CN101531977A - Method for adjusting and controlling mixed culture of floating microalgae by bacillus - Google Patents
Method for adjusting and controlling mixed culture of floating microalgae by bacillus Download PDFInfo
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- CN101531977A CN101531977A CN200910038908A CN200910038908A CN101531977A CN 101531977 A CN101531977 A CN 101531977A CN 200910038908 A CN200910038908 A CN 200910038908A CN 200910038908 A CN200910038908 A CN 200910038908A CN 101531977 A CN101531977 A CN 101531977A
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Abstract
The invention discloses a method for adjusting and controlling mixed culture of floating microalgae by bacillus. The microalgae, the dominance degree of which is more than 0.15, is selected and placed into a culture solution to carry out mixed culture, wherein in the culture solution, the concentration of nitrogen is 12 to 16 mg/L, the concentration of phosphorus is 2 to 6 mg/L and the concentration of iron is 0.10 to 0.20 mg/L; during the mixed culture, the Bacillus licheniformis is adopted for adjustment and control; and the culture conditions comprise that the temperature is between 26 and 28 DEG C, the salinity is 12 to 18 per mill, the pH is 7.5 to 8.0, the illumination intensity is 4,000 to 5,000Lx, and the ratio of light to darkness is 12 hours to 12 hours and 14 hours to 10 hours. By adopting the method for carrying out the mixed culture on the microalgae by using the bacillus in a mixed culture system, the growth of the microalgae is balanced, the stability is good, single biological diversity of algae phases in the mixed culture system can be avoided, and extreme advantages or disadvantages on the number of the microalgae can be avoided.
Description
Technical field
The present invention relates to a kind of cultural method of planktonic microalgae, be specifically related to a kind of method of genus bacillus regulation and control planktonic microalgae mixed culture.
Background technology
Little algae structure of community is stable not only relevant with the biomass of little algae in the water body environment, also closely related with its community diversity index, cultivate at present to little algae regulation and control or for the artificial orientation of planktonic microalgae, spininess is carried out single kind or two kinds of good little algaes, in the water body environment of complexity, cause indivedual little algae kind excessive multiplications of a certain stage easily, little algae community diversity index in the water body is on the low side relatively, be unfavorable for the stable of little algae group in the water surrounding, thus cause mid-early stage algae difficultly mutually cultivate, the situation that easily suddenlys change mid-term, the later stage is vigorous excessively.
Be the little algae of advantage through investigation Nannochloropsis oceanica, latent algae and green in the shrimp culture pond algae that quivers, wherein the above two are considered to good little algae more, can effectively regulate and control and optimize water body environment, and the algae that quivers is considered to harmful little algaes of culture fishery then more as blue-green algae.The green algae that quivers mostly is common kind in various aquaculture patterns in fact, even existence is also arranged in good water body environment and little algae structure of community more, under multiple little algae coexistence growing state, there is the green of the some amount algae that quivers, can keep the balance of little algae kind and ecological functions in the water body, but key must be strict controlled in quantity certain level, to avoid the dominance level excessive, hinder even suppress the growth of other good little algaes.
Have extremely close getting in touch between the microorganism in the water body environment and little algae.Some bacterium can be constantly to some organic substances of cell exocrine, and as the outer glycopeptide material of born of the same parents, the bacterium that has can also secrete the biostearin material, promotes some micro algae growth; The bacterium secretion that also has can be dissolved enzyme material or other active substances of frustule, and the micro algae growth of some kind is suppressed.Based on the specificity that is had between bacterium and algae, can from particular environment, screen relevant bacterial classification, and with the syntrophism system of the multiple little algae of regulation and control, make in the system the little algae of different sorts to grow than ratio of greater inequality example and quantity.Bacillus licheniformis all has stronger product extracellular enzyme activity as probiotics, can promote the matter and energy circulation in the water surrounding, and can also produce multiple antimicrobial substance, suppresses the growth of pathogeny bacterium.Since have nontoxic, have no side effect, characteristics such as pollution-free, noresidue, it is considered to beneficial microorganism safely and effectively, and widespread use and industries such as agricultural, medicine, food, feed processing.
Therefore, when regulating and control aquaculture and use little algae initial-stage culture, the specificity relation that is had between ecosystem characterization, bacterium and algae according to the little algae of different sorts, basis from the bacteria-algae synergism regulation and control, in little algae mixed culture system, introduce beneficial microorganism, improve little algae total number, stablize the different microalgae quantitative proportion, thereby be many algaes liquid of in middle-size and small-size prawn culturing system, introducing efficient stable, promote and optimization production initial stage water surrounding in little algal structure, and the present problem that will solve be badly in need of just about this research.
Summary of the invention
The object of the present invention is to provide a kind of method of genus bacillus regulation and control planktonic microalgae mixed culture, this method is grown each little algae in mixed culture balanced, good stability, avoid the species diversity of algae phase in the mixed culture system single, avoid the quantity of certain little algae extreme advantage or inferior position to occur.
The method of genus bacillus regulation and control planktonic microalgae mixed culture provided by the invention, choosing dominance, to place nitrogen concentration greater than little algae of 0.15 be that 12～16mg/L, phosphorus concentration are that 2～6mg/L, concentration of iron are that the nutrient solution of 0.10～0.20mg/L carries out mixed culture, in the mixed culture process, adopt Bacillus licheniformis to regulate and control, culture condition is: 26～28 ℃ of temperature, salinity 12～18 ‰, pH7.5～8.0, intensity of illumination 4000～5000Lx, Light To Dark Ratio 12h: 12h～14h: 10h.
In nutrient solution:
Nitrogenous source is one or more in ammonium nitrate, ammonium chloride, ammonium sulfate, SODIUMNITRATE, saltpetre or the urea.
The phosphorus source is one or more in potassium primary phosphate, SODIUM PHOSPHATE, MONOBASIC or the calcium superphosphate.
Source of iron is an ironic citrate.
Described little algae is the mixing of Nannochloropsis oceanica, latent algae and the green algae that quivers.
The input ratio of described Nannochloropsis oceanica, latent algae and the green algae that quivers is 0.7～1.3: 0.7～1.3: 0.7～1.3, and the initial input density of each little algae is 104～10
5Cells/mL.
The initial input density of described genus bacillus is 10
4～10
5CFU/mL.
At the aquaculture water water colour is emerald green, pea green or green and brown look, and aquaculture water planktonic microalgae biodiversity index is greater than in 1.60 the breed pond, screens excellently to go out the gesture degree and carry out mixed culture greater than little algae kind of 0.15.
The invention has the beneficial effects as follows:
(1) filters out the little algae of multiple advantage and carried out mixed culture, guaranteed the diversity of little algae algae phase, and, regulated and control the quantitative proportion of various little algaes directly or indirectly by the interpolation of Bacillus licheniformis in culture systems, make little algae reach the effect of balanced growth;
(2) adopting relevant function method aspect the dependency of genus bacillus and the growth of various microalgae cell, and adopted the path analysis method to estimate the influence of genus bacillus to various little algaes, pass through actual verification, draw the relevant regulation measure that the algae phase stability is cultivated, guaranteed the stability of various micro algae growths;
(3) cultivate with the symbiosis of the little algae of Bacillus licheniformis balance regulation different sorts, and then optimize the technology mode that algae is regulated and control mutually, this can be used in little algae ecotope of optimizing the prawn culturing pond, avoid in the breeding process algae phase species diversity that causes because of the little algae excessive multiplication of certain single kind in the pond or extinction single, water surrounding changes violent, situations such as aquaculture water environmental quality deterioration possess actual operation;
(4) the inventive method can be used as a kind of method that little algae initial-stage culture is used in aquaculture of regulating and control, its effect is the basis from the bacteria-algae synergism regulation and control, in little algae mixed culture system, improve little algae total number, stablize the different microalgae quantitative proportion, thereby carry out the initial stage algae in the middle-size and small-size prawn culturing system and make up mutually with optimization tachnical storage is provided for little algae of high-efficient culture is introduced.
Embodiment
Following examples only are used to set forth the present invention, and protection scope of the present invention is not only to be confined to following examples.The those of ordinary skill of described technical field all can be realized purpose of the present invention according to above content disclosed by the invention and scope that each parameter is got.
Embodiment 1
Select Nannochloropsis oceanica (Nannochloropsis oculata), latent algae (Cryptomonas erosa) and the green algae (Oscillatoria chlorine) that quivers to carry out mixed culture, use Bacillus licheniformis (Bacilluslicheniformis) in the culturing process and regulate and control, make the various little algae steady growth by a certain percentage in the system.
Nutrient solution is inorganic nutrient solution for total man worker disposes, and old extra large water boil cooling back adds NaNO
3, KH
2PO
4And ironic citrate, make the concentration of N, P and Fe reach 15mg/L, 2mg/L and 0.12mg/L respectively.The concentration of every nitrogen of 3d mensuration and phosphorus takes the circumstances into consideration to add NaNO in the culturing process
3And KH
2PO
4, make the concentration of total nitrogen and phosphorus reach initial level.
The correlated condition unanimity of each cultivation group in the culturing process, with constant temperature circulator water-bath temperature control, temperature is 26 ℃, pH8.2, intensity of illumination 5000Lx, Light To Dark Ratio are 14h: 10h, the salinity of nutritive medium is 16 ‰.
Algae mutually in the input ratio of each Nannochloropsis oceanica, latent algae and the green algae that quivers be 0.8: 1.1: 1.0, the initial input density of each little algae is 8 * 10
4Cells/mL; The initial input cell density of bacillus cell is 3 * 10
4CFU/mL; Culture vessel is 1M
3White transparent glass steel drum, cover bung with white light-passing plastic film, inflate cultivation, the process that spreads cultivation is cultivated algae liquid according to above-mentioned regulation measure and is managed, and in time adds the nutrient solution of corresponding proportion.
Present method practice shows in the mixed culture system, the cultivation initial stage 2～4d genus bacillus Nannochloropsis oceanica is had promoter action, but when Nannochloropsis oceanica density greater than 10
6Genus bacillus then suppresses the growth of Nannochloropsis oceanica behind the ind/mL; Latent algae all can be subjected to the growth promoting function of genus bacillus in whole mixed culture process, after cultivating 6 days, latent frustule number has improved more than 55% than control group; Though genus bacillus is little at the direct effect degree of the algae that quivers, by the growth of Nannochloropsis oceanica and latent algae in promotion or the regulation and control mixed culture system, thereby the quantity of little algae is controlled in the mesotrophic consumption of acceleration system indirectly.
The total cell count of little algae of adding genus bacillus after cultivating 14d in the system is 10.6 * 10
6Ind/mL, and the cell count of Nannochloropsis oceanica is 4.8 * 10
6Ind/mL more do not add genus bacillus reduction by 40.6%, latent algae and the green algae that quivers then is respectively 3.3 * 10
6Ind/mL and 2.5 * 10
6Ind/mL, that does not more add genus bacillus has raise 21.0% and 2.5% respectively.As seen, culturing process promotes or regulates and control the growth of various algaes in the mixed culture system by the interpolation that utilizes Bacillus licheniformis, or thereby the quantity of little algae is controlled in the mesotrophic consumption of acceleration system indirectly, effectively regulate and control the quantitative proportion of various little algaes, reach the effect of balanced growth, avoid the quantity of certain little algae extreme advantage or inferior position to occur.
Embodiment 2
Select Nannochloropsis oceanica (Nannochloropsis oculata), latent algae (Cryptomonas erosa) and the green algae (Oscillatoria chlorine) that quivers to carry out mixed culture, use Bacillus licheniformis (Bacilluslicheniformis) in the culturing process and regulate and control, make the various little algae steady growth by a certain percentage in the system.
Nutrient solution is inorganic nutrient solution for total man worker disposes, and old extra large water boil cooling back adds NaNO
3, KH
2PO
4And ironic citrate, make the concentration of N, P and Fe reach 16mg/L, 3mg/L and 0.14mg/L respectively.The concentration of every nitrogen of 3d mensuration and phosphorus takes the circumstances into consideration to add NaNO in the culturing process
3And KH
2PO
4, make the concentration of total nitrogen and phosphorus reach initial level.
The correlated condition unanimity of each cultivation group in the culturing process, with constant temperature circulator water-bath temperature control, temperature is 28 ℃, pH7.8, intensity of illumination 4800Lx, Light To Dark Ratio are 14h: 10h, the salinity of nutritive medium is 18 ‰.
Algae mutually in the input ratio of each Nannochloropsis oceanica, latent algae and the green algae that quivers be 0.8: 1.1: 1.2, the initial input density of each little algae is 10
4Cells/mL; The initial input cell density of bacillus cell is 10
4CFU/mL.Culture vessel is 1M
3White transparent glass steel drum, cover bung with white light-passing plastic film, inflate cultivation; The process that spreads cultivation is cultivated algae liquid according to above-mentioned regulation measure and is managed, and in time adds the nutrient solution of corresponding proportion.
Present method practice shows in the mixed culture system, the cultivation initial stage 1～4d genus bacillus Nannochloropsis oceanica is had promoter action, but when Nannochloropsis oceanica density greater than 6.0 * 10
5Genus bacillus then suppresses the growth of Nannochloropsis oceanica behind the ind/mL; Latent algae all can be subjected to the growth promoting function of genus bacillus in whole mixed culture process, after cultivating 5 days, latent frustule number has improved more than 50% than control group; Though genus bacillus is little at the direct effect degree of the algae that quivers, by the growth of Nannochloropsis oceanica and latent algae in promotion or the regulation and control mixed culture system, thereby the quantity of little algae is controlled in the mesotrophic consumption of acceleration system indirectly.
The total cell count of little algae of adding genus bacillus after cultivating 14d in the system is 8.0 * 10
6Ind/mL, and the cell count of Nannochloropsis oceanica is 3.2 * 10
6Ind/mL more do not add genus bacillus reduction by 38.0%, latent algae and the green algae that quivers then is respectively 2.8 * 10
6Ind/mL and 2.0 * 10
6Ind/mL, that does not more add genus bacillus has raise 18% and 2.5% respectively.As seen, culturing process promotes or regulates and control the growth of various algaes in the mixed culture system by the interpolation that utilizes Bacillus licheniformis, or thereby the quantity of little algae is controlled in the mesotrophic consumption of acceleration system indirectly, effectively regulate and control the quantitative proportion of various little algaes, reach the effect of balanced growth, avoid the quantity of certain little algae extreme advantage or inferior position to occur.
Embodiment 3
Selecting the pond water colour is emerald green, pea green or green and brown look, aquaculture water planktonic microalgae biodiversity index is greater than 1.60, the prawn seawater aquaculturing pond that production performance is good carries out the all-the-way tracking analysis, the screening dominance is greater than the Nannochloropsis oceanica more than 0.15 (Nannochloropsis oculata), latent algae (Cryptomonas erosa) and the green algae (Oscillatoria chlorine) that quivers carries out mixed culture, use Bacillus licheniformis (Bacillus licheniformis) in the culturing process and regulate and control, make the various little algae steady growth by a certain percentage in the system.Adopt the simple correlation analytical method to analyze the dependency of genus bacillus and the growth of various microalgae cell in the culturing process; Estimate the influence of genus bacillus with the path analysis method to various little algaes.
Nutrient solution is inorganic nutrient solution for total man worker disposes, and old extra large water boil cooling back adds KNO
3, Ca (H
2PO
4)
2And ironic citrate, make the concentration of N, P and Fe reach 14mg/L, 2mg/L and 0.14mg/L respectively.The concentration of every nitrogen of 3d mensuration and phosphorus takes the circumstances into consideration to add KNO in the culturing process
3And Ca (H
2PO
4)
2, make the concentration of total nitrogen and phosphorus reach initial level.
The correlated condition unanimity of each cultivation group in the culturing process, with constant temperature circulator water-bath temperature control, temperature is 26～28 ℃, pH7.8, intensity of illumination 4000Lx, Light To Dark Ratio are 12h: 12h, the salinity of nutritive medium is 15 ‰.
Algae mutually in the input ratio of each Nannochloropsis oceanica, latent algae and the green algae that quivers be 1.0: 1.0: 1.0, the initial input density of each little algae is 7 * 10
4Cells/mL; The initial input cell density of bacillus cell is 5 * 10
4CFU/mL.Culture vessel is 1M
3White transparent glass steel drum, cover bung with white light-passing plastic film, inflate cultivation.
The process that spreads cultivation is cultivated algae liquid according to above-mentioned regulation measure and is managed, and in time adds the nutrient solution of corresponding proportion.Culturing process promotes or regulates and control the growth of various algaes in the mixed culture system by the interpolation that utilizes Bacillus licheniformis, or thereby the quantity of little algae is controlled in the mesotrophic consumption of acceleration system indirectly, effectively regulate and control the quantitative proportion of various little algaes, reach the effect of balanced growth, avoid the quantity of certain little algae extreme advantage or inferior position to occur.
Present method practice shows in the mixed culture system, the cultivation initial stage 1～4d genus bacillus Nannochloropsis oceanica is had promoter action, but when Nannochloropsis oceanica density greater than 6.5 * 10
5Genus bacillus then suppresses the growth of Nannochloropsis oceanica behind the ind/mL; Latent algae all can be subjected to the growth promoting function of genus bacillus in whole mixed culture process, after cultivating 6 days, latent frustule number has improved 51.6% than control group; Though genus bacillus is little at the direct effect degree of the algae that quivers, by the growth of Nannochloropsis oceanica and latent algae in promotion or the regulation and control mixed culture system, thereby the quantity of little algae is controlled in the mesotrophic consumption of acceleration system indirectly.
The total cell count of little algae of adding genus bacillus after cultivating 16d in the system is 8.5 * 10
6Ind/mL, and the cell count of Nannochloropsis oceanica is 2.9 * 10
6Ind/mL more do not add genus bacillus reduction by 43.5%, latent algae and the green algae that quivers then is respectively 2.9 * 10
6Ind/mL and 2.7 * 10
6Ind/mL, that does not more add genus bacillus has raise 19.2% and 3.1% respectively.
In general, by in culture system, adding the growth that genus bacillus both can effectively have been improved the little algae of individual species on the one hand, can effectively regulate and control the quantitative proportion of various little algaes again, reach the effect of balanced growth, avoid the quantity of certain little algae extreme advantage or inferior position to occur.
Claims (8)
1, a kind of method of genus bacillus regulation and control planktonic microalgae mixed culture, it is characterized in that, choosing dominance, to place nitrogen concentration greater than little algae of 0.15 be that 12～16mg/L, phosphorus concentration are that 2～6mg/L, concentration of iron are that the nutrient solution of 0.10～0.20mg/L carries out mixed culture, in the mixed culture process, adopt Bacillus licheniformis to regulate and control, culture condition is: 26～28 ℃ of temperature, salinity 12～18 ‰, pH7.5～8.0, intensity of illumination 4000～5000Lx, Light To Dark Ratio 12h: 12h～14h: 10h.
2, the method for genus bacillus regulation and control planktonic microalgae mixed culture according to claim 1 is characterized in that nitrogenous source is one or more in ammonium nitrate, ammonium chloride, ammonium sulfate, SODIUMNITRATE, saltpetre or the urea in the nutrient solution.
3, the method for genus bacillus regulation and control planktonic microalgae mixed culture according to claim 1 is characterized in that the phosphorus source is one or more in potassium primary phosphate, SODIUM PHOSPHATE, MONOBASIC or the calcium superphosphate in the nutrient solution.
4, the method for genus bacillus regulation and control planktonic microalgae mixed culture according to claim 1 is characterized in that source of iron is an ironic citrate in the nutrient solution.
5, the method for genus bacillus regulation and control planktonic microalgae mixed culture according to claim 1 is characterized in that, described little algae is the mixing of Nannochloropsis oceanica, latent algae and the green algae that quivers.
6, the method for genus bacillus regulation and control planktonic microalgae mixed culture according to claim 5, it is characterized in that, the input ratio of described Nannochloropsis oceanica, latent algae and the green algae that quivers is 0.7～1.3: 0.7～1.3: 0.7～1.3, and the initial input density of each little algae is 10
4～10
5Cells/mL.
7, the method for genus bacillus regulation and control planktonic microalgae mixed culture according to claim 1 is characterized in that the initial input density of described genus bacillus is 10
4～10
5CFU/mL.
8, the method for genus bacillus regulation and control planktonic microalgae mixed culture according to claim 1, it is characterized in that, at the aquaculture water water colour is emerald green, pea green or green and brown look, aquaculture water planktonic microalgae biodiversity index filters out dominance and carries out mixed culture greater than little algae kind of 0.15 greater than in 1.60 the breed pond.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103773692A (en) * | 2012-10-23 | 2014-05-07 | 中国石油化工股份有限公司 | Enclosed culture method of microalgae |
| CN103773695A (en) * | 2012-10-23 | 2014-05-07 | 中国石油化工股份有限公司 | Open-type rapid culture method of microalgae |
| CN103773691A (en) * | 2012-10-23 | 2014-05-07 | 中国石油化工股份有限公司 | Enclosed rapid culture method of microalgae |
| CN104630068A (en) * | 2015-02-13 | 2015-05-20 | 新奥科技发展有限公司 | Culturing method of microalgae and culture medium thereof |
| CN106148197A (en) * | 2016-08-31 | 2016-11-23 | 天津海友佳音生物科技股份有限公司 | A kind of method of bacillus cereus cultivating microalgae |
| CN106566785A (en) * | 2016-10-31 | 2017-04-19 | 徐州工程学院 | Probiotic, inoculant as well as preparation method and applications of inoculant |
| CN107522281A (en) * | 2017-10-19 | 2017-12-29 | 张永华 | A kind of preparation method for the algicide administered for water-surface blue-green algae |
| CN108085283A (en) * | 2018-01-19 | 2018-05-29 | 何红娣 | A kind of helotism high density Algaculture method |
| CN109402025A (en) * | 2018-12-12 | 2019-03-01 | 河西学院 | A kind of both culturing microalgae prevention and cure of pollution method |
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| CN103773692B (en) * | 2012-10-23 | 2016-11-09 | 中国石油化工股份有限公司 | A kind of method of closed cultivation microalgae |
| CN103773695A (en) * | 2012-10-23 | 2014-05-07 | 中国石油化工股份有限公司 | Open-type rapid culture method of microalgae |
| CN103773691A (en) * | 2012-10-23 | 2014-05-07 | 中国石油化工股份有限公司 | Enclosed rapid culture method of microalgae |
| CN103773692A (en) * | 2012-10-23 | 2014-05-07 | 中国石油化工股份有限公司 | Enclosed culture method of microalgae |
| CN103773695B (en) * | 2012-10-23 | 2016-08-17 | 中国石油化工股份有限公司 | A kind of method of open type fast culture microalgae |
| CN103773691B (en) * | 2012-10-23 | 2016-10-26 | 中国石油化工股份有限公司 | A kind of method of closed fast culture microalgae |
| CN104630068A (en) * | 2015-02-13 | 2015-05-20 | 新奥科技发展有限公司 | Culturing method of microalgae and culture medium thereof |
| CN106148197A (en) * | 2016-08-31 | 2016-11-23 | 天津海友佳音生物科技股份有限公司 | A kind of method of bacillus cereus cultivating microalgae |
| CN106566785A (en) * | 2016-10-31 | 2017-04-19 | 徐州工程学院 | Probiotic, inoculant as well as preparation method and applications of inoculant |
| US10927339B2 (en) | 2017-03-17 | 2021-02-23 | Industrial Technology Research Institute | Mutant of Bacillus thuringiensis and application thereof |
| CN107522281A (en) * | 2017-10-19 | 2017-12-29 | 张永华 | A kind of preparation method for the algicide administered for water-surface blue-green algae |
| CN108085283A (en) * | 2018-01-19 | 2018-05-29 | 何红娣 | A kind of helotism high density Algaculture method |
| CN108085283B (en) * | 2018-01-19 | 2019-12-06 | 杭州渔森农业技术开发有限公司 | method for culturing high-density algae through symbiosis of bacteria and algae |
| CN109402025A (en) * | 2018-12-12 | 2019-03-01 | 河西学院 | A kind of both culturing microalgae prevention and cure of pollution method |
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