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Applied and Environmental Microbiology, November 2009, p. 7115-7124, Vol. 75, No. 22
0099-2240/09/$08.00+0     doi:10.1128/AEM.00739-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Rumen Microbiome Composition Determined Using Two Nutritional Models of Subacute Ruminal Acidosis{triangledown}

Ehsan Khafipour,1,{dagger} Shucong Li,1,{dagger} Jan C. Plaizier,1 and Denis O. Krause1,2*

Department of Animal Science, University of Manitoba, Winnipeg, Manitoba, Canada,1 Department of Medical Microbiology, University of Manitoba, Winnipeg, Manitoba, Canada2

Received 2 April 2009/ Accepted 18 September 2009

Subacute ruminal acidosis (SARA) is a metabolic disease in dairy cattle that occurs during early and mid-lactation and has traditionally been characterized by low rumen pH, but lactic acid does not accumulate as in acute lactic acid acidosis. It is hypothesized that factors such as increased gut permeability, bacterial lipopolysaccharides, and inflammatory responses may have a role in the etiology of SARA. However, little is known about the nature of the rumen microbiome during SARA. In this study, we analyzed the microbiome of 64 rumen samples taken from eight lactating Holstein dairy cattle using terminal restriction fragment length polymorphisms (TRFLP) of 16S rRNA genes and real-time PCR. We used rumen samples from two published experiments in which SARA had been induced with either grain or alfalfa pellets. The results of TRFLP analysis indicated that the most predominant shift during SARA was a decline in gram-negative Bacteroidetes organisms. However, the proportion of Bacteroidetes organisms was greater in alfalfa pellet-induced SARA than in mild or severe grain-induced SARA (35.4% versus 26.0% and 16.6%, respectively). This shift was also evident from the real-time PCR data for Prevotella albensis, Prevotella brevis, and Prevotella ruminicola, which are members of the Bacteroidetes. The real-time PCR data also indicated that severe grain-induced SARA was dominated by Streptococcus bovis and Escherichia coli, whereas mild grain-induced SARA was dominated by Megasphaera elsdenii and alfalfa pellet-induced SARA was dominated by P. albensis. Using discriminant analysis, the severity of SARA and degree of inflammation were highly correlated with the abundance of E. coli and not with lipopolysaccharide in the rumen. We thus suspect that E. coli may be a contributing factor in disease onset.

* Corresponding author. Mailing address: Department of Animal Science, University of Manitoba, Winnipeg, MB, Canada R3T 2N2. Phone: (204) 474-6126. Fax: (204) 474-7628. E-mail: denis_krause{at}umanitoba.ca

{triangledown} Published ahead of print on 25 September 2009.

{dagger} These authors contributed equally to the research.

Applied and Environmental Microbiology, November 2009, p. 7115-7124, Vol. 75, No. 22
0099-2240/09/$08.00+0     doi:10.1128/AEM.00739-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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