High taurine concentrations negatively effect stallion spermatozoa parameters in vitro

Article Details: Received: 2020-09-29 | Accepted: 2020-11-27 | Available online: 2021-01-31https://doi.org/10.15414/afz.2021.24.mi-prap.15-19Over the past decades natural substances are widely used in the maintaining of spermatozoa viability. The target of present study was to evaluate the effect of various taurine concentrations on stallion spermatozoa during 37°C cultivation. Fresh semen was collected from 10 breeding stallions. The experimental groups were supplemented with six different concentration of taurine (in mg/ml): A – 2.5, B – 5, C – 7.5, D – 10, E – 15, F – 20 and compared to control (CON – 0). Spermatozoa motility was assessed using the Computer Assisted Semen Analyzer (CASA) system in 6 time periods (0, 1, 2, 3, 4 and 5 hours). The MTT test was used for detection of viability. For measuring antioxidant activity FRAP and TOS methods were used. Significantly negative effect was observed in the samples with the highest concentration of taurine (20 mg/ml). Spermatozoa viability was not significantly affected in analysed concentrations of taurine. Significant higher antioxidant activity was detected in the sample with the highest taurine concentration. Data clearly showed negative effects of high taurine concentrations on stallion spermatozoa.Keywords: taurine, CASA, antioxidant activity, spermatozoa, stallion References  Benzie, I. F. and Strain, J. J. (1996). The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: The FRAP assay. Analytical Biochemistry, 239(1), 70-76. https://doi.org/10.1006/abio.1996.0292Bucak, M. N. et al. (2007). The influence of trehalose, taurine, cysteamine and hyaluronan on ram semen: Microscopic and oxidative stress parameters after freeze–thawing process. Theriogenology, 67(5), 1060-1067. https://doi.org/10.1016/j.theriogenology.2006.12.004Erel, O. (2005). A new automated colorimetric method for measuring total oxidant status. Clinical Biochemistry, 38(12), 1103-1111. https://doi.org/10.1016/j.clinbiochem.2005.08.008Halo, M. and Tirpák, F. (2018) Stallion fertility - the basis of successful reproduction. Svet koní. 18.Halo Jr., M. et al. (2019) Time and dose-dependent effects of Viscum album quercus on rabbit spermatozoa motility and viability in vitro. Physiological Research, 68(6), 955-972. https://doi.org/10.33549/physiolres.934223 Ijaz, A. and Ducharme, R. (1995). Effect of various extenders and taurine on survival of stallion sperm cooled to 5 C. Theriogenology, 44(7), 1039-1050. https://doi.org/10.1016/0093-691x(95)00290-o Jambor, T. et al. (2017) In vitro effect of 4-nonylphenol on human chorionic gonadotropin (hCG) stimulated hormone secretion, cell viability and reactive oxygen species generation in mice Leydig cells. Environmental Pollution, 222, 219–225. https://doi.org/10.1016/j.envpol.2016.12.053 O'flaherty, L. et al. (1997) Intestinal taurine transport: a review. European Journal of Clinical Investigation, 27(11), 873-880. https://doi.org/10.1046/j.1365-2362.1997.2000747.xReddy, N. S. S. et al. (2010). Effects of adding taurine and trehalose to a tris-based egg yolk extender on buffalo (Bubalus bubalis) sperm quality following cryopreservation. Animal Reproduction Science, 119(3-4), 183-190. https://doi.org/10.1016/j.anireprosci.2010.01.012 Slanina, T. et al. (2018) Effect of taurine on turkey (Meleagris gallopavo) spermatozoa viability and motility. Czech Journal of Animal Science, 63(4),127-135. https://doi.org/10.17221/79/2017-CJASStephens, T. D. et al. (2013) Effects of pentoxifylline, caffeine, and taurine on post-thaw motility and longevity of equine frozen semen. Journal of Equine Veterinary Science, 33(8), 615-621. https://doi.org/10.1016/j.jevs.2012.10.004Tirpák, F. et al. (2017) Low taurine concentrations possitively affect rabbit spermatozoa properties in later time intervals. Journal of Microbiology, Biotechnology and Food Sciences, 7, 128-131. https://doi.org/10.15414/jmbfs.2017.7.2.128-13

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

The effects of caffeine on the motility and viability of stallion spermatozoa at different temperature conditions

The purpose of this study was to evaluate the dose-and time-dependent effect of caffeine treatment on the motility and viability of stallion spermatozoa at different temperatures. Six dose groups (A to F) were established with changing caffeine concentrations (from 0.625 to 10 mg/mL). The control samples were prepared by diluting the ejaculate only with physiological salt solution. The samples were examined after 0, 1, 2 and 3 h of incubation at 5 degrees C and 37 degrees C. The motility parameters were evaluated by Computer Assisted Semen Analyzer (CASA) system, and the viability was assessed by the mito-chondrial toxicity test at the end of the incubation. A positive effect of the lowest tested caffeine concentration on the motility parameters was observed throughout the incubation period at 5 degrees C. At the end of the 3h incubation, the viability in every sample in these groups, treated with any caffeine concentration, showed lower values compared to the control. At the higher incubation temperature (37 degrees C), caffeine positively affected the motility in samples B (P < 0.05) and D, E, F (P < 0.001) after 3 h of incubation; however, the viability showed a slightly decreasing tendency. Our results suggest that caffeine, in an optimal concentration, may be used as a component of stallion semen extenders

Exogenous Factors Affecting the Functional Integrity of Male Reproduction

Natural processes along with increased industrial production and the irresponsible behavior of mankind have resulted in environmental pollution. Environmental pollutants can be categorized based on their characteristics and appearance into the following groups: physical, biological, and chemical. Every single one of them represents a serious threat to the male reproductive tract despite the different modes of action. Male gonads and gametes are especially vulnerable to the effect of exogenous factors; therefore, they are considered a reliable indicator of environmental pollution. The impact of xenobiotics or radiation leads to an irreversible impairment of fertility displayed by histological changes, modulated androgen production, or compromised spermatozoa (or germ cells) quality. The present article reviews the exogenous threats, male reproductive system, the mode of action, and overall impact on the reproductive health of humans and animals

A Combination of Taurine and Caffeine in Stallion Semen Extender Positively Affects the Spermatozoa Parameters

This study was aimed to determine the impact of different taurine and caffeine combinations on the motility, viability, and oxidative markers of chilled stallion spermatozoa. Each stallion semen sample was diluted in a ratio of 1:2, with various taurine and caffeine concentrations (2.5–7.5 mg/mL taurine + 0.625–1.25 mg/mL caffeine) dissolved in a conventional extender. The control samples (CON) were prepared by diluting ejaculate only using the conventional extender. The motility was analyzed using a CASA system at different time intervals (0, 6, 12, 24, and 30 h) and the viability was evaluated using a mitochondrial toxicity test (MTT) performed at the end of the incubation at 5 °C. The liquid part of experimental samples was separated by centrifugation after 30 h of incubation and underwent the evaluation of oxidative stress via the quantification of markers ferric reducing ability of plasma (FRAP) and total oxidant status (TOS). The samples that were treated with a combination of taurine and caffeine significantly improved the motility parameters, mainly after 12, 24, and 30 h of incubation. Samples extended with combination of taurine and caffeine neither compromise viability nor alterations of redox status. The results of this study describe the combination of taurine and caffeine as an optimal supplement for improving the quality of stallion semen during chilled storage

Sperm Quality Affected by Naturally Occurring Chemical Elements in Bull Seminal Plasma

This study monitored the chemical and biochemical composition of bovine seminal plasma (SP). Freshly ejaculated semen (n = 20) was aliquoted into two parts. The first aliquot was immediately assessed to determine the sperm motion parameters. Another motility measurement was performed following an hour-long co-incubation of spermatozoa with SP at 6 &deg;C. The other aliquot was processed to obtain the SP. Seminal plasma underwent the analyses of chemical composition and quantification of selected proteins, lipids and RedOx markers. Determined concentrations of observed parameters served as input data to correlation analyses where associations between micro and macro elements and RedOx markers were observed. Significant correlations of total oxidant status were found with the content of Cu and Mg. Further significant correlations of glutathione peroxidase were detected in relation to Fe and Hg. Furthermore, associations of chemical elements and RedOx markers and spermatozoa quality parameters were monitored. The most notable correlations indicate beneficial effects of seminal Fe on motility and Mg on velocity and viability of spermatozoa. On the contrary, negative correlations were registered between Zn and sperm velocity and seminal cholesterol content and motility. Our findings imply that seminal plasma has a prospective to be developed as the potential biomarker of bull reproductive health

Composition of Stallion Seminal Plasma and Its Impact on Oxidative Stress Markers and Spermatozoa Quality

The composition of seminal plasma of individual sires varies and so does the fertilizing ability. Micro and macro elements along with seminal enzymes, hormones, proteins, and lipids contained in seminal plasma are essential for the proper physiological function of spermatozoa. However, elevated levels against the normal physiological values, especially in the case of trace metals, result in the production of reactive oxygen species. The deficiency of antioxidants in the seminal plasma that could scavenge free radicals causes an impairment of spermatozoa quality. Ejaculates were obtained from 19 stallions. The fresh semen was analyzed to evaluate qualitative parameters of spermatozoa in terms of the motility, viability, and integrity of DNA. Separated seminal plasma underwent the assessment of the chemical and biochemical composition and RedOx markers. Based on the obtained concentrations of individual chemical elements, the correlation analysis suggested a negative impact of Cu in seminal plasma on the SOD, GPx, and LPO. Contrary, positive correlation was detected between FRAP and motility features. While Cu negatively correlated with sperm motion parameters, the adverse effect on viability was suggested for Cd. Our data suggest that seminal plasma has a potential due to its availability to become the potential biomarker of the reproductive health of farm animals