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This study was designed to assess the dose- and time-dependent in vitro effects of α-tocopherol (α-TOC) on bovine spermatozoa during different time periods (Times 0h, 2h, 6h, 12h, 18h, 24h). Semen samples were collected from 20 adult breeding bulls, and diluted in physiological saline solution containing 0.5% ethanol (96%) together with 0, 1, 5, 10, 50, 100, 200, 500, 1000 and 2000 μM/L of α-TOC. Most α-TOC concentrations had beneficial effects on the spermatozoa motion parameters, while groups supplemented with 500 and 200 μM/L exhibiting significant differences (P

The general objective of this in vitro study was at first to evaluate copper (Cu) content in the cell sediment and seminal plasma fraction and compare their relationship with basic motility characteristics and secondly expand the knowledge of its impact on the fertilization potential of the spermatozoa. Semen samples were collected from 12 breeding bulls. The motility analysis was carried out using the Computer Assisted Semen Analysis (CASA) system. The mean value for the percentage of motile spermatozoa (MOT) was 91.24��5.17% and the progressive motility of the spermatozoa (PROG) as 88.58��5.85%. Subsequently, the samples were centrifuged to obtain the seminal plasma fraction and cell sediment. The seminal plasma Cu concentrations were analyzed by UV/VIS spectrophotometry. The total Cu concentration of the seminal plasma was 0.23 ��g/mL. The analysis by the flame atomic absorption spectrophotometry (FAAS) showed that the average cell sediment Cu concentration was 0.0014 ��g/mL. The correlation analysis revealed a moderate positive correlations between MOT and seminal plasma Cu concentration (rp=0.504; P>0.05) as well as between PROG and Cu content in the seminal plasma (rp=0.410; P>0.05). Copper in the cell sediment positively affected both MOT (rp=0.265) and PROG (rp=0.227), however, no significant differences were found (P>0.05). Conclusions of this study clearly indicated that Cu is important for the preservation of spermatozoa motility. The obtained results proved higher level of Cu in the seminal plasma, which seems representative of cumulative exposures of this trace element. Based on these results, we can conclude that the evaluation of total content of Cu in the whole semen is an important factor for determining the fertilization potential of spermatozoa., {"references":["AGARWAL, K., SHARMA, A., TALUKDER, G.: Clastogenic effects of copper sulphate on the bone marrow chromosomes of mice in vivo. 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The aim of this study was to analyze the influence of bovine serum albumin (BSA) used in different culture media on motility and viability of bovine spermatozoa during a 24-hour in vitro cultivation. We compared native samples (N) and samples cultivated in a commercial egg yolk medium (R) with samples cultivated in several experimental media containing 10% or 20% BSA. The analysis was carried out during three time periods (time 0, time 1 hour and time 24 hours) using Sperm VisionTM CASA system. Our study shows an obvious time-dependent decrease of the spermatozoa viability parameters in all experimental groups cultured for 24 hours. The lowest spermatozoa motility parameters were detected in native sample. Culture media with 10% BSA content showed only average quality and unsatisfactory results. Overall best motility parameters were detected when applying 20% BSA, especially in the medium D containig triladyl, 20% BSA and 5% glucose. Comparing results detected in this group with the results from the commercial egg yolk medium significant (P that BSA could be a good protein suplement for long-term bovine spermatozoa cultivation and further processing.

The target of this in vitro study was to analyse the influence of copper on the spermatozoa motility in the presence of bovine serum albumin (BSA) as culture medium and to provide additional information on the interaction between serum albumin and copper (II) chloride (CuCl2). The spermatozoa motility parameters was determined after exposure of CuCl2 (3.9; 7.8; 15.6; 31.2; 62.5; 125; 250; 500; 1000 μmol/L) using the Sperm VisionTMCASA (Computer Assisted Semen Analyzer) system during different time periods (Time 0 h, 1 h, 2 h and 24). The culture medium containing 20% BSA, triladyl and 5% glucose increased the overall percentage of spermatozoa motility after 1 h of cultivation. The percentage of motility spermatozoa significantly (P2 in comparison with the control group (without CuCl2administration). The experimental administration at the doses ≤ 31.2 μmol/L of CuCl2 stimulated the overall of motile spermatozoa (Time 24 h). Identical spermatozoa motility was detected also for the percentage of progressive motile spermatozoa during all time periods. Parameter of distance average path (DAP) showed increase in all CuCl2 addition groups in comparison with the control group at Time 1 h. Concentration 125 μmol/L of CuCl2 in various time periods of cultivation act stimulating on spermatozoa motility, but later (Time 24 h) inhibitory. Evaluation of velocity average path, showed similar results as for DAP. Measurement of the amplitude of lateral displacement (ALH) at Time 0 h as well as at Time 1 h was higher in all the experimental groups compared to the control group, but the differences were not significant (P>0.05). The experimental administration at the doses ≤ 62.5 μmol/L of CuCl2 stimulated ALH during 24 h of cultivation. The results suggest that adding energyand protein substrate to the culture medium increases the spermatozoa motility parameters also the presence of high doses (≥ 125 μmol/L) of copper ions during short-term periods (Time 0 h, 1 h). Concurrently BSA maintained motility of spermatozoa (≤ 31.20 μmol/L of CuCl2) during the long-term (Time 24 h) of cultivation, which confirms the protective effect of albumin binding to the copper ions.

The aim of this study was to examine the effects of glutathione, trehalose and caffeine on selected bovine spermatozoa morphology parameters, as well as lipid peroxidation following an in vitro cultivation at different temperatures and time periods. Morphological analysis showed similar occurrence of morphological abnormalities in all group with no significant differences (p>0.05). The TBARS assay revealed that the selected concentrations of trehalose, caffeine and glutathione did not have a significant effect (p>0.05) on lipid peroxidation as a process leading to oxidative stress development in spermatozoa.

Fertility is a complex process, involving multiple interactions between the sperm cells and seminal plasma components. The composition of seminal plasma has great influence on the biological quality of semen, as expressed by sperm motility and viability. The aim of this study was to evaluate relationships between semen characteristics and major nitrogen biochemical components of bovine seminal plasma with antioxidant properties (total proteins, albumin, uric acid, urea). Semen samples from 30 breeding bulls were examined. First, basic measurements were performed – volume (ml), pH and concentration (x 106/mL). Motility analysis was carried out using the CASA system; total protein, albumin, uric acid and urea concentration measurements were based on a colorimetric reaction of the target substance and a spectrophotometric detection at a specific wavelength. The correlation analysis revealed that the spermatozoa motility was positively correlated with total proteins (r=0.5823; p0.05) were found in relation to urea. This study proves the importance of proteins, albumin and uric acid in the semen quality, as well as the crucial role of biochemical analysis of semen in relation to male fertility.

Results of this study show that trehalose, caffeine and glutathione supplementations have stimulating effects on spermatozoa motility parameters. Stimulating effects of trehalose were lasting over a longer time period, with recommendation of trehalose supplementation for a long-time spermatozoa cultivation. Caffeine and glutathione have the most markedly stimulating effects especially over the first 24 hours of cultivation, and they could be used as additives for a short-term in vitro spermatozoa preservation.