Mixed-species biofilms cultured from an oil sand tailings pond can biomineralize metals.

Here, we used an in vitro biofilm approach to study metal resistance and/or tolerance of mixed-species biofilms grown from an oil sand tailings pond in northern Alberta, Canada. Metals can be inhibitory to microbial hydrocarbon degradation. If microorganisms are exposed to metal concentrations above their resistance levels, metabolic activities and hydrocarbon degradation can be slowed significantly, if not inhibited completely. For this reason, bioremediation strategies may be most effective if metal-resistant microorganisms are used. Viability was measured after exposure to a range of concentrations of ions of Cu, Ag, Pb, Ni, Zn, V, Cr, and Sr. Mixed-species biofilms were found to be extremely metal resistant; up to 20 mg/L of Pb, 16 mg/L of Zn, 1,000 mg/L of Sr, and 3.2 mg/L of Ni. Metal mineralization was observed by visualization with scanning electron microscopy with metal crystals of Cu, Ag, Pb, and Sr exuding from the biofilms. Following metal exposure, the mixed-species biofilms were analyzed by molecular methods and were found to maintain high levels of species complexity. A single species isolated from the community (Rhodococcus erythropolis) was used as a comparison against the mixed-community biofilm and was seen to be much less tolerant to metal stress than the community and did not biomineralize the metals.

Evaluation of microbial biofilm communities from an Alberta oil sands tailings pond.

Bitumen extraction from the oil sands of Alberta has resulted in millions of cubic meters of waste stored on-site in tailings ponds. Unique microbial ecology is expected in these ponds, which may be key to their bioremediation potential. We considered that direct culturing of microbes from a tailings sample as biofilms could lead to the recovery of microbial communities that provide good representation of the ecology of the tailings. Culturing of mixed species biofilms in vitro using the Calgary Biofilm Device (CBD) under aerobic, microaerobic, and anaerobic growth conditions was successful both with and without the addition of various growth nutrients. Denaturant gradient gel electrophoresis and 16S rRNA gene pyrotag sequencing revealed that unique mixed biofilm communities were recovered under each incubation condition, with the dominant species belonging to Pseudomonas, Thauera, Hydrogenophaga, Rhodoferax, and Acidovorax. This work used an approach that allowed organisms to grow as a biofilm directly from a sample collected of their environment, and the biofilms cultivated in vitro were representative of the endogenous environmental community. For the first time, representative environmental mixed species biofilms have been isolated and grown under laboratory conditions from an oil sands tailings pond environment and a description of their composition is provided.

Efficient activation of Valpha14 invariant NKT cells by foreign lipid antigen is associated with concurrent dendritic cell-specific self recognition.

A burst release of cytokines by Valpha14 invariant NKT (iNKT) cells upon their TCR engagement critically regulates innate and adaptive immune responses. However, it remains unclear in vivo why iNKT cells respond efficiently to microbial or intracellular lipid Ags that are at low levels or that possess suboptimal antigenicity. We found that dendritic cells (DCs) potentiated iNKT cells to respond to a minimal amount of ligand alpha-galactosylceramide (alphaGalCer) through CD1d-dependent autoreactive responses that require endosomal processing and CD1d trafficking. The ability of potentiation of NKT cells was DC specific and did not depend on costimulatory signals and IL-12 production by DCs. However, DCs that failed to synthesize a major endogenous lipid Ag isoglobotrihexosylceramide were unable to potentiate NKT cells for efficient activation. Further analysis showed that differences in the level and pattern of endogenous lipid Ag presentation differentiate DCs and B cells for effective potentiation and subsequent activation of iNKT cells in the presence of an exogenous Ag. Thus, CD1d-dependent potentiation by DCs may be crucial for iNKT cell-mediated immunity against infectious agents.