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

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).

1. Cerón-Mu-oz M, Gómez-Arroyave F, Ramírez-Toro J, Cifuentes T y Gutiérrez-Molina S. Parámetros genéticos para la producción de leche, grasa y proteína en búfalos de Colombia. Livestock Research for Rural Development. 2012 24(2), Article #30 URL Disponible en: http://www.lrrd.org/lrrd24/2/cero24030.htm
3. Perera B. Reproductive cycles of buffalo. Anim Reprod Sci 2011; 124(3):194-9. http://dx.doi.org/10.1016/j.anireprosci.2010.08.022
5. Berdugo Gutiérrez J. Historia de la aplicación de biotecnologías reproductivas en la cría del búfalo en Colombia. Uni-pluri/versidad 2013;12(3):87-91. URL Disponible en: http://aprendeenlinea.udea.edu.co/revistas/index.php/unip/article/viewFile/15160/13204
7. Drost M. Advanced reproductive technology in the water buffalo. Theriogenology 2007;68(3):450-3.
8. http://dx.doi.org/10.1016/j.theriogenology.2007.04.013
10. Boni R, Roviello S, Gasparrini B, Langella M, Zicarelli L. In vitro production of buffalo embryos in chemically defined medium. Buffalo J 1999;15:115-20.
12. Galli C, Duchi R, Crotti G, Lazzari G. Embryo production by ovum pick up in water buffalo. Theriogenology 1998; 49(1):400.
13. http://dx.doi.org/10.1016/S0093-691X(98)90753-1
15. Kumar A, Solanki V, Jindal S, Tripathi V, Jain G. Oocyte retrieval and histological studies of follicular population in buffalo ovaries. Anim Reprod Sci 1997; 47(3):189-95.
16. http://dx.doi.org/10.1016/S0378-4320(96)01588-6
18. Baruselli P, Mucciolo R, Visintin J, Viana W, Arruda R, Madureira E, et al. Ovarian follicular dynamics during the estrous cycle in buffalo (Bubalus bubalis). Theriogenology 1997; 47(8):1531-47. http://dx.doi.org/10.1016/S0093-691X(97)00159-3
20. Aspilcueta-Borquis RR, Sesana RC, Berrocal MHM, Seno LdO, Bignardi AB, El Faro L, et al. Genetic parameters for milk, fat and protein yields in Murrah buffaloes (Bubalus bubalis Artiodactyla, Bovidae). Genet Mol Biol 2010; 33(1):71-7.
21. http://dx.doi.org/10.1590/S1415-47572010005000005
23. Galeazzi P, Mercadante M, Silva J, de Albuquerque L, de Camargo G, Tonhati H. Analysis of culling probability in dairy buffalo using survival models. Animal. 2010; 4(8):1325-9.
24. http://dx.doi.org/10.1017/S1751731110000406
26. Hernández C, Casta-eda S, Lopez C, Arias C, Watanabe Y, Berdugo J. Aspiración folicular guiada por ultrasonido (OPU) y producción de embriones In vitro en búfalas de agua (Bubalus bubalis) en el trópico bajo colombiano. Rev Colomb Cienc Pecu 2009; 22(3):567.
28. Galli C, Berdugo J, Pacheco L, Angel A, Posada I, Lazzari G, et al. First pregnancies obtained after transfer frozen-thawed buffaline embryos produced In vitro in Colombia. Reprod Fertil Dev 2011; 24:190. http://dx.doi.org/10.1071/RDv24n1Ab157
30. Di Francesco S, Novoa MVS, Vecchio D, Neglia G, Boccia L, Campanile G, et al. Ovum pick-up and In vitro embryo production (OPU-IVEP) in Mediterranean Italian buffalo performed in different seasons. Theriogenology 2012; 77(1):148-54.
31. http://dx.doi.org/10.1016/j.theriogenology.2011.07.028
33. Sharma GT, Dubey PK, Nath A, Saikumar G. Co-culture of buffalo (Bubalus bubalis) preantral follicles with antral follicles: a comparative study of developmental competence of oocytes derived from in vivo developed and In vitro cultured antral follicles. Zygote. 2013; 21(3):286-94.
34. http://dx.doi.org/10.1017/S0967199411000700
36. Parrish JJ, Susko-Parrish JL, Leibfried-Rutledge ML, Critser ES, Eyestone WH, First NL. Bovine In vitro fertilization with frozen-thawed semen. Theriogenology 1986; 25:591-600.
37. http://dx.doi.org/10.1016/0093-691X(86)90143-3
39. Mondadori RG, Santin TR, Fidelis AAG, da Silva JS, Rumpf R, Báo SN. Ultrastructure of In vitro oocyte maturation in buffalo (Bubalus bubalis). Zygote 2010; 18(4):309-14.
40. http://dx.doi.org/10.1017/S0967199409990335
42. Sá Filho M, Carvalho N, Gimenes L, Torres-Júnior J, Nasser L, Tonhati H, et al. Effect of recombinant bovine somatotropin (bST) on follicular population and on In vitro buffalo embryo production. Anim Reprod Sci 2009; 113(1):51-9.
43. http://dx.doi.org/10.1016/j.anireprosci.2008.06.008
45. Manjunatha B, Ravindra J, Gupta P, Devaraj M, Nandi S. Oocyte recovery by ovum pick up and embryo production in river buffaloes (Bubalus bubalis). Reprod Domest Animal 2008; 43(4):477-80. http://dx.doi.org/10.1111/j.1439-0531.2007.00939.x
47. Neglia G, Gasparrini B, Caracciolo di Brienza V, Di Palo R, Campanile G, Antonio Presicce G, et al. Bovine and buffalo In vitro embryo production using oocytes derived from abattoir ovaries or collected by transvaginal follicle aspiration. Theriogenology 2003; 59(5):1123-30. http://dx.doi.org/10.1016/S0093-691X(02)01170-6
49. Gasparrini B, Sayoud H, Neglia G, Matos DGd, Donnay I, Zicarelli L. Glutathione synthesis during In vitro maturation of buffalo (Bubalus bubalis) oocytes: effects of cysteamine on embryo development. Theriogenology 2003; 60(5):943-52.
50. http://dx.doi.org/10.1016/S0093-691X(03)00098-0
52. Lu Y, Liang X, Zhang M, Wang W, Kitiyanant Y, Lu S, et al. Birth of twins after In vitro fertilization with flow-cytometric sorted buffalo (S) sperm. Anim Reprod Sci 2007; 100(1):192-6.
53. http://dx.doi.org/10.1016/j.anireprosci.2006.09.019
55. Galli C, Duchi R, Colleoni S, Lagutina I, Lazzari G. Ovum pick up, intracytoplasmic sperm injection and somatic cell nuclear transfer in cattle, buffalo and horses: from the research laboratory to clinical practice. Theriogenology. 2014;81(1):138-51.
56. http://dx.doi.org/10.1016/j.theriogenology.2013.09.008
58. Bó GA, Baruselli PS, Chesta PM, Martins CM. The timing of ovulation and insemination schedules in superstimulated cattle. Theriogenology 2006; 65(1):89-101.
59. http://dx.doi.org/10.1016/j.theriogenology.2005.10.008