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Fecal indicator and pathogenic bacteria and their antibiotic resistance in alluvial groundwater of an irrigated agricultural region with dairies

Surveys of microbiological groundwater quality were conducted in a region with intensive animal agriculture in California, USA. The survey included monitoring and domestic wells in eight concentrated animal feeding operations (CAFOs) and 200 small (domestic and community supply district) supply wells across the region. Campylobacter was not detected in groundwater, whereas Escherichia coli O157:H7 and Salmonella were each detected in 2 of 190 CAFO monitoring well samples. Nonpathogenic generic E. coli and Enterococcus spp. were detected in 24.2% (46/190) and 97.4% (185/190) groundwater samples from CAFO monitoring wells and in 4.2% (1/24) and 87.5% (21/24) of CAFO domestic wells, respectively. Concentrations of both generic E. coli and Enterococcus spp. were significantly associated with well depth, season, and the type of adjacent land use in the CAFO. No pathogenic bacteria were detected in groundwater from 200 small supply wells in the extended survey. However, 4.5 to 10.3% groundwater samples were positive for generic E. coli and Enterococcus. Concentrations of generic E. coli were not significantly associated with any factors, but concentrations of Enterococcus were significantly associated with proximity to CAFOs, seasons, and concentrations of potassium in water. Among a subset of E. coli and Enterococcus isolates from both surveys, the majority of E. coli (63.6%) and Enterococcus (86.1%) isolates exhibited resistance to multiple (≥3) antibiotics. Findings confirm significant microbial and antibiotic resistance loading to CAFO groundwater. Results also demonstrate significant attenuative capacity of the unconfined alluvial aquifer system with respect to microbial transport.

California

Work plan for determining the occurrence of glyphosate, its transformation product AMPA, other herbicide compounds, and antibiotics in midwestern United States streams, 2002

Changes in herbicide use in the Midwestern United States have been substantial over the last 5 years. Most significant is a tripling in the use of glyphosate (N-[phosphonomethyl]glycin). Over this same time period (1997­2001), atrazine use increased by 20 percent and acetochlor use increased by 10 percent, while cyanazine use decreased by 99 percent, alachlor use decreased by 70 percent, and metolachlor use decreased by 55 percent. Previous studies have documented that herbicide flushes occur in midwestern streams during runoff events for several weeks to several months following application, and that herbicide concentrations in midwestern streams during flushing events are related to rates of herbicide use. The objective of this study is to determine the distribution of glyphosate and its primary transformation product aminomethylphosphonic acid (AMPA) in midwestern streams during post-application and harvest-season runoff events. Water samples will be collected in 2002 during two post-herbicide-application runoff events and one harvest-season runoff event from 53 sites on streams in the Midwestern United States. All samples will be analyzed at the U.S. Geological Survey Organic Geochemistry Research Laboratory in Lawrence, Kansas, for glyphosate and 20 other herbicides. Samples will also be analyzed for a glyphosate transformation product (AMPA) and 26 other herbicide transformation products, using GC/MS or HPLC/MS. Selected samples will be analyzed for 36 antibiotics or antibiotic transformational products. Results from this study will represent the first broad-scale investigation of glyphosate and AMPA in U.S. water resources.

Midwest region

Chapter A5. Section 6.1.F. Wastewater, Pharmaceutical, and Antibiotic Compounds

The USGS differentiates between samples collected for analysis of wastewater compounds and those collected for analysis of pharmaceutical and antibiotic compounds, based on the analytical schedule for the laboratory method. Currently, only the wastewater laboratory method for field-filtered samples (SH1433) is an approved, routine (production) method. (The unfiltered wastewater method LC 8033 also is available but requires a proposal for custom analysis.) At this time, analysis of samples for pharmaceutical and antibiotic compounds is confined to research studies and is available only on a custom basis.

Techniques of Water-Resources Investigations

Antibiotic resistance in marine microbial communities proximal to a Florida sewage outfall system

Water samples were collected at several wastewater treatment plants in southeast Florida, and water and sediment samples were collected along and around one outfall pipe, as well as along several transects extending both north and south of the respective outfall outlet. Two sets of samples were collected to address potential seasonal differences, including 38 in the wet season (June 2018) and 42 in the dry season (March 2019). Samples were screened for the presence/absence of 15 select antibiotic resistance gene targets using the polymerase chain reaction. A contrast between seasons was found, with a higher frequency of detections occurring in the wet season and fewer during the dry season. These data illustrate an anthropogenic influence on offshore microbial genetics and seasonal flux regarding associated health risks to recreational users and the regional ecosystem.

Florida

Vegetative buffer strips show limited effectiveness for reducing antibiotic transport in surface runoff

Vegetative buffer strips (VBS) have been demonstrated to effectively reduce loads of sediment, nutrients, and herbicides in surface runoff, but their effectiveness for reducing veterinary antibiotic (VA) loads in runoff has not been well documented. The objective of this study was to determine the effectiveness of VBS vegetation and width on surface runoff loads of the VAs sulfamethazine (SMZ) and lincomycin (LIN). Experimental design of the plots (1.5 × 25 m) was a two-way factorial with four vegetation treatments (tall fescue [ Festuca aruninacea Schreb.]; tall fescue with switchgrass [ Panicum virgatum L.] hedge; warm-season native grass mix; and continuous fallow control), and four buffer widths (0, 2, 5, and 9 m). Turkey litter spiked with SMZ and LIN was applied to the source area (upper 7 m) of each plot, and runoff was collected at each width. Runoff was generated with a rotating boom simulator. Results showed VA loads in runoff at the 0-m sampler ranged from 3.8 to 5.9% of applied, and overall VA transport in runoff was predominately in the dissolved phase (90% for SMZ and 99% for LIN). Among vegetation treatments, only tall fescue significantly reduced loads of SMZ and LIN compared with the control, with load reductions of ∼30% for both VAs. Estimated field-scale reductions in VA loads showed that source-to-buffer area ratios (SBARs) of 10:1 to 20:1 reduced VA loads by only 7 to 16%. Overall, the grass VBS tested here were less effective at reducing SMZ and LIN loads in surface runoff than has been previously demonstrated for sediment, nutrients, and herbicides.

Journal of Environmental Quality

Microbial source tracking and land use associations for antibiotic resistance genes in private wells influenced by human and livestock fecal sources

Antimicrobial resistance is a growing public health problem that requires an integrated approach among human, agricultural, and environmental sectors. However, few studies address all three components simultaneously. We investigated the occurrence of five antibiotic resistance genes (ARGs) and the class 1 integron gene ( intI1 ) in private wells drawing water from a vulnerable aquifer influenced by residential septic systems and land-applied dairy manure. Samples ( n = 138) were collected across four seasons from a randomized sample of private wells in Kewaunee County, Wisconsin. Measurements of ARGs and intI1 were related to microbial source tracking (MST) markers specific to human and bovine feces; they were also related to 54 risk factors for contamination representing land use, rainfall, hydrogeology, and well construction. ARGs and intI1 occurred in 5–40% of samples depending on target. Detection frequencies for ARGs and intI1 were lowest in the absence of human and bovine MST markers (1-30%), highest when co-occurring with human and bovine markers together (11-78%), and intermediate when co-occurring with just one type of MST marker (4-46%). Gene targets were associated with septic system density more often than agricultural land, potentially because of the variable presence of manure on the landscape. Determining ARG prevalence in a rural setting with mixed land use allowed an assessment of the relative contribution of human and bovine fecal sources. Because fecal sources co-occurred with ARGs at similar rates, interventions intended to reduce ARG occurrence may be most effective if both sources are considered.

Wisconsin

A universal method for the simultaneous determination of environmental pollutants in marine biological samples: Per- and polyfluoroalkyl substances and antibiotics as a case study

Conventional detection technologies for environmental contaminants have primarily focused on providing accurate qualitative and quantitative evaluations for single pollutant types, leading to increased costs and an inability to satisfy the growing demand for detecting a broader spectrum of pollutants. Here, we introduced a novel analytical method to simultaneously measure the concentration levels of diverse environmental pollutants, characterized by their distinct properties, across complex biological samples. Per- and polyfluoroalkyl substances (PFAS) and antibiotics were used as a case study due to their frequency of detection in the environment and known impacts Our method harnesses the salting-out effect of sodium chloride on proteins within the muscle tissues of 178 marine species, which significantly reduces the addition of extraneous substances, mitigates matrix interference, and avoids reliance on solid-phase extraction or dispersive extraction agents. The method provides a simultaneous pretreatment for the detection of several compounds, with detection limits from 0.002 to 0.41 ng/g dry weight, which are substantially lower than conventional methods. Overall, this method streamlines efficiency, decreases costs, lessens matrix effects, and sets a solid groundwork for future applications in the concurrent detection of a broader spectrum of environmentally pertinent pollutants with varied characteristics.

ACS ES&T Water

Quantitative microbial risk assessment for ingestion of antibiotic resistance genes from private wells contaminated by human and livestock fecal sources

We used quantitative microbial risk assessment to estimate ingestion risk for intI1 , erm (B), sul1 , tet (A), tet (W), and tet (X) in private wells contaminated by human and/or livestock feces. Genes were quantified with five human-specific and six bovine-specific microbial source-tracking (MST) markers in 138 well-water samples from a rural Wisconsin county. Daily ingestion risk (probability of swallowing ≥1 gene) was based on daily water consumption and a Poisson exposure model. Calculations were stratified by MST source and soil depth over the aquifer where wells were drilled. Relative ingestion risk was estimated using wells with no MST detections and >6.1 m soil depth as a referent category. Daily ingestion risk varied from 0 to 8.8 × 10 −1 by gene and fecal source (i.e., human or bovine). The estimated number of residents ingesting target genes from private wells varied from 910 ( tet (A)) to 1,500 ( intI1 and tet (X)) per day out of 12,000 total. Relative risk of tet (A) ingestion was significantly higher in wells with MST markers detected, including wells with ≤6.1 m soil depth contaminated by bovine markers (2.2 [90% CI: 1.1–4.7]), wells with >6.1 m soil depth contaminated by bovine markers (1.8 [1.002–3.9]), and wells with ≤6.1 m soil depth contaminated by bovine and human markers simultaneously (3.1 [1.7–6.5]). Antibiotic resistance genes (ARGs) were not necessarily present in viable microorganisms, and ingestion is not directly associated with infection. However, results illustrate relative contributions of human and livestock fecal sources to ARG exposure and highlight rural groundwater as a significant point of exposure.

Wisconsin

Identifying sources of antibiotic resistance genes in the environment using the microbial Find, Inform, and Test framework

Introduction: Antimicrobial resistance (AMR) is an increasing public health concern for humans, animals, and the environment. However, the contributions of spatially distributed sources of AMR in the environment are not well defined. Methods: To identify the sources of environmental AMR, the novel microbial Find, Inform, and Test (FIT) model was applied to a panel of five antibiotic resistance-associated genes (ARGs), namely, erm(B), tet(W), qnrA, sul1, and intI1, quantified from riverbed sediment and surface water from a mixed-use region. Results: A one standard deviation increase in the modeled contributions of elevated AMR from bovine sources or land-applied waste sources [land application of biosolids, sludge, and industrial wastewater (i.e., food processing) and domestic (i.e., municipal and septage)] was associated with 34–80% and 33–77% increases in the relative abundances of the ARGs in riverbed sediment and surface water, respectively. Sources influenced environmental AMR at overland distances of up to 13 km. Discussion: Our study corroborates previous evidence of offsite migration of microbial pollution from bovine sources and newly suggests offsite migration from land-applied waste. With FIT, we estimated the distance-based influence range overland and downstream around sources to model the impact these sources may have on AMR at unsampled sites. This modeling supports targeted monitoring of AMR from sources for future exposure and risk mitigation efforts.

Wisconsin

Reconnaissance data for glyphosate, other selected herbicides, their degradation products, and antibiotics in 51 streams in nine midwestern states, 2002

Since 1989, the U.S. Geological Survey has conducted periodic reconnaissance studies of streams in the Midwestern United States to determine the geographic and seasonal distribution of herbicide compounds. These studies have documented that large amounts of acetochlor, alachlor, atrazine, cyanazine, metolachlor, and their degradation products are flushed into streams during post-application runoff. Additional studies show that peak herbicide concentrations tend to occur during the first runoff after herbicide application and that herbicide flushes can occur during runoff for several weeks to months following application. Since the first stream study conducted in 1989, several significant changes in herbicide use have occurred. The most substantial change is the tripling in the use of glyphosate during the past 5 years. Over this same time period (1997-2001), usage of acetochlor and atrazine increased slightly, whereas alachlor, cyanazine, and metolachlor usage decreased. During 2002, 154 samples were collected from 51 streams in nine Midwestern States during three periods of runoff. This report provides a compilation of the analytical results of five laboratory methods. Results show that glyphosate was detected in 55 (36 percent) of the samples, and aminomethylphosphonic acid (a degradation product of glyphosate) was detected in 107 (69 percent) of the samples. Atrazine, the most frequently detected herbicide, was found in 93 percent of the samples, followed by metolachlor, found in 73 percent of the samples; metolachlor ethanesulfonic acid (ESA) and oxanilic acid (OXA) were the most frequently detected herbicide degradation products, both being found in more than 95 percent of the samples. The data presented here are valuable for comparison with results from the earlier reconnaissance studies.

Minnesota, Wisconsin, Iowa, Illinois, Indiana, Ohi