Preparation method and application of multi-strain-ceramsite carrier compound for static denitrification purification of aquaculture sewage

Abstract

The invention belongs to the technical field of wastewater and sewage treatment, and relates to a preparation method and application of a multi-strain-ceramsite carrier compound for static denitrification purification of aquaculture sewage. The multi-strain-ceramsite carrier compound consists of rhodopseudomonas palustris, pediococcus pentosaceus, bacillus licheniformis, candida albicans, leuconostoc mesenteroides, cellulomonas flavigena and staphylococcus saprophyticus, and is prepared by loading the multi-strain-ceramsite carrier compound on a ceramsite carrier through a special process. When the compound is used for treating aquaculture sewage, strain layered feeding is realized through ceramsite carrier density difference, the aquaculture sewage can be efficiently treated, and odor generation can be remarkably inhibited. Application results show that the total nitrogen degradation rate, the ammonia nitrogen degradation rate and the COD degradation rate of the culture sewage treated by the method are 86.2%, 88.2% and 79.2% respectively. Through the synergistic effect of multiple strains, process innovation and precise spatial distribution control, efficient static purification of the breeding sewage is achieved, and an economical and feasible technical scheme is provided for rural pollution treatment.

Description

Technical Field

[0001] This invention belongs to the field of wastewater and sewage treatment technology, specifically relating to a method for preparing a multi-strain ceramsite carrier composite for denitrification and purification of aquaculture sewage and its application. Background Technology

[0002] China's rural livestock industry has gradually shifted from a traditional individual household model to large-scale development, leading to further improvements in breeding techniques. However, the development of livestock farming has also generated a large amount of livestock and poultry wastewater, causing a series of environmental pollution problems. Statistics show that livestock and poultry wastewater is a major source of non-point source pollution in rural areas. This wastewater contains large amounts of livestock and poultry manure, urine, and undigested feed. The main components of feed include organic components such as protein, starch, oil, and cellulose. If this complex organic wastewater is discharged directly without effective treatment, it will lead to eutrophication and water quality deterioration.

[0004] Currently, wastewater treatment methods for aquaculture mainly include physical, chemical, land treatment, and biological methods. Among these, physical methods can only remove suspended solids to a limited extent and are difficult to degrade dissolved pollutants; chemical methods are costly and prone to secondary pollution; land treatment is slow to show results and may lead to heavy metal accumulation, threatening soil safety; biological treatment utilizes microorganisms (aerobic, anaerobic, and facultative anaerobic bacteria) to degrade pollutants, offering advantages such as large treatment capacity, low cost, and no secondary pollution, and has become a research hotspot. However, the effectiveness of microbial treatment is easily affected by environmental factors such as pH and temperature; if the strain combination is unreasonable or the adaptability is poor, it may lead to low nitrogen removal efficiency. Furthermore, rural mountainous areas are limited by economic conditions and cannot afford high-cost wastewater treatment equipment. Therefore, developing a multi-strain microbial-ceramsite carrier composite material that is highly adaptable, low-cost, and suitable for static nitrogen removal and purification is of great significance for achieving efficient nitrogen removal and purification of aquaculture wastewater in mountainous areas. Summary of the Invention

[0005] The purpose of this invention is to provide a method for preparing a multi-strain ceramsite carrier composite for static denitrification and purification of aquaculture wastewater and its application, thereby solving the problem of aquaculture wastewater pollution.

[0006] To achieve the above objectives, the present invention provides a multi-strain ceramsite carrier composite for static denitrification and purification of aquaculture wastewater, wherein, by mass percentage, the multi-strain ceramsite carrier composite comprises

[0007] Rhodopseudomonas palustris 20%–30%

[0008] Pediococcus pentaaceus 15%–30%

[0009] Bacillus licheniformis 10%–25%,

[0010] Candida albicans 10%–20%,

[0011] Leuconostoc mesenteroides 10%–15%

[0012] Cellulomonas flavigena 2%–5%

[0013] Staphylococcus saprophyticus 3%–5%;

[0014] The multi-strain-ceramsite carrier complex has a total bacterial count ≥1×10⁻⁶. 9 ~1×10 11 CFU / g.

[0015] To further explain, the multi-bacterial strain-ceramsite carrier composite material used for static denitrification and purification of aquaculture wastewater...

[0016] The preservation number of the aforementioned Rhodopseudomonas palustris is BMZ121828;

[0017] The preservation number of the *Pediococcus pentasaceus* is BMZ123303.

[0018] The accession number for the Bacillus licheniformis is BMZ137481.

[0019] The preservation number of the *Candida albicans* strain is BMZ134172.

[0020] The Leuconostoc mesenteroides species in question has the accession number CGMCC1.2138.

[0021] The preservation number of the *Cellulomonas flavigena* is BMZ134560.

[0022] The accession number for the saprophytic Staphylococcus saprophyticus is CICC24370.

[0023] This invention also provides a method for preparing the multi-species ceramsite carrier complex for static denitrification and purification of aquaculture wastewater, wherein *Rhodopseudomonas palustris* is cultured separately in a photosynthetic bacteria medium via liquid fermentation, *Pediococcus pentaaceus* is cultured separately in a high-density fermentation medium via liquid fermentation, *Bacillus licheniformis* is cultured separately in an LB+starch medium via liquid fermentation, and *Candida albicans* is cultured separately in a YPD medium via liquid fermentation. Liquid fermentation was performed. Leuconostoc mesenteroides was cultured alone on MRS modified medium, Cellulomonas flavigena was cultured alone on CMC medium, and Staphylococcus saprophyticus was cultured alone on nutrient broth medium. The number of viable bacteria was detected. After centrifugation, the fermentation broth was removed, and the bacteria were mixed with carrier materials to prepare a multi-strain-ceramsite carrier complex for static denitrification and purification of aquaculture wastewater.

[0024] To further explain, Candida albicans is part of the normal human flora and is usually found in the mouth, intestines, skin and vagina. It does not cause disease when the body is in normal condition, but when the body's immunity declines, Candida albicans may multiply in large numbers.

[0025] To further clarify, the *Staphylococcus saprophyticus* strain in question is a non-pathogenic strain that has undergone rigorous screening.

[0026] To further clarify, the seven strains underwent activation treatment before preparing the strain-carrier complex. *Rhodopseudomonas palustris* was activated using photosynthetic bacteria medium, *Pediococcus pentaaceus* using high-density fermentation medium, *Bacillus licheniformis* using LB+starch medium, *Candida albicans* using YPD medium, *Leuconostoc mesenteroides* using MRS modified medium, *Cellulomonas flavigena* using CMC medium, and *Staphylococcus saprophyticus* using nutrient broth medium. The activated bacterial solutions were then sequentially inoculated into 10%, 20%, 50%, and 100% sterilized aquaculture wastewater for acclimatization and culture. Finally, each strain was inoculated into its corresponding culture medium for cultivation and preserved as seed strains.

[0027] To further clarify, the sterilized aquaculture wastewater is obtained through a manure soaking and cleaning process;

[0028] The wastewater from pig farming includes, but is not limited to, wastewater from pig farms using the manure-soaking and cleaning process. This wastewater contains urine, feces, and pen flushing water, and contains no disinfectants, sodium hydroxide, or other drugs that are detrimental to the growth of microorganisms. It is also wastewater from pig farming discharged within the past week.

[0029] To further explain, the carrier material is ceramsite; the ceramsite is floating ceramsite and / or suspended ceramsite and / or sinking ceramsite; the carrier material can increase air permeability and compatibility and easily form a biofilm on its surface, which is conducive to bacterial attachment and metabolic activities.

[0030] Further explanation: The *Rhodopseudomonas swampensis* and the submerged ceramsite carrier are mixed at a bacterial culture volume: carrier mass ratio of 1:0.2 (L / kg) to prepare a strain-carrier complex; *Pediococcus pentosaceus* and the submerged ceramsite carrier are mixed at a bacterial culture volume: carrier mass ratio of 1:0.2 (L / kg) to prepare a strain-carrier complex; *Bacillus licheniformis* and the floating ceramsite carrier are mixed at a bacterial culture volume: carrier mass ratio of 1:0.2 (L / kg) to prepare a strain-carrier complex; *Candida albicans* and the suspended ceramsite carrier are mixed at a bacterial culture volume: carrier mass ratio of 1:0.4 (L / kg) to prepare a strain-carrier complex; *Leuconostoc mesenteroides* and the suspended ceramsite carrier are mixed at a bacterial culture volume: carrier mass ratio of 1:0.4 (L / kg) to prepare a strain-carrier complex; A bacterial strain-carrier complex was prepared by mixing 0.2 (L / kg) of the bacterial strain and carrier. The chlorogenic fibromona and the submerged ceramsite carrier were mixed at a ratio of bacterial volume to carrier mass of 1:1 (L / kg) to prepare the bacterial strain-carrier complex. The saprophytic staphylococcus and the suspended ceramsite carrier were mixed at a ratio of bacterial volume to carrier mass of 1:0.3 (L / kg) to prepare the bacterial strain-carrier complex.

[0031] This invention also provides a method for static denitrification and purification of aquaculture wastewater. The method involves adding the multi-bacterial strain-ceramsite carrier complex into a wastewater tank at a depth of 2 meters, but not limited to this depth. The multi-bacterial strain-ceramsite carrier complex is added in stages and at different times. The staged addition involves adding the complex sequentially to the bottom, middle, and top layers. Initially (T=0), the submerged microbial strain-carrier complex is added to the bottom layer of the water in the low dissolved oxygen / anaerobic zone (1.5-2 meters). At T=24 hours, the complex is added to the water in the medium-low dissolved oxygen zone (0.5-1.5 meters). Suspended bacterial culture-carrier complex is added to the middle layer; after T=36 hours, floating bacterial culture-carrier complex is added to the surface layer of the water in the 0-0.5 meter high dissolved oxygen zone; Bacillus licheniformis is added to the 0-0.5 meter surface high dissolved oxygen zone; Candida albicans, Leuconostoc mesenteroides, and Staphylococcus saprophyticus are added to the 0.5-1.5 meter middle layer low dissolved oxygen zone; Rhodopseudomonas palustris, Pediococcus pentosaceus, and Cellulosum xanthophylloma are added to the 1.5-2 meter bottom low dissolved oxygen / anaerobic zone to ensure that the bacterial strains in each layer remain in the suitable dissolved oxygen zone.

[0032] To further explain, the strains are activated before the multi-strain-ceramsite carrier complex is added to the wastewater; the activation solution includes activation solution I and activation solution II.

[0033] Activation solution I is the activation solution used for the strain Bacillus licheniformis, which consists of sterile water, 0.3% glucose, and 0.3% peptone. The strain-carrier complex of this strain is mixed with activation solution I at a mass-volume ratio of 1:10.

[0034] Activation solution II is the activation solution used for strains Rhodopseudomonas palustris, Pediococcus pentosaceus, Candida albicans, Leuconostoc mesenteroides, Cellulosum xanthogenes, and Staphylococcus saprophyticus. Activation solution II is composed of sterile water, 0.3% glucose, 0.5% sodium acetate, 0.2% ammonium chloride, 0.05% potassium dihydrogen phosphate, and 0.15% yeast extract. The strain-carrier complex of this strain is mixed with activation solution II at a mass-volume ratio of 1:10.

[0035] After mixing the bacterial strain-carrier complex with the activation solution, pour it into a transparent plastic container, ensuring that the carrier is completely submerged. The total proportion of the carrier should not exceed 1 / 3 of the volume of the mixture. Activate for 8-10 hours.

[0036] The dosage of the multi-strain-ceramsite carrier complex is 150-200g per cubic meter of wastewater, and it is added in stages and layers at different times.

[0037] To further explain, this multi-strain-ceramsite carrier composite material is suitable for static denitrification and purification of aquaculture wastewater. When used for static denitrification and purification of aquaculture wastewater, it is applied in a layered manner to match the environmental conditions at different depths of the water body. To ensure the activity of the strains and the denitrification effect, the temperature of the wastewater needs to be controlled within the range of 20℃-35℃, the pH value needs to be adjusted within the range of 6.5-8.5, and the C / N ratio of the wastewater needs to be adjusted to 5:2-8:1. The cover should be made of a light-transmitting material, or a 620-650 LED array should be arranged around the equipment with a light intensity ≥5000 lux and a daily illumination time of 8:00-18:00.

[0038] In summary, due to the adoption of the above technical solution, the beneficial effects of the present invention are:

[0039] (1) The combination of strains has full functional coverage and significant synergistic effect.

[0040] Surface layer (0–0.5 meters, high dissolved oxygen zone): Aerobic Bacillus licheniformis dominates the decomposition of organic nitrogen, providing ample substrate for the nitrification process.

[0041] Middle layer (0.5–1.5 meters, medium to low dissolved oxygen zone): Candida albicans decomposes complex organic matter such as starch and protein, and regulates dissolved oxygen to stabilize the middle layer environment; Staphylococcus saprophyticus efficiently degrades oils and nitrogen-containing organic matter; Leuconostoc mesenteroides consumes excess oxygen, preventing it from penetrating to the bottom layer, while enhancing flocculation and improving sludge settling performance.

[0042] Bottom layer (1.5–2 meters, low dissolved oxygen / anaerobic zone): Rhodopseudomonas palustris serves as the denitrification core, efficiently converting ammonia nitrogen and nitrite under low light conditions and degrading hydrogen sulfide, reducing odor production; Pediococcus pentosaceus inhibits pathogens by producing lactic acid and forms flocs to adsorb suspended matter; Xanthomonas xanthomonas decomposes cellulose to release carbon sources, providing energy support for denitrification.

[0043] Through the innovative combination and synergistic effect of the above-mentioned bacterial strains, this multi-strain-ceramsite carrier complex exhibits excellent denitrification and purification efficiency in static wastewater treatment: total nitrogen (TN) degradation rate of 86.2%, ammonia nitrogen (NH4⁺-N) degradation rate of 88.2%, and COD degradation rate of 79.2%.

[0044] (2) Microbial strain-carrier composite technology to achieve long-term stable purification. The microbial strain-ceramic aggregate carrier composite technology utilizes the porous structure of ceramic aggregate to provide a stable "habitat" for microorganisms, effectively solving the problem of easy loss and difficult colonization of microorganisms in sewage; at the same time, it realizes the slow release of microbial agents, significantly prolongs the effective action time, and improves the sustainability of purification effect.

[0045] (3) Adapt to aerobic environments and precisely activate strain functions

[0046] By adding microbial strains in layers, they are distributed to suitable water depths according to their oxygen requirements. This strategy effectively avoids oxygen competition among microbial communities and, by creating differentiated dissolved oxygen and light environments (such as providing weak light for Rhodopseudomonas marsh), enables various microbial communities to work together efficiently in suitable "workstations," thereby improving overall denitrification efficiency.

[0047] (4) Low operating costs and environmental risks

[0048] No complex aeration and mixing equipment is required, and maintenance is simple after deployment. The ceramsite carrier is reusable, effectively reducing the technical threshold and operating costs of wastewater treatment. At the same time, the bacterial strains are fixed in the ceramsite and will not flow out with the effluent, avoiding the ecological risks caused by the spread of foreign bacteria, making it more suitable for the actual conditions of rural farms in mountainous areas.

[0049] (5) Practice green governance and promote the "dual carbon" goal

[0050] Compared to energy-intensive physical denitrification (such as membrane separation) and chemical denitrification technologies, biological denitrification processes consume almost no energy, and the ceramsite carrier can be repeatedly regenerated (loaded with new microbial agents for recycling), reducing solid waste generation and conforming to the environmental protection concept of "low carbon and circularity".

[0051] Reducing pollutant emissions from livestock farming at the source will help control agricultural non-point source pollution (livestock farming pollution is one of the important sources of agricultural non-point source pollution), promote the transformation of agricultural production methods from "high pollution and high consumption" to "green and ecological", and provide practical support for the country's "dual carbon" goals and ecological civilization construction. Attached Figure Description

[0052] Figure 1 Examples 8, 9, and 10 illustrate the effects of multi-bacterial strain-ceramsite carrier composites on total nitrogen (TN) and ammonia nitrogen (NH4+) in the treatment of aquaculture wastewater. + Comparison of degradation rates of N-N and COD. Detailed Implementation

[0053] All features disclosed in this specification, or all steps in all disclosed methods or processes, may be combined in any way, except for mutually exclusive features and / or steps.

[0054] Unless otherwise stated, each feature disclosed in this specification (including any appended claims and abstract) is merely one example of a series of equivalent or similar features.

[0055] The strains used in this invention are from the following sources:

[0056] The preservation number of the aforementioned Rhodopseudomonas palustris is BMZ121828;

[0057] The preservation number of the *Pediococcus pentasaceus* is BMZ123303.

[0058] The accession number for the Bacillus licheniformis is BMZ137481.

[0059] The preservation number of the *Candida albicans* strain is BMZ134172.

[0060] The Leuconostoc mesenteroides species in question has the accession number CGMCC1.2138.

[0061] The preservation number of the *Cellulomonas flavigena* is BMZ134560.

[0062] The accession number for the saprophytic Staphylococcus saprophyticus is CICC24370.

[0063] All of the strains listed above are for sale and can be purchased on the market.

[0064] The culture medium composition formula used in this invention is as follows:

[0065] Oil culture medium: 1g peptone, 0.3g beef extract, 1g peanut oil, 0.5g sodium chloride, 0.01ml neutral red (1.6% aqueous solution), 2.8g agar powder, 100ml ultrapure water, pH 7.2-7.4, autoclaved at 121℃ for 20min.

[0066] Broth culture medium: 0.3% beef extract, 1.0% peptone, 0.5% NaCl, 100ml ultrapure water, pH at rest, autoclaved at 121℃ for 20min.

[0067] LB medium: 1g tryptone, 0.5g yeast extract, 0.5g sodium chloride, 100ml ultrapure water, pH at rest, autoclave at 121℃ for 20min.

[0068] Starch culture medium: 10g peptone, 3g beef extract, 5g sodium chloride, 28g agar powder, 2g soluble starch, 1000ml ultrapure water, pH 7.2-7.4, autoclaved at 121℃ for 20min.

[0069] Milk culture medium: 1000ml of nutrient agar (3% agar powder), autoclaved at 121℃ for 20min. 10g milk powder added to 200ml of ultrapure water, autoclaved at 121℃ for 20min. Finally, mix the nutrient agar and milk powder solution at a ratio of 100:20.

[0070] Photosynthetic bacteria culture medium: sodium acetate 3g, NH4Cl 1g, KH2PO4 0.5g, MgCl2 0.2g, yeast extract 0.1g / L.

[0071] High-density fermentation medium: 40g / L sugarcane molasses + 20g / L corn steep liquor powder + 5g / L sodium acetate.

[0072] LB+ Starch Medium: 10g peptone + 5g yeast extract + 5g soluble starch.

[0073] YPD medium: 3.0g yeast extract + 3.0g malt extract + 10.0g glucose + 5.0g casein peptone.

[0074] MRS medium: 10g casein peptone + 10g beef extract + 5g glucose + 5g sodium acetate + 1ml Tween 80.

[0075] CMC medium: 1.0% straw powder + 0.6% compound nitrogen source + 0.1% KH2PO4 + 0.1% MgSO4.

[0076] The following seven strains underwent activation treatment before preparing the strain-carrier complex. Rhodopseudomonas palustris was activated using photosynthetic bacteria medium, Pediococcus pentasaceus using high-density fermentation medium, Bacillus licheniformis using LB + starch medium, Candida albicans using YPD medium, Leuconostoc mesenteroides using MRS modified medium, Cellulomonas flavigena using CMC medium, and Staphylococcus saprophyticus using nutrient broth medium. The activated bacterial solutions were then sequentially inoculated into 10%, 20%, 50%, and 100% sterilized aquaculture wastewater for acclimatization and culture. Finally, each strain was inoculated into its corresponding culture medium for cultivation and preserved as seed strains.

[0077] The sterilized livestock wastewater is obtained through a manure soaking and cleaning process; the livestock wastewater includes, but is not limited to, wastewater from pig farms using the manure soaking and cleaning process, which includes urine, feces, and pen flushing water, and contains no disinfectants, sodium hydroxide, or other drugs that are detrimental to the growth of microorganisms, and is pig farm wastewater discharged within the past week.

[0078] Example 1: Preparation of a multi-strain microbial-ceramsite carrier composite for denitrification and purification of aquaculture wastewater

[0079] The specific method for preparing a multi-strain bacteria-cement granule carrier composite for denitrification and purification of aquaculture wastewater in this embodiment is as follows:

[0080] Rhodopseudomonas palustris was cultured separately in photosynthetic bacteria medium for liquid fermentation at 30°C under 5000 lux light for 72 hours. The OD at 600 nm was measured to be 3.5-4.5. After centrifugation, the fermentation broth was removed and diluted to a concentration ≥1×10⁻⁶. 9 CFU / mL bacterial suspension was mixed with the submersible ceramsite carrier at a ratio of bacterial suspension volume to carrier mass = 1:0.2 (L / kg) to prepare a bacterial strain-carrier complex, which is used to treat aquaculture wastewater for denitrification and purification.

[0081] Example 2: Preparation of a multi-strain microbial-ceramsite carrier composite for denitrification and purification of aquaculture wastewater

[0082] The specific method for preparing a multi-strain bacteria-cement granule carrier composite for denitrification and purification of aquaculture wastewater in this embodiment is as follows:

[0083] Pediococcus pentasaceus was cultured separately in high-density fermentation medium at 37°C using liquid fermentation. The OD600nm value was measured to be 2.0-3.0. After centrifugation, the fermentation broth was removed and diluted to a concentration ≥1×10⁻⁶. 9 A CFU / mL bacterial suspension was mixed with the aforementioned submersible ceramsite carrier at a ratio of bacterial suspension volume to carrier mass of 1:0.2 (L / kg) to prepare a bacterial strain-carrier complex; the resulting multi-strain ceramsite carrier complex was used for denitrification and purification of aquaculture wastewater.

[0084] Example 3: Preparation of a multi-strain microbial-ceramsite carrier composite for denitrification and purification of aquaculture wastewater

[0085] The specific method for preparing a multi-strain bacteria-cement granule carrier composite for denitrification and purification of aquaculture wastewater in this embodiment is as follows:

[0086] Bacillus licheniformis was cultured in LB medium with starch at 37°C and 180 rpm for 24 hours. The OD at 600 nm was measured to be 2.0-2.5. After centrifugation, the fermentation broth was removed and diluted to a concentration ≥1×10⁻⁶. 9 A CFU / mL bacterial suspension was mixed with the floating ceramsite carrier at a ratio of bacterial suspension volume to carrier mass of 1:0.2 (L / kg) to prepare a bacterial strain-carrier complex; the resulting multi-strain ceramsite carrier complex was used for denitrification and purification of aquaculture wastewater.

[0087] Example 4: Preparation of a multi-strain microbial-ceramsite carrier composite for denitrification and purification of aquaculture wastewater

[0088] The specific method for preparing a multi-strain bacteria-cement granule carrier composite for denitrification and purification of aquaculture wastewater in this embodiment is as follows:

[0089] Candida albicans was cultured statically in YPD medium at 28°C for 48 hours, followed by liquid fermentation. The OD at 600 nm was measured to be 3.0-4.0. After centrifugation, the fermentation broth was removed and diluted to a concentration ≥1×10⁻⁶. 9 CFU / mL bacterial suspension was mixed with the suspended ceramsite carrier at a ratio of bacterial volume to carrier mass = 1:0.4 (L / kg) to prepare a bacterial strain-carrier complex; thus, a multi-strain ceramsite carrier complex for treating aquaculture wastewater for denitrification and purification was obtained.

[0090] Example 5: Preparation of a multi-strain microbial-ceramsite carrier composite for denitrification and purification of aquaculture wastewater

[0091] The specific method for preparing a multi-strain bacteria-cement granule carrier composite for denitrification and purification of aquaculture wastewater in this embodiment is as follows:

[0092] Leuconostoc mesenteroides was cultured anaerobically at 30°C on MRS medium for 36 hours. After centrifugation, the fermentation broth was removed and diluted to a concentration ≥1×10⁻⁶. The OD₆₀nm value was measured at 1.5-2.0. 9 CFU / mL bacterial suspension was mixed with the suspended ceramsite carrier at a ratio of bacterial volume to carrier mass = 1:0.2 (L / kg) to prepare a bacterial strain-carrier complex, which is used to treat aquaculture wastewater for denitrification and purification.

[0093] Example 6: Preparation of a multi-strain microbial-ceramsite carrier composite for denitrification and purification of aquaculture wastewater

[0094] The specific method for preparing a multi-strain bacteria-cement granule carrier composite for denitrification and purification of aquaculture wastewater in this embodiment is as follows:

[0095] Cellulomonas flavigena was cultured in liquid form on CMC medium at 30°C, pH 7.0, 120 rpm for 48 h. The OD600 nm was measured at 1.8-2.2. After centrifugation, the fermentation broth was removed and diluted to a concentration ≥1×10⁻⁶. 9 CFU / mL bacterial suspension was mixed with the aforementioned submersible ceramsite carrier at a bacterial suspension volume: carrier mass = 1:1 (L / kg) to prepare a bacterial strain-carrier complex, which is then used to treat aquaculture wastewater for denitrification and purification.

[0096] Example 7: Preparation of a multi-strain microbial-ceramsite carrier composite for denitrification and purification of aquaculture wastewater

[0097] The specific method for preparing a multi-strain bacteria-cement granule carrier composite for denitrification and purification of aquaculture wastewater in this embodiment is as follows:

[0098] Staphylococcus saprophyticus was cultured statically in nutrient broth at 37°C for 24 hours, followed by liquid fermentation. The OD value at 600 nm was measured to be 1.5-1.8. After centrifugation, the fermentation broth was removed and diluted to a concentration ≥1×10⁻⁶. 9 CFU / mL bacterial suspension was mixed with the floating ceramsite carrier at a ratio of bacterial suspension volume to carrier mass = 1:0.3 (L / kg) to prepare a bacterial strain-carrier complex, which is used for denitrification and purification of aquaculture wastewater.

[0099] Example 8: Preparation of a multi-strain microbial-ceramsite carrier composite for denitrification and purification of aquaculture wastewater

[0100] The specific method for preparing a multi-strain bacteria-cement granule carrier composite for denitrification and purification of aquaculture wastewater in this embodiment is as follows:

[0101] Rhodopseudomonas palustris was cultured in liquid form only on photosynthetic bacteria medium; Pediococcus pentasaceus was cultured in liquid form only on high-density fermentation medium; Bacillus licheniformis was cultured in liquid form only on LB + starch medium; Candida albicans was cultured in liquid form only on YPD medium; Leuconostoc mesenteroides was cultured in liquid form only on MRS modified medium; Cellulomonas flavigena was cultured in liquid form only on CMC medium; and Staphylococcus saprophyticus was cultured in liquid form only on nutrient broth medium. The fermentation conditions were the same as in the previous examples. For steps 1, 2, 3, 4, 5, 6, and 7, the OD600nm values ​​were measured to be 3.5-4.5, 2.0-3.0, 2.0-2.5, 3.0-4.0, 1.5-2.0, 1.8-2.2, and 1.5-1.8, respectively. Then, a bacterial inoculum was prepared by mixing 20% ​​*Rhodopseudomonas palustris*, 15% *Pediococcus pentosaceus*, 25% *Bacillus licheniformis*, 20% *Candida albicans*, 15% *Leuconostoc mesenteroides*, 2% *Cellulomonas flavigena*, and 3% *Staphylococcus saprophyticus* according to their respective mass percentages. After centrifugation, the fermentation broth was removed, and the mixture was diluted to a concentration ≥1×10⁻⁶. 9 CFU / mL bacterial suspension was mixed with carrier material (7 strains were prepared according to the volume ratio of ceramsite carrier and bacterial suspension to carrier mass as in Examples 1-7), solidified and dried to obtain a multi-strain ceramsite carrier complex for denitrification and purification of aquaculture wastewater; total viable count ≥1×10⁻⁶. 9 -1×10 11 CFU / g.

[0102] Example 9: A Multi-Bacterial Species-Ceramic Particle Carrier Complex for Denitrification and Purification of Aquaculture Wastewater

[0103] The specific method for denitrification and purification of aquaculture wastewater using a multi-strain microbial-ceramsite carrier composite material in this embodiment is as follows:

[0104] Rhodopseudomonas palustris was cultured in liquid form only on photosynthetic bacteria medium; Pediococcus pentasaceus was cultured in liquid form only on high-density fermentation medium; Bacillus licheniformis was cultured in liquid form only on LB + starch medium; Candida albicans was cultured in liquid form only on YPD medium; Leuconostoc mesenteroides was cultured in liquid form only on MRS modified medium; Cellulomonas flavigena was cultured in liquid form only on CMC medium; and Staphylococcus saprophyticus was cultured in liquid form only on nutrient broth medium. The fermentation conditions were the same as in the previous examples. For steps 1, 2, 3, 4, 5, 6, and 7, the OD600nm values ​​were measured to be 3.5-4.5, 2.0-3.0, 2.0-2.5, 3.0-4.0, 1.5-2.0, 1.8-2.2, and 1.5-1.8, respectively. Then, a bacterial inoculum was prepared by mixing 25% *Rhodopseudomonas palustris*, 22% *Pediococcus pentaaceus*, 20% *Bacillus licheniformis*, 15% *Candida albicans*, 10% *Leuconostoc mesenteroides*, 5% *Cellulomonas flavigena*, and 3% *Staphylococcus saprophyticus* according to their respective mass percentages. After centrifugation, the fermentation broth was removed, and the mixture was diluted to a concentration ≥1×10⁻⁶. 9 CFU / mL bacterial suspension was mixed with carrier material (7 strains were prepared according to the volume ratio of ceramsite carrier and bacterial suspension to carrier mass as in Examples 1-7), solidified and dried to obtain a multi-strain ceramsite carrier complex for denitrification and purification of aquaculture wastewater; total viable count ≥1×10⁻⁶. 9 -1×10 11 CFU / g.

[0105] Example 10: A Multi-Bacterial Species-Ceramic Particle Carrier Composite for Denitrification and Purification of Aquaculture Wastewater

[0106] The specific method for preparing a multi-strain bacteria-cement granule carrier composite for denitrification and purification of aquaculture wastewater in this embodiment is as follows:

[0107] Rhodopseudomonas palustris was cultured in liquid form only on photosynthetic bacteria medium; Pediococcus pentasaceus was cultured in liquid form only on high-density fermentation medium; Bacillus licheniformis was cultured in liquid form only on LB + starch medium; Candida albicans was cultured in liquid form only on YPD medium; Leuconostoc mesenteroides was cultured in liquid form only on MRS modified medium; Cellulomonas flavigena was cultured in liquid form only on CMC medium; and Staphylococcus saprophyticus was cultured in liquid form only on nutrient broth medium. The fermentation conditions were the same as in the previous examples. For steps 1, 2, 3, 4, 5, 6, and 7, the OD600nm values ​​were measured to be 3.5-4.5, 2.0-3.0, 2.0-2.5, 3.0-4.0, 1.5-2.0, 1.8-2.2, and 1.5-1.8, respectively. Then, a bacterial inoculum was prepared by mixing 30% *Rhodopseudomonas palustris*, 30% *Pediococcus pentaaceus*, 10% *Bacillus licheniformis*, 10% *Candida albicans*, 10% *Leuconostoc mesenteroides*, 5% *Cellulomonas flavigena*, and 5% *Staphylococcus saprophyticus* according to their respective mass percentages. After centrifugation, the fermentation broth was removed, and the mixture was diluted to a concentration ≥1×10⁻⁶. 9 CFU / mL bacterial suspension was mixed with carrier material (7 strains were prepared according to the volume ratio of ceramsite carrier and bacterial suspension to carrier mass as in Examples 1-7), solidified and dried to obtain a multi-strain ceramsite carrier complex for denitrification and purification of aquaculture wastewater; total viable count ≥1×10⁻⁶. 9 -1×10 11 CFU / g.

[0108] Before the bacterial strain-carrier complexes prepared in Examples 1-10 are added to wastewater, the bacterial strains are activated; the activation solutions include Activation Solution I and Activation Solution II.

[0109] Activation solution I is the activation solution used for the strain Bacillus licheniformis, which consists of sterile water, 0.3% glucose, and 0.3% peptone. The strain-carrier complex of this strain is mixed with activation solution I at a ratio of 1:10 (mass-volume ratio).

[0110] Activation solution II is the activation solution used for strains Rhodopseudomonas palustris, Pediococcus pentosaceus, Candida albicans, Leuconostoc mesenteroides, Cellulosum xanthophyte, and Staphylococcus saprophyticus. Activation solution II is composed of sterile water, 0.3% glucose, 0.5% sodium acetate, 0.2% ammonium chloride, 0.05% potassium dihydrogen phosphate, and 0.15% yeast extract. The strain-carrier complex of these strains is mixed with activation solution II at a ratio of 1:10 (mass-volume ratio).

[0111] After mixing the microbial-carrier complex with the activation solution, pour it into a transparent plastic container, ensuring that the carrier is completely submerged. The total proportion of the carrier should not exceed 1 / 3 of the volume of the mixed solution. Activate for 8-10 hours. The dosage of the multi-microbial-ceramsite carrier complex is 150-200g per cubic meter of wastewater, and it should be added in stages and layers.

[0112] The bacterial strain-carrier complexes prepared in Examples 1-10 are all suitable for static denitrification and purification equipment for aquaculture wastewater. When this multi-strain-ceramsite carrier complex is used for static denitrification and purification of aquaculture wastewater, a layered delivery method is adopted to match the environmental conditions at different depths of the water body. To ensure the activity of the bacterial strains and the denitrification effect, the temperature of the wastewater needs to be controlled within the range of 20℃-35℃, the pH value needs to be adjusted within the range of 6.5-8.5, and the C / N ratio of the wastewater needs to be adjusted to 5:2-8:1. The cover should be made of a light-transmitting material or a 620-650 LED array should be arranged around the equipment with a light intensity ≥5000 lux and a daily illumination time of 8:00-18:00.

[0113] Experimental Example 1: Antagonism Detection

[0114] The seven types of fermentation and purification bacterial strains-ceramsite carrier complexes for wastewater treatment prepared in Examples 1-7 were each diluted to a concentration of 1×10⁻⁶. 8 CFU / mL bacterial suspension.

[0115] First, perforated Xinhua brand filter paper was prepared, and then packaged into several vaccine vials for sterilization; LB culture plates were prepared; the bacterial culture solution was diluted to 1×10⁻⁶. 8Add CFU / mL bacterial suspension to the sterilized vaccine vials, allowing the bacterial suspension to soak the filter paper. Use one vial of filter paper per bacterial suspension and label them. Take 0.1 mL of each bacterial suspension and spread it evenly on a plate. Spread each bacterial suspension on two plates. Using sterile forceps, place filter paper soaked in the suspension of another bacterial strain onto the plate. Place six filter paper pieces soaked in different bacterial suspensions on each plate and label them. Let stand for 30 minutes and incubate at 37℃ for 24 hours. Observe whether inhibition zones appear. If they appear, it indicates that there is an antagonistic effect between the two bacterial strains. The results are shown in Table 1.

[0116] Table 1 Results of antagonistic tests on different strains

[0117]

[0118] Note: " / " indicates no reaction between the same bacterial species; "-" indicates no inhibition zone between the two bacteria.

[0119] As shown in the table above, there was no antagonistic reaction between any two strains in the antagonism test.

[0120] Experimental Example 2: Detection of the Clear Zone on a Protein Plate

[0121] The seven strains prepared in Examples 1-7 for static denitrification and purification of aquaculture wastewater were dissolved in sterile water at a ratio of 1g:100mL. 10μL of each strain was inoculated onto milk agar plates using the streak plate method and incubated at 37℃ for 24h. The presence or absence of a clear zone around the colony was observed. A positive result was defined as a clear zone around the colony, while a negative result was defined as the absence of a clear zone. The positive strains were re-inoculated onto milk agar plates for 48h, and the ratio of the diameter of the hydrolyzed clear zone to the colony diameter was measured. The results are shown in Table 2.

[0122] As shown in Table 2, Examples 3, 4, and 6 exhibited a strong ability to break down proteins.

[0123] Experimental Example 3: Detection of the Transparent Zone on Starch Plates

[0124] The seven strains of multi-strain ceramsite carrier complexes for denitrification and purification of aquaculture wastewater prepared in Examples 1-7 were dissolved in sterile water at a ratio of 1g:100mL. 10μL of each strain was inoculated onto starch slurry plates using the streak plating method and incubated at 37℃ for 24h. Then, 0.8mL of 2% iodine tincture solution (pre-diluted 15 times with distilled water) was added to each plate. The plate was rotated to cover the entire surface with iodine solution, and allowed to stand for 1min. The presence of a colorless transparent zone around the colony was observed. A positive result was indicated by the appearance of a transparent zone, while a negative result was indicated by the absence of a transparent zone. Positive strains were re-inoculated onto starch slurry plates and incubated using the same procedure. The ratio of the diameter of the hydrolyzed transparent zone to the colony diameter was measured, and the results are shown in Table 2.

[0125] The results in Table 2 show that Examples 3, 4, and 6 exhibited a strong ability to decompose starch.

[0126] Experimental Example 4: Determination of Fat Hydrolysis Plate Detection and Oil Degradation Rate

[0127] The seven strains of bacteria prepared in Examples 1-7 for denitrification and purification of aquaculture wastewater multi-strain-ceramsite carrier complex were dissolved in sterile water at a ratio of 1g:100mL. 10μL of each strain was inoculated onto oil culture medium plates using the streak plate method and incubated at 37℃ for 24h. The results were observed. If the colonies were red and there were red spots where the colonies grew, it indicated that the fat had been hydrolyzed, and the result was considered positive. Otherwise, it was considered negative.

[0128] The strains that tested positive for the above-mentioned oil culture medium were picked up with an inoculation loop and transferred to Erlenmeyer flasks containing 100 ml of nutrient broth. The flasks were then shaken at 37 °C and 180 r / min for 24 h. The cultures were then inoculated again with a degradation medium composed of olive oil at a concentration of 10%, and fermented at 37 °C and 180 r / min for 2 days. The oil degradation rate was then measured.

[0129] The oil content was determined according to the method described by Qin Huaming (2003). The oil degradation rate was calculated according to the method described by Meng Zhuoni (2019). The results are shown in Table 2.

[0130] The results in Table 2 show that Examples 1, 3, 6 and 7 have a certain ability to degrade oils, but Example 7 shows a stronger ability to decompose oils.

[0131] Table 2. Results of tests on the degradation ability of different strains of oil, starch, and protein.

[0132]

[0133] Experiment 5: Denitrification and Purification Experiment of Aquaculture Wastewater

[0134] The wastewater came from a small-scale pig farm in Chongzuo City, which uses a process of cleaning manure by soaking it in water.

[0135] Before adding the multi-strain ceramsite carrier complex to the wastewater, the multi-strain ceramsite carrier complex is activated. The ceramsite carrier complex of each strain is mixed with the activation solution at a ratio of 1:10 (mass-volume ratio). The dosage of the multi-strain ceramsite carrier complex is 150-200g per cubic meter of wastewater, added in stages and layers. The wastewater temperature is controlled within the range of 20℃-35℃, the pH value is adjusted within the range of 6.5-8.5, and the C / N ratio is adjusted to 5:2-8:1. The multi-strain ceramsite carrier complex is added in stages and layers; this staged layering involves adding the complex sequentially to the bottom, middle, and surface layers. Initially (T=0), a submerged ceramsite carrier complex is added to the bottom layer (1.5-2 meters) of the low dissolved oxygen / anaerobic zone; at T=24 hours, a suspended ceramsite carrier complex is added to the middle layer (0.5-1.5 meters) of the medium-low dissolved oxygen zone; at T=36 hours, the remaining ceramsite carrier complex is added to the surface layer. Floating bacterial-carrier complexes are added to the surface layer of the water in the 0-0.5 meter high dissolved oxygen zone; Bacillus licheniformis is added to the 0-0.5 meter surface high dissolved oxygen zone; Candida albicans, Leuconostoc mesenteroides, and Staphylococcus saprophyticus are added to the 0.5-1.5 meter middle layer low dissolved oxygen zone; Rhodopseudomonas palustris, Pediococcus pentosaceus, and Cellulosum xanthophylloma are added to the 1.5-2 meter bottom low dissolved oxygen / anaerobic zone to ensure that the bacterial strains in each layer remain in the suitable dissolved oxygen zone.

[0136] The experiment included a blank control group (CK), in which only aquaculture wastewater was added;

[0137] Experiment 1 consisted of aquaculture wastewater plus the multi-strain ceramsite carrier complex from Example 8;

[0138] Experimental group 2 consisted of aquaculture wastewater plus the multi-strain ceramsite carrier complex from Example 9;

[0139] Experiment 3 consisted of aquaculture wastewater plus the multi-bacterial strain-ceramsite carrier complex from Example 10.

[0140] Each treatment group was repeated 3 times.

[0141] Each strain of bacteria was inoculated with its own strain-vector complex every 3 days for a total of 12 days. Sampling was performed before the start of the experiment and at 12 days. The main indicators detected were total nitrogen (TN), ammonia nitrogen (NH4⁺-N), and COD.

[0142] Total nitrogen (TN) content was determined according to HJ626-2012;

[0143] The determination of ammonia nitrogen (NH4⁺-N) content shall be in accordance with HJ525-2009;

[0144] The COD content was determined according to HJ828-2017, and the results are shown in [see attached table]. Figure 1 .

[0145] from Figure 1It can be seen that the multi-strain-ceramsite carrier complex in Example 9 has the best degradation effect. The total nitrogen (TN) degradation rate is 86.2%, while the blank control group is 26.1%, which is 3.3 times higher than the control group; the ammonia nitrogen (NH4⁺-N) degradation rate is 88.2%, while the blank control group is 28.3%, which is 3.1 times higher than the control group; and the COD degradation rate is 79.2%, while the blank control group is 22.2%, which is 3.57 times higher than the control group.

[0146] The above data indicate that the multi-strain-ceramsite carrier composite has a better effect on denitrification and purification of aquaculture wastewater.

[0147] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A multi-strain-ceramic carrier complex for static denitrification purification of aquaculture wastewater, characterized in that: The multi-strain-ceramic carrier complex comprises, in percentage by mass Rhodopseudomonas palustris 20% to 30%, Pediococcus pentasaceus 15% to 30%, Bacillus licheniformis 10% to 25%, Candida albicans 10% to 20%, Leuconostoc mesenteroides 10% to 15%, Cellulomonas flavigena 2% to 5%, Staphylococcus saprophyticus 3% to 5%. The multi-species-ceramic carrier complex, total bacteria amount ≥ 1 × 10 9 ~1 × 10 11 CFU / g.

2. The multi-strain-ceramic carrier complex for static denitrification of aquaculture wastewater according to claim 1, characterized in that: The accession number of the Rhodopseudomonas palustris is BMZ121828; The accession number of the Pediococcus pentasaceus is BMZ123303, The accession number of the Bacillus licheniformis is BMZ137481, The accession number of the Candida albicans is BMZ134172, The accession number of the Leuconostoc mesenteroides is CGMCC1.2138, The accession number of the Cellulomonas flavigena is BMZ134560, The accession number of the Staphylococcus saprophyticus is CICC24370.

3. A method for preparing a multi-strain-ceramic carrier complex for static denitrification purification of aquaculture wastewater as claimed in claim 1, characterized by: The Rhodopseudomonas palustris is cultured by liquid fermentation with a photosynthetic bacteria culture medium, the Pediococcus pentasaceus is cultured by liquid fermentation with a high-density fermentation culture medium, the Bacillus licheniformis is cultured by liquid fermentation with an LB+starch culture medium, the Candida albicans is cultured by liquid fermentation with a YPD culture medium, the Leuconostoc mesenteroides is cultured by liquid fermentation with a modified MRS culture medium, the Cellulomonas flavigena is cultured by liquid fermentation with a CMC culture medium, and the Staphylococcus saprophyticus is cultured by liquid fermentation with a nutrient broth culture medium, and then the effective viable cell count is detected, the fermentation liquid is removed after centrifugation, and the carrier material is mixed to prepare a bacteria-carrier composite, thereby obtaining a bacteria-carrier composite for breeding static denitrification purification of sewage.

4. The production method according to claim 3, characterized by: The seven strains are activated before being prepared into the bacteria-carrier composite, and the Rhodopseudomonas palustris is activated with the photosynthetic bacteria culture medium, the Pediococcus pentasaceus is activated with the high-density fermentation culture medium, the Bacillus licheniformis is activated with the LB+starch culture medium, the Candida albicans is activated with the YPD culture medium, the Leuconostoc mesenteroides is activated with the modified MRS culture medium, the Cellulomonas flavigena is activated with the CMC culture medium, and the Staphylococcus saprophyticus is activated with the nutrient broth culture medium, and then the activated bacterial liquid is inoculated into 10%, 20%, 50% and 100% of sterilized breeding sewage for domestication culture, and finally inoculated into the corresponding culture medium for culture, thereby serving as seed bacteria for preservation.

5. The production method according to claim 3, wherein The sterilized breeding sewage is obtained by the bubble manure cleaning process. The breeding sewage includes but is not limited to the bubble manure cleaning process used in pig farms, and the sewage includes urine, feces and barn flushing water, and does not contain disinfectants, sodium hydroxide and other drugs that are not conducive to the growth of bacterial agents, and is the sewage discharged within one week.

6. The production method according to claim 3, wherein The carrier material is ceramic particles, and the ceramic particles are floating ceramic particles, suspended ceramic particles and / or submerged ceramic particles, and the carrier material can increase the air permeability and compatibility and easily form a biological membrane on the surface, thereby being beneficial to the adhesion and metabolic activity of bacteria.

7. The production method according to claim 6, wherein The marshy red pseudomonas and the submerged ceramic carrier are mixed to prepare a bacteria-carrier compound with a bacteria liquid volume to carrier mass ratio of 1:0.2 (L / kg); the pentose pediococcus and the submerged ceramic carrier are mixed to prepare a bacteria-carrier compound with a bacteria liquid volume to carrier mass ratio of 1:0.2 (L / kg); the bacillus licheniformis and the floating ceramic carrier are mixed to prepare a bacteria-carrier compound with a bacteria liquid volume to carrier mass ratio of 1:0.2 (L / kg); the white Candida and the suspended ceramic carrier are mixed to prepare a bacteria-carrier compound with a bacteria liquid volume to carrier mass ratio of 1:0.4 (L / kg); the enteromorpha Leuconostoc and the suspended ceramic carrier are mixed to prepare a bacteria-carrier compound with a bacteria liquid volume to carrier mass ratio of 1:0.2 (L / kg); the xanthomonas filamentous bacteria and the submerged ceramic carrier are mixed to prepare a bacteria-carrier compound with a bacteria liquid volume to carrier mass ratio of 1:1 (L / kg); and the saprophytic staphylococcus and the suspended ceramic carrier are mixed to prepare a bacteria-carrier compound with a bacteria liquid volume to carrier mass ratio of 1:0.3 (L / kg).

8. A treatment method for static denitrification purification of aquaculture wastewater, characterized by, The multi-bacteria ceramic carrier compound prepared in claim 3 is put into, but not limited to, a 2-meter-deep sewage pool, and the multi-bacteria ceramic carrier compound is added in layers at different time periods; the layer-by-layer addition at different time periods is performed in layers from bottom to surface, at an initial time (T=0), the submerged bacteria-carrier compound is added to the water bottom layer in a low-oxygen / anaerobic zone at 1.5-2 meters; at T=24 hours, the suspended bacteria-carrier compound is added to the water middle layer in a medium-low-oxygen zone at 0.5-1.5 meters; at T=36 hours, the floating bacteria-carrier compound is added to the water surface layer in a high-oxygen zone at 0-0.5 meters; the bacillus licheniformis is added to the high-oxygen zone at 0-0.5 meters of the surface layer; the white Candida, the enteromorpha Leuconostoc, and the saprophytic staphylococcus are added to the medium-low-oxygen zone at 0.5-1.5 meters of the middle layer; the marshy red pseudomonas, the pentose pediococcus, and the xanthomonas filamentous bacteria are added to the low-oxygen / anaerobic zone at 1.5-2 meters of the bottom layer, so as to ensure that each layer of bacteria strain stays in a suitable oxygen zone.

9. The treatment method of claim 8, wherein, The multi-bacteria ceramic carrier compound is activated before being put into sewage; The activation liquid includes activation liquid I and activation liquid II; The activation liquid I is used for the bacillus licheniformis and is composed of sterile water, 0.3% glucose, and 0.3% protein peptone; the bacteria-carrier compound of the bacillus licheniformis is mixed with the activation liquid I at a mass-volume ratio of 1:10; The activation liquid II is used for the marshy red pseudomonas, the pentose pediococcus, the white Candida, the enteromorpha Leuconostoc, the xanthomonas filamentous bacteria, and the saprophytic staphylococcus; the activation liquid II is composed of sterile water, 0.3% glucose, 0.5% sodium acetate, 0.2% ammonium chloride, 0.05% potassium dihydrogen phosphate, and 0.15% yeast extract; the bacteria-carrier compound of the bacteria strain is mixed with the activation liquid II at a mass-volume ratio of 1:10; The bacteria-strain carrier compound is mixed with the activating liquid and poured into a transparent plastic container, ensuring that the carrier is completely immersed, and the total proportion of the carrier does not exceed 1 / 3 of the volume of the mixed liquid, and activated for 8-10 hours; The multi-strain-ceramic carrier compound is added in an amount of 150-200g per cubic meter of sewage, and is added in a time period and layer by layer.

10. The treatment method of claim 8, wherein: It is suitable for static nitrogen removal and purification equipment for aquaculture wastewater. When the multi-strain-ceramic carrier compound is used for static nitrogen removal and purification treatment of aquaculture wastewater, a layered adding method is adopted to match the environmental conditions at different depths of the water body. In order to ensure the activity of the strain and the nitrogen removal effect, the temperature of the wastewater needs to be controlled in the range of 20-35°C, the pH value needs to be controlled in the range of 6.5-8.5, and the C / N of the wastewater needs to be adjusted to 5:2-8:

1. The cover is a light-transmitting material or a 620-650 LED array is arranged around the equipment, the light intensity is ≥5000 lux, and the daily light time is 8:00-18:00.

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