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Freezability and fertility of frozen-thawed boar semen supplemented with ostrich egg yolk lipoproteins

L. Fraser Ł. Zasiadczyk J. Strzeżek R. Strzeżek K. Karpiesiuk
Polish Journal of Veterinary Sciences | 2018 | vol. 21 | No 2 | 255–263 | DOI: 10.24425/119046
Keywords boar egg yolk lipoproteins cryopreservation artificial insemination
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Abstract

Lipoproteins, isolated from ostrich egg yolk (LPFo), provide excellent protection for boar spermatozoa against cryo-induced damage. The present study was performed to investigate the effects of LPFo on the freezability and fertilizing capacity of frozen-thawed (FT) boar semen after post-cervical artificial inseminations (post-CAIs). Semen, collected from 7 Polish Large White (PLW) and 4 Polish Landrace (PLR), was frozen in an extender containing LPFo. Post-CAIs were performed in 38 multiparous sows, using a catheter-cannula kit. Sows were inseminated 2× within one oestrus, and fertility parameters were recorded after farrowing. Neither boar (within breed) nor breed affected the quality of the pre-freeze (PF) semen, such as total motility (TMOT), mitochondria membrane potential (MMP), plasma membrane integrity (PMI), osmotic resistance test (ORT) and DNA fragmentation. Differences in the freezability of boar semen were observed among the boars, whereas there were no marked breed effects. Post-thaw TMOT markedly declined over storage time in most of the boars, particularly at 60 min after thawing. Inseminations of post-weaned oestrus sows resulted in pregnancy and farrowing rates of 84.2% and 81.6%, respectively. Neither the mean number of piglets born (NB) nor the mean number of piglets born alive (NBA) was affected by boar or breed. The total number of piglets born was 365, resulting in 11.8 NB piglets, whereas the total number of piglets born alive was 353, with 11.4 NBA piglets per litter. The findings of this study reaffirm the variations in the freezability of boar semen. In this study the supplementation of ostrich egg yolk lipoproteins to the freezing extender of boar semen produced high proportions of functionally viable FT spermatozoa that were capable of providing acceptable fertility results after post-CAIs in multiparous sows.
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Authors and Affiliations

L. Fraser
Ł. Zasiadczyk
J. Strzeżek
R. Strzeżek
K. Karpiesiuk

Effect of post-thaw supplementation of fractionated seminal plasma on survival of boar spermatozoa

K. Wasilewska-Sakowska Ł. Zasiadczyk L. Fraser J. Strzeżek K. Karpiesiuk
Polish Journal of Veterinary Sciences | 2019 | vol.22 | No 3 | 617–625 | DOI: 10.24425/pjvs.2019.129972
Keywords boar semen fractionated seminal plasma cryopreservation
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Abstract

In our recent study we demonstrated that the holding of fresh semen in fractionated seminal plasma (SP1, >40 kDa; SP2, <40 kDa), obtained by gel filtration chromatography, significantly improved the sperm quality characteristics following cryopreservation (Wasilewska-Sakowska et al. 2019). In this study we investigated the effect of post-thaw (PT) supplementation of fractionated SP (SP1 and SP2) on the survival of spermatozoa from boars with good and poor semen freezability, GSF and PSF, respectively. Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) analysis showed distinct differences in the protein profiles of SP1 and SP2 from boars with GSF or PSF regarding the number of protein spots. Sperm motility characteristics and the motion patterns, assessed using the computer-assisted sperm analysis (CASA) system, were markedly higher in PT semen supplemented with SP1 and SP2 from boars with GSF. Post-thaw supplementation of either SP1 or SP2 from boars with GSF significantly improved mitochondrial function, plasma membrane and acrosome integrity, and viability during storage. The findings of this study have confirmed that the presence of protective protein components in varying abundance in either fractionated SP from boars with good freezability ejaculates significantly improved the sperm survival following PT storage.

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Authors and Affiliations

K. Wasilewska-Sakowska
Ł. Zasiadczyk
L. Fraser
J. Strzeżek
K. Karpiesiuk

The effect of luteolin on spermatological parameters, apoptosis, oxidative stress rate in freezing rabbit semen

S.A. Akarsu T.C. Acısu İ.H. Güngör A. Çakır Cihangiroğlu R.H. Koca G. Türk M. Sönmez S. Gür F. Fırat H.E. Esmer Duruel
Polish Journal of Veterinary Sciences | 2023 | vol. 26 | No 1 | 91-98 | DOI: 10.24425/pjvs.2023.145010
Keywords rabbit semen cryopreservation luteolin oxidative stress apoptosis
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Abstract

The aim of the present study was to determine the effects of Luteolin (LUT) on semen quality, oxidative stress, apoptosis, acrosomal integrity, mitochondrial membrane potential and dead sperm ratio in rabbits. Ejaculates from six New Zealand rabbits were collected, evaluated and pooled. The pooling was divided into five groups as control (no additive) LUT 25 μM, LUT 50 μM, LUT 100 μM and LUT 200 μM and LUT added. It was then filled into a falcon tube with Tris-based extender at a final concentration of approximately 35 x 106 spermatozoa. Diluated rabbit semen samples were drawn into frozen and thawed. Frozen semen straws were thawed at 37°C in 30 seconds. According to our findings, no statistical difference was found between all doses of luteolin and the control group in the CASA (computer assisted sperm analysis) analysis performed at 4°C. However, total motility, progressive motility and rapid sperm percentage were found to be higher in the frozen and thawed rabbit semen at a dose of LUT 50 μM compared to the other groups (p<0.05). While amplitude of lateral head displacement (ALH) and beat cross-frequency (BCF) values were found at the lowest dose of LUT 200 μM, a statistically significant difference was observed between the other groups. When the flow cytometry results were examined, no statistical difference was found between the rate of dead sperm, acrosomal integrity, mitochondrial membrane potential and apoptosis rate. Morever, the H2O2 percentage was found to be lower in all experimental groups compared to the control group (p<0.001). In conclusion, the addition of LUT in long-term storage of rabbit semen provided a protective effect for spermatozoa with its antioxidative properties against damage caused by cryopreservation.
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Bibliography

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Authors and Affiliations

S.A. Akarsu
1
T.C. Acısu
2
İ.H. Güngör
2
A. Çakır Cihangiroğlu
2
R.H. Koca
3
G. Türk
2
M. Sönmez
2
S. Gür
2
F. Fırat
2
H.E. Esmer Duruel
4
  1. Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, Atatürk University, 25240 Yakutiye /Erzurum, Turkey
  2. Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, Fırat University, 23200, Elazığ, Turkey
  3. Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, Bingöl University, Selahaddin-i Eyyübi Neighbourhood, University Street No: 1, 12000, Bingöl, Turkey
  4. Elbistan Vocational School, Kahramanmaraş İstiklal University, Şehit Astsubay Ömer Halisdemir Street No:5, Doğan Neighbourhooh,46340, Elbistan, Kahramanmaraş, Turkey

The cryoprotective potential of propolis supplemented in frozen-thawed bull semen; biochemical and physiological findings

D. Yeni M.F. Gülhan M.E. İnanç F. Avdatek Ş. Güngör R. Türkmen P.B. Tuncer U. Taşdemir
Polish Journal of Veterinary Sciences | 2022 | vol. 25 | No 1 | 5-12 | DOI: 10.24425/pjvs.2022.140834
Keywords cryopreservation DNA damage oxidative stress propolis
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Abstract

In this study, the cryoprotective effect of different doses of propolis (P) on bull semen, which has solid pharmacological properties thanks to its rich phenolic components, was investigated biochemically and physiologically. Semen samples were collected from Simmental breed bulls via the artificial vagina and pooled. After dividing into five groups, control (C: no additive) and four different P (200, 100, 50, and 25 μg/mL) groups, the final concentration was diluted to 16×106 per straw. Semen samples were equilibrated at 4°C for approximately 4 hours, then placed in French straws and frozen. After thawing, sperm motility and kinetic parameters, DNA integrity by single-cell gel electrophoresis, sperm abnormalities by liquid fixation, and lipid peroxidation levels by the colorimetric method was analyzed by Computer-Assisted Semen Analyzer. P added to the diluent showed no effect on motility and kinetic parameters at P25 and P50 (p>0.05), while P100 and P200 had a negative effect (p<0.001). The addition of P (25 and 50) showed a treatment effect on tail abnormality compared to C (p<0.05). Especially P50 had a positive effect on tail length, tail DNA, and tail movement, while P100 and P200 caused DNA damage (p<0.001). MDA levels increased in all P dose groups compared to C (p<0.001). This study has clearly demonstrated that P25 and P50 supplements could be used therapeutically to treat sperm tail abnormalities and prevent DNA damage in post-thawed bull sperm.
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Authors and Affiliations

D. Yeni
1
M.F. Gülhan
2
M.E. İnanç
3
F. Avdatek
1
Ş. Güngör
3
R. Türkmen
4
P.B. Tuncer
5
U. Taşdemir
6
  1. Faculty of Veterinary Medicine, Department of Reproduction and Artificial Insemination, Afyon Kocatepe University, Ahmet Necdet Sezer Campus, 03200, Afyonkarahisar, Turkey
  2. Department of Medicinal and Aromatic Plants, Vocational School of Technical Sciences, Aksaray University, Hacilar Harmanı Street, 12. Boulevard No: 2, 68100, Aksaray, Turkey
  3. Faculty of Veterinary Medicine, Department of Reproduction and Artificial Insemination, Mehmet Akif Ersoy University, Istiklal Campus, 15030, Burdur, Turkey
  4. Afyon Kocatepe University, Faculty of Veterinary Medicine, Department of Pharmacology and Toxicology, Ahmet Necdet Sezer Campus, 03200, Afyonkarahisar, Turkey
  5. Technical Sciences Vocational School, Mersin University, Çiftlikköy Campus, 33343, Yenişehir, Mersin, Turkey
  6. Faculty of Veterinary Medicine, Department of Reproduction and Artificial Insemination, Aksaray University, Central Campus, 68100, Aksaray, Turkey

Post-thaw quality of boar spermatozoa is affected by ejaculate fractions and extenders

Ł. Zasiadczyk K. Kurpanik L. Fraser W. Kordan
Polish Journal of Veterinary Sciences | 2024 | vol. 27 | No 1 | 147-150 | DOI: 10.24425/pjvs.2024.149345
Keywords boar cryopreservation ejaculate fractions semen extender
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Abstract

The aim of this study was to investigate the effect of different extenders on the post-thaw (PT) quality of sperm originating from the sperm-rich fraction (SRF) and post-sperm-rich fraction (PSRF) of boar ejaculate. Motility and velocity parameters, analyzed using a computer-assisted semen analysis (CASA) system, and membrane integrity parameters were markedly higher in frozen-thawed (FT) spermatozoa of the SRF in both the Belstville Thawing Solution (BTS) and Androhep Plus (AHP) extenders, irrespective of the post-thaw (PT) storage time. Furthermore, reduced cryo-survival was more marked in FT spermatozoa of the PSRF in both extenders following storage for 60 min. It was found that the SRF-stored samples in the AHP extender for 60 min exhibited significantly higher percentages of spermatozoa with total motility, mitochondrial function and acrosome integrity than those stored in the BTS extender. The findings of this study confirm that components of the ejaculate fractions and extender have varying effects on the cryo-survival of boar spermatozoa.
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Authors and Affiliations

Ł. Zasiadczyk
1
K. Kurpanik
1
L. Fraser
1
W. Kordan
1
  1. Department of Animal Biochemistry and Biotechnology, Faculty of Animal Bioengineering, University of Warmia and Mazury in Olsztyn, Oczapowskiego 5, 10-719 Olsztyn, Poland

Lipid peroxidation in avian semen

A. Partyka A. Babapour M. Mikita S. Adeniran W. Niżański
Polish Journal of Veterinary Sciences | 2023 | vol. 26 | No 3 | 497–509 | DOI: 10.24425/pjvs.2023.145050
Keywords avian semen lipid peroxidation oxidative stress semen cryopreservation
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Abstract

The main cause of sperm chromatin damage is oxidative stress related to embryo development failure and adult infertility in mammals and also avian. Oxidative stress results in lipid peroxidation (LPO) causing cell damage. Lipid peroxidation is the oxidation of polyunsaturated fatty acids (PUFAs) in biological systems and causes changes in the physical structure and characteristics of the cell membrane. Due to the high amounts of PUFAs in the avian sperm membrane, its sperm seem susceptible to pe-roxidative damage and is a substantial factor in the fertilization capacity of sperm. The most commonly used methods for measuring LPO or its by-products, such as malondialdehyde (MDA) and 4-hydroksy-2-nonenal (4-HNE), in bird semen are based on the colorimetric method TBARS (thiobarbituric acid reactive substances) and on the use of a fluorescence probe (CC 11-BODIPY 581/591) as a marker to evaluate membrane lipid peroxidation. This review aims first to introduce LPO in avian semen and its effects on avian sperm and second to summarize the commonly applied methods of evaluating LPO and its damage in fresh and stored avian semen.
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Authors and Affiliations

A. Partyka
1
A. Babapour
2
M. Mikita
1
S. Adeniran
3
W. Niżański
1
  1. Wroclaw University of Environmental and Life Sciences, Faculty of Veterinary Medicine, Department of Reproduction and Clinic of Farm Animals, pl. Grunwaldzki 49, 50-366 Wroclaw, Poland
  2. University of Tabriz, Faculty of Veterinary Medicine, Department of Basic Sciences, Tabriz, Iran
  3. Mountain Top University, College of Basic and Applied Sciences, Department of Biological Sciences, Ogun State, Nigeria

Interactions of ostrich egg yolk lipoproteins with seminal plasma of fractionated ejaculates and their effects on post-thaw boar semen quality

L. Fraser Ł. Zasiadczyk J. Strzeżek W. Kordan A. Mańkowska
Polish Journal of Veterinary Sciences | 2019 | vol.22 | No 3 | 463-473 | DOI: 10.24425/pjvs.2019.129306
Keywords boar semen ejaculate fractions egg yolk lipoproteins cryopreservation
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Abstract

Electrophoretic methods were used to identify protein complexes formed between ostrich egg yolk lipoprotein fractions (LPFo) with seminal plasma (SP) of fractionated ejaculates, and to investigate the effect of these complexes on boar semen quality after cryopreservation. Chromatographic SP fractions (F1, F2 and F3), with or without LPFo solution, were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. Comparative electrophoretic analyses of the SP revealed marked differences in the SDS-PAGE protein profiles among boars. Electrophoretic analyses showed that the interactions of LPFo with SP resulted in the appearance of high-intensity protein bands. Spermatozoa were exposed to SP chromatographic fractions originating from F1, F2 and F3, and the whole SP (wSP) before being frozen. Spermatozoa exposed to F1 and F2 exhibited significantly higher post-thaw motility compared to those treated with either F3 or wSP. In most of the boars the proportions of membrane-intact frozen-thawed spermatozoa differed among the treatments, being significantly lower in the wSP-treated samples. The incidence of frozen-thawed spermatozoa with DNA fragmentation was less prevalent in samples exposed to F3 or the wSP. The results of this study confirmed that the interactions of LPFo with fractionated SP during the cooling period contributed to alterations in the sperm membranes, rendering them less susceptible to temperature-related injury.

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Authors and Affiliations

L. Fraser
Ł. Zasiadczyk
J. Strzeżek
W. Kordan
A. Mańkowska

Quality parameters and fertility of ram semen cryopreserved in egg yolk and soybean lecithin supplemented extenders

P. Gogol M. Bryła M. Trzcińska M. Bochenek
Polish Journal of Veterinary Sciences | 2019 | vol.22 | No 1 | 177-179 | DOI: 10.24425/pjvs.2019.127084
Keywords ram semen cryopreservation soybean lecithin sperm quality fertility
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Abstract

The aim of the study was to investigate the effect of soybean lecithin as a substitute for egg yolk in milk and tris based extenders in ram semen cryopreservation. Twenty ejaculates were col- lected from four healthy, mature Wrzosówka rams (2-3 years of age). Each ejaculate was divided into four equal aliquots and diluted with four different extenders: 1) milk extender containing 5% egg yolk, 2) milk extender containing 1.5% soybean lecithin, 3) tris extender containing 20% egg yolk, 4) tris extender containing 1.5% soybean lecithin. Extended semen was loaded into 0.25 ml French straws, cooled and frozen in liquid nitrogen vapor. Total motility, curvilinear velocity, plasma membrane integrity and fertilizing ability of sperm were assessed after thawing. Total mo- tility was lower (p<0.05) in tris-soybean lecithin extender when compared to other extenders. Curvilinear velocity was higher (p<0.05) for spermatozoa cryopreserved in milk-soybean lecithin extender compared to other extenders tested. For the percentage of live sperm no significant difference was observed between extenders. The lambing rate were higher (not statistically signifi- cant) in ewes inseminated with semen doses frozen in milk-soybean lecithin extender (42.9%) than in the tris-egg yolk extender (16.7%). In conclusion, replacing the egg yolk with soybean lecithin was effective in milk but not in tris extender.

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Authors and Affiliations

P. Gogol
M. Bryła
M. Trzcińska
M. Bochenek

The quality and fertilizing ability of the ram semen cryopreserved around 40 years ago

P. Gogol
Polish Journal of Veterinary Sciences | 2022 | vol. 25 | No 4 | 631-633 | DOI: 10.24425/pjvs.2022.143544
Keywords ram semen ex situ storage gene banks cryopreservation sperm quality fertility
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Abstract

The aim of the study was to verify the quality of ram semen, frozen in 1982-1983, from the historical collection of the Bank of Biological Material of the National Research Institute of Animal Production. A total of 18 ejaculates from 3 Świniarka type rams were analyzed to assess sperm motility (subjectively), total motility, progressive motility, sperm concentration (CASA), membrane integrity (SYBR-14/PI) and chromatin structure (SCSA). In order to determine sperm fertilizing ability 49 ewes were intracervically inseminated (200×106 sperm per AI) with frozen- thawed semen 12 and 24 hours after detection of estrus. Sperm motility parameters, membrane intact spermatozoa and DFI did not differ among the analyzed rams. Spermatozoa concentration was significantly higher for ram no. 2 than for rams no. 1 and 3. The lambing rates (27.3 to 36.0%) did not differ significantly for individual rams. The ram semen, which had been stored for around 40 years, showed satisfactory quality and fertilizing capacity, allowing for its use in artificial insemination.
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Authors and Affiliations

P. Gogol
1
  1. Department of Reproductive Biotechnology and Cryoconservation, National Research Institute of Animal Production, Krakowska 1, 32-083 Balice/Kraków, Poland

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