CN104830733A - Microecological water purifying agent for fresh water aquiculture and preparation method thereof - Google Patents

Microecological water purifying agent for fresh water aquiculture and preparation method thereof Download PDF

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CN104830733A
CN104830733A CN201510248039.XA CN201510248039A CN104830733A CN 104830733 A CN104830733 A CN 104830733A CN 201510248039 A CN201510248039 A CN 201510248039A CN 104830733 A CN104830733 A CN 104830733A
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fermentation
cfu
preparation
aquiculture
fermentation material
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肖宇萌
沈泽竑
熊诗敏
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Hunan Hai Jia Food Science And Technology 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
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/30—Aerobic and anaerobic processes
    • 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
    • C02F3/347—Use of yeasts or fungi
    • 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
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor

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Abstract

The invention relates to a microecological purifying agent for fresh water aquiculture and a preparation method thereof, belonging to the technical field of biology in aquiculture. The method comprises the following steps: by using Bacillus subtilis, Bacillus lateraporus, microzyme and Lactobacillus acidophilus as fermentative strains, carrying out solid fermentation at proper carbon-nitrogen ratio, drying at low temperature, and adding a water purification accelerator to obtain the microecological purifying agent for fresh water aquiculture, which has the advantages of no pollution for the water body envelopment, high safety and long storage period. The microecological purifying agent for fresh water aquiculture can effectively lower the contents of organic pollutants, ammonia nitrogen, nitrite nitrogen and COD (chemical oxygen demand) in the fresh water aquiculture water body and improve the fresh water aquiculture water body envelopment. The method well overcomes the defects of poor water purification, low viable count, high production cost and the like in the product in the prior art, has the advantages of simple production technique, no discharge of three wastes and low equipment investment, and has important meanings for fresh water healthy culture and aquatic product sustainable development.

Description

A kind of freshwater aquiculture microbial water purified agents and preparation method
Technical field
The present invention relates to a kind of freshwater aquiculture microbial water purified agents and preparation method, belong to the biological technical field in aquaculture.
Background technology
Aquatic products probiotics utilizes the beneficial microorganism in the animal bodies such as fish shrimp crab or in aquaculture water or promotes the active bacteria formulation that material is formed through special processing process.Can be used for microecological regulation and control in water body, purify water, biological effect or ecologic effect can be produced, also can be used for adjustment or maintain microecological balance in animal intestinal, reach preventing disease, promote the object of aquatic health growth.Since the twentieth century middle period, along with the development of aquaculture, if the medicated feed additives such as microbiotic, olaquindox, quinoline ketone are worldwide used widely.This medicated feed additive disease preventing and treating, promote growth of animal etc. in effect fairly obvious, but consequent side effect and cause pathogenic bacteria to develop immunity to drugs, toxicity, the drawback such as " three cause " (teratogenesis, carcinogenic, mutagenesis) also reveal gradually.The problem of residual hazard, contaminate environment, threat human health has now become the focus that countries in the world cultivation worker and healthcare workers are extremely paid close attention to.Along with the widespread use on animal microecological formulation is in herding etc., the application in water industry also more and more causes the attention of people, its good effect by a large amount of experiments and production practice confirm.Probiotics has and drops into that little, income is large, without residual hazard, without resistance, the advantage such as free from environmental pollution, fully show the superiority utilizing Tiny ecosystem Prevention Technique, it must become the developing direction of 21st century culture fishery.
About the investigation and application of freshwater aquiculture probiotics, carry out a large amount of research work both at home and abroad, summed up mainly to concentrate on and utilize single microbial inoculum or composite fungus agent to improve aquaculture water water quality, or prevent some disease of aquatic animal.Domestic patent CN101717724 A, CN101735951 A, CN101878858 A, CN1498865A, CN101147529A, CN1594551A, CN101376877A, CN101629157A etc. individually disclose for water body purification of aquaculture, reparation cultivation water environment is repaired, the compound micro-ecological preparation purified water and preparation method, although these methods are except having the organic pollutant that can reduce in aquaculture water, ammonia nitrogen, nitrite, COD, BOD content etc., also have and can promote that beneficial algae is bred, improve effect of the resistance against diseases of cultivation body, but there is complex manufacturing in these patented products, product bacterial classification survival rate is low, energy consumption and cost high, the shortcomings such as result of use is unstable.
Summary of the invention
Technical problem to be solved by this invention is: for above-mentioned the deficiencies in the prior art, a kind of freshwater aquiculture microbial water purified agents and preparation method are provided, this product effectively can decompose the larger molecular organics in freshwater aquiculture water body, reduce ammonia nitrogen in water body, nitrate nitrogen content, and it is simple to have production technique, cost and energy consumption low, living bacteria count is high, strong adaptability, be easy to the feature produced in batches, solve the unstable product quality that aquatic products probiotics in the market exists, result of use is undesirable, the problem such as the low and production cost of living bacteria count survival rate is high.
In order to solve the problems of the technologies described above, the technical solution adopted in the present invention is: a kind of freshwater aquiculture microbial water purified agents and preparation method, and the method comprises following step:
(1) shake-flask seed preparation: subtilis 1 strain is inoculated a ring slant strains cell and expand in meat soup nutritional medium in liquid, at 30 DEG C-37 DEG C, under 180r/min-220r/min condition, shaking table shaking culture 16-24h, makes cell concentration in this seed culture fluid reach 10 8-10 9cfu/mL; Bacillus Lateraporus 1 strain is inoculated a ring slant strains cell in Bacillus Lateraporus liquid nutrient medium, 35 DEG C-37 DEG C, under 180r/min-220r/min condition, shaking table shaking culture 24-36h, makes cell concentration in this seed culture fluid reach 10 7-10 8cfu/mL; Lactobacterium acidophilum 1 strain is inoculated a ring slant strains cell in Lactobacterium acidophilum liquid nutrient medium, 35 DEG C-37 DEG C, quiescent culture 20-30h, make cell concentration in this seed culture fluid reach 10 7-10 8cfu/mL; Yeast saccharomyces cerevisiae 1 strain is inoculated a ring slant strains cell in yeast liquid nutrient medium, at 28 DEG C-30 DEG C, under 180r/min-220r/min condition, shaking table shaking culture 24-36h, makes cell concentration in this seed culture fluid reach 10 8-10 9cfu/mL;
(2) fermentation material preparation: get wheat bran 80%-85%, Semen Maydis powder 10%-18%, glucose 1%-2%, (NH by weight percentage 4) 2sO 41.5%-2%, KH 2pO 40.1-0.3%, MgSO 4.7H 2o 0.05-0.15%, mixes, and adjustment weight in wet base is 50%-55%, and sterilizing 60min under 121 DEG C of conditions, after being cooled to room temperature, obtains solid medium;
(3) in above-mentioned solid medium, press 15-20% respectively inoculate weight access liquid seeds, fully mix, be placed in the fermentation tray of sterilization, upper cover plastics film, 28-30 DEG C of fermentation 2-3d, then in 45-60 DEG C of dry 6-8h, pulverizing forms;
(4) get fermentation of bacillus subtilis material 3-5 part, Bacillus Lateraporus fermentation material 3-5 part, saccharomycetes to make fermentation material 2-4 part, Lactobacterium acidophilum fermentation material 6-10 part, water purification promotor 50-80 part, fully mix, get product.
The bacterial classification used in the present invention is prior art, and preserving number is respectively: subtilis (Bacillus subtilis): ACCC11062, CGMCC2548 or CGMCC1222; Bacillus Lateraporus (Bacillus lateraporus): CGMCC1755 or CGMCC9701; Lactobacterium acidophilum (Lactobacillus acidophilus): CICC6074, ACCC 11073 or CGMCC 1.2467; Saccharomyces yeast (Saccharomyces SP.): CGMCC 1147, CGMCC 0702 or CGMCC 0133.
Compared with prior art, the present invention has the following advantages:
(1) freshwater aquiculture microbial water purified agents of the present invention and preparation method can make the content of the organic pollutant in freshwater aquiculture water body, ammonia nitrogen, nitric nitrogen, COD and BOD obviously reduce lastingly;
(2) bacterial classification that the inventive method uses has included bacteria and anerobe, both can reduce the content of various objectionable impurities in aerobic water body environment, the ight soil also can degraded as feed residual in the anaerobic environments such as bed mud and aquatic animal, thus improves water quality;
(3) the inventive method is compared with existing production technique, have that production technique is simple, energy consumption is low, constant product quality, without the three wastes be easy to the features such as large-scale production;
(4) water purification promotor-peat composed of rotten mosses of using of the inventive method or polyacrylamide or zeolite powder, effectively can remove various suspended substance, COD and BOD etc. in aquaculture water, be conducive to the fast purification of microorganism to aquaculture water.
Embodiment
The invention will be further described for following examples.
Embodiment 1
The present embodiment 1 bacterial classification used is subtilis 1 strain, Bacillus Lateraporus 1 strain, Lactobacterium acidophilum 1 strain and saccharomyces yeast 1 strain:
(1) shake-flask seed preparation: subtilis 1 strain is inoculated a ring slant strains cell and expand in meat soup nutritional medium in liquid, at 32 DEG C, under 180r/min condition, shaking table shaking culture 20h, makes cell concentration in this seed culture fluid reach 2 × 10 9cfu/mL; Bacillus Lateraporus 1 strain is inoculated a ring slant strains cell in Bacillus Lateraporus liquid nutrient medium, 35 DEG C, under 200r/min condition, shaking table shaking culture 24h, makes cell concentration in this seed culture fluid reach 1.7 × 10 8cfu/mL; Lactobacterium acidophilum 1 strain is inoculated a ring slant strains cell in Lactobacterium acidophilum liquid nutrient medium, 37 DEG C, quiescent culture 30h, make cell concentration in this seed culture fluid reach 3.2 × 10 8cfu/mL; Yeast saccharomyces cerevisiae 1 strain is inoculated a ring slant strains cell in yeast liquid nutrient medium, at 28 DEG C, under 220r/min condition, shaking table shaking culture 24h, makes cell concentration in this seed culture fluid reach 4.1 × 10 9cfu/mL;
(2) fermentation material preparation: get wheat bran 85%, Semen Maydis powder 11.2%, glucose 2%, (NH by weight percentage 4) 2sO 41.5%, KH 2pO 40.15%, MgSO 4.7H 2o 0.15%, mixes, and regulate weight in wet base to be 52%, sterilizing 60min under 121 DEG C of conditions, after being cooled to room temperature, obtains solid medium;
(3) in above-mentioned solid medium, access liquid seeds by 15% inoculation weight respectively, fully mix, be placed in the fermentation tray of sterilization, upper cover plastics film, 30 DEG C of fermentation 3d, then in 50 DEG C of dry 8h, pulverizing forms;
(4) get 5 parts, fermentation of bacillus subtilis material, Bacillus Lateraporus fermentation material 4 parts, 3 parts, saccharomycetes to make fermentation material, Lactobacterium acidophilum fermentation material 8 parts, the peat composed of rotten mosses 60 parts, polyacrylamide 20 parts, fully mix, get product.
Embodiment 2:
The present embodiment 2 bacterial classification used is subtilis 1 strain, Bacillus Lateraporus 1 strain, Lactobacterium acidophilum 1 strain and saccharomyces yeast 1 strain:
(1) shake-flask seed preparation: subtilis 1 strain is inoculated a ring slant strains cell and expand in meat soup nutritional medium in liquid, at 35 DEG C, under 200r/min condition, shaking table shaking culture 18h, makes cell concentration in this seed culture fluid reach 3.3 × 10 9cfu/mL; Bacillus Lateraporus 1 strain is inoculated a ring slant strains cell in Bacillus Lateraporus liquid nutrient medium, 37 DEG C, under 180r/min condition, shaking table shaking culture 30h, makes cell concentration in this seed culture fluid reach 2.9 × 10 8cfu/mL; Lactobacterium acidophilum 1 strain is inoculated a ring slant strains cell in Lactobacterium acidophilum liquid nutrient medium, 36 DEG C, quiescent culture 25h, make cell concentration in this seed culture fluid reach 2.6 × 10 8cfu/mL; Yeast saccharomyces cerevisiae 1 strain is inoculated a ring slant strains cell in yeast liquid nutrient medium, at 30 DEG C, under 200r/min condition, shaking table shaking culture 36h, makes cell concentration in this seed culture fluid reach 3.5 × 10 8cfu/mL;
(2) fermentation material preparation: get wheat bran 82%, Semen Maydis powder 14.2%, glucose 1.5%, (NH by weight percentage 4) 2sO 42%, KH 2pO 40.2%, MgSO 4.7H 2o 0.1%, mixes, and regulate weight in wet base to be 50%, sterilizing 60min under 121 DEG C of conditions, after being cooled to room temperature, obtains solid medium;
(3) in above-mentioned solid medium, access liquid seeds by 20% inoculation weight respectively, fully mix, be placed in the fermentation tray of sterilization, upper cover plastics film, 28 DEG C of fermentation 2.5d, then in 55 DEG C of dry 6h, pulverizing forms;
(4) get 4 parts, fermentation of bacillus subtilis material, Bacillus Lateraporus fermentation material 5 parts, 4 parts, saccharomycetes to make fermentation material, Lactobacterium acidophilum fermentation material 7 parts, the peat composed of rotten mosses 45 parts, polyacrylamide 15 parts, zeolite powder 20 parts, fully mix, get product.
Embodiment 3:
The present embodiment 3 bacterial classification used is subtilis 1 strain, Bacillus Lateraporus 1 strain, Lactobacterium acidophilum 1 strain and saccharomyces yeast 1 strain:
(1) shake-flask seed preparation: subtilis 1 strain is inoculated a ring slant strains cell and expand in meat soup nutritional medium in liquid, at 37 DEG C, under 220r/min condition, shaking table shaking culture 24h, makes cell concentration in this seed culture fluid reach 6.1 × 10 9cfu/mL; Bacillus Lateraporus 1 strain is inoculated a ring slant strains cell in Bacillus Lateraporus liquid nutrient medium, 36 DEG C, under 220r/min condition, shaking table shaking culture 34h, makes cell concentration in this seed culture fluid reach 3.2 × 10 8cfu/mL; Lactobacterium acidophilum 1 strain is inoculated a ring slant strains cell in Lactobacterium acidophilum liquid nutrient medium, 35 DEG C, quiescent culture 22h, make cell concentration in this seed culture fluid reach 1.8 × 10 8cfu/mL; Yeast saccharomyces cerevisiae 1 strain is inoculated a ring slant strains cell in yeast liquid nutrient medium, at 29 DEG C, under 180r/min condition, shaking table shaking culture 30h, makes cell concentration in this seed culture fluid reach 2.7 × 10 8cfu/mL;
(2) fermentation material preparation: get wheat bran 80%, Semen Maydis powder 17%, glucose 1%, (NH by weight percentage 4) 2sO 41.8%, KH 2pO 40.15%, MgSO 4.7H 2o 0.05%, mixes, and regulate weight in wet base to be 55%, sterilizing 60min under 121 DEG C of conditions, after being cooled to room temperature, obtains solid medium;
(3) in above-mentioned solid medium, access liquid seeds by 18% inoculation weight respectively, fully mix, be placed in the fermentation tray of sterilization, upper cover plastics film, 30 DEG C of fermentation 2d, then in 60 DEG C of dry 7h, pulverizing forms;
(4) get 3 parts, fermentation of bacillus subtilis material, Bacillus Lateraporus fermentation material 5 parts, 2 parts, saccharomycetes to make fermentation material, Lactobacterium acidophilum fermentation material 10 parts, polyacrylamide 60 parts, zeolite powder 20 parts, fully mix, get product.

Claims (5)

1. freshwater aquiculture microbial water purified agents and a preparation method, is characterized in that it is made up of the raw material of following weight proportioning: fermentation of bacillus subtilis material 3-5 part, Bacillus Lateraporus fermentation material 3-5 part, saccharomycetes to make fermentation material 2-4 part, Lactobacterium acidophilum fermentation material 6-10 part, water purification promotor 50-80 part.
2. freshwater aquiculture microbial water purified agents according to claim 1 and preparation method, is characterized in that adopting following processing step:
(1) shake-flask seed preparation: subtilis 1 strain is inoculated a ring slant strains cell and expand in meat soup nutritional medium in liquid, at 30 DEG C-37 DEG C, under 180r/min-220r/min condition, shaking table shaking culture 16-24h, makes cell concentration in this seed culture fluid reach 10 8-10 9cfu/mL; Bacillus Lateraporus 1 strain is inoculated a ring slant strains cell in Bacillus Lateraporus liquid nutrient medium, 35 DEG C-37 DEG C, under 180r/min-220r/min condition, shaking table shaking culture 24-36h, makes cell concentration in this seed culture fluid reach 10 7-10 8cfu/mL; Lactobacterium acidophilum 1 strain is inoculated a ring slant strains cell in Lactobacterium acidophilum liquid nutrient medium, 35 DEG C-37 DEG C, quiescent culture 20-30h, make cell concentration in this seed culture fluid reach 10 7-10 8cfu/mL; Yeast saccharomyces cerevisiae 1 strain is inoculated a ring slant strains cell in yeast liquid nutrient medium, at 28 DEG C-30 DEG C, under 180r/min-220r/min condition, shaking table shaking culture 24-36h, makes cell concentration in this seed culture fluid reach 10 8-10 9cfu/mL;
(2) fermentation material preparation: get wheat bran 80%-85%, Semen Maydis powder 10%-18%, glucose 1%-2%, (NH by weight percentage 4) 2sO 41.5%-2%, KH 2pO 40.1-0.3%, MgSO 4.7H 2o 0.05-0.15%, mixes, and adjustment weight in wet base is 50%-55%, and sterilizing 60min under 121 DEG C of conditions, after being cooled to room temperature, obtains solid medium;
(3) in above-mentioned solid medium, press 15-20% respectively inoculate weight access liquid seeds, fully mix, be placed in the fermentation tray of sterilization, upper cover plastics film, 28-30 DEG C of fermentation 2-3d, then in 45-60 DEG C of dry 6-8h, pulverizing forms;
(4) get fermentation of bacillus subtilis material 3-5 part, Bacillus Lateraporus fermentation material 3-5 part, saccharomycetes to make fermentation material 2-4 part, Lactobacterium acidophilum fermentation material 6-10 part, water purification promotor 50-80 part, fully mix, get product.
3. freshwater aquiculture microbial water purified agents according to claim 1 and preparation method, is characterized in that living bacteria count content 1 × 10 in microbial water purified agents 9-1 × 10 10cfu/g.
4. freshwater aquiculture microbial water purified agents according to claim 1 and preparation method, is characterized in that described fermentation of bacillus subtilis material viable count content is 1 × 10 11cfu/g, Bacillus Lateraporus fermentation material viable count content is 1 × 10 11cfu/g, saccharomycetes to make fermentation material viable count content is fermentation material viable count content is 1 × 10 9cfu/g, Lactobacterium acidophilum fermentation material viable count content is fermentation material viable count content is 1 × 10 9cfu/g.
5. freshwater aquiculture microbial water purified agents according to claim 1 and preparation method, is characterized in that described water purification promotor is the peat composed of rotten mosses, polyacrylamide, zeolite powder one or more mixtures wherein.
CN201510248039.XA 2015-05-15 2015-05-15 Microecological water purifying agent for fresh water aquiculture and preparation method thereof Pending CN104830733A (en)

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106986517A (en) * 2017-03-28 2017-07-28 海南正强超越生化技术开发有限公司 A kind of cultivation substrate modifier and preparation method thereof
CN107183406A (en) * 2017-05-16 2017-09-22 山东省海洋生物研究院 A kind of microorganism live bacteria bait and its production method for shellfish hatchery
CN107285484A (en) * 2017-08-21 2017-10-24 苏州神良生物科技有限公司 A kind of freshwater aquiculture microorganism formulation
CN107445432A (en) * 2017-08-02 2017-12-08 湖北茂源水生态资源开发有限公司 A kind of riverbed sludge processing method
CN108157668A (en) * 2017-11-17 2018-06-15 辽宁景良生物科技开发有限公司 A kind of used in mariculture probiotics and its preparation method and application

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CN103304043A (en) * 2013-07-08 2013-09-18 湖南农业大学 Pool healthy culture micro-ecologic water purifier
CN104017754A (en) * 2014-05-13 2014-09-03 湖南山河美生物环保科技股份有限公司 Microbial water purifying agent and preparation method thereof
CN104560811A (en) * 2014-12-30 2015-04-29 天津市工业微生物研究所有限公司 Compound microbial agent used for treatment of aquaculture water

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103304043A (en) * 2013-07-08 2013-09-18 湖南农业大学 Pool healthy culture micro-ecologic water purifier
CN104017754A (en) * 2014-05-13 2014-09-03 湖南山河美生物环保科技股份有限公司 Microbial water purifying agent and preparation method thereof
CN104560811A (en) * 2014-12-30 2015-04-29 天津市工业微生物研究所有限公司 Compound microbial agent used for treatment of aquaculture water

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106986517A (en) * 2017-03-28 2017-07-28 海南正强超越生化技术开发有限公司 A kind of cultivation substrate modifier and preparation method thereof
CN106986517B (en) * 2017-03-28 2020-05-12 海南正强超越生化技术开发有限公司 Culture substrate modifier and preparation method thereof
CN107183406A (en) * 2017-05-16 2017-09-22 山东省海洋生物研究院 A kind of microorganism live bacteria bait and its production method for shellfish hatchery
CN107445432A (en) * 2017-08-02 2017-12-08 湖北茂源水生态资源开发有限公司 A kind of riverbed sludge processing method
CN107285484A (en) * 2017-08-21 2017-10-24 苏州神良生物科技有限公司 A kind of freshwater aquiculture microorganism formulation
CN108157668A (en) * 2017-11-17 2018-06-15 辽宁景良生物科技开发有限公司 A kind of used in mariculture probiotics and its preparation method and application

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