Photobacterium test

A bacterial assay, the Photobacterium test, based on the inhibition of light emission of a bioluminescent bacterium Vibrio fisheri (ISO, 1998), originally developed to test the toxicity of industrial effluents, gives an indication of the effects of the test agent on the oxidative metabolism of the cell. As a bacterial assay its advantages are rapidity and relatively low costs, but naturally the differences between bacterial and mammalian systems make the interpretation of the results even more difficult than with other short-term tests. 

So far there have been relatively few published reports on the application of in-vitro toxicological tests to extracts of paper and board. The published results relate to the use of photobacterium test, RNA-synthesis inhibition assay or to a battery of different test systems. The outcomes of these trials are summarised below. 

JOKINEN K, SAVOLAINEN M and soDERHJELM L (2001) Photobacterium test for evaluation of toxicity of fibrous products. Pap. Puu, 83, 332-335. 

The tests which supplied most of the data for Table V were those using 24-hour incubation at the temperature appropriate for each species of bacterium. The serial dilutions represent bacteriostatic concentrations and not necessarily bactericidal values With several of the antibiotics, activity decreases upon incubation beyond the 24 hours selected arbitrarily as the incubation time. The Photobacterium tests of Rake, Jones, and McKee (136) were read after 30 minutes, those of the author (101) after 24 hours. 

Kamlet, M. J., Doherty, R. M., Veith, G. D., Taft, R. W., Abraham, M. H. (1986) Solubility properties in polymers and biological media. 7. An analysis toxicant properties that influence inhibition of bioluminescence in Photobacterium phosphoreum (the Microtox test). Environ. Sci. Technol. 20, 690-695. 

The first steps in bypassing of the biological, technological, and financial burden of live stock culturing or maintenance were made more than 20 years ago through the development of a bacterial luminescence inhibition test [34,35] this bioassay is presently known and used worldwide as the Microtox test. The revolutionary principle of this test is that it uses a lyophihzed strain of a (marine) bacterium Photobacterium phosphoreum). This makes the bioassay apphcable anytime, anywhere, without the need for continuous culturing of the test species. 

Ribo, J.M. Kaiser, K.L.E. Photobacterium phosphoreum toxicity bioassay. I. Test procedures and application. Toxic. Assess. 1987, 2, 305-323. 

Moran, T. and Chiles, C. (1993) Multi-species toxicity assessment of sediments from the St-Clair River using Hyalella azteca, Daphnia magna and Microtox (Photobacterium phosphoreum) as test organisms, Canadian Technical Report of Fisheries and Aquatic Sciences 1942, 447-456. 

Environment Canada. (1992b) Biological test method toxicity test using luminescent bacteria (Photobacterium phosphoreum). Environmental Protection, Conservation and Protection, Environment Canada. Ottawa, Ontario, Reference Method EPS l/RM/24. 

Hermens J, Busser F, Leeuwangh P, Musch A. 1985a. Quantitative structure-activity relationships and mixture toxicity of organic chemicals in Photobacterium phosphoreum the Microtox test. Ecotoxicol Environ Safety 9 17-25. 

Some alternative ecotoxicological bioassays are available for environmental monitoring, and amongst these the tests based on invertebrates such us Daphnia magna (ISO, 1996), microalgae, such as Skeletonema costatum (ISO, 1995) and Selenastrum capricornotum (EPA, 1982), the marine bacteria Vibrio fischeri and Photobacterium phosphoreum (ISO, 1998) are well established. These tests use standardized organisms, and are available from a number of commercial companies. 

Luminescence test using Vibrio fischeri (formerly Photobacterium phosphor-eurri) (Environment Canada, 1992b). 

Luminous bacteria are bioluminescent microorganisms whose luciferase genes (lux), proteins and intact cells are widely used in applied research and commercial products. Acknowledging the commercial value of luminescent cells also in entertainment and education, we have conducted research on luminous bacteria from marine samples and have isolated Photobacterium phosphoreum (strain RL-1) from coastal marine sediment. In order to maximize the luminescence activity of RL-1, we examined a series of extracts prepared from dried marine foodstuff. Because chitinous compounds and some amino acids are known to be abundant in dried squid and shrimp, we also tested the effects of those compounds on the luminescence activity. Among the supplemental compounds tested, chitosan, cysteine, and aspartic acid were found to enhance the luminescence activity of RL-1. The present results indicate that some amino acids and chitinous compounds are effective supplements for further enhancing bacterial light production in an enriched medium (SWC ). 

We developed the first expert system that incorporates a working set of rules for a type of QSAR referred to as a linear solvation energy relationship or LSER (13-17) to predict LSER variable values from SMILES string formalism. The program also uses these LSER results and information about toxicity to predict acute toxicity to four representative organisms the fathead minnow (Pimephales promelas), the crustaceans Daphnia magna and Daphnia pulex. and Photobacterium phosphoreum. the luminescent agent in the Microtox test. 

A new in vitro genotoxicity assay has been developed by the Microbics Corp. This assay, known by the trade name of Mutatox assay, is a convenient method to detect DNA-damaging substances (genotoxins) by measuring light emissions from an isolated dark mutant strain of Photobacterium phosphoreum. The Mutatox assay compares favorably in sensitivity with the Ames test, and it is easier and more rapid to perform [15]. As a toxicity estimation method, one convenient assay method was developed and uses cultured fish cells this is introduced in section 7.4. 

Whale, G.F., Jarrill, F.M., and Bashford, V., Assessment of the Suitability of the Photobacterium phosphor eum (Microtox ) Test to Provide Marine Toxicity Data for Offshore Chemical Control Schemes, in Ecotoxicology Monitoring, Richardson, M.L. (Ed.), VCH Publishers, Weinheim, 1993, pp. 251-260. 

Bacterial bioassays have been also investigated to determine if they can provide simple routine methods for the analysis of cyanotoxins. The one that has received more attention is the Microtox bioluminescence assay, which indicates toxicity by a reduction of the light, emitted by the test bacterium (Photobacterium phosphoreum). Further analysis revealed that there is no correlation between response in the Microtox assay and cellular content of the known cyanotoxins [157]. 

The bioluminescence inhibition assay for acute toxicity based on the bacterium Vibrio Jischeri (formerly Photobacterium phosphoreum) is a standard eco-toxicological bioassay in Europe (DIN EN ISO 11348). Because this method is very rapid and cost-effective it has been widely used in environmental toxicity studies, e.g., routine screening of river and lake water or effluents, monitoring of bioremediated soils, or toxicity testing of chemicals [14]. 

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