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

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.

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

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.

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

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.

Metabolic Activity of Boar Semen Stored in Different Extenders Supplemented with Ostrich Egg Yolk Lipoproteins.

Introduction: The aim of this study was to evaluate the effect of lipoprotein fraction isolated from ostrich egg yolk (LPFo) on the metabolic activity of boar spermatozoa following liquid semen storage in different extenders and temperatures.

Material And Methods: Boar ejaculates were extended in Androhep, Beltsville thawing solution (BTS), and Martín-Rillo and Alias (MR-A) without (control) or with the addition of LPFo and stored for three days at either 5°C or 16°C. The analysed sperm parameters included total motility (TMOT), plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), oxygen consumption, and adenosine triphosphate (ATP) production.

Age and seasonal-dependent variations in the biochemical composition of boar semen.

This study investigated the effect of age- and seasonal-related variations in the composition of boar semen over a 3-year period. At the onset of 8 months of age, ejaculates were collected from four boars and allocated into three groups: 8 to 18, 19 to 30, and 31 to 42 months and were divided into two seasonal periods: autumn-winter and spring-summer. Boar variability had a significant effect on most of the analyzed semen parameters.

The activity of N-acetyl-β-hexosaminidase in boar seminal plasma is linked with semen quality and its suitability for cryopreservation.

The determination of sperm cryotolerance is an important step in the process of developing optimal techniques for the storage of boar semen. The objective of this study was to determine individual proteome variations in boar seminal plasma and spermatozoa and establish their influence on the cryotolerance of ejaculate. Sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed the presence of protein with estimated molecular weight of 90 kDa in sperm extracts from ejaculates of selected boars.

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

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).

Post-thaw sperm characteristics following long-term storage of boar semen in liquid nitrogen.

This study investigated the effect of long-term liquid nitrogen storage of semen from individual boars on post-thaw sperm characteristics. Ejaculates, collected from five Polish large white (PLW) and five Polish landrace (PLR) boars, were frozen using a standard cryopreservation protocol. Post-thaw analysis was performed within a week (Period 1) and 42-48 months (Period 2) of semen storage in liquid nitrogen.

The antigenic character of zinc-binding proteins and their localization in boar reproductive tract--a preliminary study.

In this study immunoelectrophoretic and double immunodiffusion analyses were used to investigate the antigenic character of zinc-binding proteins (ZnBPs), whereas the indirect immunofluorescence technique was used to identify their origin in boar reproductive tract. The mmunoelectrophoretic analysis of ZnBPs of the seminal plasma resulted in the appearance of three antigenic protein complexes, while specific immunoreactivity patterns of the anti-ZnBP serum were detected by double immunodiffusion analysis. Indirect immunofluorescence technique confirmed that ZnBPs were secreted by different reproductive tract tissues, suggesting their contributions to the seminal plasma.

Superoxide dismutase (SOD) in boar spermatozoa: purification, biochemical properties and changes in activity during semen storage (16°C) in different extenders.

The antioxidant system in semen is composed of enzymes, low-molecular weight antioxidants and seminal plasma proteins. Loss of enzymatic activity of superoxide dismutase (SOD) during semen preservation may cause insufficient antioxidant defense of boar spermatozoa. The aim of this study was to isolate and characterize SOD molecular forms from spermatozoa and to describe changes in SOD activity in boar sperm during preservation at 16°C.

Effect of freezing on sperm nuclear DNA.

Sperm DNA damage has a significant impact on reproductive outcomes. In recent years, the search for optimal molecular markers for the evaluation of semen quality has resulted in the increased focus on sperm nuclear DNA assessment. The primary aim of this article was to review and summarize the effects of freezing-thawing procedure on nuclear DNA integrity of boar spermatozoa.

Zinc-binding proteins from boar seminal plasma -- isolation, biochemical characteristics and influence on spermatozoa stored at 4°C.

Affinity chromatography on Chelating Sepharose Fast Flow Gel-Zn(2+) was used for fractionation of boar seminal plasma proteins. Approximately 30% of total boar seminal plasma proteins showed affinity for zinc ions (ZnBP fraction). Native electrophoresis (PAGE) of ZnBP revealed six protein fractions which separated into 27 bands under denaturing conditions (SDS/PAGE).

Boar variability affects sperm metabolism activity in liquid stored semen at 5 degrees C.

Metabolic activity of boar spermatozoa, liquid stored for three days at 5 degrees C, was measured using bioluminescence for ATP content, fluorescent assay (JC fluorochrome) of mitochondrial activity and oxygen consumption. Sperm motility and plasma membrane integrity (PMI) were simultaneously analyzed. Apart from the statistically significant effect (P < 0.

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

This study aimed to analyze seasonal variations in the antioxidant defence systems of the seminal plasma and fluids of the cauda epididymis and vesicular glands of the boar. The analyzed antioxidants included superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and total L-glutathione (GSH+GSSG). Seasonal changes in total protein content and total antioxidant status (TAS) of the seminal plasma and reproductive fluids were also analyzed.

Antioxidant defence system of boar cauda epididymidal spermatozoa and reproductive tract fluids.

Antioxidants secreted by the reproductive tract protect spermatozoa against the toxic effects of reactive oxygen species (ROS) after ejaculation. This study aimed at characterizing the level of antioxidant protection in boar cauda epididymidal spermatozoa and fluids of the cauda epididymidis, vesicular and prostate glands. Also, this study investigated the effect of a 5-h period of dialysis on the antioxidant capacity of boar seminal plasma.

Interactions of egg yolk lipoprotein fraction with boar spermatozoa assessed with a fluorescent membrane probe.

The interactions of a fluorescent membrane probe, 1-anilinonaphthalene-8-sulfonic acid (1,8-ANS), with boar spermatozoa were followed through the use of lipoprotein fraction of ostrich egg yolk (LPFo). Semen samples, extended in Kortowo 3 (K3) extender, were supplemented with 2% or 5% LPFo and stored for 3h at 16 degrees C. Additionally, cold shock-treated spermatozoa (1h at 4 degrees C) were stored in K3 extender supplemented with LPFo for 3h at 16 degrees C.

Comparison of post-thaw DNA integrity of boar spermatozoa assessed with the neutral comet assay and Sperm-Sus Halomax test kit.

In this study, we tested the hypothesis whether the neutral Comet assay (NCA) and the Sperm-Sus-Halomax (SSH) test kit could provide similar measurements of post-thaw DNA fragmentation of boar spermatozoa. Whole ejaculates or sperm-rich fractions of boar semen were frozen in an extender containing lactose, lipoprotein fractions isolated from ostrich egg yolk (LPFo), glycerol (lactose-LPFo-G) or in a standard boar semen extender (K3), without the addition of cryoprotective substances. In all boars, both the NCA and SSH test showed similar levels of post-thaw sperm DNA fragmentation in samples of the same ejaculates, regardless of the ejaculate collection procedure and extender.

Recent advances in boar semen cryopreservation.

Since 35 years ago boar semen has been frozen and used for artificial insemination (AI). However, fertility of cryopreserved porcine sperm has consistently been low as boar sperm are more sensitive to cellular stress imposed by changing osmotic balance, oxidative stress, low-temperature exposure, cryo-protectant intoxication etc. and are less able to compensate for these deficiencies at commercially applicable dosages.

High sequence homology between protein tyrosine acid phosphatase from boar seminal vesicles and human prostatic acid phosphatase.

Boar seminal vesicle protein tyrosine acid phosphatase (PTAP) and human prostatic acid phosphatase (PAP) show high affinity for protein phosphotyrosine residues. The physico-chemical and kinetic properties of the boar and human enzymes are different. The main objective of this study was to establish the nucleotide sequence of cDNA encoding boar PTAP and compare it with that of human PAP cDNA.

Characteristics of antioxidant system in dog semen.

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.

Extracellular superoxide dismutase of boar seminal plasma.

Superoxide dismutase (SOD) is an enzymatic component of the antioxidant defense system that protects spermatozoa by catalysing the dismutation of superoxide anions to hydrogen peroxide and oxygen. Age and season effects on SOD activity in the seminal plasma were measured in boars at the onset of 8 months through a 35-month period. It was found that age-related changes in SOD activity in the seminal plasma were markedly higher in boars less than 2 years of age.

Fertilizing capacity of boar semen frozen in an extender supplemented with ostrich egg yolk lipoprotein fractions--a pilot study.

A limited field trial was performed to evaluate the fertilizing capacity of boar spermatozoa frozen in an extender supplemented with lipoprotein fractions isolated from ostrich egg yolk (LPFo). Boar semen, diluted in an extender containing lactose with lyophilized lipoprotein fractions, glycerol and Orvus Es Paste (lactose-LPFo-G), was frozen using a controlled programmable freezer. Sperm characteristics, such as motility, plasma membrane and acrosome integrity, and mitochondrial function were monitored.

Is there a relationship between the chromatin status and DNA fragmentation of boar spermatozoa following freezing-thawing?

In this study a radioisotope method, which is based on the quantitative measurements of tritiated-labeled actinomycin D ((3)H-AMD) incorporation into the sperm nuclei ((3)H-AMD incorporation assay), was used to assess the chromatin status of frozen-thawed boar spermatozoa. This study also tested the hypothesis that frozen-thawed spermatozoa with altered chromatin were susceptible to DNA fragmentation measured with the neutral comet assay (NCA). Boar semen was diluted in lactose-hen egg yolk-glycerol extender (L-HEY) or lactose ostrich egg yolk lipoprotein fractions-glycerol extender (L-LPFo), packaged into aluminum tubes or plastic straws and frozen in a controlled programmable freezer.

Dialysis of boar semen prior to freezing-thawing: its effects on post-thaw sperm characteristics.

Low-molecular weight components of the seminal plasma have a detrimental effect on sperm function. The present study was undertaken to evaluate the effect of the removal of low-molecular weight components by dialysis on sperm characteristics prior to and after freezing. Semen, collected from 5 boars, was extended in Kortowo-3 extender (K-3, Poland) and cooled for 3h (control non-dialysis) or dialyzed for 5h in semi-permeable dialysis bags of 12-14kDa molecular weight cut-off prior to freezing.

Application of biochemical markers for identification of biological properties of animal semen.

The use of biochemical markers for identification of biological properties of semen will help to develop new criteria that are accurate and objective in predicting and improving male fertility. Understanding and controlling the mechanisms involved in fertility is a key challenge, which is of fundamental importance in successful animal reproductive performance. Moreover, unraveling the unique molecular mechanism associated with sperm function might have considerable diagnostic value in the evaluation of male infertility.