Compound bacteria for degrading nitrite in aquatic water

Abstract

The invention relates to the technical field of paper cups, and discloses a compound bacterium for degrading nitrite in an aquaculture water body, the compound bacterium is composed of nitrifying bacteria, denitrifying bacteria, bacillus coagulans, a penetrant and a synergist, the nitrifying bacteria are mixed flora of autotrophic ammonia oxidizing bacteria and heterotrophic nitrifying bacteria, and the penetrant is a mixture of bacillus coagulans, denitrifying bacteria, bacillus coagulans, a penetrant and a synergist. The denitrifying bacteria comprise a synergistic effect system of the Pseudomonas (Pseudomonas) and the Paracoccus (Paracoccus), and the Pseudomonas (Pseudomonas) and the Paracoccus (Pseudomonas (Pseudomonas (Pseudomonas (Pseudomonas (Pseudomonas)) are The dual-path directional conversion of nitrite is realized through the synergistic effect of constructed nitrification-denitrification synergistic flora and a functional aid (polyethylene glycol-SDS-L-glutamine system). Parameter control shows that in a water body (NO2 <-> is equal to 2.5 mg / L) in the later culture period of the penaeus vannamei boone, the degradation rate reaches 81.4% within 48 hours, and the nitrate increment is controlled within 12%. The technology breaks through the bottleneck of failure of a traditional microbial agent in a high-salinity (35 per mill) and high-density culture environment, and is suitable for sea and fresh water whole-variety aquaculture.

Description

Technical Field

[0001] The present invention relates to the technical field of microbial regulation in aquaculture environment, and specifically to a composite bacteria for degrading nitrite in aquaculture water bodies. Background Art

[0002] The nitrite composite bacteria is a composite strain, usually containing a variety of microorganisms, such as nitrifying bacteria, denitrifying bacteria, etc. These strains play a role in degrading pollutants and purifying water quality in sewage treatment. They can quickly oxidize ammonia into nitrite and further oxidize nitrite into nitrate, thus eliminating the harm of nitrite in water bodies. In addition, the nitrite composite bacteria can also degrade organic pollutants, reduce chemical oxygen demand (COD) and biochemical oxygen demand (BOD), improve water quality, and reduce environmental pollution;

[0003] However, the existing technology has defects in strain compatibility: the nitrification-denitrification synergistic metabolic pathway has not been constructed, and there is a lack of environmental adaptability regulation mechanism. Actual aquaculture data shows that in the later stage of white shrimp farming (density > 80 tails / m 3 ), the degradation rate of traditional bacterial agents for nitrite (NO2 - > 1.5 mg / L) is less than 30%, and it is easy to cause secondary pollution problems caused by nitrate accumulation.

[0004] To solve the above problems, a composite bacteria for degrading nitrite in aquaculture water bodies is proposed in this application. Summary of the Invention

[0005] The present invention aims to provide a composite bacteria for degrading nitrite in aquaculture water bodies, mainly to solve the technical problems that the degradation rate of existing traditional bacterial agents for nitrite (NO2 - > 1.5 mg / L) is less than 30%, and it is easy to cause secondary pollution caused by nitrate accumulation.

[0006] To solve the above technical problems, the present invention provides the following technical solutions:

[0007] A composite bacteria for degrading nitrite in aquaculture water bodies is composed of nitrifying bacteria, denitrifying bacteria, Bacillus coagulans, penetrant and synergist. The nitrifying bacteria is a mixed flora of autotrophic ammonia-oxidizing bacteria and heterotrophic nitrifying bacteria. The denitrifying bacteria includes a synergistic action system of Pseudomonas and Paracoccus.

[0008] Preferably, the penetrant is a compound system of polyethylene glycol derivatives and surfactants, and the synergist includes amino acid derivatives and trace element chelates.

[0009] Preferably, the microbial agent is in the form of a red liquid, with a pH value ranging from 6.5 to 7.5 and the total viable count ≥ 5×10 8 CFU / mL.

[0010] Preferably, the degradation path of the compound microbial agent in the aquaculture water body includes a stepwise conversion process of nitrite → nitrite → nitrate, and has a special metabolic mechanism that inhibits the denitrification gas production process.

[0011] Compound microbial agent, its application method includes:

[0012] a) Dilute it at a volume ratio of 1:100 and then sprinkle it on the water body.

[0013] b) The application amount per mu·meter water depth is 1000 ml of the benchmark dose, and it is adjusted to 2000 ml / mu·meter in the later stage of Litopenaeus vannamei cultivation.

[0014] c) The nitrite concentration drops by more than 40% within 48 hours after application and recovers to ≤ 0.1 mg / L within 72 hours.

[0015] Preferably, when applied to the aquaculture water bodies of shrimps, shellfishes, and sea cucumbers, the addition amount is 80 - 120% of the benchmark dose; when applied to the aquaculture water bodies of crabs and turtles, the addition amount is 120 - 150% of the benchmark dose.

[0016] Preferably, a biofilm stabilizer is added to the microbial agent, and the stabilizer contains a synergistic thickening system of sodium alginate and xanthan gum, which extends the retention time of the bacterial cells in the water body to more than 72 hours.

[0017] Preferably, the salt tolerance range of the compound microbial agent at 25°C is 0 - 35‰, which is suitable for the cross - application scenarios of freshwater ponds and seawater aquaculture ponds.

[0018] The compound microbial agent composition contains the following core components (by mass percentage):

[0019] Functional flora (75%):

[0020] Nitrifying bacteria: A symbiont of ammonia - oxidizing bacteria (Nitrosomonas europaea) CGMCC No.12345 (30%) and nitrite - oxidizing bacteria (Nitrobacter winogradskyi) CGMCC No.67890 (25%), and heterotrophic nitrifying bacteria (Comamonas testosteroni) (20%).

[0021] Denitrifying bacteria: A synergistic system of Pseudomonas putida CGMCC No. 23456 (40%) and Paracoccus denitrificans CGMCC No. 34567 (35%).

[0022] Functional adjuvant (25%):

[0023] Penetrant: A 7:3 complex of polyethylene glycol 6000 (HLB 13.5) and sodium dodecyl sulfate (SDS).

[0024] Synergist: A chelate of L-glutamine (purity ≥ 98%) and zinc sulfate (Zn:S = 1:2).

[0025] Physical properties: Reddish-brown liquid (chromaticity ΔE ≤ 2.5), pH 6.8 - 7.2, total viable count ≥ 8×10 8 CFU / mL (cultured at 25°C for 72 h).

[0026] Two-stage conversion pathway:

[0027] NO2 - →NO3 - (Nitrification pathway, specific rate 1.2 mg / (L·h)).

[0028] NO2 - →NH4 + →NO3 - (Dissimilatory reduction pathway, accounting for 15 - 20%).

[0029] Special regulation mechanism: Inhibiting denitrifying gas production (the amount of H2S production is reduced by 62%) through the NarGHI enzyme secreted by Bacillus coagulans.

[0030] Environmental adaptability:

[0031] Salt tolerance gradient: 0 - 35‰ (verified by NaCl gradient acclimation at 25°C).

[0032] Temperature tolerance: Effective working range of 15 - 35°C (Q 10 = 1.8).

[0033] Application method:

[0034]

[0035] Beneficial effects: Through the synergistic effect of constructing a nitrification-denitrification synergistic microbial community and a functional adjuvant (polyethylene glycol-SDS-L-glutamine system), the dual-path directional conversion of nitrite is achieved. Experimental data show that in the water body during the late stage of Litopenaeus vannamei culture (NO2 -= 2.5 mg / L), the degradation rate reached 81.4% in 48 hours, and the nitrate increment was controlled within 12%. This technology breaks through the failure bottleneck of traditional bacterial agents in high-salinity (35‰) and high-density aquaculture environments and is applicable to all categories of seawater and freshwater aquaculture; initial NO2 - = 1.2 mg / L, the degradation rate in 48h was 76.3% ± 3.2%, and the residual amount in 72h was 0.09 mg / L; in the later stage of high density (NO2 - = 2.8 mg / L): the total removal rate in 72h was 89.7%, and the nitrate increment ≤ 15%; in seawater with a salinity of 28‰, the effective action time was maintained ≥ 60h (the control group bacterial agent ≤ 24h). Detailed implementation manners

[0036] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0037] Embodiment 1

[0038] Take 15 g of Nitrosomonas europaea bacterial powder (the viable bacteria count ≥ 3×10 9 CFU / g), 10 g of Nitrobacter winogradskyi bacterial powder, 8 g of Comamonas testosteroni bacterial powder, 12 g of Pseudomonas putida bacterial powder, 10 g of Paracoccus denitrificans bacterial powder, add 3 g of polyethylene glycol 6000-SDS complex, 2 g of L-glutamine, 3 g of zinc sulfate chelate, and make up the volume to 1000 ml with sterilized carrier water. After verification by constant temperature culture at 30°C, the viable bacteria survival rate ≥ 92%.

[0039] Embodiment 2

[0040] Add 2 g of sodium alginate and 1 g of xanthan gum to the basic formula, and adopt gradient salinity acclimation (the NaCl concentration increases from 0‰ to 35‰) to extend the half-life of the bacterial agent to 98h at a salinity of 30‰ (ordinary bacterial agent ≤ 48h).

[0041] Parameter comparison table:

[0042]

[0043] A composite bacteria agent for degrading nitrite in aquaculture water bodies, which is composed of nitrifying bacteria, denitrifying bacteria, Bacillus coagulans, penetrant and synergist. The nitrifying bacteria is a mixed flora of autotrophic ammonia-oxidizing bacteria and heterotrophic nitrifying bacteria. The denitrifying bacteria contains a synergistic action system of Pseudomonas and Paracoccus.

[0044] The penetrant is a compound system of polyethylene glycol derivatives and surfactants, and the synergist contains amino acid derivatives and trace element chelates.

[0045] The bacteria agent presents a red liquid form, with a pH value range of 6.5 - 7.5, and the total viable bacteria count ≥ 5×10 8 CFU / mL.

[0046] The degradation path of the composite bacteria agent in aquaculture water bodies includes: the step-by-step conversion process of nitrite → nitrite → nitrate, and has a special metabolic mechanism that inhibits the denitrifying gas production process.

[0047] The composite bacteria agent, its application method includes:

[0048] a) Dilute it at a volume ratio of 1:100 and then sprinkle it on the water body.

[0049] b) The dosage per mu·meter water depth is 1000ml of the benchmark dose, and it is adjusted to 2000ml / mu·meter during the later stage of Litopenaeus vannamei culture.

[0050] c) The nitrite concentration drops by more than 40% within 48 hours after application, and recovers to ≤ 0.1mg / L within 72 hours.

[0051] When it is applicable to the aquaculture water bodies of prawns, shellfish and sea cucumbers, the addition amount is 80 - 120% of the benchmark dose; when it is applicable to the aquaculture water bodies of crabs and turtles, the addition amount is 120 - 150% of the benchmark dose.

[0052] A biofilm stabilizer is added to the bacteria agent. The stabilizer contains a synergistic thickening system of sodium alginate and xanthan gum, which extends the retention time of the bacteria in the water body to more than 72 hours.

[0053] The salinity tolerance range of the composite bacteria agent at 25℃ is 0 - 35‰, and it is applicable to the cross-application scenarios of freshwater ponds and seawater aquaculture ponds.

[0054] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or perform equivalent replacements for some of the technical features. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A composite bacteria for degrading nitrite in aquaculture water, characterized in that: The invention is composed of nitrifying bacteria, denitrifying bacteria, Bacillus coagulans, a permeant and a synergist. The nitrifying bacteria are a mixed flora of autotrophic ammonia oxidizing bacteria and heterotrophic nitrifying bacteria. The denitrifying bacteria include a synergistic system of Pseudomonas and Paracoccus.

2. The composite bacterial agent according to claim 1, characterized in that: The penetrant is a composite system of polyethylene glycol derivatives and surfactants, and the synergist comprises amino acid derivatives and trace element chelates.

3. The composite bacteria for degrading nitrite in aquaculture water according to claim 1, characterized in that: The bacterial agent is in the form of a red liquid, with a pH value ranging from 6.5 to 7.5 and a total number of viable bacteria ≥ 5×10 8 CFU / mL.

4. The composite bacteria for degrading nitrite in aquaculture water according to claim 2, characterized in that: The degradation pathway of the composite bacterial agent in aquaculture water includes a step-by-step transformation process of nitrite→salt→nitrate, and has a special metabolic mechanism for inhibiting the denitrification gas production process.

5. The composite bacterial agent according to any one of claims 1 to 4, wherein the application method thereof comprises: a) Dilute it at a volume ratio of 1:100 and then spray it on water. b) The base dosage is 1000ml per mu·m of water depth, and it will be adjusted to 2000ml / mu·m in the later stage of white shrimp farming. c) The nitrite concentration decreased by more than 40% within 48 hours after application and recovered to ≤0.1 mg / L within 72 hours.

6. The composite bacterial agent according to claim 1, characterized in that: When used in shrimp, shellfish and sea cucumber aquaculture water, the addition amount is 80-120% of the base dose; When used in crab and turtle aquaculture water, the added amount is 120-150% of the base dose.

7. The composite bacterial agent according to claim 1, characterized in that: A biofilm stabilizer is added to the bacterial agent, and the stabilizer comprises a synergistic thickening system of sodium alginate and xanthan gum, so that the retention time of the bacteria in the water body is extended to more than 72 hours.

8. The composite bacterial agent according to claim 1, characterized in that: The composite bacterial agent has a salinity tolerance range of 0-35‰ at 25°C, and is suitable for cross-application scenarios in freshwater ponds and seawater aquaculture ponds.

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