Indigenous microbial communities in Albertan sediments are capable of anaerobic benzene biodegradation under methanogenic, sulfate-reducing, nitrate-reducing, and iron-reducing redox conditions.

Alberta is a major center for oil and gas production, and correspondingly harbors hundreds of unresolved contamination sites by environmental hazards such as benzene (C6 H6 ). Due to its cost-effectiveness, bioremediation has become a promising strategy for C6 H6 removal. Contamination sites typically take on an anaerobic context, which complicates the energetics of contamination sites and is a subject that is scarcely broached in studies of Albertan sediments. This study examines the innate potential for indigenous microbial communities in Albertan sediments to remove C6 H6 in a multitude of reduced conditions. Community profiles of these sediments were analyzed by 16S rRNA gene amplicon sequencing, and removal rates and reaction stoichiometries were observed by gas chromatography and ion chromatography. Organisms belonging to known primary degrader taxa were identified, including Geobacter (iron-reducing), and Peptococcaceae (nitrate-reducing). Furthermore, benzene removal patterns of the cultures were similar to those observed in previously reported microcosms, with lag times between 70 and 168 days and removal rates between 3.27 and 12.70 µM/day. Such information could support a more comprehensive survey of Albertan sediment consortia, which may eventually be utilized in informing future remediation efforts in the province. PRACTITIONER POINTS: ●Clay and sand sediments originating from Northern Alberta could remove benzene under methanogenic, sulfate-reducing, iron-reducing, and nitrate-reducing conditions. ●Degradation profiles were broadly comparable to those of reported cultures from other geographical locales. ●Key degrader taxa observed included Geobacter (Fe3+ -reducing) and Peptococcaceae ( NO 3 - -reducing). ●Knowledge gained can be the start of a more extensive survey of Albertan sediments. Eventually, this collection of information can be used to generate robust C6 H6 -degrading cultures that can be implemented for bioaugmentation and be implemented in informing remediation strategies in soil and water matrices for priority contamination cases such as leaking underground storage tanks and orphan wells.

Potential syntrophic associations in anaerobic naphthenic acids biodegrading consortia inferred with microbial interactome networks.

Naphthenic acids (NAs) can be syntrophically metabolized by indigenous microbial communities in pristine sediments beneath oil sands tailings ponds. Syntrophy is an essential determinant of the microbial interactome, however, the interactome network in anaerobic NAs-degrading consortia has not been previously addressed due to complexity and resistance of NAs. To evaluate the impact of electron acceptors on topology of interactome networks, we inferred two microbial interactome networks for anaerobic NAs-degrading consortia under nitrate- and sulfate-reducing conditions. The complexity of the network was higher under sulfate-reducing conditions than nitrate-reducing conditions. Differences in the taxonomic composition between the two modules implies that different potential syntrophic interactions exist in each network. We inferred the presence of the same syntrophic microorganisms, from genera Bellilinea, Longilinea, and Litorilinea, initiating the metabolism in both networks, but within each network, we predicted unique syntrophic associations that have not been reported. Electron acceptor has a large effect on the interactome networks for anaerobic NAs-degrading consortia, offers insight into an unrecognized dimension of these consortia. These results provide a novel approach for exploring potential syntrophic relationships in biodegrading processes to help cost-effectively remove NAs in oil sands tailings ponds.

Naphthenic acid anaerobic biodegrading consortia enriched from pristine sediments underlying oil sands tailings ponds.

Seepage from oil sands tailings ponds (OSTP), which contain toxic naphthenic acids (NAs), can infiltrate into groundwater. Clay sediment layer beneath is a critical barrier for reducing the infiltration of NAs into the sand sediment layer, where groundwater channels reside. Biodegradation has great potential as a strategy for NAs removal, but little is known about NAs biodegradability and potential functional microbes in these pristine sediments. This study investigated the potential for anaerobic biodegradation of NAs by microbial consortia enriched from clay and sand sediments underlying OSTP, amended with either acid extracted organics or Merichem NAs, under nitrate- and sulfate-reducing conditions. Degradation of NAs only be detected after DOC concentration reached to steady state after 163 days. Microbial community analysis shows that different electron acceptors, sediment types, and NAs sources associated with specific microbial taxa and can explain 14.8, 13.9 % and 5% of variation of microbial community structures, respectively. The DOC and methane were the most important geochemical properties for microbial community variations. This study approved the potential capability of indigenous microbial communities from the pristine sediments in NA degradation, demonstrating the barrier function of pristine clay sediments underlying OSTP in prohibiting organic contaminants from entering into groundwater.

Model naphthenic acids removal by microalgae and Base Mine Lake cap water microbial inoculum.

Naphthenic acids (NAs) originate from bitumen and are considered a major contributor to acute toxicity in oil sands process-affected water (OSPW) produced from bitumen extraction processes. To reclaim oil sands tailings and remediate OSPW, in-pit fluid fine tailings can be water-capped as end pit lakes (EPL). Addressing NAs present in OSPW, either through removal, dilution or degradation, is an objective for oil sands reclamation. EPLs can remediate NAs through degradation or dilution or both. To assess and understand degradation potential, Chlorella kessleri and Botryococcus braunii were tested for their tolerance to, and ability to biodegrade, three model NAs (cyclohexanecarboxylic acid, cyclohexaneacetic acid, and cyclohexanebutyric acid). Water sourced from the industry's first EPL, the Base Mine Lake (BML), was used alone as an inoculum or co-cultured with C. kessleri to biodegrade cyclohexanecarboxylic acid and cyclohexanebutyric acid. All cultures metabolized the model compounds via ß-oxidation. Biodegradation by the co-culture of C. kessleri and BML inoculum was most effective and rapid: the cyclohexaneacetic acid generated from cyclohexanebutyric acid could be further degraded by the co-culture, while the cyclohexaneacetic acid generated could not be consumed by pure algal cultures or BML inoculum alone. Adding C. kessleri greatly increased the diversity of the microbial community in the BML inoculum; many known hydrocarbon and NA degraders were identified from the 16S rRNA gene sequencing from this co-culture. This more diverse microbial community could have potential for EPL remediation.

Indigenous microorganisms residing in oil sands tailings biodegrade residual bitumen.

The purpose of this study was to determine the capacity of indigenous microbes in tailings to degrade bitumen aerobically, and if acetate biostimulation further improved degradation. Fluid fine tailings, from Base Mine Lake (BML), were used as microbial inocula, and bitumen in the tailings served as a potential carbon source during the experiment. The tailings were capped with 0.22 µm-filtered BML surface water with or without BML bitumen and acetate addition and incubated for 100 days at 20 °C. CO2 production and petroleum hydrocarbon reductions (50-70% for the biostimulation treatment) in the tailings were observed. DNA was extracted directly from the tailings, and increased bacterial density was observed by qPCR targeting the rpoB gene in the biostimulated group. 16 S rRNA sequencing was used to determine microbial composition profiles in each treatment group. The microbial communities indigenous to the tailings shifted after the bitumen was added. Acidovorax, Rhodoferax, Pseudomonas and Pseudoxanthomonas spp. significantly increased compared to the original microbial community and demonstrated tolerance to bitumen-based toxicity. The first three genera showed more potential for biostimulation treatment with acetate and may be important bitumen/hydrocarbon-degraders in an oil sands end pit lake environment.

Timed Regulation of 3BP2 Induction Is Critical for Sustaining CD8+ T Cell Expansion and Differentiation.

Successful anti-viral response requires the sustained activation and expansion of CD8+ T cells for periods that far exceed the time limit of physical T cell interaction with antigen-presenting cells (APCs). The expanding CD8+ T cell pool generates the effector and memory cell populations that provide viral clearance and long-term immunity, respectively. Here, we demonstrate that 3BP2 is recruited in cytoplasmic microclusters and nucleates a signaling complex that facilitates MHC:peptide-independent activation of signaling pathways downstream of the TCR. We show that induction of the adaptor molecule 3BP2 is a sensor of TCR signal strength and is critical for sustaining CD8+ T cell proliferation and regulating effector and memory differentiation.