Identity, abundance and ecophysiology of filamentous bacteria belonging to the Bacteroidetes present in activated sludge plants

1 Department of Biotechnology, Chemistry and Environmental Engineering, Aalborg University, Sohngaardsholmsvej 57, DK-9000 Aalborg, Denmark 2 CNR, Water Research Institute, Via Reno 1, 00198 Rome, Italy 3 Grontmij, De Holle Bilt 22, 3732 HM De Bilt Postbus 203, 3730 AE De Bilt, The Netherlands 4 VERMICON AG, Emmy-Noether-Str. 2, 80992 Munich, Germany 5 ASIS vof, Deventerweg 38, 7203 AK Zutphen, The Netherlands 6 BIOCLEAR Environmental Biotechnology, Rozenburglaan 13, 9727 DL Groningen, The Netherlands Correspondence Per Halkjær Nielsen phn{at}bio.aau.dk Filamentous members of the Bacteroidetes are commonly observed in activated sludge samples originating from both municipal and industrial wastewater treatment plants (WWTP), where they occasionally can cause bulking. Several oligonucleotide 16S rRNA-targeted probes were designed to target filaments with a needle-like appearance similar to Haliscomenobacter hydrossis . The design of these probes was based on an isolate and a sequence obtained from a micromanipulated filament. The abundance of filamentous Bacteroidetes was determined in 126 industrial samples applying already published and the newly developed probes. Small populations were found in 62 % of the WWTP investigated. However, only relatively few WWTP (13 %) contained large populations of filamentous Bacteroidetes potentially responsible for bulking incidences. The identity of the most abundant filamentous Bacteroidetes with H. hydrossis morphology could be detected by probes CFB719, SAP-309 and the newly designed probe HHY-654. A comprehensive study on the ecophysiology of probe-defined Bacteroidetes populations was conducted on Danish and Czech samples. The studies revealed that they were specialized bacteria involved in degradation of sugars, e.g. glucose and N -acetylglucosamine, and may participate in the conversion of lipopolysaccharides and peptidoglycan liberated by decaying cells. Many surface-associated exo-enzymes were excreted, e.g. chitinase, glucuronidase, esterase and phosphatase, supporting conversion of polysaccharides and possibly other released cell components. The role of filamentous bacteria with a H. hydrossis -like morphology in the activated sludge ecosystem is discussed. Abbreviations: ELF, enzyme-labelled fluorescence; FI, filament index; FISH, fluorescence in situ hybridization; MAC, microsphere adhesion to cells; MAR, microautoradiography; WWTP, wastewater treatment plant(s) The GenBank/EMBL/DDBJ accession numbers for th