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http://dx.doi.org/10.5187/JAST.2005.47.4.615

Study on Rumen Cellulolytic Bacterial Attachment and Fermentation Dependent on Initial pH by cPCR  

Kim, M.S. (School of Agricultural Biotechnology, Seoul National University)
Sung, H.G. (School of Agricultural Biotechnology, Seoul National University)
Kim, H.J. (School of Agricultural Biotechnology, Seoul National University)
Lee, Sang-S. (Research Institute, Genebiotech Co.)
Chang, J.S. (Department of Agricultural Science, Korea National Open University)
Ha, J.K. (School of Agricultural Biotechnology, Seoul National University)
Publication Information
Journal of Animal Science and Technology / v.47, no.4, 2005 , pp. 615-624 More about this Journal
Abstract
The cPCR technique was used to monitor rumen fermentation and attachment of Fibrobacter succinogenes to cellulose at different pH in the in vitro culture medium. The target fragments of 16S rDNA(445 bp) were amplified from genomic DNA of F. succinogenes with specific primers and internal controls(205 bp) were constructed. Cell counts were estimated from the amounts of genomic DNA, which was calculated from cPCR results. F. succinogenes in pH 6.8 and 6.2 showed apparently higher attachment than in pH 5.8 during all incubation time. There were some difference between pH 6.8 and 6.2 in the degree of attachment, but the different was not significant (P>0.05). Cellulose degradation increased in process of incubation time and the increasing rate was higher when initial pH was higher. The pH in culture medium decreased regardless of initial pH in course of incubation time. After 24 h of incubation, medium pH was dropped by 0.24, 0.58 and 0.16 units from original medium pH 6.8, 6.2 and 5.8, respectively. More gas was produced at higher initial pH in the same manner as in cellulose degradation. In summery, Initial pH of rumen culture in vitro significantly influenced cellulose digestion, gas production, pH change and bacterial attachment. Especially, low pH(5.8) resulted in much lower bacterial attachment and fiber digestion compared to higher medium pH.
Keywords
Initial medium pH; F. succinogenes; Attachment; Rumen fermentation; cPCR;
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1 Dehority, B. A., Tirabasso, P. A. and Grifo, A. P. Jr. 1989. Most-probable-number procedures for enumerating ruminal bacteria, including in the simultaneous estimation of total and cellulolytic numbers in one medium. Appl. Environ. Microbiol. 55:2789-2792
2 Duncan, D. B. 1955. Multiple range and multiple F test. Biometerics. 11:1   DOI   ScienceOn
3 Firsberg, D. W., Cheng, K. J. and White, B. A. 1997. Polysaccharide degradation in the rumen and large intestine. In Gastrointestinal Microbiology, R. I. Mackie aqnd B. A. White(Eds), Chapman and Hall, New york, U.S.A, p319-379
4 Grant, R. J. 1994. Influence of com and sorghum starch on the in vitro kinetics of forage fiber digestion. J. Dairy Sci. 77:1563-1569   DOI
5 Mould, F. L., Orskov, E. R. and Mann, S. O. 1984. Associative effects of mixed feeds. I. effects of type and level of supplementation and the influence of the rumen fluid pH on cellulolysis in vivo and dry matter digestion of various roughages. Anim. Feed Sci. Technol. 10:15-30   DOI   ScienceOn
6 Ogimoto, K. and Imai, S. 1981 Atlas of Rumen Microbiology, Japan Scientific Societies Press, Tokyo
7 Roger, V., Fonty, G., Komisarczuk-Bony, S. and Gouet, P. 1990. Effects of physicochemical factors on the adhesion to cellulose avicel of the ruminal bacteria Ruminococcus jlavefaciens and Fibrobacter succinogenes subsp. succinogenes. Appl. Environ. Microbiol. 56:3081
8 Owens, F. N., Secrist, D. S., Hill, W. J. and Gill, D. R. 1998. Acidosis in cattle: a review. J. Anim, Sci. 76:275-286
9 Reilly, K. and Attwood, G. T. 1998. Detection of Clostridium proteoclasticum and closely related strains in the rumen by competitive PCR. Appl. Environ. Microbiol. 64:907-913
10 Mourino, F., Akkarawongsa, R. and Weimer, P. J. 2001. Initial pH as a determinant of cellulose digestion rate by mixed ruminal microorganisms in vitro. J. Dairy Sci. 84:848-859   DOI   ScienceOn
11 Ogata, K., Aminov, R. I., Nagamine, T., Sugiura, M., Tajima, K., Mitsumori, M., Sekizaki, T., Kudo, H., Minato, H. and Benno, Y. 1997. Construction of a Fibrobacter succinogenes Genomic Map and Demonstration of Diversity at the Genomic Level. Curr. Microbiol. 35:22-27   DOI   ScienceOn
12 Bae, H. D., McAllister, T. A., Kokko, E. G., Leggett, F. L., Yanke, L. J.. Jakober, K. D., Ha J. K., Shin, H. T. and Cheng, K. J. 1997. Effect of silica on the colonization of rice straw by ruminal bacteria. Anim. Feed Sci. Technol. 65: 165-181   DOI   ScienceOn
13 Koike, S., Pan, J., Kobayashi, Y. and Tanaka, K. 2003. Kinestics of in sacco fiber-attachment of representative ruminal cellulolytic bacteria monitored by competitive PCR. J. Dairy Sci. 86: 1429-1435   DOI   ScienceOn
14 Grant, R. J. and Weidner, S. J. 1992. Digestion kinetics of fiber: influence of in vitro buffer pH varied within observed physiological range. J. Dairy Sci. 75:1060-1068   DOI
15 Hungate, R. E. 1966. The Rumen and Its Microbes. Academic Press. New York, N. Y
16 Koike, S. and Kobayashi, Y. 2001. Development and use of competitive PCR assays for the rumen cellulolytic bacteria: Fibrobacter succinogenes, Ruminococcus albus and Ruminococcus jlavefaciens. FEMS Microbiol. Lett. 204: 361-366   DOI   ScienceOn
17 McAllister, T. A., Bae, H. D., Jones, G. A. and Cheng, K. J. 1994. Microbial attachment and feed digestion in the rumen. J. Anim. Sci. 72:3004-3018
18 McDougall, E. I. 1948. Studies on ruminant saliva I. The composition and output of sheep's saliva. Biochem. J. 43:99-109   DOI
19 Miron, J., Ben-Ghedalia, D. and Morrisont, M. 2001. Adhesion mechanism of rumen cellulolytic bacteria. J. dairy Sci. 84:1294-1309   DOI   ScienceOn
20 Reilly, K., Carruthers, V. R and Attwood, G. T. 2002. Design and use of 16S ribosomal DNA-directed primers in competitive PCRs to enumerate proteolytic bacteria in the rumen. 43:259-270   DOI   ScienceOn
21 Russell, J. B. and Rychlik, J. L. 2001. Factors that alter rumen microbial ecology. Science. 292:1119-1122   DOI   ScienceOn
22 Russell, J. B. and Wilson, D. B. 1996. Why are ruminal cellulolutic bacteria unable to digest cellulose at low pH? J. Dairy Sci. 70: 1503-1509
23 SAS. 1996. User's Guide. Version 6.12. Statistical Analysis System Inst. Inc. Cary NC. USA
24 Slyter, L. L. 1976. Influence of acidosis on rumen function. J. Anim. Sci. 43:910-929   DOI
25 Stahl. D. A.. Flesher. B.. Mansfield. H. R. and Montgomery. L. 1988. Use of phylogenetically based hybridization probes for studies of ruminal microbial ecology. Appl. Environ. Microbiol. 54:1079-1084
26 Woese. C. R. 1987. Bacterial evolution. Microbiol. Rev. 51 :221-227
27 Woese. C. R. 1994. There must be a prokaryote somewhere: microbiology's search for itself. Microbiol Rev. 58: 1-9
28 하종규, 이성실, 문양수, 김창현. 2005. 반추동물 영양생리학. 서울대학교출판부. pp. 122-249