scholarly journals Single Layer Centrifugation Can Be Scaled-Up Further to Process up to 150 mL Semen

2011 ◽  
Vol 2011 ◽  
pp. 1-6 ◽  
Author(s):  
J. M. Morrell ◽  
M. van Wienen ◽  
M. Wallgren
Keyword(s):  
Sperm Quality ◽  
Scale Up ◽  
Single Layer ◽  
Original Technique ◽  
Glass Tubes ◽  
Boar Semen

Single-Layer centrifugation has been used to improve the quality of sperm samples in several species. However, where stallion or boar semen is to be used for AI, larger volumes of semen have to be processed than for other species, thus limiting the effectiveness of the original technique. The objective of the present study was to scale up the SLC method for both stallion and boar semen. Stallion semen could be processed in 100 mL glass tubes without a loss of sperm quality, and similarly, boar semen could be processed in 200 mL and 500 mL tubes without losing sperm quality. The results of these preliminary studies are encouraging, and larger trials are underway to evaluate using these methods in the field.

scholarly journals Single Layer Centrifugation with Androcoll-P Can Be Scaled-Up to Process Larger Volumes of Boar Semen

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
Marjet van Wienen ◽  
Anders Johannisson ◽  
Margareta Wallgren ◽  
Joyce Parlevliet ◽  
Jane M. Morrell
Keyword(s):  
Reactive Oxygen Species ◽  
Sperm Quality ◽  
Scale Up ◽  
Membrane Integrity ◽  
Single Layer ◽  
Ros Production ◽  
Oxygen Species ◽  
Normal Morphology ◽  
Preliminary Step ◽  
Boar Semen

The objective of this study was to scale-up the procedure for Single Layer Centrifugation (SLC) through AndrocollTM-P, as a preliminary step towords processing the whole ejaculate. The first experiment compared Single Layer Centrifugation using 4.5 mL and 15 mL extended ejaculate (SLC-4.5 and SLC-15, resp.), assessing sperm quality by objective motility analysis, morphology, viability, and the production of reactive oxygen species (ROS). In the second experiment, SLC-4.5 was compared to Single Layer Centrifugation with 25 mL extended ejaculate (SLC-25) using motility analysis and morphology. In both experiments, normal morphology and linear motility were significantly higher in the SLC-selected samples than in the uncentrifuged controls (P<.001), whereas total motility and membrane integrity were unchanged. Although ROS production was higher in the SLC-selected samples than in the controls (P<.01), this might have been due to the presence of antioxidants in seminal plasma in the latter. In conclusion, there was no difference in sperm quality between SLC-4.5 and SLC-15 samples, or between SLC-4.5 and SLC-25 samples, indicating that the SLC method can be scaled-up successfully.

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.

scholarly journals Efficacy of potassium permanganate and turmeric as antimicrobial agents on the bacterial load and quality of boar semen during preservation at 15 °C

2020 ◽  
Vol 90 (5) ◽  
pp. 443-451
Author(s):  
Nang Kanna Manpoong ◽  
Kutubuddin Ahmed ◽  
Dilip Kumar Bhattacharya ◽  
Dipak Kumar Sarma ◽  
Nekibuddin Ahmed ◽  
...  
Keyword(s):  
Antimicrobial Agents ◽  
Semen Quality ◽  
Sperm Quality ◽  
Bacterial Load ◽  
Conception Rate ◽  
E Coli ◽  
Bacillus Spp ◽  
Boar Semen ◽  
Conventional Antibiotics

The present study was conducted to evaluate the efficacy of different natural antimicrobials agents (KMnO4 and Turmeric) in comparison with conventional antibiotics against the bacterial load and in relation to the quality of boar semen in Modena extender for up to 120 hours of preservation at 15 °C. A total of 56 ejaculates, 14 from each of four Hampshire crossbred boars maintained within the ICAR-AICRP on Pigs, in Guwahati, Assam, India, were utilized in the study. Thirty-two ejaculates, 8 from each of four boars were used to study the effect of antimicrobial agents on semen quality during preservation at 15 °C in Modena extender. A total of 9 different bacterial types were identified from 46 bacterial isolates, obtained from 24 fresh semen samples viz. Staphylococcus aureus (24%), E. coli (22%), Bacillus spp. (13%), Citrobacter spp. (9%), Pseudomonas spp. (9%), Staphylococcus epidermidis (9%), Klebsiella spp. (6%), Streptococcus spp. (6%) and Proteus spp. (2%). The overall sensitivity of the recovered isolates to Gentamicin, Ampicillin, Enrofloxacin, Cloxacillin, Streptomycin, Penicillin, Amoxycilln, Ofloxacin and Tetracyclin were 89, 39, 37, 48, 74, 52, 56, 76 and 63% respectively. The mean sperm motility, intact acrosome, HOST-reacted spermatozoa and bacterial load differed significantly (P˂0.01) between antimicrobial agents (Gentamicin, KMnO4 and Turmeric) and preservation periods (0, 48, 72, 96 and 120 hours). Sperm quality based on Gentamicin was found to be best, followed by Turmeric and KMnO4 during preservation at 15 °C. The conception rate for the semen preserved for 0, 24, 48, 72, 96 and 120 hours of preservation was 83.33, 80.00, 75.00, 66.66, 66.66 and 50.00% respectively. In the present study, the preserved semen with ascending bacterial load containing Gentamicin did not affect the conception rate.

Colloid single-layer centrifugation improves post-thaw donkey (Equus asinus) sperm quality and is related to ejaculate freezability

2015 ◽  
Vol 27 (2) ◽  
pp. 332 ◽  
Author(s):  
I. Ortiz ◽  
J. Dorado ◽  
D. Acha ◽  
M. J. Gálvez ◽  
M. Urbano ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Single Layer ◽  
Quality Parameters ◽  
Sperm Parameters ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Equus Asinus ◽  
The Relationship ◽  
Fresh Semen

The aim of this study was to determine whether colloid single-layer centrifugation (SLC) improves post-thaw donkey sperm quality and if this potential enhancement is related to ejaculate freezability. Semen from Andalusian donkeys was frozen following a standard protocol. SLC was performed on frozen–thawed semen and post-thaw sperm parameters were compared with uncentrifuged samples. Sperm quality was estimated by integrating in a single value sperm motility (assessed by computer-assisted sperm analysis), morphology and viability (evaluated under brightfield or fluorescence microscopy). Sperm freezability was calculated as the relationship between sperm quality obtained before freezing and after thawing. Ejaculates were classified into low, medium and high freezability groups using the 25th and 75th percentiles as thresholds. All sperm parameters were significantly (P < 0.01) higher in SLC-selected samples in comparison to uncentrifuged frozen–thawed semen and several kinematic parameters were even higher than those obtained in fresh semen. The increment of sperm parameters after SLC selection was correlated with ejaculate freezability, obtaining the highest values after SLC in semen samples with low freezability. We concluded that, based on the sperm-quality parameters evaluated, SLC can be a suitable procedure to improve post-thaw sperm quality of cryopreserved donkey semen, in particular for those ejaculates with low freezability.

scholarly journals A new sperm selection criterion for cryopreservation of boar semen

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Monika Trzcińska ◽  
Magdalena Bryła
Keyword(s):  
Sperm Quality ◽  
Selection Criterion ◽  
Motile Sperm ◽  
Before And After ◽  
Viable Sperm ◽  
Sperm Membrane ◽  
Boar Semen ◽  
Acrosome Integrity ◽  
Fresh Semen

AbstractThis study aimed to define potential markers that could determine the suitability of ejaculate for cryopreservation. Fresh semen from eleven boars (4–7 ejaculates/boar), regardless of their sperm motility, was subjected to a cryopreservation procedure. The sperm quality before and after freezing was assessed based on the sperm membrane permeability and acrosome integrity. The results showed that it was possible to effectively cryopreserve ejaculates below the accepted standards of 70–80% of fresh motile sperm and still obtain a high cryosurvival rate. Moreover, a significant correlation was found between the percentage of viable sperm with apoptotic-like changes, viable sperm with reacted acrosomes, and the cryosurvival rate. The proposed markers for assessing the quality of fresh semen could be used to predict the success of cryopreservation procedures.

Single layer centrifugation (SLC) improves sperm quality of cryopreserved Blanca-Celtibérica buck semen

2012 ◽  
Vol 136 (1-2) ◽  
pp. 47-54 ◽  
Author(s):  
P. Jiménez-Rabadán ◽  
J.M. Morrell ◽  
A. Johannisson ◽  
M. Ramón ◽  
O. García-Álvarez ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Single Layer

Does single‐layer centrifugation with Bovicoll improve sperm quality of frozen‐thawed semen in Fleckvieh bulls?

2020 ◽  
Vol 55 (6) ◽  
pp. 691-700
Author(s):  
Joanna Szlendak ◽  
Christine Adler ◽  
Jakob Scherzer ◽  
Anna Niwinska ◽  
Ewa Kautz ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Single Layer

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.

Hydroxytyrosol effectively improves the quality of pig sperm at room temperature

2020 ◽  
Author(s):  
Wenuyu Zhang ◽  
Dong Li ◽  
Xueka Tian ◽  
Yulin He ◽  
Zitong Xiao ◽  
...  
Keyword(s):  
Artificial Insemination ◽  
Room Temperature ◽  
Sperm Quality ◽  
Membrane Damage ◽  
Treated Group ◽  
Control Group ◽  
Sequencing Analysis ◽  
Oxygen Species ◽  
Boar Semen

Abstract Background: Artificial insemination technology is the production technology used in more than 99% of the world's pig farms. Preservation of semen is mainly conducted at room temperature (17°C), during which the accumulation of reactive oxygen species (ROS). In our research, by adding different concentrations of hydroxytyrosol to the diluent during the storage of boar semen at 17°C, we compared them with the traditional diluent to study the cumulative amount of ROS, the extent of membrane damage, the sperm acrosome integrity, the activity of antioxidant enzymes, and other indicators to evaluate the effect of hydroxytyrosol on the changes of active oxygen and sperm quality during storage. After we determined the optimal amount of hydroxytyrosol in the diluent, Proteomics was used to monitor difference in sperm protein expression between the control group and the addition of 120 μmol/L hydroxytyrosol (optimum concentration) group after three days’ storage. Ultimately, we selected the control group and the addition of 120 μmol/L hydroxytyrosol group to test the effect of artificial insemination.Results: During storage of pig semen at room temperature, the sperm quality of the hydroxytyrosol-treated group was significantly improved (P < 0.05). Besides, its antioxidant capacity was also significantly improved (P < 0.05). Proteomics sequencing analysis proved that the addition of hydroxytyrosol treatment group has potential value in improving sperm cell quality. The significant increase in sow pregnancy rate and the increase in piglet birth weight also confirmed that sperm quality during storage can be of vital importance in actual production (P < 0.05).Conclusions: Based on our results, we demonstrated that the addition of hydroxytyrosol to the diluent can not only improve the quality of pig sperm, but also has an irreplaceable role in improving the efficiency of artificial insemination.

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