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Biosurveillance, Agricultural and Environmental Security: A Coordinated, Innovative Initiative

Schultz, Terry
University of Tennessee
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The objectives of this project are several. They include the development of: bacteriophage-based bioluminescent reporting systems to identify the presence of pathogenic agents in a stand off format and a proof-of-concept for algal- and higher plant-based sensors to detect chemical or biological agents in the environment. These objectives include adapting a self-catalytic bioluminescence system for transgenic reporting-based system using constitutive and inducible promoters. The objectives also include proof of concepts for using mammalian cell cultures and commercial arrays and bacterial-based bioluminescent biosensor in toxicogenomic evaluations. The former will focus on to provide toxicogenomic data for apoptotic events associated with immunotoxicty. Both genomics projects will include structure-activity analysis of gene expression data as a means of determining mechanisms of toxic action.
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NON-TECHNICAL SUMMARY: As concern over bio- and chemical-terrorism become more focused the need for a biological and chemical surveillance web based on unmanned sensors, remote sensing, and biotechnology and useful in detecting and tracking selected biological and chemical agents has become more apparent. Currently no demonstrated technologies exist on which to base such a web. This project examines several systems including bacterial, plant and animal cells, which use gene expression reporter technology in an attempt to form the base for a biological and chemical surveillance web.

APPROACH: The approach is to use molecular and other modern techniques to develop advanced bio-based sensors and computational toxicology to either sense or predict epidemiological patterns in real world scenarios. Bio-based sensors will focus on using Lux-based and green-fluorescent-protein-based technology to develop novel bacteriophage-based and plant-based sensors to detect biological agents (e.g., anthrax) or chemical agents (e.g., munitions). The bacteriophage sensors are being designed to take advantage of the specificity of phages to a particular type of bacteria so one can sense, identity, and communicate the presence of a particular pathological agent in the environment. The goal here is to develop stand off systems for tracking the presence and movement of biological and chemical agents. The initial work is with the foodborne pathogen E.coli O157:H7. The plant sensors are being engineered as sentinels or environmental monitors for the presence of chemical warfare products. Here the expression of a report gene is to be used to note the presence of a particular compound or class of compounds. In addition, genomic technology is to be used to explore the use of gene expression patterns to indicate exposure and form a basis for predictivity of effects. This aspect of the project focuses on using rapid and inexpensive in vitro systems to develop gene expression profiles to selected chemical agents in particular polycyclic aromatic hydrocarbons. In one aspect of the project human cell lines and commercial gene arrays are being explored as a means of detecting exposure to chemicals causing programmed cell death and immunotoxicity. In another aspect of the project a bacterial-based gene fusion system is being used to develop a data matrix as a means of comparing gene expression patterns and toxicity.

Funding Source
Nat'l. Inst. of Food and Agriculture
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Prevention and Control
Escherichia coli
Bacterial Pathogens