CN111826318A - Microcystis aeruginosa dissolving bacterium and application thereof - Google Patents
Microcystis aeruginosa dissolving bacterium and application thereof Download PDFInfo
- Publication number
- CN111826318A CN111826318A CN202010747863.0A CN202010747863A CN111826318A CN 111826318 A CN111826318 A CN 111826318A CN 202010747863 A CN202010747863 A CN 202010747863A CN 111826318 A CN111826318 A CN 111826318A
- Authority
- CN
- China
- Prior art keywords
- algae
- microcystis
- strain
- bacteria
- microcystis aeruginosa
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- 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
-
- 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
- C12R2001/07—Bacillus
-
- 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
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Hydrology & Water Resources (AREA)
- Environmental & Geological Engineering (AREA)
- Water Supply & Treatment (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to the technical field of microbial engineering, and particularly relates to a microcystis aeruginosa dissolving strain and application thereof, wherein the strain is heat-resistant Bacillus (Bacillus sporotherm modarans) with a preservation name of heat-resistant Bacillus Z-1; the invention also optimizes the culture system of the strain under the applicable condition, discusses the algae-lysing mechanism of the strain, researches the influence of parameters such as temperature, concentration of an added bacteria solution, pH value and the like on the algae-lysing efficiency, tests show that the strain has good dissolving and eliminating effects on the microcystis aeruginosa, mainly exerts the algae-lysing effect through extracellular products, has the highest algae-lysing efficiency when the light intensity is 1800-2200 lux at the temperature of 25-40 ℃, the light dark period is 10-15 h: 10-15 h and the pH value is 6-9, and can be applied to the treatment of microcystis aeruginosa polluted water.
Description
Technical Field
The invention belongs to the technical field of microbial engineering, and particularly relates to microcystis aeruginosa dissolving bacteria and a restoration application thereof to water bodies polluted by algae.
Background
Eutrophication of water causes mass propagation of algae, thereby causing outbreaks of water bloom and red tide, deteriorating water quality, affecting the function of water body and even threatening the safety of drinking water. The main manifestations of algae damage are: the algae cover the water surface in a large area to cause water quality corruption, so that fishes suffocate; many blue-green algae can produce algal toxins, which can cause damage to the nervous system of a human body, cause fright, dyspnea and even respiratory failure, and possibly cause diarrhea, vomiting and damage to organs such as liver and kidney; environmental indexes such as water transparency, dissolved oxygen and the like are influenced, so that water quality is deteriorated, and reasonable utilization of water resources is influenced.
The ecological problems caused by algae pollution are gradually attracting public attention. The currently common algae removal method mainly comprises three physical methods, a chemical method and a biological method, and the physical method cannot fundamentally solve the problems and has higher cost; the chemical algae removal method has quick response, but chemical agents are easy to generate secondary pollution and cause adverse effects on an ecosystem; the microbial algae control technology is a novel ecological restoration technology, has the advantages of high efficiency, environmental safety, no pollution and the like, and has wide application prospect.
In the 70 s of the 20 th century, people found that there was a good correlation between the content of chlorophyll of algae and the amount of algicidal bacteria in eutrophic water. The screening and application of the algicidal bacteria become a more active research direction in the field of biological algae control. At present, the separated bacteria with the algae-lysing effect mainly comprise vibrio, pseudomonas, alteromonas, bacillus, coccus and the like, and the main acting objects of the bacteria comprise freshwater or marine algae such as blue algae, dinoflagellate, diatom and the like.
The microcystis aeruginosa is one of the most common algae in eutrophic water bodies in China, an efficient microcystis aeruginosa dissolving bacterium is screened, and a culture application technical system of the microcystis aeruginosa dissolving bacterium is optimized, so that the microcystis aeruginosa dissolving bacterium is important for realizing green biological prevention and control of microcystis aeruginosa pollution, can assist in development of green treatment technology of water body eutrophication in China, and has wide application prospect.
Disclosure of Invention
The invention provides a microbial strain capable of being applied to water body remediation and treatment of algae pollution, aiming at solving the technical problem of serious algae pollution of water bodies.
Specifically, the invention relates to a heat-resistant bacillus Z-1 for dissolving microcystis aeruginosa, which is characterized in that: the strain is heat-resistant Bacillus (Bacillus sporotherm modurans) with the preservation name of Z-1 algicidal bacteria, the preservation unit is the common microorganism center of China Committee for culture Collection of microorganisms, the address is No. 3 Xilu No. 1 of Beijing Kogyang district, the preservation number is CGMCC NO:20070, and the preservation date is 11 days 06 months 11 in 2020.
The invention also relates to a microbial agent which is characterized by comprising the heat-resistant bacillus Z-1 microbial strain.
The invention also relates to application of the heat-resistant bacillus Z-1 or a microbial agent containing the heat-resistant bacillus Z-1 in treating microcystis aeruginosa.
Preferably, the heat-resistant bacillus Z-1 or the microbial agent containing the heat-resistant bacillus Z-1 is applied to the blue algae water bloom treatment; the blue algae is selected from one or more of microcystis aeruginosa, anabaena hyalopecuroides, cylindrospora sp.
Preferably, the application of the bacillus thermotolerans Z-1 or the microbial agent containing the bacillus thermotolerans Z-1 in the treatment of the microcystis aeruginosa is characterized in that: adding heat-resistant bacillus Z-1 into microcystis aeruginosa water according to the volume ratio of 5-10% of a microalgae liquid, and preferably adding heat-resistant bacillus Z-1 into the microcystis aeruginosa water at the temperature of 25-40 ℃ and the illumination intensity of 1800-2200 lux, preferably 1800lux, 1900lux, 2000lux, 2100lux and 2200 lux; the light-dark period is 10-15 h: 10-15 h, preferably 10-12 h: 10-12 h, and the pH value is 6-9.
Further preferably, before application, the heat-resistant bacillus Z-1 can be cultured for 18-24 h to logarithmic growth phase under the conditions that the temperature is 25-37 ℃ and the rotating speed of a shaking table is 100-280 r/min.
The invention also relates to a method for separating and screening the heat-resistant bacillus Z-1, which comprises the following steps:
screening of algicidal bacteria: standing activated sludge obtained from a second sewage treatment plant in Maanshan city, Anhui province, removing supernatant, inoculating 5mL of activated sludge into an LB culture medium, and culturing in a constant-temperature incubator at 30 ℃ and 150 rpm; then 5mL of culture solution is taken to be cultured in a new LB culture medium; adding the cultured bacterial liquid in logarithmic phase into the cultured microcystis aeruginosa, and culturing in an illumination culture box at 25 deg.C under illumination intensity of 2000lux for 12 h/12 h; shaking for 2 times every day, and observing the color change of the microcapsule algae; if the color of the algae is changed into yellow green or yellow white, and a small amount of brown precipitate exists at the bottom of the bottle, the existence of algae-lysing bacteria in the system is suggested;
and (3) separation of the algicidal bacteria: taking 1mL of supernatant in an algicidal bacteria system, sequentially diluting the supernatant with sterile distilled water according to concentration gradient, taking 100 mu L of diluted sample, uniformly coating a glass rod on an LB solid plate, inverting the plate and putting the plate into an incubator at 30 ℃ for culture, picking 10 single bacterial colonies with larger appearance morphology difference after the bacterial colonies grow out, inoculating the single bacterial colonies into a solid LB culture medium by the same method until a single purified bacterial colony is formed, inoculating the single bacterial colony into a slant culture medium and storing the slant culture medium in a refrigerator at 4 ℃;
primary screening of the algicidal bacteria: adding the bacterial solutions respectively cultured to logarithmic growth phases into the cultured microcystis aeruginosa, culturing under the conditions that the temperature is 30 ℃, the illumination intensity is 2000lux, the light-dark period is 12h:12h, observing the growth condition of the algae, primarily screening strains corresponding to the yellowed algae samples to obtain algicidal bacteria;
rescreening algicidal bacteria: inoculating the primary screening strain cultured to logarithmic phase into liquid cultured micro-cystic algae according to 10% of the volume ratio of the strain liquid, setting a culture medium with the same volume as a control group, and keeping the other treatment conditions the same; (ii) a Observing the color change of the water body, whether the microcystis is yellowed or not and whether the algae are aggregated or not every day; if the water body has yellowing phenomenon after the bacteria liquid is inoculated compared with a control group, the algae cells are reduced, and the bacteria is defined as having algae dissolving effect; the microcystis yellows in a period of time, compared with a control group, the microcystis cells have obviously reduced density and even disappear, but the microcystis cells begin to reproduce after a period of time is regarded as having inhibitory action but not complete;
determination of chlorophyll a of microcystis aeruginosa: chlorophyll a is selected as a growth index of microcystis aeruginosa in an algae-bacteria co-culture experiment, and the main operations are as follows: centrifuging a certain volume of algae solution (V1) at 5,000rpm at room temperature for 10min, collecting thallus, repeatedly freezing and thawing for 3 times to completely crack cells for extracting chlorophyll a; resuspend with a volume of 95% ethanol (V2) and place in a refrigerator at 4 ℃ overnight; centrifuging at 5,000rpm at room temperature for 10min, measuring absorbance of the supernatant at 665nm, 649nm and 750nm, calculating chlorophyll a content, and calculating algae-lysing rate according to the chlorophyll a content, wherein the strain with the highest algae-lysing rate is Bacillus thermostabilis Z-1.
More specifically, Bacillus thermotolerans Z-1 was isolated and selected as described in example 1.
The algicidal bacteria are explored through an algicidal mechanism, and both the algicidal bacteria filtrate and thalli can be used for dissolving algae, so that the algae dissolving mode of Z1 belongs to a way of releasing an algicidal substance to indirectly dissolve algae.
The algicidal bacteria can also have a good algicidal effect on microcystis aeruginosa cultured in a laboratory, and the algae lysing efficiency of the algicidal bacteria can change along with different environmental conditions.
The algae-lysing bacteria have very good killing effect on most of water-bloom blue-green algae including anabaena, cylindrosporium and microcystis, have good algae-lysing effect within the range of 18-35 ℃, can adapt to natural conditions of water bloom generation, and can be considered to be applied in actual water bloom treatment.
Drawings
The invention is further described below with reference to the accompanying drawings.
FIG. 1 shows the algal-lysing rate of different algal-lysing strains in example 1;
FIG. 2 is the algae-lysing rate of the Z-1 strain to Microcystis aeruginosa in example 3 under different temperature conditions;
FIG. 3 is the algae-lysing rate of Microcystis aeruginosa by the Z-1 strain of example 4 at various concentrations;
FIG. 4 shows the lysing rate of Microcystis aeruginosa by the Z-1 strain in example 5 at different pH values.
Detailed Description
The present invention will be further described with reference to the following examples. The described embodiments and their results are only intended to illustrate the invention and should not be taken as limiting the invention described in detail in the claims.
Example 1: separation, screening and identification of heat-resistant bacillus Z-1
1. Experimental Material
Activated sludge from the second sewage plant in Maanshan city, Anhui province
The Microcystis aeruginosa is purchased from Wuhan institute of Chinese academy of sciences, and cultured at 25 deg.C under the conditions of illumination intensity of 2000lx and light-dark ratio of 12h:12h after activation.
2. Culture medium
LB liquid medium: 10g/L of tryptone, 5g/L of yeast extract and 10g/L of NaCl.
LB solid medium: 10g/L of tryptone, 5g/L of yeast extract, 10g/L of NaCl and 20g/L of agar.
Culture of algae BG11 culture medium:
3. screening of algicidal bacteria activated sludge from a second sewage treatment plant in Maanshan city, Anhui province is kept still to remove supernatant, 5mL of activated sludge is taken and inoculated into an LB culture medium, and the activated sludge is cultured in a constant temperature incubator at 30 ℃ and 150 rpm. Then 5mL of the culture medium was added to a new LB medium for culture. Adding the cultured bacterial liquid in logarithmic phase into the cultured microcystis aeruginosa, and culturing in an illumination culture box at 25 deg.C under illumination intensity of 2000lux for 12 h/12 h. Shaking 2 times daily, and observing the color change of the microcapsule algae. If the algae color is changed into yellow green or yellow white, and a small amount of brown precipitate is formed at the bottom of the bottle, indicating that the algicidal bacteria possibly exist in the system.
4. Separating algicidal bacteria, taking 1mL of supernatant in an algicidal bacteria system, sequentially diluting the supernatant with sterile distilled water according to concentration gradient, taking 100 mu L of diluted sample, uniformly coating a glass rod on an LB solid flat plate, inverting the flat plate, putting the flat plate into an incubator at 30 ℃ for culture, after bacterial colonies grow out, selecting 10 single bacterial colonies with large appearance form difference, inoculating the single bacterial colonies into a solid LB culture medium by the same method until a single purified bacterial colony is formed, inoculating the single bacterial colony into a slant culture medium, and storing the slant culture medium in a refrigerator at 4 ℃.
5. And (3) primarily screening algicidal bacteria, namely adding the bacteria liquid which is respectively cultured to logarithmic growth phases into the cultured microcystis aeruginosa, culturing under the conditions that the temperature is 30 ℃, the illumination intensity is 2000lux and the light-dark period is 12h:12h, observing the growth condition of the algae, and primarily screening the strains Z-1, Z-3, Z-7 and Z-8 corresponding to the yellowed algae sample to obtain the algicidal bacteria.
6. Re-screening algicidal bacteria 4 primary-screened strains cultured to logarithmic phase are inoculated into liquid-cultured microcystis according to the liquid volume ratio of 10% bacteria and algae, and the strains without bacteria added are set as a control. Observing the color change of the water body, whether the microcystis is yellowed or not and whether the algae are aggregated or not every day. If the water body has yellowing phenomenon after the bacteria liquid is inoculated compared with a control group, the algae cells are reduced, and the bacteria is defined as having algae dissolving effect; the microcystis yellows in a period of time, the cell density of the microcystis is obviously reduced compared with that of a control group, even the microcystis disappears, but the microcystis starts to reproduce after a period of time is regarded as having the inhibition effect but is not complete.
7. Determination of chlorophyll a of microcystis aeruginosa, the chlorophyll a is selected as the growth index of the microcystis aeruginosa in an algae-bacteria co-culture experiment, and the main operations are as follows: centrifuging a certain volume of algae solution (V1) at 5,000rpm at room temperature for 10min, collecting thallus, repeatedly freezing and thawing for 3 times to completely crack cells for extracting chlorophyll a; resuspend with a volume of 95% ethanol (V2) and place in a refrigerator at 4 ℃ overnight; centrifuging at 5,000rpm at room temperature for 10min, measuring absorbance of the supernatant at 665nm, 649nm, and 750nm, and calculating chlorophyll a content according to the following formula:
Chl-a(mg/L)=[(A665-A750)×13.7-(A649-A750)×5.76]V2/V1
the algae-lysing rate is calculated according to the content of chlorophyll a, and the highest algae-lysing rate of the strain Z-1 is shown in figure 1.
8. Identification of lysophyta strains
The strain Z-1 is inoculated on an LB liquid culture medium and cultured for 48h under the conditions of 30 ℃ and 180 rpm. And (3) centrifuging the strain to collect thalli, extracting bacterial genome DNA by using a bacterial extraction kit, and sequencing by using a universal primer 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and a primer 1492R (5'-CGGGCGGTGTGTACAAG-3'). Compared with NCBI Blast search in GenBank, the similarity of the strain Z-1 of the invention and Bacillus sporotherm modems reaches 99 percent. According to the morphological characteristics, physiological and biochemical characteristics and 16S rDNA gene sequence characteristics of the strain Z-1, the strain Z-1 is identified to be heat-resistant Bacillus (Bacillus sporotherm modarans) and named as Bacillus sporotherm modans Z-1.
Example 2 discussion of the algae lysis method
Centrifuging the liquid culture Z-1 at 1200r/min, separating thallus from the supernatant, filtering the supernatant with a microporous filter membrane with the pore diameter of 0.22nm, washing the thallus twice with sterile distilled water, inoculating the filtrate into the liquid culture microcystis according to the volume ratio of 10%, adding the thallus into the cultured microcystis, and setting a blank algae solution as a control. Observing the color change of the water body, whether the microcystis is yellowed or not and whether the algae are aggregated or not every day. And simultaneously, measuring chlorophyll a of each group of microcystis aeruginosa.
Experiments show that the growth condition of algae is good in the experimental group of the single thalli and the algae liquid, while the algae turns yellow gradually and is agglutinated to a certain extent in the third day after the treatment of the thalli supernatant and the bacterial liquid experimental group, and brown precipitates can be seen at the bottom of the experimental group.
Wherein the content of chlorophyll a in blank control group is 2.6mg/L, the content of bacterial liquid is 0.6mg/L, the content of supernatant is 0.7mg/L, and the content of thallus is 2.3 mg/L. The chlorophyll a content is lowest after the supernatant of the thallus culture solution is treated, which indicates that the substances for inhibiting the growth of algae belong to extracellular products, and the algae dissolving mode belongs to a way of releasing the algae killing substances to dissolve algae indirectly.
EXAMPLE 3 Effect of temperature on the algae lysis Effect
The temperature is an important environmental condition for the growth metabolism and the energy metabolism of the strain, has great influence on the algae dissolving efficiency of the strain, and the determination of the optimal temperature range in the application process is the key for ensuring the strain to exert the maximum potential. The cultured thallus is inoculated with microcystis according to the proportion of 5 percent, and is respectively placed in a light incubator with the temperature of 20 ℃, 30 ℃, 35 ℃ and 40 ℃, and the algae dissolving rate of each experimental group is respectively measured to be 30 percent, 64 percent, 88 percent and 83 percent after one week as shown in figure 2.
The result shows that the algae dissolving efficiency is highest when the temperature is about 35 ℃, the outbreak of eutrophic algae generally occurs in summer with higher temperature in the practical application process, the water temperature is basically over 30 ℃, and the potential of algae dissolving bacteria can be better exerted.
Example 4 Effect of bacterial solutions of different concentrations on the algae lysis Effect
In the embodiment, 100mL of fresh microcystis aeruginosa is treated, the strain Z-1 is firstly cultured for 24h to logarithmic growth phase under the conditions that the temperature is 30 ℃ and the rotating speed of a shaking table is 150r/min, then the strain Z-1 is inoculated into 100mL of microcystis aeruginosa according to the liquid volume ratio of 2%, 5% and 10%, and LB culture medium is arranged as a control group. The system is placed under the conditions that the temperature is 35 ℃, the illumination intensity is 2000lux, the light dark period is 12h:12h for culture, and the algae-lysing rate is measured after one week. The algae-lysing rate of the algae solution treated by the above method was 60%, 85%, and 90% after 7d as shown in FIG. 3. The bacterial liquid adding amount volume ratio is 10%, and the algae dissolving effect is better.
Example 5 Effect of different pH on the algae lysing Effect
The pH is an important environmental condition for the growth and metabolism of the strain, and different pH can affect the algae dissolving effect of the strain. This example tested the algae lysing effect at different pH conditions, and cultured Microcystis aeruginosa at pH 5, 6, 8, 9, and 10, respectively, with the result that the growth of Microcystis was poor at pH 5 and 10. And (3) taking the algae solution with good growth condition under the conditions of pH 6, pH 8 and pH 9 for testing the algae dissolving bacteria. Taking 100mL of freshly cultured algae solution cultured under different pH conditions, taking the Z-1 strain cultured to logarithmic phase, inoculating the strain into 100mL of algae solution with different pH according to the volume ratio of 10% of the algae solution, and setting sterile water as a blank control. The system is placed under the conditions that the temperature is 35 ℃, the illumination intensity is 2000lux, the light dark period is 12h:12h for culture, and the algae-lysing rate is measured after one week. The algae solution treated by the above method has algae dissolution rates of 72%, 91% and 90% after 7d under the conditions of pH 6, pH 8 and pH 9, respectively, as shown in FIG. 4. The strain can be suitable for wide pH water bodies and can be used for repairing algae pollution.
Claims (7)
1. A heat-resistant bacillus Z-1 for dissolving microcystis aeruginosa is characterized in that: the strain is heat-resistant Bacillus (Bacillus sporotherm modurans) with the preservation name of Z-1 algicidal bacteria, the preservation unit is China general microbiological culture Collection center, the address is Beijing, China, the preservation number is CGMCC NO:20070, and the preservation date is 2020, 06 and 11 days.
2. A microbial agent characterized by comprising the Bacillus thermotolerant Z-1 strain according to claim 1.
3. An application of heat-resistant bacillus Z-1 or a microbial agent containing the heat-resistant bacillus Z-1 in treating microcystis aeruginosa.
4. Use according to claim 3, characterized in that: the heat-resistant bacillus Z-1 or the microbial agent containing the heat-resistant bacillus Z-1 is applied to the blue algae water bloom treatment; the blue algae is selected from one or more of microcystis aeruginosa, anabaena hyalopecuroides, cylindrospora sp.
5. Use according to claim 3, characterized in that: adding heat-resistant bacillus Z-1 into microcystis aeruginosa water according to the volume ratio of 5-10% of a microalgae solution, and treating the microcystis aeruginosa at the temperature of 25-40 ℃, the illumination intensity of 1800-2200 lux, the light-dark period of 10-15 h: 10-15 h and the pH value of 6-9.
6. Use according to claim 3, characterized in that: before application, the heat-resistant bacillus Z-1 can be cultured for 18-24 h to logarithmic growth phase under the conditions that the temperature is 25-37 ℃ and the rotating speed of a shaking table is 100-280 r/min.
7. The method for separating and screening B.thermotolerans Z-1 according to claim 1, which comprises:
screening of algicidal bacteria: standing activated sludge obtained from a second sewage treatment plant in Maanshan city, Anhui province, removing supernatant, inoculating 5mL of activated sludge into an LB culture medium, and culturing in a constant-temperature incubator at 30 ℃ and 150 rpm; then 5mL of culture solution is taken to be cultured in a new LB culture medium; adding the cultured bacterial liquid in logarithmic phase into the cultured microcystis aeruginosa, and culturing in an illumination incubator at 25 deg.C under illumination intensity of 2000lux for 12 h/12 h; shaking for 2 times every day, and observing the color change of the microcapsule algae; if the color of the algae is changed into yellow green or yellow white, and a small amount of brown precipitate exists at the bottom of the bottle, the existence of algae-lysing bacteria in the system is suggested;
and (3) separation of the algicidal bacteria: taking 1mL of supernatant in an algicidal bacteria system, sequentially diluting the supernatant with sterile distilled water according to concentration gradient, taking 100 mu L of diluted sample, uniformly coating a glass rod on an LB solid plate, inverting the plate and putting the plate into an incubator at 30 ℃ for culture, picking 10 single bacterial colonies with larger appearance morphology difference after the bacterial colonies grow out, inoculating the single bacterial colonies into a solid LB culture medium by the same method until a single purified bacterial colony is formed, inoculating the single bacterial colony into a slant culture medium and storing the slant culture medium in a refrigerator at 4 ℃;
primary screening of the algicidal bacteria: adding the bacterial solutions respectively cultured to logarithmic growth phases into the cultured microcystis aeruginosa, culturing under the conditions that the temperature is 30 ℃, the illumination intensity is 2000lux, the light-dark period is 12h:12h, observing the growth condition of the algae, primarily screening strains corresponding to the yellowed algae samples to obtain algicidal bacteria;
rescreening algicidal bacteria: inoculating the primary screening strain cultured to logarithmic phase into liquid cultured microcystis according to 10% of the volume ratio of the strain liquid, setting a culture medium with the same volume as a control group, and keeping the other treatment conditions the same; observing the color change of the water body, whether the microcystis is yellowed or not and whether the algae is agglutinated or not every day; if the water body has yellowing phenomenon after the bacteria liquid is inoculated compared with a control group, the algae cells are reduced, and the bacteria is defined as having algae dissolving effect; the yellowing phenomenon of the microcystis appears in a period of time, compared with a control group, the cell density of the microcystis is obviously reduced, even the microcystis disappear, but the microcystis starts to reproduce after a period of time and is regarded as having the inhibition effect but not complete;
determination of chlorophyll a of microcystis aeruginosa: chlorophyll a is selected as the growth index of microcystis aeruginosa in an algae-bacteria co-culture experiment, and the main operations are as follows: centrifuging a certain volume of algae solution (V1) at 5,000rpm at room temperature for 10min, collecting thallus, repeatedly freezing and thawing for 3 times to completely crack cells for extracting chlorophyll a; resuspend with a volume of 95% ethanol (V2) and place in a refrigerator at 4 ℃ overnight; centrifuging at 5,000rpm at room temperature for 10min, measuring absorbance of the supernatant at 665nm, 649nm and 750nm, calculating chlorophyll a content, and calculating algae-lysing rate according to the chlorophyll a content, wherein the strain with the highest algae-lysing rate is Bacillus thermostabilis Z-1.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202010747863.0A CN111826318B (en) | 2020-07-30 | 2020-07-30 | Microcystis aeruginosa dissolving bacterium and application thereof |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202010747863.0A CN111826318B (en) | 2020-07-30 | 2020-07-30 | Microcystis aeruginosa dissolving bacterium and application thereof |
Publications (2)
Publication Number | Publication Date |
---|---|
CN111826318A true CN111826318A (en) | 2020-10-27 |
CN111826318B CN111826318B (en) | 2022-03-29 |
Family
ID=72920352
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202010747863.0A Active CN111826318B (en) | 2020-07-30 | 2020-07-30 | Microcystis aeruginosa dissolving bacterium and application thereof |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN111826318B (en) |
Cited By (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108587981A (en) * | 2018-05-30 | 2018-09-28 | 西南大学 | Multi-functional molten algae streptomycete Streptomyces amritsarensis and its application |
CN113186131A (en) * | 2021-04-30 | 2021-07-30 | 广州绿曦生物科技有限公司 | Alga-lysing microbial agent and application thereof |
CN113583906A (en) * | 2021-07-21 | 2021-11-02 | 首都师范大学 | Application of pseudomonas B5 in algae removal |
CN113604407A (en) * | 2021-09-03 | 2021-11-05 | 中冶华天南京工程技术有限公司 | Composite microbial algaecide and preparation method and application thereof |
CN113773982A (en) * | 2021-08-10 | 2021-12-10 | 中国科学院生态环境研究中心 | Oligoxymonas strain KT48, algae-dissolving bacterium solution and preparation method and application thereof |
CN114634895A (en) * | 2022-04-07 | 2022-06-17 | 中山大学 | Bacillus cereus B1-XL001 with algae dissolving effect and application thereof |
CN115161231A (en) * | 2022-06-23 | 2022-10-11 | 轩凯生物科技(滁州)有限公司 | Bacillus coagulans with algae-lysing function and application thereof |
CN115386496A (en) * | 2022-09-23 | 2022-11-25 | 厦门大学 | Flagellates capable of ingesting microcystis aeruginosa and degrading microcystins and application |
CN115433691A (en) * | 2022-05-23 | 2022-12-06 | 西南大学 | Bacillus safensis T15 and application thereof |
CN116064276A (en) * | 2022-07-19 | 2023-05-05 | 南开大学 | Double-effect functional strain WL and application thereof in microcystis aeruginosa control and microcystin degradation |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102888353A (en) * | 2012-08-06 | 2013-01-23 | 常州大学 | Algicidal bacteria and method for removing microcystis aeruginosa |
CN103695342A (en) * | 2013-12-12 | 2014-04-02 | 上海交通大学 | Bacillus having alga-lysing activity and application thereof |
WO2018084895A1 (en) * | 2016-11-03 | 2018-05-11 | Marrone Bio Innovations, Inc. | Algicidal organisms |
-
2020
- 2020-07-30 CN CN202010747863.0A patent/CN111826318B/en active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102888353A (en) * | 2012-08-06 | 2013-01-23 | 常州大学 | Algicidal bacteria and method for removing microcystis aeruginosa |
CN103695342A (en) * | 2013-12-12 | 2014-04-02 | 上海交通大学 | Bacillus having alga-lysing activity and application thereof |
WO2018084895A1 (en) * | 2016-11-03 | 2018-05-11 | Marrone Bio Innovations, Inc. | Algicidal organisms |
CN110325042A (en) * | 2016-11-03 | 2019-10-11 | 马罗内生物创新公司 | Except algae organism |
Non-Patent Citations (2)
Title |
---|
P. SCHELDEMAN ET AL.: "Bacillus sporothermodurans and other highly heat-resistant spore formers in milk", 《OURNAL OF APPLIED MICROBIOLOGY 》 * |
胡长利等: "耐热芽孢杆菌(Bacillus sporothermodurans)的研究进展概述", 《食品安全质量检测学报》 * |
Cited By (18)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108587981A (en) * | 2018-05-30 | 2018-09-28 | 西南大学 | Multi-functional molten algae streptomycete Streptomyces amritsarensis and its application |
CN108587981B (en) * | 2018-05-30 | 2021-10-08 | 西南大学 | Streptomyces amyrosarensis and application thereof |
CN113186131A (en) * | 2021-04-30 | 2021-07-30 | 广州绿曦生物科技有限公司 | Alga-lysing microbial agent and application thereof |
CN113186131B (en) * | 2021-04-30 | 2023-10-27 | 广州绿曦生物科技有限公司 | Algicidal microbial agent and application thereof |
CN113583906A (en) * | 2021-07-21 | 2021-11-02 | 首都师范大学 | Application of pseudomonas B5 in algae removal |
CN113773982A (en) * | 2021-08-10 | 2021-12-10 | 中国科学院生态环境研究中心 | Oligoxymonas strain KT48, algae-dissolving bacterium solution and preparation method and application thereof |
CN113604407A (en) * | 2021-09-03 | 2021-11-05 | 中冶华天南京工程技术有限公司 | Composite microbial algaecide and preparation method and application thereof |
CN113604407B (en) * | 2021-09-03 | 2024-02-06 | 中冶华天南京工程技术有限公司 | Composite microbial algicidal fungicide and preparation method and application thereof |
CN114634895A (en) * | 2022-04-07 | 2022-06-17 | 中山大学 | Bacillus cereus B1-XL001 with algae dissolving effect and application thereof |
CN114634895B (en) * | 2022-04-07 | 2024-02-13 | 中山大学 | Bacillus cereus B1-XL001 with algicidal effect and application thereof |
CN115433691A (en) * | 2022-05-23 | 2022-12-06 | 西南大学 | Bacillus safensis T15 and application thereof |
CN115433691B (en) * | 2022-05-23 | 2024-03-15 | 西南大学 | Bacillus safoci Bacillus safensis T and application thereof |
CN115161231B (en) * | 2022-06-23 | 2023-03-31 | 轩凯生物科技(滁州)有限公司 | Bacillus coagulans with algae dissolving function and application thereof |
CN115161231A (en) * | 2022-06-23 | 2022-10-11 | 轩凯生物科技(滁州)有限公司 | Bacillus coagulans with algae-lysing function and application thereof |
CN116064276B (en) * | 2022-07-19 | 2023-07-21 | 南开大学 | Double-effect functional strain WL and application thereof in microcystis aeruginosa control and microcystin degradation |
CN116064276A (en) * | 2022-07-19 | 2023-05-05 | 南开大学 | Double-effect functional strain WL and application thereof in microcystis aeruginosa control and microcystin degradation |
CN115386496B (en) * | 2022-09-23 | 2023-10-27 | 厦门大学 | Dinoflagellates capable of feeding microcystis aeruginosa and degrading microcystin and application thereof |
CN115386496A (en) * | 2022-09-23 | 2022-11-25 | 厦门大学 | Flagellates capable of ingesting microcystis aeruginosa and degrading microcystins and application |
Also Published As
Publication number | Publication date |
---|---|
CN111826318B (en) | 2022-03-29 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN111826318B (en) | Microcystis aeruginosa dissolving bacterium and application thereof | |
CN103789232B (en) | The Pseudomonas stutzeri of one plant height effect process nitric wastewater and application thereof | |
CN105733998B (en) | Efficient denitrification strain with heterotrophic nitrification and aerobic denitrification capabilities | |
CN105132323B (en) | One plant of salt tolerant bacillus and its application in high-salt wastewater processing | |
CN107641609B (en) | Method for preparing flocculating agent by using compound microbial inoculum | |
CN101139140A (en) | Aerugo microcapsule algae degradation method using animalcule | |
CN105110480A (en) | Advanced treatment technology of high-salt-content degradation-resistant oil extraction sewage | |
CN113444661B (en) | Sphingobacterium neoformans and application thereof in wastewater dephosphorization | |
CN103421717A (en) | Bacillus cereus and application thereof | |
CN113604407A (en) | Composite microbial algaecide and preparation method and application thereof | |
CN116004485B (en) | Pseudomonas, microbial inoculum and method and device for treating dye wastewater by using same | |
CN103952359A (en) | Brevundimonas sp. and application thereof | |
CN111378592B (en) | Bacillus licheniformis and method for treating malodorous organic wastewater by using same to purify water | |
CN109609414B (en) | Unsymmetrical dimethylhydrazine degrading strain WP52 and application thereof | |
CN106591172B (en) | A kind of Rhodococcus ruber PTA-2 and its immobilization and application | |
CN109609405B (en) | Bacillus producing algae inhibiting active substance and use thereof | |
CN104694439B (en) | A kind of degrading crude oil bacterium and its application | |
CN116396902A (en) | Pseudomonas fish killing strain with algae dissolving capability and application thereof to red tide of red tide heterocurved algae | |
CN113913329B (en) | High-salt-tolerance COD-reduction bacterial strain, acquisition method and application | |
CN116515665A (en) | Pseudomonas capable of degrading microcystin, immobilized microbial agent and application | |
CN102206605B (en) | Exiguobacterium sp. with alga-lysing activity and application thereof in cyanobacterial bloom control | |
CN113249276B (en) | Bacillus cereus and application thereof | |
CN106754528B (en) | Pseudomonas flexi and application thereof | |
Al-Bader et al. | Subsurface associations of Acaryochloris-related picocyanobacteria with oil-utilizing bacteria in the Arabian Gulf water body: promising consortia in oil sediment bioremediation | |
CN110468066B (en) | Aerobic denitrifying strain and application thereof |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |