CN109280627B - Method for rapidly and continuously culturing attached growth rhodopseudomonas photosynthetic bacteria - Google Patents
Method for rapidly and continuously culturing attached growth rhodopseudomonas photosynthetic bacteria Download PDFInfo
- Publication number
- CN109280627B CN109280627B CN201810694582.6A CN201810694582A CN109280627B CN 109280627 B CN109280627 B CN 109280627B CN 201810694582 A CN201810694582 A CN 201810694582A CN 109280627 B CN109280627 B CN 109280627B
- Authority
- CN
- China
- Prior art keywords
- photosynthetic bacteria
- rhodopseudomonas
- grass
- food waste
- water
- 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.)
- Active
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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a method for quickly and continuously culturing rhodopseudomonas photosynthetic bacteria growing in an attached state, which comprises the steps of firstly adding a water body containing phytoplankton into a light-transmitting container, adding grass or straw rich in cellulose into the light-transmitting container, then flatly paving solid food and beverage garbage mixed with monocalcium phosphate monohydrate and ferric citrate on the grass or straw, then adding a proper amount of magnesium hydrogen phosphate trihydrate after green attached algae appear, culturing for 19-25 days to ensure that a large amount of rhodopseudomonas photosynthetic bacteria appearing dark red or blood red are attached to the inner wall of the container, scraping the rhodopseudomonas photosynthetic bacteria attached to the inner wall, and then continuously adding the food and beverage garbage mixed with salts for culturing. The method can rapidly convert organic matters into photosynthetic bacteria of Rhodopseudomonas, promote harmless treatment and utilization of water body nutrient substances, accelerate conversion of water body nutrient substances, and directly use the formed photosynthetic bacteria in breeding industry, sewage purification or new energy technology.
Description
Technical Field
The invention relates to a method for culturing water body microorganisms, in particular to a method for quickly and continuously culturing attached growth rhodopseudomonas photosynthetic bacteria.
Background
Photosynthetic Bacteria (Photosyntetic Bacteria) are a generic term for a class of prokaryotes with primitive light energy synthesis systems, which are capable of anaerobic, anaerobic Photosynthetic processes, and are a class of gram-negative Bacteria. Currently, the most common photosynthetic bacteria species used in the fields of sewage treatment, aquaculture, and the like are Rhodopseudomonas (Rhodopseudomonas) of rhodospirillaceae, which belongs to the kingdom of prokaryotes, phototrophic prokaryotes, rhodobacter, in the class of microbial classification. The rhodopseudomonas photosynthetic bacteria have the main characteristics that: can resist lower temperature, and can not die even if frozen; can resist higher salinity; it is rich in crude protein, vitamin B group, carotenoid and coenzyme Q.
Rhodopseudomonas photosynthetic bacteria play a role in various aspects such as aquaculture, sewage treatment, new energy research and the like, and mainly comprise: (1) purifying water quality; (2) the oxygen consumption of organic matters is reduced; (3) a feed for zooplankton; (4) can be used as feed additive; (5) hydrogen gas generation, etc.
At present, the common method for culturing the rhodopseudomonas photosynthetic bacteria is to use an artificially configured culture medium, and in addition, related researches have found a plurality of large-scale culture methods for the rhodopseudomonas photosynthetic bacteria, wherein the culture raw materials can be compound feed, food waste and culture media configured by a plurality of chemical substances. These methods mainly utilize the characteristic that photosynthetic bacteria of the genus Rhodopseudomonas can degrade and utilize organic substances, but when the photosynthetic bacteria are cultured by directly using compound feed or food waste, the organic substances are required to rot under the condition of a proper amount of oxygen, so that the required culture time is long (at least more than 35 days), while when a culture medium is prepared by using a plurality of chemical substances, the culture time is short, but a plurality of chemical raw materials are required, and the preparation ratio of each substance is strictly required. The research of immobilizing the photosynthetic bacteria of the rhodopseudomonas genus is also available, which mainly utilizes some solid materials to make into micro-particles, and the rhodopseudomonas adheres to and grows on the micro-particles, and the micro-particles adhered with the rhodopseudomonas can be suspended in a water body under the condition of certain hydrodynamic force, so as to achieve the purpose of promoting the growth of the photosynthetic bacteria; the media selected for use in these studies is typically media prepared from a wide variety of chemical materials commonly used.
Disclosure of Invention
The invention aims to provide a method for quickly and continuously culturing the rhodopseudomonas photosynthetic bacteria growing in an attached state aiming at the technical defects that the existing large-scale culture method of the rhodopseudomonas photosynthetic bacteria needs long time or needs a plurality of chemical raw materials, and is more beneficial to the stable, quick and large-scale culture of the rhodopseudomonas photosynthetic bacteria by combining the existing large-scale culture method of the rhodopseudomonas photosynthetic bacteria.
In order to achieve the technical purpose, the invention designs a method for quickly and continuously culturing attached and growing rhodopseudomonas photosynthetic bacteria, which is characterized by comprising the following steps:
s1, placing the light-transmitting container in a glass greenhouse or an outdoor natural illumination environment, wherein the environmental temperature change range of the day and the night is 15-45 ℃, adding phytoplankton water into the light-transmitting container, and adding grass or straws rich in cellulose into the light-transmitting container to ensure that the grass or straws float on the upper layer of the water;
s2, adding the solid food waste mixed with the salt mixture to grass or straws, wherein the amount of the grass or straws added with the rich cellulose in the step S1 is enough to prevent the solid food waste from sinking into water and just submerge the solid food waste in the water;
s3, culturing for 3-5 days, and enabling the water body to become dark green and even brown green;
culturing for 6-10 days, and growing green attached algae in the area where the upper part of the inner wall of the container is close to the grass;
culturing for 11-14 days, enlarging the area of the deep green attached algae on the upper part of the inner wall of the container, and adding magnesium hydrogen phosphate trihydrate (MgHPO) when the deep green attached algae grows4·3H2O), the addition amount is 4-15 g;
s4, culturing for 15-18 days, wherein dark red attachments appear in green attached algae on the upper part of the inner wall of the container, and the dark green attached algae gradually turns into dark brown;
culturing for 19-25 days, wherein dark brown attachments on the inner wall of the container are completely changed into dark red or blood red rhodopseudomonas photosynthetic bacteria, and the area of the upper part of the inner wall of the container, which is close to the grass, can be changed into dark green or continuously keep dark red;
s5, scraping the rhodopseudomonas photosynthetic bacteria attached and grown in the step S4 by cloth under the condition that water in the container is not discharged; and then continuously adding the food wastes mixed with salts on the grass or the straws in the container, wherein the adding amount is not more than that of the previous adding amount, and magnesium hydrogen phosphate trihydrate is not needed to be added in the subsequent process, so that continuous multiple culture is realized until the grass or the straws therein are rotted and can not support the degradation of the food wastes on the water surface.
Further, step S1 is performed in a warm season from late 4 to late 11 months.
Further, in the step S2, the solid food waste and the water are added in a ratio of 15-65 g:1L, the thickness of the food waste stacked on the grass or straw is not more than 3cm, and the solid food waste is completely submerged in water.
Further, in the step S2, the salt mixture includes monocalcium phosphate monohydrate [ Ca (H)2PO4)2·H2O]And ferric citrate, wherein the mass addition proportion of the monocalcium phosphate monohydrate is 0.25-0.75% of the mass of the solid food waste, and the mass addition proportion of the ferric citrate is 0.1-0.25% of the mass of the solid food waste.
Further, in step S2, the solid food waste is food waste remaining after removing non-food substances, and includes some or all of rice, wheaten food, fish, meat, eggs, vegetables, fruits, salt, seasoning for making dishes, bones, eggshells, and the like, and mainly starch substances such as rice, wheaten food, and the like, so as to ensure that liquid water in the food waste is as little as possible, and the food waste is spread out and air-dried for 1 hour for later use.
Further, in the step S1, the material of the transparent container is common colorless transparent glass or transparent organic glass or white transparent plastic or colorless transparent plastic.
Further, in the step S1, the grass or straw is rice straw or grass or wheat straw for green lawn, which can keep the solid food waste on the water surface without sinking into the water bottom after being piled up to a certain thickness.
Further, in the step S4, the maximum thickness of the attached and grown photosynthetic bacteria of the genus Rhodopseudomonas can reach about 1-2 mm, and tap water is added at any time during the cultivation period to supplement the water consumed by evaporation.
Further, in the step S1, the total nitrogen concentration in the fresh water body containing phytoplankton which is initially added is 2-30 mg/L, and the chlorophyll a concentration is 30-2700 mug/L.
In the technical scheme of the invention, 5 conditions of addition of salts in the solid food waste, addition proportion of the transparent container, the solid food waste and water body rich in phytoplankton, temperature range, degradation of the solid food waste on the water surface and the like are the key for realizing the purpose of the invention. The purpose of adding hay in the container is to enable the food waste to be located on the water surface with floating grass as a support, so that the food waste can be easily contacted with oxygen in the air and can be quickly decomposed and degraded.
Calcium dihydrogen phosphate monohydrate and ferric citrate are added into the solid food waste, so that the solid food waste has the main effects of promoting the degradation of the food waste and supplementing metal cations such as calcium, iron and the like, and the requirement of the subsequent attachment and growth of a large amount of photosynthetic bacteria of the rhodopseudomonas is met.
The solid food waste can generate the condition of simultaneous odor and vinasse odor in the degradation process. When the adding proportion of the solid food waste and water is lower than 15g:1L, the maintenance time of obvious vinasse taste generated by the food waste after being degraded on the water surface is longer, the concentration of nutrient salts such as nitrogen, phosphorus and the like in the water body can support the water body to generate green attached algae after the food waste is rotted, but the green attached algae is not easily converted into red attached bacteria; when the adding ratio of the solid food waste to water is more than 65g:1L, or the stacking thickness of the food waste on grass is not more than 3cm, the food waste can generate heavy odor (mainly due to ammonia gas generated by degradation) after being degraded on the water surface, the whole water body is easy to lack of oxygen, the decay and degradation time of the solid food waste can be longer, and after the time for the subsequent water body to grow green attached algae can be pushed, the time for the whole red bacteria to attach can be pushed back to influence the time efficiency for cultivating the rhodopseudomonas photosynthetic bacteria.
Adding magnesium hydrogen phosphate trihydrate (MgHPO) which is difficult to dissolve in water when the deep green attached algae grows4·3H2O) because it can be used under anaerobic or hypoxic, meta-acidic conditionsA small amount of magnesium ions and phosphate ions are released by slow reaction, so that sufficient magnesium element can be provided for rhodopseudomonas rich in bacteriochlorophyll, and the content of other ions in a water body is not influenced. Magnesium hydrogen phosphate trihydrate can be replaced by soluble magnesium sulfate heptahydrate, only the magnesium sulfate heptahydrate is soluble, the salinity of a water body can be changed when the magnesium hydrogen phosphate trihydrate is added in a large amount, the magnesium hydrogen phosphate trihydrate can be precipitated at the bottom of the water body, salt powder is fished from the bottom of a glass jar after red bacteria grow to the inner wall, and the red bacteria can be obviously observed to be attached to and grow in the salt.
When the environmental temperature change range of the glass greenhouse or the outdoor natural illumination environment in the daytime and at night is 15-45 ℃, the temperature is higher and the illumination is abundant in warm seasons from early summer to late autumn, and the appropriate temperature and illumination conditions can be ensured. In the case of a winter season or a spring season where the temperature is low, the degradation of the solid food waste is slowed, and although green algae grow, it takes a longer time for the attachment of red bacteria to occur.
In the technical scheme of the invention, in the step S2, the total nitrogen concentration in the added fresh water body rich in phytoplankton is 2-30 mg/L, the chlorophyll a concentration is 30-2700 mug/L, and common natural fresh water bodies easily meet the range; and the high-concentration phytoplankton water generated after the organic matter is decayed can be selected, wherein the total nitrogen concentration can be as high as 30mg/L, and the phytoplankton concentration can be as high as 2700 mug/L. The purpose of selecting a certain amount of phytoplankton in the water body is to increase the oxygen content in the water body through the photosynthesis of the phytoplankton in the water body and provide an initial microbial community with rich diversity, thereby being beneficial to the rotting and degradation of the food waste at the initial stage of culture.
Secondly, the invention also relates to the application of the photosynthetic bacteria generated by the method in the breeding industry. The photosynthetic bacteria obtained by the invention can be used for treating aquaculture residual feed or livestock and poultry excrement, can also be used for regulating the water quality in culture water, regulating and controlling the structure of phytoplankton or used as a feed additive, can provide high-quality natural feed for zooplankton, and can also be used as high-quality feed for chironomid larvae for mass culture of the chironomid larvae.
In addition, the invention also relates to the application of the photosynthetic bacteria formed by the method in sewage purification treatment, which can accelerate the degradation of organic matters in sewage.
The invention also relates to the application of the photosynthetic bacteria formed by the method in the aspect of new energy, and the photosynthetic bacteria can be used for efficiently preparing hydrogen.
Has the advantages that:
compared with the prior art, the method for quickly and continuously culturing the photosynthetic bacteria of the rhodopseudomonas in the attached growth state has the following advantages:
1. the raw materials are easy to obtain, rich in source and low in cost;
2. the invention provides a simple and practical method for large-scale culture of photosynthetic bacteria, and the culture time is shorter than that of the conventional method and the efficiency is higher.
3. Compared with the photosynthetic bacteria growing in a floating state, the thick pulpy photosynthetic bacteria growing in an attached state on the inner wall of the container obtained by the invention can greatly reduce the concentration cost of the photosynthetic bacteria, save space for transportation and storage and be very convenient.
4. The culture method of the invention does not need measures such as stirring or aeration to promote the decay of the organic matters, thereby greatly saving the economic cost.
5. The high-concentration photosynthetic bacteria formed by the method can be directly used for treating breeding wastewater or livestock and poultry excrement, can also be used as high-quality natural bait for zooplankton, and can also be directly used for sewage purification treatment to efficiently prepare hydrogen.
Drawings
FIG. 1 is a photograph of a photosynthetic bacterium of the genus Rhodopseudomonas grown in an attached state cultured in example 1 of the present invention;
FIG. 2 is a photograph of a photosynthetic bacterium of the genus Rhodopseudomonas grown in an attached state cultured in example 2 of the present invention;
FIG. 3 is a photograph of a photosynthetic bacterium of the genus Rhodopseudomonas grown in an attached state cultured in example 3 of the present invention;
FIG. 4 is a photograph of a photosynthetic bacterium of the genus Rhodopseudomonas grown in an attached state cultured in example 4 of the present invention;
FIG. 5 is a photograph of an attached growth photosynthetic bacterium of the genus Rhodopseudomonas cultured in example 5 of the present invention.
Detailed Description
The present invention will be described in detail with reference to examples. The following examples will assist those skilled in the art in further understanding the invention, but are not intended to limit the invention in any way. It should be noted that it would be apparent to those skilled in the art that several modifications and improvements can be made without departing from the inventive concept. All falling within the scope of the present invention.
Example 1
At the bottom of 4 months, a colorless glass jar with the specification of 60cm multiplied by 40cm is placed in a glass greenhouse, 80L of water body containing phytoplankton cultured by tap water is added into the glass jar, the total nitrogen concentration in the water body is 2.0mg/L, the chlorophyll a concentration is 30 mu g/L, and green grass cut from a landscape grassland is added into the glass jar and is uniformly laid as much as possible, the adding thickness of the green grass in the glass jar is about 5cm, and the green grass floats on the upper layer of the water body.
The food waste was taken from the restaurant and after removing the non-food substances, the water was drained and spread out and air dried for 1 hour. 1200g of solid food waste was weighed, and 3g of monocalcium phosphate monohydrate [ Ca (H) ]was weighed2PO4)2·H2O]0.12g ferric citrate, 2 kinds of salts are mixed in the food waste and are evenly stirred and then tiled and added on the surface of the grass, the local stacking height is not more than 3cm, and the food waste is just submerged in water, so as to prevent mosquitoes from laying eggs on the food waste to generate mosquito larvae and other mosquito larvae, thereby reducing the utilization rate of the food waste.
Culturing for 3 days, and making the water body green and thick; culturing for 6 days, wherein the water color is changed from dark green to brown green, and green attached algae grow in the area of the upper part of the inner wall of the container, which is close to the grass; culturing for 11 days, and enlarging the area of the dark green attached algae on the inner wall of the container; on day 12, 4g magnesium hydrogen phosphate trihydrate (MgHPO) was added to the water under the grass4·3H2O)。
After 15 days of culture, the water body is still green and brown, dark red attachments appear in green attached algae in the area of the upper part of the inner wall of the container and close to the grass, and the dark green attached algae gradually turns into dark brown; after culturing for 19 days, the dark brown attachments on the inner wall of the container are completely changed into dark red, and high-throughput analysis and detection of microorganisms by using an Illumina MiSeq platform show that the dark red attached bacteria are Rhodopseudomonas sp (Rhodopseudomonas aeruginosa) photosynthetic bacteria, and the predominant species is Rhodopseudomonas putida (Rhodopseudomonas faecalis), namely the attached and growing Rhodopseudomonas sp photosynthetic bacteria are formed. As it continues to grow, the maximum thickness of the attached growing Rhodopseudomonas photosynthetic bacteria can reach about 1 mm.
The attached and growing photosynthetic bacteria of the genus Rhodopseudomonas can be scraped off by a cloth or other method without draining the water in the container, and then the same amount of food waste is continuously added to the grass in the container, wherein the same proportion of monocalcium phosphate monohydrate and ferric citrate is mixed in an amount not exceeding the amount of the previous addition, and magnesium hydrogen phosphate trihydrate is not added in the subsequent process. Because the water body also contains more rhodopseudomonas photosynthetic bacteria and other bacteria capable of degrading organic matters, the subsequent culture time is shorter, and the inner wall can grow with the attached rhodopseudomonas photosynthetic bacteria again within 5 days. Then the adhered rhodopseudomonas can be scraped off continuously, and the food waste mixed with salt can be added again for culture. Thus realizing continuous multiple culture until the grass therein is rotten and can not support the degradation of the food waste on the water surface.
In the time period from 4 months to 5 months, the room temperature of the glass greenhouse in the daytime and at night varies from 15 ℃ to 42 ℃. A photograph of the photosynthetic bacteria of the genus Rhodopseudomonas grown in the attached state as cultured in this example is shown in FIG. 1.
Example 2
At the end of 4 months, a 60cm × 35cm × 40cm colorless glass jar was placed in a glass greenhouse, and 70L of a phytoplankton-containing water cultured with tap water, in which the total nitrogen concentration was 30mg/L and the chlorophyll a concentration was 2700 μ g/L, was added to the glass jar, and which contains abundant phytoplankton of the genus Chlorophyceae. And green grass cut from the landscape grassland is added into the glass tank, the adding thickness of the green grass in the glass tank is about 7cm, and the green grass floats on the upper layer of the water body.
The food waste was taken from the restaurant and after removing the non-food substances, the water was drained and spread out and air dried for 1 hour. Weighing 4550g of solid food waste, 22.75g of monocalcium phosphate monohydrate and 1.14g of ferric citrate, mixing 2 salts into the food waste, uniformly stirring, spreading on the surface of green grass, wherein the local stacking height is not more than 3cm, so that the food waste is just submerged in water, and mosquitoes are prevented from laying eggs on the food waste to generate mosquito larvae and other mosquito larvae, thereby reducing the utilization rate of the food waste.
Culturing for 3 days until the water turns into brown green; culturing for 8 days, wherein the water color turns dark brown, and green attached algae grow in the area of the upper part of the inner wall of the container, which is close to the grass; culturing for 12 days, wherein the area of the dark green attached algae on the inner wall of the container is enlarged to about half of the area of the whole inner wall; on day 13, 15g of magnesium hydrogen phosphate trihydrate was added to the water below the grass.
After culturing for 18 days, dark red attachments appear in the green attached algae in the area of the upper part of the inner wall of the container, which is close to the grass, the dark green attached algae gradually turns into dark brown, and the water also turns into dark brown; after 22 days of culture, dark brown attachments on the inner wall of the container are fully changed into blood red, and high-throughput analysis and detection of microorganisms by using the Illumina MiSeq platform show that the blood red attachment bacteria are Rhodopseudomonas sp (Rhodopseudomonas aeruginosa) photosynthetic bacteria, and the predominant species is Rhodopseudomonas faecalis (Rhodopseudomonas faecalis), namely the attachment-growing Rhodopseudomonas sp photosynthetic bacteria are formed. As it continues to grow, the maximum thickness of the attached growing Rhodopseudomonas photosynthetic bacteria can reach about 2 mm.
The attached and growing photosynthetic bacteria of the genus Rhodopseudomonas can be scraped off by a cloth or other method without draining the water in the container, and then one third of the amount of food waste is continuously added to the grass in the container, wherein monocalcium phosphate monohydrate and ferric citrate are mixed in the same ratio, and magnesium hydrogen phosphate trihydrate is not added in the subsequent process. Because the water body also contains more rhodopseudomonas photosynthetic bacteria and other bacteria capable of degrading organic matters, the subsequent culture time is shorter, and the inner wall can grow with the attached rhodopseudomonas photosynthetic bacteria again in 8 days. Then the adhered rhodopseudomonas can be scraped off continuously, and the food waste mixed with salt can be added again for culture. Thus realizing continuous multiple culture until the grass therein is rotten and can not support the food waste to be degraded on the water surface.
In the period from 4 months bottom to 6 months early, the room temperature change range of the day and night in the glass greenhouse is 15-42 ℃. A photograph of the photosynthetic bacteria of the genus Rhodopseudomonas grown in the attached state as cultured in this example is shown in FIG. 2.
Example 3
At the end of 4 months, a 60cm × 35cm × 40cm colorless glass jar was placed in a glass greenhouse, and 70L of a phytoplankton-containing water cultured with tap water, in which the total nitrogen concentration was 20.3mg/L and the chlorophyll a concentration was 1260 μ g/L, was added to the glass jar, which contained abundant phytoplankton of the genus Chlorophyceae. And adding the grass cut from the landscape grassland into the glass cylinder, wherein the grass is uniformly paved as much as possible, the adding thickness of the grass in the glass cylinder is about 6cm, and the grass floats on the upper layer of the water body.
The food waste was taken from the restaurant and after removing the non-food substances, the water was drained and spread out and air dried for 1 hour. 3600g of solid food waste is weighed, 27g of monocalcium phosphate monohydrate, 27g of ferric citrate and 0.54g of ferric citrate are respectively weighed, salts are mixed in the food waste and uniformly stirred and then are tiled on the surface of green grass, the local stacking height is not more than 3cm, and the food waste is just submerged in water, so that mosquitoes can be prevented from laying eggs on the food waste to generate mosquito larvae and other mosquito larvae, and the utilization rate of the food waste is reduced.
Culturing for 3 days, wherein the water body is green and has a brown color; culturing for 9 days, wherein the color of the water body is changed from dark green to brown, and green attached algae grow in the area, close to the grass, on the upper part of the inner wall of the container; cultured for 12 days, deep on the inner wall of the vesselThe area of the green attached algae is enlarged to about half of the area of the whole inner wall; on day 14, 12g magnesium hydrogen phosphate trihydrate (MgHPO) was added to the water under the grass4·3H2O)。
After 18 days of culture, dark red attachments appear in the green attached algae in the area of the upper part of the inner wall of the container, which is close to the grass, and the dark green attached algae gradually turns into dark brown; after 22 days of culture, dark brown attachments on the inner wall of the container are completely changed into dark red, and high-throughput analysis and detection of microorganisms by using the Illumina MiSeq platform show that the dark red attachment bacteria are Rhodopseudomonas sp (Rhodopseudomonas aeruginosa) photosynthetic bacteria, and the dominant species is Rhodopseudomonas faecalis (Rhodopseudomonas faecalis), namely the attachment-growing Rhodopseudomonas sp photosynthetic bacteria are formed. As it continues to grow, the maximum thickness of the attached growing Rhodopseudomonas photosynthetic bacteria can reach about 2 mm.
The attached and growing photosynthetic bacteria of the genus Rhodopseudomonas can be scraped off by a cloth or other method without draining the water in the container, and then one third of the amount of food waste is continuously added to the grass in the container, wherein monocalcium phosphate monohydrate and ferric citrate are mixed in the same ratio, and magnesium hydrogen phosphate trihydrate is not added in the subsequent process. Because the water body also contains more rhodopseudomonas photosynthetic bacteria and other bacteria capable of degrading organic matters, the subsequent culture time is shorter, and the inner wall can grow with the attached rhodopseudomonas photosynthetic bacteria again in 8 days. Then the adhered rhodopseudomonas can be scraped off continuously, and the food waste mixed with salt can be added again for culture. Thus realizing continuous multiple culture until the grass therein is rotten and can not support the food waste to be degraded on the water surface.
In the time period from 4 months to 5 months, the room temperature of the glass greenhouse in the daytime and at night varies from 15 ℃ to 42 ℃. A photograph of the photosynthetic bacteria of the genus Rhodopseudomonas grown in the attached state as cultured in this example is shown in FIG. 3.
Example 4
At the end of 7 months, 10L of a colorless glass jar was placed in a glass greenhouse, and 10L of a phytoplankton-containing water cultured with tap water, in which the total nitrogen concentration was 24.6mg/L and the chlorophyll a concentration was 1178. mu.g/L, was added to the glass jar, which contained abundant phytoplankton of the genus Chlorophyceae. And adding the grass cut from the landscape grassland into the glass cylinder, wherein the grass is uniformly paved as much as possible, the adding thickness of the grass in the glass cylinder is about 5cm, and the grass floats on the upper layer of the water body.
The food waste was taken from the restaurant and after removing the non-food substances, the water was drained and spread out and air dried for 1 hour. Weigh 500g solid food and beverage rubbish, weigh 2g monocalcium phosphate monohydrate, 0.075g ferric citrate, mix salt and tile to the grass surface after the stirring in the food and beverage rubbish, and local pile height is no longer than 3cm, and food and beverage rubbish submerges just in the aquatic and is suitable to prevent that the mosquito from laying eggs on food and beverage rubbish and generating mosquito larvae etc. mosquito larva and reduce the utilization ratio of food and beverage rubbish.
Culturing for 3 days, and turning the water green to brown; culturing for 9 days, wherein the water color turns brown, and green attached algae grow in the area of the upper part of the inner wall of the container, which is close to the grass; culturing for 12 days, wherein the area of the dark green attached algae on the inner wall of the container is enlarged to about half of the whole area; on day 14, 4g of magnesium hydrogen phosphate trihydrate was added to the water below the grass.
After 18 days of culture, dark red attachments appear in the green attached algae in the area of the upper part of the inner wall of the container and close to the grass, and the dark green attached algae gradually turns into dark brown; after 22 days of culture, dark brown attachments on the inner wall of the container are fully changed into blood red, and high-throughput analysis and detection of microorganisms by using the Illumina MiSeq platform show that the blood red attachment bacteria are Rhodopseudomonas sp (Rhodopseudomonas aeruginosa) photosynthetic bacteria, and the predominant species is Rhodopseudomonas faecalis (Rhodopseudomonas faecalis), namely the attachment-growing Rhodopseudomonas sp photosynthetic bacteria are formed. As it continues to grow, the maximum thickness of the attached growing Rhodopseudomonas photosynthetic bacteria can reach about 2 mm.
The attached and growing photosynthetic bacteria of the genus Rhodopseudomonas can be scraped off by a cloth or other method without draining the water in the container, and then one third of the amount of food waste is continuously added to the grass in the container, wherein monocalcium phosphate monohydrate and ferric citrate are mixed in the same ratio, and magnesium hydrogen phosphate trihydrate is not added in the subsequent process. Because the water body also contains more rhodopseudomonas photosynthetic bacteria and other bacteria capable of degrading organic matters, the subsequent culture time is shorter, and the inner wall can grow with the attached rhodopseudomonas photosynthetic bacteria again in 8 days. Then the adhered rhodopseudomonas can be scraped off continuously, and the food waste mixed with salt can be added again for culture. Thus realizing continuous multiple culture until the grass therein is rotten and can not support the food waste to be degraded on the water surface.
In the period from the bottom of 7 months to the beginning of 9 months, the room temperature change range of the day and the night in the glass greenhouse is 20-45 ℃. A photograph of the photosynthetic bacteria of the genus Rhodopseudomonas grown in the attached state as cultured in this example is shown in FIG. 4.
Example 5
At the end of 9 months, a 60cm × 35cm × 40cm colorless glass jar was placed in a glass greenhouse, and 40L of a phytoplankton-containing water cultured with tap water, in which the total nitrogen concentration was 15.8mg/L and the chlorophyll a concentration was 565 μ g/L, was added to the glass jar, which contained abundant phytoplankton of the genus Chlorophyta and the genus Cellulosia. And adding the grass cut from the landscape grassland into the glass cylinder, wherein the grass is uniformly paved as much as possible, the adding thickness of the grass in the glass cylinder is about 4cm, and the grass floats on the upper layer of the water body.
The food waste was taken from the restaurant and after removing the non-food substances, the water was drained and spread out and air dried for 1 hour. Weigh 2000g solid food waste, weigh 6g monocalcium phosphate monohydrate, 0.3g ferric citrate, mix salt and tile to the grass surface after stirring in food waste, and local pile-up height is no longer than 3cm, and food waste submerges in the aquatic just suitably to prevent that the mosquito from laying eggs on food waste and generating mosquito larvae etc. mosquito larvae and reduce food waste's utilization ratio.
Culturing for 3 days, wherein the green color of the water body is light brown; culturing for 9 days, wherein the water color turns brown, and green attached algae grow in the area of the upper part of the inner wall of the container, which is close to the grass; culturing for 12 days, wherein the area of the dark green attached algae on the inner wall of the container is enlarged to about half of the whole area; on day 14, 6g of magnesium hydrogen phosphate trihydrate was added to the water below the grass.
After 18 days of culture, dark red attachments appear in the green attached algae in the area of the upper part of the inner wall of the container, which is close to the grass, and the dark green attached algae gradually turns into dark brown; after 20 days of culture, dark brown attachments on the inner wall of the container are completely changed into dark red, and high-throughput analysis and detection of microorganisms by using the Illumina MiSeq platform show that the dark red attachment bacteria are Rhodopseudomonas sp (Rhodopseudomonas aeruginosa) photosynthetic bacteria, and the dominant species is Rhodopseudomonas faecalis (Rhodopseudomonas faecalis), namely the attachment-growing Rhodopseudomonas sp photosynthetic bacteria are formed. As it continues to grow, the maximum thickness of the attached growing Rhodopseudomonas photosynthetic bacteria can reach about 2 mm.
The attached and growing photosynthetic bacteria of the genus Rhodopseudomonas can be scraped off by a cloth or other method without draining the water in the container, and then one third of the amount of food waste is continuously added to the grass in the container, wherein monocalcium phosphate monohydrate and ferric citrate are mixed in the same ratio, and magnesium hydrogen phosphate trihydrate is not added in the subsequent process. Because the water body also contains more rhodopseudomonas photosynthetic bacteria and other bacteria capable of degrading organic matters, the subsequent culture time is shorter, and the inner wall can grow with the attached rhodopseudomonas photosynthetic bacteria again in 8 days. Then the adhered rhodopseudomonas can be scraped off continuously, and the food waste mixed with salt can be added again for culture. Thus realizing continuous multiple culture until the grass therein is rotten and can not support the food waste to be degraded on the water surface.
In the time period from 9 months to 10 months, the room temperature change range of the day and the night in the glass greenhouse is 20-45 ℃. A photograph of the photosynthetic bacteria of the genus Rhodopseudomonas grown in the attached state as cultured in this example is shown in FIG. 5.
The structures, ratios, sizes, quantities, and the like shown in the drawings of the embodiments of the present invention are only used for matching with the disclosure of the specification, so as to be understood and read by those skilled in the art, and are not used to limit the conditions that the present invention can be implemented, so that the present invention has no technical significance, and any modification, ratio relationship change or size adjustment should still fall within the scope of the technical content of the present invention without affecting the efficacy and the achievable purpose of the present invention.
Claims (7)
1. A method for the rapid and continuous cultivation of photosynthetic bacteria of the genus Rhodopseudomonas with vegetative growth, comprising the steps of:
s1, placing the light-transmitting container in a glass greenhouse or an outdoor natural illumination environment, wherein the room temperature change range of day and night is 15-45 ℃, adding phytoplankton water into the light-transmitting container, and adding grass or straw rich in cellulose into the light-transmitting container to ensure that the grass or straw floats on the upper layer of the water;
s2, adding the solid food waste mixed with the salt mixture to grass or straws, wherein the amount of the grass or straws added with the rich cellulose in the step S1 is enough to prevent the solid food waste from sinking into water and just submerge the solid food waste in the water;
s3, culturing for 3-5 days, and enabling the water body to become dark green and even brown green;
culturing for 6-10 days, and growing green attached algae in the area where the upper part of the inner wall of the container is close to the grass;
culturing for 11-14 days, enlarging the area of the deep green attached algae on the upper part of the inner wall of the container, and adding magnesium hydrogen phosphate trihydrate (MgHPO) when the deep green attached algae grows4∙3H2O), the addition amount is 4-15 g;
s4, culturing for 15-18 days, wherein dark red attachments appear in green attached algae on the upper part of the inner wall of the container, and the dark green attached algae gradually turns into dark brown;
culturing for 19-25 days, wherein dark brown attachments on the inner wall of the container are completely changed into dark red or blood red rhodopseudomonas photosynthetic bacteria, and the area of the upper part of the inner wall of the container, which is close to the grass, is changed into dark green or continuously keeps dark red;
s5, scraping the rhodopseudomonas photosynthetic bacteria attached and grown in the step S4 by cloth under the condition that water in the container is not discharged; then continuously adding the food wastes mixed with salts on the grass or the straws in the container, wherein the adding amount is not more than that of the previous adding amount, and magnesium hydrogen phosphate trihydrate does not need to be added in the subsequent process, so that continuous multiple culture is realized until the grass or the straws are rotten and can not support the degradation of the food wastes on the water surface;
in the step S2, the adding proportion of the solid food waste to the water body is that the solid food waste is water body = 15-65 g:1L, the stacking thickness of the food waste on the grass or the straw is not more than 3cm, and the solid food waste is completely submerged by water;
in step S2, the salt mixture includes monocalcium phosphate monohydrate [ Ca (H) ]2PO4)2∙H2O]And ferric citrate, wherein the mass addition proportion of the monocalcium phosphate monohydrate is 0.25-0.75% of the mass of the solid food waste, and the mass addition proportion of the ferric citrate is 0.1-0.25% of the mass of the solid food waste.
2. A method of rapid and continuous culture of photosynthetic bacteria of the genus Rhodopseudomonas in adherent growth according to claim 1, wherein: the step S1 is performed in a warm season from late 4 th to early 11 th of month.
3. A method of rapid and continuous culture of photosynthetic bacteria of the genus Rhodopseudomonas in adherent growth according to claim 1, wherein: in step S2, the solid food waste is food waste left after removing non-food substances, and includes several or all of rice, wheaten food, fish, meat, eggs, vegetables, fruits, bones, and eggshells, and mainly contains rice and wheaten food starch substances, so as to ensure that liquid water in the food waste is as little as possible, and the food waste is spread out and air-dried for 1 hour for later use.
4. A method of rapid and continuous culture of photosynthetic bacteria of the genus Rhodopseudomonas in adherent growth according to claim 1, wherein: in the step S1, the transparent container is made of common colorless transparent glass, white transparent plastic or colorless transparent plastic.
5. A method of rapid and continuous culture of photosynthetic bacteria of the genus Rhodopseudomonas in adherent growth according to claim 1, wherein: in step S1, the grass or straw is straw or grass or wheat straw for greening lawn, and after being stacked to a certain thickness, the solid food waste can be kept on the water surface without sinking into the water bottom.
6. A method of rapid and continuous culture of photosynthetic bacteria of the genus Rhodopseudomonas in adherent growth according to claim 1, wherein: in the step S4, the maximum thickness of the attached and grown Rhodopseudomonas photosynthetic bacteria can reach 1-2 mm, and tap water is supplemented at any time during the culture period to supplement evaporation water consumption.
7. A method of rapid and continuous culture of photosynthetic bacteria of the genus Rhodopseudomonas in adherent growth according to claim 1, wherein: in the step S1, the total nitrogen concentration in the fresh water body containing the phytoplankton which is initially added is 2-30 mg/L, and the chlorophyll a concentration is 30-2700 mug/L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810694582.6A CN109280627B (en) | 2018-06-29 | 2018-06-29 | Method for rapidly and continuously culturing attached growth rhodopseudomonas photosynthetic bacteria |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810694582.6A CN109280627B (en) | 2018-06-29 | 2018-06-29 | Method for rapidly and continuously culturing attached growth rhodopseudomonas photosynthetic bacteria |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109280627A CN109280627A (en) | 2019-01-29 |
| CN109280627B true CN109280627B (en) | 2021-02-05 |
Family
ID=65185997
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810694582.6A Active CN109280627B (en) | 2018-06-29 | 2018-06-29 | Method for rapidly and continuously culturing attached growth rhodopseudomonas photosynthetic bacteria |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109280627B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105754903A (en) * | 2016-04-15 | 2016-07-13 | 中国水产科学研究院渔业机械仪器研究所 | Method for cultivating photosynthetic bacteria in Rhodopseudomonas on large scale |
| CN106282056A (en) * | 2016-08-12 | 2017-01-04 | 中国水产科学研究院渔业机械仪器研究所 | A kind of method utilizing food garbage to cultivate Rhodopseudomonas photosynthetic bacteria |
| CN107686815A (en) * | 2017-10-17 | 2018-02-13 | 中国水产科学研究院渔业机械仪器研究所 | A kind of method that chlorella is cultivated using food garbage |
| CN107858313A (en) * | 2017-12-12 | 2018-03-30 | 中国水产科学研究院渔业机械仪器研究所 | Culture medium and cultural method for Rhodopseudomonas photosynthetic bacteria apposition growth |
-
2018
- 2018-06-29 CN CN201810694582.6A patent/CN109280627B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105754903A (en) * | 2016-04-15 | 2016-07-13 | 中国水产科学研究院渔业机械仪器研究所 | Method for cultivating photosynthetic bacteria in Rhodopseudomonas on large scale |
| CN106282056A (en) * | 2016-08-12 | 2017-01-04 | 中国水产科学研究院渔业机械仪器研究所 | A kind of method utilizing food garbage to cultivate Rhodopseudomonas photosynthetic bacteria |
| CN107686815A (en) * | 2017-10-17 | 2018-02-13 | 中国水产科学研究院渔业机械仪器研究所 | A kind of method that chlorella is cultivated using food garbage |
| CN107858313A (en) * | 2017-12-12 | 2018-03-30 | 中国水产科学研究院渔业机械仪器研究所 | Culture medium and cultural method for Rhodopseudomonas photosynthetic bacteria apposition growth |
Non-Patent Citations (2)
| Title |
|---|
| Utilization of Alcohols by Rhodopseudomonas sp. No.7 Isolated from n-Propanol-Enrichment Cultures;Takaaki Finn et al.;《Agricultural and Biological Chemistry》;20140909;2747-2753 * |
| 一种光合细菌— 红假单胞菌分离物培养条件的研究;陈声明等;《浙江农业大学学报》;19951231;501-502 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109280627A (en) | 2019-01-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20190045756A1 (en) | Dark-bright integrated greenhouse system in intensive recirculating eco-aquaculture and aquaculture method | |
| Cahill et al. | Keeping the water clean—Seaweed biofiltration outperforms traditional bacterial biofilms in recirculating aquaculture | |
| JP4384141B2 (en) | How to use recycled water | |
| CN103250668A (en) | System combining aquaculture and soilless agriculture planting | |
| CN106277320A (en) | A kind of Penaeus vannamei freshwater cultivation water regulation method | |
| WO2008068829A1 (en) | Method for producing biomineral-containing substance and organic hydroponics method | |
| CN101591040A (en) | Method for preventing and controlling water bloom pollution organisms in eutrophicated water body | |
| CN105543148B (en) | A method of Rhodopseudomonas photosynthetic bacteria is cultivated using cyanobacterial bloom | |
| CN101823812A (en) | Method for treating pollution biogas slurry of poultry and livestock breeding | |
| CN108358692A (en) | It is a kind of to utilize liquid fertilizer of livestock and poultry feces and preparation method thereof and its application process | |
| CN105600942A (en) | Method for formation of bioflocs by cyanobacterial bloom | |
| JP5224799B2 (en) | Recycled water use method by remote control of organic wastewater treatment | |
| CN216452710U (en) | Brocade carp and vegetable symbiotic recirculating aquaculture system | |
| CN104839079B (en) | A kind of utilization prawn culturing waste water indoor scale cultivates the method and facility of Duo Lin Xi parent fishs | |
| CN103667109B (en) | A kind of Rhodobacter capsulatus and application thereof | |
| CN109280627B (en) | Method for rapidly and continuously culturing attached growth rhodopseudomonas photosynthetic bacteria | |
| CN106754385B (en) | Method for cultivating chlorella phytoplankton by using cyanobacterial bloom as raw material | |
| KR20110132674A (en) | Plant growing method using microorganism carrier and vegetable health drink thereof | |
| CN101781622A (en) | Vegetal bait culture method | |
| CN107858313A (en) | Culture medium and cultural method for Rhodopseudomonas photosynthetic bacteria apposition growth | |
| CN107686815B (en) | Method for cultivating chlorella by using food waste | |
| CN107667929B (en) | Cultivation method of deinsectization | |
| EP2473611B1 (en) | A system to produce feedstock for biogas production | |
| CN102812909B (en) | Method for purifying manure fermentation borth in dairy farm by loaches | |
| CN1580002A (en) | Method for preparing rich water compound bacteria and repairing aquaculture envionment |
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 |