RESUMO

This study assessed the outcomes of nonsurgical embryo recovery (NSER) after superovulation (SOV) in five locally adapted Brazilian breeds of sheep and goats. The objective was to evaluate the feasibility and efficiency of using SOV combined with a less-invasive embryo collection technique for supplying the Brazilian animal gene bank with germplasm from specific genotypes of interest. Morada Nova (n = 20), Santa Inês (n = 20), and Somalis (n = 20) ewes received an intravaginal progesterone (330 mg) device for 9 days, while Canindé (n = 15) and Moxotó (n = 15) goats received an intravaginal medroxyprogesterone acetate (60 mg) device for 6 days. All females received 133 mg of porcine follicle-stimulating hormone (pFSH) administrated in six decreasing doses 12 hours apart, starting 60 hours before device removal, plus 37.5 µg of d-cloprostenol at the fifth and sixth pFSH dose. Donors in estrus were mated with fertile males. The corpora lutea (CL) number was assessed by ultrasonography 1 day before NSER. On day 6.5 or 7 after estrus, NSER was performed following hormonally induced cervical relaxation. A total of 97% of sheep and 90% of goats responded with estrus, and among those, 91% of sheep and 85% of goats presented a CL. In ewes, the numbers of CL were greater (p < 0.05) in the Santa Inês breed, while similar (p > 0.05) CL numbers were found among the goat breeds. All viable embryos were freezable (excellent and good quality) and the number per donor was 7.8 for sheep and 4.9 for goats. All parameters of NSER efficiency, embryo yield, and fertility post-NSER did not differ (p > 0.05) between breeds among each species. The SOV-NSER procedures applied for an embryo biobank supply of locally adapted Brazilian breeds of small ruminants were efficient regarding production of cryopreservable embryos, and preservation of donor fertility. Therefore, SOV followed by NSER is recommended for embryo biobank assembly in sheep and goats.

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RESUMO

Superovulatory response and embryo yield in 19 Morada Nova and 20 Somalis Brasileira ewes was analyzed. All animals were synchronized with the insertion of an intravaginal device (CIDR®) on Day 0, replaced by a new device on Day 7, which remained in place until Day 14 and superovulated with 133mg of porcine FSH (pFSH) in decreasing doses at 12h intervals from Day 12 until Day 15 of the treatment, and a single dose of equine chorionic gonadotropin (eCG, 200UI) on Day 14 (i.e., administered in CIDR removal). Fifty hours after CIDR® removal, females were inseminated by laparoscopy. All embryos were recovered by laparotomy 5 days after insemination. Sheep which responded to the superovulation protocol (P>0.05) included 74% of the Morada Nova ewes and 50% of the Somalis Brasileira ewes. Morada Nova showed better results (P<0.05) than Somalis Brasileira in number of ovulations (15.38 ± 5.24 vs. 10.56 ± 2.83), total structures (11.00 ± 7.55 vs. 3.33 ± 1.94) and embryo yields (6.79 ± 5.35 vs. 2.90 ± 2.18). Despite the high fertilization rate, degenerate embryo rate was high for both breeds, with an overall rate of 39% (57/145). In conclusion, superovulatory response and embryo yields in Morada Nova ewes were considered sufficient to justify the use of this procedure in genetic resources conservation programs. However, improvements to embryo quality and control of precocious regression of corpus luteum are necessary to produce better results in the MOET program, with minimal variations and maximum embryo yield in Morada Nova and Somalis Brasileira ewes.(AU)

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RESUMO

Superovulatory response and embryo yield in 19 Morada Nova and 20 Somalis Brasileira ewes was analyzed. All animals were synchronized with the insertion of an intravaginal device (CIDR®) on Day 0, replaced by a new device on Day 7, which remained in place until Day 14 and superovulated with 133mg of porcine FSH (pFSH) in decreasing doses at 12h intervals from Day 12 until Day 15 of the treatment, and a single dose of equine chorionic gonadotropin (eCG, 200UI) on Day 14 (i.e., administered in CIDR removal). Fifty hours after CIDR® removal, females were inseminated by laparoscopy. All embryos were recovered by laparotomy 5 days after insemination. Sheep which responded to the superovulation protocol (P>0.05) included 74% of the Morada Nova ewes and 50% of the Somalis Brasileira ewes. Morada Nova showed better results (P<0.05) than Somalis Brasileira in number of ovulations (15.38 ± 5.24 vs. 10.56 ± 2.83), total structures (11.00 ± 7.55 vs. 3.33 ± 1.94) and embryo yields (6.79 ± 5.35 vs. 2.90 ± 2.18). Despite the high fertilization rate, degenerate embryo rate was high for both breeds, with an overall rate of 39% (57/145). In conclusion, superovulatory response and embryo yields in Morada Nova ewes were considered sufficient to justify the use of this procedure in genetic resources conservation programs. However, improvements to embryo quality and control of precocious regression of corpus luteum are necessary to produce better results in the MOET program, with minimal variations and maximum embryo yield in Morada Nova and Somalis Brasileira ewes.(AU)

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RESUMO

El mantenimiento en campo de los Bancos de Germoplasma resulta muy costoso, además de los riesgos a que se exponen. El cultivo de tejidos constituye una solución a estos problemas siendo conveniente utilizar una combinación de técnicas de almacenamiento en los cultivos de propagación vegetativa para no depender de una sola. El cultivo in vitro ofrece nuevas alternativas para el mejoramiento de la productividad y la producción de material de siembra sano en malanga (Xanthosoma spp.). La presente investigación se desarrolló en el Laboratorio de Cultivo de Tejidos del Instituto de Investigaciones de Viandas Tropicales (INIVIT), Cuba, con el objetivo de estudiar las condiciones para la conservación en crecimiento mínimo in vitro de germoplasma de esta especie. Como material vegetal se utilizó el clon de Malanga Xanthosoma "INIVIT MX-2008". El establecimiento del material vegetal y su posterior multiplicación fueron realizadas según la metodología recomendada por García et al. (1999). Para la conservación en medio de cultivo de crecimiento mínimo se utilizó el medio basal MS y se estudiaron 15 tratamientos que combinaron concentraciones de Manitol (regulador osmótico) (1,5; 3 y 4%) y Nitrato de plata (inhibidor de la acción etileno) (0, 2, 4, 8, 10 mg.L-1). Se concluye que es posible conservar in vitro los recursos genéticos de malanga Xanthosoma durante más de 10 meses, en un medio de cultivo compuesto por sales y vitaminas MS suplementado con 4% de manitol y 4 mg.L-1 de Nitrato de plata. Las plantas propagadas a partir de este medio de cultivo se recuperaron exitosamente. La mayor concentración de manitol en el medio de cultivo pudo haber influido en la mejor recuperación del material conservado.

Maintenance field genebanks are costly, in addition to the risks they face; to that effect on tissue culture is a solution to these problems. In vegetative propagated crops is desirable to use a combination of storage technology rather than relying on just one. in vitro culture provides an alternative for improving productivity and production of healthy planting material of taro (Xanthosoma spp.). This research was conducted in the Tissue Culture Laboratory of the Research Institute of Tropical Crops. Our objective was study the conditions for minimal growth conservation in vitro germplasm in this species. As plant material was used clone of Taro Xanthosoma 'INIVIT MX-2008'. The establishment of the plant material and its subsequent multiplication were carried out according to the methodology recommended by García et al. (1999). For the maintenance in culture of minimal growth basal medium MS was used and studied 15 treatments with combined concentrations of mannitol (osmotic regulator) (1.5, 3 and 4%) and silver nitrate (Ethylene inhibitor) (0, 2, 4, 8, 10 mg.L-1). It concludes that it is possible to conserve taro Xanthosoma genetic resources in vitro, for over 10 months in a culture medium composed of MS salts and vitamins and supplemented with 4% mannitol and 4 mg.L-1 of silver nitrate. Plants propagated from this culture medium were recovered successfully. The presence of higher concentrations of mannitol, may have influenced that increases survival of preserved material. O3.

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