CN108423838B - Microbial preparation for ecological safety water system and preparation method thereof - Google Patents

Microbial preparation for ecological safety water system and preparation method thereof Download PDF

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CN108423838B
CN108423838B CN201810127446.9A CN201810127446A CN108423838B CN 108423838 B CN108423838 B CN 108423838B CN 201810127446 A CN201810127446 A CN 201810127446A CN 108423838 B CN108423838 B CN 108423838B
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microbial
water
strain
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CN108423838A (en
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苏清海
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Pulege Xiamen Biotechnology Co ltd
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    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00Biological treatment of water, waste water, or sewage
    • C02F3/34Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00Biological treatment of water, waste water, or sewage
    • C02F3/34Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • C02F3/347Use of yeasts or fungi
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00Biological treatment of water, waste water, or sewage
    • C02F3/34Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • C02F3/348Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the way or the form in which the microorganisms are added or dosed
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, 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
    • C12N1/14Fungi; Culture media therefor
    • C12N1/16Yeasts; Culture media therefor
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, 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
    • C12N1/20Bacteria; Culture media therefor
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2103/00Nature of the water, waste water, sewage or sludge to be treated
    • C02F2103/007Contaminated open waterways, rivers, lakes or ponds
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2103/00Nature of the water, waste water, sewage or sludge to be treated
    • C02F2103/20Nature of the water, waste water, sewage or sludge to be treated from animal husbandry

Abstract

The invention provides a microbial preparation for an ecological safety water system and a preparation method thereof, and relates to the technical field of water treatment. It comprises the following composite strains: 10-20% of rhodopseudomonas palustris, 20-30% of powdery pichia pastoris, 20-30% of bacillus amyloliquefaciens, 0-15% of rhodococcus erythropolis, 20-30% of lactobacillus plantarum and 5-15% of rhodospirillum sulphureum; and a carrier for loading the composite strain. The preparation method comprises the following steps: adding the composite strain into the culture solution, and performing fermentation culture to obtain a zymophyte solution; concentrating the zymophyte liquid, adding porous starch, stirring and adsorbing, then adding dextrin and lithium tourmaline micropowder, arranging the activated carbon fiber in the activated carbon fiber for adsorption for 2-3 h, and drying. Various strains in the microbial preparation exert mutual synergistic effect, effectively restore water body, form beneficial micro-ecological environment, and fix the strains on the carrier, effectively improve the purification and restoration effects of the microorganisms.

Description

Microbial preparation for ecological safety water system and preparation method thereof
Technical Field
The invention relates to the technical field of water treatment, and particularly relates to a microbial preparation for an ecological safety water system and a preparation method thereof.
Background
The water environment pollution of China is increasingly serious, the water body is polluted due to the discharge of untreated breeding wastewater and industrial and domestic sewage, eutrophication is serious, blue-green algae outbreak and spoilage microorganisms propagate in a large quantity, the ecological environment of the breeding water body or landscape water body is damaged, and serious loss is caused to the aquaculture industry or natural environment, so that the method has important significance on how to restore the water body and construct the ecological safety water system.
In the prior art, the water body restoration mainly comprises a physical method, a chemical method and a biological method. Both physical methods and chemical methods have the problems of large engineering quantity, high cost, incomplete treatment and the like. The biological method mainly based on microbial remediation has unique effect, and can fundamentally realize water purification by absorbing, degrading and converting pollutants and eutrophic components in the water body through the microbes. However, in the prior art, only a single strain is often adopted for water body treatment, and the water body remediation effect is poor. For example, patent CN02156977.0 discloses a low-temperature nitrifying bacteria agent and its use, relating to six strains of hamburger nitrifying bacteria, Vickers nitrifying bacteria, waxy bacillus, Bacillus licheniformis and Saccharomyces cerevisiae, after these strains are cultured, the strains are prepared into the low-temperature resistant nitrifying bacteria agent according to the same volume proportion of the culture, and the low-temperature resistant nitrifying bacteria agent has the function of removing ultra-low concentration ammonia nitrogen at lower temperature, can be used alone or matched with a corresponding bioreactor, and can be used for removing ammonia nitrogen and COD in mariculture and industrial circulating seawater high-density culture. In the technology, the nitrifying bacteria are only suitable for low-temperature culture environments such as mariculture, the application range is narrow, meanwhile, the nitrifying bacteria are required to be matched with a corresponding bioreactor, the composite bacteria are multiple, the steps are complicated, and the popularization and the application in production are not facilitated.
Disclosure of Invention
The invention aims to provide a microbial preparation for an ecological safety water system, which can effectively restore polluted water.
The invention also aims to provide a preparation method of the microbial preparation for the ecological safety water system, which has simple process and easy operation.
The technical problem to be solved by the invention is realized by adopting the following technical scheme.
The invention provides a microbial preparation for an ecological safety water system, which comprises the following components:
the composite strain comprises, by weight, 10-20% of rhodopseudomonas palustris, 20-30% of powdery pichia pastoris, 20-30% of bacillus amyloliquefaciens, 0-15% of rhodococcus erythropolis, 20-30% of lactobacillus plantarum and 5-15% of exothiorhodospira sp; and a carrier for loading the composite strain.
The invention provides a preparation method of a microbial preparation for an ecological safety water system, which comprises the following steps:
s1, preparing a composite strain: mixing strains according to a ratio to obtain the composite strain, adding the composite strain into a culture solution, and carrying out closed culture at the temperature of 28-32 ℃ for 5-10 days to obtain a zymogen solution;
s2, preparing a carrier: concentrating the zymophyte liquid to obtain concentrated bacteria liquid, adding porous starch, stirring and adsorbing, then adding dextrin and lithium tourmaline micro powder to obtain impregnation liquid, arranging activated carbon fibers in the impregnation liquid, adsorbing for 2-3 h, and drying.
The microbial preparation for the ecological safety water system and the preparation method thereof have the beneficial effects that:
the invention utilizes the mutual synergistic effect of the composite strains, can effectively utilize the excessive C, N, P element in the water environment to convert the excessive C, N, P element into beneficial microorganisms, thereby reducing the BOD and COD level in the water environment, forming beneficial micro-ecological environment, purifying water body, reducing the use of chemical disinfector and promoting the growth of cultured animals. The invention can be applied to the purification of aquaculture water, the construction of an ecological safety water system and the restoration of landscape water.
Secondly, the composite strain is fixed on the porous starch, and dextrin is added to realize the strain fixing and embedding effects, so that the treatment effect of the composite strain can be effectively improved, and the adverse phenomena of thallus loss and the like caused by direct inoculation in a water body are avoided. And secondly, adsorbing the porous starch microspheres loaded with the strains into the activated carbon fiber cloth to obtain the loaded microbial preparation, which is convenient to use and transport. When the activated carbon fiber cloth is applied to a water body, the activated carbon fiber cloth firstly adsorbs toxic and harmful substances in the water body, so that the composite strain is prevented from being excessively lost in the early stage, and the double purification effect on the water body is realized.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
The following is a detailed description of the method for preparing the microbial preparation for an eco-safe water system according to the embodiment of the present invention.
The embodiment of the invention provides a microbial preparation for an ecological safety water system, which comprises the following components:
the composite strain comprises, by weight, 10-20% of rhodopseudomonas palustris, 20-30% of powdery pichia pastoris, 20-30% of bacillus amyloliquefaciens, 0-15% of rhodococcus erythropolis, 20-30% of lactobacillus plantarum and 5-15% of exothiorhodospira sp; and the number of the first and second groups,
and the carrier is used for loading the composite strain.
Further, in a preferred embodiment of the invention, the composite strain comprises 15-20% of rhodopseudomonas palustris, 20-25% of pichia pastoris powder, 25-30% of bacillus amyloliquefaciens, 5-10% of rhodococcus erythropolis, 20-25% of lactobacillus plantarum and 5-10% of rhodospirillum sulphureum.
Rhodopseudomonas palustris belongs to one of photosynthetic bacteria, PSB is short for PSB, is one of the oldest strains on the earth, has rich nutrition of PSB thalli, has high protein content of 65 percent, is rich in various vitamins, coenzymes and other bioactive substances and trace elements, has strong adaptability, can endure high-depth organic wastewater and strong decomposition and conversion capacity, has certain tolerance and decomposition capacity on phenol, cyanogen and other toxic substances, and the like. The powdery Pichia pastoris (Pichia farinose) has high tolerance to ammonia nitrogen, can remove ammonia nitrogen, and particularly has good effect on the emission of ammonia gas in animal manure. Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) is harmless to human and livestock, can generate active substances such as antibiotics, antibacterial protein polypeptides and the like, and has a promoting effect on the growth of animals and plants. Rhodococcus erythropolis (Rhodococcus erythropolis) can degrade hydrocarbon pollutants, desulfurize, etc., and Lactobacillus plantarum (Lactobacillus plantarum) can degrade nitrite, etc., and promote animal growth. The Ectothiorhodospira shaposhnikovii can reduce carbon dioxide by using inorganic substances such as hydrogen sulfide and the like as electron donors, synthesize organic substances and effectively degrade sulfides in sewage. Through the compounding of the strains, a good synergistic effect can be exerted, water body purification and restoration can be effectively carried out, the growth of animals or plants in the water body can be promoted, and the immunity and disease resistance of aquatic animals are enhanced.
Further, in a preferred embodiment of the present invention, the rhodopseudomonas palustris (rhodopseudomonas papulosis) is a strain with a preservation number of ACCC10649, the Pichia farinosa (Pichia farinose) is a strain with a preservation number of ACCC20025, the Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) is a strain with a preservation number of ACCC06476, the Rhodococcus erythropolis (rhodochrous erythropolis) is a strain with a preservation number of ACCC02667, the Lactobacillus plantarum (Lactobacillus plantarum) is a strain with a preservation number of ACCC11118, and the rhodospirillum sulphureus (Ectoiodospira shaposhnikovii) is a strain with a preservation number of ACCC 19809.
Further, the strains are produced according to the following method:
rhodopseudomonas palustris (Rhodopseudomonas paussusensis ACCC10649) purchased from the China agricultural microbial strain preservation and management center is transferred into a conical flask of a photosynthetic bacteria culture medium, and is cultured for 60-84 hours under illumination at 28-32 ℃ and taken out after the culture medium is turbid.
The Pichia pastoris (Pichia pastoris ACCC20025) purchased from the China agricultural microbial strain preservation management center is transferred into a wort agar I culture medium, the culture temperature is 25-30 ℃, the shaking culture is carried out at 180-220rpm for 36-60 hours, and the Pichia pastoris is taken out after the culture medium is turbid.
The bacillus amyloliquefaciens (Bacillus amyloliquefaciens ACCC06476) purchased from the China agricultural microbial strain preservation management center is transferred into a nutrient gravy agar culture medium, is subjected to shaking culture at 180-220rpm at the temperature of 32-38 ℃ for 36-60 hours, and is taken out after the culture medium is turbid.
Rhodococcus erythropolis ACCC02667 purchased from China agricultural microbial culture Collection center is transferred into LB culture medium, cultured for 60-84 hours at 28-32 ℃, and taken out after the culture medium is turbid.
Inoculating Lactobacillus plantarum ACCC11118 purchased from China agricultural microorganism culture Collection into an Erlenmeyer flask of MRS liquid culture medium, culturing at 35-39 deg.C for 48-72 hr, and taking out after the culture medium is turbid.
Rhodospirillum oxysporum (Ectothiorhodospirasahophikovii ACCC19809) purchased from the China center for agricultural microbial culture collection is transferred into a conical flask of photosynthetic bacteria culture medium, and is cultured for 60-84 hours under illumination at 28-32 ℃, and is taken out after the culture medium is turbid.
Taking out the above strains after the culture is finished, and then storing in a refrigerator at 2-4 ℃ for later use.
And (3) mixing the strains according to a ratio to obtain the composite strain, adding the composite strain into a culture solution, and carrying out closed culture at the temperature of 28-32 ℃ for 5-10 days to obtain a zymocyte solution. Wherein, the culture solution includes: molasses and water.
Further, in a preferred embodiment of the present invention, the mass ratio of the composite strain to the culture solution is 1: 80-120 parts.
Further, in a preferred embodiment of the invention, the culture solution comprises 8-12 parts by weight of molasses and 90-96 parts by weight of water.
Further, in a preferred embodiment of the present invention, the culture solution further includes 10 to 15 parts by weight of brown sugar.
In the prior art, when microorganisms are applied, the microorganisms are usually directly inoculated in a water body to be treated, but the microorganisms are easily lost due to environmental factors, competitive action and the like. Particularly in a water body with a complex environment, the strains are easily damaged by toxic substances, the survival rate of the microorganisms is low, and the propagation speed is slow. Because, the effect of microorganism on restoring water is not ideal. Therefore, the invention fixes the strain obtained by the method on the carrier, and the method specifically comprises the following steps:
preparing a carrier: concentrating the fermented bacterial liquid obtained by fermentation to obtain concentrated bacterial liquid, adding porous starch, stirring for adsorption, and adding dextrin and lithium tourmaline micropowder to obtain impregnation liquid. And then arranging the activated carbon fiber in the impregnation liquid for adsorption for 2-3 h, and drying to obtain the supported microbial preparation.
Further, in the preferred embodiment of the invention, the fermented bacterial liquid is concentrated to 1/4-1/5 to obtain concentrated bacterial liquid, and the mass ratio of the concentrated bacterial liquid to the porous starch is 1: 0.5-0.7.
Further, the conditions of stirring and adsorbing are as follows: adsorbing for 0.5-1.5 h under the condition of 80-120 r/min. The porous starch is a hydrolysate obtained by degrading various starches by using hydrolase at a temperature lower than the gelatinization temperature of the starch, has a hollow structure similar to a hornet nest, is provided with a large number of small holes on the surface, is low in stacking density and strong in adsorption capacity, and can fully adsorb microorganisms under the stirring condition. And the composite strain has good protection effect after being adsorbed, can effectively protect the composite strain, and has high propagation speed and good water body restoration effect. In this embodiment, the porous starch may be a commercially available product, or may be prepared according to the prior art (e.g., the method disclosed in CN 1546529A), and will not be described herein.
Further, in a preferred embodiment of the invention, the dextrin comprises beta-cyclodextrin and maltodextrin in a mass ratio of 1: 1-2, and the mass ratio of the concentrated bacterial liquid, the lithium tourmaline micro powder and the dextrin is 1: 0.1-0.2: 0.05-0.08.
After the prepared composite strain is adsorbed on the porous starch, the beta-cyclodextrin and the maltodextrin are added to form a buried layer, so that the composite strain is further protected, and the preservation, the transportation and the like of a microbial preparation are facilitated. And a certain amount of lithium tourmaline micro powder is added, the lithium tourmaline micro powder has permanent electricity property and activation property, negative oxygen ions can be generated, the negative ions and water molecules are combined to form small molecular group water, the oxygen content in the water is increased, and the survival rate of the strains is improved. Meanwhile, the negative ions are beneficial to activating cells, improving the disease resistance of aquatic animals and the like.
Further, in a preferred embodiment of the invention, the mass ratio of the concentrated bacterial liquid to the activated carbon fiber cloth is 1: 0.3-0.5.
Further onThe specific surface area of the activated carbon fiber cloth is 1200-1800 m2(ii)/g, the average pore diameter is 0.1 to 1 μm. The activated carbon fiber cloth has a large number of micropores and extremely high specific surface area, and when the activated carbon fiber cloth is arranged in the impregnation liquid, the activated carbon fiber cloth can adsorb porous starch particles loaded with compound strains in the impregnation process to form a loaded microbial preparation. When the microbial preparation is applied to water, the activated carbon fiber cloth can firstly purify the water, particularly can adsorb heavy metal ions such as lead and chromium, and effectively protect the composite strains. On the other hand, a large number of micropores on the activated carbon fiber cloth can ensure good water absorption and water retention, and effectively promote the propagation of microorganisms. When the microorganisms are propagated in a large quantity, the micropores of the activated carbon fiber cloth can also provide adsorption and fixing points for the microorganisms. And good water body purification and restoration effects are achieved through double fixation and double adsorption. Protecting microbial thallus from the influence of external adverse environment (such as acid, alkali, harmful ions, etc.), and preventing mass transfer and the supply of nutrients and the export of products, and maintaining high activity of strains.
The features and properties of the present invention are described in further detail below with reference to examples.
Example 1
The present example provides a microbial preparation, which is prepared according to the following steps:
(1) production and seed production:
rhodopseudomonas palustris (Rhodopseudomonas paussusensis ACCC10649) purchased from the China agricultural microbial strain preservation management center is transferred into a conical flask of a photosynthetic bacteria culture medium, and is cultured for 72 hours under the illumination at the temperature of 30 ℃ and taken out after the culture medium is turbid.
The powdery Pichia pastoris (Pichia farinoseACCC20025) purchased from the China agricultural microbial strain preservation and management center is transferred into a wort agar-I culture medium, the culture temperature is 28 ℃, the shaking culture is carried out at 200rpm for 48 hours, and the powdery Pichia pastoris is taken out after the culture medium is turbid.
Bacillus amyloliquefaciens (Bacillus amyloliquefaciens ACCC06476) purchased from the China agricultural microbial strain preservation management center is transferred into a nutrient gravy agar culture medium, is subjected to shaking culture at 210rpm at 35 ℃ for 48 hours, and is taken out after the culture medium is turbid.
Rhodococcus erythropolis ACCC02667 purchased from China agricultural microbial culture Collection was transferred into LB medium, cultured at 30 ℃ for 72 hours, and taken out when the medium became turbid.
Lactobacillus plantarum ACCC11118 purchased from the China agricultural microbial culture Collection center is transferred into an Erlenmeyer flask of MRS liquid culture medium, cultured for 60 hours at 37 ℃, and taken out after the culture medium is turbid.
Rhodospirillum oxysporum (Ectothiorhodospirasahophikovii ACCC19809) purchased from the center for agricultural microbial culture collection in China was transferred into a flask containing a culture medium for photosynthetic bacteria, and the culture was incubated at 30 ℃ for 72 hours under light, and was taken out after the culture medium became turbid.
Taking out the above strains after the culture is finished, and then storing in a refrigerator at 2-4 ℃ for later use.
(2) Preparing a composite strain: mixing the strains obtained in the step (1) according to the following weight percentage to obtain a composite strain: 18% of rhodopseudomonas palustris, 24% of powdery pichia pastoris, 26% of bacillus amyloliquefaciens, 24% of lactobacillus plantarum and 8% of rhodospirillum sulphureum. Adding the composite strain into the culture solution according to the proportion of 1:100, and carrying out closed culture at the temperature of 30 ℃ for 7 days until the pH is less than 4.20 to obtain the zymogen liquid. Wherein, the culture solution includes: 10 parts by weight of molasses, 15 parts by weight of brown sugar and 90 parts by weight of water.
(3) Preparing a carrier: and (3) concentrating the zymophyte liquid obtained in the step (2) to 1/5 to obtain concentrated bacteria liquid (100g), adding 65g of porous starch, stirring at 100r/min and adsorbing for 1 h. And then adding 2g of beta-cyclodextrin, 4g of maltodextrin and 15g of lithium tourmaline micropowder to obtain the impregnation liquid. Then 40g of activated carbon fiber is arranged in the impregnation liquid for adsorption for 2.5h, and the supported microbial preparation is obtained by spray drying.
Example 2
The present example provides a microbial preparation, which is prepared according to the following steps:
(1) production and seed production: prepared according to the method of example 1.
(2) Preparing a composite strain: mixing the strains obtained in the step (1) according to the following weight percentage to obtain a composite strain: 12% of rhodopseudomonas palustris, 28% of powdery pichia pastoris, 22% of bacillus amyloliquefaciens, 28% of lactobacillus plantarum and 10% of rhodospirillum sulphureum. Adding the composite strain into the culture solution according to the proportion of 1:100, and carrying out closed culture at the temperature of 30 ℃ for 7 days until the pH is less than 4.20 to obtain the zymogen liquid. Wherein, the culture solution includes: 10 parts by weight of molasses, 15 parts by weight of brown sugar and 90 parts by weight of water.
(3) Preparing a carrier: and (3) concentrating the zymophyte liquid obtained in the step (2) to 1/4 to obtain concentrated bacteria liquid (100g), adding 65g of porous starch, stirring at 100r/min and adsorbing for 1 h. And then adding 2g of beta-cyclodextrin, 4g of maltodextrin and 15g of lithium tourmaline micropowder to obtain the impregnation liquid. Then 40g of activated carbon fiber is arranged in the impregnation liquid for adsorption for 2.5h, and the supported microbial preparation is obtained by spray drying.
Comparative example 1
The comparative example provides a microbial preparation, which is the concentrated bacterial liquid prepared in the example 1 of the invention.
Test example 1
In order to verify the application condition of the preparation in the culture of the penaeus vannamei boone, a test is carried out in a domestic farm in Shanghai Fengxian district in 2016 (7 months).
Test materials and methods:
1. pond: and selecting 2 mouths of the test pond, wherein the areas of the 2 mouths are respectively 17.8 mu and 18 mu, and the numbers are D1 and D2. The control pond has 2 mouths and the area is respectively 18.4 mu and 19 mu, and the numbers are D3 and D4.
2. South America white prawn larva: shrimp seeds come from Hainan island and Fujian mansion and are transferred to the ponds after local desalination and domestication reach 1.5-2 cm, and the used shrimp seeds are healthy and active and have no quality problem.
3. And (3) compound feed: the whole process uses the high-quality compound feed recommended by the Shanghai aquatic product technology promotion station.
4. Disinfection of drugs: tea seed cake, quicklime and bleaching powder are used for cleaning and disinfecting all ponds in sequence before stocking. The daily disinfection of the pond is regularly carried out by using the halogen disinfectant according to the daily condition.
5. Fertilizer: and (3) applying enough base fertilizer to each pool, applying organic fertilizer according to a constant amount, and culturing basic bait organisms. At ordinary times, the fertilizer is properly applied according to the water quality condition and the actual requirement.
6. Water for cultivation: in principle, a limited water exchange is used. The test pond is not filled with water in the middle period and does not change water, natural evaporation is properly supplemented for keeping the water level, and a small amount of water is changed and added in the later period. And the control pond can properly change water in the middle and later stages according to actual needs and the external water quality condition.
7. The microbial preparation comprises the following components: the control pond D3 used the microbial preparation provided in comparative example 1, the control pond D4 used no microbial preparation, the test pond D1 used the microbial preparation provided in example 1 of the present invention, and the test pond D2 used the microbial preparation provided in example 2 of the present invention.
The usage method of the dosage is as follows:
the microbial preparation comprises the following components: the microbial preparation used in each mu of water surface of the test ponds D1 and D2 contains 100g of concentrated bacterial liquid, and the concentrated bacterial liquid is cut into small pieces about 1cm and evenly applied.
And (3) using 100g of concentrated bacterial liquid per mu of water surface in the control pond D3, diluting by 50-100 times, and then uniformly sprinkling.
The microbial inoculum is added once a month, and the test period is 3 months (No. 7.1-No. 9.30).
8. And (3) the other: the test pond is basically the same as the control pond in the allocation, use and other facility conditions and management methods of the aerator.
(II) results and analysis
1. Stocking and harvesting conditions:
the area of the test pond D1 is 17.8 mu, 75 thousand penaeus vannamei larvae are released, 4.21 thousand penaeus vannamei larvae are released per mu, the total yield of harvest is 5696 kg, the average yield per mu is 320kg, and the average profit per mu is 2645 yuan.
The area of the test pond D1 is 18.1 mu, 76 million tails of south America white prawn seedlings are released, 4.20 million tails are released in each mu, the total yield is 5629 kg, the average yield per mu is 311kg, and the average profit per mu is 2571 yuan.
The area of the control pond D3 is 18.4 mu, 78 million tails of Penaeus vannamei Boone seedlings are released, 4.24 million tails are released in the average mu, the total yield is 4471kg, the average yield per mu is 243kg, and the average profit per mu is 1047 yuan.
The area of the control pond D4 is 19 mu, 81 million tails of south America white prawn seedlings are released, 4.26 million tails are released in the average mu, the total yield of harvest is 2685kg, the average yield per mu is 141kg, and all losses are realized.
2. The disease onset condition is as follows:
the whole process of the test ponds D1 and D2 has no disease. While the control pond D3 is partially diseased, the condition is light. The control pond D4 has a lot of diseases, is difficult to control after the diseases occur, seriously influences the harvest yield, and has to be collected in advance in the later period.
3. The water quality condition is as follows:
the water quality of the test pond is stable throughout, the composition of plankton is balanced, various conventional indexes are in a normal state for a long time, and the test pond is clean and free of dirt. The water quality of the control pond D3 in a partial area is unstable, and the whole level is unbalanced. The water quality in the control pond D4 is unstable, the bacterial facies and algal facies fluctuate violently, and the food platform inspection is always stained with a lot of dirt and dirts.
During the culture period, the four culture ponds to which the promotion stations belong are subjected to water quality index detection, and the details are shown in table 1:
TABLE 1
a: measuring by adopting a plug disc method;
b: measuring the oxidation-reduction potential at the depth of 2-3 cm;
c: the pond was collected in advance.
4. Substrate condition:
the test pond has clean substrate, less hard plate and sludge, the contrast pond D3 has uneven substrate condition, good partial condition and poor partial condition, the contrast pond D4 has poor substrate condition, thick sludge, large amount of dirt and dirt accumulated, serious blackening and stink of a feed belt, and large amount of dead shrimps accumulated.
5. Food intake:
the test pond has normal feeding of the shrimps, high feed conversion efficiency, low feed coefficient, complete digestion, absorption and utilization and obvious reduction of excrement, while the control pond D3 has abnormal feeding of part of the shrimps and the control pond D4 has relatively poor environmental conditions and usually shows abnormal feeding of the shrimps, low feed utilization rate and increased feed coefficient.
6. Growth conditions were as follows:
the test pond has good shrimp growth, good vitality, large harvest specification, healthy shrimp body and normal sensory index, the test pond is qualified through product quality sampling inspection of relevant authority departments in the market, the control pond D3 has uneven shrimp body quality, the control pond D4 is obviously influenced in growth, the harvest specification is relatively small, the individual variation is obvious, and the quality is reduced.
The embodiments described above are some, but not all embodiments of the invention. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Claims (9)

1. A microbial preparation for an ecological safety water system is characterized by being suitable for a culture water body and comprising the following components:
the composite strain consists of the following strains in percentage by weight: 15-20% of rhodopseudomonas palustris, 20-25% of powdery pichia pastoris, 25-30% of bacillus amyloliquefaciens, 5-10% of rhodococcus erythropolis, 20-25% of lactobacillus plantarum and 5-10% of rhodospirillum sulphureum; and the number of the first and second groups,
the carrier is used for loading the composite strain; the vector is obtained as follows: concentrating the zymophyte liquid of the composite strain to obtain concentrated bacteria liquid, adding porous starch, stirring and adsorbing, then adding dextrin and lithium tourmaline micropowder to obtain impregnation liquid, arranging activated carbon fibers in the impregnation liquid for adsorption for 2-3 h, and drying to obtain the activated carbon fiber-based composite strain; wherein, the stirring and adsorbing conditions are as follows: adsorbing for 0.5-1.5 h under the condition of 80-120 r/min.
2. A method for preparing a microbial preparation for an eco-safe water system according to claim 1, comprising the steps of:
s1, preparing a composite strain: mixing strains according to a ratio to obtain the composite strain, adding the composite strain into a culture solution, and carrying out closed culture at the temperature of 28-32 ℃ for 5-10 days to obtain a zymogen solution;
s2, preparing a carrier: concentrating the zymophyte liquid to obtain concentrated bacteria liquid, adding porous starch, stirring and adsorbing, then adding dextrin and lithium tourmaline micro powder to obtain impregnation liquid, arranging activated carbon fibers in the impregnation liquid, adsorbing for 2-3 h, and drying.
3. The method for producing a microbial preparation for an ecological safety water system according to claim 2, wherein in step S1, the mass ratio of the composite bacterial seed to the culture solution is 1: 80-120 parts.
4. The method for preparing a microbial preparation for an ecological safety water system according to claim 2, wherein in step S1, the culture solution comprises 8 to 12 parts by weight of molasses and 90 to 96 parts by weight of water.
5. The method for preparing a microbial preparation for an ecological safety water system according to claim 4, wherein the culture solution further comprises 10 to 15 parts by weight of brown sugar.
6. The method for preparing a microbial preparation for an eco-safe water system according to claim 2, wherein the Rhodopseudomonas palustris (Rhodopseudomonas palustris) is a strain having a collection number of ACCC10649, the Pichia farinose (Pichia farinose) is a strain having a collection number of ACCC20025, the Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) is a strain having a collection number of ACCC06476, the Rhodococcus erythropolis (Rhodococcus erythropolis) is a strain having a collection number of ACCC02667, the Lactobacillus plantarum (Lactobacillus plantarum) is a strain having a collection number of ACCC11118, and the Rhodospirillum thermosulfidoides (Ectothiophosphihoshnikvii) is a strain having a collection number of ACCC 19809.
7. The method for producing a microbial preparation for an ecological safety water system according to claim 2, wherein in step S2, the fermented bacterial liquid is concentrated to 1/4 to 1/5 in volume to obtain the concentrated bacterial liquid, and the mass ratio of the concentrated bacterial liquid to the porous starch is 1:0.5 to 0.7.
8. The method for preparing a microbial preparation for an ecological safety water system according to claim 7, wherein the dextrin comprises beta-cyclodextrin and maltodextrin in a mass ratio of 1: 1-2, and the mass ratio of the concentrated bacterial solution, the lithium tourmaline micro powder and the dextrin is 1: 0.1-0.2: 0.05-0.08.
9. The method for preparing a microbial preparation for an ecological safety water system according to claim 2, wherein the mass ratio of the concentrated bacterial liquid to the activated carbon fiber cloth is 1: 0.3-0.5.
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CN101624234A (en) * 2009-07-15 2010-01-13 南京农业大学 Microorganism water treatment agent in culturing water and preparation technique thereof
CN102703359A (en) * 2012-06-12 2012-10-03 上海亘卓生物工程有限公司 Microorganism preparation for improving cultivation water and preparation method thereof
CN102719381A (en) * 2012-06-20 2012-10-10 浙江商达环保有限公司 Composite microbial agent and preparation method and application thereof

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CN101624234A (en) * 2009-07-15 2010-01-13 南京农业大学 Microorganism water treatment agent in culturing water and preparation technique thereof
CN102703359A (en) * 2012-06-12 2012-10-03 上海亘卓生物工程有限公司 Microorganism preparation for improving cultivation water and preparation method thereof
CN102719381A (en) * 2012-06-20 2012-10-10 浙江商达环保有限公司 Composite microbial agent and preparation method and application thereof

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