CN108018225A - Marine microorganism with excellent algicdal activity and the red tide removing method using the marine microorganism - Google Patents
Marine microorganism with excellent algicdal activity and the red tide removing method using the marine microorganism Download PDFInfo
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- CN108018225A CN108018225A CN201610953975.5A CN201610953975A CN108018225A CN 108018225 A CN108018225 A CN 108018225A CN 201610953975 A CN201610953975 A CN 201610953975A CN 108018225 A CN108018225 A CN 108018225A
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- C—CHEMISTRY; METALLURGY
- 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
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C—CHEMISTRY; METALLURGY
- 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
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/08—Seawater, e.g. for desalination
Abstract
This disclosure relates to a kind of marine microorganism with excellent algicdal activity and use its red tide removing method, and more specifically, it is related to a kind of Neptunomonas with excellent algicdal activity and belongs to marine microorganism bacterial strain, wherein the bacterial strain is fixed to the microorganism agent for being used to eliminate red tide of carrier, and includes the red tide removing method that at least one of the marine microorganism bacterial strain and the microorganism agent are introduced into the marine site that red tide occurs.
Description
Technical field
This disclosure relates to a kind of marine microorganism with excellent algicdal activity, and use the red of the marine microorganism
Damp removing method, more specifically, this disclosure relates in polycyclic cochlodinium sp (Cochlodinium polykrikoides, one kind
Cause the algae of red tide) in the case of have excellent selective algicdal activity marine microorganism bacterial strain, and use the sea
The red tide removing method of foreign microbial strains.
Background technology
Red tide typically refers to such phenomenon：(such as plant and animal planktonic organism, primary of the microorganism present in ocean
Animal and bacterium) while a large amount of propagation or accumulation, so as to change the color of seawater and marine organisms are adversely affected.Red tide
Fish and shellfish may be killed or be poisoned to phenomenon, cause a large amount of infringements to fishing ground, or even influence the mankind.Therefore, red tide phenomenon
Directly or indirectly cause serious problems.
All coastal waters are widely present red tide phenomenon in the world, and often regardless of season are occurred.Specifically,
Red tide plankton is changed into dinoflagellate class (dinoflagellates) from Diatomeae (diatoms), and tends to be densified.
Cause the biology of red tide can be, for example, protozoan, and phytoplankton such as diatom, dinoflagellate, blue-green alge, micro-
Algae etc..Specifically, only the biology of red tide is caused up to 50 kinds what South Korea found.According to the type of planktonic organism, the face of seawater
Color is not limited to become red, can also become burgundy, blueness, white and other colors.Analysis is from April, 2010 to 2013
After the red tide prewarning that July is sent by national academy of aquatic sciences, the red tide plankton frequently occurred wherein is identified.In Korea Spro
The biology for typically causing red tide that state occurs is polycyclic cochlodinium sp, and in the case of it, is sent out by national academy of aquatic sciences
118 red tide alarms are gone out.In addition, polycyclic cochlodinium sp is harmful dinoflagellate class, and known fishery can be caused significantly to damage.Remove
Outside this, Phaeocystis globosa (Chattonella marina), different cap algae (Heterocapsa triquetra), taper this gram
In general algae (Scrippsiella trochoides), Skeletonema Costatum (Skeletonemacostatum), Heterosigma akashiwo
(Heterosigma akashiwo), prorocentrum minimum (Prorocentrum minimum) etc. are confirmed as bearing a large amount of red tides
Have a responsibility for.
So far, the biological minimizing technology of red tide is caused to include chemical spray method, supercritical ultrasonics technology and ozone treatment, sea
Water detaches method, sedimentation, loess spraying etc..
Chemical spray method is most widely used that in these methods, that is, sprays copper sulphate (CuSO4), chlorine dioxide (ClO2)、
The method of Simanex etc..However, as relatively cheap so as to the copper sulphate of most widely used material, not particularly for harmful
Dinoflagellate class, has an effect on the marine organisms in addition to the biology for causing red tide, so that its problem is, in impacted waters
The toxicity and corrosivity of the other biological of middle existence.Further, since copper sulphate only has temporary transient effect, it may be necessary to makes repeatedly
Use copper sulphate.Since in the high alkalinity environment involved when algae red tide occurs, copper sulphate becomes unstable, it may be necessary to makes
With substantial amounts of copper sulphate, so as to reject the positive aspects of its its usage economy.
In Ebrean Registered Patent the 0407829th, a kind of red tide minimizing technology using ultrasonication is disclosed.Root
According to the red tide minimizing technology using ultrasonication, the biological cell of red tide is caused by ultrasonic destruction.It is in addition, smelly
Oxygen processing is that high-pressure ozone is introduced into the sea area that red tide occurs to neutralize the method for the toxicity as caused by red tide.However,
The stage that above two method is also not up to actually realized.
Seawater detach method and sedimentation using by whizzer, coagulating bath, mixing channel and pressure bank of cells into
Sedi-floator produces bubble to absorb the method with flotation red tide plankton, and red tide plankton is thus collected in seawater surface,
And seawater detaches method and sedimentation is widely used in South Korea.
In addition, in Ebrean Registered Patent the 0325396th, disclose a kind of red tide characterized by loess spraying and disappear
Except method.In other words, by loess (surface-active clay；Montmorillonite) seawater is sprayed to absorb and precipitate red tide plankton.In addition,
It make use of the inherent characteristic of loess for this, i.e. aluminium ion wherein included destroys the biological cell for causing red tide.Sprayed with loess
The red tide removing method that the method for spilling is characterized is widely used in South Korea.However, when spraying loess to seawater, floating material increase.
In fish farm and fishing zone (biology that wherein, there is colonization in lower floor), increased floating material may cause because blocking the fish gill exhales
Inhale exhaustion etc. and influence biology, it is therefore desirable to arouse attention.In addition, to cause red tide plankton to be stayed in after being killed marine for loess,
So as to be considered as the method that can cause secondary pollution in ocean.
A series of biological method has been had attempted to control red tide phenomenon, and for red tide phenomenon, the effect of bacterium
Highlight.
Therefore, it is contemplated that effectively apply situations below in the related art：Using for solving the above problems and effectively
Eliminate the combination of the new microbe of red tide.
The content of the invention
An aspect of this disclosure provides a kind of marine microorganism bacterial strain with excellent algicdal activity.
Another aspect of the disclosure provides a kind of microorganism agent for being used to eliminate red tide, it includes having excellent kill
The marine microorganism bacterial strain of algae activity.
Another aspect of the disclosure provide it is a kind of using with excellent algicdal activity marine microorganism bacterial strain or
Microorganism agent effectively eliminates the method for red tide.
According to one aspect of the disclosure, there is provided a kind of Neptunomonas with excellent algicdal activity belongs to ocean
Microbial strains.
Marine microorganism bacterial strain preferably includes Neptunomonas and belongs to (Neptunomonas sp.2R1) (preserving number
KACC81026BP)。
The marine microorganism bacterial strain preferably has the excellent choosing of the polycyclic cochlodinium sp in the algae to causing red tide
Selecting property algicdal activity.
According to another aspect of the disclosure, there is provided a kind of microorganism agent for being used to eliminate red tide, the wherein micro- life in ocean
Thing bacterial strain is fixed to carrier.
According to another aspect of the disclosure, there is provided a kind of red tide using the marine microorganism bacterial strain controls system
System.
According to another aspect of the disclosure, there is provided a kind of red tide removing method, this method are included to generation red tide
Sea area introduces the marine microorganism bacterial strain or microorganism agent.
Brief description of the drawings
According to reference to the described in detail below of attached drawing, above and other aspect, the feature of the disclosure will be more clearly understood
And advantage, in attached drawing：
Figure 1A is microphoto (multiplying power：More than 100 times, less than 1000 times), wherein, it is according to illustrative embodiments to kill
Phycomycete removes polycyclic cochlodinium sp (algae for causing red tide) with the time, and Figure 1B is microphoto (multiplying power：100 times), wherein, root
According to the phycomycete nutrient solution that kills of illustrative embodiments polycyclic cochlodinium sp (algae for causing red tide) is removed with the time；
Fig. 2 is to illustrate quantity marine microorganism, according to polycyclic cochlodinium sp cell with excellent algicdal activity
The figure of the algicdal activity of change；
Fig. 3 is illustrated in algicdal activity during the experiment, each figure grown with the time for killing phycomycete；
Fig. 4 be a diagram that killing algae bacterium Neptunomonas belongs to (Neptunomons sp.2R1) with especially red to causing
The figure of the selective algicdal activity of polycyclic cochlodinium sp in the algae of tide；
Fig. 5 is the figure for controlling the biology of red tide/ecological response system, which applies according to exemplary embodiment party
Formula kills segasso sea ocean microbial strains and ceramic monolith, it is illustrated that its result；
Fig. 6 is the lab diagram for controlling the biology of red tide/ecological response system, which applies according to exemplary reality
That applies mode kills segasso sea ocean microbial strains and ceramic monolith；And
What Fig. 7 illustrated in experimental group the polycyclic cochlodinium sp in the reactive tank and spout of (ceramics+bacterium) kills algae rate,
Wherein, the biology/ecological response according to illustrative embodiments for killing segasso sea ocean microbial strains and ceramic monolith is being applied
In system, phycomycete absorption is killed on carrier.
Embodiment
Hereinafter, embodiment of the present disclosure will be described with reference to the drawings.But the disclosure can lift in many different forms
Example explanation, and should not be construed as limited to particular implementation set forth herein.
According to an illustrative embodiments, there is provided there is the bacterial strain of excellent algicdal activity, wherein the bacterial strain quilt
Fixed to the microorganism agent on carrier, and use their red tide removing method.
According to an illustrative embodiments, the bacterial strain with excellent algicdal activity refers to that effectively eliminating (elimination) draws
Play the algae of red tide.Due to such effectiveness, bacterial strain according to illustrative embodiments can be widely used in controlling the work of red tide
Property such as suppress, eliminate.
According to illustrative embodiments, there is provided there is the marine microorganism bacterial strain of excellent algicdal activity,
Neptunomonas belongs to.Specifically, the bacterial strain preferably includes Neptunomonas and belongs to (Neptunomonas sp.2R1)
(preserving number KACC81026BP).The bacterial strain can have amphimicrobian characteristic.
It is micro- that each bacterial strain according to being included in the hybrid bacterial strain of an illustrative embodiments is preserved in South Korea's environment respectively
Biobank (KEMB).Specifically, Neptunomonas belongs to (Neptunomonas sp.2R1) and is preserved as KEMB 3-
825。
Belong to the marine bacteria of γ-mycetozoan as killing phycomycete to be widely known by the people.However, had according to illustrative embodiments
The Neptunomonas of algicdal activity belongs to α-mycetozoan, and the algicdal activity for belonging to such bacterium is seldom found relatively.
Marine microorganism bacterial strain according to illustrative embodiments with excellent algicdal activity is preferably
Neptunomonas belongs to (Neptunomonas sp.2R1).The bacterial strain is deposited in National Agricultural Science on 23rd in September in 2016 and grinds
Study carefully Korean agriculture Culture Collection (KACC), and obtain preserving number on October 13rd, 2016 and be
The preservation certificate of KACC81026BP.The address of KACC is South Korea, North Cholla 565-851, complete state prefecture, her west, 166 agricultures life
Order road.
Marine microorganism bacterial strain according to illustrative embodiments can have excellent relative to particularly microorganisms of red tide
In polycyclic cochlodinium sp algicdal activity.
According to an illustrative embodiments, there is provided for eliminating the microorganism agent of red tide, wherein, root as described above
Carrier is fixed to according to the marine microorganism bacterial strain with excellent algicdal activity of illustrative embodiments.It is exemplary according to one
Embodiment, when bacterial strain is fixed on carrier with applied to the marine site that red tide occurs, the bacterial strain can be stably distributed,
And the bacterial strain diluted degree in the seawater can be reduced.Therefore, its algicdal activity can continuously be maintained.
In this case, it is possible to bacterial strain is fixed by absorption.Specifically, the bacterial strain can be fixed by the following method
On carrier, wherein, the bacterial strain with ceramic monolith it is aerobic culture 4 it is small when.
Carrier according to illustrative embodiments is preferably included selected from admaic earth (red clay), stone sludge (stone
Sludge), spent bleaching clay (spent bleaching clay), electric arc furnaces (EAF) dust (electricarc furnace
Dust), in bloating shale (expanded shale), pelelith (volcanic stone) and flyash (coal fly ash)
At least one.
Specifically, carrier may include the admaic earth of 40wt% to 60wt%, the stone sludge of 20wt% to 40wt%, and
The spent bleaching clay of 5wt% to 20wt%, and preferably may also include electric arc furnaces (EAF) dust of 1wt% to 10wt%.It is highly preferred that
It may include the admaic earth of 55wt% to 65wt% according to the carrier of an illustrative embodiments, the stone of 20wt% to 30wt% is dirty
Mud, the spent bleaching clay of 5wt% to 15wt%, and the EAF dust of 2wt% to 8wt%, or may include 58wt% extremely
The admaic earth of 62wt%, the stone sludge of 23wt% to 27wt%, the spent bleaching clay of 8wt% to 12wt%, and 3wt% to 7wt%
EAF dust.Most preferably, the admaic earth of about 60wt% can be included according to the carrier of an illustrative embodiments, about
The stone sludge of 25wt%, the spent bleaching clay of about 10wt%, and the EAF dust of about 5wt%.
In carrier according to illustrative embodiments, its water absorption rate is preferably 15% to 25%.It is small in its water absorption rate
In the case of 15%, it may occur that the phenomenon that the bacterial strain residence time is reduced and processing time reduces.Its water absorption rate wherein
In the case of 25%, it may occur that the phenomenon that the hardness of carrier reduces.
In addition, in the case of carrier, its proportion is preferably 0.8 to 1.6.Its proportion is less than 0.8 situation wherein
Under, hardness is low, accordingly, it is possible to the problem of wherein carrier destruction occurs.In the case where its proportion is more than 1.6, hardness is good, but
It is possible that the problem of carrier is fixed in water.
In the case of carrier, its apparent porosity is preferably 17% to 26%.It is less than 17% in its apparent porosity
In the case of, it may occur that the problem of bacterial strain adsorbs.In the case where its apparent porosity is more than 26%, hardness may drop
It is low, and the problem of bacterial strain adsorbs may occur.
It is according to illustrative embodiments be used for eliminate red tide preferred microorganism agent can be used for eliminate due to comprising
The algae of polycyclic cochlodinium sp and the microorganism agent of red tide occurred, wherein, there is the polycyclic rotation ditch in the algae to causing red tide
The marine microorganism bacterial strain of the selective algicdal activity of algae, Neptunomonas belong to 2R1 (Neptunomonas sp.2R1, preservation
Number KACC81026BP) be fixed to the admaic earth including 60wt%, the stone sludge of 25wt%, 10wt% spent bleaching clay and
On the carrier of the EAF dust of 5wt%.
According to exemplary embodiment there is provided a kind of red tide removing method, including draw into the marine site that red tide occurs
Enter above-mentioned hybrid bacterial strain or microorganism agent according to illustrative embodiments.
Preferable red tide removing method according to illustrative embodiments can be eliminated due to comprising polycyclic cochlodinium sp
Algae and the method for red tide occurred, including introduce into the marine site that red tide occurs due to the algae comprising polycyclic cochlodinium sp micro-
Biological agent.
Hereinafter, the disclosure will be more fully described by embodiment.Following embodiments are provided to be only used for helping the disclosure
Understand, and the scope of the present disclosure not limited to this.
Embodiment
1. micro-biological samples gather
Select the coastal waters in the Tong Ying cities in great Han straits (South Korea south sea) four areas (Ma Shanwan, Zhenghai bay,
Pig island and floating fish pot), and seawater and deposit are collected from each area, as shown in table 1.In the case of seawater, due to molten
Solve that the situation of oxygen and temperature is different according to the depth of seawater, the distribution of microorganism is also considered as different.Therefore, collect simultaneously
Mixing is from the seawater with different depth position.
【Table 1】
Using water sample receiving flask each seawater samples of 20L are collected in each point.In cooler to be transported collected by storage
Seawater and collected deposit.
2. strain isolation
Seawater and marine sediment are collected in the marine site that red tide frequently occurs from South Korea's southern coast, with from its separating marine
Bacterium.
Based on collected sample, using various culture mediums to protect Algicidal microorganism.In the case of culture medium, for this
Using 10% ocean agar (Difco 2216, table 2), 10% R2A agar (MB cells, table 3), 100% ocean agar,
With 100% R2A agar.In addition, all culture mediums are formed according to the NaCl of 35 ‰ salinity 35g/L, divided with culture from seawater
From microorganism.
【Table 2】
The culture media composition of ocean bacterium solution 2216 (controlling pH is pH value 7.6 ± 0.2)
【Table 3】
The composition of R2A bacterium solutions (controlling pH is pH value 72 ± 02)
| Peptone | 0.5g |
| Yeast extract | 0.5g |
| The sour digest of casein | 0.5g |
| Glucose | 0.5g |
| Soluble starch | 0.5g |
| Dipotassium hydrogen phosphate | 0.3g |
| Magnesium sulfate | 0.024g |
| Sodium Pyruvate | 0.3g |
| D.W. | 1000ml |
It has been shown in table 4 in the case of aerobic condition, from the region for wherein collecting sample, and for each culture
Base separated number, and is shown in table 5 in the case of anaerobic condition, the region and its number.In aerobic condition and
Under anaerobic condition, sample is collected separately three times, and the sum of separated bacterial strain is respectively 3490 kinds and 1160 kinds.Except above-mentioned point
From bacterial strain, also use the bacterial strain selected under aerobic, amphimicrobian and anaerobic condition to test algicdal activity.
【Table 4】
【Table 5】
3. the selection and culture of red tide plankton
(1) selection of red tide plankton and culture medium
In order to select to occur most often at the microorganisms of red tide in coastal waters, according to existing file and the material survey frequency of red algae
Rate.In the case, the relevant paper of red tide delivered in South Korea, ocean and fishery science research institute, South Korea have been used for this
The database of marine science and technology research institute (KIOST) etc..Wherein, have selected frequency of occurrences quantity increases steadily, in the short time
Cause a wide range of serious infringement in cycle, and the limited polycyclic cochlodinium sp of control measure is used to kill algae experiment.The red algae of screening
From Seoul National University preservation.The algae of preservation is used in f/2 culture mediums (table 6).
【Table 6】
Table 7 illustrates the composition of the trace metal solutions of f/2 culture mediums, and the vitamin of the f/2 culture mediums shown in table 8 is molten
The composition of liquid.
【Table 7】
【Table 8】
Compared with cultivating the situation of red algae in the lab, when nature occurs, selected red algae is with the close of higher
Degree growth.Therefore, in the lab, culture medium and condition of culture are established to allow algae with High Density Cultivation.Algae is 20
DEG C, under conditions of 10000Lux, and cultivated under the concentration of 3.5% (w/v) NaCl.In addition, the condition of culture medium is divided into
Five kinds of conditions altogether, for example, 1. based on by autoclave sterilization, the seawater of filter paper using 0.2 μm filtering, pass through addition
Existing F/2 culture mediums prepared by F/2 compositions, 2. to the seawater addition F/2 combinations of the filtering without autoclave sterilization
The culture medium of the culture medium of thing, the 3. trace element (SL-6) of the vitamin solution to F/2 culture mediums addition 1ml and 1ml, 4. makes
The seawater filtered with 0.2 μm of filter paper, and the seawater of filter paper filtering is 5. not used, to carry out contrast experiment.
As a result of this experiment, polycyclic cochlodinium sp is found in F/2 culture mediums, and wherein to without high temperature and pressure
Grow more preferably in the culture medium of the seawater addition F/2 compositions of the filtering of sterilizing.In order to contamination-freely cultivate polycyclic cochlodinium sp,
Using by the F/2 culture mediums of autoclave sterilization for cultivating.
(2) screen
Algicdal activity is investigated using the mixed-culture medium containing separated marine microorganism bacterial strain and selected red algae.
In this case, use the method directly counted in microscope to alga cells.In order to screen the bacterial strain with algicdal activity,
In 24 orifice plates, cultivated in the Algae culture solution, ocean agar and R2A agar in 0.9mL and collect algae, then with 3.5%
NaCl is washed, and 3.5% sodium chloride is mixed with the inoculum of 0.1mL, its O.D. is adjusted to 1.0, to carry out mixing training
Support.24 it is small when after, to the mixture of special (Sedgewick-Rafter) room transfer 1ml of Sedgwick-pressgang, to use 1%
Lugol's solution fixes algae, and directly amount of algae is counted on microscope.
Algicdal activity is calculated with reference to equation 1.By screening, for algicdal activity high efficient strain, have selected with 50% with
On algicdal activity bacterial strain.On this basis, the experiment of algicdal activity has additionally been carried out confirming with 12 intervals when small.
Equation (1)
It has been shown in table 9 by process as described above in order to which excellent algicdal activity for polycyclic cochlodinium sp selects
The bacterial strain selected.Specifically, belong to the marine bacteria of γ-mycetozoan as killing phycomycete to be well known, but there is algicdal activity
Neptunomonas belong to α-mycetozoan, and the algicdal activity for belonging to the bacterium is seldom found relatively.
【Table 9】
4. the algicdal activity of algicdal activity high efficient strain is confirmed with the time
In order to be identified through 24 it is small when be mixed the algicdal activity for killing phycomycete of selection and change with time, pass through every 12
Hour counts biomass and checks the algicdal activity of each bacterial strain according to illustrative embodiments, and figure 1 illustrates.
In the case, the change of algicdal activity is confirmed by the change for the number for measuring polycyclic cochlodinium sp.
In addition, cultivated in Algae culture solution, ocean agar and R2A agar in 1.8mL and collect algae, Ran Houyong
3.5% NaCl washings, 3.5% sodium chloride is mixed with the inoculum of 0.2mL, its O.D. is adjusted to 1.0, to carry out
Mixed culture is so as to measure algicdal activity.Meanwhile in order to measure the quantity of the polycyclic cochlodinium sp changed, come pair using hemocytometer
The quantity of bacterium counts.In addition, the phenomenon that polycyclic cochlodinium sp is eliminated by killing algae bacterium is illustrated in Fig. 1.
Figure 1A is shown according to an illustrative embodiments, and (red tide is caused by killing the polycyclic cochlodinium sp of phycomycete elimination
Algae).Figure 1A be a diagram that confirms image that polycyclic cochlodinium sp is eliminated using microscope.Start for example, Figure 1A A are experiments
With kill after phycomycete processing 0 it is small when the image that catches, Figure 1A B are to kill the image caught when 12 is small after phycomycete processing, and Figure 1A C are to kill
The image caught when 24 is small after phycomycete processing.(caused red in addition, Figure 1B illustrates to kill phycomycete nutrient solution and eliminate polycyclic cochlodinium sp
The algae of tide).On the other hand, Figure 1B, which be a diagram that, kills the figure that phycomycete nutrient solution eliminates polycyclic cochlodinium sp (algae for causing red tide)
Picture.For example, Figure 1B A are to test the image for starting and killing and caught when 0 is small after phycomycete nutrient solution is handled, Figure 1B B are to kill phycomycete culture
The image caught when 12 is small after liquid processing, and Figure 1B C are to kill the image caught when 24 is small after phycomycete nutrient solution is handled.
In order to confirm the reappearance of experiment, further tested based on five bacterial strains with high algicdal activity, and
Thus, its result is described in figs. 2 and 3.
In the case, used bacterial strain is as described in table 9.In fig. 2, Neptunomonas belongs to
(Neptunomonas sp.2R1), killing phycomycete has a highest algicdal activity, and 98.9%.In figure 3, the growth rate of bacterial strain
For 8.29 × 106Cell/ml, and it is thus identified that the steady growth of bacterial strain.
5. the selection of the carrier according to illustrative embodiments suitable for bacterial strain
The place application process for killing phycomycete strain according to prior art can be that ground on the scene cultivates and to be sprayed kills phycomycete
Method, but described kill phycomycete strain and dilute in the seawater.Therefore, its effect often reduces.According to an illustrative embodiments,
As corresponding new method, algicdal activity high efficient strain, which is stablized, is attached to carrier, so as to avoid dilution effect and continuously keep killing
Algae activity.
As the example of the carrier for the bacterial strain according to illustrative embodiments with excellent algicdal activity, such as table
The cellular ceramic substrate of totally five types is prepared for described in 10.
Kill phycomycete to ceramic monolith to adhere to, to 4L optimal medium inoculating strain so as in 28 DEG C of optimal culture
At a temperature of cultivate three days.Each ceramic monolith is cleaned to remove powder, and Neptunomons (Neptunomonas sp.2R1)
The O.D. (600nm) of (Algicidal microorganism) is adjusted to 0.8.In addition, to strain cultured solution (about 108Cell/ml) addition 1.26kg
Ceramic aggregate, with allow cumulative volume become altogether about 3L, then mix at room temperature, with absorption 4 it is small when.
【Table 10】
In the ceramic monolith, using in extra large floating on water surface and the ceramic monolith 4 with optimal bacterial strain adsorption rate
Carrier carries out algicdal activity experiment.
In the case of the carrier of ceramic monolith 4, proportion is less than 1, and adsorption rate is more than 20%, and porosity is (apparent
Porosity) it is more than 60%.When absorption has the 2R1 bacterial strains of highest algicdal activity on the carrier in ceramic monolith 4, it was demonstrated that flat
2.31x10 is adsorbed7The bacterium of cell/g.Therefore, it is without interruption to kill phycomycete when applied to red tide or applied to place
Strain, and it is anticipated that maintain algicdal activity.
6. remove experiment using the red tide of bacterial strain according to illustrative embodiments
For using bacterial strain according to illustrative embodiments red tide remove experiment, be prepared for flow into groove, reactive tank and
Spout, and to flow into groove flow into 2961 cell/ml polycyclic cochlodinium sp.In reactive tank, arrange in ceramic monolith 2
The according to illustrative embodiments of absorption kills segasso sea ocean microbial strains 2R1.As a result, obtained in reactive tank
80.2% algae killing effect.Finally, the seawater by reactive tank that will be removed is contained in spout, treats finally to be arranged
Go out.As a result, it is last, obtain 97.2% algae killing effect.
Fig. 4 is to kill algae bacterium Neptunomonas categories (Neptunomons sp.2R1) to be confirmed to be with especially to causing
The figure of the selective algicdal activity of polycyclic cochlodinium sp in the algae of red tide.Specifically, Fig. 4 A show Neptunomons
(Neptunomons sp.2R1) is relative to the algicdal activity of polycyclic cochlodinium sp as a result, Fig. 4 B show Neptunomons
(Neptunomons sp.2R1) is relative to the algicdal activity of Phaeocystis globosa as a result, Fig. 4 C show Neptunomons
(Neptunomons sp.2R1) is relative to the algicdal activity of prorocentrum minimum as a result, Fig. 4 D show Neptunomons
(Neptunomons sp.2R1) is relative to the algicdal activity of different cap algae as a result, Fig. 4 E show Neptunomons
(Neptunomons sp.2R1) is relative to the algicdal activity of Heterosigma akashiwo as a result, Fig. 4 F show Neptunomons
(Neptunomons sp.2R1) relative to the algicdal activity of Skeletonema Costatum as a result, and Fig. 4 G show Neptunomons
(Neptunomons sp.2R1) relative to the algicdal activity of taper Si Kelipu algaes result.
Fig. 5 is the figure for controlling the biology of red tide/ecological response system, which applies according to exemplary embodiment party
Formula kills segasso sea ocean microbial strains and ceramic monolith, and which illustrates its result.
Fig. 6 is the figure for controlling the biology of red tide/ecological response system, which applies according to exemplary embodiment party
Formula kills segasso sea ocean microbial strains and ceramic monolith.
Fig. 7, which is illustrated, is applying the reality according to illustrative embodiments for killing segasso sea ocean microbial strains and ceramic monolith
In the case of testing group, polycyclic cochlodinium sp in reactive tank and spout kills algae rate.
As described above, according to illustrative embodiments, it can provide and not cause secondary pollution, without to other biological
Adverse effect, and the new microbe with excellent algicdal activity of microorganisms of red tide is effectively removed, and it is new micro- using this
The red tide removing method of biology.According to illustrative embodiments, microorganisms of red tide can be efficiently controlled to control in seawater or
The red tide occurred in fish farm etc..
Although being shown and having described with regard to illustrative embodiments above, to one skilled in the art show and
It is clear to, can be on the premise of the scope of the present invention defined in the appended claims not be departed from, various changes and modifications can be made.
Claims (7)
- A kind of 1. marine microorganism bacterial strain that Neptunomonas with excellent algicdal activity belongs to.
- 2. marine microorganism bacterial strain according to claim 1, wherein, the marine microorganism bacterial strain includes Neptunomonas belongs to 2R1 (Neptunomonas sp.2R1, preserving number KACC81026BP).
- 3. marine microorganism bacterial strain according to claim 1, wherein, the marine microorganism bacterial strain has to causing red tide Algae in polycyclic cochlodinium sp selective algicdal activity.
- 4. a kind of microorganism agent for being used to eliminate red tide, wherein, marine microorganism bacterial strain according to claim 1 is fixed To carrier.
- 5. according to claim 4 be used to eliminate the microorganism agent of red tide, wherein, the carrier include selected from admaic earth, At least one of stone sludge, spent bleaching clay, electric arc furnaces (EAF) dust, bloating shale, pelelith and flyash.
- A kind of 6. red tide control system using marine microorganism bacterial strain according to claim 1.
- 7. a kind of red tide removing method, including be introduced into the marine site that red tide occurs and appoint according in claim 1 to claim 5 Marine microorganism bacterial strain or microorganism agent described in one.
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