CN108726692B - Aquaculture antidote - Google Patents
Aquaculture antidote Download PDFInfo
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- CN108726692B CN108726692B CN201810623546.0A CN201810623546A CN108726692B CN 108726692 B CN108726692 B CN 108726692B CN 201810623546 A CN201810623546 A CN 201810623546A CN 108726692 B CN108726692 B CN 108726692B
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- bacillus coagulans
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- 238000009360 aquaculture Methods 0.000 title claims abstract description 35
- 244000144974 aquaculture Species 0.000 title claims abstract description 35
- 239000000729 antidote Substances 0.000 title claims abstract description 30
- 239000007788 liquid Substances 0.000 claims abstract description 65
- 238000000855 fermentation Methods 0.000 claims abstract description 52
- 230000004151 fermentation Effects 0.000 claims abstract description 52
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 48
- 241000194031 Enterococcus faecium Species 0.000 claims abstract description 40
- 241000193749 Bacillus coagulans Species 0.000 claims abstract description 33
- 229940054340 bacillus coagulans Drugs 0.000 claims abstract description 33
- 239000002131 composite material Substances 0.000 claims abstract description 25
- 238000012258 culturing Methods 0.000 claims abstract description 20
- 239000006228 supernatant Substances 0.000 claims abstract description 16
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 11
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 11
- 108090000790 Enzymes Proteins 0.000 claims abstract description 10
- 102000004190 Enzymes Human genes 0.000 claims abstract description 10
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 10
- 239000003910 polypeptide antibiotic agent Substances 0.000 claims abstract description 10
- 239000002994 raw material Substances 0.000 claims abstract description 8
- 239000001963 growth medium Substances 0.000 claims description 25
- 238000009630 liquid culture Methods 0.000 claims description 18
- 230000001580 bacterial effect Effects 0.000 claims description 17
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 12
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 12
- 238000011081 inoculation Methods 0.000 claims description 12
- 238000002360 preparation method Methods 0.000 claims description 11
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 9
- 239000001888 Peptone Substances 0.000 claims description 8
- 108010080698 Peptones Proteins 0.000 claims description 8
- 238000002156 mixing Methods 0.000 claims description 8
- 235000019319 peptone Nutrition 0.000 claims description 8
- 239000007787 solid Substances 0.000 claims description 8
- 238000003756 stirring Methods 0.000 claims description 7
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 6
- 229940041514 candida albicans extract Drugs 0.000 claims description 6
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 6
- 239000008103 glucose Substances 0.000 claims description 6
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 6
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 6
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 6
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 6
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 6
- 239000012138 yeast extract Substances 0.000 claims description 6
- 244000061458 Solanum melongena Species 0.000 claims description 5
- 235000002597 Solanum melongena Nutrition 0.000 claims description 5
- 238000004140 cleaning Methods 0.000 claims description 5
- 238000005273 aeration Methods 0.000 claims description 4
- 238000010899 nucleation Methods 0.000 claims description 4
- 238000005406 washing Methods 0.000 claims description 4
- 229920001817 Agar Polymers 0.000 claims description 3
- 235000019764 Soybean Meal Nutrition 0.000 claims description 3
- 240000008042 Zea mays Species 0.000 claims description 3
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 3
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 3
- 239000008272 agar Substances 0.000 claims description 3
- 239000003674 animal food additive Substances 0.000 claims description 3
- 235000015278 beef Nutrition 0.000 claims description 3
- 235000005822 corn Nutrition 0.000 claims description 3
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 3
- 235000019797 dipotassium phosphate Nutrition 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- 239000000284 extract Substances 0.000 claims description 3
- 229940099596 manganese sulfate Drugs 0.000 claims description 3
- 239000011702 manganese sulphate Substances 0.000 claims description 3
- 235000007079 manganese sulphate Nutrition 0.000 claims description 3
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- 239000004455 soybean meal Substances 0.000 claims description 3
- 235000012054 meals Nutrition 0.000 claims description 2
- 230000001105 regulatory effect Effects 0.000 claims description 2
- 230000000593 degrading effect Effects 0.000 abstract description 7
- 230000000694 effects Effects 0.000 abstract description 7
- 244000000010 microbial pathogen Species 0.000 abstract description 7
- 230000002401 inhibitory effect Effects 0.000 abstract description 5
- 230000000087 stabilizing effect Effects 0.000 abstract description 5
- 239000000126 substance Substances 0.000 abstract description 5
- 241000894006 Bacteria Species 0.000 description 12
- 244000005700 microbiome Species 0.000 description 8
- 239000000243 solution Substances 0.000 description 7
- 230000001954 sterilising effect Effects 0.000 description 7
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 6
- 239000001301 oxygen Substances 0.000 description 6
- 229910052760 oxygen Inorganic materials 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 5
- 238000001816 cooling Methods 0.000 description 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 4
- 230000000844 anti-bacterial effect Effects 0.000 description 3
- 229940075522 antidotes Drugs 0.000 description 3
- 238000011049 filling Methods 0.000 description 3
- 231100000956 nontoxicity Toxicity 0.000 description 3
- 230000002829 reductive effect Effects 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- JVTAAEKCZFNVCJ-REOHCLBHSA-N L-lactic acid Chemical compound C[C@H](O)C(O)=O JVTAAEKCZFNVCJ-REOHCLBHSA-N 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 239000000498 cooling water Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000012851 eutrophication Methods 0.000 description 2
- 210000001035 gastrointestinal tract Anatomy 0.000 description 2
- 230000002147 killing effect Effects 0.000 description 2
- 239000004310 lactic acid Substances 0.000 description 2
- 235000014655 lactic acid Nutrition 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 238000005067 remediation Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
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- 241000186361 Actinobacteria <class> Species 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000238557 Decapoda Species 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 241000108664 Nitrobacteria Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 230000003631 expected effect Effects 0.000 description 1
- 238000005189 flocculation Methods 0.000 description 1
- 230000016615 flocculation Effects 0.000 description 1
- 235000013312 flour Nutrition 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 230000000243 photosynthetic effect Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 231100000572 poisoning Toxicity 0.000 description 1
- 230000000607 poisoning effect Effects 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
Classifications
-
- 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
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/44—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a nitrogen atom attached to the same carbon skeleton by a single or double bond, this nitrogen atom not being a member of a derivative or of a thio analogue of a carboxylic group, e.g. amino-carboxylic acids
- A01N37/46—N-acyl derivatives
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N59/00—Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/50—Treatment of water, waste water, or sewage by addition or application of a germicide or by oligodynamic treatment
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/52—Treatment of water, waste water, or sewage by flocculation or precipitation of suspended impurities
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/20—Nature of the water, waste water, sewage or sludge to be treated from animal husbandry
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Hydrology & Water Resources (AREA)
- Environmental & Geological Engineering (AREA)
- Water Supply & Treatment (AREA)
- Organic Chemistry (AREA)
- Plant Pathology (AREA)
- Wood Science & Technology (AREA)
- Pest Control & Pesticides (AREA)
- Agronomy & Crop Science (AREA)
- Health & Medical Sciences (AREA)
- Dentistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Inorganic Chemistry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
- Farming Of Fish And Shellfish (AREA)
Abstract
The invention discloses an aquaculture antidote which comprises the following raw materials in parts by weight: 5-15% of active carbon, 5-15% of biological enzyme, 20-30% of diatomite, 10-20% of antibacterial peptide and the balance of composite fermentation liquor; wherein the composite fermentation liquid is supernatant liquid obtained by mixed fermentation of bacillus coagulans and enterococcus faecium. The aquaculture antidote disclosed by the invention utilizes the supernatant obtained by carrying out submerged fermentation by co-culturing bacillus coagulans and enterococcus faecium, and is matched with active carbon, biological enzyme, diatomite and antibacterial peptide, so that the antidote has the effects of degrading organic matters in aquaculture water, decomposing and converting harmful substances, stabilizing the pH value of the water and inhibiting pathogenic microorganisms.
Description
Technical Field
The invention relates to the technical field of aquaculture preparations, in particular to an aquaculture antidote.
Background
With the mass appearance of the large-scale aquaculture industry, the aquaculture density is continuously increased, the excrement in the aquaculture water body and the residue of bait are continuously increased, pathogenic microorganisms are propagated in a large quantity, the eutrophication degree of the aquaculture water body is increased, and the ecological environment is seriously damaged, so that various diseases and emergency states frequently occur to aquatic animals in a large scale.
As is well known, when water quality is in a problem in aquaculture, an antidote is always used for adjusting the water quality, so that pathological changes of aquatic animals caused by poisoning are avoided; the microbial remediation technology is mainly applied to in-situ remediation of aquaculture environment in aquaculture and mainly solves the problems of organic pollution of bottom mud and eutrophication of water. Researches prove that the microorganisms applied to the aquaculture water body have the effects of degrading organic matters, decomposing and converting harmful substances, stabilizing the pH value of the water body and inhibiting pathogenic microorganisms.
At present, microecologics in the market have more commodities, main beneficial bacteria of the microecologics comprise photosynthetic bacteria, yeast bacteria, bacillus bacteria, actinomycetes, lactic acid bacteria, nitrobacteria, denitrifying bacteria, effective microbial flora and the like, and the effects of the probiotics in aquaculture are mainly reflected in three aspects of nutrition, immunity and ecological environment improvement. Although the microecologics have certain effect in aquaculture, the bacterial strains are not inherent bacteria in the water environment, can not survive in the water environment and the intestinal tracts of cultured animals, and can not successfully breed into dominant flora for regulation, and researches suggest that the added microorganisms are easy to gradually die over time. Meanwhile, even if indigenous bacteria are separated from the culture water environment or the intestinal tracts of aquatic animals as beneficial bacteria, the expected effect cannot be achieved due to the lack of a theoretical guide of sufficient maturity. In addition, the variety of usable substrates of a single microorganism is limited, and organic matters in the water body cannot be completely degraded, so that different microorganisms need to be compatible and act synergistically. By adjusting the microecological balance in the animal body, the occurrence of viral diseases is reduced, the abuse of antibiotics is avoided, and the breeding safety is improved.
Therefore, the invention provides an aquaculture antidote, which is prepared by carrying out deep fermentation after compatibility of different microorganisms, wherein the obtained antidote has high viable bacteria content, and has the effects of degrading organic matters in aquaculture water, decomposing and converting harmful substances, stabilizing the pH value of the water and inhibiting pathogenic microorganisms.
Disclosure of Invention
In view of the above, the invention provides an aquaculture antidote, which is prepared by carrying out deep fermentation after compatibility of different microorganisms, has high viable count, and can be used for degrading organic matters in aquaculture water, decomposing and converting harmful substances, stabilizing the pH value of the water, inhibiting pathogenic microorganisms, purifying the water, effectively increasing dissolved oxygen in the water and improving water quality after being compatible with other components.
In order to achieve the purpose, the invention adopts the following technical scheme:
an aquaculture antidote is characterized by comprising the following raw materials in parts by weight: 5-15% of active carbon, 5-15% of biological enzyme, 20-30% of diatomite, 10-20% of antibacterial peptide and the balance of composite fermentation liquor; wherein the composite fermentation liquid is the supernatant liquid obtained by mixing and fermenting bacillus coagulans and enterococcus faecium.
The invention utilizes the compatibility of different microorganisms, carries out submerged fermentation, and is compatible with active carbon, biological enzyme, diatomite, antibacterial peptide and other components, and the generated antidote has the characteristics of no pollution, no toxicity, good stability, high safety performance, high antibacterial efficiency and the like. In addition, the antidote also has the performance of increasing the dissolved oxygen in water, improving water quality, absorbing and degrading organic matters in water and effectively sterilizing.
Further, the preparation method of the composite fermentation liquor in the aquaculture antidote comprises the following steps:
(1) culturing bacillus coagulans to generate bacillus coagulans seed liquid;
(2) culturing enterococcus faecium to generate enterococcus faecium seed liquid;
(3) inoculating the bacillus coagulans seed liquid and the enterococcus faecium seed liquid into the same fermentation tank for mixed fermentation;
(4) and (4) preparing the composite fermentation liquor.
The bacillus coagulans and the enterococcus faecium are co-cultured, and the generated composite fermentation liquid is used for preparing the aquaculture antidote. Wherein the Bacillus coagulans decomposes saccharides to produce L-lactic acid; enterococcus faecium can also generate lactic acid, and the enterococcus faecium is a kind of lactobacillus with strong stress resistance and has good high temperature resistance, acid-base resistance, high salt resistance and other properties; meanwhile, the adhesive capacity is good, the tolerance is good, the effect of replacing organic acid by acidic supernatant generated by mixing and fermenting the two strains is clear, and the functions of improving water quality, killing bacteria in water and inhibiting the growth of harmful bacteria are achieved.
Further, the preparation of the bacillus coagulans seed solution comprises the following steps:
transferring the preserved bacillus coagulans to a freshly prepared solid culture medium, culturing for 24 hours at 30-37 ℃, transferring the slant strain to an eggplant bottle slant, culturing for 24 hours at 30-37 ℃, cleaning the solid culture medium, and collecting lawn washing liquid;
b, inoculating the lawn washing liquid obtained in the step a into a liquid culture medium in a seed tank, carrying out deep aeration stirring culture for 16-24 hours at the temperature of 30-38 ℃ and at the pressure of 0.04-0.06MPa and at the speed of 120-;
further, the components of the solid medium in the step a comprise: 8-10g of peptone, 5-7g of beef extract, 5-7g of sodium chloride and 18-20g of agar, fixing the volume to 1000ml by using distilled water, and adjusting the pH value to 7.2-7.4.
The components of the liquid culture medium in the step b comprise: 8-20g of peptone, 5-15g of yeast extract, 0.2-2g of dipotassium phosphate, 0.2-2g of monopotassium phosphate, 0.1-2g of magnesium sulfate, 8-20g of glucose, 1-10g of calcium carbonate and 1000ml of water.
Further, the preparation method of the enterococcus faecium seed liquid comprises the following steps:
inoculating enterococcus faecium to a liquid culture medium in a triangular flask, culturing for 14-24 hours at 30-38 ℃ under anaerobic conditions, transferring the grown enterococcus faecium liquid into another triangular flask liquid culture medium with the inoculation amount of 5-10%, and statically culturing for 14-24 hours at 30-38 ℃;
b, transferring the other triangular flask bacterial liquid into a liquid culture medium of a seeding tank according to the inoculation amount of 5-10%, and statically culturing for 14-24 hours at 30-38 ℃ and 0.04-0.06MPa to obtain the enterococcus faecium seed liquid; wherein the pH value of the enterococcus faecium seed liquid is 4.5-5.0, and the bacterial count reaches 5-15 hundred million cfu/ml.
Further, the components of the liquid medium include: 5-15g of peptone, 0.5-5g of glucose, 4-6g of corn meal, 4-10g of soybean meal, 4-6g of yeast extract, 0.5-1.5g of monopotassium phosphate, 0.3-1g of magnesium sulfate, 5-20mg of manganese sulfate and 1000ml of water.
Further, the specific steps of step (3) are as follows:
transferring the enterococcus faecium seed liquid into a fermentation tank by an inoculation amount of 5-10%, and statically culturing at 30-38 ℃ and 0.04-0.06MPa until the enterococcus faecium reaches the logarithmic phase;
b, adjusting the pH value in the fermentation tank to 7.2-7.4; adding 1-10g/L calcium carbonate and 0.2-2g/L dipotassium hydrogen phosphate;
c inoculating the bacillus coagulans seed solution into a fermentation tank with the inoculation amount of 5-10%, keeping the temperature at 30-38 ℃, the pressure at 0.04-0.06MPa and the rotation/minute at 120-.
Further, the specific steps of step (4) are as follows: centrifuging when the spore rate of the bacillus coagulans in the fermentation tank liquid reaches 90% -95%, and collecting concentrated bacterial liquid and fermentation supernatant; drying the concentrated bacterial liquid and then using the dried concentrated bacterial liquid to prepare a feed additive; and regulating the pH value of the fermentation supernatant to 3.5-4.5 by using hydrochloric acid to obtain the composite fermentation liquid.
According to the technical scheme, compared with the prior art, the aquaculture antidote provided by the invention has the following technical advantages:
(1) the antidote disclosed by the invention has the characteristics of no pollution and no toxicity, good stability, high safety performance, high antibacterial efficiency and the like.
(2) The antidote disclosed by the invention has the characteristics of water purification, water pH stabilization, strong flocculation capability, high efficiency and the like.
(3) In addition, the antidote also has the performance of increasing the dissolved oxygen in water, improving water quality, absorbing and degrading organic matters in water and effectively sterilizing.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. 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.
Example 1
Preparation of Bacillus coagulans seed solution
The Bacillus coagulans (Bacillus coagulans) in the embodiment is purchased from Guangdong province microorganism culture collection center, and the strain number is GDMCC 1.645.
a. And (3) strain culture: the preserved bacillus coagulans is transferred to a test tube slant placed in a freshly prepared solid culture medium and cultured for 24 hours at 37 ℃, and 2 eggplant bottle slants of 250ml are transferred to the slant of the slant strain and cultured for 24 hours at 37 ℃.
b. Seed tank culture: adding a liquid culture medium into a seed tank, wherein the liquid loading amount is 40%, adjusting the pH value to 7.6, stirring, sterilizing at 121 ℃ for 30 minutes, cooling by using cooling water, introducing sterile air to control the tank pressure to be 0.05MPa, when the temperature is reduced to 36 ℃, cleaning the bacteria on 2 eggplant bottles which are well grown by using 200ml of sterile physiological saline, collecting bacterial lawn cleaning solution, inoculating the bacterial lawn cleaning solution into the liquid culture medium in the seed tank, keeping the temperature at 37 ℃ and the tank pressure at 0.05MPa, and stirring at the speed of 120 rpm, carrying out deep-layer aeration stirring culture for 24 hours, and obtaining the bacillus coagulans seed liquid when the number of bacillus coagulans in the culture liquid in the seed tank is not less than 20-30 hundred million cfu/ml.
The preparation method of the solid culture medium comprises the steps of preparing peptone 10g, beef extract 5g, sodium chloride 5g, distilled water to a constant volume of 1000ml, adjusting pH to 7.2, adding agar 18g, heating to melt, filling into test tubes of 18mm x 180mm, filling eggplant bottles of 250ml in each tube of 8ml, filling 85ml in each tube, sterilizing at 121 ℃ for 30 minutes, cooling to 50 ℃ and placing on an inclined plane for later use.
The formula of the liquid culture medium comprises 20g of protein, 10g of yeast extract, 1g of dipotassium phosphate, 1g of monopotassium phosphate, 0.5g of magnesium sulfate, 20g of glucose, 8g of calcium carbonate and 1000ml of water.
Example 2
Preparation of enterococcus faecium seed liquid
Enterococcus faecium (enterococcus faecium) in this example was purchased from the Guangdong province collection center, and the strain number was GDMCC 1.388.
a. Culturing seeds in a triangular flask: preparing a liquid culture medium, adjusting the pH value to 7, subpackaging into 2000ml triangular flasks, sterilizing at 121 ℃ for 20 minutes in each 1500ml triangular flask, cooling, inoculating the preserved enterococcus faecium into the triangular flask, culturing for 20 hours at 38 ℃ under an anaerobic condition, transferring the grown enterococcus faecium seeds into the 2000ml triangular flask according to the inoculation amount of 10%, and performing static culture for 20 hours at 37 ℃.
b. Seed tank culture: adding a liquid culture medium into a seeding tank, wherein the liquid loading amount is 70%, adjusting the pH value to 7, stirring, sterilizing at 121 ℃ for 30 minutes, cooling with cooling water, introducing sterile nitrogen to control the tank pressure to be 0.05MPa, transferring the grown 2000ml of triangular flask seed liquid into the liquid culture medium of the seeding tank at 10% of the inoculation amount when the temperature is reduced to 37 ℃, keeping the temperature at 37 ℃ and the tank pressure to be 0.05MPa, and statically culturing for 20 hours, wherein the pH value of the enterococcus faecium seed liquid is 4.5, and the bacterial count reaches 5-15 hundred million cfu/ml.
The formula of the liquid culture medium is as follows: 15g of peptone, 3g of glucose, 4g of corn flour, 8g of soybean meal, 4g of yeast extract, 0.5g of monopotassium phosphate, 0.6g of magnesium sulfate, 15mg of manganese sulfate and 1000ml of water.
Example 3
Preparation of composite fermentation liquor
Transferring enterococcus faecium seed liquid into a fermentation tank at an inoculation amount of 10%, performing static culture at 30-38 deg.C and 0.04-0.06MPa until enterococcus faecium reaches logarithmic phase;
b, adjusting the pH value in the fermentation tank to 7.2; adding 5g/L calcium carbonate and 1g/L dipotassium hydrogen phosphate;
c, transferring the bacillus coagulans seed solution into a fermentation tank according to the inoculation amount of 10%, maintaining the temperature at 37 ℃, the pressure at 0.05MPa and the rotation/minute at 120, and carrying out submerged aeration and stirring culture for 26 hours.
When the spore rate of the bacillus coagulans in the fermentation tank liquid reaches 90-95%, centrifuging, and collecting concentrated bacterial liquid and fermentation supernatant; drying the concentrated bacterial liquid and then using the dried concentrated bacterial liquid to prepare a feed additive; adjusting the pH value of the fermentation supernatant to 3.5-4.5 by using hydrochloric acid to obtain the composite fermentation liquor.
Example 4
The embodiment of the invention discloses an aquaculture antidote, which comprises the following raw materials in parts by weight: 5% of active carbon, 10% of biological enzyme, 30% of diatomite, 15% of antibacterial peptide and the balance of composite fermentation liquor; wherein the composite fermentation liquid is the supernatant liquid obtained by mixing and fermenting bacillus coagulans and enterococcus faecium.
Example 5
The embodiment of the invention discloses an aquaculture antidote, which comprises the following raw materials in parts by weight: 10% of active carbon, 15% of biological enzyme, 20% of diatomite, 10% of antibacterial peptide and the balance of composite fermentation liquor; wherein the composite fermentation liquid is the supernatant liquid obtained by mixing and fermenting bacillus coagulans and enterococcus faecium.
Example 6
The embodiment of the invention discloses an aquaculture antidote, which comprises the following raw materials in parts by weight: 15% of active carbon, 5% of biological enzyme, 25% of diatomite, 10% of antibacterial peptide and the balance of composite fermentation liquor; wherein the composite fermentation liquid is the supernatant liquid obtained by mixing and fermenting bacillus coagulans and enterococcus faecium.
Example 7
The embodiment of the invention discloses an aquaculture antidote, which comprises the following raw materials in parts by weight: 10% of activated carbon, 12% of biological enzyme, 26% of diatomite, 16% of antibacterial peptide and the balance of composite fermentation liquor; wherein the composite fermentation liquid is the supernatant liquid obtained by mixing and fermenting bacillus coagulans and enterococcus faecium.
Comparative example 1
The embodiment of the invention discloses an aquaculture antidote, which comprises the following raw materials in parts by weight: 4% of active carbon, 20% of biological enzyme, 35% of diatomite, 10% of antibacterial peptide and the balance of composite fermentation liquor; wherein the composite fermentation liquid is the supernatant liquid obtained by mixing and fermenting bacillus coagulans and enterococcus faecium.
Example 8
The results of monitoring the pH, dissolved oxygen, the number of pathogenic microorganisms, Chemical Oxygen Demand (COD), and transparency of the water body after application of the aquaculture antidotes of examples 4 to 7 and comparative example 1 are shown in table 1. Wherein, the sampling of the water body is to randomly select the water quality 0.5m away from the bank in the pond for cultivating the fish and the shrimp for detection, and each index is detected by adopting the detection method in the prior art.
TABLE 1 Effect table after administration of different aquaculture antidotes
Note: wherein "-" in Table 1 means that no treatment was performed.
As can be seen from Table 1, the aquaculture antidotes of the above examples 4-7 have the functions of stabilizing pH of water, increasing nutrition of water, killing pathogenic microorganisms and purifying water.
The invention discloses an aquaculture antidote which has the characteristics of no pollution, no toxicity, good stability, high safety performance, high antibacterial efficiency and the like. In addition, the antidote also has the performance of increasing the dissolved oxygen in water, improving water quality, absorbing and degrading organic matters in water and effectively sterilizing.
The embodiments in the present description are described in a progressive manner, each embodiment focuses on differences from other embodiments, and the same and similar parts among the embodiments are referred to each other. The device disclosed by the embodiment corresponds to the method disclosed by the embodiment, so that the description is simple, and the relevant points can be referred to the method part for description.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (1)
1. An aquaculture antidote is characterized by comprising the following raw materials in parts by weight: 5-15% of active carbon, 5-15% of biological enzyme, 20-30% of diatomite, 10-20% of antibacterial peptide and the balance of composite fermentation liquor; wherein the composite fermentation liquid is a supernatant liquid obtained by mixing and fermenting bacillus coagulans and enterococcus faecium;
the preparation method of the composite fermentation liquor comprises the following steps:
(1) culturing bacillus coagulans to generate bacillus coagulans seed liquid;
(2) culturing enterococcus faecium to generate enterococcus faecium seed liquid;
(3) inoculating the bacillus coagulans seed liquid and the enterococcus faecium seed liquid into the same fermentation tank for mixed fermentation;
(4) preparing composite fermentation liquor;
the preparation method of the bacillus coagulans seed liquid in the step (1) comprises the following steps:
transferring the preserved bacillus coagulans to a freshly prepared solid culture medium, culturing for 24 hours at 30-37 ℃, transferring the slant strain to an eggplant bottle slant, culturing for 24 hours at 30-37 ℃, cleaning the solid culture medium, and collecting lawn washing liquid; the components of the solid culture medium comprise: 8-10g of peptone, 5-7g of beef extract, 5-7g of sodium chloride and 18-20g of agar, fixing the volume to 1000ml by using distilled water, and adjusting the pH value to 7.2-7.4;
b, inoculating the lawn washing liquid obtained in the step a into a liquid culture medium in a seed tank, carrying out deep aeration stirring culture for 16-24 hours at the temperature of 30-38 ℃ and at the pressure of 0.04-0.06MPa and at the speed of 120-; the components of the liquid culture medium comprise: 8-20g of peptone, 5-15g of yeast extract, 0.2-2g of dipotassium phosphate, 0.2-2g of monopotassium phosphate, 0.1-2g of magnesium sulfate, 8-20g of glucose, 1-10g of calcium carbonate and 1000ml of water;
the preparation method of the enterococcus faecium seed liquid in the step (2) comprises the following steps:
inoculating enterococcus faecium to a liquid culture medium in a triangular flask, culturing for 14-24 hours at 30-38 ℃ under anaerobic conditions, transferring the grown enterococcus faecium liquid into another triangular flask liquid culture medium with the inoculation amount of 5-10%, and statically culturing for 14-24 hours at 30-38 ℃; the components of the liquid culture medium in the step (2) comprise: 5-15g of peptone, 0.5-5g of glucose, 4-6g of corn meal, 4-10g of soybean meal, 4-6g of yeast extract, 0.5-1.5g of monopotassium phosphate, 0.3-1g of magnesium sulfate, 5-20mg of manganese sulfate and 1000ml of water;
b, transferring the other triangular flask bacterial liquid into a liquid culture medium of a seeding tank according to the inoculation amount of 5-10%, and statically culturing for 14-24 hours at 30-38 ℃ and 0.04-0.06MPa to obtain the enterococcus faecium seed liquid; wherein the pH value of the enterococcus faecium seed liquid is 4.5-5.0, and the bacterial count reaches 5-15 hundred million cfu/ml;
the specific steps of the step (3) are as follows:
transferring the enterococcus faecium seed liquid into a fermentation tank by an inoculation amount of 5-10%, and statically culturing at 30-38 ℃ and 0.04-0.06MPa until the enterococcus faecium reaches the logarithmic phase;
b, adjusting the pH value in the fermentation tank to 7.2-7.4; adding 1-10g/L calcium carbonate and 0.2-2g/L dipotassium hydrogen phosphate;
c, transferring the bacillus coagulans seed solution into a fermentation tank with the inoculation amount of 5-10%, keeping the temperature at 30-38 ℃, the pressure at 0.04-0.06MPa and the pressure at 120-;
the specific steps of the step (4) are as follows:
centrifuging when the spore rate of the bacillus coagulans in the fermentation tank liquid reaches 90% -95%, and collecting concentrated bacterial liquid and fermentation supernatant; drying the concentrated bacterial liquid and then using the dried concentrated bacterial liquid to prepare a feed additive; and regulating the pH value of the fermentation supernatant to 3.5-4.5 by using hydrochloric acid to obtain the composite fermentation liquid.
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| CN107459204A (en) * | 2017-07-31 | 2017-12-12 | 浦江县酉泽水产科技有限公司 | The processing method of aquiculture waste water |
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| CN107459204A (en) * | 2017-07-31 | 2017-12-12 | 浦江县酉泽水产科技有限公司 | The processing method of aquiculture waste water |
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Denomination of invention: An antidote for aquaculture Effective date of registration: 20221115 Granted publication date: 20210316 Pledgee: Industrial Bank Co.,Ltd. Yancheng branch Pledgor: JIANGSU YUANSHAN BIOLOGY TECHNOLOGY CO.,LTD. Registration number: Y2022320000705 |