Correlation of crAssphage qPCR Markers with Culturable and Molecular Indicators of Human Fecal Pollution in an Impacted Urban Watershed.

Environmental waters are monitored for fecal pollution to protect public health. Many previously developed human-specific fecal pollution indicators lack adequate sensitivity to be reliably detected in environmental waters or do not correlate well with viral pathogens. Recently, two novel human sewage-associated source tracking qPCR markers were developed based on the bacteriophage crAssphage, CPQ_056 and CPQ_064. These assays are highly human specific, abundant in sewage, and are viral-based, suggesting great promise for environmental application as human fecal pollution indicators. A 30-day sampling study was conducted in an urban stream impacted by combined sewer overflows to evaluate the crAssphage markers' performance in an environmental system. The crAssphage markers were present at concentrations of 4.02-6.04 log10 copies/100 mL throughout the study period, indicating their high abundance and ease of detection in polluted environmental waters. In addition, the crAssphage assays were correlated with rain events, molecular markers for human polyomavirus and HF183, as well as culturable E. coli, enterococci, and somatic coliphage. The CPQ_064 assay correlated strongly to a greater number of biological indicators than the CPQ_056 assay. This study is the first to evaluate both crAssphage qPCR assays in an extended environmental application of crAssphage markers for monitoring of environmental waters. It is also the first study to compare crAssphage marker concentration with other viral-based indicators.

Microbial mats as a biological treatment approach for saline wastewaters: the case of produced water from hydraulic fracturing.

Treatment of produced water, i.e. wastewater from hydraulic fracturing, for reuse or final disposal is challenged by both high salinity and the presence of organic compounds. Organic compounds in produced water may foul physical-chemical treatment processes or support microbial corrosion, fouling, and sulfide release. Biological approaches have potential applications in produced water treatment, including reducing fouling of physical-chemical treatment processes and decreasing biological activity during produced water holding; however, conventional activated sludge treatments are intolerant of high salinity. In this study, a biofilm treatment approach using constructed microbial mats was evaluated for biodegradation performance, microbial community structure, and metabolic potential in both simulated and real produced water. Results demonstrated that engineered microbial mats are active at total dissolved solids (TDS) concentrations up to at least 100,000 mg/L, and experiments in real produced water showed a biodegradation capacity of 1.45 mg COD/gramwet-day at a TDS concentration of 91,351 mg/L. Additionally, microbial community and metagenomic analyses revealed an adaptive microbial community that shifted based upon the sample being treated and has the metabolic potential to degrade a wide array of contaminants, suggesting the potential of this approach to treat produced waters with varying composition.

Mainstream partial denitrification-anammox in sand and expanded clay deep-bed polishing filters under practical loading rates and backwashing conditions.

This study focused on evaluating the feasibility of expanded clay and sand as media types for mainstream partial denitrification-anammox (PdNA) in deep-bed single-media polishing filters under nitrogen and solids loading rates as well as backwash conditions similar to conventional denitrification filters. The surface roughness and iron content of the expanded clay were hypothesized to allow for enhanced anammox retention, nitrogen removal rates, and runtimes. However, under the tested loading rates and backwash conditions, no clear benefit of expanded clay was observed compared with conventional sand. This study showed the feasibility of PdNA in filters with both sand and expanded clay with PdN efficiencies of 76% and 77%, PdNA rates of 840 and 843 g N/m3 /d and TIN removal rates of 960 and 964 g N/m3 /d, respectively. Glycerol demands were 1.5-1.6 g COD added per g TIN removed , thus indicating potential carbon savings up to 75% compared with conventional denitrification. Overall, this study showed for the first time PdNA filters performing at nitrogen removal rates double that of previous PdNA studies under realistic conditions while providing insights into the media choice and backwashing conditions. Future research on expanded clay backwash conditions is needed to provide its full potential in PdNA filters. PRACTITIONER POINTS: Hydraulic and TSS loading rates similar to conventional denitrification can be applied in PdNA filters. Conventional sand can be used when retrofitting conventional denitrification filters into PdNA filters. Carbon savings up to 75% can be achieved with glycerol when retrofitting conventional filters into PdNA filters.

Glutaraldehyde inhibits biological treatment of organic additives in hydraulic fracturing produced water.

Water generated by hydraulic fracturing for the production of oil and gas, commonly termed 'produced water', may contain residual organic compounds from the fracturing process or the subsurface formation. Biological treatment is a potential technology to remove residual organic compounds in produced water. Biocides are often added to both fracturing fluids and produced water to limit undesirable microbiological activity, and glutaraldehyde is the most commonly used biocide in hydraulic fracturing. Residual biocides in produced water can limit biological treatment efficiency. We evaluated the effect of glutaraldehyde on the biodegradation of five of the most commonly reported organic compounds in hydraulic fracturing fluids in an engineered biofilm treatment. Our results demonstrated that glutaraldehyde delays biological organic compound removal by introducing a biodegradation lag phase. In addition, the effects of glutaraldehyde were more pronounced for more rapidly degraded compounds. Finally, the presence of glutaraldehyde did not decrease microbial abundance nor drive microbial community structure, suggesting that observed effects were due to altered microbial activity. These results highlight the necessity to consider co-contaminant interactions during treatment of complex waste streams where residual biocide may be present.

Characterization and biological removal of organic compounds from hydraulic fracturing produced water.

Hydraulic fracturing generates large volumes of produced water, and treatment of produced water may be necessary for disposal or reuse. Biological treatment of produced water is a potential approach to remove organic constituents and reduce fouling, in conjunction with other treatment processes. This study investigates the biological treatability of produced water samples from the Utica and Bakken Shales using engineered biofilms. Observed total dissolved organic carbon (DOC) removal varied between 1-87% at normalized total dissolved solids concentrations, suggesting that the composition of produced water, including organic constituents and trace elements such as nutrients and metals, is an important driver of biological treatment performance. Mass spectrometric analyses of the DOC composition revealed various alkanes in all samples, but differences in non-ionic surfactant, halogenated, and acidic compound content. Statistical data reduction approaches suggest that the latter two groups are correlated with reduced biodegradation kinetics. These results demonstrate that the combination of biodegradation performance and organic speciation can guide the assessment of the biological treatment of produced water.