Microbial analysis of in situ biofilm formation in drinking water distribution systems: implications for monitoring and control of drinking water quality.

Biofilm formation in drinking water distribution systems (DWDS) is influenced by the source water, the supply infrastructure and the operation of the system. A holistic approach was used to advance knowledge on the development of mixed species biofilms in situ, by using biofilm sampling devices installed in chlorinated networks. Key physico-chemical parameters and conventional microbial indicators for drinking water quality were analysed. Biofilm coverage on pipes was evaluated by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). The microbial community structure, bacteria and fungi, of water and biofilms was assessed using pyrosequencing. Conventional wisdom leads to an expectation for less microbial diversity in groundwater supplied systems. However, the analysis of bulk water showed higher microbial diversity in groundwater site samples compared with the surface water site. Conversely, higher diversity and richness were detected in biofilms from the surface water site. The average biofilm coverage was similar among sites. Disinfection residual and other key variables were similar between the two sites, other than nitrates, alkalinity and the hydraulic conditions which were extremely low at the groundwater site. Thus, the unexpected result of an exceptionally low diversity with few dominant genera (Pseudomonas and Basidiobolus) in groundwater biofilm samples, despite the more diverse community in the bulk water, is attributed to the low-flow hydraulic conditions. This finding evidences that the local environmental conditions are shaping biofilm formation, composition and amount, and hence managing these is critical for the best operation of DWDS to safeguard water quality

Selective enrichment and production of highly urease active bacteria by non-sterile (open) chemostat culture

In general, bioprocesses can be subdivided into naturally occurring processes, not requiring sterility (e.g., beer brewing, wine making, lactic acid fermentation, or biogas digestion) and other processes (e.g., the production of enzymes and antibiotics) that typically require a high level of sterility to avoid contaminant microbes overgrowing the production strain. The current paper describes the sustainable, non-sterile production of an industrial enzyme using activated sludge as inoculum. By using selective conditions (high pH, high ammonia concentration, and presence of urea) for the target bacterium, highly active ureolytic bacteria, physiologically resembling Sporosarcina pasteurii were reproducibly enriched and then continuously produced via chemostat operation of the bioreactor. When using a pH of 10 and about 0.2 M urea in a yeast extract-based medium, ureolytic bacteria developed under aerobic chemostat operation at hydraulic retention times of about 10 h with urease levels of about 60 μmol min-1 ml-1 culture. For cost minimization at an industrial scale the costly protein-rich yeast extract medium could be replaced by commercial milk powder or by lysed activated sludge. Glutamate, molasses, or glucose-based media did not result in the enrichment of ureolytic bacteria by the chemostat. The concentration of intracellular urease was sufficiently high such that the produced raw effluent from the reactor could be used directly for biocementation in the field

Micro-scale observations of the structure of aerobic microbial granules used for the treatment of nutrient-rich industrial wastewater

The structure and function of aerobic microbial granules from a lab-scale sequencing batch reactor treating nutrient-rich abattoir wastewater were investigated. These wastewater-fed granules were examined using a wide range of micro-scale techniques including light microscopy, scanning and transmission electron microscopy, fluorescent in situ hybridisation (FISH) combined with confocal laser scanning microscopy and oxygen and pH microsensors, in conjunction with a range of measurements in the bulk liquid phase. Interesting structural features were observed in these granules that have not been reported in synthetic-fed granules. The complex nature of abattoir wastewater was suggested to be responsible for accelerating the breaking process of large mature granules due to a rapid clogging of the granules pores and channels and for the very diverse microbial community observed displaying specific spatial distribution throughout the granules. More importantly, the dissolution at lower pH of mineral complexes associated to the granule matrix of extracellular polymeric substances might have caused the structural damages observed on the granules even though some pH buffer capacity was observed inside these granules. Ciliate protozoa were found to be very abundant on the surface of these wastewater-fed granules, which could potentially assist with reducing the high levels of suspended solids usually present in the aerobic granular sludge effluent. All these observations provide support to future studies on aerobic granular sludge treating real wastewater especially with regard to the granule structure and the mechanisms involved in their formation