CN102649936A - Compound microorganism fungicide for improving water quality of culturing water body and preparation method - Google Patents
Compound microorganism fungicide for improving water quality of culturing water body and preparation method Download PDFInfo
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Abstract
The invention discloses a compound microorganism fungicide for improving the water quality of a culturing water body and a preparation method. The compound microorganism fungicide is prepared by taking the strains by weight percent: bacillus subtilis, yeast, lactic acid bacteria, photosynthetic bacteria, nitrifying bacteria, bacillus licheniformis, and the like. The preparation method comprises the following steps of: firstly, culturing the bacillus subtilis and bacillus licheniformis: culturing the bacillus subtilis and bacillus licheniformis on a beef extract peptone culture medium; secondly, culturing the yeast: culturing the yeast on a lactic acid-potato-glucose culture medium; thirdly, culturing the lactic acid bacteria: culturing the lactic acid bacteria on a wort culture medium, inoculating on a triangular flask for culturing, and finally inoculating on a solid culture medium for culturing; fourthly, culturing the photosynthetic bacteria; fifthly, culturing the nitrifying bacteria; and sixthly, stirring and mixing the cultured strains by weight ratio, and drying at low temperature, thereby obtaining the compound microorganism fungicide. The formula is reasonable; the use is convenient; water is uniformly splashed; the method is easily performed; and the operation is convenient.
Description
Technical field
The invention belongs to microbial technology field; More specifically relate to a kind of complex microorganism preparations that is used for the aquaculture water water quality improvement; The preparation method who also relates to this biotechnological formulation simultaneously, said preparation is applicable to the breed of fingerling and adult fish in the aquaculture wateies such as lake, reservoir, pond.
Background technology
Development along with culture fishery; The batch production high-density breeding needs a large amount of feeds that drop into; The remaining movement with aquatic animal of bait is dissolved in the water, and aquaculture water is polluted, and causes the objectionable impurities increases such as organism, NH3, H2S in the water body; Chemical oxygen demand (COD) and BOD (BOD) are high simultaneously, can effectively eliminate above-mentioned negative impact through using microbiobacterial agent.Microbial preparation is applied to improve in the aquaculture water ecological setting; Suppress killing microorganism, it also can be used as fodder additives, the extra-nutrition composition; Improve cultivated animals gi tract profitable strain; Reach the purpose of ecological diseases prevention, make the breeding production benign development, obtain better economic benefit and ecological effect.Microbial preparation can effectively reduce nitrite in the cultivation water, ammonia nitrogen, hydrogen sulfide isoconcentration, suppresses harmful microorganism breeding and growth in the water body, purifies water.The metabolism of mikrobe in the preparation own has gasification, ammonification, phosphorus decomposing, denitrification, nitrated and nitrogen fixation; Can pollutent be decomposed into non-toxic substances such as carbonic acid gas, nitrate salt, vitriol; And then be used by the algae in the water body, reach the purpose that purifies water.Its population competition performance suppresses pathogenic bacterium, and useful mattress and host's mucous epithelium are combined closely and generated the compactness mycoderm, form microbial barrier, and the probiotics that has produces microbiotic and bacteriocin kill pathogenic bacteria.
Microbial preparation has little, the high efficiency of investment, advantage such as easy to use.Can full pool spilling head, also can be as fodder additives.Nontoxic, harmless, drug residue free, do not produce resistance, life-time service can reduce antibiotic usage quantity in the breeding process, reduces disease and takes place, and the sewage environmental pollution of discharging is also less.
Summary of the invention
The present invention uses chemical fertilizer, fertilizer in a large number to present aquaculture water; Cause water quality overfertilization, eutrophication serious, bring out that fish raises the nose above water to breathe and problem such as multiple disease, the objective of the invention is to be to provide a kind of complex microorganism preparations that is used for the aquaculture water water quality improvement; Prescription rationally; Easy to use, pollution-free to water body, cost is low.
Another object of the present invention is to be to provide a kind of preparation method who is used for the complex microorganism preparations of aquaculture water water quality improvement, and easy to implement the method, easy and simple to handle, production cost is low.
Technology of the present invention is achieved in that
A kind of complex microorganism preparations that is used for the aquaculture water water quality improvement, it is processed by the raw material of following massfraction:
Described subtilis (seeing the genetic resources table), yeast (seeing the genetic resources table), milk-acid bacteria (seeing the genetic resources table), photosynthetic bacterium (seeing the genetic resources table), nitrobacteria (seeing the genetic resources table), Bacillus licheniformis (seeing the genetic resources table).
A kind of complex microorganism preparations that is used for the aquaculture water water quality improvement, it processes (preferable range) by the raw material of following massfraction:
A kind of complex microorganism preparations that is used for the aquaculture water water quality improvement, it processes (optimum range) by the raw material of following massfraction:
A kind of complex microorganism preparations that is used for the aquaculture water water quality improvement, it processes (best proportioning) by the raw material of following massfraction:
A kind of complex microorganism preparations that is used for the aquaculture water water quality improvement, it processes (best proportioning) by the raw material of following massfraction:
A kind of complex microorganism preparations that is used for the aquaculture water water quality improvement, it processes (best proportioning) by the raw material of following massfraction:
A kind of complex microorganism preparations that is used for the aquaculture water water quality improvement, it processes (best proportioning) by the raw material of following massfraction:
A kind of preparation method who is used for the complex microorganism preparations of aquaculture water water quality improvement the steps include:
A, at first be the cultivation of subtilis, Bacillus licheniformis: subtilis, Bacillus licheniformis are on beef-protein medium; 28-30 ℃; Making primary inclined plane cultivates; Be inoculated into triangular flask then and do vibration secondary liquid culture, be inoculated at last and do three grades on the solid medium and be cultured to the bacterium number and reach 1.0-2.5 * 10
8Individual/the gram solid medium.
B, secondly be saccharomycetic cultivation: yeast is on lactic acid-yam-dextrose culture-medium; 28-30 ℃; Make primary inclined plane and cultivate, be inoculated into then and do vibration secondary liquid culture in the triangular flask, be inoculated at last and do three grades on the solid medium and be cultured to the bacterium number and reach 1.0-2.5 * 10
8Individual/the gram solid medium;
The cultivation of C, milk-acid bacteria: milk-acid bacteria on malt extract medium, 28-30 ℃, make primary inclined plane and cultivate, be inoculated into then and do vibration secondary liquid culture in the triangular flask, be inoculated at last and do three grades on the solid medium and be cultured to the bacterium number and reach 1.0-5.0 * 10
8Individual/the gram solid medium;
The cultivation of D, photosynthetic bacterium: photosynthetic bacterium stoste is inoculated in the liquid test tube with cover, 28-30 ℃, illumination cultivation 3-5 days, gets the photosynthetic bacterium bacteria suspension; Get the bacteria suspension of 1% (volume ratio) and put into the substratum of 500ml triangular flask deactivation; With Fan Shi substratum multiplication culture; 28-30 ℃ of constant-temperature shaking shaking table; Cultivated 36-72 hour; Get the secondary pure growth of photosynthetic bacterium; The photosynthetic bacterium secondary seed of getting 1% (volume ratio) is inoculated in fermentation propagation in the substratum of being made up of 2% (mass ratio) sodium acetate, 1% (mass ratio) ammonium chloride, 0.1% (mass ratio) potassium primary phosphate, 0.5% (mass ratio) sal epsom, 0.2 (mass ratio) sodium-chlor, 3% (mass ratio) amine molybdate, 0.01% (mass ratio) yeast extract, 0.05 (mass ratio) calcium chloride, 0.05% (mass ratio) iron(ic)chloride, 28-30 ℃ of static cultivation of illumination or not illumination shaking culture, liquid photosynthetic bacterium fermenting agent; Again liquid photosynthetic bacterium fermenting agent is inoculated on the solid medium, gets the bacterium number and reach 1.0-2.0 * 10
8The solid photosynthetic bacteria fermenting agent of individual/ml;
The cultivation of E, nitrobacteria: nitrobacteria comprises nitrococcus and nitrifier; Respectively the ammonia-state nitrogen in the water is converted into nitrite and nitrate salt; 24 ℃; Make primary inclined plane and cultivate, be inoculated into then and do vibration secondary liquid culture in the triangular flask, be inoculated at last and do three grades on the solid medium and be cultured to the bacterium number and reach 1.0-2.5 * 10
8Individual/the gram solid medium;
Described nitrococcus substratum: ammonium sulfate
2SO
40.5g, sodium-chlor 0.3g, ferrous sulfate 0.03g, potassium hydrogenphosphate 1g, sal epsom 0.03g, calcium chloride 7.5g, zero(ppm) water 1000ml, PH nature, solid medium add 5% (mass ratio) agar;
Described nitrifier substratum: Sodium Nitrite 1g, sal epsom 0.03g, manganous sulfate 0.01g, potassium hydrogenphosphate 0.75g, yellow soda ash 1g, SODIUM PHOSPHATE, MONOBASIC N0.25g, zero(ppm) water 1000ml, PH nature, solid medium add 5% (mass ratio) agar;
F, subtilis, yeast, milk-acid bacteria, photosynthetic bacterium, nitrobacteria, bacillus licheniformis agent that above-mentioned A, B, C, D, E step are cultivated mix by certain mass ratio; Low temperature (≤60 ℃) drying; Moisture content is controlled at below 20%, just obtains complex microorganism preparations.
The present invention compared with prior art has the following advantages and effect:
Profitable strains such as subtilis, Bacillus licheniformis, photosynthetic bacterium, milk-acid bacteria, yeast, nitrobacteria are carried out scientific and reasonable integration, a kind of complex micro organism fungicide that the uses advanced production technique produces.These cultivate the complex micro organism fungicide form through special methods, have share weal or woe, have complementary functions, the characteristics of cooperation.After applying water body; Utilize the mikrobe metabolic activity, to organic pollutant in the water shift, conversion and Degradation, reduce water pollutant concentration; Objectionable impuritiess such as ammonia nitrogen, nitrous acid, hydrogen sulfide in the collaborative water of decomposition environment are improved the anaerobic environment at the bottom of the pond; Improve the absorb efficient of unicellular algae to nutritive element, promote to swim but the growth of born of the same parents algae, balance algae phase increases the algae variety, suppresses the growth of harmful bacterium.Photosynthetic bacteria utilizes in luminous energy and the water simple inorganics to carry out photosynthesis, fixedly in the water element such as nitrogen and phosphorus the time, produces oxygen oxygen level in the increase water body, raising water transparency; Stablize water pH value.Keep culturing the balance of little ecology, make aquaculture water present " fertilizer ", " work ", " tender ", " feeling well "; Method of use is simple simultaneously, and is convenient, reduces labour cost.
Embodiment
Embodiment 1:
A kind of complex microorganism preparations that is used for the aquaculture water water quality improvement, it is processed by the raw material of following massfraction:
A kind of preparation method who is used for the complex microorganism preparations of aquaculture water water quality improvement the steps include:
A, at first be the cultivation of subtilis, Bacillus licheniformis: subtilis, Bacillus licheniformis are on beef-protein medium; 28 or 29 or 30 ℃; Making primary inclined plane cultivates; Be inoculated into triangular flask then and do vibration secondary liquid culture, be inoculated at last and do three grades on the solid medium and be cultured to the bacterium number and reach 1.0-2.5 * 10
8Individual/the gram solid medium.
B, secondly be saccharomycetic cultivation: yeast is on lactic acid-yam-dextrose culture-medium; 28 or 29 or 30 ℃; Make primary inclined plane and cultivate, be inoculated into then and do vibration secondary liquid culture in the triangular flask, be inoculated at last and do three grades on the solid medium and be cultured to the bacterium number and reach 1.0-2.5 * 10
8Individual/the gram solid medium;
The cultivation of C, milk-acid bacteria: milk-acid bacteria on malt extract medium, 28 or 29 or 30 ℃, make primary inclined plane and cultivate, be inoculated into then and do vibration secondary liquid culture in the triangular flask, be inoculated at last and do three grades on the solid medium and be cultured to the bacterium number and reach 1.0-5.0 * 10
8Individual/the gram solid medium;
The cultivation of D, photosynthetic bacterium: photosynthetic bacterium stoste is inoculated in the liquid test tube with cover, 28-30 ℃, illumination cultivation 3-5 days, gets the photosynthetic bacterium bacteria suspension; Get the bacteria suspension of 1% (volume ratio) and put into the substratum of 500ml triangular flask deactivation; With Fan Shi substratum multiplication culture; 28-30 ℃ of constant-temperature shaking shaking table; Cultivated 36-72 hour; Get the secondary pure growth of photosynthetic bacterium; The photosynthetic bacterium secondary seed of getting 1% (volume ratio) is inoculated in fermentation propagation in the substratum of being made up of 2% (mass ratio) sodium acetate, 1% (mass ratio) ammonium chloride, 0.1% (mass ratio) potassium primary phosphate, 0.5% (mass ratio) sal epsom, 0.2 (mass ratio) sodium-chlor, 3% (mass ratio) amine molybdate, 0.01% (mass ratio) yeast extract, 0.05 (mass ratio) calcium chloride, 0.05% (mass ratio) iron(ic)chloride, 28-30 ℃ of static cultivation of illumination or not illumination shaking culture, liquid photosynthetic bacterium fermenting agent; Again liquid photosynthetic bacterium fermenting agent is inoculated on the solid medium, gets the bacterium number and reach 1.0-2.0 * 10
8The solid photosynthetic bacteria fermenting agent of individual/ml;
The cultivation of E, nitrobacteria: nitrobacteria comprises nitrococcus and nitrifier; Respectively the ammonia-state nitrogen in the water is converted into nitrite and nitrate salt; 24 ℃; Make primary inclined plane and cultivate, be inoculated into then and do vibration secondary liquid culture in the triangular flask, be inoculated at last and do three grades on the solid medium and be cultured to the bacterium number and reach 1.0-2.5 * 10
8Individual/the gram solid medium;
Described nitrococcus substratum: ammonium sulfate
2SO
40.5g, sodium-chlor 0.3g, ferrous sulfate 0.03g, potassium hydrogenphosphate 1g, sal epsom 0.03g, calcium chloride 7.5g, zero(ppm) water 1000ml, PH nature, solid medium add 5% (mass ratio) agar;
Described nitrifier substratum: Sodium Nitrite 1g, sal epsom 0.03g, manganous sulfate 0.01g, potassium hydrogenphosphate 0.75g, yellow soda ash 1g, SODIUM PHOSPHATE, MONOBASIC N0.25g, zero(ppm) water 1000ml, PH nature, solid medium add 5% (mass ratio) agar;
F, subtilis, yeast, milk-acid bacteria, photosynthetic bacterium, nitrobacteria, bacillus licheniformis agent that above-mentioned A, B, C, D, E step are cultivated mix by certain mass ratio; Low temperature (≤60 ℃) drying; Moisture content is controlled at below 20%, just obtains complex microorganism preparations.
Embodiment 2:
A kind of complex microorganism preparations that is used for the aquaculture water water quality improvement, it is processed by the raw material of following massfraction:
Preparation process is identical with embodiment 1.
Embodiment 3:
A kind of complex microorganism preparations that is used for the aquaculture water water quality improvement, it is processed by the raw material of following massfraction:
Preparation process is identical with embodiment 1.
Embodiment 4:
A kind of complex microorganism preparations that is used for the aquaculture water water quality improvement, it is processed by the raw material of following massfraction:
Preparation process is identical with embodiment 1.
Embodiment 5:
A kind of complex microorganism preparations that is used for the aquaculture water water quality improvement, it is processed by the raw material of following massfraction:
Preparation process is identical with embodiment 1.
Embodiment 7:
A kind of complex microorganism preparations that is used for the aquaculture water water quality improvement, it is processed by the raw material of following massfraction:
Preparation process is identical with embodiment 1.
Test case 1:
2 mu of test water surfaces for aquaculture, 1.5 meters of the depth of waters, before using, water body is aging, and fish floating head phenomenon is general, and water transparency has only 15cm.Made the water edge red on June 20th, 2011, mu uses 3KG, with 100 times of clear water dilutions, evenly splashes in the pond, and after 7 days, transparency is brought up to 35cm; Dissolved oxygen rises to 1299mg/L from 772mg/L, and the Cyanophyta living weight drops to 14.832 mg/litre from 24.99 mg/litre; Macrozooplankton reduces relatively, and small-sized wheel animalcule obviously increases, and fish raises the nose above water to breathe phenomenon and disappears basically, water body be improved significantly.
Test case 2:
Test Jiangxia, Wuhan, place, 300 mu of water surfaces for aquaculture, depth of water 1.5-2 rice, before using, the water body algae is main with blue-green algae mainly mutually, blue-green algae quantity 32100 * 10
3Individual/L, water body is aging, and transparency is 20cm, and fish raises the nose above water to breathe in a large number; Made the water edge red on July 12nd, 2011, mu uses 4KG, with 100 times of clear water dilutions, evenly splashes in the pond, and after 7 days, transparency is brought up to 40cm; Detect water quality, blue-green algae quantity reduces to 8600 * 10
3Individual/L, other zooplanktons such as wheel animalcule, cladocera animal are increased, and the water body biological cycle is accelerated, and fish raises the nose above water to breathe phenomenon and disappears basically, water body be improved significantly.
Test case 3:
Test Wuhan, place Hong Shan, 220 mu of water surfaces for aquaculture, depth of water 2-3 rice, before using, body eutrophication is serious, and water body is aging, and transparency is 15cm, and fish raises the nose above water to breathe in a large number; Made the water edge red on July 25th, 2011, mu uses 3KG, with 100 times of clear water dilutions, evenly splashes in the pond, and after 7 days, transparency is brought up to 35cm; The water body biological cycle is accelerated, and fish raises the nose above water to breathe phenomenon and disappears basically, water body be improved significantly.
Claims (10)
1. complex microorganism preparations that is used for the aquaculture water water quality improvement, it is processed by the raw material of following massfraction:
Raw material weight per-cent
Subtilis 5%-50%
Yeast 1%-35%
Milk-acid bacteria 1%-30%
Photosynthetic bacterium 0.5%-30%
Nitrobacteria 0.1%-20%
Bacillus licheniformis 0.5%-25%.
2. a kind of complex microorganism preparations that is used for the aquaculture water water quality improvement according to claim 1, it is characterized in that: it is processed by the raw material of following massfraction:
Raw material weight per-cent
Subtilis 10%-45%
Yeast 5%-28%
Milk-acid bacteria 5%-25%
Photosynthetic bacterium 3%-25%
Nitrobacteria 4%-20%
Bacillus licheniformis 3%-18%.
3. a kind of complex microorganism preparations that is used for the aquaculture water water quality improvement according to claim 1, it is characterized in that: it is processed by the raw material of following massfraction:
Raw material weight per-cent
Subtilis 10%-35%
Yeast 8%-25%
Milk-acid bacteria 10%-20%
Photosynthetic bacterium 8%-20%
Nitrobacteria 10%-20%
Bacillus licheniformis 6%-15%.
4. a kind of complex microorganism preparations that is used for the aquaculture water water quality improvement according to claim 1, it is characterized in that: it is processed by the raw material of following massfraction:
Raw material weight per-cent
Subtilis 10%
Yeast 20%
Milk-acid bacteria 20%
Photosynthetic bacterium 20%
Nitrobacteria 20%
Bacillus licheniformis 10%.
5. a kind of complex microorganism preparations that is used for the aquaculture water water quality improvement according to claim 1, it is characterized in that: it is processed by the raw material of following massfraction:
Raw material weight per-cent
Subtilis 15%
Yeast 15%
Milk-acid bacteria 15%
Photosynthetic bacterium 20%
Nitrobacteria 20%
Bacillus licheniformis 15%.
6. a kind of complex microorganism preparations that is used for the aquaculture water water quality improvement according to claim 1, it is characterized in that: it is processed by the raw material of following massfraction:
Raw material weight per-cent
Subtilis 25%
Yeast 20%
Milk-acid bacteria 10%
Photosynthetic bacterium 20%
Nitrobacteria 10%
Bacillus licheniformis 15%.
7. a kind of complex microorganism preparations that is used for the aquaculture water water quality improvement according to claim 1, it is characterized in that: it is processed by the raw material of following massfraction:
Raw material weight per-cent
Subtilis 30%
Yeast 10%
Milk-acid bacteria 12%
Photosynthetic bacterium 18%
Nitrobacteria 20%
Bacillus licheniformis 10%.
8. the described a kind of preparation method who is used for the complex microorganism preparations of aquaculture water water quality improvement of claim 1 the steps include:
A, at first be the cultivation of subtilis, Bacillus licheniformis: subtilis, Bacillus licheniformis on beef-protein medium, 28-30
0C makes primary inclined plane and cultivates, and is inoculated into triangular flask then and does vibration secondary liquid culture, is inoculated at last to do three grades on the solid medium and be cultured to the bacterium number and reach 1.0-2.5 * 10
8Individual/the gram solid medium;
B, secondly be saccharomycetic cultivation: yeast on lactic acid-yam-dextrose culture-medium, 28-30
0C makes primary inclined plane and cultivates, and is inoculated into then to do vibration secondary liquid culture in the triangular flask, is inoculated at last to do three grades on the solid medium and be cultured to the bacterium number and reach 1.0-2.5 * 10
8Individual/the gram solid medium;
The cultivation of C, milk-acid bacteria: milk-acid bacteria on malt extract medium, 28-30
0C makes primary inclined plane and cultivates, and is inoculated into then to do vibration secondary liquid culture in the triangular flask, is inoculated at last to do three grades on the solid medium and be cultured to the bacterium number and reach 1.0-5.0 * 10
8Individual/the gram solid medium;
The cultivation of D, photosynthetic bacterium: photosynthetic bacterium stoste is inoculated in the liquid test tube with cover 28-30
0C illumination cultivation 3-5 days, gets the photosynthetic bacterium bacteria suspension; Get the bacteria suspension of 1% volume ratio and put into the substratum of 500ml triangular flask deactivation, with Fan Shi substratum multiplication culture, constant-temperature shaking shaking table 28-30
0C; Cultivated 36-72 hour; Get the secondary pure growth of photosynthetic bacterium; The photosynthetic bacterium secondary seed of getting 1% volume ratio is inoculated in fermentation propagation in the substratum of being made up of than iron(ic)chloride than calcium chloride, 0.05% quality than yeast extract, 0.053% quality than amine molybdate, 0.01% quality than sodium-chlor, 3% quality than sal epsom, 0.23% quality than potassium primary phosphate, 0.5% quality than ammonium chloride, 0.1% quality than sodium acetate, 1% quality 2% quality, 28-30
0Static cultivation of C illumination or not illumination shaking culture get liquid photosynthetic bacterium fermenting agent, more liquid photosynthetic bacterium fermenting agent are inoculated on the solid medium, get the bacterium number and reach 1.0-2.0 * 10
8The solid photosynthetic bacteria fermenting agent of individual/ml;
The cultivation of E, nitrobacteria: nitrobacteria comprises nitrococcus and nitrifier; Respectively the ammonia-state nitrogen in the water is converted into nitrite and nitrate salt; 24 ℃; Make primary inclined plane and cultivate, be inoculated into then and do vibration secondary liquid culture in the triangular flask, be inoculated at last and do three grades on the solid medium and be cultured to the bacterium number and reach 1.0-2.5 * 10
8Individual/the gram solid medium;
F, subtilis, yeast, milk-acid bacteria, photosynthetic bacterium, nitrobacteria, bacillus licheniformis agent that above-mentioned A, B, C, D, E step are cultivated mix by mass ratio; Cryodrying; Moisture content is controlled at below 20%, just obtains complex microorganism preparations.
9. a kind of preparation method who is used for the complex microorganism preparations of aquaculture water water quality improvement according to claim 1 is characterized in that: described nitrococcus substratum: ammonium sulfate
2SO
40.5g, sodium-chlor 0.3g, ferrous sulfate 0.03g, potassium hydrogenphosphate 1g, sal epsom 0.03g, calcium chloride 7.5g, zero(ppm) water 1000ml, pH nature, solid medium add 5% quality and compare agar.
10. a kind of preparation method who is used for the complex microorganism preparations of aquaculture water water quality improvement according to claim 1 is characterized in that: described nitrifier substratum: Sodium Nitrite 1g, sal epsom 0.03g, manganous sulfate 0.01g, potassium hydrogenphosphate 0.75g, yellow soda ash 1g, SODIUM PHOSPHATE, MONOBASIC N0.25g, zero(ppm) water 1000ml, pH nature, solid medium add 5% quality and compare agar.
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