Francis X. M. Casey

Development of an ultra-high-pressure liquid chromatography-tandem mass spectrometry multi-residue sulfonamide method and its application to water, manure slurry, and soils from swine rearing facilities.

An analytical method was developed using ultra-high-pressure liquid chromatography-triple quadrupole-tandem mass spectrometry (UHPLC-TQ-MS/MS) to simultaneously analyze 14 sulfonamides (SA) in 6 min. Despite the rapidity of the assay the system was properly re-equilibrated in this time. No carryover was observed even after high analyte concentrations. The instrumental detection limit based on signal-to-noise ratio (S/N)>3, was below 1 pg/microL (5 pg on column) for all SAs except sulfachloropyridazine. Surface water, ground water, soil, and slurry manure contained in storage ponds in and around swine [Sus scrofa domesticus] rearing facilities were analyzed. Sample cleanup for ground water and surface water included using solid phase extraction (SPE) using Oasis hydrophilic-lipophilic balance (HLB) cartridges. The soil and slurry manure required tandem strong anion exchange (SAX) and HLB solid phase extraction cartridges for sample cleanup. With few exceptions, the recoveries ranged from 60 to 100% for all matrices. The minimum detectable levels were below 2.0 ng/L for water, 30 ng/L for slurry manure, and 45 ng/kg for soil except for sulfachloropyridazine. The coefficient of variation (CV) was within 20% for most of the compounds analyzed. Using this method, sulfamethazine concentrations of 2250-5060 ng/L, sulfamethoxazole concentrations of 108-1.47 x 10(6)ng/L, and sulfathiazole concentrations of 785-1700 ng/L were found in the slurry manure. Sulfadimethoxine (2.0-32 ng/L), sulfamethazine (2.0-5.1 ng/L), and sulfamethoxazole (20.5-43.0 ng/L) were found in surface water and ground water. In top soil (0-15 cm), sulfamethazine ranged 34.5-663 ng/kg dry weight in those locations that received slurry manure as a nutrient; no SAs were found in the soil depths between 46 and 61 cm. The speed makes the method practical for medium to high throughput applications. The sensitivity and positive analyte identification make the method suitable for the demanding requirements for real world applications.

Atrazine removal in agricultural infiltrate by bioaugmented polyvinyl alcohol immobilized and free Agrobacterium radiobacter J14a: A sand column study

Bench-scale sand column breakthrough experiments were conducted to examine atrazine removal in agricultural infiltrate by Agrobacterium radiobacter J14a (J14a) immobilized in phosphorylated-polyvinyl alcohol compared to free J14a cells. The effects of cell loading and infiltration rate on atrazine degradation and the loss of J14a were investigated. Four sets of experiments, (i) tracers, (ii) immobilized dead cells, (iii) immobilized cells, and (iv) free cells, were performed. The atrazine biodegradation at the cell loadings of 300, 600, and 900 mg dry cells L(-1) and the infiltration rates of 1, 3, and 6 cm d(-1) were tested for 5 column pore volumes (PV). The atrazine breakthrough results indicated that the immobilized dead cells significantly retarded atrazine transport. The atrazine removal efficiencies at the infiltration rates of 1, 3, and 6 cm d(-1) were 100%, 80-97%, and 50-70%, respectively. Atrazine degradation capacity for the immobilized cells was not significantly different from the free cells. Both infiltration rate and cell loading significantly affected atrazine removal for both cell systems. The bacterial loss from the immobilized cell system was 10-100 times less than that from the free cell system. For long-term tests at 50 PV, the immobilized cell system provided consistent atrazine removal efficiency while the atrazine removal by the free cells declined gradually because of the cell loss.

Occurrence and pathways of manure-borne 17β-estradiol in vadose zone water

The hormone 17beta-estradiol (E2) can cause endocrine disruption in sensitive species at part per trillion concentrations. The persistence and transport pathways of manure-borne E2 in agricultural soils were determined by comparing its occurrence with the transfer of water and the transport of non-sorbing fluorobenzoic acid (FBA) tracers. This comparison was done using capillary wick lysimeters installed 0.61m beneath three corn (Zea mays L.) plots that receive swine (Sus scrofa domesticus) manure from various sources. An additional control plot was included that received no manure. Soil water transfer was modeled to compare actual versus predicted percolation. On average, lysimeters collected 61% of the expected percolation and 8% of the FBA. There were frequent E2 detections, where there were an average of 8 detections for the 11 sample events. The average detection was 21ngL(-1) and its range was 1-245ngL(-1). 17beta-Estradiol was detected before manure was applied and also in the control plot lysimeters. Furthermore, the average mass recovery of E2 in all the lysimeters was >50%, which was greater than the FBA tracer recovery. Results indicated that tracer was transported with precipitated water infiltrating into the soil surface and percolating down through the soil profile. There was substantial evidence for antecedent E2, which was persistent and mobile. The persistence and mobility of the E2 may result from its associations with colloids, such as dissolved organic matter. Furthermore, this antecedent E2 appeared to overwhelm any observable effect of manure management on E2 fate and transport.

An on-farm survey of spatial and temporal stratifications of 17β-estradiol concentrations

17β-estradiol (E2) is a natural estrogenic hormone found in animal manure and urine, which may cause endocrine disruption in sensitive organisms. 17β-Estradiol has been widely detected in the environment, and animal agriculture may be an important source. The objectives of this study were to investigate the potential sources and/or spatial and temporal characteristics contributing to detections of E2 at the farm-scale. Soil cores, segmented into 0.15m increments, were taken down to the water table from four locations (three potential E2 sources and one control) at or around a swine (Sus scrofa domesticus) farm on five different dates between 2006 and 2007. Estradiol was quantified in the soil-water extracts using liquid chromatography-with tandem mass spectrometry (LC/MS/MS) analysis. Estradiol detections were widespread and found in 128 out of 345 extractions (37%). Concentrations ranged from 0 to 1910 ng L⁻¹ (porewater equivalents). A location that received field application of manure had significantly lower E2 concentrations compared to other locations. Also, the spring 2007 E2 concentrations were significantly higher than all other sample dates, perhaps related to climatic and hydrological events. Results suggested E2 was not directly related to manure sources, but was widespread in this environment. Where E2 was detected, highest concentrations favored the upper profile, while the greatest frequency of detections was in the lower profile and near the water table. Detections of E2 were associated with high organic mater contents in the upper profile and high sand contents in the lower profile. The study suggests that E2 is widespread in some soil environments, raising questions as to the source and mechanisms that facilitate its transport and mobility.

Effects of field-manure applications on stratified 17β-estradiol concentrations

The occurrence of the manure-borne estrogen, 17β-estradiol (E2), was investigated in laboratory and field soils. In the laboratory, E2 was applied to soil to simulate concentrations found in swine (Sus scrofa domestica) manure (5000ngL(-1)). The aqueous-extracted E2 dissipated in the soil by 98% within 1h and was not significantly different from background concentrations (18ng L(-1)) for the duration of the experiment (64h). In the field study, soil cores were taken before and several dates after swine manure application. Equivalent porewater concentrations of water-extractable E2 were determined in 0.15-m increments down to the water table (0.70-2.00m deep). The average frequency of detection for 168 samples was 38% (average=40ng L(-1) porewater equivalents). Eleven days after manure application there was no significant effect on E2 detection frequency or concentration. However, E2 concentrations significantly increased by 6 months after manure application, and appeared to be related to precipitation. Concentrations then returned to original levels by 17 months after manure application. Manure did not have an immediate effect on E2 occurrence due to the capacity of the soil to rapidly sorb E2. However, it appears that soil may act as a long-term reservoir for E2 in the environment, which may be periodically released through desorption.

Sorption and degradation of 17β-estradiol-17-sulfate in sterilized soil-water systems.

To identify abiotic processes that govern the fate of a sulfate conjugated estrogen, 17β-estradiol-17-sulfate (E2-17S), soil batch experiments were conducted to investigate the dissipation, sorption, and degradation of radiolabeled E2-17S under sterilized conditions. The aqueous dissipation half-lives (DT50) for E2-17S ranged from 2.5 to 9.3h for the topsoil of high organic carbon (OC) content (1.29%), but E2-17S remained at ∼80% of applied dose in the low OC (0.26%) subsoil by 14 d. The non-linear sorption isotherms indicated limited sorption of E2-17S, and the concentration-dependent log KOC values were 2.20 and 2.45 for the topsoil and subsoil, respectively. Additionally, two types of hydroxyl E2-17S (OH-E2-17S and diOH-E2-17S) were found as major metabolites in the aqueous phase, which represented 9-25% and 6-7% of applied dose for the topsoil and subsoil at 14 d, respectively. Free estrogens, 17β-estradiol (E2) and estrone (E1), were detected from the sorbed phase of the soil-water systems.

Dissipation and transformation of 17β-estradiol-17-sulfate in soil–water systems

In the environment, estrogen conjugates can be precursors to the endocrine-disrupting free estrogens, 17β-estradiol (E2) and estrone (E1). Compared to other estrogen conjugates, 17β-estradiol-17-sulfate (E2-17S) is detected at relatively high concentrations and frequencies in animal manure and surface runoff from fields receiving manure. To elucidate the lifecycle of manure-borne estrogens and their conjugates in the environment, the fate of radiolabelled E2-17S in agricultural soils was investigated using laboratory batch studies with soils of different organic carbon (OC) content (1.29% for topsoil versus 0.26% for subsoil). E2-17S was found relatively persistent in the aqueous phase throughout the duration of the 14 d experiment. The aqueous E2-17S persisted longer in the subsoil (half-lives (DT₅₀)=64-173 h) than the topsoil (DT₅₀=4.9-26 h), and the aqueous persistence of E2-17S depended on its initial concentration. The major transformation pathway was hydroxylation, yielding mono- and di-hydroxy-E2-17S (OH-E2-17S and diOH-E2-17S). Free estrogens, E2 and E1, were only observed in the sorbed phase of the soil at low concentrations (∼1% of applied dose), which demonstrated that deconjugation and subsequent oxidation had occurred. Although deconjugation was not a major pathway, E2-17S could be a precursor of free estrogens in the environment.

Potential bioactivity and association of 17β-estradiol with the dissolved and colloidal fractions of manure and soil

The dissolved (DF) and colloidal fractions (CF) of soil and manure play an important role in the environmental fate and transport of steroidal estrogens. The first objective of this study was to quantify the association of 17β-estradiol (E2) with the DF and CF isolated from (i) liquid swine manure (LSM), (ii) a soil:water mixture (soil), and (iii) a LSM:soil:water mixture (Soil+LSM). The appropriate CF and DF size fractions of the Soil, Soil+LSM, and LSM media were obtained by first filtering through a 0.45 μm filter, which provided the combined DF and CF (DF/CF). The DF/CF from the three media was spiked with carbon-14 ([(14)C]) radiolabeled E2 ([(14)C]-E2), and then ultrafiltered to isolate the CF (<0.45 μm and >1 kDa) from the DF (<1 kDa). The average recoveries of the [(14)C] associated with the DF were 67%-72%, 67%-79%, and 76%-78% for the Soil, Soil+LSM and LSM, respectively. For the CF that was retained on the 1 kDa filter, organic carbon and [(14)C]-E2 were dislodged with subsequent water rinses the Soil+LSM and LSM, but not the Soil. The second objective was to evaluate whether the E2 associated with the various fractions of the different media could still bind the estrogen receptor using an E2 receptor (17β-ER) competitor assay, which allowed E2 equivalent concentrations to be determined. The estrogen receptor assay results indicated that E2 present in the DF of the Soil and Soil+LSM solutions could still bind the estrogen receptor. Results from this study indicated that E2 preferentially associated with the DF of soil and manure, which may enhance its dissolved advective transport in surface and subsurface water. Furthermore, this study indicated that E2 associated with DF solutions in the environment could potentially induce endocrine responses through its interactions with estrogen receptor.

Field-scale relationships among soil properties and shallow groundwater quality.

It is important to understand the link between land surface/soil properties and shallow groundwater quality. To that end, soil properties and near-water-table groundwater chemistry of a shallow, unconfined aquifer were measured on a 100-m grid on a 64-ha irrigated field in southeastern North Dakota. Soil properties and hydrochemistry were compared via multivariate analysis that included product-moment correlations and factor analysis/principal component analysis. Topographic low areas where the water table was in close proximity to the soil surface generally had higher apparent electrical conductivity (ECa ) and higher percent silt and clay than higher positions on the landscape. The majority of the groundwater was characterized by Ca- and Mg-HCO3 type water and was associated with topographic high areas with lower ECa and net groundwater recharge. Small topographic depressions were areas of higher ECa (net groundwater discharge) where salts that precipitated via evapotranspiration and evaporative discharge dissolved and leached to the groundwater during short-term depression-focused recharge events. At this site, groundwater quality and soil ECa were related to surface topography. High-resolution topography and EC(a) measurements are necessary to characterize the land surface/soil properties and surficial groundwater quality at the field-scale and to delineate areas where the shallow groundwater is most susceptible to contamination.

A feasibility study of immobilized and free mixed culture bioaugmentation for treating atrazine in infiltrate

A feasibility study of phosphorylated-polyvinyl alcohol immobilized and free mixed bacterial culture bioaugmentation for removing atrazine in agricultural infiltrate was conducted utilizing a sand column setup. The effects of bacterial cell loading and infiltration rate on atrazine degradation were investigated by short-term tests in which the amount of synthetic infiltrate fed through was five times of the void volume (five pore volumes) of the sand column. In addition, the loss of the inoculated atrazine-degrading cultures and the change of bacterial community were determined. Selected tests were continued for monitoring a long-term performance of the system (50 pore volumes of the sand column). The results indicated that the inoculated cells removed 42-80% of the atrazine. The infiltration rate and cell loading significantly affected the atrazine removal. In the short-term tests, the immobilized and free cells provided similar atrazine removal; however, leaching of the free cells was much greater than that of the immobilized cells. For the long-term performance, only the immobilized cells provided consistent atrazine removal efficiency throughout the test. Both immobilized and free cell systems exhibited a significant change in bacterial community structure during the atrazine degradation experiments. The infiltration rate was a significant factor for the change.