ANALYSIS OF MICROBIAL COMMUNITY STRUCTURE AND IN SITU ACTIVITY OF NITRIFYING BIOFILMS

Abstract

Wastewater biofilms are very complex multispecies biofilms, displaying considerable heterogeneity with respect to both the microorganisms present and their physicochemical microenvironments. To understand the eco-physiology of individual microorganisms in the biofilm, techniques and tools with a high spatial and temporal resolution are required for direct detection of the spatial distributions of microbial species and their activities in minimally disturbed their natural habitats (e.g., biofilms). In this paper, we will, therefore, address the great potential of the combined use of the current FISH technique and microelectrodes to study the microbial ecology of complex microbial communities such as biofilms. The combination of these two techniques will provide reliable and direct information about relationships between in situ microbial activity and the occurrence of specific microorganisms in biofilms. As an example of the combined study, we will illustrate the in situ spatial organization of ammonia-oxidizing and nitrite-oxidizing bacteria on fine scale in autotrophic nitrifying biofilms by applying the full-cycle of 16S rRNA approach followed by fluorescence in situ hybridization (FISH), which is linked to their in situ activity distributions at a similar resolution determined by use of microelectrodes. The combination of these techniques allows relating in situ microbial activity directly to occurrence of nitrifying bacteria population.