Phylogenetic analysis and in situ identification of “Nostocoida limicola”-like filamentous bacteria in activated sludge from industrial wastewater treatment plants

Abstract

The diversity of filamentous bacteria present in industrial wastewater treatment plants was analysed by a combination of classical and molecular-biological approaches. Many unknown filamentous bacteria were observed in about 80 screened activated sludge samples from different industries with sometimes severe bulking sludge problems. A special focus was paid to filaments which resembled "Nostocoida limicola", a filamentous bacterium which was found to be present in many WWTPs. These filamentous bacteria are hardly cultivable and only one strain was obtained and maintained in co-culture with a yeast. The 16S rRNA sequences of several other "Nostocoida limicola"-like filamentous bacteria from different sludge samples were obtained by micromanipulation and different molecular-biological methods. The sequences were phylogenetically analyzed and specific molecular probes were developed and applied. The results clearly demonstrate that "Nostocoida limicola"-like filaments from industrial WWTPs are different from all other "Nostocoida limicola" types investigated so far. Our strains are affiliated to the alpha-subclass of Proteobacteria.

Meganema perideroedes gen. nov., sp. nov., a filamentous alphaproteobacterium from activated sludge

“…Thus, the isolated strains look like the Gram-variable filamentous bacteria in activated sludge with the 'Nostocoida limicola' type II morphology described by Eikelboom & van Buijsen (1983). This morphotype was also observed in previous studies (Snaidr et al, 2002;Schade et al, 2002;Blackall et al, 2000;Levantesi et al, 2004). Our results confirm the observation that different filaments from the 'N.…”

Design and application of oligonucleotide probes for fluorescent in situ identification of the filamentous bacterial morphotype Nostocoida limicola in activated sludge

“…A similar observation was made with N. limicola II, in which filaments unambiguously identified microscopically as this morphotype responded not only to the NLIMII and HGC 69a probes, as expected from the sequence data from cultured isolates of this bacterium , but also to the ALF 1b and ALF 968 probes for the a-Proteobacteria (Fig. N. limicola II filaments belonging to both the a-Proteobacteria (Snaidr et al, 2001) and the Chloroflexus group (Schade et al, 2001) have recently been described based on other cultured isolates, illustrating how phylogenetically diverse this morphotype is, and 16S rRNA targeted FISH probes are available for their in situ identification in activated sludge plants. In fact the a-Proteobacterial N. limicola II were often the dominant filament in some biomass samples examined, and both phylogenetic types were frequently seen (Table 2) in biomass samples from a single plant.…”

Ecophysiology of the Actinobacteria in activated sludge systems

“…Yet some data with the actinobacterial member of the polyphyletic 'Nostocoida limicola' morphotype Snaidr et al 2002), seen often in bulking and foaming plants around the world suggest it may possess such as capability. In most published plant survey data no distinction is made between the different 'N.…”