Applied Microbiology and Biotechnology

, Volume 97, Issue 9, pp 4075–4081 | Cite as

Quantitative analysis of growth and volatile fatty acid production by the anaerobic ruminal bacterium Megasphaera elsdenii T81

Applied microbial and cell physiology


Megasphaera elsdenii T81 grew on either dl-lactate or d-glucose at similar rates (0.85 h−1) but displayed major differences in the fermentation of these substrates. Lactate was fermented at up to 210-mM concentration to yield acetic, propionic, butyric, and valeric acids. The bacterium was able to grow at much higher concentrations of d-glucose (500 mM), but never removed more than 80 mM of glucose from the medium, and nearly 60 % the glucose removed was sequestered as intracellular glycogen, with low yields of even-carbon acids (acetate, butyrate, caproate). In the presence of both substrates, glucose was not used until lactate was nearly exhausted, even by cells pregrown on glucose. Glucose-grown cultures maintained only low extracellular concentrations of acetate, and addition of exogenous acetate increased yields of butyrate, but not caproate. By contrast, exogenous acetate had little effect on lactate fermentation. At pH 6.6, growth rate was halved by exogenous addition of 60 mM propionate, 69 mM butyrate, 44 mM valerate, or 33 mM caproate; at pH 5.9, these values were reduced to 49, 49, 18, and 22 mM, respectively. The results are consistent with this species’ role as an effective ruminal lactate consumer and suggest that this organism may be useful for industrial production of volatile fatty acids from lactate if product tolerance could be improved. The poor fermentation of glucose and sensitivity to caproate suggests that this strain is not practical for industrial caproate production.


Lactate Glucose Megasphaera elsdenii Volatile fatty acids 



We thank C.L. Odt for the technical assistance and M.B. Hall for the useful suggestions regarding glycogen analysis. This research was supported by USDA-ARS CRIS project 3655-41000-06-00D.

Conflict of interest

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  1. Bradford MM (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254Google Scholar
  2. Gutierrez J, Davis RE, Lindahl IL, Warwick EJ (1959) Bacterial changes in the rumen during the onset of feed-lot bloat of cattle and characteristics of Peptostreptococcus elsdenii n. sp. Appl Microbiol 7:16–22Google Scholar
  3. Hall MB (2011) Isotrichid protozoa influence conversion of glucose to glycogen and other microbial products. J Dairy Sci 94:4589–4602CrossRefGoogle Scholar
  4. Hino T, Kuroda S (1993) Presence of lactate dehydrogenase and lactate racemase in Megasphaera elsdenii grown on glucose or lactate. Appl Environ Microbiol 59:255–259Google Scholar
  5. Hino T, Miyazaki K, Kuroda S (1991) Role of extracellular acetate in the fermentation of glucose by a ruminal bacterium, Megasphaera elsdenii. J Gen Appl Microbiol 37:121–129CrossRefGoogle Scholar
  6. Hino T, Shimada K, Maruyama T (1994) Substrate preference in a strain of Megasphaera elsdenii, a ruminal bacterium, and its implications in propionate production and growth competition. Appl Environ Microbiol 60:1827–1831Google Scholar
  7. Holtzapple M, Granda C (2009) Carboxylate platform: the MixAlco process part 1: comparison of three biomass conversion platforms. Appl Biochem Biotechnol 156:525–536CrossRefGoogle Scholar
  8. Kenealy WR, Waselefsky DM (1985) Studies on the substrate range of Clostridium kluyveri: the use of propanol and succinate. Arch Microbiol 141:187–194CrossRefGoogle Scholar
  9. Lange JP, Price R, Ayoub PM, Louis J, Petrus L, Clarke L, Gosselink H (2010) Valeric biofuels: a platform of cellulosic transportation fuels. Angew Chem Int Ed 49:4479–4483CrossRefGoogle Scholar
  10. Marounek M, Fliegrova K, Bartos S (1989) Metabolism and some characteristics of ruminal strains of Megasphaera elsdenii. Appl Environ Microbiol 55:1570–1573Google Scholar
  11. Meissner HH, Henning PH, Horn CH, Leeuw KL, Hagg FM, Fouche G (2010) Ruminal acidosis: a review with detailed reference to the controlling agent Megasphaera elsdenii NCIMB 41125. S Afr J Anim Sci 40:79–100Google Scholar
  12. Miller GL, Blum R, Glennon WE, Burton AL (1960) Measurement of carboxymethylcellulase activity. Anal Biochem 1:127–132CrossRefGoogle Scholar
  13. Pavlostathis SG, Miller TL, Wolin MJ (1988) Fermentation of insoluble cellulose by continuous cultures of Ruminococcus albus. Appl Environ Microbiol 54:2655–2659Google Scholar
  14. Piknová M, Bíres O, Javorsky P, Pristas P (2006) Limited genetic variability in Megasphaera elsdenii strains. Folia Microbiol (Praha) 51:299–302CrossRefGoogle Scholar
  15. Rogosa M (1972) Transfer of Peptostreptococcus elsdenii Gutierrez et al. to a new genus, Megasphaera (M. elsdenii (Gutierrez et al.) comb. nov.). Int J Syst Bacteriol 21:187–189CrossRefGoogle Scholar
  16. Russell JB, Baldwin LR (1979) Comparison of maintenance energy expenditures and growth yields among several rumen bacteria grown in continuous culture. Appl Environ Microbiol 37:537–543Google Scholar
  17. Russell JB, Hino T (1985) Regulation of lactate production in Streptococcus bovis: a spiraling effect that contributes to rumen acidosis. J Dairy Sci 68:1712–1721CrossRefGoogle Scholar
  18. Thauer RK, Jungermann K, Decker K (1977) Energy conservation in chemotrophic anaerobic bacteria. Bacteriol Rev 31:100–180Google Scholar
  19. Torii S, Tanaka H (2001) Carboxylic acids. In: Lund H, Hammerich O (eds) Organic electrochemistry, 4th edn. Marcel Dekker, New York, pp 499–535Google Scholar
  20. Wallace RJ, Chaudhary ME, McKain N, Walker ND (2004) Metabolic properties of Eubacterium pyruvativorans, a ruminal ‘hyper-ammonia-producing’ anaerobe with metabolic properties analogous to those of Clostridium kluyveri. Microbiology 150:2921–2930CrossRefGoogle Scholar
  21. Weimer PJ (2010) End product yields from the extraruminal fermentation of various polysaccharide, protein and nucleic acid components of biofuels feedstocks. Biores Technol 012:3254–3259Google Scholar
  22. Weimer PJ, Stevenson DM (2011) Isolation, characterization and quantification of Clostridium kluyveri from the bovine rumen. Appl Microbiol Biotechnol 94:461–466CrossRefGoogle Scholar
  23. Weimer PJ, Shi Y, Odt CL (1991) A segmented gas/liquid delivery system for continuous culture of microorganisms on solid substrates, and its use for growth of Ruminococcus flavefaciens on cellulose. Appl Microbiol Biotechnol 36:178–183CrossRefGoogle Scholar
  24. Wood WA (1961) Fermentation of carbohydrates and related compounds. In: Gunsalus IC, Stanier RY (eds) The bacteria, vol 2. Academic, New YorkGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2012 (outside the USA) 2012

Authors and Affiliations

  1. 1.US Dairy Forage Research Center, Agricultural Research ServiceUnited States Department of AgricultureMadisonUSA
  2. 2.Department of BacteriologyUniversity of Wisconsin-MadisonMadisonUSA

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