Preparation method and application of a kind of high-salt-tolerant denitrifying bacteria and its bacterial agent
A technology of nitrifying bacteria agent and denitrifying bacteria, which is applied in the field of environmental microorganisms, can solve the problems of incomplete treatment of high-concentration nitrate waste liquid, secondary pollution and salt tolerance, and complicated treatment process, and achieve efficient, stable and thorough treatment method, good economic benefit and environmental protection benefit, efficient and stable denitrification effect
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Embodiment 1
[0056] Embodiment 1: the isolation and identification of bacterial strain Pseudomonas stutzeri:
[0057] (1) Composition of enrichment, separation medium and expansion medium:
[0058] Enrichment medium: potassium nitrate 2g, magnesium sulfate heptahydrate 0.2g, dipotassium hydrogen phosphate 1.76g, potassium sodium tartrate 20g, ferrous sulfate heptahydrate 0.005g, calcium chloride 0.02g, ammonium chloride 0.63g, NaCl 5 %, distilled water 1000mL, pH value 7.5.
[0059] Separation medium: Potassium nitrate 2g, magnesium sulfate heptahydrate 0.2g, dipotassium hydrogen phosphate 0.5g, potassium sodium tartrate 20g, NaCl 5%, distilled water 1000mL, pH 7.0-7.2, solid medium plus agar 1.5% and 1mL bromine Musk thymol blue solution.
[0060] Fermentation medium: peptone 8g, yeast extract 4g, sodium chloride 10g, distilled water 1000mL pH value 7.5.
[0061] (2) Isolation and purification of Pseudomonas stutzeri:
[0062] Weigh 5g of the acclimatized denitrification sludge into a...
Embodiment 2
[0071] Embodiment 2: the preparation of Pseudomonas stutzeri bacterial agent
[0072] After Pseudomonas stutzeri is activated and cultivated, it is inserted into the fermentation medium according to the inoculation amount of 5%, and the fermentation temperature is controlled: 30°C, dissolved oxygen 3 -10 4 Cfu / L bacterial suspension, the bacterial suspension is filled to form a liquid Pseudomonas stutzeri agent product, which is used for the denitrification and denitrification treatment of high-salt and high-concentration nitrate waste liquid.
[0073] The composition of the fermentation medium is: peptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L, the balance is distilled water, pH 7.5, sterilized at 121°C for 20min, cooled to room temperature after sterilization.
Embodiment 3
[0074] Embodiment 3: the bacterial agent preparation of Pseudomonas stutzeri
[0075] The Pseudomonas stutzeri was inoculated into the seed culture medium, the culture temperature was controlled at 32° C., and the culture was shaken at 35 rpm for 52 hours to obtain a seed liquid.
[0076] Put the Pseudomonas stutzeri seed solution into the fermentation medium according to the inoculation amount of 10%. The fermentation culture conditions are: at 35°C, dissolved oxygen <0.2mg / L, tank pressure 0.03-0.10Mpa, and the culture time 72h;
[0077] Place the fermented liquid in a refrigerated centrifuge at 4°C, 7500rpm, centrifuge and concentrate for 10min, discard the supernatant, and dilute the precipitate with sterile saline with a sodium chloride concentration of 10g / L to obtain a bacterial activity of 10 3 Cfu / L bacterial suspension, the bacterial suspension is filled to form a liquid Pseudomonas stutzeri agent product, which is used for the denitrification and denitrification tr...
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