Microbial diversity is generally considered as having no effect on the major processes of the ecosystem such as respiration or nutrient assimilation. However, information about the impact of diversity on minor functions such as xenobiotic degradation is scant. We studied the role of diversity on the capacity of an activated-sludge microbial community to eliminate phenanthrene, a polycyclic aromatic hydrocarbon. We also assessed the impact of diversity erosion on the ability of activated sludge to oxidize a wide range of organic compounds. The diversity of activated sludge was artificially modified by dilution to extinction followed by regrowth stage which led to communities with similar biomass but displaying a diversity gradient. The capacity of activated-sludge community to degrade phenanthrene was greatly modified: at high levels of diversity, the community was able to mineralize phenanthrene whereas at medium levels it first of all partially lost its ability to mineralize this pollutant and at the lowest diversity, the activated sludge completely lost its capacity to transform phenanthrene. Diversity depletion also reduced the metabolic diversity and biomass productivity of sewage-activated sludge. This study demonstrates that diversity erosion can greatly affect major ecosystem services such as pollutant removal.

Efficient dissociation of microorganisms from their aggregate matrix is required to study the microorganisms without interaction with their native environment (e.g., biofilms, flocs, granules, etc.) and to assess their community composition through the application of molecular or microscopy techniques. To this end, we combined enzymatic treatments and a cell extraction by density gradient to efficiently recover anaerobic microorganisms from urban wastewater treatment plant sludge. The enzymes employed (amylase, cellulase, DNase, and pectinase) as a pretreatment softly disintegrated the extrapolymeric substances (EPS) interlocked with the microorganisms. The potential damaging effects of the applied procedure on bacterial and archaeal communities were assessed by studying the variations in density (using quantitative PCR), diversity (using capillary electrophoresis single-strand conformation polymorphism fingerprinting [CE-SSCP]), and activity (using a standard anaerobic activity test) of the extracted microorganisms. The protocol preserved the general capacity of the microbial community to produce methane under anaerobic conditions and its diversity; particularly the archaeal community was not affected in terms of either density or structure. This cell extraction procedure from the matrix materials offers interesting perspectives for metabolic, microscopic, and molecular assays of microbial communities present in complex matrices constituted by bioaggregates or biofilms.

Urban sludge produced on wastewater treatment plants are often contaminated by organic pollutants such as polycyclic aromatic hydrocarbons (PAH). Their removal under methanogenic conditions was already reported, but the factors influencing this removal remain unclear. Here, we determined the influence of microbial communities on PAH removal under controlled physico-chemical conditions. Twelve mesophilic anaerobic digesters were inoculated with three microbial communities extracted from ecosystems with contrasting pollution histories: a PAH contaminated soil, a PCB contaminated sediment and a low contaminated anaerobic sludge. These anaerobic digesters were operated during 100 days in continuous mode. A sterilised activated sludge, spiked with 13 PAH at concentrations usually encountered in full-scale wastewater treatment plants, was used as substrate. The dry matter and volatile solid degradation, the biogas production rate and composition, the volatile fatty acids (VFA) production and the PAH removals were monitored. Bacterial and archaeal communities were compared in abundance (qPCR), in community structure (SSCP fingerprinting) and in dominant microbial species (454-pyrosequencing). The bioreactors inoculated with the community extracted from low contaminated anaerobic sludge showed the greater methane production. The PAH removals ranged from 10 % to 30 %, respectively, for high and low molecular weight PAH, whatever the inoculums tested, and were highly correlated with the dry matter and volatile solid removals. The microbial community structure and diversity differed with the inoculum source; this difference was maintained after the 100 days of digestion. However, the PAH removal was not correlated to these diverse structures and diversities. We hence obtained three functional stable consortia with two contrasted metabolic activities, and three different pictures of microbial diversity, but similar PAH and matter removals. These results confirm that PAH removal depends on the molecule type and on the solid matter removal. But, as PAH elimination is similar whether the solid substrate is degraded into VFA or into methane, it seems that the fermentative communities are responsible for their elimination.

Trace organic contaminants (TOCs) correspond to a broad range of molecules generated either directly or indirectly by human activity. Even though TOCs are found at low concentrations in the environment, they often accumulate by biomagnification and bioaccumulation into biological organisms and cause irreversible damages in biological systems through direct or indirect toxic effects such as endocrine disruption and tumour initiation. This manuscript presents the main findings of over fifteen years of research focusing on biological removal of various TOCs found in sewage sludge from urban treatment plants. A special focus of the research was made on microbial processes in complex anaerobic ecosystems. Four families of compounds mostly retrieved in urban plants were studied: the polycyclic aromatic hydrocarbons (PAHs), the polychlorobiphenyls (PCBs), the phthalic acid esters (PAEs), and the nonylphenol ethoxylates (NPEs). It was observed that the microbial capability for removing low amounts of TOCs required a long adaptation time and was often limited by the bioavailability of these compounds. In fact, the overall biodegradation resulted from the numerous interactions existing between the matrix (organic matter) and the microbial ecosystems according to the physico-chemical sorption properties of these compounds. Mechanistic aspects were also tackled in depth and specific models were developed for better understanding the network of interactions between TOCs, microorganisms, and organic matter. These findings could be extrapolated to other ecosystems such as soils and sediments. Finally, it was shown that microbial cometabolism was essential for TOC removal, and the concept of bioavailability was not only dependent on the nature, the level, and the sorption properties of TOCs but was also strongly dependent on the nature and the concentration of the sludge organic matter. Specific parameters were proposed for better evaluating the fate of TOCs in microbial anaerobic processes and technological solutions for efficient removal of these compounds were also proposed. removal of trace organic contaminants in sewage sludge: 15 years of experience. Pedosphere. 22(4): 508-517.