CN101595943A - A kind of production method of additive of bait for photosynthetic bacteria aquaculture - Google Patents
A kind of production method of additive of bait for photosynthetic bacteria aquaculture Download PDFInfo
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- CN101595943A CN101595943A CNA2009100395240A CN200910039524A CN101595943A CN 101595943 A CN101595943 A CN 101595943A CN A2009100395240 A CNA2009100395240 A CN A2009100395240A CN 200910039524 A CN200910039524 A CN 200910039524A CN 101595943 A CN101595943 A CN 101595943A
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Abstract
The present invention relates to a kind of production method of additive of bait for photosynthetic bacteria aquaculture.This method is taked following steps: 1. bacterial classification enlarges cultivation: bacterial classification is enlarged cultivate keynote pH6.0-8.5, pack into vial and carry out high-temperature sterilization, the 5%-40% that presses the bacterium culture medium volume inserts the mixed culture of Rhodopseudomonas palustris and two kinds of photosynthetic bacterias of Spherical red antibacterial.15 ℃-35 ℃ of temperature, intensity of illumination is that anaerobism was cultivated 3-4 days under the 1000lux-3000lux condition, bacteria culture fluid; 2. fermenting and producing: pack into fermentation medium in the fermentation tank and carry out high-temperature sterilization, insert bacteria culture fluid by the inoculum concentration of the 5%-40% of fermenting and producing culture volume.At temperature 28-30 ℃; Mixing speed 150-300 rev/min; Tank pressure keeps 0.02-0.05MPa; Intensity of illumination is 2000-3000lux condition bottom fermentation 6 days, additive of bait for photosynthetic bacteria aquaculture.The present invention compared with prior art has the advantage of constant product quality, functional diversities.
Description
Technical field
The present invention relates to microorganism field, relate in particular to a kind of production method of additive of bait for photosynthetic bacteria aquaculture.
Background technology
Along with the high speed development of China worker, agricultural, the raising of living standards of the people, the direct discharging of a large amount of undressed workers, agricultural production waste water and sanitary sewage; Add the raising of China's aquaculture intensification degree, make China culture water pollution and be on the rise.The breed aquatic animal disease of Yin Faing rolls up thus, and frequent the generation.And blindly a large amount of frequent fishing (beast) medicine, antibiotic, hormones etc. of using make the Zhi amount safety of aquatic products be subjected to serious threat.In order to protect the fishery ecological environment, improve aquatic products Zhi amount and output, promote sustainable fisheries industry development, use photosynthetic bacteria probiotics control fishery ecological environmental pollution and destruction, control aquatic products disease, significant undoubtedly.Chinese invention patent numbers 94106402.6 discloses the technology that a kind of name is called " animal feed additive and the production method thereof of utilizing micro-organisms ".This animal feed additive is a kind of liquid preparation of producing with photosynthetic bacteria, also use in aquaculture, but because its raw materials for production adopt the amidin of 10-40%, nutrient is not comprehensive, be difficult to satisfy the needs of photosynthetic bacterium growth to a great number of elements such as nitrogenous source, phosphorus source, sulphur sources, thereby undergrowth.Simultaneously, the amidin that this patent is used adopts the mode of self-souring to obtain acid fermentation liquid after saccharification, owing to the acidifying bacterial classification is not artificially controlled, thereby the difficult quality guarantee of acid fermentation liquid.And sterilized is handled before inoculation photosynthetic bacteria bacterial classification, so contain a large amount of heterotrophs in the acid fermentation liquid, these heterotrophs can suppress the growth of photosynthetic bacteria, thereby cause the severe contamination of product, pollution rate can reach more than 80%, thereby causes the instability of application effect.
Summary of the invention
Goal of the invention of the present invention is in order to overcome the deficiency of prior art, the production method of the additive of bait for photosynthetic bacteria aquaculture of a kind of steady quality and functional diversities to be provided.
Goal of the invention of the present invention is to realize like this; The production method of this additive of bait for photosynthetic bacteria aquaculture is taked following steps:
1, bacterial classification enlarges cultivation
Produce bacterial classification: adopt culture presevation number be M209077 Rhodopseudomonas palustris (H1) (Rhodopeudomonas palustris) and culture presevation number for 1: 1 the mixed cultures of two kinds of photosynthetic bacterias of the Spherical red antibacterial of M209078 (H2) (Rhodobacter sphaeroides) as the production bacterial classification.This two strains bacterial classification is deposited in " the Chinese typical culture collection center " of Wuhan University on April 27th, 2009.
Bacterial classification is enlarged the following composition of culture medium: natrium malicum 1.0 grams; Sodium acetate 1.0 grams; Ammonium chloride 1.0 grams; Potassium dihydrogen phosphate 1.0 grams; Calcium chloride 0.1 gram; Sodium acid carbonate 3.0 grams; Magnesium chloride 0.5 gram; Trace element solution 1.0ml; Vitamin solution 1.0ml; Yeast extract 0.5 gram is dissolved in respectively in 1.0 liters of distilled water.Yong Qing Yangization Na transfers in the pH6.0-8.5 scope, and preferably pH is 6.8-7.2, the vial of packing into.121 ℃ of sterilizations 30 minutes.When treating that culture medium is cooled to 30-35 ℃, press the 5%-40% of bacterium culture medium volume, preferably the inoculum concentration of 10%-20% inserts the mixed culture of above-mentioned two kinds of photosynthetic bacterias.In 15 ℃ of-35 ℃ of scopes of temperature, best 28 ℃-30 ℃, intensity of illumination is that anaerobism was cultivated 3-4 days under the 10001ux-3000lux condition, and bacteria containing amount reaches 5X10
8-10X10
8Cfu/ml, red bacteria culture fluid.
Trace element solution is formed: four water frerrous chlorides, 1.8 grams; CoCL2 0.25 gram; Six water nickel chlorides, 0.01 gram; Copper chloride dihydrate 0.01 gram; Tetrahydrate manganese chloride 0.7 gram; Zinc chloride 0.1 gram; Boric acid 0.5 gram; Sodium Molybdate Dihydrate 0.03 gram; Sodium selenite (Na2SeO3) pentahydrate 0.01 gram; 1.0 liters of distilled water.
Vitamin solution is formed: biotin 0.1 gram; Nicotinic acid 0.35 gram; Thiamine hydrochloride 0.3 gram; Para-Aminobenzoic 0.2 gram; Hydrochloric acid pyridoxamine 0.1 gram; Calcium pantothenate 0.1 gram; Cobalamin 0.05 gram; 1.0 liters of distilled water.
2, fermenting and producing
(1) fermentation medium is formed: malic acid 1.0 grams; Sodium acetate 1.5 grams; Sodium propionate 0.5 gram; Yeast extract 1.0 grams; Ammonium sulfate 1.3 grams; Epsom salt 0.2 gram; Calcium chloride dihydrate 0.07 gram; Ironic citrate 0.01 gram; Ethylenediamine tetra-acetic acid (EDTA) 0.02 gram; Potassium dihydrogen phosphate 0.6 gram; Dipotassium hydrogen phosphate 0.9 gram; Trace element solution 1.0ml; Vitamin solution 7.5ml; 1.0 liters in running water; PH 6.8-7.2.
Wherein:
Trace element solution: ironic citrate 3 grams; Manganese sulfate monohydrate 0.02 gram; Boric acid 0.01 gram; Cupric sulfate pentahydrate 0.01 gram; Four water ammonium molybdates, 0.02 gram; Zinc sulfate 0.01 gram; Ethylenediamine tetra-acetic acid (EDTA) 0.5 gram; Calcium chloride dihydrate 0.2 gram; 1.0 liters of distilled water.
Vitamin solution: nicotinic acid 0.2 gram; Niacinamide 0.2 gram; Thiamine salt hydrochlorate 0.4 gram; Biotin 0.08 gram; 1.0 liters of distilled water.
(2) production process and condition control: in full-automatic photosynthetic microorganism fermentation tank, by above-mentioned composition preparation fermentation medium.Sterilized 60 minutes for 121 ℃.When treating that culture medium is cooled to 30 ℃-35 ℃, by the 5%-40% of fermenting and producing culture volume, the inoculum concentration that is preferably 10%-20% inserts bacteria culture fluid.At temperature 28-30 ℃; Mixing speed 150-300 rev/min; Keep tank pressure with filtrated air, tank pressure keeps 0.02-0.05MPa; Intensity of illumination initial stage 2000lux, mid-term 2500lux, later stage 3000lux condition bottom fermentation 6 days, the rufous additive of bait for photosynthetic bacteria aquaculture.Include living bacteria count 5X10
9-6X10
9Cfu/ml; PH7.0-8.5; Assorted bacterium rate is less than 2%.
The present invention compared with prior art has the following advantages:
1, the additive of bait steady quality of producing with the present invention.
The present invention adopts pure chemical reagent to make raw materials for production, and Zhi amount and quantity obtain strict control, nutritive element balance, comprehensive.Can satisfy the demand of photosynthetic bacterium growth breeding to carbon source, nitrogenous source, phosphorus source, sulphur source and various trace elements.In addition, before inoculation photosynthetic bacteria mixed culture, adopt 121 ℃ of sterilizations 60 minutes, killed vegetative cell, brood cell and the mold spore of all assorted bacterium in the fermenting and producing culture medium.Adopt filtrated air to keep tank pressure in process of production again, make photosynthetic bacteria growth and breeding under the condition of pure culture fully.Every batch of product includes living bacteria count 5X10
9-6X10
9Cfu/ml, assorted bacterium rate is less than 2%, thus the Zhi that has guaranteed additive of bait measures and application effect.
2, the additive of bait of producing with the present invention has following multiple efficacies:
This additive of bait for photosynthetic bacteria aquaculture is the bioactive additive that integrates nutrition and health care, and its effect is equivalent to feed addictive ten growth accelerators ten antiviral agents ten water purification agents.
1), provides nutrition, promotion growth, raising bait utilization, increasing both production and income
The main component photosynthetic bacteria cell of this additive of bait contains profuse albumen Zhi, B family vitamin, carotenoids, trace element, also is rich in nucleic acid, ubiquinone
10With somatomedin etc.In eel culture, in bait,, on average increase by 8.19% than conventional cultivation output by behind 1% this additive of interpolation; Feed coefficient on average descends 12.6%; Growth cycle has shortened 15 days.In the California perch was cultured, by behind 1% this additive of interpolation, than conventional cultivation, output increased by 7.78% in bait; Feed coefficient descends 5.56%; Growth cycle shortens 28 days.
2), purify water, safeguard the water ecology balance
In eel culture, every water Zhi index of measuring before not using this additive is: pH6.7; Dissolved oxygen 5.5mg/L; Ammonia nitrogen 1.132mg/L; Nitrite 0.413mg/L; Hydrogen sulfide 0.003mg/L.By 1kg/ mu * rice this additive of splashing, water Zhi testing result is as follows after 5 days: pH7.6 (becoming alkalescent by faintly acid) then; Dissolved oxygen 7.8mg/L (increasing 41.8%); Ammonia nitrogen 0.465mg/L (reducing by 58.9%); Nitrite 0.101mg/L (reducing by 75.5%); Hydrogen sulfide 0mg/L (degraded 100%).Every index all is better than " fresh water fishery water quality is cultured standard ".In California perch and Penaeus Vannmei breeding process, adhere to per 15 days by 1kg/ mu * rice this additive of splashing, sampling and measuring water Zhi weekly, every index all is better than " fresh water fishery is cultured water Zhi standard ", and useful zooplankter quantity such as wheel animalcule, branch angle worm obviously increases in the water body.
3), degraded medicament residue and water pollutant
(1), this aquaculture additive is very obvious to multiple aquaculture medicament residue degradation effect.Can reach 84.6% to 120 hours the degradation rate of water body that contains the 10mg/L malachite green; To containing the water body of 10mg/L furazolidone, 72 hours degradation rate reaches 100%.
(2), in outlet eel live body detects, find that the fish body includes nitrofuran metabolite 3.15mg/kg, add this additive by 3% subsequently and in bait, feed, after 10 days once more testing result be: can't detect, the vivo degradation rate reaches 100%, reaches export standard.
(3), this additive contains the enzyme and the Zhi grain of multiple degradation of contaminant, except each type organic of degradable, also thing Zhi of the many difficult for biological degradation of degradable such as Halogen can reach 96.4% for aromatic compound, amine, alkanes etc. to the chlorobenzene compounds clearance that is widely used in the industrial and agricultural production.
4), fixing heavy metal in the water body, reduce the harm of heavy metal to aquatic animal; And reduce aquatic livestock heavy metal of body content, ensure food safety.This additive can be converted into water-soluble poisonous and harmful heavy metallic water-fast metal sulfide, thereby has avoided the harm of heavy metal to aquatic animal to grow; Also reduce the enrichment of heavy metal in the aquiculture animal body, reduced heavy metal of body content.Test shows: this additive is to Pb
2+, As
5+, As
3+, Cd
2 +, Cr
2+, Hg
2+, Cu
2+Etc. various heavy very strong resistance is arranged.To initial concentration is the Cd of 10-25mg/L
2+, 36 hours clearance can be more than 85%.To concentration is the Pb of 100mg/L
2+Clearance can reach 95% in 24 hours.
5), disease resistance, improve immunity and survival rate
This additive contains effectively passivation water body virus of antiviral ingredients, reduces the generation of virosis.Multiple aquaculture pathogens such as Aeromonas sobria, pseudomonas putida also there is better inhibitory action.This additive can promote the growth of animal immune organ in addition, effectively improves animal immunizing power.Eel culture experiment shows, use this additive after, the fish disease loss reduces 15%, survival rate improves 4.26%; California perch culture experiment shows, uses diseases such as the common fin rot of California perch behind this additive, mashed body disease, fish sore not take place; Survival rate improves 3.19%; Penaeus Vannmei high-density breeding experiment shows, use this additive after, do not re-use the fishing medicine substantially, survival rate is up to more than 95%.
6), improve the quality of aquatic products
(1), in eel culture, use this additive after, the adult fish size is evenly neat, color crow profit gloss, meat Zhi is good, mouthfeel is good.The live body examining report shows that antibiotics, sulfamido, BHC class, DDT class, steroids, chrysanthemum ester class and all kinds of poisonous and harmful heavy metallics etc. are and do not detect.
(2), in the California perch is cultured, use this additive after, the fish body is neat, fish scale is glossy especially, and is fresh and alive vigorous, meat is abundant soft.The live body examining report shows that antibiotics, sulfamido, BHC class, DDT class, steroids, chrysanthemum ester class and all kinds of poisonous and harmful heavy metallics etc. are and do not detect.
The specific embodiment
The present invention is further illustrated below in conjunction with specific embodiment:
Embodiment 1
Step 1: put vial after the various compositions that above-mentioned bacterial classification is enlarged culture medium dissolve in distilled water, transfer pH6.0 with NaOH, 121 ℃ of sterilizations 30 minutes, when being cooled to 30 ℃, the Rhodopseudomonas palustris and the Spherical red antibacterial mixed culture of bacterium culture medium volume 5% gone in inoculation, is 15 ℃ in temperature, and intensity of illumination is that anaerobism was cultivated 3 days under the 1000lux condition, promptly get the bacteria culture fluid of rufous, bacteria containing amount reaches 5X10
8Cfu/ml.
Step 2: put in the full-automatic photosynthetic microorganism fermentation tank after above-mentioned fermentation dissolved in running water with the various compositions of culture medium, 121 ℃ of sterilizations 60 minutes, when treating that culture medium is cooled to 30 ℃, by the inoculum concentration access bacteria culture fluid of fermenting and producing culture volume 5%.In temperature is 30 ℃; 150 rev/mins of mixing speeds; Use the filtrated air pressurize, tank pressure remains 0.02MPa; The intensity of illumination initial stage is 2000lux, and be 2500lux mid-term, and the later stage is 3000lux condition bottom fermentation 6 days, promptly gets the rufous additive of bait for photosynthetic bacteria aquaculture, includes living bacteria count 6X10
9Cfu/ml, assorted bacterium rate is less than 2%.
Embodiment 2
Step 1: will put vial into after the dissolving in the various composition distilled water of above-mentioned bacterial classification expansion culture medium, transfer pH8.5 with NaOH, 121 ℃ of sterilizations 30 minutes, when being cooled to 35 ℃, the Rhodopseudomonas palustris and the Spherical red antibacterial mixed culture of bacterium culture medium volume 20% gone in inoculation, is 35 ℃ in temperature, and intensity of illumination is that anaerobism was cultivated 4 days under the 3000lux condition, promptly get the bacteria culture fluid of rufous, bacteria containing amount reaches 8X10
8Cfu/ml.
Step 2: put in the full-automatic photosynthetic microorganism fermentation tank after above-mentioned fermentation dissolved in running water with the various compositions of culture medium, 121 ℃ of sterilizations 60 minutes, when treating that culture medium is cooled to 35 ℃, by the inoculum concentration access bacteria culture fluid of fermenting and producing culture volume 40%.In temperature is 30 ℃; 300 rev/mins of mixing speeds; Use the filtrated air pressurize, tank pressure remains 0.05MPa; The intensity of illumination initial stage is 2000lux, and be 2500lux mid-term, and the later stage is 3000lux condition bottom fermentation 6 days, promptly gets the rufous additive of bait for photosynthetic bacteria aquaculture, includes living bacteria count 5.8X10
9Cfu/ml, assorted bacterium rate is less than 2%.
Embodiment 3
Step 1: put vial after the various compositions that above-mentioned bacterial classification is enlarged culture medium dissolve in distilled water, transfer pH7.0 with NaOH, 121 ℃ of sterilizations 30 minutes, when being cooled to 32 ℃, the Rhodopseudomonas palustris and the Spherical red antibacterial mixed culture of bacterium culture medium volume 15% gone in inoculation, is 25 ℃ in temperature, and intensity of illumination is that anaerobism was cultivated 4 days under the 2000lux condition, promptly get the bacteria culture fluid of rufous, bacteria containing amount reaches 8X10
8Cfu/ml.
Step 2: put in the full-automatic photosynthetic microorganism fermentation tank after above-mentioned fermentation dissolved in running water with the various compositions of culture medium, 121 ℃ of sterilizations 60 minutes, when treating that culture medium is cooled to 32 ℃, by the inoculum concentration access bacteria culture fluid of fermenting and producing culture volume 15%.In temperature is 29 ℃; 250 rev/mins of mixing speeds; Use the filtrated air pressurize, tank pressure remains 0.04MPa; The intensity of illumination initial stage is 2000lux, and be 2500lux mid-term, and the later stage is 3000lux condition bottom fermentation 6 days, promptly gets the rufous additive of bait for photosynthetic bacteria aquaculture, includes living bacteria count 6X10
9Cfu/ml, assorted bacterium rate is less than 2%.
Claims (7)
1, a kind of production method of additive of bait for photosynthetic bacteria aquaculture is characterized in that taking following steps:
1) bacterial classification enlarges cultivation
The various compositions that bacterial classification enlarged culture medium are dissolved in distilled water respectively, Yong Qing Yangization Na transfers the vial of packing into behind the pH6.0-8.5,121 ℃ of sterilizations 30 minutes, to be cooledly press 5-40% that bacterial classification enlarges culture volume during to 30-35 ℃ and insert two kinds of mixed cultures of 1: 1 of photosynthetic bacteria of Rhodopseudomonas palustris (H1) (Rhodopeudomonas palustris culture presevation number be M209077) and Spherical red antibacterial (H2) (Rhodobacter sphaeroides culture presevation number be M209078), 15 ℃-35 ℃ of temperature, intensity of illumination is that anaerobism was cultivated 3-4 days under the 1000lux-3000lux condition, red bacteria culture fluid;
2) fermenting and producing
The various compositions of an amount of fermenting and producing culture medium are added in the fermentation tank after the dissolving in water fully, sterilized 60 minutes for 121 ℃, when treating that culture medium is cooled to 30-35 ℃, the inoculum concentration of pressing the 5%-40% of fermenting and producing culture volume inserts bacteria culture fluid, 15 °-35 ℃ of temperature; Mixing speed 150-300 rev/min; Use the filtrated air pressurize, tank pressure keeps 0.02-0.05MPa; Intensity of illumination initial stage 2000lux, mid-term 2500lux, later stage 3000lux condition bottom fermentation 6 days, the photosynthetic bacteria aquatic feed additive of rufous.
2, the production method of additive of bait for photosynthetic bacteria aquaculture according to claim 1 is characterized in that described bacterial classification enlarges culture medium and forms: natrium malicum 1.0 grams; Sodium acetate 1.0 grams; Ammonium chloride 1.0 grams; Potassium dihydrogen phosphate 1.0 grams; Calcium chloride 0.1 gram; Sodium acid carbonate 3.0 grams; Magnesium chloride 0.5 gram; Trace element solution 1.0ml; Vitamin solution 1.0ml; Yeast extract 0.5 gram; 1.0 liters of distilled water.
3, the production method of additive of bait for photosynthetic bacteria aquaculture according to claim 1 is characterized in that described bacterial classification enlarges culture medium and transfers pH6.8-7.2 with NaOH.
4, the production method of additive of bait for photosynthetic bacteria aquaculture according to claim 1 is characterized in that it is the 10%-20% that bacterial classification enlarges culture volume that described bacterial classification enlarges the inoculum concentration of cultivating.
5, the production method of additive of bait for photosynthetic bacteria aquaculture according to claim 1 is characterized in that it is 28 ℃-30 ℃ that described bacterial classification enlarges the temperature of cultivating.
6, the production method of additive of bait for photosynthetic bacteria aquaculture according to claim 1 is characterized in that described fermenting and producing culture medium composition: malic acid 1 gram; Sodium acetate 1.5 grams; Sodium propionate 0.5 gram; Yeast extract 1.0 grams; Ammonium sulfate 1.3 grams; Epsom salt 0.2 gram; Calcium chloride dihydrate 0.07 gram; Ironic citrate 0.01 gram; Ethylenediamine tetra-acetic acid (EDTA) 0.02 gram; Potassium dihydrogen phosphate 0.6 gram; Dipotassium hydrogen phosphate 0.9 gram; Trace element solution 1.0ml; Vitamin solution 7.5ml; 1.0 liters of pH 6.8-7.2 of running water.
7, the production method of additive of bait for photosynthetic bacteria aquaculture according to claim 1, the inoculum concentration that it is characterized in that described fermenting and producing is the 10%-20% of fermenting and producing culture volume.
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| CN105543146A (en) * | 2016-02-02 | 2016-05-04 | 田维熙 | Preparation method of photosynthetic bacterium water aqua and special culture medium of photosynthetic bacterium water aqua |
| CN105543146B (en) * | 2016-02-02 | 2018-11-13 | 田维熙 | A kind of preparation method and its special culture media of photosynthetic bacteria aqua |
| CN108220203B (en) * | 2018-03-06 | 2021-03-02 | 上海海洋大学 | Fermentation medium of rhodobacter sphaeroides and application of fermentation medium in fermentation production of rhodobacter sphaeroides |
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| CN108967743A (en) * | 2018-07-13 | 2018-12-11 | 江苏省海洋水产研究所 | It is a kind of for improving the composite preparation and its preparation method and application of content astaxanthin in white leg Shrimp body |
| CN109593676A (en) * | 2018-12-21 | 2019-04-09 | 江苏大学 | A kind of culture medium and preparation method thereof separated for microorganism in sauerkraut fermentation liquid |
| CN109593676B (en) * | 2018-12-21 | 2023-04-07 | 江苏大学 | Culture medium for separating microorganisms from pickled Chinese cabbage fermentation liquor and preparation method thereof |
| CN109929783A (en) * | 2019-04-22 | 2019-06-25 | 广东宏隆生物科技有限公司 | The culture medium and cultural method of Rhodopseudomonas palustris |
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