Mixed microbial preparation and manufacture method thereof
A technology of mixed microorganisms and manufacturing methods, which is applied in the field of mixed microbial preparations and its manufacturing, and can solve problems such as difficult effects, difficult recovery of oil-contaminated soil, and difficult effective treatment of a wide range of contaminated areas.
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no. 1 example
[0041] Experimental Example 1: Manufacture of a mixed microbial preparation with an oil-decomposing ability to achieve an optimal microbial composition ratio
[0042] First, the measurement of the decomposition rate using a biochemical oxygen demand (BOD, biochemical oxygen demand) intelligent coagulation test mixer (jar tester) will be described below.
[0043] figure 1 It is a figure explaining the manufacturing method of the mixed microorganism preparation of the Example of this invention.
[0044] refer to figure 1 , Accurately measured 160 soils where the dry soil was polluted by diesel oil (Diesel) at 10000ppm (mg / kg). After diluting the above-mentioned microorganisms and adding the moisture to 20% (step S101), use tweezers to put NaOH (Sodium hydroxide) in the rubber bag of the tablets to make the biochemical oxygen demand intelligent coagulation test mixer (OxiTOP) was completely sealed, and placed in a culture medium at 20° C. for zero calibration, thereby measurin...
no. 2 example
[0052] Example 2: Production of a mixed preparation of oil-decomposing bacterial strains with excellent soil permeability
[0053] First, a description of the permeability experiments follows.
[0054] Figure 4 It is a diagram showing microcolumns used in soil permeability experiments of microorganisms in Examples of the present invention.
[0055] As mentioned above, the dried soil was re-screened using a soil volume of 40-70 mesh, and in order to carry out the soil permeability experiment on the above-mentioned microorganisms, a plastic syringe with a small volume (for example, 3 mL) was used as Figure 4 After peeling off the filter paper (Whatman No.4), inject 1.5g of the screened soil into a 40-70mesh soil body. At this time, after the soil poured into each microcolumn was uniformly filled, the volume was kept constant.
[0056] In order to remove the influence of free-standing bacteria in the soil, after 20 minutes of wet sterilization at 121°C, put it in a dry box (...
no. 3 example
[0072] The third experimental example: Soil improvement method of oil-decomposing bacteria strain
[0073] First, osmotic regulation based on chemical changes in the cell wall of microorganisms is explained as follows.
[0074] After performing the above-mentioned step S104, as microorganisms judged to be good in oil decomposing property and used for soil permeability, Acinetobacter calcoaceticus, Klebsiella pneumoniae, Enterobacter carcinogens In the osmotic method, methyl methanesulfonate (methyl methanesulfonate; MMS, Sigma) was used to perform cell wall composition changes of the above-mentioned microorganisms (step S105 ). Furthermore, after each microorganism was cultured overnight (for example, 16 hours to 18 hours) in the M9 minimal medium, it was re-inoculated in the state of OD6000.05~0.1 in the minimal medium, and cultured to OD6000.6~0.8 ( exponential phase). The microorganisms cultivated to the index were centrifuged (8000rpm, 10min) to remove the supernatant, a...
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