Improved nitrobacteria Diphenylamine-Griess detection method
A technology of nitrifying bacteria and detection methods, applied in the field of environmental chemistry and environmental microorganisms, can solve problems such as residues, and achieve the effects of improving reaction rate and conversion rate, and improving accuracy and timeliness.
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[0017] 1. Preparation of culture medium
[0018] (1) Liquid culture medium for nitrifying bacteria: 1.0g NaNO 2 , 1.0g Na 2 CO 3 , 0.25g NaH 2 PO 4 , 1.0g CaCO 3 , 0.75g K 2 HPO 4 , 0.01g MnSO 4 , 0.03g MgSO 4 4H 2 O, 1000mL distilled water, pH7.2; sterilize at 121°C for 30min.
[0019] (2) Enrichment culture: According to the formula of the culture medium, weigh the drug, prepare 1000ml culture medium in the Erlenmeyer flask, adjust the pH to 7.2, and then divide it into test tubes, 90ml, 121°C, autoclave sterilization 30min. Put 10 mL of the collected mud sample into a previously sterilized Erlenmeyer flask containing glass beads and 90 mL of culture medium, and culture it on a shaker at 15°C at 160 r / min for 10 days. The purpose is to break up the bacteria micelles, make the bacteria dispersed in the water in a single-cell state, and at the same time increase the number of nitrifying bacteria in the mud sample.
[0020] (3) Preparation of nitrifying bacteria cu...
Embodiment 2
[0024] The difference from Example 1 is:
[0025] First add 5-8 drops of hydrochloric acid to acidify the culture solution, then add 25-27 ammonium sulfamate, and react at 55-60°C until the color of the medium is colorless and there is gas at the bottom of the culture solution escape, and when the degassing stops, add a grain of ammonium sulfamate to make NO 2 - fully converted to N 2 And escape, take the culture solution on the white porcelain color comparison plate, add 3-5 drops of Griess reagent, the culture solution is colorless, finally take the culture solution on the color comparison plate, add 3-5 drops of diphenylamine, the culture solution Appears blue.
[0026] The cultivation of the nitrifying bacteria is to cultivate the nitrifying bacteria in the nitrifying bacteria culture solution for 15 days at 20° C. and 150 r / min, break up the bacteria micelles, and disperse the bacteria into a single-cell state in water, while increasing the number of nitrifying bacteri...
Embodiment 3
[0028] The difference from Example 1 is:
[0029] First add 5-8 drops of hydrochloric acid to acidify the culture solution, then add 28-30 ammonium sulfamate, and react at 60°C until the color of the medium is colorless, and gas escapes from the bottom of the culture solution , when the degassing stops, add a grain of ammonium sulfamate to make NO 2 - fully converted to N 2And escape, take the culture solution on the white porcelain color comparison plate, add 3-5 drops of Griess reagent, the culture solution is colorless, finally take the culture solution on the color comparison plate, add 3-5 drops of diphenylamine, the culture solution Appears blue.
[0030] The cultivation of the nitrifying bacteria is to cultivate the nitrifying bacteria in the nitrifying bacteria culture solution for 13 days at 30° C. and 155 r / min, break up the bacteria micelles, disperse the bacteria in the water in a single cell state, and simultaneously increase the number of nitrifying bacteria ...
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