Effects of Eucalyptus Crude Oils Supplementation on Rumen Fermentation, Microorganism and Nutrient Digestibility in Swamp Buffaloes

  • Thao, N.T. (Department of Animal Science and Veterinary Medicine, An Giang University) ;
  • Wanapat, M. (Tropical Feed Resources Research and Development Center, (TROFREC), Department of Animal Science, Faculty of Agriculture, Khon Kaen University) ;
  • Cherdthong, A. (Tropical Feed Resources Research and Development Center, (TROFREC), Department of Animal Science, Faculty of Agriculture, Khon Kaen University) ;
  • Kang, S. (Tropical Feed Resources Research and Development Center, (TROFREC), Department of Animal Science, Faculty of Agriculture, Khon Kaen University)
  • Received : 2013.05.29
  • Accepted : 2013.11.01
  • Published : 2014.01.01


This study was conducted to investigate the effects of eucalyptus (E. Camaldulensis) crude oils (EuO) supplementation on voluntary feed intake and rumen fermentation characteristics in swamp buffaloes. Four rumen fistulated swamp buffaloes, body weight (BW) of $420{\pm}15.0$ kg, were randomly assigned according to a $2{\times}2$ factorial arrangement in a $4{\times}4$ Latin square design. The dietary treatments were untreated rice straw (RS) without EuO (T1) and with EuO (T2) supplementation, and 3% urea-treated rice straw (UTRS) without EuO (T3) and with EuO (T4) supplementation. The EuO was supplemented at 2 mL/h/d in respective treatment. Experimental animals were kept in individual pens and concentrate mixture was offered at 3 g/kg BW while roughage was fed ad libitum. Total dry matter and roughage intake, and apparent digestibilites of organic matter and neutral detergent fiber were improved (p<0.01) by UTRS. There was no effect of EuO supplementation on feed intake and nutrient digestibility. Ruminal pH and temperature were not (p>0.05) affected by either roughage sources or EuO supplementation. However, buffaloes fed UTRS had higher ruminal ammonia nitrogen and blood urea nitrogen as compared with RS. Total volatile fatty acid and butyrate proportion were similar among treatments, whereas acetate was decreased and propionate molar proportion was increased by EuO supplementation. Feeding UTRS resulted in lower acetate and higher propionate concentration compared to RS. Moreover, supplementation of EuO reduced methane production especially in UTRS treatment. Protozoa populations were reduced by EuO supplementation while fungi zoospores remained the same. Total, amylolytic and cellulolytic bacterial populations were increased (p<0.01) by UTRS; However, EuO supplementation did not affect viable bacteria. Nitrogen intake and in feces were found higher in buffaloes fed UTRS. A positive nitrogen balance (absorption and retention) was in buffaloes fed UTRS. Supplementation of EuO did not affect nitrogen utilization. Both allantoin excretion and absorption and microbial nitrogen supply were increased by UTRS whereas efficiency of microbial protein synthesis was similar in all treatments. Findings of present study suggested that EuO could be used as a feed additive to modify the rumen fermentation in reducing methane production both in RS and UTRS. Feeding UTRS could improve feed intake and efficiency of rumen fermentation in swamp buffaloes. However, more research is warranted to determine the effect of EuO supplementation in production animals.


Eucalyptus Oils;Rumen Fermentation;Nutrients Digestibility;Rice Straw;Swamp Buffalo


  1. Wanapat, M. 1985. Improving rice straw qulity as ruminant feed by urea-treatment in Thailand, pp. 147-175. In: Relevance of crop Residues as Animal Feed in Developing Countries (Ed. M. Wanapat and C. Devendra). Funny Press: Bangkok.
  2. Wanapat, M. 1990. Nutritional aspects of ruminant production in Southeast Asia with special reference to Thailand. Funny Press, Ltd., Bangkok, Thailand.
  3. Wanapat, M., S. Polyorach, K. Boonnop, C. Mapato, and A. Cherdthong. 2009. Effects of treating rice straw with urea or urea and calcium hydroxide upon intake, digestibility, rumen fermentation and milk yield of dairy cows. Livest. Sci. 125:238-243.
  4. Wang C. J., S. P. Wang, and H. Zhou. 2009. Influences of flavomycin, ropadiar and saponin on nutrient digestibility, rumen fermentation and methane emission from sheep. Anim. Feed Sci. Technol. 148:157-166.
  5. Yang, W. Z., B. N. Ametaj, C. Benchaar, M. L. He, and K. A. Beauchemin. 2010. Cinnamadehyde in feedlot cattle diets: Intake, growth performance, carcass characteristics and blood metabolites. J. Anim. Sci. 88:1082-1092.
  6. Yang, W. Z., C. Benchaar, B. N. Ametaj, A. V. Chaves, M. L. He, and T. A. McAllister. 2007. Effects of garlic and juniper berry essential oils on ruminal fermentation and on the site and extent of digestion in lactating cows. J. Dairy Sci. 90:5671-5681.
  7. AOAC. 1995. Official methods of analyses, 16th ed. Animal Feeds: Association of Official Analytical Chemists, VA, USA.
  8. Beauchemin, K. A. and S. N. McGinn. 2006. Methane emissions from beef cattle: effects of fumaric acid, essential oil, and canola oil. J. Anim. Sci. 84:1489-1496.
  9. Benchaar, C., H. V. Petit, R. Berthiaume, T. D. Whyte, and P. Y. Chouinard. 2006a. Effects of addition of essential oils and monensin premix on digestion, ruminal fermentation, milk production and milk composition in dairy cows. J. Dairy Sci. 89:4352-4364.
  10. Benchaar, C., J. L. Duynisveld, and E. Charmley. 2006b. Effects of monensin and increasing dose levels of a mixture of essential oil compounds on intake, digestion and growth performance of beef cattle. Can. J. Anim. Sci. 86:91-96.
  11. Bencharr, C., H. V. Petit, R. Berthiaume, D. R. Ouellet, J. Chiquette, and P. Y. Chouinard. 2007. Effects of essential oils on digestion, ruminal fermentation, rumen microbial populations, milk production, and milk composition in dairy cows fed alfalfa silage or corn silage. J. Dairy. Sci. 90:886-897.
  12. Bunting, L. D., J. A. Boling, C. T. MacKown, and R. B. Muntifering. 1987. Effect of dietary protein level on nitrogen metabolism in lambs: Studies using 15N-nitrogen. J. Anim. Sci. 64:855-867.
  13. Busquet, M., S. Calsamiglia, A. Ferret, and C. Kamel. 2003. Efecto del extracto de ajo y/o cinnamaldehi do sobre la produccion, composicion y residuos en vacas de alta produccion. ITEA Vol-Extra. 24:756-758.
  14. Agarwal, N., C. Shekhar, R. Kumar, L. C. Chaudhary, and D. N. Kamra. 2009. Effect of peppermint (Mentha piperita) oil on in vitro methanogenesis and fermentation of feed with buffalo rumen liquor. Anim. Feed Sci. Technol. 148:321-327.
  15. Ando, S., T. Nishida, M. Ishida, K. Hosoda, and E. Bayaru. 2003. Effect of peppermint feeding on the digestibility, ruminant fermentation and protozoa. Livest. Prod. Sci. 82:245-248.
  16. Busquet, M., S. Calsamiglia, A. Ferret, and C. Kamel. 2006. Plant extracts affect in vitro rumen microbial fermentation. J. Dairy Sci. 89:761-771.
  17. Busquet, M., S. Calsamiglia, A. Ferret, M. D. Carro, and C. Kamel. 2005. Effects of garlic oil and four of its compounds on rumen microbial fermentation. J. Dairy Sci. 88:4393-4404.
  18. Cardozo, P. W., S. Calsamiglia, A. Ferret, and C. Kamel. 2006. Effects of alfalfa extract, anise, capsicum and a mixture of cinnamaldehyde and eugenol on ruminal fermentation and protein degradation in beef heifers fed a high concentrate diet. J. Anim. Sci. 84:2801- 2808.
  19. Chantalakhana, C. 2001. Contribution of Water Buffaloes in Rural Development. Proceeding Regional Workshop on Water Buffalo Development, Thongtarin Hotel, Surin, Thailand, 8-10 Februarry, 2001.
  20. Chaves, A. V., K. Stanford, M. E. R. Dugan, L. L. Gibson, T. A. McAllister, F. Van Herk, and C. Benchaar. 2008a. Effects of cinnamaldehyde, garlic and juniper berry essential oils on rumen fermentation, blood metabolites, growth performance, and carcass characteristics of growing lambs. Livest. Sci. 117:215-224.
  21. Chaves, A. V., M. L. He, W. Z. Yang, A. N. Hristov, T. A. McAllister, and C. Benchaar. 2008b. Effects of essential oils on proteolytic, deaminative and methanogenic activities of mixed ruminal bacteria. Can. J. Anim. Sci. 88:117-122.
  22. Chen, X. B. and M. J. Gomes. 1995. Estimation of microbial protein supply to sheep and cattle based on urinary excretion of purine derivative An overview of the technique details. Occasional publication 1992. International Feed Resources Unit, Rowett Research Institute, Aberdeen, UK.
  23. Chen, X. B., D. J. Kyle, and E. R. Orskov. 1993. Measurement of allantoin in urine and plasma by high performance liquid chromatography with pre-column derivatization. J. Chromatogr. B. Biomed. Sci. Appl. 617:241-247.
  24. Chen, X. L., J. K. Wang, Y. M. Wu, and J. X. Liu. 2008. Effects of chemical treatments of rice straw on rumen fermentation characteristics, fibrolytic enzyme activities and populations of liquid- and solid-associated ruminal microbes in vitro. Anim. Feed Sci. Technol. 141:1-14.
  25. Crichton, N. 1999. Information point: Tukey's multiple Comparison test. Blackwell Science Ltd, J. Clinical Nursing, 8:299-304.
  26. Crocker, C. L. 1967. Rapid determination of urea nitrogen in serum or plasma without deproteinization. Am. J. Med. Technol. 33:361-365.
  27. Devendra, C. 1992. Nutrition of swamp buffalo. In: Buffalo Production, World Animal Science, C6. (Ed. N. M. Tulloh and J. H. G. Holmes). Elsevier. Tokyo.
  28. Fadel-Elseed, A. M. A., J. Sekine, M. Hishinuma, and K. Hamana. 2003. Effects of ammonia, urea plus calcium hydroxide and animal urine treatments on chemical composition and in sacco degradability of rice straw. Asian-Aust. J. Anim. Sci. 16:368-373.
  29. Galyean, M. 1989. Laboratory procedure in animal nutrition research. Department of animal and range science. New Mexico State University, USA.
  30. Giannenas, I., J. Skoufos, C. Giannakopoulos, M. Wiemann, O. Gortzi, S. Lalas, and I. Kyriazakis. 2011. Effect of essential oils on milk production, milk composition, and rumen microbiota in Chios dairy ewes. J. Dairy. Sci. 94:5569-5577.
  31. Goto, M., Y. Yokoe, K. Takabe, S. Nishikawa, and O. Morita. 1993. Effects of gaseous ammonia on chemical and structural features of cell walls in spring barley straw. Anim. Feed Sci. Technol. 40:207-221.
  32. Helander, I. M., H. Alakomi, K. Latva-Kala, T. Mattila-Sandholm, I. Pol, E. J. Smid, L. G. M. Gorris, and A. Wright. 1998. Characteritzation of the action of selected essential oil components on gram-negative bacteria. J. Agric. Food Chem. 46:3590-3595.
  33. Hess, H. D., C. E. Lascano, and H. Florez. 2000. Blood and milk urea nitrogen as a tool to monitor the protein nutrition of cattle under tropical conditions. Institute of Animal Sciences, Animal Nutrition, ETH, Zurich, Switzerland.
  34. Hoover, W. H. and S. R. Stokes. 1991. Balancing carbohydrates and proteins for optimum rumen microbial yield. J. Dairy Sci. 74:3630-3644.
  35. Hungate, R. E. 1969. A role tube method for cultivation of strict anaerobes. Method in mMicrobiology (Ed. J. R. Norris and D. W. Ribbons). New York, Academic. NY. 313.
  36. Kebreab, E., G. D. Cruz, P. H. Hai, N. Anatassok, S. Polyorach, P. Beelen, and H. D. Rosa. 2012. Methane emission profile in dairy heifers feed rice straw. In Proc. The 1st International Conference on Animal Nutrition and Environment. Khon Kaen, Thailand.
  37. Khejornsart, P., M. Wanapat, and P. Rowlinson. 2011. Diversity of anaerobic fungi and rumen fermentation characteristic in swamp buffalo and beef cattle fed on different diet. Livest. Sci. 139:230-236.
  38. Kongmun, P., M. Wanapat, P. Pakdee, C. Navanukraw, and Z. Yu. 2011. Manipulation of rumen fermentation and ecology of swamp buffalo by coconut oil and garlic powder supplementation. Livest. Sci. 135:84-92.
  39. Kongmun, P., M. Wanapat, P. Pakdee, and C. Navanukraw. 2010. Effect of coconut oil and garlic powder on in vitro fermentation using gas production technique. Livest. Sci. 127:38-44.
  40. Leng, R. A. 1993. The impact of livestock development on environmental change. FAO Corporate Documentary Repository, pp:1-14.
  41. Liang, J. B., M. Matsumoto, and B. A. Young. 1994. Purine derivative excretion and rumen microbial yield in Malaysian cattle and swamp buffalo. Anim. Feed Sci. Technol. 47:189-199.
  42. Malecky, M., L. P. Broudiscou, and P. Schmidely. 2009. Effects of two levels of monoterpene blend on rumen fermentation, terpene and nutrient flows in the duodenum and milk production in dairy goats. Anim. Feed Sci. Technol. 154:24-35.
  43. McInstosh, F. M., P. Williams, R. Losa, R. J. Wallace, D. A. Beever, and C. J. Newbold. 2003. Effect of essential oils on ruminal microorganisms and their protein metabolism. Appl. Environ. Microbiol. 69:5011-5014.
  44. Mohammed, N., N. Ajisaka, Z. A. Lila, K. Mikuni, S. Kanda, and H. Itabashi. 2004. Effects of Japanese horseradish oil on methane production and ruminal fermentation in vitro and in steers. J. Anim. Sci. 82:1839-1846.
  45. Moss, A. R., J. P. Jouan, and J. Ne wbold. 2000. Methane production by ruminants: its contribution to global warming. Ann. Zootech. 49:231-253.
  46. Newbold, C. J., F. M. McInstosh, P. Williams, R. Losa, and J. Wallace. 2004. Effects of a specific blend of essential oils compounds on rumen fermentation. Anim. Feed Sci. Technol. 114:105-112.
  47. Patra, A. K. 2011. Effects of essential oils on rumen fermentation, microbial ecology and ruminant production. Asian J. Anim. Vet. Adv. 6:416-428.
  48. Patra, A. K., D. N. Kamra, and N. Agarwal. 2010. Effects of extracts of spices on rumen methanogenesis, enzyme activity and fermentation of feeds in vitro. J. Sci. Food Agric. 90:511-520.
  49. Perdok, H. and R. A. Leng. 1989. Rumen ammonia requirements for efficient digestion and intake of straw by cattle. In: The Roles of Protozoa and Fungi in ruminant Digestion (Ed. J. V. Nolan and R. A. Leng, Demeyer). Penambul Books, Armidale, Australia.
  50. Preston, T. P. and R. A. Leng. 1987. Matching ruminant production systems with available resources in the tropics and sub-tropics. Penambul Books, Armidale, N.S.W. Australia.
  51. Russell, J. B., H. J. Strobel, and G. Chen. 1988. Enrichment and isolation of a ruminal bacterium with a very high specific activity of ammonia production. Appl. Environ. Microbiol. 54:872-877.
  52. Sallam, S. M. A., I. C. S Bueno, M. E. A. Nasser, and A. L. Abdalla. 2010. Effect of eucalyptus (Eucalyptus citriodora) fresh or residue leaves on methane emission in vitro. Ital. J. Anim. Sci. 9:e58.
  53. Sallam, S. M. A., M. E. A. Nasser, R. C. Araujo, and A. L. Abdalla. 2009. Methane emission in vivo by sheep consuming diet with different levels of eucalyptus essential oil. In Proc. FAO/IAEA Int. Symp. On sustainable improvement of animal production and health, Vienna, Austria.
  54. Samuel, M., S. Sagathewan, J. Thomas, and G. Mathen. 1997. An HPLC method for estimation of volatile fatty acids of ruminal fluid. Indian J. Anim. Sci. 67:805-807.
  55. Santos, M. B., P. H. Robinson, P. Williams, and R. Losa. 2010. Effect of addition of an essential oil complex to the diet of lactating dairy cows on whole tract digestion of nutrients and productive performance. Anim. Feed Sci. Technol. 157:64-71.
  56. SAS. 1998. SAS/STAT User's Guid 6. 12th ed. SAS Institue Inc., Cary, Nort Carolina.
  57. Shibata, M., F. Terada, M. Kurihara, T. Nishida, and K. Iwasaki. 1993. Estimation of methane production in ruminants. Anim. Sci. Technol. 64:790-796.
  58. Tatsouka, N., K. Hara, K. Mikuni, K. Hara, H. Hashimoto, and H. Itabashi. 2008. Effects of the essential oil cyclodextrin complexes on ruminal methane production in vitro. Anim. Sci. J. 79:68-75.
  59. Tekippe, J. A., A. N. Hristov, K. S. Heyler, T. W. Cassidy, V. D. Zheljazkov, J. F. S. Ferreira, S. K. Karnati, and G. V. Varga. 2011. Rumen fermentation and production effects of Origanum vulgare L. leaves in lactating dairy cows. J. Dairy. Sci. 94:5065-5079.
  60. Van Soest, P. J., J. B. Robertson, and B. A. Lewis. 1991. Methods of dietary fiber, neutral detergent fiber and non-starch carbohydrates in relation to animal nutrition. J. Dairy Sci. 74:3583-3597.
  61. Wallace, R. J. 2004. Antimicrobial properties of plant secondary metabolites. Proc. Nutr. Soc. 63:621-629.
  62. Wallace, R. J., N. R. McEwan, F. M. McIntosh, B. Teferedegne, and C. J. Newbold. 2002. Natural products as manipulators of rumen fermentation. Asian-Aust. J. Anim. Sci. 15:1458-1468.
  63. Wanapa M., P. Khejornsart, P. Pakdee, and S. Wanapat. 2008. Effect of supplementation of garlic powder on rumen ecology and digestibility of nutrients in ruminants. J. Sci. Food Agric. 88:2231-2237.
  64. Wanapat M. and O. Pimpa. 1999. Effect of ruminal ammonia nitrogen levels on ruminal fermentation, purine derivatives, digestibility and rice straw intake in swamp buffaloes. Asian-Aus. J. Anim. Sci. 12:904-907.

Cited by

  1. Effects of Supplementation of Eucalyptus (E. Camaldulensis) Leaf Meal on Feed Intake and Rumen Fermentation Efficiency in Swamp Buffaloes vol.28, pp.7, 2015,
  2. Enrichment of Animal Diets with Essential Oils—A Great Perspective on Improving Animal Performance and Quality Characteristics of the Derived Products vol.4, pp.2, 2017,