CN101698539B - Microecological agent for purifying water and preparation method thereof - Google Patents
Microecological agent for purifying water and preparation method thereof Download PDFInfo
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- CN101698539B CN101698539B CN2009102354141A CN200910235414A CN101698539B CN 101698539 B CN101698539 B CN 101698539B CN 2009102354141 A CN2009102354141 A CN 2009102354141A CN 200910235414 A CN200910235414 A CN 200910235414A CN 101698539 B CN101698539 B CN 101698539B
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- fungicides
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 96
- 238000002360 preparation method Methods 0.000 title claims abstract description 22
- 239000003795 chemical substances by application Substances 0.000 title abstract description 20
- 239000007787 solid Substances 0.000 claims abstract description 71
- 241000894006 Bacteria Species 0.000 claims abstract description 61
- 230000001580 bacterial effect Effects 0.000 claims abstract description 44
- 230000000813 microbial effect Effects 0.000 claims abstract description 35
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 26
- 230000000243 photosynthetic effect Effects 0.000 claims abstract description 19
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 12
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 12
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims abstract description 9
- 238000009360 aquaculture Methods 0.000 claims abstract description 9
- 244000144974 aquaculture Species 0.000 claims abstract description 9
- 239000000417 fungicide Substances 0.000 claims description 66
- 230000000855 fungicidal effect Effects 0.000 claims description 50
- 235000018291 probiotics Nutrition 0.000 claims description 38
- 239000006041 probiotic Substances 0.000 claims description 30
- 239000002245 particle Substances 0.000 claims description 25
- 239000000203 mixture Substances 0.000 claims description 15
- 239000000843 powder Substances 0.000 claims description 13
- 241000190950 Rhodopseudomonas palustris Species 0.000 claims description 12
- 238000003756 stirring Methods 0.000 claims description 12
- 241000187561 Rhodococcus erythropolis Species 0.000 claims description 8
- 244000005700 microbiome Species 0.000 claims description 8
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 7
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims description 7
- 229910021536 Zeolite Inorganic materials 0.000 claims description 7
- HNPSIPDUKPIQMN-UHFFFAOYSA-N dioxosilane;oxo(oxoalumanyloxy)alumane Chemical compound O=[Si]=O.O=[Al]O[Al]=O HNPSIPDUKPIQMN-UHFFFAOYSA-N 0.000 claims description 7
- 239000010457 zeolite Substances 0.000 claims description 7
- 238000010298 pulverizing process Methods 0.000 claims description 6
- 238000005469 granulation Methods 0.000 claims description 5
- 230000003179 granulation Effects 0.000 claims description 5
- 241000589614 Pseudomonas stutzeri Species 0.000 claims description 4
- 241000191043 Rhodobacter sphaeroides Species 0.000 claims description 4
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 2
- 241000459479 Capsula Species 0.000 claims description 2
- 101100412856 Mus musculus Rhod gene Proteins 0.000 claims description 2
- 241000589516 Pseudomonas Species 0.000 claims description 2
- 241000589540 Pseudomonas fluorescens Species 0.000 claims description 2
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- 101100242191 Tetraodon nigroviridis rho gene Proteins 0.000 claims description 2
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- 241000251468 Actinopterygii Species 0.000 abstract description 14
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 abstract description 12
- 239000000126 substance Substances 0.000 abstract description 11
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 abstract description 9
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- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 6
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- 239000001301 oxygen Substances 0.000 abstract description 5
- 229910052760 oxygen Inorganic materials 0.000 abstract description 5
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- 239000011574 phosphorus Substances 0.000 abstract description 4
- UCKMPCXJQFINFW-UHFFFAOYSA-N Sulphide Chemical compound [S-2] UCKMPCXJQFINFW-UHFFFAOYSA-N 0.000 abstract description 3
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- JMANVNJQNLATNU-UHFFFAOYSA-N oxalonitrile Chemical compound N#CC#N JMANVNJQNLATNU-UHFFFAOYSA-N 0.000 description 6
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- 239000000230 xanthan gum Substances 0.000 description 6
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- 241000276707 Tilapia Species 0.000 description 5
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- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 4
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- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 2
- PLXBWHJQWKZRKG-UHFFFAOYSA-N Resazurin Chemical compound C1=CC(=O)C=C2OC3=CC(O)=CC=C3[N+]([O-])=C21 PLXBWHJQWKZRKG-UHFFFAOYSA-N 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical class [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 241001494106 Stenotomus chrysops Species 0.000 description 2
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 description 2
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- 230000007096 poisonous effect Effects 0.000 description 2
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- LJCNRYVRMXRIQR-OLXYHTOASA-L potassium sodium L-tartrate Chemical compound [Na+].[K+].[O-]C(=O)[C@H](O)[C@@H](O)C([O-])=O LJCNRYVRMXRIQR-OLXYHTOASA-L 0.000 description 2
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- 235000011091 sodium acetates Nutrition 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
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- PODWXQQNRWNDGD-UHFFFAOYSA-L sodium thiosulfate pentahydrate Chemical compound O.O.O.O.O.[Na+].[Na+].[O-]S([S-])(=O)=O PODWXQQNRWNDGD-UHFFFAOYSA-L 0.000 description 2
- 239000003440 toxic substance Substances 0.000 description 2
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Landscapes
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
- Separation Of Suspended Particles By Flocculating Agents (AREA)
Abstract
The invention discloses a microecological agent for purifying water and a preparation method thereof. The microecological agent consists of the following components in part by weight: 13 to 17 parts of solid bacterial reagent of photosynthetic bacteria, 12.5 to 16.5 parts of solid bacterial reagent of bacillus subtilis, 10.5 to 13.3 parts of solid bacterial reagent of Saccharomyces cerevisiae, 33 to 37 parts of solid bacterial reagent of denitrifying bacteria and 0.3 to 0.7 part of microbial flocculant. The microecological agent can increase the transparency and dissolved oxygen in water, degrades, absorbs and converts organic pollutants in aquaculture environment and harmful substances such as nitrite, sulphide, nitrogen, phosphorus and the like in the water, ensures that the whole culture period does not need changing water, and can reduce the content of the harmful substances such as ammonia nitrogen, nitrite and the like in the water effectively, reduce feed coefficient, decrease production cost, quickly optimize the culture environment, strengthen capacity of resisting diseases of cultured fish, and promote the quick growth of cultured objects.
Description
Technical field
The present invention relates to a kind of water body purification agent, relate in particular to probiotics of a kind of purifying water body and preparation method thereof, the invention still further relates to this probiotics in Application for Field such as purifying aquatic product cultivating water body, water body reparation, water body environment improvement, belong to the water body purification field.
Background technology
At present, the water resource environment of China is seriously being perplexed in go from bad to worse and the pollution problem of water body environment, and the body eutrophication that nitrogen, phosphorus cause makes the objectionable impurities accumulation of aquaculture water, has endangered the aquaculture water environment; The organic waste water of manufacturing district discharging contains toxic substances such as phenol, cyanogen and has aggravated pollution level of water body environment or the like.Country is along with the rise and the development of microorganism water technology, and the pollution of development and utilization new microbial preparation improvement water body environment has been subjected to people's common concern.
Summary of the invention
One of purpose of the present invention is to overcome the deficiencies in the prior art, and the probiotics of a kind of effective purifying water body, enhancing aquatic products object resistance against diseases is provided;
Two of purpose of the present invention provides a kind of method for preparing above-mentioned probiotics;
Three of purpose of the present invention is with fields such as the purification of above-mentioned probiotics water body and reparation, water body environment improvement;
Above-mentioned purpose of the present invention is achieved through the following technical solutions:
The probiotics of a kind of purifying water body, enhancing aquaculture object resistance against diseases, each component that comprises following weight part is formed: 13～17 parts of photosynthetic bacterium solid fungicides, 12.5～16.5 parts of subtilis (Bacillus subtilis) solid fungicides, 10.5～13.3 parts of S. cervisiae (Saccharomyces cerevisiae) solid fungicides, 0.3～0.7 part of 33～37 parts of denitrifying bacterium solid fungicides and microbial flocculant;
Preferably, the weight part of each component is: 15.5 parts of photosynthetic bacterium solid fungicides, 12.5 parts of subtilis (Bacillus subtilis) solid fungicides, 12 parts of S. cervisiae (Saccharomyces cerevisiae) solid fungicides, 36 parts of denitrifying bacterium solid fungicides, 0.4 part of microbial flocculant;
Described photosynthetic bacterium is selected from any one in Rhodopseudomonas palustris (Rhodopseudomonaspalustris), capsula Rhodopseudomonas (Rhodopseudomonas capsulatus) or the Rhodopseudomonas spheroides (Rhodopseudomonas sphaeroides);
Described denitrifying bacterium is selected from any one in the false monospore bacillus (Pseudomonasstutzeri) of denitrification, denitrification thiobacillus (Thiobacillus denitrificans) or the fluorescent pseudomonas (Pseudomonas fluorescence);
Microbial flocculant (Microbial Flocculant, MBF) be that the microorganism of some kind is under the certain culture condition, the material that growth metabolism produces to certain phase with flocculation activity, by microbial fermentation, extraction, refining a kind of efficient, the nontoxic material for water treatment that obtains, generally form by polymer substances such as polysaccharide, protein, Mierocrystalline cellulose, glycoprotein and polyamino acid, contain multiple functional group in the molecule, can make the mutual coagulation of water-borne glue body suspended substance phase.
In order to reach better effect, microbial flocculant of the present invention is preferably and adopts Rhod microorganism (Rhodococcus etythropolis) to prepare the flocculation agent that obtains, for example, the rhodococcus erythropolis inoculation that purchase is obtained is cultivated on the seed culture and is obtained inoculum; Inoculum is inoculated in the fermention medium in the fermentor tank ferments, the sorbent material of back adding weighting agent or pulverizing is finished in fermentation, obtains solid fungicide, promptly.
Preferred, the flocculation agent NOC-1 that the present invention adopts rhodococcus erythropolis (Rhodococcus erythropolis) S-1 bacterial strain to make, this flocculation agent NOC-1 (for example: the new golden dye chemical industry in Baoding company limited can buy by commercial sources and obtain, trade name: microbial flocculant NOC-1, specification: molecular weight 70-100 ten thousand), also can prepare (Kurane R according to the disclosed method of document, et al.Culture Conditions for Production of MicrobialFlocculant by Rhodacus Ergthropolis[J] .Agric.Biol.Chem, 1986,50 (9): 2309-2313.)
Subtilis solid fungicide of the present invention, S. cervisiae solid fungicide, denitrifying bacterium solid fungicide or photosynthetic bacterium solid fungicide can prepare solid fungicide according to following ordinary method: described bacterial classification inoculation is cultivated on the seed culture obtained inoculum; Inoculum is inoculated in the fermention medium in the fermentor tank ferments, the fermented liquid after fermentation is finished is adsorbed on the sorbent material of being made up of organic mixture, promptly obtains corresponding solid fungicide.
Wherein, described photosynthetic bacterium solid fungicide can prepare with reference to following method:
Rhodopseudomonas palustris (Rhodopseudomonas palustris) (Guangdong Microbes Inst culture presevation administrative center, goods number: the GIM1.167) preparation of microbial inoculum: the formulated substratum that provides by supplier, the Rhodopseudomonas palustris bacterial classification inoculation is cultivated in substratum, medium slant seed bacterial strain added behind the mineral oil store in 4 ℃ of refrigerators, take out twice of activation before using, change the bacterial strain after the activation over to 500 milliliters of vial illumination liquid culture, culture temperature is 30 ℃, cultivates 96 hours.Change 5L vial illumination cultivation 96 hours afterwards over to as secondary seed solution.Add substratum in 1 ton fermentor tank, it consists of: 1.145 kilograms of sodium-acetates, 0.055 kilogram of peptone, 0.6 kilogram of sodium bicarbonate, 0.4 kilogram in Sulfothiorine, 0.3 kilogram in sodium-chlor, 0.1 kilogram in sal epsom, 0.05 kilogram of potassium primary phosphate.With filtering 0.1N phosphoric acid adjust pH 7.0.Illumination 30001x.To producing a jar inoculation, inoculum size is 50kg, begins fermentation from seeding tank.Fermentation condition: culture temperature is 30 ℃, inoculate after 20 hours and took a sample once, observe the thalli growth situation, whether pollution arranged, after liquid fermenting is finished every 4 hours, in zymocyte liquid, add conventional sorbent material and weighting agent xanthan gum, get the Rhodopseudomonas palustris microbial inoculum.
Described subtilis solid fungicide can prepare with reference to following method:
Subtilis (Bacillus subtilis) (Chinese agriculture microbial strains preservation administrative center, goods number: the ACCC01031) preparation of microbial inoculum: the formulated substratum that provides by supplier, Bacillus subtilis strain is inoculated in the substratum cultivates, to store in 4 ℃ of refrigerators of medium slant seed bacterial strain, take out twice of activation before using, bacterial strain after the activation is changeed 250 milliliters of eggplant bottle nutrient agar mediums to be cultivated, culture temperature is 24 ℃, wash with the water bacterium of going out during 48 hours well-growns, as the inoculum of seeding tank.Add substratum in 1 ton fermentor tank, it consists of: Semen Maydis powder 3%, soybean cake powder 3%, bran powder 3%, K
2HPO
42.15%, KH
2PO
40.8%, MgSO
40.075%, peanut oil 0.3%.Initial pH7.5.To producing a jar inoculation, begin fermentation from seeding tank.Fermentation condition: culture temperature is 37 ℃, and ventilation is 1: 1.1, and tank pressure is 0.05mpa/cm
2, stirring velocity is 220rpm, inoculates after 20 hours and takes a sample once every 4 hours, observe the thalli growth situation, whether pollution arranged, wait to find spore generate reach more than 90% after, liquid fermenting is finished, in zymocyte liquid, add conventional sorbent material and weighting agent xanthan gum, get bacillus subtilis microbial agent.
Described S. cervisiae solid fungicide can prepare with reference to following method:
S. cervisiae (Saccharomyces cerevisiae) (Guangdong Microbes Inst culture presevation administrative center, goods number: GIM 2.39) preparation of microbial inoculum: the formulated substratum that provides by supplier, the saccharomyces cerevisiae bacterial classification inoculation is cultivated in substratum, medium slant seed bacterial strain added behind the mineral oil store in 4 ℃ of refrigerators, take out twice of activation before using, bacterial strain after the activation is changed in 250 milliliters of eggplant bottles and cultivates, culture temperature is 28-30 ℃, cultivated 48 hours, with sterile saline it is washed when strain growth is good, eluate is the inoculum of seeding tank.In 1 ton fermentor tank, add substratum, wort, 10～15 ripple woodss are transferred pH to 6.4.To producing a jar inoculation, inoculum size is 50kg, begins fermentation from seeding tank.Fermentation condition: culture temperature is 28 ℃, and tank pressure is 0.05mpa/cm
2, stirring velocity is 220rpm, ventilating ratio 1: 1.1, inoculate after 20 hours and took a sample once, observe the thalli growth situation, whether pollution arranged, after liquid fermenting is finished every 4 hours, in zymocyte liquid, add conventional sorbent material and weighting agent xanthan gum, get the cereuisiae fermentum bacteria agent.
Described denitrifying bacterium solid fungicide can prepare with reference to following method:
Denitrifying bacterium (Pseudomonas stutzeri) (Chinese industrial microbial strains preservation administrative center, goods number: CICC 23621) preparation of microbial inoculum: the formulated substratum that provides by supplier, the denitrifying bacterium bacterial classification inoculation is cultivated in substratum, medium slant seed bacterial strain added behind the mineral oil store in 4 ℃ of refrigerators, take out twice of activation before using, bacterial strain after the activation is changed in 250 milliliters of eggplant bottles and cultivates, culture temperature is 37 ℃, wash with aqua sterilisa during 36 hours well-growns, as the inoculum of seeding tank.Add substratum in 1 ton fermentor tank, it consists of: 2.0kg KNO
3, 2.0kg NaNO
3, 0.03kg MgSO
47H
2O, 0.5kg K
2HPO
4, 10kg Seignette salt, 1.0kg KH
2PO
4, 0.5kg FeCl
26H
2O, 0.2kg CaCl
22H
2O, 0.1L concentration are resazurin solution, the 0.5kg L-halfcystine of 0.2g/L, and the pH value is 7-7.5.To producing a jar inoculation, inoculum size is 5%, begins fermentation from seeding tank.Fermentation condition: culture temperature is 37 ℃, and ventilation is 1: 1.1, and tank pressure is 0.05mpa/cm
2, stirring velocity is 220rpm, and liquid fermenting is finished, and fermented liquid is adsorbed on the sorbent material of being made up of organic mixture (for example: zeolite powder 30-50 part, dregs of beans 35-40 part and pomace 5-15 part), and oven drying at low temperature gets the denitrifying bacterium solid fungicide.
Described microbial flocculant can prepare with reference to following method:
Rhodococcus erythropolis (Rhodococcus erythropolis) (Chinese common micro-organisms culture presevation administrative center, goods number: CGMCC 4.1491) preparation of microbial inoculum: the formulated substratum that provides by supplier, the rhodococcus erythropolis bacterial classification inoculation is cultivated in substratum, to store in 4 ℃ of refrigerators of medium slant seed bacterial strain, take out twice of activation before using, bacterial strain after the activation is changed in 250 milliliters of eggplant bottles and cultivates, culture temperature is 37 ℃, wash with aqua sterilisa during 36 hours well-growns, as the inoculum of seeding tank.Add substratum in 1 ton fermentor tank, it consists of: 2.43kgK
2HPO
4, 14.0kgNa
2HPO
412H
2O, 2.0kgNH
4Cl, 0.35kg CaCl
26H
2O, 0.001kgCaCl
22H
2O, 0.001kgFeCl
36H
2O, 0.004kgMnCl
24H
2O, glucose 10kg.To producing a jar inoculation, inoculum size is 5%, begins fermentation from seeding tank.Fermentation condition: culture temperature is 37 ℃, and tank pressure is 0.05mpa/cm
2Stirring velocity is 220rpm, and liquid fermenting is finished, and fermented liquid is adsorbed on the sorbent material of being made up of organic mixture (for example: zeolite powder 30-50 part, dregs of beans 35-40 part and pomace 5-15 part), oven drying at low temperature gets the rhodococcus erythropolis solid fungicide, gets microbial flocculant.
Used all microbial strainss of the present invention (can available from Guangdong Microbes Inst culture presevation administrative center, Chinese agriculture microbial strains preservation administrative center, Chinese common micro-organisms culture presevation administrative center, Chinese industrial microbial strains preservation administrative center) and other raw material all can be bought from market and obtain, and its specification meets national sector standard.
Another object of the present invention provides a kind of method for preparing the probiotics of above-mentioned purifying water body;
Another object of the present invention is to realize by the following technical programs:
A kind of method for preparing the probiotics of above-mentioned purifying water body may further comprise the steps:
(1) takes by weighing each component by described weight part;
(2) with the photosynthetic bacterium solid fungicide, the subtilis solid fungicide, the organic mixture of S. cervisiae solid fungicide, denitrifying bacterium solid fungicide and pulverizing mixes in proportion, and obtains mixed thing;
(3), obtain the former bacterial spawn shape of viable bacteria particle with the resulting mixed thing extruder grain of step (2); With the granulation of microbial flocculant disk, obtain spheroidal particle; Viable bacteria former bacterial spawn shape particle and spheroidal particle are mixed, stir, promptly.
Wherein, the organic mixture described in the step (2) preferably is made up of the component of following weight part: zeolite powder 30-50 part, dregs of beans 35-40 part and pomace 5-15 part; Add organic mixture amount can depend on the circumstances, add organic mixture weight account for the 10-35% of mixed thing gross weight, be preferably 23.6%;
The particulate of the former bacterial spawn shape of viable bacteria described in the step (3) diameter is preferably 0.3cm, and length is preferably 0.9～1.2cm; The diameter of described spheroidal particle is preferably 0.07cm.
The using method of probiotics of the present invention and consumption:
1, directly shed, every kilogram can be used for the 5-6 mu water surface (1 meter depth of water), and the high-density breeding pool is when substrate, can suitably increase consumption when water quality is relatively poor.
2, the breed initial stage uses once, cultures omnidistance every 1-2 month and uses once.
Photosynthetic bacterium in the probiotics of the present invention, subtilis (Bacillussubtilis) adaptability are strong, can restrain oneself the organic waste water of high depth and stronger decomposition and inversion ability, poisonous substances such as phenol, cyanogen there be stronger standing and capacity of decomposition, can reach and improve water body environment, recover water ecology equilibrated effect.Microbial flocculant can make the mutual coagulation of water-borne glue body suspended substance phase, effectively purifying water body, improve water quality, suppress the harmful microbe breeding.
Probiotics of the present invention makes full use of the anaerobism of relevant bacterial strain, good oxygen characteristic, strengthening aspect the purification of water quality of aquaculture water and the resistance against diseases performance synergy of aquatic products object, can increase water transparency and dissolved oxygen, organic pollutant (residual bait in degraded and absorbed and the conversion culture environment of aquatic products, ight soil and biological dead volume) and water body in nitrite, sulfide, nitrogen, objectionable impuritiess such as phosphorus, the whole culture-cycle need not be changed water, can reduce the content of harmful such as ammonia nitrogen ite of water body effectively, reduce feed coefficient, reduce production cost, can optimize breeding environment rapidly, strengthen the cultured fishes resistance against diseases, promote to culture the quick growth of object.
Concrete, the major advantage and the effect of probiotics of the present invention comprise the following aspects:
1, can reduce water surrounding and the pollution of soil property environment on every side, prevent the abuse of other pharmaceutical chemicals, can restrain oneself the organic waste water of high depth and stronger decomposition and inversion ability, poisonous substances such as phenol, cyanogen be had necessarily stand and capacity of decomposition, recover the water ecology balance.
2, improved the fishery water quality environment effectively, administered the eutrophication of water body, eliminated the ammonia and nitrogen pollution of water body, toxic substances such as the nitrite in the degraded water body, sulfide.Reduce discharging and change the waterside number, reduced medication and pollution, improved the value of aquaculture product itself, saved water resources.
3, in water body, can breed in a large number, effectively suppress harmful bacterium and algae grows, when purifying water, create good water ecological setting again.Simultaneously can serve as bait, as the prophylactic function of fodder additives.To making " green fishery " and " Sustainable development fishery " established the good technical basis.
4, probiotics of the present invention adopts microbial flocculant, has avoided the toxicity hazard of chemical floc such as polyacrylamide, has guaranteed the graininess of probiotics simultaneously, is difficult for being eaten by filter-feeding fish, has strengthened the effect of purifying water body.
Embodiment
Further describe the present invention below in conjunction with specific embodiment, advantage of the present invention and characteristics will be more clear along with description.But these embodiment only are exemplary, scope of the present invention are not constituted any restriction.It will be understood by those skilled in the art that and down can make amendment or replace without departing from the spirit and scope of the present invention, but these modifications and replacing all fall within the scope of protection of the present invention the details of technical solution of the present invention and form.
The preparation of embodiment 1 material
(1) preparation of Rhodopseudomonas palustris (Rhodopseudomonas palustris) microbial inoculum:
The formulated substratum that provides by supplier, the Rhodopseudomonas palustris bacterial classification inoculation is cultivated in substratum, medium slant seed bacterial strain added behind the mineral oil store in 4 ℃ of refrigerators, take out twice of activation before using, change the bacterial strain after the activation over to 500 milliliters of vial illumination liquid culture, culture temperature is 30 ℃, cultivates 96 hours.Change 5L vial illumination cultivation 96 hours afterwards over to as secondary seed solution.Add substratum in 1 ton fermentor tank, it consists of: 1.145 kilograms of sodium-acetates, 0.055 kilogram of peptone, 0.6 kilogram of sodium bicarbonate, 0.4 kilogram in Sulfothiorine, 0.3 kilogram in sodium-chlor, 0.1 kilogram in sal epsom, 0.05 kilogram of potassium primary phosphate.With filtering 0.1N phosphoric acid adjust pH 7.0.Illumination 30001x.To producing a jar inoculation, inoculum size is 50kg, begins fermentation from seeding tank.Fermentation condition: culture temperature is 30 ℃, inoculates after 20 hours and takes a sample once every 4 hours, observes the thalli growth situation, whether pollution is arranged, and after liquid fermenting is finished, adds sorbent material and weighting agent xanthan gum in zymocyte liquid, gets the Rhodopseudomonas palustris microbial inoculum.
(2) subtilis solid fungicide preparation:
Subtilis (Bacillus subtilis) (Chinese agriculture microbial strains preservation administrative center, goods number: the ACCC01031) preparation of microbial inoculum: the formulated substratum that provides by supplier, Bacillus subtilis strain is inoculated in the substratum cultivates, to store in 4 ℃ of refrigerators of medium slant seed bacterial strain, take out twice of activation before using, bacterial strain after the activation is changeed 250 milliliters of eggplant bottle nutrient agar mediums to be cultivated, culture temperature is 24 ℃, wash with the water bacterium of going out during 48 hours well-growns, as the inoculum of seeding tank.Add substratum in 1 ton fermentor tank, it consists of: Semen Maydis powder 3%, soybean cake powder 3%, bran powder 3%, K
2HPO
42.15%, KH
2PO
40.8%, MgSO
40.075%, peanut oil 0.3%.Initial pH7.5.To producing a jar inoculation, begin fermentation from seeding tank.Fermentation condition: culture temperature is 37 ℃, and ventilation is 1: 1.1, and tank pressure is 0.05mpa/cm
2, stirring velocity is 220rpm, inoculates after 20 hours and takes a sample once every 4 hours, observe the thalli growth situation, whether pollution arranged, wait to find spore generate reach more than 90% after, liquid fermenting is finished, in zymocyte liquid, add sorbent material and weighting agent xanthan gum, get bacillus subtilis microbial agent.
(3) S. cervisiae solid fungicide preparation:
S. cervisiae (Saccharomyces cerevisiae) (Guangdong Microbes Inst culture presevation administrative center, goods number: GIM 2.39) preparation of microbial inoculum: the formulated substratum that provides by supplier, the saccharomyces cerevisiae bacterial classification inoculation is cultivated in substratum, medium slant seed bacterial strain added behind the mineral oil store in 4 ℃ of refrigerators, take out twice of activation before using, bacterial strain after the activation is changed in 250 milliliters of eggplant bottles and cultivates, culture temperature is 28-30 ℃, cultivated 48 hours, with sterile saline it is washed when strain growth is good, eluate is the inoculum of seeding tank.In 1 ton fermentor tank, add substratum, wort, 10～15 ripple woodss are transferred pH to 6.4.To producing a jar inoculation, inoculum size is 50kg, begins fermentation from seeding tank.Fermentation condition: culture temperature is 28 ℃, and tank pressure is 0.05mpa/cm
2, stirring velocity is 220rpm, ventilating ratio 1: 1.1, inoculate after 20 hours and took a sample once, observe the thalli growth situation, whether pollution arranged, after liquid fermenting is finished every 4 hours, in zymocyte liquid, add sorbent material and weighting agent xanthan gum, get the cereuisiae fermentum bacteria agent.
(4) denitrifying bacterium solid fungicide preparation:
Denitrifying bacterium (Pseudomonas stutzeri) (Chinese industrial microbial strains preservation administrative center, goods number: CICC 23621) preparation of microbial inoculum: the formulated substratum that provides by supplier, the denitrifying bacterium bacterial classification inoculation is cultivated in substratum, medium slant seed bacterial strain added behind the mineral oil store in 4 ℃ of refrigerators, take out twice of activation before using, bacterial strain after the activation is changed in 250 milliliters of eggplant bottles and cultivates, culture temperature is 37 ℃, wash with aqua sterilisa during 36 hours well-growns, as the inoculum of seeding tank.Add substratum in 1 ton fermentor tank, it consists of: 2.0kg KNO
3, 2.0kg NaNO
3, 0.03kg MgSO
47H
2O, 0.5kg K
2HPO
4, 10kg Seignette salt, 1.0kg KH
2PO
4, 0.5kg FeCl
26H
2O, 0.2kg CaCl
22H
2O, 0.1L concentration are resazurin solution, the 0.5kg L-halfcystine of 0.2g/L, and the pH value is 7-7.5.To producing a jar inoculation, inoculum size is 5%, begins fermentation from seeding tank.Fermentation condition: culture temperature is 37 ℃, and ventilation is 1: 1.1, and tank pressure is 0.05mpa/cm
2, stirring velocity is 220rpm, and liquid fermenting is finished, and fermented liquid is adsorbed on the sorbent material of being made up of organic mixture, and oven drying at low temperature gets the denitrifying bacterium solid fungicide.
(5), microbial flocculant is available from the new golden dye chemical industry in Baoding company limited, trade name: microbial flocculant NOC-1, specification: molecular weight 70-100 ten thousand (solid).
The preparation of embodiment 2 probioticses
(1) take by weighing each component by following weight:
Subtilis solid fungicide: 125g
S. cervisiae solid fungicide: 120g
Denitrifying bacterium solid fungicide: 360g
Photosynthetic bacterium solid fungicide: 155g
Microbial flocculant: 4.0g
(2) with the photosynthetic bacterium solid fungicide, the subtilis solid fungicide, the organic mixture of S. cervisiae solid fungicide and denitrifying bacterium solid fungicide and pulverizing (component by following weight part is formed: zeolite powder 30-50 part, dregs of beans 35-40 part, pomace 5-15 part) 307g mixes in proportion the back extruder grain, obtain the former bacterial spawn shape of viable bacteria particle, the particulate diameter is 0.3cm, and length is 0.9～1.2cm; With the granulation of microbial flocculant disk, obtain white spheroidal particle, the diameter of spheroidal particle is preferably 0.07cm; Viable bacteria former bacterial spawn shape particle and spheroidal particle are mixed, stir, promptly.
The preparation of embodiment 3 probioticses
(1) take by weighing each component by following weight:
Subtilis solid fungicide: 125g
S. cervisiae solid fungicide: 105g
Denitrifying bacterium solid fungicide: 330g
Photosynthetic bacterium solid fungicide: 130g
Microbial flocculant: 3.0g
(2) with the photosynthetic bacterium solid fungicide, the subtilis solid fungicide, the organic mixture of S. cervisiae solid fungicide and denitrifying bacterium solid fungicide and pulverizing (component by following weight part is formed: zeolite powder 30-50 part, dregs of beans 35-40 part, pomace 5-15 part) 235.5g mixes in proportion the back extruder grain, obtain the former bacterial spawn shape of viable bacteria particle, the particulate diameter is 0.3cm, and length is 0.9～1.2cm; With the granulation of microbial flocculant disk, obtain white spheroidal particle, the diameter of spheroidal particle is preferably 0.07cm; Viable bacteria former bacterial spawn shape particle and spheroidal particle are mixed, stir, promptly.
The preparation of embodiment 4 probioticses
(1) take by weighing each component by following weight:
Subtilis solid fungicide: 165g
S. cervisiae solid fungicide: 133g
Denitrifying bacterium solid fungicide: 370g
Photosynthetic bacterium solid fungicide: 170g
Microbial flocculant: 7.0g
(2) with the photosynthetic bacterium solid fungicide, the subtilis solid fungicide, the organic mixture of S. cervisiae solid fungicide and denitrifying bacterium solid fungicide and pulverizing (component by following weight part is formed: zeolite powder 30-50 part, dregs of beans 35-40 part, pomace 5-15 part) 155g mixes in proportion the back extruder grain, obtain the former bacterial spawn shape of viable bacteria particle, the particulate diameter is 0.3cm, and length is 0.9～1.2cm; With the granulation of microbial flocculant disk, obtain white spheroidal particle, the diameter of spheroidal particle is preferably 0.07cm; Viable bacteria former bacterial spawn shape particle and spheroidal particle are mixed, stir, promptly.
The effect test of test example 1 probiotics of the present invention
Industrialized culture workshop, Chaoyang District, Beijing City aquaculture service centre aquatic science and technology garden is to utilize water treating equipment, recycle water body, carry out tilapia, blue shark, the breed of surviving the winter of rainbow porgy, the workshop is divided into cultures workshop and two parts in water treatment workshop, wherein culture 1400 square metres of workshop areas, 400 square metres of water treatment workshop areas.
Culture the workshop and have 40 ponds, each pond is meter (length) * 1.8,2.8 meters (wide) * 8.0 meter (deeply), 35.8 tons of average retainings.500 square metres of water treatment workshop floor spaces.
All fishponds are total to 1432 tons of retainings, are total to 800 tons of retainings in the water treatment workshop in each biological filter, and retaining is about 30 tons in the equalizing tank, and 2262 tons in water body is arranged altogether.The workshop recycles two 10 kilowatts water pump, and per hour circulating water flow is 150 tons, and be 3600 tons every day, promptly circulates every day 1.6 times.Thermal well a glance is arranged in the workshop, be used for regulating water temperature about 22 degree.
The batch production workshop September 20 began test, fished for tilapia, blue shark, rainbow porgy that outdoor ponds is raised, dropped into the batch production workshop and raised, up to May in next year.12391 kilograms of fishes are dropped in the workshop altogether.
This experiment is probiotics of the present invention (probiotics that embodiment 2-4 is prepared) influence to fish growth and water quality under the industrialized culture condition, October 9 rose, with No. 10 ponds is control group (not adopting any water body purification measure), with No. 18, No. 19 ponds are test group, launch test.Content of the test is, with probiotics of the present invention respectively at 9:00,10:30,13:00,16:00 drop into 1kg in No. 18 and No. 19 ponds; No. 10 any water body purification measure is not adopted in the pond.No. 10 ponds, No. 18 ponds, No. 19 ponds self-control expanded pellet feed of all throwing something and feeding, the feeding volume of each pond every day is 50 kilograms.
After the experiment beginning, carry out the successive monitoring for water quality.The project of monitoring comprises pH value, temperature, ammonia one nitrogen, nitrous acid-nitrogen.
Master data when two, experiment begins:
Test is an experimental group with No. 18 ponds, No. 19 ponds, and the basic condition that No. 10 ponds are control group, three fishponds is as follows: water examination was accomplished December 7 totally 60 days continuously from October 9.The experimental session water temperature is between 21.5 to 23.0.
Take a sample respectively in November 9 and December 9 and on January 9th, 1, the fish growth situation that obtains sees Table 1:
Table 1 growing state
| October 9 | 11-9 | 12-9 | 1-9 | |
| Kind | Individual weight (g) | Individual weight (g) | Individual weight (g) | Individual weight (g) |
| Luo Fei | 44 | 95 | 156 | 215 |
| Blue shark | 125 | 180 | 202 | 240 |
| Luo Fei | 80 | 275 | 365 | 445 |
| Blue shark | 140 | 230 | 285 | 376 |
| Luo Fei | 80 | 115 | 250 | 321 |
| Blue shark | 140 | 195 | 225 | 268 |
From October 9 to 9, second on January, 50 of the dead tilapias in No. 10 ponds, 38 of blue sharks.32 of the dead tilapias in No. 19 ponds, 18 of blue sharks.35 of the dead tilapias in No. 18 ponds, 21 of blue sharks.
Three, test results and analysis
When 1, experiment began, the ammonia-nitrogen content of test group was lower than control group, but after the 18th day, the ammonia nitrogen of test group exceeds control group on the contrary, and before and after the 36th day, peaked.
2. the Changing Pattern of nitrous acid-nitrogen content in test group and the control group water quality:
No matter be control group, or test group, the variation of nitrous acid-nitrogen does not all have any definite rule.And between the Changing Pattern of ammonia nitrogen, do not see the trend what unanimity is arranged yet.
3. test group and control group growth change rule:
From the situation of fish growth, the growth of test group is significantly better than control group, and the amount of interpolation probiotics of the present invention the more, and growing state also better.In addition, from experimental session fish kills quantity, the dead quantity of test group fish the also dead quantity than control group fish is obviously low.
The effect test of test example 2 probioticses of the present invention
One, profile test
The experiment and demonstration project of fishery cultivating water quality purification techniques is carried out in this test in the Daxing District.Mainly be under the prerequisite of not changing water, by probiotics degraded and absorbed of the present invention with transform organic pollutant (residual bait, ight soil and biological dead volume) in the culture environment of aquatic products and nitrogen, phosphorus etc.And the dominant population of formation (beneficial microorganism) itself, thereby effectively suppress harmful microorganism and algae grows breeding.Reach the optimization breeding environment, strengthen the cultured fishes resistance against diseases, promote growth fast to wait net effect to test.
Two, test materials and method
1, test water body: pond fish and breeding water.
2, subjects: probiotics of the present invention (embodiment 2-4 is prepared).
3, test site: Cao Cun fishing ground, town, high official position shop, Daxing District.
4, test grouping: choose this water surface area and be all 12 mu, the depth of water and be all adjacent four ponds of 2 meters, CK pond (not adopting any water body purification measure), K1 pond in contrast uses photosynthetic bacterium with strong water treasured, K2 pond with probiotics of the present invention, K3 pond.
Transparency, the ammonia nitrogen of four groups measured at the every interval of duration of test 15 days, and index such as dissolved oxygen, nitrite, pH value and water temperature also writes down the result.The growing state of duration of test selective examination fish calculates average cabrage, gross weight and feed coefficient during end.
7, detection method: water quality detection is the main water quality analysis box that is used with the laboratory.First time of application April 15,, use altogether 13 times by the end of October 31.Test result is seen attached list.
Three, test results and analysis
1, cultivating pool test results and analysis
Can draw from testing data, the K2 pond is compared with the K3 pond with K1, ammonia nitrogen on average reduces by 37%, nitrite on average reduces by 46%, DO increases by 16.4%, and transparency improves 20%, pH value no significant difference, prove that fully probiotics of the present invention is obvious to improving the water quality effect, and whole structure to obviously be better than existing water body purification agent.
According to going out pool situation, the feed coefficient in K1 and K3 pond is 2.24, and the feed coefficient in K2 pond is 1.93, and feed coefficient has reduced by 13.8%.The yield rate in K1 and K3 pond is 87%, and test tank (K2 pond) yield rate is 96%, and the fishery product quality has improved 9%.
Four, conclusion (of pressure testing)
Remarkable by testing probiotics net effect of the present invention as can be seen, mainly show the following aspects:
1, forms dominant population, suppress the breeding of harmful bacterium.
2, absorb objectionable impurities such as ammonia nitrogen in the water body, improve breeding environment.
3, improve fish immunity power, reduce fish disease and take place.
4, promoting digestion absorbs, and increase can utilize bait quantity.
Test-results proves: probiotics of the present invention can increase water transparency and dissolved oxygen, the whole culture-cycle need not be changed water, can reduce the content of harmful such as ammonia nitrogen ite of water body when particularly water temperature is more than 25 ℃ effectively, reduce feed coefficient, reduce production cost, increase economic efficiency.Topmost effect is and can effectively improves water quality, and the back drug residue free is used in water saving, helps environment protection, meets the needs that modern water industry is nuisanceless, green, healthy aquaculture develops.
Claims (8)
1. the probiotics of a purifying water body, each component that comprises following weight part is formed: 13～17 parts of photosynthetic bacterium solid fungicides, 12.5～16.5 parts of subtilis (Bacillus subtilis) solid fungicides, 10.5～13.3 parts of S. cervisiae (Saccharomyces cerevisiae) solid fungicides, 0.3～0.7 part of 33～37 parts of denitrifying bacterium solid fungicides and microbial flocculant;
Its preparation method comprises: (1), with the photosynthetic bacterium solid fungicide, the subtilis solid fungicide, the organic mixture of S. cervisiae solid fungicide, denitrifying bacterium solid fungicide and pulverizing mixes in proportion, and obtains mixed thing; Described organic mixture is made up of the component of following weight part: zeolite powder 30-50 part, dregs of beans 35-40 part and pomace 5-15 part; Add organic mixture weight account for the 10-35% of mixed thing gross weight;
(2), with the resulting mixed thing extruder grain of step (1), obtain the former bacterial spawn shape of viable bacteria particle; With the granulation of microbial flocculant disk, obtain spheroidal particle; Viable bacteria former bacterial spawn shape particle and spheroidal particle are mixed, stir, promptly.
2. according to the described probiotics of claim 1, it is characterized in that, the weight part of each component is: 15.5 parts of photosynthetic bacterium solid fungicides, 12.5 parts of subtilis (Bacillussubtilis) solid fungicides, 12 parts of S. cervisiae (Saccharomyces cerevisiae) solid fungicides, 36 parts of denitrifying bacterium solid fungicides, 0.4 part of microbial flocculant.
3. according to claim 1 or 2 described probioticses, it is characterized in that: described photosynthetic bacterium is selected from any one in Rhodopseudomonas palustris (Rhodopseudomonas palustris), capsula Rhodopseudomonas (Rhodopseudomonas capsulatus) or the Rhodopseudomonas spheroides (Rhodopseudomonas sphaeroides).
4. according to claim 1 or 2 described probioticses, it is characterized in that: described denitrifying bacterium is selected from any one in the false monospore bacillus (Pseudomonas stutzeri) of denitrification, denitrification thiobacillus (Thiobacillus denitrificans) or the fluorescent pseudomonas (Pseudomonasfluorescence).
5. according to claim 1 or 2 described probioticses, it is characterized in that: described microbial flocculant is the solid fungicide that adopts Rhod microorganism (Rhodococcus erythropolis) to prepare.
6. according to the described probiotics of claim 1, it is characterized in that: the weight that step (1) adds organic mixture accounts for 23.6% of mixed thing gross weight.
7. according to the described probiotics of claim 1, it is characterized in that: the particulate of the former bacterial spawn shape of viable bacteria described in the step (2) diameter is 0.3cm, and length is 0.9～1.2cm; The diameter of described spheroidal particle is 0.07cm.
8. claim 1 or the 2 described probioticses application in purifying water body, the growth of promotion aquaculture object.
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| CN108545829B (en) * | 2018-04-25 | 2021-08-17 | 中国科学院烟台海岸带研究所 | Method for in-situ aerobic denitrification treatment and purification of aquaculture sewage |
| CN110655202B (en) * | 2018-12-25 | 2020-08-04 | 四川千路环保科技有限责任公司 | Artificial capsule type multifunctional particle and preparation method and application thereof |
| CN110283740B (en) * | 2019-05-31 | 2021-01-12 | 新疆天康饲料科技有限公司生物添加剂分公司 | Compound bacterium agent for degrading high ammonia nitrogen sewage and application of compound bacterium agent in sewage treatment |
| CN111657207A (en) * | 2020-05-22 | 2020-09-15 | 太原市邦侬农业发展有限公司 | Fish and vegetable symbiotic system and special photosynthetic bacteria agent for aquaculture |
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2009
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