Psychrophilic methanogenic community development during long-term cultivation of anaerobic granular biofilms

Granular biomass was temporally sampled from a cold (4-15 degrees C) anaerobic bioreactor, which was inoculated with mesophilic biomass and used to treat industrial wastewater in a long-term (3.4 year) study. Data from 16S rRNA gene clone libraries, quantitative PCR and terminal restriction fragment length polymorphism analyses indicated that microbial community structure was dynamic, with shifts in the archaeal and bacterial communities' structures observed following start-up and during temperature decreases from 15 to 9.5 degrees C (phase 1). Specifically, the relative abundance of architecturally important Methanosaeta-like (acetoclastic) methanogens decreased, which was concomitant with granule disintegration and the development of a putatively psychrophilic hydrogenotrophic methanogenic community. Genetic fingerprinting suggested the development of a psychroactive methanogenic community between 4 and 10 degrees C (phase 2), which was dominated by acetogenic bacteria and Methanocorpusculum-like (hydrogenotrophic) methanogens. High levels of Methanosaeta-like acetoclastic methanogens and granular biofilm integrity were maintained during phase 2. Overall, decreasing temperature resulted in distinctly altered microbial community structure during phase 1, and the development of a less dynamic psychroactive methanogenic consortium during phase 2. Moreover, psychrophilic H(2)-oxidizing methanogens emerged as important members of the psychroactive consortia after >1200 days of low-temperature cultivation. The data suggest that prolonged psychrophilic cultivation of mesophilic biomass can establish a well-functioning psychroactive methanogenic consortium, thus highlighting the potential of low-temperature anaerobic digestion technology.