scholarly journals Pentoxifylline added to freezing or post-thaw extenders does not improve the survival or in vitro fertilising capacity of boar spermatozoa

Reproduction ◽  
2010 ◽  
Vol 139 (3) ◽  
pp. 557-564 ◽  
Author(s):  
María A Gil ◽  
Marta Hernandez ◽  
Jordi Roca ◽  
Carmen Almiñana ◽  
Xiomara Lucas ◽  
...  
Keyword(s):  
Untreated Control ◽  
Deleterious Effect ◽  
Penetration Rate ◽  
Sperm Quality ◽  
Quality Parameters ◽  
Final Concentration ◽  
Freezing And Thawing ◽  
Boar Spermatozoa

This study evaluated whether pentoxifylline added to freezing and thawing extenders influenced the function of boar spermatozoa. In Experiment 1, pooled ejaculated sperm-rich fractions were frozen in 0.5 ml straws after dilution in extender supplemented with pentoxifylline to a final concentration of 0, 2, 4, 8, 16 or 32 mM. The addition of 4, 8, 16 and 32 mM pentoxifylline to the freezing extender significantly decreased the progressive and total motility of spermatozoa. The percentage of viable spermatozoa with intact acrosomes as well as the penetration rate and the efficiency of fertilisation were significantly lower in pentoxifylline-treated groups compared with the untreated control. In Experiment 2, a pool of three straws with ‘good’ post-thaw sperm quality parameters and another three straws with ‘poor’ sperm quality were diluted in extender with 0, 1, 2, 4, 8, 16 or 32 mM pentoxifylline. Post-thaw samples with both ‘good’ and ‘poor’ sperm quality with 0, 2, 4, 8 and 16 mM were used to assess IVF parameters. The addition of pentoxifylline to post-thaw extender did not improve the post-thaw motility or viability of spermatozoa compared with the control. The in vitro penetration was higher (P<0.05) than the control for oocytes fertilised with spermatozoa that were thawed and incubated in extender with 4, 8 and 16 mM pentoxifylline. However, no differences were observed in the efficiency of fertilisation. We conclude that pentoxifylline, as a supplement added to the freezing extender, has a deleterious effect and that it does not improve the survival or in vitro fertilising efficiency of frozen–thawed boar spermatozoa when added after thawing.

Chlortetracycline fluorescence patterns and in vitro fertilisation of frozen-thawed boar spermatozoa incubated under various bicarbonate concentrations

Zygote ◽  
1997 ◽  
Vol 5 (2) ◽  
pp. 117-125 ◽  
Author(s):  
Lalantha R. Abeydeera ◽  
Hiroaki Funahashi ◽  
Nam-Hyung Kim ◽  
Billy N. Day
Keyword(s):  
Penetration Rate ◽  
Calf Serum ◽  
Cumulus Cells ◽  
In Vitro Fertilisation ◽  
Dependent Manner ◽  
North Carolina State University ◽  
Porcine Oocyte ◽  
Low Incidence ◽  
Boar Spermatozoa

SummaryPorcine oocyte-cumulus complexes were cultured in bovine serum albumin(BSA)-free North Carolina State University (NCSU)23 medium containing porcine follicular fluid (10%), cysteine (0.1 mg/ml)and hormonal supplements (eCG and hCG: 10IU/ml each) for 22h. They were then cultured in the same medium but without hormonal supplements for an additional 22h. After culture, cumulus cells were removed and oocytes were co-incubated with frozen-thawed ejaculated boar spermatozoa in tissue culture medium (TCM)199 containing caffeine (5mM), fetal calf serum (FCS; 10%) and varying concentrations (26–56 mM)of NaHCO3 for 9h(experiment 1). In experiment 2, chlortetracycline (CTC) was used to assess the functional state of spermatozoa incubated under different NaHCO3 concentrations. Experiment 3 examined the effect of FCS (1% and 10%)and NaHCO3 (26 and 46 mM) on fertilisation parameters. Compared with 26 mM, penetration rate was significantly higher (p<0.05)at 36–56mM NaHCO3. Polyspermy showed a similar pattern although no difference was observed between 26 and 36 mM. At 46mM NaHCO3, the mean number of spermatozoa (MNS) penetrated per oocyte increased significantly (p< 0.05). A significantly higher proportion of spermatozoa were capacitated and acrosome reacted at 46 and 56mM NaHCO3, respectively. The fertilisation medium containing 46mM NaHCO3 and 1% FCS showed a higher penetration rate (84%)with a relatively low incidence of polyspermy (39%). The results indicate that NaHCO3 stimulates capacitation and/or the acrosome reaction of boar spermatozoa in a dose-dependent manner and thus affects fertilisation parameters.

scholarly journals Inhibitors of zinc-dependent metalloproteases hinder sperm passage through the cumulus oophorus during porcine fertilization in vitro

Reproduction ◽  
2012 ◽  
Vol 144 (6) ◽  
pp. 687-697 ◽  
Author(s):  
J Beek ◽  
H Nauwynck ◽  
D Maes ◽  
A Van Soom
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Inhibitory Effect ◽  
Fertilization Rate ◽  
Quality Parameters ◽  
Fertilization In Vitro ◽  
Cumulus Oophorus ◽  
Sperm Penetration

In this study, we report for the first time on a possible contribution of metalloproteases in sperm passage through the cumulus matrix in pigs. The presence of 20 μM 1,10-phenanthroline (1,10-PHEN), inhibitor of zinc-dependent metalloproteases, strongly inhibited the degree of sperm penetration in cumulus-intact (CI), but not in cumulus-free (CF), porcine oocytes during IVF. The inhibitory effect of 1,10-PHEN was due to the chelation of metal ions as a non-chelating analog (1,7-PHEN) did not affect IVF rates. Furthermore, incubation with 1,10-PHEN did not affect sperm binding to the zona pellucida nor sperm motility, membrane integrity, or acrosomal status. These findings led to the assumption that 1,10-PHEN interacts with a sperm- or cumulus-derived metalloprotease. Metalloproteases are key players in physiological processes involving degradation or remodeling of extracellular matrix. In vivo, their proteolytic activity is regulated by tissue inhibitors of metalloproteases (TIMP1–TIMP4). We tested the effect of TIMP3 on fertilization parameters after porcine IVF. Similar to 1,10-PHEN, TIMP3 inhibited total fertilization rate of CI but not CF oocytes and did not influence sperm quality parameters. Although the inhibitory effect was stronger in CI oocytes, TIMP3 also reduced the degree of sperm penetration in CF oocytes, suggesting the involvement of a metalloprotease in a subsequent step during fertilization. In conclusion, our results indicate the involvement of TIMP3-sensitive, zinc-dependent metalloprotease activity in sperm passage through the cumulus oophorus in pigs. The results should provide the basis for further biochemical research toward the localization and identification of the metalloprotease involved.

scholarly journals Effect of whole and clarified egg yolk based extenders on post-thaw semen quality in bulls.

2021 ◽  
Author(s):  
Prosper Kamusasa ◽  
Eddington Gororo ◽  
Fungayi Primrose Chatiza
Keyword(s):  
Semen Quality ◽  
Sperm Quality ◽  
Egg Yolk ◽  
Quality Parameters ◽  
Normal Morphology ◽  
Semen Cryopreservation ◽  
Bull Semen ◽  
Inclusion Level ◽  
Whole Egg

Abstract This study was conducted to evaluate the comparative cryoprotective effects of whole egg yolk and clarified egg yolk on post thaw sperm quality parameters and to determine the optimum clarified egg yolk inclusion level (10-20%) in semen extenders for Mashona bull semen cryopreservation. It was shown that there was a significant decrease in sperm quality variables following cryopreservation. Semen quality increased with the concentration of clarified egg yolk, indicating a positive relationship between egg yolk LDL concentration and maintenance of in vitro sperm quality. The 20% clarified egg yolk (CEY20) extender treatment gave post-thaw motility, viability and normal morphology values which were comparable to the control (20% whole egg yolk, WEY20). The 10% clarified egg yolk concentration gave the least post-thaw quality values and the greatest proportion of defective spermatozoa. This experiment found no advantage of replacing whole egg yolk with up to 15% clarified egg yolk in Mashona bull semen cryopreservation. However, 20% clarified and 20% whole egg yolk performed similarly in the maintenance of post-thaw sperm motility, viability and normal morphology.

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

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.

In vitro phagocytosis of boar spermatozoa by neutrophils from peripheral blood of sows

Reproduction ◽  
2000 ◽  
pp. 265-273 ◽  
Author(s):  
A Matthijs ◽  
W Harkema ◽  
B Engel ◽  
H Woelders
Keyword(s):  
Cold Shock ◽  
Female Genital Tract ◽  
Polymorphonuclear Leucocytes ◽  
Freezing And Thawing ◽  
Phagocytosis Assay ◽  
Acrosomal Exocytosis ◽  
Boar Spermatozoa ◽  
Female Genital ◽  
Serum Components

A considerable number of spermatozoa are used in each sow in routine artificial insemination. However, within a few hours after insemination, many spermatozoa are phagocytosed by polymorphonuclear leucocytes. Some aspects of sperm transport in the female genital tract in the sow have been thoroughly investigated, whereas little is known about the mechanisms involved in the phagocytosis of spermatozoa, or about which spermatozoa (fresh, capacitated or dead) are the most susceptible to ingestion by polymorphonuclear leucocytes. In this study, phagocytosis was investigated by use of an in vitro phagocytosis assay. Polymorphonuclear leucocytes were challenged with either untreated, cold-shocked or frozen-thawed spermatozoa, or with spermatozoa that had been treated to induce capacitation in vitro. The influence of serum on phagocytosis was also investigated. Treatment of the semen to induce capacitation in vitro considerably reduced the phagocytosis of spermatozoa, whereas crude treatments like cold-shock or freezing and thawing reduced phagocytosis only in the first 15-30 min of incubation with polymorphonuclear leucocytes. Viable spermatozoa were phagocytosed mainly through a pathway that was independent of complement or other serum components (for example, antibodies). Complement had little effect on phagocytosis of spermatozoa, but did cause acrosomal exocytosis and cell death.

Incubation of boar spermatozoa in viscous media by addition of methylcellulose improves sperm quality and penetration rates during in vitro fertilization

2017 ◽  
Vol 92 ◽  
pp. 14-23 ◽  
Author(s):  
David González-Abreu ◽  
Soledad García-Martínez ◽  
Vanesa Fernández-Espín ◽  
Raquel Romar ◽  
Joaquín Gadea
Keyword(s):  
In Vitro Fertilization ◽  
Sperm Quality ◽  
Vitro Fertilization ◽  
Viscous Media ◽  
Boar Spermatozoa

scholarly journals The Re-Addition of Seminal Plasma after Thawing Does Not Improve Buck Sperm Quality Parameters

Animals ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 3452
Author(s):  
Uchechi Linda Ohaneje ◽  
Uchebuchi Ike Osuagwuh ◽  
Manuel Alvarez-Rodríguez ◽  
Iván Yánez-Ortiz ◽  
Abigail Tabarez ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Quality Parameters ◽  
Mitochondrial Activity ◽  
Dna Integrity ◽  
Vitro Fertilization

In order to achieve a higher post-thaw buck sperm quality, an approach in the thawing protocol of cryopreserved sperm doses under in vitro capacitation conditions mimicking the in vivo female environment was studied. Therefore, functional and kinetic characteristics of buck thawed sperm from males of different ages, the season of collection, and melatonin implanted males in the non-breeding season were assessed after 3 h of incubation in an in vitro fertilization (IVF) media with 20% of buck seminal plasma (SP). Previously, fresh ejaculates were collected via artificial vagina from eight males of the Cabra Blanca de Rasquera breed during two consecutive years in breeding and non-breeding periods. Prior to semen collection in non-breeding seasons, males were split into two groups: one group was implanted with melatonin, while the other was not. In each group, semen samples were pooled, centrifuged, and diluted in an extender containing 15% powdered egg yolk and 5% glycerol before freezing. After thawing, sperm were washed and incubated in three different media: (a) control media (modified phosphate-buffered saline (PBS), (b) IVF commercial media, and (c) IVF media + 20% SP. Sperm motility was evaluated by CASA, while plasma and acrosome membrane integrity, mitochondria activity, and DNA fragmentation were analysed by flow cytometer at 0 h and after 3 h incubation. A significant reduction in motility, mitochondrial activity, plasma, and acrosome membrane integrity were observed after incubation in the presence of SP, although similar to that observed in IVF media alone. DNA integrity was not affected under in vitro capacitation conditions, regardless of SP addition. In conclusion, the addition of SP failed to improve post-thaw buck sperm quality under in vitro conditions irrespective of male age, the season of collection, and melatonin implant.

88 SELECTION OF A CRYOSURVIVAL SPERM POPULATION DOES NOT IMPROVE THE IN VITRO PENETRATING ABILITY OF FROZEN - THAWED BOAR SPERMATOZOA

2008 ◽  
Vol 20 (1) ◽  
pp. 124
Author(s):  
M. L. Perals ◽  
M. A. Gil ◽  
E. M. Garcia ◽  
J. Sanchez-Osorio ◽  
J. M. Vázquez ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Molecular Probes ◽  
Computer Assisted ◽  
Immature Oocytes ◽  
Sperm Analysis ◽  
Boar Spermatozoa ◽  
Casa System ◽  
Selection Of

Boars can be classified as good or bad sperm freezers according to their sperm cryosurvival. Different sperm selection techniques, such as PureSperm� (PS; MidAtlantic Diagnostics, Inc., Mount Laurel, NJ, USA), have been developed to improve functional competence of spermatozoa. The aim of this experimental study was to assess the ability of PS for improving the in vitro penetrating ability of frozen–thawed boar spermatozoa from good and bad sperm freezers. The sperm-rich fractions from two boars, good (Boar A) and bad (Boar B) freezers, were extended in a lactose/eggyolk/ glycerol/Equex Stem (Noba Chemical Sales, Inc., Scituate, ME, USA) mixture (1 � 109 sperm mL–1), dispensed into 0.5-mL straws, and frozen using a programmable cell freezer. After thawing (1.200�C min–1), semen from each boar was split into two aliquots of 500 µL. One aliquot was used as the control. The second was placed into a tube of PS gradient (90%/45%) and centrifuged at 425g for 20 min; the pellet re-suspended in 1 mL of BTS and re-centrifuged at 320g for 10 min (PS sample). Control and PS samples were diluted in supplemented TCM-199 (TCMm; Roca et al. 1998 Reprod. Fertil. Dev. 10, 479–485) at 200 � 106 sperm mL–1. Sperm survival (SV) was assessed afterTCMm dilution according to progressive sperm motility (PSM, %) using a computer-assisted sperm analysis (CASA) system (ISAS�), and plasma and acrosome membrane integrity (PMI; %) by flow cytometry (SYBR�-14/PE-PNA/PI; Molecular Probes, Leiden, The Netherlands). A homologous in vitro penetration (hIVP) assay, using immature oocytes (20 oocytes/2 mL TCMm supplemented with caffeine and calcium lactate), was used to assess sperm penetrating ability (Martinez et al. 1993 Theriogenology 40, 547–557). A total of 960 immature oocytes were inseminated (200 � 103 sperm/oocyte) in 3 batches. After 18 h of co-incubation at 39�C under 5% CO2 in air, the oocytes were washed, mounted on slides, fixed with ethanol:acetic acid (3:1, v/v) for 48 h, stained with 1% lacmoid, and examined under a phase contrast microscope (�400). Oocytes with swollen or unswollen heads of sperm found in the vitellus were considered as penetrated. Sperm penetrability ability (SPA) was assessed according to penetration rate (PR) and the mean number of sperm per oocyte (S/O). Data were analyzed using a PROMIXED model and expressed as mean � SEM. Boar A showed better (P ≤ 0.01) results for both SV and SPA parameters than boar B, independent of sperm treatment. PureSperm improved (P ≤ 0.05) PSM and PMI in both boar A (control v. PS: 48.0 � 5.8 v. 66.5 � 3.6 and 63.1 � 7.7 v. 88.4 � 1.3, respectively) and boar B (12.3 � 1.2 v. 22.2 � 3.7 and 44.3 � 3.5 v. 58.7 � 7.0, respectively). However, no differences (P ≥ 0.05) were observed in PR and S/O in either boar A (71.2 � 3.4 v. 78.3 � 3.1 and 5.0 � 0.4 v. 5.2 � 0.4, respectively) or boar B (34.3 � 3.6 v. 37.3 � 3.9 and 1.5 � 0.1 v. 1.5 � 0.1, respectively). In conclusion, under our laboratory conditions, PureSperm selection improves sperm quality but not in vitro penetrating ability of frozen–thawed spermatozoa of both good and bad sperm freezers. This work was supported by CICYT (AGF2005-00760), Madrid, Spain.

Assessment of sperm quality parameters of six bulls showing different abilities to promote embryo development in vitro

2006 ◽  
Vol 18 (3) ◽  
pp. 395 ◽  
Author(s):  
M. Alomar ◽  
J. Mahieu ◽  
B. Verhaeghe ◽  
L. Defoin ◽  
I. Donnay
Keyword(s):  
Sperm Quality ◽  
Sharp Decrease ◽  
Quality Parameters ◽  
In Vitro Fertilisation ◽  
Sperm Viability ◽  
Fertilisation Rate ◽  
Significant Difference ◽  
Before And After ◽  
Development In Vitro

Experiments were conducted to investigate the possible origins of variation between six bulls showing various blastocyst rates after in vitro fertilisation. No significant difference was observed for the rates of cleavage and 5–8 cell stages, whereas blastocyst yields at Day 6, 7 and 8 post insemination were significantly different between bulls (P < 0.05). Fertilisation rates ranged from 59.5 to 79.3% (P < 0.05), with no difference in the incidence of polyspermy. The proportions of motile and progressive spermatozoa before and after Percoll separation were analysed. A positive effect of Percoll was noted on both parameters (P < 0.05), leading to the absence of difference between bulls after the separation process. Sperm viability and spontaneous acrosome reaction were assessed during 18 h incubation in fertilisation medium. A sharp decrease in sperm viability was observed for all bulls after 2 h incubation, with only 12.6–21.7% of spermatozoa still viable at 18 h. In contrast, the proportion of reacted acrosomes was low in five out of six bulls (<15% at 18 h). In conclusion, the fertilisation rate was the only parameter to show some correlation with blastocyst rate for all bulls.

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