Silencing the triacylglycerol lipase (TGL) gene decreases the number of apyrene sperm and inhibits oviposition in Sitotroga cerealella
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Spermatophore
Sitotroga cerealella
ABSTRACTObjective: This study aimed to determine the relationship between the addition of Glutathione to sperm freezing medium and cryosurvival rate of sperm motility and viability in asthenozoospermic patients.Methodology: fifty nine semen samples of asthenozoospermic patients were obtained from the male partner of infertile couples attending the fertility center of al-sadder medical teaching city in Al-Najaf during the period 1st October 2016 until 1st March 2017.Each semen sample was divided into four equal parts, volume of each part is 0.5 ml: first part (before activation), second part (after activation), third part (cryopreservation with sperm freezing medium for one month), and fourth part (cryopreservation with sperm freezing medium plus glutathione for one month ) .Sperm concentration , sperm progressive motility, sperm viability and sperm normal morphology were counted in first two parts, cryosurvival rate of sperm motility, viability was counted in third and fourth parts .Statistical analysis of the study performed by using Independent t-test and paired test to assess the differences between two groups .Results: After sperm activation by using the swim up technique ,the study results showed significant increase (P < 0.01) of progressive sperm motility, normal sperm morphology and sperm viability, while there were a significant decrease (P < 0.01) of sperm concentration. cryosurvival rate of sperm motility and viability were significant decrease (p < 0.01) after cryopreservation and thawing processes while the results were revealed significant improvement in cryosurvival rate of sperm motility, and viability ( p < 0.01 ) after the addition of glutathione to the sperm freezing medium.Conclusion: it can conclude from this study that addition of glutathione to sperm freeze medium improves. (p < 0.01) cryosurvival rate of sperm motility and viability during cryopreservation and thawing processes . Recommendations: the study recommended to addition 50 microliter of glutathione to sperm freezing media in concentration 1 mM .
Semen Analysis
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Female sperm storage
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Factors controlling sperm production in members of the Orthoptera have not been fully elucidated. In this study, the influence of intermating interval and ad libidum mating on sperm number was investigated in black field crickets (Teleogryllus commodus). Remating at a variety of time intervals was not characterized by a significant change in sperm number compared with the first mating. Ad libitum mating (i.e., continuous availability of unmated females) had two main effects on spermatophore production and sperm number: first, there was a trend toward increased time between copulations with each successive remating, and second, the number of spermatozoa encapsulated in the transferred spermatophore declined after most rematings, with 61.8% of the initial sperm number being produced for the second spermatophore and 51.3% of the initial sperm number being produced for the third. The decrease in mean sperm number was accompanied by increased variance in sperm number in later rematings. This study suggests that males are willing to suffer a decrease in sperm number if a mating opportunity occurs before the completion of sperm production.
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Evaluation of rat sperm by flow cytometry: Simultaneous analysis of sperm count and sperm viability.
In this study, we conducted a simultaneous analysis of sperm count and viability in rats by flow cytometry (FCM). Epididymal fluids were taken from the caudal epididymis of 12 to 13 week-old Sparague-Dawley rats. The fluids were weighed and mixed with Dulbecco's phosphate buffered saline (D-PBS). Propidium iodide, which can stain only dead sperm, was used to distinguish viable and dead sperm. The sperm count and viability analyzed by FCM were 1.28×106/mg and 78.0%, respectively. These values were consistent with the corresponding values (1.39×106/mg and 81.0%) that were directly determined microscopically in the fluids of the same sample. In addition, when the original mixture containing sperm was diluted two times and four times with D-PBS, or was diluted two times with D-PBS containing only killed sperm, the sperm count and viability determined by FCM also correlated well with the sperm count (r=0.96, P<0.01) and sperm motility (r=0.99, P<0.01) by direct microscopic observation, respectively. In conclusion, the present flow cytometric analysis would be practical for the simultaneous determination of sperm count and viability in rat epididymal fluids.
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Villosa
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Six-week old SD-Slc male rats were treated for 4 weeks with compounds known to induce toxicological changes in male reproductive organs (pyridoxine in saline, 500 mg/kg/day, i.p.) or sperm (trimethylphosphate in distilled water, 100 mg/kg/day, p.o.). Each sperm sample taken from the cauda epididymis was analyzed with flow cytometry for the evaluation of sperm viability and counts. Sperm motility and morphology by microscopical observation, and histopathological examination of reproductive organs were also performed for estimating the adverse effects of each compound on spermatogenesis and sperm. While a decrease in sperm motility was noted for the trimethylphosphate group, the low motile sperm was evaluated as being viable sperm, not moribund, with flow cytometry. In the pyridoxine group, microscopical observation revealed morphological changes of sperm and a decrease of motility. The present sperm analysis with flow cytometry also suggested morphological changes reflected by dot plot as well as decrease of sperm viability and counts. These results indicated that this procedure led to profound findings in the compound-treated animals, with evaluation as viable sperm, not moribund, in a low motile sperm sample, and suggesting morphological changes in the dot plot of flow cytometry.
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Viable, healthy sperm are preferred for oocyte fertilization with intracytoplasmic sperm injection. Currently, motility is the most widely applied measure of sperm viability. However, with this criterion, viable but immotile sperm are overlooked as candidates for micromanipulation. Supravital stains identify viable sperm but may be toxic to the gamete or embryo. We tested the hypothesis that hypo-osmotic swelling, developed to assess sperm membrane integrity, can accurately determine sperm viability. Thawed sperm from 12 fertile donors were exposed to a hypo-osmotic solution and to two supravital stains. A total of 2,010 sperm were assessed for tail coiling after a 30-min exposure to hypoosmotic solution. By supravital stains, 31% of thawed sperm were viable; 23% showed tail coiling. Among coiled-tail sperm, 100% were viable by supravital stain. As a measure of viability, tail coiling exhibited a sensitivity of 30%, specificity of 100%, and positive predictive value of 100% compared to supravital stains. With a 60-min hypo-osmotic incubation, the specificity (89%) and positive predictive value (78%) decreased significantly (P < 0.05). Therefore, hypo-osmotic swelling accurately detects viable sperm among a nonmotile population. Assay accuracy, however, is very sensitive to the incubation time in hypo-osmotic solution.
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The parasitoid Pteromalus cerealellae (Ashmead) was introduced at densities of one, five, or 10 pairs into 0.47-liter jars containing 500 g of corn that had been infested with one, five or 10 pairs of the Angoumois grain moth, Sitotroga cerealella (Olivier), adults. The abundance of S. cerealella progeny was significantly reduced by all release rates of parasitoids and at all densities of S. cerealella. As release rates increased from one to 10 pairs of P. cerealellae, the average percentage suppression increased from 85.5 to 99.2 at one pair of S. cerealella, from 77.2 to 98.0 at five pairs of S. cerealella, and from 66.9 to 95.8 at 10 pairs of S. cerealella. However, the degree of suppression was significantly influenced by both the numbers of P. cerealella released and the initial density of S. cerealella. With greater host density, more progeny of P. cerealellae were produced, but releasing more parasitoids did not significantly increase the number of P. cerealellae progeny.
Sitotroga cerealella
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