73 SUPPLEMENTATION OF FROZEN BOAR SEMEN WITH β-MERCAPTOETHANOL INCREASES THE INCIDENCE OF INTACT CELLS AFTER THAWING

2008 ◽  
Vol 20 (1) ◽  
pp. 117
Author(s):  
H. Funahashi ◽  
S. Yamaguchi ◽  
W. Fujii ◽  
T. Murakami
Keyword(s):  
Liquid Nitrogen ◽  
Dna Fragmentation ◽  
Egg Yolk ◽  
Molecular Probes ◽  
Sperm Cells ◽  
Freezing And Thawing ◽  
Intact Cells ◽  
Boar Semen ◽  
Post Hoc ◽  
Boar Spermatozoa

During the process of freezing and thawing of boar spermatozoa, a large number of the cells appear to be injured by some stresses such as osmotic forces and oxidation, causing reduced viability and penetrability. β-Mercaptoethanol (bME), a strong reducing agent, may ease oxidative stress and rescue sperm cells from those injuries. The aim of this study was to determine the effect of the presence of bME during freezing and thawing of boar spermatozoa on the viability and acrosome status of the sperm cells. Semen samples were collected from 3 boars; only samples with a high motility (more than 80%) were used for this experiment. Each sample was diluted 1:1 with modified Modena solution and kept overnight at 15�C. After centrifugation at 800g for 10 min, the diluent supernatant was removed; spermatozoa were re-suspended at 2 � 109 cells mL–1 in the first diluent (8.8% trehalose solution containing 20% egg yolk and antibiotic) supplemented with 0, 25, or 50 µm bME, and then cooled to 5�C over 2–3 h. At 5�C, semen samples were further diluted 1:1 with the second diluent (same as the first diluent + 5% glycerin + 1.48% Orvus ES Paste (Equex STM; Minitube, Verona, WI, USA)) supplemented with 0, 25, and 50 µm bME, respectively. After packaging the semen into 0.5-mL straws, it was frozen by keeping the straws 4 cm above the surface of liquid nitrogen for 15 min and then storing them in liquid nitrogen until use. After thawing at 37�C for 30 s, semen samples were re-suspended in 10 mL of BTS solution containing 1.15 mm caffeine and 4 mm Ca chloride, and incubated at 37�C under 5% CO2 in air for 90 min. Viability, DNA fragmentation, and acrosome status of spermatozoa were assessed by flow cytometry after staining with SYBR�14/PI (Molecular Probes, Inc., Eugene, OR, USA), acridine orange, and PNA/PI, respectively. Statistical analyses of data from at least 3 replicated trials were carried out by ANOVA and Fisher's protected least-squares difference (PLSD) post-hoc test. Just after thawing, no differences in viability (45.6–51.1%; P = 0.67), DNA fragmentation (0.7–0.9%; P = 0.76), and acrosome status (intact acrosome: 79.2–83.0%; P = 0.26) of the spermatozoa were observed when sperm cells were frozen and thawed in 0, 25, and 50 µm bME. After culture for 90 min, however, the incidence of spermatozoa with an intact acrosome was significantly higher (P < 0.05) when the semen was frozen and thawed in the presence of 50 µm bME (70.9%), compared with 0 (61.7%) and 25 µm bME (61.0%). Chlortetracycline (CTC) analyses were peformed to confirm that the incidence of intact spermatozoa was higher (P < 0.01) in 50 µm bME (67.6%) than that of non-supplementation controls (51.4%). These results demonstrate that supplementation of semen with 50 µm bME during freezing and thawing processes reduces acrosome damage of boar spermatozoa.

scholarly journals Cryopreservation of boar semen

2020 ◽  
Vol 65 (No. 4) ◽  
pp. 115-123
Author(s):  
Marija Jovičić ◽  
Eva Chmelíková ◽  
Markéta Sedmíková
Keyword(s):  
Animal Species ◽  
Semen Quality ◽  
Egg Yolk ◽  
High Rate ◽  
Freezing And Thawing ◽  
Long Term Storage ◽  
Boar Semen ◽  
Boar Spermatozoa ◽  
Term Storage

Sperm cryopreservation is the best technology for long-term storage of the semen. However, the damage of boar spermatozoa by cryopreservation is more severe than in other animal species and a standardized freezing protocol for efficient cryopreservation has not been established yet. Semen quality and freezability vary greatly between breeds as well as between individual boars and even the season. Boar spermatozoa are sensitive to low temperatures; they sustain damage and a high rate of mortality and freezing/thawing the boar semen may strongly impair the sperm function and decrease the semen quality. The freezability of boar semen can be influenced by a cryopreservation procedure, and also by using various additives to freezing and thawing extenders such as antioxidants. In order to obtain acceptable results after thawing the boar semen, it is necessary to combine an optimal amount of additives (glycerol, egg yolk, sugars, antioxidants), cooling and warming velocities.

The use of butylated hydroxytoluene in cryopreservation of boar semen

2016 ◽  
Vol 12 (2) ◽  
pp. 21-28
Author(s):  
Monika Trzcińska ◽  
Magdalena Baryła
Keyword(s):  
Dna Fragmentation ◽  
Sperm Quality ◽  
Annexin V ◽  
Egg Yolk ◽  
Butylated Hydroxytoluene ◽  
Preimplantation Embryos ◽  
Progressive Motility ◽  
Boar Semen ◽  
Boar Spermatozoa

The objective of the study was to determine the effect of butylated hydroxytoluene (BHT) on the quality and fertilizing capacity of frozen-thawed (FT) boar semen. Semen from five boars (36 ejaculates) was resuspended in lactose-egg yolk-glycerol extender supplemented with 0 (control), 1.0 (R1), 1.5 (R2) or 2.0 mM BHT (R3). Sperm quality was assessed based on motility (CASA; TM: total motility; PM: progressive motility), phosphatidylserine (PS) translocation across the plasma membrane (Annexin-V-FLuos Staining Kit) and DNA fragmentation (TUNEL Assay). The FT semen was also used for intrauterine artificial insemination (AI) of synchronized gilts. The fertilizing capacity of the FT semen was assessed on the basis of the gilt insemination rate and the number of morphologically normal embryos. The quality of the preimplantation embryos was determined by observing a TUNEL-positive reaction. The highest percentage of progressive motile and viable spermatozoa was noted in extender R3 (74.8 ±4.4% and 63.7 ±5.8%), as compared with the control (38.3 ±2.8% and 36.1 ±2.6%). The addition of BHT to the extender did not increase early apoptotic changes in the frozen-thawed spermatozoa with respect to the control. Irrespective of the variant of the extender, cryopreservation and thawing did not induce fragmentation in the boar spermatozoa. The highest number of morphologically normal embryos from inseminated gilts was observed in the case of semen cryopreserved in extender supplemented with 1.5 mM BHT. No significant differences were observed in DNA fragmentation in the expanded blastocysts from gilts inseminated with FT semen cryopreserved in the extenders analysed.

75 GLUCOSE CONCENTRATION OF FREEZING EXTENDER MODULATES THE TYROSINE PHOSPHORYLATION PATTERN OF FROZEN-THAWED BOAR SPERMATOZOA

2009 ◽  
Vol 21 (1) ◽  
pp. 138
Author(s):  
J. E. Rodríguez-Gil ◽  
M. Hernández ◽  
M. M. Rivera ◽  
L. Ramió-Lluch ◽  
J. Ballester ◽  
...  
Keyword(s):  
Protein Phosphorylation ◽  
Tyrosine Phosphorylation ◽  
Glucose Concentration ◽  
Egg Yolk ◽  
Freezing And Thawing ◽  
Precise Control ◽  
Boar Sperm ◽  
Phosphorylation Pattern ◽  
Thawing Process ◽  
Boar Spermatozoa

The optimization of freezing extenders is an essential issue for enhancing boar sperm cryosurvival. The aim of the present study was to disclose the role of glucose concentration of freezing extender on the metabolic activity of frozen–thawed spermatozoa. To achieve it, pooled sperm-rich ejaculate fractions from 5 mature and fertile boars (3 ejaculates per boar) were collected using the gloved-hand method. After centrifugation (2400g for 3 min), the sperm pellet was split into 7 aliquots. The aliquots were diluted to a final concentration of 1 × 109 sperm mL–1, in a Tris-citric extender supplemented with 20% egg-yolk, 3% glycerol, and 0, 0.05, 2, 4, 10, 55, or 185 mm glucose. All the extenders were adjusted to a pH of 6.8 and 310 mOsm kg–1 to avoid osmolarity effects. Extended semen samples were dispensed into 0.5-mL straws, and frozen in a programmable cell freezer at 20°C min–1. Thawing was carried out in a water bath at 37°C for 20 s. Afterward, an analysis of protein phosphorylation in tyrosine residues was carried out through bi-dimensional electrophoresis followed by a Western blot analysis. This analysis indicated that sperm samples frozen in extenders without glucose showed specific changes in the tyrosine phosphorylation pattern compared with fresh sperm. Furthermore, the addition of glucose in increasing concentrations to the freezing extender was accompanied by a concentration-dependent decrease in the overall tyrosine phosphorylation pattern, especially in proteins with a molecular weight ranging from 150 to 200 kDa and an acidic isoelectric point (pI). The maximal decrease was observed in spermatozoa frozen in the extender containing 185 mm glucose, in which an additional decrease in the tyrosine phosphorylation of proteins ranging from 60 to 80 kDa, and a basic pI was also observed. These results suggest that glucose is a modulator in the resistance of boar sperm to support freezing and thawing process, because the precise protein phosphorylation pattern of spermatozoa is directly linked to their functional status. In this way, a precise control of the glucose concentration of the freezing extender would be required to improve boar sperm cryoresistance. Supported by CICYT (AGL2005-00760 and AGL2004-04756-C02-02/GAN), Madrid and GERM (04543/07), Murcia, Spain.

INFLUENCE OF GLYCEROL CONCENTRATION AND FREEZING TO −79 C ON OXYGEN UPTAKE AND LIVABILITY OF BOAR AND BULL SPERMATOZOA

1972 ◽  
Vol 52 (1) ◽  
pp. 65-72 ◽  
Author(s):  
L. M. SANFORD ◽  
G. J. KING ◽  
J. W. MACPHERSON
Keyword(s):  
Oxygen Uptake ◽  
Incubation Period ◽  
Skim Milk ◽  
Egg Yolk ◽  
Freezing Process ◽  
Glycerol Concentration ◽  
Motile Cells ◽  
Boar Semen ◽  
Boar Spermatozoa

Boar and bull spermatozoa were diluted in a skim milk–egg yolk–glucose extender containing 0, 7.5, or 15% glycerol (v/v) and incubated aerobically for 6 hr at 37 C. Other partially diluted boar semen samples were cooled to 5 C. Glycerol was added to a final concentration of 0, 7.5, and 15%. Samples were frozen to −79 C, rewarmed, and incubated for 3 hr at 37 C. The presence of glycerol in the extender depressed (P < 0.01) the oxygen uptake by nonfrozen boar and bull spermatozoa during the 6-hr incubation period. The reduction of oxygen uptake by semen samples increased as the level of glycerol in the extender increased. There was a corresponding decrease (P < 0.01) in the number of motile cells at the conclusion of the incubation period. Glycerol appeared to have more of a detrimental effect on boar spermatozoan oxygen uptake. The rate of oxygen uptake by boar semen samples postfreezing was extremely depressed, suggesting that spermatozoa surviving the freezing process metabolize at a much lower rate than normal. Active progressive motility of most of the surviving boar spermatozoa ceased within 1–2 hr of incubation under the in vitro conditions of this experiment.

scholarly journals Determination of the optimal concentration of Minitube Equex paste for boar semen cryopreservation based on sperm motility characteristics

2020 ◽  
Vol 52 (3) ◽  
Author(s):  
Junpen Suwimonteerabutr ◽  
Morakot Nuntapaitoon ◽  
Padet Tummaruk
Keyword(s):  
Sperm Motility ◽  
Egg Yolk ◽  
Group Iv ◽  
Optimal Concentration ◽  
Freezing And Thawing ◽  
Semen Cryopreservation ◽  
Group Iii ◽  
Group I ◽  
Boar Semen ◽  
Group Ii

Equex paste is a non-permeating cryoprotective agent (CPA) that improved post-thaw survival of spermatozoa during boar semen cryopreservation. However, Equex paste produced by Nova Chemical Sales Inc. (MA, USA) is not currently available. The aim of the present study was to determine the optimal concentration of Minitube Equex paste (Minitube, Tiefenbach, Germany) for boar semen cryopreservation in comparison with Nova Equex STM paste (control). Fifteen ejaculates from 12 mature boars were collected by the glove-hand method. Each ejaculate was aliquoted and cryopreserved in base freezing extender III as Tris-citrate egg yolk (TEY) extender plus 9.0% glycerol classified into four groups. Group I was the control and included only 1.5% Nova Equex STM paste. Groups II, III and IV were the experiment groups, and contained different concentrations of Minitube Equex paste (Group II: 1.5%; Group III: 1.7%; and Group IV: 1.9%) added to the freezing extender III. After freezing and thawing, sperm motility characteristics were evaluated by Sperm Class Analyzer® incubated at 37 °C for 0 (10 min), 1 and 2 h post-thawing. In Group IV after thawing at 0 h, rapid velocity and the velocity curved line were significantly higher than in Groups II and III (P &lt; 0.05) but did not differ from Group I. Moreover, after thawing at 1 h, LIN (linearity) in Group IV was higher than in Group II (P &lt; 0.05), but did not differ from the other groups. In conclusion, the most suitable concentration of Minitube Equex paste in the current protocol was 1.9% supplemented with 9.0% glycerol in TEY-based freezing extender III, based on the conformity between data from manual guides and the observed sperm motility characteristics results.

142 RELATIONSHIP BETWEEN APOPTOSIS IN BOAR SEMEN AND DNA FRAGMENTATION IN PORCINE EMBRYOS

2007 ◽  
Vol 19 (1) ◽  
pp. 188
Author(s):  
M. Bryla ◽  
M. Trzcinska ◽  
Z. Smorag
Keyword(s):  
Dna Fragmentation ◽  
Caspase 3 ◽  
Molecular Probes ◽  
Developmental Competence ◽  
Preimplantation Embryos ◽  
Dna Integrity ◽  
Research Committee ◽  
Boar Semen ◽  
Live Sperm

Although apoptosis in somatic cells and in spermatocytes and spermatids in vivo is well established, the presence and significance of apoptosis in ejaculated animal sperm and its correlation with developmental competence of preimplantation embryos is still unresolved. The aim of this experiment was to study the relationship between apoptosis in boar semen and DNA fragmentation in porcine embryos. Two ejaculates from the same boar were used in the experiment. Both fresh ejaculates were analyzed using Vybrant Apoptosis Assay Kit #4 (Molecular Probes, Inc., Eugene, OR, USA). Then one of them was diluted in Biosolwens plus extender, stored for 5 days at 15°C and analyzed using YO-PRO-1/PI assay, which detect changes in the membrane of boar spermatozoa, based on the slight increase of membrane permeability. Both fresh and stored semen were used for insemination, of superovulated gilts (8 per group). After 5.5 days of insemination embryos were flushed out of the uterus and DNA integrity of obtained embryos were analyzed. DNA fragmentation and caspase-3 activity were detected in embryos using kits, Roche Diagnostics (Mannheim, Germany) and PhiPhiLux G2D2 (Calbiochem, San Diego, CA, USA), respectively. For both the fresh and stored semen, 3 groups of sperm were observed under a fluorescence microscope. In the fresh semen, 3 and 2% of apoptotic sperm, 13 and 9% of necrotic sperm, 84 and 89% of live sperm in first and second ejaculated, respectively, were observed. In stored semen, 14% of apoptotic sperm, 27% of necrotic, and 59% of live sperm were noted. In total, 141 expanded blastocysts for DNA fragmentation were analyzed. The results are summarized in Table 1. In conclusion, apoptosis in fresh boar semen was lower than in stored semen and was correlated with the TUNEL nucleus index in blastocysts. The expression of caspase-3 was positively correlated with cells positive for TUNEL. Table 1. Relationship between apoptosis in boar semen and DNA fragmentation in porcine embryos This study was supported by Polish Research Committee, grant no. 2 P06D 024 30.

10 Evaluation of Frozen Sperm Quality After Single Layer Centrifugation with Percoll Plus® of Fresh Bull Semen

2018 ◽  
Vol 30 (1) ◽  
pp. 144
Author(s):  
F. N. Marqui ◽  
A. Martins ◽  
T. E. Cruz ◽  
T. I. U. Berton ◽  
C. P. Freitas-Dell'Aqua ◽  
...  
Keyword(s):  
Artificial Insemination ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Single Layer ◽  
Annexin V ◽  
Egg Yolk ◽  
Molecular Probes ◽  
Becton Dickinson ◽  
Time Curve ◽  
Intact Cells

The single layer centrifugation (SLC) with Percoll Plus® (PP; GE Healthcare, Uppsala, Sweden) before freezing is not a common technique used for selection of spermatozoa in bovine. Thus, this study aimed to verify the effect of SLC with PP before freezing on integrity of plasma and acrosome membranes (IPAM), phospholipid translocation (PT), and mitochondrial membrane potential (MMP) of frozen–thawed bull sperm. Three Nellore bulls housed at the Tairana Artificial Insemination Station were used. The ejaculates (6/bull) were collected by artificial vagina and assessed for sperm motility, concentration, and morphology. Then, the sperm were pooled and ~1 billion spermatozoa, either diluted [D; 1:2 (v/v)] in freezing extender (FE; tris, fructose, citric acid, egg yolk and antibiotics, without glycerol) or undiluted (UN), were placed on top of a 9-mL column of PP (in 15-mL centrifuge tubes) at concentrations of 70% or 90%, to form the 70D, 70UD, 90D, and 90UD treatment groups. After centrifugation at 839 × g for 13 min, except for the control (C), the supernatant was discarded and the pellet diluted in FE (plus glycerol) to a final concentration of 50 × 106 spermatozoa mL−1. Afterward, 0.5-mL straws were filled, cooled for 5 h at 4°C, and frozen in a programmable freezer (Digitcool, IMV, L’Aigle, France) following the temperature/time curve: from 4°C to –10°C (5°C min−1), –10°C to –100°C (40°C min−1) and from –100°C to –140°C (20°C min−1), in a total of 8 min, when the straws were plunged into and stored in liquid nitrogen until evaluation. Thawed sperm (at 37°C/30 s) was diluted at 5 × 106 spermatozoa mL−1 in TALP-polyvinyl alcohol (PVA) plus Hoechst 342 (100 μg mL−1; Sigma Co., St. Louis, MO, USA). After that, samples were stained for membrane integrity with the association of fluorescent probes propidium iodide (PI, 50 μg mL−1; Sigma Co.), fluorescein thiocyanate (FITC)-Pisum sativum agglutinin (PSA, 1 mg mL−1; Sigma Co.) and Annexin V-APC (BD Pharmingen, Franklin Lakes, NJ, USA), and with MitoStatus Red (20 nM; BD Pharmingen) and YO-PRO-1 (7.5 μM; Molecular Probes Inc., Eugene, OR, USA) for MMP. Sperm samples were analysed by flow cytometer (BD LSR; Fortessa, Becton Dickinson, Mountain View, CA, USA) and the results expressed as percentage of intact cells or qualitative fluorescence expressed in arbitrary units (AU). Analysis of variance and Tukey’s test were used for statistical analysis with P < 0.05 taken as significant. There were no differences between groups for IPAM (values ranging from 45.9 ± 7.0% to 55.6 ± 8.5%). Similarly, results of PT translocation did not differ among the groups (range from 34.7 ± 7.0% to 47.6 ± 7.0%). However, there was a tendency of increasing MMP (P = 0.10) in 70UD (1789 ± 258 UA), 70D (1776 ± 162.1 UA), and 90UD (1757 ± 133.8 UA) compared with C (1368 ± 267.4 UA) and 90D (1356 ± 145 UA). In conclusion, SLC did not compromise sperm membrane functionality and it seemed to select spermatozoa with higher mitochondrial functional activity when centrifuged at the concentration of 70% and 90D. This research was funded by FAPESP # 2015/20986-3, Tairana Artificial Insemination Station, MasterFertility Ltda, Brazil.

scholarly journals Analysis of proapoptotic changes in boar spermatozoa stored at 16 °C in different short-term semen extenders

2013 ◽  
Vol 57 (3) ◽  
pp. 425-428
Author(s):  
Paweł Wysocki ◽  
Aleksandra Łyjak ◽  
Władysław Kordan
Keyword(s):  
Annexin V ◽  
Point Of View ◽  
Sperm Cells ◽  
Hoechst 33258 ◽  
Short Term ◽  
Semen Storage ◽  
Boar Semen ◽  
Boar Spermatozoa ◽  
First Time ◽  
Cell Analyzer

Abstract The aim of this study was to evaluate the effect of boar semen storage in different short-term extenders (BTS, Kortowo-3, and M III) on the percentage of spermatozoa showing proapoptotic and necrotic changes. For the first time in this study, Annexin V isolated from swine placenta has been used to determine proapoptotic changes in stored boar spermatozoa. The changes were determined using the IN Cell Analyzer 2000. A gradual decrease in motility was observed on successive days of storage. Spermatozoa incubated in the BTS extender were characterised by the highest average motility, which reached 75% on the 1st d and 39% on day 5. Motility of spermatozoa stored in BTS was significantly higher than those stored in Kortowo-3 and M III extenders after 5 d of storage. Diluted semen contained 1.5% to 2.8% spermatozoa with proapoptotic changes. The discussed process was intensified on the 3rd d of storage when the percentage of apoptotic spermatozoa was determined at 8.3% to 14.6%, and the content of dead spermatozoa exceeded 25%. The analysed extenders differed insignificantly in their ability to protect semen against proapoptotic changes during storage. From the methodological point of view, Hoechst 33258 could be used additionally to stain sperm cells regardless of their status.

61 PLANT EXTRACTS REDUCE DNA FRAGMENTATION IN FROZEN-THAWED STALLION SPERM

2009 ◽  
Vol 21 (1) ◽  
pp. 131 ◽  
Author(s):  
P. D. Burns ◽  
N. Wong ◽  
H. Arnold ◽  
N. Sirs ◽  
R. Romero ◽  
...  
Keyword(s):  
Dna Damage ◽  
Liquid Nitrogen ◽  
Dna Fragmentation ◽  
Plant Extract ◽  
Plant Extracts ◽  
Sperm Quality ◽  
Tail Length ◽  
Practical Method ◽  
Computer Assisted ◽  
Sperm Cells

Mares inseminated with frozen–thawed sperm have reduced pregnancy rates compared with mares inseminated with fresh sperm. Processing mammalian sperm for cryopreservation increases the concentration of free radicals and induces oxidative stress, which can result in DNA damage and may lead to lower fertility. The objective of this experiment was to examine the effects of several plant antioxidant extracts on stallion sperm post-thaw motility and DNA quality. Single ejaculates were collected from 4 stallions and the concentration of sperm cells in each ejaculate was determined spectrophotometrically. Semen was centrifuged at 300g for 10 min at room temperature and seminal plasma was removed. Sperm pellets were resuspended to a final concentration of 200 × 106 cells mL–1 in E-Z Freezin LE (ARS, Chino, CA) extender (control) or extender containing 1 of 3 plant extracts (3% v/v) from 2 different commercial sources. Extended sperm cells were loaded into 0.5-mL straws and frozen over liquid nitrogen vapor for 10 min. Straws were then plunged into liquid nitrogen and stored until further evaluation. Motility and velocity parameters were determined at 0, 30, and 60 min post-thaw using a computer-assisted sperm analyzer. DNA fragmentation was determined immediately after thawing using a single-cell gel electrophoresis (Comet) assay. Motility (total and progressive) and velocity parameters of sperm cells did not differ between controls and plant extract treatments (P > 0.05). However, total Comet length and tail length were reduced in sperm cells stored in extender containing each plant extract (P < 0.05). Tail and olive moment tended to be reduced (P < 0.10) in sperm cells stored in plant extracts. In conclusion, sperm cells stored in plant extracts had reduced post-thaw DNA damage. The addition of plant extracts to commercial freezing extenders may be a practical method for improving sperm quality.

scholarly journals Metabolic activity of boar semen stored in different extenders supplemented with ostrich egg yolk lipoproteins

2017 ◽  
Vol 61 (1) ◽  
pp. 127-133 ◽  
Author(s):  
Anna Dziekońska ◽  
Marek Kinder ◽  
Leyland Fraser ◽  
Jerzy Strzeżek ◽  
Władysław Kordan
Keyword(s):  
Oxygen Consumption ◽  
Metabolic Activity ◽  
Storage Temperature ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Atp Production ◽  
Beneficial Effects ◽  
Semen Storage ◽  
Boar Semen ◽  
Boar Spermatozoa

AbstractIntroduction: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.Results:The sperm metabolic activity seemed to be higher in the LPFo-based extenders following storage for three days, irrespective of the storage temperature. Compared with the LPFo-free extenders, significantly higher (P < 0.05) sperm PMI and MMP were observed in BTS and MR-A extenders supplemented with LPFo during storage for three days at 5°C. Spermatozoa stored in the BTS-LPFo extender exhibited higher (P < 0.05) TMOT and oxygen consumption, whereas higher (P < 0.05) PMI was observed in spermatozoa stored in Androhep-LPFo and MR-A-LPFo for three days at 16°C. No significant differences (P > 0.05) in ATP content were observed between the LPFo-free and LPFo-based extenders during storage.Conclusions:Supplementation of LPFo to semen extenders had varying effects on the metabolic activity of boar spermatozoa stored at different temperatures. It can be suggested that the interactions of various components of the extenders and seminal plasma with LPFo exert beneficial effects on the sperm metabolic activity during liquid storage of boar semen.

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