Indigenous high-efficiency denitrification strains of Pseudomonas fulva K3 and Pseudomonas mosselii K17 in soil of rare earth leaching site and application of strains

Abstract

The invention relates to indigenous high-efficiency denitrification strains of Pseudomonas fulva K3 and Pseudomonas mosselii K17 in soil of a rare earth leaching site and application of the strains. The strains are both enriched and screened from the soil of rare earth leaching site, and belong to Pseudomonas fulva and Pseudomonas mosselii respectively, both of the strains are transferred to ChinaCenter for Type Culture Collection for preservation on Decomber 16, 2019, and the corresponding deposit numbers are respectively CCTCC NO: M 20191055 and CCTCC NO: M 20191056. The two strains are both heterotrophic nitrifying-aerobic denitrifying bacteria, can quickly remove ammonia nitrogen in a leachate (initial ammonia nitrogen concentration <= 300 mg / L)from the rare earth leaching site underaerobic conditions, the denitrification rate is more than 90%, the accumulation of nitrite nitrogen and nitrate nitrogen is less than 6 mg / L, and the ammonia nitrogen concentration in the leachate after treatment reaches the national emission standard. The discovery of the strains provides a high-efficiency and environment-friendly microbial denitrification treatment method for the lechate from the rare earth leaching site, and the strains have broad application prospects.

Description

technical field

[0001] The invention relates to the technical field of microbes and mine sewage treatment, in particular to the native high-efficiency denitrification bacterial strains Pseudomonas chrysogenum K3 and Pseudomonas morgii K17 in the soil of a rare earth leaching site and applications thereof. Background technique

[0002] The weathering crust elution-type rare earth ore is an important strategic resource in my country. The content of medium and heavy rare earth in this kind of rare earth ore is relatively high, and it has a great competitive advantage in the international market. Weathering crust leaching-type rare earth ores have been discovered in seven provinces in southern my country and have been mined and utilized. The mining process has also developed from pool leaching to heap leaching and then to the current in-situ leaching. Ammonium sulfate is the most widely used leaching agent in the in-situ leaching process of weathering crust leaching type rare ea...

Examples

Embodiment 1

[0041] Inoculate the native high-efficiency denitrification bacterial strain K3 in the soil of the rare earth leaching site stored on the slant of the test tube at 4°C to a plate containing a solid medium for denitrification (see 3-(2) for the preparation method of the plate), and cultivate at 28°C 12h, to achieve strain activation. Under sterile conditions, use an inoculation loop to inoculate the activated strain into a 250mL Erlenmeyer flask filled with 100mL of denitrification liquid medium, place the Erlenmeyer flask in a shaker and culture it for 12 hours at 28°C and 165r / min. , to obtain the seed solution. Based on the denitrification liquid medium, the initial concentration of ammonia nitrogen was adjusted to 100 mg / L, and the C / N ratio was 12 to obtain a simulated leachate from a rare earth leaching site. The seed solution was inoculated into 100mL of the sterilized simulated leachate from the rare earth leaching site with 2% inoculum amount, and the shaking culture ...

Embodiment 2

[0044] Seed liquid was prepared with reference to the method of Example 1. Based on the denitrification liquid medium, the initial concentration of ammonia nitrogen was adjusted to 200 mg / L, and the C / N ratio was 12 to obtain a simulated leachate from a rare earth leaching site. The seed solution was inoculated into 100mL of the sterilized simulated leachate from the rare earth leaching site with 2% inoculum amount, and the shaking culture was carried out under the conditions of 28°C and 165r / min, and samples were taken regularly to measure the ammonia nitrogen content, OD600, Nitrite nitrogen content, nitrate nitrogen content, drawn to get Figure 4 .

[0045] Depend on Figure 4 It can be seen that the concentration of ammonia nitrogen in the simulated leachate of the rare earth leaching site dropped from the initial 200mg / L to 1.4mg / L after 24 hours of inoculation seed solution treatment, and the calculation shows that the removal rate of ammonia nitrogen is as high as 99...

Embodiment 3

[0047] Seed liquid was prepared with reference to the method of Example 1. Based on the denitrification liquid medium, the initial concentration of ammonia nitrogen was adjusted to 300 mg / L, and the C / N ratio was 12 to obtain a simulated leachate from a rare earth leaching site. The seed solution was inoculated into 100mL of the sterilized simulated leachate from the rare earth leaching site with 2% inoculum amount, and the shaking culture was carried out under the conditions of 28°C and 165r / min, and samples were taken regularly to measure the ammonia nitrogen content, OD600, Nitrite nitrogen content, nitrate nitrogen content, drawn to get Figure 5 .

[0048] Depend on Figure 5 It can be seen that the ammonia nitrogen concentration in the simulated leachate of the rare earth leaching site dropped from the initial 300 mg / L to 12.37 mg / L after 48 hours of inoculation seed solution treatment, and the calculation shows that the ammonia nitrogen removal rate is as high as 95.8...

technical field

[0001] The invention relates to the technical field of microbes and mine sewage treatment, in particular to the native high-efficiency denitrification bacterial strains Pseudomonas chrysogenum K3 and Pseudomonas morgii K17 in the soil of a rare earth leaching site and applications thereof. Background technique

[0002] The weathering crust elution-type rare earth ore is an important strategic resource in my country. The content of medium and heavy rare earth in this kind of rare earth ore is relatively high, and it has a great competitive advantage in the international market. Weathering crust leaching-type rare earth ores have been discovered in seven provinces in southern my country and have been mined and utilized. The mining process has also developed from pool leaching to heap leaching and then to the current in-situ leaching. Ammonium sulfate is the most widely used leaching agent in the in-situ leaching process of weathering crust leaching type rare ea...

Examples

Embodiment 1

[0041] Inoculate the native high-efficiency denitrification bacterial strain K3 in the soil of the rare earth leaching site stored on the slant of the test tube at 4°C to a plate containing a solid medium for denitrification (see 3-(2) for the preparation method of the plate), and cultivate at 28°C 12h, to achieve strain activation. Under sterile conditions, use an inoculation loop to inoculate the activated strain into a 250mL Erlenmeyer flask filled with 100mL of denitrification liquid medium, place the Erlenmeyer flask in a shaker and culture it for 12 hours at 28°C and 165r / min. , to obtain the seed solution. Based on the denitrification liquid medium, the initial concentration of ammonia nitrogen was adjusted to 100 mg / L, and the C / N ratio was 12 to obtain a simulated leachate from a rare earth leaching site. The seed solution was inoculated into 100mL of the sterilized simulated leachate from the rare earth leaching site with 2% inoculum amount, and the shaking culture ...

Embodiment 2

[0044] Seed liquid was prepared with reference to the method of Example 1. Based on the denitrification liquid medium, the initial concentration of ammonia nitrogen was adjusted to 200 mg / L, and the C / N ratio was 12 to obtain a simulated leachate from a rare earth leaching site. The seed solution was inoculated into 100mL of the sterilized simulated leachate from the rare earth leaching site with 2% inoculum amount, and the shaking culture was carried out under the conditions of 28°C and 165r / min, and samples were taken regularly to measure the ammonia nitrogen content, OD600, Nitrite nitrogen content, nitrate nitrogen content, drawn to get Figure 4 .

[0045] Depend on Figure 4 It can be seen that the concentration of ammonia nitrogen in the simulated leachate of the rare earth leaching site dropped from the initial 200mg / L to 1.4mg / L after 24 hours of inoculation seed solution treatment, and the calculation shows that the removal rate of ammonia nitrogen is as high as 99...

Embodiment 3

[0047] Seed liquid was prepared with reference to the method of Example 1. Based on the denitrification liquid medium, the initial concentration of ammonia nitrogen was adjusted to 300 mg / L, and the C / N ratio was 12 to obtain a simulated leachate from a rare earth leaching site. The seed solution was inoculated into 100mL of the sterilized simulated leachate from the rare earth leaching site with 2% inoculum amount, and the shaking culture was carried out under the conditions of 28°C and 165r / min, and samples were taken regularly to measure the ammonia nitrogen content, OD600, Nitrite nitrogen content, nitrate nitrogen content, drawn to get Figure 5 .

[0048] Depend on Figure 5 It can be seen that the ammonia nitrogen concentration in the simulated leachate of the rare earth leaching site dropped from the initial 300 mg / L to 12.37 mg / L after 48 hours of inoculation seed solution treatment, and the calculation shows that the ammonia nitrogen removal rate is as high as 95.8...

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