CN107475121A - The prevention and controls of harmful bacteria in a kind of microalga cultivation process - Google Patents
The prevention and controls of harmful bacteria in a kind of microalga cultivation process Download PDFInfo
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- CN107475121A CN107475121A CN201710852400.9A CN201710852400A CN107475121A CN 107475121 A CN107475121 A CN 107475121A CN 201710852400 A CN201710852400 A CN 201710852400A CN 107475121 A CN107475121 A CN 107475121A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/12—Unicellular algae; Culture media therefor
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
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Abstract
The present invention relates to a kind of prevention and controls of harmful bacteria in both culturing microalgae technical field, more particularly to microalga cultivation process.Specific aim improvement can be carried out to there is evil bacterium in microalga cultivation process in time.The prevention and controls of harmful bacteria in a kind of microalga cultivation process, including：Step 1) determines the harmful bacteria that the microalgae has polluted, and the physicochemical property determined by acquisition corresponding to harmful bacteria in microalgae carries out breeding process；Step 2) is cultivated to the microalgae, and in breeding process, the algae solution of the microalgae is monitored, and judges the physicochemical property for whether having corresponding to identified harmful bacteria in the algae solution；If it is determined that having the physicochemical property corresponding to identified harmful bacteria in the algae solution, then, physicochemical property of the step 3) determined by corresponding to harmful bacteria, specific aim improvement is carried out to the harmful bacteria.The embodiment of the present invention is used for both culturing microalgae.
Description
Technical field
The present invention relates to a kind of preventing and treating side of harmful bacteria in both culturing microalgae technical field, more particularly to microalga cultivation process
Method.
Background technology
At present, global economic development is restricted by energy scarcity.To realize sustainable economic development, microalgae biomass energy
Source receives significant attention as regenerative resource.But existing algae pilot scale culture is generally anti-in open pond or biology
Answer in device and carry out, in breeding process, especially summer, it is possible that a series of pollution, such as：It is germ contamination, primary
Asnimal pollution etc..Wherein germ contamination can occur at first in microalgae pollution course, and the bacterium of which part species can make microalgae
Yield decline, result even in large-scale culture several days, even round the clock between completely routed, the large-scale culture band to algae
Greatly endanger.
In order to prevent to produce germ contamination during algae pilot scale culture, at present, the incubation period generally before cultivation,
Sterilization treatment is carried out to nutrient solution and reactor, to avoid microalgae polluted bacteria in breeding process, also, existing scale
Culture is more to be cultivated using open or semi open model, can not avoid the pollution of bacterium completely in breeding process, and send out
Can only be when micro algae growth is affected also without the technology for differentiating harmful bacteria and harmless bacteria after raw pollution
Time is administered again, and dosage used in this when bactericide is larger, and microalgae that can be bad to growth conditions produces more not
Good influence.
It would therefore be highly desirable to seek a kind of method that harmful bacteria to during microdisk electrode carries out early stage improvement, to harmful
Bacterium carries out specific aim improvement, avoids harmful bacteria from administering caused microalgae yield not in time and declines, and can not be to harmful
Bacterium carries out the situation that microalgae yield caused by specific aim improvement declines and occurred.
The content of the invention
It is a primary object of the present invention to, there is provided the administering method of harmful bacteria in a kind of microalga cultivation process, by right
Harmful bacteria in microalga cultivation process is monitored in real time, can be entered in time to there is evil bacterium in microalga cultivation process
Row specific aim is administered, and is declined so as to avoid harmful bacteria from administering caused microalgae yield not in time, and can not be to having
Evil bacterium carries out the situation that microalgae yield caused by specific aim improvement declines and occurred.
To reach above-mentioned purpose, the present invention adopts the following technical scheme that：
The embodiment of the present invention provides a kind of prevention and controls of harmful bacteria in microalga cultivation process, including：
Step 1) determines the harmful bacteria that the microalgae has polluted, and obtain and determined in microalgae carries out breeding process
Harmful bacteria corresponding to physicochemical property；
Step 2) is cultivated to the microalgae, and in breeding process, the algae solution of the microalgae is monitored, and is judged
Whether there is the physicochemical property corresponding to identified harmful bacteria in the algae solution；
If it is determined that there is the physicochemical property corresponding to identified harmful bacteria in the algae solution, then,
Physicochemical property of the step 3) determined by corresponding to harmful bacteria, specific aim is carried out to the harmful bacteria and controlled
Reason.
Optionally, methods described also includes：
When the bacterial number increase in the algae solution for monitoring the microalgae, or, after the step 3), increase monitoring
Frequency.
Optionally, in microalgae carries out breeding process, the harmful bacteria that the microalgae has polluted is determined；Specifically include：
The microalgae is cultivated, and has bacterium in microalgae pollution, and/or, the biological increment of the microalgae is opened
When beginning to reduce, the algae solution of the microalgae is sampled, and the bacterium to occurring in algae solution carries out bacterium colony culture；
The bacterium from different bacterium colonies that culture is obtained is total to from sterile microalgae in different experimental groups respectively
With cultivation, if biomass decline occurs in the sterile microalgae in breeding process in the i-th experimental group, frustule color bleaches or turned yellow
And at least one of flocculation phenomenon occurs for frustule, it is determined that the bacterium in i-th experimental group is dirty for the microalgae
The harmful bacteria of dye, wherein, i is the natural number more than or equal to 1, and i-th experimental group is any one group in all experimental groups.
Optionally, the physicochemical property corresponding to harmful bacteria determined by acquisition；Specifically include：
Colonial morphology determined by observation corresponding to each harmful bacteria, to obtain the reason corresponding to each harmful bacteria
Change characteristic；Or
Gram's staining is carried out to identified each harmful bacteria, and observes each harmful bacteria after Gram's staining
Cellular morphology and the color that is presented, to obtain the physicochemical property corresponding to each harmful bacteria；Or
Determined dna sequence is carried out to each identified harmful bacteria respectively, and according to corresponding to each harmful bacteria
DNA sequence dna determine category belonging to each harmful bacteria, to obtain the physicochemical property corresponding to each harmful bacteria.
Optionally, the DNA sequence dna according to corresponding to each harmful bacteria, and being inquired about by ncbi database, it is each to determine
Category belonging to individual harmful bacteria.
Optionally, the bacterium from different bacterium colonies obtained will be cultivated respectively from sterile microalgae in different experimental groups
Before cultivate jointly, methods described also includes：
Determined dna sequence is carried out to the bacterium from different bacterium colonies respectively, and determines to come from according to determined dna sequence result
Whether the bacterium of different bacterium colonies is same bacterium, to filter out the difference cultivated jointly with sterile microalgae in all bacterium colonies
Strain, and according to the different strain filtered out, it is determined that being total to respectively to the bacterium from different bacterium colonies and the sterile microalgae
With the experimental group number of cultivation.
Optionally, the algae solution of the microalgae is monitored, and judges whether there is identified be harmful in the algae solution
Physicochemical property corresponding to bacterium；Specifically include：
The algae solution of the microalgae is sampled, and the bacterium polluted to the algae solution carries out bacterium colony culture, will cultivate
Colonial morphology corresponding to the colonial morphology and identified harmful bacteria of each bacterium colony obtained is contrasted；Or
The algae solution of the microalgae is sampled, and Gram's staining is carried out to the bacterium in the algae solution, by gram
Cellular morphology corresponding to each bacterium after dyeing and the color presented are contrasted with identified harmful bacteria；Or
Person,
The algae solution of the microalgae is sampled, and phylogenetic tree is established to the bacterium in the algae solution, according to being built
Vertical phylogenetic tree is to the 16SDNA sequences Design specific primers of identified harmful bacteria, according to designed specificity
Primer, identified harmful bacteria is tracked by PCR method.
Optionally, tracking is marked to identified harmful bacteria by fluorescence quantifying PCR method.
Optionally, the step 3) specifically includes：
According to the physicochemical property corresponding to identified harmful bacteria, the resistance to acids and bases of harmful bacteria determined by judgement,
And the difference between the resistance to acids and bases of the harmful bacteria and the cultivating microalgae, by adjusting the pH value of algae solution to being determined
Harmful bacteria administered；Or
Directly identified harmful bacteria is killed using the disinfectant of suitable dose.
The embodiment of the present invention provides a kind of prevention and controls of harmful bacteria in microalga cultivation process, by being supported in advance in foster algae
The harmful bacteria that the microalgae has polluted is determined during growing, and correspondingly corresponding to each harmful bacteria determined by acquisition
Physicochemical property, so, by during being cultivated to microalgae, being monitored in real time to the algae solution of the microalgae,
Judge in the algae solution physicochemical property for whether having corresponding to identified harmful bacteria, can be to occurring in the algae solution
Harmful bacteria in bacterium is monitored in real time, so, has identified harmful bacteria when monitoring in the algae solution
Physicochemical property when, according to the physicochemical property corresponding to identified harmful bacteria, specific aim is carried out to the harmful bacteria and controlled
Reason, harmful bacteria can be avoided to administer caused microalgae yield not in time and declined, and harmful bacteria can not be directed to
Property administer caused by microalgae yield decline situation occur, so as to harmful bacteria carry out early stage improvement, at utmost
The upper yield for improving microalgae.
Brief description of the drawings
In order to illustrate more clearly about the embodiment of the present invention or technical scheme of the prior art, embodiment will be described below
In the required accompanying drawing used be briefly described, it should be apparent that, drawings in the following description be only the present invention some
Embodiment, for those of ordinary skill in the art, on the premise of not paying creative work, can also be attached according to these
Figure obtains other accompanying drawings.
Fig. 1 is the flow signal of the prevention and controls of harmful bacteria in a kind of microalga cultivation process provided in an embodiment of the present invention
Figure；
Fig. 2 is a kind of harmful bacteria genome using 100 times of doubling dilutions provided in an embodiment of the present invention as the glimmering of template
The sensitivity curve figure of Fluorescent Quantitative PCR；
Fig. 3 is a kind of curve map of the specificity analysis of designed specific primer provided in an embodiment of the present invention；
Fig. 4 is that the fluorescence on the day of a kind of experimental group provided in an embodiment of the present invention and control group detect harmful bacteria respectively is determined
Measure PCR testing result figures.
Embodiment
Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out clear, complete
Site preparation describes, it is clear that described embodiment is only part of the embodiment of the present invention, rather than whole embodiments.It is based on
Embodiment in the present invention, those of ordinary skill in the art are obtained every other under the premise of creative work is not made
Embodiment, belong to the scope of protection of the invention.
The embodiment of the present invention provides a kind of prevention and controls of harmful bacteria in microalga cultivation process, referring to Fig. 1, including：
Step 1) determines the harmful bacteria that the microalgae has polluted, and obtain and determined in microalgae carries out breeding process
Harmful bacteria corresponding to physicochemical property；
Step 2) is cultivated to the microalgae, and in breeding process, the algae solution of the microalgae is monitored, and is judged
Whether there is the physicochemical property corresponding to identified harmful bacteria in the algae solution；
If it is determined that there is the physicochemical property corresponding to identified harmful bacteria in the algae solution, then,
Physicochemical property of the step 3) determined by corresponding to harmful bacteria, specific aim is carried out to the harmful bacteria and controlled
Reason.
Physicochemical property corresponding to harmful bacteria refers to characteristic of the harmful bacteria in terms of physics and chemistry.
The embodiment of the present invention provides a kind of prevention and controls of harmful bacteria in microalga cultivation process, by being supported in advance in microalgae
The harmful bacteria that the microalgae has polluted is determined during growing, and correspondingly corresponding to each harmful bacteria determined by acquisition
Physicochemical property, so, by during being cultivated to microalgae, being monitored in real time to the algae solution of the microalgae,
Judge in the algae solution physicochemical property for whether having corresponding to identified harmful bacteria, can be to occurring in the algae solution
Harmful bacteria in bacterium is monitored in real time, so, has identified harmful bacteria when monitoring in the algae solution
Physicochemical property when, according to the physicochemical property corresponding to identified harmful bacteria, specific aim is carried out to the harmful bacteria and controlled
Reason, harmful bacteria can be avoided to administer caused microalgae yield not in time and declined, and harmful bacteria can not be directed to
Property administer caused by microalgae yield decline situation occur, so as to harmful bacteria carry out early stage improvement, at utmost
The upper yield for improving microalgae.
In one embodiment of the invention, methods described also includes：
When the bacterial number increase in the algae solution for monitoring the microalgae, or, after the step 3), increase monitoring
Frequency.
The monitoring frequency to harmful bacteria can be increased, so as to enter to the harmful bacteria being likely to occur in breeding process
Row is administered in time.
In another embodiment of the present invention, in microalgae carries out breeding process, determine that the microalgae polluted is harmful thin
Bacterium；Specifically include：
The microalgae is cultivated, and has bacterium in microalgae pollution, and/or, the biological increment of the microalgae is opened
When beginning to reduce, the algae solution of the microalgae is sampled, and the bacterium to occurring in algae solution carries out bacterium colony culture；
The bacterium from different bacterium colonies that culture is obtained is total to from sterile microalgae in different experimental groups respectively
With cultivation, if biomass decline occurs in the sterile microalgae in breeding process in the i-th experimental group, frustule color bleaches or turned yellow
And at least one of flocculation phenomenon occurs for frustule, it is determined that the bacterium in i-th experimental group is dirty for the microalgae
The harmful bacteria of dye, wherein, i is the natural number more than or equal to 1, and i-th experimental group is any one group in all experimental groups.
Wherein, bacterium colony is to be bred by single bacterial cell in desired solid media surface or growth inside to certain journey
Degree, forms macroscopic daughter cell group.
In embodiments of the present invention, there is bacterium in microalgae pollution, refer to can clearly be seen that by micro- sem observation
There is germ contamination, and when the biological increment of the microalgae starts to reduce, illustrate to pollute bacterium in the microalgae, at this moment, lead to
Cross and the algae solution of microalgae is sampled, and bacterium colony culture is carried out to the bacterium in algae solution, can obtain all in the algae solution
The bacterium colony of bacterium, then the bacterium after bacterium colony culture is cultivated jointly with sterile microalgae respectively, it can determine the microalgae
Whether there is harmful bacteria in the bacterium of pollution, i.e. when the sterile microalgae in a certain experimental group does not grow, frustule substantially becomes
When at least one of obvious flocculation phenomenon phenomenon occur in leucismus Huang and microalgae, then it can determine that the bacterium in the experimental group is
Harmful bacteria, exemplary, when it is 3 to cultivate obtained bacterium colony, the bacterium in 3 bacterium colonies is respectively real first
Group, the second experimental group and the 3rd experimental group is tested to be cultivated jointly with sterile microalgae, wherein, the frustule in the second experimental group
There is frustule to bleach flavescence, then it is harmful bacteria to illustrate the bacterium in the second experimental group.Here, first, second, third be for
The name that experimental group is carried out is distinguished, the setting order and specific set-up mode not to experimental group cause to limit.
Under normal circumstances, the bacterium from different bacterium colonies is the bacterium of different strain, still, in the mistake that algae solution is coated
Cheng Zhong, the Different Individual of same bacterium may be coated on the diverse location on solid medium, therefore can form difference
Bacterium colony.
Therefore, in a preferred embodiment of the present invention, will cultivate the bacterium from different bacterium colonies that is obtained respectively with nothing
Before bacterium microalgae is cultivated jointly in different experimental groups, methods described also includes：
Determined dna sequence is carried out to the bacterium from different bacterium colonies respectively, and determines to come from according to determined dna sequence result
Whether the bacterium of different bacterium colonies is same bacterium, to filter out the difference cultivated jointly with sterile microalgae in all bacterium colonies
Strain, and according to the different strain filtered out, it is determined that being total to respectively to the bacterium from different bacterium colonies and the sterile microalgae
With the experimental group number of cultivation.
In embodiments of the present invention, by carrying out determined dna sequence to the bacterium from different bacterium colonies, and according to DNA sequences
Row measurement result, it can determine whether the bacterium from different bacterium colonies is same bacterium, so as to determine all bacterium colonies
Several different strains are included altogether, then are respectively cultivated different strains and sterile microalgae jointly, can be avoided same
Strain repeats to set experimental group, reduces the setting of experimental group.Exemplary, when it is 5 to cultivate obtained bacterium colony, for side
Just describe, this 5 different bacterium colonies are designated as the first bacterium colony, the second bacterium colony, the 3rd bacterium colony, the 4th bacterium colony and the 5th bacterium respectively in the future
Fall, determined dna sequence is carried out to the bacterium of this 5 different bacterium colonies respectively, when it is determined that the first bacterium colony and the 4th bacterium colony are same
Bacterium, and when the 3rd bacterium colony and the 5th bacterium colony are same bacterium, then it can determine have in the bacterium from this 5 different bacterium colonies
Three kinds of different strains, at this moment, it is possible to it is determined that being cultivated jointly to the bacterium from different bacterium colonies and the sterile microalgae
Experimental group number be three groups, i.e. the first experimental group can be in any one bacterium colony in the first bacterium colony and the 4th bacterium colony
Bacterium, the second experimental group can be the bacterium in the second bacterium colony, and the 3rd experimental group can be from the 3rd bacterium colony and the 5th
The bacterium in any one bacterium colony in bacterium colony, so as to avoid the repetition for same bacterium of the first bacterium colony and the 4th bacterium colony
The experimental group cultivated jointly with sterile microalgae is set, and the quantity of experimental group can be reduced to 3, reduce setting for experimental group
Put.Here, first, second, third is to distinguish the name of experimental group progress, and the setting not to experimental group is sequentially and specific
Set-up mode causes to limit.
Wherein, to before carrying out determined dna sequence from the bacterium of different bacterium colonies, methods described also includes：
DNA extractions are carried out to the bacterium from different bacterium colonies respectively, and performing PCR is entered using bacterial 16 S universal primer
(Polymerase Chain Reaction, PCR) expands.
PCR (Polymerase Chain Reaction, PCR) utilizes DNA 95 ° of high temperature Celsius in vitro
Time variation can become single-stranded, and primer is combined with single-stranded by the principle of base pair complementarity during low temperature (often 60 DEG C or so), then
Temperature regulating is to archaeal dna polymerase optimal reactive temperature (72 DEG C or so), and archaeal dna polymerase is along phosphoric acid to pentose (5'-3') side
To synthesis complementary strand.It is a kind of Protocols in Molecular Biology for being used to amplify the specific DNA fragmentation of amplification.
In another embodiment of the present invention, the physicochemical property corresponding to harmful bacteria determined by acquisition；Specifically include：
Colonial morphology determined by observation corresponding to each harmful bacteria, to obtain the reason corresponding to each harmful bacteria
Change characteristic.
Wherein, colonial morphology includes size, shape, edge, gloss, quality, color and transparency of bacterium colony etc..It is each
Kind bacterium forms fixed colony characteristicses under certain condition.
, can be to corresponding to each harmful bacteria by observing the colonial morphology corresponding to each harmful bacteria
Colonial morphology and the colonial morphology of other bacteriums make a distinction, so as to obtain the physics and chemistry spy corresponding to each harmful bacteria
Property.It is exemplary, when by carrying out bacterium colony culture to all bacteriums, and when to determine one of which bacterium be harmful bacteria, if
Bacterium colony corresponding to this bacterium is circular red petite, and bacterium colony moistens, be sticky, easily provoking, and remaining bacterium institute is right
The bacterium colony answered is, and bacterium colony is dried, and has fold, then it is circle that can obtain the physicochemical property corresponding to identified harmful bacteria
Red petite, and bacterium colony moistens, is sticky, easily provoking.
Or Gram's staining is carried out to identified each harmful bacteria, and that observes after Gram's staining each has
The cellular morphology of evil bacterium and the color presented, to obtain the physicochemical property corresponding to each harmful bacteria；
Gram's staining is widely used a kind of differential staining in bacteriology, the bacterium of no dyeing, due to its with
Surrounding environment index of refraction difference is very small, therefore extremely difficult observation under the microscope.Bacterium and environment form sharp contrast after dyeing, can be with
Form, arrangement and some architectural features of bacterium are clearly observed, and to taxonomic identification.Exemplary, when it is determined that all
When a kind of bacterium in bacterium is harmful bacteria, by carrying out Gram's staining to this bacterium, it can observe under the microscope
Shown face after to the shape (such as shaft-like, spherical, helical form) of this bacterium, and this bacterium progress Gram's staining
Color, compare, then can be obtained corresponding to identified harmful bacteria with positive formed of the shape and gram-negative of other bacteriums
Physicochemical property is Gram-positive or gram-negative rod bacterium, or spherical bacterium, or spirilla.
Or determined dna sequence is carried out to each identified harmful bacteria respectively, and according to each harmful bacteria institute
Corresponding DNA sequence dna determines the category belonging to each harmful bacteria, to obtain the physicochemical property corresponding to each harmful bacteria.
By carrying out determined dna sequence to identified harmful bacteria, and by determined dna sequence result and known DNA
Sequence is contrasted, and can determine the category belonging to identified harmful bacteria, so as to the harmful bacteria institute determined by
The general character that the accessory of category has, the physicochemical property determined by acquisition corresponding to harmful bacteria.Exemplary, when identified harmful
Category belonging to bacterium is bacillus, when the category belonging to other bacteriums belongs to for other, then can obtain identified harmful bacteria
With the physicochemical property corresponding to affiliated category, such as：When identified harmful bacteria is bacillus, it is Bacteriaceae
One category bacterium, it is strong to extraneous injurious factor resistance to obtain the harmful bacteria, and is gram-positive bacteria etc..
Wherein, in a preferred embodiment of the present invention, according to the DNA sequence dna corresponding to each harmful bacteria, and pass through
Ncbi database is inquired about to determine the category belonging to each harmful bacteria.
NCBI (National Center for Biotechnology Information) refers to US National biology skill
Art information centre.NCBI received the staff of molecular biology advanced training by the sequence submitted from each laboratory and
Same international nucleotides sequence database (EMBL and DDBJ) exchanges data and sets up database.With the arrangement of United States Patent and Trademark Office
So that the sequence information of patent is also integrated.
In one embodiment of the invention, the bacterium in the algae solution of the microalgae is monitored, and judges in the algae solution
Bacterium whether there is the physicochemical property of identified harmful bacteria；Specifically include：
The algae solution of the microalgae is sampled, and bacterium colony culture is carried out to the bacterium in algae solution, will cultivate what is formed
Colonial morphology corresponding to the colonial morphology of each bacterium colony and identified harmful bacteria is contrasted.
By carrying out bacterium colony culture to the bacterium in algae solution in incubation, and by the bacterium colony of each bacterium colony formed
Form is compared with identified harmful bacteria, and the harmful bacteria in the algae solution can be monitored in real time, exemplary
, when by compare, it is found that when a kind of bacterium colony in all bacterium colonies is identical with the colonial morphology of identified harmful bacteria, then may be used
To determine that the algae solution has the physicochemical property of identified harmful bacteria.
Or the algae solution of the microalgae is sampled, and Gram's staining is carried out to the bacterium in the algae solution, to leather
The cellular morphology of each bacterium after blue Albert'stain Albert and the color presented are contrasted with identified harmful bacteria.
By carrying out Gram's staining to the bacterium in algae solution in incubation, and to each bacterium after Gram's staining
Cellular morphology and the color that is presented be compared with identified harmful bacteria, equally can be to harmful in the algae solution
Bacterium is monitored in real time, exemplary, is found when by comparing：Have the bacterium after a kind of Gram's staining cellular morphology and
When the color presented is consistent with identified harmful bacteria, then it can determine that there is identified harmful bacteria in the algae solution
Physicochemical property.
Or the algae solution of the microalgae is sampled, and phylogenetic tree is established to the bacterium in the algae solution, according to
The phylogenetic tree established is to the 16SDNA sequences Design specific primers of identified harmful bacteria, according to designed spy
Specific primer, identified harmful bacteria is tracked by PCR method.
Sent out by establishing phylogenetic tree to the bacterium in the algae solution in incubation, and according to the system established
16SDNA sequences Design specific primer of the tree to identified harmful bacteria is educated, is passed through according to designed specific primer
PCR method is tracked to identified harmful bacteria, and equally the harmful bacteria in the algae solution can be monitored in real time.
Preferably, tracking is marked to identified harmful bacteria by fluorescence quantifying PCR method.Quantitative fluorescent PCR
It is (realtime fluores-cence quantitative PCR, RTFQ PCR), is by U.S. Applied
A kind of new quantitative test technology that Biosystems companies release, it is the specific spy by fluorescent dye or fluorescence labeling
Pin, tracking is marked to PCR primer, real time and on line monitoring course of reaction, product can be divided with reference to corresponding software
Analysis, calculate the initial concentration of testing sample template.Just start when, probe be incorporated in any one of DNA it is single-stranded on, PCR expand when,
5' ends -3' ends the 5 prime excision enzyme activity of Taq enzyme degrades probe digestion, separates reporter fluorescence group and quenching fluorescence group, from
And fluorescence monitoring system can receive fluorescence signal, i.e., a DNA is often expanded, just has a fluorescence molecule to be formed, realizes
The accumulation of fluorescence signal forms Complete Synchronization with PCR primer.The appearance of real-time fluorescence quantitative PCR, greatly simplifie quantitative inspection
The process of survey, and it is truly realized absolute quantitation.
In another embodiment of the present invention, the step 3) specifically includes：
According to the physicochemical property corresponding to identified harmful bacteria, the resistance to acids and bases of harmful bacteria determined by judgement,
And the difference between the harmful bacteria and the resistance to acids and bases of the microalgae, have by the pH value for adjusting algae solution to identified
Evil bacterium is administered.
Specific aim improvement can be carried out to identified harmful bacteria, and reduce the wound to the microalgae to the full extent
Evil.Exemplary, it is shaft-like when getting the physicochemical property corresponding to identified harmful bacteria by above-mentioned Gram's staining
Gramnegative bacterium (the soda acid tolerance range of the harmful bacteria is between 6.5-7.5), and corresponding microalgae is grid algae
When, then the pH value that can first adjust algae solution is 4.0 and kept for the regular hour, to be killed to the harmful bacteria, and
Processing is adjusted in the range of pH value to the normal growth of the grid algae of algae solution again after completing.
Or directly identified harmful bacteria is killed using the disinfectant of suitable dose.
Can equally specific aim improvement be carried out to identified harmful bacteria, and reduced to the full extent to the microalgae
Injury.Exemplary, when identified harmful bacteria being marked tracking by above-mentioned fluorescence quantifying PCR method, find algae
The concentration of harmful bacteria in liquid is the 10 of initial concentration3Left and right, then can be according to the concentration of the harmful bacteria, using suitable agent
The disinfectant of amount is killed to the harmful bacteria, the disinfectant of appropriate metrology be refer to just by harmful bacteria kill without
The dosage damaged to microalgae.
Wherein, the disinfectant can be sodium hypochlorite.
Hereinafter, will by embodiment, the present invention is described in detail, and by embodiment and control group to institute of the present invention
Caused technique effect is described in detail.
Embodiment 1
The embodiment 1 is with outdoor 0.2m2Runway it is pool cultivated intend Nannochloropsis oculata as experimental group, wherein, in following cultivation
During, the initial inoculation concentration for keeping intending Nannochloropsis oculata described in each experimental group is 3g/L, and cultivation number of days is 7 days,
Culture medium uses sea water medium, and light conditions are shone for outdoor natural light, and microdisk electrode situation measure is using the measure dry weight per 24h
Mode.
First, harmful bacteria is determined
In outdoor 0.2m2Nannochloropsis oculata is intended in inoculation in raceway pond, and initial concentration 3g/L, take under microscope has substantially had carefully
The algae solution of bacterium pollution, takes appropriate amount of sample, carries out the dilution of various concentrations using coubling dilution using sterilized water, and adjustment pollution is thin
The concentration of bacterium is 200/ml, and bacterial concentration is observed using blood counting chamber.
Take 200 microlitres to apply corresponding solid bacteria nutrition flat board, carry out bacterial clump culture in incubator at 33 DEG C.
The bacterium obtained after the above-mentioned carry out bacterium colony culture of picking, and the bacterium of each bacterium colony is activated, it will activate
Bacterium afterwards extracts DNA respectively, DNA sequencing is carried out after carrying out PCR method amplification using bacterial 16 S universal primer, according to being surveyed
DNA sequence dna all bacterium colony is subjected to division bacteria, and the bacterium of picking different strain, respectively with sterile plan Nannochloropsis oculata
It is 1 according to cell number：1 ratio, which is blended at 33 DEG C, carries out co-incubation, and observes the growing state that each group intends Nannochloropsis oculata,
If microalgae does not grow, either frustule, which substantially bleaches, turns yellow or produces microalgae flocculation phenomenon, it is determined that these bacteriums are harmful
Bacterium.
Conclusion：36 bacterium colonies are obtained altogether, 7 kinds of different strains are obtained after DNA sequencing, by by this 7 kinds of different bacterium
Kind carries out co-incubation with sterile plan Nannochloropsis oculata respectively, and acquisition has a kind of bacterium to be trained jointly with sterile plan Nannochloropsis oculata
Frustule bleaches behind foster 4 days.
2nd, the determination and checking of quantitative fluorescent PCR specific primer
All bacteriums are established into phylogenetic tree, the established phylogenetic tree of control is set to the 16SDNA of harmful bacteria
Count specific primer.
The sensitiveness of quantitative fluorescent PCR is determined using the target dna of 100 times of doubling dilutions as template respectively, wherein, fluorescence is fixed
It is 25 microlitres to measure PCR system, and quantitative fluorescent PCR enzyme is 12.5 microlitres, and primer is 1 microlitre, and template is respectively 2 microlitres, and fluorescence is fixed
Amount PCR reaction condition be：30s is kept at 95 DEG C first, afterwards, 95 DEG C of holding 5s, it is a circulation that 60 DEG C, which keep 15s, is followed
Ring 40 times；As a result it is shown in Figure 2, as shown in Figure 2：With the reduction of DNA dilution multiple proportions, the sensitiveness of quantitative fluorescent PCR is got over
By force, period is fewer, easier detection.Therefore, it can be deduced that：The detection signal of quantitative fluorescent PCR is relevant with DNA concentration,
Detection method is sensitive reliable, can carry out quantitative analysis.
Respectively using 2 nearer bacterium solution DNA of phylogenetic tree affiliation as template, it is determined that designed specific primer
Specificity.Wherein, quantitative fluorescent PCR system is 25 microlitres, and quantitative fluorescent PCR enzyme is 12.5 microlitres, and primer is 1 microlitre, mould
Plate is respectively 2 microlitres, and the reaction condition of quantitative fluorescent PCR is：First 95 DEG C keep 30s, afterwards, 95 DEG C holding 5s, 60 DEG C
It is a circulation to keep 15s, is circulated 40 times；As a result it is shown in Figure 3, as shown in Figure 3：Specific gene fluorescence intensity is non-specific
The 10 of property gene by fluorescence intensity6More than times, therefore, it follows that designed specific primer has very high specificity,
Detection available for early stage harmful bacteria.
3rd, in microalga cultivation process harmful bacteria monitoring and improvement
In outdoor 0.2m2Nannochloropsis oculata is intended in inoculation in raceway pond, initial concentration 3g/L, is supported to intending Nannochloropsis oculata
Grow, and according to the specific primer of above-mentioned determination in breeding process, by fluorescence quantifying PCR method to harmful thin in algae solution
Tracking is marked in bacterium, and harmful bacteria is monitored as experimental group；Meanwhile set under an equal cultivating condition
Control group, in breeding process by sediments microscope inspection by way of harmful bacteria is monitored.
Found by monitoring, during tracking is marked to harmful bacteria by the method for quantitative fluorescent PCR,
Harmful bacteria occurs in cultivation concentration after 3 days is the 10 of initial concentration3Times or so, show that institute occurs in experimental group after cultivating 3 days
The harmful bacteria of determination, at this moment, harmful bacteria is administered using 2ppm sodium hypochlorite, and by control group to harmful
When bacterium is monitored, just observe there is germ contamination after cultivating 5 days, at this moment, declining, which has occurred, in the yield of microalgae to become
Gesture, and need to administer harmful bacteria using 10ppm sodium hypochlorite.
Specifically, fluorescence intensity and pair of period that experimental group and control group are done on the day of harmful bacteria is detected respectively
It should be related to as shown in Figure 4, it is seen that, for control group compared with experimental group, the detection of harmful bacteria has certain hysteresis quality, it is impossible to and
When to harmful bacteria carry out specific aim improvement, the decline of microalgae yield can be caused.Wherein, by the micro- of experimental group and control group
Algae dry weight carries out periodic detection, can obtain result as shown in table 1 below.
Table 1
| Title | Experimental group yield (g/m2/ day) | Control group yield (g/m2/ day) |
| Cultivation 1 day | 7.61 | 7.7 |
| Cultivation 2 days | 8.1 | 8.03 |
| Cultivation 3 days | 7.92 | 7.79 |
| Cultivation 4 days | 7.88 | 6.98 |
| Cultivation 5 days | 8.06 | 4.88 |
| Cultivation 6 days | 7.87 | 0.35 |
| Cultivation 7 days | 8.13 | 0.75 |
| Average product | 7.94 | 5.21 |
As shown in Table 1：Experimental group can detect harmful bacteria in time, and after being administered using low dose of sodium hypochlorite,
The yield of microalgae is not had an impact so that bacterium is effectively controlled, and control group is when detecting harmful bacteria, bacterium
Quantity is very big, it is necessary to the sodium hypochlorite of larger dose is administered, and the yield of microalgae be subjected to it is a certain degree of
Influence, after being administered by the sodium hypochlorite of larger dose, a certain degree of injury can be produced to microalgae so that the production of microalgae
Amount drastically declines, and the average product of experimental group is more than control group.
Embodiment 2
Plate-type reactor of the embodiment 2 using outdoor light path as 10cm cultivates grid algae as experimental group, wherein, following
In breeding process, the initial inoculation concentration for keeping grid algae described in each experimental group is 3g/L, and cultivation number of days is 7 days, training
Support base and use BG11, light conditions are shone for outdoor natural light, and microdisk electrode situation measure is using the side that microalgae dry weight is determined per 24h
Formula.
First, harmful bacteria is determined
Specific method is same as Example 1, will not be repeated here.
Conclusion：41 bacterium colonies are obtained altogether, and the bacterium in 41 bacterium colonies and sterile grid algae are subjected to co-incubation respectively, can
To draw：There are two kinds of bacteriums to have cause evil effect to grid algae, one of which bacterium can make microalgae bleach after 5 days, another bacterium 4 days
After can make microalgae cell occur flocculation phenomenon.
2nd, the determination of the physicochemical property of harmful bacteria
Gram's staining is carried out to above-mentioned identified harmful bacteria, and it is thin corresponding to harmful bacteria determined by observation
Born of the same parents' form and the color presented.
Gram's staining result：Both the above bacterium is shaft-like gramnegative bacterium.The tolerance range of pH value exists
Between 6.5-7.5.
3rd, in microalga cultivation process harmful bacteria monitoring and improvement
Grid algae is inoculated with the plate-type reactor that outdoor light path is 10cm, initial concentration 3g/L, grid algae is cultivated,
And algae solution is sampled in breeding process, and Gram's staining is carried out to the bacterium in algae solution, as experimental group pair
Harmful bacteria is monitored；Meanwhile the control group under equal cultivating condition is set, by passing through microscope mirror in breeding process
The mode of inspection is monitored to harmful bacteria.
Found by monitoring, during being monitored by Gram's staining to harmful bacteria, cultivating the 3rd day just
Detection algae solution in there is identified harmful bacteria, at this moment, by adjust pH value to 4.0 keep 0.5h, afterwards adjust pH to
The method that microalgae normally cultivates scope, is administered to harmful bacteria.And when being monitored by control group to harmful bacteria,
Just observe there is germ contamination when cultivating the 5th day, at this moment, downward trend has occurred in the yield of microalgae, by adjusting pH
Value is administered to 4.0, and every the mode of 10min sampling microscopies to harmful bacteria, when bacterium of the microscopy without survival, altogether
Keep 1.5h, afterwards adjust pH normally cultivated to microalgae in the range of method harmful bacteria is administered.
It can be seen that control group, compared with experimental group, the detection of harmful bacteria has certain hysteresis quality, it is impossible in time to harmful
Bacterium carries out specific aim improvement, can cause the decline of microalgae yield.Wherein, by entering to the microalgae dry weight of experimental group and control group
Row periodic detection, result as shown in table 2 below can be obtained.
Table 2
| Title | Experimental group yield (g/m2/ day) | Control group yield (g/m2/ day) |
| Cultivation 1 day | 8.34 | 8.56 |
| Cultivation 2 days | 8.21 | 8.3 |
| Cultivation 3 days | 7.91 | 7.97 |
| Cultivation 4 days | 7.78 | 6.83 |
| Cultivation 5 days | 7.88 | 4.69 |
| Cultivation 6 days | 7.79 | 0.36 |
| Cultivation 7 days | 8.02 | 0.65 |
| Average product | 7.99 | 5.34 |
As shown in Table 2：Experimental group can detect harmful bacteria in time, and be controlled using short time reduction pH method
Reason so that bacterium is effectively controlled and do not had an impact to the yield of microalgae, and control group is when detecting harmful bacteria, bacterium
Quantity it is very big, it is necessary to which the method for reducing pH for a long time is administered, and the yield of microalgae has been subjected to a certain degree
Influence, after the method that pH is reduced by long-time is administered, a certain degree of injury can be produced to microalgae so that micro-
Drastically declining also occurs in the yield of algae, and the average product of the experimental group is more than control group.
Embodiment 3
The embodiment 3 is with outdoor 2m2The pool cultivated chlorella of runway as experimental group, wherein, in following breeding process,
The initial inoculation concentration for keeping chlorella described in each experimental group is 3g/L, and cultivation number of days is 7 days, and culture medium uses
BG11, light conditions are shone for outdoor natural light, and microdisk electrode situation measure is by the way of the measure microalgae dry weight per 24h.
First, harmful bacteria is determined
Specific method is same as Example 1, will not be repeated here.
Conclusion：35 bacterium colonies are obtained altogether, and carries out DNA sequencing respectively and obtains 5 kinds of different types of bacteriums, respectively by 5
The different types of strain of kind carries out co-incubation with sterile chlorella, it can be deduced that：There is a kind of bacterium to there is cause evil to make to chlorella
With frustule being made to bleach after 4 days.
2nd, the determination of the physicochemical property of harmful bacteria
The colonial morphology for observing above-mentioned identified harmful bacteria is to moisten, be sticky, is easily provoked, circular red petite.
3rd, in microalga cultivation process harmful bacteria monitoring and improvement
In outdoor 2m2Inoculation chlorella, initial concentration 3g/L, cultivates, and cultivating to chlorella in raceway pond
During algae solution is sampled, and bacterium colony culture is carried out to the bacterium in algae solution, by all bacterium colonies with it is identified harmful
The colonial morphology of bacterium is compared, and harmful bacteria is monitored as experimental group；Meanwhile equal cultivating condition is set
Under control group, in breeding process by sediments microscope inspection by way of harmful bacteria is monitored.
Found by monitoring, during being monitored by experimental group to harmful bacteria, just detected within the 3rd day in cultivation
Occur identified harmful bacteria in algae solution, at this moment, by using 2ppm sodium hypochlorite, harmful bacteria is administered.
And when being monitored by control group to harmful bacteria, just observe there is germ contamination when cultivating the 5th day, at this moment, microalgae
Yield there is downward trend, and need the sodium hypochlorite using 10ppm to administer harmful bacteria.
It can be seen that control group, compared with experimental group, the detection of harmful bacteria has certain hysteresis quality, it is impossible in time to harmful
Bacterium carries out specific aim improvement, can cause the decline of microalgae yield.Wherein, by entering to the microalgae dry weight of experimental group and control group
Row periodic detection, result as shown in table 3 below can be obtained.
Table 3
| Title | Experimental group yield (g/m2/ day) | Control group yield (g/m2/ day) |
| Cultivation 1 day | 8.22 | 8.21 |
| Cultivation 2 days | 8.04 | 8.13 |
| Cultivation 3 days | 7.91 | 7.86 |
| Cultivation 4 days | 6.78 | 6.73 |
| Cultivation 5 days | 6.54 | 4.58 |
| Cultivation 6 days | 7.07 | 0.53 |
| Cultivation 7 days | 7.02 | 0.47 |
| Average product | 7.37 | 5.22 |
As shown in Table 3：Experimental group can detect harmful bacteria in time, and after being administered using low dose of sodium hypochlorite,
The yield of microalgae is not had an impact so that bacterium is effectively controlled, and control group is when detecting harmful bacteria, bacterium
Quantity is very big, it is necessary to the sodium hypochlorite of larger dose is administered, and the yield of microalgae be subjected to it is a certain degree of
Influence, after being administered by the sodium hypochlorite of larger dose, a certain degree of injury can be produced to microalgae so that the production of microalgae
Amount drastically declines, and the average product of experimental group is more than control group.
In summary, the harmful bacteria polluted by determining the microalgae in microalga cultivation process in advance, and correspondingly
Physicochemical property determined by acquisition corresponding to each harmful bacteria, so, by the process cultivated to microalgae
In, the algae solution of the microalgae is monitored in real time, judges whether have corresponding to identified harmful bacteria in the algae solution
Physicochemical property, the harmful bacteria in the bacterium that occurs in the algae solution can be monitored in real time, so, work as monitoring
When there is the physicochemical property of identified harmful bacteria into the algae solution, according to the physics and chemistry corresponding to identified harmful bacteria
Characteristic, specific aim improvement is carried out to the harmful bacteria, harmful bacteria can be avoided to administer caused microalgae yield not in time
Decline, and the situation that microalgae yield caused by specific aim improvement declines can not be carried out to harmful bacteria and is occurred, so as to
Early stage improvement is carried out to harmful bacteria, improves the yield of microalgae to the full extent, meanwhile, and in the prior art in breeding process
Harmful bacteria is monitored by microscopy and compared, overcomes the defects of stronger by the hysteresis quality of microscopy in the prior art,
The early stage of pollution can just detect harmful bacteria, avoid harmful bacteria from having an impact the yield of microalgae, while can also avoid
Administering method caused by the increase of harmful bacteria quantity easily produces the defects of injuring to microalgae.
The foregoing is only a specific embodiment of the invention, but protection scope of the present invention is not limited thereto, any
Those familiar with the art the invention discloses technical scope in, change or replacement can be readily occurred in, should all be contained
Cover within protection scope of the present invention.Therefore, protection scope of the present invention should be based on the protection scope of the described claims.
Claims (9)
- A kind of 1. prevention and controls of harmful bacteria in microalga cultivation process, it is characterised in that including：Step 1) determines the harmful bacteria that the microalgae has polluted, and have determined by acquisition in microalgae carries out breeding process Physicochemical property corresponding to evil bacterium；Step 2) cultivates to the microalgae, and in breeding process, the algae solution of the microalgae is monitored, described in judgement Whether there is the physicochemical property corresponding to identified harmful bacteria in algae solution；If it is determined that there is the physicochemical property corresponding to identified harmful bacteria in the algae solution, then,Physicochemical property of the step 3) determined by corresponding to harmful bacteria, specific aim improvement is carried out to the harmful bacteria.
- 2. prevention and controls according to claim 1, it is characterised in thatMethods described also includes：When the bacterial number increase in the algae solution for monitoring the microalgae, or, after the step 3), increase the frequency of monitoring Rate.
- 3. prevention and controls according to claim 1, it is characterised in thatIn microalgae carries out breeding process, the harmful bacteria that the microalgae has polluted is determined；Specifically include：The microalgae is cultivated, and has bacterium in microalgae pollution, and/or, the biological increment of the microalgae starts to drop When low, the algae solution of the microalgae is sampled, and the bacterium to occurring in algae solution carries out bacterium colony culture；The bacterium from different bacterium colonies that culture is obtained is supported jointly from sterile microalgae in different experimental groups respectively Grow, if there is biomass decline in the sterile microalgae in breeding process in the i-th experimental group, frustule color bleach or turn yellow and At least one of flocculation phenomenon occurs for frustule, it is determined that the bacterium microalgae in i-th experimental group has polluted Harmful bacteria, wherein, i is the natural number more than or equal to 1, and i-th experimental group is any one group in all experimental groups.
- 4. prevention and controls according to claim 3, it is characterised in thatPhysicochemical property corresponding to harmful bacteria determined by acquisition；Specifically include：Colonial morphology determined by observation corresponding to each harmful bacteria, to obtain the physics and chemistry spy corresponding to each harmful bacteria Property；OrGram's staining is carried out to identified each harmful bacteria, and observes the thin of each harmful bacteria after Gram's staining Born of the same parents' form and the color presented, to obtain the physicochemical property corresponding to each harmful bacteria；OrDetermined dna sequence, and the DNA according to corresponding to each harmful bacteria are carried out to each identified harmful bacteria respectively Sequence determines the category belonging to each harmful bacteria, to obtain the physicochemical property corresponding to each harmful bacteria.
- 5. prevention and controls according to claim 4, it is characterised in thatInquired about according to the DNA sequence dna corresponding to each harmful bacteria, and by ncbi database, to determine each harmful bacteria institute The category of category.
- 6. prevention and controls according to claim 3, it is characterised in thatThe bacterium from different bacterium colonies that culture is obtained is supported jointly from sterile microalgae in different experimental groups respectively Before growing, methods described also includes：Determined dna sequence is carried out to the bacterium from different bacterium colonies respectively, and is determined according to determined dna sequence result from difference Whether the bacterium of bacterium colony is same bacterium, to filter out the different bacterium cultivated jointly from sterile microalgae in all bacterium colonies Kind, and according to the different strain filtered out, it is determined that being carried out respectively to the bacterium from different bacterium colonies with the sterile microalgae common The experimental group number of cultivation.
- 7. according to the prevention and controls described in claim any one of 1-6, it is characterised in thatThe algae solution of the microalgae is monitored, and judges whether have corresponding to identified harmful bacteria in the algae solution Physicochemical property；Specifically include：The algae solution of the microalgae is sampled, and the bacterium polluted to the algae solution carries out bacterium colony culture, and culture is obtained Colonial morphology corresponding to the colonial morphology and identified harmful bacteria of each bacterium colony obtained is contrasted；OrThe algae solution of the microalgae is sampled, and Gram's staining is carried out to the bacterium in the algae solution, by Gram's staining Rear cellular morphology corresponding to each bacterium and the color presented are contrasted with identified harmful bacteria；OrThe algae solution of the microalgae is sampled, and phylogenetic tree is established to the bacterium in the algae solution, according to what is established Phylogenetic tree draws to the 16SDNA sequences Design specific primers of identified harmful bacteria according to designed specificity Thing, identified harmful bacteria is tracked by PCR method.
- 8. prevention and controls according to claim 7, it is characterised in thatTracking is marked to identified harmful bacteria by fluorescence quantifying PCR method.
- 9. the prevention and controls according to right wants 1, it is characterised in thatThe step 3) specifically includes：According to the physicochemical property corresponding to identified harmful bacteria, the resistance to acids and bases of harmful bacteria determined by judgement, and Difference between the resistance to acids and bases of the harmful bacteria and the cultivating microalgae, have by the pH value for adjusting algae solution to identified Evil bacterium is administered；OrDirectly identified harmful bacteria is killed using the disinfectant of suitable dose.
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| CN101684447A (en) * | 2009-07-24 | 2010-03-31 | 新奥科技发展有限公司 | Method for effectively controlling enemy organisms in large-scale culture of microalgae |
| EP2292776A1 (en) * | 2000-07-31 | 2011-03-09 | Danisco US Inc. | Manipulation of genes of the mevalonate and isoprenoid pathways to create novel traits in transgenic organisms |
| CN104630067A (en) * | 2015-02-02 | 2015-05-20 | 新奥科技发展有限公司 | Pollution preventing and treating method for microalga breeding |
| CN106701589A (en) * | 2016-11-25 | 2017-05-24 | 新奥科技发展有限公司 | Method for treating pollution in microalgae culture process |
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|---|---|---|---|---|
| EP2292776A1 (en) * | 2000-07-31 | 2011-03-09 | Danisco US Inc. | Manipulation of genes of the mevalonate and isoprenoid pathways to create novel traits in transgenic organisms |
| CN101684447A (en) * | 2009-07-24 | 2010-03-31 | 新奥科技发展有限公司 | Method for effectively controlling enemy organisms in large-scale culture of microalgae |
| CN104630067A (en) * | 2015-02-02 | 2015-05-20 | 新奥科技发展有限公司 | Pollution preventing and treating method for microalga breeding |
| CN106701589A (en) * | 2016-11-25 | 2017-05-24 | 新奥科技发展有限公司 | Method for treating pollution in microalgae culture process |
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