scholarly journals In vitro addition of docosahexaenoic acid improves the quality of cooled but not frozen–thawed stallion semen

2017 ◽  
Vol 29 (10) ◽  
pp. 2021
Author(s):  
D. M. Silva ◽  
S. A. Holden ◽  
A. Lyons ◽  
J. C. Souza ◽  
S. Fair
Keyword(s):  
Lipid Peroxidation ◽  
Vitamin E ◽  
Docosahexaenoic Acid ◽  
Membrane Fluidity ◽  
Progressive Motility ◽  
Frozen Semen ◽  
Acrosome Integrity

The aim of the present study was to assess the effect of the addition of docosahexaenoic acid (DHA) on the in vitro quality of cooled and frozen–thawed stallion semen. In Experiment 1, semen from 10 stallions was collected (three ejaculates per stallion). Semen was diluted to 100 × 106 spermatozoa mL–1 with 0.02 mM vitamin E (VE) and 0, 1, 10 or 20 ng mL–1 DHA and frozen. Semen was thawed and total motility (TM), rapid progressive motility (PM), acrosome integrity, membrane fluidity and morphology were assessed. In Experiment 2, semen from three stallions was collected (three ejaculates per stallion) and frozen as in Experiment 1, but VE and DHA were added after thawing. TM and PM were assessed at 30, 60 and 120 min and viability, acrosome integrity and membrane fluidity were evaluated at 30 min. In Experiment 3, semen from five stallions was collected (one to three ejaculates per stallion), diluted to 20 × 106 spermatozoa mL–1 and stored at 4°C. After 1, 24, 48 and 72 h, TM, PM, viability, membrane fluidity and lipid peroxidation were assessed. The addition of DHA had no effect on frozen semen (Experiments 1 and 2) but improved TM, PM and membrane fluidity in cooled stallion semen.

scholarly journals Effect of Oral Administration of Selenium and Vitamin E on the Quality of Fresh, Refrigerated and Frozen Semen in French Bulldog Breed Dogs

2017 ◽  
Vol 45 (1) ◽  
pp. 7
Author(s):  
Marcelo George Mungai Chacur ◽  
Mariana Grandis Ripari de Souza ◽  
Camila Dutra de Souza ◽  
Camila Pires Cremasco
Keyword(s):  
Vitamin E ◽  
Citric Acid ◽  
Oral Administration ◽  
Liquid Nitrogen ◽  
Sperm Concentration ◽  
Oral Supplementation ◽  
Frozen Semen ◽  
French Bulldog ◽  
Fresh Semen

Background: New methodologies have been developed seeking to maximize pregnancy rate in female dogs created in commercial kennels, and also in order to maintain the quality of canine semen after dilution, refrigeration or freezing. One of the main factors that generate damage to sperm is oxidative stress, to minimize sperm damage, selenium and antioxidants like vitamin E are administered, by oral administration, seeking to improve the quality of semen. The objective was to study the effect of vitamin E and selenium, by oral administration, in the quality of fresh, refrigerated and frozen semen in adult dogs French Bulldog breed.Materials, Methods & Results: Semen samples were collected from 5 adult dogs, French Bulldog breed, being 2 semen drawing before the daily oral supplementation with vitamin E and selenium (ESE®) and semen drawing at 20, 40 and 60 days after the beginning of oral supplement. The ejaculated samples were diluted in TRIS - fructose citric acid (3.28 g TRIS-hydroxy-methyl-amino-methane, 1.78 g of citric acid monohydrate and 1.25 g of D - fructose, dissolved in 100 mL of distilled water and added of 20% egg yolk and 6% of glycerol. The characteristics evaluated in fresh semen were: volume (mL), color, appearance, concentration (x106 / mL), sperm motility (%), sperm strength (1 to 5) and morphology (%). For refrigerated and frozen semen were analyzed: sperm motility (%), sperm strength (1-5) and morphology (%). Diluted semen samples were centrifuged at: 1500 g/10 min and “pellets” formed by sperm of each ejaculated, detached from the tube wall were diluted homogeneously in the diluent TRIS type up to the final volume of 1.5 mL. After that, packaged in 0.5 mL French straws, kept under refrigeration at 5ºC/4 h, placed in nitrogen vapor at -120ºC/15 min, and dipped in liquid nitrogen at -196ºC and then stored on identified rachis and stored in liquid nitrogen container until the time of thawing in  water bath at 37°C/30 s for semen microscopic analysis. Data from fresh, refrigerated and frozen semen were statistically analyzed by analysis of variance and the average compared by 5% of Tukey test. Fresh semen sperm concentration differed (P < 0.05) between the samples, rising after 40 days after the beginning of oral supplementation with selenium and vitamin E. For the spermatic strength, better score (P < 0.05) was observed at collection 4, in 40 days after the beginning of oral supplementation to dogs. For fresh and refrigerated semen, the total defects, defects of head, acrosome and tail did not differ (P > 0.05) between the samples. Total sperm defects and minor head and tail defects did not differ (P > 0.05) between the samples in post-thawing. Regarding the acrosome defects after thawing, there was a significant reduction (P < 0.05) in samples performed 40 and 60 days after the beginning of oral supplementation with selenium and vitamin E.Discussion: Attention should be paid for what purpose the extenders within the refrigeration or freezing biotech will be used. The managed supplement, by oral administration, containing selenium and vitamin E, influenced beneficially raising the sperm concentration in fresh semen and decreasing the acrosome defects in frozen semen. Oral administration of supplementation with selenium and vitamin E is recommended for improving the quality of fresh and frozen semen in dogs.

Effect of docosahexanoic acid on quality of frozen–thawed bull semen in BioXcell extender

2017 ◽  
Vol 29 (3) ◽  
pp. 490 ◽  
Author(s):  
Asmatullah Kaka ◽  
Wahid Haron ◽  
Rosnina Yusoff ◽  
Nurhusien Yimer ◽  
A. M. Khumran ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Optimum Level ◽  
Docosahexanoic Acid ◽  
Normal Morphology ◽  
Bull Semen ◽  
Acrosome Integrity

This study was conducted to investigate the effect of docosahexanoic acid (DHA) supplementation in BioXcell extender on the quality of frozen–thawed bull semen. Twenty-four ejaculates were collected from three bulls (eight from each bull). Ejaculates with motility ≥70% and normal morphology ≥80% were extended into BioXcell extender to which 0 (control), 3, 5, 10 or 15 ng mL–1 DHA was added. The supplemented semen samples were incubated at 37°C for 15 min for DHA uptake by spermatozoa. Later, samples were cooled for 2 h at 5°C and packaged into 0.25-mL straws, frozen in liquid nitrogen for 24 h and subsequently thawed for evaluation. Results are presented as percentages ± s.e.m. Supplementation with DHA at 3 ng mL–1 significantly improved sperm functional parameters including sperm motility, normal morphology, viability, acrosome integrity and membrane integrity when compared with other supplemented groups and the control. Lipid peroxidation increased as the incorporation of DHA supplementation increased. In conclusion, 3 ng mL–1 concentration of DHA resulted in superior quality of frozen–thawed bull spermatozoa and is suggested as the optimum level of DHA to be added into BioXcell extender.

scholarly journals Effect of fruit-juice on spermatozoa viability and lipid peroxidation of Red Sokoto bucks during liquid storage

2021 ◽  
Vol 41 (1) ◽  
pp. 16-27
Author(s):  
J. O. Daramola ◽  
T. A. Sorongbe ◽  
O. M. Onagbesan ◽  
A. V. Jegede ◽  
A. O. Ladokun ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Cell Damage ◽  
Egg Yolk ◽  
Protective Effects ◽  
Sperm Viability ◽  
Progressive Motility ◽  
Liquid Storage ◽  
Watermelon Juice ◽  
And Control

Antioxidants are linked with sperm viability because of their protective effects against cell damage during preservation. In order to enhance the life span of refrigerated buck semen, this study was carried out to determine the effect of fruit-rich antioxidants on spermatozoa viability and lipid peroxidation (LPO) of buck semen during liquid storage. Pooled semen from five Red Sokoto bucks was diluted with Tris-egg yolk based extender and supplemented each with juices from pawpaw tomato and watermelon at 0, 2.5, 5, 7.5 and 10/ 100 ml respectively. Following dilution, the semen samples were assessed subjectively after in vitro storage at 5°C for 24, 48, 72 and 96 hours as regards sperm motility, abnormalities, and acrosome status using a phase-contrast microscope. The concentration of malondialdehyde (MDA) as indices of lipid peroxidation (LPO) in the stored semen was measured in thiobarbituric acid reactive substances (TBARS) at 24, 48, 72 and 96 hours. The results showed highest progressive motility in watermelon juice at 2.5% (P<0.05) during the first 24 hours of storage while the lowest progressive motility was recorded at various levels of pawpaw juice (P<0.05). After 48 hours of storage, extender supplemented with watermelon and tomato juices had better progressive motility compared to control except 7.5% and 10%% of tomato juice (P<0.05). Irrespective of level of juice in the extender, the percentage of intact acrosome was similar among the various juices and control. The results showed that spermatozoa extended with watermelon juice had the lowest (P<0.05) percentage abnormality compared to other extenders at 24, 48, 72 and 96 hours of storage. Higher (P<0.05) percent spermatozoa abnormality compared to other fruit juices and control was observed at 72 and 96 hours of storage in spermatozoa extended with pawpaw juice. Significant reductions of MDA concentrations were achieved by addition of fruit-rich antioxidants to Tris-egg yolk based extender during the first 72 hours and the reduction was much pronounced in extender supplemented with pawpaw juice compared to control (P<0.05). The findings reveal that fruit-rich antioxidants from watermelon and tomato have protective ability to maintain sperm viability and to reduce concentration MDA of buck semen during liquid storage.

scholarly journals COMBINED EFFECT OF STREPTOMYCIN AND SWEET ORANGE ESSENTIAL OIL TO MEMBRANE AND ACROSOME INTEGRITY BOER GOAT FROZEN SEMEN

2019 ◽  
Vol 3 (2) ◽  
pp. 89
Author(s):  
Sukma Aditya Sitepu ◽  
Julia Marisa
Keyword(s):  
Essential Oil ◽  
Sweet Orange ◽  
Membrane Integrity ◽  
Boer Goat ◽  
Randomized Design ◽  
Frozen Semen ◽  
Acrosome Integrity ◽  
Completely Randomized Design ◽  
Orange Essential Oil

One of factors that cause a bad quality of Boer Goat frozen semen is the growth of bacterial. This can be overcome by adding antibiotics such as streptomycin. To further suppress the growth of bacteria can be added other ingredients that contain antibacterials such as sweet orange essential oil. The purpose of this research is to know the percentage value of Membrane Integrity and Acrosome Integrity on Boer Goat frozen semen with addition sweet orange essential oil and streptomycin. The method used was experimental using Completely Randomized Design with 5 treatments and 5 replications. The treatment in this research is addition 0%, 0,25%; 0.5%; 0.75% and 1% sweet orange essential oil on tris yolk and streptomycin extender. The results showed the best treatment addition combination streptomycin and sweet orange essential oil to percentage Membrane Integrity and Acrosome Integrity is increase 1% sweet orange essential oil.Keywords: Boer Goat, essential oil, frozen semen, streptomycin, sweet orange.

256 EFFECT OF SPERM SELECTION ON THE RATE OF IN VITRO FERTILIZATION IN ALPACA (VICUGNA PACOS)

2015 ◽  
Vol 27 (1) ◽  
pp. 217
Author(s):  
E. Mellisho ◽  
V. Rivas ◽  
J. Ruiz ◽  
G. Mamani
Keyword(s):  
Extended Period ◽  
Cumulus Cells ◽  
Sperm Selection ◽  
Cleavage Rate ◽  
Slow Cooling ◽  
Vitro Fertilization ◽  
Progressive Motility ◽  
Frozen Semen ◽  
Washing Method

In alpacas, improvement of reproductive efficiency of male camelids is limited by the small size of the testes, extended period of ejaculation, and low quality of semen. This study was designed to determine the effect of 2 sperm preparation treatments before IVF on the cleavage rate. The sperm was obtained by slicing the head of the epididymis of slaughtered male alpacas (n = 8), diluting in Tris-yolk-glycerol, and freezing with the slow-cooling method. Frozen semen straws per each male were thawed in a water bath at 37°C for 15 s and evaluated for percentage of progressive motility (32 ± 8.6%) and concentration (66.5 ± 24 × 106 sperm mL–1) post-thawing. Sperm selection by the swim-up method was performed by centrifugation at 1077 × g for 5 min with washing sperm medium eliminating the supernatant; sperm were settled in inclined tube with fertilization medium (without capacitating agent) for 60 min, after which 100 μL from the surface was recovered for use in IVF. The washing method consisted in repeated washing (twice) of sperm in washing sperm medium and fertilization medium by centrifugation at 1077 × g for 5 and 3 min, respectively, and recovery of 50 μL from the bottom of the tube for use in IVF. Sperm selected by swim-up or washing methods had similar characteristics of progressive motility (18 and 23%); however, the concentration was higher for the washing v. swim-up method (52 v. 14 × 106 sperm mL–1, respectively). Cumulus-oocyte complexes (COC) were recovered from 278 ovaries of alpacas killed at abattoirs and classified (Grade 1 and 2) for in vitro maturation (38.5°C at 5% CO2 in air for 27 h in 50 μL of 10 COC per drop). A total of 839 oocytes cultured for 27 h in maturation medium were partially stripped out of cumulus cells by gentle aspiration with a pipette. Sperm suspensions in Fert TALP medium (5 μL) from each treatment group were added to each fertilization drop with 10 oocytes per drop of 45 μL obtaining a final concentration of 10 × 106 sperm mL–1 and cultivated for 72 h until their evaluation. The data for the 13 repetitions of the rate of cleavage (2 to 8 cells) were converted to angular values (angle = arcsin √%) with the object of normalizing the distribution of the data; the analysis of variance was performed (complete randomised design with sub-sampling, P < 0.05) using SAS® version 8.0 for Windows. The rate of cleavage (cell division) did not show statistical differences (P = 0.67) for the swim-up method (37%; 155/421) v. washing method (35%; 147/418). The methods of sperm selection (swim-up and washing) did not affect the rate of IVF.

Rat lung microsomal lipid peroxidation: effects of vitamin E and reduced glutathione

1986 ◽  
Vol 61 (2) ◽  
pp. 785-790 ◽  
Author(s):  
D. P. Franco ◽  
S. G. Jenkinson
Keyword(s):  
Lipid Peroxidation ◽  
Vitamin E ◽  
Liver Microsomes ◽  
Sulfhydryl Group ◽  
Rat Lung ◽  
Active Components ◽  
Microsomal Lipid Peroxidation ◽  
Microsomal Membranes ◽  
High Concentrations

Lung microsomal membranes that contain the redox active components associated with the mixed-function oxidase system can be peroxidized in vitro. To investigate the characteristics of rat lung microsomal lipid peroxidation, we performed experiments using a variety of peroxidation initiators and microsomes obtained from normal and vitamin E-deficient rats. We found that lung microsomes obtained from normal rats are peroxidized much less than liver microsomes obtained from the same animals. Only initiation systems using very high concentrations of ferrous iron produced any significant peroxidation of normal rat lung microsomes. Lung microsomes obtained from vitamin E-deficient rats were found to be much more susceptible to peroxidation. Glutathione (GSH) was effective in inhibiting peroxidation when lung microsomes from normal rats were peroxidized. GSH was not effective in decreasing peroxidation when microsomes from vitamin E-deficient rats were peroxidized in the same system. We conclude that both GSH and vitamin E protect lung microsomal membranes from peroxidation. Glutathione protection appears to be related to the presence of a sulfhydryl group.

scholarly journals Cholesterol-loaded cyclodextrin plus trehalose improves quality of frozen-thawed ram sperm

2019 ◽  
Vol 64 (No. 03) ◽  
pp. 118-124 ◽  
Author(s):  
ME Inanc ◽  
S Gungor ◽  
C Ozturk ◽  
F Korkmaz ◽  
I Bastan ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Mitochondrial Membrane ◽  
Improve Quality ◽  
Progressive Motility ◽  
Computer Aided ◽  
Acrosome Integrity ◽  
Ram Sperm ◽  
Additive Control ◽  
Artificial Vagina

The objective of this study was to determine effects of supplementing Tris-based semen extenders with either cholesterol-loaded cyclodextrin (CLC) or 7-dehydrocholesterol loaded cyclodextrin (7-DCLC) plus trehalose (T) for cryopreservation of ram semen. Semen was collected with an artificial vagina from five Merino rams (2–3 years of age) during the non-breeding season. Ejaculates were pooled, divided into eight equal portions, diluted with a standard Tris-based extender containing: no additive (control); T (50 mM); or T (50 mM) + 1.5, 2.5 or 3.5 mg of either 7-DCLC or CLC. Semen was chilled from 37°C to 4°C, placed in 0.25 ml French straws, held 5 cm above liquid nitrogen for 12 minutes, then plunged into liquid nitrogen. After thawing, a computer-aided semen analyzer system (CASA) was used to assess motility, whereas plasma membrane and acrosome integrity (PMAI) and high mitochondrial membrane potential (HMMP) were assessed with flow cytometry. Sperm supplemented with 2.5 mg and 3.5 mg CLC + T had the highest (P &lt; 0.05) total and progressive motility (65.2 ± 4.7 and 19.0 ± 1.0% respectively, mean ± SEM), albeit with no significant differences from sperm with 1.5 or 3.5 mg CLC + T. Sperm with 2.5 mg CLC + T had the highest (P &lt; 0.05) PMAI (59.3%; not different from 3.5 mg CLC + T) and highest (P &lt; 0.05) HMMP (64.6%; not different from 1.5 or 3.5 mg CLC + T). The lowest ALH value, 2.8 ± 0.3 µm was in the 2.5 mg 7-DCLC + T group; otherwise, there were no significant differences among groups for any other CASA end point. In conclusion, adding CLC + T to a tris-based extender optimized quality of frozen-thawed ram semen. Therefore, extenders including CLC + T have potential to improve quality of frozen-thawed ram sperm.

scholarly journals Relationship between tissue lipid peroxidation and peroxidizability index after α-linolenic, eicosapentaenoic, or docosahexaenoic acid intake in rats

2003 ◽  
Vol 89 (1) ◽  
pp. 19-28 ◽  
Author(s):  
Morio Saito ◽  
Kazuhiro Kubo
Keyword(s):  
Lipid Peroxidation ◽  
Vitamin E ◽  
Docosahexaenoic Acid ◽  
Nutritional Status ◽  
Dietary Vitamin ◽  
Tissue Lipid ◽  
Double Bonds ◽  
Adequate Level ◽  
Peroxidizability Index ◽  
Tissue Lipid Peroxidation

In a previous study, we found that the extent of dietary n-3 docosahexaenoic acid (DHA)-stimulated tissue lipid peroxidation was less than expected from the relative peroxidizability index of the total tissue lipids in rats with adequate vitamin E nutritional status. This suppression of lipid peroxidation was especially prominent in the liver. To elucidate whether this phenomenon was unique to DHA, we compared the peroxidation effects of n-3 α-linolenic acid (α-LN) and n-3 eicosapentaeonic acid (EPA) with those of DHA in rats. Either α-LN (8·6 % of total energy), EPA (8·2 %), or DHA (8·0 %) and one of two levels of dietary vitamin E (7·5 and 54 mg/kg diet) were fed to rats for 22 d. Levels of conjugated diene, chemiluminescence emission and thiobarbituric acid (TBA)-reactive substance in the liver, kidney, and testis were determined as indicators of lipid peroxidation. In rats fed the DHA diet deficient in vitamin E (7·5 m/g diet), TBA values in the liver, kidney, and testis correlated well with the tissues' relative peroxidizability indices. In rats fed the α-LN diet with an adequate level of vitamin E (54 m/g diet), a close association between relative peroxidizability indices and lipid peroxide levels was observed in all the tissues analysed. However, in rats fed either the EPA diet or the DHA diet with an adequate level of vitamin E, the extent of lipid peroxidation in each tissue was less than expected from the relative peroxidizability index. This suppression was particularly marked in the liver. We concluded that suppression of lipid peroxidation below the relative peroxidizability index was not unique to DHA, but was also seen with EPA, which has five double bonds, in rats with adequate vitamin E nutritional status, but not with α-LN, which has three double bonds.

Vitamin E Reduces Monocyte Tissue Factor Expression in Cirrhotic Patients

Blood ◽  
1999 ◽  
Vol 93 (9) ◽  
pp. 2945-2950 ◽  
Author(s):  
Domenico Ferro ◽  
Stefania Basili ◽  
Domenico Praticó ◽  
Luigi Iuliano ◽  
Garret A. FitzGerald ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Liver Cirrhosis ◽  
Vitamin E ◽  
Tissue Factor ◽  
Standard Therapy ◽  
Pathophysiological Mechanism ◽  
Important Mediator ◽  
Factor Expression ◽  
Cirrhotic Patients

Clotting activation may occur in liver cirrhosis, but the pathophysiological mechanism has not been fully elucidated. Because a previous study demonstrated that lipid peroxidation is increased in cirrhosis, we analyzed whether there is a relationship between lipid peroxidation and clotting activation. Thirty cirrhotic patients (19 men and 11 women; age, 34 to 79 years) and 30 controls matched for sex and age were investigated. In all subjects, monocyte expression of tissue factor (TF) antigen and activity; plasma levels of prothrombin fragment 1+2 (F1+2), a marker of thrombin generation; and urinary excretion of Isoprostane-F2-III, a marker of lipid peroxidation, were measured. Furthermore, the above-reported variables were re-evaluated after 30 days of treatment with standard therapy (n = 5) or standard therapy plus 300 mg vitamin E twice daily (n = 9). In addition, we analyzed in vitro if vitamin E (50 μmol/L) influenced monocyte TF expression and F1+2 generation. Cirrhotic patients had higher values of Isoprostane-F2-III (P < .0001), F1+2 (P < .0001), and monocyte TF antigen (P < .0001) and activity (P < .03) than controls. Isoprostane-F2-III was significantly correlated with F1+2 (Rho = 0.85; P < .0001) and TF antigen (Rho = 0.95; P < .0001) and activity (Rho = 0.94;P < .0001). After vitamin E treatment, Isoprostane-F2-III (P = .008), F1+2 (P < .008), and monocyte TF antigen (P = .012) and activity (P = .008) significantly decreased; no changes of these variables were detected in patients not receiving vitamin E. In vitro, vitamin E significantly reduced the expression of monocyte TF antigen (−52%; P = .001) and activity (−55%; P= .003), as well as F1+2 generation (−51%; P = .025). This study shows that vitamin E reduces both lipid peroxidation and clotting activation and suggests that lipid peroxidation may be an important mediator of clotting activation in liver cirrhosis.

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