Bioluminescent bioassays

Curtis, C., Lima, A., Lozano, S.J. and Veith, G.D. (1982) Evaluation of a bacterial bioluminescence bioassay as a method for predicting acute toxicity of organic chemicals to fish. In Aquatic Toxicology and Hazard Assessment Fifth Conference, ASTM STP 766, Pearson, J.G., Foster, R.B. and Bishop, W.E. (eds), pp. 170-178. American Society for Testing and Materials, Philadelphia, PA. 

Kuznetsov A, Tyulkova N, Kratasyuk V, Abakumova V, Rodicheva E. The investigation of properties of chemicals for bioluminescent bioassays. Siberian ecological journal (Russian) 1997 5 459-65. 

Bioluminescence bioassays based on luminous bacteria and coupled enzyme system NADH-FMN-oxidoreductase-luciferase were adapted for monitoring the saline-water conditions of Lake Shira (Khakasia, Siberia). The differences in bioluminescence responses have been found to be related to the salt composition and the oxidation-reduction properties of water. Bioluminescent kinetics parameters, which are mostly sensitive to pollution under conditions of saline water, have been observed. Figure 1 shows the typical bioluminescence kinetics of the samples of water due to anthropogenic influence (beach) and control clear water (non-recreational area). 

Ecotoxicity assessment of water samples was carried out, in parallel to chemical analysis, using three standardized bioassays based on the micro-crustacean Daphnia magna, the algae Pseudokirchneriella subcapitata, and the bioluminescent bacteria Vibrio fischeri. 

Gustavson, K.E., Svenson, A., and Harkin, J.M. Comparison of toxicities and mechanism of action of / -alkanols in the submitochondrial particle and the Vibrio fischeri bioluminescence (Microtox ) bioassay. Environ. Toxicol. Chem., 17(10) 1917-1921, 1998. 

A. Firefly Bioluminescence as a Prototype for a Dioxetane-based Triggered Chemiluminescence (CIEEL) in Bioassays... 

At the time of its conception, the PEEP index integrated the results of a selection of practical small-scale screening bioassays which included the Vibrio fischeri bioluminescence inhibition test, the Selenastrum capricornutum growth inhibition... 

Nowadays, Microtox is the most popular bioassay available that uses bioluminescent bacteria as its active element. Analysts find it a useful tool for assessing pollution in different compartments of the environment, possessing as it does both the advantages of bioindicator techniques and the precision of classic instrumental analysis. Here are some examples of its application ... 

Industrial effluents Organic compounds chloro- and nitrophenols, nonionic surfactants, linear alkylated benzene sulfonates, benzene and naphthalene sulfonates, estradiol, ethinyl estradiol V. fischeri (ToxAlert 10, ToxAlert 100) There were certain correlations between the results of the chemical analyses and bioassays to a large extent, however, the composition of the samples remained unknown Distinct correlations were found between the inhibition of bioluminescence and compound content, but only in the case of nonylphenol, nonylphenol carboxylate, nonylphenol ethoxylate, and chlorophenols 84... 

CIEEL is of particular interest for the development of modern chemiluminescent bioassays. The most popular clinical bioassays utilize thermally persistent spiro-adamantyl-substituted dioxetanes with a protected phenolate moiety. These designed 1,2-dioxetanes include an energy source, a fluorophore, and a trigger grouping, and are therefore structurally similar to bioluminescent substrates such as firefly luciferin. Three main commercial dioxetanes 75 are available as one-reagent assays for alkaline phosphatase and are sold under the name of AMPPD (R1 = R2 = H), CSPD (R1 = Cl, R2 = H), and CDP-Star (R1 = R2 = Cl) <2006S1781, 2003ANA279>. These substrates are sensitive to 10 21 mol of alkaline phosphatase in solution. 

One of the most important quality characteristics of advanced oxidation processes is their ability to reduce the toxicity of an industrial wastewater and to enhance its biodegradability (see Chapter 7.1.5). This criterion can be established by several standardized procedures using different test organisms ranging from microbes to intact animals (cf Tab. 5-1). An easy to perform variant is the bioluminescence assay that uses the inhibition of the bioluminescence intensity of the test organism Vibrio fischeri in the presence of toxic substrates. This is a bioassay used worldwide for the evaluation of toxicity data of individual chemicals or of industrial wastewaters (Froehner et al, 2000, DIN, 1991). Commercial systems are... 

The inhibition of light emitted by the bioluminescent bacterium is the basis for different toxicity bioassays. High sensitivity of the luminescent system even to micro-quantities of toxicants, rapid attainment of results, and exact quantification of... 

Bacterial luciferase coimmobilized with NAD(P)H FMN oxidoreductase on starch gel has been used for bioluminescent assay of aldehydesCo-immobilization of bacterial luciferase, NAD(P)H FMN oxidoreductase and their substrates is referred to as multifunctional immobilized biosensor and is a new trend for use of bioluminescent analysis, e.g. toxicity biotest and bioassay. The main principle of this luciferase biotest is the correlation between toxicity of the sample being studied and changes in bioluminescence parameters in vitro. Toxicity of the sample is measured by the changes in bioliuninescence intensity compared with that of a control. Multifunctional immobilized biosensors based on luciferase have been used for the following bioassays. 

The effect of blood serum, lymph and other biological liquids on bioluminescent reactions has been studied. The immobilized luciferase shows lower sensitivity than the soluble enzyme in human clinical tests, and in analysis of com and bread infection by fungi. But this problem is overcome by using larger samples, and hence this new biosensor can be successfully used for toxicity bioassays. 

To estimate water quality, bioluminescent biosensors have been devised and successfully used. They are characterized by rapidity and simplicity of use, high sensitivity, and accuracy. The Collection of Luminous Bacteria IBSO (http //www.bdt.org.brA3dt/msdn/ibso) is being used to develop bioassays for monitoring the environment, using lyophilized luminous bacteria and the luminescent system isolated from them. Bioluminescent assays have an advantage over other biological assays luminescence is easy to measure, the method is rapid, and the measurements can be automated. 

Weitz H J, Campbell C D, Killham K. Development of a novel, bioluminescence-based fungal bioassay for toxicity testing. Environ Microbiol 2002 4 422-9. 

Coelenterazine, a bioluminescent compound from a jellyfish, with potential for use in bioassays, has been synthesised in an overall 25% yield from chloropyrazine. ... 

Bioassays have been widely used for the evaluation of potentially contaminated sites - with varying degrees of success. For test applications, it is crucial to determine in advance what information is expected from the test and how the test data have to be processed in order to obtain this information. The percentage of plant growth reduction and bioluminescence inhibition of two bioassays, for example, will not be conclusive for remediation needs but may indicate the presence of pollution and the necessity for further investigation. 

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