CN104719207B - A kind of method of use waste or used plastics bucket culture brachionus plicatilis - Google Patents
A kind of method of use waste or used plastics bucket culture brachionus plicatilis Download PDFInfo
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- CN104719207B CN104719207B CN201510127967.0A CN201510127967A CN104719207B CN 104719207 B CN104719207 B CN 104719207B CN 201510127967 A CN201510127967 A CN 201510127967A CN 104719207 B CN104719207 B CN 104719207B
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Abstract
The invention discloses a kind of method of use waste or used plastics bucket culture brachionus plicatilis, belong to aquaculture field.This technology core is first to be mixed 3 kinds of marine microalgaes with flat round flask, then part cow dung is added in mixing algae solution, rabbit excrement, Saccharomyces cerevisiae powder, minced fish, milk powder, soy sauce grain, wheel animalcule bait liquid is made in chicken manure etc., delivers the air-charging incubation brachionus plicatilis into waste or used plastics bucket of the preprepared equipped with nutrient solution.This technology produces wheel animalcule bait formula science, reasonable, comprehensive nutrition, and wheel animalcule growth is fast, and physique healthy and strong, yield is high；Using waste or used plastics bucket culture wheel animalcule, scrub conveniently, translucency is good, reusable, be difficult pollution, fund is saved in twice laid.
Description
The invention belongs to aquaculture field, more particularly to one kind waste or used plastics bucket culture fold arm tail for technical field
The method of wheel animalcule.
Background technology
Brachionus plicatilis Brachionus plicatilis category wheel animalcules door, single ovary guiding principle, arm tail mesh, Brachionus section,
Brachionusgroup, is a kind of important biological feed.Ocean wheel animalcule has more than 50 kinds, lives in littoral neritic area more.Fold arm tail
Rotifer distribution likes the higher water body of the content of organic matter, to salinity in the brackish water and body of sea water in temperate zone to vast areas of the tropics
Adaptability is very strong, but the tolerance of salinity violent and does not change significantly, optimal salinity 15-25 ‰.17-32 DEG C of thermophilic, water temperature is less than
10 DEG C can produce winter egg, and adult is dead, 25-30 DEG C of optimum temperature.Ingest bacterium, planktonic algae, small-sized protozoan and organic
Chip, 5-25 microns of food particle size is advisable.
Since nineteen sixty finds the living bait that brachionus plicatilis can be used for seawater fish nursery, it is just as fish
Applied in production with crustacean important open-mouthed bait.Report that the wheel animalcule can make 60 kinds of seawater fish and 18 kinds of first at present
The living bait of shell animal nursery.Because the wheel animalcule is nutritious, palatability is strong, reproduction speed is fast, swim slower, suitably do fish,
Shrimp, crab bait, so it is vast aquaculture worker important subject in recent years to carry out brachionus plicatilis batch cultivation.
In wheel animalcule incubation there is provided the sufficient bait of superior in quality, quantity be ensure wheel animalcule grow up healthy and sound, stable high yield it is important
Link, past Chang Fashengyin bait is not enough or cultural method is improper causes that brachionus plicatilis constitution is thin and weak, survival rate is low, a large amount of
The dead or unstable situation of yield.On the cultural method of Brachionus, past Luo Jie etc. discloses a kind of fold arm tail wheel
The cultural method (application number CN201210010461) of worm, the invention brachionus plicatilis cultural method be using discarded or
The old pond of aging is as culture pond, and bait is cultivated wrinkle Brachionus based on small fish, is comprised the concrete steps that
It will cultivate that pond is sterilized, intake after processing of being exposed to the sun, oxygen increasing equipment is installed, then contained foundation bait with mesh bag by a certain amount of
Be hung in culture pond water, oxygen increasing equipment culture 13-18 days combined in daily management, when wheel animalcule density reach 250/
ML, starts to be collected with brachionus plicatilis collecting bag, and breeds speed and density according to brachionus plicatilis and add small fish
Hanging amount.Brachionus plicatilis cultural method of the present invention is simple and convenient, and culture wheel animalcule density is big, in comprehensive nutrition, and water
Fertility is permanent, without frequent bait throwing in, production cost can be greatly lowered, increases economic efficiency.But the method is covered an area of, cultivated
Cycle is long, and practical value is not high.A kind of method (application number of Bdelloid rotifers mass propgation is invented in fourth world etc.
CN200910044826), it is used as bait mass propgation leech shape wheel with the abiotic wheat flour of bait one the invention provides one kind
The method of worm, feature is to use the wheat flour after sterilization treatment to add in a certain amount of sterilized water, wheat flour and sterilized water
W/v is 0.75-0.85g/100mL, i.e., 0.75-0.85 grams of wheat flour is added in every 100 milliliters of sterilized waters, then
Through ultrasonic disruption, then standing sedimentation 24 hours, take supernatant, regulation pH value to 5-7.The bait solution is put into prior preparation
In good wheel animalcule nutrient solution, be placed in 25 DEG C of constant incubators cultivating 12 days, as a result wheel animalcule density up to 100~200/
mL.Feed source of the present invention extensively, it is easy to make, can simplified culture process, reduce cost.But only feed wheat flour, nutrition list
Adjust, wheel animalcule constitution is poor, and yield is not high.In addition, Fan Xinyu etc. discloses a kind of high concentration limnetic chlorella culture ss type folds
The method (application number CN201210120645) of Brachionus, the invention comprises the following steps：Culture vessel is disinfected；Culture
The preparation of base；The wheel animalcule of domestication is inoculated into culture medium, the temperature for keeping culture medium is 28-32 DEG C, and pH is 7-9, dissolved oxygen
DO is more than 8ppm, and salinity is 1.5-2.0%, and persistently supplies high density limnetic chlorella into culture medium, when wheel animalcule density reaches
During to 10000/more than mL, wheel animalcule is collected.The daily growth rate of wheel animalcule reaches 2-10 times in the invention, and density can after being inoculated with 10 days
Up to average 10000-15000/mL, 3,000,000,000 wheel animalcules are turned out with 20 elevated concentrations chlorellas, spawning rate is 30-40%, wheel animalcule
Culture it is very convenient, can obtain purer ultra micro wheel animalcule kind.But the technical matters is complicated, technical requirements are somebody's turn to do, and general workman is difficult
To grasp.
In order to overcome the shortcomings of current technology, the invention provides a kind of use waste or used plastics bucket culture brachionus plicatilis
Method, technological core is that first with 3 kinds of marine microalgaes of flat round flask mixed culture, (eyespot chlorella, lsochrysis zhanjiangensis is micro-
Chlorococcum), part cow dung, rabbit excrement, Saccharomyces cerevisiae powder, minced fish, milk powder, soy sauce grain, chicken manure are then added in mixing algae solution
Etc. wheel animalcule bait liquid is made, the air-charging incubation fold arm tail wheel into waste or used plastics bucket of the preprepared equipped with nutrient solution is delivered
Worm.Compared with conventional art, novelty of the present invention is shown：1) wheel animalcule bait formula science, reasonable, comprehensive nutrition, wheel animalcule life
Long fast, physique healthy and strong, yield is high；2) using waste or used plastics bucket culture wheel animalcule, scrub conveniently, translucency is good, reusable, no
Fund is saved in easily pollution, twice laid；3) wheel animalcule harvests, and the past, speed was slow, and efficiency is low, the present invention frequently with silk cover filtering method
Using the sodium bicarbonate precipitation method, speed is fast, simple to operate, spends few.
The content of the invention
In order to overcome the shortcomings of current technology, the invention provides a kind of use waste or used plastics bucket culture brachionus plicatilis
Method, method is as follows：
The culture culture formula of liquid NaNO of step 1 bait micro-algae30.5 part, KH2PO30.005 part, 1% FeC6H5O7It is molten
0.2 part of liquid, vitamin B10.0002 part, vitamin B120.00002 part, 0.8 part of human urine, 3 parts of ooze Extract, 25- containing ammonia
1 part of 28% ammoniacal liquor, 0.2 part of peptide urea, 0.1 part of ammonium sulfate, 0.5 part of potassium sulfate, 0.8 part of potassium chloride, IBDU 0.3
Part, 0.4 part of oxamides, 0.08 part of ammonium magnesium phosphate, 0.2 part of APP, 0.8 part of sodium metasilicate, 0.0005 part of polyamines, abscisin
0.02 part, 5 parts of sodium acid carbonate, 1.2 parts of soil extract, 0.1 part of medical stone powder boils 1000 parts of Magnetized Seawater after cooling；
One 1500 milliliters of flat round flask is taken, is washed, with distilled water rinse 3 times, above-mentioned 800 milliliters of nutrient solution is added,
With without enemy biological pollution, viability is strong, normal, the eyespot chlorella without obvious sediment that grows vigorous, color, the whip gold such as Zhanjiang
Algae, Nannochloropsis oculata algae kind is inoculated with respectively, is mixed.Inoculum density：50,000 cells/mls of eyespot chlorella, Zhanjiang etc.
40,000 cells/mls of whip chrysophyceae, 80,000 cells/mls of Nannochloropsis oculata；Then clogged and stamped on bottleneck, rubber stopper with rubber stopper
Two holes, insert two thin glass tubes, and 4-6 millimeters of caliber is one longer, for inflating, and insert algae solution, another close to bottom of bottle
Root is shorter, and rubber stopper is exposed at two ends, for being vented, and gas tube one end is connected with air compressor machine, 3-5 liters/min of aeration quantity, protects
Frustule is held to suspend not sink；This round flask is put into microdisk electrode room and cultivated, co-cultures 10 days, cultivated
The 1-3 days of journey, set 18 DEG C of temperature, intensity of illumination 2500lux, light dark period L: D=12h: 12h, i.e., daily 12 small time
According to 12 hours dark.The 4-6 days of incubation, set 21 DEG C of temperature, intensity of illumination 5000lux, light dark period L: D=14h
: 10h, i.e., daily illumination in 14 hours, 10 hours dark.The 7-10 days of incubation, adjust the temperature to 25 DEG C, intensity of illumination
9000lux, light dark period L: D=16h: 8h；By the 7th day, 80 milliliters of fresh medium is added, with the consumption for the salt that supplements the nutrients；
By the 10th day, microalgae density sum can reach 30,000,000 cells/mls, pour out the half of algae solution, and remaining half is used as kind
With addition fresh medium continues to cultivate (i.e. using so-called Semi-continuous cultivation).By the 13rd day, then the half of algae solution is poured out, separately
Half adds fresh medium as kind of a use, continues to cultivate, by the 16th day, then pours out the half of algae solution, second half is used as kind
With, fresh medium is added, continues to cultivate, by the 19th day, stopping culture, all harvesting.
The preparation of step 2 wheel animalcule bait liquid
Algae solution using step 1 harvesting adds part auxiliary element, the bait liquid of wheel animalcule is made as main component.Specifically
It is：1000 parts of algae solution, 2 parts of cow dung, 1 part of rabbit excrement, 0.5 part of Saccharomyces cerevisiae powder, 0.8 part of minced fish, 0.1 part of sesame residues, milk powder 0.3
Part, 0.6 part of soy sauce grain, 0.8 part of chicken manure, 0.3 part of mashed garlic, 0.1 part of honey slag, 0.4 part of Swine blood meal；Stir, use
Mesh aperture is 100 microns of nylon silk cover filtering, waste dregs, the ultrasonication that filtrate is produced with ultrasonic device 3-4 minutes,
Purpose is to be smashed bulky grain bait with ultrasonic wave, becomes little particle, so that wheel animalcule eats, spontaneous fermentation 48 hours, magma
Prepared by the wheel animalcule bait liquid of shape completes, you can for feeding wheel animalcule.
Step 3
The culture of wheel animalcule takes one 25 liters of waste or used plastics bucket (peanut oil drum, Beverage pail or mineral water bottle), and white is thoroughly
It is bright, with sharp printing opacity, soaked 5 hours with 100ppm malachite green solutions, after cleaned down with liquid detergent, test tube, then with distilling
Water rinse 3 times, adds 12 liters and boils cooling Magnetized Seawater, add the wheel animalcule bait liquid of scattered paste shape described in 2 liters of steps 2, shake up, select
Select brachionus plicatilis inoculations active, healthy, without harmful organisms, 25-45/milliliter of inoculum density.Since the 2nd day, often
It adds 0.5 liter of the wheel animalcule bait liquid of scattered paste shape described in step 2.Temperature control is at 23-26 DEG C, illumination 1500-2500lux, from
Bung inserts a thin glass tube inflation, 4-6 millimeters of diameter glass tube.Glass tube is compressed by emulsion tube or plastic tube with air
Machine is connected, and gas componant and ratio are VAir+VPurity oxygen=99%+1% (volume ratio).4-8 liters/min of aeration quantity, aeration quantity is too big
It is unfavorable for ingesting and living for wheel animalcule on the contrary.By the culture disposed within of this Plastic Drum, air-conditioning temperature control, CFL lighting.Cultivate
12nd day, wheel animalcule density was up to 2000-5000/milliliter, you can harvesting.
The harvesting of step 4 wheel animalcule
Wheel animalcule nutrient solution described in step 3 is taken with 10 liters of graduated cylinder, adds sodium bicarbonate (NaHCO by every liter of nutrient solution3) 3.5 grams
Amount adds sodium bicarbonate, stirs, staticly settles 24 hours, rapid rise and the heat of release due to medium pH value, wheel
Worm shock, stupor, sink to bottom, abandon supernatant, obtain the wheel animalcule of bottom deposit, then with 10% acetic acid solution (volume ratio)
Neutralize, adjust pH value to 7.0-8.2, shake up, reprecipitation 24 hours abandons supernatant, obtains wheel animalcule, you can to feed scallop, fish
The economic animals such as seedling, prawn, swimming crab.
Beneficial outcomes
In order to overcome the shortcomings of current technology, the invention provides a kind of use waste or used plastics bucket culture brachionus plicatilis
Method, technological core is that first with 3 kinds of marine microalgaes of flat round flask mixed culture, (eyespot chlorella, lsochrysis zhanjiangensis is micro-
Chlorococcum), part cow dung, rabbit excrement, Saccharomyces cerevisiae powder, minced fish, milk powder, soy sauce grain, chicken manure are then added in mixing algae solution
Etc. wheel animalcule bait liquid is made, the air-charging incubation fold arm tail wheel into waste or used plastics bucket of the preprepared equipped with nutrient solution is delivered
Worm.Compared with conventional art, novelty of the present invention is shown：1) wheel animalcule bait formula science, reasonable, comprehensive nutrition, wheel animalcule life
Long fast, physique healthy and strong, yield is high；2) using waste or used plastics bucket culture wheel animalcule, scrub conveniently, translucency is good, reusable, no
Fund is saved in easily pollution, twice laid；3) wheel animalcule harvests, and the past, speed was slow, and efficiency is low, the present invention frequently with silk cover filtering method
Using the sodium bicarbonate precipitation method, speed is fast, simple to operate, spends few.
Brief description of the drawings
Fig. 1 brachionus plicatilis aspect graphs；
The method of Fig. 2 round flask culture bait micro-algaes；Arrow meaning is charge air conditioning trend；
Fig. 3 waste or used plastics bucket culture brachionus plicatilis methods；Arrow meaning is charge air conditioning trend.
Embodiment
A kind of method of use waste or used plastics bucket culture brachionus plicatilis, step is as follows：
The culture of step 1 bait micro-algae
Culture formula of liquid is NaNO by weight30.5 part, KH2PO30.005 part, 1% FeC6H5O7Solution 0.2
Part, vitamin B10.0002 part, vitamin B120.00002 part, 0.8 part of human urine, 3 parts of ooze Extract, the 25-28%'s containing ammonia
1 part of ammoniacal liquor, 0.2 part of peptide urea, 0.1 part of ammonium sulfate, 0.5 part of potassium sulfate, 0.8 part of potassium chloride, 0.3 part of IBDU, grass
0.4 part of acid amides, 0.08 part of ammonium magnesium phosphate, 0.2 part of APP, 0.8 part of sodium metasilicate, 0.0005 part of polyamines, 0.02 part of abscisin,
5 parts of sodium acid carbonate, 1.2 parts of soil extract, 0.1 part of medical stone powder boils 1000 parts of Magnetized Seawater after cooling；Take one
1500 milliliters of flat round flask, is washed, with distilled water rinse 3 times, adds above-mentioned 800 milliliters of nutrient solution, with unmatched
Evil biological pollution, viability are by force, growth is vigorous, color is normal, the eyespot chlorella without obvious sediment, and lsochrysis zhanjiangensis is micro-
Chlorococcum algae kind is inoculated with respectively, is mixed；Inoculum density：50,000 cells/mls of eyespot chlorella, the whip such as Zhanjiang gold
40,000 cells/mls of algae, 80,000 cells/mls of Nannochloropsis oculata；Then clogged with rubber stopper and two are stamped on bottleneck, rubber stopper
Hole, inserts two thin glass tubes, 4-6 millimeter of caliber is one longer, for inflating, insertion algae solution, close to bottom of bottle, another compared with
Short, rubber stopper is exposed at two ends, for being vented, and gas tube one end is connected with air compressor machine, 3-5 liters/min of aeration quantity, keeps algae
Cell suspends not sink；This round flask is put into microdisk electrode room and cultivated, is co-cultured 10 days, incubation
The 1-3 days, 18 DEG C of temperature, intensity of illumination 2500lux, light dark period L: D=12h: 12h, i.e., daily illumination in 12 hours, 12 were set
Hour is dark；The 4-6 days of incubation, 21 DEG C of temperature of setting, intensity of illumination 5000lux, light dark period L: D=14h: 10h,
I.e. daily illumination in 14 hours, 10 hours dark；The 7-10 days of incubation, adjust the temperature to 25 DEG C, intensity of illumination
9000lux, light dark period L: D=16h: 8h；By the 7th day, 80 milliliters of fresh medium is added, with the consumption for the salt that supplements the nutrients；
By the 10th day, microalgae density sum can reach 30,000,000 cells/mls, pour out the half of algae solution, and remaining half is used as kind
With addition fresh medium continues to cultivate (i.e. using so-called Semi-continuous cultivation)；By the 13rd day, then the half of algae solution is poured out, separately
Half adds fresh medium as kind of a use, continues to cultivate, by the 16th day, then pours out the half of algae solution, second half is used as kind
With, fresh medium is added, continues to cultivate, by the 19th day, stopping culture, all harvesting；
The preparation of step 2 wheel animalcule bait liquid
Algae solution using step 1 harvesting adds part auxiliary element, the bait liquid of wheel animalcule is made as main component；Specifically
It is：1000 parts of algae solution, 2 parts of cow dung, 1 part of rabbit excrement, 0.5 part of Saccharomyces cerevisiae powder, 0.8 part of minced fish, 0.1 part of sesame residues, milk powder 0.3
Part, 0.6 part of soy sauce grain, 0.8 part of chicken manure, 0.3 part of mashed garlic, 0.1 part of honey slag, 0.4 part of Swine blood meal；Stir, use
Mesh aperture is 100 microns of nylon silk cover filtering, waste dregs, the ultrasonication that filtrate is produced with ultrasonic device 3-4 minutes,
Purpose is to be smashed bulky grain bait with ultrasonic wave, becomes little particle, so that wheel animalcule eats, spontaneous fermentation 48 hours, magma
Prepared by the wheel animalcule bait liquid of shape completes, you can for feeding wheel animalcule；
The culture of step 3 wheel animalcule
Take one 25 liters of waste or used plastics bucket (peanut oil drum, Beverage pail or mineral water bottle), white clear, with it is sharp thoroughly
Light, is soaked 5 hours with 100ppm malachite green solutions, after cleaned down with liquid detergent, test tube, then with distilled water rinse 3 times,
Add 12 liters and boil cooling Magnetized Seawater, add the wheel animalcule bait liquid of scattered paste shape described in 2 liters of steps 2, shake up, select active, strong
Health, the brachionus plicatilis inoculation without harmful organisms, 25-45/milliliter of inoculum density.Since the 2nd day, step was added daily
0.5 liter of the wheel animalcule bait liquid of scattered paste shape described in 2；Temperature control is in 23-26 DEG C, illumination 1500-2500lux, from bung insertion one
Root thin glass tube is inflated, 4-6 millimeters of diameter glass tube；Glass tube is connected by emulsion tube or plastic tube with air compressor, gas
Body composition and ratio are VAir+VPurity oxygen=99%+1% (volume ratio).4-8 liters/min of aeration quantity, aeration quantity is too big unfavorable on the contrary
Ingesting and living in wheel animalcule；By the culture disposed within of this Plastic Drum, air-conditioning temperature control, CFL lighting.Cultivate by the 12nd day, take turns
Worm density is up to 2000-5000/milliliter, you can harvesting；
The harvesting of step 4 wheel animalcule
Wheel animalcule nutrient solution described in step 3 is taken with 10 liters of graduated cylinder, adds sodium bicarbonate (NaHCO by every liter of nutrient solution3) 3.5 grams
Amount adds sodium bicarbonate, stirs, staticly settles 24 hours, rapid rise and the heat of release due to medium pH value, wheel
Worm shock, stupor, sink to bottom, abandon supernatant, obtain the wheel animalcule of bottom deposit, then with 10% acetic acid solution (volume ratio)
Neutralize, adjust pH value to 7.0-8.2, shake up, reprecipitation 24 hours abandons supernatant, obtains wheel animalcule, you can to feed scallop, fish
Seedling, prawn, swimming crab economic animal.
FeC described in step 16H5O7(ironic citrate) compared with indissoluble solution, can add a small amount of running water on stove low-grade fever to 80-
90 DEG C, and be stirred continuously to whole thawings；
Medical stone powder described in step 1 crushes gained by the size-reduced machine of medical stone, and particle diameter is less than 25 microns；
Human urine preparation method described in step 1：1000 milliliters of human urine is taken, conical flask was boiled after 5 minutes, cooled down standby；
Soil extract preparation method described in step 1：The woods 1000 parts of soil at the middle and upper levels, plus 1000 parts of distilled water are taken,
Again plus 2.5 parts of NaOH, heating, boils 45 minutes, cooling, precipitation 12 hours, abandons sediment, takes supernatant, filter paper filtering, filtrate
As soil extract；
Ooze described in step 1 extracts liquid and preparation method thereof：Take comparison fertile and the not black ooze of what, with 1 part of mud add 2 parts it is light
Water, is stirred, and is stood 1-2 minutes, is taken upper strata mud in aluminum pot, 1 gram of NaOH amount is added by every 1000 milliliters of mud
NaOH is added, is boiled 25 minutes.It need to be stirred continuously uniformly, be cooled down after boiling, standing takes supernatant standby in 24 hours when boiling；
NaNO described in step 13, KH2PO3, FeC6H5O7, peptide urea, ammonium sulfate, potassium sulfate, the chemical drugs such as potassium chloride
Product purity is both needed to chemical pure rank；
Magnetized water preparation method used in microalgae is cultivated in step 1：After seawater boils, cooling is flowed through after water magnetizing apptss, is
Magnetized water.Water magnetizing apptss magnetic induction intensity is 0.3T, and flow is 3t/h, and flow velocity is 0.2m/s.Magnetic used in wheel animalcule is cultivated in step 3
Change water to be made with same method；
Cow dung preparation method described in step 2：50 kilograms of cow dung is taken, 3 kilograms of wheat bran, 1 kilogram of maize flour, running water is added
4 liters, it is sufficiently stirred for, piles coniform, spontaneous fermentation 15 days, spread out be exposed to the sun 4-5 days under straticulation, sunlight afterwards, pulverizer is crushed, grain
Footpath is less than 25 microns；
Ultrasonic treatment time 4-5 described in step 2 minute, dutycycle 0.7, ultrasound intensity 250W/cm2；
Rabbit excrement described in step 2 and chicken manure need to be respectively charged into polybag spontaneous fermentation 7-9 days, then be poured on concrete floor,
Spread out straticulation, be exposed to the sun 3-5 days under sunlight, after powder is broken into pulverizer, particle diameter is less than 25 microns.
Finally it should be noted that：The above embodiments are merely illustrative of the technical solutions of the present invention, rather than its limitations；Although
The present invention is described in detail with reference to the foregoing embodiments, it will be understood by those within the art that：It still may be used
To be modified to the technical scheme described in previous embodiment, or equivalent substitution is carried out to which part technical characteristic；And
These modifications are replaced, and the essence of appropriate technical solution is departed from the spirit and model of technical scheme of the embodiment of the present invention
Enclose.
Claims (1)
1. a kind of method of use waste or used plastics bucket culture brachionus plicatilis, it is characterised in that step is as follows：
The culture of step 1 bait micro-algae
Culture formula of liquid is NaNO by weight30.5 part, KH2PO30.005 part, 1% FeC6H5O70.2 part of solution, dimension life
Plain B10.0002 part, vitamin B120.00002 part, 0.8 part of human urine, 3 parts of ooze Extract, 1 part of the ammoniacal liquor of the 25-28% containing ammonia,
0.2 part of peptide urea, 0.1 part of ammonium sulfate, 0.5 part of potassium sulfate, 0.8 part of potassium chloride, 0.3 part of IBDU, oxamides 0.4
Part, 0.08 part of ammonium magnesium phosphate, 0.2 part of APP, 0.8 part of sodium metasilicate, 0.0005 part of polyamines, 0.02 part of abscisin, bicarbonate
5 parts of sodium, 1.2 parts of soil extract, 0.1 part of medical stone powder boils 1000 parts of Magnetized Seawater after cooling；Take one 1500 milliliters
Flat round flask, wash, with distilled water rinse 3 times, add above-mentioned 800 milliliters of nutrient solution, with dirty without harmful organisms
Dye, viability are strong, growth is vigorous, color is normal, the eyespot chlorella without obvious sediment, lsochrysis zhanjiangensis, Nannochloropsis oculata algae
Plant and be inoculated with respectively, be mixed；Inoculum density：50,000 cells/mls of eyespot chlorella, lsochrysis zhanjiangensis is 40,000 thin
Born of the same parents/milliliter, 80,000 cells/mls of Nannochloropsis oculata；Then clogged with rubber stopper and two holes are stamped on bottleneck, rubber stopper, insertion two
Root thin glass tube, 4-6 millimeters of caliber is one longer, for inflating, and inserts algae solution, another shorter close to bottom of bottle, two ends dew
Go out rubber stopper, for being vented, gas tube one end is connected with air compressor machine, 3-5 liters/min of aeration quantity, keep frustule to suspend
Do not sink；This round flask is put into microdisk electrode room and cultivated, co-cultivation 10 days, the 1-3 days of incubation,
18 DEG C of temperature of setting, intensity of illumination 2500lux, light dark period L: D=12h: 12h, i.e., daily illumination in 12 hours, 12 hours are black
Secretly；The 4-6 days of incubation, set 21 DEG C of temperature, and intensity of illumination 5000lux, light dark period L: D=14h: 10h is that is, daily
Illumination in 14 hours, 10 hours dark；The 7-10 days of incubation, adjust the temperature to 25 DEG C, intensity of illumination 9000lux, brightness
Period L: D=16h: 8h；By the 7th day, 80 milliliters of fresh medium is added, with the consumption for the salt that supplements the nutrients；To the 10th day, microalgae
Density sum can reach 30,000,000 cells/mls, pour out the half of algae solution, and remaining half adds fresh training as kind of a use
Nutrient solution, continues to cultivate, i.e., using so-called Semi-continuous cultivation；By the 13rd day, then pour out the half of algae solution, second half as kind of a use,
Fresh medium is added, continues to cultivate, by the 16th day, then the half of algae solution is poured out, second half adds fresh cultured as kind of a use
Liquid, continued to cultivate, by the 19th day, stopped culture, all harvesting；
The preparation of step 2 wheel animalcule bait liquid
Algae solution using step 1 harvesting adds part auxiliary element, the bait liquid of wheel animalcule is made as main component；
Specifically：1000 parts of algae solution, 2 parts of cow dung, 1 part of rabbit excrement, 0.5 part of Saccharomyces cerevisiae powder, 0.8 part of minced fish, sesame residues 0.1
Part, 0.3 part of milk powder, 0.6 part of soy sauce grain, 0.8 part of chicken manure, 0.3 part of mashed garlic, 0.1 part of honey slag, 0.4 part of Swine blood meal；Fully stir
Mix uniform, with the nylon silk cover filtering that mesh aperture is 100 microns, waste dregs, at the ultrasonic wave that filtrate is produced with ultrasonic device
Reason 3-4 minutes, it is therefore an objective to bulky grain bait is smashed with ultrasonic wave, becomes little particle, so that wheel animalcule eats, spontaneous fermentation 48
Hour, prepared by the wheel animalcule bait liquid of scattered paste shape completes, you can for feeding wheel animalcule；
The culture of step 3 wheel animalcule
One 25 liters of waste or used plastics bucket is taken, white clear with sharp printing opacity, is soaked 5 hours with 100ppm malachite green solutions, rear to use
Liquid detergent, test tube are cleaned down, then with distilled water rinse 3 times, are added 12 liters and boiled cooling Magnetized Seawater, add 2 liters of steps 2
The wheel animalcule bait liquid of the scattered paste shape, shakes up, and selection is active, health, the brachionus plicatilis without harmful organisms are inoculated with, and is inoculated with close
Spend 25-45/milliliter；Since the 2nd day, 0.5 liter of the wheel animalcule bait liquid of scattered paste shape described in step 2 was added daily；Temperature control
At 23-26 DEG C, illumination 1500-2500lux inserts a thin glass tube inflation, 4-6 millimeters of diameter glass tube from bung；Glass
Pipe is connected by emulsion tube or plastic tube with air compressor, and gas componant and ratio are VAir+VPurity oxygen=99%+1% (volumes
Than)；4-8 liters/min of aeration quantity, aeration quantity is unfavorable for ingesting and living for wheel animalcule very much on the contrary greatly；This Plastic Drum is disposed within
Culture, air-conditioning temperature control, CFL lighting；Cultivate by the 12nd day, wheel animalcule density is up to 2000-5000/milliliter, you can harvesting；
The harvesting of step 4 wheel animalcule
Wheel animalcule nutrient solution described in step 3 is taken with 10 liters of graduated cylinder, adds sodium bicarbonate NaHCO by every liter of nutrient solution33.5 grams of amount is added
Sodium bicarbonate, stirs, and staticly settles 24 hours, rapid rise and the heat of release due to medium pH value, wheel animalcule shock,
Stupor, sinks to bottom, abandons supernatant, obtain the wheel animalcule of bottom deposit, is then neutralized, adjusted with 10% acetic acid solution (volume ratio)
Save pH value to 7.0-8.2, shake up, reprecipitation 24 hours abandons supernatant, obtains wheel animalcule, you can to feed scallop, fry, prawn,
Swimming crab economic animal；
FeC described in step 16H5O7(ironic citrate) compared with indissoluble solution, plus a small amount of running water on stove low-grade fever to 80-90 DEG C, and
It is stirred continuously to whole thawings；
Medical stone powder described in step 1 crushes gained by the size-reduced machine of medical stone, and particle diameter is less than 25 microns；
Human urine preparation method described in step 1：1000 milliliters of human urine is taken, conical flask was boiled after 5 minutes, cooled down standby；
Soil extract preparation method described in step 1：The woods 1000 parts of soil at the middle and upper levels, plus 1000 parts of distilled water are taken, then is added
2.5 parts of NaOH, heating, boils 45 minutes, cooling, precipitation 12 hours, abandons sediment, takes supernatant, filter paper filtering, and filtrate is
Soil extract；
Ooze described in step 1 extracts liquid and preparation method thereof：Take comparison fertile and the not black ooze of what, 2 parts of fresh water added with 1 part of mud,
Stir, stand 1-2 minutes, take upper strata mud in aluminum pot, the amount for adding 1 gram of NaOH by every 1000 milliliters of mud is added
NaOH, boils 25 minutes；It need to be stirred continuously uniformly, be cooled down after boiling, standing takes supernatant standby in 24 hours when boiling；
NaNO described in step 13, KH2PO3, FeC6H5O7, peptide urea, ammonium sulfate, potassium sulfate, potassium chloride purity is both needed to chemistry
Pure rank；
Magnetized water preparation method used in microalgae is cultivated in step 1：After seawater boils, cooling is flowed through after water magnetizing apptss, as magnetized
Water；Water magnetizing apptss magnetic induction intensity is 0.3T, and flow is 3t/h, and flow velocity is 0.2m/s；Magnetized water used in wheel animalcule is cultivated in step 3
It is made with same method；
Cow dung preparation method described in step 2：Take 50 kilograms of cow dung, add 3 kilograms of wheat bran, 1 kilogram of maize flour, 4 liters of running water,
It is sufficiently stirred for, piles coniform, spontaneous fermentation 15 days, spread out be exposed to the sun 4-5 days under straticulation, sunlight afterwards, pulverizer is crushed, particle diameter is small
In 25 microns；
Ultrasonic treatment time 4-5 described in step 2 minute, dutycycle 0.7, ultrasound intensity 250W/cm2；
Rabbit excrement described in step 2 and chicken manure need to be respectively charged into polybag spontaneous fermentation 7-9 days, are then poured on concrete floor, spread out into
Be exposed to the sun under thin layer, sunlight 3-5 days, after break into powder with pulverizer, particle diameter is less than 25 microns.
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| CN109169434A (en) * | 2018-09-18 | 2019-01-11 | 广西壮族自治区水产科学研究院 | A kind of freshwater wheel animalcule fast breeding method |
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