In vitro embryo production in Nelore cows supplemented with calcium salts of long-chain fatty acid

Moacir Rogério de Souza, Luciano Soares de Lima, Fabio Luiz Bim Cavalieri, Luiz Paulo Rigolon, Wallacy Barbacena Rosa dos Santos, Josmar Mazucheli, Alexandre Leseur dos Santos, Geraldo Tadeu dos Santos


This study was performed to evaluate the effects of supplementation of lactating Nelore cows with calcium salts of long-chain fatty acids (Ca-LCFA) of soybean oil on in vitro oocytes competency, cleavage rate, embryo production rate, pregnancy rate, and sex ratio of transferred embryos. Fifteen multiparous Nelore cows grazed on a Tifton 85 pasture were allotted to two treatments: (1) supplement without fat inclusion (CONT, n = 6) and (2) supplement containing 47.2 g Ca-LCFA /kg (to provide an intake of 0.2 kg Ca-LCFA/day) based on dry matter (n = 9). Animals were subjected to 2.53 ± 1.35 rounds of consecutive ovum pick up at intervals of 27.65 ± 21.72 days. Oocytes were morphologically evaluated, selected, maturated, fertilized, and cultivated for embryo production in vitro and then transferred into recipients who had previously undergone an embryo transfer protocol at a fixed time. The number of aspirated oocytes, the proportion of viable oocytes, cleavage rate, embryo percentage, pregnancy rate, and sex ratio were not affected when Nelore cows were supplemented with Ca-LCFA of soybean oil compared with non-supplemented Nelore cows. In conclusion, our results suggest that feeding a supplement containing 47.2 g Ca-LCFA/kg (to provide an intake of 0.2 kg Ca-LCFA/day) to Nelore cows does not affect in vitro embryo production in lactating Nelore cows. However, further studies using higher dietary doses of fat could be useful to determine the potential of fat supplementation to improve in vitro embryo production.


Beef cattle; Bos indicus; Fatty acids; Oocytes; Ovum pick up; Pregnancy.

Full Text:



Aardema, H., Vos, P. L. A. M., Lolicato, F., Roelen, B. A. J., Knijn, H. M., Vaandrager, A. B.,… Gadella, B. M. (2011). Oleic acid prevents detrimental effects of saturated fatty acids on bovine oocyte developmental competence1. Biology of Reproduction, 85(1), 62-69. doi: 10.1095/biolreprod.110.088815

Adamiak, S. J., Powell, K., Rooke, J. A., Webb, R., & Sinclair, K. D. (2006). Body composition, dietary carbohydrates and fatty acids determine post-fertilisation development of bovine oocytes in vitro. Reproduction, 131(2), 247-258. doi: 10.1530/rep.1.00871

Association of Official Analytical Chemists (1998). Official methods of analysis (16nd ed.). Gaithersburg, M.P.: AOAC.

Bilby, T. R., Block, J., Amaral, B. C. do, Sa, O., Fº., Silvestre, F. T., Hansen, P. J.,… Thatcher, W. W. (2006). Effects of dietary unsaturated fatty acids on oocyte quality and follicular development in lactating dairy cows in summer. Journal of Dairy Science, 89(10), 3891-3903. doi: 10.3168/jds.S0022-0302(06) 72432-8

Bó, G. A., Cedeño, A., & Mapletoft, R. J. (2019). Strategies to increment in vivo and in vitro embryo production and transfer in cattle. Animal Reproduction, 16(3), 411-422. doi: 10.21451/1984-3143-AR 2019-0042

Cerri, R. L. A., Juchem, S. O., Chebel, R. C., Rutigliano, H. M., Bruno, R. G. S., Galvão, K. N.,… Santos, J. E. P. (2009). Effect of fat source differing in fatty acid profile on metabolic parameters, fertilization, and embryo quality in high-producing dairy cows. Journal of Dairy Science, 92(4), 1520-1531. doi: 10.3168/ jds.2008-1614

Cooke, R. F. (2019). Early career achievement award: Supplementing omega-6 fatty acids to enhance early embryonic development and pregnancy establishment in Bos indicus and B. taurus beef cows. Journal of Animal Science, 97(1), 485-495. doi: 10.1093/jas/sky414

Curran, S., Kastelic, J. P., & Ginther, O. J. (1989). Determining sex of the bovine fetus by ultrasonic assessment of the relative location of the genital tubercle. Animal Production Science, 19(3), 217-227. doi: 10.1016/0378-4320(89)90095-X

Ferguson, J. D., Sklan, D., Chalupa, W. V., & Kronfeld, D. S. (1990). Effects of hard fats on in vitro and in vivo rumen fermentation, milk production, and reproduction in dairy cows. Journal of Dairy Science, 73(10), 2864-2879. doi: 10.3168/jds.S0022-0302(90)78974-6

Fouladi-Nashta, A. A., Gutierrez, C. G., Gong, J. G., Garnsworthy, P. C., & Webb, R. (2007). Impact of dietary fatty acids on oocyte quality and development in lactating dairy cows. Biology of Reproduction, 77(1), 9-17. doi: 10.1095/biolreprod.106.058578

Fouladi-Nashta, A. A., Wonnacott, K. E., Gutierrez, C. G., Gong, J. G., Sinclair, K. D., Garnsworthy, P. C., & Webb, R. (2009). Oocyte quality in lactating dairy cows fed on high levels of n-3 and n-6 fatty acids. Reproduction, 138(5), 771-781. doi: 10.1530/REP-08-0391

Funston, R. N. (2004). Fat supplementation and reproduction in beef females. Journal of Animal Science, 82(E-Suppl.) E154-161. doi: 10.2527/2004.8213_supplE154x

Green, M. P., Spate, L. D., Parks, T. E., Kimura, K., Murphy, C. N., Williams, J. E.,… Roberts, R. M. (2008). Nutritional skewing of conceptus sex in sheep: effects of a maternal diet enriched in rumen-protected polyunsaturated fatty acids (PUFA). Reproductive Biology and Endocrinology?: RB&E, 6(21), 1-11. doi: 10.1186/1477-7827-6-21

Gutiérrez-Adán, A., Behboodi, E., Andersen, G. B., Medrano, J. F., & Murray, J. D. (1996). Relationship between stage of development and sex of bovine IVM-IVF embryos cultured in vitro versus in the sheep oviduct. Theriogenology, 46(3), 515-528. doi: 10.1016/0093-691X(96)00173-2

Gutiérrez-Adán, A., Lonergan, P., Rizos, D., Ward, F. A., Boland, M. P., Pintado, B., & de la Fuente, J. (2001). Effect of the in vitro culture system on the kinetics of blastocyst development and sex ratio of bovine embryos. Theriogenology, 55(5), 1117-1126. doi: 10.1016/S0093-691X(01)00471-X

Hiller, B. (2014). Recent developments in lipid metabolism in ruminants - the role of fat in maintaining animal health and performance. Animal Production Science, 54(10), 1549-1560. doi: 10.1071/AN14555

Lonergan, P., & Fair, T. (2008). In vitro-produced bovine embryos: dealing with the warts. Theriogenology, 69(1), 17-22. doi: 10.1016/j.theriogenology.2007.09.007

Lopes, C. N., Cooke, R. F., Reis, M. M., Peres, R. F., & Vasconcelos, J. L. (2011). Strategic supplementation of calcium salts of polyunsaturated fatty acids to enhance reproductive performance of Bos indicus beef cows. Journal of Animal Science, 89(10), 3116-3124. doi: 10.2527/jas.2011-3909

Lopes, C. N., Scarpa, A. B., Cappellozza, B. I., Cooke, R. F., & Vasconcelos, J. L. M. (2009). Effects of rumen-protected polyunsaturated fatty acid supplementation on reproductive performance of Bos indicus beef cows. Journal of Animal Science, 87(12), 3935-3943. doi: 10.2527/jas.2009-2201

Mattos, R., Staples, C. R., & Thatcher, W. W. (2000). Effects of dietary fatty acids on reproduction in ruminants. Reviews of Reproduction, 5(1), 38-45. doi: 10.1530/ror.0.0050038

Meikle, A., Brun, V. de, Carriquiry, M., Soca, P., Sosa, C., Adrien, M. de L.,… Abecia, J. A. (2018). Influences of nutrition and metabolism on reproduction of the female ruminant. Animal Reproduction, 15(1), 899-911. doi: 10.21451/1984-3143-AR2018-0017

Mertens, D. R. (2002). Gravimetric determination of amylase-treated neutral detergent fiber in feeds with refluxing in beakers or crucibles: collaborative study. Journal of AOAC International, 85(6), 1217-1240. doi: 10.1093/jaoac/85.6.1217

National Research Council (1996). Nutrients requirements of beef cattle (7nd ed.). Washington, D.C.: The National Academies Press.

Parrish, J. J., Susko-Parrish, J. L., Leibfried-Rutledge, M. L., Critser, E. S., Eyestone, W. H., & First, N. L. 1986. Bovine in vitro fertilization with frozen-thawed semen. Theriogenology 25(4), 591-600. doi: 10. 1016/0093-691x(86)90143-3

Peterson, A. J., & Lee, R. S. (2003). Improving successful pregnancies after embryo transfer. Theriogenology, 59(2), 687-697. doi: 10.1016/s0093-691x(02)01248-7

Ponter, A. A., Guyader-Joly, C., Nuttinck, F., Grimard, B., & Humblot, P. (2012). Oocyte and embryo production and quality after OPU-IVF in dairy heifers given diets varying in their n-6/n-3 fatty acid ratio. Theriogenology, 78(3), 632-645. doi: 10.1016/j.theriogenology.2012.03.009

Pontes, J. H., Nonato, I., Jr., Sanches, B. V., Ereno, J. C., Jr., Uvo, S., Barreiros, T. R.,... Seneda, M. M. (2009). Comparison of embryo yield and pregnancy rate between in vivo and in vitro methods in the same Nelore (Bos indicus) donor cows. Theriogenology, 71(4), 690-697. doi: 10.1016/j.theriogenology.2008. 09.031

Richards, M. W., Spitzer, J. C., & Warner, M. B. (1986). Effect of varying levels of postpartum nutrition and body condition at calving on subsequent reproductive performance in beef cattle. Journal of Animal Science, 62(2), 300-306. doi: 10.2527/jas1986.622300x

Rizos, D., Clemente, M., Bermejo-Alvarez, P., de La Fuente, J., Lonergan, P., & Gutierrez-Adan, A. (2008). Consequences of in vitro culture conditions on embryo development and quality. Reproduction in Domestic Animals = Zuchthygiene, 43(Suppl. 4), 44-50. doi: 10.1111/j.1439-0531.2008.01230.x

Rizos, D., Ward, F., Duffy, P., Boland, M. P., & Lonergan, P. (2002). Consequences of bovine oocyte maturation, fertilization or early embryo development in vitro versus in vivo: implications for blastocyst yield and blastocyst quality. Molecular Reproduction and Development, 61(2), 234-248. doi: 10.1002/ mrd.1153

Robertson, I., & Nelson, R. E. (1998). Certification and identification of the embryo. In D. A. Stringfellow, & S. M. Seidel (Eds.), Manual of the international embryo transfer society (pp. 103-134). Savoy: IETS

Santos, J. E. P., Bilby, T. R., Thatcher, W. W., Staples, C. R., & Silvestre, F. T. (2008). Long chain fatty acids of diet as factors influencing reproduction in cattle. Reproduction in Domestic Animals, 43(Suppl. 2), 23-30. doi: 10.1111/j.1439-0531.2008.01139.x

Siqueira, L. G., Torres, C. A., Amorim, L. S., Souza, E. D., Camargo, L. S., Fernandes, C. A., & Viana, J. H. (2009). Interrelationships among morphology, echotexture, and function of the bovine corpus luteum during the estrous cycle. Animal Production Science, 115(1-4), 18-28. doi: 10.1016/j.anireprosci.2008. 11.009

Souza, M. R. de, Santos, G. T. dos, Lima, L. S. de, Mazucheli, J., Zeoula, L. M., Rigolon, L. P., & Cavalieri, F. L. B. (2016). Oocyte production in Nellore cows supplemented with long-chain fatty acid soaps. Revista Brasileira de Zootecnia, 45(3), 138-143. doi: 10.1590/S1806-92902016000300008

Swecker, W. S. (2014). Interaction of nutrition and reproduction in the beef cow. In R. M. Hopper, Bovine reproduction (pp. 276-282). Hoboken, NJ: John Wiley & Sons, Inc.

Thangavelu, G., Colazo, M. G., Ambrose, D. J., Oba, M., Okine, E. K., & Dyck, M. K. (2007). Diets enriched in unsaturated fatty acids enhance early embryonic development in lactating Holstein cows. Theriogenology, 68(7), 949-957. doi: 10.1016/j.theriogenology.2007.07.002

Van Wagtendonk-de Leeuw, A. M., Mullaart, E., de Roos, A. P., Merton, J. S., den Daas, J. H., Kemp, B., & de Ruigh, L. (2000). Effects of different reproduction techniques: AI MOET or IVP, on health and welfare of bovine offspring. Theriogenology, 53(2), 575-597. Recovered from http://www.ncbi.nlm. 35051

Zeron, Y., Sklan, D., & Arav, A. (2002). Effect of polyunsaturated fatty acid supplementation on biophysical parameters and chilling sensitivity of ewe oocytes. Molecular Reproduction and Development, 61(2), 271-278. doi: 10.1002/mrd.1156


Semina: Ciênc. Agrár.
Londrina - PR
E-ISSN 1679-0359
DOI: 10.5433/1679-0359
Este obra está licenciado com uma Licença Creative Commons Atribuição-NãoComercial 4.0 Internacional