CN1962488A - Composite microecological agent for aquaculture and scenery water body and its preparation method - Google Patents
Composite microecological agent for aquaculture and scenery water body and its preparation method Download PDFInfo
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- CN1962488A CN1962488A CN 200610098257 CN200610098257A CN1962488A CN 1962488 A CN1962488 A CN 1962488A CN 200610098257 CN200610098257 CN 200610098257 CN 200610098257 A CN200610098257 A CN 200610098257A CN 1962488 A CN1962488 A CN 1962488A
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Abstract
The invention discloses a compound micro-ecological agent and preparing method of aquaculture and landscape water body, which is allocated by aquatic candida, ball-shaped yeast, bacillus subtilis and shipseudomonas with quality rate at 0.5-1.7:0.0-1.5:1.0-3.0:1.0-2.5. The agent can decompose eutrophication material in the aquaculture and landscape water body, which inhibits blue-green algae effectively.
Description
Technical field
The present invention relates to a kind of aquaculture and scenery water body compound micro-ecological preparation, belong to aquatic products microbiology and ecological goods and manufacturing field thereof.
Background technology
The general mode that adopts high-density and intensification in the breeding process of fishery products, make many objectionable impuritiess accumulation in the water body, thereby endanger the aquaculture organism in the water body directly or indirectly, little Ecological Control technology is because numerous advantages obtain people's attention day by day, and domestic have a lot of products to sell.But there are many limitation in currently available products: the scope of microbial strains adaptive temperature is narrower; Thalline is too little, and the flocculation proterties is not so good, not easily collecting; Bacterial classification quantity competitive list one, not strong to the adaptability of various water quality types, range of application is wide inadequately; Problems such as do not consider its safety issue to aquaculture organisms, there is the capability and performance instability in the product of extensive style solid state fermentation and liquid fermenting production, and living bacteria count is few.
Summary of the invention
What the present invention will solve is exactly the problem that above-mentioned prior art exists, and a kind of aquaculture and scenery water body compound micro-ecological preparation are provided.
The present invention presses 0.5-1.7 by aquatic candiyeast (Candida aquatica), Saccharomyces globosus (Saccharomyces globosus), subtilis (Bacillus subtilis) and these four kinds of bacterium of Pseudomonas stutzeri (Pseudomonas stutzeri) and substratum separately: the mass ratio of 0.0-1.5: 1.0-3.0: 1.0-2.5 is formulated.
The seed of each bacterial classification and fermention medium be by the formulated aqueous solution of following material, the nutrient media components scope following (unit: g/L):
Aquatic candiyeast: molasses 5-15, peptone 0-1, glucose 5-15, ammonia chloride 1-4, yeast extract paste 0.1-0.5, potassium primary phosphate 0.2-1.0, dipotassium hydrogen phosphate 0.5-1.5, sal epsom 0.3-0.8, sodium-chlor 0.4-0.7, ferrous sulfate (0.001-0.005) * 10
-3, water 1L.
Saccharomyces globosus: molasses 6-14, peptone 0-2, glucose 7-16, ammonia chloride 1.5-3.0, yeast extract paste 0.2-0.7, potassium primary phosphate 0.3-0.8, dipotassium hydrogen phosphate 0.4-1.5, sal epsom 0.35-0.7, sodium-chlor 0.3-0.9, ferrous sulfate (0.002-0.005) * 10
-3, water 1L.
Subtilis: molasses 2-13, peptone 1-6, glucose 2-8, ammonia chloride 0.5-3.0, yeast extract paste 0.15-0.45, potassium primary phosphate 0.25-0.6, dipotassium hydrogen phosphate 0.6-0.9, sal epsom 0.2-1.0, sodium-chlor 0.2-1.3, ferrous sulfate (0.001-0.004) * 10
-3, water 1L.
Pseudomonas stutzeri: molasses 6-17, peptone 1.5-4, glucose 3-10, ammonia chloride 0.7-2.5, yeast extract paste 0.21-0.7, potassium primary phosphate 0.1-0.7, dipotassium hydrogen phosphate 0.3-1.2, sal epsom 0.4-0.9, sodium-chlor 0.1-0.8, ferrous sulfate (0.002-0.007) * 10
-3, water 1L.
Preparation method of the present invention comprises the steps:
1) shakes a bottle rejuvenation for a short time
Use transfering loop respectively under aseptic technique on the inclined-plane of 4 kinds of bacterium, be inoculated in and contain the shaking for a short time in the bottle of corresponding substratum, in shaking table, carry out pure culture respectively;
2) shake the bottle amplification greatly
Under aseptic technique, be transferred to respectively and contain the shaking greatly in the bottle of corresponding substratum shaking 4 kinds of qualified bacterium of cultivation in the bottle for a short time, in shaking table, carry out pure culture respectively;
3) preparation of seeding tank fermention medium
Preparation seeding tank fermention medium, the steam high-temperature sterilization postcooling that carries out seeding tank and associated conduit is to temperature required;
4) seeding tank fermentation
Insert respectively through check to 4 seeding tanks by the inoculation valve rapidly under the flame protection and cultivate qualified seed liquor, inoculum size is 10%-20%;
5) second order fermentation jar medium preparation
Preparation second order fermentation jar fermention medium is also put into, and fermentor tank and associated conduit are carried out steam high-temperature sterilization postcooling to temperature required;
6) second order fermentation jar large scale fermentation
By pressure that reduces the second order fermentation jar and the pressure that increases seeding tank, the seed liquor of 4 seeding tanks that will be through being up to the standards moves in the second order fermentation jar carries out multiplication culture, and the culture transferring amount is 10%-20%
7) finished product can
The bacterium liquid of second order fermentation jar that will be through being up to the standards carries out can after moving in proportion and carrying out aeration-agitation mixing 5-12h in the sterilized storage tank then between sterile filling.
Abovementioned steps 1) and 2) culture condition be temperature 27-37 ℃, rotating speed 110-220r/min, incubation time are 17-24h; Requirement microscopy bacterium is pollution-free and form is normal, and every kind of bacterium viable count is not less than 10
8/ mL.
Abovementioned steps 1) and step 2) in the consumption of substratum be respectively 30mL and 500mL, the capacity that shakes bottle for a short time in the step 1) is 150mL, step 2) in the capacity that shakes bottle greatly be 2500mL.
Abovementioned steps 3) temperature condition of steam high-temperature sterilization is and in the step 5), and 121 ℃ of conditions continue to stop behind the 20-30min, to the fermentor tank pressurize between the 0.02-0.18MPa and be cooled between 27-37 ℃.
Aforementioned fermentation condition in step 4) is: temperature 27-37 ℃, stirrer rotating speed 110-220r/min, incubation time are 17-24h,
Air flow 4m
3/ h, tank pressure 0.02-0.18MPa, the pH value is 3.0-7.0; Requirement microscopy bacterium is pollution-free and form is normal, and every kind of bacterium viable count is not less than 10
8/ mL.
Abovementioned steps 6) fermentation condition in: temperature 27-37 ℃, stirrer rotating speed 110-220r/min, incubation time are 17-24h, air flow 4m
3/ h, the tank pressure hold-in range is 0.02-0.18MPa.Requirement microscopy bacterium is pollution-free and form is normal, and every kind of bacterium viable count is not less than 10
8/ mL.
Every kind of bacterium viable count is not less than 10 in the finished product that step 7) makes
8/ mL.
Because 4 kinds of contained bacterium of the present invention are viable bacteria, therefore it also can secrete some other useful factors of generation, as enzyme, amino acid, vitamin H, VITAMIN etc., these the 4 kinds of bacterium and the useful factor of excretory thereof can be decomposed the deleterious material of cultivated animals, as the nutriment of planktonic organism and cultivated animals, promote its growth, can improve the immunizing power of cultivated animals, preventing disease etc.Therefore, the present invention can degrade and transform the eutrophication material of aquaculture and landscape water body, purify water, and have the function that suppresses the blue-green algae in the water body, can be used for various seas, freshwater aquiculture (shrimp, crab, fish, shellfish) animal nursery pond and growing pond and some eutrophication waters etc.The present invention has overcome many shortcomings of currently available products, it is wide to have the adaptive temperature scope, bacterial classification quantity is abundant, types of spawn is many, and stronger to the adaptability of various water quality types, range of application is wide, considered safety issue to aquaculture organisms, and the sterile production method of employing enclosed liquid deep layer fermenting process and whole process etc. has guaranteed the quality and the performance of product, has improved the living bacteria count height greatly, and is actual in 10
8/ mL is with first-class.
Embodiment
Embodiment 1: the preparation of compound micro-ecological preparation fermention medium
The fermention medium of 4 bacterial classifications according to following requirement prepare (unit: g/L):
Aquatic candiyeast: molasses 10, glucose 10, ammonia chloride 1, yeast extract paste 0.2, potassium primary phosphate 0.6, dipotassium hydrogen phosphate 0.7, sal epsom 0.4, sodium-chlor 0.5, ferrous sulfate 4 * 10
-3, water 1L.
Saccharomyces globosus: molasses 10, glucose 10, ammonia chloride 1.5, yeast extract paste 0.3, potassium primary phosphate 0.5, dipotassium hydrogen phosphate 0.7, sal epsom 0.5, sodium-chlor 0.5, ferrous sulfate 5 * 10
-3, water 1L.
Subtilis: molasses 10, peptone 2, glucose 5, ammonia chloride 1, yeast extract paste 0.35, potassium primary phosphate 0.4, dipotassium hydrogen phosphate 0.7, sal epsom 0.5, sodium-chlor 0.5, ferrous sulfate 4.5 * 10
-3, water 1L.
Pseudomonas stutzeri: molasses 10, peptone 2, glucose 5, ammonia chloride 0.8, yeast extract paste 0.4, potassium primary phosphate 0.3, dipotassium hydrogen phosphate 0.5, sal epsom 0.5, sodium-chlor 0.5, ferrous sulfate 5.5 * 10
-3, water 1L.
Raw material according to aforementioned proportion difference weighing 100L mixes the back and carry out high-temperature sterilization in fermentor tank, is cooled to be fit to temperature,
Inoculum size 10%, pH7.0, air flow 4m
3/ h, 30 ℃ of culture temperature are fermented in 4 100L fermentor tanks respectively under the condition of incubation time 24h, during fermentation ends, by plate dilution method detect subtilis, Pseudomonas stutzeri, aquatic candiyeast and
The biomass of Saccharomyces globosus is respectively 1 * 10
9Individual/mL, 3 * 10
9Individual/mL, 9 * 10
8Individual/mL and 2 * 10
8Individual/mL.
Embodiment 2: the preparation of compound micro-ecological preparation seed culture medium
The big shake-flask seed substratum of 4 bacterial classifications according to following requirement prepare (unit: g/L):
Aquatic candiyeast: molasses 10, glucose 10, ammonia chloride 1, yeast extract paste 0.2, potassium primary phosphate 0.6, dipotassium hydrogen phosphate 0.7, sal epsom 0.4, sodium-chlor 0.5, ferrous sulfate 4 * 10
-3, water 1L.
Saccharomyces globosus: molasses 10, glucose 10, ammonia chloride 1.5, yeast extract paste 0.3, potassium primary phosphate 0.5, dipotassium hydrogen phosphate 0.7, sal epsom 0.5, sodium-chlor 0.5, ferrous sulfate 5 * 10
-3, water 1L.
Subtilis: molasses 10, peptone 2, glucose 5, ammonia chloride 1, yeast extract paste 0.35, potassium primary phosphate 0.4, dipotassium hydrogen phosphate 0.7, sal epsom 0.5, sodium-chlor 0.5, ferrous sulfate 4.5 * 10
-3, water 1L.
Pseudomonas stutzeri: molasses 10, peptone 2, glucose 5, ammonia chloride 0.8, yeast extract paste 0.4, potassium primary phosphate 0.3, dipotassium hydrogen phosphate 0.5, sal epsom 0.5, sodium-chlor 0.5, ferrous sulfate 5.5 * 10
-3, water 1L.
Raw material according to aforementioned proportion difference weighing 500mL, mix the back and carry out high-temperature sterilization in the bottle shaking greatly, be cooled to be fit to temperature, with shake for a short time in the bottle through microscopy cultivate 4 kinds of qualified bacterium respectively under aseptic technique the ratio in inoculum size 15% be transferred to shaking greatly in the bottle of 2500mL, in shaking table, cultivate, condition is 30 ℃ of culture temperature, air flow is 100%, incubation time is 24h, shaking speed 170r/min, cultivate when finishing, detect subtilis by plate dilution method, Pseudomonas stutzeri, the biomass of aquatic candiyeast and Saccharomyces globosus is respectively 2 * 10
9Individual/mL, 3 * 10
9Individual/mL, 5 * 10
8Individual/mL and 3 * 10
8Individual/mL.
Embodiment 3: compound micro-ecological preparation is produced fermentation parameter research
Under different inoculum sizes, air flow, initial pH value, culture temperature and incubation time, the different microorganism of 4 strains is carried out cultivation and fermentation, tunning is carried out enumeration, with the screening best fermentation condition.6h after fermentation, 9h, 12h, 15h, 18h and 24h analyze the variation of the output of the dynamic change of fermented substrate, tunning respectively, to understand real-time fermentation situation.Establish the concrete processing condition and the requirement of the batch fermentation and the two kinds of various processes of continuously fermenting on the basis of above-mentioned test, propose the mode of production under the different demands, test-results sees table 1 and table 2 for details.
Microbial growth situation under the table 1 different fermentations condition
| Fermentation condition | The microbe colony number (individual/mL) | ||||
| Subtilis | Pseudomonas stutzeri | Aquatic candiyeast | Saccharomyces globosus | ||
| pH | 5.6 7.0 8.5 | 8×10 82×10 96×10 7 | 2×10 95×10 92×10 8 | 1×10 93×10 85×10 7 | 3×10 85×10 72×10 7 |
| Inoculum size % | 5 10 20 | 1×10 92×10 92×10 9 | 1×10 95×10 95×10 9 | 8×10 81×10 91×10 9 | 7×10 73×10 83×10 8 |
| Culture temperature ℃ | 25 30 35 | 6×10 82×10 91×10 9 | 7×10 85×10 92×10 9 | 7×10 81×10 91×10 9 | 6×10 73×10 83×10 8 |
| Incubation time h | 12 18 24 | 5×10 81×10 92×10 9 | 8×10 85×10 95×10 9 | 8×10 81×10 91×10 9 | 2×10 83×10 83×10 8 |
| Air flow % | 30 50 100 | 5×10 76×10 82×10 9 | 3×10 79×10 85×10 9 | 4×10 87×10 81×10 9 | 5×10 67×10 73×10 8 |
As shown in Table 1, subtilis and the suitableeest initial pH of Pseudomonas stutzeri fermentation are 7.0, and inoculum size is 10%, and culture temperature is 30 ℃, and incubation time is 18-24h, and air flow is 100%; Aquatic candiyeast and the suitableeest initial pH of Saccharomyces globosus fermentation are 5.6, and inoculum size is 10%, and culture temperature is 30 ℃, and incubation time is 12-18h, and air flow is 100%.Under the optimal culture conditions of each bacterial classification, the 4 bacterial classifications 6h after fermentation, 9h, 12h, 15h, 18h and 24h respectively carry out analytical results to the variation of the output of fermented substrate and tunning and see table 3 for details.
The fermentation of table 2 different fermentations time is dynamic
| Fermentation time | Subtilis | Pseudomonas stutzeri | Aquatic candiyeast | Saccharomyces globosus | |
| pH | 6h 9h 12h 15h 18h 24h | 5.8 4.6 3.8 4.2 4.4 4.3 | 5.5 4.3 3.6 3.8 4.0 4.2 | 4.2 2.8 3.2 3.5 3.6 3.8 | 4.5 3.0 3.2 3.2 3.5 3.9 |
| Pol | 6h 9h 12h 15h 18h 24h | 3.0 2.5 1.5 1.2 1.0 1.0 | 2.8 2.2 1.3 1.2 1.0 1.0 | 2.5 1.5 1.2 1.2 1.0 1.0 | 2.8 2.0 1.5 1.2 1.2 1.0 |
| Total nitrogen ‰ | 6h 9h 12h 15h 18h 24h | 0.2 0.1 0.08 0.05 0.02 0.02 | 0.18 0.09 0.07 0.03 0.02 0.01 | 0.15 0.07 0.06 0.03 0.02 0.01 | 0.18 0.09 0.07 0.04 0.02 0.02 |
| Bacterium colony several/mL | 6h 9h 12h 15h 18h 24h | 6×10 6 8×10 7 5×10 8 8×10 8 1×10 9 2×10 9 | 8×10 6 1×10 8 8×10 8 1×10 9 3×10 9 5×10 9 | 5×10 7 6×10 8 8×10 8 1×10 9 1×10 9 1×10 9 | 8×10 6 4×10 7 1×10 8 3×10 8 3×10 8 3×10 8 |
By table 2 as seen, in the process of cultivating, the growth of the reduction of pH and the consumption of substrate and bacterium amount has direct relation, during rapid reduction of pH just the bacterium amount begin stage of logarithmic growth, carbon source and nitrogenous source also begin by a large amount of consumption.On the basis of above-mentioned test, establish the batch fermentation and the two kinds of different production technique of continuously fermenting.Batch fermentation is: produce the feed supplement to 9-12h, 3-6h discharging after the feed supplement.Continuously ferment for: produce to 12-15h discharging 70%, continue to cultivate by original pattern after replenishing 70% substratum, the cultured continuously that so moves in circles 3 days is as one-period.
Embodiment 4: the preparation of compound micro-ecological preparation
1) shakes a bottle rejuvenation for a short time
Use transfering loop respectively under aseptic technique on the inclined-plane of 4 kinds of bacterium, be inoculated in the shaking for a short time in the bottle of the 150mL that contains the 30mL seed culture medium, in shaking table, carry out pure culture, condition is 30 ℃ of culture temperature, incubation time 24h, shaking speed 150r/min, the microscopy bacterium is pollution-free and form is normal, and every kind of bacterium viable count is not less than 10
8/ mL;
2) shake the bottle amplification greatly
Cultivate 4 kinds of qualified bacterium through microscopy and under aseptic technique, be transferred to shaking greatly in the bottle of the 2500mL that contains the 500mL seed culture medium respectively shaking for a short time in the bottle, in shaking table, carry out pure culture, condition is 30 ℃ of culture temperature, incubation time 24h, shaking speed 170r/min, the microscopy bacterium is pollution-free and form is normal, and every kind of bacterium viable count is not less than 10
8/ mL;
3) seed tank culture base preparation
The fermention medium of preparing subtilis, Pseudomonas stutzeri, aquatic candiyeast and Saccharomyces globosus respectively is 80L, join then in 4 seed fermentation jars, carry out the steam high-temperature sterilization of seeding tank and associated conduit, condition is 125 (± 5) ℃, time is 30min, the sterilization back is cooled to about 30 ℃ with recirculated cooling water, and the fermentor tank pressurize is to 0.1MPa;
4) seeding tank fermentation
Insert respectively through check to 4 seeding tanks by the inoculation valve rapidly under the flame protection and cultivate qualified seed liquor, inoculum size is 15%, and fermentation culture conditions is air flow 4m
3/ h, culture temperature 30 (± 1) ℃, incubation time 24h, the fermentor tank pressurize is to 0.1MPa, stirrer rotating speed 150r/min, tank pressure 0.10MPa, the pH value is normal fermentation state value about 3.5; The microscopy bacterium is pollution-free and form is normal, and every kind of bacterium viable count is not less than 10
8/ mL.
5) second order fermentation jar medium preparation
Prepare the fermention medium 800L of subtilis, Pseudomonas stutzeri, aquatic candiyeast and Saccharomyces globosus respectively, 800L and 1600L, 1600L, 4 second order fermentation jars and associated conduit are carried out the steam high-temperature sterilization, condition is 125 (± 5) ℃, time is 30min, and the sterilization back is cooled to about 30 ℃ with recirculated cooling water, and the fermentor tank pressurize is to 0.1MPa;
6) second order fermentation jar large scale fermentation
Pressure by reducing by 4 second order fermentation jars and increase the pressure of 4 seeding tanks, the seed liquor of 4 seeding tanks that will be through being up to the standards moves in the corresponding second order fermentation jar carries out multiplication culture, and fermentation culture conditions is air flow 4m
3/ h, culture temperature 30 (± 1) ℃, incubation time 24h, the fermentor tank pressurize is to 0.1MPa, stirrer rotating speed 170r/min, tank pressure 0.10MPa, the pH value is normal fermentation state value about 3.5; The microscopy bacterium is pollution-free and form is normal, and every kind of bacterium viable count is not less than 10
8/ mL.
7) finished product can
The bacterium liquid of second order fermentation jar that will be through being up to the standards carries out can after moving in proportion and carrying out aeration-agitation mixing 10h in the sterilized storage tank then between sterile filling.
Embodiment 5: the combination of compound micro-ecological preparation and to the influence of cultivating pool bait microorganism
With aquatic candiyeast, Saccharomyces globosus, subtilis, Pseudomonas stutzeri by 0.8: 1.2: 2.3: 1.5 are optimized test to the cultivating pool food organism after being combined into compound micro-ecological preparation, the result shows, form in the structure of algae, the test pool is after July, promptly progressively develop into 42 kinds by original 34 kinds and 35 kinds, its sociales are mainly diatom, green alga, Euglena and part blue-green algae, wherein test No. 2 pools and are mainly diatom and green alga.And the contrast pool from start to finish blue-green algae be the remarkable advantages kind, 7～September, the blue-green alga bloom population was more outstanding and serious on the occasion of high temperature season.The algae population is promptly developed into 21 kinds by original 34 kinds.Show from the composition of algae structure, compound micro-ecological preparation is obvious to the restraining effect of deleterious blue-green alga bloom population, the algae structure is had regulatory function, and its result makes the blue-green algae on the test pool be converted into subordinate species by advantage, and changes diversity into by the unicity that algae is formed.Zooplankton also has identical rule, promptly test the pool and be mainly protozoon and large cyclop, and the contrast pool is mainly wheel animalcule.According to Shannon biodiversity index evaluating water quality environment, compound micro-ecological preparation not only has optimizational function to composition and the quantity of pond food organism, and plays an important role to alleviating the pond water quality organic contamination.From the measurement result of pond algae bio amount, compound micro-ecological preparation has the obvious control effect to the blue-green algae biomass, and other algae bio amount is had the promotion growth and breeding, improves the effect of biomass.The test pool is 75% to No. 1 pool of the inverse amplification factor of blue-green algae test, and test No. 2 pools is 86%, is 100～200% to the rate of increase of other algae bio amount, and this increases productivity, increases output to increase palatability food organism, has important effect.Also demonstrate same rule from the measurement result of zooplankton biomass.
Embodiment 6: the application of compound micro-ecological preparation in different breeding ecology
With aquatic candiyeast, Saccharomyces globosus, subtilis, Pseudomonas stutzeri by 0.5: 1.5: 2.0: 1.5 are combined into behind the compound micro-ecological preparation and use in the different aquaculture wateies, and experimental result is as follows:
(1) no matter compound micro-ecological preparation is to indoor little water body, and still to outdoor large-area aquaculture water, it all has stronger water quality purification function.Compound micro-ecological preparation can be stablized the pH value. and improve the dissolved oxygen amount of water body, reduce ammonia nitrogen in the water body. nitrite. the content of sulfide and COD etc., keep the benign eubiosis of water body.
(2) the compound micro-ecological preparation cell individual less, nutritious, be easy to digestion, be applied to Macrobrachium rosenbergii breeding, transition bait as zoea open-mouthed bait and post-larva, can promote larval metamorphosis to grow, strengthen the mobility of the young, improve the surviving rate of the young, can not only increase the output of seedling (output of seedling improves 55.9% than control group) of unit of water body, and can make the individual specification of shrimp seedling more neat, be preferable nutritional additive of prawn seed-rearing phase.
(3) adopt compound micro-ecological preparation 40～60mL/40m
3The dosage full pool spilling head, plentiful the degree coefficient of the raising to some extent of the surviving rate of Penaeus japonicus juvenile, relative growth rate and juvenile prawn, also can optimize the water quality of aquaculture water simultaneously, stablize the pH value of water body and make DO content improve 30%, reduce ammonia nitrogen, nitrite and COD in the water, for the existence of aquatic living things provides the ecotope of a healthy and beneficial, to the disease control of aquaculture water with keep the eubiosis in waters to play an important role.Adopt the method for full pool spilling head easy to use, respond well, be worth in large-area breeding production, promoting the use of.
Embodiment 7: compound micro-ecological preparation is to the organic biological degradation of aquaculture water
By being combined into compound micro-ecological preparation organism in the purifying aquatic water afterwards at 2: 1: 2, test-results sees Table 3 with candiyeast, subtilis, pseudomonas.
Table 3 compound strain is to organic biological degradation in the aquaculture water
| Bacterial strain ratio 1 #∶2 #∶3 # | Subject range | Organic cod MnConcentration (mg/L) | Inoculum size (%) | Action time (h) | Degradation rate (%) | ||
| pH | DO(mg/L) | Saltiness (G/L) | |||||
| 1∶1∶1 1∶2∶1 1∶1∶2 1∶2∶2 2∶1∶1 2∶1∶2 2∶2∶1 | 6.5～7.5 6.5～7.5 5.6～7.5 5.6～7.5 6.5～8.5 5.6～8.5 6.5～8.5 | ≥4.0 ≥4.0 ≥4.0 ≥4.0 ≥2.5 ≥2.5 ≥2.5 | 0～25 0～35 0～35 0～35 0～25 0～35 0～35 | 100 100 100 100 100 100 100 | 0.05 0.05 0.05 0.05 0.05 0.05 0.05 | 48 48 48 48 48 48 48 | 30～42 48～58 52～67 48～60 42～56 82～91 69～80 |
As can be seen from Table 3, the subject range of compound micro-ecological preparation is the widest, and the degradation rate of 48h is also maximum, can reach 82-91%, the most degradation rate of its most degradation rate 30%, 50% and 70% during than single bacterial strain effect 48h improves 200%, 80% and 30% respectively.When degradation rate improves, can make the inoculum size of candiyeast and genus bacillus drop to 0.05% by 2 ‰, rate of descent is 97.5%.The organic contamination that is combined as aquaculture system of this ratio purifies provides the biodegradation method of comparatively optimizing, and has certain practical significance.
Claims (11)
1, a kind of aquaculture and scenery water body compound micro-ecological preparation, it is characterized in that with aquatic candiyeast, Saccharomyces globosus, subtilis and Pseudomonas stutzeri be active ingredient, press 0.5-1.7 by four kinds of bacterium and substratum separately: the mass ratio of 0.0-1.5: 1.0-3.0: 1.0-2.5 is formulated.
2, aquaculture and scenery water body compound micro-ecological preparation according to claim 1 is characterized in that the viable bacteria number average of every kind of bacterium is not less than 10
8/ mL.
3, aquaculture and scenery water body compound micro-ecological preparation according to claim 1 is characterized in that degrading and transforming the eutrophication material of aquaculture and landscape water body, suppress the blue-green algae growth, purify water.
4, aquaculture and scenery water body compound micro-ecological preparation as claimed in claim 1 or 2 is characterized in that the seed of each bacterial classification and fermention medium by the formulated aqueous solution of following material, the nutrient media components scope following (unit: g/L):
Aquatic candiyeast: molasses 5-15, peptone 0-1, glucose 5-15, ammonia chloride 1-4, yeast extract paste 0.1-0.5, potassium primary phosphate 0.2-1.0, dipotassium hydrogen phosphate 0.5-1.5, sal epsom 0.3-0.8, sodium-chlor 0.4-0.7, ferrous sulfate (0.001-0.005) * 10
-3, water 1L;
Saccharomyces globosus: molasses 6-14, peptone 0-2, glucose 7-16, ammonia chloride 1.5-3.0, yeast extract paste 0.2-0.7, potassium primary phosphate 0.3-0.8, dipotassium hydrogen phosphate 0.4-1.5, sal epsom 0.35-0.7, sodium-chlor 0.3-0.9, ferrous sulfate (0.002-0.005) * 10
-3, water 1L;
Subtilis: molasses 2-13, peptone 1-6, glucose 2-8, ammonia chloride 0.5-3.0, yeast extract paste 0.15-0.45, potassium primary phosphate 0.25-0.6, dipotassium hydrogen phosphate 0.6-0.9, sal epsom 0.2-1.0, sodium-chlor 0.2-1.3, ferrous sulfate (0.001-0.004) * 10
-3, water 1L;
Pseudomonas stutzeri: molasses 6-17, peptone 1.5-4, glucose 3-10, ammonia chloride 0.7-2.5, yeast extract paste 0.21-0.7, potassium primary phosphate 0.1-0.7, dipotassium hydrogen phosphate 0.3-1.2, sal epsom 0.4-0.9, sodium-chlor 0.1-0.8, ferrous sulfate (0.002-0.007) * 10
-3, water 1L.
5, the preparation method of any described aquaculture of claim 1 to 4 and scenery water body compound micro-ecological preparation is characterized in that comprising the steps:
1) shakes a bottle rejuvenation for a short time
Use transfering loop respectively under aseptic technique on the inclined-plane of 4 kinds of bacterium, be inoculated in and contain the shaking for a short time in the bottle of seed culture medium, in shaking table, carry out pure culture respectively;
2) shake the bottle amplification greatly
Will shake for a short time cultivate in the bottle and contain the shaking greatly in the bottle of seed culture medium, in shaking table, carry out pure culture after 4 kinds of bacterium that are up to the standards are transferred under aseptic technique respectively;
3) preparation of seeding tank fermention medium
Preparation seeding tank fermention medium is also put into seeding tank, and the steam high-temperature sterilization postcooling of seeding tank and associated conduit is to temperature required;
4) seeding tank fermentation
Insert respectively through check to 4 seeding tanks by the inoculation valve rapidly under the flame protection and cultivate qualified seed liquor, inoculum size is 10%-20%;
5) second order fermentation jar medium preparation
Preparation second order fermentation jar substratum is also put into the second order fermentation jar, and second order fermentation jar and associated conduit are carried out steam high-temperature sterilization postcooling to temperature required;
6) second order fermentation jar large scale fermentation
By pressure that reduces the second order fermentation jar and the pressure that increases seeding tank, the seed liquor of 4 seeding tanks that will be through being up to the standards moves in the second order fermentation jar carries out multiplication culture, and the culture transferring amount is 10%-20%
7) finished product can
The bacterium liquid of second order fermentation jar that will be through being up to the standards carries out can after moving in proportion and carrying out aeration-agitation mixing 5-12h in the sterilized storage tank then between sterile filling.
6, preparation method as claimed in claim 5 is characterized in that step 1) and 2) culture condition be temperature 27-37 ℃, rotating speed 110-220r/min, incubation time are 17-24h.
7, preparation method as claimed in claim 5, the temperature condition that it is characterized in that steam high-temperature sterilization in step 3) and the step 5) is, 121 ℃ of conditions continue to stop behind the 20-30min, to the fermentor tank pressurize between the 0.02-0.18MPa and be cooled between 27-37 ℃.
8, preparation method as claimed in claim 5 is characterized in that the fermentation condition of step 4) is: temperature 27-37 ℃, stirrer rotating speed 110-220r/min, incubation time are 17-24h, air flow 4m
3/ h, tank pressure 0.02-0.18MPa, the pH value is 3.0-7.0.
9, preparation method as claimed in claim 5 is characterized in that the fermentation condition in the step 6): temperature 27-37 ℃, and stirrer rotating speed 110-220r/min, incubation time are 17-24h, air flow 4m
3/ h, the tank pressure hold-in range is 0.02-0.18MPa.
10, preparation method as claimed in claim 5 is characterized in that every kind of bacterium viable count is not less than 10 in the finished product that step 7) makes
8/ mL.
11, preparation method as claimed in claim 5 is characterized in that step 1) and step 2) in the consumption of substratum divide and to be 30mL and 500mL in addition.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105087410A (en) * | 2014-05-11 | 2015-11-25 | 无锡中科活力生物技术有限公司 | Novel aquaculture probiotics, and production method and use thereof |
| CN106277579A (en) * | 2016-08-15 | 2017-01-04 | 吴小慧 | The deep treatment method of cultivation factory sewage |
| CN106630189A (en) * | 2016-12-15 | 2017-05-10 | 雷州市汇大生物科技有限公司 | Liquid marine red yeast water quality improver for aquaculture and preparation method thereof |
| CN107986456A (en) * | 2017-12-14 | 2018-05-04 | 泰州永达生物科技有限公司 | A kind of prevention and suppressing method applied to water body cyanobacteria |
| CN110769687A (en) * | 2017-04-20 | 2020-02-07 | 洛克斯Ip有限责任公司 | Cost effective compositions and methods for enhancing aquaculture and ornamental fish feeding |
| CN113528386A (en) * | 2021-07-14 | 2021-10-22 | 大连金砣水产食品有限公司 | Preparation method of aquatic product composite microecological preparation |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105087410A (en) * | 2014-05-11 | 2015-11-25 | 无锡中科活力生物技术有限公司 | Novel aquaculture probiotics, and production method and use thereof |
| CN105087410B (en) * | 2014-05-11 | 2018-10-02 | 无锡中科活力生物技术有限公司 | A kind of novel aquaculture probiotics and its production method and purposes |
| CN106277579A (en) * | 2016-08-15 | 2017-01-04 | 吴小慧 | The deep treatment method of cultivation factory sewage |
| CN106630189A (en) * | 2016-12-15 | 2017-05-10 | 雷州市汇大生物科技有限公司 | Liquid marine red yeast water quality improver for aquaculture and preparation method thereof |
| CN110769687A (en) * | 2017-04-20 | 2020-02-07 | 洛克斯Ip有限责任公司 | Cost effective compositions and methods for enhancing aquaculture and ornamental fish feeding |
| CN107986456A (en) * | 2017-12-14 | 2018-05-04 | 泰州永达生物科技有限公司 | A kind of prevention and suppressing method applied to water body cyanobacteria |
| CN113528386A (en) * | 2021-07-14 | 2021-10-22 | 大连金砣水产食品有限公司 | Preparation method of aquatic product composite microecological preparation |
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| CN100467584C (en) | 2009-03-11 |
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