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1999 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, 1998 through September 30, 1999
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 as biomarkers of effect in animals.
Progress Summary:Our major effort has been in the development and implementation of immunoassays for monitoring environmental compounds that can contribute to stress in varying components of the ecosystem. When addressing questions of multiple stressors, large numbers of samples for specific compounds are generated. There are numerous analytical methods available, however immunoassays are well-known to be rapid, inexpensive for large sample loads and to utilize simple equipment. In addition, assays that detect classes of compounds are needed so that sets of molecules that may cause stress can be analyzed together. Thus, we have developed assays that are highly specific such as for the insecticide esfenvalerate. In the case of esfenvalerate, the assays are also stereoselective. We have pioneered the technique of designing class specific assays which will detect many or all members of a class of compounds such as mercapturates or glucuronides. We can use compound- and class-selective assays to distinguish the subclasses of compounds using mathematical approaches based on immunoassay arrays. The triazine herbicides illustrate this approach where we have assays for this major class of compounds and other assays which distinguish individual members of the group.
In addition, we are striving to improve over all analytical technology in the immunoassay area. The development of novel immunoassay formats, the evaluation of near infrared technology to develop immunoassays which can be run faster, with greater sensitivity and with less expensive equipment and use of fluorescent molecules as labels are three examples.
As a lower priority we are addressing two other topics. In one case we are looking at the release of enzymes from hepatic tissues into the serum as an indication of multiple stresses in vertebrate species. This work is in collaboration with Dr. Barry Wilson. In another case we are looking at lipid diols and epoxides as indicators of oxidative stress. This involves analysis of these oxylipins in urine and other body fluids as well as an investigation of the toxicity of these lipids to an individual organism.
The final topic relates to the development of recombinant viral insecticides. The concept of this work is that we 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 attempt to be responsive to Center investigators needs for additional assays. Pyrethroid assays recently developed at the request of Hinton (A.3) will be used to monitor pyrethroids in laboratory and field exposure studies. These compounds are very toxic to fish and other aquatic organisms. In addition we are developing assays to metabolites and environmental breakdown products for use in these monitoring studies. 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 to increase signal to noise assays, novel mathematical approaches for data analysis, and development of recombinant reporter systems and antibodies. One of the most exciting developments in this area has been the adaptation of ELISAs to compact disk (CD) surfaces allowing simple sample handling and high through put.
The esterase work with Wilson is being continued. Lipid oxides and their metabolites as biomarkers of stress will be emphasized. We have not only found in preliminary experiments that lipid epoxides are indicators of stress, we have found 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 (oxone 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 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 are developing both GLC/MS techniques and ELISA's for these biological indicators.
Finally 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. It is a technology which we hope will we widely applicable to ecological studies. 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.
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:
Original Abstract
2000 Progress Report
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