Specification and Numerical Analysis of the Fluorescent Behavior of Luminescent Bacteria in Water Toxicity Testing
A technology for luminescent bacteria and toxicity detection, which is applied in fluorescence/phosphorescence, material excitation analysis, etc., can solve problems such as no literature records, achieve high reproducibility, and improve data reproducibility
Inactive Publication Date: 2016-09-14
刘星海
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[0012] There is currently no literature documenting how to normalize the fluorescence behavior of these luminescent bacteria to obtain a reliable toxicity assay data.
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[0023] Strains and supporting media and reagents
[0024] 1. Marine luminescent bacteria Vibrio fischeri (Aliivibrio fischeri ATCC700601)
[0025] Purchased from American Type Culture Collection
[0026] culture medium : Each 1 liter medium contains: 5 grams of yeast extract, 5 grams of tryptone, 0.5 grams of magnesium sulfate, 1 grams of calcium carbonate, 3 grams of potassium hydrogen phosphate, 30 grams of sodium chloride, if necessary, add 20 grams of agar medium agar. Store at room temperature after autoclaving at 121°C.
[0027] Cryoprotectant : Each 100 ml of frozen culture solution contains: 60 grams of glucose, 4 grams of sodium chloride, 2 grams of L-histidine, 0.5 grams of bovine serum albumin, fully dissolved and adjusted to a pH value of 7 for later use.
[0028] Lyoprotectant : Each 100ml lyophilized liquid contains: 8g skimmed milk powder, 8g glucose, 0.5g arginine and 3g sodium chloride, after fully dissolving, adjust the pH value to 7 for later use. ...
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Although toxicity can be well quantified by contacting luminous bacteria with toxic substances, the quantified toxicity value is difficult to use in the fields of scientific communication and actual detection and monitoring of water environment because of the biological diversity. The normalization and data analysis of fluorescence behavior of luminous bacteria in water toxicity detection greatly improve the reproducibility of toxicity detection utilizing luminous bacteria. If a same standard toxin and same luminous bacterium dosage form are used in detection, the error of detection result is less than 10%. For example, a freeze-dried aliivibrio fischeri strain is used to detect the toxicity of phenol (analytically pure), the toxicity detection result has a very high reproducibility, which is not affected by operators or detection locations, and the error of the detection result is less than 10%. So the normalization and data analysis can be used to evaluate water toxicity, and the results can be used for scientific communication. Through the long-term observation and repeated tests on luminous bacteria in lab, a series of methods that can greatly improve the reproducibility of toxicity detection utilizing luminous bacteria are developed, and the achievement mainly comprises the two inventions as follows: (1) a method for normalizing the fluorescence behavior of luminous bacteria; (2) a method for reducing the error through data analysis.
Description
[0001] Specification and Numerical Analysis of the Fluorescent Behavior of Luminescent Bacteria in Water Toxicity Testing 1. Technical field [0002] The present invention can quantify the ecotoxicity (ecotoxicity) of any unknown water-soluble toxin in a short time (one hour), these toxins include inorganic salts, organic compounds and colored water; can be done without any background in analytical chemistry Warning about water toxicity. To put it simply, I don't know, and I don't care what kind of water-soluble substance it is, but I want to know how toxic it is? What are the biological (ecological) consequences? [0003] The present invention is not suitable for the evaluation and judgment of the carcinogenicity of chemical substances or the danger of pathogens; it is also not suitable for the judgment of the toxic response of eukaryotic cell receptors, also known as pharmacology and toxicology. [0004] The present invention may be potentially misleading. For example, alc...
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IPC IPC(8): G01N21/64
Inventor 刘星海
Owner 刘星海