Effects of Mustard Seeds and Powder on In vitro Ruminal Fermentation Characteristics and Methane Production

겨자종자와 겨자분의 첨가가 반추위 발효성상과 메탄생성에 미치는 영향

  • 이강연 (농촌진흥청 국립축산과학원) ;
  • 김경훈 (농촌진흥청 국립축산과학원) ;
  • 백열창 (농촌진흥청 국립축산과학원) ;
  • 옥지운 (농촌진흥청 국립축산과학원) ;
  • 설용주 (농촌진흥청 국립축산과학원) ;
  • 한기준 (농촌진흥청 국립축산과학원) ;
  • 박근규 (건국대학교 동물자원센터) ;
  • 류호태 (자연과기술(주)) ;
  • 이상석 (순천대학교 동물자원과학과) ;
  • 전체옥 (중앙대학교 자연과학대학) ;
  • 오영균 (농촌진흥청 국립축산과학원)
  • Received : 2012.04.30
  • Accepted : 2013.02.26
  • Published : 2013.02.28


The purpose of this study was to investigate the effects of mustard, which contains allyl isothiocyanate, on ruminal fermentation and methane emission in vitro. To this end, diluted ruminal fluid(30ml) was incubated anaerobically at $39^{\circ}C$ or 6, 12, and 24 h with or without seeds or powdered mustard. Either mustard seed or powdered mustard was weighed and serially (0, 3.33, 5.00, 6.67, and 8.33 g/L) mixed with ruminal fluid. Ammonia-N was increased (P < 0.05) by mustard treatment in a dose dependent manner. Regardless of concentration or form, mustard increased (P < 0.05) total VFA content but decreased (P < 0.01) pH compared to control group. Molar proportion of acetate (A) was decreased (P < 0.05) whereas propionate (P) was increased (P < 0.05) by mustard treatment, thereby A:P ratio was decreased (P < 0.05) compared to control group. Total gas production was increased (P < 0.01) in a linear manner by mustard treatment compared to control group. There was no effect of mustard powder, except 8.33 g/L level at 6 h, on methane emission. However, at 24 h, methane emission was reduced (P < 0.05) by 4.77% and 11.54% with 6.67 g/L and 8.33 g/L of mustard seeds supplementation, respectively. Altogether, these results suggest that mustard seeds containing allyl isothiocyanate may reduce methane production without disturbing ruminal fermentation.


Mustard;Methane;Ruminal fermentation


  1. Agarwal, N., Shekhar, C., Kumar, R., Chaudhary, L. C. and Kamra, D. N. 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.
  2. Amlan K. Patra. And Jyotisna Saxena. 2010. A new perspective on the use of plant secondary metabolites in hibit methanogenesis in the rumen. Phytocheminstry. 71:1198-1222.
  3. Amrish Kumar Tyagi. 2002. Influence of water soaking of mustard cake on glucosinolate hydrolysis. Anim. Feed Sci. Technol. 99:215-219.
  4. Blaxter, K. L. and Clapperton, J. L. 1965. Prediction of the amount of methane produced by ruminants. British Journal of Nutrition 19:511-522.
  5. Bodas, R., Lopez, S., Fernandez. M., Garcia-Gonzalez, R., Rodriguez, A. B., Walace, R. J. and Gonzalez, J. S. 2008. In vitro screening of the potential of numerous plant spicies as antimethanogenic feed additives for ruminants. Anim. Feed Sci. Technol. 145:245-258.
  6. Cardozo, P. W., Calsamiglia, S., Ferret, A. and Kamel, C. 2005. Screening for the effects of natural plant extracts at different pH on in vitro rumen microbial fermentation of a high-concentrate diet for beef cattle. J. Anim. Sci. 83:2572-2579.
  7. Chaney, A. L. and Marbacch, E. P. 1962. Modification reagents for determination of urea and ammoina. Clin. Chem. 8:130.
  8. Chaves, A. V., He, M. L., Yang, W. Z., Hristov, A. N., McAllister, T. A. and Benchaar, C. 2008. Effects of essential oils on proteolytic, deaminative and methanogenic acitivities of mixed ruminal bacteria. Can. J. Anim. Sci. 89:97-104.
  9. Demeyer, D. and Van Nevel, C. J. 1975. Methanogenesis, and intergrated part of carbohydrate fermentation and its control. Pages 366-382 in Digestion and Metabolism in the Ruminant. L. W. McDonald and A. C. I. Warner, ed. Univ. of New England Publishing Unit, Armidale, Australia.
  10. Ferorak, P. M. and Hrwdey, S. E. 1983. A simple apparatus for measuring gas production by methanogenic culture in serum bottl. Environ. Technol. Lett. 4:268.
  11. Garcia-Gonzalez, R., Lopez, S., Fernadez, M., Bodas, R. and Gonzalez, J. S. 2008. Screening the acticity of plants and spices for decreasing ruminal methane production in vitro. Anim. Feed Sci. Technol. 147:36-52.
  12. Goodrich, R. J., Garnett, J. E., Gast, D. R., Kirick, M. A., Larson, D. A. and Meiske, J. C. 1984. Influence of monensin on the performance of cattle. J. Anim. Sci. 58:1484-1498.
  13. Hino, T. and Russell, J. B. 1985. Effect of reducing-equivalent disposal and NADH/NAD on deamination of amino acids by intact rumen microorganisms and their cell extracts. Appl. Environ. Microbiol. 50(6):1368-1374.
  14. IPCC (Intergovernmnet Panel on Climate Change). 2001. Climate change 2001. The scientfic basis. Cambridge, UK: Cambridge University Press.
  15. Johnson, D. E., Wood, A. S., Stone, J. B. and Moren, E. T. Jr. 1972. Some effects of methane inhibition in ruminants (steers). Ca. J. Ani. Sci. 52: 703-708.
  16. Johnson, K. A. and Johnson, D. E. 1995. Methane emission from cattle. J. Anim. Sci. 73:2483-2492.
  17. Kamel, C., Greathead, H. M. R., Tejido, M. L., Ranilla, M. J. and Carro, M. D. 2008. Effects of allicin and diallyl disulfide on in vitro rumen fermentation of a mixed diet. Anim. Feed Sci. Technol. 145:351-363.
  18. Kongmun, P., Wanapat, M., Pakdee, P. and Navanukraw, C. 2010. Effect of cocount oil and garlic powder o in vitro fermentation using gas production technique. Licest Sci. 127:38-44.
  19. Kumar, R., Kamra, D. N., Agrawal, N. and Chaudhary, L. C. 2009. Effect of eucalyptus (Eucalyptus globulus) oil on in vitro methanogenesis and fermentation of feed with buffalo rumen liquor. Anim. Nutr. Feed Technol. 9:237-243.
  20. Lila, Z. A., Mohammed, N., Kanda, S., Kamada, T. and Itabashi, H. 2003. Effect of ${\alpha}$-cyclodextrin allyl isothiocyanate on ruminal microbial methane production in vitro. Anim. Sci. J. 74:321-326.
  21. Lin, C. M., Kim, J., Du, W. X. and Wei, C. I. 2000a. Bactericidal activity of isothiocyanate against pathogens on fresh produce. J. Food protiction. 63:25-30.
  22. Lin, C. M., Preston, J. F. and Wei, C. I. 2000b. Antibacterial mechanism of allyl isothiocyanate. J. Food Protection. 63:727-734.
  23. McDougall, E. I. 1947. Studies on ruminant saliva. I. The composition and output of sheep's salive. Biochem. J. 43:99-109.
  24. Mithen, R. 2006. Sulphur-contaning compounds. In: Crozier, A., Clifford, M. N., Ashihara, H. (Eds.), Plant secondary metabolites: Occurrence, Structure and Role in the human diet. Blackwell publishing, Chennai, India. pp. 25-46.
  25. Mohammed, N., Ajisaka, N., Lila, Z. A., Koji Hara., Mikuni, K., Hara, K., Kanda, S. and Itabashi, H. 2004. Effect of Japanese horeradish oil on methane production and ruminal fermentation in vitro and in steers. J. Anim. Sci. 82:1839-1846.
  26. Newbold, C. J., Lassalas, B. and Jouany, P. Jouany. 1995. The importance of methanogens associated with ciliate protozoa in ruminal met hane production in vitro. Microbiology. 21:230-234.
  27. Ohene-Adjei, S., Chaves, A. V., McAllister, T. A., Benchaar, C., Teather, R. M. and Forster, R. J. 2008. Evidunce of increased diversity of methanogenic archaea with plant extract supplementation. Microial. Ecol. 56:234-242.
  28. Patra, A. K., Kamra, D. N. and Agarwal, N. 2006a. Effect of plant extracts on in vitro methanogenesis, enzyme activities and fermentation of feed in rumen liquor of buffalo. Anim. Feed. Sci. Technol. 128:276-291.
  29. Patra, A. K., Kamra, D. N. and Agarwal, N. 2006b. Effect of spices on rumen fermenation, methanogenesis and protozoa counts in in vitro gas productio test. Int. Comgress Ser. 1293:176-179.
  30. Rufener, W. H. Jr. and Wolin, M. J. 1968. Effect of CCl4 on methane and volatile acid production in continuous culture of rumen organisms and sheep rumen. Microbiology. 16:1955-1956.
  31. Rune, BjOrkman. Interaction between proteins and glucosinolate isothiocyanates and oxazolidinethinones from Brassica napus seed. 1973. Phytochemistry. 12:1585-1590.
  32. SAS. 2002. SAS User's Guide : Statistics, Version 9.1 Edition. SAS Inst., Inc., Cary, NC.
  33. Son, M. H. and Lee, J. Y. 2006. Standardization of processing conditions of mustard powder and mustard oil for quality improvement. The Korean Journal of culinary research. 12(4):131-139.
  34. The Government of the Republic of Korea. 2003. second National Communication of the Republic of Korea Under the United Nations Framework Convention on Climate Change.
  35. Trei, J. E., Parish, R. C., Singh, Y. K. and Scott, G. C. 1971. Effect of methane inhibitors on rumen metabolism and feedlot performance of sheep. J. Dairy Sci. 54:536-540.
  36. Tyagi, A. K. and Singhal, K. K. 1998. Effect of mustard oil and glucosinolate on rumen fermentation. Indian J. Anim. Nutr. 16:12-18.
  37. Van Nevel, C. J., Henderickx, H. K., Demeyer, D. I. and Martin, J. 1969. Effect of chloral hydrate on methane and propionic acid in the rumen. Microbiology. 17:695-700.
  38. Zebeli, Q., Tafaj, M., Weber, I., Steingass, H. and Drochner, W. 2008. Effects of dietary ferage particle size and concentarate level on fermentation profile, in vitro degradation characteristics and conentration of liquid- or solid- associated bacterial mass in the rumen of dairy cows. J. Anim. Feed Sci. Technol. 140:307-325.