Acidithiobacillus ferrooxidans is a chemolithoautotrophic, mesophilic Gram-negative bacterium able to oxidize ferrous iron, sulfur, and metal sulfides. It forms monolayer biofilms where extracellular polymeric substances are essential for cell attachment and metal sulfide leaching. High-throughput proteomics has been applied to study the early process of biofilm formation on pyrite by At. ferrooxidans ATCC 23270. After 24 h contact with the mineral, planktonic and sessile (biofilm) cell subpopulations were separated and proteins extracted. In total, 1319 proteins were detected in both samples. Sixty-two of these were found to be increased in biofilms. Additionally, 25 proteins were found to be decreased in the biofilm cell subpopulation. Three transcriptional factors were found to be increased or decreased among both cell subpopulations, suggesting their potential involvement in the regulation of these processes. Although no significant differences were observed for the known proteins related to ferrous iron and sulfur oxidation pathways among both cell subpopulations, the results presented here show that the early steps of At. ferrooxidans biofilm formation consist of a set of metabolic adaptations following cell attachment to the mineral surface. Functions such as extracellular polymeric substances biosynthesis seem to be pivotal. This first high-throughput proteomic study may also contribute to the annotation of several unknown At. ferrooxidans proteins found.

Heavy metals such as zinc cannot be degraded by microorganisms and form long contaminant plumes in groundwater. Conventional methods for remediating heavy metal-contaminated sites are for example excavation and pump-and-treat, which is expensive and requires very long operation times. This induced interest in new technologies such as in situ adsorption barriers for immobilization of heavy metal contamination. In this study, we present steps and criteria from laboratory tests to field studies, which are necessary for a successful implementation of an in situ adsorption barrier for immobilizing zinc. Groundwater and sediment samples from a contaminated site were brought to the lab, where the adsorption of zinc to Goethite nanoparticles was studied in batch and in flow-through systems mimicking field conditions. The Goethite nanoparticles revealed an in situ adsorption capacity of approximately 23 mg Zn per g Goethite. Transport experiments in sediment columns indicated an expected radius of influence of at least 2.8 m for the injection of Goethite nanoparticles. These findings were validated in a pilot-scale field study, where an in situ adsorption barrier of ca. 11 m × 6 m × 4 m was implemented in a zinc-contaminated aquifer. The injected nanoparticles were irreversibly deposited at the desired location within <24 h, and were not dislocated with the groundwater flow. Despite a constantly increasing inflow of zinc to the barrier and the short contact time between Goethite and zinc in the barrier, the dissolved zinc was effectively immobilized for ca. 90 days. Then, the zinc concentrations increased slowly downstream of the barrier, but the barrier still retained most of the zinc from the inflowing groundwater. The study demonstrated the applicability of Goethite nanoparticles to immobilize heavy metals in situ and highlights the criteria for upscaling laboratory-based determinants to field-scale.

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In this study, initial attachment to and biofilm formation of Sulfobacillusthermosulfidooxidans DSM 9293T on pyrite in the presence of Leptospirillumferriphilum DSM 14647T were investigated. Interactions of S.thermosulfidooxidansT and L.ferriphilumT were studied by means of monitoring attachment behavior and biofilm formation on pyrite. Our preliminary results showed that 1): Pre-established biofilms of L.ferriphilumT had effects on attachment of S. thermosulfidooxidansT to pyrite; 2): physical contact between cells of L. ferriphilumT and S. thermosulfidooxidansT on pyrite were visible 3): Pyrite leaching by cells of S. thermosulfidooxidansT was inhibited by the presence of inactive cells of L.ferriphilumT.

Arsenic contamination of groundwater occurs due to both geogenic and anthropogenic processes. Conventional arsenic remediation techniques require extraction of groundwater into pump-and-treat systems, which are expensive and require long operational times. Hence, there is a need for cost-effective remediation. In this study, we assessed and validated the in situ remediation of arsenic contamination in groundwater resources using permeable reactive barriers (PRBs) made of injectable, colloidal iron oxide nanoparticles in the laboratory and in field-scale pilot tests. Sand-packed, flow-through column studies were used in order to assess the sorption behavior of the iron oxide nanoparticles using field materials (sand, groundwater) in the laboratory. The breakthrough curves were analyzed using a reactive transport model considering linear and nonlinear adsorption isotherms and were fitted best with a chemical nonequilibrium consideration. The results were used to design a pilot-scale field test. The injected 28 m3 of nanoparticles (ca. 280 kg dry weight of iron oxide) were successfully delivered to the aquifer via an injection well. No mobile iron was detected downstream, confirming that a stable in situ barrier was formed that did not move with the groundwater flow. Arsenic concentrations in groundwater were reduced to the aimed 50% of the background value, despite the relatively short contact time between arsenic and the iron oxide in the barrier, due to the high flow velocity of 1.21 m/day. We compared the results of the laboratory and field tests and concluded that the single-parameter models based on retardation factor and/or adsorption capacity fail to predict the longevity of the barrier and the evolution of arsenic breakthrough with time, most likely because they do not consider the chemical nonequilibrium effects. Therefore, we propose that upscaling the laboratory findings to field design must be carried out with care and be coupled with detailed reactive transport models.