Claudia Sheedy

Development of competitive ELISAs for 17β-estradiol and 17β-estradiol +estrone+estriol using rabbit polyclonal antibodies

Estrogens are a family of feminizing hormones that are excreted by vertebrates. It has been documented that their presence in surface waters, even in the ng/L range, can have detrimental impacts on fish reproduction. Two competitive enzyme-linked immunosorbent assays using rabbit polyclonal antibodies were developed: one for 17β-estradiol and a second one for 17β-estradiol (E2)+estrone (E1)+estriol (E3). Two different conjugates were synthesized using the Mixed-anhydride (for the 17β-estradiol ELISA) and the Mannich (for the E1 + E2 + E3 ELISA) reactions. The 17β-estradiol ELISA was highly specific with an IC50 of 243 ng/mL for 17β-estradiol. The E1 + E2 + E3 ELISA exhibited cross-reactivity with estrone (85%) and estriol (62%) with an IC50 of 18 ng/mL for 17β-estradiol. Cross-reactivity was tested against 13 chemically related compounds and both immunoassays showed significant cross-reactivity with two estradiol conjugates: β estradiol-17-valerate and β estradiol-3-benzoate (from 57 to 84 %) for which, to our knowledge, there are currently no commercially available ELISA. Characteristics (sensitivity, inter and intra assay variation, and cross-reactivity) of the E1 + E2 + E3 ELISA were further compared to those from a commercial Estriol ELISA. The commercial ELISA was more specific, sensitive and its inter-assay variation was less (9.5% compared to 10% for the E1 + E2 + E3 ELISA) but the E1 + E2 + E3 ELISA had less intra-assay variation (4% compared to 5% for the commercial ELISA). Finally, a solid-phase extraction method compatible with the E1 + E2 + E3 immunoassay demonstrated that this combined approach of extraction and immunoassay had good potential for determining estrogen concentrations in environmental samples such as surface water in urban and agricultural ecosystems.

A coupled stochastic/deterministic model to estimate the evolution of the risk of water contamination by pesticides across Canada

Periodic assessments of the risk of water contamination by pesticides help decision makers improve the sustainability of agricultural management practices. In Canada, when evaluating the risk of water contamination by pesticides, 2 main constraints arise. First, because the area of interest is large, a pesticide transport model with low computational running time is mandatory. Second, some relevant input data for simulations are not known, and most are known only at coarse scale. This study aims to develop a robust methodology to estimate the evolution of the risk of water contamination by pesticides across Canada. To circumvent the 2 aforementioned issues, we constructed a stochastic model and coupled it to the 1-dimensional pesticide fate model Pesticide Root Zone Model (PRZM). To account for input data uncertainty, the stochastic model uses a Monte Carlo approach to generate several pesticide application scenarios and to randomly select PRZM parameter values. One hundred different scenarios were simulated for each of over 2000 regions (Soil Landscapes of Canada [SLC] polygons) for the years 1981 and 2006. Overall, the results indicated that in those regions in which the risk increased from 1981 to 2006, the increase in risk was mainly attributable to the increased area treated by pesticides or an increase in the number of days with runoff. More specifically, this work identifies the areas at higher risk, where further analyses with finer-scale input data should be performed. The model is specific for Canadian data, but the framework could be adapted for other large countries.

Development of a real-time immuno-PCR assay for the quantification of 17β-estradiol in water.

A competitive real-time (RT) immuno-polymerase chain reaction (iPCR) (RT-iPCR) assay was developed for the sensitive quantification of 17β-estradiol in water. Using a universal iPCR method and polyclonal antibodies, 17β-estradiol was accurately quantified at concentrations ranging from 1 pg mL−1 to 10 µg mL−1. The RT-iPCR assays limit of detection was 0.7 pg mL−1. The RT-iPCR assay provided an 800-fold increase in sensitivity as well as an expanded working range compared with the corresponding enzyme-linked immunosorbent assay. Assay cross-reactivity to estrone and estriol, two structurally related estrogens, was below 8%. Water samples spiked with 17β-estradiol were analyzed by RT-iPCR to determine the assays potential as a rapid screen for the monitoring of manure-borne estrogens in the environment. The assay showed recoveries of 82, 102 and 103% for Milli-Q, tap, and irrigation water, respectively, without requiring sample extraction or concentration prior to analysis.

Solid beef cattle manure application impacts on soil properties and 17β-estradiol fate in a clay loam soil.

Livestock manure applied to agricultural land is one of the ways natural steroid estrogens enter soils. To examine the impact of long-term solid beef cattle (Bos Taurus) manure on soil properties and 17β-estradiol sorption and mineralization, this study utilized a soil that had received beef cattle manure over 35 years. The 17β-estradiol was strongly sorbed and sorption significantly increased (P < 0.05) with increasing soil organic carbon content (SOC) and with an increasing annual rate of beef cattle manure. The 17β-estradiol mineralization half-life was significantly negatively correlated, and the total amount of 17β-estradiol mineralized at 90 days (MAX) was significantly positively correlated with 17β-estradiol sorption. The long-term rate of manure application had no significant effect on MAX, but the addition of fresh beef cattle manure in the laboratory resulted in significantly (P < 0.05) smaller MAX values. None of the treatments showed MAX values exceeding one-third of the 17β-estradiol applied.

Integrated assessment of climate change impact on surface runoff contamination by pesticides

Pesticide transport by surface runoff depends on climate, agricultural practices, topography, soil characteristics, crop type, and pest phenology. To accurately assess the impact of climate change, these factors must be accounted for in a single framework by integrating their interaction and uncertainty. This article presents the development and application of a framework to assess the impact of climate change on pesticide transport by surface runoff in southern Québec (Canada) for the 1981-2040 period. The crop enemies investigated were: weeds for corn (Zea mays); and for apple orchard (Malus pumila), 3 insect pests (codling moth [Cydia pomonella], plum curculio [Conotrachelus nenuphar], and apple maggot [Rhagoletis pomonella]), 2 diseases (apple scab [Venturia inaequalis], and fire blight [Erwinia amylovora]). A total of 23 climate simulations, 19 sites, and 11 active ingredients were considered. The relationship between climate and phenology was accounted for by bioclimatic models of the Computer Centre for Agricultural Pest Forecasting (CIPRA) software. Exported loads of pesticides were evaluated at the edge-of-field scale using the Pesticide Root Zone Model (PRZM), simulating both hydrology and chemical transport. A stochastic model was developed to account for PRZM parameter uncertainty. Results of this study indicate that for the 2011-2040 period, application dates would be advanced from 3 to 7 days on average with respect to the 1981-2010 period. However, the impact of climate change on maximum daily rainfall during the application window is not statistically significant, mainly due to the high variability of extreme rainfall events. Hence, for the studied sites and crop enemies considered, climate change impact on pesticide transported in surface runoff is not statistically significant throughout the 2011-2040 period. Integr Environ Assess Managem 2016;12:559-571.

Auxin Herbicides and Pesticide Mixtures in Groundwater of a Canadian Prairie Province

Groundwater samples were collected from piezometers and water table wells in both dryland and irrigated agricultural regions of Alberta, Canada, to examine the occurrence of pesticide mixtures. Fourteen current-use pesticides and two historical compounds were detected over a 3-yr sampling period. Pesticide mixtures were detected in ∼3% of the groundwater samples, and the frequency of detection increased from spring (1.5%) to summer (3.8%) and fall (4.8%). Pesticide mixtures always consisted of at least one of two auxin herbicides: 2,4-dichlorophenoxyacetic acid (2,4-D) or 2-methyl-4-chlorophenoxyacetic acid (MCPA). 19% of all samples contained a single pesticide, with auxin herbicides 2,4-D (7.3%), MCPA (4.4%), and clopyralid (3.9%) being most prevalent. We detected 2,4-D predominantly in the fall (72% of 2,4-D detections) and less in spring and summer (28%). We detected MCPA mostly in summer (85% of MCPA detections) and less in spring and fall (15%). Clopyralid was more evenly detected across spring (30%), summer (25%), and fall (45%). Since the auxin herbicides above are typically applied in summer, results suggest that each herbicide may have different mobility and persistence characteristics in prairie soils. Guidelines for Canadian Drinking Water Quality have been set for a range of individual pesticides, but not for pesticide mixtures. If Canada is to establish such guidelines, this study demonstrates that auxin herbicides should be prioritized. In addition, only 7 of the 16 compounds detected in this study have established maximum acceptable concentrations (MACs), excluding clopyralid, which was detected in all three sampling years.