Combining flow cytometry and 16S rRNA gene pyrosequencing: A promising approach for drinking water monitoring and characterization
AuthorsPrest, Emmanuelle I E C
El Chakhtoura, Joline
Hammes, Frederik A.
van Loosdrecht, Mark C.M.
Vrouwenvelder, Johannes S.
KAUST DepartmentBiological and Environmental Sciences and Engineering (BESE) Division
Water Desalination and Reuse Research Center (WDRC)
Environmental Science and Engineering Program
Environmental Biotechnology Research Group
Environmental Science and Engineering
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AbstractThe combination of flow cytometry (FCM) and 16S rRNA gene pyrosequencing data was investigated for the purpose of monitoring and characterizing microbial changes in drinking water distribution systems. High frequency sampling (5min intervals for 1h) was performed at the outlet of a treatment plant and at one location in the full-scale distribution network. In total, 52 bulk water samples were analysed with FCM, pyrosequencing and conventional methods (adenosine-triphosphate, ATP; heterotrophic plate count, HPC). FCM and pyrosequencing results individually showed that changes in the microbial community occurred in the water distribution system, which was not detected with conventional monitoring. FCM data showed an increase in the total bacterial cell concentrations (from 345±15×103 to 425±35×103cellsmL-1) and in the percentage of intact bacterial cells (from 39±3.5% to 53±4.4%) during water distribution. This shift was also observed in the FCM fluorescence fingerprints, which are characteristic of each water sample. A similar shift was detected in the microbial community composition as characterized with pyrosequencing, showing that FCM and genetic fingerprints are congruent. FCM and pyrosequencing data were subsequently combined for the calculation of cell concentration changes for each bacterial phylum. The results revealed an increase in cell concentrations of specific bacterial phyla (e.g., Proteobacteria), along with a decrease in other phyla (e.g., Actinobacteria), which could not be concluded from the two methods individually. The combination of FCM and pyrosequencing methods is a promising approach for future drinking water quality monitoring and for advanced studies on drinking water distribution pipeline ecology. © 2014 Elsevier Ltd.
SponsorsThe studies presented in this article were supported by funds from Evides Waterbedrijf and King Abdullah University of Science and Technology. The authors like to specifically thank Rinnert Schurer, Jan Bahlman and Elodie Loubineaud for their technical support.
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