CN103773713B - There is microorganism formulation of high Biofilm Colonization efficiency and preparation method thereof - Google Patents

Abstract

The invention discloses a microbial preparation with high biofilm-hanging efficiency and a preparation method thereof. Afterwards, aerobic or fermentation is carried out to obtain the bacterial agent, and the above-mentioned bacterial agent is mixed in proportion to obtain the required biological preparation; the microbial preparation includes 5-30% of the bacterial agent of Bacillus subtilis, 5-30% of the bacterial agent of Rhodopseudomonas palustris 10-40%, nitrifying bacterium 10-60%, denitrifying Brachymonas 10-40%. The preparation method of the invention can be produced on a large scale, has high efficiency and low cost, and multiple bacterial agents can be fermented and prepared at the same time, thereby shortening the time. The membrane cell density prepared by using the preparation is high, good in mass transfer, stable, long in validity, and reusable. The preparation can also increase the growth rate of the biofilm, shorten the time of biofilm formation, and improve The number of dominant microorganisms in the

Description

Microbial preparation with high biofilm-hanging efficiency and preparation method thereof

technical field

The invention relates to a microbial preparation, in particular to a microbial preparation with high biofilm formation efficiency and a preparation method thereof.

Background technique

The use of biofilm is to make wastewater flow through the biofilm grown on the surface of the fixed support, and use biological oxidation and material exchange between phases to degrade organic pollutants in wastewater. It is a method of aerobic biological treatment of wastewater. kind. Structures that use biofilm to treat wastewater include biofilters, biological turntables, and biological contact oxidation ponds. Biofilm is an ecosystem composed of highly dense aerobic bacteria, anaerobic bacteria, facultative bacteria, fungi, protozoa and algae, and the solid medium attached to it is called filter material or carrier.

The biofilm can be divided into anaerobic layer, aerobic layer, attached water layer and moving water layer from the filter material outward. The principle of the biofilm method is that the biofilm first adsorbs and attaches to the organic matter in the water layer, and the aerobic bacteria in the aerobic layer decompose it, and then enter the anaerobic layer for anaerobic decomposition, and the flowing water layer washes away the aging biofilm to grow The new biofilm reciprocates in this way to achieve the purpose of purifying sewage.

Patent CN1928081B discloses an artificial biofilm and its preparation method. Concentrated after fermentation with beneficial microbial strains. Polyvinyl alcohol, sodium alginate, sodium carboxymethyl cellulose and other film-forming raw materials are dissolved in water to make glue, and then coal-based activated carbon powder, diatomaceous earth powder, sodium acetate and single or multiple microbial cells After mixing, add it to the glue solution, and then place it in a spray device to pressurize and spray it onto the powder bed to fix it to form an artificial biofilm.

The preparation and growth of biofilm can also be a hanging film, which refers to the process of growing slowly after the microorganisms in the sewage attach. The growth time generally takes 15-20 days to complete the film hanging. There are many types of microorganisms on biofilms, including bacteria, fungi, algae, protozoa and metazoans, as well as microscopic animals visible to the naked eye. There are also differences in the types of microorganisms that make up the biofilm in the upper, middle, and lower layers of the biofilter, so the time it takes to hang the film is also different. The time of hanging film is too long, which is not conducive to the wide application and long-term recycling of biofilm.

In order to solve the problems of long film-forming time and many processes, the patent CN202829708U discloses a method for rapid film-forming of suspended filler microorganisms. It is characterized in that firstly the sludge is taken back and then aerated for 48 hours, and the aerated sludge is used as the inoculum, inoculated in the sequencing batch SBR reactor equipped with suspended filler, aerated for 6 hours, and left to settle for 6 hours; then Add nutrient substrate, aerate for 10 hours, and let it settle for 2 hours as an operation cycle, and run 2 cycles a day, during which the sludge is discharged to keep the concentration of suspended solids in the sludge at a certain concentration within the range of 2-20g/L. ; Run 7-10d to complete the hanging film.

The above-mentioned patents are mainly to improve the operation method and process. Although the speed of film formation is improved, the original film formation process is more complicated, which is not conducive to popularization and use.

Contents of the invention

The object of the invention is a biological agent and a preparation method thereof which can effectively improve the biofilm-hanging efficiency.

In order to achieve the above object, the first aspect of the present invention provides a method for preparing a biological agent with high biofilm-hanging efficiency, and the specific steps include:

In step 1, Bacillus subtilis, Nitrothorn bacteria, and Brachyclomonas denitrificans are separately cultured by aerobic aerated fermentation to obtain bacterial agents, and Rhodopseudomonas palustris is cultured by anaerobic light fermentation to obtain bacterial agents;

Step 2, mixing each bacterial agent obtained in step 1 to obtain the required biological agent, wherein, the weight ratio of each bacterial agent mixed is 5-30% of Bacillus subtilis bacterial agent, 10-30% of Rhodopseudomonas palustris bacterial agent 40%, nitrifying bacterium 10-60%, denitrifying Brachymonas 10-40%.

The mixing weight ratio of the Bacillus subtilis and the medium before the aerobic fermentation in the above step 1 is preferably 1-10:90-99, more preferably 1-3:97-99.

The temperature of aerobic aeration fermentation in the above step 1 is preferably 30-39°C, more preferably 35-38°C, most preferably 37°C, and the fermentation time is preferably 1-5 days, more preferably 1-3 days, most preferably Preferably 2 days.

The Bacillus subtilis described in the above step 1 is preferably the strain with the preservation number No. ACCC10619.

The culture medium described in the above step 1 is a mixture of at least two kinds of soybean meal powder, peptone, corn flour, sucrose, potassium dihydrogen phosphate, disodium hydrogen phosphate, ammonium sulfate, urea or sterile water.

The culture medium in the aerobic aerated fermentation culture process of Bacillus subtilis described in the above step 1 comprises: soybean meal powder 2.0-3.5%, peptone 0-1.0%, corn flour 3-4%, glucose 0-4%, disodium hydrogen phosphate 0-1%, potassium dihydrogen phosphate 0-1%, ammonium sulfate 0-2%, urea 0-1%, water 82.5-95%.

The mixing weight ratio of Rhodopseudomonas palustris and medium before anaerobic light fermentation described in the above step 1 is preferably 1-15:85-99, more preferably 2-13:90-99, more preferably 5 -10:90-95.

The temperature of anaerobic light fermentation in the above step 1 is preferably 30-38°C, more preferably 30-35°C, more preferably 32°C, and the time of anaerobic light fermentation is preferably 2-10 days, more preferably 3- 8 days, most preferably 5 days.

The Rhodopseudomonas palustris described in the above step 1 is preferably the strain with the preservation number No. ACCC10649.

The culture medium described in the above step 1 is yeast powder, sodium acetate, ammonium sulfate, potassium dihydrogen phosphate, magnesium sulfate, sodium chloride or a mixture of at least two in sterile water.

The culture medium in the anaerobic light fermentation culture process of Rhodopseudomonas palustris described in the above step 1, according to the proportion by weight, includes: yeast powder 1-2%, sodium acetate 1-5%, ammonium sulfate 1 -3%, potassium dihydrogen phosphate 0-1%, magnesium sulfate 0-1%, sodium chloride 0.5-1%, water 87-99.2%.

The mixing weight ratio of the nitrifying bacterium and the culture medium in the above step 1 is preferably 1-15:85-99, more preferably 3-12:90-99, more preferably 5-10:90-95.

The fermentation temperature of the nitrifying thorn bacteria described in the above step 1 is preferably 25-35°C, more preferably 25-30°C, most preferably 28°C, and the fermentation time is preferably 2-7 days, more preferably 2 - 5 days, most preferably 3 days.

The Nitrospina sp described in the above step 1 is preferably the strain with the preservation number No. MCCC1A00556.

The culture medium described in the above step 1 is a mixture of at least two kinds of sodium bicarbonate, sodium nitrite, sodium carbonate, potassium dihydrogen phosphate, magnesium sulfate, sodium chloride and sterile water.

The culture medium in the anaerobic light fermentation culture process of the nitrifying thorn bacteria described in the above step 1, according to the proportion by weight, includes: sodium bicarbonate 0.1-0.5%, sodium nitrite 0.1-0.5%, sodium carbonate 0.1-0.5%. 0.5%, potassium dihydrogen phosphate 0-1%, magnesium sulfate 0-1%, sodium chloride 0.5-1%, water 95.5-96.5%.

In the above step 1, the mixing weight ratio of the denitrifying Brachymonas to the culture medium is preferably 1-8:92-99, more preferably 1-5:95-99, more preferably 1-2:98-99 .

The temperature of the fermentation described in the above step 1 is preferably 25-35°C, more preferably 25-30°C, most preferably 28°C, and the fermentation time is preferably 1-8 days, more preferably 1-8 days 5 days, more preferably 2-4 days, most preferably 3 days.

The denitrifying Brachymonas denitrificans described in the above step 1 is preferably a strain with a preservation number of No. CGMCC1.7100.

The culture medium described in the above step 1 is sodium acetate, potassium nitrate, potassium dihydrogen phosphate, magnesium chloride, calcium chloride or a mixture of at least two in sterile water.

The culture medium in the denitrifying Brachymonas aerobic aerated fermentation culture process described in the above step 1 comprises: sodium acetate 0.01-0.05%, potassium nitrate 0.01-0.05%, dihydrogen phosphate Potassium 0.001-0.002%, magnesium chloride 0.001-0.002%, calcium chloride 0.001-0.002%, water 99.894-99.97%.

The second aspect of the present invention is to provide a microbial preparation prepared by the above method, including microbial components, based on the total amount of microbial components, including the following strains by weight percentage:

Bacillus subtilis bacterial agent is preferably 5-30%, more preferably 10-25%, more preferably 10-20%;

Rhodopseudomonas palustris bacteria agent is preferably 10-40%, more preferably 10-30%, more preferably 20-30%;

The nitrifying bacterium bacterium agent is preferably 10-60%, more preferably 20-50%, more preferably 30-40%;

10-40% of the denitrifying Brachymonas bacterial agent, more preferably 20-40%, more preferably 20-30%.

The preparation method of the microbial preparation with high biofilm-hanging efficiency described in the present invention can realize large-scale production, the process is simple, the efficiency is high and the cost is low, and the fermentation preparation of various microbial preparations can be carried out simultaneously, thereby greatly shortening the microbial preparation. Preparation time, improve production efficiency.

The cell density in the film obtained by using the microbial preparation of the present invention is high, good in mass transfer, stable, long in validity, reusable, and can increase the growth rate of the biofilm, shorten the film-hanging time of the biofilm, and improve the biological efficiency of the biofilm. The advantage of the number of microorganisms in the membrane is the number of aerobic and anaerobic predominance.

Description of drawings

Fig. 1 is the graph of COD removal situation changing with time during the aerobic culture of contrasting 0% inoculum size in the embodiment of the present invention 3;

Fig. 2 is the graph of COD removal situation changing with time during the aerobic culture of contrasting 0.05% inoculum size in the embodiment of the present invention 3;

Fig. 3 is the figure of COD removal over time during the aerobic culture of the control 0.075% inoculum size in Example 3 of the present invention;

Fig. 4 is the figure of COD removal over time during the aerobic culture of the control 0.1% inoculum size in Example 3 of the present invention;

Fig. 5 is the graph of COD removal situation changing with time during the anaerobic culture of contrasting 0% inoculum size in the embodiment of the present invention 3;

Fig. 6 is the graph of COD removal situation changing with time during the anaerobic culture of contrasting 0.05% inoculum size in the embodiment of the present invention 3;

Fig. 7 is a time-varying graph of COD removal during anaerobic culture with an inoculum size of 0.075% in Example 3 of the present invention.

specific embodiment

Biofilm method has unique advantages in sewage treatment, especially aquaculture wastewater treatment, because of its advantages of less sludge generation, convenient operation and management, and low treatment cost. By adding the same genus or similar microorganisms, the growth efficiency of the biofilm will be improved to a certain extent, and the time for its film formation can also be shortened. Therefore, the use of appropriate microbial agents can effectively increase the growth rate of biofilms and increase the number of necessary aerobic and anaerobic dominant microorganisms in biofilms, thereby improving the quality of biofilms and reducing the time for hanging films.

The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments, but not as a limitation of the present invention.

Example 1

The Bacillus subtilis described in this example is a strain with the preservation number No. ACCC10619.

The said Rhodopseudomonas palustris is a strain with the preservation number No. ACCC10649.

The Nitrospina sp is a strain with the preservation number No. MCCC1A00556.

The denitrifying Brachymonas denitrificans (Brachymonas denitrificans) is a strain with a preservation number of No. CGMCC1.7100.

The preparation can have the method for the microbial preparation of high biofilm hanging film efficiency, and its specific steps are as follows:

Step 1, the preparation of Bacillus subtilis bacterial agent.

Before aerobic fermentation, it is prepared from the following raw materials in weight percentage: 1% of Bacillus subtilis and 99% of culture medium, and is subjected to aerobic aerobic fermentation at 37°C for 2 days. After fermentation, the fermented liquid is ready for use.

The proportion by weight of the medium in step 1 is: 2.0% of soybean meal powder, 1.0% of peptone, 3.0% of corn flour, 0.75% of disodium hydrogen phosphate, 0.2% of potassium dihydrogen phosphate, and 93.05% of water.

Step 2, preparation of Rhodopseudomonas palustris bacterial agent.

Before anaerobic light fermentation, it is prepared from the following raw materials in weight percentage: 5% of Rhodopseudomonas palustris and 95% of medium, and is subjected to anaerobic light fermentation at 32° C. for 5 days. After fermentation, the fermented liquid is ready for use.

The proportion by weight of the medium in step 2 is: yeast powder 1.0%, sodium acetate 5.0%, ammonium sulfate 3.0%, sodium chloride 0.5%, water 90.5%.

Step 3, preparation of the nitrophilic bacteria agent.

Before the aerobic fermentation, it is prepared from the following raw materials in weight percentage: 10% of Nitrophilus and 90% of culture medium, and is subjected to aerobic aerobic fermentation at 28°C for 3 days. After fermentation, the fermented liquid is ready for use.

The proportion by weight of the medium in step 3 is: 0.5% sodium bicarbonate, 0.1% sodium nitrite, 0.5% sodium carbonate, 0.5% magnesium sulfate, 0.5% sodium chloride, and 97.9% water.

Step 4, preparation of denitrifying Brachymonas bacterial agent.

Before aerobic fermentation, it is prepared from the following raw materials in weight percentage: 1% of Brachymonas denitrificans, 99% of culture medium, and is subjected to aerobic aerobic fermentation at 28° C. for 3 days. After fermentation, the fermented liquid is ready for use.

The proportion by weight of the culture medium in step 4 is: 0.05% sodium acetate, 0.05% potassium nitrate, 0.002% potassium dihydrogen phosphate, 0.001% magnesium chloride, 0.001% calcium chloride, and 99.896% water.

Step 5, preparation of biological preparations. Mix Bacillus subtilis bacterial agent, Rhodopseudomonas palustris bacterial agent, nitrifying bacterium bacterial agent and denitrifying Brachymonas bacterial agent, and its percentage by weight is: Bacillus subtilis bacterial agent 20%, swamp red false 30% of monocyst bacteria, 30% of nitrifying sting bacteria, 20% of denitrifying Brachymonas bacteria.

After mixing, the desired microbial preparation is obtained.

Example 2

The Bacillus subtilis described in this example is a strain with the preservation number No. ACCC10619.

The said Rhodopseudomonas palustris is a strain with the preservation number No. ACCC10649.

The Nitrospina sp is a strain with the preservation number No. MCCC1A00556.

The denitrifying Brachymonas denitrificans (Brachymonas denitrificans) is a strain with a preservation number of No. CGMCC1.7100.

The method for preparing the microbial preparation with high biofilm hanging film efficiency, its specific steps are as follows:

Step 1, the preparation of Bacillus subtilis bacterial agent.

Before aerobic fermentation, it is prepared from the following raw materials in weight percentage: 3% of Bacillus subtilis and 97% of medium, and is subjected to aerobic aerobic fermentation at 37°C for 2 days. After fermentation, the fermented liquid is ready for use.

The proportion by weight of the medium in step 1 is: soybean meal 3.0%, corn flour 4.0%, disodium hydrogen phosphate 0.75%, potassium dihydrogen phosphate 0.2%, ammonium sulfate 0.5%, water 91.55%.

Step 2, preparation of Rhodopseudomonas palustris bacterial agent.

Before the anaerobic photo-fermentation, it is prepared from the following raw materials in weight percentage: 10% of Rhodopseudomonas palustris and 90% of the culture medium, and is subjected to anaerobic photo-fermentation at 32° C. for 5 days. After fermentation, the fermented liquid is ready for use.

The proportion by weight of the medium in step 2 is: 2.0% yeast powder, 5.0% sodium acetate, 0.05% potassium dihydrogen phosphate, 0.05% magnesium sulfate, 0.5% sodium chloride, and 92.4% water.

Step 3, preparation of the nitrophilic bacteria agent.

Before the aerobic fermentation, it is prepared from the following raw materials in weight percentage: 10% of Nitrophilus and 90% of culture medium, and is subjected to aerobic aerobic fermentation at 28°C for 3 days. After fermentation, the fermented liquid is ready for use.

The proportion by weight of the medium in step 3 is: 0.5% sodium bicarbonate, 0.5% sodium nitrite, 0.5% potassium dihydrogen phosphate, 0.5% magnesium sulfate, 0.5% sodium chloride, and 97.5% water.

Step 4, preparation of denitrifying Brachymonas bacterial agent.

Before aerobic fermentation, it is prepared from the following raw materials in weight percentage: 2% of Brachymonas denitrificans, 98% of culture medium, and is subjected to aerobic aerobic fermentation at 28° C. for 3 days. After fermentation, the fermented liquid is ready for use.

The proportion by weight of the medium in step 4 is: 0.05% sodium acetate, 0.03% potassium nitrate, 0.002% potassium dihydrogen phosphate, 0.002% calcium chloride, and 99.916% water.

Step 5, preparation of biological preparations.

Mix the Bacillus subtilis bacteria agent, Rhodopseudomonas palustris bacteria agent, nitrifying bacterium bacteria agent and denitrifying Brachymonas bacteria agent, and its percentage by weight is: Bacillus subtilis bacteria agent 10%, swamp red fake 30% of monocyst bacteria agent, 30% of nitrifying bacterium bacteria agent, and 30% of denitrifying Brachymonas bacteria agent.

Example 3

Experiments on the effect of the microbial preparation of the present invention on the efficiency of biofilm formation in anaerobic and aerobic environments.

The purpose of the experiment: Inoculate the complex bacteria into the anaerobic-aerobic combination reactor, and add the pig farm wastewater that has been pretreated by sedimentation or coagulation, and conduct the film-hanging test by inoculating the complex bacteria. Experiments were carried out according to the difference before and after the addition to investigate its effect on shortening the film-forming time and improving the removal of pollutants.

Experimental sewage: The livestock and poultry breeding wastewater used in this experiment is mainly characterized by turbid water quality and certain impurities, dark brown in color and certain odor. In this experiment, COD, the main water quality indicator of sewage, is tested, and its range is about 3000mg/L.

Experimental device: The experimental device used in this experiment is a cuboid made of plexiglass, with a size of 255mm×65mm×225mm. The reactor includes three independent compartments, and each compartment is a small reactor. Packing is installed in each cell for cultivating microorganisms. Waste water is installed in the reactor, the microorganisms are cultivated and domesticated, and the simulated pig farm waste water is injected, and the COD in the waste water is removed by the microorganisms. Each compartment is aerated or sealed according to the test requirements (aerobic or anaerobic).

Experimental method: In this experiment, the cultivation and domestication of biofilms are carried out on the filler, so the filler is first fixed in each small unit of the reactor, and hung evenly and vertically in each small cell by tying. . Aeration devices are then added to each cell to provide the oxygen needed for microbial growth. In this experiment, two groups of microorganisms were tested under anaerobic and aerobic conditions.

The first stage is to cultivate microorganisms under aerobic conditions, and add artificial simulated aquaculture wastewater into the two cells respectively (artificial aquaculture wastewater is prepared from sucrose, ammonium chloride and potassium dihydrogen phosphate plus tap water to make C:N:P 100:5:1, and then add 100mL of aquaculture wastewater to keep its COD at about 1000mg/L) Batch method is used to cultivate microorganisms (the metering pump does not enter water, but continuous aeration), according to 0.05%, 0.075% and 0.1% three inoculum ratios to inoculate the strains, and the aerobic hanging film is cultivated for about a week. The specific method is to add simulated aquaculture wastewater, continuously aerate and inoculate the filler, keep the influent COD at about 1000mg/L, carry out intermittent method to cultivate microorganisms, test the COD value of influent and effluent water every other day, and determine the COD removal rate based on the judgment basis after one week The end and time point of the aerobic film formation phase.

The second stage is to cultivate the biofilm under anaerobic conditions. The first compartment of the reactor stops aeration to meet anaerobic conditions, and the second compartment is aerated. Adopt the intermittent method to cultivate anaerobic microorganisms (the metering pump does not enter the water), add artificial breeding wastewater (the concentration is the same as above, about 1000mg/L), and insert the bacteria according to the three inoculation ratios of 0.05%, 0.075% and 0.1%, and cultivate approximately The anaerobic film formation 10d ends. Specifically, when the aerobic microorganisms are matured, anaerobic microorganisms are cultivated on the basis of aerobic microorganisms, the aeration equipment is removed, the aeration is stopped, the concentration of the distribution water is increased to greater than 1000mg/L, and the COD of the influent and effluent is tested. Oxygen bacteria culture is over.

The mixing weight ratio and the inoculum amount of the composite strains used in the experiment are shown in Table 1-2.

The aerobic and anaerobic film-hanging days of the microbial preparation of the present invention are shown in Table 3.

Table 1, the list of inoculation weight ratios of multi-strains used in experiments

strain name Bacteria content Proportion Rhodopseudomonas palustris 2 billion/ml 30% Nitrobacter 0.1 million/ml 30% Brachymonas denitrificans 0.1 million/ml 30% Bacillus subtilis 5 billion/ml 10%

Table 2, the list of the inoculum amount of the composite bacteria solution

Table 3, the aerobic and anaerobic film-hanging days comparison table of the microbial preparation of the present invention

The results of COD removal during aerobic culture and COD removal during anaerobic culture are shown in Figure 1-7.

Analysis of results:

(1) The trend of COD removal over time during aerobic culture. Figure 1-4 is a time-dependent graph of COD removal during aerobic cultivation. As shown in Figure 1-4, as the influent COD concentration is maintained at about 1000 mg/L, the effluent COD value has a significant downward trend, and the COD removal rate gradually increase. Starting from the influent COD of 1052mg/L, the removal rate gradually increased from 9.89% over time, and reached the maximum removal rate of 96.06% after one week when the influent COD was 1022.5mg/L, indicating that the aerobic stage was well cultivated Oxygen microorganisms meet the requirements of water purification. During this period, the color of the biofilm on the filler became darker, from white filaments gradually to yellow and then to brown, the thickness of the film increased significantly, and the microbial culture was successful.

And comparing Figures 1-4, it can be seen that with the increase of the inoculum amount of the microbial preparation of the present invention, the COD removal efficiency is significantly improved, and the microbial preparation of the present invention can make the COD removal efficiency significantly improved after four days.

(2) Comparing Figures 5-7, with the increase of the inoculum amount of the microbial preparation of the present invention, the COD removal efficiency also increased significantly from the sixth day; within the first 6 days, the facultative bacteria grown under aerobic conditions entered the anaerobic environment Afterwards, an adaptation process is required, and the change trend of COD removal efficiency is not obvious, but even so, even a small amount of oxygen can enhance the activity of facultative bacteria in this process (as shown in Figure 6 and Figure 7), so acidification Bacteria multiply from top to bottom. Different inoculum amounts had an effect on COD consumption and biofilm formation under anaerobic conditions.

(3) From Tables 2 and 3, it can be seen that the inoculation amount of different strains is set under aerobic and anaerobic conditions, and it can be seen that there is a difference in the formation time of the final biofilm. Under aerobic conditions, when the inoculum amount is 0.075 %, the biofilm formation time was shortened by 1 day compared with the control group, and the cost was lower than that of the 0.1% inoculum group; under anaerobic conditions, the 0.1% inoculum amount had the best effect. That is, the inoculum amount of 0.075% and 0.1% under aerobic conditions and the inoculum amount of 0.1% under anaerobic conditions all help to reduce the time of film formation and reduce the cost of treatment.

To sum up, in the embodiment of the present invention, in the aerobic and anaerobic two groups of experiments, the strains were added, and the bacterial content of the compound bacterial liquid in the added strains was 2 billion/ml, successfully realizing the The cultivation of aerobic and anaerobic biofilms can effectively improve the efficiency of biofilms.

The specific embodiments of the present invention have been described in detail above, but they are only examples, and the present invention is not limited to the specific embodiments described above. For those skilled in the art, any equivalent modifications and substitutions to this practice are also within the scope of the present invention. Therefore, equivalent changes and modifications made without departing from the spirit and scope of the present invention shall fall within the scope of the present invention.

Claims (9)

1. the microorganism formulation application in terms of improving Biofilm Colonization efficiency, it is characterised in that described microorganism formulation Preparation method include:

Step 1, bacillus subtilis, nitrification thorn bacterium, denitrification brachyplast Zymomonas mobilis individually carry out aerobic aerobic fementation cultivation Obtaining microbial inoculum, Rhodopseudomonas palustris carries out anaerobism illumination fermentation culture and obtains microbial inoculum；

Step 2, each microbial inoculum step 1 obtained mixes, and obtains required biological preparation, wherein, the part by weight of each microbial inoculum mixing For bacillus subtilis microbial agent 5-30%, Rhodopseudomonas palustris microbial inoculum 10-40%, nitrification thorn bacteria agent 10-60%, denitrification Brachyplast Zymomonas mobilis microbial inoculum 10-40%.

Application the most according to claim 1, it is characterised in that the aerobic aerobic fementation of bacillus subtilis described in step 1 Culture medium in incubation, according to weight percent proportioning, including: bean cake powder 2.0-3.5%, peptone 0-1.0%, Semen Maydis powder 3-4%, glucose 0-4%, disodium hydrogen phosphate 0-1%, potassium dihydrogen phosphate 0-1%, ammonium sulfate 0-2%, carbamide 0-1%, water 82.5-95%.

Application the most according to claim 2, it is characterised in that the bacillus subtilis described in step 1 mixes with culture medium The weight ratio closed is 1-10:90-99, and the temperature of aerobic aerobic fementation is 30-39 DEG C, and the time is 1-3 days.

Application the most according to claim 1, it is characterised in that the Rhodopseudomonas palustris anaerobism illumination described in step 1 Culture medium during fermentation culture, according to weight percent proportioning, including: yeast powder 2%, sodium acetate 5%, ammonium sulfate 3%, phosphorus Acid dihydride potassium 1%, magnesium sulfate 1%, sodium chloride 1%, water 87%.

Application the most according to claim 4, it is characterised in that the Rhodopseudomonas palustris described in step 1 and culture medium The weight ratio of mixing is 1-15:85-99, and the temperature of anaerobism illumination fermentation is 30-38 DEG C, and the time is 2-10 days.

Application the most according to claim 1, it is characterised in that the nitrification thorn bacterium aerobic illumination fermentation training described in step 1 Culture medium during Yanging, according to weight percent proportioning, including: sodium bicarbonate 0.1-0.5%, sodium nitrite 0.1-0.5%, carbon Acid sodium 0.1-0.5%, potassium dihydrogen phosphate 0-1%, magnesium sulfate 0-1%, sodium chloride 0.5-1%, water 95.5-96.5%.

Application the most according to claim 6, it is characterised in that the nitrification thorn bacterium described in step 1 mixes with culture medium Weight ratio is 3-12:90-99, and the temperature of aerobic aerobic fementation is 25-35 DEG C, and the time is 2-7 days.

Application the most according to claim 1, it is characterised in that the denitrification brachyplast Zymomonas mobilis described in step 1 is aerobic logical Culture medium during gas fermentation culture is according to weight percent proportioning, including sodium acetate 0.01-0.05%, potassium nitrate 0.01- 0.05%, potassium dihydrogen phosphate 0.001-0.002%, magnesium chloride 0.001-0.002%, calcium chloride 0.001-0.002%, water 99.894-99.97%.

Application the most according to claim 8, it is characterised in that the denitrification brachyplast Zymomonas mobilis described in step 1 and cultivation The weight ratio of base mixing is 1-8:92-99, and the temperature of aerobic aerobic fementation is 25-35 DEG C, and the time is 1-8 days.