CN115287208A - Method for producing beneficial microbial agent by using pig raising wastewater while removing environmental pollutants in pig raising wastewater - Google Patents
Method for producing beneficial microbial agent by using pig raising wastewater while removing environmental pollutants in pig raising wastewater Download PDFInfo
- Publication number
- CN115287208A CN115287208A CN202210108012.0A CN202210108012A CN115287208A CN 115287208 A CN115287208 A CN 115287208A CN 202210108012 A CN202210108012 A CN 202210108012A CN 115287208 A CN115287208 A CN 115287208A
- Authority
- CN
- China
- Prior art keywords
- fermentation
- liquid
- pig
- wastewater
- raising wastewater
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/22—Bacillus
Abstract
The invention discloses a method for producing a beneficial microbial agent by utilizing pig raising wastewater while removing environmental pollutants in the pig raising wastewater. The invention utilizes single bacteria and composite beneficial microorganism strains, takes the pig-raising wastewater as a basic culture medium, adopts a step-by-step amplification culture process, and adjusts the carbon-nitrogen ratio by adding brown sugarThe beneficial microorganism fermentation production is carried out, on one hand, the production cost of the microbial fertilizer can be greatly reduced, the high-density growth of thalli in the fermentation liquor is realized, and the viable bacteria content in the prepared beneficial microorganism bacterial powder is 7 multiplied by 10 10 ‑1×10 11 cfu/g, on the other hand, the pollution problem of the pig raising wastewater can be reduced through biodegradation, the pig raising wastewater is changed into valuable, the residual value is realized, a large amount of resources for treating environmental pollution can be saved, and the method has very important significance for promoting the utilization of harmless resources of environmental pollutants.
Description
Technical Field
The invention relates to a method for producing a beneficial microbial agent by utilizing pig raising wastewater, in particular to a resource utilization method of pig raising wastewater, which utilizes beneficial microorganisms to remove environmental pollutants in the pig raising wastewater and simultaneously obtain beneficial microbial thalli. The invention belongs to the technical field of microorganisms.
Background
China is one of the most important pork consumption export countries and pig breeding countries in the world at present. A large amount of live pigs are bred to bring extremely serious burden to the environment, and most live pig farms in China adopt a flushing mode to clean pig houses, so that the main body of the pig farm wastewater contains live pig urine, excrement, other sewage and the like, and the pig farm wastewater is mixed with solid and liquid; the content of ammoniacal nitrogen and phosphorus is high; the chemical concentration of suspended matters and organic matters contained in the fertilizer is high; if a large amount of untreated pig raising wastewater is discharged into lakes and slow flows, eutrophication of water is easily caused, water sources are polluted, and the discharge of the untreated pig raising wastewater into farmlands causes the discharge of the pig raising wastewater to exceed the limit of the self-purification capacity of soil, harmful substances such as malodorous substances and nitrite and the enrichment of trace elements such as phosphorus, copper and zinc are generated, so that the soil components and properties are changed, and even after the pig raising wastewater is subjected to a conventional sewage treatment process, the concentration of pollutants such as nitrogen and phosphorus in the discharged water can still keep a high level, and the quality of the water environment of rivers and lakes in China can also be seriously threatened, so that the pollution problem of the pig raising wastewater to the environment is gradually revealed and tends to spread towards a worsening trend. However, although the pig raising wastewater exerts a great pressure on the environment, the pig raising wastewater contains a large amount of organic matters, and nutrient elements such as nitrogen, phosphorus, potassium and the like, and is an important resource, wherein a considerable proportion of the nutrient elements can be utilized, so that the resource utilization potential is great, and an effective mode is urgently needed for utilization.
Disclosure of Invention
The invention aims to provide a method for producing a beneficial microbial agent by utilizing pig raising wastewater while removing environmental pollutants in the pig raising wastewater. The invention utilizes single bacteria and composite beneficial microorganism strains to carry out beneficial microorganism fermentation production by taking the pig-raising wastewater as a basic culture medium, on one hand, the production cost of the microorganism bacterial manure can be greatly reduced, on the other hand, the pollution problem can be reduced through biodegradation, the pig-raising wastewater is changed into valuable, the residual value is realized, a large amount of resources for treating environmental pollution can be saved, and the invention has very important significance for promoting the harmless resource utilization of environmental pollutants.
In order to achieve the purpose, the invention adopts the following technical means:
the invention discloses a method for producing a beneficial microbial agent by using pig raising wastewater while removing environmental pollutants in the pig raising wastewater, which comprises the following steps:
step one, strain activation: inoculating beneficial microorganisms preserved at-80 ℃ into a liquid culture medium, inoculating the preserved strains in a centrifugal tube into a 250mL triangular flask filled with 100mL of the liquid culture medium by using an inoculating loop under the aseptic operation condition, and performing shake culture at 30 ℃ for 24h at 180r/min for activation to obtain activated bacteria liquid for later use;
wherein the liquid culture medium comprises the following components: according to the mass percentage, the liquid culture medium contains 0.3 to 0.4 percent of beef extract, 1 to 1.2 percent of peptone, 0.5 percent of sodium chloride and the balance of water;
step two, seed liquid culture: inoculating the liquid activated bacterium liquid prepared in the step one into a 2L triangular flask filled with 500mL of seed culture medium according to the inoculation amount of 1-3 vol%, and performing shake culture at 34 ℃ for 24h at 180r/min for seed liquid culture to obtain seed liquid for later use;
the seed culture medium comprises the following components: according to the mass percentage, the seed culture medium contains 1 to 1.5 percent of brown sugar, and the rest is pig raising wastewater;
step three, first-stage seed fermentation tank culture: inoculating the seed solution cultured in the second step into a 50L fermentation tank containing 50L fermentation medium, wherein the inoculation amount is 1% of the total volume of the fermentation medium, culturing at 35-38 deg.C for 24-36h at 100-150r/min, and introducing sterile air continuously until the viable count reaches 2 × 10 10 -4×10 10 Obtaining seed fermentation liquor after cfu/mL for later use;
the fermentation medium comprises the following components: according to the mass percentage, the fermentation medium contains 1 to 1.5 percent of brown sugar, and the rest is pig-raising wastewater;
step four, secondary fermentation tank amplification culture: inoculating the seed fermentation liquid cultured in the third step into a 500L fermentation tank containing 500L fermentation medium, wherein the inoculation amount is 5% -10% of the volume of the fermentation tank, culturing at 35-38 ℃ at 100-150r/min for 24h, continuously introducing sterile air during the period, and when the viable count reaches 2 x 10 10 -4×10 10 Obtaining seed fermentation liquor for later use after cfu/mL;
the components of the fermentation medium are consistent with those of the fermentation medium in the third step;
step five, third-stage fermentation tank expanded culture: inoculating the fermentation liquid cultured in the fourth step into a three-stage fermentation tank filled with 3000L of fermentation medium, wherein the inoculation amount is 5% -10% of the volume of the fermentation tank, culturing at 37-39 deg.C and 60-80r/min for 24-48 h, continuously introducing sterile air during the period, and when the viable count reaches 4 × 10 10 Stopping culturing after cfu/mL or more, and discharging to obtainFermentation liquor;
the components of the fermentation medium are consistent with those of the fermentation medium in the third step;
step six, bacterial liquid flocculation: adding a bioflocculant into the obtained fermentation liquor, stirring and standing to obtain a settled thallus flocculation liquid;
seventhly, plate and frame filter pressing: uniformly mixing the settled thallus flocculation liquid obtained in the sixth step at 150r/min, and filtering by using a plate-and-frame filter press to obtain flocculated wet bacteria and fermentation waste liquid;
step eight, drying the raw powder: putting the wet thalli obtained after filter pressing in the step seven into a vertical boiling dryer for drying, wherein the drying temperature is 40 ℃, and the drying time is 6h;
ninth, beneficial microorganism powder preparation: mixing the beneficial microorganism raw powder obtained in the step eight with anhydrous calcium carbonate, wherein the mixing mass ratio is 1:15 to 20, stirring in a stirrer for 0.5 to 2 hours, crushing and sieving by a 100-mesh sieve to obtain the beneficial microbial powder.
Wherein, preferably, the beneficial microorganisms comprise: LZP03 (Bacillus megaterium), LZP (Bacillus pumilus), WB (Bacillus velezensis) and SC (Bacillus subtilis), wherein the beneficial microorganisms are all preserved in China center for type culture Collection with the preservation numbers of CCTCC NO: m2018599, CCTCC NO: m2018598, CCTCC NO: m20211547, CCTCC NO: m20211546.
Relevant experiments prove that LZP (Bacillus megaterium) and LZP (Bacillus pumilus) as a rice rhizosphere growth-promoting bacterium can effectively promote the growth and development of rice seedlings and improve the nutrient absorption capacity of the rice seedlings, WB (Bacillus velezensis) can obviously inhibit the growth of crop pathogenic bacteria Fusarium oxysporum, improve the disease resistance of crops and promote the yield and income of the crops, and SC (Bacillus subtilis) has good capability of preventing and treating watermelon fusarium wilt and is a high-efficiency biocontrol bacterium.
Wherein, preferably, the beneficial microorganism consists of LZP (Bacillus megaterium), LZP (Bacillus pumilus), WB (Bacillus velezensis), SC (Bacillus subtilis) in a volume ratio of 1.
Preferably, the bacterial liquid in the sixth step is flocculated, and the specific mode is that the bioflocculant is added into the obtained fermentation liquor, the fermentation liquor is quickly stirred at 200r/min for 10min, the fermentation liquor is slowly stirred at 30r/min for 30min, and the fermentation liquor is kept stand for 60min to obtain settled thallus flocculation liquid.
Further, the invention provides beneficial microbial powder prepared by the method.
Furthermore, the invention provides the application of the beneficial microbial powder in the aspects of crop growth promotion and crop biocontrol.
Compared with the prior art, the invention has the beneficial effects that:
1. the invention provides a method for producing a beneficial microbial agent by using pig wastewater while removing environmental pollutants in the pig wastewater, which takes beneficial microorganisms as strains, each strain is screened at the rhizosphere of crops, has good plant growth promoting and biocontrol functions, and is verified to be a non-pathogenic strain; the pig raising wastewater fermentation culture medium has low cost; the method adopts a step-by-step amplification culture process and adjusts the carbon-nitrogen ratio by adding brown sugar to realize the high-density growth of thalli in the fermentation liquor, and the content of the prepared beneficial microbial powder is 7 multiplied by 10 10 -1×10 11 cfu/g。
2. According to the verification, in the fermentation process, whether beneficial single-bacterium fermentation or composite-bacterium fermentation is carried out, the types of various pollutants in the pig raising wastewater are obviously reduced, and COD (chemical oxygen demand), ammoniacal nitrogen, organic matters and phosphorus content in the fermentation waste liquid after thallus removal have obvious removal rate. Therefore, the method reduces the pollution sources such as ammoniacal nitrogen, organic matters, phosphorus and the like which threaten the environmental safety in the pig raising wastewater on one hand, and can collect microbial preparations for various aspects such as plant growth promotion, crop biocontrol, pollutant degradation and the like through a large amount of microbial thalli cultured by the pig raising wastewater on the other hand, achieves two purposes by one action, and the higher cost of a culture medium of the microbes in the culture process is one of the main reasons for keeping the price of the microbial preparation high, while the beneficial microbial strains cultured by the pig raising wastewater are close to the traditional culture medium in viable count, but the cost is greatly reduced, thereby having important significance for realizing green agriculture by reducing the use of agricultural chemicals and chemical fertilizers by popularizing the microbial preparation.
Drawings
FIG. 1 shows the change of biomass of the single bacterial agent and the compound bacterial agent in the optimized pig raising wastewater along with time;
wherein, A: LZP02; b: LZP03; c: SC; d: WB; e: LZP 03-LZP-WB-SC complex bacteria;
FIG. 2 is a graph showing the results of total nitrogen removal by each strain in the fermentation process of wastewater from swine;
FIG. 3 is a graph showing the results of the total carbon removal rate of each strain in the fermentation process of the swine wastewater;
FIG. 4 is a graph showing the results of the total organic carbon removal rate of each strain in the fermentation process of the swine wastewater;
FIG. 5 is a graph showing the trend of pH changes with time of each strain in the fermentation process of swine wastewater;
FIG. 6 is a graph showing the results of COD removal by each strain in swine wastewater;
FIG. 7 is a graph showing the results of removing ammoniacal nitrogen from swine wastewater by each strain;
FIG. 8 is a graph showing the results of the removal rate of phosphorus content in wastewater from swine production by each strain;
FIG. 9 is a comparison of the trend and the reduction rate of total nitrogen, total carbon and total organic carbon in the supernatant of the complex microbial inoculum fermentation broth
FIG. 10 shows the pH variation trend of the complex microbial inoculum along with fermentation time in the optimized culture medium
FIG. 11 is a graph showing the time-dependent trend of COD removal from swine wastewater in the complex microbial inoculum fermentation broth
FIG. 12 shows the removal rate of ammoniacal nitrogen and phosphorus in swine wastewater by the composite microbial inoculum
FIG. 13 is a flow chart of the production process;
FIG. 14 is a comparison of Chinese cabbage before and after use.
Strain preservation information:
the Bacillus pumilus LZP is named as Bacillus pumilus LZP02, is classified and named as Bacillus pumilus LZP02 and is preserved in China Center for Type Culture Collection (CCTCC) NO. M2018598, and the preservation time is 2018, 9 and 6 days.
The Bacillus megatherium LZP is named as Bacillus megatherium LZP03, is classified and named as Bacillus megatherium LZP03 respectively, is preserved in China center for type culture collection, and is addressed to the university of Wuhan, china, with the strain preservation number of CCTCC NO. M2018599 and the preservation time of 2018, 9 months and 6 days.
The Bacillus subtilis strain SC is named as Bacillus subtilis SC, classified and named as Bacillus subtilis SC, is preserved in China center for type culture Collection, and is addressed to Wuhan university, wuhan, china, and the strain preservation number is CCTCC NO: m20211546 with a preservation time of 2021, 12 and 6 days.
The Bacillus bailii strain WB is named as Bacillus velezensis WB, is classified and named as Bacillus velezensis WB, is preserved in China center for type culture collection, and is addressed to Wuhan university, and the strain preservation number is CCTCC NO: m20211547 with a preservation time of 2021, 12 and 6 days.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. It is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments, and all other embodiments obtained by those skilled in the art without any inventive work are within the scope of the present invention.
Example 1 comparison of growth of Single microbial Agents and Complex microbial Agents in pig wastewater
1 materials and methods
1.1 sources of test strains
The strains used in the test are different types of pure culture beneficial microorganisms obtained by separating and screening the subject components, and researches show that all strains are high-efficiency strains with better industrial application potential in the aspects of plant growth promotion, biological prevention, pollutant prevention and the like, all strains can rapidly grow in the pig raising wastewater and have better removal effect on ammonia nitrogen, phosphorus, organic matters and the like, the preservation place is a refrigerator at minus 80 ℃, all strains are classified and identified by strain morphology and 16SrDNA, and the identification list is shown in Table 1.
TABLE 1 test strains
1.2 physicochemical Properties of wastewater from pig raising
The swine wastewater used in the test was obtained from a certain pig farm with a flush Ha Ershi fural base region, and the basic characteristics of the collected swine wastewater are shown in table 2.
TABLE 2 physical and chemical Properties of the original wastewater from pig raising
1.3 Medium
Beef extract peptone medium: 10.00g of peptone, 3.00g of beef extract, 5.00g of NaCl, 20.00g of agar, and adding no agar into a liquid culture medium, wherein the volume of distilled water is up to 1000.00mL.
The pig raising wastewater culture medium after the carbon-nitrogen ratio is optimized: 10g of brown sugar and the volume of the pig raising wastewater is up to 1000.00mL.
1.4 methods of investigation
(1) Single strain fermentation culture
Beneficial microorganism strains LZP, LZP, WB and SC preserved at-80 ℃ are respectively inoculated into beef extract peptone solid culture media and cultured for 24h at 30 ℃ for activation. Inoculating 2-3 rings of activated strains into a 250mL triangular flask filled with 100mL beef extract peptone liquid culture medium under the aseptic operation condition by using an inoculating ring, carrying out shaking culture at 30 ℃ and 120r/min, then taking each strain cultured to the same concentration (OD 600= 1.0) as a seed solution, then respectively inoculating activated bacteria liquid of LZP03, LZP, WB and SC into 300mL pig raising wastewater culture medium according to the inoculation amount of 1vol%, then carrying out shake flask culture at 30 ℃ and 120r/min, sampling samples every 24h, and determining the growth condition of a single bacterium agent in the optimized pig raising wastewater culture medium along with time;
(2) Fermentation culture of composite strain
Beneficial microorganism strains LZP, LZP, WB and SC which are preserved at-80 ℃ are respectively inoculated into a beef extract peptone solid medium and cultured for 24h at 30 ℃ for activation. The strains after 2-3 rings activation are inoculated in a 250mL triangular flask containing 100mL beef extract peptone liquid medium under aseptic operation conditions by using an inoculating ring, shaking culture is carried out at 30 ℃ and 120r/min, then each strain cultured to the same concentration (OD 600= 1.0) is used as seed liquid, activated bacteria liquid of LZP, LZP, WB and SC are mixed according to the volume ratio of 1.
2. Results
In order to determine the change of the biomass of the single microbial inoculum and the compound microbial inoculum in the culture medium along with the time, the activated strains are inoculated in the optimized culture medium to carry out viable count determination by a gradient dilution plating method.
As shown in FIG. 1A, LZP cultured in the optimized pig-raising wastewater culture medium for 72h reaches the maximum viable count of 4.07X 10 10 cfu/mL. As shown in FIG. 1B, LZP optimized in the swine wastewater culture medium reaches the maximum viable cell count of 4.26X 10 in 96h 10 cfu/mL. As can be seen from FIG. 1C, SC strain was cultured in pig-raising wastewater culture medium with optimized carbon-nitrogen ratio for 48h to reach a maximum viable count of 4.16X 10 10 Viable count of cfu/mL. As can be seen from FIG. 1D, the WB strain reached a maximum biomass of 4.64X 10 after 48h in the original medium 10 cfu/mL. From FIG. 1E, it can be seen that the biomass of the composite microbial inoculum growing in the pig wastewater culture medium with time tends to increase and decrease and tend to be stable, and the maximum biomass of the viable count after 48h of culture is 3.84 × 10 10 cfu/mL, the subsequent reduction in biomass may be due to the rapid availability of brown sugar as a rapid carbon source by microorganisms but the increasing availability of carbon and nitrogen with growthThe ratio change affects microbial growth resulting in a decrease in microbial biomass.
Therefore, compared with the single strain for fermenting the pig raising wastewater, the growth speed of the LZP-LZP-WB-SC composite strain in the pig raising wastewater is higher, the maximum biomass can be achieved within 48 hours, the fermentation period is greatly shortened, and the yield is improved.
Example 2 application of Single microbial inoculum in treatment of pig wastewater
1 materials and methods
1.1 sources of test strains
The same as in example 1.
1.2 physicochemical Properties of wastewater from pig raising
The swine wastewater used in the test was obtained from a certain pig farm with a flush Ha Ershi fural base region, and the basic characteristics of the collected swine wastewater are shown in table 2.1.3 culture Medium
Beef extract peptone medium: 10.00g of peptone, 3.00g of beef extract, 5.00g of NaCl, 20.00g of agar, and adding no agar into a liquid culture medium, wherein the volume of distilled water is up to 1000.00mL.
The pig raising wastewater culture medium after the carbon-nitrogen ratio is optimized: 10g of brown sugar and the volume of the pig raising wastewater is up to 1000.00mL.
1.4 methods of investigation
Beneficial microorganism strains LZP, LZP, WB and SC preserved at-80 ℃ are respectively inoculated into beef extract peptone solid culture media and cultured for 24h at 30 ℃ for activation. The strains after 2-3 rings of activation are inoculated in a 250mL triangular flask filled with 100mL beef extract peptone liquid culture medium by using an inoculating ring under the aseptic operation condition, shaking culture is carried out at 30 ℃ and 120r/min, then each strain cultured to the same concentration (OD 600= 1.0) is used as seed liquid, then activated bacterial liquids of LZP03, LZP, WB and SC are respectively inoculated in 300mL pig-raising wastewater culture medium according to the inoculation amount of 1vol%, then shaking culture is carried out at 30 ℃ and 120r/min, samples are sampled every 24h, centrifugation is carried out for 5min at 12000rpm, and then index determination is carried out.
1.5 test measurement index
1.5.1 Water quality index
And (4) measuring the contents of pH, total nitrogen, total carbon, total organic carbon, ammoniacal nitrogen and phosphorus of the sample.
1.5.2 microbiological indicators
Placing 10mL of fermentation liquor into 90mL of sterile water, shaking for 30min in a shaking table at 200r/min, and then performing gradient dilution to obtain 10 -7 、10 -8 、10 -9 And (3) taking 0.1mL of the diluent, coating the diluent on a beef extract peptone medium plate, culturing at 30 ℃ for 24h, calculating the number of viable bacteria (cfu/mL), and setting three groups in parallel.
2. Results and discussion
2.1 Change of Each index of fermentation waste liquid after removing thallus from pig-raising waste water treated by single bacterium agent
2.1.1 Change of Total Nitrogen, total carbon and Total organic carbon contents of Single microbial inoculum in optimized pig-raising wastewater
And (3) inoculating LZP into the optimized pig raising wastewater culture medium, and measuring total nitrogen, total carbon and total organic carbon of the centrifuged supernatant every 24 hours. As can be seen from fig. 2-4, the total nitrogen content in the centrifuged supernatant of LZP02 fermentation broth was reduced by 93.40%, the total carbon content was reduced by 84.72%, and the total organic carbon content was reduced by 89.86% compared to the control group.
And (3) inoculating LZP into the optimized pig raising wastewater culture medium, and measuring total nitrogen, total carbon and total organic carbon of the centrifuged supernatant every 24 h. As can be seen from fig. 2-4, the total nitrogen content in the supernatant of the LZP03 fermentation broth after centrifugation was decreased by 94.58%, the total carbon content was decreased by 85.96%, and the total organic carbon content was decreased by 89.62% compared to the control group.
And inoculating the SC strain into a pig raising wastewater culture medium with an optimized carbon-nitrogen ratio, and measuring total nitrogen, total carbon and total organic carbon of the centrifuged supernatant every 24 hours. As can be seen from fig. 2-4, the total nitrogen content in the supernatant of the SC strain fermentation broth after centrifugation is reduced by 94.23%, the total carbon content is reduced by 78.64%, and the total organic carbon content is reduced by 77.84% compared to the control group.
And (4) inoculating WB into the optimized pig raising wastewater culture medium, and measuring total nitrogen, total carbon and total organic carbon of the centrifuged supernatant every 24 hours. As can be seen from fig. 2-4, the total nitrogen content in the WB fermentation broth supernatant after centrifugation was reduced by 93.87%, the total carbon content was reduced by 75.53%, and the total organic carbon content was reduced by 75.06% compared to the control group.
The results show that the LZP, LZP, SC and WB strains can better utilize the nutrient components in the swine wastewater to perform self growth, reproduction and metabolic activities, and prevent the filtered fermentation wastewater from causing environmental pollution due to rich nutrition.
2.1.2 changes in pH and COD of the optimized wastewater culture Medium from pig farming
The pH measurement is carried out on the LZP02 fermentation liquor every 24h, and the pH of the pig raising wastewater is reduced from an alkaline solution with the original pH =8.85 to a neutral fermentation liquor with the pH =7.37 after the pig raising wastewater is fermented for 96h by the LZP strain, as can be seen from the graph in FIG. 5. In order to determine the effect of the screened strain on removing COD from the pig-raising wastewater, COD detection is carried out on the LZP02 fermentation liquid supernatant, and as can be seen from FIG. 6, with the continuous increase of the strain fermentation time, the COD content is continuously and rapidly reduced, the maximum COD removal rate reaches 92.25%, which indicates that LZP has a better removal rate on COD.
The pH measurement of the LZP03 fermentation liquor is carried out every 24h, and it can be seen from FIG. 5 that the pH of the pig-raising wastewater is reduced from the original alkaline solution with pH =8.85 to the neutral fermentation liquor with pH =7.34 after the pig-raising wastewater is fermented for 96h by the LZP strain. In order to determine the effect of LZP on removing COD from pig wastewater, COD detection is carried out on LZP03 fermentation liquid supernatant, and as can be seen from FIG. 6, with the continuous increase of the fermentation time of the strain, the COD content is continuously and rapidly reduced to 1280mg/L at most, and the COD removal rate reaches 92%, which indicates that LZP has a better removal rate for COD.
The SC strain fermentation liquid was subjected to pH measurement every 24h, and it can be seen from fig. 5 that after the pig wastewater was fermented by the SC strain for 96h, the pH of the pig wastewater was reduced from the original alkaline solution with pH =8.85 to the acidic fermentation liquid with pH = 6.31. In order to determine the effect of the screened strains on removing COD from the pig wastewater, COD detection is carried out on the supernatant of the SC strain fermentation liquor, and as can be seen from figure 6, with the continuous increase of the fermentation time of the strains, the COD content is continuously and rapidly reduced, and the maximum removal rate of COD reaches 81.25%, which indicates that the SC strain has better removal rate of COD.
The WB fermentation broth was subjected to pH measurement every 24h, and it can be seen from fig. 5 that the pH of the swine wastewater was reduced from the original alkaline solution with pH =8.85 to the acidic fermentation broth with pH =5.86 after 96h fermentation by the WB strain. In order to determine the effect of the screened strains on removing COD from the pig wastewater, COD detection is carried out on the supernatant of WB fermentation liquor, and as can be seen from FIG. 6, with the continuous increase of the fermentation time of the strains, the COD content is continuously and rapidly reduced, and the maximum removal rate of COD reaches 82%, which indicates that WB has a better removal rate of COD.
2.1.3 removal rate of ammoniacal nitrogen and phosphorus by single strain in optimized pig-raising wastewater culture medium
The determination of the ammonia nitrogen content in the supernatant of LZP in different fermentation times shows that LZP strain can greatly reduce the ammonia nitrogen content in the fermentation waste liquid, the removal rate of the ammonia nitrogen reaches 91.11%, and the removal effect of the ammonia nitrogen is good, as shown in fig. 7 and 8. The phosphorus content of the supernatant of the fermentation liquor is also measured, and compared with a control group LZP02, the phosphorus removal rate can reach 25.63%, and the phosphorus removal effect is general.
The determination of the ammonia nitrogen content in the supernatant of LZP fermented liquid at different fermentation times shows that LZP strain can greatly reduce the ammonia nitrogen content in the fermented waste liquid, the removal rate of the ammonia nitrogen reaches 90.35%, and the removal effect of the ammonia nitrogen is good, as shown in fig. 7 and 8. And the phosphorus content of the fermentation liquid supernatant is measured after the bacterial strain is fermented, so that whether the bacterial strain can well reduce the phosphorus content of the fermentation waste liquid can be seen. The graph shows that the phosphorus removal rate of LZP03 compared with a control group can reach 69.10%, which indicates that the phosphorus removal rate has certain phosphorus removal capacity.
The ammonia nitrogen content of the fermentation liquor supernatant of the SC strain in different fermentation times is measured, and as can be seen from figures 7 and 8, the SC strain can obviously reduce the ammonia nitrogen content in the fermentation waste liquor, the removal rate of the ammonia nitrogen reaches 89.98%, and the removal effect of the ammonia nitrogen is good. Phosphorus content determination of the fermentation liquor supernatant is carried out, and the phosphorus removal rate of the SC strain can reach 17.39% compared with that of a control group, so that the SC strain has a certain phosphorus removal effect.
The ammonia nitrogen content of the fermentation liquor supernatant of WB at different fermentation times is measured, and it can be known from figures 7 and 8 that the WB strain can significantly reduce the ammonia nitrogen content of the fermentation waste liquor, the removal rate of the ammonia nitrogen reaches 76.33%, and the removal effect of the ammonia nitrogen is good. Phosphorus content determination of the supernatant of the fermentation liquor is carried out, and the phosphorus removal rate can reach 35.88% compared with that of WB of a control group, so that the phosphorus removal effect is achieved.
Example 3 application of Complex microbial Agents in treatment of wastewater from pig farming
1 materials and methods
1.1 sources of test strains
The same as in example 1.
1.2 physicochemical Properties of wastewater from pig raising
The swine wastewater used in the experiment is taken from a certain pig farm in the homogeneous Ha Ershi Fulal base region, 1wt% of brown sugar is added into the collected swine wastewater to optimize the carbon-nitrogen ratio to be used as a basic fermentation culture medium, and the basic characteristics of the optimized swine wastewater are shown in Table 3.
TABLE 3 optimization of post-pig-raising wastewater physicochemical properties
1.3 Medium
Beef extract peptone medium: 10.00g of peptone, 3.00g of beef extract, 5.00g of NaCl, 20.00g of agar, and adding no agar into a liquid culture medium, wherein the volume of distilled water is up to 1000.00mL.
1.4 methods of investigation
Beneficial microorganism strains LZP, LZP, WB and SC which are preserved at-80 ℃ are respectively inoculated into a beef extract peptone solid medium and cultured for 24h at 30 ℃ for activation. The strains after 2-3 rings activation were inoculated in a 250mL triangular flask containing 100mL beef extract peptone liquid medium under aseptic conditions using an inoculating loop, and subjected to shaking culture at 30 ℃ and 120r/min, then each strain cultured to the same concentration (OD 600= 1.0) was used as a seed solution, and then activated bacteria solutions of LZP, LZP, WB, SC were mixed at a volume ratio of 1.
1.5 test measurement index
1.5.1 Water quality index
And (4) measuring the contents of pH, total nitrogen, total carbon, total organic carbon, ammoniacal nitrogen and phosphorus of the sample.
1.5.2 microbiological indicators
Placing 10mL of fermentation liquor into 90mL of sterile water, shaking for 30min in a shaking table at 200r/min, and then performing gradient dilution to obtain 10 -7 、10 -8 、10 -9 And (3) taking 0.1mL of the diluent, coating the diluent on a beef extract peptone medium plate, culturing at 30 ℃ for 24h, calculating the number of viable bacteria (cfu/mL), and setting three groups in parallel.
2. Results and discussion
2.1 Change of Each index of fermentation waste liquid after removing thallus from pig-raising waste water treated by composite microbial agent
2.1.1 changes of total nitrogen, total carbon and total organic carbon contents of the compound microbial inoculum in the optimized pig-raising wastewater
And inoculating the compound microbial agent to an optimized culture medium, and measuring the total nitrogen of the fermentation liquor every 24 hours. As can be seen from FIG. 9, the total nitrogen content of the fermentation broth inoculated with the complex microbial inoculant in the optimized culture medium shows a general decreasing trend along with the prolonging of the fermentation time, and rapidly decreases in the fermentation interval of 0-24h, then gradually decreases, and reaches a minimum value of 102.2mg/L when the fermentation broth is cultured for 96h, which is 73.55% less than that of the control group.
The change of the total carbon of the complex microbial inoculant is also obviously reduced as shown in FIG. 9. The total carbon content in the fermentation liquor shows a slow descending trend along with the increase of the culture time, the minimum value of the total carbon content in the fermentation liquor of the compound microbial agent is 3010mg/L, compared with a control group, the total carbon reduction rate of the fermentation liquor of the compound microbial agent after 96 hours of culture is 48.55%, and the reduction of the total carbon content is probably caused by the fact that the total carbon content is reduced due to the fact that carbon dioxide is discharged by continuous aerobic respiration of microorganisms in the growth process, which indicates that the microorganisms can actively grow and reproduce in the pig wastewater. In the aspect of total organic carbon, the compound microbial inoculum is inoculated in the optimized pig-raising wastewater culture medium, the total organic carbon content of the fermentation liquor continuously decreases and finally tends to be stable along with the prolonging of the culture time, the total organic carbon content of the centrifugal fermentation liquor rapidly decreases to reach the minimum value in 48 hours and then tends to be stable after slightly increasing in 72 hours in the fermentation period of 0-24 hours, and the minimum decrease rate of the total organic carbon of the compound microbial inoculum compared with a control group reaches 54.10%.
2.1.2 pH Change of Complex microbial Agents in optimized pig wastewater
The pH value of the fermentation liquor of the compound microbial agent is measured every 24 hours, and as can be seen from fig. 10, after the pig-raising wastewater is fermented for 96 hours by the compound microbial agent, the pH value of the pig-raising wastewater is reduced from an original weak alkaline solution with pH =7.82 to a weak acidic fermentation liquor with pH =6.51, which is probably due to the fact that the thalli secrete organic acids, the pH value is reduced, and the method has important significance for threat prevention of the pig-raising wastewater to soil salinization.
2.1.3 change of COD content and removal rate of compound microbial inoculum in optimized pig-raising wastewater
In order to determine the effect of the composite microbial inoculum on removing COD from the swine wastewater, COD detection is carried out on supernatant of fermentation liquid, as can be seen from figure 11, with the continuous increase of the fermentation time of the bacterial strain, the COD content is continuously and rapidly reduced from original COD =27400mg/L to COD =9360mg/L at the maximum, the removal rate of the COD reaches 65.38%, which indicates that the composite microbial inoculum has a certain removal rate on the COD from the swine wastewater, and the threat of filtered waste liquid to the environment is reduced.
2.1.4 removal rate of ammonia nitrogen and phosphorus in optimized pig raising wastewater by using compound microbial inoculant
The ammonia nitrogen content of the fermentation liquid supernatant of the compound microbial inoculum in different fermentation time is measured, and as can be seen from fig. 12, the compound microbial inoculum can greatly reduce the ammonia nitrogen content of the fermentation waste liquid, the removal rate of the ammonia nitrogen reaches 58.17%, and the removal effect of the ammonia nitrogen is good. After the fermentation, the phosphorus content of the supernatant of the fermentation liquor is measured, so that whether the strain can well reduce the phosphorus content removal rate in the fermentation waste liquor can reach 44.77 percent or not can be seen, and the strain has good phosphorus removal capability.
Example 4 production of beneficial monobacteria Strain LZP (Bacillus megaterium) powder Using pig wastewater
The method comprises the following specific steps:
step one, strain activation: the method comprises the steps of inoculating LZP preserved at minus 80 ℃ into a liquid culture medium, inoculating 2-3 rings of the preserved strain in a centrifuge tube into a 250mL triangular flask filled with 100mL of the liquid culture medium under the aseptic operation condition by using an inoculating loop, and performing shake culture at 30 ℃ for 24 hours at 180r/min for activation to obtain an activated bacterial liquid for later use.
The liquid culture medium comprises the following components: according to the mass percentage, the liquid culture medium contains 0.3 percent of beef extract, 1 percent of peptone, 0.5 percent of sodium chloride and the balance of water;
step two, seed liquid culture: inoculating the LZP03 liquid activated bacterium solution prepared in the first step into a 2L triangular flask filled with 500mL of seed culture medium according to the inoculation amount of 1vol%, and performing shake culture at 34 ℃ for 24h at 180r/min for seed liquid culture to obtain seed liquid for later use.
The seed culture medium comprises the following components: according to the mass percentage, the seed culture medium contains 1 percent of brown sugar, and the rest is pig-raising wastewater.
Step three, first-stage seed fermentation tank culture: inoculating the seed solution cultured in the second step into a 50L fermentation tank containing 50L fermentation medium, wherein the inoculation amount is 1% of the total volume of the fermentation medium, culturing at 35 deg.C for 24h at 150r/min, and introducing sterile air continuously until the viable count reaches 3 × 10 10 And obtaining seed fermentation liquor for later use after cfu/mL.
The fermentation medium comprises the following components: according to the mass percentage, the fermentation medium contains 1 percent of brown sugar, and the rest is the pig-raising wastewater.
Step four, secondary fermentation tank amplification culture: inoculating the seed fermentation liquid cultured in the third step into 500L fermentation tank containing 500L fermentation medium, wherein the inoculation amount is 10% of the volume of the fermentation tank, culturing at 38 deg.C for 24 hr at 100r/min, and introducing sterile air continuously until viable count reaches 3 × 10 10 And obtaining seed fermentation liquor for later use after cfu/mL.
And the components of the fermentation medium are consistent with those of the fermentation medium in the third step.
Step five, third-stage fermentation tank expanded culture: inoculating the fermentation broth cultured in the fourth step into a third-stage fermentation tank filled with 3000L of fermentation medium, wherein the inoculation amount is 5% of the volume of the fermentation tank, culturing at 39 deg.C and 80r/min for 48h, continuously introducing sterile air, and measuring the viable count to obtain 4.64 × 10 10 And (5) cfu/mL, stopping culturing, and discharging to obtain fermentation liquor.
And the components of the fermentation medium are consistent with those of the fermentation medium in the third step.
Step six, bacterial liquid flocculation: adding a biological flocculant into the obtained fermentation liquor, quickly stirring at 200r/min for 10min, slowly stirring at 30r/min for 30min, and standing for 60min to obtain a settled thallus flocculation liquid.
Step seven, plate and frame filter pressing: and (4) uniformly mixing the settled thallus flocculation liquid obtained in the sixth step at 150r/min, and filtering by using a plate-and-frame filter press to obtain flocculated wet bacteria.
Step eight, drying the raw powder: and (3) putting the wet thalli obtained after the filter pressing in the step seven into a vertical boiling dryer for drying, wherein the drying temperature is 40 ℃, and the drying time is 6 hours.
Ninth, beneficial microorganism powder preparation: mixing the beneficial microorganism raw powder obtained in the step eight with anhydrous calcium carbonate, wherein the mixing ratio is 1:15, stirring in a stirrer for 0.5-2h, crushing, and sieving with a 100-mesh sieve to obtain a viable count of 8 × 10 10 -1×10 11 cfu/g of beneficial single strain LZP bacterial powder.
Example 5 production of beneficial monobacterial Strain LZP (Bacillus pumilus) bacterial powder Using pig wastewater
The specific procedure was as in example 4, except that beneficial monobacterium strain LZP (Bacillus megaterium) was replaced with beneficial monobacterium strain LZP (Bacillus pumilus). The final viable count is 8 × 10 10 -1×10 11 cfu/g of beneficial single strain LZP bacterial powder.
Example 6 production of beneficial Single-Strain WB (Bacillus velezensis) powder Using pig-raising wastewater
The specific procedure is as in example 4, except that a beneficial single strain L is introducedZP03 (Bacillus megaterium) is replaced by a beneficial single-bacterium strain WB (Bacillus velezensis). The final viable count is 7 × 10 10 -9×10 10 cfu/g of beneficial single-strain WB strain powder.
Example 7 production of beneficial Single Strain SC (Bacillus subtilis) powder Using pig wastewater
The specific procedure is as in example 4, except that beneficial monobacterium strain LZP (Bacillus megaterium) is replaced with beneficial monobacterium strain SC (Bacillus subtilis). The final viable count is 7 × 10 10 -9×10 10 cfu/g of beneficial single strain SC bacterial powder.
Example 8 production of Complex microbial Agents Using wastewater from pig Breeding
The process flow chart is shown in fig. 13, and the specific steps are as follows:
step one, strain activation: beneficial microorganism strains LZP (Bacillus megaterium), LZP (Bacillus pumilus), WB (Bacillus velezensis) and SC (Bacillus subtilis) preserved at-80 ℃ are respectively inoculated into a liquid culture medium, the preserved strains in a centrifugal pipe are inoculated into 2-3 rings of 250mL triangular bottles filled with 100mL of liquid culture medium by using inoculating rings under the aseptic operation condition, and the flask is subjected to shake culture for 24 hours at 180r/min under the 30 ℃ condition to obtain activated bacteria liquid for later use.
The liquid culture medium comprises the following components: according to the mass percentage, the liquid culture medium contains 0.3 percent of beef extract, 1 percent of peptone, 0.5 percent of sodium chloride and the balance of water;
step two, seed liquid culture: taking LZP, LZP, WB and SC liquid activated bacteria liquid prepared in the first step, and mixing the liquid activated bacteria liquid with the liquid activated bacteria liquid according to the ratio of 1:1, mixing, inoculating the mixture into a 2L triangular flask filled with 500mL of seed culture medium according to the inoculation amount of 1vol%, and then performing shake culture at 34 ℃ for 24h at 180r/min for seed liquid culture to obtain seed liquid for later use.
The seed culture medium comprises the following components: according to the mass percentage, the seed culture medium contains 1 percent of brown sugar, and the rest is pig-raising wastewater.
Step three, first-stage seed fermentation tank culture: inoculating the seed solution cultured in the second step into 50L of fermentation mediumInoculating in a fermenter with an inoculum size of 1% of the total volume of the fermentation medium, culturing at 35 deg.C for 24 hr at 100r/min, and introducing sterile air until viable count reaches 3 × 10 10 And obtaining seed fermentation liquor for later use after cfu/mL.
The fermentation medium comprises the following components: according to the mass percentage, the fermentation medium contains 1 percent of brown sugar, and the balance is the pig-raising wastewater.
Step four, secondary fermentation tank amplification culture: inoculating the seed fermentation liquid cultured in the third step into 500L fermentation tank containing 500L fermentation medium, wherein the inoculation amount is 10% of the volume of the fermentation tank, culturing at 38 deg.C for 24 hr at 100-150r/min, and introducing sterile air continuously until viable count reaches 3 × 10 10 And obtaining seed fermentation liquor for later use after cfu/mL.
And the components of the fermentation medium are consistent with those of the fermentation medium obtained in the third step.
Step five, third-stage fermentation tank expanded culture: inoculating the fermentation broth cultured in the fourth step into a three-stage fermentation tank filled with 3000L of fermentation medium, wherein the inoculation amount is 5% of the volume of the fermentation tank, culturing at 39 deg.C and 80r/min for 48h, and introducing sterile air continuously during the period until the final viable count reaches 4.86 × 10 10 Stopping culturing after cfu/mL or more, and discharging to obtain fermentation liquor.
And the components of the fermentation medium are consistent with those of the fermentation medium obtained in the third step.
Step six, bacterial liquid flocculation: adding bioflocculant into the obtained fermentation liquor, quickly stirring at 200r/min for 10min, slowly stirring at 30r/min for 30min, and standing for 60min to obtain settled thallus flocculation liquid.
Step seven, plate and frame filter pressing: and (4) uniformly mixing the settled thallus flocculation liquid obtained in the sixth step at 150r/min, and filtering by using a plate-and-frame filter press to obtain flocculated wet bacteria.
Step eight, drying the raw powder: and (3) putting the wet thalli obtained after the filter pressing in the step seven into a vertical boiling dryer for drying, wherein the drying temperature is 40 ℃, and the drying time is 6 hours.
Ninth, beneficial microorganism powder preparation: will step withMixing the beneficial microorganism raw powder obtained in the eighth step with anhydrous calcium carbonate, wherein the mixing ratio is 1:15-20, stirring in a stirrer for 0.5-2h, pulverizing, sieving with 100 mesh sieve to obtain viable bacteria with viable count of 8 × 10 10 -1×10 11 cfu/g of compound microbial powder.
Example 9 application of Complex microbial preparation to promotion of plant growth
Diluting the compound microbial powder prepared in the example 8 by using sterile water, wherein the dilution multiple is 200 times, fertilizing the Chinese cabbage by using the diluted bacterial liquid in a spraying manner, wherein the spraying amount is 1-2 mu of land, one barrel (25 kg) of the diluted bacterial agent is applied, the application is performed once every ten days, the watering is performed normally at other times, leaf spraying is performed on the Chinese cabbage in the seedling stage and the rosette stage respectively, and the dilution multiple of the bacterial agent is 300 times during the leaf spraying. The test time is 30d, and the control group is watered only and does not apply the microbial inoculum.
The results are shown in fig. 14, and show that the Chinese cabbage using the microbial inoculum has developed root system, good growth vigor and obvious yield increase effect compared with the control group.
Claims (6)
1. A method for producing a beneficial microbial agent by using pig raising wastewater while removing environmental pollutants in the pig raising wastewater is characterized by comprising the following steps:
step one, strain activation: inoculating beneficial microorganisms preserved at-80 ℃ into a liquid culture medium, inoculating the preserved strains in a centrifuge tube into a 250mL triangular flask filled with 100mL of liquid culture medium by using an inoculating loop under the aseptic operation condition, and performing shake culture at 30 ℃ for 24 hours at 180r/min for activation to obtain activated bacterial liquid for later use;
wherein the liquid culture medium comprises the following components: according to the mass percentage, the liquid culture medium contains 0.3 to 0.4 percent of beef extract, 1 to 1.2 percent of peptone, 0.5 percent of sodium chloride and the balance of water;
step two, seed liquid culture: inoculating the liquid activated bacterium liquid prepared in the first step into a 2L triangular flask filled with 500mL of seed culture medium according to the inoculation amount of 1-3 vol%, and performing shake culture at 34 ℃ at 180r/min for 24h to obtain a seed solution for later use;
the seed culture medium comprises the following components: according to the mass percentage, the seed culture medium contains 1 to 1.5 percent of brown sugar, and the rest is pig raising wastewater;
step three, first-stage seed fermentation tank culture: inoculating the seed solution cultured in the second step into a 50L fermentation tank containing 50L fermentation medium, wherein the inoculation amount is 1% of the total volume of the fermentation medium, culturing at 35-38 deg.C for 24-36h at 100-150r/min, and introducing sterile air continuously until the viable count reaches 2 × 10 10 -4×10 10 Obtaining seed fermentation liquor for later use after cfu/mL;
the fermentation medium comprises the following components: according to the mass percentage, the fermentation medium contains 1 to 1.5 percent of brown sugar, and the rest is pig-raising wastewater;
step four, secondary fermentation tank amplification culture: inoculating the seed fermentation liquid cultured in the third step into a 500L fermentation tank containing 500L fermentation medium, wherein the inoculation amount is 5% -10% of the volume of the fermentation tank, culturing at 35-38 ℃ at 100-150r/min for 24h, continuously introducing sterile air during the period, and when the viable count reaches 2 x 10 10 -4×10 10 Obtaining seed fermentation liquor after cfu/mL for later use;
the components of the fermentation medium are consistent with those of the fermentation medium in the third step;
step five, third-stage fermentation tank expanded culture: inoculating the fermentation liquid cultured in the fourth step into a three-stage fermentation tank filled with 3000L of fermentation medium, wherein the inoculation amount is 5% -10% of the volume of the fermentation tank, culturing at 37-39 deg.C and 60-80r/min for 24-48 h, continuously introducing sterile air during the period, and when the viable count reaches 4 × 10 10 Stopping culturing after cfu/mL is above, and discharging to obtain fermentation liquor;
the components of the fermentation medium are consistent with those of the fermentation medium in the third step;
step six, bacterial liquid flocculation: adding a bioflocculant into the obtained fermentation liquor, stirring and standing to obtain a settled thallus flocculation liquid;
step seven, plate and frame filter pressing: uniformly mixing the settled thallus flocculation liquid obtained in the sixth step at 150r/min, and filtering by using a plate-and-frame filter press to obtain flocculated wet bacteria and fermentation waste liquid;
step eight, drying the raw powder: putting the wet thalli obtained after filter pressing in the seventh step into a vertical boiling dryer for drying, wherein the drying temperature is 40 ℃, and the drying time is 6 hours;
ninth, beneficial microorganism powder preparation: mixing the beneficial microorganism raw powder obtained in the step eight with anhydrous calcium carbonate, wherein the mixing mass ratio is 1:15 to 20, stirring for 0.5 to 2 hours in a stirrer, and sieving with a 100-mesh sieve after crushing to obtain the beneficial microbial powder.
2. The method of claim 1, wherein the beneficial microorganisms comprise: LZP03 (Bacillus megaterium), LZP (Bacillus pumilus), WB (Bacillus velezensis) and SC (Bacillus subtilis), wherein the beneficial microorganisms are all preserved in China center for type culture Collection with the preservation numbers of CCTCC NO: m2018599, CCTCC NO: m2018598, CCTCC NO: m20211547, CCTCC NO: m20211546.
3. The method according to claim 1, wherein the beneficial microorganism consists of LZP (Bacillus megaterium), LZP (Bacillus pumilus), WB (Bacillus velezensis), SC (Bacillus subtilis) in a volume ratio of 1.
4. The method as claimed in claim 1, wherein the bacterial liquid in the sixth step is flocculated by adding a bioflocculant into the obtained fermentation liquid, stirring at 200r/min for 10min, stirring at 30r/min for 30min, and standing for 60min to obtain a settled thallus flocculation liquid.
5. A beneficial microbial powder prepared according to the method of any one of claims 1-4.
6. Use of the beneficial microbial inoculant powder of claim 5 for crop growth promotion and crop biocontrol.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210108012.0A CN115287208A (en) | 2022-01-28 | 2022-01-28 | Method for producing beneficial microbial agent by using pig raising wastewater while removing environmental pollutants in pig raising wastewater |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210108012.0A CN115287208A (en) | 2022-01-28 | 2022-01-28 | Method for producing beneficial microbial agent by using pig raising wastewater while removing environmental pollutants in pig raising wastewater |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115287208A true CN115287208A (en) | 2022-11-04 |
Family
ID=83820394
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210108012.0A Withdrawn CN115287208A (en) | 2022-01-28 | 2022-01-28 | Method for producing beneficial microbial agent by using pig raising wastewater while removing environmental pollutants in pig raising wastewater |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115287208A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115558657A (en) * | 2022-11-23 | 2023-01-03 | 山东义才和锐生物技术有限公司 | Process for preparing cosmetic auxiliary material by fermenting marine microorganisms |
-
2022
- 2022-01-28 CN CN202210108012.0A patent/CN115287208A/en not_active Withdrawn
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115558657A (en) * | 2022-11-23 | 2023-01-03 | 山东义才和锐生物技术有限公司 | Process for preparing cosmetic auxiliary material by fermenting marine microorganisms |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102199562B (en) | Composite bacterial preparation as well as preparation method and application thereof | |
| CN107586745B (en) | Livestock manure compost deodorization nitrogen-retention strain, microbial inoculum and preparation method and application thereof | |
| CN113337418B (en) | Functional strain W25 capable of reducing heavy metal content of vegetables and improving quality of vegetables and application | |
| CN109022327B (en) | Preparation method of microbial mixed inoculant and application of microbial mixed inoculant in high-temperature composting | |
| CN115353986B (en) | Bacillus bailii strain WB strain for treating pig raising wastewater and application thereof | |
| CN109110912A (en) | A kind of dirt collecting pit pool black and odorous water administering method | |
| CN107177533A (en) | A kind of Thermophilic Bacteria compounding microbial inoculum and preparation method and application | |
| CN110002611A (en) | A kind of breeding water body regulator and preparation method thereof | |
| Cai et al. | Compost-derived indole-3-acetic-acid-producing bacteria and their effects on enhancing the secondary fermentation of a swine manure-corn stalk composting | |
| CN104789494B (en) | The method for improving turf salt-resistance using garbage compost microbial bacterial agent is strengthened | |
| CN115287208A (en) | Method for producing beneficial microbial agent by using pig raising wastewater while removing environmental pollutants in pig raising wastewater | |
| CN115353210B (en) | Application of bacillus pumilus LZP02 in treatment of pig raising wastewater | |
| CN115353987B (en) | Bacillus subtilis strain SC strain for treating pig raising wastewater and application thereof | |
| CN115353211B (en) | Application of bacillus megatherium LZP03 in treatment of pig raising wastewater | |
| CN114752538B (en) | Oil tea endophyte with soil improvement function and application thereof | |
| CN115959931A (en) | Method for preparing organic fertilizer by deep fermentation of livestock and poultry manure and straws | |
| CN115287209B (en) | Composite microbial agent and application thereof in treating swine waste water | |
| CN112919982A (en) | Compound microbial agent and fertilizer special for eucalyptus | |
| CN110358701B (en) | Paenibacillus polymyxa microbial agent and production method thereof | |
| CN117603888B (en) | Bacillus cereus and application thereof in cultivation tail water treatment | |
| Xu et al. | Characterization of microorganisms in the soils with sewage irrigations | |
| CN117431166B (en) | Novel trichoderma brevicompactum strain TB2 and application thereof | |
| CN117229983B (en) | Nitrosogenic paraburkholderia PX-418, microbial inoculum and application thereof | |
| CN112342164B (en) | Rhodoblastus sphagnicola coupled microbial fertilizer and preparation and application thereof | |
| CN111995455A (en) | Production method of blue algae, bacterial slag and rice hull selenium-rich organic fertilizer |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20221104 |
|
| WW01 | Invention patent application withdrawn after publication |