CN115353211A - Application of bacillus megaterium LZP03 in treatment of pig raising wastewater - Google Patents
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Images
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- 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
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/105—Phosphorus compounds
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/22—Nature of the water, waste water, sewage or sludge to be treated from the processing of animals, e.g. poultry, fish, or parts thereof
Abstract
The invention discloses application of bacillus megaterium LZP03 in treatment of pig raising wastewater. The bacillus megaterium LZP03 is preserved in China center for type culture collection, and the strain preservation number is CCTCC NO. M2018599. According to the invention, different kinds of beneficial microorganisms are inoculated to the pig raising wastewater to screen and obtain the bacillus megaterium LZP03 which grows in the pig raising wastewater and has a higher propagation speed, and the utilization condition of the bacillus megaterium LZP03 on nutrient components in the pig raising wastewater and the degradation condition of pollutants in the pig raising wastewater are detected, so that the result shows that the LZP03 can grow in an original pig raising wastewater culture medium and has higher biomass, and the biomass is obviously improved after the pig raising wastewater culture medium with optimized carbon-nitrogen ratio is used for inoculating the LZP 03. The LZP03 strain is fermented in the pig raising wastewater culture medium with the optimized carbon-nitrogen ratio to remove COD, ammonia nitrogen and phosphorus well.
Description
Technical Field
The invention relates to a microbial strain capable of being used for treating pig raising wastewater, in particular to application of bacillus megatherium LZP03 in treating pig raising wastewater, and also relates to a method for treating pig raising wastewater. The invention belongs to the technical field of agricultural production.
Background
China is one of the most huge countries of the large-scale breeding and consumption industries of live pigs in the world. With the continuous development of economy, the scale of the live pig breeding industry chain is continuously enlarged, according to the annual book of statistics in China in 2016, the annual output of pork in 2015 is 5486.5 ten thousand tons, the live pig breeding scale is very large (the national statistics bureau, 2016), and the produced livestock excrement also becomes a main pollution source, so that the problem of environmental pollution caused by the pollution is more and more prominent. The total production of livestock and poultry manure in China now reaches 2.43x10 8 Ton, urine is 1.63x10 8 Ton, total nitrogen 1.02x10 6 Ton, total phosphorus 1.60x10 5 The discharge amount of COD of the pig farm is 7118 million tons (Panqing, 2002), which is far more than the sum of the discharge amounts of industrial wastewater and domestic wastewater COD, and as most of pig farms adopt flushing water to clean pig houses, the pig urine, excrement and the like in the pig farm wastewater are mixed with each other, the ammonia nitrogen content, phosphorus content, suspended matters and organic matters in the wastewater are high, and the carbon-nitrogen ratio is seriously unbalanced, and if a large amount of untreated discharge is carried out in lakes, rivers, farmlands and other environments, the problems of water eutrophication, soil property change and other pollution are caused.
At present, the pig-raising wastewater treatment technology at home and abroad mainly focuses on three main modes, namely a physicochemical treatment technology, a natural treatment technology and a biological treatment technology. The physical and chemical treatment methods mainly used include a medium adsorption method, a flocculation precipitation method and the like, for example, qianfeng and the like utilize zeolite-straw combination to filter swine urine wastewater, the removal rates of COD, ammoniacal nitrogen and phosphorus can respectively reach 47.9%, 72.9% and 50.1% (Qianfeng, 2008), and like Traine and the like, 61.02% (Traine, 2010) of swine wastewater with the COD concentration of 3232mg/L can be removed by a magnetic flocculation method. Although the physicochemical method has better pollutant removal rate on the pig urine wastewater, the physicochemical method has the disadvantages of lower broad spectrum for removing the pig urine wastewater pollution, higher pollutant removal cost, larger investment on treatment equipment, less engineering application and the like. The natural treatment method generally utilizes natural soil, water and organisms to carry out comprehensive utilization for removing pollutants, such as Zhu Xizhen and the like, and the removal rates of COD, BOD and phosphorus respectively reach 71-88%, 80-89% and 70-85% (Zhu Xizhen, 2003) by carrying out constructed wetland construction through substrates such as blast furnace slag and quartz sand, and Luxiu and the like utilize oxidation ponds to treat pig-raising wastewater COD less than or equal to 400mg/L and ammoniacal nitrogen less than or equal to 70mg/L (Luxiu, 2009). The natural treatment method has good decontamination capability for polluted wastewater, but has insufficient stability and long purification time. The biological treatment technology is a novel method for treating high-concentration organic wastewater by using the catalytic action of microorganisms, such as the Liangmeidong and the like, wherein the removal rate of COD (chemical oxygen demand) can be increased to more than 90% by using an SRB reactor for aeration treatment of pig raising wastewater (the Liangmeidong, 2009), and the Lifeng and the like can enable the removal rate of ammonia nitrogen and total nitrogen to reach 99.7% and 50.7% by using an aerobic-anaerobic combined treatment method (the Lifeng, 2011). However, in the process of treating wastewater by biological treatment, a large amount of activated sludge is generated and cannot be treated, so that secondary pollution is caused to the environment.
However, although the pig raising wastewater causes a series of pollution to the environment, the pig raising wastewater contains a large amount of organic matters, nitrogen, phosphorus, potassium and other nutrient elements which are also necessary for the growth of microorganisms, and has positive significance for improving the quantity and quality of the microorganisms. If the pig-raising wastewater is used as a basic culture medium for beneficial microbial fermentation production, the production cost of the microbial fertilizer can be greatly reduced, and the pollution problem can be reduced through biodegradation, so that the wastewater can be changed into valuable, the residual value of the wastewater can be 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.
Therefore, the method takes the pig wastewater as a natural screening culture medium, screens different beneficial microorganism strains to obtain strains which grow vigorously and propagate fast in the pig wastewater, optimizes nutrient components of the pig wastewater culture medium to improve strain biomass, detects the utilization condition of the strains on the nutrient components in the pig wastewater and the removal effect of pollutants after optimization, explores a fermentation system and a process of the pig wastewater, and provides technical support for harmless resource utilization of the pig wastewater.
Disclosure of Invention
One of the purposes of the invention is to provide a microbial strain which can be used for treating pig-raising wastewater;
the invention also aims to provide a method for treating the pig-raising wastewater.
In order to achieve the purpose, the invention adopts the following technical means:
according to the invention, different kinds of beneficial microorganisms are inoculated to the pig raising wastewater to screen and obtain a Bacillus megaterium LZP03 (CCTCC NO: M2018599) strain which grows in the pig raising wastewater and has a higher propagation speed, the utilization condition of the Bacillus megaterium LZP03 (CCTCC NO: M2018599) strain on nutrient components in the pig raising wastewater and the degradation condition of pollutants in the pig raising wastewater are detected, and the result shows that the LZP03 can grow in the original pig raising wastewater and has higher biomass, the biomass is obviously improved in the pig raising wastewater with an optimized carbon-nitrogen ratio, wherein the highest viable count of the LZP03 is 4.26 multiplied by 10 10 cfu/mL, and through comparing the LZP03 bacterial strain in the original and optimization pig raising waste water utilization carbon, nitrogen, organic carbon conditions prove in the optimization culture medium more vigorous, LZP03 bacterial strain in the optimization carbon nitrogen ratio of pig raising waste water culture medium fermentation to COD, ammonia nitrogen and phosphorus content all have better removal rate, COD removal rate reaches 92%, ammonia nitrogen removal rate is 90.35%, phosphorus removal rate is 69.10%, and can effectively adjust the original pig raising waste water pH value, reduced fermentation waste liquid to the environmental pollution possibility, better promotion pig raising waste water utilization carbon, nitrogen, organic carbon conditionsAnd (5) resource utilization of the wastewater.
On the basis of the research, the invention firstly provides the application of Bacillus megaterium LZP03 in treating the pig-raising wastewater, wherein the Bacillus megaterium LZP03 is named as Bacillus megaterium LZP03, is classified and named as Bacillus megaterium LZP03 respectively, is preserved in China center for type culture collection, has the strain preservation number of CCTCC NO. M2018599 in the university of Wuhan, china, and has the preservation time of 2018, 9 months and 6 days.
Preferably, the seed solution of the bacillus megaterium LZP03 is inoculated into the sterilized wastewater to be treated for pig raising, and the treated wastewater for pig raising can be obtained through fermentation, wherein the content of pollutants in the treated wastewater for pig raising is reduced compared with that in the prior treatment.
Preferably, the method also comprises the step of adding brown sugar into the sterilized pig raising wastewater to be treated, so that the carbon-nitrogen ratio of the pig raising wastewater is 16-20:1.
wherein, preferably, the fermentation refers to fermentation culture for 24-96h at 30 ℃ and 120 r/min.
Preferably, the pig raising wastewater after treatment has reduced contents of total nitrogen, total carbon, total organic carbon, COD, ammoniacal nitrogen and phosphorus compared with the pig raising wastewater before treatment.
Further, the invention also provides a method for treating the pig raising wastewater, which comprises the following steps:
(1) Inoculating Bacillus megaterium LZP03 into beef extract peptone culture medium for activation, inoculating activated strain into beef extract peptone culture medium, and regulating strain concentration to OD with sterile water 600 =1.0 as seed liquid; the bacillus megatherium LZP03 is preserved in China center for type culture collection, and the strain preservation number is CCTCC NO. M2018599;
(2) Inoculating the seed solution into sterilized wastewater to be treated for pig raising, and fermenting to obtain the treated wastewater for pig raising, wherein the content of pollutants in the treated wastewater for pig raising is reduced compared with that before treatment.
Preferably, the step (2) further comprises the step of adding brown sugar into the sterilized pig raising wastewater to be treated, so that the carbon-nitrogen ratio of the pig raising wastewater is 16-20:1.
wherein, in the step (2), the seed solution is preferably inoculated into the sterilized swine wastewater to be treated in an inoculation amount of 1-2 vol%.
Wherein, the fermentation in the step (2) is preferably carried out for 24-96h at 30 ℃ and 120 r/min.
Preferably, the treated swine wastewater has a reduced content of total nitrogen, total carbon, total organic carbon, COD, ammoniacal nitrogen and phosphorus compared with the swine wastewater before treatment.
Compared with the prior art, the invention has the beneficial effects that:
the invention realizes resource utilization of pig raising wastewater by using beneficial microorganisms, and the technology is also based on a biological treatment method, and has the core that the pollutant of the pig raising wastewater is removed by using a microbial aerobic-anaerobic combined treatment method, and the mass propagation of the beneficial microorganisms is realized by creatively using rich nutrition in the pig raising wastewater to obtain beneficial microorganism bacteria for preparing a microorganism bacteria agent, so that the pollutant is removed, the environment is protected, and the economic value is created. The invention takes the waste water of raising pigs as the natural culture medium to cultivate the microorganism, so the waste water of raising pigs is selected because the nutrient substances such as a large number of organic matters, carbon sources, nitrogen sources and the like contained in the waste water of raising pigs can provide sufficient nutrients for the growth of the microorganism, and the waste water of raising pigs can absorb, convert and fix the substances harmful to the environment in the waste water of raising pigs in the growth process of the microorganism, and can be used for the proliferation of the thallus and the secretion of metabolites, thus on one hand, reducing the sources of environmental safety threat pollution in the waste water of raising pigs such as ammoniacal nitrogen, organic matters, phosphorus and the like, on the other hand, a large number of microorganism thallus cultured by the waste water of raising pigs can collect the microbial preparations used in various aspects of plant promotion, crop biocontrol, pollutant degradation and the like, achieving two purposes at one stroke, and the higher cost of the culture medium of the microorganism in the culture process is one of the main reason of causing the price of the microbial inoculum to be higher, while the waste water of raising pigs is used to culture beneficial microorganism strains on the viable count close to the traditional culture medium, but the cost is greatly reduced, and has important significance for realizing green reduction of the use of the microbial fertilizer.
The invention carries out carbon nitrogen ratio optimization adjustment on the pig raising wastewater, inoculates beneficial microorganisms, finally obtains 1 beneficial microorganism LZP03 with higher viable count through screening, and has excellent effect on crop growth promotion, wherein the highest viable count of the LZP03 can reach 4.26 multiplied by 10 10 cfu/mL, then detect fermentation waste liquid after centrifugation collection thalli, it has better clearance to raise pigs waste water pollutant to find it, the organic matter clearance of LZP03 is 89.62%, the COD clearance reaches 92%, the nitrogen removal rate in ammonia state is 90.35%, the phosphorus content clearance can reach 69.10%, and LZP03 can effectively reduce the pH of original waste water of raising pigs, make it reduce to neutral or acid pH solution by original alkaline solution, prevent to filter out waste water and cause soil salinization, destroy soil structure.
Therefore, the technology of realizing the resource utilization of the pig raising wastewater by the beneficial microorganisms has the advantages that the decontamination capability of the pig raising wastewater is not weaker than that of the domestic and foreign mainstream physicochemical treatment technology, natural treatment technology and traditional biological treatment technology, the effect is even better, and the technology has higher broad spectrum for removing pollutants in the pig raising wastewater more comprehensively; the pig raising wastewater treatment by microbial fermentation is lower in decontamination cost, less in equipment investment, short in decontamination period, free of a large amount of fields, and capable of building small fermentation equipment close to the source for pig raising wastewater treatment, and small enterprises can also bear system construction cost; the beneficial microorganisms are used for fermentation, beneficial microorganism strains can be harvested for production and use of microbial agents while the pig raising wastewater is treated, the damage of pollutants to the environment is reduced in the pig raising wastewater treatment process, economic benefits can be reported, the microbial agents can be promoted to be popularized and the development of green agriculture can be accelerated, a virtuous cycle system of ecological protection-agricultural development is created, and the method accords with the slightly large direction of sustainable development war in China.
Drawings
FIG. 1 shows the change of biomass of LZP03 strain in the pig-raising wastewater after the original and optimized carbon-nitrogen ratio with time;
FIG. 2 is a comparison of the change in total nitrogen content and the reduction rate (A) of the pig wastewater (C) treated by LZP03 in the original (B) and the optimized pig wastewater (C);
FIG. 3 is a comparison of the change and decrease rate (A) of the total carbon content of the original (B) and the optimized swine wastewater (C) after the LZP03 treatment;
FIG. 4 is a comparison of the change in total organic carbon content and the reduction rate (A) of the original (B) and the optimized swine wastewater (C) after the LZP03 treatment;
FIG. 5 is a comparison of the reduction rates of total nitrogen, total carbon and total organic carbon in the supernatant of LZP03 fermentation broth;
FIG. 6 is the pH trend of LZP03 in optimized medium as a function of fermentation time;
FIG. 7 shows the removal rate of COD by LZP03 in the fermentation broth and the trend with time;
FIG. 8 shows the removal rate of ammonia nitrogen and phosphorus by LZP 03.
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 screening of strains and their use in wastewater treatment for swine
1 materials and methods
1.1 physical and chemical Properties of original wastewater from pig raising
The pig raising wastewater used in the test is taken from a certain pig farm in the Fulal base area in the Qiqi Harry city, and the collected pig raising wastewater is stored at 4 ℃ for subsequent experiments, wherein the basic characteristics of the pig raising wastewater are shown in Table 1.
TABLE 1 physicochemical properties of pig wastewater
1.2 sources of test strains
The strains used in the test are different kinds of pure culture beneficial microorganisms obtained by separating and screening the subject groups, and the research finds that all strains are high-efficiency strains with better industrial application potential in the aspects of plant growth promotion, biological prevention and control, pollutant prevention and control and the like, the strains are stored in a refrigerator at the temperature of minus 80 ℃ for standby application, all strains are classified and identified by strain morphology and 16SrDNA, and the identification list is shown in Table 2.
TABLE 2 test strains
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 distilled water to a constant volume of 1000.00mL, wherein the agar is not added into a liquid culture medium.
Original pig raising wastewater culture medium: pig raising wastewater transported from a pig farm.
The pig raising wastewater culture medium with the optimized carbon-nitrogen ratio comprises the following steps: 10g of brown sugar, and fixing the volume of the pig raising wastewater to 1000.00mL, so that the carbon-nitrogen ratio of the pig raising wastewater is 16-20:1.
1.4 high-efficiency degrading strain primary screening for pig-raising wastewater
1.4.1 seed liquid preparation
The test strains in the table 2 are respectively inoculated to a beef extract peptone culture medium and cultured for 24h at the temperature of 30 ℃ for activation. The activated strains were inoculated into a beef extract peptone medium, subjected to shaking culture at 30 ℃ and 120r/min, and then each strain cultured to the same concentration (OD 600= 1.0) was used as a seed solution for a treatment test of swine wastewater.
1.4.2 preliminary screening of strains
In order to screen strains capable of growing in large quantities in the pig-raising wastewater, seed solutions of different strains are respectively inoculated into triangular flasks filled with 100mL of sterilized pig-raising wastewater according to the inoculation amount of 1vol%, shake-flask culture is carried out at 30 ℃ at 120r/min, fermentation liquor is sampled every 24h, and the maximum viable bacteria amount of the strains is determined by detecting the viable bacteria amount in each time period. The viable count is determined by a dilution coating method, 10mL of the fermented culture medium is taken and put into a 90mL triangular flask filled with glass beads and sterile water, the flask is shaken for 30min in a shaking table at 200r/min and then is subjected to gradient dilution, 0.1mL of diluent with proper dilution concentration is selected and coated on a solid plate of a beef extract peptone culture medium, the culture is carried out for 24h at 30 ℃, the viable count (cfu/mL) is calculated, and three groups of parallel culture are set. Then selecting the strain with higher fermentation biomass for subsequent experiments.
1.5 research on resource utilization effect of screened strains on pig raising wastewater
1.5.1 changes of growth conditions and treatment effects of various screened strains in raw wastewater and carbon-nitrogen ratio optimization wastewater over time
Because the carbon-nitrogen ratio in the pig wastewater stock solution is small, the growth and reproduction of thalli can be inhibited, and the treatment effect of the pig wastewater is reduced, the proper carbon-nitrogen ratio optimization is carried out on the pig wastewater culture medium, whether the content of the thalli can be improved or not is detected, and the treatment capacity of the pig wastewater is enhanced. And respectively inoculating the screened dominant strains into a beef extract peptone culture medium to prepare seed liquid, respectively inoculating the dominant strains into 300mL of original pig-raising wastewater culture medium and an optimized culture medium added with 1wt% of brown sugar according to the inoculation amount of 2vol%, carrying out shake-flask culture at 30 ℃ at 120r/min, and sampling samples every 24h to measure various indexes.
1.5.2 treatment Effect of pig raising wastewater with optimized carbon-nitrogen ratio after centrifugation by strain screening treatment
In order to simulate whether the pig raising waste liquid treated by each strain can further reduce the potential of environmental threat after thalli are obtained in the industrial production process, the experiment carries out index detection after sample liquid at different fermentation time is centrifuged. Inoculating the seed solution of the screened strains into an optimized culture medium, carrying out shake flask culture at the temperature of 30 ℃ and at the speed of 120r/min, sampling samples every 24h, centrifuging for 5min under the condition of 12000rpm, and then carrying out index determination.
1.6 test measurement index
1.6.1 Water quality index
And (3) measuring COD, pH, total nitrogen, total carbon, total organic carbon, ammoniacal nitrogen and phosphorus content of the sample.
1.6.2 microbiological indicators
Placing 10mL of fermentation liquor into 90mL of sterile water in a shaking table at 200r/min for shaking for 30min, and then carrying out 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 culture medium plate, culturing at 30 ℃ for 24 hours, calculating the viable count (cfu/mL), and setting three groups of parallels.
2. Results and discussion
2.1 screening of wastewater treatment strains for pig raising
The purpose of screening different strains is achieved by detecting the maximum content of viable bacteria of each strain in the pig-raising wastewater, and the purpose of screening different strains is achieved by culturing beneficial strains by using the pig-raising wastewater, so that the LZP03 can grow in the pig-raising wastewater and has higher biomass, and the maximum value of the viable bacteria number of the LZP03 in the pig-raising wastewater is LZP03=1.76 × 10 10 The cfu/mL was used for subsequent experiments with higher viable bacterial count compared to other strains.
2.2LZP03 comparison of growth of bacteria in original and optimized wastewater culture media for pig raising over time
In order to determine the biomass of the LZP03 strain in different culture media, the strain is respectively inoculated in the original culture medium and the optimized culture medium to carry out viable count determination by a gradient dilution plating method. As can be seen from FIG. 1, when LZP03 grows in the original swine wastewater culture medium, the maximum viable count reaches 1.76X 10 after 72h of culture 10 cfu/mL, the maximum viable count is 4.26 multiplied by 10 after 96 hours in the optimized pig-raising wastewater culture medium 10 cfu/mL, 56.86% biomass is improved, so that the biomass of each strain in the pig raising wastewater can be obviously improved by adding brown sugar for carbon-nitrogen ratio optimization, the treatment capacity of the thalli on the pig raising wastewater is improved, and the thalli acquisition of beneficial microorganisms can be better improved.
2.3 Change of Total Nitrogen, total carbon and Total organic carbon of fermentation broth during growth of LZP03 in original and optimized wastewater culture Medium for pig raising
2.3.1 Change in Total Nitrogen in original and optimized wastewater Medium from pig Breeding
Respectively inoculating the LZP03 to an original pig raising wastewater culture medium and an optimized pig raising wastewater culture medium, and measuring the total nitrogen of the fermentation liquor every 24 hours. As can be seen from FIG. 2, the total nitrogen content of the fermentation broth after the original swine wastewater is inoculated with the LZP03 strain generally shows a decreasing trend along with the increase of the fermentation time of the strain, the total nitrogen content of the LZP03 fermentation broth is decreased to a minimum value of 178.4mg/L when the strain is cultured for 96 hours, and the total nitrogen content is decreased by 55.75 percent compared with the control group. The LZP03 fermentation liquor using the optimized pig raising wastewater culture medium also shows a general reduction trend along with the extension of the fermentation time, the total nitrogen content rapidly decreases in a fermentation range of 0-24h and then gradually decreases, the minimum value of the total nitrogen content is 104.8mg/L respectively when the LZP03 is cultured for 96h, and the total nitrogen content is reduced by 68.62% compared with that of the LZP03 in a control group. The comparison shows that the reduction rate and the reduction speed of the total nitrogen of the fermentation liquor of the LZP03 when the original pig-raising wastewater grows are both smaller than those of the fermentation liquor of the pig-raising wastewater after inoculation of the optimized carbon-nitrogen ratio, and the nitrogen in the culture medium possibly overflows due to denitrification, so that the utilization of the strains on the nitrogen can be better improved after the carbon-nitrogen ratio is optimized.
2.3.2 comparison of Total carbon treatment in the original and optimized wastewater Medium for pig Breeding
And respectively inoculating the screened strains to an original pig raising wastewater culture medium and an optimized pig raising wastewater culture medium, and performing total carbon determination on the fermentation liquor every 24 hours. As shown in figure 3, after the LZP03 strain is inoculated in the original pig-raising wastewater culture medium, the total carbon content in the fermentation liquor is in a trend of slowly decreasing and then tending to be stable along with the increase of the culture time, the minimum value of the total carbon content of the LZP03 fermentation liquor is 740.0mg/L, and the total carbon decrease rate of the screening strain LZP03 fermentation liquor is 24.61% compared with the control group. After the screened strains are inoculated in the optimized pig raising wastewater culture medium, the total carbon content in fermentation liquor of the strains continuously decreases along with the increase of the culture time, the speed is higher, the total carbon content finally tends to slowly decrease, the total carbon content of the fermentation liquor of each strain reaches the minimum LZP03=1722mg/L after 96h of culture, and the total carbon decrease rate is reduced by 66.60 percent compared with that of a control group. The LZP03 is cultured in the optimized culture medium, the total carbon content reduction rate in the fermentation liquor is obviously improved compared with the total carbon reduction rate in the original pig-raising wastewater, 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, so that the growth and metabolism conditions of the microorganisms in the optimized culture medium are more active, and the optimized pig-raising wastewater culture medium can well promote the growth of strains and improve the metabolic capability of the strains.
2.3.3 comparison of the treatment conditions of Total organic carbon in the original and optimized wastewater fermentation broths from pig farming
As can be seen from fig. 4, after inoculation of the LZP03 strain, the total organic carbon content in the original swine wastewater fermentation broth tends to increase first and then decrease with the increase of the culture time of the bacteria, the minimum value content of the total organic carbon after fermentation of the LZP03 is 294.8mg/L compared to the control group, and the decrease rate of the total organic carbon is 18.54%, whereas when the LZP03 is inoculated into the optimized swine wastewater culture medium, the total organic carbon content of the fermentation broth continuously decreases with the increase of the culture time and reaches the minimum value at 96 hours, and the decrease rate of the total organic carbon in the optimized swine wastewater culture medium is 70.56% compared to the control group, and the consumption rate and consumption rate of the total organic carbon in the optimized swine wastewater culture medium are faster compared to the original swine wastewater culture medium. The fact that the total organic carbon in the fermentation liquor of the original pig raising wastewater culture strain is reduced after being increased is probably that the microorganism degrades some insoluble solid pig manure and impurities in the pig raising wastewater, the degradation speed of the thalli is higher than the total organic carbon consumed by the thalli for self synthesis or metabolism in the early period, so that the total organic carbon is continuously accumulated, then the consumption of the total organic carbon is continuously improved along with the continuous increase of the number of the strains, the amount of the added substrate is continuously consumed, so that the accumulation speed of the total organic carbon in the fermentation liquor is lower than the consumption speed, and the content of the total organic carbon is continuously reduced. The continuous reduction of the total organic carbon consumption of the optimized culture medium is probably because the total organic carbon content of thalli rapidly increased in the early stage after carbon source supplement and used for self synthesis and metabolism consumption is far greater than the accumulated amount, so the total organic carbon content is continuously reduced.
2.4 change of indexes of fermentation waste liquid after removing bacteria from swine waste water treated by LZP03
Through comparison of changes of biomass, total nitrogen, total carbon and total organic carbon of the LZP03 in the growth process of the original and optimized pig raising wastewater culture media, the strain can better promote growth and reproduction biomass in the optimized pig raising wastewater culture media, and has stronger metabolism and utilization capacity for nutrients in the pig raising wastewater, so that the optimized pig raising wastewater culture media are selected as basic culture media for beneficial microbial industrial fermentation, and in order to simulate whether the LZP03 can better reduce potential threat of filtered fermentation waste liquid to the environment after fermenting in the pig raising wastewater and harvesting bacteria in the industrial production process, the experiment removes the bacteria in the strain fermentation liquid by a high-speed centrifugation method and determines the change conditions of the total nitrogen, the total carbon, the total organic carbon, COD, ammonia nitrogen and phosphorus in a supernatant.
2.4.1 changes in Total Nitrogen, total carbon and Total organic carbon content of LZP03 in the optimized wastewater culture Medium for pig raising
And inoculating LZP03 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. 5, compared with the control group, the total nitrogen content in the centrifuged supernatant of the LZP03 fermentation liquid is reduced by 94.58%, the total carbon content is reduced by 85.96%, and the total organic carbon content is reduced by 89.62%, which indicates that the LZP03 strain can better utilize the nutrient components in the swine wastewater to perform self growth, reproduction and metabolic activities, and prevent the filtered fermentation waste liquid from being enriched to cause environmental pollution.
2.4.2 pH Change of the 2.2LZP03 in the optimized wastewater culture Medium for pig farming
The pH of the swine wastewater is reduced from the original alkaline solution with pH =8.85 to the neutral fermentation broth with pH =7.34 after the swine wastewater is fermented for 96 hours by the LZP03 strain as shown in fig. 6, which is probably because the strain secretes a certain amount of organic acid in the alkaline swine wastewater during the fermentation process by utilizing the nutrients of the swine wastewater to perform self-growth, thereby reducing the pH of the high alkaline swine wastewater, which is of great significance for preventing the filtered wastewater from causing soil salinization and damaging the soil structure.
2.4.3 change of COD content and removal rate of LZP03 in optimized pig-raising wastewater culture medium
In order to determine the effect of screening the strain on removing COD from the pig wastewater, COD detection is carried out on the supernatant of the LZP03 fermentation liquor, and as can be seen from figure 7, along with the continuous increase of the fermentation time of the strain, the COD content of the strain is continuously and rapidly reduced from original COD =16000mg/L and is maximally reduced to COD =1280mg/L, and the COD removal rate reaches LZP03=92%, so that the LZP03 has a better removal rate on the COD, the threat of the filtered waste liquor on the environment is reduced, and the COD content of the strain is found to be slightly increased in a time interval of culturing 72h-96h in a trend graph of the change of the LZP03 fermentation liquor along with the time, and probably because the newly increased number of thalli cells at the end of the strain entering the stable period is smaller than the reduction capability of removing the COD and the COD concentration is increased to some extent due to the outflow of substances of dead and broken cells.
2.4.4LZP03 removal rate of ammoniacal nitrogen and phosphorus in optimized pig raising wastewater
The ammonia nitrogen content of the fermentation liquor supernatant of the LZP03 at different fermentation times is measured, and as can be seen from figure 8, the LZP03 strain can greatly reduce the ammonia nitrogen content of the fermentation waste liquor, the removal rate of the ammonia nitrogen reaches 90.35%, and the removal effect of the ammonia nitrogen is good. 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. It can be seen from the figure that the phosphorus removal rate of the sample can reach 69.10% compared with that of the control LZP03, indicating that the sample has a certain phosphorus removal capability.
3 conclusion
(1) The LZP03 can grow in the original pig raising wastewater and has higher biomass, and the biomass is improved more obviously in the pig raising wastewater with optimized carbon-nitrogen ratio, wherein the maximum viable count of the LZP03 is 4.26 multiplied by 10 10 cfu/mL, and through comparing the situation that the LZP03 strain utilizes carbon, nitrogen and organic carbon in the original and optimized pig-raising wastewater, the metabolism of each strain in the optimized culture medium is more vigorous, so that the optimized carbon and nitrogen is more suitable for the growth and reproduction of the strain and the resource utilization of the optimized pig-raising wastewater.
(2) The LZP03 strain is fermented in the pig raising wastewater culture medium with the optimized carbon-nitrogen ratio, and has better removal rate on COD, ammonia nitrogen and phosphorus content, the COD removal rate reaches 92%, the ammonia nitrogen removal rate is 90.35%, the phosphorus removal rate is 69.10%, the pH value of the original pig raising wastewater can be effectively adjusted, the possibility of fermentation waste liquid on environmental pollution is reduced, and the resource utilization of the pig raising wastewater is better improved.
Claims (10)
1. The application of the Bacillus megaterium LZP03 in treating the pig wastewater is characterized in that the Bacillus megaterium LZP03 is preserved in the China center for type culture collection, and the strain preservation number is CCTCC NO. M2018599.
2. The use of claim 1, wherein the seed solution of Bacillus megaterium LZP03 is inoculated into sterilized swine wastewater to be treated, and the swine wastewater is fermented to obtain treated swine wastewater, wherein the treated swine wastewater has a reduced pollutant content compared to the swine wastewater before the treatment.
3. The use of claim 1, further comprising the step of adding brown sugar to the sterilized swine wastewater to be treated so that the swine wastewater has a carbon-to-nitrogen ratio of 16-20:1..
4. The use of claim 2, wherein the fermentation is carried out at 30 ℃ and 120r/min for 24-96h.
5. The use according to claim 3, wherein the pig wastewater after treatment has a reduced content of total nitrogen, total carbon, total organic carbon, COD, ammoniacal nitrogen, phosphorus compared to the pig wastewater before treatment.
6. A method for treating pig raising wastewater is characterized by comprising the following steps:
(1) Inoculating Bacillus megaterium LZP03 into beef extract peptone culture medium for activation, inoculating activated strain into beef extract peptone culture medium, and regulating strain concentration to OD with sterile water 600 =1.0 as seedLiquid; the bacillus megatherium LZP03 is preserved in China center for type culture collection, and the strain preservation number is CCTCC NO. M2018599;
(2) Inoculating the seed solution into sterilized wastewater to be treated for pig raising, and fermenting to obtain the treated wastewater for pig raising, wherein the content of pollutants in the treated wastewater for pig raising is reduced compared with that before treatment.
7. The method as claimed in claim 6, wherein the step (2) further comprises the step of adding brown sugar to the sterilized swine wastewater to be treated so that the carbon-nitrogen ratio of the swine wastewater is 16-20:1.
8. the method according to claim 6, wherein the step (2) comprises inoculating the sterilized waste water for swine cultivation with the seed solution in an amount of 1 to 2 vol%.
9. The method of claim 6, wherein the fermentation in step (2) is performed at 30 ℃ and 120r/min for 24-96h.
10. The method of claim 6, wherein the pig wastewater after treatment has a reduced content of total nitrogen, total carbon, total organic carbon, COD, ammoniacal nitrogen, phosphorus compared to the pig wastewater before treatment.
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