Enhanced bioremediation of RDX and Co-Contaminants perchlorate and nitrate using an anaerobic dehalogenating consortium in a fractured rock aquifer.

The potential neurotoxic and carcinogenic effects of the explosives compound RDX (hexahydro-1,3,5-trinitro-1,3,5-triazine) on human health requires groundwater remediation strategies to meet low cleanup goals. Bioremediation of RDX is feasible through biostimulation of native microbes with an organic carbon donor but may be less efficient, or not occur at all, in the presence of the common co-contaminants perchlorate and nitrate. Laboratory tests compared biostimulation with bioaugmentation to achieve anaerobic degradation of RDX, perchlorate, and nitrate; a field pilot test was then conducted in a fractured rock aquifer with the selected bioaugmentation approach. Insignificant reduction of RDX, perchlorate, or nitrate was observed by the native microbes in microcosms, with or without biostimulation by addition of lactate. Tests of the RDX-degrading ability of the microbial consortium WBC-2, originally developed for dehalogenation of chlorinated volatile organic compounds, showed first-order biodegradation rate constants ranging from 0.57 to 0.90 per day (half-lives 1.2 to 0.80 days). WBC-2 sustained degradation without daughter product accumulation when repeatedly amended with RDX and lactate for a year. In microcosms with groundwater containing perchlorate and nitrate, RDX degradation began without delay when bioaugmented with 10% WBC-2. Slower RDX degradation occurred with 3% or 5% WBC-2 amendment, indicating a direct relation with cell density. Transient RDX daughter compounds included methylene dinitramine, MNX, and DNX. With WBC-2 amendment, nitrate concentrations immediately decreased to near or below detection, and perchlorate degradation occurred with half-lives of 25-34 days. Single-well injection tests with WBC-2 and lactate showed that the onset of RDX degradation coincided with the onset of sulfide production, which was affected by the initial perchlorate concentration. Biodegradation rates in the pilot injection tests agreed well with those measured in the microcosms. These results support bioaugmentation with an anaerobic culture as a remedial strategy for sites contaminated with RDX, nitrate, and perchlorate.

Predicting COVID-19 Infected Individuals in a Defined Population from Wastewater RNA Data.

Wastewater surveillance of SARS-CoV-2 RNA has become an important tool for tracking the presence of the virus and serving as an early indicator for the onset of rapid transmission. Nevertheless, wastewater data are still not commonly used to predict the number of infected individuals in a sewershed. The main objective of this study was to calibrate a susceptible-exposed-infectious-recovered (SEIR) model using RNA copy rates in sewage (i.e., gene copies per liter times flow rate) and the number of SARS-CoV-2 saliva-test-positive infected individuals in a university student population that was subject to repeated weekly testing during the Spring 2021 semester. A strong correlation was observed between the RNA copy rates and the number of infected individuals. The parameter in the SEIR model that had the largest impact on calibration was the maximum shedding rate, resulting in a mean value of 7.72 log10 genome copies per gram of feces. Regressing the saliva-test-positive infected individuals on predictions from the SEIR model based on the RNA copy rates yielded a slope of 0.87 (SE=0.11), which is statistically consistent with a 1:1 relationship between the two. These findings demonstrate that wastewater surveillance of SARS-CoV-2 can be used to estimate the number of infected individuals in a sewershed.

COVID-19 wastewater epidemiology: a model to estimate infected populations.

BACKGROUND: Wastewater-based epidemiology provides an opportunity for near real-time, cost-effective monitoring of community-level transmission of SARS-CoV-2. Detection of SARS-CoV-2 RNA in wastewater can identify the presence of COVID-19 in the community, but methods for estimating the numbers of infected individuals on the basis of wastewater RNA concentrations are inadequate. METHODS: This is a wastewater-based epidemiology study using wastewater samples that were collected weekly or twice a week from three sewersheds in South Carolina, USA, between either May 27 or June 16, 2020, and Aug 25, 2020, and tested for SARS-CoV-2 RNA. We developed a susceptible-exposed-infectious-recovered (SEIR) model based on the mass rate of SARS-CoV-2 RNA in the wastewater to predict the number of infected individuals, and have also provided a simplified equation to predict this. Model predictions were compared with the number of confirmed cases identified by the Department of Health and Environmental Control, South Carolina, USA, for the same time period and geographical area. FINDINGS: We plotted the model predictions for the relationship between mass rate of virus release and numbers of infected individuals, and we validated this prediction on the basis of estimated prevalence from individual testing. A simplified equation to estimate the number of infected individuals fell within the 95% confidence limits of the model. The rate of unreported COVID-19 cases, as estimated by the model, was approximately 11 times that of confirmed cases (ie, ratio of estimated infections for every confirmed case of 10·9, 95% CI 4·2-17·5). This rate aligned well with an independent estimate of 15 infections for every confirmed case in the US state of South Carolina. INTERPRETATION: The SEIR model provides a robust method to estimate the total number of infected individuals in a sewershed on the basis of the mass rate of RNA copies released per day. This approach overcomes some of the limitations associated with individual testing campaigns and thereby provides an additional tool that can be used to inform policy decisions. FUNDING: Clemson University, USA.

Biogeochemical oxidation of calcium sulfite hemihydrate to gypsum in flue gas desulfurization byproduct using sulfur-oxidizing bacteria.

Flue gas desulfurization (FGD) is a well-established air treatment technology for coal and oil combustion gases that commonly uses lime or pulverized limestone aqueous slurries to precipitate sulfur dioxide (SO2) as crystalline calcium salts. Under forced oxidation (excess oxygen) conditions, FGD byproduct contains almost entirely (>92%) gypsum (CaSO4·2H2O), a useful and marketable commodity. In contrast, FGD byproduct formed in oxygen deficient oxidation systems contains a high percentage of hannebachite (CaSO3·0.5H2O) to yield a material with no commercial value, poor dewatering characteristics, and that is typically disposed in landfills. Hannebachite in FGD byproduct can be chemically converted to gypsum; however, the conditions that support rapid formation of gypsum require large quantities of acids or oxidizers. This work describes a novel, patent pending application of microbial physiology where a natural consortium of sulfur-oxidizing bacteria (SOB) was used to convert hannebachite-enriched FGD byproduct into a commercially valuable, gypsum-enriched product (US Patent Assignment 503373611). To optimize the conversion of hannebachite into gypsum, physiological studies on the SOB were performed to define their growth characteristics. The SOB were found to be aerobic, mesophilic, neutrophilic, and dependent on a ready supply of ammonia. They were capable of converting hannebachite to gypsum at a rate of approximately five percent per day when the culture was applied to a 20 percent FGD byproduct slurry and SOB growth medium. 16S rDNA sequencing revealed that the SOB consortium contained a variety of different bacterial genera including both SOB and sulfate-reducing bacteria. Halothiobacillus, Thiovirga and Thiomonas were the dominant sulfur-oxidizing genera.

Natural Attenuation in Streambed Sediment Receiving Chlorinated Solvents from Underlying Fracture Networks.

Contaminant discharge from fractured bedrock formations remains a remediation challenge. We applied an integrated approach to assess the natural attenuation potential of sediment that forms the transition zone between upwelling groundwater from a chlorinated solvent-contaminated fractured bedrock aquifer and the receiving surface water. In situ measurements demonstrated that reductive dechlorination in the sediment attenuated chlorinated compounds before reaching the water column. Microcosms established with creek sediment or in situ incubated Bio-Sep beads degraded C1-C3 chlorinated solvents to less-chlorinated or innocuous products. Quantitative PCR and 16S rRNA gene amplicon sequencing revealed the abundance and spatial distribution of known dechlorinator biomarker genes within the creek sediment and demonstrated that multiple dechlorinator populations degrading chlorinated C1-C3 alkanes and alkenes co-inhabit the sediment. Phylogenetic classification of bacterial and archaeal sequences indicated a relatively uniform distribution over spatial (300 m horizontally) scale, but Dehalococcoides and Dehalobacter were more abundant in deeper sediment, where 5.7 ± 0.4 × 105 and 5.4 ± 0.9 × 106 16S rRNA gene copies per g of sediment, respectively, were measured. The microbiological and hydrogeological characterization demonstrated that microbial processes at the fractured bedrock-sediment interface were crucial for preventing contaminants reaching the water column, emphasizing the relevance of this critical zone environment for contaminant attenuation.

Performance of an anaerobic, static bed, fixed film bioreactor for chlorinated solvent treatment.

Anaerobic, fixed film, bioreactors bioaugmented with a dechlorinating microbial consortium were evaluated as a potential technology for cost effective, sustainable, and reliable treatment of mixed chlorinated ethanes and ethenes in groundwater from a large groundwater recovery system. Bench- and pilot-scale testing at about 3 and 13,500 L, respectively, demonstrated that total chlorinated solvent removal to less than the permitted discharge limit of 100 µg/L. Various planned and unexpected upsets, interruptions, and changes demonstrated the robustness and reliability of the bioreactor system, which handled the operational variations with no observable change in performance. Key operating parameters included an adequately long hydraulic retention time for the surface area, a constant supply of electron donor, pH control with a buffer to minimize pH variance, an oxidation reduction potential of approximately -200 millivolts or lower, and a well-adapted biomass capable of degrading the full suite of chlorinated solvents in the groundwater. Results indicated that the current discharge criteria can be met using a bioreactor technology that is less complex and has less downtime than the sorption based technology currently being used to treat the groundwater.