Evaluation Of Sperm Concentration Research Articles

P–053 Concentration of bisphenol S in human seminal plasma negatively correlates with decreasing sperm concentration in ejaculate

Abstract Study question Is there a relationship between concentration of bisphenol S in seminal fluid and spermiogram parameters? Summary answer Bisphenol S was detected in 81% of seminal plasma samples. Negative correlation was found between BPS concentration and total sperm count in normozoospermic men. What is known already Human spermatogenesis can be influenced by a range of chemicals present in our environment. Bisphenol S (BPS) is a very frequent compound commonly used as a softener in production of plastics, where it has replaced bisphenol A. It is an endocrine disruptor frequently associated with negative effects on reproduction. It has been observed that BPS can affect testicular development in rodent males. In addition, it has cytotoxic, reprotoxic and neurotoxic effects and induces the oxidative stress bringing negative effects on spermatogenesis. BPS has been detected in food, drinks or cosmetics. Its direct effect on spermatozoa or spermatogenesis is still unclear. Study design, size, duration A total of 38 patients (25 normozoospermic) aged 24 to 42 years, non-smokers, with BMI between 19,9–32,9 were included in this prospective study from 2018 to 2020. None of them had varicocele, urogenital infections or other urological problems. Their seminal plasma was separated by centrifugation. BPS was extracted using solvent extraction followed by preconcentration step. The samples were analysed on Agilent 6495 Triple Quadrupole. Two MS/MS transitions were used for quantitative LC-MS/MS analyses. Participants/materials, setting, methods All the men included in this study signed an informed consent and agreed with analyses of their samples. These analyses were approved by Ethical committee of University Hospital Brno. We evaluated the relationship between concentration of BPS in seminal plasma, sperm concentration, total sperm count, total motility, progressive motility, morphology and fragmentation of DNA in spermatozoa. Statistical evaluation was performed by one individual one-dimensional regression model (p-value lower than 0.05 were considered as statistically significant). Main results and the role of chance The examination revealed the presence of BPS in 31 samples of seminal plasma (81% of all the samples). In 6 samples, the concentration was under level of detection and in one sample under level of quantification. In 7 samples, a very high concentration was detected (>0.1 ng/ml). These values were then compared to spermiogram parameters and sperm DNA integrity. There were no significant differences between the concentration of BPS and morphology of spermatozoa, progressive motility and total motility. In case of the DNA integrity, the opposite trend was observed, lower proportions of spermatozoa with fragmented DNA were found in samples with higher concentrations of BPS. Evaluation of sperm concentration and BPS concentration showed relationship of increasing BPS concentration with significantly lower sperm concentration, the differences were most obvious when only the normozoospermic men were compared. Evaluation of BPS concentration and total sperm count revealed the same trend with statistically significant difference in the category of normozoospermic men. Due to the small number of samples, a negative effect of extreme values on the statistical evaluation cannot be excluded. Currently, more analyses focused on detection of BPS in seminal plasma are carried out in order to obtain sufficiently larger data set. Limitations, reasons for caution A limitation is the number of samples included and analysed in this study, which slightly reduced the power of statistical analysis. Wider implications of the findings: These results document that BPS was present in 81% of analysed samples. Knowing the concentration of BPS in seminal fluid is important for understandig of impact of BPS on male fertility. Our future work will be focused on detection of other bisphenols in seminal plasma. Trial registration number MH CZ – DRO (FNBr, 65269705), AZV NV18–01–00544, Czech Ministry of Education, Youth and Sports (CZ.02.2.69/0.0/0.0/19_074/0012727)

Microfabrication of low-cost customisable counting chambers for standardised estimation of sperm concentration.

Evaluation of sperm concentration is essential for research and procedures involving AI, cryopreservation and sperm quality assessment. Microfabrication technologies have shown tremendous potential for rapid prototyping and fabrication of devices to assist reproduction and fertility research, but such utility has not yet been made available for most reproduction laboratories. The aim of this study was to evaluate the feasibility of using microfabrication techniques to produce counting chambers for estimation of sperm concentration. Zebrafish (Danio rerio) spermatozoa were used as a model for evaluation of functionality of the chambers. These microfabricated enumeration grid chambers (MEGC) were composed of a polydimethylsiloxane (PDMS) coverslip with grid patterns (100 μm×100 μm) and a PDMS base platform to create a known volume with a 10-μm height to restrict the cells to a single layer. The results of cell counts estimated by two of three prototype MEGC devices tested were not significantly different from the control device, a commercially available Makler chamber. The material cost for a MEGC was less than US$0.10 compared with product costs of approximately US$100 for a standard haemocytometer and US$700 for a Makler counting chamber. This study demonstrates the feasibility of microfabrication in creating low-cost counting chambers to enhance standardisation and strengthen interdisciplinary collaborations.

The Value of Sperm Velocity, Strict Morphology and the Hypo-osmotic Swelling Test as Predictors of the Sperm Fertilization Potential

Although prevalence of infertility is high and are predominantly or partly due to a male factor, the diagnostic tools in male fertility are insufficient being mainly based on the evaluation of sperm concentration, motility and morphology . These parameters are, however, poorly standardized, subjective) and not powerful predictors of fertility. The aim of this work was to evaluate the sperm velocity characteristics, the sperm morphology assessment determined by strict criteria and the hypo-osmotic swelling (HOS) test as predictors of sperm fertilization potential. A comparison of 3 parameters to determine which of them is a better predictor of the fertilization potential of a given semen sample in the IVF model, namely the sperm velocity characteristics determined by computerised semen analysis, strict sperm morphology and the HOS test. Data from 58 couples attending the assisted conception unit for conventional in-vitro fertilization (IVF) treatment were analyzed. Simple linear regression and multiple stepwise regression analysis were performed taking the fertilization rate as the dependent variable and the following as independent variables: (1) female partner's age, (2) number of human menopausal gonadotropin (hMG) ampoules, (3) sperm count, (4) grade A motility, (5) grade B motility, (6) grade C motility, (7) grade A+B motility, (8) mean sperm velocity, (9) linear velocity, (10) linearity index, (11) strict morphology and (12) the HOS test. The results were used to calculate the cut-off levels for the statistically significant predictors of fertilization. Only the mean sperm velocity and the strict sperm morphology assessment showed significant correlation with the fertilization rate. The cut-off values were a mean sperm velocity of 16 μm/sec and a strict sperm morphology of 10 %. A minimum sperm velocity of 13 μm/sec and a minimum strict sperm morphology of 2 % were necessary to achieve fertilization in 50% of the oocytes. The results of the HOS test did not correlate with the fertilization rate. Sperm velocity and strict sperm morphology evaluation are good predictors of fertilization potential. The HOS test is not a good predictor of fertilization potential.

Semenology training programs: 8 years' experience

To evaluate the immediate effect of hands-on training on the quality of technical skills of laboratory technicians. Retrospective analytical study. Academic institutions and private infertility clinics. One hundred and ten laboratory technicians. None. First, 110 participants from 16 African countries attended 5-day semenology workshops at Tygerberg Hospital. During these workshops the methodology as described by the World Health Organization manual for the analysis of human semen, namely, sperm concentration, motility, vitality, and morphology, formed part of the curriculum. Second, two experienced morphology readers from the Tygerberg group presented 23 1-day sperm morphology workshops in nine other countries. The semenology workshops indicated a significant improvement in the evaluation of sperm concentration only, whereas pretraining and posttraining results for motility did not differ. Vitality reports did not differ from those of the experienced worker in the first place. Calculation by means of a linear regression model showed a significant decrease in the mean posttraining scores from the pretraining scores for the sperm morphology courses. Training improved technical skills for the evaluation of morphology and sperm concentration. The evaluation of graded sperm motility seems to be more difficult to teach over a short period.

Evaluation of Sperm Concentration by the Hemacytometer Method: Comparison of Four Counting Fluids

Evaluation of Sperm Concentration by the Hemacytometer Method: Comparison of Four Counting Fluids