Preparation method and application of a kind of salt-tolerant denitrifying bacteria and bacterial agent
A denitrifying bacteria and denitrification technology, 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 effect, etc., achieve high-efficiency, stable and thorough treatment methods, and speed up domestication and start-up efficiency. , the effect of strong tolerance
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Embodiment 1
[0058] Embodiment 1: the isolation identification of bacterial strain of the present invention:
[0059] (1) Enrichment, separation medium and expansion medium:
[0060] 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.
[0061] 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.
[0062] Fermentation medium: peptone 8g, yeast extract 4g, sodium chloride 10g, distilled water 1000mL pH value 7.5.
[0063] (2) Isolation and purification of Halomonas bacterial strain:
[0064] Weigh 5g of the acclimatized denitrification sludge into a 250mL an...
Embodiment 2
[0073] Embodiment 2: the preparation of halomonas bacterial agent
[0074] After the Halomonas was activated and cultivated, the Halomonas seed liquid was inserted into the fermentation medium according to the inoculation amount of 5%, and the fermentation temperature was controlled at 35°C, dissolved oxygen3 -10 4 cfu / L of the bacterial suspension, and fill it to finally obtain the halomonas microbial agent.
[0075] The composition of the fermentation medium is: peptone 10g / L, yeast extract 4g / L, sodium chloride 35g / L, the balance is distilled water, pH value 7.5, sterilized at 121°C for 20min, cooled to room temperature after sterilization.
Embodiment 3
[0076] Embodiment 3: the preparation of halomonas bacterial agent
[0077] The salt-tolerant denitrifying bacteria were inoculated into the seed culture medium, the culture temperature was controlled at 35° C., and the culture was shaken at 35 rpm for 48 hours to obtain a seed fermentation liquid.
[0078] Put the halomonas seed fermentation liquid into the fermentation medium according to the inoculum amount of 10%, control the fermentation temperature at 35°C, dissolve oxygen <0.2mg / L, tank pressure 0.03-0.10Mpa, and shake at 40rpm for 72h.
[0079] After the Halomonas was cultured by fermentation, centrifuge at 7500rpm for 10min at 4°C, discard the supernatant to obtain a bacterial cell precipitate, dilute the bacterial cell precipitate with a fermentation medium, and obtain a bacterial activity of 10 3 cfu / L of the bacterial suspension, and fill it to finally obtain the halomonas microbial agent.
[0080] The seed medium consists of: KNO 3 2g / L, MgSO 4 ·7H 2 O 0.2g / L,...
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