CN105861395A - Composite bacterial agent DH-1 capable of effectively inhibiting water-bloom microcystis aeruginosa and application thereof - Google Patents
Composite bacterial agent DH-1 capable of effectively inhibiting water-bloom microcystis aeruginosa and application thereof Download PDFInfo
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- CN105861395A CN105861395A CN201610421245.0A CN201610421245A CN105861395A CN 105861395 A CN105861395 A CN 105861395A CN 201610421245 A CN201610421245 A CN 201610421245A CN 105861395 A CN105861395 A CN 105861395A
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Abstract
The invention discloses a composite bacterial agent DH-1 capable of effectively inhibiting water-bloom microcystis aeruginosa and an application thereof. The composite bacterial agent DH-1 comprises Pseudomona bacteria, Shinella bacteria, Thermomonas bacteria, Devosia bacteria and Alishewanella bacteria. According to the composite bacterial agent DH-1 and the application thereof, the specific function microorganism-composite bacterial agent DH-1 is put into a water body, the hydrophilous micro-ecology adjusting effect and the phycomycetes acting effect of microorganisms are played, and therefore the microorganism algae removing method for inhibiting (removing) microcystis aeruginosa is achieved; the composite bacterial agent DH-1 has the advantages that cost is low, security is high, and the ecological environment of the water body is not damaged, and the composite bacterial agent DH-1 is of important-practical significance in treating a microcystis-aeruginosa water-bloom water body.
Description
Technical field
The invention belongs to microbe application field, be specifically related to the composite bacteria agent capable DH-1 of a kind of effective suppression wawter bloom microcystic aeruginosa
And application.
Background technology:
Along with urbanization and the development of industrial and agricultural production, in a large number rich in industrial wastewater, the life dirt of the nutrient substance such as nitrogen (N), phosphorus (P)
Water flows into the slow flow water bodies such as lake, river, makes water Middle nutrition material be enriched with rapidly, and in water, algae fulminant increases, blue-green alga bloom
Frequently occur, cause water body serious eutrophication, bring adverse effect to aspects such as Water quality, water safety and landscape environments.
Microcystic aeruginosa (Microcystis aeruginosa) is the most common algae kind causing wawter bloom, the main algae of Ye Shi China blue-green alga bloom
Kind, its special physiological ecological feature makes it easily become advantage algae kind in eutrophication water and cause breaking out of blue-green alga bloom.
Microcystic aeruginosa can discharge hepatotoxin in cell growth and death process, and this toxin is a kind of strong carcinogenic promoting agent, to aquatile
Very harmful, the health of the indirect hazard mankind, therefore, administer blue-green alga bloom and pollute the problem having become urgent solution.
The administering method of blue-green alga bloom mainly has Physical and chemical method at present, and these methods exist certain defect, such as Physical
Cost is high, need a large amount of manpower and materials and disposal ability limited.Chemical method easily causes secondary pollution, selects toxicity difference or only
Adapt to little area waters.Therefore, actively find blue-green alga bloom prevention and controls novel, efficient, environmental protection and have highly important
Meaning.
Summary of the invention:
It is an object of the invention to provide a kind of composite bacteria agent capable DH-1 that can effectively suppress wawter bloom microcystic aeruginosa and application thereof.
The composite bacteria agent capable DH-1 that can effectively suppress wawter bloom microcystic aeruginosa of the present invention, it is characterised in that containing Rhodopseudomonas
The antibacterial of (Pseudomona sp.), Shinella sp. belong to the antibacterial of (Shinella sp.), hot zygosaccharomyces (Thermomonas sp.)
The antibacterial of uncommon Bordetella (the Alishewanella sp.) that go out of antibacterial, the antibacterial of De Wosi Bordetella (Devosia sp.) and abnormal type.
Preferably, in the described composite bacteria agent capable DH-1 that can effectively suppress wawter bloom microcystic aeruginosa, the antibacterial of Rhodopseudomonas, Shen
The antibacterial of family name's Bacillus, the antibacterial of hot zygosaccharomyces, the antibacterial of De Wosi Bordetella and abnormal type wish the bacterial number ratio of Bordetella of going out
For 160:27:8:3:1.
Preferably, the antibacterial of described Rhodopseudomonas is preferably pseudomonas mendocina (Pseudomonas mendocina)
ACCC01078 or pseudomonas (Pseudomonas sp.) FIM-20151013001；The antibacterial that described Shinella sp. belongs to is
Shinella sp. (Shinella sp.) ACCC19821 or Shinella sp. (Shinella sp.) FIM-20151013002；Described heat is single
The antibacterial of born of the same parents Pseudomonas is hot Zymomonas mobilis (Thermomonas sp.) FIM-20151013003；The antibacterial of described De Wosi Bordetella is
Moral Butterworth Salmonella (Devosia honganense) ACCC19737 or De Wosi Salmonella (Devosia sp.) FIM-20151013004,
Described abnormal type is wished the antibacterial of Bordetella of going out and is preferably abnormal type uncommon go out Salmonella (Alishewanella fetalis) ACCC10045 or abnormal type
Uncommon Salmonella (Alishewanella sp.) FIM-20151013005 that goes out.
Second object of the present invention is to provide the preparation method of the composite bacteria agent capable DH-1 that can effectively suppress wawter bloom microcystic aeruginosa,
It is characterized in that, by the antibacterial of Rhodopseudomonas, the antibacterial of Shinella sp. genus, the antibacterial of hot zygosaccharomyces, De Wosi Bordetella
Antibacterial and abnormal type wish Bordetella of going out antibacterial bacterium solution mixing after, go bacteria-removing liquid to stay thalline, thalline to be composite bacteria agent capable DH-1.
Preferably, it is respectively by the antibacterial of Rhodopseudomonas, the antibacterial of Shinella sp. genus, the antibacterial of hot zygosaccharomyces, De Wosishi
The antibacterial of Pseudomonas and abnormal type are wished the antibacterial of Bordetella of going out and are fermented respectively, and being then diluted to cell concentration is 1 × 108Individual/ml, then
By 1 × 108The antibacterial of the Rhodopseudomonas of individual/ml, the antibacterial of Shinella sp. genus, the antibacterial of hot zygosaccharomyces, De Wosi Bordetella
Antibacterial and abnormal type wish the antibacterial of Bordetella of going out according to volume ratio 160:27:8:3:1 mix homogeneously, be then centrifuged for removing supernatant,
Thalline, thalline is stayed to be composite bacteria agent capable DH-1.
Third object of the present invention is to provide the composite bacteria agent capable DH-1 that can effectively suppress wawter bloom microcystic aeruginosa at suppression wawter bloom copper
Application in green Microcystis aeruginosa.
Wawter bloom microcystic aeruginosa function bacterium, according to microbial ecology principle and method, is rationally compounded, it is thus achieved that right by the present invention
Microcystic aeruginosa has the composite bacteria agent capable DH-1 of good suppression (removal) effect, has the value of popularization and application.
The present invention, by putting into specific functional microorganism-composite bacteria agent capable DH-1 in water body, plays the regulation of microorganism aquatic Tiny ecosystem
The effect of " phycomycete interaction ", thus reach to suppress the microorganism of (removal) microcystic aeruginosa to remove algae method, there is low cost, peace
Quan Xinggao and do not destroy the characteristic of water ecological setting, has important practical significance to administering microcystic aeruginosa wawter bloom water body.
Detailed description of the invention:
Following example are to further illustrate the present invention rather than limitation of the present invention.
Pseudomonas (Pseudomonas sp.) FIM-20151013001, the Shinella sp. (Shinella used in following example
Sp.) FIM-20151013002, hot Zymomonas mobilis (Thermomonas sp.) FIM-20151013003, De Wosi Salmonella (Devosia sp.)
FIM-20151013004 and abnormal type are wished and are gone out Salmonella (Alishewanella sp.) FIM-20151013005 both from the micro-life in Fujian Province
Thing institute, it is preserved in Fujian Microorganism Inst., and the bacterial strain being preserved in Fujian Microorganism Inst. is for sale,
These bacterial strains are also for sale, and therefore before the applying date, of the prior art anyone can buy these bacterial strains.
And above-mentioned bacterial strains had been proposed on webpage before the applying date
(http://www.fjim.org.cn/kytd/ktz/getNews.jsp？Newsid=220), this bacterial strain the applicant also hold, and ensures certainly
The applying date provides to the public in rising 20 years.
Embodiment 1:
1, slant culture
By pseudomonas (Pseudomonas sp.) FIM-20151013001, Shinella sp. (Shinella sp.)
FIM-20151013002, hot Zymomonas mobilis (Thermomonas sp.) FIM-20151013003, De Wosi Salmonella (Devosia sp.)
FIM-20151013004 and abnormal type uncommon go out Salmonella (Alishewanella sp.) FIM-20151013005 and Pseudomonas aeruginosa
(Pseudomonas aeruginosa), pseudomonas putida (Pseudomonas putida) and pseudomonas mendocina
(Pseudomonas mendocina) is seeded on nutrient agar, cultivates 2～3d in 28 DEG C of incubators.
Nutrient agar formula: peptone 10g, Carnis Bovis seu Bubali cream 3g, sodium chloride 5g, agar 15～20g, distilled water 1000mL,
After mix homogeneously, 121 DEG C of autoclaving 15min.
2, fermentation culture
The thalline provoking above-mentioned bacterial strains on a small amount of slant medium with inoculating loop respectively accesses in fermentation medium, in 28 DEG C, 150
Shaken cultivation 2～3d on the shaking table of r/min.With fermentation medium, bacterium solution is diluted so that it is concentration is 1 × 108～109Individual/ml.
Fermentative medium formula (g/L): tryptone 10.0, yeast powder 5.0, sodium chloride 10.0, surplus is water, and pH is natural, by upper
After stating composition mix homogeneously, sterilizing is standby.
3, the preparation of composite bacteria agent capable
By 1 × 108The pseudomonas bacterium solution of individual/ml, 1 × 108The Shinella sp. bacterium solution of individual/ml, 1 × 108The hot Zymomonas mobilis bacterium solution of individual/ml,
1×108The De Wosi Salmonella bacterium solution of individual/ml and 1 × 108The uncommon Salmonella bacterium solution of going out of the abnormal type of individual/ml, according to volume ratio 160:27:8:3:
The volume ratio mix homogeneously of 1,8000r/min is centrifuged 5min, abandons supernatant and obtains thalline, is composite bacteria agent capable, by this compound bacteria
Agent named composite bacteria agent capable DH-1.
The inhibitory action of microcystic aeruginosa is tested by embodiment 2 composite bacteria agent capable DH-1 and Rhodopseudomonas strain
Gathering Foochow certain blue-green alga bloom river permanently happy, these blue-green alga bloom sociales are verdigris color Microcystis aeruginosa.River is in advance through 8 layers of gauze
Filter and remove large particulate matter.Measure 2L river, join in 5L beaker, be simultaneously introduced 1L BG-11 culture medium, stir evenly mixed
All.Using the group without bacterium as blank group (CK), to add the composite bacteria agent capable DH-1 of embodiment 1, Aerugo vacation respectively
Zymomonas mobilis, pseudomonas putida, pseudomonas mendocina, Shinella sp. (Shinella sp.) FIM-20151013002, heat list
Born of the same parents bacterium (Thermomonas sp.) FIM-20151013003, De Wosi Salmonella (Devosia sp.) FIM-20151013004 and abnormal type
The group of uncommon Salmonella (Alishewanella sp.) FIM-20151013005 that goes out is experimental group, is all provided with three Duplicate Samples, Pseudomonas aeruginosa,
Pseudomonas putida, pseudomonas mendocina, Shinella sp. (Shinella sp.) FIM-20151013002, hot Zymomonas mobilis
(Thermomonas sp.) FIM-20151013003, De Wosi Salmonella (Devosia sp.) FIM-20151013004 and abnormal type are wished and are gone out
The thalline addition of Salmonella (Alishewanella sp.) FIM-20151013005 is identical with the addition of composite bacteria agent capable DH-1.Due to
Chlorophyll a (chlorophyll-a, chl-a) plays absorption and transmission light as the important extinction pigment of alga cells in photosynthesis
Can pivotal role, the growth to algae plays material impact, is one of the important indicator of vegetation growth state, so with chlorophyll a
Content as the evaluation index of algae content.Use acetone extraction spectrophotography (People's Republic of China's water conservancy industry standard
(SL 88-2012)) measure Chlorophyll-a Content.
Chlorophyll-a Content in the initial water body of measured in advance.Experimental group adds composite bacteria agent capable by the percent by volume of 1 ‰ respectively
DH-1, Pseudomonas aeruginosa, pseudomonas putida, pseudomonas mendocina, Shinella sp. (Shinella sp.)
FIM-20151013002, hot Zymomonas mobilis (Thermomonas sp.) FIM-20151013003, De Wosi Salmonella (Devosia sp.)
FIM-20151013004 and uncommon Salmonella (Alishewanella sp.) FIM-20151013005 that goes out of abnormal type, bottom is provided with aeration head, adopts
With intermittent aerating, i.e. exposing 2h, stop 2h, experiment is carried out in illumination box, and condition of culture is (25 ± 1) DEG C, Light To Dark Ratio (light
According to time and the ratio without light application time) it is 12:12, intensity of illumination is 2000lux.Timing is observed.Measure in water body after cultivating 8d
Chlorophyll-a Content.
Test result indicate that (table 1), after cultivating 8d, blank group (CK) and addition composite bacteria agent capable DH-1, Aerugo vacation
Zymomonas mobilis, pseudomonas putida, pseudomonas mendocina, Shinella sp. (Shinella sp.) FIM-20151013002, heat list
Born of the same parents bacterium (Thermomonas sp.) FIM-20151013003, De Wosi Salmonella (Devosia sp.) FIM-20151013004 and abnormal type
Chlorophyll a average content in the process group water body of uncommon Salmonella (Alishewanella sp.) FIM-20151013005 that goes out is respectively
1.349 μ g/L, 0.21 μ g/L, 0.60 μ g/L, 0.69 μ g/L, 0.63 μ g/L, 1.01 μ g/L, 0.98 μ g/L, 1.07 μ g/L and
0.95μg/L.Chlorophyll a compared with the Chlorophyll-a Content 0.29 μ g/L in initial water body, in blank group (CK) water body
Content has significantly rising, and the microcystic aeruginosa illustrated in blank group after 8d cultivates mushrooms out breeding, and content is obvious
Improve.Compared with blank group (CK), with the addition of Rhodopseudomonas Pseudomonas aeruginosa, pseudomonas putida, Mendoza
Chlorophyll-a Content in the process group water body of pseudomonas has rising by a small margin, and Rhodopseudomonas Pseudomonas aeruginosa, evil are described
Smelly pseudomonas and pseudomonas mendocina have certain inhibition to microcystic aeruginosa, and add Shinella sp. (Shinella
Sp.) FIM-20151013002, hot Zymomonas mobilis (Thermomonas sp.) FIM-20151013003, De Wosi Salmonella (Devosia sp.)
Leaf in the process group water body of FIM-20151013004 and uncommon Salmonella (Alishewanella sp.) FIM-20151013005 that goes out of abnormal type
Verdazulene a content has certain rising, and Shinella sp. (Shinella sp.) FIM-20151013002, hot Zymomonas mobilis are described
(Thermomonas sp.) FIM-20151013003, De Wosi Salmonella (Devosia sp.) FIM-20151013004 and abnormal type are wished and are gone out
Salmonella (Alishewanella sp.) FIM-20151013005 does not have significant inhibition to microcystic aeruginosa, and adds compound bacteria
The process group of agent DH-1 and blank group (CK) and interpolation Pseudomonas aeruginosa, pseudomonas putida and Mendoza vacation unit cell
The process group of bacterium is compared, and the Chlorophyll-a Content change in its water body is inconspicuous, illustrates that composite bacteria agent capable DH-1 is to wawter bloom Aerugo microcapsule
Algae has strong inhibitory action, and it is micro-that composite bacteria agent capable DH-1 has more preferably suppression wawter bloom Aerugo than single pseudomonas strain
The effect of capsule algae.
Table 1 experimental bacteria suppression result to microcystic aeruginosa
BG11 nutrient media components: NaNO31.5g, K2HPO40.04g, MgSO4·7H2O 0.075g, CaCl2·7H2O 0.036
G, Na2CO30.02g, citric acid 0.006g, ferric citrate 0.006g, trace element solution A5 1ml, ammonia benzyl penicillium sp
Element (final concentration) 50 μ g/mL, water is settled to 1000mL.Trace element solution A5 component: H3BO42.86g, MnCl2·4H2O
1.81g, ZnSO40.222g, Na2MoO40.39g, CuSO4·5H2O 0.079g, Co (NO3)2·6H2O 49.4g, water
It is settled to 1000ml.
The present invention can also belong to by other antibacterials that pseudomonas (Pseudomona) belongs to, Shinella sp. (Shinella) other are thin
Other antibacterials that other antibacterials that bacterium, hot Zymomonas mobilis (Thermomonas) belong to, De Wosi Salmonella (Devosia) belong to and abnormal type is uncommon goes out
Other antibacterials that Salmonella (Alishewanella) belongs to, as by the pseudomonas (Pseudomonas sp.) in embodiment 1
FIM-20151013001 replaces with pseudomonas mendocina (Pseudomonas mendocina) ACCC01078, Shinella sp.
(Shinella sp.) FIM-20151013002 replaces with Shinella sp. (Shinella sp.) ACCC19821, De Wosi Salmonella
(Devosia sp.) FIM-20151013004 replaces with De Wosi Salmonella (Devosia honganense) ACCC19737, and atypical wishing is gone out
Salmonella (Alishewanella sp.) FIM-20151013005 replaces with atypical uncommon Salmonella (Alishewanella fetalis) of going out
ACCC10045, then according to the method for embodiment 1 prepares composite bacteria agent capable DH-1 (2), then according to said method is carried out
The experiment of suppression wawter bloom microcystic aeruginosa, wawter bloom microcystic aeruginosa is had strongly by experimental result display composite bacteria agent capable DH-1 (2)
Inhibitory action, composite bacteria agent capable DH-1 (2) than single pseudomonas strain have more preferably suppression wawter bloom microcystic aeruginosa
Effect.Described pseudomonas mendocina (Pseudomonas mendocina) ACCC01078, Shinella sp. (Shinella sp.)
ACCC19821, De Wosi Salmonella (Devosia honganense) ACCC19737, the atypical uncommon Salmonella (Alishewanella that goes out
Fetalis) ACCC10045 is all preserved in Chinese agriculture Microbiological Culture Collection administrative center, can buy therefrom.
Claims (7)
1. the composite bacteria agent capable DH-1 that can effectively suppress wawter bloom microcystic aeruginosa, it is characterised in that containing Rhodopseudomonas
The antibacterial of (Pseudomona sp.), Shinella sp. belong to the antibacterial of (Shinella sp.), hot zygosaccharomyces (Thermomonas sp.)
The antibacterial of uncommon Bordetella (the Alishewanella sp.) that go out of antibacterial, the antibacterial of De Wosi Bordetella (Devosia sp.) and abnormal type.
The composite bacteria agent capable DH-1 that can effectively suppress wawter bloom microcystic aeruginosa the most according to claim 1, it is characterised in that described
The composite bacteria agent capable DH-1 that can effectively suppress wawter bloom microcystic aeruginosa in, the antibacterial of Rhodopseudomonas, Shinella sp. belong to thin
Bacterium, the antibacterial of hot zygosaccharomyces, the antibacterial of De Wosi Bordetella and abnormal type wish the bacterial number of Bordetella of going out than for 160:27:
8:3:1.
The composite bacteria agent capable DH-1 that can effectively suppress wawter bloom microcystic aeruginosa the most according to claim 1, it is characterised in that described
The antibacterial of Rhodopseudomonas be pseudomonas mendocina (Pseudomonas mendocina) ACCC01078 or pseudomonas
(Pseudomonas sp.)FIM-20151013001；The antibacterial that described Shinella sp. belongs to is Shinella sp. (Shinella sp.)
ACCC19821 or Shinella sp. (Shinella sp.) FIM-20151013002；The antibacterial of described hot zygosaccharomyces is heat
Zymomonas mobilis (Thermomonas sp.) FIM-20151013003；The antibacterial of described De Wosi Bordetella is De Wosi Salmonella
(Devosia honganense) ACCC19737 or De Wosi Salmonella (Devosia sp.) FIM-20151013004, described is another
It is the uncommon uncommon Salmonella of going out of Salmonella (Alishewanella fetalis) ACCC10045 or atypical of going out of abnormal type that class wishes the antibacterial of Bordetella of going out
(Alishewanella sp.)FIM-20151013005。
4. a preparation method of the composite bacteria agent capable DH-1 that can effectively suppress wawter bloom microcystic aeruginosa described in claim 1, its feature
Be, by the antibacterial of Rhodopseudomonas, Shinella sp. belong to antibacterial, the antibacterial of hot zygosaccharomyces, De Wosi Bordetella thin
After bacterium and abnormal type wish the bacterium solution mixing of the antibacterial of Bordetella of going out, bacteria-removing liquid is gone to stay thalline, thalline to be composite bacteria agent capable DH-1.
Preparation method the most according to claim 4, it is characterised in that be respectively the antibacterial of Rhodopseudomonas, Shinella sp. to be belonged to
Antibacterial, the antibacterial of hot zygosaccharomyces, the antibacterial of De Wosi Bordetella and abnormal type wish the antibacterial of Bordetella of going out and ferment respectively,
Being then diluted to cell concentration is 1 × 108Individual/ml, then by 1 × 108The antibacterial of the Rhodopseudomonas of individual/ml, Shinella sp. belong to
Antibacterial, the antibacterial of hot zygosaccharomyces, the antibacterial of De Wosi Bordetella and abnormal type wish the antibacterial of Bordetella of going out according to volume ratio
160:27:8:3:1 mix homogeneously, is then centrifuged for removing supernatant, stays thalline, thalline to be composite bacteria agent capable DH-1.
Preparation method the most according to claim 4, it is characterised in that the antibacterial of described Rhodopseudomonas is Mendoza vacation unit cell
Bacterium (Pseudomonas mendocina) ACCC01078 or pseudomonas (Pseudomonas sp.) FIM-20151013001；
The antibacterial that described Shinella sp. belongs to is Shinella sp. (Shinella sp.) ACCC19821 or Shinella sp. (Shinella sp.)
FIM-20151013002；The antibacterial of described hot zygosaccharomyces is hot Zymomonas mobilis (Thermomonas sp.)
FIM-20151013003；The antibacterial of described De Wosi Bordetella is De Wosi Salmonella (Devosia honganense)
ACCC19737 or De Wosi Salmonella (Devosia sp.) FIM-20151013004, described abnormal type wishes the antibacterial of Bordetella of going out
For the uncommon uncommon Salmonella (Alishewanella sp.) of going out of Salmonella (Alishewanella fetalis) ACCC10045 or atypical of going out of abnormal type
FIM-20151013005。
7. the composite bacteria agent capable DH-1 that can effectively suppress wawter bloom microcystic aeruginosa described in claim 1,2 or 3 is suppressing wawter bloom Aerugo
Application in Microcystis aeruginosa.
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| CN108753652A (en) * | 2018-06-14 | 2018-11-06 | 佛山市碧沃丰生物科技股份有限公司 | A method of degradation seat algae |
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| CN103131637A (en) * | 2013-02-25 | 2013-06-05 | 黑龙江八一农垦大学 | Bacteria compsite inoculum used in lignin decomposition |
| CN105176868A (en) * | 2015-09-08 | 2015-12-23 | 福建省微生物研究所 | Complex bactericide for efficiently purifying sewage anaerobic outlet water of pig farm and preparation method and application thereof |
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| CN102719383A (en) * | 2012-06-29 | 2012-10-10 | 武汉工业学院 | Bacillus amyloliquefaciens, inoculant and application thereof |
| CN103131637A (en) * | 2013-02-25 | 2013-06-05 | 黑龙江八一农垦大学 | Bacteria compsite inoculum used in lignin decomposition |
| CN105176868A (en) * | 2015-09-08 | 2015-12-23 | 福建省微生物研究所 | Complex bactericide for efficiently purifying sewage anaerobic outlet water of pig farm and preparation method and application thereof |
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| CN108587985A (en) * | 2018-06-14 | 2018-09-28 | 佛山市碧沃丰生物科技股份有限公司 | A method of degradation spirulina |
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