Conventional Semen Parameters Research Articles

Are oxidative stress markers associated with unexplained male infertility?

Male infertility can be responsible for up to 20% of the cases attending fertility consultation facilities; nonetheless, the underlying molecular mechanisms that could explain it are still elusive. Therefore, we aimed to evaluate conventional and functional parameters of semen samples from patients who presented with male infertility of unknown origin. Conventional semen parameters and functional parameters (i.e. intracellular reactive oxygen species production, mitochondrial membrane potential, sperm chromatin structure assay, sperm membrane lipid peroxidation and antioxidant capacity of seminal plasma) were evaluated on semen samples from 54 healthy donors, 23 patients with idiopathic infertility and 34 fertile controls. No significant differences were observed in the conventional seminal parameters between the fertile and infertile men. However, increased intracellular reactive oxygen species (ROS) production and DNA fragmentation were observed in the infertile patients compared to the fertile group. Alterations in intracellular ROS production and DNA fragmentation could be associated with male idiopathic infertility. These parameters could eventually distinguish both groups more accurately than the conventional parameters. Our current results are encouraging, and the efficacy of these parameters in the clinical settings needs to be further assessed to establish their predictive potential as a marker of unexplained male infertility.

Should we evaluate and treat sperm DNA fragmentation?

The clinical utility of sperm DNA fragmentation tests needs to be revisited in light of increasing evidence of detrimental effect of sperm DNA damage on reproductive outcomes. Current evidence supports the association between high sperm DNA fragmentation and poor outcomes with regards to natural conception and intrauterine insemination. The relationship between high sperm DNA fragmentation and impaired outcomes after in-vitro fertilization and intracytoplasmic sperm injection are more equivocal. However, recent studies indicate that poor sperm chromatin content is associated with an increased risk of early pregnancy loss after in-vitro fertilization and intracytoplasmic sperm injection. Several strategies are proposed to alleviate sperm DNA fragmentation and/or select sperm with higher quality chromatin content for assisted reproductive techniques. The intake of oral antioxidants, varicocele repair, use of recurrent ejaculations alone or combined with micromanipulation-based sperm selection techniques, and the use of testicular sperm for intracytoplasmic sperm injection have been attempted with promising results. Sperm DNA fragmentation tests provide clinically relevant information for natural conception and artificial reproduction independent of those derived from conventional semen parameters. The increasing knowledge of paternal factors on pregnancy outcome and the improvement in treatment strategies should prompt routine evaluation of sperm DNA fragmentation in infertile couples.

Effects of n-6 and n-3 polyunsaturated acid-rich soybean phosphatidylcholine on membrane lipid profile and cryotolerance of human sperm

To study the effects of n-6 and n-3 polyunsaturated acid-rich soybean phosphatidylcholine (soy-PC) on sperm cryotolerance with regard to sperm membrane lipid profile, membrane surface integrity, and routine semen parameters. Experimental study. University-affiliated tertiary hospital. A total of 20 normospermic fertile men. Semen samples examined for differences in semen parameters, sperm membrane lipid profile, and plasma membrane surface both before and after cryopreservation using basic freezing medium with N-tris(hydroxymethyl)-methyl-2-aminoethane sulfonic acid (TES) and tris-(hydroxymethyl)-aminomethane (TRIS) supplemented with purified soy-PC (TEST-PC) or egg yolk (TEST-Y), both alone or in association (TEST-Y-PC). Conventional semen parameters and membrane lipid profile by matrix-assisted laser/desorption ionization mass spectrometry (MALDI-MS). Postthaw sperm cell motility, vitality, and morphology parameters were similar for soy-PC (TEST-PC) and egg yolk (TEST-Y) cryoprotectants. However, sperm exposed to TEST-Y-PC presented better kinetic parameters, which were similar to the original quality of the fresh semen. Human sperm MALDI-MS lipid profiles revealed that the relative abundance of glycerophospholipids of m/z 760.44 [PC (34:1)+H]+, 781.55 [SM (20:0) +Na]+, 784.55 [PC (36:3) +H]+, 806.64 [PC (38:6) +H]+, 807.64 [SM (22:1) +Na]+, and 809.64 [SM (22:0) +Na]+ increased in soy-PC samples (TEST-PC). Nonetheless, only one lipid (m/z 781.55, [SM (20:0) +Na]+) statistically significantly changed when sperm was cryopreserved in TEST-Y-PC. Sphingomyelin was defined as a prospective biomarker of soy-PC treatment, and it could be related to the positive cryoprotective effects of soy-PC in human sperm, opening new perspectives to design of a more efficient synthetic cryoprotectant medium containing purified egg yolk biomolecules combined with soy-PC.

Comparison of Conventional Semen Parameters and Hypo-osmotic Swelling Test between Karan Fries and Sahiwal Bulls under Heat Stress

The present study aimed to determine the differences in semen parameters between Karan Fries (KF) and Sahiwal bulls and the correlation of hypo-osmotic swelling test with conventional semen parameters. A total of 24 ejaculates were collected from each breed and subsequently classified according to their mass activity. The semen samples were further examined for progressive motility, sperm viability, sperm concentration and hypo-osmotic swelling test (HOST). The percentage of hypo-osmotically swelled spermatozoa was higher (P 0.05) between the breeds. With respect to association between HOST and conventional semen parameters, HOST showed significant (P<0.01) positive correlation with progressive motility (%), sperm viability (%) and sperm concentration in both breeds. This study indicated the HOST as an effective method to test the fertility potential of semen samples for artificial insemination and zebu breeds ejaculates have more fertility potential as compared to crossbred ejaculates under heat stress.

Summary evidence on the effects of varicocele treatment to improve natural fertility in subfertile men.

The objective of this review was to summarize the evidence concerning the benefit of varicocele treatment to improve natural fertility in subfertile males. We also analyzed the effect of varicocele treatment on conventional semen parameters and sperm functional tests. An electronic search to collect the data was performed using the PubMed/MEDLINE databases until July 2015. Data pooled from a variety of study designs indicate that varicocelectomy improves semen parameters in the majority of the treated men with clinical varicocele and abnormal semen parameters regardless of the chosen surgical method. Surgical varicocele repair was beneficial not only for alleviating oxidative stress-associated infertility but also to improve sperm nuclear DNA integrity. However, given the low magnitude of the effect size in sperm DNA integrity, further research is needed to elucidate its clinical significance. Conflicting results on the effect of varicocele treatment on natural fertility seem to be due to heterogeneous study designs and, more importantly, patient selection criteria. When these issues are controlled, current evidence indicates that treatment of subclinical varicocele is not warranted, as it does not seem to improve fertility. On the contrary, fair evidence indicates that varicocele treatment should be offered to infertile patients with palpable varicocele and abnormal semen parameters. This evidence supports the current guidelines issued by the American Urological Association and European Association of Urology, which state that varicocele treatment should be offered to male partners of infertile couples presenting for evaluation with clinical varicocele and semen parameters alterations.

Body mass index and human sperm quality: neither one extreme nor the other

The aim of the present study was to investigate the still contentious association between body mass index (BMI) and seminal quality. To this end, 4860 male patients (aged 18-65 years; non-smokers and non-drinkers), were classified according to BMI as either underweight (UW; BMI <20kgm-2; n=45), normal weight (NW; BMI 20-24.9kgm-2; n=1330), overweight (OW; BMI 25-29.9kgm-2; n=2493), obese (OB; BMI 30-39.9kgm-2; n=926) or morbidly obese (MOB; BMI ≥40kgm-2; n=57). Conventional semen parameters and seminal concentrations of fructose, citric acid and neutral α-glucosidase (NAG) were evaluated. The four parameters that reflect epididymal maturation were significantly lower in the UW and MOB groups compared with NW, OW and OB groups: sperm concentration, total sperm count (103.3±11.4 and 121.5±20.6 and vs 157.9±3.6, 152.4±2.7 or 142.1±4.3 spermatozoa ejaculate-1 respectively, P<0.05), motility (41.8±2.5 and 42.6±2.6 vs 47.8±0.5, 48.0±0.4 or 46.3±0.6 % of motile spermatozoa respectively, P<0.05) and NAG (45.2±6.6 and 60.1±7.9 vs 71.5±1.9, 64.7±1.3 or 63.1±2.1 mU ejaculate-1 respectively, P<0.05). Moreover, the percentage of morphologically normal spermatozoa was decreased in the MOB group compared with the UW, NW, OW and OB groups (4.8±0.6% vs 6.0±0.8%, 6.9±0.1%, 6.8±0.1 and 6.4±0.2%, respectively; P<0.05). In addition, men in the MOB group had an increased risk (2.3- to 4.9-fold greater) of suffering oligospermia and teratospermia (P<0.05). Both morbid obesity and being underweight have a negative effect on sperm quality, particularly epididymal maturation. These results show the importance of an adequate or normal bodyweight as the natural best option for fertility, with both extremes of the BMI scale as negative prognostic factors.

Hyaluronic Acid Binding Assay Is Highly Sensitive to Select Human Spermatozoa with Good Progressive Motility, Morphology, and Nuclear Maturity

Objective: This study was performed to evaluate the correlation of hyaluronic acid binding assay (HBA) with conventional semen parameters, lipid peroxidation (LPO), intracellular reactive oxygen species (ROS), DNA fragmentation (DF), DNA maturity and mitochondrial membrane potential (MMP) level in human spermatozoa. Methods: The semen samples were obtained from 98 patients. The seminal plasma was separated for the study of LPO, and the pellet was employed for evaluation of intracellular ROS, DF, nuclear maturity (sperm chromatin structure assay) and MMP through flowcytometry. Results: The correlation and strength of HBA with respect to the studied parameters were estimated by the Pearson coefficient and multiple liner regression tests. While HBA indicated a positive correlation with progressive motility (β-coefficients = 0.449, p < 0.05) and normal morphology (β-coefficients = 2.722, p < 0.01), it had only negative relationship with DNA integrity (high DNA stain ability; β-coefficients = -0.517, p < 0.05). HBA also did not show any important correlation with other conventional and intracellular sperm parameters. Conclusions: The HBA is sensitive to morphological integrity, high progressive motility and nuclear maturation. Nonetheless, HBA is not a reliable test for prediction of sperm intracellular ROS, DF and MMP risks and healthy spermatozoa selection.

Analysis of the correlation between sperm DNA integrity and conventional semen parameters in infertile men

A male factor is responsible in approximately 30-40% of couples receiving infertility treatment. Routinely, such couples undergo semen analysis including parameters such as sperm count, motility and morphology. Generally, the analysis of sperm DNA damage, shown to have a significant clinical importance by many studies, is recognized as an advanced test that is not included in routine infertility tests. Intracytoplasmic sperm injection method, commonly employed in the current infertility treatment protocols, lowers the fertilization rate, however, fertilization can occur even with a damaged DNA which is known to pose a risk in the subsequent pregnancy period. The relation between sperm morphology and the degree of sperm DNA damage has not yet been understood clearly. In this study, we aimed to investigate the association between routine semen analysis and sperm DNA integrity assay, another advanced but costly method. The degree of DNA damage was compared with the results of semen analysis, based on the WHO criteria, in 399 male patients who received comet assay for sperm DNA integrity. The statistical correlation analyses were performed with Windows SPPS statistical package program. Accordingly, the sperm DNA damage was found to be correlated with all 3 parameters (sperm count, forward motility, and morphology) examined by the semen analysis (p<0.001). Total sperm DNA Damage Count was 226, 216, and 210 arbitrary units in patients with a sperm count <15 mil/mL, forward moving motility <32%, and normal morphology <4%, respectively. The difference with the normal individuals was statistically significant (p<0.001). In light of the comet assay results, higher degree of sperm DNA damage is associated with significant impairment of all seminal parameters.

Evaluating γH2AX in spermatozoa from male infertility patients

To investigate whether γH2AX levels were different in the spermatozoa of healthy men compared with infertility patients, and to assess the possible correlations between γH2AX and conventional semen parameters and double-stranded breaks (DSBs) identified with the use of comet assay. Prospective study. Clinical laboratory. Semen from 100 male infertile patients and 100 healthy sperm donors. Human sperm samples were analyzed in terms of World Health Organization parameters. The γH2AX levels were detected by means of flow cytometry. DSBs of sperm were detected by means of comet assay. Morphology slides were made and the sperm morphology assessed according to strict criteria. Conventional semen analyses, γH2AX levels in sperm, DNA DSBs in sperm, and correlations among γH2AX, conventional semen analyses, and DSBs. Concentration, viability, motility, and normal sperm morphology were significantly lower in male infertility patients compared with healthy men. Also, γH2AX levels and the number of DSBs were significantly higher in the sperm of infertile subjects compared with healthy men. γH2AX levels correlated negatively with conventional semen parameters and positively with DSBs. A threshold γH2AX level of 18.55% was identified as a cutoff value to discriminate infertile subjects from fertile control subjects with a specificity of 86.0% and a sensitivity of 83.0%. The positive and negative predictive values of the 18.55% γH2AX threshold were high: 87.7% and 85.5%, respectively. γH2AX levels were higher in the sperm of male infertility patients than in healthy men. γH2AX levels in sperm, as evaluated with the use of flow cytometry, might be a useful biomarker for evaluating DSBs in human spermatozoa.

Two-dimensional polyacrylamide gel electrophoresis of Bali bull (Bos javanicus) seminal plasma proteins and their relationship with semen quality

The present study evaluated the relationship between Bali bull (Bos javanicus) seminal plasma proteins and different semen quality parameters. Semen samples from 10 mature Bali bulls were evaluated for conventional semen parameters (general motility, viability, and normal morphology), sperm functionality (acrosome reaction, sperm penetration rate, sperm penetration index), sperm kinetics (computer-assisted semen analysis parameters such as sperm velocity), and sperm morphology (acrosome and membrane integrity). Frozen–thawed semen with higher sperm motility, viability, acrosome integrity, and membrane integrity (P < 0.05) are consistently higher in acrosome reaction and sperm penetration assay. Three bulls showed the highest, four bulls displayed the medium, and the remaining three bulls showed the lowest for all sperm parameters and SPA. The proteome maps of seminal plasma from high-quality and low-quality Bali bulls were also established. Seminal plasma of both high-quality and low-quality Bali bulls was subjected to two-dimensional SDS-PAGE with isoelectric point ranged from 3 to 10 and molecular weight from 10 to 250 kDa. Approximately 116 spots were detected with Blue Silver stain, and of these spots, 29 were selected and identified by MALDI-TOF/TOF-MS/MS. A majority of the proteins visualized in the seminal plasma two-dimensional maps was successfully identified. An essential group of the identified spots represented binder of sperm 1 (BSP1), clusterin, spermadhesin, tissue inhibitor of metalloproteinases 2 (TIMP-2), and phospholipase A2 (PLA2). Other proteins found in high abundance included seminal ribonuclease, serum albumin, cationic trypsin, and peptide similar to β2 microglobulin. Thus, a reference map of Bali bull seminal plasma proteins has been generated for the very first time and can be used to relate protein pattern changes to physiopathologic events that may influence Bali bull reproductive performance.

Conventional semen parameters and DNA fragmentation in relation to fertility status in a Greek population

ObjectivesOur study aimed to investigate the possible correlations between conventional seminal parameters and DNA fragmentation in specific groups of Greek men, selected in relation to their fertility history and to verify the validity of the recent WHO reference values for the basic semen analysis in this population. Study designA total of 770 subject data were evaluated in three distinct groups: fertile men with children naturally conceived within one year of unprotected intercourse (n=78), subfertile men, having achieved pregnancies either naturally or by Assisted Reproduction Techniques, not resulting in live births (n=153) and infertile men, failing to produce either pregnancies or children (n=539). Semen volume, sperm concentration, total count, rapid and total progressive motility and morphology were evaluated following the World Health Organization (2010) methods. DNA fragmentation was assessed by the Sperm Chromatin Dispersion assay. The 5th percentile, as well as the 95% confidence intervals (CI) for each parameter, were calculated by the method of bootstrapping. Statistical correlations between the examined parameters were sought using the Spearman R test (p<0.05). ResultsA significant inverse correlation was established between DNA fragmentation and sperm concentration, total count, progressive motility (rapid and total) and normal morphology in subfertile and infertile men (p<0.05). No statistically significant correlations were remarked between the conventional semen parameters and the levels of DNA fragmentation in our group of fertile men. Concordance was established between the reference limits issued by the WHO 2010 for the basic semen parameters and semen quality of fertile men in the studied population. ConclusionsThe variability of correlations established between DNA fragmentation and the conventional seminal profile in relation to fertility status indicates that they are independent attributes of semen quality, justifying the assessment of both during a comprehensive evaluation of male infertility. Moreover, the WHO 2010 reference limits were found adequately descriptive of seminal normality in Greek men.

Are semen quality parameters sufficient for biomonitoring spermatozoa DNA integrity and oxidatively damaged DNA

Abstract:The present study aimed to investigate the relationship between semen quality parameters and DNA integrity, and determine whether semen quality parameters could serve as a reliable biomarker for monitoring sperm DNA damage. Conventional semen parameters from a total of 202 male human subjects were analyzed. DNA fragmentation and 8-oxo-7,8-dihydro-2′- deoxyguanosine (8-oxoGuo) were used to assess sperm DNA integrity. DNA fragmentation was analyzed by the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay and sperm chromatin structure assay (SCSA), while 8-oxodGuo was quantified by the liquid chromatography/tandem mass spectrometry (LC-MS/MS) coupled with an on-line solid phase system. The levels of 8-oxodGuo levels in sperm were related to the percentages of DNA fragmentation measured by both the TUNEL and SCSA (r = 0.22, p = 0.048; r = 0.12, p = 0.039). Sperm vitality, motility and morphology from all of the participants exhibited a weak correlation with the levels of 8-oxodGuo and the percentages of DNA fragmentation. Semen quality parameters may be independent of the formation of DNA fragmentation and oxidative adducts in sperm. Semen quality parameters may be insufficient to monitor sperm DNA fragmentation and oxidative damage. DNA damage in sperm is recommended to be included in routine measurements.

Effect of Body mass index on interleukin2, 6 and soluble fibroblast associated surface antigen in infertile men

Background: obesity is an important cause of adverse health problems, including male infertility. Testosterone is essential for spermatogenesis and permits the release of mature sperms. In the absence of testosterone stimulation, spermatogenesis does not proceed beyond the meiosis stage. Aromatase expression is directly related to the degree of adiposity; it is dependent on cytokine stimulation and requires the presence of glucocorticoids, 17β-estradiol in the plasma of adult men is formed by aromatization convert testosterone and androstinedione to 17β-estradiol Apoptosis is an important process in the context of germ cells since they undergo both mitosis and meiosis, and this process is affected by interleukin 6and 2 (IL6 and IL2).Objectives: To assess the effect of body mass index on serum sex steroidal hormones, seminal antiapoptotic factor soluble fibroblast associated surface antigen (sFas) and inflammatory markers (IL6, IL-2), with conventional semen parameters in infertile men.Patients and methods: One hundred and six male partners of infertile couples were involved in the study. Height and weight of them were measured to calculate the body mass index (BMI). Serum testosterone and estradiol 2.were measured for all.Semen sample was taken after 2-7 days of abstinence. Conventional semen analysis was done for each sample according to the protocol of (WHO) 2010, after incubation and liquefaction period (30-60 min). Semen plasma was collected for analysis of interleukins (2 and 6) and sFAS .by specific kits Patients with severe oligospermia (below million sperm/ml) were excluded from the study. The rest (77) subjects were divided into three groups according to BMI (WHO, 2010) as follows: Normal (BMI &lt; 25 kg/ m2) (n=24): Overweight (BMI ≥ 25 and &lt; 30 kg/m2) (n=30): Obese (BMI &gt; 30 kg/m2) (n=23).Results: The results showed a significant decrease in Testosterone level and Testosterone/E2 ratio in overweigh and obese subjects as compared to normal. The results of conventional semen analysis showed significant difference in pH and no. of round cell between the two groups. There was positive correlation between IL2 with sFAS in the two groups, and negative correlation of IL2 with total progressive cell and motility in both overweigh and obese groups. There was positive correlation of IL6 with no. of round cells in both studied groups.Conclusion: BMI affects Testosterone level and Testosterone/E2 ratio (they were significantly decreased in overweigh and obese patients). A positive correlation between IL2 and sFAS in obese patients as well as positive correlation of IL6 with round cells in obese patients indicate the role of apoptosis which is essential for normal fertility to remove abnormal and unwanted cells in ejaculate, in infertile overweight and obese patients.

Negative biomarker based male fertility evaluation: Sperm phenotypes associated with molecular-level anomalies.

Biomarker-based sperm analysis elevates the treatment of human infertility and ameliorates reproductive performance in livestock. The negative biomarker-based approach focuses on proteins and ligands unique to defective spermatozoa, regardless of their morphological phenotype, lending itself to analysis by flow cytometry (FC). A prime example is the spermatid specific thioredoxin SPTRX3/TXNDC8, retained in the nuclear vacuoles and superfluous cytoplasm of defective human spermatozoa. Infertile couples with high semen SPTRX3 are less likely to conceive by assisted reproductive therapies (ART) and more prone to recurrent miscarriage while low SPTRX3 has been associated with multiple ART births. Ubiquitin, a small, proteolysis-promoting covalent posttranslational protein modifier is found on the surface of defective posttesticular spermatozoa and in the damaged protein aggregates, the aggresomes of spermiogenic origin. Semen ubiquitin content correlates negatively with fertility and conventional semen parameters, and with sperm binding of lectins LCA (Lens culinaris agglutinin; reveals altered sperm surface) and PNA (Arachis hypogaea/peanut agglutinin; reveals acrosomal malformation or damage). The Postacrosomal Sheath WWI Domain Binding Protein (PAWP), implicated in oocyte activation during fertilization, is ectopic or absent from defective human and animal spermatozoa. Consequently, FC-parameters of PAWP correlate with ART outcomes in infertile couples and with fertility in bulls. Assays based on the above biomarkers have been combined into multiplex FC semen screening protocols, and the surface expression of lectins and ubiquitin has been utilized to develop nanoparticle-based bull semen purification method validated by field artificial insemination trials. These advances go hand-in-hand with the innovation of FC-technology and genomics/proteomics-based biomarker discovery.

Infertile men older than 40years are at higher risk of sperm DNA damage.

BackgroundThe effect of paternal age on semen quality is controversial. In this retrospective study, the aim was to investigate the effects of advancing age on sperm parameters including reactive oxygen species (ROS), total antioxidant capacity (TAC) and sperm DNA damage in infertile men. We also examined whether paternal age >40 y is associated with higher risk of sperm DNA damage.MethodsA total of 472 infertile men presenting for infertility were divided into 4 age groups: group A: patients ≤ 30 y; group B: patients 31- 40 y, group C: ≤ 40 y and group D: patients >40 y. The following tests were performed - semen analysis according to WHO 2010 criteria, seminal ROS by chemiluminescence, TAC by colorimetric assay and sperm DNA damage by TUNEL assay - and the results were compared amongst the 4 age groups.ResultsThere was no statistical difference in conventional semen parameters, TAC and ROS with advancing paternal age as well as between different age groups. However, a significant negative association was noted between sperm DNA damage and advancing paternal age. Men >40 y showed higher levels of sperm DNA damage (24.4 ± 18.5%) compared to younger men (<30 y; 16.7 ± 11.2%; p <0.05).ConclusionsInfertile men over the age of 40 y have a greater percentage of sperm DNA fragmentation compared to infertile men aged 40 y and below. Advanced paternal age (>40 y) may increase the risk of sperm DNA damage in infertile men.

The Correlation between Ultrasound Testicular Volume and Conventional Semen Parameters in Albanian Subfertile Males

OBJECTIVE: The study is conducted to evaluate the relationship between testicular volume measured by ultrasound and conventional sperm parameters (volume, concentration, total count, motility and morphology) in Albanian subfertile males and to determine a normal limit value of the testicular volume.MATERIALS AND METHODS: A total of 500 males were observed for this study. The testicular volumes of all subjects were measured by ultrasonography. The semen samples were collected by the process of masturbation after 3-5 days of ejaculatory abstinence and were analyzed according to WHO criteria 2010.RESULTS: Testicular volume has a strong positive correlation with sperm count, total count and motility and a positive correlation with semen volume.CONCLUSION: Testicular volume has a direct correlation with semen parameters and the critical total testicular volume indicating normal testicular function is approximately 26.6 ml (the mean testicular volume 13.3 ml). The measurement of testicular volume can be helpful for assessing fertility at the initial physical examination.

Relationship between human spermatozoa-hyaluronan-binding assay, conventional semen parameters and fertilisation rates in intracytoplasmic spermatozoa injection.

Selecting the best spermatozoa for intracytoplasmic spermatozoa injection (ICSI) has recently been a topic of great interest among embryologists. The study aimed to evaluate the relationship between the spermatozoa-hyaluronan-binding assay (HBA), routine semen analysis results and fertilisation rates as recorded during conventional ICSI therapy. Ninety-one patients undergoing conventional ICSI treatment in the Medfem Fertility Clinic in Johannesburg (South Africa) were included in the study. A total of 797 oocytes were injected of which 457 oocytes fertilised (57.3%, range 0-100%). None of the semen parameters correlated with the fertilisation rates (Table 2). HBA scores, however, revealed a highly significant association (p ≤ 0.0001) with the fertilisation rates. The HBA scores also correlated significantly with the biochemical pregnancy values (Spearman r = 0.24, P = 0.02, 95% CI 0.039-0.43); however, the HBA scores did not correlate with the clinical pregnancy rates (Spearman r = 0.14, P = 0.16, 95% CI -0.06 to 0.34). No correlation was recorded between HBA and the standard semen parameters. The study showed that HBA is significantly associated with fertilisation in conventional ICSI. The HBA scores were also significantly associated with the fertilisation rates and biochemical pregnancies.

Sperm vacuoles cannot help to differentiate fertile men from infertile men with normal sperm parameter values.

Can the assessment of sperm vacuoles at high magnification contribute to the explanation of idiopathic infertility? The characteristics of sperm head vacuoles (number, area, position) are no different between fertile controls and patients with unexplained infertility. Until now, the assessment of sperm head vacuoles has been focused on a therapeutic goal in the intracytoplasmic morphologically selected sperm injection (IMSI) procedure, but it could be pertinent as a new diagnostic tool for the evaluation of male fertility. This diagnostic test study with blind assessment included a population of 50 fertile men and 51 men with idiopathic infertility. They were selected from September 2011 to May 2013. Fertile men were within couples who had a spontaneous pregnancy in the last 2 years. Infertile men were within couples who had unexplained infertility and were consulting in our centre. After analysis of conventional sperm parameters, we investigated the number, position and area of sperm head vacuoles at high magnification (×6000) with interference contrast using an image analysis software. We also carried out a nuclear status analysis by terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling assay (TUNEL), sperm chromatin structure assay (SCSA) and aniline blue staining. Concerning the vacuoles data, we did not find any significant difference between the two populations. We found no significant correlation between the vacuolar parameters (mean number of vacuoles, relative vacuole area and percentage of spermatozoa with large vacuoles) and either conventional semen parameters, male age or the data from the aniline blue staining, SCSA assay and TUNEL assay. Despite the fact all of the vacuole parameters values were identical in fertile and infertile men, we cannot totally exclude that a very small cause of unexplained infertilities could be related to an excess of sperm vacuoles. In line with its widely debated use as a therapeutic tool, sperm vacuole assessment for diagnostic purposes does not seem useful. The study was funded by a grant from Association pour la Recherche sur les Traitements de la Stérilité. There are no competing interests to declare.

The Correlation between Ultrasound Testicular Volume and Conventional Semen Parameters in Albanian Subfertile Males

OBJECTIVE: The study is conducted to evaluate the relationship between testicular volume measured by ultrasound and conventional sperm parameters (volume, concentration, total count, motility and morphology) in Albanian subfertile males and to determine a normal limit value of the testicular volume.MATERIALS AND METHODS: A total of 500 males were observed for this study. The testicular volumes of all subjects were measured by ultrasonography. The semen samples were collected by the process of masturbation after 3-5 days of ejaculatory abstinence and were analyzed according to WHO criteria 2010.RESULTS: Testicular volume has a strong positive correlation with sperm count, total count and motility and a positive correlation with semen volume.CONCLUSION: Testicular volume has a direct correlation with semen parameters and the critical total testicular volume indicating normal testicular function is approximately 26.6 ml (the mean testicular volume 13.3 ml). The measurement of testicular volume can be helpful for assessing fertility at the initial physical examination.

Effect of advancing paternal age on semen parameters and seminal oxidative stress markers in infertile men

BackgroundHigh rate of subfertility and adverse pregnancy related out-comes associated with childbearing are seen after age 40.In contrast to oogenesis, spermatogenesis continues inelderly men. Adult male germ cells pass through signifi-cantly more mitotic replications than the germ cells inadult female. In men, there is an age associated increase inthe incidence of breaks in sperm DNA, decrease in apopto-sis, and a higher frequency of point mutations. Advancedpaternal age is associated with an increased time to preg-nancy and decreased pregnancy rates. After adjusting forfemale age, conception during a 12-month period was > 30percent less likely for men over 40 years of age as com-pared to men 45years compared to men 40 years (n = 105). We evaluated the conven-tional semen parameters (WHO, 2010) [5] and OS mar-kers: seminal ROS (chemiluminescence assay), totalantioxidant capacity (TAC) and sperm DNA damage(TUNEL assay) in all patients.ResultsThe mean age of the study subjects was 36.8 ± 6.7 years.No age-related differences were seen in conventionalsemen parameters (volume, concentration, motility, andmorphology) (Table 1). ROS and antioxidant levels werecomparable in the 3 groups. Significantly higher levels ofsperm DNA damage (19.94 ± 15.30%) was seen in infertilemen >40 years compared to men in younger age groups(P = 0.028 and P = 0.027, respectively).ConclusionsSperm DNA damage increases with advancing paternalage. Evaluation of sperm DNA damage will help diagnosethe underlying cause of poor fertility in some men andassist the clinician in offering correct treatment modality.