A kind of denitrifying bacteria that tolerates high salt and efficiently degrades nitrate and its preparation and application
A technology of denitrifying bacteria and nitrate, 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, achieve efficient, stable and thorough treatment methods, and accelerate domestication and start-up efficiency , good economic benefits and environmental benefits
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Embodiment 1
[0055] Example 1: Isolation and identification of Pannonibacter phragmitetus
[0056] (1) enrichment, separation medium and expansion medium:
[0057] Enrichment medium: potassium nitrate 2g, magnesium sulfate heptahydrate 0.2g, dipotassium hydrogen phosphate 1.76g, potassium sodium tartrate 20g, ferrous sulfate 7hydrate 0.005g, calcium chloride 0.02g, ammonium chloride 0.63g, NaCl 5 %, distilled water 1000mL, pH value 7.5.
[0058] 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 Thymol blue solution.
[0059] Fermentation medium: peptone 8g, yeast extract 4g, sodium chloride 10g, distilled water 1000mL pH 7.5.
[0060] (2) Isolation and purification of Pannonibacter phragmitetus strain:
[0061] Weigh 5g of the acclimated denitrification sludge and put it in a 250mL anaerobic bottle, add ...
Embodiment 2
[0070] Example 2: Preparation of Pannonibacter phragmitetus inoculum
[0071] After the activation and cultivation of Pannonibacter phragmitetus, the inoculation amount of 5% (volume ratio) was inserted into the fermentation medium, and the fermentation culture conditions were controlled as follows: 35°C, dissolved oxygen 3 -10 4 The bacterial suspension is filled to form a liquid Pannonibacter phragmitetus bacterial agent product, which is used for the denitrification and denitrification treatment of high-salt and high-concentration nitrate waste liquid.
[0072] The composition of the fermentation medium is: peptone 10g / L, yeast extract 5g / L, sodium chloride 50g / L, the remainder is distilled water, the pH value is 7.5, sterilized at 121° C. for 20 minutes, and cooled to room temperature after sterilization.
Embodiment 3
[0073] Example 3: Preparation of inoculum of Pannonibacter phragmitetus
[0074] The Pannonibacter phragmitetus was inoculated into the seed medium, the culture temperature was controlled at 35° C., and the culture time was shaken at 40 rpm for 48 hours to obtain a seed fermentation broth.
[0075] The seed fermentation liquid is inserted into the fermentation medium according to the inoculation amount of 10% (volume ratio), and the fermentation culture conditions are controlled as follows: 35°C, dissolved oxygen <0.2mg / L, tank pressure: 0.03-0.10Mpa, 30-50rpm shaking culture Time 72h.
[0076] The preparation method of the bacterial agent is as follows: after the Pannonibacter phragmitetus is fermented, under the condition of 4° C., 7500 rpm, and centrifuged for 10 min, discard the supernatant to obtain the bacterial cell precipitate, and use the fermentation medium for the bacterial cell precipitate. Diluted to obtain bacterial viability of 10 3 The bacterial suspension of...
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