Biological conditioner with water quality purification capacity as well as preparation method and application of biological conditioner

Abstract

The invention relates to the technical field of water treatment agents, in particular to a biological conditioner with water quality purification capacity and a preparation method and application thereof, and the biological conditioner is composed of a modified porous aluminosilicate inorganic material, an active microbial agent, fermentation liquor of the active microbial agent and an embedding material. The modified porous aluminosilicate inorganic material is diatomite, modified zeolite or perlite; the active microbial agent comprises bacillus, photosynthetic bacteria, alcaligenes faecalis, pseudomonas and nitrifying bacteria. The biological conditioner contains multiple beneficial microorganisms such as photosynthetic bacteria, nitrifying bacteria and bacillus, the microorganisms and multiple enzymes generated by the microorganisms have a synergistic effect, the respective purification effect can be achieved in a water body, harmful substances in the water body are synergistically degraded, and the self-purification capacity of the water body is improved.

Description

Technical Field

[0001] The present invention relates to the technical field of water treatment agents, and specifically relates to a biological conditioning agent with water purification capability, a preparation method and an application thereof. Background Art

[0002] In the aquaculture industry, water quality management is one of the key factors to ensure the healthy growth of aquaculture organisms and increase production. However, with the increase in aquaculture density and the extension of aquaculture cycles, a large amount of organic matter such as leftover bait, animal excrement, and plankton remains often accumulates in aquaculture waters. These substances decompose in water and produce harmful substances such as ammonia nitrogen and nitrite, causing water quality to deteriorate, which in turn affects the living environment and growth performance of aquaculture organisms.

[0003] When the nitrite content in the water is 0.1mg / L, the hemoglobin in the fish's blood turns into methemoglobin, turning the blood into chocolate color, which is commonly known as "brown blood disease" in aquaculture. It reduces the oxygen transport capacity of hemoglobin, causes gill swelling, reduces food intake, slows growth, and increases disease. When the concentration reaches 2.5mg / L, the fish is poisoned, respiratory function decreases, physical strength weakens, and finally breaks out and dies.

[0004] The lethal concentration of ammonia nitrogen to fish is 0.2-2.0 mg / L. When it is less than 0.2 mg / L, it will cause lesions, necrosis and tissue ulceration in the internal organs of fish. When the concentration reaches 0.01-0.02 mg / L, it is easy to damage the mucous membrane layer of the fish gills, reduce the ability of hemoglobin to carry oxygen, and slow the growth of fish. When the concentration reaches 0.02-0.05 mg / L, it will cause a variety of diseases and increase mortality. When the concentration reaches 0.05-0.2 mg / L, it will damage the skin, stomach, and intestinal mucosa of farmed fish, further causing bleeding in internal organs and body surface. When the concentration reaches 0.2-0.5 mg / L, it will cause acute poisoning and death of fish.

[0005] Traditional chemical water purifiers, such as polyaluminium chloride and polyferric sulfate, can flocculate harmful substances in the water through chemical reactions and sink them to the bottom of the pool, but this method often has the risk of secondary pollution and cannot fundamentally solve the water quality problem. Therefore, seeking a more environmentally friendly and efficient method for purifying aquaculture water has become an urgent need in the industry.

[0006] In recent years, with the development of microbiology and biotechnology, biological conditioners have shown great potential in the field of aquaculture water purification due to their non-toxic, easily degradable and eco-friendly characteristics. Biological conditioners mainly promote the degradation and mineralization of organic matter in water bodies through the growth, metabolism and extracellular enzyme production of microorganisms, thereby improving water quality. In addition, biological conditioners can also inhibit the growth of harmful microorganisms and promote the reproduction of beneficial microorganisms through niche competition, thereby rebuilding the ecological balance of aquaculture water bodies. Summary of the invention

[0007] The purpose of the present invention is to provide a biological conditioner with water purification ability, a preparation method and an application thereof, which has a stable treatment effect and can be used for long-term treatment without the need for additional equipment such as bacterial enrichment, bacterial cultivation, and film coating.

[0008] To achieve the above object, the present invention provides the following technical solutions:

[0009] A biological conditioner with water purification capability, comprising a modified porous aluminosilicate inorganic material, an active microbial agent and its fermentation liquid, and an embedding material;

[0010] The modified porous aluminosilicate inorganic material is diatomite, modified zeolite or perlite, which is not only a biological conditioner skeleton matrix, but also a microbial film filter bed, and can remove ammonia nitrogen and nitrite in water through cation exchange capacity and electrostatic adsorption capacity. The active microbial agent includes Bacillus, photosynthetic bacteria, Alcaligenes faecalis, Pseudomonas and nitrifying bacteria.

[0011] Wherein, the Bacillus is one or more of Bacillus thiamine-lyticus, Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus megaterium and Clostridium butyricum.

[0012] Wherein, the photosynthetic bacteria are one or more of Rhodopseudomonas palustris, Rhodospirillum rubrum and Rhodobacter sphaeroides.

[0013] Wherein, the Pseudomonas is Pseudomonas stutzeri.

[0014] Wherein, the nitrifying bacteria is one or more of Nitrosomonas, Nitrobacter, Nitrospinobacter gracilis, Nitrospira bacilli, and Nitrococcus.

[0015] The active microbial agent comprises 40-50 parts of photosynthetic bacteria, 20-30 parts of bacillus, 20-30 parts of nitrifying bacteria, 1-15 parts of alcaligenes faecalis, and 10-20 parts of pseudomonas.

[0016] Among them, according to the volume ratio or biomass ratio, photosynthetic bacteria: nitrifying bacteria: Bacillus: Pseudomonas: Alcaligenes faecalis = 4:2:2:1:1.

[0017] The above strains can be purchased from strain collection centers such as the China General Microbiology Center (CGMCC), China Center for Type Culture Collection (CCTCC) and the American Type Culture Collection (ATCC). For example:

[0018] The photosynthetic bacteria was Rhodopseudomonas palustris CGMCC 1.2180.

[0019] The nitrifying bacteria may be Nitrosospira Multiformis ATCC 25196; Nitrosomonas europaea ATCC 25978; or Nitrococcus mobilis ATCC25380.

[0020] The faecalis is Alcaligenes faecalis CGMCC1.924.

[0021] The bacillus may be Aneurinibacillus aneurinilyticus CGMCC1.16118, Bacillus amyloliquefaciens ATCC 23845, Bacillus Licheniformis CCTCC No. 2010437, or Clostridium butyricum CCTCC AB2010154.

[0022] The Pseudomonas was Pseudomonas stutzeri CGMCC 1.15316.

[0023] The number of live bacteria in the active microbial agent reaches 10 8 cfu / g or above.

[0024] Wherein, the embedding material is sodium alginate solidified by CaCl2 solution at 4°C.

[0025] The preparation method of the biological conditioning agent of the present invention comprises the following steps:

[0026] (1) Seed solution preparation: Under sterile conditions, Bacillus, photosynthetic bacteria, Alcaligenes faecalis, Pseudomonas and nitrifying bacteria are inoculated into seed culture medium respectively and cultured to a certain concentration;

[0027] (2) Fermentation culture: adding the seed liquid to a fermentation medium containing perlite or modified zeolite or porous ceramic particles, and continuing to culture until the number of bacteria reaches the required amount;

[0028] (3) Mixing preparation: The cultured microorganisms and the modified porous aluminosilicate inorganic material are mixed evenly in proportion and added to a certain proportion of sodium alginate solution. The mixed solution is dripped into a 4°C precooled CaCl2 solution using a peristaltic pump and cross-linked for 24 hours to obtain a biological conditioner.

[0029] The modified zeolite is selected as zeolite with a mesh size of 40-80 and modified with NaCl. The preparation method is as follows: the zeolite is placed in a 0.8 mol / L NaCl solution in a water bath at a constant temperature of 70-75°C for 3 hours, then taken out, washed and dried to obtain the NaCl modified zeolite.

[0030] Among them, the perlite is a particle with a particle size of 3-5 mm.

[0031] Among them, the preparation method of sodium alginate solution is: dissolving sodium alginate in water, and the sodium alginate powder quickly adheres together to form a sticky mass through hydration after meeting water. During the dissolution process, the stirring temperature needs to be controlled at about 80°C, stirring for 3 hours, and the rotation speed is 400rpm / min. It is slowly and completely hydrated and dissolved to form a viscous colloidal solution to ensure complete dissolution.

[0032] The crosslinking agent is a 4°C precooled CaCl2 solution. Water-soluble sodium alginate can be crosslinked by most multivalent cations (except magnesium and mercury) to form alginate gel. It is not limited to the listed solutions. The monovalent sodium ions in the sodium alginate G unit rapidly exchange ions with the divalent calcium ions in the solution at the carboxyl site, and then the calcium ions connect the carboxyl groups on the alginate molecular chains on both sides to form crosslinks. And the G unit can crosslink with Ca 2+ Complexation forms a unique egg-box structure (e.g. Figure 1 As shown in Figure 2, a calcium alginate gel structure with porous network space is finally formed. Calcium alginate is insoluble in water, can fix cells and has a certain mechanical strength.

[0033] The material is solidified and formed through cross-linking, and the microorganisms are fixed on the carrier material to form immobilized spheres. In this step, the dripping speed needs to be controlled by the peristaltic pump to ensure the quality and stability of the microbial agent after fixation. When mixing the microbial agent into the sodium alginate solution, it is necessary to pay attention to the temperature to be lowered to below 30°C to avoid inactivation of bacteria and enzymes in the microbial agent. However, it cannot be lower than 25°C. If the temperature is too low, the sodium alginate will easily solidify and clog the peristaltic pump pipeline.

[0034] Solidification and washing: The formed immobilized beads are further solidified in the solution, and then washed with physiological saline or other appropriate solutions to remove residual cross-linking agent and unfixed microorganisms.

[0035] Drying and storage: Dry the washed immobilized beads and then store them in a suitable container for future use. During the drying process, pay attention to the temperature and time control to avoid damage to the immobilized beads.

[0036] The biological conditioner prepared by the present invention can be used for water purification, especially for aquatic fish breeding water purification. Its mechanism of action is: using a porous carrier, a gel grid structure or a semi-permeable film to encapsulate Bacillus, photosynthetic bacteria, faecal alkaligenes, Pseudomonas and nitrifying bacteria cells, so that these bacteria can degrade small molecular substrates and products of ammonia nitrogen and nitrite and can enter and exit freely, while the cells will not diffuse into the surrounding medium to cause the loss of water in the breeding pond and the need for frequent feeding. The immobilized bacteria decompose organic matter and reduce harmful substances such as ammonia nitrogen and nitrite through metabolic activities.

[0037] Compared with the prior art, the present invention has the following beneficial effects:

[0038] The biological conditioner of the present invention contains a variety of beneficial microorganisms, such as photosynthetic bacteria, nitrifying bacteria, bacillus, etc. These microorganisms and the various enzymes produced by them work synergistically to play their own purification role in the water body, synergistically degrade harmful substances in the water body, and improve the self-purification ability of the water body. At the same time, the biological conditioner may also contain nutrients such as carbon sources, trace elements, vitamins, etc. to support the growth and metabolic activities of microorganisms.

[0039] All raw materials in the biological conditioner of the present invention are of biological or natural origin, are non-toxic and harmless, and will not cause secondary pollution to water bodies.

[0040] The present invention can prepare high-quality biological conditioning agents through steps such as solid culture medium culture, liquid culture medium culture, fermentation in a fermenter, filtration concentration, and preparation. It is not only easy to operate, but also has a good purification effect, is easy to industrialize, and is suitable for water purification projects of various scales, and does not require installation of equipment for use. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 Schematic diagram of the calcium alginate gel structure.

[0042] Figure 2 This is a curve of ammonia nitrogen content.

[0043] Figure 3 This is the change curve of nitrite content. DETAILED DESCRIPTION

[0044] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0045] Example 1

[0046] A biological conditioner with water purification capability is composed of modified porous aluminosilicate inorganic material, active microbial agent and fermentation liquid thereof, and embedding material.

[0047] The modified porous aluminosilicate inorganic material is diatomaceous earth.

[0048] Active microbial agents include Bacillus, photosynthetic bacteria, Alcaligenes faecalis, Pseudomonas and nitrifying bacteria.

[0049] Among them, the bacillus is selected from Bacillus amyloliquefaciens, the photosynthetic bacteria is selected from Rhodopseudomonas palustris, the pseudomonas is selected from Pseudomonas stutzeri, and the nitrifying bacteria is selected from Nitrosospirillum.

[0050] A single colony was picked from the plate and inoculated into 50 mL of sterilized nutrient broth medium and cultured in a shaking incubator at 30°C and 18 rpm for 14 h. 9 CFU / mL) culture solution was placed in a 50 mL centrifuge tube and centrifuged at 6000 r / min for 10 min. The supernatant was discarded, the bacterial precipitate was washed twice with sterile water, and then diluted to 10 mL with 0.9% saline to obtain a bacterial solution (bacterial concentration was 6.05×10 9 CFU / mL) for later use.

[0051] Weigh 2 g of sodium alginate and 2 g of diatomaceous earth, place them in 75 mL of deionized water, dissolve and sterilize in a water bath at 80 ° C, cool to about 30 ° C, add 25 mL of mixed bacterial solution (10 mL of Rhodopseudomonas palustris CGMCC1.2180, 5 mL of Bacillus amyloliquefaciens ATCC 23845, 5 mL of Nitrosospira Multiformis ATCC 25196, 2.5 mL of Alcaligenes faecalis CGMCC1.924, 2.5 mL of Pseudomonas stutzeri CGMCC 1.15316) and mix well to obtain 100 mL of 2% sodium alginate, 2% modified porous aluminosilicate inorganic material, and a bacterial density of 1.51×109 CFU / mL of solution.

[0052] The solution was dripped into a pre-cooled (4°C) 4% CaCl2 solution using a peristaltic pump, and the peristaltic pump speed was controlled at about 6-6.5 r / min. Different equipment can make corresponding adjustments (make sure that the pumped liquid does not form a line and drips out drop by drop without tailing). Crosslinked in a 4°C refrigerator for 24 hours, the obtained pellets were washed with sterile water and stored in 0.9% saline for later use.

[0053] Example 2

[0054] The bacteria mixed according to the proportion of Example 1 were cultured under shaking conditions at a speed of 180 r / min for 72 h at 30° C., and the fermentation supernatant was obtained after centrifugation at a speed of 10000 r / min for 10 min. The lipase activity was detected using a transparent circle of a tributyrin plate, that is, a hole was punched on the rescreened culture medium with a puncher with a diameter of 5 mm, 50 uL of the bacterial agent fermentation supernatant was injected into the hole, and cultured at 28° C. for 5 days. Whether there was a transparent circle around the culture solution was observed, and the enzyme activity was determined according to the size of the transparent circle.

[0055] Prepare skim milk powder solid culture medium, starch solid culture medium and cellulose solid culture medium respectively, inoculate single colonies on solid plates, invert and culture at 15°C for 48h, and observe the enzyme production type and transparent zone size.

[0056] It has been verified that the ratios of the transparent zone diameter to the colony diameter of the protease, amylase and cellulase in the fermentation broth of the microbial agent in Example 1 are 9.45±0.22, 5.37±0.15 and 3.35±0.17, respectively.

[0057] Example 3

[0058] Two fish tanks of 50×30×40 (cm), each tank contains 30 liters of water, and 20 grass goldfish are cultured. Each fish tank is not provided with a filtration system, and no oxygen is exposed. The two fish tanks are fed with equal amounts and times at regular intervals every day. The residual bait and excrement naturally settle and accumulate at the bottom of the fish tank. After one month of culture, the water body is turbid, and all indicators exceed the normal survival standard of fish. A blank control group and a microbial agent group (the biological conditioning agent of Example 1 is placed) are set up. The nitrite and ammonia nitrogen content values ​​in the water quality of the two fish tanks are monitored for 10 consecutive days.

[0059] After about 10 days of adding the biological conditioning agent prepared in Example 1, the ammonia nitrogen degradation rate reached 87.31%, and the nitrite degradation rate reached 99.49%. Figure 2-3 As shown in Table 1-2.

[0060] Table 1

[0061]

[0062] Table 2

[0063]

[0064] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A biological conditioner with water purification capability, characterized in that: It is composed of modified porous aluminosilicate inorganic material, active microbial agent and fermentation liquid thereof, and embedding material; Wherein, the modified porous aluminosilicate inorganic material is diatomaceous earth, modified zeolite or perlite; the active microbial agent includes Bacillus, photosynthetic bacteria, Alcaligenes faecalis, Pseudomonas and nitrifying bacteria.

2. The biological conditioner having water purification capability according to claim 1, characterized in that: The bacillus is one or more of Bacillus thiamine-lyticus, Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus megaterium and Clostridium butyricum.

3. The biological conditioner with water purification ability according to claim 1, characterized in that: The photosynthetic bacteria are one or more of Rhodopseudomonas palustris, Rhodospirillum rubrum and Rhodobacter sphaeroides.

4. The biological conditioner having water purification capability according to claim 1, characterized in that: The Pseudomonas is Pseudomonas stutzeri.

5. The biological conditioner having water purification capability according to claim 1, characterized in that: The nitrifying bacteria are one or more of Nitrosomonas, Nitrobacter, Nitrospinobacter gracilis, Nitrospira bacilli, and Nitrococcus.

6. The biological conditioner with water purification ability according to claim 1, characterized in that: The active microbial agent comprises 40-50 parts of photosynthetic bacteria, 20-30 parts of bacillus, 20-30 parts of nitrifying bacteria, 1-15 parts of alcaligenes faecalis, and 10-20 parts of pseudomonas.

7. The biological conditioner having water purification capability according to claim 6, characterized in that: According to the volume ratio or biomass ratio, photosynthetic bacteria: nitrifying bacteria: Bacillus: Pseudomonas: Alcaligenes faecalis = 4:2:2:1:1; the number of live bacteria in the active microbial agent reaches 10 8 cfu / g or above.

8. The biological conditioner with water purification ability according to claim 1, characterized in that: The embedding material is sodium alginate solidified by a CaCl2 solution at 4°C.

9. The method for preparing the biological conditioning agent according to any one of claims 1 to 8, characterized in that: The following steps are involved: (1) Seed solution preparation: under sterile conditions, Bacillus, photosynthetic bacteria, Alcaligenes faecalis, Pseudomonas and nitrifying bacteria are inoculated into seed culture medium respectively and cultured to a certain concentration; (2) Fermentation culture: adding the seed solution to the fermentation medium containing the modified porous aluminosilicate inorganic material, and continuing to culture until the number of bacteria reaches the required amount; (3) Mixing preparation: The cultured microorganisms and the modified porous aluminosilicate inorganic material are mixed evenly in proportion and added to a certain proportion of sodium alginate solution. The mixed solution is dripped into a 4°C precooled CaCl2 solution using a peristaltic pump and cross-linked for 24 hours to obtain a biological conditioner.

10. Use of the biological conditioner according to any one of claims 1 to 8 in water purification.

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