Comparación de sistemas de cultivo de embriones de alpacas

Durand, Manuel Guido Pérez; Aragón, Juan Pompeyo Zevallos; Guerra, Uri Harold Perez

doi:10.18271/ria.2017.274

Cited by 3 publications

(2 citation statements)

References 17 publications

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“…Therefore, plant-based extenders are as effective at protecting spermatozoa during the cryopreservation process as egg yolk extenders. Another alpaca study showed that the in vitro fertilization rates were similar to those obtained in the present study, with the difference that non-cryopreserved spermatozoa were used [52]. However, there were studies in alpacas where higher fertilization rates have been observed compared to those reported in the present study [23].…”

Section: Discussionsupporting

confidence: 85%

Effect of Three Semen Extenders on Sperm Quality and In Vitro Fertilization Rates of Fresh and Cryopreserved Sperm Collected from Llama (Lama glama) Vas Deferens

Pérez-Durand,

Bustamante,

Machaca

et al. 2024

Animals

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The advances in Assisted Reproductive Technologies (ARTs) applied in South American camelid species are still scarce. The aim of this study was to compare the effects of three semen extenders, before and after the cryopreservation of spermatozoa obtained from the vas deferens, on sperm quality parameters and in vitro fertilization rates of llama (Lama glama) oocytes. Mature fertile llama males (Lama glama; n = 6; age: 48–60 mo.; BCS: ~2.7) were included in the study. Sperm samples were collected from each male using the surgical technique of the vas deferens deviation. Then, the sperm samples were pooled and diluted with the Tris-EY, Andromed®, or BioxCell® extender in order to subsequently carry out the sperm cryopreservation process. The sperm quality assessment related to each extender was performed before and after cryopreservation with regard to sperm morphological abnormalities, acrosome integrity, sperm viability, membrane permeability, and sperm motility traits. Moreover, in vitro fertilization (IVF) procedures were carried out to evaluate the in vitro fertility of the cryopreserved sperm samples using each extender. Overall, significant differences were observed before and after cryopreservation regarding acrosome integrity, sperm viability, membrane permeability, and sperm motility traits among the extenders used, where Tris-EY and Andromed® were better than BioxCell® (p < 0.05); however, no differences were observed regarding the sperm morphological abnormalities among extenders (p > 0.05). Moreover, multiple differences were observed with regard to the velocity and linearity kinematic parameters obtained by computerized analysis before and after the cryopreservation process, irrespective of the extender used (p < 0.05). Finally, differences were observed regarding the in vitro fertilization rates among the different extender-derived samples (p < 0.05). In conclusion, the sperm quality using Tris-EY and Andromed® was better before and after cryopreservation compared to that using BioxCell®. Although the number of fertilized oocytes obtained after the IVF process between Tris-EY and Andromed® was similar, Andromed®-derived samples showed the best sperm quality results before and after cryopreservation. This indicates that the cryopreservation extender is a determining factor in significantly improving in vitro fertilization rates when using sperm samples obtained from vas deferens in llama (Lama glama) males.

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Section: Discussionsupporting

confidence: 85%

Effect of Three Semen Extenders on Sperm Quality and In Vitro Fertilization Rates of Fresh and Cryopreserved Sperm Collected from Llama (Lama glama) Vas Deferens

Pérez-Durand,

Bustamante,

Machaca

et al. 2024

Animals

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“…Furthermore, ejaculated sperm of these species have a particular motility pattern, they possess oscillatory motility with practically no progressive motility (2)(3)(4). Due to these characteristics, several in vitro SAC embryo production studies have reported the use of sperm recovered from animals with deviated deferent ducts or collected from the epididymis after death or castration, since those samples are free of seminal plasma (SP) and have sperm with progressive motility (5)(6)(7)(8)(9)(10)(11)(12)(13). Moreover, the absence of high viscosity and thread formation are advantages when processing these SP-free samples.…”

Section: Introductionmentioning

confidence: 99%

Use of Androcoll-ETM to Separate Frozen-Thawed Llama Sperm From Seminal Plasma and Diluent

et al. 2021

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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.

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Efecto de factores embriotróficos a diferentes tensiones de oxígeno en cultivo in vitro sobre el desarrollo embrionario de alpacas hasta la etapa de blastocisto

Mamani¹

2022

lgr

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La alpaca es el camélido sudamericano doméstico de mayor importancia para Perú, país que cuenta con el 87% de la población a nivel regional. Las formas tradicionales de reproducción, no garantizan la calidad genética de los animales, por lo que la reproducción in vitro es una alternativa para su mejoramiento. El objetivo de la investigación fue analizar cómo influyen factores embriotróficos y tensiones de oxígeno en el desarrollo in vitro de ovocitos de alpaca hasta etapa de blastocistos. Se recolectaron ovarios de animales sacrificados y los ovocitos madurados en medio TCM-199, piruvato de sodio, glutamina, estradiol, hormona folículo estimulante, hormona luteinizante, factor de crecimiento epidermal, suero fetal bovino y gentamicina, por 32 horas a 38.5 ºC, al 5% CO2 y humedad relativa mayor de 95%, se fecundaron con semen fresco y cultivado en medio KSOMaa durante 48 h. Se utilizaron dos factores embriotróficos (EGF y IGF-I), dos tensiones de O2 (5 y 20%), más un grupo control sin factor embriotrófico y tres variables respuesta (ovocitos, divisiones y blastocistos). Se aplicó la técnica estadística de análisis de conglomerados para establecer diferencias entre los tratamientos con ? = 0,05. Se observaron diferencias significativas para cada variable respuesta, con máxima producción de divisiones de 24,88% para EGF a 6% O2 y blastocistos 18,4% para IGF-I a 6% O2. Se concluye que la adición de los factores embriotróficos a los medios de cultivo embrionario in vitro y una baja tensión de oxígeno 6% son favorables para el desarrollo embrionario en alpacas.

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Comparación de sistemas de cultivo de embriones de alpacas

Cited by 3 publications

References 17 publications

Effect of Three Semen Extenders on Sperm Quality and In Vitro Fertilization Rates of Fresh and Cryopreserved Sperm Collected from Llama (Lama glama) Vas Deferens

Effect of Three Semen Extenders on Sperm Quality and In Vitro Fertilization Rates of Fresh and Cryopreserved Sperm Collected from Llama (Lama glama) Vas Deferens

Use of Androcoll-ETM to Separate Frozen-Thawed Llama Sperm From Seminal Plasma and Diluent

Efecto de factores embriotróficos a diferentes tensiones de oxígeno en cultivo in vitro sobre el desarrollo embrionario de alpacas hasta la etapa de blastocisto

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