Bioluminescent reporter bacteria detect contaminants in soil samples

Burlage, Robert S.; Palumbo, Anthony V.; Heitzer, A.; Sayler, Gary S.

doi:10.1007/bf02941845

Cited by 44 publications

(27 citation statements)

References 6 publications

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“…For our model system, we chose a bacterial biosensor for the detection of naphthalene, which is relatively poorly water soluble and hence a good model for the behavior of a large class of hydrophobic compounds with moderate gas phase partition constants (Henry constant, 0.02) (25). Bacterial biosensors for naphthalene have been described and used before (3,7,13,14,20), but they have not been systematically tested for accurate measurement of naphthalene through the gas phase, although the principle has been applied previously (23). Here we show that bacterial biosensors can be optimally maintained in controlled chemostat cultivation, react very reliably to naphthalene exposure with a proportional bioluminescence signal, and can be applied in gas phase measurements to increase the bioavailable amount of naphthalene to the cells, thereby effectively lowering the detectable naphthalene concentration range.…”

mentioning

confidence: 99%

Measurement of Biologically Available Naphthalene in Gas and Aqueous Phases by Use of a Pseudomonas putida Biosensor

Werlen

Jaspers

Meer

2004

Appl Environ Microbiol

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Genetically constructed microbial biosensors for measuring organic pollutants are mostly applied in aqueous samples. Unfortunately, the detection limit of most biosensors is insufficient to detect pollutants at low but environmentally relevant concentrations. However, organic pollutants with low levels of water solubility often have significant gas-water partitioning coefficients, which in principle makes it possible to measure such compounds in the gas rather than the aqueous phase. Here we describe the first use of a microbial biosensor for measuring organic pollutants directly in the gas phase. For this purpose, we reconstructed a bioluminescent Pseudomonas putida naphthalene biosensor strain to carry the NAH7 plasmid and a chromosomally inserted gene fusion between the sal promoter and the luxAB genes. Specific calibration studies were performed with suspended and filter-immobilized biosensor cells, in aqueous solution and in the gas phase. Gas phase measurements with filter-immobilized biosensor cells in closed flasks, with a naphthalene-contaminated aqueous phase, showed that the biosensor cells can measure naphthalene effectively. The biosensor cells on the filter responded with increasing light output proportional to the naphthalene concentration added to the water phase, even though only a small proportion of the naphthalene was present in the gas phase. In fact, the biosensor cells could concentrate a larger proportion of naphthalene through the gas phase than in the aqueous suspension, probably due to faster transport of naphthalene to the cells in the gas phase. This led to a 10-fold lower detectable aqueous naphthalene concentration (50 nM instead of 0.5 M). Thus, the use of bacterial biosensors for measuring organic pollutants in the gas phase is a valid method for increasing the sensitivity of these valuable biological devices.Increasingly, genetically modified bacteria are being developed and tested for use as measuring devices for pollutants in the environment (reviewed recently in references 5, 8, and 19). Such bacterial biosensors either constitutively express a signal, such as bioluminescence, with any signal decrease related to exposure of the cells to pollutants (28), or induce expression of an analytically useful signal specifically and only in the presence of one or a limited set of structurally related compounds (1,14,20,26,32). For a number of reasons, it is useful to have microorganisms themselves monitor the immediate environment rather than using analytical chemistry. First, bacterial biosensors can be used for quick screening purposes, either for detecting the presence of toxic compounds in aqueous samples or for detecting a predefined set of target compounds for which the biosensors had been developed. Second, having microorganisms themselves "measure" the concentrations of pollutants is thought to be more representative for estimation of the bioavailable fraction and could solve some of the difficulties of extrapolating bioavailable concentrations from differential chemical extrac...

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mentioning

confidence: 99%

Measurement of Biologically Available Naphthalene in Gas and Aqueous Phases by Use of a Pseudomonas putida Biosensor

Werlen

Jaspers

Meer

2004

Appl Environ Microbiol

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“…We have found that the mutations fall into three categories: (1) those that are responsive to DNT but not TNT, (2) those that are responsive to TNT but not DNT, and (3) those that respond to both DNT and TNT. 4 Representative examples of each class are presently being analyzed for the gene sequence, and we anticipate that the results will demonstrate which area of the gene is vital for interaction with the explosive chemical.…”

Section: Methodsmentioning

confidence: 99%

“…We have published work on bacterial strains for detection of hydrocarbons (e.g., toluene, naphthalene) (1,4), mercury (both organic and inorganic) (6), and a variety of exotic bioreporters of environmental conditions (e.g. nutrient starvation, DNA damaging events).…”

Section: Introduction 2 In Traductionmentioning

confidence: 99%

Bioreporter bacteria for landmine detection

Burlage

Youngblood²,

Lamothe³

1998

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"The submitted manuscript has been authored by a contractor ofthe U.S. government under contract no. DE-AC05-960R22464. Accordingly, the U.S. government retains a nonexclusive, royalty-free license to publish or reproduce the published form of chis contribution, or allow others to do so, for U.S. Government purposes." 0 a 3 0 1 DISCLAIMERThis report was prepared as an account of work sponsored by an agency of the United States Government Neither the United States Government nor any agency thereof, nor any of their employees, makes any warranty, express or implied, or assumes any legal liability or responsibility for the accuracy, completeness, or usefulotss of any information, apparatus, product, or proass disclosed, or represents that its use would not infringe privately owned rights. Reference herein to any specific commercial product, process, or service by trade name, trademark, manufacturer, or otherwise docs not necessarily constitute or imply its endorsement, m mmendation, or favoring by the United States Government or any agency thereof. The views and opinions of authors expressed herein do not n-rily state or reflect those of the United States Government or.any agency thereof. AbstractLandmines (and other UXO) gradually leak explosive chemicals into the soil at significant concentrations. Bacteria, which have adapted to scavenge low concentrations of nutrients, can detect these explosive chemicals. Uptake of these chemicals results in the triggering of specific bacterial genes. We have created genetically recombinant bioreporter bacteria that detect small concentrations of energetic chemicals. These bacteria are genetically engineered to produce a bioluminescent signal when they contact specific explosives. A gene for a brightly fluorescent compound can be substituted for increased sensitivity. By finding the fluorescent bacteria, you find the landmine. Detection might be accomplished using stand-off illumination of the minefield and GPS technology, which would result in greatly reduced risk to the deminers. Bioreporter technology has been proven at the laboratory scale, and will be tested under field conditions in the near future.We have created a bacterial strain that detects sub-micromolar concentrations of oand p-nitrotoluene. Related bacterial strains were produced using standard laboratory protocols, and bioreporters of dinitrotoluene and trinitrotoluene were produced, screening for activity with the explosive compounds. Response time is dependent on the growth rate of the bacteria. Although full signal production may require several hours, the bacteria can be applied over vast areas and scanned quickly, producing an equivalent detection speed that is very fast. This technology may be applicable to other needs, such as locating buried explosives at military and ordnance/explosive manufacturing facilities.

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“…Unlike immuno-or nucleic acid-based assays, cell-based sensors provide general and physiologically relevant information about a wide variety of substances. Cells are optimal biological elements for generic detection as they contain a variety of enzymes, receptors, channels and pathways that enable them to respond dynamically to their environment and have been utilized for a variety of applications including routine toxicological analysis and assessment of environmental contamination (Applegate, 1998;Belkin 1998;Burlage 1994). Cell-based sensors fill an important threat assessment niche as they are designed to simultaneously detect large numbers of both known and unknown threats, characterize the functionality of those threats, and to some extent predict human performance upon exposure (Stenger et al, 2001, Pancrazio et al, 1999.…”

Section: Dna-based Microarrays By Dr Joanne Andreadis Us Nrlmentioning

confidence: 99%

Options for Development of Environmental Sentinel Biomonitor Systems for Real-Time Detection of Toxic Chemicals in Response to US Military Needs

Reuter¹,

Schalie²,

Shedd³

et al. 2002

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Public reporting burden for this collection of information is estimated to average 1 hour per response, including the time for reviewing instructions, searching existing data sources, gathering end maintaining the data needed, and completing and reviewing the collection of information. Send comments regarding this burden estimate or any other aspect of this collection of information, including suggestions for reducing this burden, to Washington Headquarters Services, Directorate for Information Operations and Reports, 1215 Jefferson Davis Highway, Suite 1204, Arlington, VA 22202-4302, and to the Office of Management and Budget,, Washington, DC 20503. AGENCY USE ONLY (Leave blank)2. REPORT DATE ABSTRACT (Maximum 200 words)This report summarizes the substance of a workshop on environmental sentinel biomonitor systems (ESB) held 26-28 Sept 2001 at the US Army Center for Environmental Health Research. The focus of the workshop was on development and field use of ESB systems as real-time early warning sensors for the presence of toxic industrial chemicals and toxic industrial materials (TICs and TIMs) which might present health hazards to exposed personnel. The scope of the workshop was limited to human health considerations and issues, and a number of military scenarios were discussed. SUBJECT TERMSEnvironmental sentinel biomonitoring, toxic industrial chemicals, toxic industrial materials, real-time detection, preventive medicine, force health protection, early warning sensor For this project, ESB systems are defined as:• Biologically-based systems (in vivo, in vitro, synthetic) and chemical sensors placed in the environment (in situ), either mobile or stationary, that can sense a biologically-significant event and provide relevant real-time data for use in risk assessment, mitigation, and/or management.• Real-time data are not limited to data available instantaneously to the user/decision maker. Data available within the time frame required by the military decision maker are considered real-time for this project. This is envisioned to typically be less than one hour.The overriding questions the project attempted to answer are: The focus was on the scientific, not the engineering, aspects of integrated ESB systems, achievable within five years (referred to as near-term) and those achievable within five to 10 years (referred to as far-term), with the potential to ultimately provide continuous, real-time monitoring of single known and unknown Toxic Industrial Chemicals/Toxic Industrial Materials (TICs/TIMs) and their complex mixtures under anticipated military field conditions and operational scenarios integrating both biological effects and chemical measurements and possibly triggering supplemental efforts (e.g. sample collection and/or additional analyses).The Workshop Agenda, Attendees' List and Workshop Charge are in Appendices A, B and C, respectively. The White Paper (Appendix D) was peer reviewed and copies distributed to all persons invited to the Workshop for comment. It has not been formally staffed. APP...

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Bioluminescent reporter bacteria detect contaminants in soil samples

Cited by 44 publications

References 6 publications

Measurement of Biologically Available Naphthalene in Gas and Aqueous Phases by Use of a Pseudomonas putida Biosensor

Measurement of Biologically Available Naphthalene in Gas and Aqueous Phases by Use of a Pseudomonas putida Biosensor

Bioreporter bacteria for landmine detection

Options for Development of Environmental Sentinel Biomonitor Systems for Real-Time Detection of Toxic Chemicals in Response to US Military Needs

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