Relevance of the Ejaculate Fraction and Dilution Method on Boar Sperm Quality during Processing and Conservation of Seminal Doses

During boar semen processing and distribution, maximizing the work protocols in the laboratories becomes essential for the conservation of seminal doses. One of the recent implementations in the boar studs to improve efficiency has been semi-automatic semen collection systems, which do not allow to discard fractions of the ejaculate. The objective of this work was to evaluate the dilution method and vibrations (simulating delivery transport) effect on sperm quality (motility, viability, morphology, thermo-resistance test) according to the fraction of ejaculate collected. Two different fractions of the ejaculate were obtained [rich fraction (RF); total fractions (TF)] from six boars, and two dilution methods applied [pouring the extender over the semen (control; ES); pouring the semen over the extender (reverse; SE)]. The seminal doses (2000 × 106 sperm/50 mL) were preserved for 5 days. The results showed that the fraction collected affects sperm quality (better total and progressive motility, and faster sperm in TF; p < 0.05) regardless of the dilution method applied. However, these differences diminished after submitting the semen to the thermo-resistance test, with only differences in sperm viability being observed (p < 0.05). When seminal doses were subjected to vibrations, the sperm viability was more affected in the TF than in the RF group (p < 0.05). In conclusion, using the TF ejaculate leads to comparable results to the RF in sperm quality during storage regardless of the dilution method applied. However, the vibrations of seminal doses are more affected in doses prepared with TF than with RF, although more factors should be included to approach the real conditions during transport.

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The use of butylated hydroxytoluene in cryopreservation of boar semen

Roczniki Naukowe Polskiego Towarzystwa Zootechnicznego ◽

10.5604/01.3001.0013.6966 ◽

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.

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Changes in sperm quality and lipid composition during cryopreservation of boar semen

Reproduction ◽

10.1530/rep.0.1210395 ◽

2001 ◽

pp. 395-401 ◽

Cited By ~ 125

Author(s):

S Cerolini ◽

A Maldjian ◽

F Pizzi ◽

TM Gliozzi

Keyword(s):

Lipid Content ◽

Free Cholesterol ◽

Sperm Quality ◽

Cholesterol Content ◽

Sperm Viability ◽

Freezing And Thawing ◽

Motile Cells ◽

Boar Semen ◽

Thawing Procedure ◽

Fresh Semen

The effect of cryopreservation on boar sperm viability, motility, lipid content and antioxidant enzymatic activities was studied. Three classes of semen were determined according to a cluster analysis on the basis of the proportion of live and dead cells after freezing and thawing. The classes identified were: high (H, n = 4), average (A, n = 12) and low (L, n = 3) viability. The concentration of sperm cells decreased from class H to A to L. Fresh semen samples with higher viability and a higher proportion of motile cells also maintained better quality after the freezing and thawing procedure. Sperm viability and motility in both fresh and thawed samples were similar in classes H and A, while significantly lower values were measured in class L. The relative decrease in sperm viability and motility after cryopreservation increased from class H to A to L. The lipid content of spermatozoa (micrograms per 10(9) cells) increased significantly after freezing and thawing in classes H and A but not in class L. This result indicated that active sperm lipid metabolism might be responsible for the increase in lipid content. Phospholipid and triacylglycerol contents increased whereas free cholesterol content decreased after thawing. The fatty acid composition of fresh spermatozoa was similar in all three classes. The proportion of polyunsaturated fatty acids decreased significantly after freezing and thawing, indicating contamination from the diluent or peroxidation. After freezing and thawing, superoxide dismutase activity in spermatozoa was significantly higher in class L than in classes H and A, which did not differ from each other.

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The Antibacterial and Antioxidant Roles of Buckwheat Honey (BH) in Liquid Preservation of Boar Semen

BioMed Research International ◽

10.1155/2021/5573237 ◽

2021 ◽

Vol 2021 ◽

pp. 1-9

Author(s):

Qun Lan ◽

Yingyu Xie ◽

Jiahua Pan ◽

Qiaohui Chen ◽

Tianfang Xiao ◽

...

Keyword(s):

Sperm Quality ◽

Antioxidant Properties ◽

Membrane Integrity ◽

Storage Period ◽

Antibiotic Use ◽

Quality Parameters ◽

Progressive Motility ◽

Liquid Storage ◽

Boar Semen

In the present study, we hypothesized that buckwheat honey (BH) should be regarded as a potential alternative to antibacterial and antioxidant agent in liquid storage of boar semen. To this end, boar semen was firstly studied for in vitro dose tolerability to BH by measuring sperm progressive motility. The optimum progressive motility of boar spermatozoa was observed in extender with 0.5% and 0.6% BH addition. Afterward, sperm quality parameters, bacterial profile and composition, total antioxidant (T-AOC), catalase (CAT), superoxide dismutase (SOD), and malondialdehyde (MDA) levels of control, BH supplementation, antibiotics supplementation, and incorporated supplementation were compared during liquid storage period, to further investigate antibacterial and antioxidant properties of BH. The results showed that BH supplementation significantly improved sperm motility, acrosome integrity, plasma membrane integrity, inhibited opportunistic bacterial growth, and altered microbial compositions at the end of preservation. Additionally, T-AOC, SOD, and CAT levels were significantly higher in the BH supplementation group than those in the control and antibiotic supplementation group, whereas MDA level exhibited opposite change pattern. Importantly, BH addition to the extender was able to exert a synergistic effect in combination of antibiotic use. Our findings suggested that the appropriate concentrations (0.5% and 0.6%) of BH were added to the extender could act antibacterial and antioxidant roles in liquid preservation of boar semen.

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Effect of temperature and time after collection on buck sperm quality

BMC Veterinary Research ◽

10.1186/s12917-019-2135-y ◽

2019 ◽

Vol 15 (1) ◽

Author(s):

Kirsten Hahn ◽

Klaus Failing ◽

Axel Wehrend

Keyword(s):

Semen Quality ◽

Sperm Quality ◽

Semen Analysis ◽

Effect Of Temperature ◽

Sperm Viability ◽

Factors Affecting ◽

Semen Collection ◽

Examination Time ◽

Collection Period ◽

Time Point

Abstract Background Different parameters are assessed as part of the semen analysis but a standard protocol for evaluation of goat semen is still missing. The aim of this study was to analyse two different factors affecting buck sperm quality in the post-collection period prior to adding the extender. Here we examined the effects of two handling temperatures (20 °C, 37 °C) and various examination time points (3–30 min) after semen collection. Results Examination time point had a significant influence on raw sperm viability (p < 0.05), motility (p < 0.05) and on semen pH (p < 0.05). The two different handling temperatures had no significant effect on sperm viability (p > 0.05), motility (p > 0.05), with the exception of fast moving sperm (p = 0.04), or on semen pH (p > 0.05). Conclusion Examination time point was identified as factor strongly influencing raw peacock buck semen after collection. Raw goat semen can tolerate room temperatures for at least 10 min without impacting overall semen quality. In order to obtain comparable results, semen samples should always be examined within 10 min after collection.

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52 Difference of Seminal Plasma Proteins in Good- and Poor-Freezability Boar Ejaculates

Reproduction Fertility and Development ◽

10.1071/rdv30n1ab52 ◽

2018 ◽

Vol 30 (1) ◽

pp. 165 ◽

Cited By ~ 2

Author(s):

J. Rungruangsak ◽

J. Suwimonteerabutr ◽

S. Asawakarn ◽

K. Buranaamnuay ◽

N. Chantaravisoot ◽

...

Keyword(s):

Seminal Plasma ◽

Plasma Proteins ◽

Protein Extraction ◽

Normal Morphology ◽

Progressive Motility ◽

Semen Collection ◽

Seminal Plasma Proteins ◽

Boar Semen ◽

One Way Anova ◽

Sperm Functions

Seminal plasma is the semen components that maintain sperm metabolism, pH and osmolality. Fibronectin (FN1) and glutathione peroxidase (GPX5) are the seminal plasma proteins that play an important role on the boar sperm functions. The purpose of the present study was to determine the differences in GPX5 and FN1 contents in the boar semen having good, moderate, and poor freezability. A total of 38 ejaculates from 25 boars with proven fertility were included in the experiment. All the ejaculates included in the study had >70% subjective motility, >75% normal morphology, >75% sperm viability, and volume >100 mL per ejaculate. The semen was collected through semen collection bag with filter and split into 2 portions. The first portion was prepared for the evaluation of seminal plasma proteins (i.e. GPX5 and FN1) and the second portion was cryopreserved and evaluated for post-thaw semen qualities. The seminal plasma samples were collected in cryotube and plug into liquid nitrogen. The samples was stored at –80°C before protein extraction. After thawing, the ejaculates were classified into 3 groups according to their post-thawed sperm motility: poor (14.6 ± 3.9%), modersate (28.5 ± 4.1%), and good (64.0 ± 8.7%) freezability. The amounts of GPX5 and FN1 proteins were evaluated through Western blot analysis. The normalized quantity of proteins was compared among groups by one-way ANOVA. The normalized level of FN1 in seminal plasma was higher in good- than in poor-freezability groups (8.0 ± 0.8% v. 5.7 ± 0.7%, respectively; P < 0.05), but did not differ significantly compared with that of the moderate-freezability group (7.5 ± 0.8%; P > 0.05). The levels of GPX5 in good-, moderate-, and poor-freezability groups were 14.7 ± 3.0%, 16.8 ± 3.2%, and 10.9 ± 3.0%, respectively (P > 0.05). The level of FN1 in seminal plasma was significantly correlated with the post-thaw sperm progressive motility (r = 0.38, P = 0.01), total motility (r = 0.37, P = 0.02), and the proportion of bent tail sperm (r = –0.33, P = 0.04). The level of GPX5 was not correlated with any of the post-thaw sperm qualities (P > 0.05). However, the levels of GPX5 was positively correlated with FN1 (r = 0.40, P = 0.01, n = 38). It can be concluded that FN1 in seminal plasma can be used as a marker of sperm freezability in boar.

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Proline Protects Boar Sperm against Oxidative Stress through Proline Dehydrogenase-Mediated Metabolism and the Amine Structure of Pyrrolidine

Animals ◽

10.3390/ani10091549 ◽

2020 ◽

Vol 10 (9) ◽

pp. 1549

Author(s):

Chengwen Feng ◽

Zhendong Zhu ◽

Wenjing Bai ◽

Rongnan Li ◽

Yi Zheng ◽

...

Keyword(s):

Oxidative Stress ◽

Sperm Quality ◽

Redox Homeostasis ◽

Membrane Integrity ◽

Underlying Mechanism ◽

Proline Dehydrogenase ◽

Progressive Motility ◽

Boar Sperm ◽

Amine Structure ◽

Boar Semen

Proline was reported to improve sperm quality in rams, stallions, cynomolgus monkeys, donkeys, and canines during cryopreservation. However, the underlying mechanism remains unclear. The aim of this study was to investigate the effect of proline on boar semen during liquid storage at 17 °C and explore the underlying mechanism. Freshly ejaculated boar semen was supplemented with different concentrations of proline (0, 25, 50, 75, 100, 125 mM) and stored at 17 °C for nine days. Sperm motility patterns, membrane integrity, ATP (adenosine triphosphate), reactive oxygen species (ROS), and GSH (glutathione) levels, and the activities of catalase (CAT) and superoxide dismutase (SOD) were evaluated after storage for up to five days. It was observed that boar sperm quality gradually decreased with the extension of storage time, while the ROS levels increased. Addition of 75 mM proline not only significantly improved sperm membrane integrity, motility, and ATP levels but also maintained the redox homeostasis via increasing the GSH levels and activities of CAT and SOD. When hydrogen peroxide (H2O2) was used to induce oxidative stress, addition of proline significantly improved sperm quality and reduced ROS levels. Moreover, addition of proline also improved sperm quality during the rapid cooling process. Notably, addition of DL-PCA (DL-pipecolinic acid) rescued the reduction of progressive motility and total motility caused by H2O2, and THFA (tetrahydro-2-furoic acid) failed to provide protection. Furthermore, addition of proline at 75 mM increased the activity of proline dehydrogenase (PRODH) and attenuated the H2O2-induced reduction in progressive motility. These data demonstrate that proline protects sperm against oxidative stress through the secondary amine structure and proline dehydrogenase-mediated metabolism.

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Effect of genistein addition to equine sperm freezing extender

Journal of the Hellenic Veterinary Medical Society ◽

10.12681/jhvms.15868 ◽

2018 ◽

Vol 66 (4) ◽

pp. 241 ◽

Cited By ~ 3

Author(s):

Β. MACÍAS-GARCÍA ◽

Τ. GUIMARÃES ◽

G. LOPES ◽

A. ROCHA ◽

L. GONZÁLEZ-FERNÁNDEZ

Keyword(s):

Tyrosine Phosphorylation ◽

Protein Tyrosine Kinase ◽

Kinase Inhibitor ◽

Sperm Quality ◽

Protein Tyrosine Phosphorylation ◽

Sperm Viability ◽

Protein Tyrosine ◽

Progressive Motility ◽

Ros Scavenger

Cryopreservation negatively affects equine sperm quality post-thaw: an excessive release of reactive oxygen species (ROS) and increased protein tyrosine phosphorylation (known as cryocapacitation) have been demonstrated in frozenthawed equine sperm diminishing its lifespan. The objective of this study was to determine the possible beneficial effect of genistein addition (a ROS scavenger and protein tyrosine kinase inhibitor) to a commercial freezing extender. Equine sperm were frozen in the presence of different genistein concentrations ranging from 0 to 800 μM. After thawing, the sperm viability (eosinnigrosin), acrosome integrity using peanut agglutinin (PNA), protein tyrosine phosphorylation (PY) and motility were assessed at times 0 and 1 hr. In addition PY was studied in fresh and frozen sperm incubated in Modified Whitten’s (MW) medium with 25 mM Bicarbonate (MW+Bic). Genistein did not affect the viability, total or progressive motility or acrosome status of frozenthawed equine sperm. Immediately after thawing, positive PY staining in the equatorial band was observed and genistein addition did not exert any change in PY. Fresh sperm incubated in MW+Bic showed a significant increase in PY staining along the tail compared to frozen-thawed sperm. In our study sperm viability immediately post-thaw was 58.25% ± 5.35 and progressive motility 14.46% ± 5.7 (mean ± S.E.M.) and genistein did not improve the post-thaw quality of equine sperm. In addition, cryopreservation did not induce PY immediately post-thaw or enhanced frozen-thawed equine sperm susceptibility to PY induction.

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Effect of warmed semen extender on boar sperm quality post-collection

Transactions of the Missouri Academy of Science ◽

10.30956/mas-30 ◽

2019 ◽

Vol 47 (2019) ◽

pp. 13-17

Author(s):

K.W. Lovercamp ◽

A. Giri

Keyword(s):

Sperm Quality ◽

Negative Effects ◽

Swine Industry ◽

Semen Collection ◽

Boar Sperm ◽

Boar Semen ◽

Semen Extender ◽

And Control ◽

Sexually Mature

Abstract Semen used for artificial insemination (AI) in the swine industry is typically collected into a warmed semen collection cup containing an empty collection bag. If the ambient temperature does not closely match the temperature of the warmed collection cup and semen at the time of collection then negative effects to the motility and morphology of the sperm cells may occur due to temperature shock. The purpose of this research was to determine if collecting boar semen directly into semen extender warmed to 38.5°C would affect sperm quality post-collection. Sexually mature Berkshire x Duroc crossbred boars (n = 7) were semen collected once per week for four consecutive weeks. Every other collection, the boar's ejaculate was collected into a collection cup and plastic collection bag warmed to 38.5°C containing either no semen extender (control) or 100 mLs of a commercially available long-term semen extender warmed to 38.5°C (treatment). Following collection and processing, the semen was extended to 37.5 × 106 sperm/mL and stored for 6 days post-collection in a semen cooler at 17°C. Motility and morphology were evaluated on day 0 (day of collection) and day 6. There was no day x treatment effect (P > 0.05). Statistical differences (P = 0.03) were found between the treatment and control for sperm motility (82.2 vs. 75.2%) and sperm progressive motility (64.1 vs. 53.5%). No differences (P = 0.96) were present for normal sperm morphology in the treatment compared to the control (89.1 vs. 89.0%). These data suggest that boar semen ejaculates collected into a collection cup and plastic collection bag containing 100 mLs of semen extender warmed to 38.5°C will have greater percentages of motile and progressively motile sperm compared to boar sperm collected into a collection cup and plastic collection bag warmed to 38.5°C containing no semen extender.

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Ameliorative effect of Allium atroviolaceum on sperm quality in cyclophosphamide-treated mice

Future Journal of Pharmaceutical Sciences ◽

10.1186/s43094-021-00234-2 ◽

2021 ◽

Vol 7 (1) ◽

Author(s):

Alireza Hosseini ◽

Mehrdad Shahrani ◽

Shirin Asgharian ◽

Maryam Anjomshoa ◽

Ayoob Rostamzadeh ◽

...

Keyword(s):

Side Effects ◽

Alkylating Agent ◽

Sperm Quality ◽

Sperm Count ◽

Reproductive Toxicity ◽

Herbal Drugs ◽

Sperm Viability ◽

Complementary Treatment ◽

Ameliorative Effect ◽

Alternative Drugs

Abstract Background Cyclophosphamide (CP) is an anti-neoplastic alkylating agent that is extensively used in different chemotherapy regimens. Adverse effects on the reproductive system, especially spermatogenesis, are one of the most important side effects of this drug. It is medically essential to use complementary and alternative drugs. Herbal drugs have long been used as a complementary treatment. Our purpose was to study the effect of hydroalcoholic Allium atroviolaceum L. extract on spermatogenesis in CP-treated mice. Results CP affected a significant decrease in sperm count, motility, viability, and morphology. Sperm count was significantly higher in the all extract groups than in the group of control (p<0.001) and CP group (p<0.001, p<0.01). Sperm motility was significantly greater in the extract (100 and 200mg/kg) groups than in the group of control (p<0.05 and <0.001). Sperm immotility and rotational movement were significantly higher in the CP group than in the CP+extract groups (p<0.001). The sperm viability was significantly greater in the CP+extract (200mg/kg) group than in the CP group (p<0.001). The number of headless sperm, sperm with initial tail, with coiled tail, and sperm with curved body, was significantly lower in the CP+extract (200mg/kg) group than in the CP group (p<0.001). Conclusion A. atroviolaceum extract treatment significantly improved CP-induced reproductive toxicity.

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Viability of Najawa carp (Cyprinus carpio L.) sperm at 4°C temperature storage

E3S Web of Conferences ◽

10.1051/e3sconf/202014701015 ◽

2020 ◽

Vol 147 ◽

pp. 01015

Author(s):

Dimas Fendy Pradana ◽

Ignatius Hardaningsih ◽

Dini Wahyu Kartika Sari

Keyword(s):

Low Temperature ◽

Cyprinus Carpio ◽

Sperm Viability ◽

Low Temperature Storage ◽

Progressive Motility ◽

C Storage ◽

Randomized Design ◽

Significant Difference ◽

Carp Cyprinus Carpio ◽

Temperature Storage

The objectives of this study were to evaluate the sperm viability of Najawa carp (Cyprinus carpio L.) in cryopreservation pre-conditions at 4°C. The design used in this study was Complete Randomized Design with 4 treatments, BSS as a control, 10% DMSO, 0,2 M Sucrose, and 5% DMSO + 0,1 M Sucrose; each consist of three replications. The parameters observed were progressive motility of fresh sperm, diluted sperm before low temperature storage, and 2 hours; 3 hours; 4 hours; 5 hours; one day; one week; a month after 4°C storage. The data were analyzed by ANOVA. The data showed that there was no significant difference between treatment (P>0.05). The best viability was 40.56% of sperm motility which survive for one week, it was achieved by 5% DMSO + 0,1 M Sucrose.

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