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Development of a biofilm model for Listeria monocytogenes EGD-e

Abstract

The ability to form persistent biofilms makes the pathogenic bacterium Listeria monocytogenes a hazardous contaminant in food processing environments. Growth and biofilm formation of L. monocytogenes EGD-e were studied in defined medium (HTM) and in tryptic soy broth (TSB) with different supplements. TSB + 1% glucose gave optimal results. Using this medium, biofilm development on the model surface polystyrene (microtiter plate) was monitored by the standard crystal violet staining for adherent cells after bacterial cultivation for 24 and 48 h at five different temperatures (4, 18, 25, 30 and 37°C). In parallel, the matrix exopolysaccharide formed after 48 h of incubation was quantified by staining with ruthenium red. In both assays incubation at 30°C yielded the highest values. The formation of larger scale biofilms on dialysis membranes, placed on TSB agar with 1% glucose for 48 h, was studied by scanning electron microscopy. Contiguous and multilayered biofilms were observed at 18, 25, 30 and 37°C incubation temperature. The methodology is suitable for quantitative and microscopic studies and, in addition, yields sufficient cell mass for subsequent biochemical and molecular biological analyses.

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Acknowledgments

We thank Daniela Bunsen, University of Würzburg, for her technical assistance during SEM studies. This work was supported by grants to S.G. from the Department of Science and Technology, Govt. of India (SR/FT/L-89/2005) and from the Indian Council for Medical Research (5/8/9/56/2006-ECD-I) and to J.K. from the International Bureau/Bundesministerium für Bildung und Forschung, Germany (IND 06/010).

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Correspondence to Shubha Gopal.

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Zameer, F., Gopal, S., Krohne, G. et al. Development of a biofilm model for Listeria monocytogenes EGD-e. World J Microbiol Biotechnol 26, 1143–1147 (2010). https://doi.org/10.1007/s11274-009-0271-4

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Keywords

  • Biofilm
  • Cellulose membrane
  • Listeria monocytogenes
  • Polystyrene
  • Temperature
  • Scanning electron microscopy