Validation of an extraction method for Cry1Ab protein from soil.

Corn expressing insecticidal proteins derived from Bacillus thuringiensis (Bt corn) has increased in usage in the United States from 8% of total corn acreage in 1996 to 67% in 2012. Because of this increase, it is important to be able to monitor the fate and transport of the insecticidal Bt proteins to evaluate environmental exposure and effects. Accurate and validated methods are needed to quantify these proteins in environmental matrices. A method to extract Bt Cry1Ab proteins from 3 soil types using a 10× phosphate-buffered saline with Tween buffer and a commercially available enzyme-linked immunosorbent assay (ELISA) was validated through a series of 6 tests. The validation process for Cry1Ab extractions in soil has not yet been reported in the scientific literature. The extraction buffer and each soil matrix were tested and validated for the ELISA. Extraction efficiencies were 41%, 74%, and 89% for the 3 soil types and were significantly correlated with the organic matter content of the soil. Despite low recoveries, consistent results with low coefficients of variation allowed for accurate measurements. Through validating this method with 3 different soils, a sensitive, specific, precise, and accurate quantification of Bt Cry1Ab was developed. The validation process can be expanded and implemented in other environmental matrices, adding consistency to data across a wide range of samples.

Determining modifications to bifenthrin toxicity and sediment binding affinity from varying potassium chloride concentrations in overlying water.

Bifenthrin, a current-use pyrethroid insecticide, has been repeatedly identified as a major contributor to toxicity in urban and residential stream sediment. Within an urban stream multiple stressors exist. However, other than pesticides, the influence of secondary stressors on bifenthrin toxicity has not been studied. The goal of this project was to study how dissolved ions, based on the model salt KCl, influence bifenthrin toxicity. The presence of these dissolved ions could influence bifenthrin toxicity either through joint action as a secondary toxicant or through changing the partitioning or bioavailability of bifenthrin between the sediment matrix and overlying water or pore water. The first objective was to determine if mixtures of bifenthrin and KCl, a commonly utilized reference toxicant, display additive toxicity to the benthic invertebrates Hyalella azteca and Chironomus dilutus using concentration addition and independent action mathematical models. The second objective of the present study was to examine how KCl dissolved in the overlying water influences partitioning and bioavailability of a pyrethroid (bifenthrin). Joint toxicity of bifenthrin and KCl was less than predicted by both concentration addition and independent action models. However, both models predicted the joint toxicity within a factor of two. Partitioning of bifenthrin was not significantly influenced by KCl concentrations based on K(oc) determinations and desorption to Tenax beads. This indicates that the fate and bioavailability of bifenthrin are not likely different in aquatic environments with varying dissolved ion concentrations. Therefore, the toxicological interaction that results in the antagonistic joint action between bifenthrin and KCl is likely due to the physiological effects of exposure to hypertonic solutions of KCl rather than alterations to bifenthrin bioavailability.

Mercury concentrations in Quagga Mussels, Dreissena bugensis, from Lakes Mead, Mohave and Havasu.

The recent invasion of the Dressenid species, the quagga mussel, Dreissena bugensis, into Lakes Mead, Mohave and Havasu has raised questions about their ability to alter contaminant cycling. Mussels were collected from 25 locations in the three lakes. The overall average was 0.036 +/- 0.016 microg g(-1) Hg dry wt. The range of the three lakes was from 0.014-0.093 microg g(-1) Hg dry wt. There were no significant differences in mercury concentrations among the three lakes (F = 0.07; p = 0.794). From this baseline data of contaminants in quagga mussels from the lower Colorado River, this species may be used to biomonitor lake health.