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Changes to the rumen bacterial population of sheep with the addition of 2,4,6-trinitrotoluene to their diet

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Abstract

Previous work has shown that bacterial isolates from the sheep rumen are capable of detoxifying 2,4,6-trinitrotoluene (TNT) into polar constituents. In this study, the dietary effects of TNT on the sheep rumen microbial community were evaluated using molecular microbiology ecology tools. Rumen samples were collected from sheep fed with and without TNT added to their diet, genomic DNA was extracted, and the 16S rRNA-V3 gene marker was used to quantify changes in the microbial population in the rumen. Control and treatment samples yielded 533 sequences. Phylogenetic analyses were performed to determine the microbial changes between the two conditions. Results indicated the predominant bacterial populations present in the rumen were comprised of the phyla Firmicutes and Bacteroidetes, irrespective of presence/absence of TNT in the diet. Significant differences (P < 0.001) were found between the community structure of the bacteria under TNT (−) and TNT (+) diets. Examination of the TNT (+) diet showed an increase in the clones belonging to family Ruminococcaceae, which have previously been shown to degrade TNT in pure culture experiments.

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Abbreviations

TNT:

2,4,6-Trinitrotoluene

EPA:

Environmental protection agency

DGGE:

Denaturant gradient gel electrophoresis

LGCGPB:

Low GC Gram-positive bacteria

OTUs:

Operational taxonomic units

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Acknowledgments

This material is based upon research supported jointly funded by the Oregon Agricultural Experiment Station project ORE00871 and by the U.S. Department of Agriculture, under agreement nos. 58-6227-8-044 and 58-1265-6-076. Any opinions, findings, conclusion, or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect the view of the U.S. Department of Agriculture. The authors are indebted to D. J. Smith for obtaining the rumen samples used in the analysis. The authors would like to thank Ms. Zelda Zimmerman for editorial assistance and Katie Coleman for help with RDPII website analysis.

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Authors and Affiliations

  1. Department of Environmental and Molecular Toxicology, Oregon State University, Corvallis, OR, 97331, USA

    Sudeep Perumbakkam

  2. Department of Microbiology, Oregon State University, Corvallis, OR, 97331, USA

    Edward A. Mitchell

  3. Department of Veterinary Medicine, College of Veterinary Medicine, Oregon State University, 100 Magruder Hall, Corvallis, OR, 97331, USA

    A. Morrie Craig

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  1. Sudeep Perumbakkam
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  2. Edward A. Mitchell
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Correspondence to A. Morrie Craig.

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Perumbakkam, S., Mitchell, E.A. & Morrie Craig, A. Changes to the rumen bacterial population of sheep with the addition of 2,4,6-trinitrotoluene to their diet. Antonie van Leeuwenhoek 99, 231–240 (2011). https://doi.org/10.1007/s10482-010-9481-x

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