Establishing an in vitro production program for buffalo embryos (Bubalus bubalis) in Colombia

Objective. Evaluate the results of the standardization of the in vitro production program of buffalo embryos, using oocytes obtained by ultrasound guided oocyte puncture during the 2012 breeding season in Colombia. Materials and methods. Fifty seven buffalo females were selected for ultrasound guided transvaginal aspiration of follicles, oocytes were identified within follicular fluid, classified and transported to the laboratory and matured in vitro for 18 to 20 hours. Frozen semen of seven Mediterranean bulls were used, motile sperm was obtained using the Percoll technique and oocytes were inseminated with 2 million sperm/ml. Presumptive zygotes were cultured for 6 days, grade 1 embryos obtained were frozen using ethylene glycol. Embryos were transferred to females on day 5 during natural cycle. Results. 97 aspirations were performed on the 57 animals, in 8.2% of the aspirations no oocytes were found. 8 oocytes/aspiration were obtained. Of the 783 oocytes, 92% were classified as viable (721/783) and were fertilized. The cleavage and blastocyst rate were 23% and 19% respectively. 37 embryos were transferred and 11 pregnancies were obtained, confirmed by rectal palpation 60 days after transfer, with a pregnancy rate of 29.7%. Conclusions. The results presented here are comparable with others in literature and shows the feasibility of producing in vitro embryos and pregnancies after the standardization of current protocols, with normal and sexed semen and transfer during natural cycle in buffalo.Key words: Buffaloes, embryo, fertilization in vitro, reproduction (Source: MeSH)

Establishing an in vitro production program for buffalo embryos (Bubalus bubalis) in Colombia

Objective. Evaluate the results of the standardization of the in vitro production program of buffalo embryos, using oocytes obtained by ultrasound guided oocyte puncture during the 2012 breeding season in Colombia. Materials and methods. Fifty seven buffalo females were selected for ultrasound guided transvaginal aspiration of follicles, oocytes were identified within follicular fluid, classified and transported to the laboratory and matured in vitro for 18 to 20 hours. Frozen semen of seven Mediterranean bulls were used, motile sperm was obtained using the Percoll technique and oocytes were inseminated with 2 million sperm/ml. Presumptive zygotes were cultured for 6 days, grade 1 embryos obtained were frozen using ethylene glycol. Embryos were transferred to females on day 5 during natural cycle. Results. 97 aspirations were performed on the 57 animals, in 8.2% of the aspirations no oocytes were found. 8 oocytes/aspiration were obtained. Of the 783 oocytes, 92% were classified as viable (721/783) and were fertilized. The cleavage and blastocyst rate were 23% and 19% respectively. 37 embryos were transferred and 11 pregnancies were obtained, confirmed by rectal palpation 60 days after transfer, with a pregnancy rate of 29.7%. Conclusions. The results presented here are comparable with others in literature and shows the feasibility of producing in vitro embryos and pregnancies after the standardization of current protocols, with normal and sexed semen and transfer during natural cycle in buffalo.Objetivo. Evaluar los resultados de la estandarización de la técnica de producción in vitro de embriones de búfalo, a partir de oocitos obtenidos por punción folicular durante la estación reproductiva del 2012 en una hacienda en Cordoba, Colombia. Materiales y métodos. Cincuenta y siete búfalas fueron seleccionadas para aspiración transvaginal de folículos guiada por ultrasonido, los oocitos fueron identificados y madurados in vitro. Se utilizó semen congelado de 7 búfalos de la raza Mediterráneo para la fertilización in vitro. La fracción móvil fue separada en un gradiente de Percoll, los oocitos fueron inseminados con 2 millones de espermatozides/mL, los presuntos cigotos fueron cultivados por 6 días y los embriones grado 1 obtenidos fueron congelados utilizando etilenglicol. Posteriormente, los embriones fueron transferidos el día 5 post estro en búfalas en ciclo natural. Resultados. En las 57 búfalas, se realizaron 97 aspiraciones foliculares, no se obtuvieron oocitos en el 8.2% de los procedimientos. Se obtuvieron en promedio 8 oocitos por búfala en cada sesión de aspiración. Se recolectaron 783 oocitos de los cuales fueron clasificados como viables el 92% (721/783), se obtuvo una taza de clivaje del 23% y de blastocistos del 19%. De 37 embriones transferidos se obtuvieron 11 preñeces, confirmadas por palpación rectal a los 60 días postransferencia, obteniéndose una taza de preñez del 29.7% Conclusiones. Los resultados presentados en este trabajo son comparables con los de la literatura, en la cual se muestra cómo es posible obtener embriones de semen convencional y sexado, además de producir gestaciones con protocolos estándar de fertilización in vitro y transferencia en ciclos con celo natural adaptados para la especie