158 First Llama Born by in Vitro Fertilization

Abstract

The aim was to transfer alpaca and llama embryos obtained by IVF into recipient llamas and evaluate pregnancy and birth rates. Gametes were collected from samples of ovaries and testes collected from a local abattoir. Cumulus-oocyte complexes (COC) were recovered by aspiration of ovarian follicles using a 5-mL syringe, where oocytes with at least 3 layers of cumulus cells and a homogenous cytoplasm were matured in TCM-199 supplemented with 10% FCS, FSH (0.1 IU), and oestradiol 17β for 30 and 36 h with 5% CO2 in air, for alpaca and llama, respectively. After the maturation time, COC were placed in FERT-TALP solution 30 min before in vitro insemination with epididymal sperm of alpaca and llama as appropriate, the sperm were selected by swim-up method with centrifuging twice in 2 mL of SPERM-TALP supplemented with 6 mg mL−1 of fatty-free BSA, in an incubator with 5% CO2 in air set at 39°C for 45 min, oocytes were co-incubated with sperm concentration of 3 × 106 mL−1 for 18 to 20 h at 39°C with 5% CO2 in air. The IVF was carried out the day of ovulation induction of recipients. A group of 15 morphologically intact Day-8 blastocysts derived from in vitro culture with SOF serum were transferred nonsurgically into the uterine horn ipsilateral to the corpus luteum of 15 synchronized recipient llamas with an intravaginal device (controlled internal drug release) for 8 days. Then, 6 days after controlled internal drug release removal, ovulation was induced in recipients with the application of 1 mL of GnRH with previous ultrasound confirmation of the presence of a dominant follicle greater than 7 mm in diameter. Nine alpaca blastocysts and 6 llama blastocysts were transferred. The pregnancy rate was assessed by ultrasound at 45 days after transfer. The results obtained were as follows: for pregnancy rate, 33.3% (3/9) and 50% (3/6) for alpaca and llama embryos, respectively; for birth rate, 0.0% (0/9) and 16.7% (1/6) for alpaca and llama embryos, respectively. An alpaca fetus and 2 llama fetuses were aborted between 7 and 10 months of pregnancy, and only a llama gestation ended successfully, producing the first birth of the world of a llama bred by IVF in South American camelids, demonstrating that it is possible to obtain viable offspring in these species using this biotechnology.

PRODUCCIÓN Y TRANSFERENCIA DE EMBRIONES in vitro EN CAMÉLIDOS SUDAMERICANOS: NUEVAS OPORTUNIDADES Y DESAFIOS

“…45 días después se comprobó ecográficamente que se obtuvo 1 alpaca (33%) y 3 llamas (60%) preñadas. Estos resultados nos incentivaron a continuar con la transferencia de embriones (Landeo et al, 2016a), entonces realizamos 15 transferencias de embriones producidos por FIV, en esta oportunidad todas las receptoras fueron llamas. Se transfirieron 9 embriones de alpacas y 6 embriones de llamas producidos por FIV, se obtuvieron tasas de preñez de 33.3% (3/9) y 50% (3/6) en llamas transferidas con embriones de alpaca y llama respectivamente; y tasa de nacimiento de 0.0% (0/9) y 16.7% (1/6) para embriones de alpaca y llama respectivamente.…”

“…Existen desde hace varios años, trabajos de investigación en la producción de embriones por FIV en camélidos sudamericanos (Del Campo et al, 1994;Conde et al, 2008;Gamarra et al, 2008;Mellisho et al, 2014;. Asimismo, se han reportado gestaciones tempranas en llamas (Mendoza et al, 2013;Trasorras et al, 2014) y alpacas (Mendoza et al, 2013), y el nacimiento de la primera llama en el mundo (Landeo et al, 2016a) con la aplicación de esta biotecnología. Sin embargo, son necesarios mayores estudios para mejorar las tasas de preñez y de natalidad que permitan a mediano y largo plazo introducir esta biotecnología en los sistemas de producción de alpacas y llamas.…”

Production, Preservation, and Transfer of South American Camelid Embryos

“…Furthermore, the first report of a pregnancy in llama was by Trasorras et al ( 15 ) after the intrauterine transfer of embryos produced by IVF using fresh semen processed through Androcoll-E™ and oocytes obtained from superstimulated females (Figure 1 ). So far, the birth of only one llama (1/6) with IVF has been reported using slaughterhouse gametes ( 44 ). However, to date, the results obtained regarding early embryo development are poor, mainly because the quality of the blastocysts produced in vitro is not as good as that of blastocysts produced in vivo .…”

Comparison of superstimulatory protocols with different doses of eCG on ovarian response and oocyte recovery by follicular aspiration in llamas (Lama glama)

“…In South American camelids, reports of using IVP are limited. In these species, the main method reported for the obtention of cumulus-oocyte complexes (COCs) has been the dissection of ovaries from abattoir ( Ratto et al, 2005 ; Landeo et al, 2017 ). The use of ultrasound-guided transvaginal follicle aspiration to perform the ovum pick up (OPU) to collect COCs was reported with limited success in llamas ( Brogliatti et al, 2000 , Berland et al, 2011 ).…”