This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Kuiper, M. W.
Right arrow Articles by van der Kooij, D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kuiper, M. W.
Right arrow Articles by van der Kooij, D.
Agricola
Right arrow Articles by Kuiper, M. W.
Right arrow Articles by van der Kooij, D.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, November 2004, p. 6826-6833, Vol. 70, No. 11
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.11.6826-6833.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Intracellular Proliferation of Legionella pneumophila in Hartmannella vermiformis in Aquatic Biofilms Grown on Plasticized Polyvinyl Chloride

Melanie W. Kuiper,1,2 Bart A. Wullings,3 Antoon D. L. Akkermans,2 Rijkelt R. Beumer,1 and Dick van der Kooij2,3*

Laboratories of Food Microbiology,1 Microbiology, Wageningen University and Research Center, Wageningen,2 Kiwa N.V. Water Research, Nieuwegein, The Netherlands3

Received 13 February 2004/ Accepted 25 June 2004

The need for protozoa for the proliferation of Legionella pneumophila in aquatic habitats is still not fully understood and is even questioned by some investigators. This study shows the in vivo growth of L. pneumophila in protozoa in aquatic biofilms developing at high concentrations on plasticized polyvinyl chloride in a batch system with autoclaved tap water. The inoculum, a mixed microbial community including indigenous L. pneumophila originating from a tap water system, was added in an unfiltered as well as filtered (cellulose nitrate, 3.0-µm pore size) state. Both the attached and suspended biomasses were examined for their total amounts of ATP, for culturable L. pneumophila, and for their concentrations of protozoa. L. pneumophila grew to high numbers (6.3 log CFU/cm2) only in flasks with an unfiltered inoculum. Filtration obviously removed the growth-supporting factor, but it did not affect biofilm formation, as determined by measuring ATP. Cultivation, direct counting, and 18S ribosomal DNA-targeted PCR with subsequent sequencing revealed the presence of Hartmannella vermiformis in all flasks in which L. pneumophila multiplied and also when cycloheximide had been added. Fluorescent in situ hybridization clearly demonstrated the intracellular growth of L. pneumophila in trophozoites of H. vermiformis, with 25.9% ± 10.5% of the trophozoites containing L. pneumophila on day 10 and >90% containing L. pneumophila on day 14. Calculations confirmed that intracellular growth was most likely the only way for L. pneumophila to proliferate within the biofilm. Higher biofilm concentrations, measured as amounts of ATP, gave higher L. pneumophila concentrations, and therefore the growth of L. pneumophila within engineered water systems can be limited by controlling biofilm formation.

* Corresponding author. Mailing address: Kiwa N.V. Water Research, Groningenhaven 7, P.O. Box 1072, 3430 BB Nieuwegein, The Netherlands. Phone: 31 (30) 6069634. Fax: 31 (30) 6061165. E-mail: Dick.van.der.Kooij{at}kiwa.nl.

Applied and Environmental Microbiology, November 2004, p. 6826-6833, Vol. 70, No. 11
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.11.6826-6833.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Valster, R. M., Wullings, B. A., Bakker, G., Smidt, H., van der Kooij, D. (2009). Free-Living Protozoa in Two Unchlorinated Drinking Water Supplies, Identified by Phylogenic Analysis of 18S rRNA Gene Sequences. Appl. Environ. Microbiol. 75: 4736-4746 [Abstract] [Full Text]  
  • Faulkner, G., Berk, S. G., Garduno, E., Ortiz-Jimenez, M. A., Garduno, R. A. (2008). Passage through Tetrahymena tropicalis Triggers a Rapid Morphological Differentiation in Legionella pneumophila. J. Bacteriol. 190: 7728-7738 [Abstract] [Full Text]  
  • Wang, W., Shor, L. M., LeBoeuf, E. J., Wikswo, J. P., Taghon, G. L., Kosson, D. S. (2008). Protozoan Migration in Bent Microfluidic Channels. Appl. Environ. Microbiol. 74: 1945-1949 [Abstract] [Full Text]  
  • Hindre, T., Bruggemann, H., Buchrieser, C., Hechard, Y. (2008). Transcriptional profiling of Legionella pneumophila biofilm cells and the influence of iron on biofilm formation. Microbiology 154: 30-41 [Abstract] [Full Text]  
  • Kuiper, M. W., Valster, R. M., Wullings, B. A., Boonstra, H., Smidt, H., van der Kooij, D. (2006). Quantitative Detection of the Free-Living Amoeba Hartmannella vermiformis in Surface Water by Using Real-Time PCR. Appl. Environ. Microbiol. 72: 5750-5756 [Abstract] [Full Text]  
  • Temmerman, R., Vervaeren, H., Noseda, B., Boon, N., Verstraete, W. (2006). Necrotrophic Growth of Legionella pneumophila.. Appl. Environ. Microbiol. 72: 4323-4328 [Abstract] [Full Text]  
  • Mampel, J., Spirig, T., Weber, S. S., Haagensen, J. A. J., Molin, S., Hilbi, H. (2006). Planktonic Replication Is Essential for Biofilm Formation by Legionella pneumophila in a Complex Medium under Static and Dynamic Flow Conditions. Appl. Environ. Microbiol. 72: 2885-2895 [Abstract] [Full Text]  
  • Borella, P., Montagna, M. T., Stampi, S., Stancanelli, G., Romano-Spica, V., Triassi, M., Marchesi, I., Bargellini, A., Tato, D., Napoli, C., Zanetti, F., Leoni, E., Moro, M., Scaltriti, S., Ribera D'Alcala, G., Santarpia, R., Boccia, S. (2005). Legionella Contamination in Hot Water of Italian Hotels. Appl. Environ. Microbiol. 71: 5805-5813 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»