Applied Microbiology and Biotechnology

, Volume 71, Issue 3, pp 329–338 | Cite as

Use of real time PCR to determine population profiles of individual species of lactic acid bacteria in alfalfa silage and stored corn stover

  • David M. Stevenson
  • Richard E. Muck
  • Kevin J. Shinners
  • Paul J. Weimer
Applied microbial and cell physiology
First Online:
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Accepted:

Abstract

Real-time polymerase chain reaction (RT-PCR) was used to quantify seven species of lactic acid bacteria (LAB) in alfalfa silage prepared in the presence or absence of four commercial inoculants and in uninoculated corn stover harvested and stored under a variety of field conditions. Species-specific PCR primers were designed based on recA gene sequences. Commercial inoculants improved the quality of alfalfa silage, but species corresponding to those in the inoculants displayed variations in persistence over the next 96 h. Lactobacillus brevis was the most abundant LAB (12 to 32% of total sample DNA) in all of the alfalfa silages by 96 h. Modest populations (up to 10%) of Lactobacillus plantarum were also observed in inoculated silages. Pediococcus pentosaceus populations increased over time but did not exceed 2% of the total. Small populations (0.1 to 1%) of Lactobacillus buchneri and Lactococcus lactis were observed in all silages, while Lactobacillus pentosus and Enterococcus faecium were near or below detection limits. Corn stover generally displayed higher populations of L. plantarum and L. brevis and lower populations of other LAB species. The data illustrate the utility of RT-PCR for quantifying individual species of LAB in conserved forages prepared under a wide variety of conditions.

Keywords

Lactobacillus Lactic Acid Bacterium Corn Stover Lactic Acid Bacterium Species Commercial Inoculant 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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Notes

Acknowledgements

This research was supported by the Agricultural Research Service, US Department of Agriculture, CRIS projects 3655-41000-004-00D and 3655-31000-018-00D. We thank Dave Spangler (Agri-King, Inc.) for providing cultures. We also thank Alltech, Inc., Chr. Hansen Biosystems, Genus Breeding Ltd., and Pioneer Hi-Bred International for providing bacterial inoculant products.

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Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • David M. Stevenson
    • 1
  • Richard E. Muck
    • 1
    • 2
    • 4
  • Kevin J. Shinners
    • 1
  • Paul J. Weimer
    • 2
    • 3
  1. 1.Department of Biological Systems EngineeringUniversity of Wisconsin-MadisonMadisonUSA
  2. 2.United States Dairy Forage Research Center, Agricultural Research ServiceUS Department of AgricultureMadisonUSA
  3. 3.Department of BacteriologyUniversity of Wisconsin-MadisonMadisonUSA
  4. 4.USDA-ARS-USDFRCMadisonUSA

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