Effect of Aqueous Extract of Maca Addition to an Extender for Chilled Canine Semen (original) (raw)
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Theriogenology, 2007
The objective of this study was to evaluate the quality of extended dog semen processed with diluents containing various concentrations of vitamin C. Ejaculates from five dogs were collected, pooled and evaluated for concentration, sperm motility, rapid steady forward movement (RSF-movement), viability, acrosomal integrity and by the hypo-osmotic swelling test. Also, superoxide (O 2 À ) production, hydroxyl radicals (OH ) and total reactive oxygen species (tROS) were determined. The pool was divided in five aliquots, which were diluted to a final concentration of 66 Â 10 6 spermatozoa/ml with a Tris-glucose-egg yolk extender containing one of the following concentrations of vitamin C (0, 0.1, 0.5, 1 or 2.5 mM). The semen aliquots were chilled and preserved at 4 8C. Portions of chilled semen were removed at 24 and 72 h, and semen quality was evaluated after rewarming. This process was repeated 10 times in pooled semen of the same origin and data were analysed by one-way analysis of variance. At both times, none of the semen quality parameters were positively influenced ( p > 0.05) by vitamin C supplementation. At 24 h, none of the reactive oxygen species (O 2 À , OH , tROS) were significantly altered. At 72 h, significant reductions of O 2 À production were observed by the concentrations of 0.1, 0.5 and 2.5 mM compared with the 0 mM concentration ( p = 0.049). Also, at 72 h, the 2.5 mM concentration showed significantly lower OH values in comparison with the control group ( p = 0.048). In conclusion, addition of vitamin C to semen extenders does not benefit the quality of canine extended spermatozoa. #
Advances in Animal and Veterinary Sciences, 2023
During chilled storage oxidative stress is the primary cause for the reduction in the longevity of canine sperm. To prolong the chilled canine sperm quality, the study investigated the effects of the adding antioxidants to semen extender on canine sperm during 12 days of chilling storage. Twenty ejaculates from five American Bully dogs were used. The sperm quality was determined by system automatic sperm analyser for sperm motility parameters, and by confocal laser scanning microscope for sperm membrane integrity parameters. Sperm lipid peroxidation was also analysed through malondialdehyde production. The study found that there was a gradual decrease in all sperm quality indicators of all the treatments during 12 days of storage. In addition, the chilled sperm motility parameters among all the semen extenders were not noticeably different at each time period of evaluation (P>0.05). Although sperm lipid peroxidation index in the Ocimum gratissimum essential oils treatment was higher than those in the treatment of vitamin E and green tea polyphenols, the percentage of the high mitochondrial membrane potential, the intact plasma membrane and acrosome membrane indicators in the Ocimum gratissimum essential oils treatment were higher than those in the rest treatments and had a substantial difference when compared to the control treatment (P<0.05). In concluding, semen extender supplemented with Ocimum gratissimum essential oils is appropriate for improving the chilled canine sperm for the duration of 12 days in storage.
Journal of veterinary research, 2018
The addition of low-molecular-weight antioxidants during the freezing process improves post-thaw sperm quality. The high antioxidant potential of cryopreserved semen could have a positive effect on the motility, viability, and energy status of sperm cells and their ability to bind to the of oocytes. The aim of the study was to determine the effects of different concentrations and combinations of vitamins E and C in a semen extender on selected quality parameters of frozen-thawed canine spermatozoa. The experimental material was the semen of four mixed-breed dogs. Sperm viability (motility, plasma membrane integrity, and mitochondrial function) was examined at 0, 60, and 120 min in semen samples supplemented with the extender and in the controls. Combined supplementation with vitamins C + E at a concentration of 200 + 200 μM /1 × 10 spermatozoa had the most profound effect on total sperm motility, linear motility, and the percentage of spermatozoa with intact plasma membrane and acti...
Cryo letters
Vitamin C and green tea polyphenol are known to have antioxidant effects. The aim of this study was to evaluate the quality of canine semen after preservation with diluents containing vitamin C and polyphenol at 5 degree C for 4 weeks. In experiment 1, we investigated the effects of vitamin C combined with polyphenol supplementation on chilled semen quality. The addition of vitamin C (0.5 or 1 mM) with 0.75 mg per mL polyphenol to semen extender provided significantly higher percentages of sperm motility and viability during cold storage compared to unsupplemented semen. In experiment 2, we determined the optimal working concentration of vitamin C in the semen extender by comparison of a range of concentrations between 0.1 and 20 mM. Supplementation of 0.5 mM vitamin C plus polyphenol yielded the highest percentages of sperm motility and viability; however, there was no beneficial effect on the plasma membrane and acrosomal integrity of the spermatozoa.
Journal of Veterinary Science, 2018
This study investigated the correlation between oxidative stress status and key canine sperm parameters and the effect of addition of a superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) combination in egg yolk tris-citrate glucose (EYT-G) extender on semen during 10 days of storage at 4 o C. Ten Boxer dogs were divided into two groups, fertile (F) and hypofertile (H), depending on pregnancy and live birth rate status in the previous year. Semen evaluation was performed on the day of collection (D0) and after 5 (D5) and 10 (D10) days of cooled storage. Sperm motility, kinetic parameters, and DNA integrity were assessed. A correlation between oxidative status and key semen parameters in both F and H groups was observed. Total and progressive motilities were significantly higher in the treated (SOD, CAT, and GPx addition) versus control groups at D10 in both F and H groups, and at D5 in the H group. DNA integrity was significantly higher in both treated groups (H and F) at D5 and D10. In conclusion, the addition of SOD, CAT, and GPx in the extender allows preservation of semen quality for up to 10 days of storage at 4 o C in both fertile and hypofertile dogs.
2016
The use of antioxidants as semen additives in the extender has been proposed to improve sperm function during cool storage. The objective of this study was to compare the effects of different antioxidant supplementation in extender on boar semen quality during storage for 10 days at 17oC. Semen samples (n=18) were divided into 10 aliquots and diluted with Reading extender supplemented with three different concentrations of water-soluble glutathione, Lcarnitine, and vitamin E analogue Trolox (0.1, 1 and 5mM). All samples were stored in a refrigerator at 17oC for 10 days. Sperm motility was assessed at Days 1, 4, 7 and 10 of storage under a phase-contrast microscopy. Percentage of live sperm with normal apical ridge and high mitochondrial energy status was assessed at Days 1, 5 and 10 of storage by fluorescent multiple staining using propidium iodide (PI), fluorescein isothiocyanate-labeled peanut (Arachis hypogaea) agglutinin (FITC-PNA) and 5,5’,6,6’-tetrachloro-1,1’,3,3’ tetraethylb...
Anti-oxidative Status and Semen Quality during Cooled Storage in Stallions
Journal of Veterinary Medicine Series A, 2006
Activity of the anti-oxidative enzymes glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT), content of thiobarbituric acid reactive substances (TBARS) and SH-groups were determined in native stallion semen (n = 8 stallions). Semen was then diluted in Kenney extender, EquiPro((R)) extender either with or without addition of N-acetyl cysteine or phosphate-buffered saline (PBS) and stored for 72 h at 5 degrees C. Correlations between initial activity of enzymes and development of semen motility and membrane integrity were calculated. Activities of GSH-Px, SOD and CAT immediately after semen collections were 10.0 +/- 0.6 picokatals, 0.40 +/- 0.03 SOD units and 0.70 +/- 0.05 nanokatals/10(6) spermatozoa respectively. TBARS content was 0.06 +/- 0.01 nmol and SH-group content 1.7 +/- 0.5 mmol/10(6) spermatozoa. The loss of motile spermatozoa during storage did not differ between extenders. N-acetyl cysteine had no effect on semen motility and membrane integrity. The loss in membrane-intact spermatozoa was highest (P < 0.05) in semen diluted in PBS. Motility and membrane integrity after addition of extender were positively correlated with GSH-Px and CAT, indicating that anti-oxidative mechanisms contribute to the initial high percentage of motile and membrane-intact spermatozoa. However, in these samples the decrease in semen quality was most pronounced. No correlations existed between initial activity of anti-oxidative enzymes, peroxidation products and semen quality during storage. This indicates that once extender has been added, peroxidative damage to sperm membranes is not the predominant cause of losses in semen quality.
2017
The effects of adding various concentrations of antioxidant, butylated hydroxytoluene (BHT) on chilled bull semen for 72 h at 4°C in Bioxcell® (BX), Tris egg-yolk- (TEY) and citrate egg-yolk- (CEY) based semen extenders were studied. Twenty-two ejaculates collected from three bulls, each extended using three extenders with BHT at 0 (control), 0.5, 1.0, 1.5, 2.0 and 3.0 mM/mL, were evaluated for sperm quality parameters. Extended semen was packaged into 0.25 mL straws containing 20 x 106 spermatozoa and chilled for 72 h. Four random straws each from the control and treatment groups were warmed at 37°C, pooled and evaluated using a computer-assisted semen analyser (IVOS Hamilton Thorne Biosciences) for general and progressive motilities, and for morphology, viability and acrosome damage using eosin-nigrosin stain under phase-contrast microscope. Results showed that sperm morphology, viability and protection of acrosome damage were significantly improved (p<0.05) at BHT concentratio...
International Journal of Andrology, 2018
Stallion semen is damaged by oxidative stress during cooling and transport. Semen processing and extenders have been tested to improve the fertilizing capacity of semen and to preserve semen during transport. Dietary supplementation with natural antioxidants has been proposed to prevent oxidative damages. In this study, for the first time, the effect of dietary supplementation with Lepidium meyenii (Maca) on the characteristics of fresh and chilled stallion semen was evaluated. Maca is a traditional Andean crop used as a nutraceutical for the fertility-enhancing properties that are linked with antioxidant activity. The diet of five stallions was supplemented with 20 g of Maca powder daily for a total of 60 days. A control group of five stallions received the same diet without Maca. Semen was collected once before the administration of Maca (D0), twice during the administration at 30 and 60 days (D30 and D60), and finally twice at 30 and 60 days after the end of the administration (D90 and D120). Ejaculates were processed for cooled shipping at 5°C and evaluated in the laboratory for total and progressive motility, acrosome integrity, and lipid peroxidation after collection and after 24, 48, and 72 h of storage. Dietary supplementation with Maca improved sperm concentration (from 213 AE 80.4 to 447 AE 73.1 9 10 6 spz/mL) and total sperm count (from 10,880 AE 4377 to 24,783 AE 4419 9 10 6 spz). The beneficial effects of Maca supplementation on motility and acrosome integrity in the raw semen were detected from the end of treatment with Maca (D60) until the end of the study (D120). Furthermore, during cooling storage, total motility, progressive motility, and acrosome integrity declined more slowly in the Maca-treated group than in the control group. Lipid peroxidation did not change during cooling storage in either group and did not show a significant difference between the two groups. In this study, the dietary supplementation with Maca increased sperm production and stabilized semen quality during chilled storage.
The effect of inclusion of three antioxidants (Vitamin E, cysteine and glutathione) in ram semen extender on the release of antioxidant enzymes and free radicals was studied. A 3 × 3 factorial experiment was conducted to test effects of supplementing ram semen extender with Vitamin E (1, 5 and 10 IU), cysteine (1, 5 and 10 mM) or glutathione (0.5, 1 and 2 mM) on the sperm survival and release of superoxide dismutase (SOD), glutathione peroxidase (GPX) and thiobarbituric acid reactive substances (TBARS). Eighty ejaculates of eight fertile Najdi rams were collected, assessed for the gross examination and the good ejaculates (≥90% motility) were pooled and sperm count was assessed. Therefore ten extenders; control (C) and 9 treated (C plus antioxidant) were tested. Extended semen was stored at 5˚C for 96 h, examined for motility and survival and sperm cells were separated from plasma, sonicated, homogenized and exposed to the determinations of SOD, GPX and TBARS. The highest sperm survival was found in diluents containing 5 IU Vitamin E/ml, 1 and 2 mM glutathione (55.5% survival), while the lowest survival was found in 10 mM cysteine (11.1%). TBARS concentration was highest (P < 0.05) in control than other treatments, however, Vitamin E and glutathione exhibited low values. Contrariwise, activities of SOD and GPX increased (P < 0.05) within sperm cells and seminal plasma in diluents containing 5 IU E, 1 mM and 2 mM glutathione. The enzymatic activities were generally higher in seminal plasma than in sperm cells. It was concluded that supplementing ram semen extender during chilled storage with 5 IU Vitamin E per ml or 1 - 2 mM glutathione enhanced sperm survival and reduced free radicals.