Effect of Soybean Meal and Soluble Starch on Biogenic Amine Production and Microbial Diversity Using In vitro Rumen Fermentation

  • Jeong, Chang-Dae (Ruminant Nutrition and Anaerobe Laboratory, Department of Animal Science and Technology, Sunchon National University) ;
  • Mamuad, Lovelia L. (Ruminant Nutrition and Anaerobe Laboratory, Department of Animal Science and Technology, Sunchon National University) ;
  • Kim, Seon-Ho (Ruminant Nutrition and Anaerobe Laboratory, Department of Animal Science and Technology, Sunchon National University) ;
  • Choi, Yeon Jae (Ruminant Nutrition and Anaerobe Laboratory, Department of Animal Science and Technology, Sunchon National University) ;
  • Soriano, Alvin P. (Ruminant Nutrition and Anaerobe Laboratory, Department of Animal Science and Technology, Sunchon National University) ;
  • Cho, Kwang Keun (Department of Animal Resource Technology, Gyeongnam National University of Science and Technology) ;
  • Jeon, Che-Ok (Department of Life Science, Chung-Ang University) ;
  • Lee, Sung Sil (Division of Applied Science, Graduate School of Gyeongsang National University, IALS) ;
  • Lee, Sang-Suk (Ruminant Nutrition and Anaerobe Laboratory, Department of Animal Science and Technology, Sunchon National University)
  • Received : 2014.07.23
  • Accepted : 2014.09.29
  • Published : 2015.01.01


This study was conducted to investigate the effect of soybean meal (SM) and soluble starch (SS) on biogenic amine production and microbial diversity using in vitro ruminal fermentation. Treatments comprised of incubation of 2 g of mixture (expressed as 10 parts) containing different ratios of SM to SS as: 0:0, 10:0, 7:3, 5:5, 3:7, or 0:10. In vitro ruminal fermentation parameters were determined at 0, 12, 24, and 48 h of incubation while the biogenic amine and microbial diversity were determined at 48 h of incubation. Treatment with highest proportion of SM had higher (p<0.05) gas production than those with higher proportions of SS. Samples with higher proportion of SS resulted in lower pH than those with higher proportion of SM after 48 h of incubation. The largest change in $NH_3$-N concentration from 0 to 48 h was observed on all SM while the smallest was observed on exclusive SS. Similarly, exclusive SS had the lowest $NH_3$-N concentration among all groups after 24 h of incubation. Increasing methane ($CH_4$) concentrations were observed with time, and $CH_4$ concentrations were higher (p<0.05) with greater proportions of SM than SS. Balanced proportion of SM and SS had the highest (p<0.05) total volatile fatty acid (TVFA) while propionate was found highest in higher proportion of SS. Moreover, biogenic amine (BA) was higher (p<0.05) in samples containing greater proportions of SM. Histamines, amine index and total amines were highest in exclusive SM followed in sequence mixtures with increasing proportion of SS (and lowered proportion of SM) at 48 h of incubation. Nine dominant bands were identified by denaturing gradient gel electrophoresis (DGGE) and their identity ranged from 87% to 100% which were mostly isolated from rumen and feces. Bands R2 (uncultured bacterium clone RB-5E1) and R4 (uncultured rumen bacterium clone L7A_C10) bands were found in samples with higher proportions of SM while R3 (uncultured Firmicutes bacterium clone NI_52), R7 (Selenomonas sp. MCB2), R8 (Selenomonas ruminantium gene) and R9 (Selenomonas ruminantium strain LongY6) were found in samples with higher proportions of SS. Different feed ratios affect rumen fermentation in terms of pH, $NH_3$-N, $CH_4$, BA, volatile fatty acid and other metabolite concentrations and microbial diversity. Balanced protein and carbohydrate ratios are needed for rumen fermentation.


16S rDNA Denaturing Gradient Gel Electrophoresis;Biogenic Amine;In vitro Rumen Fermentation;Soybean Meal and Soluble Starch Ratio


Grant : Cooperative Research Program for Agriculture Science and Technology Development

Supported by : Rural Development Administration


  1. Aschenbach, J. R. and G. Gabel. 2000. Effect and absorption of histamine in sheep rumen: Significance of acidotic epithelial damage. J. Anim. Sci. 78:464-470.
  2. Chaney, A. L. and E. P. Marbach. 1962. Modified reagents for determination of urea and ammonia. Clin. Chem. 8:130-132.
  3. Fenderson, C. L. and W. G. Bergen. 1976. Effect of excess dietary protein on feed intake and nitrogen metabolism in steers. J. Anim. Sci. 42:1323-1330.
  4. Dawson, L. E. R. and C. S. Mayne. 1996. The effect of intraruminal infusions of amines and gamma amino butyric acid on rumen fermentation parameters and food intake of steers offered grass silage. Anim. Feed Sci. Technol. 63:35-49.
  5. Dawson, L. E. R. and C. S. Mayne. 1997. The effect of infusion of putrescine and gamma amino butyric acid on the intake of steers offered grass silage containing three levels of lactic acid. Anim. Feed Sci. Technol. 66:15-29.
  6. Demeyer, D. I. and C. J. Van Nevel. 1975. Methanogenesis and integrated part of carbohydrate fermentation and its control. Digestion and Metabolism in the Ruminant The University of New England Publishing Unit, Armidale, NSW, Australia.
  7. Fusi, E., L. Rossi, R. Rebucci, F. Cheli, A. Di Giancamillo, C. Domeneghini, L. Pinotti, V. Dell'Orto, and A. Baldi. 2004. Administration of biogenic amines to Saanen kids: Effects on growth performance, meat quality and gut histology. Small Rumin. Res. 53:1-7.
  8. Gagen, E. J., P. Mosoni, S. Denman, R. Jassim, C. McSweeney, and E. Forano. 2012. Methanogen colonisation does not significantly alter acetogen diversity in lambs isolated 17 h after birth and raised aseptically. Microb. Ecol. 64:628-640.
  9. Getachew, G., E. J. DePeters, and P. H. Robinson. 2004. In vitro gas production provides effective method for assessing ruminant feeds. Calif. Agric. 58:54.
  10. Han, S.-K., S.-H. Kim, and H.-S. Shin. 2005. UASB treatment of wastewater with VFA and alcohol generated during hydrogen fermentation of food waste. Process Biochem. 40:2897-2905.
  11. Hane, B. G., K. Jager, and H. G. Drexler. 1993. The pearson product-moment correlation coefficient is better suited for identification of DNA fingerprint profiles than band matching algorithms. Electrophoresis 14:967-972.
  12. Mamuad, L., S. H. Kim, C. D. Jeong, Y. J. Choi, C. O. Jeon, and S.-S. Lee. 2014. Effect of fumarate reducing bacteria on in vitro rumen fermentation, methane mitigation and microbial diversity. J. Microbiol. 52:120-128.
  13. Kim, S. H., M. J. Alam, M. J. Gu, K. W. Park, C. O. Jeon, J. K. Ha, K. K. Cho, and S. S. Lee. 2012. Effect of total mixed ration with fermented feed on ruminal in vitro fermentation, growth performance and blood characteristics of Hanwoo steers. Asian Australas. J. Anim. Sci. 25:213-223.
  14. Krizek, M., P. Kalac, and J. Peterka. 1993. Biogenic amines in silage. 3. The occurrence of six biogenic amines in farm-scale grass and maize silages. Arch Tierernahr 45:131-137.
  15. MacDonald, K. A., J. W. Penno, E. S. Kolver, W. A. C. Carter, and J. A. S. Lancaster. 1998. Balancing pasture and maize silage diets for dairy cows using urea, soybean meal or fishmeal. In Proceedings of the New Zealand Society of Animal Production. New Zealand Soc. Anim. Prod. 58:102-105.
  16. Mao, S. Y., G. Zhang, and W. Y. Zhu. 2008. Effect of disodium fumarate on ruminal metabolism and rumen bacterial communities as revealed by denaturing gradient gel electrophoresis analysis of 16S ribosomal DNA. Anim. Feed Sci. Tech. 140:293-306.
  17. Martin, S. A. and M. N. Streeter. 1995. Effect of malate on in vitro mixed ruminal microorganism fermentation. J. Anim. Sci. 73:2141-2145.
  18. Motoi, Y., Y. Obara, and K. Shimbayashi. 1984. Changes in histamine concentration of ruminal contents and plasma in cattle fed on a formula feed and rolled barley. Nihon Juigaku Zasshi 46:309-314.
  19. Muyzer, G. and K. Smalla. 1998. Application of denaturing gradient gel electrophoresis (DGGE) and temperature gradient gel electrophoresis (TGGE) in microbial ecology. Antonie van Leeuwenhoek 73:127-141.
  20. Plaizier, J. C., D. O. Krause, G. N. Gozho, and B. W. McBride. 2008. Subacute ruminal acidosis in dairy cows: The physiological causes, incidence and consequences. Vet. J. 176:21-31.
  21. Nübel, U., B. Engelen, A. Felske, J. Snaidr, A. Wieshuber, R. I. Amann, W. Ludwig, and H. Backhaus. 1996. Sequence heterogeneities of genes encoding 16S rRNAs in Paenibacillus polymyxa detected by temperature gradient gel electrophoresis. J. Bacteriol. 178:5636-5643.
  22. Offner, A., A. Bach, and D. Sauvant. 2003. Quantitative review of in situ starch degradation in the rumen. Anim. Feed Sci. Technol. 106:81-93.
  23. Phuntsok, T., M. A. Froetschel, H. E. Amos, M. Zheng, and Y. W. Huang. 1998. Biogenic amines in silage, apparent postruminal passage, and the relationship between biogenic amines and digestive function and intake by steers. J. Dairy Sci. 81:2193-2203.
  24. Russell, J. B. and R. L. Baldwin. 1979. Comparison of substrate affinities among several rumen bacteria: A possible determinant of rumen bacterial competition. Appl. Environ. Microbiol. 37:531-536.
  25. Russell, J. B. and P. J. Van Soest. 1984. In vitro ruminal fermentation of organic acids common in forage. Appl. Environ. Microbiol. 47:155-159.
  26. SAS. 2002. SAS/STAT. Statistical analysis systems for windows. Release 9.1. SAS Institute Inc., Cary, NC, USA.
  27. Seo, J. K., J. Yang, H. J. Kim, S. D. Upadhaya, W. M. Cho, and J. K. Ha. 2010. Effects of synchronization of carbohydrate and protein supply on ruminal fermentation, nitrogen metabolism and microbial protein synthesis in Holstein steers. Asian Australas. J. Anim. Sci. 23:1455-1461.
  28. Snyder, L. R., J. J. Kirkland, and J. L. Glajch. 1997. Practical HPLC Method Development. John Wiley & Sons, Inc. Hoboken, NJ, USA.
  29. Steidlova, S. and P. Kala. 2002. Levels of biogenic amines in maize silages. Anim. Feed Sci. Technol. 102:197-205.
  30. Tveit, B., F. Lingaas, M. Svendsen, and O. V. Sjaastad. 1992. Etiology of acetonemia in Norwegian cattle. 1. Effect of ketogenic silage, season, energy level, and genetic factors. J. Dairy Sci. 75:2421-2432.
  31. Stone, W. C. 2004. Nutritional approaches to minimize subacute ruminal acidosis and laminitis in dairy cattle. J. Dairy Sci. 87:E13-E26.
  32. Tabaru, H., E. Kadota, H. Yamada, N. Sasaki, and A. Takeuchi. 1988. Determination of volatile fatty acids and lactic acid in bovine plasma and ruminal fluid by high performance liquid chromatography. Jpn. J. Vet. Sci. 50:1124-1126.
  33. Thauer, R. K. 1998. Biochemistry of methanogenesis: A tribute to Marjory Stephenson: 1998 Marjory Stephenson Prize Lecture. Microbiology 144:2377-2406.
  34. van Beers-Schreurs, H. M. G., M. J. A. Nabuurs, L. Vellenga, H. J. K.-v. d. Valk, T. Wensing, and H. J. Breukink. 1998. Weaning and the weanling diet influence the villous height and crypt depth in the small intestine of pigs and alter the concentrations of short-chain fatty acids in the large intestine and blood. J. Nutr. 128:947-953.
  35. van Kessel, J. A. S. and J. B. Russell. 1996. The effect of pH on ruminal methanogenesis. FEMS Microbiol. Ecol. 20:205-210.
  36. Van Nevel, C. J. and D. I. Demeyer. 1977. Effect of monensin on rumen metabolism in vitro. Appl. Environ. Microbiol. 34:251-257.
  37. Van Os, M., J. P. Dulphy, and R. Baumont. 1995. The effect of protein degradation products in grass silages on feed intake and intake behaviour in sheep. Br. J. Nutr. 73:51-64.
  38. Walker, J. A. and D. L. Harmon. 1995. Influence of ruminal or abomasal starch hydrolysate infusion on pancreatic exocrine secretion and blood glucose and insulin concentrations in steers. J. Anim. Sci. 73(12):3766-3774.
  39. Williams, A. G. 1986. Rumen holotrich ciliate protozoa. Microbiol. Rev. 50:25-49.

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