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Relation between respiratory activity and sperm parameters in boar spermatozoa cryopreserved with alpha‐tocopherol and selected by Sephadex
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Our aim was to evaluate the effect of Sephadex filtration on respiratory activity of porcine spermatozoa and its relation with quality and functional sperm parameters. Samples were evaluated regarding oxygen uptake and sperm parameters: motility, plasma and acrosome membrane integrity, capacitation and acrosome reaction induction in vitro, plasma membrane functionality, determined by the hypo‐osmotic swelling test (HOST), and lipid peroxidation assessed by thiobarbituric acid assay. Sephadex filtration improved all routine quality parameters (motility, plasma and acrosome membrane integrity) and functional parameters (HOST, in vitro capacitation and true acrosome reaction levels) and produced a significant decrease in cryocapacitation and lipid peroxidation. Oxygen uptake increased in Sephadex samples (41 ± 7%) respect to single washing. Oxygen addition of carbonyl‐cyanide‐m‐chlorophenylhydrazone (CCCP) confirmed mitochondrial coupling in washed and Sephadex samples; showing an increase of 2.6 and 4.2 times for oxygen consumption in single washing and Sephadex ones, respectively. The increase in oxygen uptake with succinate addition with respect to basal oxygen uptake was significantly lower in Sephadex samples (63 ± 25%) than in the washed ones (183 ± 35%). Sephadex samples showed higher mitochondrial activity measured by oxygen consumption and improved quality and functional parameters. Our study recommends this protocol due to the fact that this filtration method removes dead or damaged spermatozoa allowing to obtain cryopreserved boar spermatozoa with optimized fertilizing capacity. 相似文献
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HT Duong DJ Skarzynski KK Piotrowska‐Tomala MM Bah K Jankowska P Warmowski K Łukasik K Okuda TJ Acosta 《Reproduction in domestic animals》2012,47(6):939-945
Previous in vitro studies demonstrated that bovine endometrium has the capacity to convert inactive cortisone to biologically active cortisol (Cr) and that Cr inhibits cytokine‐stimulated prostaglandin F2α (PGF) production. This study was carried out to test the hypothesis that bovine reproductive tract has the capacity to convert cortisone to Cr in vivo and to evaluate the effects of intravaginal application of exogenous cortisone on uterine PGF secretion during the late luteal stage. The temporal relationships between PGF and Cr levels in uterine plasma were also determined. Catheters were inserted into jugular vein (JV), uterine vein (UV), vena cava caudalis (VCC) and aorta abdominalis (AA) of six cows on Day 15 of the oestrous cycle (ovulation = Day 0) for frequent blood collection. On Day 16, the cows were divided randomly into two groups and infused intravaginally with vaseline gel (10 ml; control; n = 3) or cortisone dissolved in vaseline gel (100 mg; n = 3). Blood samples were collected at ?2, ?1, ?0.5, 0, 0.5, 1, 1.5, 2, 3, 4, 5 and 6 h after treatments (0 h). Intravaginal application of cortisone increased plasma concentrations of Cr between 0.5 and 1.5 h in UV, at 0.5 h in VCC, at 1 h in JV and at 1.5 h in AA. The plasma concentrations of PGF in UV and of PGF metabolite in JV were greater at 0.5 and 1 h in the cortisone‐treated animals than in control animals. The levels of PGF in UV blood plasma decreased after Cr reached its highest levels. The overall findings suggest that the female reproductive tract has the capacity to convert cortisone to Cr in vivo. Based on the temporal changes of PGF and Cr levels in the uterine plasma, a biphasic response in PGF secretion was found to be associated to the Cr increase induced by the cortisone treatment at the late luteal stage in non‐pregnant cows. 相似文献
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E Bussalleu E Pinart MM Rivera M Briz S Sancho M Yeste I Casas A Fàbrega T Rigau JE Rodriguez-Gil S Bonet 《Reproduction in domestic animals》2009,44(3):499-503
The aim of this work was to develop a method to enhance the sperm parameters of ejaculates with low sperm quality from Piétrain boars. Seminal doses were filtered through columns of DEAE Sephadex (length 2.5 ± 0.5 cm), CM Sephadex (length 5 ± 0.5 cm), glass wool (length 2 ± 0.5 cm) or glass bead (length 10 ± 0.5 cm), with an exit flow rate of 1 ml/40 s in all cases. For each male, 10 ml of the sperm cell-rich fraction diluted at 1 : 6 were filtered. Sperm quality was assessed before and after filtration. Sperm morphology, sperm motility and sperm concentration were determined using the computer program sca ® 2002 Production, and sperm viability was evaluated by fluorescence multistaining. Osmotic resistance test and hyperosmotic resistance test were used to determine the osmotic resistance of spermatozoa, whereas l -lactate production estimated the metabolic activity. Results showed a decrease of sperm concentration and osmotic resistance of spermatozoa after filtration in the four matrixes. However, an increase in the frequency of viable spermatozoa with intact acrosome after filtration in glass bead columns and an increase of morphologically normal spermatozoa after filtration in Sephadex CM-50, glass wool and glass bead columns were observed. Despite the decrease in the frequency of progressive motile spermatozoa, l -lactate production and mitochondrial sheath integrity maintained constant after filtration. Our findings indicate that column filtration is an effective method to enhance the sperm quality by selecting viable and morphologically normal spermatozoa without altering DNA, plasma membrane, mitochondrial sheath integrity or inducing premature acrosome reaction. 相似文献
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The effects of stale seedbed preparations and several weed control methods on the emergence of weeds in lettuce were studied. The specific goal was to evaluate the use of a stale seedbed in combination with chemical or mechanical weed control methods in the field. Depending on location and year, stale seedbed preparations followed by weed control prior to planting reduced the amount of weeds during crop growth by 43–83%. Control of the emerged seedlings after a stale seedbed preparation was more effective with glyphosate than with a rotary harrow. Covering the rotary harrow during control to prevent light reaching the soil improved its effect on the weed density during crop growth in two of 3 years. Radiation with far red light (FR) did not reduce the number of emerging weeds in this study. Mechanical control by finger weeder, torsion weeder and hoe was applied without stale seedbed preparations. These measures reduced the weed densities by 88–99%, compared with the untreated control and were more effective than chemical weed control with carbetamide and chlorpropham. The results show that the stale seedbed technique in combination with mechanical control of emerging weeds can reduce the weed population during crop growth as effectively as chemical control. The technique may therefore help reduce the use of herbicides in lettuce crops in the future. 相似文献
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Clay A. Cavinder PhD Martha M. Vogelsang PhD PAS Pete G. Gibbs PhD PAS David W. Forrest PhD PAS David G. Schmitz DVM 《Journal of Equine Veterinary Science》2007,27(2):72-79
The influence of progesterone, luteinizing hormone (LH), and estrogen on the mare's estrous cycle has been well researched and documented, but other endocrine profiles have not received as much attention. To evaluate endocrine concentrations in fat-conditioned (body condition score [BCS] of 7–8) versus moderately conditioned mares (BCS of 5–6), 24 mares were allotted to and maintained in respective groups from late gestation until pregnancy was confirmed after breeding on the second postpartum estrus. Serum concentrations of thyroxine (T4), insulin-like growth factor-1 (IGF-1), and leptin were assayed to characterize circulating blood concentrations. Additionally, LH and progesterone serum concentrations were assayed to evaluate the estrous cycle status of the mares. Leptin and progesterone concentrations were not different (P > .05) between the groups. Nevertheless, serum concentrations of T4 were higher (P < .01) and IGF-1 concentrations lower (P < .01) in moderately conditioned as compared with fat-conditioned mares during times of ovulation and the interovulatory period. Furthermore, serum concentrations of LH were found to be different between the groups only when the estrous cycle approached the second ovulation (P < .0001). Results of this study suggest that mares maintained in a BCS of 5 or greater are similar in terms of reproductive efficiency. Although the circulating serum concentrations of T4 and IGF-1 are different after parturition, their influence does not affect reproductive capabilities of mares with a BCS of 5 or greater. 相似文献
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J Miró A Flotats MM Rivera M Ocaña E Taberner A Peña T Rigau 《Reproduction in domestic animals》2006,41(S2):103-103
In order to determine the injure produced in boar spermatozoa through cryopreservation process, we analyzed the expression of the hexose transporters Glut-3 and Glut-5 and the zona pellucida binding protein As-A (P68) in three different steps of the freezing-thawed protocol: at 17°C (fresh BTS-diluted semen, 1 : 2 v/v, step 1), at 5°C (after glycerol addition; step 2), and post-thawing (step 3). All sperm analyses were carried out with immunogold techniques under electronic microscopy. For this study eight healthy post-pubertal Iberian boars were submitted to a collection of twice per week through 3 months, evaluating two ejaculates from each boar. Glut-3 maintains the expression in the acrosome region post-thawing but not along the tail where is reduced. The expression of Glut-5 and As-A is majority located at the post-acrosome region of the spermatozoa at step 1, but in step 2 and step 3 this expression is relocated to sperm tail area. In conclusion, while cryopreservation affects the localization and the expression of Glut-3 and Glut-5, its fertilizing capacity is not significantly reduced. The stabilization of boar semen at 5°C was found to be the most crucial step for sperm survival. 相似文献