Examinando por Materia "Seminal plasma"

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Cooling of alpaca spermatozoa using an extender with the addition of different percentages of seminal plasma
(Elsevier, 2023-12-06) Bertuzzi, Mariana Lucía; Torres Mamani, Edita Yola; Pérez Durand, Manuel Guido; Huanca Mamani, Teodosio; Giuliano, Susana María; Carretero, María Ignacia
The objective of this study was to evaluate the effect of the addition of different percentages of seminal plasma (SP) during the cooling at 5 °C of alpaca spermatozoa from vas deferens. Fifteen pools of sperm from vas deferens were evaluated and then divided into four aliquots that were diluted to a final concentration of 30 × 106 sperm/ml with either: (1) Tris with 20% egg yolk (T-EY) (control, 0% SP), (2) T-EY with 10% SP, (3) T-EY with 25% SP, and (4) T-EY with 50% SP. Samples were cooled at 5 °C and the following sperm parameters were evaluated after 24 and 48 h of storage: motility, viability, membrane function, acrosome integrity, morphology, and chromatin condensation. Motility was also evaluated after 72 h of storage. A significant decrease in progressive and total sperm motility was observed in samples cooled with 50% SP with respect to all diluted samples, while these parameters were preserved in samples cooled with 0%, 10%, and 25% SP. The percentages of sperm viability, normal morphology, and highly condensed chromatin did not change after the cooling process and were similar between cooled samples. Although a significant decrease was observed in the percentage of spermatozoa with functional membranes and with an intact acrosome in all refrigerated samples compared to raw sperm, the greatest decrease was observed in samples cooled with 50% SP. No advantage was observed from the addition of SP to alpaca spermatozoa obtained from vas deferens and being cooled. In addition, to preserve the sperm motility of cooled samples for up to 72 h, it should be recommended to include a 10% SP in the extender.
Effect of seminal plasma on hipoosmotic swelling test in fresh alpaca spermatozoa
(MedCrave, 2017-12-22) Pacheco Curie, Joel I.; Mamani Cato, Rubén Herberht; Vélez, V. M.
A study was designed with the objective of evaluating the effect of seminal plasma on the response to the hypoosmotic swelling test (HOST) in alpaca spermatozoa, for which three experimental groups were organized as follows: Group 1(n=15) plasma free sperm seminal (obtained from the vas deferens, aspirated in PBS), Group 2(n=15) free seminal plasma sperm reconstituted with seminal plasma (obtained from the vas deferens, aspirated in PBS, mixed in 50/50% with seminal plasma) and Group 3(n=15) whole semen (obtained by artificial vagina), The samples were incubated in a hypoosmotic solution adjusted to 100mOsmol (sodium citrate+fructose+2H2Ocsp 100mL). 0.1mL of semen+0.9mL of hypoosmotic solution was mixed, incubated for 30minutes in a water bath at 37°C and the reaction was stopped with 0.1mL of 4% formaldehyde. A count of at least 200 spermatozoa was performed per sample, using an optical microscope with immersion objective (100X), the vitality was evaluated by supravital eosin staining (0.7%)-nigrosin(1%), the results indicate that it does not exist a detrimental effect of the seminal plasma on the endosmotic response, being, on the contrary, superior in the whole semen; the vitality of the spermatozoa with and without seminal plasma is similar, however it decreases when it is reconstituted with seminal plasma, possibly due to the seminal plasma of another animal; there is no positive correlation between endosmosis and vitality, indicating that the latter would not necessarily reflect the integrity of the membrane, which is why it is recommended to perform this test routinely in alpaca semen exams.
Use of Androcoll-ETM to Separate Frozen-Thawed Llama Sperm From Seminal Plasma and Diluent
(Universidad Austral de Chile, 2021-01-21) Guillén Palomino, Crissthel Yverlin; Fumuso, Fernanda Gabriela; Bertuzzi, Mariana Lucía; Giuliano, Susana María; Velásquez González, Nicolás; Bariani, María Victoria
It is not easy to separate frozen-thawed South American camelid sperm from seminal plasma (SP) and diluents to be used for in vitro embryo production. The objective of this study was to evaluate Androcoll-E™ (AE) efficiency to separate llama sperm from SP and freezing extender in frozen-thawed semen. A total of 22 ejaculates from five Lama glama males were collected using electroejaculation. After performing semen analysis (sperm motility, concentration, viability, membrane function, and acrosome integrity), samples were cryopreserved with a diluent containing lactose, ethylenediaminetetraacetic acid (EDTA), egg yolk, and 7% dimethylformamide. After thawing, samples were divided in aliquots, one of which was used as a control and the others processed by AE. Experiment 1 (12 ejaculates): 100 μl of frozen-thawed semen was placed on top of 1,000 μl AE column and centrifuged at 800 g for 10 min. Experiment 2 (10 ejaculates): two samples of 100 μl of frozen-thawed semen were placed on two columns of 500 μl AE each, and both were centrifuged at 800 g for 10 and 20 min, respectively. Pellets were resuspended in Tyrode's albumin lactate pyruvate (TALP) medium, and sperm parameters were evaluated. A significant decrease in all sperm parameters was observed in thawed samples compared to raw semen. AE allowed the separation of frozen-thawed sperm from SP and freezing extender independently from the height of the column used and time of centrifugation assayed. Although no significant differences were found between AE columns, higher sperm recovery was observed with 500 μl of AE coupled with 20 min of centrifugation. Despite the significant decrease observed in sperm motility in AE samples, no changes in sperm viability, membrane function, and acrosome integrity were observed when comparing control thawed semen with the sperm recovered after AE (p > 0.05). The use of AE columns, either 500 or 1,000 μl, allows the separation of frozen-thawed llama sperm from SP and freezing extender, preserving the viability, membrane function, and acrosome integrity. Of the protocols studied, 800 g centrifugation during 20 min using a 500 μl column of AE would be the method of choice to process frozen-thawed llama semen destined for reproductive biotechnologies.