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1

Molecular forms of selected antioxidant enzymes in dog semen – electrophoretical identification

100%

Koziorowska-Gilun M. , Strzezek R.

Polish Journal of Veterinary Sciences

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2011

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tom 14

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nr 1

EN

The aim of the study was the electrophoretical identification of molecular forms of selected antioxidant enzymes in dog semen. Ejaculates to be studied were chosen from five dogs, aged from two to eight years. Polyacrylamide gel electrophoresis was carried out under non-denaturing conditions and then gels were stained for the activity of the following enzymes: superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx). Sperm homogenates and all fractions (pre-spermatic, spermatic and post-spermatic) of dog ejaculate demonstrated one protein band with SOD activity characterized by low electrophoretic mobility. Based on the confirmed sensitivity to H₂O₂, it can be assumed that the detected SOD is an enzyme containing ions of Zn²⁺ and Cu²⁺ (Cu,Zn SOD). In sperm homogenates one protein band with GPx activity was characterized by high electrophoretic mobility, whereas in the spermatic and post-spermatic fractions of dog ejaculate three protein bands with different (low, medium and high) electrophoretic mobility were identified. CAT molecular forms were not found in either sperm homogenates or in the analyzed fractions of ejaculate.

2

Effect of dialysis of dog semen on sperm characteristics and some biochemical components of seminal plasma

80%

Strzezek R. , Koziorowska-Gilun M. , Kielczewski K. , Kordan W.

Polish Journal of Veterinary Sciences

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2015

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tom 18

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nr 2

EN

The aim of this study was to investigate the effect of dog semen dialysis on sperm characteristics and some biochemical components of seminal plasma. Whole ejaculates were dialyzed against Triscitrate-fructose extender for a 5 h period at room temperature (using semi-permeable cellulose tubing of 12-14 kDa molecular weight cut-off). It has been demonstrated that the long-term dialysis of dog semen causes a significant decrease in sperm quality parameters and disrupts the biochemical properties of seminal plasma. This procedure requires further improvement.

3

Characteristics of antioxidant system in dog semen

80%

Strzezek R. , Koziorowska-Gilun M. , Kowalowka M. , Strzezek J.

Polish Journal of Veterinary Sciences

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2009

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tom 12

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nr 1

EN

The aim of the study was to characterize the enzymatic and non-enzymatic components comprising the antioxidant system in spermatozoa and individual fractions of dog ejaculate. Ejaculates were collected from six dogs of mixed-breeds. Total protein content, activity of antioxidant enzymes and the content of low-molecular antioxidants, such as L-glutathione (GSH), L-ergothioneine (ERG), L-ascorbic acid and total SH-group, were analyzed in the ejaculated spermatozoa and seminal plasma of the pre-spermatic, spermatic and post-spermatic fractions. The total antioxidant status (TAS) and antiperoxidant activity of the seminal plasma were also determined. The enzymatic antioxidant system of canine spermatozoa is mainly represented by superoxide dismutase (SOD) activity and, to a lesser extent, by glutathione peroxidase (GPx) and phospholipid hydroperoxide glutathione peroxidase (PHGPx) activity. Catalase activity was not detected either in the spermatozoa or in the different ejaculate fractions. GSH and ERG were detected in each fraction. Furthermore, a high level of L-ascorbic acid was observed in fractions of the ejaculate. Proteins and low-molecular weight antioxidants could influence the total antioxidant status and antiperoxidant activity of the seminal plasma. Increased suppressive activity against lipid peroxidation was shown only in the pre-spermatic and post-spermatic fractions. The different fractions of dog ejaculate, which are the main source of enzymatic antioxidants and low-molecular antioxidants, play an important role in protecting spermatozoa against reactive oxygen species.

4

Expression of hypoxia-inducible factor-1alfa and association with Cu/Zn superoxide dismutase in the reproductive tract tissues of adult male roe deer (Capreolus capreolus) during the reproduction season

80%

Koziorowska-Gilun M. , Gilun P. , Koziorowski M. , Kordan W.

Annals of Animal Science

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2017

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tom 17

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nr 4

5

Antioxidant status of spermatoza of cauda epididymis and fluids of accessory sex glads of boar

80%

Koziorowska-Gilun M. , Kowalowka M. , Koziorowski M. , Strzezek J.

Acta Biochimica Polonica

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2006

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tom 53

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nr Supplement 1

6

Seasonal changes in antioxidant defence systems in seminal plasma and fluids of the boar reproductive tract

80%

Koziorowska-Gilun M. , Koziorowski M. , Strzezek J. , Fraser L.

Reproductive Biology

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2011

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tom 11

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nr 1

7

Acrosin system of dog spermatozoa and reproductive tract secretions

80%

Strzezek R. , Koziorowska-Gilun M. , Filipowicz K.

Polish Journal of Veterinary Sciences

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2010

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tom 13

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nr 1

69-73

EN

The aim of this study was to determine the activity of proacrosin and acrosin in spermatozoa originating from the sperm-rich fractions (SRF) and whole ejaculates (WE) of dog semen. In addition, experiments were conducted to determine the activity of antitrypsin inhibitors in the fluids of different ejaculate fractions and whole seminal plasma. Ejaculates were collected from five dogs of mixed breed and one Beagle dog (aged from 2 to 9 years). In the SRF, it was confirmed that the activity of the free acrosin form was predominant (acrosin / proacrosin; 2.38 ± 0.22 / 1.05 ± 0.08 mIU / 106 spermatozoa). On the other hand, spermatozoa originating from the WE exhibited significantly higher (p<0.05) proacrosin activity (proacrosin / acrosin; 2.19 ± 0.19 /1.30 ± 0.11 mIU / 106 spermatozoa). Furthermore, acrosin inhibitor activity was lower in the fluids of the pre-sperm fraction (0.09 ± 0.006 IU / cm3), whereas it was higher in the fluids of the post-sperm fraction (0.11 ± 0.007 IU / cm3). Using PAGE analysis, the antitrypsin activity of the enzyme was represented by the presence of one electrophoretic band in the fluids of the pre-sperm and post-sperm fractions and whole seminal plasma. Furthermore, two electrophoretic bands were detected in the fluids of the SRF. The findings of this study indicate that specific proteinase inhibitors present in the individual ejaculate fractions of dog semen may act by stabilizing the sperm acrosin system.

8

Cryopreservation of canine semen: the effect of two extender variants on the quality and antioxidant properties of spermatozoa

80%

Strzezek R. , Koziorowska-Gilun M. , Stawiszynska M.

Polish Journal of Veterinary Sciences

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2012

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tom 15

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nr 4

EN

The aim of this study was to determine the effect of two variants of Tris-citric acid-fructose (TCF) extender containing whole hen egg yolk (TCF-HEY) and lyophilized lipoprotein fractions extracted from ostrich egg yolk (TCF-LPFo) on selected biological properties of cryopreserved sperm cells. Post-thaw percentage of motile sperm (MOT) was significantly higher (P<0.05) for TCF-HEY extender (66.3 ± 3.2%) than for TCF-LPFo extender (52.4 ± 3.4%). Moreover, there was no significant difference in the percentage of sperm with progressive motility (PMOT). Both diluents effectively preserved sperm plasma membrane integrity and mitochondrial function. However, it was observed that cryopreservation impaired the functionality of antioxidant sperm enzymes. The above was manifested by reduced SOD activity, in particular in samples preserved in the TCF-HEY extender, as well as decreased GPx activity. Both diluents inhibited the rate of lipid peroxidation in sperm plasma membrane during freezing-thawing. Our results suggest that LPFo is a satisfactory alternative to hen egg yolk in the extender used for canine sperm cryopreservation.

9

A preliminary study on antioxidant status of dog semen

80%

Kowalowka M. , Strzezek R. , Koziorowska-Gilun M. , Strzezek J.

Acta Biochimica Polonica

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2006

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tom 53

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nr Supplement 1

10

Cryopreservation of canine semen: the effect of two extender variants on the quality and antioxidant properties of spermatozoa

80%

Strzeżek R. , Koziorowska-Gilun M. , Stawiszyńska M.

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11

Mechanizmy obrony antyoksydacyjnej i oddziaływania reaktywnych form tlenu na plemniki samców zwierząt domowych

71%

Gotowiecka M. , Nizanski W. , Partyka A. , Strzezek R. , Koziorowska-Gilun M.

Medycyna Weterynaryjna

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2015

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tom 71

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nr 07

EN

The effect of reactive oxygen species (ROS) on spermatozoa can be twofold. Depending on their concentration, moment of activity, and exposure time, ROS can be an essential element in modulating many physiological processes or a cause of serious damage to the gametes. ROS of the greatest importance to the quality of semen are superoxide anion O₂⁻˙ and hydrogen peroxide – H₂O₂. Trace amounts of these compounds are essential for such processes as sperm maturation, capacitation, hyperactivation, acrosome reaction, and the fusion of the sperm with the oocyte. Their excess results in the development of oxidative stress, which significantly reduces the quality of sperm through the peroxidation of lipids, proteins, and other elements of cellular structure. The antioxidative system of semen limits the negative effects of oxidative stress on sperm. The proper function of the reproductive system and successful fertilization depend on maintaining a delicate balance between the amount of ROS and the activity of the antioxidant system.

12

Ocena wpływu stresu oksydacyjnego na nasienie zwierząt

71%

Gotowiecka M. , Nizanski W. , Partyka A. , Strzezek R. , Koziorowska-Gilun M.

Medycyna Weterynaryjna

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2015

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tom 71

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nr 12

EN

Oxidative stress has a significant impact on the quality of the mammalian semen. An excessive amount of reactive oxygen species (ROS) present in the environment of the spermatozoa, which rises as a result of prooxidative-antioxidative balance disorders, leads inter alia to a decrease of semen motility, changes in the conformation of the spermatozoa membrane, acrosome damage, decrease of mitochondrial activity, and appearance of the lipid peroxidation, which can become a cause of reduced fertility or infertility. For this reason the estimation of the influence of oxidative stress on semen can turn out to be a very useful tool in diagnostics of problems with fertility. Nowadays, in order to evaluate the intensity of the oxidative stress it is possible to use the direct method and indirect method that evaluate both the amount of ROS in the semen and the increase of changes in the spermatozoa caused by this compound. A precise analysis of the changes, resulting from prooxidative-antioxidative balance disorders with use of a suitable method of analysis is a base for accurate diagnosis.

13

Changes in the expression of selected antioxidative proteins in roe deer (Capreolus capreolus) epididymis in different periods of the rutting season

71%

Lecewicz M. , Majewska A. , Wysocki P. , Koziorowska-Gilun M. , Kordan W.

Annals of Animal Science

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2017

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tom 17

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nr 2

14

Boar sperm quality in relation to presence of sp32-like protein in spermatozoa - preliminary studies

61%

Orzolek A. , Wysocki P. , Strzezek J. , Koziorowska-Gilun M. , Dziekonska A. , Kordan W.

Bulletin of the Veterinary Institute in Puławy

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2015

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tom 59

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nr 2

EN

The aim of the study was to analyse sperm proteomes of ejaculates from Polish Large White (PLW) and Polish Landrace (PL) boars and to identify differences which putatively influence semen quality. Spermatozoa protein profiles were analysed by electrophoretic methods followed by selected techniques to evaluate semen quality on the following factors: sperm motility , lipid peroxidation levels (MDA production), ATP content, activities of superoxide dismutase (SOD) and catalase (CAT), total antioxidant status (TAS), and total oxidant status (TOS) of seminal plasma. A protein with an estimated molecular weight of 30 kDa was found in spermatozoa of selected ejaculates. Mass spectrometry demonstrated that this polypeptide is most similar to proacrosin binding protein (sp32). The presence of the protein was more frequently observed in sperm extracts obtained in spring-summer period. Ejaculates containing sp32-like protein demonstrated significantly higher spermatozoa motility, lower inhibition of MDA production by seminal plasma, and higher SOD activity in seminal plasma. Boar semen which included sp32-like protein also demonstrated lower ATP levels in spermatozoa as well as higher TAS and lower TOS of seminal plasma, though the differences were not statistically significant. Ejaculates from PLW boars, with sp32-like protein present in sperm, were characterised by significantly higher sperm motility , lower ATP content in spermatozoa, and higher TAS of seminal plasma. The diminished parameters of semen quality were observed in ejaculates from PL boars that also contained the discussed protein, but the differences were not statistically significant. These findings suggest that the presence of sp32-like protein in boar spermatozoa could influence semen quality.

15

Effect of boar ejaculate fraction, extender type and time of storage on quality of spermatozoa

61%

Dziekonska A. , Swiader K. , Koziorowska-Gilun M. , Mietelska K. , Zasiadczyk L. , Kordan W.

Polish Journal of Veterinary Sciences

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2017

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tom 20

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nr 1

EN

The aim of this study was to investigate the effect the sperm-rich fraction (F1) and the post-F1 fraction (F2) on the quality of boar spermatozoa stored in a liquid state. Ejaculates were collected from three Polish Landrace boars. Each ejaculate fraction was diluted with BTS short-term extender and Safe-Cell Plus (SCP) long-term extender and stored for seven days (D1-D7) at 17°C. Analyses included sperm motility parameters, normal apical ridge (NAR) acrosomes and plasma membrane integrity (PMI). Prior to the dilution of fractions, marked changes (p<0.05) were noted between F1 and F2 in progressive motility (PMOT), velocity average pathway (VAP) and velocity straight line (VCL). After the ejaculate was diluted, the type of fraction and type of extender significantly affected (p<0.05) PMOT, being markedly higher (p<0.05) for F1 extended in BTS. No marked changes (p<0.05) were observed between F1 and F2 extended in SCP for any of the analyzed sperm quality parameters during seven days of storage. Significantly higher (p<0.05) values of sperm quality parameters were noted in F1 compared with F2 for BTS on D7 of storage. The results of the four-way ANOVA analysis indicate that boar, fraction of ejaculate, extender type and day of storage had significant effects on the quality of boar stored spermatozoa. The F1 was characterised by higher quality of spermatozoa during storage in comparison with F2 in the short-term extender. Using the long-term extender containing the proteins allowed for a better application of F2, which could be important for the pig industry.

16

Seasonal-dependent variations in metabolic status of spermatozoa and antioxidant enzyme activity in the reproductive tract fluids of wild boar/domestic pig hybrids

61%

Strzeżek J. , Kordan W. , Dziekońska A. , Fraser L. , Koziorowska-Gilun M. , Koziorowski M.

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17

Effect of boar ejaculate fraction, extender type and time of storage on quality of spermatozoa

61%

Zasiadczyk Ł. , Kordan W. , Dziekońska A. , Świąder K. , Koziorowska-Gilun M. , Mietelska K.

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18

Seasonal-dependent variations in metabolic status of spermatozoa and antioxidant enzyme activity in the reproductive tract fluids of wild boar/domestic pig hybrids

61%

Dziekonska A. , Fraser L. , Koziorowska-Gilun M. , Strzezek J. , Koziorowski M. , Kordan W.

Polish Journal of Veterinary Sciences

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2014

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tom 17

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nr 2

EN

This study investigated seasonal changes in the metabolic performance of spermatozoa and activity of the antioxidant enzymes in the seminal plasma of three wild boar/domestic pigs (aged 1.5 to 2.5 years) and the activity of the antioxidant enzymes in fluids of the cauda epididymidis and vesicular glands from 16 wild boar/domestic pig hybrids (aged 1 to 3 years). Parameters of the sperm metabolic activity, such as total motility, mitochondrial functions, and measurements of oxygen uptake, ATP content and L-lactate production, were analyzed during the spring-summer and autumn-winter periods. Besides these sperm metabolic parameters, the sperm membrane integrity was also assessed. Total protein content and activity of the antioxidant enzymes, such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx), were measured in the reproductive tract fluids. There were no marked significant differences (P > 0.05) between the seasonal periods in terms of sperm motility, mitochondrial function and oxygen uptake; however, spermatozoa collected during the autumn-winter period exhibited higher (P < 0.05) ATP content and L-lactate production than those harvested during the spring-summer period. It was found that the vesicular gland fluid exhibited a higher level of SOD activity during the spring-summer period compared with the autumn-winter period. Furthermore, CAT activity in the seminal plasma and vesicular gland fluid was greater during the autumn-winter. Total protein content was significantly higher in the vesicular gland fluid, whereas the cauda epididymidal fluid exhibited greater SOD and GPx activities, irrespective of the seasonal period. The findings of this study confirmed seasonal-related differences in the metabolic performance of spermatozoa and activity of antioxidant enzymes of the reproductive tract of the boar/domestic pig hybrids.

19

Effects of storage in different semen extenders on the pre - freezing and post - thawing quality of boar spermatozoa

51%

Dziekonska A. , Zasiadczyk L. , Lecewicz M. , Strzezek R. , Koziorowska-Gilun M. , Fraser R. , Mogielnicka-Brzozowska M. , Kordan W.

Polish Journal of Veterinary Sciences

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2015

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tom 18

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nr 4

EN

The aim of this study was to investigate the effects of storage of semen in different commercial extenders on the pre-freezing and post-thawing quality of boar spermatozoa. Semen was diluted in BTS, Androhep (AH) and Gedil (GD), stored for 24 h at 17°C, and then frozen in accordance with the cryopreservation protocol. Analyses of the quality of spermatozoa included: motility, normal apical ridge (NAR) acrosome, plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), measurements of ATP content and activity of superoxidase dismutase (SOD) and glutathione peroxidase (GPx). Prior to the freezing process, no significant effect of the extender on the sperm quality parameters was noted. After thawing the spermatozoa it was demonstrated that the type of extender used influenced PMI, MMP, ATP content and activity of GPx. In the AH extender the percentage of spermatozoa with PMI and ATP content in spermatozoa was significantly higher (P<0.05) as compared to the BTS or GD extenders. In addition, semen stored in the AH was characterised by a statistically higher (P<0.05) percentage of spermatozoa with MMP and increased activity of GPx as compared with the BTS. The results obtained indicate that for the cryopreservation process, boar spermatozoa stored for 24 hours in liquid state can be used. However, the type of extender used prior to freezing may have a significant effect on the post- thawing quality of the spermatozoa. The AH extender better secured the quality of thawed boar spermatozoa as compared with the BTS or GD.

20

Semen quality assessments and their significance in reproductive technology

51%

Kordan W. , Fraser L. , Wysocki P. , Strzezek R. , Lecewicz M. , Mogielnicka-Brzozowska M. , Dziekonska A. , Soliwoda D. , Koziorowska-Gilun M.

Polish Journal of Veterinary Sciences

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2013

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tom 16

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nr 4

EN

Semen quality assessment methods are very important in predicting the fertilizing ability of persevered spermatozoa and to improve animal reproductive technology. This review discusses some of the current laboratory methods used for semen quality assessments, with references to their relevance in the evaluation of male fertility and semen preservation technologies. Semen quality assessment methods include sperm motility evaluations, analyzed with the computer-assisted semen analysis (CASA) system, and plasma membrane integrity evaluations using fluorescent stains, such as Hoechst 33258 (H33258), SYBR-14, propidium iodide (PI), ethidium homodimer (EthD) and 6-carboxyfluorescein diacetate (CFDA), and biochemical tests, such as the measurement of malondialdehyde (MDA) level. This review addresses the significance of specific fluorochromes and ATP measurements for the evaluation of the sperm mitochondrial status. Laboratory methods used for the evaluation of chromatin status, DNA integrity, and apoptotic changes in spermatozoa have been discussed. Special emphasis has been focused on the application of proteomic techniques, such as two-dimensional (2-D) gel electrophoresis and liquid chromatography mass spectrometry (LC-MS/MS), for the identification of the properties and functions of seminal plasma proteins in order to define their role in the fertilization-related processes.