Abstract

A combination of culture-dependent and independent methods was used to study the co-existence of different sulfate-reducing bacteria (SRB) in an upflow anaerobic sludge bed reactor treating sulfate-rich wastewater. The wastewater was fed with ethanol as an external electron donor. Twenty six strains of SRB were randomly picked and isolated from the highest serial dilution that showed growth (i.e. 108). Repetitive enterobacterial palindromic polymerase chain reaction and whole cell protein profiling revealed a low genetic diversity, with only two genotypes among the 26 strains obtained in the pure culture. The low genetic diversity suggests the absence of micro-niches within the reactor, which might be due to a low spatial and temporal micro-heterogeneity. The total 16S rDNA sequencing of two representative strains L3 and L7 indicated a close relatedness to the genus Desulfovibrio. The two strains differed in as many as five physiological traits, which might allow them to occupy distinct niches and thus co-exist within the same habitat. Whole cell hybridisation with fluorescently labeled oligonucleotide probes was performed to characterise the SRB community in the reactor. The isolated strains Desulfovibrio L3 and Desulfovibrio L7 were the most dominant SRB, representing 30–35% and 25–35%, respectively, of the total SRB community. Desulfobulbus-like bacteria contributed for 20–25%, and the Desulfobacca acetoxidans-specific probe targeted approximately 15–20% of the total SRB. The whole cell hybridisation results thus revealed a consortium of four different species of SRB that can be enriched and maintained on a single energy source in a full-scale sulfidogenic reactor.

Highlights

  • Many industrial processes, such as metal smelting, flue gas scrubbing and mining generate sulfate-rich wastewater (Lens et al 1998)

  • We investigated the co-existence of physiologically similar hydrogenotrophic sulfate-reducing bacteria (SRB) in a full-scale sulfidogenic bioreactor treating sulfate-rich wastewater using a combination of cultivation and molecular techniques

  • Granular sludge was obtained from an upflow anaerobic sludge bed (UASB) reactor, treating sulfate-rich wastewater from a chemical plant located in Emmen, The Netherlands

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Summary

Introduction

Many industrial processes, such as metal smelting, flue gas scrubbing and mining generate sulfate-rich wastewater (Lens et al 1998). These wastewaters usually do not contain any organic carbon, and the addition of external electron donor is required for their treatment. Bioreactor processes have been developed for treating these sulfate and metal-rich wastewaters. This technology developed by the company Paques BV in The Netherlands (Pol et al 2001) makes use of the dissimilatory sulfate-reducing capacity of sulfate-reducing bacteria (SRB) to simultaneously remove sulfate and metals in the form of metal sulfides. The second step is the biological oxidation of the remaining sulfide to insoluble elemental sulfur, which is either recovered by sedimentation or a small portion

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