Obesity represents a growing problem due to its impacts on human health and reproduction. In this study, we analysed semen quality, sperm DNA integrity and gene-specific CpG methylation in 116 healthy men from normal population. The men were divided into three groups according to their body mass index (BMI), and their ejaculates were analysed using standard methods, sperm chromatin structure assay (SCSA), methylation next generation sequencing (NGS) and amplicon sequencing. The sperm methylation NGS revealed six significantly differentially methylated regions (DMRs). Using subsequent targeted amplicon sequencing in 116 men, two of the DMRs were proved as differentially methylated in sperm of men with normal BMI vs. BMI ≥ 25. The DMRs were located in the EPHA8 and ANKRD11 gene. Also, we detected a significant decline in the EPHA8, ANKRD11 and CFAP46 gene methylation in association with increasing BMI values. The genes EPHA8 and ANKRD11 are involved in the nervous system and brain development; the CFAP46 gene plays a role in a flagellar assembly and is associated with sperm motility. Significantly lower rates of motile and progressive motile sperm were observed in men with BMI ≥ 30. Our results show that excess body weight can modify CpG methylation of specific genes, affect sperm motility, and compromise sperm chromatin integrity. These factors can stand behind the observed reduced fertility in men with obesity. The methylation changes might be transmitted to their offspring through sperm, and become a basis for possible developmental and reproductive issues in the next generation.

• Klíčová slova
• BMI, Body mass index, DNA fragmentation, DNA methylation, Obesity, Semen, Sperm,
• MeSH
• analýza spermatu * MeSH
• chromatin * metabolismus   MeSH
• CpG ostrůvky MeSH
• dospělí MeSH
• index tělesné hmotnosti * MeSH
• lidé MeSH
• metylace DNA * MeSH
• mladý dospělý MeSH
• motilita spermií genetika   MeSH
• obezita genetika   MeSH
• spermie * metabolismus   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• chromatin * MeSH

Spermatozoan quality can be evaluated in different ways, here we focus on the analysis of DNA, RNA and epigenetic status of germ cells. These characterizations also can be the bases for explaining sperm quality at other levels, so we will see how some of these molecules could affect other sperm quality markers. Moreover, we consider the possibility of using some of these molecules as predictors of sperm quality in terms of the ability to produce healthy offspring. The relevant effect of different types of RNA molecules in germ line specification and spermatogenesis and the importance of germ cell DNA integrity and a proper epigenetic pattern will be also discussed. Although most studies at this level have been performed in mammals, some information is available for fish; these recent discoveries in fish models are included. We provide a general overview on how these molecules could have a deep influence in the final sperm quality.

• Klíčová slova
• DNA, Epigenetics, Germ cells, Non-coding RNAs, Sperm quality, mRNAs,
• MeSH
• DNA genetika   MeSH
• epigeneze genetická fyziologie   MeSH
• RNA genetika   MeSH
• ryby genetika   fyziologie   MeSH
• spermatogeneze genetika   fyziologie   MeSH
• spermie fyziologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• DNA MeSH
• RNA MeSH

BACKGROUND: We examined sperm quality in a cohort of city policemen in Ostrava at the end of a period with high concentrations of air pollutants (winter) and in the same cohort at the end of a relatively low exposure period (summer). METHODS: The study group was comprised of 54 nonsmoking city policemen living and working in Ostrava, Czech Republic. Average daily air-pollutant concentrations recorded by stationary monitoring for 90 days preceding the collection of semen samples were evaluated for different city districts of Ostrava. Standard semen parameters were assessed according to the guidelines of the World Health Organization (2010). The parameters were semen volume, sperm concentration, sperm morphology, sperm motility, acrosome reaction and sperm plasma membrane integrity. Sperm DNA damage was analysed by the sperm chromatin structure assay (SCSA). Sperm motion characteristics were determined by Computer Assisted Semen Analysis (CASA). RESULTS: The concentrations of all monitored pollutants (particulate matter, sulphur dioxide, nitrogen oxides, carbon monoxide, benzo[a]pyrene, benzene) were significantly increased during winter (p < 0.001), except for ozone, the concentration of which was significantly higher during summer. Sperm volume, concentration, % vitality, % sperm morphology (normal form) and % acrosome-intact sperm did not differ significantly between the monitoring periods. The percentages of total motility and progressive motility were significantly higher in March, i.e. at the end of winter (p = 0.001). However, CASA testing showed differences in sperm motion kinetics between spring and autumn samples. In the spring samples, we found a significantly lower % of straightness (p = 0.044) and the length of straight-line path (p = 0.01), while linearity and straight-line velocity were near the borderline value (p = 0.064; p = 0.054, respectively). As compared to summer, high exposure to air pollution during winter significantly increased the extent of sperm chromatin integrity damage (median 22.6 vs. 18.6%) (p = 0.003) and the proportion of immature spermatozoa (median 11.2 vs. 9.9%, p = 0.001). Sperm DNA damage negatively correlated with total motility and progressive motility (r = -0.611, -0.299; p < 0.001). The negative correlation with vitality, normal morphology and acrosome-intact sperm (r = -0.522, -0.550 and -0.511, respectively) was also significant (p < 0.001). CONCLUSION: The examination of the same cohort of city policemen at the end of a period of high air pollution and at the end of relatively low exposure reduced the effects of age, different lifestyles, different occupational exposures, localities and genetic polymorphism on sperm quality impairment associated with air pollution. This study did not demonstrate impaired standard semen parameters in association with exposure. It was shown that sperm chromatin damage and the percentage of immature sperm were highly sensitive to air pollution.

• Klíčová slova
• Air pollution, DNA fragmentation, Occupational exposure, Sperm quality,
• MeSH
• analýza spermatu * MeSH
• DNA MeSH
• lidé MeSH
• motilita spermií MeSH
• sperma MeSH
• spermie MeSH
• znečištění ovzduší * statistika a číselné údaje   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA MeSH

Chromatin remodeling, including histone post-translational modifications, during spermatogenesis can affect sperm quality and fertility, and epigenetic marks may therefore be useful for clinical evaluations of sperm. Together with histone hyperacetylation, the dimethylation of histone H3 on lysine K4 (H3K4me2) is also required during protamination. Accordingly, we evaluated the utilization of this epigenetic mark for the identification of sperm with decrease quality and immature chromatin. In this study, 99 semen samples, including 22 normozoospermic (N), 63 asthenozoospermic (A), and 14 oligoasthenozoospermic (OA) samples, were comprehensively analyzed with respect to H3K4me2 levels, DNA damage (DNA fragmentation index, DFI), and sperm immaturity (high DNA stainability, %HDS), as determined by a sperm chromatin structure assay using flow cytometry. We detected a significant relationship between H3K4me2 and %HDS (r = 0.47; p < 0.001). Furthermore, we observed negative correlations between H3K4me2 and sperm concentration, motility, and mitochondrial activity (p < 0.05). The increase in immaturity as semen quality decreased (N > A > OA) indicates the importance of chromatin immaturity and histone code deviations in sperm evaluations. Using various approaches, our study elucidated H3K4me2 as a molecular marker of sperm quality with potential use in reproductive medicine.Abbreviations: A: asthenozoospermic; AO: acridine orange; ART: assisted reproductive therapy; BWW: Biggers-Whitten Whittingham; DAPI: 4',6' -diamidino-2-phenylindole; DFI: DNA fragmentation index; H3K4me2: dimethylation of lysine K4 on histones H3; HDS: high DNA stainability; HRP: horseradish peroxidase; MACS: magnetic-activated cell sorting; N: normospermic; NGS: normal goat serum; OA: oligoasthenozoospermic; PTM: post-translational modification; SCSA: sperm chromatin structure assay; SUTI: sperm ubiquitin tag assay; TBS-T: TBS with 0.5% Tween-20.

• Klíčová slova
• H3K4me2, Sperm DNA, chromatin immaturity, epigenetics, high DNA stainability,
• MeSH
• analýza spermatu MeSH
• astenozoospermie metabolismus   MeSH
• biologické markery metabolismus   MeSH
• dospělí MeSH
• histonový kód * MeSH
• histony metabolismus   MeSH
• lidé MeSH
• metylace MeSH
• oligospermie metabolismus   MeSH
• restrukturace chromatinu * MeSH
• spermie metabolismus   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biologické markery MeSH
• histony MeSH

Improvement of sperm quality with low motility by storage could ensure higher success of fertilization and maintain higher genetic diversity, especially for sturgeons, which as endangered species have limited broodstock and gametes. Sperm was collected from mature male sterlet Acipenser ruthenus and motility was evaluated using the CASA system; samples were categorized as GS 'good sperm' (>80%) or BS 'bad sperm' (<20%). Samples from both groups were incubated with seminal plasma from good- (GSP) and bad-quality sperm (BSP), respectively for 15 min, 6 h, 24 h and 96 h at 4 °C. Motility of BS incubated in GSP increased after different storage times compared to BS incubated in BSP, while the motility and velocity of GS incubated in BSP decreased compared to GS incubated in GSP. Fertilization rates were evaluated with samples stored for 15 min and 6 h post-stripping; fertilization and hatching rate of BS after incubation in GSP increased significantly compared to the BS incubated in BSP. Inorganic ion (Na+, K+, Cl-) concentrations and osmolality of BSP were significantly lower than that of GSP. These results indicated that sterlet sperm quality can be revitalized by incubation with GSP. Further, fertilization capacity of BS after incubation in GSP can reach similar levels to the good quality sperm (∼70%). Low ion concentration and osmolality in BSP may be a partial cause of low sperm quality. The current study is the first report on the capability to revitalize low quality sterlet sperm by storage in GSP.

• Klíčová slova
• Fertilization success, Motility, Seminal plasma, Sperm storage, Sturgeon,
• MeSH
• fertilizace MeSH
• motilita spermií * MeSH
• ryby MeSH
• sperma MeSH
• spermie MeSH
• uchování spermatu * veterinární   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Human populations living in urban industrial regions of developed countries are exposed to high levels of environmental pollutants. The reproductive consequences of the exposure to air pollution can be monitored through semen analysis and molecular methods. In this study, we tested the possible impact of seasonal changes in the level of air pollution on the semen quality and sperm DNA methylation of 24 men living and working in the industrial agglomeration of Ostrava (Czech Republic). The study participants were healthy non-smokers. The study group was homogeneous regarding their profession, moderate alcohol consumption, no drug abuse and no additional exposure to chemical toxicants. We performed targeted methylation next generation sequencing (NGS) using Agilent SureSelect Human Methyl-Seq and Illumina NextSeq 500 platform to analyze semen samples collected repeatedly from the same men following the season of high (winter) and low (summer) air pollution exposure. We did not detect any adverse effects of the increased exposure on the semen quality; neither we found any difference in average sperm DNA methylation between the two sampling periods. Our search for differentially methylated CpG sites did not reveal any specific CpG methylation change. Our data indicate that the seasonal changes in the level of the air pollution probably do not have any substantial effect on sperm DNA methylation of men living in the highly polluted industrial agglomeration for a long period of time.

• Klíčová slova
• DNA methylation, air pollution, epigenetics, exposure, sperm,
• MeSH
• analýza spermatu * MeSH
• lidé MeSH
• metylace DNA genetika   MeSH
• průmysl MeSH
• spermie MeSH
• znečištění ovzduší * škodlivé účinky   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The use of microfluidic sperm sorting (MFSS) systems in infertility treatment is increasing due to their practicality and ease of use. While often presented as highly effective, their efficacy in patients with varying sperm analysis results remains uncertain. In this study, we evaluated the effectiveness of MFSS compared with the swim-up (SU) technique in terms of oxygen radical levels and spermiogram parameters. Samples from each patient were processed using both methods, followed by assessments of sperm concentration, motility, morphology, DNA integrity, acrosomal status, and mitochondrial membrane potential. Participants were selected based on sperm analysis and categorized as normozoospermic (n = 40) or non-normozoospermic (n = 28). An analysis of separation techniques revealed no significant differences, except for a lower percentage of DNA-fragmented sperm in the MFSS group compared with SU within the non-normozoospermic cohort (SU: 10.0% vs. MFSS: 5.69%, p = 0.027). No differences were observed between SU and MFSS in normozoospermic men. The MFSS method is a simple technique, frequently used in laboratories, that yields good results but does not offer a substantial advantage over SU. The primary benefit of MFSS appears to be a significant reduction in the proportion of sperm with DNA fragmentation compared with SU in patients with abnormal sperm analysis results.

• Klíčová slova
• IVF/ICSI, infertility, sperm analysis, sperm separation, spermatozoa,
• MeSH
• analýza spermatu metody   MeSH
• dospělí MeSH
• fragmentace DNA MeSH
• intracytoplazmatické injekce spermie * metody   MeSH
• lidé MeSH
• membránový potenciál mitochondrií MeSH
• mikrofluidika * metody   MeSH
• motilita spermií * MeSH
• mužská infertilita terapie   MeSH
• separace buněk * metody   MeSH
• spermie * cytologie   metabolismus   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

BACKGROUND: Given the lack of information concerning the role of Chlamydia trachomatis in male fertility, the aim of this study was to ascertain and analyze the quality of Chlamydiae-positive and -negative semen. MATERIAL AND METHODS: Sperm count was performed according to the 1999 World Health Organization (WHO) laboratory manual for examination of human semen and sperm-cervical mucus interaction, and sperm survival was assessed by a 120-min test. The evaluation of the morphological examination of ejaculates was carried out using the sasmo (strict morphological analysis of ejaculates) computer program. Chlamydiae were detected by immunofluorescent reaction using the Progen Biotechnik GmbH diagnostic set. Fisher's exact test and the chi-quadrate test were used for statistical analysis. RESULTS: Of the total of 627 sperm samples examined, Chlamydiae were detected in 136 cases (21.7%). Sperm analysis showed significant differences between Chlamydiae-positive and -negative samples. The Chlamydiae-contaminated group showed normal sperm morphology 14.4% lower, volume 6.4% lower, concentration 8.3% lower, motility 7.8% and velocity 9.3% lower than in Chlamydiae-negative samples. The average values for normal spermatozoa and motility in the Chlamydiae-positive group were also significantly reduced. CONCLUSION: Chlamydia trachomatis was found to be a possible factor in sperm pathology. These results could help to elucidate the role of Chlamydia trachomatis in male infertility.

• MeSH
• Chlamydia trachomatis imunologie   izolace a purifikace   MeSH
• chlamydiové infekce komplikace   MeSH
• ejakulace MeSH
• fluorescenční protilátková technika přímá MeSH
• lidé MeSH
• motilita spermií MeSH
• mužská infertilita etiologie   mikrobiologie   MeSH
• počet spermií MeSH
• sperma mikrobiologie   MeSH
• spermie fyziologie   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The aim of this study was to compare the spermatozoa quality parameters in spermatozoa of RAS (Recirculating Aquaculture System; RAS group) cultured (commercial pellets) and natural condition cultured (WILD group) burbot Lota lota (live prey, Pseudorasbora parva). Seven of nine fish of the RAS group produced sperm, with sperm from only four of the fish having a motility of >5%. Sperm were collected from all nine fish of the WILD group, and sperm of six of the fish from the WILD group had motility of about 100% and three had sperm with 50% to 60% motility. Spermatozoa from the RAS group had a delay in activation compared to the WILD group. Fish from the RAS group also had a lesser volume of sperm (1.8 ± 1.2 mL) collected compared to the WILD group (3.6 ± 1.2 mL). Compared to the RAS group, sperm of the WILD group had a greater proportion of saturated fatty acids (SFA), as well as the phospholipid, phosphatidylethanolamine. The findings indicate that fish grown in natural conditions may be more suitable as broodstock. Ongoing research to develop methods of enhancing reproductive performance of burbot broodstock cultured in RAS is needed to investigate whether the quality of sperm can be improved by adjusting environmental conditions, diet, or combination of these factors.

• Klíčová slova
• CASA, Milt, Reproduction, VCL, WOB,
• MeSH
• analýza spermatu * MeSH
• divoká zvířata MeSH
• Gadiformes metabolismus   fyziologie   MeSH
• lipidy analýza   MeSH
• mastné kyseliny analýza   metabolismus   MeSH
• metabolismus lipidů * MeSH
• spermie chemie   metabolismus   MeSH
• vodní hospodářství metody   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH
• Názvy látek
• lipidy MeSH
• mastné kyseliny MeSH

Accurate predictive modelling facilitates efficient and effective trait selection in animal breeding and can decrease costs while maximizing profits when raising economically important animals. The objective of this study was to extend a previously developed bioeconomic model and computer program to calculate the marginal economic values (MEVs) and economic weights (EWs) for direct and maternal pig traits affected by new reproductive traits, namely the number of sow functional teats (NFTs) and boar sperm quality traits (SQTs) that included sperm volume, sperm concentration, motility percentage and percentage of abnormal spermatozoa. The MEV of NFTs represented the cost differences between naturally and artificially reared piglets until weaning and the cost differences between naturally and artificially reared finished animals. The MEVs of SQTs expressed the saved costs for artificial insemination, assuming a decreased price per insemination dose when improving the SQTs. The absolute and relative EWs for the newly defined complex of traits in the breeding objectives for pig breeds involved in the Czech national three-way crossing system (Czech Large White [CLW], Czech Landrace [CL] and Pietrain [PN]) were calculated using gene flow methods. The NFT trait was included only for dam breeds, and the relative EW averaged 3.6% of the total economic importance based on the genetic standard deviations of all 19 simultaneously evaluated traits in CLW and CL breeds. The relative EWs of the four SQTs comprised 2.0% of the total economic importance of the 19 traits in the CLW and CL dam breeds and 8% of the total economic importance of the 18 traits in the PN sire breed. Therefore, inclusion of the NFTs for dam breeds and SQTs for sire breeds in the breeding goal is recommended to aid in obtaining ideal outcomes with optimal economic values.

• Klíčová slova
• crossbreeding, economic weights, number of functional teats, pigs, sperm quality traits,
• MeSH
• chov zvířat ekonomika   MeSH
• chov ekonomika   MeSH
• ekonomické modely MeSH
• fenotyp * MeSH
• mléčné žlázy zvířat * fyziologie   MeSH
• prasata genetika   fyziologie   MeSH
• rozmnožování genetika   MeSH
• selekce (genetika) MeSH
• spermie * fyziologie   MeSH
• tok genů MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH