CN114606154A - Tail vegetable wastewater recycling microbial inoculum and application thereof in preparation of plant nutrient solution - Google Patents

Abstract

The invention discloses a waste water recycling microbial inoculum HQY-of-002 prepared from Bacillus licheniformis (Bacillus licheniformis)Bacillus licheniformis) ACCC 01050, Bacillus amyloliquefaciens (A)Bacillus amyloliquefaciens) CCTCC NO: m2017040, vegetable bacillus (B.plantarum) (B.plantarum)Bacillus oleronius) CCTCC NO: m2017039 and Pichia pastoris (Pichia farinose) CCTCC NO: m2017044. The strain in the microbial agent disclosed by the invention is selected from strains which are not used for toxicological tests in general technical guidelines for biological safety of microbial fertilizers (NY 1109-2006) and strains which are issued by Ministry of health and informed by List of strains which can be used for food, does not contain heavy metals and toxic and harmful chemical substances, and solid products are non-toxic and non-irritant. The waste water recycling microbial inoculum of the invention can decompose polysaccharide, protein, fiber and other substances in the waste water of the kale, can ferment the waste water of the kale into plant nutrient solution, and the fermented plant nutrient solution has no external chemical medicine, is rich in a large amount of trace elements required by plant growth, and has a promoting effect on the plant growth.

Description

Recycling inoculant of tail vegetable wastewater and its application in the preparation of plant nutrient solution

technical field

The invention relates to a cauliflower waste water resource-recycling inoculant and its application in microbial treatment of plant waste water to prepare plant nutrient solution.

Background technique

my country is a big country producing cauliflower waste. The water content of cauliflower waste is as high as 90%. In the process of realizing harmlessness, reduction and resource utilization of cauliflower waste, a large amount of high-concentration wastewater will inevitably be generated, that is, cauliflower waste. waste water. The COD in the wastewater produced by tail vegetables is as high as 30,000mg/L, the ammonia nitrogen is as high as 500mg/L, and the total phosphorus is as high as 120mg/L. If it is treated as sewage, whether it meets the surface water discharge standard or the management standard, it will face huge treatment pressure.

Microorganisms, as natural decomposers, can decompose the residues of cauliflower into components that can be absorbed by plants and return them to the soil. Therefore, if the bacteria that can decompose polysaccharides, proteins, fibers and other substances in cauliflower are used to convert them into substances that can be absorbed by plants , which will greatly reduce the pressure on the environment, and turn waste into treasure, which is of great significance to the vegetable industry.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide a cauliflower waste water resource-recycling inoculant, and its application in the preparation of plant nutrient solution in the treatment of plant waste water by resource-recycling microorganisms.

The technical scheme adopted in the present invention is:

HQY-of-002, a cauliflower wastewater resource-recycling agent, is produced by Bacillus licheniformis ACCC 01050, Bacillus amyloliquefaciens CCTCC NO: M2017040, and vegetable Bacillus oleronius CCTCC NO: M2017039 and Pichia farinose CCTCC NO: M2017044.

The strains in the microbial inoculum of the present invention are selected from the strains exempted from toxicological tests in the "General Technical Guidelines for Biosafety of Microbial Fertilizers (NY1109-2006)" and in the notice issued by the Ministry of Health on the "List of strains that can be used in food" It does not contain heavy metals and toxic chemicals, and the physical product is non-toxic and non-irritating. Practical application will not cause secondary pollution to the environment, and will not cause additional burdens. It belongs to an environment-friendly and affinity microbial inoculum product, and the inoculum is simple and efficient in storage, transportation and delivery. The process is simple to operate, no external chemicals are added, and the fermented plant nutrient solution has no external chemicals added, is rich in macro and trace elements required for plant growth, and has a promoting effect on plant growth.

Preferably, in terms of the proportion of viable bacteria cultured to the middle and late logarithmic stage, the composition is as follows: Bacillus licheniformis ( Bacillus licheniformis ) ACCC 01050 25~35%, Bacillus amyloliquefaciens ( Bacillus amyloliquefaciens ) CCTCC NO: M2017040 25%~ 35%, Bacillus oleronius CCTCC NO: M2017039 20%~30%, Pichia farinose CCTCC NO: M2017044 15%~25%.

More preferably, the composition of the tail vegetable wastewater resource-recycling inoculant is as follows: Bacillus licheniformis ACCC 01050 30%, Bacillus amyloliquefaciens CCTCCNO: M2017040 25%, Bacillus oleronius CCTCC NO: M2017039 25%, Pichia farinose CCTCC NO: M2017044 20%.

The preparation method of HQY-of-002 for cauliflower wastewater resource-recycling in the present invention is as follows: inserting various strains into a liquid strain activation medium, and aerobic activation at 30° C. to 36° C. for 10 hours to 16 hours to prepare a seed activation solution, Then, the inoculation amount of 1%~3% by volume is inserted into the seed medium, and aerobic fermentation at 30℃~36℃ is made for 12h~18h to make seed liquid. The inoculum amount of 5% by volume was inserted into the bacterial fermentation medium, and aerobic fermentation was carried out at 30 ℃ ~ 35 ℃ for 18 h ~ 24 h to prepare the tail vegetable wastewater resource recycling bacterial agent HQY-of-002.

The composition of the bacterial activation medium of tail vegetable wastewater resource-recycling bacterial agent HQY-of-002 is as follows: 0.8%~1.2% (w/v, 1% means that 100mL medium contains 1g of this substance, the same below) peptone, 0.4% ~0.6% yeast extract, 1.8%~2.2% glucose, 0.8~1.2% sodium chloride, 1.8‰~2.2‰ dipotassium hydrogen phosphate, 0.8‰~1.2‰ ammonium chloride, 0.18‰~0.22‰ magnesium sulfate heptahydrate, 0.048‰~0.052‰ manganese sulfate, pH adjusted to about 7.0, sterilized at 115℃ for 30 minutes. The composition of the seed medium is as follows: 1.8%~2.2% peptone, 0.8%~1.2% glucose, 0.8%~1.2% molasses, 0.8~1.2% sodium chloride, 1.8‰~2.2‰ dipotassium hydrogen phosphate, 1.8‰~2.2‰ Ammonium chloride, 0.18‰~0.22‰ magnesium sulfate heptahydrate, 0.048‰~0.052‰ manganese sulfate, pH adjusted to about 7.0, sterilized at 115℃ for 30 minutes. The composition of fermentation medium is characterized by: 60%~70% tail vegetable wastewater, 0.8~1.2% molasses, 0.4~0.6% peptone, 0.5~0.8% sodium chloride, 1.8‰~2.2‰ dipotassium hydrogen phosphate, 0.18‰~0.22 ‰ Magnesium sulfate heptahydrate, adjust the pH to about 7.0, and sterilize at 115°C for 30 minutes.

The invention also relates to the application of the cauliflower waste water resource-recycling inoculant in the preparation of plant nutrient solution by resource-recycling microorganisms for treating plant waste water.

Specifically, the plant wastewater is plant wastewater other than bactericidal plants.

Specifically, the application is: adding the tail vegetable wastewater resource-recycling bacterial agent HQY-of-002 to the plant wastewater, performing aerobic fermentation at 30 ° C ~ 37 ° C, and controlling the dissolved oxygen to 2 ~ 4 mg/L, Fermentation is carried out for 3-6 days, and after the fermentation is completed, it is diluted to COD 100-1000 mg/L, which is the plant nutrient solution.

Preferably, the aerobic fermentation is carried out at 33°C to 36°C.

The beneficial effects of the invention are mainly reflected in: the tail vegetable wastewater resource-recycling inoculant can decompose the polysaccharide, protein, fiber and other substances in the tail vegetable, and can ferment the tail vegetable waste water into a plant nutrient solution, and the fermented plant nutrient solution has no external The addition of chemicals is rich in macro and trace elements required for plant growth, and has a promoting effect on plant growth.

Description of drawings

Fig. 1 is the 4th day photo of embodiment 4 seed germination experiments;

Fig. 2 is the contrast of seed germination index on the 5th day of embodiment 4;

Fig. 3 is a photograph on the 5th day of the seed germination experiment of Example 4.

(5) Specific implementation methods

In order to deepen the understanding of the present invention, the present invention will be described in further detail below with reference to specific embodiments, which are only used to explain the present invention and do not limit the protection scope of the present invention.

Embodiment 1: Preparation of tail vegetable waste water resource recycling bacterial agent HQY-of-002

The inoculum is prepared through the following steps:

(1) Take out the various strains stored in the glycerol cryovials in the -80°C refrigerator, thaw them in the ultra-clean workbench sterilized by ultraviolet rays, and aseptically suck 1 ml of the pipette into the sterilized 10 ml liquid strains for activation Activated in the medium.

(2) The liquid strain activation medium used in the preparation step (1) of this inoculum is prepared as follows: 1.0% peptone, 0.5% yeast extract, 2.0% glucose, 1.0% sodium chloride, 2.0‰ dipotassium hydrogen phosphate, 1.0‰ ammonium chloride, 0.2‰ magnesium sulfate heptahydrate, 0.05‰ manganese sulfate, pH adjusted to about 7.0. The medium was sterilized at 115°C for 30 minutes.

(3) In the preparation step (1) of this bacterial agent, after the bacteria are connected to the activation medium, the activation solution can be prepared by aerobic activation at 30 ℃ ~ 36 ℃ for 10h ~ 16h.

(4) Under aseptic conditions, absorb the activated strain activation solution in (3), insert it into the seed medium with 3% inoculum, and ferment it aerobic at 30°C to 36°C for 12 hours to make a seed solution.

(5) The preparation method of the seed medium used in step (4) is: 2.0% peptone, 1.0% glucose, 1.0% molasses, 1.0% sodium chloride, 2.0‰ dipotassium hydrogen phosphate, 2.0‰ ammonium chloride, 0.2‰ seven Water magnesium sulfate, 0.05‰ manganese sulfate, pH adjusted to about 7.0. The medium was sterilized at 115°C for 30 minutes.

(6) Under aseptic conditions, absorb the seed liquid that has been fermented in (4), and the seed liquid is in accordance with Bacillus licheniformis ACCC 01050 30%, Bacillus amyloliquefaciens CCTCC NO: M2017040 25%, vegetable spores Bacillus oleronius CCTCC NO: M2017039 25%, Pichia farinose CCTCC NO: M2017044 20% were mixed in a proportion of 5% inoculum into the bacterial fermentation medium, 30 ℃ ~ 35 ℃ aerobic fermentation The bacterial agent HQY-of-002 can be made in 24 hours.

(7) The preparation method of the bacterial fermentation medium used in step (6) is: 70% tail vegetable wastewater, 1.0% molasses, 0.5% peptone, 0.6% sodium chloride, 2.0‰ dipotassium hydrogen phosphate, 0.2‰ heptahydrate magnesium sulfate, and the pH was adjusted to about 7.0, and the medium was sterilized at 115°C for 30 minutes.

(8) The inoculum HQY-of-002 fermented in step (6), the number of viable bacteria >10 ⁸ CFU/mL, can be stored stably for 6 months at room temperature, and the best application temperature: 25~45℃; pH : 4~8.5.

Embodiment 2: Preparation of tail vegetable waste water resource recycling bacterial agent HQY-of-003

The inoculum is prepared through the following steps:

(1) Take out the various strains stored in the glycerol cryovials in the -80°C refrigerator, thaw them in the ultra-clean workbench sterilized by ultraviolet rays, and aseptically suck 1 ml of the pipette into the sterilized 10 ml liquid strains for activation Activated in the medium.

(2) The liquid strain activation medium used in the preparation step (1) of this inoculum is prepared as follows: 1.0% peptone, 0.5% yeast extract, 2.0% glucose, 1.0% sodium chloride, 2.0‰ dipotassium hydrogen phosphate, 1.0‰ ammonium chloride, 0.2‰ magnesium sulfate heptahydrate, 0.05‰ manganese sulfate, pH adjusted to about 7.0. The medium was sterilized at 115°C for 30 minutes.

(3) In the preparation step (1) of this bacterial agent, after the bacteria are connected to the activation medium, the activation solution can be prepared by aerobic activation at 30 ℃ ~ 36 ℃ for 10h ~ 16h.

(4) Under aseptic conditions, absorb the activated strain activation solution in (3), insert it into the seed medium with 3% inoculum, and ferment it aerobic at 30°C to 36°C for 12 hours to make a seed solution.

(5) The preparation method of the seed medium used in step (4) is: 2.0% peptone, 1.0% glucose, 1.0% molasses, 1.0% sodium chloride, 2.0‰ dipotassium hydrogen phosphate, 2.0‰ ammonium chloride, 0.2‰ seven Water magnesium sulfate, 0.05‰ manganese sulfate, pH adjusted to about 7.0. The medium was sterilized at 115°C for 30 minutes.

(6) Under sterile conditions, suck the seed liquid that has been fermented in (4), and the seed liquid is according to Bacillus licheniformis ACCC 01050 30%, Bacillus amyloliquefaciens CCTCC NO: M2017040 40%, Pichia Yeast ( Pichia farinose ) CCTCC NO: M2017044 30%. After mixing, 5% of the inoculum was added to the bacterial fermentation medium, and the bacterial agent HQY-of-003 could be prepared by aerobic fermentation at 30°C to 35°C for 24 hours.

(7) The preparation method of the bacterial fermentation medium used in step (6) is: 70% tail vegetable wastewater, 1.0% molasses, 0.5% peptone, 0.6% sodium chloride, 2.0‰ dipotassium hydrogen phosphate, 0.2‰ heptahydrate magnesium sulfate, and the pH was adjusted to about 7.0, and the medium was sterilized at 115°C for 30 minutes.

(8) The bacterial agent HQY-of-003 fermented in step (6), the number of viable bacteria is more than 10 ⁸ CFU/mL, it can be stored stably for 6 months at room temperature, and the best application temperature is 25~45℃; pH : 4~8.5.

Example 3: Preparation of tail vegetable wastewater resource-recycling bacterial agent HQY-of-004

The inoculum is prepared through the following steps:

(1) Take out the various strains stored in the glycerol cryovials in the -80°C refrigerator, thaw them in the ultra-clean workbench sterilized by ultraviolet rays, and aseptically suck 1 ml of the pipette into the sterilized 10 ml liquid strains for activation Activated in the medium.

(2) The liquid strain activation medium used in the preparation step (1) of this inoculum is prepared as follows: 1.0% peptone, 0.5% yeast extract, 2.0% glucose, 1.0% sodium chloride, 2.0‰ dipotassium hydrogen phosphate, 1.0‰ ammonium chloride, 0.2‰ magnesium sulfate heptahydrate, 0.05‰ manganese sulfate, pH adjusted to about 7.0. The medium was sterilized at 115°C for 30 minutes.

(3) In the preparation step (1) of this bacterial agent, after the bacteria are connected to the activation medium, the activation solution can be prepared by aerobic activation at 30 ℃ ~ 36 ℃ for 10h ~ 16h.

(4) Under aseptic conditions, absorb the activated strain activation solution in (3), insert it into the seed medium with 3% inoculum, and ferment it aerobic at 30°C to 36°C for 12 hours to make a seed solution.

(5) The preparation method of the seed medium used in step (4) is: 2.0% peptone, 1.0% glucose, 1.0% molasses, 1.0% sodium chloride, 2.0‰ dipotassium hydrogen phosphate, 2.0‰ ammonium chloride, 0.2‰ seven Water magnesium sulfate, 0.05‰ manganese sulfate, pH adjusted to about 7.0. The medium was sterilized at 115°C for 30 minutes.

( 6 ) Absorb the seed liquid that has been fermented in (4) under aseptic conditions. Pichia farinose CCTCC NO: M201704430%. After mixing, 5% of the inoculum was added to the bacterial fermentation medium, and the bacterial agent HQY-of-004 could be prepared by aerobic fermentation at 30℃~35℃ for 24h.

(7) The preparation method of the bacterial fermentation medium used in step (6) is: 70% tail vegetable wastewater, 1.0% molasses, 0.5% peptone, 0.6% sodium chloride, 2.0‰ dipotassium hydrogen phosphate, 0.2‰ heptahydrate magnesium sulfate, and the pH was adjusted to about 7.0, and the medium was sterilized at 115°C for 30 minutes.

(8) The bacterial agent HQY-of-004 fermented in step (6), the number of viable bacteria is more than 10 ⁸ CFU/mL, it can be stored stably for 6 months at room temperature, and the best application temperature: 25~45℃; pH : 4~8.5.

Example 4:

This process is used to treat the tail vegetables produced in the vegetable warehouse: mixed tail vegetables such as cabbage, tomatoes, beans, cabbage, potatoes, carrots, etc. (the COD value of the wastewater is 28500mg/L, the ammonia nitrogen content is 431mg/L, and the total phosphorus is 119mg/L. ), add 3‰ bacterial agent HQY-of-002 to the wastewater, 35 ℃, pH: 5~8, the dissolved oxygen is controlled at 2mg/L~4mg/L under the condition of aerobic fermentation for 6 days, after the fermentation is completed, the pure water is diluted When the COD reaches about 100mg/L, it is the plant nutrient solution.

Four-season fast food (small cabbage) seeds from Jiaxing Jiafeng Seed Co., Ltd., Zhejiang Province were used, and the seeds with full grains were selected for testing. The test method refers to the seed germination index in the agricultural industry standard "Technical Regulations for the Harmless Treatment of Vegetable Wastes at High Temperature Composting (NY/T3441-2019)", and the germination method test is performed. Draw 10ml of nutrient solution into a 9cm petri dish covered with filter paper, put 50 seeds into each dish evenly, then put it into a 30°C incubator, germinate in the dark for 48h, and count the germination rate and root length. Three replicates were made for each sample, and distilled water was used as the control group.

On the 4th day of germination, the germination rate of the seeds reached 100% (see Figure 1), and the seed germination index reached 126% on the 5th day of germination (see Figures 2~3).

Example 5:

This process is used to treat the tail vegetables produced in the vegetable warehouse: mixed tail vegetables such as cabbage, tomatoes, beans, cabbage, potatoes, carrots, etc. (the COD value of the wastewater is 28500mg/L, the ammonia nitrogen content is 431mg/L, and the total phosphorus is 119mg/L. ), add 3‰ bacterial agent HQY-of-003 to the wastewater, 35℃, pH: 5~8, the dissolved oxygen is controlled at 2mg/L~4mg/L for aerobic fermentation for 6 days, after the fermentation is completed, the pure water is diluted When the COD reaches about 100mg/L, it is the plant nutrient solution.

Four-season fast food (small cabbage) seeds from Jiaxing Jiafeng Seed Co., Ltd., Zhejiang Province were used, and the seeds with full grains were selected for testing. The test method refers to the seed germination index in the agricultural industry standard "Technical Regulations for the Harmless Treatment of Vegetable Wastes at High Temperature Composting (NY/T3441-2019)", and the germination method test is performed. Draw 10ml of nutrient solution into a 9cm petri dish covered with filter paper, put 50 seeds into each dish evenly, then put it into a 30°C incubator, germinate in the dark for 48h, and count the germination rate and root length. Three replicates were made for each sample, and distilled water was used as the control group.

The measured seed germination rate reached 84%, and the seed germination index reached 92%.

Example 6:

This process is used to treat the tail vegetables produced in the vegetable warehouse: mixed tail vegetables such as cabbage, tomatoes, beans, cabbage, potatoes, carrots, etc. (the COD value of the wastewater is 28500mg/L, the ammonia nitrogen content is 431mg/L, and the total phosphorus is 119mg/L. ), add 3‰ bacterial agent HQY-of-004 to the wastewater, 35℃, pH: 5~8, the dissolved oxygen is controlled at 2mg/L~4mg/L for aerobic fermentation for 6 days, diluted with pure water after the fermentation is completed When the COD reaches about 100mg/L, it is the plant nutrient solution.

Four-season fast food (small cabbage) seeds from Jiaxing Jiafeng Seed Co., Ltd., Zhejiang Province were used, and the seeds with full grains were selected for testing. The test method refers to the seed germination index in the agricultural industry standard "Technical Regulations for the Harmless Treatment of Vegetable Wastes at High Temperature Composting (NY/T3441-2019)", and the germination method test is performed. Draw 10ml of nutrient solution into a 9cm petri dish covered with filter paper, put 50 seeds into each dish evenly, then put it into a 30°C incubator, germinate in the dark for 48h, and count the germination rate and root length. Three replicates were made for each sample, and distilled water was used as the control group.

The measured seed germination rate reached 78%, and the seed germination index reached 80%.

Example 7:

This process is used to treat tail vegetable and plant waste water: mixed waste water such as baby cabbage, baby cabbage, cucumber, tomato, beans, cabbage, potatoes, carrots, aquatic ornamental plants, etc. , total phosphorus 104mg/L), add 4‰ bacterial agent HQY-of-002 in wastewater, 35℃, pH: 5~8, dissolved oxygen is controlled at 2mg/L~4mg/L under the condition of aerobic fermentation for 6 days, After the fermentation is completed, the pure water is diluted to a COD of about 1000 mg/L, which is the plant nutrient solution.

Four-season fast food (small cabbage) seeds from Jiaxing Jiafeng Seed Co., Ltd., Zhejiang Province were used, and the seeds with full grains were selected for testing. The test method refers to the seed germination index in the agricultural industry standard "Technical Regulations for the Harmless Treatment of Vegetable Wastes at High Temperature Composting (NY/T3441-2019)", and the germination method test is performed. Draw 10ml of nutrient solution into a 9cm petri dish covered with filter paper, put 50 seeds into each dish evenly, then put it into a 30°C incubator, germinate in the dark for 48h, and count the germination rate and root length. Three replicates were made for each sample, and distilled water was used as the control group.

The measured seed germination rate reached 100%, and the seed germination index reached 142%.

The bacterial agent HQY-of-002 of the invention does not contain pathogenic bacteria, heavy metals and toxic chemical substances, and the solid product is non-toxic and non-irritating. Practical application will not cause secondary pollution to the environment, and will not cause additional burdens. It belongs to an environment-friendly and affinity microbial inoculum product, and the inoculum is simple and efficient in storage, transportation and delivery. The process of the invention is simple to operate, no external chemicals are added, and the fermented plant nutrient solution is rich in macro and trace elements required for plant growth, and has a good promoting effect on plant growth.

The foregoing description has fully disclosed specific embodiments of the present invention. It should be pointed out that any changes made by those skilled in the art to the specific embodiments of the present invention will not depart from the scope of the claims of the present invention. Accordingly, the scope of the claims of the present invention is not limited to the foregoing specific embodiments.

Claims (6)

1. HQY-of-002, a cauliflower wastewater resource-recycling agent, is composed of Bacillus licheniformis ACCC 01050, Bacillus amyloliquefaciens CCTCC NO: M2017040, and vegetable Bacillus oleronius CCTCC NO : M2017039 and Pichia farinose CCTCCNO: M2017044 composition. 2. cauliflower waste water resource utilization microbial inoculum as claimed in claim 1, it is characterised in that in terms of the number of viable bacteria cultivated to the middle and late stages of logarithm, the composition is as follows: Bacillus licheniformis ACCC 01050 25～ 35%, Bacillus amyloliquefaciens CCTCC NO: M2017040 25%~35%, Bacillus oleronius CCTCC NO: M2017039 20%~30%, Pichia farinose CCTCC NO: M2017044 15 %~25%. 3. The cauliflower waste water resource microbial agent as claimed in claim 2, characterized in that, the cauliflower waste water resource microbial agent is composed as follows: Bacillus licheniformis ACCC 01050 30%, Bacillus amyloliquefaciens ( Bacillus licheniformis ) Bacillus amyloliquefaciens ) CCTCC NO: M2017040 25%, vegetable Bacillus ( Bacillus oleronius ) CCTCC NO: M2017039 25%, Pichia farinose ( Pichia farinose ) CCTCC NO: M201704420%. 4. the application of the cauliflower waste water resource-recycling inoculant described in one of claims 1 to 3 in the preparation of plant nutrient solution by recycling microorganisms to treat plant waste water. 5. The application according to claim 4, characterized in that the plant waste water is plant waste water other than bactericidal plants. 6. application as claimed in claim 4, it is characterized in that described application is: in plant waste water, add described tail vegetable waste water resource utilization bacterial agent HQY-of-002, at 30 ℃～37 ℃, carry out aerobic Fermentation, controlled dissolved oxygen 2~4mg/L, fermented for 3~6 days, and diluted to COD 100~1000mg/L after the fermentation was completed, that is, the plant nutrient solution.

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