The present study aims to evaluate the effect of two culture media on the production of in vitro embryos in alpacas (Vicugna pacos). The ovaries were transported at 10.52° C in 0.9% saline solution supplemented with gentamicin. The ovaries were transported at 10.52° C in 0.9% physiological saline solution supplemented with gentamicin. 492 ovaries were used throughout the experiment. 2142 oocytes of quality I, II and III were recovered. The oocytes were matured in vitro for 32 h and were subsequently fertilized (incubated for 18 h) with sperm obtained from the tail of the epididymis and selected with a 45/90 percoll gradient. Then, the presumed zygotes were denuded from the cumulus cells, to later be cultured in two culture media: synthetic oviductal fluid medium (SOFaa) and simple optimized potassium medium (KSOMaa) and incubated at 38.5 ° C, 5 % CO2, 5%, O2, and 90% relative humidity for 7 days. Morula and blastocyst rate evaluation was performed at the end of embryo culture. The morula rate at 7 days was 41.49 ± 10.52 and 41.51 ± 6.50% for KSOMaa and SOFaa, respectively (P <0.05). The blastocyst rate for the two culture media KSOMaa and SOFaa, was 14.08 ± 5.17 and 11.73 ± 5.69 %, respectively, and there were no statistical differences (P˃0.05). The embryonic quality in KSOMaa and SOFaa media did not show statistical differences. In conclusion, the KSOMaa and SOFaa culture medium can be used in the production of in vitro embryos of alpacas
El estudio tuvo como objetivo evaluar el efecto de la colagenasa, pipeteo e incubación para disminuir la filancia del semen y su relación con la calidad espermática en alpacas para su criopreservación. Las muestras de semen se obtuvieron por vagina artificial de machos de 4-7 años en el Laboratorio de Biotecnología Reproductiva de la Estación Experimental Agraria Canaán del INIA (Ayacucho, Perú). Se evaluó las características macroscópicas (volumen, pH y filancia) y microscópicas (motilidad, concentración y vitalidad) del semen. Cada muestra se distribuyó en tres alícuotas y diluida con dilutor comercial AndroMed® en proporción de 1:1 a 37 °C y distribuidas en tratamientos: T1=Colagenasa al 5%; T2=Pipeteo 10 veces y T3=Incubación a 37 °C durante 10 min. Las características microscópicas se evaluaron a los 0, 20, 40, 60 y 120 min pos-dilución. La filancia disminuyó significativamente de 0.95, 0.89 y 0.91 cm a 0.26, 0.27 y 0.33 cm a los 120 min pos-dilución para T1, T2 y T3, respectivamente (p<0.05), pero sin diferencias entre tratamientos. La motilidad espermática y la vitalidad no fueron afectadas por los tiempos pos-dilución, pero la motilidad disminuyó significativamente (p<0.05) luego de la dilución del semen. Se concluye que el uso de colagenasa, pipeteo e incubación disminuyeron la filancia del semen de alpaca pos-dilución, afectando la motilidad y vitalidad espermática.
The objective of this research was to evaluate the effect of the use of two sperm selection techniques for in vitro production of alpaca embryos. The ovaries and testis were collected from the local slaughterhouse and transport to 37 ° C in saline solution (0.9%) supplemented with gentamicin. Quality I, II and II oocytes were incubated in a maturation medium for 32 h at 38.5 ° C and 5% O2 and 5% CO2. For in vitro fertilization, sperm from the epididymis were selected using the Percoll gradient and Swim up technique. 18h after the oocytes were incubated with the sperm, these were denuded from the cumulus cells and cultured in SOFaa culture medium for 7 days. Morula and blastocyst rate and their morphological quality are evaluated at day 7 of culture. From a total of 370 ovaries, 1,137 oocytes were recovered, making an average of 3.6 oocytes / ovary. After the maturation and fertilization process and in vitro culture, the blastocyst rate was 8.43 ± 6.04% and 3.89 ± 1.75%, for oocytes fertilized with sperm selected with Percoll gradient and Swim up, respectively, not finding significant statistical differences (p> 0.05), between the groups. In conclusion, the in vitro fertilization of alpaca oocytes with spermatozoa selected with two selection techniques (percoll and swim up) did not significantly influence the quantity and quality of morulae and blastocysts at day 7 of embryo culture.
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