Search

The performance of on-site wastewater treatment systems (OWTSs) can be improved by altering digester design and by manipulating environmental variables that affect microbial community composition. Community composition can potentially be assessed using high-throughput DNA sequencing, but the two most common methods of community DNA sequencing (16S and shotgun sequencing) generally yield different taxonomic identification profiles and can perform differently according to the sampled environment. To evaluate the use of these two approaches in monitoring OWTS operation, we conducted a comparative parallel analysis using both 16S rDNA and shotgun sequencing in a controlled field study. Results indicate that when examining microorganisms above 0.1% relative abundance, 16S and shotgun sequencing produced similar results in terms of individual sample species richness and between-sample community similarity. However, shotgun sequencing provided comparatively higher taxonomic richness for the bacterial communities with lower abundance in the OWTSs. In addition, 16S sequencing resolved only 48 out of 188 bacterial communities identified by shotgun sequencing (using a 0.1% abundance cutoff). Three key bacterial genera (Desulfomicrobium, Simplicispira, and Phenylobacterium) in anaerobic digestion processes were differentially abundant for both sequencing methods. These data indicate that both sequencing methods provide similar overall profiles for bacterial communities in anaerobic digestor systems. However, shotgun sequencing provides significantly (p-value < 0.01) higher taxonomic richness overall. Thus, shotgun sequencing provides a more robust taxonomic and functional profile that can be used for the optimization of anaerobic digestor systems.

Nonaqueous phase liquids (NAPL, e.g., hydrocarbons and chlorinated compounds) are common groundwater pollutants. Electrokinetic remediation of NAPLs uses electric fields to draw them toward electrodes and remove them from groundwater. The treatment requires NAPL mobility. Emulsification increases mobility, but at a risk for downstream receptors. We propose using alkaline aqueous solutions of zein and graphene nanoparticles (GNP) to form conductive materials, which could also act as barriers to control NAPL migration. Alkaline zein-GNP solutions can be injected in the polluted soil and solidified by neutralizing the pH (e.g., with glacial acetic acid, GAA). Shear rheology experiments showed that zein-GNP composites were cohesive, and voltammetry showed that GNP increased electrical conductivity of zein-based materials by 3.5 times. Gas chromatography-mass spectroscopy (GC-MS) demonstrated that the electrokinetic treatment of model sandy aquifers yielded >60% and ∼47% removal of emulsified toluene in freshwater and in salt solutions, respectively (with 30 min treatment using a 10 V differential voltage between a zein-GNP and an aluminum electrode. NaCl was used as model salt contaminant. The conductivity of surfactant solutions was lower in saline water than in freshwater, explaining differences in toluene removal. Toluene-water emulsions were stabilized using the natural surfactants lecithin and saponin. These surfactants acted synergistically in stabilizing emulsions in either freshwater or salt solutions. Lecithin and saponin likely interacted at toluene-water interfaces, as indicated by the morphology, interfacial tension and compressional rigidity of toluene-water interfaces with both components (relative to interfaces of either lecithin or saponin alone). The compressional behavior of interfacial films was well-described by the Marczak model. Electrokinetic treatment of saturated model sandy aquifers also decreased the turbidity of emulsions of water and either tricholoroethylene (TCE, by ∼41%) or diesel (by ∼75%), in the presence of a bacterial biosurfactant. This decrease was used as semiquantitative indicator of NAPL removal from water.

Environmental water monitoring is an important responsibility of municipal governments. In this study, we partnered with several municipalities in an extensive sampling program to investigate the effects of spatiotemporal and environmental factors on bacterial diversity in a complex watershed ecosystem containing specific environments including creeks, a river, canals, stormwater outfalls and freshwater lakes of the Niagara Peninsula. Samples were collected using standard municipal protocols and bacterial DNA extracted from these samples was sequenced using high-throughput DNA sequencing targeting the V3–V4 regions of the 16S rRNA gene. Average taxonomic richness and alpha diversity differed significantly between samples collected from lakes and creeks (P

Residents in rural communities across Canada collect potable water from aquifers. Fecal contaminants from sewage and agricultural runoffs can penetrate aquifers, posing a public health risk. Standard methods for detecting fecal contamination test for fecal indicator bacteria (FIB), but the presence of these do not identify sources of contamination. In contrast, DNA-based diagnostic tools can achieve this important objective. We employed quantitative polymerase chain reaction (qPCR) and high-throughput DNA sequencing to trace fecal contamination sources in Wainfleet, a rural Ontario township that has been under the longest active boil water advisory in Canada due to FIB contamination in groundwater wells. Using traditional methods, we identified FIBs indicating persistent fecal pollution in well waters. We used 16S rRNA sequencing to profile groundwater microbial communities and identified Campylobacteraceae as a fecal contamination DNA marker in septic tank effluents (STEs). We also identified Turicibacter and Gallicola as a potential cow and chicken fecal contamination marker, respectively. Using human specific Bacteroidales markers, we identified leaking septic tanks as the likely primary fecal contamination source in some of Wainfleet's groundwater. Overall, the results support the use of sequencing-based methods to augment traditional water quality testing methods and help end-users assess fecal contamination levels and identify point and non-point pollution sources. This article has been made Open Access thanks to the generous support of a global network of libraries as part of the Knowledge Unlatched Select initiative.

This study proposes injectable polymeric filters to prevent diesel migration during its bioremediation. Pseudomonas aeruginosa (Gram negative bacterium) was isolated from forest soil and used to biodegrade diesel. P. aeruginosa produced biosurfactants that emulsified diesel in water while degrading it, by decreasing the interfacial tension and forming skins around diesel droplets. In lab experiments, emulsified diesel droplets could migrate through model soil columns (sand and limestone) saturated with water (to mimic aquifers). These results indicate that clean aquifers downstream of diesel spills are at risk of becoming contaminated during diesel bioremediation by P. aeruginosa. Injectable filters were developed to mitigate this risk. Injectable filters were obtained by injecting into model soils aqueous solutions of quaternized hydroxyethylcellulose (HEC+, with a positive electrostatic charge). HEC + adsorbed onto model soils, yielding positively charged surfaces. Diesel droplets stabilized by P. aeruginosa had a negative electrostatic charge (as demonstrated through zeta potential measurements) and adhered onto HEC + coated soil, likely due to attractive electrostatic forces. As a result, diesel droplet mobility through soil columns decreased. Our proof of concept indicates that injectable HEC + filters can protect clean regions of aquifers during the bioremediation of diesel spills by P. aeruginosa.

Methylenetetrahydrofolate models (green substructure) reduce organohalides to the respective hydrocarbons under biomimetic conditions and mimic the activity of dehalohydrogenases., Halocarbons R–X are reduced to hydrocarbons R–H by folate model compounds under biomimetic conditions. The reactions correspond to a halide–hydride exchange with the methylenetetrahydrofolate (MTHF) models acting as hydride donors. The MTHF models are also functional equivalents of dehalohydrogenases but, unlike these enzymes, do not require a metal cofactor. The reactions suggest that halocarbons have the potential to act as endocrinological disruptors of biochemical pathways involving MTHF. As a case in point, we observe the rapid reaction of the MTHF models with the inhalation anaesthetic halothane. The ready synthetic accessibility of the MTHF models as well as their dehalogenation activity in the presence of air and moisture allow for the remediation of toxic, halogenated hydrocarbons.

Pseudomonas sp. bacteria were incorporated into a double emulsion system (water in oil in water, W/O/W), which can be injected in the subsurface to trap and biotreat naphthalene. The outermost phase of the W/O/W emulsion was an aqueous solution of chitosan (1.65 wt.%). Naphthalene could be dispersed in the aqueous chitosan phase of W/O/W emulsions. Two separate primary water in oil (W/O) emulsions were emulsified in chitosan, using Tween 20 as emulsifier. W/O emulsions were prepared with canola oil (containing ethylcellulose and calcium stearate as emulsifiers), and aqueous solutions of either Pseudomonas sp. in minimal broth or sodium citrate (crosslinker). The formation of W/O/W double emulsions was verified with optical and confocal microscopy. Without addition of triggers able to induce gelation on demand, W/O/W emulsions were stable and mainly viscous under shear. The shear elastic modulus (G’ = 1.90 ± 0.39 Pa) and the shear viscous modulus (G” = 5.76 ± 0.06 Pa) were low, to facilitate pumping in the subsurface. With addition of the surfactant cocamidopropyl betaine, W/O/W emulsions were destabilized on demand, thereby releasing citrate (crosslinker) and Pseudomonas sp. in the chitosan phase, where naphthalene was dispersed. Mixing chitosan with citrate led to its gelation, trapping naphthalene. Mixing naphthalene with Pseudomonas sp. led to its bioremediation. Gelation was demonstrated by the increase in the viscoelastic moduli, with G’ = 111.02 ± 1.67 Pa > G” = 8.66 ± 0.10 Pa. With naphthalene, Pseudomonas sp. survival in gelled chitosan was confirmed by plating bacteria sampled from the gel. Without naphthalene, Pseudomonas sp. did not survive in chitosan.