scholarly journals Effects of insulin on sperm cell quality in ram semen cooled at 5°C

2021 ◽  
Vol 42 (6supl2) ◽  
pp. 3803-3812
Author(s):  
Maurício Fraga van Tilburg ◽  
◽  
Rodrigo Vasconcelos de Oliveira ◽  
Carla Renata Figueiredo Gadelha ◽  
Bruno Fagundes ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Semen Analysis ◽  
Control Group ◽  
Computer Assisted ◽  
Insulin Group ◽  
Hypoosmotic Swelling Test ◽  
Semen Extender ◽  
Ram Sperm ◽  
Acrosomal Integrity

Insulin is present in the seminal plasma and is involved in sperm activities like motility and capacitation. However, the effects of insulin on the viability of cooled ram sperm are not fully understood. Therefore, the objective of the current study was to evaluate the effect of insulin addition on ram sperm maintained at 5ºC. Sperm samples were collected from six healthy, mature Santa Inês rams. The ejaculates were divided into two aliquots with (insulin group) or without (control group) insulin (3 IU mL-1) in the semen extender, and then cooled at 5°C for 48 hours. Subsequently, the sperm cells were evaluated for motility and kinetics using computer-assisted semen analysis. The samples were evaluated for acrosomal integrity by fluorescein using isothiocyanate combined with peanut agglutinin (FITC-PNA) and membrane functionality by the hypoosmotic swelling test. The semen analysis was performed after 24 or 48 hours of cooling. There was an increased percentage of progressive sperm motility (%), straightness (%), linearity (%) and beat caudal frequency (Hz) in the insulin group after 24 and 48 hours of cooling (p < 0.05). However, insulin did not affect total sperm motility, sperm velocities (VSL, VAP and VCL) (μm seg-1), acrosomal integrity and membrane functionality during cooling (p > 0.05). In conclusion, the addition of 3 IU mL-1 insulin to ram semen extender improved the quality of sperm motility after cooling.

84 EVALUATION OF BULL SEMEN AT 1 H VS. 48 H OF POST-FROZEN STABILIZATION TIME

2006 ◽  
Vol 18 (2) ◽  
pp. 150
Author(s):  
G. M. Brogliatti ◽  
G. Larraburu ◽  
R. Cavia ◽  
M. E. Carini
Keyword(s):  
Liquid Nitrogen ◽  
Artificial Insemination ◽  
Semen Analysis ◽  
Computer Assisted ◽  
Stabilization Time ◽  
Bull Semen ◽  
Straight Line ◽  
Semen Extender ◽  
Lateral Head

The process of cryopreservation of bull semen in liquid nitrogen at −196°C is usually carried out after 3 to 6 h of refrigeration at 4°C post-collection. To guarantee the quality of the final product, the frozen straws are evaluated after cryopreservation. The seminal samples are usually stabilized during 48 h before being analyzed (Hafez, Reproduction and Artificial Insemination in Animals, 1989); this would retard the possible commercialization. The objective of the present study was to determine motility parameters and viability of semen doses stabilized by 1 h or more than 48 h in liquid nitrogen at −196°C. A total of 122 ejaculated from 23 different adult bulls (Angus, Brangus, Braford, and Hereford) were evaluated in an artificial insemination center between January and April 2005. The semen was diluted in a semi-defined semen extender (Andromed, Minitub, Germany) and frozen in an automatic freezer (Digicool, IMV, France). Parameters of velocity average path (VAP, μm/s), velocity straight line (VSL, µm/s), amplitude lateral head (ALH, µm), linearity (LIN, %), percentage of rapid cells (RAPID, %), and viability (VIA, %) were determined by Computer Assisted Semen Analysis (CASA, HTM-ceros 12.1, Berkeley, CA, USA). The obtained results were analyzed statistically with T Student and are summarized in Table 1. The results indicate that there is no difference in the velocity of the spermatozoa evaluated 1 h or 48 h post-frozen. There is no difference in VAP, VSL, movement of amplitude lateral head (ALH), or linearity (LIN). The percentage of viable spermatozoa was not affected in either group. Statistical analysis indicates that there is no difference (P > 0.05) in any of the evaluated parameters. The results demonstrate that spermatic motility and viability of frozen bull semen could be evaluated before 48 h post-frozen. This allows reduction of the time between freezing and evaluation and immediate availability of the bull straws. Table 1. Parameters of motility and viability at 1 h vs. 48 h of post-frozen stabilization time This research was supported by Centro Genético Bovino EOLIA S.A.

55 EFFECT OF TREHALOSE FOR FROZEN–THAWED RAM SPERM MOTILITY PARAMETERS

2015 ◽  
Vol 27 (1) ◽  
pp. 120
Author(s):  
M. M. Toishibekov ◽  
M. T. Jazkbayev ◽  
B. B. Molzhigitov
Keyword(s):  
Sperm Motility ◽  
Semen Analysis ◽  
Sperm Concentration ◽  
Egg Yolk ◽  
Computer Assisted ◽  
Progressive Motility ◽  
Indigenous Sheep ◽  
Standard Tool ◽  
Freezing Curve ◽  
Ram Sperm

Computer-assisted sperm analysers have become the standard tool for evaluating sperm motility because they provide objective results for thousands of mammalian spermatozoa. Ram semen was collected using electro-ejaculation from 10 adult rams of Chingizskaya indigenous sheep breed. Motility was determined using computer-automated semen analysis (Hamilton Thorne Motility Analyzer, Beverly, MA, USA). Trehalose solution (0.375 M) was added to Tris-buffered saline solution to give the following trehalose extenders: 25, 50, 75, and 100% (vol:vol), and analysed for motility using computer-automated semen analysis. The sperm pellets were resuspended at 24°C in cooling extender – trehalose extenders of each concentration containing 5% egg yolk. The diluted semen was cooled to 5°C within 2 h. The semen was then further diluted 1 : 1 with freezing extender – each trehalose extender containing 1.5% glycerol to obtain a sperm concentration of 2.0 × 108 cells mL–1 – and then loaded into 0.5-mL straws. Straws were frozen using a programmable freezer with a freezing curve of 5°C to –5°C at 4°C per min, –5°C to –110°C at 25°C per min, and –110°C to –140°C at 35°C per min, and then the straws were plunged into liquid nitrogen for storage. Frozen samples were thawed in a 37°C water bath for 30 s and analysed for motility using computer-automated semen analysis. Statistical analyses were performed with a Student's test. The fresh semen samples showed the next results: motility 88.3 ± 2.4%, progressive motility 26.8 ± 6.9%, and progressive velocity 61.9 ± 4.2 μm s–1. Motility of the frozen-thawed spermatozoa was 63.6 ± 2.9% (25% trehalose), 55.6 ± 5.2% (50%), 32.4 ± 4.7% (75%), and 23.6 ± 3.2 (100%). Progressive motility was 15.6 ± 3.9% (25%), 13.7 ± 3.7% (50%), 4.5 ± 1.3% (75%), and 5.2 ± 1.3% (100%). Progressive velocity was 93.5 ± 8.3 μm s–1 (25%), 85.4 ± 8.1 μm s–1 (50%), 65.7 ± 6.1 μm s–1 (75%), 35.2 ± 3.3 μm s–1 (100%). Motility of the frozen-thawed spermatozoa significantly decreased with increasing concentrations of trehalose in the extender (P < 0.05). These preliminary studies showed that further research is needed of use trehalose for ram spermatozoa cryoconservation.

97 COMPARISON OF MOTILITY AND ACROSOMAL INTEGRITY OF ELECTRO-EJACULATED AND EPIDIDYMAL RAM SPERM AFTER EXPOSURE TO VARIOUS ANISOSMOTIC SOLUTIONS, CRYOPROTECTIVE AGENTS, AND TEMPERATURES

2008 ◽  
Vol 20 (1) ◽  
pp. 129
Author(s):  
O. Varisli ◽  
C. Uguz ◽  
C. Agca ◽  
J. Green ◽  
Y. Agca
Keyword(s):  
Sucrose Solution ◽  
Semen Analysis ◽  
Chilling Stress ◽  
Molecular Probes ◽  
Current Data ◽  
Epifluorescence Microscopy ◽  
Computer Assisted ◽  
Cryoprotective Agents ◽  
Ram Sperm ◽  
Acrosomal Integrity

Effective ram sperm cryopreservation protocols, which would yield acceptable lambing rates following AI, are currently lacking. The objectives of the current studies were to compare the effects of various anisosmotic conditions, cryoprotective agents (CPA), and temperatures on the motility and acrosomal integrity of electro-ejaculated and epididymal ram sperm. Three experiments were conducted. In Experiment 1, electro-ejaculated and epididymal ram sperm were exposed to 75, 150, 225, 600, 900, and 1200 mOsm kg–1 sucrose solutions, held for 5 min, and then returned to isosmotic conditions. Motility characteristics of sperm during exposure to each anisosmotic solution and after return to isosmotic conditions were determined. In Experiment 2, electro-ejaculated and epididymal ram sperm were exposed to 1 m glycerol (Gly), dimethyl sulfoxide (DMSO), ethylene glycol (EG), or propylene glycol (PG) for 5 min and then returned to isosmotic conditions. Motility characteristics of sperm samples during exposure to each CPA solution and after return to isosmotic conditions were determined. In Experiment 3, effects of various temperatures on motility characteristics of electro-ejaculated and epididymal ram sperm were determined after exposure to three different sub-physiologic temperatures (4, 10, and 22�C) for 30 min and subsequently return to 37�C. A computer-assisted semen analysis system was used to determine sperm motility characteristics. Epifluorescence microscopy was used to determine sperm acrosomal integrity after fixing and staining with Alexa Fluor-488-PNA (Molecular Probes, Eugene, OR, USA). The data were analyzed by ANOVA by using SAS (SAS Institute, Inc., Cary, NC, USA). The motility of electro-ejaculated ram sperm was significantly more affected by anisosmotic stress than was that of epididymal ram sperm (P < 0.05). While anisosmotic stress had no effect on acrosomal integrity of epididymal ram sperm, there was a significant reduction in acrosomal integrity for electro-ejaculated ram sperm after the addition and removal of a 75 mOsm sucrose solution. The abrupt addition and removal of 1 m Gly, DMSO, EG, or PG had no effect on the motility and acrosomal integrity of epididymal ram sperm (P > 0.05). However, there was a slight decrease in acrosomal integrity for electro-ejaculated ram sperm after exposure to 1 m Gly, DMSO, or EG (P > 0.05). Both epididymal and electro-ejaculated ram sperm exhibited temperature-dependent loss of motility and acrosomal integrity (P < 0.05). However, electro-ejaculated ram sperm were more sensitive to chilling stress than were epididymal sperm (P < 0.05). In conclusion, current data overall suggest that while epididymal ram sperm are extremely resilient to various cryobiologically relevant stress conditions, ejaculated ram sperm demonstrate greater sensitivity to such stressors. These current findings should be taken into account when handling and developing cryopreservation protocols for ejaculated and epididymal ram sperm.

scholarly journals Effect of quercetin or butylated hydroxytoluene on cooled or frozen-thawed ram sperm quality

2022 ◽  
Vol 43 (2) ◽  
pp. 841-854
Author(s):  
Lucas Emanuel Ferreira Canuto ◽  
◽  
Lorenzo Garrido Teixeira Martini Segabinazzi ◽  
Endrigo Adonis Braga de Araújo ◽  
Luis Fernando Mercês Chaves Silva ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Butylated Hydroxytoluene ◽  
Epifluorescence Microscopy ◽  
Computer Assisted ◽  
Semen Extender ◽  
Ram Sperm ◽  
Motility Parameters

Cooling and freezing processes cause physical and chemical damage to sperm by cold shock and oxidative stress. This study aimed to evaluate the effect of two antioxidants on sperm parameters of cooled and frozen-thawed ram semen diluted in an egg yolk-based extender. Semen was collected from 30 rams and processed in two consecutive experiments to test the inclusion of different concentrations of quercetin and butylated hydroxytoluene (BHT) in an egg yolk-based semen extender. Dimethyl sulfoxide (DMSO) was added as a solvent to the semen extender in a ratio of 1 mL DMSO for 90 mg of quercetin and 1 mL DMSO for 880 mg of BHT. After collection, semen was diluted at 200 × 106 motile sperm/mL (control) and split into different groups in each experiment. In experiment 1, semen was diluted with the extender containing quercetin (Q5, 5 μg/mL; Q10, 10 μg/mL; Q15, 15 μg/mL) or DMSO alone (DMSO1, 0.055 μL DMSO per mL; DMSO2, 0.165 μL DMSO per mL). In experiment 2, semen was diluted with the extender with BHT (BHT1, 0.5 μg/mL; BHT2, 1 μg/mL; BHT3, 1.5 μg/mL) or DMSO alone (DMSO3, 0.375 μL DMSO per mL; DMSO4, 1.125 μL DMSO per mL). After dilution, the semen was divided into two aliquots. Treated ram sperm samples were also subjected to different storage methods. The first set of samples was cooled at 5 °C for 24 h, whereas the second set of samples was frozen-thawed. Sperm motility parameters and plasma membrane integrity (PMI) were evaluated immediately after dilution (0h) and 24 h after cooling and in the frozen-thawed samples via computer-assisted sperm analysis and epifluorescence microscopy, respectively. The inclusion of quercetin or BHT did not affect sperm motility parameters or PMI of fresh, cooled, or frozen-thawed sperm in this study (P < 0.05). However, further studies are needed to test the effects of these antioxidants on the fertility of cryopreserved ram semen.

Effect of separate and combined exposure of selenium and diazinon on rat sperm motility by computer assisted semen analysis

2016 ◽  
Vol 38 ◽  
pp. 144-149 ◽  
Author(s):  
Robert Toman ◽  
Svatoslav Hluchy ◽  
Michal Cabaj ◽  
Peter Massanyi ◽  
Shubhadeep Roychoudhury ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Semen Analysis ◽  
Computer Assisted ◽  
Combined Exposure

Impact of supplementing commercial rabbit pellets with fresh Moringa oleifera leaves on semen characteristics in rabbit bucks under tropical climate in Trinidad

2021 ◽  
Author(s):  
Krishna Mohan Kumar
Keyword(s):  
Sperm Motility ◽  
Moringa Oleifera ◽  
Sperm Count ◽  
Control Group ◽  
Computer Assisted ◽  
Group Iii ◽  
Group I ◽  
Significant Difference ◽  
Moringa Oleifera Leaves ◽  
The Impact

Objective This study aimed to evaluate the impact of the dietary supplement of Moringa oleifera leaves (MOL) on semen quality and characteristics in rabbits. Methods Eighteen (n=18) breeding bucks of New Zealand white, of similar age group, were used for the study. Three feeding regimes, (i) 100% commercial rabbit pellets (CRP)-Group I (ii) 90% CRP + 10% fresh MOL on a dry matter (DM) basis – Group II and (iii) 80% CRP + 20% fresh MOL on a DM basis – Group III, were adopted and the trial continued for 21 days. After adaptation to the diet, semen was collected from each buck and subjected to evaluation using a computer-assisted semen analyser. Results In Group III, the sperm count, normal sperm morphology, and sperm motility increased (52.0%) in comparison with the control (Group I; 50.1%). The inclusion of 20% Moringa oliefera in the diet (Group III) caused a significant increase (P<0.05) in semen concentration (Control =136.2 M/mL; Group III=297.2 M/mL). There was no significant difference (P>0.05) in sperm motility and semen volume among the groups. Conclusion The results suggest that supplementing commercial rabbit pellets with 20% fresh Moringa oliefera leaves on a DM basis can improve the quality and characteristics of semen in breeding bucks.

scholarly journals iTRAQ-Based Sperm Proteomic Analyses Reveals Candidate Proteins Affecting the Quality of Spermatozoa From Boar on Plateaus

2020 ◽  
Author(s):  
Yanling Zhao ◽  
Yaomei Wang ◽  
Feipeng Guo ◽  
Bo Lu ◽  
Jiale Sun ◽  
...  
Keyword(s):  
Western Blot ◽  
Cellular Response ◽  
Sperm Quality ◽  
Semen Analysis ◽  
Estrogen Signaling ◽  
Glutathione Metabolism ◽  
Computer Assisted ◽  
Expression Levels ◽  
Relative Expression

Abstract Background: Tibetan pigs (TP) exhibit heritable adaptations to their hypoxic environments as a result of natural selection. Whereas, what candidate proteins affecting the sperm quality of boar on plateaus has not been clearly investigated yet. Methods: In this study, to reveal the candidate proteins affecting the quality of spermatozoa from boar on plateaus, we analyzed the sperm quality by Computer-assisted semen analysis (CASA) system and Reactive oxygen species (ROS) levels, compared the sperm proteomes between TP and Yorkshire pigs (YP) raised at high altitudes using the isobaric tags for relative and absolute quantitation (iTRAQ) in combination with the liquid chromatography-tandem mass spectrometry (LC-MS/MS) proteomic method, and confirmed the relative expression levels of the four proteins by western blot. Results: The sperm quality of the TP was superior to that of the YP on plateaus. Of 1,555 quantified proteins, 318 differentially expressed proteins (DEPs) were identified. Gene ontology (GO) analysis revealed that the DEPs were predominantly associated with the sorbitol metabolic process, removal of superoxide radicals, cellular response to superoxide, response to superoxide and regulation of the mitotic spindle assembly. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were mainly enriched in pathways involved in the regulation of the actin cytoskeleton, glutathione metabolism, oxidative phosphorylation, and estrogen signaling. And based on the protein-protein interaction (PPI) network analysis, we identified 8 candidate proteins (FN1, EGF, HSP90B1, CFL1, GPX4, NDUFA6, VDAC2, and CP) that might play important roles that affect the sperm quality of boar on plateaus. Moreover, the relative expression levels of the proteins (CFL1, EGF, FN1, and GPX4) were confirmed by western blot. Conclusions: Our results reveal 8 candidate proteins (FN1, EGF, HSP90B1, CFL1, GPX4, NDUFA6, VDAC2, and CP) affecting the sperm quality of boar on plateaus, providing a reference for studies on improving sperm quality and the molecular breeding of boar on plateaus.

scholarly journals Changes in Quality of Native and Frozenthawed Semen in Relation to Two Collections Performed in a 24-hour Interval and Adition of Clarified Egg Yolk to Extender

2016 ◽  
Vol 47 (2) ◽  
pp. 60-67
Author(s):  
P. Folková ◽  
J. Šichtař ◽  
O. Šimoník ◽  
A. Dokoupilová ◽  
R. Rajmon
Keyword(s):  
Sperm Motility ◽  
Egg Yolk ◽  
Semen Collection ◽  
Semen Extender

Abstract The aim of the study was to evaluate the effect of repeated semen collection and the substitution of normal egg yolk with clarified egg yolk to commercially produced semen extender on qualitative parameters of frozen-thawed canine semen. Two semen collections were scheduled in a 24-hour interval and in each of six dogs, three 1st and three 2nd collections were performed. The frozen-thawed sperm samples were prepared either with clarified or normal egg yolk and motility and viability were evaluated. The effect of the sequence of semen collection was demonstrated by significant differences in motility and also in viability of sperms both in native and frozen-thawed ejaculate. The percentage of viable sperms was significantly higher in samples from the 2nd compared to the 1st collection. This trend was the same also in motility except in native ejaculate. The addition of clarified egg yolk was beneficial for higher survival of sperms immediately after thawing and also after 30 min of incubation, compared to samples with normal egg yolk. Sperm motility evaluated after thawing was higher in samples with clarified egg yolk, without an apparent connection with semen collection sequence. The decrease of values of the qualitative parameters of sperms observed in the period of 30 min of incubation was significantly slowed down when clarified egg yolk was used. This was especially obvious in samples from the 2nd collection.

scholarly journals Ram sperm motility after intermittent scrotal insulation evaluated by manual and computer-assisted methods

2006 ◽  
Vol 8 (4) ◽  
pp. 411-418 ◽  
Author(s):  
C. Arman ◽  
P. I. Quintana Casares ◽  
L. G. Sanchez-Partida ◽  
B. P. Setchell
Keyword(s):  
Sperm Motility ◽  
Computer Assisted ◽  
Scrotal Insulation ◽  
Ram Sperm
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