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Applied and Environmental Microbiology, July 2009, p. 4736-4746, Vol. 75, No. 14
0099-2240/09/$08.00+0     doi:10.1128/AEM.02629-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Free-Living Protozoa in Two Unchlorinated Drinking Water Supplies, Identified by Phylogenic Analysis of 18S rRNA Gene Sequences{triangledown} ,{dagger}

Rinske M. Valster,1,2* Bart A. Wullings,1 Geo Bakker,3 Hauke Smidt,2 and Dick van der Kooij1

KWR, Watercycle Research Institute, 3430 BB Nieuwegein, The Netherlands,1 Laboratory of Microbiology, Wageningen University, 6703 HB Wageningen, The Netherlands,2 Vitens Water Company, Leeuwarden, The Netherlands3

Received 17 November 2008/ Accepted 12 May 2009

Free-living protozoan communities in water supplies may include hosts for Legionella pneumophila and other undesired bacteria, as well as pathogens. This study aimed at identifying free-living protozoa in two unchlorinated groundwater supplies, using cultivation-independent molecular approaches. For this purpose, samples (<20°C) of treated water, distributed water, and distribution system biofilms were collected from supply A, with a low concentration of natural organic matter (NOM) (<0.5 ppm of C), and from supply B, with a high NOM concentration (7.9 ppm of C). Eukaryotic communities were studied using terminal restriction fragment length polymorphism and clone library analyses of partial 18S rRNA gene fragments and a Hartmannella vermiformis-specific quantitative PCR (qPCR). In both supplies, highly diverse eukaryotic communities were observed, including free-living protozoa, fungi, and metazoa. Sequences of protozoa clustered with Amoebozoa (10 operational taxonomic units [OTUs]), Cercozoa (39 OTUs), Choanozoa (26 OTUs), Ciliophora (29 OTUs), Euglenozoa (13 OTUs), Myzozoa (5 OTUs), and Stramenopiles (5 OTUs). A large variety of protozoa were present in both supplies, but the estimated values for protozoan richness did not differ significantly. H. vermiformis was observed in both supplies but was not a predominant protozoan. One OTU with the highest similarity to Acanthamoeba polyphaga, an opportunistic human pathogen and a host for undesired bacteria, was observed in supply A. The high level of NOM in supply B corresponded with an elevated level of active biomass and with elevated concentrations of H. vermiformis in distributed water. Hence, the application of qPCR may be promising in elucidating the relationship between drinking water quality and the presence of specific protozoa.

* Corresponding author. Mailing address: KWR, Watercycle Research Institute, Groningenhaven 7, P.O. Box 1072, 3430 BB Nieuwegein, The Netherlands. Phone: 31 30 606 9651. Fax: 31 30 606 1165. E-mail: rinske.valster{at}kwrwater.nl

{triangledown} Published ahead of print on 22 May 2009.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, July 2009, p. 4736-4746, Vol. 75, No. 14
0099-2240/09/$08.00+0     doi:10.1128/AEM.02629-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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