Dynamics of Polyphosphate-Accumulating Bacteria in Wastewater Treatment Plant Microbial Communities Detected via DAPI (4',6'-Diamidino-2-Phenylindole) and Tetracycline Labeling

• Published in: Applied and Environmental Microbiology Vol. 75; no. 7; pp. 2111 - 2121
• Main Authors: Günther, S, Trutnau, M, Kleinsteuber, S, Hause, G, Bley, T, Röske, I, Harms, H, Müller, S
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01.04.2009; American Society for Microbiology (ASM)
• Subjects: Bacteria; Bacteria - classification; Bacteria - isolation & purification; Bacteria - metabolism; bacterial communities; Biological and medical sciences; Biological treatment of waters; Biotechnology; Cations; Cell growth; Community involvement; cytoplasmic granules; Deoxyribonucleic acid; DNA; DNA, Bacterial - chemistry; DNA, Bacterial - genetics; DNA, Ribosomal - chemistry; DNA, Ribosomal - genetics; Environment and pollution; Environmental Microbiology; flow cytometry; Flow Cytometry - methods; Fluorescence; Fundamental and applied biological sciences. Psychology; Genes, rRNA; granules; Indoles - metabolism; Industrial applications and implications. Economical aspects; Microbial activity; Microbiology; Molecular Sequence Data; monitoring; phosphates; phosphorus; Phosphorus removal; Phylogeny; plant communities; Polyphosphates - metabolism; Pseudomonas; quantitative analysis; RNA, Bacterial - genetics; RNA, Ribosomal, 16S - genetics; Sequence Analysis, DNA; Sequence Homology, Nucleic Acid; Staining and Labeling; tetracycline; Tetracycline - metabolism; Tetrasphaera; wastewater; wastewater treatment; Wastewater treatment plants; Water Microbiology; Water Purification; Water treatment plants; Dynamics; Tagging; Bacteria; Waste water purification; Polyphosphates; Microbial community; Biological purification; Tetracycline
• ISSN: 0099-2240; 1098-5336; 1098-5336; 1098-6596
• DOI: 10.1128/aem.01540-08

Wastewater treatment plants with enhanced biological phosphorus removal represent a state-of-the-art technology. Nevertheless, the process of phosphate removal is prone to occasional failure. One reason is the lack of knowledge about the structure and function of the bacterial communities involved. Most of the bacteria are still not cultivable, and their functions during the wastewater treatment process are therefore unknown or subject of speculation. Here, flow cytometry was used to identify bacteria capable of polyphosphate accumulation within highly diverse communities. A novel fluorescent staining technique for the quantitative detection of polyphosphate granules on the cellular level was developed. It uses the bright green fluorescence of the antibiotic tetracycline when it complexes the divalent cations acting as a countercharge in polyphosphate granules. The dynamics of cellular DNA contents and cell sizes as growth indicators were determined in parallel to detect the most active polyphosphate-accumulating individuals/subcommunities and to determine their phylogenetic affiliation upon cell sorting. Phylotypes known as polyphosphate-accumulating organisms, such as a "Candidatus Accumulibacter"-like phylotype, were found, as well as members of the genera Pseudomonas and TETRASPHAERA: The new method allows fast and convenient monitoring of the growth and polyphosphate accumulation dynamics of not-yet-cultivated bacteria in wastewater bacterial communities.