The effect of heavy metals on microbial community structure of a sulfidogenic consortium in anaerobic semi-continuous stirred tank reactors

Abstract

The effect of heavy metals on community structure of a heavy metal tolerant sulfidogenic consortium was evaluated by using a combination of denaturing gradient gel electrophoresis (DGGE) of 16S rRNA gene and dissimilatory sulfite reductase (dsrB) gene fragments, 16S rRNA gene cloning analysis and fluorescence in situ hybridization (FISH). For this purpose, four anaerobic semi-continuous stirred tank reactors (referred as R1-R4) were run in parallel for 12weeks at heavy metal loading rates of 1.5, 3, 4.5 and 7.5mgl(-1)d(-1) each of Cu(2+), Ni(2+), Zn(2+), and Cr(6+), respectively. The abundance ratio of Desulfovibrio vulgaris detected by FISH to total cell counts was consistent with the obtained results of cloning and DGGE. This indicated that D. vulgaris was dominant in all analyzed samples and played a key role in heavy metal removal in R1, R2, and R3. In contrast, after 4weeks of operation of R4, a distinct biomass loss was observed and no positive hybridized cells were detected by specific probes for the domain Bacteria, sulfate-reducing bacteria and D. vulgris. High removal efficiencies of heavy metals were achieved in R1, R2 and R3 after 12weeks, whereas the precipitation of heavy metals in R4 was significantly decreased after 4weeks and almost not observed after 6weeks of operation. In addition, the anaerobic bacteria, such as Pertrimonas sulfuriphila, Clostridium sp., Citrobacter amalonaticus, and Klebsiella sp., identified from DGGE bands and clone library were hypothesized as heavy metal resistant bacteria at a loading rate of 1.5mgl(-1)d(-1) of Cu(2+), Ni(2+), Zn(2+), and Cr(6+.)