Much attention is placed on organohalide-respiring bacteria (OHRB), such as Dehalococcoides, during the design and performance monitoring of chlorinated solvent bioremediation systems. However, many OHRB cannot function effectively without the support of a diverse group of other microbial community members (MCMs), who play key roles fermenting organic matter into more readily useable electron donors, producing corrinoids such as vitamin B12, or facilitating other important metabolic processes or biochemical reactions. While it is known that certain MCMs support dechlorination, a metric considering their contribution to bioremediation performance has yet to be proposed. Advances in molecular biology tools offer an opportunity to better understand the presence and activity of specific microbes, and their relation to bioremediation performance. In this paper, we test the hypothesis that a specific microbial consortium identified within 16S ribosomal ribonucleic acid (rRNA) gene next generation sequencing (NGS) data can be predictive of contaminant degradation rates. Field-based data from multiple contaminated sites indicate that increasing relative abundance of specific MCMs correlates with increasing first-order degradation rates. Based on these results, we present a framework for computing a simplified metric using NGS data, the Microbial Community Structure Index, to evaluate the adequacy of the microbial ecosystem during assessment of bioremediation performance.

Due to its adverse health and environmental impacts, groundwater contamination by toxic organic compounds such as chlorinated solvents is a global concern. The slow-release permanganate gel (SRP-G) is a mixture of potassium permanganate (KMnO4) and colloidal silica solution. The SRP-G is designed to radially spread after injection via wells, gelate in situ to form gel barriers containing permanganate (MnO4-), and slowly release MnO4- to treat plumes of chlorinated solvents in groundwater. This study aimed to characterize the effects of temperature on the dynamics of SRP-G in saturated porous media. In gelation batch tests, the viscosity of ambient-temperature (24 °C) SRP-G with 30 g/L-KMnO4 was 21 cP at 70 min, 134 cP at 176 min, and peaked at 946 cP to solidification at 229 min. The viscosity of low-temperature (4 °C) SRP-G with 30 g/L-KMnO4 was 71 cP at 273 min, 402 cP at 392 min, and peaked at 818 cP to solidification at 485 min. A similar pattern, e.g., increased gelation lag time with low-temperature SRP-G, was observed for SRP-Gs with 40 g/L, 50 g/L, and 60 g/L KMnO4. In flow-through tests using a glass column filled with saturated sands, injection rates, spreading rates, and release durations were 0.6 mL/min, 46 mm/min, and 33 h for KMnO4(aq), 0.2 mL/min, 2 mm/min, and 38 h for ambient-temperature SRP-G, and 0.4 mL/min, 16 mm/min, and 115 h for low-temperature SRP-G, respectively. These results indicated that the injectability, injection rate, and gelation lag time of SRP-G and the size, release rate, and release duration of MnO4- gel barriers can be increased at low temperatures. The low-temperature SRP-G scheme can be useful for treating large or dilute dissolved plumes of chlorinated solvents or other pollutants in groundwater.

Groundwater contamination by 1,2,3-trichloropropane (TCP) poses a unique challenge due to its human toxicity and recalcitrance to degradation. Previous work suggests that nitrogenous functional groups of pyrogenic carbonaceous matter (PCM), such as biochar, are important in accelerating contaminant dechlorination by sulfide. However, the reaction mechanism is unclear due, in part, to PCM\'s structural complexity. Herein, PCM-like polymers (PLPs) with controlled placement of nitrogenous functional groups [i.e., quaternary ammonium (QA), pyridine, and pyridinium cations (py+)] were employed as model systems to investigate PCM-enhanced TCP degradation by sulfide. Our results suggest that both PLP-QA and PLP-py+ were highly effective in facilitating TCP dechlorination by sulfide with half-lives of 16.91 ± 1.17 and 0.98 ± 0.15 days, respectively, and the reactivity increased with surface nitrogenous group density. A two-step process was proposed for TCP dechlorination, which is initiated by reductive ß-elimination, followed by nucleophilic substitution by surface-bound sulfur nucleophiles. The TCP degradation kinetics were not significantly affected by cocontaminants (i.e., 1,1,1-trichloroethane or trichloroethylene), but were slowed by natural organic matter. Our results show that PLPs containing certain nitrogen functional groups can facilitate the rapid and complete degradation of TCP by sulfide, suggesting that similarly functionalized PCM might form the basis for a novel process for the remediation of TCP-contaminated groundwater.

The complexity of the subsurface contaminated by chlorinated solvents such as trichloroethylene (TCE) makes it challenging to gain a complete understanding of contamination distribution and establish a conceptual site model (CSM). High-resolution vertical contaminant concentration profiling across both the unsaturated zone and the saturated aquifer is desirable for mapping the distribution of contamination. A Fick\'s law-based polydimethylsiloxane (PDMS) dialysis passive sampler was developed and evaluated on a field scale for its potential application. This study tests the passive sampler at two TCE contaminated sites, and the sampling results were compared with the results from different sampling methods based on the relative percent difference. The PDMS dialysis passive sampler obtained more representative soil gas concentrations in the unsaturated zone than a portable monitoring and sampling device, which caused soil gas flow disturbance by soil gas pumping during sample collection. In the saturated aquifer sampling, the results obtained by the PDMS dialysis passive sampler correlated well with those obtained by a commercial polyethylene passive diffusion bag, and exhibited higher sensitivity under low TCE concentration conditions. Furthermore, the PDMS dialysis passive samplers were densely deployed inside each monitoring well at multiple depths, at two sites, to achieve high-resolution monitoring across the unsaturated zone and saturated aquifer. Based on the PDMS dialysis sampler data, a more comprehensive three-dimensional CSM was systematically established.

Cocontamination by multiple chlorinated solvents is a prevalent issue in groundwater, presenting a formidable challenge for effective remediation. Despite the recognition of this issue, a comprehensive assessment of microbial detoxification processes involving chloroethenes and associated cocontaminants, along with the underpinning microbiome, remains absent. Moreover, strategies to mitigate the inhibitory effects of cocontaminants have not been reported. Here, we revealed that chloroform exhibited the most potent inhibitory effects, followed by 1,1,1-trichloroethane and 1,1,2-trichloroethane, on dechlorination of dichloroethenes (DCEs) in Dehalococcoides-containing consortia. The observed inhibition could be attributed to suppression of biosynthesis and enzymatic activity of reductive dehalogenases and growth of Dehalococcoides. Notably, cocontaminants more profoundly inhibited Dehalococcoides populations harboring the vcrA gene than those possessing the tceA gene, thereby explaining the accumulation of vinyl chloride under cocontaminant stress. Nonetheless, we successfully ameliorated cocontaminant inhibition by augmentation with Desulfitobacterium sp. strain PR owing to its ability to attenuate cocontaminants, resulting in concurrent detoxification of DCEs, trichloroethanes, and chloroform. Microbial community analyses demonstrated obvious alterations in taxonomic composition, structure, and assembly of the dechlorinating microbiome in the presence of cocontaminants, and introduction of strain PR reshaped the dechlorinating microbiome to be similar to its original state in the absence of cocontaminants. Altogether, these findings contribute to developing bioremediation technologies to clean up challenging sites polluted with multiple chlorinated solvents.

Biodegradation and biotransformation of contaminants in groundwater commonly occurs naturally. However, natural biodegradation rates can be slow leading to elongated contaminant plumes and prolonged risks that demand greater remedial intervention. Enhancement of the biodegradation of contaminants in groundwater can be induced by the addition of amendments to change the geochemical conditions to those that are more favorable for indigenous or added biota. Enhancing biodegradation requires collocation of the contaminant of concern with the \'right\' microbial communities under the \'right\' geochemical conditions, so that the microbiota thrive and bio-transform, degrade or lock up the contaminant of interest. This is most easily achievable at laboratory or bench scale where mixing is easily performed, and mass transfer limitations are minimized. However, inducing such changes at field scale in aquifers is non-trivial - amendments do not easily mix into groundwater because it is a laminar (non-turbulent) and low-energy flow environment. Bioaugmentation of cultured or genetically modified organisms have also been considered to add to groundwater to enhance contaminant degradation rates. Here we provide an overview of research studies over approximately 40 years that highlight the progression of understanding from natural biodegradation of plumes in groundwater to active bioremediation efforts that have been variably successful at field scale. Investigated contaminants providing insights include petroleum hydrocarbons, chlorinated and brominated hydrocarbons, ammonium, metals, munition compounds, atrazine and per- and polyfluorinated alkyl substances. The redox and electron acceptor/donor conditions that are inducive to biodegradation for a range of contaminants are highlighted. Biodegradation is challenged by the availability of electron donors/acceptors in the core of plumes and on plume fringes. Cases for bioaugmentation are identified. A long history of investigations provides examples of the importance of amendment delivery mechanisms, scale-up from laboratory to field, and field-scale demonstration of the effectiveness of groundwater bioremediation technologies. Advantages and disadvantages of remedial approaches are tabulated. The value and contributions of integrative modelling advances are identified. The literature review and example cases provide a deep understanding of what scale of bioremediation might be achievable for groundwater plumes. Limitations to bioremediation strategies outlined here will help direct future efforts. Addressing the sources of groundwater plumes as well as bioremediation of the plume itself will achieve more effective outcomes. Twelve \'lessons learnt\' are synthesized from the review.

To assess the efficiency of remediating dense non-aqueous phase liquids (DNAPLs), here heavy chlorinated solvents, through injection of xanthan solutions with or without surfactant (sodium dodecylbenzenesulfonate: SDBS), we conducted a comprehensive investigation involving rheological measurements, column (1D) and two-dimensional (2D) sandbox experiments, as well as numerical simulations on two-layers sand packs. Sand packs with grain sizes of 0.2-0.3 mm and 0.4-0.6 mm, chosen to represent the low and high permeable soil layers respectively, were selected to be representative of real polluted field. The rheological analysis of xanthan solutions showed that the addition of SDBS to the solution reduced its viscosity due to repulsive electrostatic forces and hydrophobic interactions between the molecules while preserving its shear-thinning behavior. Results of two-phase flow experiments depicted that adding SDBS to the polymer solution led to a reduced differential pressure along the soil and improvements of the DNAPL recovery factor of approximately 0.15 and 0.07 in 1D homogeneous and 2D layered systems, respectively. 2D experiments revealed that the displacement of DNAPL in multilayer zones was affected by permeability difference and density contrast in a heterogeneous soil. Simulation of multiphase flow in a multilayered system was performed by incorporating non-Newtonian properties and coupling the continuity equation with generalized Darcy\'s law. The results of modeling and experiments are very consistent. Numerical simulations showed that for an unconfined soil, the recovery of DNAPL by injection of xanthan solution can be reduced for more than 50%. In a large 2D experimental system, the combination of injecting xanthan with blocking the contaminated zone led to a promising remediation of DNAPL-contaminated layered zones, with a recovery of 0.87.

Perfluoroalkyl acids (PFAAs) have been shown to inhibit biodegradation (i.e., organohalide respiration) of chlorinated ethenes. The potential negative impacts of PFAAs on microbial species performing organohalide respiration, particularly Dehalococcoides mccartyi (Dhc), and the efficacy of in situ bioremediation are a critical concern for comingled PFAA-chlorinated ethene plumes. Batch reactor (no soil) and microcosm (with soil) experiments, containing a PFAA mixture and bioaugmented with KB-1, were completed to assess the impact of PFAAs on chlorinated ethene organohalide respiration. In batch reactors, PFAAs delayed complete biodegradation of cis-1,2-dichloroethene (cis-DCE) to ethene. Maximum substrate utilization rates (a metric for quantifying biodegradation rates) were fit to batch reactor experiments using a numerical model that accounted for chlorinated ethene losses to septa. Fitted values for cis-DCE and vinyl chloride biodegradation were significantly lower (p < 0.05) in batch reactors containing ≥50 mg/L PFAAs. Examination of reductive dehalogenase genes implicated in ethene formation revealed a PFAA-associated change in the Dhc community from cells harboring the vcrA gene to those harboring the bvcA gene. Organohalide respiration of chlorinated ethenes was not impaired in microcosm experiments with PFAA concentrations of 38.7 mg/L and less, suggesting that a microbial community containing multiple strains of Dhc is unlikely to be inhibited by PFAAs at lower, environmentally relevant concentrations.

In this work, chloride ions were used as conservative tracers and supplemented with conservative amounts of chloroethenes (PCE, TCE, Cis-DCE, 1,1-DCE), chloroethanes (1,1,1-TCA, 1,1-DCA), and the carbon isotope ratios of certain compounds, the most representative on the sites studied, which is a novelty compared to the optimization methods developed in the scientific literature so far. A location of the potential missing sources is then proposed in view of the balances of the calculated mixing fractions. A test of the influence of measurement errors on the results shows that the uncertainties in the calculation of the mixture fractions are less than 11%, indicating that the source identification method developed is a robust tool for identifying sources of chlorinated solvents in groundwater.

Aromatic oligoamide sequences programmed to fold into stable helical conformations were designed to display a linear array of hydrogen-bond donors and acceptors at their surface. Sequences were prepared by solid-phase synthesis. Solution 1 H NMR spectroscopic studies and solid-state crystallographic structures demonstrated the formation of stable hydrogen-bond-mediated dimeric helix bundles that could be either heterochiral (with a P and an M helix) or homochiral (with two P or two M helices). Formation of the hetero- or homochiral dimers could be driven quantitatively using different chlorinated solvents-exemplifying a remarkable case of either social or narcissistic chiral self-sorting or upon imposing absolute handedness to the helices to forbid PM species.