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Legionella pp 79–89Cite as

Migration of Acanthamoeba castellanii Through Legionella Biofilms

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1921))

Abstract

The amoeba-resistant bacterium Legionella pneumophila infects humans through aerosols and thereby can cause a life-threatening pneumonia termed Legionnaires’ disease. In the environment L. pneumophila forms and colonizes biofilms, which usually comprise complex multispecies communities. In these biofilms L. pneumophila persists and replicates intracellularly in protozoa, such as the amoeba Acanthamoeba castellanii. The interactions between sessile L. pneumophila in biofilms and their natural protozoan hosts are not understood on a molecular level. Here, we describe a method to visualize by confocal microscopy the formation and architecture of mono-species L. pneumophila biofilms. Furthermore, we describe and quantify the migration or “grazing” of A. castellanii in the biofilm. This allows investigating on a molecular and cellular level L. pneumophila biofilm formation and Legionella-amoeba interactions within biofilms.

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Abbreviations

ACES:

N-(2-acetamido)-2-aminoethanesulfonic acid

AYE:

ACES yeast extract

Cam:

Chloramphenicol

CLSM:

Confocal laser scanning microscope

CYE:

Charcoal yeast extract

GFP:

Green fluorescent protein

OD600:

Optical density at 600 nm

PYG:

Peptone yeast extract glucose

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Acknowledgments

This work was supported by the University of Zürich (“Forschungskredit Candoc”; K-42226-01-01) and the Swiss National Science Foundation (SNF; 31003A_153200).

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Authors and Affiliations

  1. Institute of Medical Microbiology, University of Zürich, Zürich, Switzerland

    Ramon Hochstrasser & Hubert Hilbi

Authors
  1. Ramon Hochstrasser
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  2. Hubert Hilbi
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Corresponding author

Correspondence to Hubert Hilbi .

Editor information

Editors and Affiliations

  1. Institut Pasteur, Biologie des Bactéries Intracellulaires, Paris, France

    Carmen Buchrieser

  2. Institute of Medical Microbiology, University of Zürich, Zürich, Switzerland

    Hubert Hilbi

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Hochstrasser, R., Hilbi, H. (2019). Migration of Acanthamoeba castellanii Through Legionella Biofilms. In: Buchrieser, C., Hilbi, H. (eds) Legionella. Methods in Molecular Biology, vol 1921. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9048-1_5

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  • DOI: https://doi.org/10.1007/978-1-4939-9048-1_5

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9047-4

  • Online ISBN: 978-1-4939-9048-1

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