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.

130 EFFECTIVENESS OF BUTYLATED HYDROXYTOLUENE AS EGG YOLK SUBSTITUTE FOR CRYOPRESERVATION OF BOAR SPERMATOZOA

2007 ◽  
Vol 19 (1) ◽  
pp. 182 ◽  
Author(s):  
L. Rodriguez-Vilar ◽  
M. Hernandez ◽  
C. Lopez-Sanchez ◽  
J. M. Vazquez ◽  
E. A. Martinez ◽  
...  
Keyword(s):  
Protective Effect ◽  
Mixed Model ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Positive Control ◽  
Negative Control ◽  
Butylated Hydroxytoluene ◽  
Main Effects ◽  
Boar Spermatozoa

Butylated hydroxytoluene (BHT) has proven to be efficient as a supplement for cryopreservation boar spermatozoa (Roca et al. 2004 J. Androl. 25, 397–405). Moreover, it has been successfully used as an egg yolk substitute to cryopreserve goat spermatozoa (Khalifa and El-Saidy 2006 Anim. Reprod. Sci. 93, 303–315). The objective of this study was to evaluate the effectiveness of BHT as an egg yolk substitute for freezing boar spermatozoa. Nine sperm-rich ejaculate fractions were collected from 3 boars (3 ejaculates per boar) using the gloved-hand method. After centrifugation (2400g for 3 min), the sperm pellet of each ejaculate was split into 5 aliquots. The aliquots were diluted (to a final concentration of 1 � 109 sperm/mL) in a Tris-citric-glucose extender with 3% glycerol and supplemented with 20% egg yolk (positive control, PC aliquot) or BHT at the final concentrations of 0 (negative control, NC aliquot), 0.2, 0.4, and 0.8 mM. Diluted semen samples were dispensed into 0.5-mL straws, and frozen in a programmable cell freezer at 20�C min. Thawing was carried out in a water bath at 70�C for 8 s. Post-thaw sperm survival was assessed according to total sperm motility (TSM, %) using a CASA system (SCA�; Microptic, Barcelona, Spain), and plasma membrane integrity (PMI, %) and acrosome membrane integrity (AMI, %) using a flow cytometric procedure (SYBR-14/propidioum iodide/FITC-phycoerythrin), at 30 and 150 min post-thawing in diluted Beltsville thawing solution with spermatozoa held in a waterbath at 37�C (3 straws per ejaculate). Data were analyzed using a ANOVA mixed model including the main effects of aliquot, boar, post-thaw assessment time, and their interactions, with ejaculate and straw as random effects. All main effects had significant influence (P ≤ 0.01) in all post-thaw sperm assessments. However, no interactions (P ≥ 0.05) among main effects were shown. Data were combined for the 2 post-thaw assessment times. The best (P ≤ 0.05) post-thaw sperm quality (mean � SEM) was achieved in PC aliquots (47.11 � 3.10, 58.98 � 2.78, and 51.35 � 3.42 for TSM, PMI, and AMI, respectively). In NC aliquots, the percentage of TSM, PMI, and AMI were always below 1% (P ≤ 0.05). BHT has a beneficial (P ≤ 0.05) effect on post-thaw sperm assessments, and no differences (P ≥ 0.05) among concentrations were shown. The mean post-thaw sperm quality in the BHT aliquots was 8.50 � 0.80, 20.29 � 0.53, and 16.03 � 0.55 for TSM, PMI, and AMI, respectively. On the basis of these data, we can conclude that BHT has a protective effect for boar spermatozoa during the cryopreservation process. However, BHT alone is insufficient to replace the protective effect of egg yolk. This work was supported by CICYT (AGF2005-00706), Madrid, Spain.

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 Effects of Tris (hydroxymethyl) aminomethane on the quality of frozen-thawed boar spermatozoa

2019 ◽  
Vol 67 (1) ◽  
pp. 106-114
Author(s):  
Zhao Namula ◽  
Fuminori Tanihara ◽  
Manita Wittayarat ◽  
Maki Hirata ◽  
Nhien Thi Nguyen ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Control Group ◽  
Freezing And Thawing ◽  
Control Groups ◽  
Boar Semen ◽  
Boar Spermatozoa ◽  
Time Point

Tris (hydroxymethyl) aminomethane (Tris) has been used as a pH regulator for buffering the pH of dilution extenders for boar semen, such as the Modena extender. The purpose of the present study was to assess the effects of Tris supplementation at different concentrations (0, 8, 24 and 72 μM) into the freezing extender on the quality and fertilising capacity of frozen-thawed boar spermatozoa. The results showed that the supplementation of 24 μM of Tris gave significantly higher percentages of sperm viability and plasma membrane integrity than those of the control group at any time point of assessment (0 h and 3 h post-thawing) (P < 0.05). However, there were no significant differences in the acrosome integrity parameter among the groups. Higher percentages of sperm motility were observed in the spermatozoa cryopreserved with 24 μM of Tris compared to the control groups when the samples were analysed 0 h after thawing (P < 0.05). However, an increase of the Tris concentration to 72 μM did not enhance the sperm motility parameters. The total numbers of fertilised oocytes and blastocysts obtained with spermatozoa frozen with 24 μM Tris were significantly higher than those of the control group without Tris (P < 0.05). In conclusion, the supplementation of 24 μM Tris into the freezing extender contributes to a better boar sperm quality and fertilising capacity after the process of freezing and thawing.

scholarly journals THE PRODUCTION OF FREEZE-DRIED EGG YOLK POWDER AND ITS EFFECT ON THE QUALITY OF GARUT RAM LIQUID SEMEN

2021 ◽  
Vol 15 (2) ◽  
Author(s):  
Oriza Savitri Ariantie ◽  
Amrozi Amrozi ◽  
Tuty Laswardi Yusuf ◽  
Nurul Taufiqu Rochman ◽  
Bambang Purwantara
Keyword(s):  
Particle Size ◽  
Sperm Quality ◽  
Egg Yolk ◽  
Progressive Motility ◽  
Freeze Dried ◽  
Semen Preservation ◽  
Viability Percentage ◽  
Index Value ◽  
Ram Sperm

This research aims to evaluate the protective potency of freeze-dried egg yolk powder on Garut ram sperm during the liquid semen preservation process. Semen with good sperm quality was divided into three groups and diluted using the following Tris diluents: fresh egg yolk-Tris (FEY-Tris), commercial egg yolk powder-Tris (CEY-Tris), and freeze-dried egg yolk powder-Tris (DEY-Tris). Semen that had been diluted was observed every 12 hours until sperm progressive motility was 50%. Sperm quality was tested for progressive motility percentages, viability, intact plasma membrane (IPM), and intact acrosome (IA). Fresh egg yolk had a particle size of 14460.00±330.76 nm, with polydispersity index value (PI) and zeta potential (ZP) being 1.00±0.11 and -9.22±0.30 mV, respectively. The particle size of commercial egg yolk powder was 877.90±168.86 nm with PI value 0.34±0.04 and ZP-28.7±1.24 mV. The particle size of freeze-dried egg yolk powder was 1296.00±86.73 nm, with PI value 0.70±0.04 and ZP -34.5±0.64 mV. Progressive motility percentage showed that DEY-Tris diluent managed to survive to 51.11±6.06% for 168 hours, compared to FEY-Tris (51.25±6.74%) which survived for 156 hours, although no longer than CEY-Tris diluent (53.47±6.33%) which survive for 180 hours (P0.05). Higher sperm motility was also supported by viability percentage, IPM, and IA, which were all higher (P0.05). In conclusion, freeze dried egg yolk powder mixed with Tris buffer was able to maintain sperm quality during the liquid semen preservation process.

scholarly journals Assessment of quality in specific fractions of Large White Yorkshire boar semen

2021 ◽  
Vol 52 (2) ◽  
Author(s):  
K. G. Ambily ◽  
Malati Naik ◽  
Hiron M. Harshan ◽  
C. Jayakumar ◽  
M. P. Unnikrishnan ◽  
...  
Keyword(s):  
Cold Shock ◽  
Sperm Concentration ◽  
Membrane Integrity ◽  
Membrane Cholesterol ◽  
Large White ◽  
Progressive Motility ◽  
Sperm Membrane ◽  
Boar Semen ◽  
Quality Assessments

Boar semen is voluminous and ejaculated as jets or fractions of pre-sperm, sperm rich (SRF) and post-sperm rich fractions. Recent studies have reported more resilient characteristics of sperm in initial portions of SRF towards cold shock and cryopreservation. The present study was conducted to assess the quality of specific fractions of SRF, namely, first 10mL of SRF (F1) and rest of SRF (F2) in Large white Yorkshire (LWY) boar semen. Ejaculates were collected using gloved-hand technique and were subjected to quality assessments of volume, pH, sperm progressive motility, concentration, plasma membrane integrity, abnormality, acrosome integrity and sperm membrane cholesterol. Upon statistical analysis, significant differences were noticed in volume, pH, sperm concentration and sperm membrane cholesterol between fractions of the ejaculate.

P–025 Sperm selection using a modified “swim up” technique in absence of sperm centrifugation improve sperm DNA fragmentation and decreases miscarriage rate

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Y Cabell. Vives ◽  
P Belchin ◽  
C Lopez-Fernandez ◽  
M Fernandez-Rubio ◽  
J Guerrero-Sanchez ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Sperm Quality ◽  
P Value ◽  
Sperm Dna Fragmentation ◽  
Sperm Selection ◽  
Male Factor ◽  
Miscarriage Rate ◽  
Sperm Dna ◽  
Group 2

Abstract Study question Is it useful to avoid sperm centrifugation in laboratory routine work to improve sperm quality and reproductive outcome in Assisted Reproduction Techniques (ART)? Summary answer Exclusion of sperm centrifugation for sperm selection using neat sperm samples (IO-lix), increases sperm quality in the collected subpopulation decreasing miscarriage rate after using ICSI. What is known already Inclusion of sperm centrifugation in ART is an aggressive intervention for sperm selection with ineludible production iatrogenic damage affecting sperm integrity. The application of IMSI, PICSI or microfluidic devices avoid sperm centrifugation and may improve the quality of the subsample obtained. However, these methodologies may result time consuming, expensive or producing poor results when the quality of the sperm is limited. We have already shown that a modified swim-up avoiding centrifugation (called IO-lix) is a low-cost and efficient alternative to microfluidic devices, recovers 100 times more concentration and reduces sperm DNA fragmentation with no significant differences to other methodologies. Study design, size, duration This is a retrospective study from 2018 to 2020 which includes patients with an average of age of 38.2 years using their own oocytes with ICSI as fertilization technique. Two aleatory groups of patients were made: Group 1: 88 cycles with 503 fertilized oocytes and 206 blastocysts were obtained with sperm samples processed by IO-lix and Group 2: 303 cycles, 1451 fertilized oocytes and 591 blastocysts using a standard “swim up” technique to process sperm. Participants/materials, setting, methods A total of 391 ICSI cycles were included in this retrospective study. The male factor was similar in both groups and they showed altered SDF previously to the cycle. We compared data of the motility and SDF of sperm samples before and after applying IO-lix and we analyzed by X2 contingence test differences on miscarriage rates between groups 1 and 2. Main results and the role of chance General sperm parameter changes after IO-lix showed that averaged sperm concentration observed in neat ejaculated samples was 62M/SD=46.4. Values obtained after IO-lix in the same samples were 12.3M/SD8.0. Averaged sperm motility in neat samples was 54%/SD=9.3 and 70.9%/SD=13.2 after IO-lix. Finally, sperm DNA fragmentation in neat samples was 35.8%/SD17.3, while these values decreased to 9.2%/SD=3.9 after IO-lix. About reproductive outcome results, significant differences were not obtained on the development to blastocyst stage rate comparing both groups (X2=0.003; p value = 0.954; Alpha 0.05). In the case of IO-lix processed samples, the pregnancy rate was 59.42% in Group 1 and 44.72% in Group 2 (X2=0.651; p value =0.419; Alpha 0.05). A total of 9 miscarriages of 41 clinical pregnancies (21.95%) were observed after IO-lix, while this number increases to 59 out of 123 clinical pregnancies, which means the 47.96% of the embryo transfers, when “swim-up” was used. In this case significant differences were obtained (X2=3.935; p value = 0.0.047; Alpha 0.05). Limitations, reasons for caution Being a pilot study aimed to understand the results of IO-lix in ART, correlations have not been stablished between the levels of sperm improvement after IO-lix and paired results of ART. This study would be necessary, specially to identify the possible origin of miscarriage associated to the male factor. Wider implications of the findings: Elimination of sperm centrifugation using a combined strategy of gradients and “swim-up” for sperm isolation, reduce miscarriage rate and produce equivalent results of blastocyst development to those obtained with “swim-up”. Being a cost-effective and improving laboratory workload, its use for sperm selection is recommended. Trial registration number Not applicable

COMPARATIVE CHARACTERISTICS OF RAM SEMEN FROZEN IN DIFFERENT EXTENDERS

2021 ◽  
pp. 63-78
Author(s):  
M.M. AYBAZOV ◽  
◽  
A.N. SHEVCHENKO ◽  
M.I. SELIONOVA ◽  
T.V. MAMONTOVA
Keyword(s):  
Egg Yolk ◽  
Quality Parameters ◽  
Semen Parameters ◽  
Progressive Motility ◽  
Kinematic Performance ◽  
Synthetic Media ◽  
Ram Sperm

Numerous studies have proved the necessity of egg yolk in synthetic media to dilute semen before its cryopreservation. However, at the same time, it has been demonstrated that its use can adversely affect the quality of frozen-thawed sperm. The present study compares the main quality parameters of ram sperm frozen using TRIS-based diluent with egg yolk and two egg yolk-free diluents (OvixCell® and AndroMed®). A slower deterioration in the kinematic performance of sperm cryopreserved in TRIS diluent with native egg yolk confirmed higher cryoprotective performance compared to commercial extenders containing no egg yolk. Significantly higher total and progressive motility was observed in TRIS-based medium with egg yolk (P< 0.05). This advantage was maintained after four hours of incubation and became more significant at the end of cultivation (after six hours) (P< 0.01). Thus, ram sperm frozen in egg yolk medium retained better motility than in egg yolk-free extenders, which allows predicting its higher bioavailability. Assessment of some semen parameters using CASA showed that there were no significant differences in motility between the three extenders immediately after thawing the straws. When assessed two hours after thawing, a diluent containing egg yolk (TRIS-based) was found to have higher results for some of the examined traits than phospholipid diluents.

Value of semen parameters, with special reference to TNF-α, in predicting the quality of boar semen after short-term storage

2013 ◽  
Vol 61 (2) ◽  
pp. 209-219 ◽  
Author(s):  
Janko Mrkun ◽  
Marjan Kosec ◽  
Petra Zrimšek
Keyword(s):  
Semen Quality ◽  
Semen Parameters ◽  
Short Term ◽  
Progressive Motility ◽  
Tnf Α ◽  
Semen Storage ◽  
Boar Semen ◽  
Α Level ◽  
Term Storage

The aim of this study was to address the question whether changes in boar semen quality after short-term storage could be predicted on the basis of standard semen parameters and TNF-α level determined on the day of semen collection under commercial conditions. Progressive motility showed the highest positive correlation with morphology on day 0 of collection, and progressive motility on day 3 (P < 0.05) showed a negative correlation with acrosome abnormalities (P < 0.05). According to the area under receiver operating characteristics (ROC) curves (AUCs), progressive motility could also be used in predicting semen quality after 3 days of storage (AUC > 0.5; P < 0.05). TNF-α in seminal plasma is the only parameter measured on day 0 to show a significant correlation with the percentage of viable spermatozoa after 3 days of semen storage (r = 0.495, P < 0.05). ROC analysis shows that TNF-α level is helpful in discriminating viability outcome after semen storage (AUC = 0.94, P < 0.001). We can predict with 92.35% certainty that fresh semen samples with more than 150 pg/ml of TNF-α in the seminal plasma will retain more than 85% of viable spermatozoa after 3 days of storage. Thus, TNF-α can contribute to predicting the quality of short-term stored semen.

105 Increasing fertility of interspecific hybrid males using biotechnological approaches

2021 ◽  
Vol 33 (2) ◽  
pp. 159
Author(s):  
A. Vetokh ◽  
A. Tadzhieva ◽  
B. Iolchiev ◽  
N. Volkova ◽  
V. Bagirov
Keyword(s):  
Dna Fragmentation ◽  
Interspecific Hybrid ◽  
Semen Quality ◽  
Sperm Quality ◽  
Semen Analysis ◽  
Differential Staining ◽  
Computer Assisted ◽  
Sperm Dna Fragmentation ◽  
Russian Science

The results of AI depend on many factors, with the quality of semen being one of the most important. Not all male hybrids can meet the requirements for semen quality, because they often have reduced fertility following cryopreservation. Thus, it is necessary to improve semen processing before use in AI. The aim of the study was to evaluate the effectiveness of using the “swim-up” flotation method to improve sperm quality of hybrid males of the Ovis genus. Semen from interspecific hybrid rams (1/4 Argali×3/4 Romanov, n=15; 1/8 Argali×7/8 Romanov, n=15) was freshly obtained, frozen–thawed, and processed by the swim-up method. Evaluation of sperm motility was determined using computer-assisted semen analysis. Statistical analysis was performed using SPSS vs.15.0 (ANOVA and t-test; SPSS Inc.). Semen was collected during the breeding season (October–December) via artificial vagina. Assessment of acrosome integrity was determined using differential staining with a Diachem diff-quick kit (NPF ABRIS+). The degree of sperm DNA fragmentation was determined using the acridine-orange test. The sperm freezing/thawing cycle was accompanied by sperm damage and an increase in the proportion of immobile sperm from 10 to 58%, with non-progressive movement increasing from 9 to 19.3%. The number of spermatozoa with abnormal morphology doubled, and the DNA fragmentation index increased from 16 to 26%. Use of the swim-up procedure allowed us to sort progressively motile spermatozoa. The content of progressively motile spermatozoa in the samples obtained from the supernatant was 86%, which was 2.3 times higher than in frozen–thawed sperm (P≤0,01). The obtained results show the effective use of the swim-up procedure to determine the quality of semen in hybrid rams. These studies were carried out with financial support from the Russian Science Foundation, grant No. 18-16-00079 and the Ministry of Science and Higher Education of the Russian Federation.

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.

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