practices. This was conducted following the CHERRIES checklist criteria. The responses were compared to professional society guideline recommendations related to SDF and the appropriate available evidence. Expert recommendations on indications for SDF testing were then formulated, and the Delphi method was used to reach consensus. The survey was completed by 436 experts from 55 countries. Almost 75% of respondents test for SDF in all or some men with unexplained or idiopathic infertility, 39% order it routinely in the work-up of recurrent pregnancy loss (RPL), and 62.2% investigate SDF in smokers. While 47% of reproductive urologists test SDF to support the decision for varicocele repair surgery when conventional semen parameters are normal, significantly fewer general urologists (23%; p=0.008) do the same. Nearly 70% would assess SDF before assisted reproductive technologies (ART), either always or for certain conditions. Recurrent ART failure is a common indication for SDF testing. Very few society recommendations were found regarding SDF testing. This article presents the largest global survey on the indications for SDF testing in infertile men, and demonstrates diverse practices. Furthermore, it highlights the paucity of professional society guideline recommendations. Expert recommendations are proposed to help guide clinicians.

The association between healthy eating index score with semen parameters in infertile men: A cross-sectional study.

Infertility has been a major problem for young couples in recent years. One way to assay the diet quality is the healthy eating index (HEI), related to infertility. This study aims to assess the association between the HEI score with semen parameters in Iranian infertile men. Two hundred and sixty eligible men (18-55 yr), were referred to the major infertility clinic in the summer of 2018 and entered this cross-sectional study. Based on the 5 edition of the world health organization laboratory manual, semen parameters including sperm concentration, volume, motility, and morphology were analyzed, and to specify the dietary intake of individuals a 168-item questionnaire was used. Also, to calculate the total HEI score, all 13 components based on HEI-2015 components and scoring standards were summed up. Participants in the highest tertile, had no difference in mean sperm parameters with those in the lowest tertile in the crude model. No significant association was found between sperm parameters and HEI score tertiles in the crude model, even after adjustment for potential confounders, except for concentration (OR: 0.39 and CI: 0.15, 0.99, p = 0.04). Participants in the highest tertile had a lower risk of abnormal concentration and motility in the crude model. The risk of abnormal concentration decreased, and motility increased in the adjusted model. In this cross-sectional study, there was no significant relationship between HEI and sperm indexes, except for sperm concentration. Therefore, more studies need to be done in the future.

Do studies published in two leading reproduction journals between 2011 and 2020 demonstrate that they followed WHO5 recommendations for basic semen analysis?

STUDY QUESTIONDo publications that involve the interpretation of the results of a basic semen analysis, published in Human Reproduction and Fertility & Sterility between 2011 and 2020, give sufficient evidence in their methodology to demonstrate that they followed the technical methods recommended in the fifth edition of the World Health Organization (WHO) laboratory manual, entitled WHO Laboratory Manual for the Examination and Processing of Human Semen (WHO5)?SUMMARY ANSWEREvidence of methodological agreement of studies with the WHO5 recommendations was low, despite 70% of papers stating that they followed WHO5 recommendations.WHAT IS KNOWN ALREADYA basic semen analysis is currently an integral part of infertility investigations of the male, but method standardization in laboratories remains an issue. The different editions of the WHO manual for the basic semen analysis (WHO1–6) have attempted to address this by providing increasingly rigorous methodological protocols to reduce experimental error. However, to what extent these methods are followed by studies that involve the interpretation of the results of basic semen analysis remains unknown.STUDY DESIGN, SIZE, DURATIONA survey of the technical methods used to perform a basic semen analysis was conducted on studies published in two leading reproduction journals (Human Reproduction and Fertility & Sterility) between 2011 and 2020.PARTICIPANTS/MATERIALS, SETTING, METHODSThe literature search was performed on the electronic databases PUBMED and MEDLINE Ovid between January 2021 and March 2021. The MeSH terms included in the search were ‘sperm concentration’ OR ‘sperm motility’ OR ‘sperm morphology’ OR ‘sperm vitality’ OR ‘male fertility’ AND ‘human spermatozoa’ NOT ‘animals’. A total of 122 studies were available for analysis.MAIN RESULTS AND THE ROLE OF CHANCEIn total, 70% of the studies cited WHO5 in their methods section. Of the remaining studies, 10% cited the fourth edition of the WHO laboratory manual (WHO4), 7% cited both WHO4 and WHO5, 1% cited the third edition of the WHO laboratory manual (WHO3), and 12% did not cite the WHO at all. Overall methodological agreement with WHO5 recommendations was poor, with the main reason for this lack of agreement being that the research studies did not disclose specific details of the technical methods and equipment used.LIMITATIONS, REASONS FOR CAUTIONIn the case of studies that did not disclose any specific technical methods that they used, we did not attempt to contact these authors and so were unable to confirm the agreement between their technical methods and WHO5 recommendations.WIDER IMPLICATIONS OF THE FINDINGSOur findings suggest there is an urgent need to develop strategies to address standardization in reporting the results of a semen analysis for publication. This is particularly timely given the recent publication of WHO6 and ISO standard 23162 for the basic examination of human semen.STUDY FUNDING/COMPETING INTEREST(S)There was no funding for this project. C.L.R.B., as an employee of the University of Dundee, serves on the Scientific Advisory board of ExSeed Health (from October 2021, financial compensation to the University of Dundee) and is a scientific consultant for Exscientia (from September 2021, financial compensation to the University of Dundee). C.L.R.B. has previously received a fee from Cooper Surgical for lectures on scientific research methods outside the submitted work (2020) and Ferring for a lecture on male reproductive health (2021). C.L.R.B. is Editor for RBMO.TRIAL REGISTRATION NUMBERN/A.

The past, present, and future of the semen analysis

This month's Views and Reviews provides an added perspective to the World Health Organization laboratory manual for the examination and processing of human semen, which was recently published in the 6th edition. The first artice provides a historical context of the prior editions of the World Health Organization manuals and modifications adopted over the years. The next piece then provides additional perspectives on the methodologies used for the performance of semen analysis. The third article then examines some of the new semen analytic technologies and enhancements that have become more common over the years. Finally, the last article proposed where male reproductive testing will head in the coming years with emerging research and technologies.

Pattern of semen analysis in male partners of infertile couples in Western Ethiopia: Retrospective cross-sectional study.

Objectives:This study assesses the pattern of semen analysis results in male partners of infertile couples at Gimbie Adventist Hospital, Western Ethiopia, 2021.Methods:A retrospective cross-sectional study on 131 semen samples of male partners of infertile couples was conducted at Gimbie Adventist Hospital from 5 September 2021 to 5 October 2021. All semen samples were processed and analyzed according to methods and standards outlined by the World Health Organization laboratory manual for the examination and processing of human semen 2010. The data were coded and entered into EpiData version 3.1, and then cleaned and exported to Statistical Package for Social Sciences (SPSS for Windows version 25) for analysis. The results were presented in tables, figures, and charts.Results:The age of study participants ranges from 20 to 65 years with a mean age of 30.2 ± 8.1 years. Sperm cell count, morphology, total motility, and vitality below the World Health Organization reference level were found in 48.9%, 27.5%, 43.5%, and 67.2% of the analyzed samples, respectively. Low power of hydrogen and high viscosity were observed in 31.3% and 16.8% of the semen samples, respectively. The majority, 84%, had one or more abnormal semen analysis parameters. Asthenozoospermia (43.5%), necrozoospermia (25.2%), oligozoospermia (24%), azoospermia (24%), and oligoasthenoteratozoospermia (25.2%) were the severe forms of abnormal semen analysis findings detected in this study. The decline in sperm cell morphology and motility were noticed after the age of 31–34 years.Conclusion:In this study, both sperm quantity and quality were more affected when compared to similar studies. Only 16% of analyzed samples had normal semen parameters. Given this finding, identifying risk factors and introducing advanced diagnostic modalities for the workup of male infertility in the study area are highly recommended.

RELATIONSHIP OF SEMINAL FRUCTOSE AND SERUM PROLACTIN LEVELS IN INFERTILE MEN

Objective: Male infertility refers to a male’s inability to cause pregnancy in a fertile female. Infertility is a widespread problem for about one in five infertile couples. Male infertility is usually caused by a problem that affects either sperm production or sperm transport. The aim of the study was to evaluate prolactin levels in the serum and comparing the levels of fructose in the semen of infertile males and normal healthy controls of reproductive age of 20–45 years respectively. Methods: Hemoglobin (Hb) level in the blood of both groups was calculated by Sahli’s method using Sahli’s Hemoglobinometer. The Erythrocyte Sedimentation Rate (ESR) was determined using the modified Westergren’s method and total leukocyte count (TLC) in the blood was determined by microdilution method. Semen analysis was performed by the method specified in the World Health Organization Laboratory Manual. Morphology and motility of sperm and their counts were determined by microscopy. Fructose drop test in semen was estimated colorimetrically by the Semen Fructose Estimation Kit and determination of serum prolactin involved the use of Electrochemiluminescence immunoassay using Cobas e 411 analyzer. Results: Hb in infertile males was less than healthy controls. However, ESR in infertile men was considerably high and TLC, serum prolactin levels did not show much variation in infertile men. Seminal analysis of infertile men revealed high fructose levels and low sperm count with decreased motility. Conclusion: The concentration of seminal fructose in control group is lower than infertile males. High fructose levels in infertile men and less sperm count indicate oligozoospermia.

Associations between depression, oxidative stress, and semen quality among 1,000 healthy men screened as potential sperm donors

To explore the association between depression and semen quality and the mediating role of oxidative stress. Cross-sectional study with repeated measures of semen quality. Human Sperm Bank of Hubei Province, People's Republic of China. From April 2017 to July 2018, we recruited 1,000 potential sperm donors who completed the Beck Depression Inventory questionnaire and had measures of oxidative stress biomarkers. None. Severity of depression was evaluated by the Beck Depression Inventory scores (0-4, no depression; 5-13, mild depression; 14-20, moderate depression; and 21 or greater, severe depression). The urinary concentrations of 8-hydroxy-2-deoxyguanosine, 4-hydroxy-2-nonenal-mercapturic acid, and 8-iso-prostaglandin F2α (8-isoPGF2α) were measured to reflect oxidative stress status. Repeated semen quality parameters (n = 5,880) were examined by trained professional technicians according to the World Health Organization laboratory manual. Associations between depression, oxidative stress, and repeated measures of semen quality parameters were evaluated using linear or mixed-effects models with adjustment for potential confounders. Mediation analysis was performed to test the potential mediating role of oxidative stress. A total of 391 (39.1%) men were classified as mild depression, 67 (6.7%) as moderate depression, and 19 (1.9%) as severe depression. Inverse dose-response relationships between severity of depression and semen quality parameters were found. Compared with men without depression (n = 523), those with severe depression had a 25.26% (95% confidence interval, -38.65%, -8.93%) lower semen volume, 37.04% (-55.37%, -11.20%) lower total sperm count, 13.57% (-23.17%, -2.78%) lower total motility, and 15.08% (-25.09%, -3.72%) lower progressive motility; men with moderate depression also had a 12.28% (-21.16%, -2.40%) lower semen volume and 23.56% (-36.50%, -7.97%) lower total sperm count. We found a positive dose-response relationship between severity of depression and urinary 8-isoPGF2α concentrations. However, we found no evidence that the associations between depression status and semen quality were mediated by oxidative stress markers. In the study of Chinese male sperm donors, men with depression had worse semen quality parameters, including semen volume, sperm concentration, total sperm count, total motility, and progressive motility. Although depression was positively associated with urinary 8-isoPGF2α concentrations, depression-semen quality associations were not mediated by oxidative stress.

The Effect of Ejaculatory Abstinence Interval on Sperm Parameters and Clinical Outcome of ART. A Systematic Review of the Literature.

Since the publication of the first edition of the WHO (World Health Organization) Laboratory Manual for the examination of Human Semen in 1980, the reference values of sperm parameters have been updated on four occasions. Currently and globally, most of the laboratories analyzing semen samples use the latest, 5th edition of the manual that recommends ejaculatory abstinence from two to seven days before producing the sample for examination. While this standardized interval of time facilitates the interpretation of the results and research, no solid evidence exists to support the WHO-recommended abstinence time for a semen analysis in order to optimize clinical outcomes after assisted reproduction. Most of the studies refer to different clinical outcomes, different groups of patients and different editions of the WHO Laboratory Manual, including heterogeneous intervals of abstinence or sperm parameters. The aim of the current systematic review was to evaluate available evidence correlating ejaculatory abstinence time with clinical outcomes and sperm parameters analyzed according to the last edition of the World Health Organization Laboratory Manual reference values in different male populations. The results from the included studies indicate that WHO abstinence recommendations may need revision, given that a shorter ejaculatory abstinence interval appears to be associated with improved sperm parameters, such as sperm DNA fragmentation, progressive motility or morphology, while evidence suggests a potential increase in embryo euploidy rates and pregnancy outcomes.

Study of Semen Analysis Patterns in Male Partner of Infertile Couple Attending Tertiary Level Hospital of Nepal

Aims: The aim of this study is to analyze the pattern of semen abnormality in male partner of infertile couple in Nepal. Methods: A retrospective study of semen sample of male partner of infertile couple analyzed in Department of pathology, Dhulikhel Hospital from January 2014 to December 2018. All semen samples were processed and analyzed according to methods and standards outlined by World Health Organization laboratory manual for the examination and processing of human semen 2010. Results: A total of 520 semen samples were analyzed. Our study shows 221 (44%) abnormal for different semen parameters and asthenozoospermia (39.3%) is the most common abnormality followed by azoospermia (28.8 %), Oligoasthenozoospermia (17.9 %), Oligozoospermia (8.7 %), Oligoasthenoteratozoospermia (3.5 %) and teratozoospermia (1.8 %). Conclusions: Abnormal semen parameters remain significant causes in overall infertility in our set up with asthenozoospermia and azoospermia were common abnormalities in male partner. Semen analysis is an inevitable tool for evaluation of infertility in male partner. Further study is required to find out the possible etiologies of male infertility for holistic management of infertility.

Sperm Vitality and Necrozoospermia: Diagnosis, Management, and Results of a Global Survey of Clinical Practice.

Sperm vitality testing is a basic semen examination that has been described in the World Health Organization (WHO) Laboratory Manual for the Examination and Processing of Human Semen from its primary edition, 40 years ago. Several methods can be used to test sperm vitality, such as the eosin-nigrosin (E-N) stain or the hypoosmotic swelling (HOS) test. In the 6th (2021) edition of the WHO Laboratory Manual, sperm vitality assessment is mainly recommended if the total motility is less than 40%. Hence, a motile spermatozoon is considered alive, however, in certain conditions an immotile spermatozoon can also be alive. Therefore, the differentiation between asthenozoospermia (pathological decrease in sperm motility) and necrozoospermia (pathological decrease in sperm vitality) is important in directing further investigation and management of infertile patients. The causes leading to necrozoospermia are diverse and can either be local or general, testicular or extra-testicular. The andrological management of necrozoospermia depends on its etiology. However, there is no standardized treatment available presently and practice varies among clinicians. In this study, we report the results of a global survey to understand current practices regarding the physician order of sperm vitality tests as well as the management practices for necrozoospermia. Laboratory and clinical scenarios are presented to guide the reader in the management of necrozoospermia with the overall objective of establishing a benchmark ranging from the diagnosis of necrozoospermia by sperm vitality testing to its clinical management.

Semen quality and fertility rates in Taipei city

Background: Although female factors are the major focus of attention for infertility, male factors are responsible for approximately 40% of infertility problems. Aims: The aim of this study is to investigate male infertility in Taiwan, establish a database of semen quality, and explore associations between semen quality and infertility. Methods: Results of semen tests of males who were examined at Zhongxiao Branch, Taipei City Hospital, between January 2008 and July 2015 were collected and analyzed retrospectively. All examination procedures and processing of human semen were performed according to the World Health Organization laboratory manual. Total semen volume and the concentration, motility, and morphology of sperm were evaluated and recorded. Pearson's correlation was used to explore associations between age and all semen parameters. Results: Significant differences were found in sperm concentration between the examined years (P = 0.011). Bonferroni post hoc analysis showed that the sperm concentration in 2012 decreased significantly (P = 0.026) compared to data of 2014. Age was significantly associated with sperm motility and morphology (P

Effect of recombinant β-defensin 1 protein on human sperm motility and viability.

The reduction of sperm motility and subsequently reduced ability to undergo capacitation and acrosome reaction are considered as common causes of male infertility. The β-defensin family is a group of well-known secretory proteins with antimicrobial activity that contribute to the process of "sperm maturation" during the passage of spermatozoa in the epididymis when spermatozoa attain its motility. One member of this family is "β-defensin 1" which is present in seminal plasma and spermatozoa. The aim of this study was the incubation of human processed spermatozoa with recombinant β-defensin 1 (500ng/ml) for 1, 2 and 3hr at 37°C under 5% CO2 atmosphere and assessment of sperm viability and motility in 59 semen samples. The analysis of semen samples such as sperm concentration, motility, viability, morphology and semen volume was performed according to the World Health Organization (2010; World health organization laboratory manual for the examination and processing of human semen (p. 287). Geneva, Switzerland: World Health Organization) criteria. The result of the current study shows that the incubation of spermatozoa with recombinant β-defensin significantly maintained percentage of sperm viability and motility compared to processed spermatozoa incubate in the absence of β-defensin in the studied time intervals (p<.05). Therefore, we concluded that recombinant β-defensin 1 protein as an agent with antimicrobial activity can maintain sperm viability and motility in in vitro condition.

Morphological indices for canine spermatozoa based on the World Health Organization laboratory manual for human semen.

Attempting to contribute to the development of a more objective morphological evaluation of dog spermatozoa, in this study the indices of multiple sperm defects (multiple abnormalities index [MAI]; teratozoospermic index [TZI]; sperm deformity index [SDI]) were calculated following the World Health Organization (WHO) guidelines. In Experiment I, the concordance of MAI, TZI and SDI with the proportions of morphologically normal spermatozoa (MNS) was evaluated in fresh ejaculated spermatozoa (dogs=47). In Experiment II, the potential role of indices as prognostic values was assessed in spermatozoa of different origin and treatment (fresh ejaculated: n=6; fresh epididymal: n=6; frozen-thawed ejaculated: n=6) by their correlation with different semen parameters (motility, membrane integrity and acrosome status) and with an in vitro sperm function test. Samples with different proportions of MNS showed different values of SDI, the index that better represented the decline of sperm morphology in both fresh and frozen-thawed samples (Exp. I and II; p<0.05). No correlations between indices and semen parameters were observed (Exp. II), but when samples were evaluated collectively, negative correlations (SDI-motility, p=0.01; SDI-acrosome integrity, p=0.002) were found. Including all the defects of each spermatozoon, SDI might be a useful index during morphological analysis and better discriminates the increase in multiple defects. A more objective morphological evaluation for dog spermatozoa was achieved by the WHO method, and in vitro tests allowed to elucidate the validity of SDI as prognostic indicator of in vitro fertilizing potential.

Decline in semen quality in men in northern Taiwan between 2001 and 2010.

Semen quality is a key indicator of the male reproductive ability. The decline in semen quality has been debated for several decades. However, data on decline in semen quality might be inconsistent given factors such as geographical area, ethnicity, environmental exposure and lifestyle changes. This study aimed to provide information on the semen quality of men in Northern Taiwanese. We recruited 7,187 male participants between 2001 and 2010 from a reproductive medical center. The age of the participants ranged from 26 to 57 years, with a mean age of 36.9 ± 4.77 years. Semen analysis was performed through standardized methods outlined in the World Health Organization laboratory manual. Increasing age (per year) was significantly and negatively associated with semen volume (1.006 ml), progressive sperm motility, rapid progressive sperm motility and sperm with normal morphology (reduction by 1.010%, 1.013% and 1.002% per year, respectively). In addition, sperm concentration, semen volume, number of sperms, progressive sperm motility, rapid progressive sperm motility, and sperm with normal morphology were significantly reduced annually by 1.013 × 10⁶/ml, 1.015 ml, 1.028 × 10⁶, 1.021 %, 1.017% and 1.016%, respectively). Age-period-cohort (APC) analyses revealed that age and cohort had effects on reduction of the progressive and rapid progressive sperm motility. Moreover, all the sperm parameter values were significantly reduced annually. Our findings provide useful information for clinical practice and public health investigations of male reproductive health.

Uncertainty associated with assessing semen volume: are volumetric and gravimetric methods that different?

The World Health Organization laboratory manual for the examination of human semen suggests that an indirect measurement of semen volume by weighing (gravimetric method) is more accurate than a direct measure using a serological pipette. A series of experiments were performed to determine the level of discrepancy between the two methods using pipettes and a balance which had been calibrated to a traceable standard. The median weights of 1.0ml and 5.0ml of semen were 1.03 g (range 1.02–1.05 g) and 5.11 g (range 4.95–5.16 g), respectively, suggesting a density for semen between 1.03g and 1.04 g/ml. When the containers were re-weighed after the removal of 5.0 ml semen using a serological pipette, the mean residual loss was 0.12 ml (120 μl) or 0.12 g (median 100 μl, range 70–300 μl). Direct comparison of the volumetric and gravimetric methods in a total of 40 samples showed a mean difference of 0.25ml (median 0.32 ± 0.67ml) representing an error of 8.5%. Residual semen left in the container by weight was on average 0.11 g (median 0.10 g, range 0.05–0.19 g). Assuming a density of 1 g/ml then the average error between volumetric and gravimetric methods was approximately 8% (p < 0.001). If, however, the WHO value for density is assumed (1.04 g/ml) then the difference is reduced to 4.2%. At least 2.4–3.5% of this difference is also explained by the residual semen remaining in the container. This study suggests that by assuming the density of semen as 1 g/ml, there is significant uncertainty associated with the average gravimetric measurement of semen volume. Laboratories may therefore prefer to provide in-house quality assurance data in order to be satisfied that ‘estimating’ semen volume is ‘fit for purpose’ as opposed to assuming a lower uncertainty associated with the WHO recommended method.

Effect of varicocele on semen characteristics according to the new 2010 World Health Organization criteria: a systematic review and meta-analysis.

This study investigated the effects of varicocele on semen parameters in infertile men based on the new 2010 World Health Organization laboratory manual for the examination of human semen. Semen analysis results (volume, sperm count, motility, and morphology) were the primary outcomes. An electronic search to collect the data was conducted using the Medline/PubMed, SJU discover, and Google Scholar databases. We searched articles published from 2010 to August 2015, i.e., after the publication of the 2010 WHO manual. We included only those studies that reported the actual semen parameters of adult infertile men diagnosed with clinical varicocele and contained a control group of either fertile men or normozoospermic men who were not diagnosed with varicocele. Ten studies were included in the meta-analysis, involving 1232 men. Varicocele was associated with reduced sperm count (mean difference: −44.48 × 106 ml−1; 95% CI: −61.45, −27.51 × 106 ml−1; P < 0.001), motility (mean difference: −26.67%; 95% CI: −34.27, −19.08; P < 0.001), and morphology (mean difference: −19.68%; 95% CI: −29.28, −10.07; P < 0.001) but not semen volume (mean difference: −0.23 ml; 95% CI: −0.64, 0.17). Subgroup analyses indicated that the magnitude of effect was influenced by control subtype but not WHO laboratory manual edition used for semen assessment. We conclude that varicocele is a significant risk factor that negatively affects semen quality, but the observed pooled effect size on semen parameters does not seem to be affected by the WHO laboratory manual edition. Given most of the studies published after 2010 still utilized the 1999 manual for semen analysis, further research is required to fully understand the clinical implication of the 2010 WHO laboratory manual on the association between varicocele and semen parameters.