Composite flora for improving water quality-aquatic product quality of ecological aquaculture and preparation and use methods thereof

Abstract

The invention discloses a composite flora for improving water quality of ecological aquaculture and aquatic product quality as well as a preparation method and a use method of the composite flora, and belongs to the technical field of microbial agents. The composite flora is composed of three modular flora bodies composed of multiple functional microbial strains, the three modular flora bodies are formed through a specific synergistic matching technology, the composite flora can be applied in a single water body splashing mode, and an innovative in-situ biological flocculation delivery synergistic technology is applied. According to the composite flora, the technical idea of'in-situ biological flocculation delivery 'is proposed and realized, that is, exopolysaccharide is secreted through the flora of the module I to construct a biological flocculation carrier, probiotics of the module III are embedded, and efficient colonization of intestinal tracts is realized through ingestion of a host. The synergistic water purification mechanism of substrate relay and the synergistic antibacterial mechanism of environmental modification and multi-target attack serve as the basis of efficient operation of the system.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of microbial agents, and specifically relates to a compound microbial community for improving water quality and aquatic product quality in ecological aquaculture and a preparation and use method thereof. BACKGROUND

[0002] Currently, high-density aquaculture modes generally face two interrelated technical challenges.

[0003] First, the continuous deterioration of the farming environment. A large amount of organic waste such as leftover feed and feces generated during the farming process leads to a serious over-standard concentration of harmful substances such as organic pollutants (measured by COD), ammonia nitrogen (NH4 + -N), and nitrite (NO2 ～ -N) in the water body. These pollutants not only directly poison the farmed animals, inhibit their growth, and even cause death, but also easily induce harmful algae outbreaks, further disrupting the ecological balance of the water body.

[0004] Second, the health and quality problems of the farmed products themselves. Pathogenic microorganisms are prone to breed in high-density environments, leading to frequent diseases. To control diseases, antibiotics are commonly used, but this brings a series of problems such as drug residues, enhanced pathogen resistance, and food safety risks of aquatic products. At the same time, the low immunity and poor digestive function of the farmed animals also directly affect their survival rate, growth rate, and final product quality.

[0005] In the prior art, two independent operations are usually needed to solve the above problems: ① spraying water purification bacteria such as nitrifying bacteria into the water body to regulate the water quality; and ② adding intestinal probiotics such as lactic acid bacteria through the method of mixed feeding to improve the intestinal health of animals. This "two-step method" mode has the following significant shortcomings:

[0006] (1) Functional fragmentation, limited application method: water purification products only act on the water body and cannot directly improve the health status of the animals. The application method (mixed feeding) of intestinal probiotics is cumbersome, and there are problems such as uneven feeding and loss in water. More importantly, if intestinal probiotics are directly sprayed into the water, the bacterial bodies will be diluted by the huge water body, and cannot effectively enter the animal intestine and form a dominant population, which is a fundamental technical problem, and basically belongs to invalid application. This makes the two functions fragmented in the technical path, and cannot form a systematic solution.

[0007] (2) Poor synergy, unstable effect: simple strain mixing products on the market, even if they contain water purification bacteria and probiotics, also lack internal synergy mechanisms, and their effect is greatly reduced and their performance is unstable in complex farming environments.

[0008] (3) The existing technology is a passive treatment, which cannot fundamentally build a "healthy environment promotes healthy aquaculture, and healthy aquaculture in turn benefits the healthy environment" virtuous ecological circulation system.

[0009] Therefore, there is an urgent need in the art for a revolutionary single product and single administration regimen that can break through the bottleneck of the prior art and efficiently and synergistically integrate water quality regulation and intestinal health improvement of aquatic animals. SUMMARY

[0010] The first object of the present application is to provide a composite microbial flora for improving the water quality and water product quality of ecological aquaculture, which is composed of three module microbial floras composed of multiple functional microbial strains, and is prepared by using the three module microbial floras in a specific synergistic ratio.

[0011] The second object of the present application is to provide a use method of a composite microbial liquid for improving the water quality and water product quality of ecological aquaculture, which is prepared by using module microbial floras, and can be directly applied to the aquaculture water.

[0012] The present application is achieved by the following technical solutions:

[0013] A composite microbial flora for improving the water quality and water product quality of ecological aquaculture, which is composed of three module microbial floras composed of multiple functional microbial strains, and is prepared by using the three module microbial floras in a specific synergistic ratio.

[0014] The environmental purification and bioflocculation carrier module microbial flora is composed of equal proportions of Bacillus subtilis and Bacillus licheniformis;

[0015] The pollutant deep conversion and water quality stabilization module microbial flora is composed of equal proportions of nitrifying bacteria and photosynthetic bacteria;

[0016] The host health directional gain and quality improvement module microbial flora is composed of equal proportions of Lactobacillus plantarum and Bacillus amyloliquefaciens.

[0017] The host health directional gain and quality improvement module microbial flora is composed of equal proportions of Lactobacillus plantarum and Bacillus amyloliquefaciens.

[0018] As a preferred, the composite microbial flora is composed of 50-70% of the environmental purification and bioflocculation carrier module microbial flora, 15-25% of the pollutant deep conversion and water quality stabilization module microbial flora, and 15-25% of the host health directional gain and quality improvement module microbial flora.

[0019] The environmental purification and bio-flocculation carrier module bacterial population is 60%, the pollutant deep conversion and water quality stabilization module bacterial population is 20%, and the host health directional gain and quality improvement module bacterial population is 20%.

[0020] As preferably, the nitrifying bacterial population is obtained by mixing equal proportions of Nitrosomonas and Nitrobacter;

[0021] The photosynthetic bacterial population is obtained by mixing equal proportions of Rhodopseudomonas, purple sulfur bacteria, green sulfur bacteria, cyanobacteria and halobacteria.

[0022] As preferably, the Bacillus amyloliquefaciens is Bacillus amyloliquefaciens FZB42.

[0023] A composite bacterial liquid for improving the water quality and water product quality of ecological aquaculture, the composite bacterial liquid comprising the composite bacterial population.

[0024] A preparation method of a composite bacterial liquid for improving the water quality and water product quality of ecological aquaculture, comprising the following steps:

[0025] S1, respectively, liquid fermentation culture of environmental purification and bio-flocculation carrier module bacterial population, pollutant deep conversion and water quality stabilization module bacterial population and host health directional gain and quality improvement module bacterial population, respectively, to obtain environmental purification and bio-flocculation carrier module bacterial population mother liquor, pollutant deep conversion and water quality stabilization module bacterial population mother liquor and host health directional gain and quality improvement module bacterial population mother liquor;

[0026] S2, according to the corresponding proportion, the environmental purification and bio-flocculation carrier module bacterial population mother liquor, the pollutant deep conversion and water quality stabilization module bacterial population mother liquor and the host health directional gain and quality improvement module bacterial population mother liquor are mixed to obtain the composite bacterial liquid.

[0027] As preferably, the viable bacterial concentration in the environmental purification and bio-flocculation carrier module bacterial population mother liquor, the pollutant deep conversion and water quality stabilization module bacterial population mother liquor, and the host health directional gain and quality improvement module bacterial population mother liquor is not less than 1×10 10 CFU / ml.

[0028] A use method of a composite bacterial liquid for improving the water quality and water product quality of ecological aquaculture, the composite bacterial liquid is embedded by hydrogel microcapsules, and is directly and uniformly sprinkled on the surface of the aquaculture water body.

[0029] As preferably, based on the first use or water quality deterioration of the aquaculture water body, 1L of composite bacterial liquid is used for 1000 cubic meters of water body.

[0030] As preferably, based on the daily maintenance of the aquaculture water body, 0.5L of composite bacterial liquid is used for 1000 cubic meters of water body every 10-15 days.

[0031] Compared with the prior art, the present application has at least the following technical effects:

[0032] The present application provides a composite microbial flora for improving water quality and aquatic product quality in ecological aquaculture, which is composed of three module microbial floras of multiple functional microbial strains, and is applied by a single water body spraying method and an innovative "in-situ biological flocculation delivery" synergistic technology.

[0033] The composite microbial flora has the following advantages:

[0034] (1) By scientific compounding of a high proportion (50% to 70%) of an environmental purification and biological flocculation carrier module flora (module one) and a professional pollutant deep conversion and water quality stabilization module flora (module two), an efficient external purification chain of "macromolecular organic matter degradation ~ directional conversion of toxic ammonia nitrogen" is constructed, and the problem of low efficiency of single water purification bacteria due to substrate limitation is solved.

[0035] Mechanism: The functional limitation of specific water purification bacterial strains such as nitrifying bacteria is mainly due to the specificity of their metabolic pathways: such bacterial flora can only metabolize small molecular ammonia nitrogen through the ammonia oxidation pathway, and lacks direct degradation ability for macromolecular organic pollutants such as leftover feed and feces commonly existing in water bodies.

[0036] The core innovation of the present technology is to systematically biodegrade and convert complex organic matter through efficient configuration of module one, so as to form ammonium salt products that can be directly utilized by module two flora. This phased synergistic metabolic mechanism constructs a complete pollutant conversion pathway, effectively solving the problem of microbial metabolic blockage caused by broken substrate conversion chain in traditional processes. By establishing a hierarchical progressive substrate delivery system, the metabolic coupling of different functional bacterial floras is realized, thereby breaking through the technical bottleneck of low efficiency of functional bacterial flora in existing biological water purification technology. This compounding method greatly improves the overall degradation rate of COD and ammonia nitrogen in water bodies. Combined with experimental data, the degradation rate of ammonia nitrogen of the present composite microbial flora is more than 30% higher than the arithmetic sum of the effects of module one and module two alone, showing a significant and unexpected synergistic effect, thereby more quickly and more thoroughly purifying the aquaculture water quality.

[0037] (2) Through the directional regulation of the Bacillus colony metabolic characteristics of module one, it can exert the biological polymerization function, and the host health directional gain and quality improvement module bacteria colony (module three) probiotics (plant lactic acid bacteria and Bacillus amyloliquefaciens FZB42 strain) and organic debris in the water body are co-polymerized to form a stable complex, and a composite system of "biological flocculation matrix ~ functional bacteria colony directional transmission" is constructed, which systematically breaks through the common problem that traditional water body spraying probiotics cannot be targeted to the host.

[0038] Mechanism: Unlike the theoretical assumption of traditional cognition that water free probiotics are quickly inactivated, this design is based on the characteristics of the extracellular polymeric substance (EPS) secreted by Bacillus during metabolism to realize functional coupling. This kind of high molecular substance can specifically adsorb suspended particles in water (including leftover debris, fecal particles and module three probiotics), and form a biological flocculation body with probiotics as the core and organic matter as the carrier through intermolecular forces. The composite structure is ecologically adapted to the filter-feeding habit of aquatic animals, and the functional bacteria colony is targeted delivered through the nutrition level.

[0039] This matching mode breaks through the technical formula that probiotics must be added through feed, and creates a water body probiotic biological efficiency delivery mode. By constructing an in-situ biological carrier, discrete bacterial agents are converted into functional nutritional units that are actively ingested by animals, achieving exponential improvement of intestinal colonization efficiency of bacteria colony. This technical path effectively solves the long-standing problem of functional attenuation in the field of water body probiotic application, and provides a breakthrough methodological paradigm for microbial regulation in aquaculture.

[0040] (3) Through the establishment of a self-sustaining cycle system of "ecological regulation-function metabolism coupling", the problem of low operating efficiency caused by the separation of traditional water quality management and health regulation modules is systematically solved. Based on the core mechanism of biological flocculation, the functional synergy of water pollutant reduction and probiotic delivery is innovatively realized through physical and chemical processes. Mechanism: The biological flocculation system has a dual regulation mechanism: first, through the charge neutralization and bridging aggregation effect of organic particles, the mineralization rate of heterotrophic bacteria on pollutants is significantly improved (COD degradation efficiency is increased by 20%, and ammonia nitrogen conversion period is shortened by 54%); second, the microecological carrier formed is adapted to the size of the feeding behavior of aquatic animals (50-200 μm), and the intestinal colonization concentration of the bacteria colony (Lactobacillus plantarum) reaches 2.8×10 7 CFU / g, which is three orders of magnitude (about 1800 times) higher than the traditional spraying control group (1.5×10 4 CFU / g), breaking through the effective colonization threshold of probiotics (10 6 CFU / g), and confirming the innovation of the technical path.

[0041] The use mode synchronously realizes efficient delivery of probiotics and comprehensive water quality regulation through a "one-time spraying" innovative mode. The scheme integrates the colonization of microbial flora, water quality improvement, and activation of nutrient metabolism into a single operation through a "carrier-behavior-metabolism" three-level synergistic mechanism, completely changing the traditional repeated spraying mode, and the operation efficiency is improved by more than 1000 times. The commercial value has been verified in large-scale aquaculture, and the water product quality is upgraded to intensive and standardized management.

[0042] The composite bacteria liquid prepared from the composite microbial flora for improving the water quality and water product quality of ecological aquaculture does not need to distinguish between water purifying agents and feed additives in use. Only by one-time water spraying, the water purification and animal intestinal health improvement programs can be started at the same time, greatly reducing the labor intensity and management cost. BRIEF DESCRIPTION OF DRAWINGS

[0043] Figure 1 The figure is a schematic diagram of Lactobacillus plantarum colony for grass carp aquaculture experiment and intestinal flora detection using the bacteria liquid prepared in Example 3.

[0044] Figure 2 The figure is a schematic diagram of a gram staining microscope photo of the colony plate. DETAILED DESCRIPTION

[0045] The embodiments of the present application will be described in detail below with reference to the examples, but those skilled in the art will understand that the following examples are only for illustration of the present application and should not be regarded as limiting the scope of the present application. The specific conditions not mentioned in the examples are carried out according to the conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments not mentioned by the manufacturer are all conventional products that can be purchased on the market.

[0046] The technical solution of one specific embodiment of the present application is:

[0047] The present application provides a synergistic composite microbial flora, characterized in that it is composed of microbial strains of the following three functional modules, and is compounded according to a specific viable count (Colony-Forming Units, CFU) ratio.

[0048] Module 1: Environmental purification and bioflocculation carrier construction module flora, accounting for 50% to 70% of the total viable count.

[0049] Composition: composed of Bacillus subtilis and Bacillus licheniformis mixed at a ratio of 1:1.

[0050] Action and mechanism:

[0051] (1) High-yield hydrolytic enzymes to degrade organic matter (reduce COD): Both of the two Bacillus can efficiently secrete proteases, amylases, lipases and cellulases. These enzymes can quickly hydrolyze large molecular organic matter such as suspended and sedimentary residues and feces in the water body into small molecular amino acids and monosaccharides, thereby directly and quickly reducing the chemical oxygen demand (COD) of the water body, improving the water color and reducing oxygen consumption.

[0052] (2) Provide substrate for nitrogen cycle: The amino acids produced by the above hydrolysis process will further be deaminated to generate ammonia nitrogen (NH3 / NH4 + ), providing a continuous substrate for the downstream nitrogen cycle module.

[0053] (3) Construction of bioflocculation carrier (core innovation): The two Bacillus will secrete adhesion substances such as extracellular polysaccharides (EPS) during growth and metabolism, which will coagulate suspended organic matter, microalgae and complex bacterial flora in the water body through physical-chemical synergistic effect, forming high-nutrient-density bioflocculation, and simultaneously achieving pollution enrichment and targeted delivery of probiotics.

[0054] Parameter optimization basis: The module ratio of 50% to 70% is the best balance interval for the system. If it is lower than 50%, the organic load degradation rate is insufficient, leading to substrate shortage in module two, and decreased purification efficiency; if it is higher than 70%, the ecological niche of modules two and three is squeezed, weakening the ammonia nitrogen deep conversion and host gain function.

[0055] Module two: Pollutant deep conversion and water quality stabilization module bacterial flora, accounting for 15% to 25% of the total viable bacterial count.

[0056] Composition: The nitrifying bacterial flora composed of Nitrosomonas and Nitrobacter and the photosynthetic bacterial flora such as Rhodopseudomonas are mixed in a ratio of 1:1.

[0057] Action and mechanism:

[0058] (1) Aerobic nitrification: Through ammonia oxidation (NH4 + → NO2 - ) and nitrite oxidation (NO2 - → NO3 - ), ammonia nitrogen is converted in stages, eliminating the acute toxicity of the aquaculture water body.

[0059] (2) Anaerobic / micro-aerobic synergistic purification: The photosynthetic bacterial flora utilizes light energy to drive metabolism, simultaneously assimilating ammonia nitrogen, hydrogen sulfide and other pollutants in the anaerobic zone of the sediment, making up for the spatial limitations of the nitrifying bacterial flora, and forming a global water quality purification network.

[0060] Parameter optimization basis: the proportion is set to 15% to 25%. The proportion of 15% to 25% can adapt to the metabolic demand of high-density breeding. When the proportion is lower than 15%, the toxin conversion flux is insufficient, and the accumulation risk of nitrite increases; higher than 25% indicates that the ammonia nitrogen generated in module one exceeds the standard, or occupies the resources of module three, resulting in imbalance of system economy and functional balance.

[0061] Module three: host health oriented gain and quality improvement module flora, accounting for 15% to 25% of the total number of live bacteria.

[0062] Composition: composed of Lactobacillus plantarum and Bacillus amyloliquefaciens FZB42 in a ratio of 1:1.

[0063] Action and mechanism:

[0064] (1) Targeted delivery pathway: the strains in this module are sprayed into the water body together with modules one and two, are wrapped and enriched in the biological floc by the EPS generated by module one. The aquaculture animals actively ingest these biological flocs, so that the probiotics in this module can enter the intestinal tract at high concentration. Lactobacillus plantarum optimizes the intestinal environment: a large amount of lactic acid is produced in the intestinal tract, the pH value of the intestinal tract is reduced, an acidic environment that is not conducive to the growth of pathogenic bacteria such as Vibrio is formed, and an advantage for the colonization of Bacillus amyloliquefaciens FZB42 is provided.

[0065] (2) Intestinal microecological regulation and multi-target bacteriostatic-immune activation: Lactobacillus plantarum metabolizes and produces acid to inhibit the proliferation of pathogenic bacteria, while providing a colonization advantage for Bacillus amyloliquefaciens FZB42. FZB42 directly antagonizes pathogenic bacteria by secreting bacillomycin, bacillolysin and other broad-spectrum antibacterial substances, and its cell wall components (peptidoglycan, etc.) can activate the host's non-specific immune response (such as the increase of lysozyme activity), and construct a pathogenic defense barrier.

[0066] Parameter optimization basis: 15% to 25% ensures the intestinal colonization concentration threshold. Lower than 15% it is difficult to form a dominant flora, and the immune gain effect is weakened; higher than 25% leads to the imbalance of water purification module proportion, increases the risk of environmental stress, and violates the principle of "internal stability and external purification" of the collaborative design.

[0067] Example 1:

[0068] A composite flora for improving water quality and water product quality in ecological aquaculture, accounting for the total number of live bacteria,

[0069] consisting of 50% of environmental purification and biological flocculation carrier module flora, 25% of pollutant deep conversion and water quality stabilization module flora, and 25% of host health oriented gain and quality improvement module flora;

[0070] The environmental purification and bioflocculation carrier module flora is composed of equal proportions of Bacillus subtilis and Bacillus licheniformis;

[0071] The pollutant deep conversion and water quality stabilization module flora is composed of equal proportions of nitrifying bacteria and photosynthetic bacteria;

[0072] The host health directional gain and quality improvement module flora is composed of equal proportions of Lactobacillus plantarum and Bacillus amyloliquefaciens.

[0073] The nitrifying bacteria are obtained by mixing equal proportions of Nitrosomonas and Nitrobacter;

[0074] The photosynthetic bacteria are obtained by mixing equal proportions of Rhodopseudomonas, purple sulfur bacteria, green sulfur bacteria, cyanobacteria and halobacteria.

[0075] The Bacillus amyloliquefaciens is Bacillus amyloliquefaciens FZB42.

[0076] A composite bacterial liquid for improving the water quality and water product quality of ecological aquaculture, the composite bacterial liquid comprising the composite bacterial flora.

[0077] A preparation method of a composite bacterial liquid for improving the water quality and water product quality of ecological aquaculture, comprising the following steps:

[0078] S1, respectively, the environmental purification and bioflocculation carrier module flora, pollutant deep conversion and water quality stabilization module flora and host health directional gain and quality improvement module flora are subjected to liquid fermentation culture, and the environmental purification and bioflocculation carrier module flora mother liquor, pollutant deep conversion and water quality stabilization module flora mother liquor and host health directional gain and quality improvement module flora mother liquor are obtained;

[0079] S2, the environmental purification and bioflocculation carrier module flora mother liquor, pollutant deep conversion and water quality stabilization module flora mother liquor and host health directional gain and quality improvement module flora mother liquor are mixed according to the corresponding proportion, and the composite bacterial liquid is obtained.

[0080] The viable bacterial concentration in the environmental purification and bioflocculation carrier module flora mother liquor, pollutant deep conversion and water quality stabilization module flora mother liquor and host health directional gain and quality improvement module flora mother liquor is not less than 1×10 10 CFU / ml.

[0081] Example 2:

[0082] A composite bacterial flora for improving the water quality and water product quality of ecological aquaculture, accounting for the total number of viable bacteria,

[0083] 70% of the environmental purification and bio-flocculation carrier module flora, 15% of the pollutant deep conversion and water quality stabilization module flora, and 15% of the host health directional gain and quality improvement module flora;

[0084] The environmental purification and bio-flocculation carrier module flora is composed of equal proportions of Bacillus subtilis and Bacillus licheniformis;

[0085] The pollutant deep conversion and water quality stabilization module flora is composed of equal proportions of nitrifying bacteria and photosynthetic bacteria;

[0086] The host health directional gain and quality improvement module flora is composed of equal proportions of Lactobacillus plantarum and Bacillus amyloliquefaciens.

[0087] The nitrifying bacteria are obtained by mixing equal proportions of Nitrosomonas and Nitrobacter;

[0088] The photosynthetic bacteria are obtained by mixing equal proportions of Rhodopseudomonas, purple sulfur bacteria, green sulfur bacteria, cyanobacteria, and halobacteria.

[0089] The Bacillus amyloliquefaciens is Bacillus amyloliquefaciens FZB42.

[0090] A composite bacterial liquid for improving the water quality and water product quality of ecological aquaculture, the composite bacterial liquid comprising the composite bacterial flora.

[0091] A preparation method of a composite bacterial liquid for improving the water quality and water product quality of ecological aquaculture, comprising the following steps:

[0092] S1, respectively, liquid fermentation culture of environmental purification and bio-flocculation carrier module flora, pollutant deep conversion and water quality stabilization module flora and host health directional gain and quality improvement module flora, respectively, to obtain environmental purification and bio-flocculation carrier module flora mother liquor, pollutant deep conversion and water quality stabilization module flora mother liquor and host health directional gain and quality improvement module flora mother liquor;

[0093] S2, according to the corresponding proportion, the environmental purification and bio-flocculation carrier module flora mother liquor, the pollutant deep conversion and water quality stabilization module flora mother liquor and the host health directional gain and quality improvement module flora mother liquor are mixed to obtain the composite bacterial liquid.

[0094] The viable bacterial concentration in the environmental purification and bio-flocculation carrier module flora mother liquor, the pollutant deep conversion and water quality stabilization module flora mother liquor, and the host health directional gain and quality improvement module flora mother liquor is not less than 1x10 10 CFU / ml.

[0095] Example 3:

[0096] A complex microbial community for improving water quality and aquatic product quality in ecological aquaculture, based on the percentage of total viable bacteria.

[0097] It consists of 60% environmental purification and bioflocculation carrier module microbial community, 20% pollutant deep transformation and water quality stabilization module microbial community, and 20% host health-oriented enhancement and quality improvement module microbial community.

[0098] The microbial community of the environmental purification and bioflocculation carrier module is composed of Bacillus subtilis and Bacillus licheniformis in equal proportions.

[0099] The microbial community of the pollutant deep transformation and water quality stabilization module is composed of nitrifying bacteria and photosynthetic bacteria in equal proportions.

[0100] The host health-oriented gain and quality enhancement module microbial community is composed of Lactobacillus plantarum and Bacillus amyloliquefaciens in equal proportions.

[0101] The nitrifying bacteria group is obtained by mixing Nitrosomonas and Nitrobacterium in equal proportions.

[0102] The photosynthetic bacterial community was obtained by mixing Rhodopseudomonas, purple sulfur bacteria, green sulfur bacteria, cyanobacteria, and halobacteria in equal proportions.

[0103] The amyloliquefaciens mentioned is amyloliquefaciens FZB42.

[0104] A compound bacterial solution for improving water quality and aquatic product quality in ecological aquaculture, the compound bacterial solution containing the aforementioned compound bacterial group.

[0105] A method for preparing a compound bacterial solution for improving water quality and aquatic product quality in ecological aquaculture includes the following steps:

[0106] S1. Liquid fermentation culture was carried out on the bacterial communities of the environmental purification and bioflocculation carrier module, the bacterial communities of the pollutant deep transformation and water quality stabilization module, and the bacterial communities of the host health directional gain and quality improvement module, respectively, to obtain the mother liquor of the bacterial communities of the environmental purification and bioflocculation carrier module, the mother liquor of the bacterial communities of the pollutant deep transformation and water quality stabilization module, and the mother liquor of the bacterial communities of the host health directional gain and quality improvement module, respectively.

[0107] S2. Mix the mother liquor of the environmental purification and bioflocculation carrier module, the mother liquor of the pollutant deep transformation and water quality stabilization module, and the mother liquor of the host health directional enhancement and quality improvement module in the appropriate proportions to obtain a composite bacterial solution.

[0108] The viable bacteria concentration in the bacterial culture mother liquor of the environmental purification and bioflocculation carrier module, the bacterial culture mother liquor of the pollutant deep transformation and water quality stabilization module, and the bacterial culture mother liquor of the host health-oriented enhancement and quality improvement module is not less than 1×10⁻⁶. 10 CFU / ml.

[0109] Experimental Example 1: Verify the high efficiency of probiotic delivery, break through the technical barrier

[0110] The composite bacteria liquid for improving water quality and aquatic product quality prepared in Example 3 was selected as the experimental group.

[0111] Through the grass carp breeding experiment and intestinal flora detection, compared with the control group in which the module three strains were directly sprayed into the water, the experimental group using the composition of the application achieved a breakthrough in the efficiency of probiotic delivery.

[0112] The specific data are as follows: after 72 hours of application, the intestinal microorganisms of grass carp were collected for quantitative culture counting of target strains (Lactobacillus plantarum).

[0113] As shown in Figure 1 , it is a local schematic diagram of Lactobacillus plantarum colony plate.

[0114] As shown in Figure 2 , it is a schematic diagram of the gram staining microscope photo of the colony plate.

[0115] The above Figure 1 combined Figure 2 The results show that the Lactobacillus plantarum in the intestinal tract of grass carp is successfully cultured, and the quantitative detection results are shown in Table 1.

[0116] In the intestinal tract of the control group of grass carp, the colonization number of Lactobacillus plantarum was 1.5×10 4 CFU / g, which was difficult to form a dominant population; while in the experimental group of grass carp using the application, the colonization number of Lactobacillus plantarum in the intestinal tract was as high as 2.8×10 7 CFU / g. The colonization number of the experimental group was more than three orders of magnitude (about 1800 times) higher than that of the control group, and had reached the dominant population level of exerting significant physiological effects in the intestinal tract.

[0117] This fully proves that the "in-situ bioflocculation delivery" technology of the application can overcome the dilution effect of a large water body, and realize efficient and targeted delivery of probiotics to the intestinal tract of the host.

[0118] Table 1 Quantitative culture counting of Lactobacillus plantarum in the intestinal tract of grass carp

[0119] Group Average CFU / g (x 10 4 )]]> Standard deviation Fold difference (experimental / control) p value Control group 1.5 ±0.3 1 — Experimental group 2800 ±420 -1867 <0.001

[0120] Experimental Example 2: Verify that the water quality regulation and the disease resistance and growth promotion effects are synergistically enhanced

[0121] 2.1 In a 60-day grass carp breeding experiment (initial average weight 100 g / tail), the application showed significant systemic advantages.

[0122] Table 2 Growth index of grass carp and water quality adjustment in 60-day grass carp culture experiment

[0123]

[0124] Table 2 shows that in terms of growth and survival performance, the survival rate of grass carp in the experimental group (using the present application) was as high as 96% at the end of the culture, and the average weight gain rate reached 195% (final weight about 295g). The control group (using only the single water purification bacteria agent of module two): the survival rate was 82%, and the average weight gain rate was only 140% (final weight about 240g).

[0125] The present application improves the survival rate by 14 percentage points, and the weight gain rate is relatively increased by nearly 40%, which shows that the present application has a great promoting effect on improving the survival rate of aquatic products, etc.

[0126] 2.2 In terms of water quality dynamic regulation ability, the water body was detected daily during the entire culture period.

[0127] Table 3 Change of COD and ammonia nitrogen concentration in water during 60-day grass carp culture experiment

[0128]

[0129] Note: The initial concentrations of COD and ammonia nitrogen are simulated high-density culture initial organic load and initial accumulation of residual feed and excretion.

[0130] Table 3 shows that in the experimental group, the average concentration of COD (chemical oxygen demand) in the water body is stably below 25mg / L, and the concentration of ammonia nitrogen (NH4 + -N) is maintained at a safe level of below 0.2mg / L for a long time.

[0131] This shows that the composition has high dynamic degradation ability for continuously generated pollutants. In the control group, the average concentration of COD in the water body fluctuates around 45mg / L, and the concentration of ammonia nitrogen frequently breaks through 0.6mg / L, which constitutes a continuous stress to the cultured animals.

[0132] The results show that compared with professional water purification bacteria agents, the present application can reduce the concentrations of COD and ammonia nitrogen by more than 50% and 65%. This multi-module substrate relay cooperative mechanism breaks through the efficiency attenuation bottleneck of traditional single bacteria agents under complex organic load conditions, and provides an innovative solution for high-density culture water quality regulation.

[0133] Finally, it should be noted that the above description is only a preferred embodiment of the present application and is not used to limit the protection scope of the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. A composite bacterial flora for improving water quality-aquaculture product quality for ecological aquaculture, characterized by, By total live bacteria count, The environmental purification and bioflocculation carrier module bacterial population is composed of equal proportions of Bacillus subtilis and Bacillus licheniformis; The pollutant deep conversion and water quality stabilization module bacterial population is composed of equal proportions of nitrifying bacteria and photosynthetic bacteria; The host health directional gain and quality improvement module bacterial population is composed of equal proportions of Lactobacillus plantarum and Bacillus amyloliquefaciens. By total live bacteria count, 2. The composite microbial flora for improving water quality and water product quality of ecological aquaculture according to claim 1, characterized in that, The environmental purification and bioflocculation carrier module bacterial population is composed of 60% of the total, the pollutant deep conversion and water quality stabilization module bacterial population is composed of 20% of the total, and the host health directional gain and quality improvement module bacterial population is composed of 20% of the total. The nitrifying bacteria are obtained by mixing equal proportions of Nitrosomonas and Nitrobacter; 3. The composite microbial flora for improving water quality and aquatic product quality of ecological aquaculture according to claim 1 or 2, characterized in that, The photosynthetic bacteria are obtained by mixing equal proportions of Rhodopseudomonas, purple sulfur bacteria, green sulfur bacteria, cyanobacteria, and halobacteria. The Bacillus amyloliquefaciens is Bacillus amyloliquefaciens FZB42.

4. The complex microbial flora for improving water quality and aquatic product quality of ecological aquaculture according to claim 1 or 2, characterized in that, The complex bacterial solution comprises the complex bacterial population according to any one of claims 1-4.

5. A composite bacterial liquid for improving water quality and aquatic product quality in ecological aquaculture, characterized in that, The method comprises the following steps:

6. The preparation method of the complex bacterial liquid for improving water quality and aquatic product quality of ecological aquaculture according to claim 5, characterized in that, S1, respectively, the environmental purification and bioflocculation carrier module bacterial population, pollutant deep conversion and water quality stabilization module bacterial population and host health directional gain and quality improvement module bacterial population are subjected to liquid fermentation culture, and the environmental purification and bioflocculation carrier module bacterial population mother liquor, the pollutant deep conversion and water quality stabilization module bacterial population mother liquor and the host health directional gain and quality improvement module bacterial population mother liquor are obtained; S2, the environmental purification and bioflocculation carrier module bacterial population mother liquor, the pollutant deep conversion and water quality stabilization module bacterial population mother liquor and the host health directional gain and quality improvement module bacterial population mother liquor are mixed according to the corresponding proportion, and the complex bacterial solution is obtained. The complex bacterial solution is directly and uniformly sprayed on the surface of the aquaculture water after being embedded in the hydrogel microcapsule.

7. The method according to claim 6, wherein the method is characterized by, The active bacteria concentration in the environmental purification and bioflocculation carrier module, the pollutant deep conversion and water quality stabilization module, and the host health directional gain and quality improvement module is not less than 1 x 10 10 CFU / ml.

8. The use of the complex bacterial liquid for improving water quality and aquatic product quality of ecological aquaculture according to claim 5, characterized in that, Based on the first use or water quality deterioration of the aquaculture water, 1L of the complex bacterial solution is used for 1000 cubic meters of water. 9.The use method of the compound bacteria liquid for improving water quality and aquatic product quality of ecological aquaculture according to claim 8, characterized in that, Based on the daily maintenance of the aquaculture water, 0.5L of the complex bacterial solution is used for 1000 cubic meters of water every 10-15 days.

10. The use of the compound microbial solution for improving water quality and aquatic product quality of ecological aquaculture according to claim 8, characterized in that, ​

Images