The toxicity of N-nitrosamines, their presence in drinking and environmental water supplies, and poorly understood recalcitrance collectively necessitate a better understanding of their potential for bioattenuation. Here, we show that the bacterial strain Rhodococcus jostii RHA1 can biotransform N-nitrosodiethylamine (NDEA), N-nitrosodi-n-propylamine (NDPA), N-nitrosopyrrolidine (NPYR), and possibly N-nitrosomorpholine (NMOR) in addition to N-nitrosodimethylamine (NDMA). Growth of cells on propane as the sole carbon source greatly enhanced degradation rates when contrasted with cells grown on complex organics. Propane-induced rates in order of fastest to slowest were NDMA > NDEA > NDPA > NPYR > NMOR at concentrations <2000 μg/L. Removal rates for linear functional groups scaled inversely with mass and cyclic nitrosamines were more recalcitrant than linear nitrosamines. Controls demonstrated significant NDEA and NDPA losses independent of biomass, suggesting abiotic processes may play a role in attenuation of these two compounds under experimental conditions tested here. In contrast to NDMA, a transition from first to zero order kinetics was not observed for the other nitrosamines included in this study over a concentration range of 20-2000 μg/L. A genetic knockout for the propane monooxygenase enzyme (PrMO) confirmed the role of this enzyme in the biotransformation of NDEA and NPYR. This study furthers our understanding of environmental nitrosamine attenuation by revealing an enzymatic mechanism for the biotransformation of multiple nitrosamines, their relative recalcitrance to transformation, and potential for abiotic loss.

Water treatment combining advanced oxidative processes with subsequent exposure to biological activated carbon (BAC) holds promise for the attenuation of recalcitrant pollutants. Here we contrast oxidation and subsequent biofiltration of treated wastewater effluent employing either ozone or UV/HO followed by BAC during pilot-scale implementation. Both treatment trains largely met target water quality goals by facilitating the removal of a suite of trace organics and bulk water parameters. N-nitrosodimethylamine (NDMA) formation was observed in ozone fed BAC columns during biofiltration and to a lesser extent in UV/HO fed columns and was most pronounced at 20 min of empty bed contact time (EBCT) when compared to shorter EBCTs evaluated. While microbial populations were highly similar in the upper reaches, deeper samples revealed a divergence within and between BAC filtration systems where EBCT was identified to be a significant environmental predictor for shifts in microbial populations. The abundance of Nitrospira in the top samples of both columns provides an explanation for the oxidation of nitrite and corresponding increases in nitrate concentrations during BAC transit and support interplay between nitrogen cycling with nitrosamine formation. The results of this study demonstrate that pretreatments using ozone versus UV/HO impart modest differences to the overall BAC microbial population structural and functional attributes, and further highlight the need to evaluate NDMA formation prior to full-scale implementation of BAC in potable reuse applications.

Biologically activated carbon (BAC) can be used in both municipal water and hazardous waste remediation applications to enhance contaminant attenuation in water; however, questions remain about how selective pressures can be applied to increase the capabilities of microbial communities to attenuate recalcitrant contaminants. Here we utilized flow-through laboratory columns seeded with municipally derived BAC and exposed to water from a local drinking water facility to query how propane biostimulation impacts resident microorganisms. Ecological analyses using high throughput phylogenetic sequencing revealed that while propane did not increase the total number of microbiological species, it did select for bacterial communities that were distinct from those without propane. Temporal extractions demonstrated that microbial succession was rapid and established in approximately 2 months. A higher density of propane monooxygenase genes and bacterial clades including the Pelosinus and Dechloromonas genera suggest an enhanced potential for the degradation of persistent water pollutants in propane-stimulated systems. However, the ecological selective pressure was exhausted in less than 15 cm of transit in this flow-through scenario (25 hour retention) indicating a pronounced zonation that could limit the size of a biostimulated zone and require physical mixing, hydraulic manipulation, or other strategies to increase the spatial impact of biostimulation in flow-through scenarios.