Seasonal variation of macrolide resistance gene abundances in the South Fork Iowa River Watershed

The Midwestern United States is dominated by agricultural production with high concentrations of swine, leading to application of swine manure onto lands with artificial subsurface drainage. Previous reports have indicated elevated levels of antibiotic resistance genes (ARGs) in surface water and groundwater around confined animal feeding operations which administer antimicrobials. While previous studies have examined the occurrence of ARGs around confined swine feeding operations, little information is known how their transport from tile-drained fields receiving swine manure application impacts downstream environments. To further our knowledge in this area, water samples were collected from five locations in the agriculturally dominated South Fork Iowa River Watershed with approximately 840,000 swine present in the 76,000 ha basin. Samples were collected monthly from three stream sites and two main artificial subsurface drainage outlets. Samples were analyzed for macrolide resistance genes ermB, ermF and 16S rRNAgene abundance using qPCR. Abundance of erm genes ranged from below limits of quantification to \u3e 107 copies 100 mL− 1 water. Eighty-nine percent of stream water samples contained one of these two ARGs. Results indicate significantly more ermB and ermF in main drainage outlets than stream samples when normalized by 16S rRNA abundance (p \u3c 0.0001). Both artificial drainage locations revealed temporal trends for ermB and ermF abundance when normalized to 16S rRNA abundance. The higher resistance gene concentrations identified in artificial drainage samples occurring mid-Spring and late-Fall are likely due to manure application

Investigating the dispersal of antibiotic resistance associated genes from manure application to soil and drainage waters in simulated agricultural farmland systems

Manure from animals that have been treated with antibiotics is often used to fertilize agricultural soils and its application has previously been shown to enrich for genes associated with antibiotic resistance in agroecosystems. To investigate the magnitude of this effect, we designed a column experiment simulating manure-treated agricultural soil that utilizes artificial subsurface drainage to determine the duration and extent which this type of manure fertilization impacts the set of genes associated with antibiotic resistance in drainage water. We classified ARGs in manure-treated drainage effluent water by its source of origin. Overall, we found that 61% and 7% of the total abundance of ARGs found in drainage water samples could be attributed to manure enrichment and manure addition, respectively. Among these ARGs, we identified 75 genes unique to manure that persisted in both soil and drainage water throughout a drainage season typical of the Upper Midwestern United States. While most of these genes gradually decreased in abundance over time, the IS6100-associated tet(33) gene accrued. These results demonstrate the influence of manure applications on the composition of the resistome observed in agricultural drainage water and highlight the importance of anthropogenic ARGs in the environment

Electrical stimulation for enhanced denitrification in woodchip bioreactors: Opportunities and challenges

Woodchip bioreactors are being implemented for the removal of nitrates in groundwater and tile water drainage. However, low nitrate removals in denitrifying woodchip bioreactors have been observed for short hydraulic retention time (HRT) and low water temperature (°C). One potential approach to improve woodchip bioreactor performance is to provide an alternative and readily available electron source to the denitrifying microorganisms through electrical stimulation. Previous work has demonstrated the capability of bio-electrochemical reactors (BER) to remove a variety of water contaminants, including nitrate, in the presence of a soluble carbon source. The objective of this study was to evaluate the denitrification efficiency of electrically augmented woodchip bioreactors and conduct a simple techno-economic analysis (TEA) to understand the possibilities and limitations for full-scale BER implementation for treatment of agricultural drainage. Up-flow column woodchip bioreactors were studied included two controls (non-energized, and without electrodes), two electrically enhanced bioreactors, each using a single 316 stainless steel anode coupled with graphite cathodes, and two electrically enhanced bioreactors, each with graphite for both anode and cathodes. Both pairs of electrically enhanced bioreactors demonstrated higher denitrification efficiencies than controls when 500 mA of current was applied. While this technology appeared promising, the techno-economic analysis showed that the normalized N removal cost ($/kg N) for BERs was 2–10 times higher than the base cost with no electrical stimulation. With our current reactor design, opportunities to make this technology cost effective require denitrification efficiency of 85% at 100 mA. This work informs the process and design of electrically stimulated woodchip bioreactors with optimized performance to achieve lower capital and maintenance costs, and thus lower N removal cost

Impact of temperature and hydraulic retention time on pathogen and nutrient removal in woodchip bioreactors

Woodchip denitrification bioreactors are an important edge-of-field practice for treating agricultural drainage; however, their ability to filter microbial pollutants has primarily been explored in the context of wastewater treatment. Upflow column reactors were constructed and tested for E. coli, Salmonella, NO3-N, and dissolved reactive phosphorus (DRP) at hydraulic retention times (HRTs) of 12 and 24 h and at controlled temperatures of 10 and 21.5 °C. Influent solution was spiked to 30 mg L−1 NO3-N, 2–8 × 105 E. coli and Salmonella, and 0.1 mg L−1 DRP. Microbial removal was consistently observed with removal ranging from 75 to 78% reduction at 10 °C and 90–96% at 21.5 °C. The concentration reduction ranged from 2.75 to 9.03 × 104 for both organisms. HRT had less impact on microbial removal than temperature and thus further investigation of removal under lower HRTs is warranted. Nitrate concentrations averaged 96% reduction (with load removal of 14.6 g N m−3 d−1) from 21.5 °C columns at 24 HRT and 29% reduction (with load removal of 8.8 g N m−3 d−1) from 10 °C columns at 12 HRT. DRP removal was likely temporary due to microbial uptake. While potential for removal of E. coli and Salmonella by woodchip bioreactors is demonstrated, system design will need to be considered. High concentrations of these microbial contaminants are likely to occur during peak flows, when bypass flow may be occurring. The results of this study show that woodchip bioreactors operated for nitrate removal have a secondary benefit through the removal of enteric bacteria

In‐situ extraction and impregnation of black walnut husk into polyethylene film using supercritical carbon dioxide with an ethanol modifier

Walnuts are commonly cultivated for their kernel, which is a rich source of antioxidant phenolic compounds. The husk likewise contains antioxidant and antimicrobial compounds, but is typically discarded without further processing. Antioxidant compounds are useful in creating active packaging films, but typically decompose at melt extrusion temperatures in polymer processing. Due to carbon dioxide\u27s low critical point and ability to swell polymer films, supercritical carbon dioxide may be used to impregnate phenolic compounds into polymers. For this study, a novel technique is used to simultaneously produce walnut husk extracts and impregnate the extract into polymer films in the same batch extractor using supercritical carbon dioxide with a 15 wt-% ethanol modifier at 60°C at 19.4 MPa. The effect of varying the loading of walnut husk in the extractor upon impregnation mass was evaluated with the impregnation mass of the film increasing with walnut husk loading. It was determined by FTIR, as well as the eduction of the protein cytochrome c, that antioxidant compounds may be extracted from walnut husks and impregnated into low-density polyethylene film (LDPE) by this technique