CN105505820A - Biological agent for relieving contamination at bottom of turbot culture pond and preparation method thereof - Google Patents

Biological agent for relieving contamination at bottom of turbot culture pond and preparation method thereof Download PDF

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Publication number
CN105505820A
CN105505820A CN201510999919.0A CN201510999919A CN105505820A CN 105505820 A CN105505820 A CN 105505820A CN 201510999919 A CN201510999919 A CN 201510999919A CN 105505820 A CN105505820 A CN 105505820A
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nitrobacteria
making method
enrichment
subtilis
weight
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江瀚
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SHANGHAI CHUANGBO ECOLOGICAL ENGINEERING Co Ltd
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SHANGHAI CHUANGBO ECOLOGICAL ENGINEERING Co Ltd
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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
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F11/00—Treatment of sludge; Devices therefor
    • C02F11/02—Biological treatment
    • 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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  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Water Supply & Treatment (AREA)
  • Environmental & Geological Engineering (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Hydrology & Water Resources (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Virology (AREA)
  • Medicinal Chemistry (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Farming Of Fish And Shellfish (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The invention discloses a biological agent for relieving contamination at the bottom of a turbot culture pond and a preparation method thereof. The preparation method is characterized by comprising the first step of culture of bacillus subtilis and nitrifying bacteria, the second step of curing of bacillus subtilis and nitrifying bacteria and the third step of preparation of the biological agent. The biological agent for relieving contamination at the bottom of the turbot culture pond is prepared through the method. Bacillus subtilis and nitrifying bacteria are adopted as the bacteria for the biological agent, and meanwhile different curing modes are adopted according to different characteristics of the two kinds of bacteria, so that effectiveness of the product is ensured; besides, the product has the advantages of accelerating the nitrogen circulation process at the bottom of the culture pond, lowering the nitrite content, inhibiting growth and propagation of harmful pathogenic bacteria at the bottom of the pond and the like.

Description

A kind ofly improve biotechnological formulation polluted bottom how precious fish culture pond and preparation method thereof
Technical field
The present invention relates to biological fodder technical field, particularly relate to and a kind ofly improve biotechnological formulation polluted bottom how precious fish culture pond and preparation method thereof.
Background technology
Many precious fishes are that Osteichthyes Pleuronectiformes Bothidae brill belongs to ocean ground fish, are commonly called as brill, claim " how precious fish " in China.
Since from the nineties, how precious fish is introduced into China, it is propagated artificially to obtain and develops rapidly, but owing to excessively to throw something and feed, the many reasons such as the shortage of some areas breeding water, the water quality of breeding water is greatly affected.
At present both at home and abroad generally single or complex microorganism water conditioner is adopted to control water quality, but because microorganism is affected by the external environment larger, anti-poor environment impact capacity is poor, once system damage is difficult to recover, therefore treatment effect is unstable, simultaneously because how precious fish is sea farming, the microbial preparation of the bottom improvement on market is mostly the fresh water seawater universal product, microbial strains is very responsive to the salinity of water body environment, causes culture propagation unsuccessful, affects product result of use.
Summary of the invention
For the problems referred to above that existing microorganism water conditioner exists, now provide a kind of and improve biotechnological formulation polluted bottom how precious fish culture pond and preparation method thereof, aim to provide the water conditioner that the how precious fish of a kind of applicable sea farming uses, to decompose the various organic pollution materials in water body and mud, purify water, reduce the nitrite content in water body, in water body, form optimum microecological balance.
Concrete technical scheme is as follows:
Improve a making method for the biotechnological formulation polluted bottom how precious fish culture pond, wherein, comprise following steps:
Step one
Step 1, is inoculated in bacillus subtilis in liquid nutrient medium and cultivates;
Step 2, is inoculated in nitrobacteria in enrichment medium and carries out enrichment culture;
Step 2
Step 1, by the subtilis culture that obtains in described step one through centrifugation, will add in zeolite powder from effluent and stir;
Step 2, makes thalline suspension liquid by the nitrobacteria enrichment culture thing obtained in above-mentioned steps one; By sodium alginate heating for dissolving in physiological saline, then mix with the thalline suspension liquid of nitrobacteria, finally under ice bath, instill calcium chloride solution, stir and obtain the spherical globule, after the described globule is placed in refrigerator crosslinking curing, then use brine.
Step 3
Step 1, stirs the cured article of subtilis and nitrobacteria cured article, the biotechnological formulation polluted bottom the how precious fish culture pond that improves.
Above-mentioned making method, preferably, in described step one, is that 8-20% is inoculated in liquid nutrient medium by subtilis according to weight percent, cultivates 24-36 hour in 35-40 DEG C.
Above-mentioned making method, preferably, in liquid nutrient medium in described step one, each component and weight percent thereof are: soybean cake powder 1.5-5%, Semen Maydis powder 2.5-7%, starch 0-4%, molasses 1-1.5%, potassium primary phosphate 0.02-0.06%, Sodium phosphate dibasic 0.2-0.6%, silicone antifoam agent 0.05-0.1%, sterilized water 80-95%.
Described nitrobacteria bacterium, preferably, in described step one, is 5-9% according to weight percent: be inoculated in enrichment medium by above-mentioned making method.
Above-mentioned making method, preferably, in enrichment medium in described step one, each component and weight percent thereof are: ammonium sulfate 0.3-0.7%, sodium bicarbonate 0.6-1.4%, sodium-chlor 0.1-0.7%, magnesium sulfate heptahydrate 0.01-0.07%, iron vitriol 0.01-0.03%, Calcium dichloride dihydrate 0.06-0.1%, glucose 0.05-0.1%, sterilized water 96-98%.
Above-mentioned making method, preferably, in described step one, be inoculated in by nitrobacteria in enrichment medium, control pH is 7.5-8.5, and control temperature is 28-30 DEG C, Vs30-40%, (Do) > 2mgL-1, carries out cultured continuously enrichment 14-21 days.
Above-mentioned making method, preferably, in described step 2, subtilis culture is separated through disk centrifugal separator, centrifuge speed is 7000 rev/min, centrifugal thing higher for the gemma purity obtained is added in 60 order zeolite powders according to the part by weight of 1:4-5, stirs, control moisture weight mark at 15-20%.
Above-mentioned making method, preferably, in described step 2, by the nitrobacteria of enrichment with 0.9% physiological saline centrifuge washing 2-3 time, centrifuge speed is 4000 rev/min, each 15 minutes, use normal saline dilution 2-3 doubly again, be uniformly mixed, make thalline suspension liquid; By sodium alginate according to the part by weight heating for dissolving of 5-10% in the physiological saline of 0.9%, then mix according to the part by weight of 1:1-2 with the thalline suspension liquid of nitrobacteria, the calcium chloride solution of 0.1mg/L is instilled under last ice bath, stir, obtain the spherical globule of diameter 2mm, the described globule to be positioned in 4 DEG C of refrigerators crosslinking curing after 24 hours, then to use brine 2-3 time.
Above-mentioned making method, preferably, in described step 3, is 6:1 mixing by the cured article of subtilis and the cured article of nitrobacteria according to weight ratio.
According to above-mentioned any one the how precious fish culture pond of improvement that makes of making method bottom the biotechnological formulation that pollutes.
It should be noted that when not having specified otherwise, the concept such as ratio, content described herein, all refers to part by weight or weight content.
The beneficial effect of technique scheme:
The bacterial classification that the present invention adopts is Bacillus subtillis, nitrobacteria, and the characteristic different according to two bacterial classifications have employed different curing modes simultaneously, has ensured the validity of product.Biotechnological formulation advantage provided by the present invention is as follows:
1, accelerate the nitrogen cycle process bottom cultivating pool, reduce nitrite content.
2, adsorbable and transform bottom organic dirt and the content of heavy metal ion.
3, the NO of exploded bottom 2-, H 2the harmful chemicals such as S, suspension organic matter, residual bait and movement effectively in degraded water body, reduce the formation that bottom is polluted.
4, beneficial microorganism dominant population can be formed after using, suppress pond bottom to be harmful to the growth and breeding of pathogenic bacteria.
5, utilize composite curing technology, Multiple components is fixed on special carrier, and microorganism and metabolite mass-energy thereof are stablized in one relatively long-time play a role, to saline environment in resistant to sea water.
Embodiment
Below in conjunction with the embodiment of the present invention, the technical scheme in the embodiment of the present invention is clearly and completely described.
Embodiment one one kinds improves the biotechnological formulation polluted bottom how precious fish culture pond
Improve the biotechnological formulation polluted bottom how precious fish culture pond, wherein, obtained by following steps:
The cultivation of step one, subtilis and nitrobacteria
Step 1, be that 8%:92% inoculates by bacillus subtilis and liquid nutrient medium according to weight percent, cultivate 24 hours in 35 DEG C in liquid nutrient medium, wherein, in liquid nutrient medium, each component and weight percent thereof are: soya-bean cake class 1.5%, Semen Maydis powder 2.5%, starch 2%, molasses 1%, potassium primary phosphate 0.02%, Sodium phosphate dibasic 0.2%, silicone antifoam agent 0.05%, sterilized water 92.73%;
Step 2, is that 5%:95% inoculates by nitrobacteria and enrichment medium according to weight percent, in enrichment medium, carries out enrichment culture, and control pH is 7.5, control temperature 28 DEG C, Vs:30%, (Do): > 2mgL -1incubation time is 14 days, wherein, in enrichment medium, each component and total amount per-cent thereof are: ammonium sulfate 0.49%, sodium bicarbonate 0.6%, sodium-chlor 0.1%, magnesium sulfate heptahydrate 0.01%, iron vitriol 0.01%, Calcium dichloride dihydrate 0.1%, glucose 0.1%, sterilized water 98%;
The solidification of step 2, subtilis and nitrobacteria
Step 1, the subtilis culture obtained in step one is separated through disk centrifugal separator, centrifuge speed is 7000 rev/min, obtain the centrifugal thing that gemma purity is higher, by from the part by weight of effluent according to 1:4, add 60 object zeolite powders to stir, control moisture 15%;
Step 2, by the nitrobacteria of enrichment culture with 0.9% physiological saline centrifuge washing 2 times, centrifuge speed is 4000 rev/min, each 15 minutes, then uses normal saline dilution 2 times, stirs, mixes, make thalline suspension liquid;
By sodium alginate according to 5% ratio heating for dissolving in the physiological saline of 0.9%, then mix according to the part by weight of 1:1 with nitrobacteria thalline suspension liquid, finally instill the calcium chloride solution of 0.1mg/L, ice bath, drip while stir; Make it form the 2mm diameter spherical globule, the bead of formation to be positioned in 4 DEG C of refrigerators crosslinking curing after 24 hours, with brine 2 times;
The preparation of step 3, biological reagent
Step 1, mixes the cured article of subtilis with the ratio of nitrobacteria cured article according to 6:1, the biotechnological formulation polluted bottom the how precious fish culture pond that improves.
Embodiment 21 kinds improves the biotechnological formulation polluted bottom how precious fish culture pond
Improve the biotechnological formulation polluted bottom how precious fish culture pond, wherein, obtained by following steps:
The cultivation of step one, subtilis and nitrobacteria
Step 1, be that 20%:80% inoculates by bacillus subtilis and liquid nutrient medium according to weight percent, cultivate 36 hours in 40 DEG C in liquid nutrient medium, wherein, in liquid nutrient medium, each component and weight percent thereof are: soya-bean cake class 5%, Semen Maydis powder 7%, starch 4%, molasses 1.5%, potassium primary phosphate 0.06%, Sodium phosphate dibasic 0.6%, silicone antifoam agent 0.1%, sterilized water 81.74%;
Step 2, be that 9%:91% inoculates by nitrobacteria and enrichment medium according to weight percent, enrichment culture is carried out in enrichment medium, and control pH is 8.5, control temperature 30 DEG C, Vs:40%, (Do): > 2mgL-1, incubation time is 21 days, wherein, in enrichment medium and total amount per-cent be: ammonium sulfate 0.7%, sodium bicarbonate 1.4%, sodium-chlor 0.7%, magnesium sulfate heptahydrate 0.07%, iron vitriol 0.03%, Calcium dichloride dihydrate 0.1%, glucose 0.1%, sterilized water 96.9%;
The solidification of step 2, subtilis and nitrobacteria
Step 1, is separated the subtilis culture obtained in step one through disk centrifugal separator, and centrifuge speed is 7000 rev/min, obtain the centrifugal thing that gemma purity is higher, by from the part by weight of effluent according to 1:5, add 60 object zeolite powders and stir, control moisture 20%;
Step 2, by the nitrobacteria of enrichment culture that obtains in step one with 0.9% physiological saline centrifuge washing 3 times, centrifuge speed is 4000 rev/min, each 15 minutes, then uses normal saline dilution 3 times, stirs, mixes, make thalline suspension liquid;
By sodium alginate according to 10% ratio heating for dissolving in the physiological saline of 0.9%, then mix according to the part by weight of 1:2 with nitrobacteria thalline suspension liquid, finally instill the calcium chloride solution of 0.1mg/L, ice bath, drip while stir; Make it form the 2mm diameter spherical globule, the bead of formation to be positioned in 4 DEG C of refrigerators crosslinking curing after 24 hours, with brine 2 times;
The preparation of step 3, biological reagent
Step 1, mixes the cured article of subtilis with the part by weight of nitrobacteria cured article according to 6:1, the biotechnological formulation polluted bottom the how precious fish culture pond that improves.
Embodiment 31 kinds improves the biotechnological formulation polluted bottom how precious fish culture pond
Improve the biotechnological formulation polluted bottom how precious fish culture pond, wherein, obtained by following steps:
The cultivation of step one, subtilis and nitrobacteria
Step 1, be that 15%:85% inoculates by bacillus subtilis and liquid nutrient medium according to weight percent, cultivate 30 hours in 37 DEG C in liquid nutrient medium, wherein, in liquid nutrient medium, each component and weight percent thereof are: soya-bean cake class 4%, Semen Maydis powder 6%, starch 3%, molasses 1.2%, potassium primary phosphate 0.04%, Sodium phosphate dibasic 0.4%, silicone antifoam agent 0.06%, sterilized water 85.3%;
Step 2, be that 7%:93% inoculates by nitrobacteria and enrichment medium according to weight percent, enrichment culture is carried out in enrichment medium, and control pH is 8.0, control temperature 30 DEG C, Vs:35%, (Do): > 2mgL-1, incubation time is 21 days, wherein, in enrichment medium, each component and total amount per-cent thereof are: ammonium sulfate 0.5%, sodium bicarbonate 1.0%, sodium-chlor 0.5%, magnesium sulfate heptahydrate 0.05%, iron vitriol 0.02%, Calcium dichloride dihydrate 0.07%, glucose 0.08%, sterilized water 97.78%;
The solidification of step 2, cultured product
Step 1, is separated the subtilis culture obtained in step one through disk centrifugal separator, and centrifuge speed is 7000r/min, obtain the centrifugal thing that gemma purity is higher, by from the ratio of effluent according to 1:4.5, add 60 object zeolite powders and stir, control moisture 17%;
Step 2, by the nitrobacteria of enrichment culture with 0.9% physiological saline centrifuge washing 3 times, centrifuge speed is 4000 rev/min, each 15 minutes, then uses normal saline dilution 3 times, stirs, mixes, make thalline suspension liquid;
By sodium alginate according to 6% ratio heating for dissolving in the physiological saline of 0.9%, then mix according to the ratio of 1:1.5 with nitrobacteria thalline suspension liquid, finally instill the calcium chloride solution of 0.1mg/L, ice bath, drip while stir; Make it form the 2mm diameter spherical globule, the bead of formation to be positioned in 4 DEG C of refrigerators crosslinking curing after 24 hours, with brine 3 times;
The preparation of step 3, biological reagent
Step 1, mixes the cured article of subtilis with the ratio of nitrobacteria cured article according to 6:1, the biotechnological formulation polluted bottom the how precious fish culture pond that improves.
These are only preferred embodiment of the present invention; not thereby embodiments of the present invention and protection domain is limited; to those skilled in the art; should recognize and all should be included in the scheme that equivalent replacement done by all utilizations description of the present invention and apparent change obtain in protection scope of the present invention.

Claims (10)

1. improve a making method for the biotechnological formulation polluted bottom how precious fish culture pond, it is characterized in that, comprise following steps:
Step one
Step 1, is inoculated in bacillus subtilis in liquid nutrient medium and cultivates;
Step 2, is inoculated in nitrobacteria in enrichment medium and carries out enrichment culture;
Step 2
Step 1, by the subtilis culture that obtains in described step one through centrifugation, will add in zeolite powder from effluent and stir;
Step 2, makes thalline suspension liquid by the nitrobacteria enrichment culture thing obtained in above-mentioned steps one; By sodium alginate heating for dissolving in physiological saline, then mix with the thalline suspension liquid of nitrobacteria, finally under ice bath, instill calcium chloride solution, stir and obtain the spherical globule, after the described globule is placed in refrigerator crosslinking curing, then use brine.
Step 3
Step 1, stirs the cured article of subtilis and nitrobacteria cured article, the biotechnological formulation polluted bottom the how precious fish culture pond that improves.
2. making method according to claim 1, is characterized in that, in described step one, is that 8-20% is inoculated in liquid nutrient medium by subtilis according to weight percent, cultivates 24-36 hour in 35-40 DEG C.
3. making method according to claim 1 and 2, it is characterized in that, in described liquid nutrient medium, each component and weight percent thereof are: soybean cake powder 1.5-5%, Semen Maydis powder 2.5-7%, starch 0-4%, molasses 1-1.5%, potassium primary phosphate 0.02-0.06%, Sodium phosphate dibasic 0.2-0.6%, silicone antifoam agent 0.05-0.1%, sterilized water 80-95%.
4. making method according to claim 1, is characterized in that, in described step one, is that 5-9% is inoculated in enrichment medium by described nitrobacteria bacterium according to weight percent.
5. the making method according to claim 1 or 4, it is characterized in that, in described enrichment medium, each component and weight percent thereof are: ammonium sulfate 0.3-0.7%, sodium bicarbonate 0.6-1.4%, sodium-chlor 0.1-0.7%, magnesium sulfate heptahydrate 0.01-0.07%, iron vitriol 0.01-0.03%, Calcium dichloride dihydrate 0.06-0.1%, glucose 0.05-0.1%, sterilized water 96-98%.
6. the making method according to claim 1 or 4, is characterized in that, in described step one, be inoculated in by nitrobacteria in enrichment medium, control pH is 7.5-8.5, and control temperature is 28-30 DEG C, Vs30-40%, (Do) > 2mgL -1, carry out cultured continuously enrichment 14-21 days.
7. making method according to claim 1, it is characterized in that, in described step 2, subtilis culture is separated through disk centrifugal separator, centrifuge speed is 7000 rev/min, by centrifugal thing higher for the gemma purity that obtains, adds in 60 order zeolite powders according to the part by weight of 1:4-5, stir, control moisture weight mark at 15-20%.
8. making method according to claim 1, it is characterized in that, in described step 2, by the nitrobacteria of enrichment with 0.9% physiological saline centrifuge washing 2-3 time, centrifuge speed is 4000 rev/min, each 15 minutes, then uses normal saline dilution 2-3 doubly, be uniformly mixed, make thalline suspension liquid; By sodium alginate according to the part by weight heating for dissolving of 5-10% in the physiological saline of 0.9%, then mix according to the part by weight of 1:1-2 with the thalline suspension liquid of nitrobacteria, the calcium chloride solution of 0.1mg/L is instilled under last ice bath, stir, obtain the spherical globule of diameter 2mm, the described globule to be positioned in 4 DEG C of refrigerators crosslinking curing after 24 hours, then to use brine 2-3 time.
9. making method according to claim 1, is characterized in that, in described step 3, is 6:1 mixing by the cured article of subtilis and the cured article of nitrobacteria according to weight ratio.
10. according in claim 1-9 described in any one the how precious fish culture pond of improvement that makes of making method bottom the biotechnological formulation that pollutes.
CN201510999919.0A 2015-12-28 2015-12-28 Biological agent for relieving contamination at bottom of turbot culture pond and preparation method thereof Pending CN105505820A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106721523A (en) * 2016-11-23 2017-05-31 青岛水态宝生物科技有限公司 A kind of probiotics for being applied to turbot cultivation
CN108328749A (en) * 2018-04-24 2018-07-27 福州科力恩生物科技有限公司 A kind of device and its processing method using immobilization biological filler treated sewage

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CN1990853A (en) * 2005-12-28 2007-07-04 上海创博生态工程有限公司 Microorganism nitrite degradation agent and preparation method thereof

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CN1990853A (en) * 2005-12-28 2007-07-04 上海创博生态工程有限公司 Microorganism nitrite degradation agent and preparation method thereof

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106721523A (en) * 2016-11-23 2017-05-31 青岛水态宝生物科技有限公司 A kind of probiotics for being applied to turbot cultivation
CN106721523B (en) * 2016-11-23 2020-11-06 青岛家宝生物科技有限公司 Micro-ecological preparation applied to turbot culture
CN108328749A (en) * 2018-04-24 2018-07-27 福州科力恩生物科技有限公司 A kind of device and its processing method using immobilization biological filler treated sewage

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Application publication date: 20160420