Assessment Of Sperm Morphology Research Articles

Effects of clinical stage and immunological status on semen analysis results in human immunodeficiency virus type 1-seropositive men.

Complete semen analyses including computer-assisted sperm motility and morphology assessments were performed to determine if semen and sperm differed between HIV-seropositive men and fertile controls, or differed with symptoms, or CD4+ peripheral cell count categories. Previous studies included small numbers of men and presented conflicting conclusions. Two hundred and fifty non-vasectomized HIV-seropositive men and 38 fertile controls each provided one semen sample. Non-parameteric statistics were used to analyse both continuous and nominal data. Fertile men had significantly greater semen volume, sperm concentration, percent motility, percent rapid and linear motility and total strictly normal spermatozoa than HIV seropositive men. Neither total number nor subtypes of leukocytes in semen differed between the two groups. Among the HIV seropositive men, significant differences in semen analyses were found between CD4+ cell count, clinical, and AIDS categories. Lower CD4+ cell counts (< 200 mm-3) were associated with significantly lower percent motility, percent normal sperm morphology by strict criteria, significantly more spermatids in semen, and higher percentages of teratozoospermia, oligoasthenoteratozoospermia and leukocytospermia. Healthier men, based on clinical categories, had significantly more normal shaped spermatozoa and fewer had azoospermia, oligoasthenoteratozoospermia or leukocytospermia. Many HIV-seropositive men have normal semen analyses, but as the disease progresses more defects are found, particularly in strict criteria sperm morphology.

DNA fragmentation of normal spermatozoa negatively impacts embryo quality and intracytoplasmic sperm injection outcome

To evaluate DNA fragmentation in morphologically normal sperm recovered from the same sample used for intracytoplasmic sperm injection (ICSI) and to correlate DNA damage with embryo quality and pregnancy outcome. Prospective study. Academic center. 36 infertile men participating in the ICSI program. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-fluorescein nick end labeling (TUNEL) assay and morphologic assessment by phase contrast. Simultaneous assessment of sperm morphology and DNA fragmentation by TUNEL assay was performed in the same cell, then the percentage of normal sperm with fragmented DNA (normal SFD) was correlated with embryo quality and pregnancy outcomes. A highly statistically significant negative correlation was found between the percentage of normal SFD and embryo quality. This association was confirmed for the transferred embryos and for the total embryo cohort. The receiver operating characteristics curve analysis demonstrated that the percentage of normal SFD and embryo quality were statistically significant predictors of pregnancy. When the percentage of normal SFD was <or=17.6 %, the likelihood of pregnancy was 3.5 times higher. No correlation was found between the percentage of total sperm with fragmented DNA (morphologically normal and abnormal) and ICSI outcomes. The DNA fragmentation of morphologically normal sperm negatively impacts embryo quality and probability of pregnancy in ICSI cycles.

Sperm morphology assessment according to WHO and strict criteria: method comparison and intra-laboratory variability

Sperm morphology assessment according to WHO and strict criteria: method comparison and intra-laboratory variability

Effects of cryopreservation on head morphometry and its relation with chromatin status in brown bear ( Ursus arctos) spermatozoa

The Cantabrian brown bear ( Ursus arctos) is a highly endangered species in Spain and basic studies are necessary in order to bank its germplasm. Sperm heads are mainly made up of chromatin, thus their shape depends partly on chromatin structure. Thawed semen from 10 bears was used to analyze chromatin status by sperm chromatin structure assay (SCSA) and head morphometry by the computer-assisted sperm morphology assessment (CASMA) system. Morphometry was analyzed before and after freezing–thawing in order to evaluate the effects of cryopreservation on sperm heads. Each spermatozoon was measured for four primary parameters (length, L; width, W; area, A; perimeter, P) and derived parameters (ellipticity: L/ W, circularity: 4 πA/ P 2, elongation: ( L − W)/( L + W), regularity: πLW/4 A). All the derived parameters significantly differed between bears. Likewise, cryopreservation affected head morphometry by reducing its size. Clustering based on morphometric parameters separated three subpopulations, one of them being significantly more influenced by the cryopreservation process. We obtained high correlations between head morphometry and SCSA parameters: standard deviation of DNA fragmentation index (SD-DFI) was correlated with perimeter and area ( r = 0.75 and r = 0.62, respectively) and DFIm and DFIt (moderate and total DNA fragmentation index) were correlated with perimeter ( r = 0.65 and r = 0.67, respectively). Nevertheless, classification of males according to SCSA or head morphometry did not completely agree so the two assays might explain male variability differently. We conclude that cryopreservation affected morphometry at least in a subset of spermatozoa. These results might improve future application of sperm banking techniques in this species.

Is abnormal sperm function an indicator among couples with recurrent pregnancy loss?

To determine whether or not sperm function parameters are altered in male partners of couples with a history of idiopathic recurrent pregnancy loss (RPL). In comparison with proven fertile volunteers, sperm function parameters like hypo-osmotic swelling (HOS), acrosomal status (AS), and nuclear chromatin decondensation (NCD) were assessed in vitro from male partners of couples with a history of idiopathic RPL. Infertility clinic and andrology laboratory at National Institute of Health and Family Welfare. Male partners of couples with a history of idiopathic RPL and proven fertile male volunteers (control). Standard semen analysis, assessment of sperm morphology, and sperm function with tests such as HOS, AS, and NCD. Sperm paameters, such as HOS, AS, and NCD, were assessed in semen samples from RPL in comparison with the proven fertile control group. Semen samples from the idiopathic RPL group showed below normal test scores in 57.1% of the cases for all three sperm parameters. The highest aberration (83% of cases) in sperm attributes was observed in NCD, followed by AS (45.7%) and HOS (42.9%). In contrast, abnormality in sperm morphology was limited to 5.7% of the cases. Subnormal sperm function is directly proportional with subnormal sperm motility (<50%) in 23% of the cases. Even in semen samples with normal sperm motility, sperm function scores were below normal in 31.4% of the RPL group. Reduction in test scores of sperm function, like HOS, AS, and NCD, in male partners of couples with idiopathic RPL suggests that sperms with altered or lowered functional competencies, if they fertilize the oocytes, may lead to the development of an unsustainable embryo resulting in early pregnancy loss. Normal sperm motility does not always ensure normal sperm function scores.

Sperm count and sperm motility in incidental high-grade varicocele

One hundred twenty-four otherwise healthy men with suspected high-grade varicocele, comprised of 28 patients with grade 2 and 96 with grade 3 varicocele, were referred to us by their primary care providers to determine their exact grade so that they could be exempted from the compulsory armed force service if they had grade 3 varicocele. In the grade 2 and grade 3 groups, mean sperm concentration and mean sperm motility were 80.89 million per milliliter (SD, 38.98 million/mL; median, 74.5 million/mL; range, 22-210 million/mL) and 78.77 million per milliliter (SD, 41.34 million/mL; median, 77 million/mL; range, 0-223 million/mL) and 58.39% (SD, 13.7%; median, 60%; range, 22-75%) and 60.31% (SD, 16.93%; median 65%; range, 0-80%), respectively, with no statistically significant differences between the two groups, no patient with infertility as a complaint during a 20-month period, and no patient with sperm concentrations of <20 million/mL, but with six patients with sperm motilities <50%. In summary, by far the great majority of patients with high grade varicocele have more than normal sperm output. Higher grades of varicocele are not associated with more pronounced deleterious effects on sperm concentration and percentage motility. Incidentally diagnosed high-grade varicocele is not a progressive condition, and almost all patients retain normal semen quality, at least over a period of 20 months.

Lack of Compliance by UK Andrology Laboratories With World Health Organization Recommendations for Sperm Morphology Assessment

Lack of Compliance by UK Andrology Laboratories With World Health Organization Recommendations for Sperm Morphology Assessment

P-546: Does sperm morphology by Kruger’s strict criteria have any relationship to blastocyst formation and clinical outcomes with ICSI?

Assesment of sperm morphology by Kruger’s strict criteria may be of little value in the clinical IVF setting. The increasing use of intracytoplasmic sperm injection (ICSI) for oocyte fertilization circumvents many problems associated with poor morphology such as poor zona binding, poor acrosome reaction, etc. Studies have suggested that poor sperm morphology is associated with decreased blastocyst formation. The objective of this study was to determine if critical assessment of sperm morphology during the pre-IVF semen analysis had any prognostic value as regards pregnancy outcome and embryo development.

Influence of staining and sampling procedures on goat sperm morphometry using the Sperm Class Analyzer

Computer-assisted sperm morphometry analysis (ASMA) has improved the assessment of sperm morphology, but the results depend on the use of adequate sampling and staining procedures of spermatozoa from individual species. In this study, the Sperm Class Analyzer® ASMA system was used for the morphometric analysis of goat sperm heads. Semen samples, obtained from four bucks, were used to evaluate the influence of three staining procedures (Diff-Quik, Hemacolor and Harris’ Haematoxylin) on the accuracy of image processing and sperm morphometry, the effect of the number of cells analysed and the repeatability of the method. These experiments were performed to obtain objective, accurate and reliable sperm morphometric measurements of goat spermatozoa.Diff-Quik and Harris’ Haematoxylin were significantly (p<0.05) more accurate than Hemacolor. However, Diff-Quik obtained the highest proportion of correctly analysed sperm heads (86.06%) and the lowest coefficients of variation on the image processing and morphometric measurements. The staining methods affected significantly the sperm dimensions (p<0.001) with increased values from Diff-Quik than Hemacolor and Harris’ Haematoxylin, respectively (Diff-Quik>Hemacolor>Harris’ Haematoxylin). No differences in morphometric parameters were found when 100, 150, 175 or 200 spermatozoa were analysed. The repeatability of results obtained was very high since no differences were found when measuring the same sperm on multiple attempts.In conclusion, to obtain objective, accurate and repeatable sperm morphometric measurements by the Sperm Class Analyzer® system in goats, the analysis of 100 spermatozoa from slides which have been previously stained with Diff-Quik is recommended.

Measuring mouse sperm parameters using a particle counter and sperm quality analyzer: A simple and inexpensive method

This study examined a method for analyzing the count, motility, and morphology of mouse epididymal sperm, optimizing the diluent, incubation time, sample concentration, and temperature, using a particle counter (CDA-500) to count and size sperm and a sperm quality analyzer (SQA-IIC) to measure sperm motility, quantified as the sperm motility index (SMI). The optimal conditions consisted of a 30-min incubation in D-MEM (Dulbecco's modified Eagle's medium; considering cost and availability) at 37 °C, with 5 × 10 6 cells mL −1 in the original solution. Furthermore, the influence of formalin fixation, and the correlation between the automated counter and a manual method were investigated. The sample fixation had no marked effect on the sperm count or morphology assessment. A linear correlation was observed between the manual and automated methods ( y = 0.920 x + 0.276; r 2 = 0.571; p < 0.001; range: (3–6) × 10 6). The suitability of the proposed method was confirmed using spermatozoa prepared from mice treated with the reproductive toxin diethylstilbestrol (DES). Using sperm from the cauda epididymidis on one side per mouse, we confirmed that measurement of these sperm parameters using the two devices was simple, rapid, inexpensive, and reproducible.

Morphological Image of Fresh and Cryopreserved Dog Semen Evaluated by the Strict Analysis of Sperm Morphology Method, Using Sperm Quality Analyzer (SQA IIc) Evaluation

Thirty fresh ejaculates from 15 dogs were cryopreserved in Tris-fructose-citric acid-egg-yolk extender with a glycerol content of 6%. Semen samples were examined by the methods of routine sperm analysis and by the SQA IIc device. The routine semen examination focused on the evaluation of parameters determining the quality of sperm membranes. The significance of monitoring semen quality in the course of the short-term survival test for predicting dog semen quality after thawing was assessed. Relevance of the assessment of sperm morphology, and above all the percentage of sperm with membrane changes in the acrosomal region was documented. The fact that the SQA device analyses semen quality by evaluating the mass of moving cells was confirmed. The results provided by the SQA IIc device appear insufficient for the needs of deeper dog semen analysis, especially morphology assessment.

The Relationship Between the Sperm Deformity Index (SDI), Apoptosis and Sperm Penetration Capacity

The Relationship Between the Sperm Deformity Index (SDI), Apoptosis and Sperm Penetration Capacity

Utilización del Integrated Semen Analysis System (ISAS) ® para el análisis morfométrico espermático humano y su significado en las técnicas de reproducción asistida

Aim The utility of a new computer-aided sperm morphology analyzer (the integrated semen analysis system [ISAS ®]) in predicting fertilization rates obtained with ejaculated spermatozoa from infertile men undergoing distinct assisted reproduction technologies (ART) was evaluated and compared with standard morphology analysis. Methods Ejaculates were obtained from men providing samples for intrauterine insemination (IUI) (n = 15), in vitro fertilization (IVF) (n = 20) and intracytoplasmic sperm injection (ICSI) (n = 19). Smears were prepared for routine morphological analysis using Papanicolaou staining and for assessment by ISAS ® using Hemacolor staining. Results Significant differences were found only in the sperm head length of ejaculated spermatozoa in patients undergoing IUI, IVF and ICSI. Spermatozoa selected by swim-up tended to have larger heads and shorter midpieces than spermatozoa found in the native ejaculate. No correlation was found between subjective sperm morphology and fertilization and pregnancy rates with the distinct ART used. In sharp contrast, a significant negative correlation was found between sperm head width, area and perimeter as assessed by ISAS ® after swim-up and the pregnancy rate for IUI and the fertilization rate for ICSI. No correlations were found with any of the parameters analyzed and the fertilization and pregnancy rates for IVF. Conclusion Sperm morphology assessment using ISAS ® may be useful when selecting patients for ART.

Lack of compliance by UK andrology laboratories with World Health Organization recommendations for sperm morphology assessment

Sperm morphology is known to correlate with the probability of conception both in vitro and in vivo, but the assessment of sperm morphology in the laboratory remains problematic. The 4th edition (1999) of the World Health Organization (WHO) Laboratory Manual has attempted to improve matters by giving rigorous recommendations regarding sperm morphology assessment. However, it is unknown how well these recommendations have been implemented in practice. A survey of the methods used to undertake the assessment of sperm morphology during semen analysis was undertaken in 37 laboratories in the UK. In total, only two laboratories (5%) were compliant with all current WHO guidelines regarding morphology assessment, including methods of staining and observation, classifying and sampling methods, and the participation in internal and external quality control programmes. These results illustrate an urgent need for education and training initiatives to encourage laboratories to become compliant with current WHO guidelines for sperm morphology assessment.

The establishment of sperm morphology satellite training laboratories in Africa

Summary Discrepancies in the results of semen analysis can be caused by a variety of factors, for example, a microscope for semen analysis can be situated in the practice of an urologist, cytologist or microbiologist, or even in a health care office in the countryside in a developing country. The study aimed to evaluate the (i) sperm morphology assessment skills of the participants who attended WHO seminology workshops and (ii) to appraise the possibility of using trained technologists to set up satellite training facilities in their own area. The Centre for Research in Reproductive Health, Ogun State University Teaching Hospital (Nigeria) initiated training sessions in their area. Subsequently, 16 technologists were trained in three sessions. Papanicolaou-stained slides, supplied by Tygerberg were used as test material. Pre- and post-training results were used to calculate the mean percentage of normal cells as well as the percentage deviation from the reference laboratory. The mean percentage normal spermatozoa recorded by the Nigerian trainees during pre-training evaluations differed by 38% from the reference value, compared with a post-training difference of 16% (P < 0.001). Tertiary academic hospitals can play an important educational role in the region. The present results underline the power of long distance educational programmes. Similar laboratories are currently being set up in Kenya and Zambia.

Effect of sample size and staining methods on stallion sperm morphometry by the Sperm Class Analyzer

Computer-assisted sperm morphometry analysis has improved the assessment of sperm morphology, but the results depend on the use of adequate evaluation and staining procedures of spermatozoa from individual species. In this study, the morphological module of the Sperm Class Analyzer&amp;reg;was used for the morphometric analysis of stallion sperm heads and midpieces. Semen samples were obtained from six fertile stallions in order to evaluate the influence of three staining procedures (Diff-Quik, Hemacolor and Harris&amp;rsquo; Haematoxylin) on the accuracy of image processing and sperm morphometry, and the effect of the sample size on sperm morphometric measurements. Harris&amp;rsquo; Haematoxylin was the staining technique of choice on the accuracy of the image processing with an optimum contrast of sperm cells with the surrounding background that allows an efficient boundary detection and segmentation which results in the highest proportion of sperm heads and midpieces assessed (80.47%). The results indicate that the staining methods affected significantly the sperm dimensions with increased values from Diff-Quik than Hemacolor and Harris&amp;rsquo; Haematoxylin respectively (Diff-Quik &amp;gt; Hemacolor &amp;gt; Harris&amp;rsquo; Haematoxylin). No differences in morphometric parameters were found when 100, 150, 175 or 200 spermatozoa were analysed. In conclusion, to obtain objective and accurate sperm morphometric measurements by the Sperm Class Analyzer&amp;reg; system in the stallion, it&amp;rsquo;s recommended the analysis of 100 spermatozoa from slides which have been previously stained with Harris&amp;rsquo; Haematoxylin.

Individual assessment of sperm morphology of single spermatozoa used for intracytoplasmic sperm injection

Intracytoplasmic sperm injection (ICSI) is an integral part of assisted reproduction. Although many papers have shown that global sperm count, sperm motility and sperm morphology of the ejaculate play no role in the fertilization rate after ICSI, embryologists who carry out ICSI, try to use the 'best looking' spermatozoa. The aim of the study was to investigate whether those spermatozoa with the best morphology really achieve the highest fertilization rate. In the present study, a total of 798 spermatozoa used for ICSI were documented by high-resolution photo. After ICSI the oocytes were cultured in single droplets and the formation of pronuclei was assessed 16 h later. The spermatozoa (all normal according to WHO criteria) were classified into four groups of different morphology. Group 1: normal head shape (approximately 5 microm diameter), group 2: like group 1, but with 15-20% smaller diameter, group 3: like group 1, but with 15-20% larger diameter, and group 4: like group 1, but with slight mid-piece cytoplasmic irregularities. Using the Pearson chi-square test, no significant difference in terms of fertilization was found among the different groups, showing that marginal sperm differences do not alter the fertilization process in ICSI.

Novel associations between specific sperm morphological defects and leukocytospermia

To examine the relationship between leukocyte concentrations in semen and sperm morphology in a group of infertile men and healthy fertile donors. A prospective clinical study. Male infertility clinic at a tertiary care teaching hospital and a reproductive medicine unit at a Women's Hospital in the United Kingdom. Fifty-six infertile men and 13 healthy fertile sperm donors (control). Standard semen analysis, seminal leukocyte concentration, and the assessment of sperm morphology and sperm deformity index (SDI), applying Tygerberg's strict criteria. Granulocyte concentrations in semen, percentages of different sperm morphological abnormalities, and SDI scores. Leukocyte concentrations were statistically significantly and negatively correlated with the proportion of sperm with damaged acrosomes, cytoplasmic droplet, tail defects, and SDI scores with normal and borderline morphology. The percentage sperm motility was significantly and negatively correlated with leukocytic concentration in semen. However, the leukocytic concentration was not significantly correlated with sperm concentration. This is the first study to report a significant positive correlation between leukocytospermia and sperm tail defects, acrosomal damage, and high SDI scores. These observations suggest that leukocytospermia is associated with compromised sperm structural integrity.

A novel association between sperm deformity index and oxidative stress-induced DNA damage in infertile male patients

Sperm deformity index (SDI) is a novel quantitative expression of sperm morphological quality that enhances both the predictive power and reproducibility of sperm morphology assessment. Morphologically abnormal spermatozoa are rich in cytoplasmic droplets, which contain NADPH oxidase capable of mediating oxidative stress (OS). The objective of our study was to investigate the impact of abnormal sperm morphology using SDI on NADPH-mediated reactive oxygen species (ROS) production and its correlation with sperm DNA damage. Prospective controlled study. Semen samples were collected from 6 healthy donors with normal standard semen parameters according to WHO guidelines and 7 infertile male patients. Mature sperm were isolated using double density gradient (ISolate, 47% and 90%), re-suspended in BWW media and subdivided into 2 aliquots; the first was incubated with 5 mM NADPH up to 24 hours, while the second subset was incubated without NADPH to serve as control. Sperm morphology was assessed by strict Tygerberg's criteria. The SDI was calculated by dividing the total number of deformities observed by the number of sperm evaluated. Sperm DNA damage was assessed using terminal deoxynucleotidyl transferase-mediated fluorescein-dUTP nick end labeling (TUNEL) assay by flow cytometry and ROS levels (x 106 counted photons per minute) by chemiluminescence assay using lucigenin as a probe. The median and interquartile values (25thand 75th percentiles) of SDI were higher in patients compared to donors both in whole ejaculate[1.89 (1.63, 2.17) vs. 1.62 (1.53, 1.8), P = 0.03)] as well as in mature spermatozoa[2 (1.8, 2.1) vs. 1.53 (1.52, 1.58), P = 0.008]. Following NADPH incubation, a significantly higher DNA damage was seen after 24 hours in patients compared with donors[16.56 (11.29, 40) vs. 4.4 (3.92, 5.25), P = 0.007]. Similarly, in controls, DNA damage was higher in patients compared with donors[5.1 (3.87, 7.74) vs. 1.79 (2.87, 3.36), P = 0.03]. However, combining aliquots treated with NADPH (patients and donors) also showed higher incidence of DNA damage than those not treated[10 (4.69, 24.85) vs. 3.85 (2.58, 5.1), P = 0.008]. The increase in ROS levels was also more pronounced in aliquots exposed to NADPH[1.22 (0.3, 1.87) vs. 0.39 (0.1, 0.57), P = 0.03). SDI correlated with the percentage increase in sperm DNA damage following incubation for 24 hours (r = 0.5, P = 0.009). SDI is a useful tool in identifying infertile male patients with abnormal prevalence of OS-induced DNA damage. NAPDH plays a role in ROS-mediated sperm DNA damage, which appears to be more evident in infertile patients with semen samples containing high incidence of morphologically abnormal spermatozoa.

Novel association between sperm reactive oxygen species production, sperm morphological defects, and the sperm deformity index

Objective To examine the relationship between sperm reactive oxygen species (ROS) production and sperm morphology in a group of infertile men and healthy fertile donors. Design A prospective clinical study. Setting Male infertility clinic, Glickman Urological Institute, The Cleveland Clinic Foundation, Cleveland, Ohio, and the Reproductive Medicine Unit, Liverpool Women's Hospital, United Kingdom Patient(s) Thirty-nine infertile men and 13 healthy fertile donors (control). Intervention(s) Standard semen analysis, seminal leukocyte concentration, assessment of sperm morphology, and measurement of sperm ROS production. Main outcome measure(s) Levels of sperm ROS production, percentages of different sperm morphological abnormalities, and the sperm deformity index (SDI) scores. Result(s) A significant negative correlation was observed between sperm ROS production and the proportion of sperm with normal morphology and borderline morphology. Reactive oxygen species production was positively correlated with the proportion of sperm with amorphous heads, damaged acrosomes, midpiece defects, cytoplasmic droplets, tail defects, and SDI scores. Logistic regression analysis identified a two-variable model including SDI and percentage sperm motility, which correctly identified 84% of individuals with high seminal ROS and 85% of individuals with low seminal ROS. The model had an overall accuracy of 85%. Conclusion(s) The standard semen analysis to assess sperm motility, sperm morphology, and the SDI scores is a useful tool in identifying infertile men with high seminal ROS in infertility clinics where facilities for measuring levels of seminal ROS are not available.