Citrus Pulp as a Dietary Source of Antioxidants for Lactating Holstein Cows Fed Highly Polyunsaturated Fatty Acid Diets

  • Santos, G.T. (Departamento de Zootecnia, Universidade Estadual de Maringa) ;
  • Lima, L.S. (Departamento de Zootecnia, Universidade Estadual de Maringa) ;
  • Schogor, A.L.B. (Departamento de Zootecnia, Universidade Estadual de Maringa) ;
  • Romero, J.V. (Departamento de Zootecnia, Universidade Estadual de Maringa) ;
  • De Marchi, F.E. (Departamento de Zootecnia, Universidade Estadual de Maringa) ;
  • Grande, P.A. (Departamento de Zootecnia, Universidade Estadual de Maringa) ;
  • Santos, N.W. (Departamento de Zootecnia, Universidade Estadual de Maringa) ;
  • Kazama, R. (Departamento de Zootecnia e Desenvolvimento Rural, Universidade Federal de Santa Catarina)
  • Received : 2013.12.20
  • Accepted : 2014.03.27
  • Published : 2014.08.01


The effects of feeding pelleted citrus pulp (PCP) as a natural antioxidant source on the performance and milk quality of dairy cows fed highly polyunsaturated fatty acid (FA) diets were evaluated. Four lactating Holstein cows were assigned to a $4{\times}4$ Latinsquare. Treatments, on a dry matter (DM) basis, were i) control diet; ii) 3% soybean oil; iii) 3% soybean oil and 9% PCP and; iv) 3% soybean oil and 18% PCP. When cows fed on citrus pulp, the DM intake tended to decrease. The total tract apparent digestibility of DM and ether extract decreased when cows fed on the control diet compared to other diets. Cows fed PCP had higher polyphenols and flavonoids content and higher total ferric reducing antioxidant power (FRAP) in milk compared to those fed no pelleted citrus pulp. Cows fed 18% PCP showed higher monounsaturated FA and lower saturated FA in milk fat compared with cows fed the other diets. The lowest n-6 FA proportion was in milk fat from cows fed control. The present study suggests that pelleted citrus pulp added to 9% to 18% DM increases total polyphenols and flavonoids concentration, and the FRAP in milk.


Flavonoids;Polyphenols;Ferric Reducing Antioxidant Power


  1. Weiss, W. P. and D. J. Wyatt. 2003. Effect of dietary fat and vitamin e on $\alpha$-tocopherol in milk from dairy cows. J. Dairy Sci. 86:3582-3591.
  2. Williams, R. J., J. P. Spencer, and C. Rice-Evans. 2004. Flavonoids: Antioxidants or signalling molecules? Free Radic. Biol. Med. 36:838-849.
  3. Yang, S. L., D. P. Bu, J. Q. Wang, Z. Y. Hu, D. Li, H. Y. Wei, L. Y. Zhou, and J. J. Loor. 2009. Soybean oil and linseed oil supplementation affect profiles of ruminal microorganisms in dairy cows. Animal 3:1562-1569.
  4. Zhu, Q. Y., R. M. Hackman, J. L. Ensunsa, R. R. Holt, and C. L. Keen. 2002. Antioxidative activities of oolong tea. J. Agric. Food Chem. 50:6929-6934.
  5. Santos, F. A. P., M. P. Menezes Junior, J. M. C. Simas, A. V. Pires, and C. M. B. Nussio. 2001. Corn grain processing and its partial replacement by pelleted citrus pulp on performance, nutrient digestibility and blood parameters of dairy cows. Acta. Sci. Anim. Sci. 23:923-931.
  6. Shingfield, K. J., L. Bernard, C. Leroux, and Y. Chilliard. 2010. Role of trans fatty acids in the nutritional regulation of mammary lipogenesis in ruminants. Animal 4:1140-1166.
  7. Shiota, M., H. Konishi, and K. Tatsumi. 1999. Oxidative stability of fish oil blended with butter. J. Dairy Sci. 82:1877-1881.
  8. Singleton, V. L. and J. A. Rossi. 1965. Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. Am. J. Enol. Vitic. 16:144-158.
  9. Solomon, R., L. E. Chase, D. Ben-Ghedalia, and D. E. Bauman. 2000. The effect of nonstructural carbohydrate and addition of full fat extruded soybeans on the concentration of conjugated linoleic acid in the milk fat of dairy cows. J. Dairy Sci. 83:1322-1329.
  10. Sunvold, G. D., H. S. Hussein, G. C. Fahey, N. R. Merchen, and G. A. Reinhart. 1995. In vitro fermentation of cellulose, beet pulp, citrus pulp, and citrus pectin using fecal inoculum from cats, dogs, horses, humans, and pigs and ruminal fluid from cattle. J. Anim. Sci. 73:3639-3648.
  11. Tyrrell, H. F. and J. T. Reid. 1965. Prediction of the energy value of cow's milk. J. Dairy Sci. 48:1215-1223.
  12. Van Soest, P. J. 1994. Nutrional Ecology of the Ruminant. Cornell University Press, Ithaca, NY, USA.
  13. Voltolini, T. V., G. T. Santos, M. A. Zambom, N. P. Ribas, E. E. Muller, J. C. Damasceno, L. C. V. Itavo, and D. R. Veiga. 2001. Influence of lactation stages on the counting of somatic cells of Holstein milk cows and identification of sources of mastitis pathogens in cattle. Acta. Sci. Anim. Sci. 23:961-966.
  14. Mertens, D. R. 2002. Gravimetric determination of amylasetreated neutral detergent fiber in feeds with refluxing in beakers or crucibles: Collaborative study. J. AOAC Int. 85:1217-1240.
  15. Miron, J., E. Yosef, D. Ben-Ghedalia, L. E. Chase, D. E. Bauman, and R. Solomon. 2002. Digestibility by dairy cows of monosaccharide constituents in total mixed rations containing citrus pulp. J. Dairy Sci. 85:89-94.
  16. Murphy, J. J., J. F. Connolly, and G. P. McNeill. 1995. Effects on milk fat composition and cow performance of feeding concentrates containing full fat rapeseed and maize distillers grains on grass-silage based diets. Livest. Prod. Sci. 44:1-11.
  17. National Research Council. 2001. Nutrients Requirements of Dairy Cattle. 7th Ed. National Academies Press, Washington, DC, USA.
  18. Palmquist, D. L., A. D. Beaulieu, and D. M. Barbano. 1993. Feed and animal factors influencing milk fat composition. J. Dairy Sci. 76:1753-1771.
  19. Palmquist, D. L. and T. C. Jenkins. 1980. Fat in lactation rations: Review. J. Dairy Sci. 63:1-14.
  20. Piperova, L. S., B. B. Teter, I. Bruckental, J. Sampugna, S. E. Mills, M. P. Yurawecz, J. Fritsche, K. Ku, and R. A. Erdman. 2000. Mammary lipogenic enzyme activity, trans fatty acids and conjugated linoleic acids are altered in lactating dairy cows fed a milk fat-depressing diet. J. Nutr. 130:2568-2574.
  21. Piquer, O., L. Rodenas, C. Casado, E. Blas, and J. J. Pascual. 2009. Whole citrus fruits as an alternative to wheat grain or citrus pulp in sheep diet: Effect on the evolution of ruminal parameters. Small Rumin. Res. 83:14-21.
  22. Rindsig, R. B. and L. H. Schultz. 1974. Effects of abomasal infusions of safflower oil or elaidic acid on blood lipids and milk fat in dairy cows. J. Dairy Sci. 57:1459-1466.
  23. Rocha Filho, R. R., P. F. Machado, R. D. D'Arce, and J. C. Francisco Jr. 1999. Citrus and corn pulp related to rumen volatile fatty acids production. Sci. Agric. 56:471-477.
  24. Fox, P. F. and P. L. H. McSweeney. 1998. Dairy Chemistry and Biochemistry. Blackie Academic and Professional, Great Britain.
  25. Friedewald, W. T., R. I. Levy, and D. S. Fredrickson. 1972. Estimation of the concentration of low-density lipoprotein cholesterol in plasma, without use of the preparative ultracentrifuge. Clin. Chem. 18:499-502.
  26. Gilaverte, S., I. Susin, A. V. Pires, E. M. Ferreira, C. Q. Mendes, R. S. Gentil, M. V. Biehl, and G. H. Rodrigues. 2011. Diet digestibility, ruminal parameters and performance of Santa Ines sheep fed dried citrus pulp and wet brewer grain. R. Bras. Zootec. 40:639-647.
  27. Grummer, R. R. and D. J. Carroll. 1991. Effects of dietary fat on metabolic disorders and reproductive performance of dairy cattle. J. Anim. Sci. 69:3838-3852.
  28. Hall, M. B., C. C. Larson, and C. J. Wilcox. 2010. Carbohydrate source and protein degradability alter lactation, ruminal, and blood measures. J. Dairy Sci. 93:311-322.
  29. Hwang, S.-L., P.-H. Shih, and G.-C. Yen. 2012. Neuroprotective effects of citrus flavonoids. J. Agric. Food Chem. 60:877-885.
  30. International Organization for Standardization. 1978. Animal and vegetable fats and oils - Preparation of methyl esters of fatty acids, London.
  31. Jenkins, T. C. 1993. Lipid metabolism in the rumen. J. Dairy Sci. 76:3851-3863.
  32. Jenkins, T. C. and M. A. McGuire. 2006. Major advances in nutrition: Impact on milk composition. J. Dairy Sci. 89:1302-1310.
  33. Kalscheur, K. F., B. B. Teter, L. S. Piperova, and R. A. Erdman. 1997. Effect of fat source on duodenal flow of trans-C18:1 fatty acids and milk fat production in dairy cows. J. Dairy Sci. 80:2115-2126.
  34. Kim, S. C., A. T. Adesogan, and J. D. Arthington. 2007. Optimizing nitrogen utilization in growing steers fed forage diets supplemented with dried citrus pulp. J. Anim. Sci. 85:2548-2555.
  35. Borsting, C. F., M. R. Weisbjerg, and T. Hvelplund. 1992. Fattyacid digestibility in lactating cows fed increasing amounts of protected vegetable oil, fish oil or saturated fat. Acta Agric. Scand. Sect. A Anim. Sci. 42:148-156.
  36. Broderick, G. A. and M. K. Clayton. 1997. A statistical evaluation of animal and nutritional factors influencing concentrations of milk urea nitrogen. J. Dairy Sci. 80:2964-2971.
  37. Bu, D. P., J. Q. Wang, T. R. Dhiman, and S. J. Liu. 2007. Effectiveness of oils rich in linoleic and linolenic acids to enhance conjugated linoleic acid in milk from dairy cows. J. Dairy Sci. 90:998-1007.
  38. Buriol, L., D. Finger, E. M. Schmidt, J. M. T. dos Santos, M. R. da Rosa, S. P. Quinaia, Y. R. Torres, H. S. Dalla Santa, C. Pessoa, M. O. de Moraes, L. V. Costa-Lotufo, P. M. P. Ferreira, A. C. H. F. Sawaya, and M. N. Eberlin. 2009. Chemical composition and biological activity of oil propolis extract: An alternative to ethanolic extract. Quim. Nova 32:296-302.
  39. Chilliard, Y., A. Ferlay, R. M. Mansbridge, and M. Doreau. 2000. Ruminant milk fat plasticity: Nutritional control of saturated, polyunsaturated, trans and conjugated fatty acids. Ann. Zootech. 49:181-205.
  40. Cortes, C., D. C. da Silva-Kazama, R. Kazama, N. Gagnon, C. Benchaar, G. T. Santos, L. M. Zeoula, and H. V. Petit. 2010. Milk composition, milk fatty acid profile, digestion, and ruminal fermentation in dairy cows fed whole flaxseed and calcium salts of flaxseed oil. J. Dairy Sci. 93:3146-3157.
  41. Dai, J. and R. J. Mumper. 2010. Plant phenolics: extraction, analysis and their antioxidant and anticancer properties. Molecules 15:7313-7352.
  42. Doner, L. W., G. Becard, and P. L. Irwin. 1993. Binding of flavonoids by polyvinylpolypyrrolidone. J. Agric. Food Chem. 41:753-757.
  43. Fegeros, K., G. Zervas, S. Stamouli, and E. Apostolaki. 1995. Nutritive-value of dried citrus pulp and its effect on milk-yield and milk-composition of lactating ewes. J. Dairy Sci. 78:1116-1121.
  44. AOAC. 1998. Official Methods of Analysis. 16th edn. Association of Official Analytical Chemists, Gaithersburg, MD, USA.
  45. Arthington, J. D., W. E. Kunkle, and A. M. Martin. 2002. Citrus pulp for cattle. Vet. Clin. North Am. Food Anim. Pract. 18:317-326.
  46. Assis, A. J., J. M. D. Campos, S. D. Valadares, A. C. de Queiroz, R. D. Lana, R. F. Euclydes, J. M. Neto, A. L. R. Magalhaes, and S. D. Mendonca. 2004. Citrus pulp in diets for milking cows. Intake of nutrients, milk production and composition. R. Bras. Zootec. 33:242-250.
  47. Bampidis, V. A. and P. H. Robinson. 2006. Citrus by-products as ruminant feeds: A review. Anim. Feed Sci. Technol. 128:175-217.
  48. Bateman, H. G. and T. C. Jenkins. 1998. Influence of soybean oil in high fiber diets fed to nonlactating cows on ruminal unsaturated fatty acids and nutrient digestibility. J. Dairy Sci. 81:2451-2458.
  49. Bauman, D. E. and J. M. Griinari. 2001. Regulation and nutritional manipulation of milk fat: low-fat milk syndrome. Livest. Prod. Sci. 70:15-29.
  50. Bauman, D. E. and J. M. Griinari. 2003. Nutritional regulation of milk fat synthesis. Annu. Rev. Nutr. 23:203-227.
  51. Baumgard, L. H., B. A. Corl, D. A. Dwyer, A. Saebo, and D. E. Bauman. 2000. Identification of the conjugated linoleic acid isomer that inhibits milk fat synthesis. Am. J. Physiol. Regul. Integr. Comp. Physiol. 278:179-184.

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