The impact of cryopreservation on the morphology of spermatozoa in men with oligoasthenoteratozoospermia.

The cryopreservation of ejaculate can reduce the viability, motility, and morphological characteristics of the spermatozoa of infertile men. Oligoasthenoteratozoospermia (OAT) is the most common cause of male subfertility. The aim of this study was to evaluate the morphological characteristics and viability of progressive motile sperm fraction before and after cryopreservation, and to determine whether cryopreservation of progressive motile sperm fraction is effective in eliminating morphologically abnormal sperm in men with OAT. An increased proportion of spermatozoa with normal morphology in fresh progressive motile sperm fraction compared with fresh ejaculate has been observed. After cryopreservation, the motility was 65.5 ± 8.8%; the proportion of spermatozoa with normal morphology increased non-significantly compared with freshly prepared motile sperm fraction (35.6 ± 5.5%). Concurrently, the proportion of cryopreserved spermatozoa with head defects increased significantly by 1.7 times (to 38.4 ± 4.7%) and the proportion of almost all morphologically abnormal sperm cells, particularly spermatozoa with multiple abnormalities, was reduced significantly. These data appear to be a novel finding in the context of patients with OAT. Using such spermatozoa for in vitro fertilization leads to a significant decrease in both a number of embryos at the cleavage stage and the blastocysts formation rate. High-magnification sperm morphology examination and selection, IMSI, post-cryopreservation significantly increased the likelihood of successful oocyte fertilization and subsequent embryo development.

Magnetic-activated cell sorting of non-apoptotic spermatozoa improves the quality of embryos according to female age: a prospective sibling oocyte study.

PURPOSE: The main aim of our study was to evaluate the benefit of the use of non-apoptotic spermatozoa selected by magnetic-activated cell sorting (MACS) for ICSI procedures for couples in which the women had good prognoses and the male factor of infertility was teratozoospermia. METHODS: Twenty-six couples were treated with ICSI after MACS selection of non-apoptotic spermatozoa following a sibling oocyte approach. Half of the oocytes were microinjected with conventionally prepared spermatozoa, and the other half were microinjected with non-apoptotic, MACS-selected spermatozoa. To assess the influence of MACS selection of spermatozoa on the outcomes of the ICSI cycles, the fertilization, embryo quality, pregnancy, and delivery rates were evaluated and compared between the sibling oocyte groups. RESULTS: When subpopulations of couples according to female age were analyzed, a significant difference in quality of blastocyst was observed. More precisely, in a group that was treated with MACS-ICSI, a higher percentage of good quality blastocysts was found among women older than 30 years (75.0 vs. 33.3%; P = 0.028), while there was no difference among younger women. If all included couples were compared regardless of age, no significant difference was observed in the outcome of the ICSI/MACS-ICSI cycles in terms of oocytes and embryos. Additionally, after the ICSI and MACS-ICSI procedures, the morphologies of the prepared spermatozoa were compared. Results showed that the overall percentage of morphologically normal spermatozoa did not differ significantly between the ICSI and MACS-ICSI procedures. However, detailed analyses of the morphologically abnormal spermatozoa revealed significantly more spermatozoa with abnormal tails after MACS-ICSI procedure, which may be potential consequence of the selection procedure. Moreover, the trends towards less spermatozoa with abnormal heads and towards more spermatozoa with abnormal necks and midpieces after MACS-ICSI procedure were revealed, although the differences were not significant. CONCLUSIONS: Couples dealing with male infertility due to teratozoospermia can benefit from MACS selection of spermatozoa with higher percentage of good quality blastocysts but only when the woman is older than 30 years.

Different Nuclear Architecture in Human Sperm According to Their Morphology.

Human sperm parameters serve as a first step in diagnosing male infertility, but not in determining the potential for successful pregnancy during assisted reproductive technologies (ARTs) procedures. Here, we investigated the relationship between sperm head morphology at high magnification, based on strict morphologic criteria, and the nuclear architecture analyzed by fluorescence in situ hybridization (FISH). We included five men. Two of them had an elevated high-magnification morphology score of 6 points (Score 6) indicating high fertility potential, whereas three had a low score of 0 points (Score 0), indicating low fertility potential. We used FISH to study the inter-telomeric distance and the chromosomal territory area of chromosome 1 (Chr. 1). We then compared these two parameters between subjects with high and low scores. FISH data analysis showed that the inter-telomeric distance (ITD) and chromosomal territory area (CTA) of Chr. 1 were significantly higher in subjects with low scores (score 0) than high scores (score 6). Our results suggest that (i) there is a link between nuclear architecture and sperm head abnormalities, particularly vacuoles; and (ii) it is possible to select spermatozoa with normal nuclear architecture, which might indirectly explain the positive ART outcomes observed with this technique.

The development of new biomarkers of spermatozoa quality in cattle.

There is a current need for new biomarkers of spermatozoa quality, that consistently and correctly identify spermatozoa that will successfully contribute to subsequent embryo development. This could improve the standardization of semen analysis, decrease early embryo mortality, and use these biomarkers as a selection tool before servicing females. This study utilized imaging techniques to identify potential biomarkers of sperm quality, using sires previously classified as high (n = 4) or low (n = 4) performing at producing blastocysts in vitro. Spermatozoa were assessed before and following a gradient purification protocol, to understand how populations of cells are impacted by such protocols and may differ between in vivo and in vitro use. Pre-gradient samples from low-performing sires had an increased incidence of DNA damage, although post-gradient samples from high-performing sires were found to have an increased incidence of DNA damage. When evaluating morphology via fluorescent microscopy, the most prevalent defects in pre-gradient samples from high-performing sires were tail defects, which are successfully removed during purification processing. The most prevalent defects in pre-gradient samples from low-performing sires were aggresome defects located in the head, which would be brought into an oocyte upon fertilization and may impair embryo development. Image-based flow cytometry (IBFC) was employed to quantify defect prevalence to evaluate a greater sample size decreasing the variability that exists in manual assessments. Using IBFC, aggresome defects were again identified in the heads of spermatozoa from low-performing sires. Post-gradient samples from low-performing sires had a significantly greater (p < 0.05) incidence of aggresome defects than post-gradient samples from high-performing sires. Additionally, IBFC was used to evaluate spermatozoa viability following gradient purification. Distinct populations of sperm cells were identified. High-performing sires had more spermatozoa in the population deemed most viable than low-performing sires. This study demonstrated that spermatozoa defects vary in populations before and following gradient purification, indicating that it may be beneficial to separately evaluate semen for in vivo and in vitro use. Furthermore, a prevalent defect in low-performing sires that could explain a discrepancy between successful fertilization and embryo development was identified. Therefore, elucidating a malfunction regulated by sire, that could potentially affect early embryo development.

Molecular Alterations and Severe Abnormalities in Spermatozoa of Young Men Living in the "Valley of Sacco River" (Latium, Italy): A Preliminary Study.

The Valley of Sacco River (VSR) (Latium, Italy) is an area with large-scale industrial chemical production that has led over time to significant contamination of soil and groundwater with various industrial pollutants, such as organic pesticides, dioxins, organic solvents, heavy metals, and particularly, volatile organic compounds (VOCs). In the present study, we investigated the potential impact of VOCs on the spermatozoa of healthy young males living in the VSR, given the prevalent presence of several VOCs in the semen of these individuals. To accomplish this, spermiograms were conducted followed by molecular analyses to assess the content of sperm nuclear basic proteins (SNBPs) in addition to the protamine-histone ratio and DNA binding of these proteins. We found drastic alterations in the spermatozoa of these young males living in the VSR. Alterations were seen in sperm morphology, sperm motility, sperm count, and protamine/histone ratios, and included significant reductions in SNBP-DNA binding capacity. Our results provide preliminary indications of a possible correlation between the observed alterations and the presence of specific VOCs.

Change in Morphology of Spermatozoa from Dismount Semen during the Breeding Season in Thoroughbred Stallions in Japan.

To clarify the physiological changes of sperm morphology in active Thoroughbred stallions during the breeding season, we examined the dismount semen collected from the penile urethra immediately after service. The spermatozoa were analyzed for relationships between the morphology and the stallion's age or the number of services. Seasonal variation was apparent in the rate of the sperm tail abnormalities, spermatozoa with cytoplasmic droplets, appearance of medusa cells, and sperm head length. Area and width of the sperm head correlated negatively with age (P<0.05). The rate of appearance of medusa cells and the length of the sperm head were positively related to the number of services (P<0.05), and the aspect ratio was negatively related (P<0.01).

Gephyrocharax machadoi, a new species of Stevardiinae (Characiformes: Characidae) from the Rio Paraguai basin, central Brazil.

Gephyrocharax machadoi, new species, is described from tributaries of the rio Sepotuba, upper rio Paraguai basin, Mato Grosso state, Brazil. The new species is distinguished from its congeners, except Gephyrocharax major, by presenting two modified scales on the ventral caudal-fin lobe (pouch scales): a larger, sexually dimorphic scale, slightly superior and anterior in position, followed by a smaller, posterior and ventrally placed accessory scale (vs. a single modified pouch scale without ventrally placed accessory scales). Gephyrocharax machadoi differs from G. major by presenting a gap (more conspicuous in mature males than in mature females) between the second and third ventral procurrent caudal-fin rays (vs. second and third ventral procurrent caudal-fin rays near each other or fused distally); premaxilla with tricuspid teeth (vs. tetra- to pentacuspid teeth); and body depth 21.5-25.4% of SL (vs. 25.9-36.8% in SL). Spermiogenesis and spermatozoa morphology are also described.