Tomáš Slanina

Caffeine strongly improves motility parameters of turkey spermatozoa with no effect on cell viability

The purpose of this study was to evaluate the impact of caffeine on turkey spermatozoa during in vitro incubation. Experimental samples were prepared by diluting the raw semen with nine different concentrations of caffeine - from 0.078125 mg/mL to 10 mg/mL. The individual motility parameters were evaluated by the Computer Assisted Semen Analyser (CASA) system, and the viability of spermatozoa was evaluated using eosin-nigrosin staining. Selected parameters were recorded at six time periods: 0, 1, 2, 3, 4 and 5 h at 5 °C and 41 °C. A significantly higher motility and progressive motility of spermatozoa (P < 0.01 and P < 0.001, respectively) was detected in the samples containing caffeine ranging from 0.15625 to 7.5 mg/mL as compared to the control sample at 5 °C. At an incubation temperature of 41 °C the positive effect of caffeine on motility parameters was observed only at the beginning of incubation (at times 0 and 1). The tested caffeine concentrations showed no significant effect on the viability of turkey spermatozoa at any time period of incubation. A higher percentage of dead spermatozoa was observed for incubation at 41 °C (from 5.96% to 11.1%) in comparison to 5 °C (from 1.62% to 5.79%). The results suggest that caffeine can be used as a suitable component of turkey semen extenders and has the potential to improve fertility.

Lead induced alterations in rabbit spermatozoa motility and morphology in vitro

The aim of this in vitro study was to determine the effect of lead chloride (PbCl2) on rabbit spermatozoa motility and morphology. Lead concentrations in the medium ranged between 0.45 and 11.17 μg/ml; incubation time was 240 min (analyzed immediately after Pb addition followed by 30, 60, 120, 180, and 240 min), and temperatures of the culture environment were 22°C (laboratory), 4°C (refrigerator), and 37°C (incubator). Results were compared with a control group without Pb addition. After 30 min of culture at 22°C, a negative effect of Pb was noted as spermatozoa motility significantly decreased in groups with higher concentrations. After 120 and 240 min, a dose-dependent effect on spermatozoa motility was noted. At 4°C, spermatozoa motility analysis detected no significant differences between any of the experimental groups and control. At 37°C, a negative effect of Pb incubation on motility was detected at Times 30, 60, 120, 180, and 240 in groups with high concentrations. At Times 120, 180, and 240 a significant decrease in spermatozoa motility was also noted in all experimental groups in comparison to control. The analysis of pathological spermatozoa at Time 240 revealed an increasing trend of morphological abnormalities after incubation with Pb. Across three temperature regimes an increase of morphological changes was noted, particularly in the group with the highest Pb concentration. The predominant morphological abnormalities were knob twisted flagellum, flagellum ball, separated flagellum, and broken flagellum. Knob twisted flagella represented the most frequent pathological changes in the experimental group with the highest Pb concentration. Results suggest that the inhibitory effect of Pb on spermatozoa motility parameters depends on the concentration, incubation time, as well as environmental temperature during incubation. Furthermore, a negative effect of Pb in vitro on spermatozoa morphology indicates possible reproductive problems under in vivo conditions, too.

The effect of diluent, temperature and age on turkey spermatozoa motility in vitro

The aim of this study was to evaluate the turkey spermatozoa motility in in vitro conditions and to prove the effect of different conditions of incubation – diluents, temperature and age of birds. Spermatozoa were obtained from adult turkeys line of Big 6, and spermatozoa motility parameters were evaluated using a computer-assisted semen analyzer (CASA) system. Significant decrease of spermatozoa motility at laboratory temperature (22°C) was detected from time 0 (94.15%) till 180 minutes of incubation (53.91%). At the cool media incubation (5°C), this difference was lower (95.41 and 78.86%, respectively), and the differences were significant from 30 minutes of incubation till 180 minutes. Progressive spermatozoa motility replicated the tendency of total spermatozoa motility. When the physiological solution to commercial diluent at 5°C was compared, the spermatozoa motility and progressive motility in both groups were very consistent for 90 minutes of incubation. Subsequently, significantly higher spermatozoa motility was detected at time periods 120, 150 and 180 minutes of incubation in commercial diluent. Motility was also higher in this group after 24 hours. Influence of age on spermatozoa motility parameters was analysed at 22°C at the time 0 and after 30 minutes of incubation. Analysis of spermatozoa motility as well as progressive spermatozoa motility proved higher values in Group A (aged 35–42 weeks) compared to Group B (aged 63–73 weeks). These results clearly suggest that low temperature and commercial diluents maintain motility parameters during longer time periods and the increasing age of birds has negative impact on motility parameters.

Trace elements content in semen and their interactions with sperm quality and RedOx status in freshwater fish Cyprinus carpio: A correlation study

Objective of the present study was to investigate interactions between trace elements content and RedOx status, as well as sperm quality parameters (motility features, DNA fragmentation) in fish spermatozoa in natural conditions. Reproductively mature male freshwater fish (n = 16) of Cyprinus carpio breed were used in the study. Trace elements content was determined in fish milt samples by inductively-coupled plasma optical emission spectrometry (ICP-OES) and by cold-vapor atomic absorption spectroscopy (CV-AAS). Sperm quality evaluation was realized by computer-assisted sperm analysis (CASA) quantifying several parameters: concentration, total motility, progressive motility, distance average path, distance curved line, distance straight line, velocity average path, velocity curved line, velocity straight line, straightness, linearity, amplitude of lateral head displacement and beat cross frequency. The general scheme of descending concentrations of trace metals in semen samples was following: Zn > Fe > Cu > As > Sr > Ni > Mn > Se > Pb > Cr > Cd > Hg. Total motility of spermatozoa was relatively high (91.45%), however progressive motility was not even half of this value (39.47%). Sperm DNA fragmentation values were relatively low (4.00-6.29%). The percentage of immotile spermatozoa showed a significant correlation with all RedOx status parameters and also with DNA fragmentation. Positive statistically significant correlations were observed between trace elements (Mn, Se, Sr, and Zn) and some qualitative spermatozoa parameters (velocity and distance parameters). Cu and Hg content shows similar negative associations with progressive motility. Hg also interacted with production of malondialdehyde. Overall, the present study suggests application of multi-component mixtures of environmentally related trace elements concentrations when assessing the potential reproductive risk.