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2000 Progress Report: Immunoassay and Serum Markers
EPA Grant Number: R825433C042Subproject: this is subproject number 042 , established and managed by the Center Director under grant R825433
(EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
Center: EERC - Center for Ecological Health Research (Cal Davis)
Center Director: Rolston, Dennis E.
Title: Immunoassay and Serum Markers
Investigators: Hammock, Bruce
Institution: University of California - Davis
EPA Project Officer: Levinson, Barbara
Project Period: October 1, 1996 through September 30, 2000
Project Period Covered by this Report: October 1, 1999 through September 30, 2000
RFA: Exploratory Environmental Research Centers (1992)
Research Category: Center for Ecological Health Research , Targeted Research
Description:
Objective:To develop rapid, inexpensive, and accurate analytical methods to determine the presence and effects of toxic chemicals in animals and the environment using immunoassays as indicators of exposure and serum esterases and lipid oxides as biomarkers of effect in animals.
Progress Summary:Immunoassays. The development and implementation of immunoassays for monitoring environmental compounds that can contribute to stress in varying components of the ecosystem has been the major focus. Assays for pyrethroid insecticides esfenvalerate and permethrin are being used in runoff studies along with GC analysis to validate the immunoassay methods for future studies. Although esfenvalerate and permethrin were known targets in this runoff study, other pyrethroids are also of interest. Assays that detect the class of pyrethroids, either as members of the class that yield phenoxybenzoic acid as a breakdown product, or as type I or type II pyrethroids have been also been developed for use when the identity of the pyrethroid may not be known, or a mixture may be involved. Assays for dioxins (especially TCDD) are being validated for use in a variety of matrices including soil, sediment, eggs, fish, milk and water.
We continue to work toward improvements in immunoassay as an analytical technique. One such improvement would be to use labels that fluoresce at wavelengths in the near infrared. Detection of such labels would be away from wavelengths where naturally fluorescing materials occur, thus reducing background interference. Two demonstration projects include an immunoassay for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) that used the phosphorescent label platinum coproporphyrin. The second project uses the glycine conjugate of phenoxybenzyl alcohol (a putative human metabolite of some pyrethroid insecticides) and a quenching fluoroimmunoassay. These assays are one-step compared to several for the traditional immunoassay. One of the most exciting developments has been the adaptation of ELISAs to compact disk (CD) surfaces allowing simple sample handling and high throughput. Currently the technology is in the patent process and the scientist involved in this work has moved to a company that is focusing on this technology.
Serum and Urine Markers. The release of enzymes from hepatic tissues into the serum can be an indication of multiple stresses in vertebrate species. This work is being conducted in collaboration with Dr. Barry Wilson. In another case we are examining lipid diols and epoxides as indicators of oxidative stress. We have found in preliminary experiments that not only are lipid epoxides indicators of stress, but that several of these materials in both the linoleate series and arachidonic acid series are very toxic. They thus are biomarkers both of exposure and of effect. We are investigating the idea that environmental stress can lead directly to lipid oxides (ozone and some nitrogen oxides will oxidize lipids directly), but we also are working on the hypothesis that foreign materials and possibly general stress can induce enzymes leading to the formation of these biomarkers. We will also test the hypothesis that some of the lipid oxidation products have a direct deleterious effect on animals. Although animals are our initial target we will work to extend these studies to plants in support of Dr. Fan. For instance we have cloned one of the key genes involved in lipid oxidation from two plant species and have prepared a knock out potato plant as a model species for study. We have developed a GC method for analysis of these oxylipins. These lipids are also excreted in the urine and an immunoassay is under development.
Other. Recombinant viral insecticides, as an alternative to chemical pesticides, can reduce environmental stress to organisms by reducing the exposure of these organisms to toxic environmental chemicals. Thus we are expressing recombinant peptides designed to disrupt the development of pest insects. It appears that these viruses can reduce dramatically the environmental impact associated with pest control. We are examining in turn possible deleterious effects of this technology on the ecosystem.
Future Activities:The projects listed above will be continued with several emphases. In the immunoassay area we will continue to respond to Center investigators needs for immunoassays. For example we are working on the development of assays to octyl and nonyl polyethoxylates and the breakdown products octyl- and nonyl phenol in collaboration with Dr. Mike Fry. Assays for pyrethroids and their breakdown products will be used to monitor pyrethroids during the upcoming dormant spray season. Recent studies in the Bay-Delta indicate that dioxins, particularly TCDD, may pose some risk. Our TCDD ELISA is sensitive to low ppt and can be used in support of several projects. In addition we are working on advancing the technology to make immunoassays more field portable, cheaper and more sensitive. Approaches include moving ELISA toward near IR based biosensors based on microfluidics to increase signal to noise. Another use of near IR labels is to adapt ELISAs toward more high throughput systems currently used in microarray or combinatorial chemistry techniques. We are also exploring novel mathematical approaches for data analysis that will take advantage of compound and class-selective assay responses to determine the identity and concentration of analyte. Finally, development of recombinant reporter systems and antibodies is also of interest.
The esterase work with Wilson is being continued. The lipid oxides and their metabolites as biomarkers of stress work described above will be emphasized.
Lastly we will continue to stress the use of accelerator mass spectrometry (AMS). Since there are only three of these instruments in the U.S., we are in a unique position to carry out this work. One can consider an AMS as a liquid scintillation counter with a 100,000 fold increase in sensitivity for certain ions. In these studies 14C is used as a tracer at levels that are far below those detectable with any sensor for radioactivity. We have found this technique to be very powerful in model studies with the herbicide atrazine. We have just finished a human metabolism study of the herbicide atrazine. This is the first pharmacokinetic study done in humans at environmentally relevant doses. The technology facilitates the use of C as a tracer in environmental studies at levels that are below regulatory limits for radioactivity. It is a technology which we hope will we widely applicable to ecological studies.
Supplemental Keywords:Ecosystem Protection/Environmental Exposure & Risk, Water, INTERNATIONAL COOPERATION, Geographic Area, Scientific Discipline, RFA, ECOSYSTEMS, Ecosystem/Assessment/Indicators, Water & Watershed, exploratory research environmental biology, Restoration, Aquatic Ecosystem Restoration, Aquatic Ecosystems & Estuarine Research, Terrestrial Ecosystems, Ecological Monitoring, Aquatic Ecosystem, Ecological Indicators, Biochemistry, Ecological Effects - Human Health, Watersheds, Environmental Chemistry, Ecological Effects - Environmental Exposure & Risk, Ecosystem Protection, Monitoring/Modeling, Ecology and Ecosystems, Environmental Monitoring, State, biomarkers, runoff, water quality, California (CA), wildlife habitat, watershed modeling, watershed land use, watershed, watershed management, watershed restoration, immunoassay, Clear Lake, ecological risk, ecology assessment models, ecosystem health, Clear Lake , environmental stress indicators, anthropogenic effects, fish habitat, land use, wetlands, accelerator mass spectrometry, watershed development, aquatic habitat, agricultural watershed, aquatic ecosystems, environmental stress, pesticides, lake ecosysyems, watershed sustainablility, ecological assessment, hydrology, water management options, ecosystem stress, serum markers, ecosystem response, ecosystem monitoring, nutrient dynamics
Progress and Final Reports:
1999 Progress Report
Original Abstract
Final Report
Main Center Abstract and Reports:
R825433 EERC - Center for Ecological Health Research (Cal Davis)
Subprojects under this Center:
(EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
R825433C001 Potential for Long-Term Degradation of Wetland Water Quality Due to Natural Discharge of Polluted Groundwater
R825433C002 Sacramento River Watershed
R825433C003 Endocrine Disruption in Fish and Birds
R825433C004 Biomarkers of Exposure and Deleterious Effect: A Laboratory and Field Investigation
R825433C005 Fish Developmental Toxicity/Recruitment
R825433C006 Resolving Multiple Stressors by Biochemical Indicator Patterns and their Linkages to Adverse Effects on Benthic Invertebrate Patterns
R825433C007 Environmental Chemistry of Bioavailability in Sediments and Water Column
R825433C008 Reproduction of Birds and mammals in a terrestrial-aquatic interface
R825433C009 Modeling Ecosystems Under Combined Stress
R825433C010 Mercury Uptake by Fish
R825433C011 Clear Lake Watershed
R825433C012 The Role of Fishes as Transporters of Mercury
R825433C013 Wetlands Restoration
R825433C014 Wildlife Bioaccumulation and Effects
R825433C015 Microbiology of Mercury Methylation in Sediments
R825433C016 Hg and Fe Biogeochemistry
R825433C017 Water Motions and Material Transport
R825433C018 Economic Impacts of Multiple Stresses
R825433C019 The History of Anthropogenic Effects
R825433C020 Wetland Restoration
R825433C021 Sierra Nevada Watershed Project
R825433C022 Regional Transport of Air Pollutants and Exposure of Sierra Nevada Forests to Ozone
R825433C023 Biomarkers of Ozone Damage to Sierra Nevada Vegetation
R825433C024 Effects of Air Pollution on Water Quality: Emission of MTBE and Other Pollutants From Motorized Watercraft
R825433C025 Regional Movement of Toxics
R825433C026 Effect of Photochemical Reactions in Fog Drops and Aerosol Particles on the Fate of Atmospheric Chemicals in the Central Valley
R825433C027 Source Load Modeling for Sediment in Mountainous Watersheds
R825433C028 Stress of Increased Sediment Loading on Lake and Stream Function
R825433C029 Watershed Response to Natural and Anthropogenic Stress: Lake Tahoe Nutrient Budget
R825433C030 Mercury Distribution and Cycling in Sierra Nevada Waterbodies
R825433C031 Pre-contact Forest Structure
R825433C032 Identification and distribution of pest complexes in relation to late seral/old growth forest structure in the Lake Tahoe watershed
R825433C033 Subalpine Marsh Plant Communities as Early Indicators of Ecosystem Stress
R825433C034 Regional Hydrogeology and Contaminant Transport in a Sierra Nevada Ecosystem
R825433C035 Border Rivers Watershed
R825433C036 Toxicity Studies
R825433C037 Watershed Assessment
R825433C038 Microbiological Processes in Sediments
R825433C039 Analytical and Biomarkers Core
R825433C040 Organic Analysis
R825433C041 Inorganic Analysis
R825433C042 Immunoassay and Serum Markers
R825433C043 Sensitive Biomarkers to Detect Biochemical Changes Indicating Multiple Stresses Including Chemically Induced Stresses
R825433C044 Molecular, Cellular and Animal Biomarkers of Exposure and Effect
R825433C045 Microbial Community Assays
R825433C046 Cumulative and Integrative Biochemical Indicators
R825433C047 Mercury and Iron Biogeochemistry
R825433C048 Transport and Fate Core
R825433C049 Role of Hydrogeologic Processes in Alpine Ecosystem Health
R825433C050 Regional Hydrologic Modeling With Emphasis on Watershed-Scale Environmental Stresses
R825433C051 Development of Pollutant Fate and Transport Models for Use in Terrestrial Ecosystem Exposure Assessment
R825433C052 Pesticide Transport in Subsurface and Surface Water Systems
R825433C053 Currents in Clear Lake
R825433C054 Data Integration and Decision Support Core
R825433C055 Spatial Patterns and Biodiversity
R825433C056 Modeling Transport in Aquatic Systems
R825433C057 Spatial and Temporal Trends in Water Quality
R825433C058 Time Series Analysis and Modeling Ecological Risk
R825433C059 WWW/Outreach
R825433C060 Economic Effects of Multiple Stresses
R825433C061 Effects of Nutrients on Algal Growth
R825433C062 Nutrient Loading
R825433C063 Subalpine Wetlands as Early Indicators of Ecosystem Stress
R825433C064 Chlorinated Hydrocarbons
R825433C065 Sierra Ozone Studies
R825433C066 Assessment of Multiple Stresses on Soil Microbial Communities
R825433C067 Terrestrial - Agriculture
R825433C069 Molecular Epidemiology Core
R825433C070 Serum Markers of Environmental Stress
R825433C071 Development of Sensitive Biomarkers Based on Chemically Induced Changes in Expressions of Oncogenes
R825433C072 Molecular Monitoring of Microbial Populations
R825433C073 Aquatic - Rivers and Estuaries
R825433C074 Border Rivers - Toxicity Studies