Legionella pp 79-89 | Cite as

Migration of Acanthamoeba castellanii Through Legionella Biofilms

  • Ramon Hochstrasser
  • Hubert HilbiEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1921)


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.

Key words

Acanthamoeba castellanii Biofilm formation Cell motility Confocal microscopy Green fluorescent protein Host-pathogen interactions Legionella pneumophila Single-cell tracking Type IV secretion 



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


ACES yeast extract




Confocal laser scanning microscope


Charcoal yeast extract


Green fluorescent protein


Optical density at 600 nm


Peptone yeast extract glucose



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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Institute of Medical MicrobiologyUniversity of ZürichZürichSwitzerland

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