NL2032165B1

NL2032165B1 - A preparation method of compound microbial agent

Info

Publication number: NL2032165B1
Application number: NL2032165A
Authority: NL
Inventors: He Da; Ke Leqin; Weng Jiaxue; Wu Min; Yan Zaixian; Zhu Yushuang; Zhu Zhenya; Liu Ying
Original assignee: Hangzhou Vocational & Technical College
Priority date: 2022-06-14
Filing date: 2022-06-14
Publication date: 2023-02-24

Abstract

The present invention discloses a compound microbial agent, which is composed of Bacillus pumilus, Bacillus licheniformis, Aspergillus oryzae and Bacillus amyloliquefaciens. According to the weight portions, include 40-50 portions of Bacillus pumilus, 40-50 portions of Bacillus licheniformis, 1-10 portions of Aspergillus oryzae and 1-10 portions of Bacillus amyloliquefaciens.

Description

A PREPARATION METHOD OF COMPOUND MICROBIAL AGENT

Technical Field

The present invention relates to a compound microbial agent, a preparation method and its application in sewage treatment,

Background Art

At present, the treatment method of domestic sewage is basically discharged to the sewage treatment plant through underground pipelines for treatment. However, the construction and operation of the sewage treatment plant not only costs a lot of money, but also the activated sludge method is not efficient in treating domestic sewage.

In order to achieve the purpose of degradation of pollutants in sewage, microbial agents are selected, cultivated and combined to form microbial communities with special degradation ability for sewage, and become special sewage treatment bacteria. This is one of the most advanced methods in sewage treatment technology at present. Microbial treatment technology is unique in sewage treatment, with low cost, complete purification, environmental protection, no pollution and simple operation. Microbial agents use the metabolism of microorganisms to purify sewage. A large amount of organic matter in sewage can be used for the growth and reproduction of microorganisms.

Generally, microbial agents do not form a good growth system in time after being put into use. Therefore, after sewage treatment for a period of time, the treatment effect will be reduced, and new microbial agents need to be added.

Detailed Description of the Invention

In order to overcome the shortcomings of the prior art, the present invention provides a compound microbial agent, a preparation method and its application in sewage treatment. The compound microbial agent can significantly reduce the concentration of COD in sewage, and can reduce the concentration of total nitrogen and total phosphorus in sewage. It can be used continuously for many times and reduce the cost of sewage treatment.

In order to achieve the above purpose, the present invention adopts the following technical scheme:

The present invention also discloses a preparation method of the compound microbial agent, which comprises the following steps:

(1) Respectively scraped one loop of lawn from the Bacillus pumilus, Bacillus licheniformis and Bacillus amylolyticus preserved on the slope with inoculation loop, inoculated them respectively into first liquid culture medium for culture at 37 °C for 1-2 days; (2) Scraped one loop of lawn from the Aspergillus oryzae preserved on the slope with inoculation loop, inoculated them into second liquid culture medium for culture at 28 °C for 1-2 days; (3) Centrifuge the culture solution of each bacterial strain obtained in steps (1) and (2) at 6000-8000r / min for 10-20min respectively, then wash with sterile water, filter and collect them respectively; {4) Freeze the bacteria collected in step (3) at - 20 °C for 10-12h, and then freeze-dry at - 50-- 60 °C for 12-30h; (5) Weigh and take 40-50 portions of Bacillus pumilus, 40-50 portions of Bacillus licheniformis, 1-10 portions of Aspergillus oryzae and 1-10 portions of Bacillus amyloliquefaciens according to the weight portions of the bacteria dried in step (4), and mix them to prepare a compound microbial agent;

The first liquid culture medium described in step {1} is composed of the following components: 3 portions of bovine serum, 10 portions of peptone, 5 portions of sodium chloride and 1000 portions of water, and the above are the weight portions; Adjust the pH to 7.4-7.6.

The second liquid culture medium described in step (2) is composed of the following components: 1 portion of potassium dihydrogen phosphate, 0.5 portion of magnesium sulfate, 5 portions of peptone, 10 portions of glucose, 1000 portions of water and 3.5-4 portions of 1%

Bengal red aqueous solution, the above are the weight portions; Adjust the pH to 7.4-7.6.

Detailed Description of Embodiments

In order to enable those in the technical field to better understand the scheme of the present invention, the technical scheme of the present invention will be clearly and completely described below in combination with specific embodiments. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them.

Embodiment 1, wherein the said Bacillus thuloopiensis is selected from the strain with the preservation number of CGMCC No.16881. The strain is named Bacillus thuloopiensis, and the preservation date is June 10, 2007. The preservation unit is Institute of Microbiology, Chinese

Academy of Sciences, and the preservation address is China Beijing NO.1 Beichen West Road,

Chaoyang District, Beijing 100101;

The said Bacillus licheniformis is selected from the strain with the preservation number of

CGMCC No.1430. The strain is named Bacillus licheniformis. The preservation date is december 7, 1970. The preservation unit is Institute of Microbiology, Chinese Academy of Sciences, and the preservation address is China. Beijing NO.1 Beichen West Road, Chaoyang District, Beijing 100101;

The said Rhodotorula oryzae yeast is selected from the strain with the preservation number of CGMCC No.22363. The strain is named Rhodotorula oryzae. The preservation date is july 1, 2002. The preservation unit is Institute of Microbiology, Chinese Academy of Sciences, and the preservation address is China. Beijing NO.1 Beichen West Road, Chaoyang District,

Beijing 100101;

The said Bacillus amyloliquefaciens is selected from the strain with the preservation number of CGMCC No.11099. The strain is named Bacillus amyloliquefaciens. The preservation date is October 1, 1979. The preservation unit is the Institute of Microbiology, Chinese Academy of Sciences, and the preservation address is China Beijing NO.1 Beichen West Road, Chaoyang

District, Beijing 100101.

Embodiment 1

The preparation of bacteria includes the following steps: (1) Respectively scraped one loop of lawn from the Bacillus thuloopiensis, Bacillus licheniformis and Bacillus amylolyticus preserved on the slope with inoculation loop, inoculated them respectively into first liquid culture medium for culture at 37 °C for 1-2 days; (2) Scraped one loop of lawn from the Rhodotorula oryzae preserved on the slope with inoculation loop, inoculated them into second liquid culture medium for culture at 28 °C for 1-2 days; 23 (3) Centrifuge the culture solution of each bacterial strain obtained in steps (1) and (2) at 8000r / min for 15 min respectively, then wash with sterile water, filter and collect them respectively; (4) Freeze the bacteria collected in step (3) at - 20 °C for 10h, and then freeze-dry at - 55 °C for 24h, and collect the dried bacteria respectively.

The first liquid culture medium described in step {1} is composed of the following components: 3g bovine serum, 10.0g peptone, 5.0g sodium chloride and 1000 mL water, and the above are the weight portions; Adjust the pH to 7.4-7.6 with 1mol / L NaOH or 1mol / L HCI.

The second liquid culture medium described in step (2) is composed of the following components: 1.0g potassium dihydrogen phosphate, 0.5g magnesium sulfate, 5.0g peptone, 10.0g glucose, 1000mL water and 3.5mL 1% Bengal red aqueous solution; Adjust the pH to 7.4-7.6 with 1mol/ L NaOH or 1mol / L HCI.

Embodiments 2-4 and comparative examples 1-4

According to the weight portions of each bacteria shown in Table 1, weigh the bacteria prepared in embodiment 1 respectively, and then mix to prepare a compound microbial agent.

Table 1 Weight portions of each bacteria in embodiments 2-4 and comparative examples 1-4

Bacillus Bacillus Bacillus

Rhodotorula thuloopiensis licheniformis / amylofiquefaciens / oryzae / pcs) / pels) pc(s) pels)

Comparative 40 40 example 1

Comparative 40 40 1 example 2

Comparative 40 1 1 example 3

Comparative 40 1 1 example 4

Result:

Table 1-2: Treat domestic sewage respectively with the compound microbial agent prepared in embodiments 2-4 and comparative examples 1-4, in which the concentration change of total phosphorus (mg / L}

Comp Compar Compar

Comparati arative ative ative Embodi Embodi Embodi

Days ve examp example example ment 4 ment 3 ment 2 example 4 lel 2 3 [ew [ee [ew

Ew |e fe

It can be seen from Table 1-2 that this compound microbial agent can remove the total phosphorus in domestic sewage, with a removal rate of more 94%, and the treatment efficiency of total phosphorus is significantly higher than that of total nitrogen. The four kinds of bacteria in the compound microbial agent of the present invention work together and have a 5 high treatment capacity for total phosphorus in sewage.

Treat the sewage with the compound microbial agents prepared in comparative example 3 and comparative example 4, and after 5 days, the total phosphorus concentration also continued to be decreased, further indicating that Bacillus amylosus can promote the growth of other strains.

To sum up, Bacillus thuloopiensis, Bacillus licheniformis, Rhodotorula oryzae and Bacillus amylolyticus in the compound microbial agent have a strong ability to treat total phosphorus in sewage.

Claims

CONCLUSION

1. Method for making a composite microbial preparation, characterized in that it comprises the following steps: {1} Sheep 1 ring moss with an inoculation ring from Bacillus subtilis, Bacillus licheniformis and Bacillus amyloliguefaciens preserved on slant in a liquid medium a to culture them for 1-2 days in an environment with a temperature of 37°C; (2) put sheep 1 ring moss with an inoculation ring from Aspergillus oryzae preserved on slant in a liquid medium b to culture them in an environment with a temperature of 28°C for 1-2 days; {3} Centrifuge all stock cultures obtained in steps (1) and {2} at 6000-8000r/min for 10-20min, then wash with sterile water and filter and collect them separately; (4) freezing the organisms collected in step (3) at -20°C for 10-12 hours, and then freeze-drying them at -50 to -60°C for 12-30 hours; (5) Weigh the dried organisms in step (4) and count them according to the weights: 40-50 parts Bacillus subtilis, 40-50 parts Bacillus licheniformis, 1-10 parts Aspergillus oryzae and 1-10 parts Bacillus amyloliquefaciens, and mixing to obtain a composite microbial preparation; wherein the liquid medium a of step (1) consists of the following parts: 3 parts of bovine serum, 10 parts of peptone, 5 parts of sodium chloride and 1000 parts of water, the above parts being counted by weight; the pH is adjusted to 7.4-7.6; the liquid medium b from step (2) consists of the following parts: 1 part potassium dihydrogen phosphate, 0.5 part magnesium sulfate, 5 parts peptone, 10 parts glucose, 1000 parts water, 3.5-4 parts of 1% Bengal red water solution ; the above parts are counted by weight; the pH is adjusted to 7.4-7.6.