Impacts of a herring gull colony on runoff water quality from an urban green roof.

Green infrastructure (GI) strategies, including green roofs, have become a common, decentralized, nature-based strategy for reducing urban runoff and restoring ecosystem services to the urban environment. In this study, we examined the water quality of incident rainfall and runoff from a green roof installed on top of the Jacob K. Javits Convention Center in New York City. Since the 2014 installation of this green roof, one of the largest in North America, a colony of nesting herring gulls grew to approximately 100 nesting pairs in 2018 and 150 nesting pairs in 2019. Water quality monitoring took place between September 2018 and October 2019. Except for phosphorus on some occasions, we found concentrations of nitrate, nitrite, chlorine, sulfate to be below federal drinking water standards. Levels of the fecal indicator bacteria (FIB), total coliform, E. coli, and Enterococcus, were consistently higher in runoff samples than rainwater, ranging from 150 to over 20,000 CFU/100 mL for E. coli and 100 to over 140,000 CFU/100 mL for total coliform. Quantitative polymerase chain reaction (qPCR) methods were used to search for potential opportunistic pathogens, including Legionella spp., Mycobacterium spp., Campylobacter spp., and Salmonella spp. Discovery of the presence of Catellicoccus marimammalium, a gull-associated marker in runoff water indicates that herring gulls are the likely source of contamination. Due to habitat loss, herring gulls, and other Larus gull species are increasingly nesting on urban roofs, both green (such as at the Javits Center) and conventional (such as on Rikers and Governors Islands). Habitat creation is one of the target ecosystem services desired from GI systems. Although the discharge from the green roof of the Javits Center is directed to the city's sewer system, this study demonstrates the need to treat runoff from green roofs with nesting gull populations if its intended use involves reuse or human contact.

Overcoming the inhibitory effects of urea to improve the kinetics of microbial-induced calcium carbonate precipitation (MICCP) by Lysinibacillus sphaericus strain MB284.

Among different microbial-induced calcium carbonate precipitation (MICCP) mechanisms utilized for biomineralization, ureolysis leads to the greatest yields of calcium carbonate. Unfortunately, it is reported that urea-induced growth inhibition can delay urea hydrolysis but it is not clear how this affects MICCP kinetics. This study investigated the impact of urea addition on the MICCP performance of Lysinibacillus sphaericus MB284 not previously grown on urea (thereafter named bio-agents), compared with those previously cultured in urea-rich media (20 g/L) (hereafter named bio-agents+ or bio-agents-plus). While it was discovered that initial urea concentrations exceeding 3 g/L temporarily hindered cell growth and MICCP reactions for bio-agents, employing bio-agents+ accelerated the initiation of bacterial growth by 33% and led to a 1.46-fold increase in the initial yield of calcium carbonate in media containing 20 g/L of urea. The improved tolerance of bio-agents+ to urea is attributed to the presence of pre-produced endogenous urease, which serves to reduce the initial urea concentration, alleviate growth inhibition, and expedite biomineralization. Notably, elevating the initial concentration of bio-agents+ from OD600 of 0.01 to 1, housing a higher content of endogenous urease, accelerated the initiation of MICCP reactions and boosted the ultimate yield of biomineralization by 2.6 times while the media was supplemented with 20 g/L of urea. These results elucidate the advantages of employing bio-agents+ with higher initial cell concentrations to successfully mitigate the temporary inhibitory effects of urea on biomineralization kinetics, offering a promising strategy for accelerating the production of calcium carbonate for applications like bio self-healing of concrete.

Impacts of divalent cations (Mg2+ and Ca2+) on PFAS bioaccumulation in freshwater macroinvertebrates representing different foraging modes.

Per- and polyfluoroalkyl substances (PFAS) have extensively contaminated freshwater aquatic ecosystems where they can be transported in water and partition to sediment and biota. In this paper, three freshwater benthic macroinvertebrates with different foraging modes were exposed to environmentally relevant concentrations of eight perfluoroalkyl carboxylates (PFCA), three perfluoroalkyl sulfonates (PFSA), and three fluorotelomer sulfonates (FTS) at varying divalent cation concentrations of magnesium (Mg2+) and calcium (Ca2+). Divalent cations can impact PFAS partitioning to solids, especially to sediments, at higher concentrations. Sediment dwelling worms (Lumbriculus variegatus), epibenthic grazing snails (Physella acuta), and sediment-dwelling filter-feeding bivalves (Elliptio complanata) were selected due to their unique foraging modes. Microcosms were composed of synthetic sediment, culture water, macroinvertebrates, and PFAS and consisted of a 28-day exposure period. L. variegatus had significantly higher PFAS bioaccumulation than P. acuta and E. complanata, likely due to higher levels of interactions with and ingestion of the contaminated sediment. "High Mg2+" (7.5 mM Mg2+) and "High Ca2+" (7.5 mM Ca2+) conditions generally had statistically higher bioaccumulation factors (BAF) than the "Reference Condition" (0.2 mM Ca2+ and 0.2 mM Mg2+) for PFAS with perfluorinated chain lengths greater than eight carbons. Long-chain PFAS dominated the PFAS profiles of the macroinvertebrates for all groups of compounds studied (PFCA, PFSA, and FTS). These results indicate that the study organism has the greatest impact on bioaccumulation, although divalent cation concentration had observable impacts between organisms depending on the environmental conditions. Elevated cation concentrations in the microcosms led to significantly greater bioaccumulation in the test organisms compared to the experimental reference conditions for long-chain PFAS.

Influence of microbial weathering on the partitioning of per- and polyfluoroalkyl substances (PFAS) in biosolids.

Per- and polyfluoroalkyl substances (PFAS) are a large group of man-made fluorinated organic chemicals that can accumulate in the environment. In water resource recovery facilities (WRRFs), some commonly detected PFAS tend to partition to and concentrate in biosolids where they can act as a source to ecological receptors and may leach to groundwater when land-applied. Although biosolids undergo some stabilization to reduce pathogens before land application, they still contain many microorganisms, contributing to the eventual decomposition of different components of the biosolids. This work demonstrates ways in which microbial weathering can influence biosolids decomposition, degrade PFAS, and impact PFAS partitioning in small-scale, controlled laboratory experiments. In the microbial weathering experiments, compound-specific PFAS biosolids-water partitioning coefficients (Kd) were demonstrated to decrease, on average, 0.4 logs over the course of the 91 day study, with the most rapid changes occurring during the first 10 days. Additionally, the highest rates of lipid, protein, and organic matter removal occurred during the same time. Among the evaluated independent variables, statistical analyses demonstrated that the most significant solids characteristics that impacted PFAS partitioning were organic matter, proteins, lipids, and molecular weight of organics. A multiple linear regression model was built to predict PFAS partitioning behavior in biosolids based on solid characteristics of the biosolids and PFAS characteristics with a R2 value of 0.7391 when plotting predicted and measured log Kd. The findings from this work reveal that microbial weathering can play a significant role in the eventual fate and transport of PFAS and their precursors from biosolids.

Bioaccumulation of per- and polyfluoroalkyl substances by freshwater benthic macroinvertebrates: Impact of species and sediment organic carbon content.

Per- and polyfluoroalkyl substances (PFAS) in aquatic environments have caused global concern due to their persistence, toxicity, and potential bioaccumulation of some compounds. As an important compartment of the aquatic ecosystem, sediment properties impact PFAS partitioning between aqueous and solid phases, but little is known about the influence of sediment organic carbon content on PFAS bioaccumulation in benthic organisms. In this study, three freshwater benthic macroinvertebrates - worms (Lumbriculus variegatus), mussels (Elliptio complanata) and snails (Physella acuta) - were exposed for 28 days to PFAS spiked synthetic sediment equilibrated with a synthetic surface water. Using microcosms, sediment organic carbon content - 2%, 5% and 8% - was manipulated to assess its impact on PFAS bioaccumulation. Worms were found to have substantially greater PFAS bioaccumulation compared to mussels and snails. The bioaccumulation factors (BAFs) and biota sediment accumulation factors (BSAFs) in worms were both one to two magnitudes higher than in mussels and snails, likely due to different habitat-specific uptake pathways and elimination capacities among species. In these experiments, increasing sediment organic carbon content decreased the bioaccumulation of PFAS to benthic macroinvertebrates. In worms, sediment organic carbon content was hypothesized to impact PFAS bioaccumulation by affecting PFAS partitioning and sediment ingestion rate. Notably, the BSAF values of 8:2 fluorotelomer sulfonic acid (FTS) were the largest among 14 PFAS for all species, suggesting that the benthic macroinvertebrates probably have different metabolic mechanisms for fluorotelomer sulfonic acids compared to fish evaluated in published literature. Understanding the impact of species and sediment organic carbon on PFAS bioaccumulation is key to developing environmental quality guidelines and evaluating potential ecological risks to higher trophic level species.

Plasma-generated reactive water mist for disinfection of N95 respirators laden with MS2 and T4 bacteriophage viruses.

Due to the shortage of personal protective equipment (PPE) during the COVID-19 pandemic, the interest and demand for sterilization devices to reuse PPE has increased. For reuse of face masks, they must be effectively decontaminated of potential infectious agents without compromising its filtration ability during sterilization. In this study, we utilized an atmospheric pressure pulsed dielectric barrier discharge (DBD), combined with nebulized liquid microdroplets to generate plasma-activated mist (PAM). MS2 and T4 bacteriophages were used to conduct the decontamination tests on two types of N95 respirators. Results showed at least a 2-log reduction of MS2 and T4 on N95 respirators treated in one cycle with 7.8% hydrogen peroxide PAM and at least a 3-log reduction treated in 10% hydrogen peroxide PAM. In addition, it was found that there was no significant degradation in filtration efficiency of N95 respirators (3M 1860 and 1804) treated in 10% hydrogen peroxide PAM found after 20 cycles. In terms of re-useability of masks after treatment as determined, it was shown that the elastic straps of 3M 1804 were fragmented after 20 treatment cycles rendering them unusable, while the straps of 3M 1860 were not negatively affected even after 20 disinfection cycles.

Exposure pathways and bioaccumulation of per- and polyfluoroalkyl substances in freshwater aquatic ecosystems: Key considerations.

Due to the bioaccumulative behavior, toxicity, and recalcitrance to degradation, per- and polyfluoroalkyl substances (PFAS) are a focus for many researchers investigating freshwater aquatic ecosystems. PFAS are a diverse set of chemicals that accumulate and transport quite differently in the environment depending on the length of their fluoroalkyl chains and their functional groups. This diversity in PFAS chemical characteristics combined with varying environmental factors also impact the bioaccumulation of these compounds in different organisms. In this review, we evaluate environmental factors (such as organic carbon, proteins, lipids, and dissolved cations) as well as PFAS characteristics (head group, chain-length, and concentration) that contribute to the significant variation seen in the literature of bioaccumulation metrics reported for organisms in aquatic ecosystems. Of the factors evaluated, it was found that PFAS concentration, dissolved organic matter, sediment organic matter, and biotransformation of precursor PFAS tended to significantly impact reported bioaccumulation metrics the most. Based on this review, it is highly suggested that future studies provide sufficient details of important environmental factors, specific organism traits/ behavior, and PFAS concentrations/compounds when reporting on bioaccumulation metrics to further fill data gaps and improve our understanding of PFAS in aquatic ecosystems.

Linking PFAS partitioning behavior in sewage solids to the solid characteristics, solution chemistry, and treatment processes.

Per- and polyfluoroalkyl substances (PFAS) have gained increasing attention due to the potential health risks that they present. Secondary sludge and biosolids are known as notable PFAS emission routes to the environment. In this study, partitioning behavior of 14 PFAS were investigated across four secondary wastewater treatment types (activated sludge, trickling filter, biological nutrient removal, and rotating biological contactor; n = 10) and three sludge stabilization methods (composting, aerobic digestion, and anaerobic digestion; n = 6). Batch experiments were conducted to evaluate how PFAS sorption to secondary sludge and biosolid was affected by various treatment methods, solid properties, and solution chemistry parameters. Insignificant differences in compound-specific partitioning coefficients (Kd) were observed among the four secondary treatment methods. However, sludge stabilization resulted in significantly different partitioning behavior among biosolid samples, in which anaerobically digested biosolids generally had significantly higher Kd values compared to aerobically digested and composted biosolids (anaerobic digestion > aerobic digestion > composting). Multiple linear regression models were developed to explain analyte-specific Kd values across the biosolid samples and identified that solid-specific property significance was as follows: protein fraction > organic matter fraction > lipid fraction. Stabilization generally decreased the PFAS sorption capacity relative to the secondary sludge samples. Furthermore, PFAS Kd increased with elevated calcium concentrations and ionic strengths and decreased with increasing pH values in sludge and biosolid samples. These findings could inform the decision-making process to reduce the release of PFAS to the environment.

Wastewater treatment plant effluent introduces recoverable shifts in microbial community composition in receiving streams.

Through a combined approach using analytical chemistry, real-time quantitative polymerase chain reaction (qPCR), and targeted amplicon sequencing, we studied the impact of wastewater treatment plant effluent sources at six sites on two sampling dates on the chemical and microbial population regimes within the Wissahickon Creek, and its tributary, Sandy Run, in Montgomery County, Pennsylvania, USA. These water bodies contribute flow to the Schuylkill River, one of the major drinking water sources for Philadelphia, Pennsylvania. Effluent was observed to be a significant source of nutrients, human and non-specific fecal associated taxa. There was an observed increase in the alpha diversity at locations immediately below effluent outflows, which contributed many taxa involved in wastewater treatment processes and nutrient cycling to the stream's microbial community. Unexpectedly, modeling of microbial community shifts along the stream was not controlled by concentrations of measured nutrients. Furthermore, partial recovery, in the form of decreasing abundances of bacteria and nutrients associated with wastewater treatment plant processes, nutrient cycling bacteria, and taxa associated with fecal and sewage sources, was observed between effluent sources, which we hypothesize is controlled by distance from effluent source. Antecedent moisture conditions were observed to impact overall microbial community diversity, with higher diversity occurring after rainfall. Finally, the efficacy of using a subset of the microbial community including the orders of Bifidobacteriales, Bacteroidales, and Clostridiales to estimate the degree of influence due to sewage and fecal sources was explored and verified.

Influence of Scale on Biomass Growth and Nutrient Removal in an Algal-Bacterial Leachate Treatment System.

Data collected from experiments conducted at a flask scale are regularly used as input data for life cycle assessments and techno-economic analyses for predicting the potential productivities of large-scale commercial facilities. This study measures and compares nitrogen removal and biomass growth rates in treatment systems that utilize an algae-bacteria consortium to remediate landfill leachate at three scales: small (0.25 L), medium (100 L), and large (1000 L). The medium- and large-scale vessels were run for 52 consecutive weeks as semibatch reactors under variable environmental conditions. The small-scale experiments were conducted in flasks as batch experiments under controlled environmental conditions. Kolomogov-Smirnov statistical tests, which compare the distributions of entire data sets, were used to determine if the ammonia removal, total nitrogen removal, and biomass growth rates at each scale were statistically different. Results from the Kolmogov-Smirnov comparison indicate that there is a significant difference between all rates determined in the large-scale vessels compared to those in the small-scale vessels. These results suggest that small-scale experiments may not be appropriate as input data in predictive analyses of full scale algal processes. The accumulation of nitrite and nitrate within the reactor, observed midway through the experimental process, is attributed to high relative abundances of ammonia- and nitrite-oxidizing bacteria, identified via metagenomic analysis.

Comparison of Scale in a Photosynthetic Reactor System for Algal Remediation of Wastewater.

An experimental methodology is presented to compare the performance of two different sized reactors designed for wastewater treatment. In this study, ammonia removal, nitrogen removal and algal growth are compared over an 8-week period in paired sets of small (100 L) and large (1,000 L) reactors designed for algal remediation of landfill wastewater. Contents of the small and large scale reactors were mixed before the beginning of each weekly testing interval to maintain equivalent initial conditions across the two scales. System characteristics, including surface area to volume ratio, retention time, biomass density, and wastewater feed concentrations, can be adjusted to better equalize conditions occurring at both scales. During the short 8-week representative time period, starting ammonia and total nitrogen concentrations ranged from 3.1-14 mg NH3-N/L, and 8.1-20.1 mg N/L, respectively. The performance of the treatment system was evaluated based on its ability to remove ammonia and total nitrogen and to produce algal biomass. Mean ± standard deviation of ammonia removal, total nitrogen removal and biomass growth rates were 0.95±0.3 mg NH3-N/L/day, 0.89±0.3 mg N/L/day, and 0.02±0.03 g biomass/L/day, respectively. All vessels showed a positive relationship between the initial ammonia concentration and ammonia removal rate (R2=0.76). Comparison of process efficiencies and production values measured in reactors of different scale may be useful in determining if lab-scale experimental data is appropriate for prediction of commercial-scale production values.

Fructose Accelerates UV-C Induced Photochemical Degradation of Pentachlorophenol in Low and High Salinity Water.

A novel process involving 254 nm UV-C and fructose to degrade pentachlorophenol (PCP), a pollutant, in low and high salinity (0-10 g/L salt) solutions is presented. The first order rate constants in the presence of 0, 300, and 500 mM fructose were 0.23 ± 0.04, 0.54 ± 0.01, and 1.18 ± 0.03 min(-1), respectively. Experimental evidence has shown generation of hydrogen peroxide and singlet oxygen from the UV-C exposure of fructose, which may have accelerated PCP degradation. Although salts (sodium, potassium, and calcium chloride, 1101:6.4:1) are expected to enhance the degradation rate due to generation of reactive halide species (RHS) from exposure to UV-C light, 10 g/L salt decreased the degradation rates in both the absence and presence of fructose. An LC-ESI-MS spectrum of the reaction mixture revealed a high relative abundance at m/z of 215 that corresponds to a fructose-chlorine adduct, indicating that fructose may have scavenged these RHS and prevented their reaction with PCP.

Nitrogen removal from raw landfill leachate by an algae-bacteria consortium.

A remediation system for the removal of nitrogen from landfill leachate by a mixed algae-bacteria culture was investigated. This system was designed to treat leachate with minimal inputs and maintenance requirements, and was operated as an open semi-batch reactor in an urban greenhouse. The results of this study showed a maximum nitrogen removal rate of 9.18 mg N/(L·day) and maximum biomass density of 480 mg biomass/L. The ammonia removal rates of this culture increased with increasing initial ammonia concentration; maximum nitrogen removal occurred at an ammonia concentration of 80 mg N-NH3/L. At starting ammonia concentrations above 80 mg N-NH3/L a reduction in nitrogen removal was seen; this inhibition is hypothesized to be caused by ammonia toxicity. This inhibiting concentration is considerably higher than that of many other published studies.

Fructose as a novel photosensitizer: Characterization of reactive oxygen species and an application in degradation of diuron and chlorpyrifos.

The objective of this study was to identify reactive oxygen species (ROS) generated from the exposure of fructose solution to the 254 nm ultraviolet (UV) light and evaluate whether fructose can be used as a photosensitizer for accelerated photo-degradation of diuron and chlorpyrifos. We demonstrated that hydrogen peroxide, singlet oxygen ((1)O2) and acidic photolysis products were generated upon UV exposure of fructose. Consistent with these findings, UV induced degradation of chlorpyrifos and diuron was accelerated by the presence of 500 mM fructose. The average first order photo-degradation rate constants in the absence and presence of 500 mM fructose were 0.92 and 2.07 min(-1) respectively for diuron and 0.04 and 0.07 min(-1) for chlorpyrifos. The quantum yields (ɸ) for direct photo-degradation of diuron and chlorpyrifos were 0.003 and 0.001 respectively. In the presence of 500 mM fructose, these values increased to 0.006 and 0.002 respectively. Thus, fructose may be an effective photosensitizer.

Resource recovery from wastewater: application of meta-omics to phosphorus and carbon management.

A growing trend at wastewater treatment plants is the recovery of resources and energy from wastewater. Enhanced biological phosphorus removal and anaerobic digestion are two established biotechnology approaches for the recovery of phosphorus and carbon, respectively. Meta-omics approaches (meta-genomics, transcriptomics, proteomics, and metabolomics) are providing novel biological insights into these complex biological systems. In particular, genome-centric metagenomics analyses are revealing the function and physiology of individual community members. Querying transcripts, proteins and metabolites are emerging techniques that can inform the cellular responses under different conditions. Overall, meta-omics approaches are shedding light into complex microbial communities once regarded as 'blackboxes', but challenges remain to integrate information from meta-omics into engineering design and operation guidelines.

A Novel Bioreactor for High Density Cultivation of Diverse Microbial Communities.

A novel reactor design, coined a high density bioreactor (HDBR), is presented for the cultivation and study of high density microbial communities. Past studies have evaluated the performance of the reactor for the removal of COD(1) and nitrogen species(2-4) by heterotrophic and chemoautotrophic bacteria, respectively. The HDBR design eliminates the requirement for external flocculation/sedimentation processes while still yielding effluent containing low suspended solids. In this study, the HDBR is applied as a photobioreactor (PBR) in order to characterize the nitrogen removal characteristics of an algae-based photosynthetic microbial community. As previously reported for this HDBR design, a stable biomass zone was established with a clear delineation between the biologically active portion of the reactor and the recycling reactor fluid, which resulted in a low suspended solid effluent. The algal community in the HDBR was observed to remove 18.4% of total nitrogen species in the influent. Varying NH4(+) and NO3(-) concentrations in the feed did not have an effect on NH4(+) removal (n=44, p=0.993 and n=44, p=0.610 respectively) while NH4(+) feed concentration was found to be negatively related with NO3(-) removal (n=44, p=0.000) and NO3(-) feed concentration was found to be positively correlated with NO3(-) removal (n=44, p=0.000). Consistent removal of NH4(+), combined with the accumulation of oxidized nitrogen species at high NH4(+) fluxes indicates the presence of ammonia- and nitrite-oxidizing bacteria within the microbial community.

Simultaneous autotrophic denitrification and nitrification in a low-oxygen reaction environment.

The occurrence of autotrophic denitrification and nitrification activities by ammonia-oxidising bacteria and nitrite-oxidising bacteria is studied in a bioreactor system operable at low-dissolved oxygen (DO) and at variable oxygen influx rates. At a loading of 3.6 mg NH4(+)-N/h into the bioreactor, simultaneous autotrophic denitrification and nitrification contributed to NH4(+)-N removal over oxygen influxes of 2-14 mg O2/h and DO <0.5 mg/L. The maximum autotrophic denitrification (or total-N removal) rates were achieved in a narrow oxygen influx band of 3-5 mg O2/h, where it accounted for up to 36% of NH4(+)-N removal. At oxygen influx >16 mg O2/h and DO >2 mg/L, autotrophic denitrification ceases and roughly 90% of feed NH4(+)-N is oxidised to NOX(-)-N. The stability of total effluent chemical oxygen demand (COD) over the range of oxygen influxes tested confirms the absence of heterotrophic denitrification in the bioreactor. The long solids residence time of the stable biomass zone (21 days) led to production of effluent COD as a result of cell decay, and thus effluent COD was used to calculate more accurately the mean cell residence time.

Oxidation of the cyclic ethers 1,4-dioxane and tetrahydrofuran by a monooxygenase in two Pseudonocardia species.

The bacterium Pseudonocardia dioxanivorans CB1190 grows on the cyclic ethers 1,4-dioxane (dioxane) and tetrahydrofuran (THF) as sole carbon and energy sources. Prior transcriptional studies indicated that an annotated THF monooxygenase (THF MO) gene cluster, thmADBC, located on a plasmid in CB1190 is upregulated during growth on dioxane. In this work, transcriptional analysis demonstrates that upregulation of thmADBC occurs during growth on the dioxane metabolite ß-hydroxyethoxyacetic acid (HEAA) and on THF. Comparison of the transcriptomes of CB1190 grown on THF and succinate (an intermediate of THF degradation) permitted the identification of other genes involved in THF metabolism. Dioxane and THF oxidation activity of the THF MO was verified in Rhodococcus jostii RHA1 cells heterologously expressing the CB1190 thmADBC gene cluster. Interestingly, these thmADBC expression clones accumulated HEAA as a dead-end product of dioxane transformation, indicating that despite its genes being transcriptionally upregulated during growth on HEAA, the THF MO enzyme is not responsible for degradation of HEAA in CB1190. Similar activities were also observed in RHA1 cells heterologously expressing the thmADBC gene cluster from Pseudonocardia tetrahydrofuranoxydans K1.

Glyoxylate metabolism is a key feature of the metabolic degradation of 1,4-dioxane by Pseudonocardia dioxanivorans strain CB1190.

The groundwater contaminant 1,4-dioxane (dioxane) is transformed by several monooxygenase-expressing microorganisms, but only a few of these, including Pseudonocardia dioxanivorans strain CB1190, can metabolize the compound as a sole carbon and energy source. However, nothing is yet known about the genetic basis of dioxane metabolism. In this study, we used a microarray to study differential expression of genes in strain CB1190 grown on dioxane, glycolate (a previously identified intermediate of dioxane degradation), or pyruvate. Of eight multicomponent monooxygenase gene clusters carried by the strain CB1190 genome, only the monooxygenase gene cluster located on plasmid pPSED02 was upregulated with dioxane relative to pyruvate. Plasmid-borne genes for putative aldehyde dehydrogenases, an aldehyde reductase, and an alcohol oxidoreductase were also induced during growth with dioxane. With both dioxane and glycolate, a chromosomal gene cluster encoding a putative glycolate oxidase was upregulated, as were chromosomal genes related to glyoxylate metabolism through the glyoxylate carboligase pathway. Glyoxylate carboligase activity in cell extracts from cells pregrown with dioxane and in Rhodococcus jostii strain RHA1 cells expressing the putative strain CB1190 glyoxylate carboligase gene further demonstrated the role of glyoxylate metabolism in the degradation of dioxane. Finally, we used (13)C-labeled dioxane amino acid isotopomer analysis to provide additional evidence that metabolites of dioxane enter central metabolism as three-carbon compounds, likely as phosphoglycerate. The routing of dioxane metabolites via the glyoxylate carboligase pathway helps to explain how dioxane is metabolized as a sole carbon and energy source for strain CB1190.

Genome sequence of the 1,4-dioxane-degrading Pseudonocardia dioxanivorans strain CB1190.

Pseudonocardia dioxanivorans CB1190 is the first bacterium reported to be capable of growth on the environmental contaminant 1,4-dioxane and the first member of the genus Pseudonocardia for which there is an annotated genome sequence. Preliminary analysis of the genome (chromosome and three plasmids) indicates that strain CB1190 possesses several multicomponent monooxygenases that could be involved in the aerobic degradation of 1,4-dioxane and other environmental contaminants.