The sewer environment is a potential hotspot for the proliferation of antibiotic resistance genes (ARGs) and other hazardous microbial agents.

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Combined sewer overflows (CSOs) degrade water quality through the release of microbial contaminants in CSO effluent. Improved understanding of the partitioning of microbial contaminants onto settleable particles can provide insight into their fate in end-of-pipe treatment systems or following release during CSO events. Sampling was performed across the hydrograph for three storm events as well as during baseflow and wet weather in three surface waters impacted by CSO. qPCR was performed for select antibiotic resistance genes (ARG) and a marker gene for human fecal indicator organisms (BacHum) in samples processed the partitioning of microbial contaminants on settleable particles versus suspended in the aqueous phase. Amplicon sequencing was performed on both fractions of storm samples to further define the timing and partitioning of microbial contaminants released during CSO events. Samples collected at the CSO outfall exhibited microbial community signatures of wastewater at select time points early or late in the storm events. CSOs were found to be a source of ARG. In surrounding surface waters, sul1 was higher in samples from select locations during wet weather compared to baseflow. Otherwise, ARG concentrations were variable with no differences between baseflow and wet weather conditions. The majority of ARG at the CSO outfall were observed on the attached fraction of samples: 64–79% of sul1 and 59–88% of tet(G). However, the timing of peak ARG and human fecal indicator marker gene BacHum did not necessarily coincide with observation of the microbial signature of wastewater in CSO effluent. Therefore, unit processes that remove settleable particles (e.g., hydrodynamic separators) operated throughout a CSO event would achieve up to (0.5–0.9)-log removal of ARG and fecal indicators by removing the attached fraction of measured genes. Secondary treatment would be required if greater removal of these targets is needed.

Wastewater systems are recognized pathways for the spread of antibiotic resistant bacteria, but relatively little is known about the microbial ecology of the sewer environment.

Combined sewer overflows (CSOs) degrade water quality and end-of-pipe treatment is one potential solution for retrofitting this outdated infrastructure. The goal of this research was to evaluate peracetic acid (PAA) as a disinfectant for CSOs using viability based molecular methods for antibiotic resistance genes (ARGs), indicator organism marker gene BacHum, and 16S rRNA genes. Simulated CSO effluent was prepared using 23–40% wastewater, representing the higher end of the range of wastewater concentrations reported in CSO effluent. PAA residual following disinfection was greatest for samples with the lowest initial COD. Treatment of simulated CSO effluent (23% wastewater) with 100 mg∙min/L PAA (5 mg/L PAA, 20 min) was needed to reduce viable cell sul1, tet(G), and BacHum (1.0±0.63–3.2±0.25-log) while 25 to 50 mg•min/L PAA (5 mg/L PAA, 5–10 min) was needed to reduce viable cell loads (0.62±0.56–1.6±0.08-log) in 40% wastewater from a different municipal treatment plant. Increasing contact time after the initial decrease in viable cell gene copies did not significantly improve treatment. A much greater applied Ct of 1200 mg∙min/L PAA (20 mg/L PAA, 60 min) was required for significant log reduction of 16S rRNA genes (3.29±0.13-log). No significant losses of mexB were observed during the study. Data were fitted to a Chick-Watson model and resulting inactivation constants for sul1 and tet(G) > BacHum > 16S rRNA. Amplicon sequencing of the 16S rRNA gene indicated the initial viable and total microbial communities were distinct and that treatment with PAA resulted in marked increases of the relative abundance of select phyla, particularly Clostridia which increased by 1–1.5 orders of magnitude. Results confirm that membrane disruption is a mechanism for PAA disinfection and further treatment is needed to reduce total ARGs in CSO effluent.

Antibiotic resistance is a public health issue with links to environmental sources of antibiotic resistance genes (ARGs). ARGs from nonviable sources may pose a hazard given the potential for transformation whereas ARGs in viable sources may proliferate during host growth or conjugation. In this study, ARGs in the effluent from three municipal wastewater treatment plants (WWTPs) and the receiving surface waters were investigated using a viability-based qPCR technique (vPCR) with propidium monoazide (PMA). ARGs sul1, tet(G), and bla TEM , fecal indicator marker BacHum, and 16S rRNA gene copies/mL were found to be significantly lower in viable-cells than in total concentrations for WWTP effluent. Viable-cell and total gene copy concentrations were similar in downstream samples except for tet(G). Differences with respect to season in the prevalence of nonviable ARGs in surface water or WWTP effluent were not observed. The results of this study indicate that qPCR may overestimate viable-cell ARGs and fecal indicator genes in WWTP effluent but not necessarily in the surface water >1.8km downstream.

A series of microcosm experiments were performed to determine the effectiveness of various gaseous electron donors (including hydrogen, 1‐hexene, ethyl acetate, and liquefied petroleum gas [LPG]) for supporting biological perchlorate reduction under different electron donor concentrations and soil moistures. Under high soil moisture (16% w/w) conditions, complete or partial perchlorate degradation was achieved with all of the tested electron donors, except for ethyl acetate. Hydrogen was the most promising of the tested electron donors, achieving complete perchlorate degradation with first‐order rate constants ranging from 0.13 to 0.20 day−1 and reducing concentrations to non‐detectable levels within 35 to 42 days. The LPG and 1‐hexene each promoted partial perchlorate reduction, with average first‐order rate constants of 0.05 and 0.11 day−1, respectively. Although significant perchlorate reduction was observed with as little as 13% moisture, the moisture content for complete perchlorate degradation in this particular soil was determined to be 17%.

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