CN109607789B - Ecological bed of high-efficient nitrogen cycle bacterium - Google Patents
Ecological bed of high-efficient nitrogen cycle bacterium Download PDFInfo
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- CN109607789B CN109607789B CN201811642193.5A CN201811642193A CN109607789B CN 109607789 B CN109607789 B CN 109607789B CN 201811642193 A CN201811642193 A CN 201811642193A CN 109607789 B CN109607789 B CN 109607789B
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/30—Aerobic and anaerobic processes
- C02F3/302—Nitrification and denitrification treatment
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2203/00—Apparatus and plants for the biological treatment of water, waste water or sewage
- C02F2203/006—Apparatus and plants for the biological treatment of water, waste water or sewage details of construction, e.g. specially adapted seals, modules, connections
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Abstract
The invention belongs to the field of ecological protection, and a stable reaction area is formed in an ecological bed through the ecological bed of high-efficiency nitrogen circulating bacteria, so that the defect that the prior art is basically invalid or is forced to stop in autumn and winter is overcome, and the continuous operation of ecological engineering at a lower temperature is realized. Through the action of nitrifying bacteria and denitrifying bacteria, nitrogen in various forms in natural water, domestic sewage and industrial waste water is gradually removed according to the nitrogen circulation reaction, and the effect of removing nitrogen is obvious.
Description
Technical Field
The invention belongs to the technical field of environmental ecological engineering, relates to artificial screening of microorganisms and application thereof in environmental management, and particularly relates to a high-efficiency nitrogen circulation bacteria ecological bed.
Background
The pollution and eutrophication of water bodies such as lakes, rivers and the like are serious ecological environmental problems which endanger the health of human beings and restrict the sustainable development of socioeconomic property in China and many areas around the world. The existing researches show that nitrogen and phosphorus are main limiting factors in various nutrient substances causing water eutrophication. Therefore, one of the keys to solving the problem of water eutrophication is to control the nitrogen content of the water.
At present, the most research and application for domestic sewage and industrial wastewater treatment at home and abroad is a biological denitrification method, but the nitrification-denitrification process is difficult to unify in time and space, the denitrification effect is poor, and the multi-step biological catalytic reaction of biological denitrification is limited by substrate transfer rate, substrate and product inhibition and the like. Nitrification requires nitrifying bacteria, wherein nitrifying bacteria comprise nitrite bacteria and nitrate bacteria, nitrification can be completed only through the combined action of the nitrite bacteria and the nitrate bacteria, single bacteria cannot be completed, but the nitrification is reduced when the nitrite bacteria and the nitrate bacteria are in different environments, so that the nitrite bacteria and the nitrate bacteria need to be fused into a whole, and the nitrifying bacteria suitable for various environments can be screened.
Disclosure of Invention
The invention aims to solve the technical problem of providing an efficient nitrogen circulating bacteria artificial screening method aiming at the defects in the prior art.
In order to solve the technical problems, the invention adopts the technical scheme that: a high-efficiency nitrogen-circulating bacteria ecological bed comprises a high-efficiency nitrogen-circulating bacteria releaser, an ecological floating island and a fixing device for fixing the ecological floating island, wherein the ecological floating island comprises a plurality of PVC rods, the edges of the PVC rods are provided with outward-protruding buckles and inward-recessed locking grooves, a circular hole is formed in the center of one end of each PVC rod, a boss is arranged on the surface of the other end of each PVC rod, a regular polygonal pipe penetrating through the circular hole is arranged in the center of the boss, the regular polygonal pipe is positioned at one end of the boss and is open, the other end of the boss is closed, the high-efficiency nitrogen-circulating bacteria releaser is fixedly arranged at the open end of the regular polygonal pipe, the circular hole is fixedly connected with the regular polygonal pipe through a plurality of connecting plates, the connecting plates and the regular polygonal pipe divide the inside of the circular hole into a plurality of water outlet areas, a plurality of water through holes are formed in the regular polygonal pipe, water in the regular polygonal pipe flows into the plurality of water outlet areas through the plurality of water through holes, and flows out from the other end.
Preferably, the male buckle and the female lock groove are adjacently arranged.
Preferably, the male buckle and the female lock groove are symmetrically arranged.
Preferably, the boss is shaped as a regular hexagon.
Preferably, the boss is circular in shape.
Preferably, the fixing device comprises sliding grooves arranged on two sides of the river channel and a plurality of cross beams arranged above the river channel in a crossing manner, the cross beams move along the sliding grooves, and the ecological floating island is suspended on two adjacent cross beams.
Preferably, the ecological floating island four corners fixedly connected with jib, the jib be height-adjustable's screw rod, the upper end of jib is provided with the gyro wheel, the gyro wheel sets up on the crossbeam, and follow the crossbeam removes, jib upper end be provided with and pull the crossbeam.
The invention has the beneficial effects that: according to the invention, firstly, an ecological bed of efficient nitrogen circulating bacteria is reformed, a water inlet area is set into a wide hexagonal pipe, so that oxygen aeration is ensured, the aerobic nitrification reaction is better adapted, water flows into a narrow water outlet area through water through holes in the hexagonal pipe to form an anaerobic area, the denitrification reaction of the bacteria is ensured, and nitrogen in river water is fully decomposed; the invention also obtains the high-efficiency nitrogen circulating bacteria with better nitrogen removal effect by improving the screening and enrichment method of the strains. When the high-efficiency nitrogen circulating bacteria are applied to the treatment of domestic sewage and industrial wastewater and the ecological restoration of eutrophic water, the tolerance of the ecological engineering-immobilization technology to severe environment is enhanced, the defect that the prior art basically fails or is forced to stop in autumn and winter is overcome, and the continuous operation of the ecological engineering at lower temperature is realized
Drawings
FIG. 1 is a schematic structural view of the present invention;
FIG. 2 is a schematic view of the fixing device of the present invention;
fig. 3 is a top view of an ecological floating island in river water according to an embodiment of the present invention;
FIG. 4 is a schematic view of an end structure of a PVC rod according to an embodiment of the present invention;
FIG. 5 is a schematic view of another end of the PVC rod according to an embodiment of the present invention;
fig. 6 is a top view of an ecological floating island in river water according to another embodiment of the present invention;
FIG. 7 is a schematic view of an end structure of a PVC rod according to another embodiment of the present invention;
FIG. 8 is a schematic view showing another end structure of a PVC rod according to another embodiment of the present invention.
Description of reference numerals:
1-ecological floating island; 2-a fixing device; 3, a chute; 4, a cross beam; 5, rolling wheels; 6, a suspender; 7-pulling the beam 7; 8-PVC rod; 9-high nitrogen circulating bacteria releaser; 81-convex buckle; 82-inner concave locking groove; 83-circular hole; 84-regular polygon tube; 85, connecting plates; 86-a water outlet zone; 87-water hole; 88-a boss; 81 a-convex buckle; 82 a-inner concave locking groove; 83 a-circular hole; 84 a-regular polygon tube; 85 a-connecting plate; 86 a-water outlet zone; 87 a-water passage hole; 88 a-boss.
Detailed Description
The present invention is further described in detail below with reference to examples so that those skilled in the art can practice the invention with reference to the description.
It will be understood that terms such as "having," "including," and "comprising," as used herein, do not preclude the presence or addition of one or more other elements or groups thereof.
As shown in fig. 1-2, the embodiment provides an ecological bed of high-efficient nitrogen circulation bacterium, including high-efficient nitrogen circulation bacterium releaser 9, ecological chinampa 1 and the fixing device 2 of fixed ecological chinampa, fixing device 2 is including setting up spout 3 on river course both sides, span a plurality of crossbeams 4 that set up in the river course top, crossbeam 4 removes along spout 3, hangs on two adjacent crossbeams 4 and is equipped with ecological chinampa 1, ecological chinampa 1 four corners fixedly connected with jib 6, jib 6 is height-adjustable's screw rod, the upper end of jib 6 is provided with gyro wheel 5, gyro wheel 5 sets up on crossbeam 4, and removes along crossbeam 4, jib 6 upper end is provided with drags and draws crossbeam 7, and the design can be as required put multiunit ecological chinampa 1 in the river course, and jib 6 can change the position in the river, also conveniently gets and puts.
As shown in fig. 3-8, the ecological floating island 1 comprises a plurality of PVC rods 8, the edges of the PVC rods 8 are provided with outward convex buckles 81 and inward concave locking grooves 82, the center of one end of the PVC rod 8 is provided with a circular hole 83, the surface of the other end of the PVC rod 8 is provided with a boss 88, the center of the boss 88 is provided with a regular polygonal tube 84 penetrating through the circular hole 83, the regular polygonal tube 84 is positioned at one end of the boss 88 and is open, the other end of the boss 88 is closed, the high-efficiency nitrogen circulation bacteria releaser 9 is fixedly arranged at the open end of the regular polygonal tube 84, the circular hole 83 is fixedly connected with the regular polygonal tube 84 through a plurality of connecting plates 85, the connecting plates 85 and the regular polygonal tube 84 equally divide the inside of the circular hole 83 into a plurality of water outlet areas 86, the regular polygonal tube 84 is provided with a plurality of water through holes 87, the water in the regular polygonal tube 84 flows into the plurality of water outlet areas 86 through the plurality of water through holes 87, and is designed such that the high-efficiency nitrogen circulation bacteria releaser 9 is arranged at the water inlet of the PVC rod 8, through the river, release the bacterium in the PVC stick 8, the bacterium is attached to in the PVC stick 8 like this, can form stable good oxygen district and anaerobic zone, fully decomposes the nitrogen in the aquatic, realizes the river maximize and purifies.
Example 1
The convex buckle 81 and the concave lock groove 82 are adjacently arranged and are matched in shape, the direction of the PVC rod 8 is not required to be distinguished in the design, the ecological floating island 1 can be expectedly spliced quickly, the shape of the boss 88 is a regular hexagon, the cross section of the PVC rod 8 is the same, the procedure is not required to be replaced in the processing, the processing is convenient, and the boss 88 is additionally arranged to distinguish the direction of putting into water.
Example 2
The outer convex buckle 81a and the inner concave lock groove 82a are symmetrically arranged and are adaptive in shape, the spliced ecological floating island 1 is spliced along one direction due to the design, when the PVC rod 8 is damaged, the damaged part can be found out quickly along the splicing direction, the boss 88a is circular in shape, edges and corners do not exist in the circular design, pollutants in water cannot be hung easily, and the water inlet cannot be blocked.
As shown in fig. 3 to 6, the regular polygonal tubes are regular hexagonal tubes, but the regular polygonal tubes may be not only regular hexagonal tubes but also all other regular polygonal tubes.
The water inlet of regular hexagonal pipe 84 is equipped with high-efficient nitrogen cycle bacterium release, and the artifical screening step of high-efficient nitrogen cycle bacterium:
collecting a bacterial water sample:
a sterilized sterile test tube (1 multiplied by 1.5cm) is used for extending into a water layer of 10-20 cm below the water surface in water bodies such as lakes and the like for sampling, and the water layer is immediately sealed after sampling and is ready to be inoculated in a laboratory.
Culture medium:
nitrous oxideCulture medium of acid bacteria: (NH)4)2SO4 2.0g，NaHPO4 0.25g，MnSO4·4H2O0.01g，K2HPO40.75g，MgSO4·7H20.03g of O, 31.0 g of CaCO and 1000ml of distilled water, and sterilizing at the pH of 7.2121 ℃ for 30 min.
Culture medium of nitrate bacteria: NaNO2 1.0g，Na2CO3 1.0g,NaHPO4 0.25g，CaCO3 1.0g，K2HPO40.75g，MnSO4 0.01g,MgSO4·4H2O0.03 g, distilled water 1000ml, pH 7.2121 deg.C sterilization for 30 min.
Denitrifying bacteria culture medium:
KNO3 2.0g，MgSO4 0.2g，KHSO4 0.5g，NaKC4H4O6·4H2o, 1000ml of distilled water, and sterilizing at the pH of 7.2121 ℃ for 30 min.
The separation method comprises the following steps:
(1) enrichment culture
The collected sample was inoculated into a previously sterilized triangular flask containing glass beads and 90ml of a medium, and cultured at 17 ℃ for 10 days on a shaker at 150 r/min. Aims to break up zoogloea, enable bacteria to be dispersed in water in a single cell state, and increase the quantity of nitrifying bacteria in a mud sample.
(2) Plate dilution separation
Taking out 1mL of bacterial suspension from the enrichment culture bacterial suspension, and inoculating the bacterial suspension into a test tube filled with 9mL of sterile water to obtain 10-1Gradient bacterial suspension, which can be obtained by the method described above to obtain 10-2、10-3、10-4… gradient of bacterial suspension, selecting gradient 10-4—10-8Taking 1mL of the bacterial suspension, putting the bacterial suspension into a plate, adding a proper culture medium, uniformly mixing, inverting the plate after the culture medium is solidified, and putting the plate into an incubator at 17 ℃ for culturing for 7-14 days. After bacterial single cells or the same type of cells form colonies on a culture medium, taking the single colonies to culture on a slant culture medium, and marking for later use.
(3) Plate scribing and separating
And (4) picking a small amount of lawn of the slant culture medium of the strain after dilution separation, and carrying out plate streaking separation for further purification. The whole process requires aseptic operation.
First, a small number of growing bacteria were picked from the slant medium and then streaked onto a previously prepared plate. And during scribing, the scribed range is drawn to be full as much as possible, then the inoculating loop is burned, rotated by a certain angle to be connected with the scribed area, another area is drawn, finally the whole plate is drawn to be full, and the whole plate is placed into a 17 ℃ incubator to be cultured for 7-14d, so that a single colony is obtained. Different bacterial colonies are transferred to the inclined plane, and after repeated several times, a single bacterial strain can be obtained and transferred to the inclined plane for later experiments.
(4) Result of separation
According to the above method, 42 pure strains were isolated from the nitrite bacteria culture medium, 45 pure strains were isolated from the nitrate bacteria culture medium, and 27 pure strains were isolated from the denitrifying bacteria culture medium.
Protoplast fusion of nitrite and nitrate bacteria
1. Nitrite bacteria with selectable genetic markers are used as parents.
2. The protoplast is prepared, the cell wall is removed by lysozyme, the protoplast is released, and the protoplast is placed in high-penetrating fluid to maintain the stability of the protoplast.
3. The fusion-promoting polyethylene glycol is added into the protoplast to promote fusion, the polyethylene glycol is a surfactant and can forcibly promote the fusion of the protoplast, and the fusion can be promoted in the presence of Ca2+ and Mg2+ ions.
4. Protoplast regeneration protoplasts lose cell walls and, although biologically active, do not grow on normal media and must be plated on regeneration media to regenerate them.
5. Detection of the fusions utilizes genetic markers on the selection medium to determine whether they are fusions.
6. The fusants produced by the fusant screening may have different types of heterozygous diploid and haploid recombinants, the former has unstable performance, the haploid recombinants with stable performance are selected, and the fusants with good production performance are repeatedly screened.
The method comprises the following steps of applying high-efficiency immobilized nitrogen circulating bacteria (ammonification-nitrification-nitrosation-denitrification) to flowing water bodies such as rivers and the like:
(h) enriching the high-efficiency nitrogen circulating bacteria obtained by artificially screening the high-efficiency nitrogen circulating bacteria according to claim 1;
(j) mixing the enriched fusion bacteria and denitrifying bacteria optimization bacteria, and putting the mixture into a prepared carrier for immobilization;
(k) and (3) putting the prepared immobilized efficient nitrogen circulating bacteria into a wastewater treatment device for wastewater treatment or putting the immobilized efficient nitrogen circulating bacteria into a natural water body for ecological restoration.
In the step (j), an ecological floating island can be arranged in the natural water body, the ecological floating island is hung with an immobilized high-nitrogen circulating bacteria releaser, and the releaser places the immobilized nitrogen circulating bacteria carrier in a container with meshes, so that the immobilized high-efficiency circulating bacteria can contact the water body, and the immobilized nitrogen circulating bacteria is released into the natural water body in this way to carry out ecological restoration on the natural water body.
The specific implementation scheme is as follows:
A. taking a water sample in a natural water body aquatic plant growth root area to be inoculated
B. And (3) obtaining high-efficiency fusion bacteria and denitrifying bacteria of the nitrate bacteria and the nitrite bacteria by manual screening.
C. Mixing the two kinds of nitrogen circulating bacteria and putting the mixture into a carrier to prepare the high-efficiency immobilized nitrogen circulating bacteria.
D. And (3) surveying the surrounding natural environment and pollution sources of the water body, determining the flow state and inflow water quality of the adjacent water body, determining the specific distribution of the engineering area, and starting to monitor the water quality of the engineering area.
E. An ecological engineering experiment area is established in a water body and is divided into a control area, an immobilized nitrogen circulating bacteria experiment area and an aquatic plant + immobilized nitrogen circulating bacteria experiment area, 20kg of immobilized nitrogen circulating bacteria are divided into four parts of 5kg, 5kg, 5kg and 5kg, the four parts are respectively arranged at two ends of the immobilized nitrogen circulating bacteria experiment area and the aquatic plant + immobilized nitrogen circulating bacteria experiment area by ecological floating islands 1, each ecological floating island 1 is hung with an immobilized nitrogen circulating bacteria releaser, the releaser places immobilized nitrogen circulating bacteria carriers in a special container with meshes and is placed 20-30 cm below the water surface, and the immobilized nitrogen circulating bacteria can contact the water body.
F. The release of the nitrogen circulating bacteria in the carrier increases the number of the nitrogen circulating bacteria in the water body per unit volume, preliminarily increases a nitrification-denitrification interface in the water body, enhances the nitrogen release activity of the water body in an ecological engineering test area, and creates conditions for the next step of introducing aquatic higher plants.
G. And monitoring the water quality of the water in the engineering area, and introducing the aquatic high plants to the artificial floating island when the aquatic high plants are ensured to grow. The introduction of aquatic higher plants increases a plurality of rhizosphere micro-ecological systems in the water body and strengthens the treatment effect of nitrogen circulating bacteria. The release of the immobilized nitrogen circulating bacteria obviously increases the rhizosphere bacteria of the aquatic higher plants; and along with the movement of water flow, the nitrogen circulating bacteria further diffuse to the downstream, thereby promoting the nitrogen diffusion of the downstream water body and enhancing the self-cleaning capability of the whole watershed.
The application effect is as follows:
the method is applied to release nitrogen circulating bacteria, so that the number of bacteria in the root zone of the aquatic plants in the test zone is obviously influenced, the number of denitrifying bacteria and nitrifying bacteria (fusion of nitrate bacteria and nitrite bacteria) in the root zone of the aquatic plants in the ecological floating island 1 is between 4 and 6 orders of magnitude, is obviously higher than the number of bacteria in the root zone of the aquatic plants in the ecological floating island 1, and is averagely 1 to 2 orders of magnitude, wherein the MPN values of the denitrifying bacteria and the nitrifying bacteria reach an extremely obvious level (P is less than 0.01); f test results show that the MPN values of denitrifying bacteria at the water outlet are all remarkably higher than the MPN value at the water inlet (P is less than 0.01); the analysis of variance shows that the upper open water area, the lower open water area and the positions 10cm and 30cm away from the immobilized nitrogen circulating bacteria; the number of released nitrifying bacteria is obviously higher than that of water inlet positions at all positions in the test area, wherein the number of the nitrifying bacteria at the position 10cm away from the artificial floating island of the immobilized azotobacter is the highest, and the MPN value of the nitrifying bacteria is extremely higher than that of the water inlet positions (P is less than 0.001). During the test, the number of circulating bacteria in the downstream water zone was also significantly higher than at the intake.
After the immobilized nitrogen circulating bacteria is treated, the total nitrogen, nitrite nitrogen and nonionic ammonia in an experimental area are respectively obviously reduced compared with those before engineering, and TN, NO 3-and NH3 are respectively reduced to 0.21mg/L, 0.008mg/L and 0.05mg/L after the engineering from the highest values of 4.613mg/L, 0.075mg/L and 0.15mg/L before the engineering, which shows that the technology of adopting the high-efficiency immobilized nitrogen circulating bacteria has obvious effect on removing nitrogen in flowing water bodies such as rivers and the like.
In the description of the present invention, it is to be understood that the terms "central," "longitudinal," "lateral," "length," "width," "thickness," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," "clockwise," "counterclockwise," "axial," "radial," "circumferential," and the like are used in the orientations and positional relationships indicated in the drawings for convenience in describing the invention and to simplify the description, and are not intended to indicate or imply that the referenced devices or elements must have a particular orientation, be constructed and operated in a particular orientation, and are therefore not to be considered limiting of the invention.
While embodiments of the invention have been disclosed above, it is not limited to the applications listed in the description and the embodiments, which are fully applicable in all kinds of fields of application of the invention, and further modifications may readily be effected by those skilled in the art, so that the invention is not limited to the specific details without departing from the general concept defined by the claims and the scope of equivalents.
Claims (5)
1. A high-efficiency nitrogen-circulating bacteria ecological bed comprises a high-efficiency nitrogen-circulating bacteria releaser, an ecological floating island and a fixing device for fixing the ecological floating island, and is characterized in that the ecological floating island comprises a plurality of PVC rods, the edges of the PVC rods are provided with an outward-protruding buckle and an inward-recessed locking groove, a circular hole is formed in the center of one end of each PVC rod, a boss is arranged on the surface of the other end of each PVC rod, a regular-polygonal tube penetrating through the circular hole is arranged in the center of the boss, one end of each regular-polygonal tube is open, the other end of each regular-polygonal tube is closed, the high-efficiency nitrogen-circulating bacteria releaser is fixedly arranged at the open end of each regular-polygonal tube, the circular hole is fixedly connected with the regular-polygonal tubes through a plurality of connecting plates, the connecting plates and the regular-polygonal tubes divide the inside of the circular hole into a plurality of water outlet areas, and a plurality of water through holes are arranged on the regular-polygonal tubes, the water in the regular polygonal pipe flows into the water outlet areas through the water through holes and flows out from one end of the PVC rod with the round hole; the boss is in the shape of a regular hexagon.
2. The efficient nitrogen-circulating bacteria ecological bed as claimed in claim 1, wherein the male locking buckle and the female locking groove are disposed adjacently.
3. The efficient nitrogen-circulating bacteria ecological bed as claimed in claim 1, wherein the male locking buckle and the female locking groove are symmetrically arranged.
4. The efficient nitrogen-circulating bacteria ecological bed as claimed in claim 1, wherein the fixing device comprises chutes arranged at both sides of the river channel, and a plurality of beams arranged across and above the river channel, the beams moving along the chutes, and the ecological floating island being suspended on two adjacent beams.
5. The efficient nitrogen-circulating bacteria ecological bed as claimed in claim 4, wherein hanging rods are fixedly connected to four corners of the ecological floating island, the hanging rods are height-adjustable screws, rollers are arranged at upper ends of the hanging rods, the rollers are arranged on the cross beams and move along the cross beams, and dragging cross beams are arranged at upper ends of the hanging rods.
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