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Background: Conservation of species diversity is the need of the hour for preserving life forms on Earth. Extinction of any part of the ecosystem has negative impacts on many processes and systems. The objective of this work was to analyze some biochemical and molecular indicators and their correlations to biogenic elements and heavy metals in Testudo hermanni (n = 16). Methods: Biochemical parameters were analyzed using the commercial kit DiaSys and biochemical analyzer Randox RX Monza. Sodium, potassium, and chlorides were measured using the EasyLite analyzer. Oxidative stress was evaluated using colorimetric and luminometric methods. Quantification of chemical elements in the blood was carried out using inductively coupled plasma mass spectrometry (ICPS). Results: Biochemical values of analyzed samples from Hermann’s tortoises were almost the same as referential values described by multiple authors, with minor aberrations in the total protein parameter. Values of arsenic (As) and nickel (Ni) showed correlation with biochemical parameters and the parameters of oxidative stress. Cadmium (Cd) exhibited correlation with aspartate aminotransferase (AST). Conclusions: This study reports correlations among four heavy metals, and their levels were again correlated with biochemical and molecular parameters in Hermann’s tortoises.

The purpose of this study was to monitor concentration of selected biological and risk elements in testes and later epididymal spermatozoa motility of wild boars (Sus scrofa scrofa) as well as their association. Wild boars were hunted in region Žuhračka - Levice branch plant, Slovak Republic. Testes were surgically removed post-mortem and were transported to the laboratory. Spermatozoa were obtained by dissecting the distal portion of the cauda epididymis and were analysed by Computer assisted semen analyzer (CASA) system. Concentration of elements were measured by inductively-coupled plasma optical emission spectrometry (ICP-OES) and by cold vapor absorption spectrometer (MA-3 Solo Mercury Analyzer). Total spermatozoa motility was at the level of 44.29% and progressive motility was 18.47%. Concentration of elements in testes was in following order: K > Na > Mg > Ca > Fe > Zn > Al > Cu > Se > Mn > As > Cr > Pb > Mo > Sr > Ni > Ba > Cd > Li > Hg. Negative association was observed between Se and motility, progressive motility, further between As and velocity curved line and beat cross frequency. Mercury showed positive correlation with beat cross frequency. Our results suggest that some chemical elements accumulated from polluted environment can affect reproduction of wild animals.

Verbascoside is a compound that belongs to a group of natural substances present in plants used in traditional medicine. It possesses multiple biological capabilities including anti-inflammatory, anti-bacterial and anti-tumour properties. Aim of the study was to evaluate the effect of verbascoside in two different concentrations (1 g/kg – LVB and 2 mg/kg – HVB in food mixture) on selected parameters of mineral profiles in rabbit blood. In our study the experiment lasted for 120 days and was executed on 45 New Zealand rabbit bucks. The average age of animals was 1 year ± 2 month. Rabbits were divided into three homogenous groups – first being the control group fed with a feed without any additives (control); LVB group with 5 mg/kg and HVB group supplemented with 10 mg/kg of verbascoside. In our experiment verbascoside did not cause significant changes in parameters of mineral profile in rabbit blood. Levels of sodium in control group were 135.22 mmol/l. The levels of Na in LVB reached 135.64 mmol/l and in HVB group 134.74 mmol/l. Potassium analysis shown concentrations in the range of 4.32 mmol/l – 4.68 mmol/l with the lowest concentration in control group. The concentrations of Ca have remained almost unchanged compared to the control group with LVB group having the same concentration of 3.49 mmol/l and HVB group – 3.48 mmol/l. Chlorides concentrations ranged from 106.05 mmol/l to 160.61 mmol/l and did not show any significant changes. Magnesium blood levels in our experiment did not show any significant changes either and ranged from 1.32 mmol/l to 1.34 mmol/l. We can state that verbascoside had no effect on mineral profile parameters in rabbit blood, but it has to be emphasized that its effect is more focused on different functions in various targeted organs as stated above.

The aim of this study was to observe the effect of age on selected biochemical parameters of horses’ blood. Horses were divided in four age categories (5, 6, 7 and 9 years old). Blood was collected from 31 horses (Furioso breed). Assessment of biochemical parameters was realized by Randox RX Monza analyzer. Significant differences were observed between different groups in total proteins. Concentrations of calcium, phosphorus, sodium, potassium and chlorides correlate with the age of the animals. Similarly, liver enzyme activities correspond to age groups. In the other monitored biochemical parameters any significant differences between the groups were found. The achieved results confirm the connection between age of horses and values of biochemical parameters.

Livestock breeding has been carried out for several millennia based on an external assessment, i.e. exterior. Especially in horses that are bred for muscular work, the relationship between the shape of the body structure on the one hand and the purpose of use or performance on the other hand is very clear. At first glance, it is obvious that good horses in pull, jump or racing have different body shapes and not every formation of a body part provides a precondition for maximum performance. This knowledge has been known since the Middle Ages and led to the search for the ideal, the most beautiful or best animal. The performance of a sport and racing horse is conditioned not only by the quality of its gene pool, but also by many external factors. Of these, nutrition, the quality of breeding, the level of zootechnical care and, finally, the quality of the sports rider and coach dominate. An important factor is the process of adaptation to the load. The use of knowledge from the physiology of horse adaptation is the first step to intensify and improve the training process. The correct composition of the degree of load induces a whole complex of positive adaptive changes in trained individuals. Load limits must be set in such a way as to maintain all animal health requirements while achieving a high level of performance.

Article Details: Received: 2020-10-20 | Accepted: 2020-11-27 | Available online: 2021-01-31 https://doi.org/10.15414/afz.2021.24.mi-prap.1-4 Abstract The aim of the study was to observe the salivary concentrations of cortisol in horses during daily routine over the course of four weeks lasting experiment in three different stages of day to monitor the changing values of cortisol in horse saliva during the day. Saliva was obtained from 12 Slovak warmblood horses – 1 stallion, 6 mares and 5 geldings. In the experiment we focused purely on changes of salivary cortisol concentrations over the course of the daily routine.The results shown us multiple significant changes between individual sample collections and thus, we can state that according to our results the highest concentration of cortisol in horse saliva is in the morning and it decreases throughout the day with lowest measured concentrations being in the final sample collection of the day (at 22:00). These changes had no visible effect on horses´ organism throughout entirety of the experiment and were caused due to horses daily rhythm. Keywords: horse, cortisol, saliva, rest, daily routine References Bohak, Zs, Szabo, F, Beckers, J-F, Melo de Sousa, N, Kutasi, O., Nagy, K, Szenci, O. 2013. Monitoring the circadian rhythm of serum and salivary cortisol concentrations in the horse . Domest. Anim. Endocrinol. 45:38-42. https://doi.org/10.1016/j.domaniend.2013.04.001 Contreras-Aguilar, MD, Lamy, E, Escribano, D, Ceron, JJ, Tecles, F, Quiles, AJ, Hevia, ML. 2020. Changes in salivary analytes of horses due to circadian rhythm and season: A Pilot Study. Animals. 10(9):1486. https://doi.org/10.3390/ani10091486 Halo, M, Hollý, A, Mlynekova, E, Polyakova, L, Horný, M, Kovalcik, E. 2009. Influence feeding and training on the metabolic profil sport horses . J Cent. Eur. Agric. 10(4):411-416. Halo, M, Strapak, P, Mlynekova, E, Kovalcik, E, Horný, M. 2008. Influence stres on the training process of the horses. J. Cent. Eur. Agric. 9(1):217-224. Ille, N, von Lewinski, M, Erber, M, Wulf, M, Aurich, J, Mostl, E, Aurich, C. 2013. Effects of the level of experience of horses and their riders on cortisol release, heart rate and heart rate variability during a jumping course. Anim. Welf. 22:457-465. 10.7120/09627286.22.4.457 Irvine, CHG, Alexander, SL. 1994. Factors affecting the circadian rhythm in plasma cortisol concentrations in the horse. Domest. Anim. Endocrinol. 11(2):227-238. https://doi.org/10.1016/0739-7240(94)90030-2 Kang, O-D, Lee W-S. 2016. Changes in salivary cortisol concentration in horses during different types of exercise . Asian-Australas J Anim Sci. 29(5):747-752. 10.5713/ajas.16.0009 Kang, O-D, Yun Y-M. 2016. Influence of Horse and Rider on Stress during Horse-riding Lesson Program. Asian Australas J Anim Sci. 29(6):895-900. 10.5713/ajas.15.1068 Leal, BB, Alves, GES, Douglas, RH, Bringel, B, Young, RJ, Haddad, JPA, Viana, WS, Faleiros, RR. 2011. Cortisol circadian rhythm ratio: A simple method to detect stressed horses at higher risk of colic? Equine Vet. J. 31:188-190. https://doi.org/10.1016/j.jevs.2011.02.005 Massanyi, P, Stawarz, R, Halo, M, Formicki, G, Lukac, N, Cupka, P, Schwarcz, P, Kovacik, A, Tusimova, E, Kovacik, J. 2014. Blood concentration of copper, cadmium, zinc and lead in the horses and its relation to hematological and biochemical parameters. J. Environ. Sci. Health A. 49:973-979. 10.1080/10934529.2014.894322 Peeters, M, Sulon J, Beckers J-F, Ledoux D, Vandenheede M. 2011. Comparison between blood serum and salivary cortisol concentrations in horses using an adrenocorticotropic hormone challenge . Equine Vet J. 43(4):487-493. 10.1111/j.2042-3306.2010.00294.x Schmidt, A, Mostl, E, Wehnert, Ch, Aurich, J, Muller, J, Aurich, Ch. 2010. Corisol release and heart rate variability in horses during road transport . Hormones Behaviour. 57: 209-215. 10.1016/j.yhbeh.2009.11.003 Strzelec, K, Kankofer, M, Piertzak, S. 2011. Cortisol concentration in the saliva of horses subjected to different kinds of exercise. Act Vet. Brno. 80:101-105. 10.2754/avb201180010101 Stull, CL, Morrow, J, Aldridge, BA, Stott, JL, McGlone, JJ. 2008. Immunophysiological responses of horses to 12-hour rest during 24 hours of road transport. Vet. Rec. 162:609-614. 10.1136/vr.162.19.609 Van der Kolk, JH, Nachreiner, RF, Schott HC, Refsal KR, Zanella AJ. 2001. Salivary and plasma concentration of cortisol in normal horses and horses with Cushing´s disease. Equine Vet J. 33(2):211-213. 10.1111/j.2042-3306.2001.tb00604.x

Article Details: Received: 2020-09-29 | Accepted: 2020-11-27 | Available online: 2021-01-31 https://doi.org/10.15414/afz.2021.24.mi-prap.15-19 Over 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.1996.0292 Bucak, 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.2006.12.004 Erel, 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.2005.08.008 Halo, M. and Tirpak, F. (2018) Stallion fertility - the basis of successful reproduction. Svet koni . 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.x Reddy, 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-CJAS Stephens, 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.2012.10.004 Tirpak, 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.2017.7.2.128-131

Article Details: Received: 2020-10-21 | Accepted: 2020-11-27 | Available online: 2021-01-31 https://doi.org/10.15414/afz.2021.24.mi-prap.105-108 During their career, horses are affected by a large number of direct and indirect factors. The aim of our study was to determine the effect of indirect factors, such as year of start, sex, breed, number of starts, age and proportion of genes of Slovak Warmblood horse breed. For determination of indirect factors on performance level, results from test efficiency and final placement in showjumping category 4, 5 and 6 years old young horses were used. 540 horses of Slovak Warmblood breed were analysed between 2015 - 2019 and 1155 data samples were collected. Based on the results of the linear model, parameters – the effect of gender and the influence of the gene proportion of Slovak Warmblood horse had statistically significant influence (p

Feed additive consisting of polysaccharides from brown seaweeds plus phenolic acid, hydroxycinnamic acids, tannins, and flavonoids from plant extracts, was used as rabbit food supplement. Main aim of the study was to determine the effect of natural mix of marine and freshwater seaweed polyphenols on selected reproductive characteristics of male rabbits during the 90-days experiment. Natural mix was incorporated in feed-pellets for rabbits in two different concentrations – 0.3% (T1 group) and 0.6% (T2 group), compared with a control group (C group). In experimental groups a significant increase of concentration of calcium during first 30 days of supplementation was found. An increase of alanine aminotransferase, glutathione peroxidase and ferric reducing ability of plasma and a decrease in aspartate aminotransferase after 90 days were recorded in the same groups. Except for that we noticed decrease of semen distance of curved line and velocity of curved line after 30 days though only while being supplemented with 0.6% proportion of seaweed polyphenols in feed mixture. Based on the results it can be stated that the natural mix in the tested levels do not show adverse effect on male rabbit reproductive parameters, and an improvement of antioxidant status was observed. The feed additives can have a very important effect on growth, health and development of animals in general as it supplies the with the much-needed minerals, nutritional substances and antioxidants, on which we focused in our study.

Considering that spermatozoa are very sensitive cells, research of conservation media with different natural substances is very important area for many research institutions. Preservation media help to keep fertility ability of spermatozoa after long hours of storage and transport. In order to be able to use suitable natural substances as a conservation medium and successfully implement fertilization of a selected mare is necessary to ensure a thorough evaluation of spermatozoa. One of the most important evaluated parameters of stallion ejaculate quality is sperm concentration, motility and morphology. Commonly used methods for assessment of spermatozoa parameters are CASA system (Computer Assisted Semen Analysis) and mitochondrial toxicity test (MTT). This review summarises effect of taurine on stallion spermatozoa during storage and transport, especially in the development of potential parts conservation mediums.

Submitted 2020-07-02 | Accepted 2020-08-24 | Available 2020-12-01 https://doi.org/10.15414/afz.2020.23.mi-fpap.132-136 The present study aimed to analyse dried grape pomace as a possible source of crude fat and polyunsaturated fatty acids in equine nutrition, as well as its effect on apparent digestibility of crude fat and selected fatty acids. Twelve clinically healthy sport horses were used in the feeding trial (Slovak warm blood breed). Animals were divided into three groups; control group (without supplementation) and two experimental groups where diets were enriched by 200 g and 400 g of dried grape pomace, respectively. Digestibility analysis was carried out by total faeces collection method. Crude fat of feeds and faeces, extracted by Soxhlet-Henkel method, was subsequently subjected to fatty acid profile analysis by gas chromatography. Grape pomace contained 96.17 g.kg -1 of crude fat with linoleic (70.03% in fat) and oleic (15.86% in fat) as the most abundant fatty acids. An indication (P>0.05) of higher digestibility of crude fat and oleic acid in both experimental groups, in comparison with control group, was detected. The digestibility of palmitic, linoleic, α-linolenic and cis-11-eicosenoic acids was not affected by dried grape pomace consumption (P>0.05). Based on the results of this experiment, dried grape pomace had no significant effect neither on digestibility of crude fat nor on the selected fatty acids. However, this winery by-product could be used as an alternative source of crude fat in equine diets. Keywords: crude fat, equine, polyunsaturated fatty acid utilisation, wine by-products References Aslanian, A., Dizaji, A. A., Farhoomand, P., Shahryar, H. A., Sis, N. M., & Rouhnavaz, S. (2011). Characterization of the nutritive value and protein fractions the cornell net carbohydrate and protein system in White and Red Grape (Vitis vinifera sp.) Pomace. Research Journal of Biological Sciences, 6(7), 298-303. Azevedo, J. A. G., Valadares Filho, S. C., Pina, D. S., Detmann, E., Pereira, L. G. R., Valadares, R. F. D., ... & Benedeti, P. B. (2012). Nutritional diversity of agricultural and agro-industrial by-products for ruminant feeding. Arquivo Brasileiro de Medicina Veterinaria e Zootecnia, 64(5), 1246-1255. https://doi.org/10.1590/S0102-09352012000500024 Brenes, A., Viveros, A., Chamorro, S., & Arija, I. (2016). Use of polyphenol-rich grape by-products in monogastric nutrition. A review. Animal Feed Science and Technology, 211, 1-17. https://doi.org/10.1016/j.anifeedsci.2015.09.016 Burke, J. B. Equine International. (2009). Feeding Equine Athletes. Equine International, 1(2), 28-30. Davies, J. A., Krebs, G. L., Barnes, A., Pant, I., & McGrath, P. J. (2009). Feeding grape seed extract to horses: effects on health, intake and digestion. Animal, 3(3), 380-384. European Union. (2009). Commission regulation (EC) No 152/2009 of 27 Jan. 2009: Laying down the methods of sampling and analysis for the official control of feed. Foiklang, S., Wanapat, M., & Norrapoke, T. (2016). Effect of grape pomace powder, mangosteen peel powder and monensin on nutrient digestibility, rumen fermentation, nitrogen balance and microbial protein synthesis in dairy steers. Asian-Australasian Journal of Animal Sciences, 29(10), 1416-1423. https://doi.org/10.5713/ajas.15.0689 Galik, B., Kollathova, R., Rolinec, M., Juracek, M., Simko, M., Hanusovský, O., Biro, D., Vasekova, P., Kolesarova, A., Barantal, S. (2019). Grape by-products as bioactive substances in animal nutrition: A review. Agriculture and Food, 7, 67-172. Georgiev, V., Ananga, A., & Tsolova, V. (2014). Recent advances and uses of grape flavonoids as nutraceuticals. Nutrients, 6(1), 391-415. https://doi.org/10.3390/nu6010391 Gulcu, M., Uslu, N., Ozcan, M. M., Gokmen, F., Ozcan, M. M., Banjanin, T., … Lemiasheuski, V. (2019). The investigation of bioactive compounds of wine, grape juice and boiled grape juice wastes. Journal of Food Processing and Preservation, 43(1), e13850. https://doi.org/10.1111/jfpp.13850 Hanganu, A., Todasca, M. C., Chira, N. A., Maganu, M., & Rosca, S. (2012). The compositional characterisation of Romanian grape seed oils using spectroscopic methods. Food Chemistry, 134(4), 2453-2458. https://doi.org/10.1016/j.foodchem.2012.04.048 Hess, T., & Ross-Jones, T. (2014). Omega-3 fatty acid supplementation in horses. Revista Brasileira de Zootecnia, 43(12), 677-683. https://doi.org/10.1590/S1516-35982014001200008 Hinchcliff, K. W., Kaneps, A. J., & Geor, R. J. (2013). Equine Sports Medicine and Surgery E-Book. Elsevier Health Sciences. Hussein, S., & Abdrabba, S. (2015). Physico-chemical characteristics, fatty acid, composition of grape seed oil and phenolic compounds of whole seeds, seeds and leaves of red grape in Libya. International Journal of Applied Science and Mathematics, 2(5), 2394-2894. Kentucky Equine Research. (2016). Nutrition of the performance horse. Kollathova, R., Galik, B., Halo, M., Kovacik, A., Hanusovský, O., Biro, D., ... & Simko, M. (2020). The effects of dried grape pomace supplementation on biochemical blood serum indicators and digestibility of nutrients in horses. Czech Journal of Animal Science, 65(2), 58-65. https://doi.org/10.17221/181/2019-CJAS Lichovnikova, M., Kalhotka, L., Adam, V., Klejdus, B., & Anderle, V. (2015). The effects of red grape pomace inclusion in grower diet on amino acid digestibility, intestinal microflora, and sera and liver antioxidant activity in broilers. Turkish Journal of Veterinary and Animal Sciences, 39(4), 406-412. https://doi.org/10.3906/vet-1403-64 Mironeasa, S., Codina, G. G., & Mironeasa, C. (2016). The effects of wheat flour substitution with grape seed flour on the rheological parameters of the dough assessed by mixolab. Journal of Texture Studies, 43(1), 40–48. https://doi.org/10.1111/j.1745-4603.2011.00315.x National Research Council. (2007). National Research Council Committee nutrient requirements of horses. Piccione, G., Arfuso, F., Fazio, F., Bazzano, M., & Giannetto, C. (2014a). Serum lipid modification related to exercise and polyunsaturated fatty acid supplementation in jumpers and thoroughbred horses. Journal of Equine Veterinary Science, 34(10), 1181-1187. https://doi.org/10.1016/j.jevs.2014.07.005 Piccione, G., Marafioti, S., Giannetto, C., Panzera, M., & Fazio, F. (2014b). Effect of dietary supplementation with omega 3 on clotting time, fibrinogen concentration and platelet aggregation in the athletic horse. Livestock Science, 161, 109-113. https://doi.org/10.1016/j.livsci.2013.12.032 Ribeiro, L. F., Ribani, R. H., Francisco, T. M. G., Soares, A. A., Pontarolo, R., & Haminiuk, C. W. I. (2015). Profile of bioactive compounds from grape pomace (Vitis vinifera and Vitis labrusca) by spectrophotometric, chromatographic and spectral analyses. Journal of Chromatography B, 1007, 72–80. https://doi.org/10.1016/j.jchromb.2015.11.005 Ross-Jones, T., Hess, T., Rexford, J., Ahrens, N., Engle, T., & Hansen, D. K. (2014). Effects of omega-3 long chain polyunsaturated fatty acid supplementation on equine synovial fluid fatty acid composition and prostaglandin E2. Journal of Equine Veterinary Science, 34(6), 779-783. https://doi.org/10.1016/j.jevs.2014.01.014 Vineyard, K. R., Warren, L. K., & Kivipelto, J. (2010). Effect of dietary omega-3 fatty acid source on plasma and red blood cell membrane composition and immune function in yearling horses. Journal of Animal Science, 88(1), 248-257. https://doi.org/10.2527/jas.2009-2253 Viveros, A., Chamorro, S., Pizarro, M., Arija, I., Centeno, C., & Brenes, A. (2011). Effects of dietary polyphenol-rich grape products on intestinal microflora and gut morphology in broiler chicks. Poultry Science, 90(3), 566-578. https://doi.org/10.3382/ps.2010-00889

Nowadays great deal of research is physiological field is conducted on experimental animals and there is a lot of criticism from the wide public on methods used. Therefore, recently there is a lot of effort focused on the welfare of the animals. Main aim of this study is to determine the effect of experimental sample collection method on the selected parameters of stress. In the experiment two sample collections of rabbit blood from marginal ear vein were realized – first using standard method with one person fixing the animal and other collecting the blood using gently fixating the animal. In the second groups experimental method of inserting the experimental animal into a sack and further collection in dark was realized. During the experiment the levels of cortisol – main stress indicator in organism and other health parameters of animals including mineral profile and haematological parameters were observed. Our results show no significant changes in levels of cortisol but also a decreasing tendency in the sample from the second (dark) collection. Haematological parameters were generally in the reference values and any significant changes except levels of lymphocytes and percent of lymphocytes which shown significant increase in the second collection period were found. Also the levels of mean corpuscular haemoglobin and percent of neutrophils unveiled a significant decrease in values. Values of mineral profile parameters have indicated no significant changes except the levels of phosphorus. Based on the result we can state that the experimental sample collection has no effect on blood parameters of the animals but we spectated a statistically insignificant decrease in the levels of cortisol which can suggest that the dark collection is possibly less stressful to the animals.

The purpose of present study was to observe the effect of resorcinol on bovine spermatozoa before and after cryopreservation. Fresh semen was obtained from six randomly chosen breeding bulls. Experimental samples were prepared by diluting the semen with six different concentrations of resorcinol (REZ1– 4; REZ2 – 2; REZ3 – 1; REZ4 – 0.5; REZ5 – 0.25 and REZ6 –0.152 mg.ml-1). Experimental groups were compared against control group (REZK). Using CASA method spermatozoa motility was evaluated before cryopreservation and also after thawing, during incubation at 37°C in time periods 0, 5, 10, 15 and 60 minutes. Negative effect of in vitro resorcinol addition was observed in all analysed parameters (MOT, PRO, VCL, ALH and BCF) in groups REZ1, REZ2 and REZ3. In contrast, positive impact mainly on motility and progressive motility was found in experimental groups with the lowest resorcinol concentration addition (REZ4, REZ5 and REZ6) in comparison to the control group. After thawing significant effect of resorcinol on motility parameters were observed within each experimental group. Group with the highest concentration of resorcinol (REZ1) had markedly negative impact on evaluated motility parameters. Results of our study clearly confirm toxic effect of resorcinol on motility parameters of spermatozoa, which depend on concentration and time period.

The aim of this study was to compare the quality of musculus longissimus dorsi in the breeds of Mangalitsa and Large White with regard to the slaughter weight. Large White (LW) breed and White Mangalitsa (Ma) breed were used in the experiment. The system of housing and feeding was the same in both of the monitored breeds. The pigs were fed with the same feeding mixture ad libitum. According to the slaughter weight, the pigs were divided into three groups: up to 100 kg, 101 – 110 kg and over 110 kg. The breed Ma had a significantly lower drip loss than the breed LW. Evaluating the color of the meat, the LW breed has showed significantly higher L* (lightness, white ±black) and lower a* (redness, red ± green) values than the Ma breed. Within the chemical meat composition, the Ma breed had a significantly higher water content in MLD compared to the LW breed. Generally, there were no major differences in the meat quality between the Mangalitsa and Large White breeds. Finally it can be concluded that the breed Mangalitsa showed more favorable values of the physico-chemical indicators. Comparing the quality of the meat with regard to the slaughter weight, there were no large differences between individual weight groups. A higher slaughter weight has positively influenced mainly the color of the meat, as pigs weighing more than 110 kg achieved a significantly lower value of L* and a higher value of a* in comparison to pigs of the lower weight. As a positive effect of a higher slaughter weight can be considered its effect on the protein content in the meat, as pigs weighing over 100 kg have a significantly higher protein content in the meat than pigs weighing below 100 kg.

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.

Zinc plays a very important role in various biological activities of the body. Multifaceted role of zinc is also known in testes development, spermatogenesis, capacitation and has effect on spermatozoa motility. On the other hand, the growing industry of nanotechnology has created reasonable interest of the risk assessment for nanoparticles. The aim of this study was to evaluate in vitro effect of zinc oxide (ZnO) nanoparticles on rabbit spermatozoa. Fresh semen was collected from sexually mature New Zealand rabbits. Experimental groups were prepared by diluting semen with ZnO nanoparticles in seven different concentrations (6–391 mg/mL). The experimental groups were compared with control group. Semen was assessed using computer assisted semen analysis (CASA) at intervals of 0, 1, 2 and 3 h of incubation. The mitochondrial toxicity assay (MTT) assay was used to determine cell viability. The results of monitored motility parameters in experimental groups showed a decreasing trend during whole experiment. Significant decrease (P

The target of this study was to evaluate the effect of extract of the European mistletoe – Viscum album quercus L. on spermatozoa motility and viability in vitro. The CASA system was used to determine the spermatozoa motility parameters at different time intervals (0, 1, 2 and 3 h) and spermatozoa viability was determined in five different doses of Viscum album quercus L [10 (QA), 6.6 (QB), 3.3 (QC), 2.5 (QD) and 2 (QE) mg/ml]. Results in experimental groups detected a significant deterioration on rabbit spermatozoa after 1, 2 and 3 hours, compared to the control. The initial total spermatozoa motility showed increased value for all doses of Viscum album quercus in comparison to control. After in vitro culture a dose–dependent decrease (QA: reduction of 69.7 %, QB: reduction of 40.9 %) was found. For the progressive spermatozoa most significant decrease (86.8 % for QA vs. 48.5 % for QB) was detected compared to the control after 3 hours of culture. Spermatozoa viability (MTT test) was decreased in all experiment groups at the end of experiment, but the differences were not significant. Significant alterations of membrane integrity were found in groups with the highest Viscum album quercus concentration (QA, QB), but acrosome integrity showed no significant changes. Results suggest negative dose– and time–dependent effect of Viscum album quercus at higher doses on spermatozoa motility and viability parameters in vitro.

Mistletoe (Viscum album L.) was previously described as an effective additive in the therapy of patients with cancer after medical therapy of the primary neoplasia. The defence activity and also direct cytotoxic stimulation was found to be upstanding for the anti-carcinogenic action. The main target of this study was to detect the effect of Viscum album pini L. at various concentration 1.0 (PA); 0.66 (PB); 0.33 (PC); 0.1 (PD); 0.066 (PE) and 0.033 (PF) mg/mL on the fine rabbit spermatozoa motility parameters at various time periods (0; 1; 2 and 3 h) during the cultivation in vitro. The CASA methodology was applied to estimate the spermatozoa motility parameters. The spermatozoa motility at the beginning of the experiment (Time 0) found decreased rate for examined doses of Viscum album pini compared to the control, but the statistical significant reduction was observed (p

The objective of this study was to analyse the level of pedigree and genomic inbreeding in a herd of the Norik of Muran horses. The pedigree file included 1374 animals (603 stallions and 771 mares), while the reference population consisted of animals that were genotyped by using 70k SNP platform (n = 25). The trend of pedigree inbreeding was expressed as the probability that an animal has two identical alleles by descent according to classical formulas. The trend of genomic inbreeding was derived from the distribution of runs of homozygosity (ROHs) with various length in the genome based on the assumption that these regions reflect the autozygosity originated from past generations of ancestors. A maximum of 19 generations was found in pedigree file. As expected, the highest level of pedigree completeness was found in first five generations. Subsequent quality control of genomic data resulted in totally 54432 SNP markers covering 2.242 Mb of the autosomal genome. The pedigree analysis showed that in current generation can be expected the pedigree inbreeding at level 0.23% (ΔFPEDi = 0.19 ± 1.17%). Comparable results was obtained also by the genomic analysis, when the inbreeding in current generation reached level 0.11%. Thus, in term of genetic diversity both analyses reflected sufficient level of variability across analysed population of Norik of Muran horses.

The objective of this study was to estimate the distribution of autozygosity islands represented by homozygous segments (ROHs) in the genome of the Slovak Warmblood horse. The Slovak Warmblood is a very efficient breed with the excellent characteristics of a sport horse. The study included 37 animals that were genotyped by GGP Equine70k chip (71,947 SNPs). Only animals (36) and autosomal SNPs (62,439) with call rate >90% and minor allele frequency >1% were included in subsequent analyses. The homozygous segments were defined as stretches with minimum 15 consecutive homozygous SNPs of >500 kb with minimum density 1 SNP per 100 kb and maximum gap between markers of 1,000 kb. The heterozygous or missing calls were not accepted. The analysis indicated in total of 8,501 autozygosity islands in the genome of the Slovak Warmblood horse. The majority of identified segments (85.42%) were most likely derived from the remote ancestors in the past. Only 0.15% of detected segments resulted from the recent selection events affecting the genetic structure of studied population. The proportion of segments varied across chromosomes. The major fraction of autosome residing in ROH was found on ECA1 (8.30%), while ECA31 showed the lowest ROH coverage. The scan for overlapping homozygous segments shared by more than 50 % of animals demonstrated that the ECA6 autosome may be under strong selection pressure. Inside those selection signals, several genes were identified including them associated with immunity and reproduction.

Zinc is an essential trace element, which plays important roles in different biological activities. The aim of this study was to evaluate the effect of zinc on rabbit spermatozoa during in vitro conditions. Fresh semen was obtained from 8 sexually mature New Zealand White rabbits, which were bred at National Agricultural and Food Centre in Lužianky. Experimental groups were prepared by diluting the semen with two forms of zinc (ZnCl2, ZnSO4 .7 H2O) in five different concentration: ZA, ZA1 (4.883 mg Zn.l-1), ZB, ZB1 (9.766 mg Zn.l-1), ZC, ZC1 (19.531 mg Zn.l-1), ZD, ZD1 (39.063 mg Zn.l-1), ZE, ZE1 (78.125 mg Zn.l-1). Experimental samples were compared against control groups (ZK, ZK1). Semen was evaluated using the Computer Assisted Semen Analysis (CASA) at time intervals 0, 60, 120 and 180 minutes after incubation at 39°C. MTT test was used for determination of cell viability. Results of monitored motility parameters (motility, progressive motility and velocity curved line) showed decreasing trend in experimental samples. Significantly lower values of motility were observed in sample ZE, where percentage of motility decreased to 9.18±1.34% and after 180 minutes of incubation. Evaluation of spermatozoa velocity curved line detected significant (P

The use of artificial insemination in cattle breeding has evolved to global extent, and insemination doses are often shipped via air transport which requires strict radiation-based examinations. For the determination of effect of non-ionizing radiation (NIR), to which are beings frequently exposed due to protection of airport or cultural event security, freshly ejaculated and cryopreserved bovine spermatozoa were used as experimental model. Following radiation with hand-held metal detector in various exposition times (0, 10 s, 15, 30 and 60 min-groups FR, FR10, FR15, FR30 and FR60) the spermatozoa underwent motility and DNA fragmentation analyses. Study on cryoconserved semen treated with NIR was performed in time intervals 0, 10 s, 1 and 5 min (insemination doses radiated before cryoconservation-CB, CB10, CB1, CB5; samples radiated after freezing-CA, CA10, CA1 and CA5). Fresh semen and insemination doses radiated after cryoconservation showed significantly lower total and progressive motility. No effect on motility parameters was detected in semen extended with cryopreservative medium and radiated prior to freezing. Surprisingly, NIR showed a potential to stimulate spermatozoa velocity; however, the effect was modulated throughout the post-thawing incubation. Based on the DNA fragmentation assay, sperm DNA stayed intact. Present study underlines the potential harm of NIR, which is frequently used in everyday life, with overall adverse impact on human and animal reproduction. Current study also points out on interesting short-term spermatozoa stimulation induced by NIR.

One of the important factors that significantly affects the intensity and content of training and determines the performance it the horse interior. Character and temperament traits are described in the breeding goals of each breed of horse. The aim of this study was to analyze the horse interior and comparison the interior characteristics of two studied horse breeds Slovak warmblood horses and Lipizzan horses. Into the analysis were included 65 horses from National Stud Farm Topoľčianky, Slovak warmblood horses (n1=33) and Lipizzan horses (n2=32). Interior was analyzed through a questionnaire with 13 indicators with 10 point system, where 10 being the highest rating. Slovak warmblood horses become the highest ratings (average 89.1 pt, 74.42%) and Lipizzan horses (86.1 pt, 66.2%), so both breeds of horses can be considered as the breeds with balanced characteristics of temperament and character. But Lipizzan horses are more balanced, because their results had lower variability. There were insignificant differences between the analyzed horses in the studied breeds. Nevertheless, a several analyzes of individual indicators showed that between the Slovak warmblood horses and Lipizzan horses there is a significant difference in a single indicator of interior - 10 - Stress managing. In particular, it has been shown that Lipizzan horses manage stress significantly better than Slovak warmblood horses., Jedným zdôležitých faktorov, ktorý podstatne ovplyvňuje intenzitu a náplň tréningu a udáva výkonnosť koní je interiér. Vlastnosti charakteru a temperamentu sú popísané v chovných cieľoch jednotlivých plemien koní. Cieľom našej práce bolo analyzovať interiér koňa a porovnať interiérové vlastnosti dvoch skúmaných plemien koní, slovenského teplokrvníka a lipicana. Do analýzy bolo zahrnutých 65 koní z Národného žrebčína Topoľčianky, slovenský teplokrvník (n1=33) a lipican (n2=32). Interiér sme analyzovali prostredníctvom dotazníka s 13 ukazovateľmi 10 bodovým systémom, kde 10 bola najvyššia známka. Kone plemena slovenský teplokrvník získali najvyššie hodnotenie (priemerne 89,1 bodov, 74,42%) a kone plemena lipican (86,1 bodov, 66,2%), takže obe plemená môžeme považovať za plemená s vyrovnanými vlastnosťami charakteru a temperamentu. Avšak kone plemena lipican sú viac vyrovnané, pretože ich výsledky majú nižšiu variabilitu. Zaznamenali sme štatisticky nevýznamné rozdiely medzi analyzovanými plemenami koní. Avšak niekoľko analýz jednotlivých ukazovateľov ukázalo, že medzi koňmi slovenského teplokrvníka a lipicana je štatisticky významný rozdiel v jedinom ukazovateli interiéru – 10 – Zvládanie stresu. Konkrétne sa ukázalo, že kone plemena lipican zvládajú stre s významne lepšie ako kone plemena slovenský teplokrvník.

The aim of this study was to determinate the effect of various taurine concentrations on rabbit spermatozoa in in vitro conditions with the potential use in routine artificial insemination. Taurine, amino acid distinguished by its strong antioxidant properties, has been widely tested on spermatozoa of all kinds of farm animals except for the rabbits. This study investigated differences in rabbit spermatozoa motility and viability between the control sample and samples enriched with taurine solutions: TAU3 (3.125 mM), TAU6 (6.250 mM) and TAU12 (12.5 mM). Ejaculates of nine adult rabbits were subjected to this study and observed at time intervals 0, 90, 180 and 270 minutes while cultivated at 39°C. For detection of viability the MTT test was used and motility parameters were carried out using CASA analysis. Experimental samples were compared against the control. Assessment of the mitochondrial toxicity revealed that the addition of taurine ensured the higher viability in supplemented samples however with no statistical significance. Monitored motility parameters (motility, progressive motility and velocity curved line of spermatozoa) showed that taurine treated spermatozoa have lower activity at the beginning of the study but along with increasing time the positive effect of taurine was induced. Significant (P