Researches on Improving Sensitivity of Luminous Bacteria Toxicity Test Using Chelating Agent

Abstract:

The biological toxicity test of luminous bacteria has been widely applied. However, sensitivity of this test needs to be further improved. Cell membranes are bacteria’s first barrier against outside invasions. In this research, permeability of cell membranes will be changed so as to improve toxicity detection sensitivity of vibrio qinghaiensis (Q67). During the cultivation process of Q67, the Ethylene Diamine Tetraacetic Acid (EDTA-Na₂) will be added and the hydrophobic fluorescent probe N-Phenyl-1-Naphthylamine (NPN) will be used to test changes of Q67 cell permeability. In addition, the low temperature centrifuge method will be used to eliminate interferences of residual EDTA of the nutrient medium to the acute toxicity test. The microporous plate method of biological toxicity test will be used to test the acute toxicity of K₂Cr₂Q₇, the phenol and the desmetryn on Q67 before and after the permeability is modified. The results show that: during the cultivation process of Q67, if 2.0 mmol/L EDTA-Na₂ is added, 40.2% of its absorbability to NPN can be improved. What is more, normal growth and bioluminescent property of Q67 will not be affected. EC₅₀ of the K₂Cr₂Q₇, the phenol and the desmetryn to the modified Q67 is 4.39 mg/L, 128.75 mg/L and 145.89 mg/L respectively. Compared with those before the modification, their EC₅₀ reduces 45.70%, 39.77% and 38.61% respectively. Thus, during the cultivation process of Q67, the addition of EDTA-Na₂ can improve its permeability to poisonous substances and enhance the sensitivity of Q67 biological toxicity test.

• Periodical: Advanced Materials Research (Volumes 1030-1032)
• Pages: 2390-2396
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.1030-1032.2390
• Online since: September 2014
• Authors: Na Wang*, Xiao Chang Wang, Xiao Yan Ma
• Keywords: Acute Toxicity Test, EDTA-Na₂, Permeability, Sensitivity, Vibrio qinghaiensis sp. Q67

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