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Spontaneous bacterial cell lysis and biofilm formation in the colon of the Cape Dune mole-rat and the laboratory rabbit

  • Sanet H. Kotzé
  • Zoie E. Holzknecht
  • Anitra D. Thomas
  • Mary Lou Everett
  • Shanna Taylor
  • Larry D. Duckett
  • John Whitesides
  • Patrice McDermott
  • Shu S. Lin
  • William Parker
Methods and Protocols

Abstract

A wide range of techniques, including high-throughput DNA sequencing methods, have been applied to the evaluation of the normal intestinal flora. However, the inability to grow many of those species in culture imposes substantial constraints on the techniques used to evaluate this important community. The presence of biofilms in the normal gut adds further complexity to the issue. In this study, a flow cytometric analysis was used to separate intact bacterial cells, cell debris, and other particulate matter based on bacteria-specific staining and particle size. In addition, an analysis of biofilm formation using fluorescent light microscopy was conducted. Using these approaches, the ratio of bacterial cell debris to intact bacterial cells as a measure of spontaneous lysis of bacterial cells in the gut of the Cape dune mole-rat (Bathyergus suillus) and the laboratory rabbit (Oryctolagus cuniculus) was examined, and the degree of biofilm formation was semi-quantitatively assessed. The results suggest that the degree of spontaneous cell lysis was greater in the appendix than in the cecum in both the mole-rat and the rabbit. Further, the results point toward extensive epithelial-associated biofilm formation in the proximal mole-rat and rabbit large bowel, although the biofilms may be less structured than those found in laboratory rodents and in humans.

Keywords

Biofilm Cecal appendix Flow cytometry Mole-rat Rabbit Spontaneous lysis 

Notes

Acknowledgments

The authors would like to thank Dr J. M. O’Riain of the Zoology Department, UCT for kindly donating the mole-rat intestinal tract samples used in this study. This work was supported in part by awards AI-51445, P30 AI 64518, and U19 AI-678547 to the Duke Human Vaccine Institute Research Flow Cytometry Core Facility, and in part by the Fannie E. Rippel Foundation.

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Sanet H. Kotzé
    • 1
  • Zoie E. Holzknecht
    • 2
  • Anitra D. Thomas
    • 2
  • Mary Lou Everett
    • 2
  • Shanna Taylor
    • 2
  • Larry D. Duckett
    • 3
  • John Whitesides
    • 5
  • Patrice McDermott
    • 5
  • Shu S. Lin
    • 2
    • 4
  • William Parker
    • 2
  1. 1.Department of Biomedical SciencesFaculty of Health Sciences, University of StellenboschTygerbergSouth Africa
  2. 2.Department of SurgeryDuke University Medical CenterDurhamUSA
  3. 3.BD BiosciencesSan JoseUSA
  4. 4.Department of ImmunologyDuke University Medical CenterDurhamUSA
  5. 5.The Duke Human Vaccine Institute Flow Cytometry Core FacilityDuke University Medical CenterDurhamUSA

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