26 SPERM REPRODUCTIVE POTENTIAL OF FETAL FIBROBLAST CELLS ON A NUCLEAR TRANSFER - CLONED BULL AND ON AN ORIGINAL PATERNAL BULL: A COMPARATIVE ANALYSIS

2007 ◽  
Vol 19 (1) ◽  
pp. 131
Author(s):  
S.-H. Bae ◽  
D.-H. Kim ◽  
S.-W. Kim ◽  
G.-S. Im ◽  
M.-J. Lee ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Treated Group ◽  
Flow Cytometer ◽  
Mitochondrial Activity ◽  
Computer Assisted ◽  
Fibroblast Cells ◽  
Sperm Viability ◽  
Fetal Fibroblast ◽  
Reproductive Ability ◽  
Bull Sperm

Recently, cloning technology has produced newborn calves from fetal fibroblast cells and somatic cells. These cloned calves appear to be normal and healthy in terms of reproductive ability. Therefore, we undertook this study to compare the reproductive ability of cloned sperm from the clones themselves and sperm obtained from the frozen semen of the paternal bull (KPN-178). Two cloned bulls from fetal fibroblast cell nuclear transfers (Korean HanWoo Clone-38, 34 months old, and Clone-39, 30 months old) were selected. Sperm from the cloned groups (C-38 and C-39), cloned-38 fresh sperm, and sperm from the paternal bull (KPN-178) were analyzed for the following sperm reproductive characteristics: sperm motility, by computer-assisted sperm analysis (CASA); sperm mitochondrial activity, by flow cytometer after mitotracker staining; and sperm viability, by flow cytometer after staining with live/dead sperm viability kits, both PI and SYBR. Sperm motility was evaluated with Percoll-treated or non-Percoll-treated frozen-thawed spermatozoa. In the Percoll-treated group, total motility of the paternal bull sperm (KPN-178: 75.14%) was significantly lower than that of cloned sperm (C-38: 90.42%; C-39: 87.62%: C-38 fresh: 93.97%; P < 0.05). The non-Percoll-treated group showed different results compared to the Percoll results (P < 0.05), i.e. the total motility of C-38 (92.47%) and C-38 fresh (92.47%) sperm was significantly higher than that of C-39 (79.52%) and KPN-178 (78.41%) sperm. The mitochondrial activity staining rate of KPN-178 (58.22%) sperm was low compared to that of C-38 (88.65%), C-39 (89.45%), C-38 fresh (88.66%) sperm. The sperm viability tests showed similar results (P < 0.05) for all sperm groups: C-38 (PI: 42.53% and SYBR: 46.28%), C-39 (PI: 39.03% and SYBR: 66.18%), C-38 fresh (PI: 34.32% and SYBR: 57.10%), and KPN-178 (PI: 14.43% and SYBR: 47.86%). However, the PI staining of KPN-178 was lower than that of the other sperm groups. Next, in vitro fertilization (IVF) was investigated to compare the reproductive ability of C-38, C-39, KPN-178, and Control (normal bull) sperm. The results, development to the 2-cell stage (157/228, 68.90%; 85/116, 73.28%; 78/114, 68.42%; 61/79, 77.21%, respectively) and blastocyst rates (41/228, 18.0%; 35/116, 30.17%; 26/114, 22.81%; 20/79, 25.32%, respectively) were evaluated (P < 0.05). The IVF rates were not different among cloned bull, paternal bull, and normal bull sperm. A cloned female bovine was successfully impregnated by cloned bull sperm (C-38) using artificial insemination. The calf resulting from this prenancy showed no signs of phenotypical abnormalities. These results suggest that the physiology of cloned bulls and the quality of semen from cloned bulls show no evidence of any deleterious effects.

Lycopene and resveratrol improve post-thaw bull sperm parameters: sperm motility, mitochondrial activity and DNA integrity

Andrologia ◽  
2014 ◽  
Vol 47 (5) ◽  
pp. 545-552 ◽  
Author(s):  
M. N. Bucak ◽  
M. B. Ataman ◽  
N. Başpınar ◽  
O. Uysal ◽  
M. Taşpınar ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Sperm Parameters ◽  
Mitochondrial Activity ◽  
Dna Integrity ◽  
Bull Sperm

scholarly journals Positive effects of trehalose and cysteine on ram sperm parameters

2017 ◽  
Vol 62 (No. 5) ◽  
pp. 245-252 ◽  
Author(s):  
S. Gungor ◽  
C. Ozturk ◽  
AD Omur
Keyword(s):  
Sperm Motility ◽  
Sperm Parameters ◽  
Control Group ◽  
Mitochondrial Activity ◽  
Sperm Viability ◽  
Significant Protection ◽  
Positive Effects ◽  
Liquid Storage ◽  
Acrosome Integrity ◽  
Ram Sperm

The aim of this study was to determine the effects of trehalose and cysteine on sperm motility, viability, mitochondrial activity and acrosome integrity during liquid storage of Merino ram semen. Ejaculates were collected using artificial vaginas from five Merino rams, microscopically evaluated and pooled at 37 °C. The pooled semen samples were diluted in a Tris-based extender, including cysteine (2 mM and 4 mM), trehalose (10 mM and 25 mM) and no antioxidant (control). Diluted semen samples were kept in tubes and cooled from 37 to 5 °C in a cold cabinet, and maintained at 5 °C. Cooled samples were evaluated for sperm motility, viability, mitochondrial activity and acrosome integrity at 0, 24, 48, 72 and 96 h. Extender supplemented with trehalose (10 and 25 mM) and cysteine (2 and 4 mM) led to higher motility in comparison to the control at 24, 48, 72 and 96 h of liquid storage (P < 0.05). Trehalose at the doses of 10 mM, 25 mM and 2 mM cysteine led to higher viability between 24–48–72 h and at 96 h of liquid storage (P < 0.05). Further, 4 mM of cysteine improved sperm viability rates at 24 and 48 h of storage compared to the control group (P < 0.05), and resulted in improved acrosome integrity rates compared to the control group at 72 and 96 h of storage (P < 0.05). Extender supplemented with 10 and 25 mM trehalose at 24 and 72 h and 4 mM cysteine at 24 and 96 h of storage led to higher sperm mitochondrial activity rates when compared to the control group (P < 0.05). In conclusion, the findings of this study show that trehalose and cysteine provided significant protection to ram sperm parameters during liquid storage.

Effects of Red Ginseng Extract on the Epididymal Sperm Motility of Mice Exposed to Ethanol

2011 ◽  
Vol 30 (4) ◽  
pp. 435-442 ◽  
Author(s):  
Mi Jang ◽  
Jin-Woo Min ◽  
Jun-Gyo In ◽  
Deok-Chun Yang
Keyword(s):  
Sperm Motility ◽  
Reproductive Toxicity ◽  
Serum Testosterone ◽  
Treated Group ◽  
Control Group ◽  
Computer Assisted ◽  
Protective Effects ◽  
Red Ginseng ◽  
Ginseng Extract ◽  
Sperm Analysis

The protective effects of red ginseng extract and ginseng wine against ethanol-induced male reproductive toxicity were evaluated in male mice using computer-assisted sperm analysis. Mice were divided into 4 groups of 10 and fed plain saline, 6 g/kg per d of ethanol in saline, red ginseng extract plus ethanol, or a fermented preparation of red ginseng extract daily for 5 weeks. We found that the average seminal vesicle weight was significantly lower in the ethanol-treated group compared to the control group, while those of the ginseng-treated groups tended to be higher than the ethanol-treated group. We found a significant decrease in sperm motility and progressiveness in mice treated with ethanol for 5 weeks, while administration of ethanol plus red ginseng extract appeared to minimize the negative effects of ethanol toxicity on male fertility. Serum testosterone, luteinizing hormone (LH), and follicle stimulating hormone (FSH) were insignificantly lower in the ethanol-treated group than in the control group.

scholarly journals Red-Light Irradiation of Horse Spermatozoa Increases Mitochondrial Activity and Motility through Changes in the Motile Sperm Subpopulation Structure

Biology ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 254 ◽  
Author(s):  
Jaime Catalán ◽  
Marion Papas ◽  
Sabrina Gacem ◽  
Yentel Mateo-Otero ◽  
Joan E. Rodríguez-Gil ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Consumption Rate ◽  
Red Light ◽  
Light Irradiation ◽  
Mitochondrial Activity ◽  
Dna Integrity ◽  
O2 Consumption ◽  
Sperm Viability ◽  
Intracellular Atp ◽  
Atp Levels

Previous studies in other mammalian species have shown that stimulation of semen with red-light increases sperm motility, mitochondrial activity, and fertilizing capacity. This study sought to determine whether red-light stimulation using a light emitting diode (LED) at 620–630 nm affects sperm motility and structure of motile subpopulations, sperm viability, mitochondrial activity, intracellular ATP levels, rate of O2 consumption and DNA integrity of horse spermatozoa. For this purpose, nine ejaculates were collected from nine different adult stallions. Upon collection, semen was diluted in Kenney extender, analyzed, its concentration was adjusted, and finally it was stimulated with red-light. In all cases, semen was packaged in 0.5-mL transparent straws, which were randomly divided into controls and 19 light-stimulation treatments; 6 consisted of a single exposure to red-light, and the other 13 involved irradiation with intervals of irradiation and darkness (light-dark-light). After irradiation, sperm motility was assessed using a Computerized Semen Analysis System (CASA). Flow cytometry was used to evaluate sperm viability, mitochondrial membrane potential and DNA fragmentation. Intracellular levels of ATP and O2 consumption rate were also determined. Specific red-light patterns were found to modify kinetics parameters (patterns: 4, 2-2-2, 3-3-3, 4-4-4, 5-1-5, and 5-5-5 min), the structure of motile sperm subpopulations (patterns: 2, 2-2-2, 3-3-3, and 4-1-4 min), mitochondrial membrane potential (patterns: 4, 3-3-3, 4-4-4, 5-1-5, 5-5-5, 15-5-15, and 15-15-15 min), intracellular ATP levels and the rate of O2 consumption (pattern: 4 min), without affecting sperm viability or DNA integrity. Since the increase in some kinematic parameters was concomitant with that of mitochondrial activity, intracellular ATP levels and O2 consumption rate, we suggest that the positive effect of light-irradiation on sperm motility is related to its impact upon mitochondrial activity. In conclusion, this study shows that red LED light stimulates motility and mitochondrial activity of horse sperm. Additional research is needed to address the impact of red-light irradiation on fertilizing ability and the mechanisms through which light exerts its effects.

scholarly journals 448 Late-breaking: GWAS analysis show QTL in horses which are characterized by sperm resistance to freezing

2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 119-120
Author(s):  
Andrei A Kudinov ◽  
Natalia Dementieva ◽  
Elena Nikitkina ◽  
Michail Atroshchenko ◽  
Artem Musidray
Keyword(s):  
Sperm Motility ◽  
Association Studies ◽  
Semen Analysis ◽  
Individual Variability ◽  
Mitochondrial Activity ◽  
Computer Assisted ◽  
Russian Science ◽  
Progressive Motility ◽  
Genome Wide ◽  
Selection Of

Abstract Damaging effects of low temperatures on the functionality of sperm is well known. Stallion sperm cryoresistance has a high individual variability. The genetic basis of cryoresistance and its heritability are little studied. The aim of the study was to search for genome-wide associations with sperm motility after freezing. 96 sperm samples were collected. The collected sperm was diluted to a final concentration 100 mln/ml and frozen in 0.5 ml straws or 18–20 ml tubes. The evaluation of semen was carried out no earlier than 24 hours after freezing. Sperm motility was assessed by computer-assisted semen analysis (CASA). Genomic DNA was purified form semen samples and genotyped using Affymetrix Equine HD array. Genotypes quality control and association studies were performed in Plink 1.9 and EMMAX software respectively. Evaluation of sperm motility showed high individual variability in both total and progressive motility after freezing. Total motility varied from 0.5 to 92.5%, and progressive motility from 0.5 to 70.8%. Sufficient associations of PTF with SNP’s were detected on chromosome (Chr) 1 (P < 1.33e-09) and 4 (P < 2.00e-09). Found SNP’s were located in genes PAS domain-containing protein 3 (Chr1) and UBAP1-MVB12 (Chr4). Expression of these genes in human body was found in cerebrum and male genitals. Suggestive SNPs were found lying nearby to genes responsible for formation of cell wall and proteins affecting mitochondrial activity. Performed studies are presenting first step of understanding genetic background of cryoresistancy of semen in horses. Found markers could be used for selection of stallions based on cryopreservation ability. Authors acknowledge financial support from Russian Science Foundation, Grant No: 18-16-00071.

scholarly journals Suppementation of Kelor Leaf (Moringa oleifera) Aqueous Extract Increase on Post-Thawed Limousin Bull Sperm Quality

2021 ◽  
Vol 4 (2) ◽  
pp. 249
Author(s):  
Syuhuud Arumbinang Wajdi ◽  
Budi Utomo ◽  
Rimayanti Rimayanti ◽  
Erma Safitri ◽  
Tri Wahyu Suprayogi ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Aqueous Extract ◽  
Moringa Oleifera ◽  
Sperm Quality ◽  
Skim Milk ◽  
Egg Yolk ◽  
Sperm Viability ◽  
Bull Sperm

The purpose of this research was to determine the best dosage of Moringa oleifera Aqueous extract in egg yolk skim milk extender for post thawed Limousin Bull sperm quality on aspect viability, and the level of. The treatment was divided into five groups: egg yolk and skim milk diluter (P0), 2,5% M. oleifera aqueous ekstract in 4 ml egg yolk skim milk (P1), 5% M. oleifera aqueous ekstract in 4 ml egg yolk skim milk (P2), 10% M. oleifera aqueous ekstract in 4 ml egg yolk skim milk (P3), 20% M. oleifera aqueous ekstract in 4 ml egg yolk skim milk (P4). The sperm quality was observed post thawing. The data were analyzed using ANOVA and Duncant Test. The best sperm motility showed on P2 with 43b ± 5,70, the best sperm viability showed on P3 with 58,20b ± 8,72 and than the lowest level of malondialdehyde showed on P4 with 5,434a ± 1,034. In conclusion addition of M. oleifera on dose 10% can increase quality of Limousin Sperm Post Thawed.

scholarly journals Effect of refrigeration systems upon frozen bull sperm viability assessed by computer-assisted sperm analysis and fluorescent probes

2012 ◽  
Vol 33 (5) ◽  
pp. 1923-1930
Author(s):  
Marcelo George M Chacur ◽  
Huberson Sanches Dias ◽  
Patricia de Mello Papa ◽  
Frederico Ozanam Papa ◽  
Cely M Melo-Ona
Keyword(s):  
Fluorescent Probes ◽  
Computer Assisted ◽  
Sperm Viability ◽  
Sperm Analysis ◽  
Computer Assisted Sperm Analysis ◽  
Bull Sperm

39 General motility and mitochondrial cytochemical activity of post-thawed semen of pasture-fed Nelore bulls supplemented with palm and soybean oils

2019 ◽  
Vol 31 (1) ◽  
pp. 145
Author(s):  
P. P. Tsuneda ◽  
L. K. Hatamoto-Zervoudakis ◽  
T. F. Motheo ◽  
J. T. Zervoudakis ◽  
M. Nichi
Keyword(s):  
Sperm Motility ◽  
Lateral Displacement ◽  
Young Male ◽  
Mitochondrial Activity ◽  
Computer Assisted ◽  
Sperm Cells ◽  
Activity Class ◽  
Sperm Analysis ◽  
Soybean Oils ◽  
Progressive Motility

This study aimed to evaluate general motility and mitochondrial cytochemical activity of post-thawed semen of pasture-fed Nelore bulls supplemented with palm and soybean oils (protected by calcium salts). Twenty-four young male Nellore bulls were randomly assigned to 2 groups: control (CO; corn supplement and distillery grains without the addition of protected fat) and palm and soybean oils association (OP+OS; control supplement+145g of protected soybean oil+145g of protected palm). The experiment lasted 84 days, and semen was collected for cryopreservation. Post-thawed general sperm motility was assessed by computer-assisted sperm analysis using a computerized analyser (HTR Ivos II; Hamilton Thorne, Beverly, MA, USA). Motility (%), progressive motility (%), velocity average path (μm s−1), linearity (%), and lateral displacement of sperm head (μm) were analysed. Additionally, mitochondrial cytochemical activity was evaluated by co-incubation with 3,3′-diaminobenzidine (DAB; 1mg mL−1 of PBS) at 37°C (water bath) for 1h under reduced lighting conditions. Sperm cells were classified based on the mitochondrial activity of their midpiece in a 4-class scale: class 1 (DAB 1; all mitochondria are active; sperm cells with midpiece completely stained), class 2 (DAB 2; sperm cells with active and inactive segments but with prevalence of active stained segments, indicating average to high mitochondrial activity), class 3 (DAB 3; spermatozoa with less than half of the mitochondrial sheath active), and class 4 (DAB 4; spermatozoa totally inactive, with midpiece completely discolored). Data were analysed using SAS statistical program (SAS 9.3, SAS Institute Inc., Cary, NC, USA), and Tukey’s test was used to identify treatments. Study design was completely randomized, and effects were declared significant at P<0.05. Increased percentage of sperm motility (CO: 30.27±5.44v. OP+OS: 38.90±4.68) and progressive motility (CO: 17.27±3.84v. OP+OS: 22.90±3.88) were noticed in post-thawed samples after supplementation. Nevertheless, the addition of palm and soybean oils to the feed promoted a decrease in the percentage of DAB 3 sperm cells (CO: 4.77±1.03v. OP+OS: 2.06±0.51; P=0.0267). Therefore, supplementation with palm and soybean oils improves sperm kinetics in post-thawed semen of grazing Nellore bulls.

Prevention and elimination of bovine viral diarrhea virus infections in fetal fibroblast cells

2004 ◽  
Vol 64 (2) ◽  
pp. 113-118 ◽  
Author(s):  
M GIVENS ◽  
D STRINGFELLOW ◽  
C DYKSTRA ◽  
K RIDDELL ◽  
P GALIK ◽  
...  
Keyword(s):  
Bovine Viral Diarrhea Virus ◽  
Bovine Viral Diarrhea ◽  
Fibroblast Cells ◽  
Virus Infections ◽  
Fetal Fibroblast ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Viral Diarrhea Virus

scholarly journals Post-Thaw Sperm Quality and Functionality in the Autochthonous Pig Breed Gochu Asturcelta

Animals ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1885
Author(s):  
José Néstor Caamaño ◽  
Carolina Tamargo ◽  
Inmaculada Parrilla ◽  
Felipe Martínez-Pastor ◽  
Lorena Padilla ◽  
...  
Keyword(s):  
Genetic Resource ◽  
Sperm Quality ◽  
Genetic Material ◽  
Mitochondrial Activity ◽  
Computer Assisted ◽  
Kinematic Parameters ◽  
Linear Mixed Effects ◽  
Sperm Analysis ◽  
Computer Assisted Sperm Analysis ◽  
Analysis System

Genetic resource banks (GRB) preserve the genetic material of endangered, valuable individuals or genetically relevant breeds. Semen cryopreservation is a crucial technique to reach these goals. Thus, we aimed to assess the sperm parameters of semen doses from the native pig breed Gochu Asturcelta stored at the GRB of Principado de Asturias (GRB-PA, Gijón, Spain), focusing on intrinsic and extrinsic (boar, season) factors. Two straws per boar (n = 18, 8–71 months of age) were thawed, pooled, and assessed after 30 and 150 min at 37 °C by CASA (computer-assisted sperm analysis system; motility and kinematic parameters) and flow cytometry (viability, acrosomal status, mitochondrial activity, apoptosis, reactive oxygen species, and chromatin status). The effects of age, incubation, and season on post-thawing quality were determined using linear mixed-effects models. Parameters were on the range for commercial boar breeds, with chromatin status (SCSA: fragmentation and immaturity) being excellent. Incubation decreased sperm quality and functionality. The boar age did not have a significant effect (p > 0.05), but the between-boar variability was significant (p < 0.001). The season significantly affected many parameters (motility, kinematics, viability, acrosomal status, mitochondrial activity), especially after 150 min of incubation. In general, samples collected in spring and summer showed higher quality post-thawing, the lowest in winter. In conclusion, the sperm doses from the Gochu Asturcelta breed stored at the GRB-PA showed excellent chromatin status and acceptable characteristics after thawing. Therefore, boar and seasonal variability in this autochthonous breed could be relevant for cryobank management.

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