Starved viable but non-culturable (VBNC) Legionella strains can infect and replicate in amoebae and human macrophages

Elisabeth Dietersdorfer, Alexander Kirschner*, Barbara Schrammel, Anna Ohradanova-Repic, Hannes Stockinger, Regina Sommer, Julia Walochnik, Sílvia Cervero-Aragó

*Corresponding author for this work

Research output: Journal article (peer-reviewed)Journal article

59 Citations (Scopus)


Legionella infections are among the most important waterborne infections with constantly increasing numbers of cases in industrialized countries, as a result of aging populations, rising numbers of immunocompromised individuals and increased need for conditioned water due to climate change. Surveillance of water systems is based on microbiological culture-based techniques; however, it has been shown that high percentages of the Legionella populations in water systems are not culturable. In the past two decades, the relevance of such viable but non-culturable (VBNC) legionellae has been controversially discussed, and whether VBNC legionellae can directly infect human macrophages, the primary targets of Legionella infections, remains unclear. In this study, it was demonstrated for the first time that several starved VBNC Legionella strains (four L. pneumophila serogroup 1 strains, a serogroup 6 strain and a L. micdadei strain) can directly infect different types of human macrophages and amoebae even after one year of starvation in ultrapure water. However, under these conditions, the strains caused infection with reduced efficacy, as represented by the lower percentages of infected cells, prolonged time in co-culture and higher multiplicities of infection required. Interestingly, the VBNC cells remained mostly non-culturable even after multiplication within the host cells. Amoebal infection by starved VBNC Legionella, which likely occurs in oligotrophic biofilms, would result in an increase in the bacterial concentration in drinking-water systems. If cells remain in the VBNC state, the real number of active legionellae will be underestimated by the use of culture-based standard techniques. Thus, further quantitative research is needed in order to determine, whether and how many starved VBNC Legionella cells are able to cause disease in humans.

Original languageEnglish
Pages (from-to)428-438
Number of pages11
JournalWater Research
Publication statusPublished - 15 Sept 2018
Externally publishedYes


  • Acanthamoeba
  • Legionella
  • Macrophages
  • Starvation
  • VBNC

ASJC Scopus subject areas

  • Environmental Engineering
  • Civil and Structural Engineering
  • Ecological Modeling
  • Water Science and Technology
  • Waste Management and Disposal
  • Pollution


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