Role of "G-bacteria" in anaerobic substrate uptake in a SBR with no phosphorus removal

La Trobe University Faculty of Science, Technology and Engineering Murray Darling Freshwater Research Centre

MDFRC item.

Biomass from an SBR running with no enhanced biological phosphorus removal (EBPR) but which exhibited anaerobic assimilation of glucose and acetate, was dominated by "G-bacteria", cocci in tetrads and clusters. Extracted 16S rDNA was amplified by PCR and then analysed using Denaturing Gradient Gel Electrophoresis (DGGE). Major bands were extracted and their sequences determined. Clone libraries were also prepared, the 16S rDNA extracted, PCR performed and the resultant fragments run by DGGE to aid in identifying the DGGE bands and provide fuller sequences than available by DGGE alone. The two approaches together allowed several bands to be identified. Probes for FISH analyses were designed for some of these in attempts to see to which phylogenetic group "G-bacteria" belonged, and whether they represented the dominant bands detected by DGGE. Then FISH/Microautoradiography (MAR) was used in attempts to see which bacteria there were assimilating substrates anaerobically. Results indicated that the "G-bacteria" were phylogenetically diverse, but mainly alpha-proteobacteria and members of the high G+C% Gram-positive bacteria. Not all of these could assimilate glucose and/or acetate anaerobically, and Amaricoccus, the original "G-bacteria" of Cech and Hartman, was not detected.