CN109626592B - Method for adjusting water quality of eel breeding pond in net cage - Google Patents

Method for adjusting water quality of eel breeding pond in net cage Download PDF

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CN109626592B
CN109626592B CN201811606983.8A CN201811606983A CN109626592B CN 109626592 B CN109626592 B CN 109626592B CN 201811606983 A CN201811606983 A CN 201811606983A CN 109626592 B CN109626592 B CN 109626592B
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chitosan
water quality
shell material
water
shells
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CN109626592A (en
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鲁敏
陈晋
杨潇
景斐
张建设
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Zhejiang Ocean University ZJOU
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    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00Biological treatment of water, waste water, or sewage
    • C02F3/34Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G22/00Cultivation of specific crops or plants not otherwise provided for
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01KANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K61/00Culture of aquatic animals
    • A01K61/10Culture of aquatic animals of fish
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00Biological treatment of water, waste water, or sewage
    • C02F3/34Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • C02F3/347Use of yeasts or fungi
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00Biological treatment of water, waste water, or sewage
    • C02F3/34Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • C02F3/348Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the way or the form in which the microorganisms are added or dosed
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00Nature of the contaminant
    • C02F2101/10Inorganic compounds
    • C02F2101/20Heavy metals or heavy metal compounds
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2103/00Nature of the water, waste water, sewage or sludge to be treated
    • C02F2103/20Nature of the water, waste water, sewage or sludge to be treated from animal husbandry
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02ATECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/80Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
    • Y02A40/81Aquaculture, e.g. of fish

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  • Microbiology (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Engineering & Computer Science (AREA)
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  • Chemical & Material Sciences (AREA)
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Abstract

The invention provides a method for adjusting water quality of a net cage eel breeding pond, which comprises the following steps: periodically using an immobilized probiotic water quality regulator; filter-feeding fishes and/or scrape-feeding fishes are put in the water outside the net cage; periodically transplanting and renewing the submerged plants. The immobilized probiotic water quality regulator comprises a functional shell material modified by chitosan and probiotics carried on the shell material; the specific surface area of the functional shell material before being modified by chitosan is 1.0-2.0 m2(ii)/g, the average pore diameter is 0.5 to 45 nm. The adjusting method provided by the invention comprehensively utilizes the probiotics water quality regulating agent, the released ecological functional fish and the biological treatment, improves the water quality of the pool for culturing the eels in the net cage, improves the water body utilization rate, and obtains the fishes and plants for water transfer, thereby improving the economic benefit; the water quality regulator has good dispersibility, multiple modification sites, strong adsorption capacity and water retention performance, high immobilization degree on probiotics, and can keep high activity of the probiotics and prolong the storage time.

Description

Method for adjusting water quality of eel breeding pond in net cage
Technical Field
The invention belongs to the technical field of aquaculture, and particularly relates to a method for adjusting water quality of a pool for culturing eels in a net cage.
Background
Monopteri albi, commonly called eel, belongs to Ostertagia, order Hegilles, family Hegillidae, genus Monopteri albi, and is mainly distributed in China, Vietnam, Korea, Thailand, Japan, and is the main famous and high-quality fresh water product in China. Eel is one of famous and high-quality fishes which are cultured and developed rapidly in recent years in China, and the main culture production areas of eel are distributed in the provinces of Hubei, Hunan, Anhui, Jiangxi, Zhejiang and the like in the middle and lower reaches of Yangtze river. Eel farming has become one of the leading industries of the aquaculture industry in these areas.
For a long time, the artificial breeding method of the finless eels mainly comprises three methods of pond breeding, paddy field breeding and net cage breeding; in recent years, the culture area of finless eels is enlarged year by year, and the culture mode mainly focuses on cage culture except for small part of rice field culture. The method for culturing the finless eels by arranging the net cages in the pond is a novel culture technology, is also a novel mode for culturing the finless eels, and has the advantages of small water surface occupation, low fixed asset investment, easy culture, easy capture, convenient management, convenient operation, low labor intensity, high bait utilization rate, quick growth of the finless eels, high yield, short culture period, quick response, high unit area benefit, easy acceptance by farmers, easy popularization and the like.
The net cage culture of the finless eels is the main culture mode of the finless eels in China at present, and the culture mode has the following defects: (1) the feed for feeding the finless eels is high-protein compound feed and minced fillet, and the water quality is greatly polluted by the finless eel excrement and residual feed; (2) the mesh is small, generally 30 meshes/square centimeter, and is easy to be blocked by sludge, attached algae and finless eel excrement, the exchange of water inside and outside the net cage is poor, and the water quality in the net cage is easy to deteriorate; (3) poor water quality, high incidence rate of finless eels, slow growth, large feed coefficient, low benefit and large breeding risk.
The water quality regulation of the finless eels cultured in the net cage is very important, and the success or failure of the culture is determined. The water quality regulation of the finless eels cultured in the traditional net cage mainly comprises water changing and quicklime splashing, 1) the water changing pollutes the environment, the finless eels are easy to cause stress reaction when the water changing exceeds 1/3 once, and the water source is difficult to be carried out in a poor place; 2) the finless eel likes an acidic environment, and the quicklime water is alkaline, so that the consumption is large or the quicklime water is frequently used, and the growth of the finless eel is influenced.
The prior art is to add chemical oxygen release agent, beneficial microorganism, disinfectant, disease-resistant agent and the like into the water body, and although certain progress is made, the method also has a plurality of defects, wherein the ecological purification method added with the beneficial microorganism can achieve the purpose of regulation more effectively and for a long time. If the foraging activity of filter-feeding or scrape-feeding fishes is utilized, phytoplankton and residual baits are ingested, the exchange and circulation of water bodies inside and outside the net cage are facilitated, and the water bodies are purified; the submerged plants take nutritive salt, illumination and growth space in the water body as growth resources to absorb nitrogen and phosphorus in the water body; the microbial preparation can purify water quality, reduce diseases and directly or indirectly provide basic bait for cultured organisms. Therefore, the ecological purification method is adopted to regulate the water quality of the eel pond in the net cage, has the characteristics of low consumption, high efficiency and environmental safety, and is a good method which is effective, protects the environment and avoids secondary pollution.
Disclosure of Invention
The invention aims to provide the immobilized probiotic water quality regulator which has good dispersibility, more modification sites, strong adsorption capacity and water retention performance, high immobilization degree on probiotics, prolonged preservation time and maintained high activity of the probiotics.
The invention also aims to provide a method for regulating the water quality of the net cage eel breeding pond, which comprehensively utilizes the probiotics water quality regulating agent, puts in a suitable place for breeding ecological functional fishes, treats organisms, improves the water quality of the net cage eel breeding pond and improves the water utilization rate.
The technical scheme adopted by the invention for realizing the purpose is as follows:
the water quality regulating agent comprises a functional shell material modified by chitosan and probiotics carried on the shell material; and the specific surface area of the functional shell material before being modified by chitosan is 1.0-2.0 m2(ii)/g, the average pore diameter is 0.5 to 45 nm. The water quality regulating agent can degrade harmful substances such as ammonia nitrogen, nitrite and hydrogen sulfide in water by using probiotics, can remove heavy metal ions in the water by using functional shell materials through physical adsorption and ion exchange, reduces water pollution and improves water quality, and meanwhile, the probiotics can also be used as basic bait or feed additive for animals to eat, reduces bait feeding, and achieves the purposes of waste utilization, ecological balance and healthy culture.
Preferably, the functional shell material is subjected to calcination and water modification pretreatment before being modified by chitosan; the calcining temperature is 500-600 ℃, the time is 1-2 h, and the calcining time is in an oxygen-free environment; the temperature of the water is changed to be 150-180 ℃, the time is 12-24 hours, and the reaction kettle is a polytetrafluoroethylene reaction kettle. After the waste shells are calcined and puffed at high temperature and the like, an irregular porous structure is formed, the specific surface area and the pores are increased, and internal micropores and mesopores are puffed, so that the adsorbent has stronger thermal stability and adsorption capacity, is an excellent carrier and adsorbent, and has excellent adsorption and sustained release effects.
Further preferably, the pretreatment step is: cleaning shells, soaking the shells in 0.1-0.5% hydrochloric acid solution for 10-12 h, then placing the shells in a tubular furnace at 500-600 ℃ for anaerobic calcination for 1-2 h, taking out the shells and cooling the shells, crushing the shells and passing through a 100-120-mesh sieve, then adding 5-10 times of 1-2% potassium carbonate solution into the shells, ultrasonically dissolving the shells for 20-30 min under 500-600W, then sending the mixed solution into a polytetrafluoroethylene reaction kettle, reacting the mixed solution for 12-24 h at 150-180 ℃, centrifuging the solution, washing precipitates to be neutral, drying the precipitates for 6-8 h at 60-80 ℃, and obtaining the shells for later use.
Preferably, the chitosan-modified functional shell material is prepared by the following steps: adding the pretreated shell material into a chitosan solution, carrying out mixed ultrasonic treatment for 15-30 min under the power of 500-800W, and carrying out vacuum drying for 12-24 h at the temperature of 70-90 ℃ to obtain the chitosan/chitosan composite material; the chitosan solution contains kojic acid and sodium naphthenate. The surfaces of the probiotics are electronegative, and the shell powder material with the hydrophilic characteristic is difficult to combine with the probiotics under the action of water molecule acting force and hydrogen bonds, so that the shell material is modified by chitosan, and the probiotics can be fixed on the shell material through ion interaction and adsorption flocculation.
Further preferably, the concentration of the chitosan solution is 1-2 mg/L, and the weight ratio of chitosan to the shell material is 1: 2-3 in terms of chitosan; the contents of kojic acid and sodium naphthenate in the chitosan solution are 0.12-0.32 wt% and 0.15-0.5 wt%, respectively. Kojic acid and sodium naphthenate are combined by similar chemical bonds between hydrophilic group carboxyl in molecules and calcium carbonate, so that the surface energy of calcium carbonate molecules is reduced, the mutual attraction among calcium carbonate molecules is weakened, the agglomeration phenomenon among shell powder is improved, the dispersibility of the shell powder is enhanced, and meanwhile, functional groups on the surface of chitosan can be activated, so that a layer of colloid is formed on the surface of the shell material modified by the chitosan, the water-retaining property of the shell material is enhanced, the immobilization degree of the shell material on probiotics is improved, the storage time of immobilized probiotic products is prolonged, and the high activity of the probiotics is maintained.
Still further preferably, the preparation step of the chitosan solution is as follows: adding 5-10 times of 1-2% hydrochloric acid solution into chitosan, continuously stirring and dissolving, and then adding kojic acid, sodium naphthenate and distilled water to prepare a solution with the concentration of 1-2 mg/L in terms of chitosan.
Preferably, the immobilized probiotic is prepared by the following steps: adding 2-5 wt% of tea polyphenol into the activated and expanded probiotic culture solution, adding a chitosan modified functional shell material according to the proportion of 1: 0.5-1, magnetically stirring for 20-30 min, adding sodium alginate and 1: 0.3-0.5 of glycerol in the weight ratio of the sodium alginate to the shell material, uniformly mixing, standing for 1-2 h, and removing the upper clear water to obtain the probiotic culture solution. The shell material modified by chitosan has the characteristics of good adsorption performance and rich raw materials, and the characteristic of high-efficiency enrichment of nitrogen, phosphorus and heavy metals of probiotics are combined, so that the shell material can quickly adsorb pollutants in water, and simultaneously consumes nutrient substances such as nitrogen, phosphorus and the like in the wastewater through the growth and metabolism of the probiotics, thereby achieving the effect of deeply purifying the water body.
Preferably, the culture medium for probiotic culture is prepared according to the formula of an MS culture medium, the culture temperature is 25-45 ℃, and the propagation time is 2-8 days.
Preferably, the probiotic bacteria are selected from at least two of photosynthetic bacteria, bacillus subtilis, yeast, EM bacteria and lactic acid bacteria.
The invention also discloses a method for adjusting the water quality of the eel breeding pond in the net cage, which comprises the following steps:
a, periodically using an immobilized probiotic water quality regulator;
b, stocking filter-feeding fishes and/or scrape-feeding fishes in the water outside the net cage;
and c, periodically transplanting and updating the submerged plants. The regulation method comprehensively utilizes the probiotic water quality regulation agent, the fish with the stocking ecological function and the ecological regulation method for biological treatment, supplements each other, exerts the whole functions, improves the water quality, the water body utilization rate and the water body eutrophication state, and can directly or indirectly improve the economic benefit through the processing of the cultured animals and plants.
Preferably, the use period of the immobilized probiotic water quality regulator is 1-2 months/time, and the use amount is 200-500 mg/m3A net cage.
Preferably, the filter-feeding fishes are silver carps and bighead carps, and the scraper-feeding fishes are plagiognathops microlepis; the initial stocking density of the fishes is 10-20 tails/m3The average weight is 50-150 g. The foraging activity of the fishes in the pond is beneficial to the exchange and circulation of water bodies inside and outside the net cage, filter-feeding fishes ingest phytoplankton, the water bodies are purified, scraping-feeding fishes ingest filamentous algae and the like, the box body is purified, meanwhile, the fishes ingest the residual bait of the edible residues, and the bottom of the pond is purified.
Preferably, the period for transplanting and updating the submerged plants is 45-60 days; the submerged plant is at least one selected from herba Swertiae Dilutae, Goldfish algae, Foliumet Sedum, and hydrilla verticillata. The submerged plant takes nutritive salt, illumination and growth space in the water body as growth resources, absorbs nitrogen and phosphorus in the water body, forms a competitive relationship with algae simultaneously, plays a role in inhibiting the growth of the algae, can effectively inhibit allelochemicals secreted by the submerged plant, and is suitable for ecological management and regulation of the aquaculture water body.
The invention has the beneficial effects that:
1) according to the invention, the shell material modified by chitosan is used as a carrier of probiotics, the shell material is not agglomerated, has good dispersibility, more modification sites, strong adsorption capacity and water retention performance and high immobilization degree on the probiotics, so that the storage time of the immobilized probiotics is prolonged, the high activity of the probiotics is maintained, and the effects of deeply purifying water and improving dissolved oxygen are achieved;
2) the water quality regulating agent of the immobilized probiotics can degrade harmful substances such as ammonia nitrogen, nitrite and hydrogen sulfide in water, can also remove heavy metal ions in the water by physical adsorption and ion exchange by using functional shell materials, improves water quality, can be used as basic bait or feed additive for animals to eat, reduces bait feeding, and achieves the purposes of waste utilization, ecological balance and healthy culture;
3) the ecological regulation method comprehensively utilizes the probiotic water quality control agent, the released ecological functional fish and the biological treatment, plays a synergistic role, improves the water quality of the eel breeding pond in the net cage, improves the water body utilization rate, improves the water body eutrophication state, and can directly or indirectly improve the economic benefit through the harvest and processing of the bred animals and plants.
The invention adopts the technical scheme to provide the method for adjusting the water quality of the eel breeding pond in the net cage, overcomes the defects of the prior art, and has reasonable design and convenient operation.
Drawings
FIG. 1 is a distribution diagram of the adsorption pore size of a shell material before modification with chitosan prepared in example 3;
FIG. 2 is a scanning electron micrograph of a shell material before pretreatment in example 3;
FIG. 3 is a scanning electron microscope image of the shell material before modification with chitosan prepared in example 3.
Detailed Description
The technical solution of the present invention is further described in detail below with reference to the following detailed description and the accompanying drawings:
the photosynthetic bacteria and Bacillus subtilis used in the present invention and examples were obtained from Yufeng Biotechnology Ltd, Nanjing, Lactobacillus was obtained from Jiayi bioengineering Ltd, Shandong, middle-jiao Yihong bioengineering Ltd, EM bacteria and yeast were obtained from Yihong Biotechnology Ltd, Cangzhou.
Example 1:
the water quality regulating agent comprises a functional shell material modified by chitosan and probiotics carried on the shell material. The water quality regulating agent can degrade harmful substances such as ammonia nitrogen, nitrite and hydrogen sulfide in water by using probiotics, can remove heavy metal ions in the water by using functional shell materials through physical adsorption and ion exchange, reduces water pollution and improves water quality, and meanwhile, the probiotics can also be used as basic bait or feed additive for animals to eat, reduces bait feeding, and achieves the purposes of waste utilization, ecological balance and healthy culture.
The functional shell material is calcined and pretreated by water modification before being modified by chitosan; the calcining temperature is 500 ℃, the time is 2 hours, and the environment is oxygen-free; the temperature of the water is changed to 150 ℃, the time is 24 hours, and the reaction kettle is a polytetrafluoroethylene reaction kettle. After the waste shells are calcined and puffed at high temperature and the like, an irregular porous structure is formed, the specific surface area and the pores are increased, and internal micropores and mesopores are puffed, so that the adsorbent has stronger thermal stability and adsorption capacity, is an excellent carrier and adsorbent, and has excellent adsorption and sustained release effects.
The pretreatment steps are as follows: cleaning shells, soaking the shells in 0.1% hydrochloric acid solution for 12h, then placing the shells in a tube furnace at 500 ℃ for anaerobic calcination for 2h, taking out the shells, cooling the shells, crushing the shells, sieving the shells with a 100-mesh sieve, adding 1% potassium carbonate solution in an amount which is 5 times the amount of the shells, ultrasonically dissolving the shells for 30min under 500W power, sending the mixed solution into a polytetrafluoroethylene reaction kettle, reacting the mixed solution for 24h at 150 ℃, centrifuging the solution, washing precipitates to be neutral, and drying the precipitates for 8h at 60 ℃ to obtain the shell powder for later use.
The functional shell material modified by chitosan is prepared by the following steps: adding the pretreated shell material into a chitosan solution, carrying out mixed ultrasonic treatment for 30min under 500 power, and carrying out vacuum drying for 24h at 70 ℃ to obtain the chitosan/chitosan composite material; the chitosan solution contains kojic acid and sodium naphthenate. The surfaces of the probiotics are electronegative, and the shell powder material with the hydrophilic characteristic is difficult to combine with the probiotics under the action of water molecule acting force and hydrogen bonds, so that the shell material is modified by chitosan, and the probiotics can be fixed on the shell material through ion interaction and adsorption flocculation.
The concentration of the chitosan solution is 1mg/L, and the weight ratio of the chitosan to the shell material is 1:2 in terms of chitosan; the contents of kojic acid and sodium naphthenate in the chitosan solution were 0.15 wt% and 0.15 wt%, respectively. Kojic acid and sodium naphthenate are combined by similar chemical bonds between hydrophilic group carboxyl in molecules and calcium carbonate, so that the surface energy of calcium carbonate molecules is reduced, the mutual attraction among calcium carbonate molecules is weakened, the agglomeration phenomenon among shell powder is improved, the dispersibility of the shell powder is enhanced, and meanwhile, functional groups on the surface of chitosan can be activated, so that a layer of colloid is formed on the surface of the shell material modified by the chitosan, the water-retaining property of the shell material is enhanced, the immobilization degree of the shell material on probiotics is improved, the storage time of immobilized probiotic products is prolonged, and the high activity of the probiotics is maintained.
The preparation method of the chitosan solution comprises the following steps: adding 5 times of 1% hydrochloric acid solution into chitosan, stirring to dissolve, adding kojic acid, sodium naphthenate and distilled water, and making into 1mg/L solution.
The immobilized probiotics are prepared by the following steps: adding 2% by weight of tea polyphenol into the activated and expanded probiotic culture solution, adding a chitosan-modified functional shell material according to the proportion of 1:0.5, magnetically stirring for 30min, adding sodium alginate and 1:0.3 glycerin according to the weight ratio of 1:0.3 of the shell material, uniformly mixing, standing for 2h, and removing the upper clear water to obtain the probiotic culture solution. The shell material modified by chitosan has the characteristics of good adsorption performance and rich raw materials, and the characteristic of high-efficiency enrichment of nitrogen, phosphorus and heavy metals of probiotics are combined, so that the shell material can quickly adsorb pollutants in water, and simultaneously consumes nutrient substances such as nitrogen, phosphorus and the like in the wastewater through the growth and metabolism of the probiotics, thereby achieving the effect of deeply purifying the water body.
The culture medium for probiotic culture is prepared according to the formula of an MS culture medium, the culture temperature is 28 ℃, and the propagation time is 8 days.
The probiotic bacteria are selected from at least two of photosynthetic bacteria, Bacillus subtilis, yeast, EM bacteria and lactic acid bacteria. In this example, Bacillus subtilis, EM and lactic acid bacteria were selected.
A method for adjusting the water quality of a pool for culturing eels in a net cage comprises the following steps:
a, periodically using an immobilized probiotic water quality regulator;
b, stocking filter-feeding fishes and/or scrape-feeding fishes in the water outside the net cage;
and c, periodically transplanting and updating the submerged plants. The regulation method comprehensively utilizes the probiotic water quality regulation agent, the fish with the stocking ecological function and the ecological regulation method for biological treatment, supplements each other, exerts the whole functions, improves the water quality, the water body utilization rate and the water body eutrophication state, and can directly or indirectly improve the economic benefit through the processing of the cultured animals and plants.
The use period of the immobilized probiotic water quality control agent is 1 month/time, and the use amount is 200mg/m3A net cage.
The filter-feeding fish is silver carp and bighead carp, and the scraper-feeding fish is plagiognathops microlepis; the initial stocking density of the fish is 10 fish/m3The average body weight was 150 g. The foraging activity of the fishes in the pond is beneficial to the exchange and circulation of water bodies inside and outside the net cage, filter-feeding fishes ingest phytoplankton, the water bodies are purified, scraping-feeding fishes ingest filamentous algae and the like, the box body is purified, meanwhile, the fishes ingest the residual bait of the edible residues, and the bottom of the pond is purified.
The period for transplanting and updating the submerged plants is 45 days; the submerged plant is at least one selected from the group consisting of tape grass, Goldfish algae, Foliumet-tailed algae, and hydrilla verticillata, in this example, Goldfish algae is selected. The submerged plant takes nutritive salt, illumination and growth space in the water body as growth resources, absorbs nitrogen and phosphorus in the water body, forms a competitive relationship with algae simultaneously, plays a role in inhibiting the growth of the algae, can effectively inhibit allelochemicals secreted by the submerged plant, and is suitable for ecological management and regulation of the aquaculture water body.
Example 2:
the method for adjusting the water quality of the eel breeding pond in the net cage specifically comprises the following steps:
1) the immobilized probiotic water quality regulator is used periodically and is used once every 2 months, and the using amount is 350mg/m3A net cage;
2) silver carp, bighead carp and plagiognathops microlepis are released in water outside the net cage, and the initial release density is 20 fish/m3Average body weight of 60 g;
3) transplanting and renewing the tape grass and the hornwort periodically, wherein the transplanting and renewing period is 60 days.
The preparation method of the immobilized probiotic water quality regulator used in the regulation method comprises the following steps:
1) cleaning shells, soaking the shells in 0.5% hydrochloric acid solution for 10 hours, then placing the shells in a tube furnace at the temperature of 600 ℃ for anaerobic calcination for 1 hour, taking out the shells, cooling the shells, crushing the shells, sieving the shells by a 110-mesh sieve, adding 10 times of 2% potassium carbonate solution into the shells, ultrasonically dissolving the shells for 20 minutes under the power of 600W, then sending the mixed solution into a polytetrafluoroethylene reaction kettle, reacting the mixed solution for 12 hours at the temperature of 180 ℃, centrifuging the solution, washing precipitates to be neutral, and drying the precipitates for 6 hours at the temperature of 70 ℃ to obtain the shells for later use;
2) adding 10 times of 2% hydrochloric acid solution into chitosan, stirring continuously to dissolve, adding kojic acid, sodium naphthenate and distilled water to prepare a solution with a concentration of 2mg/L based on chitosan, wherein the contents of kojic acid and sodium naphthenate are 0.3 wt% and 0.43 wt% respectively;
3) adding the pretreated shell material into a chitosan solution, wherein the weight ratio of chitosan to the shell material is 1:3 calculated by chitosan, mixing and ultrasonically treating for 15min under the power of 800W, and carrying out vacuum drying for 12h at 70 ℃ to obtain a chitosan-modified functional shell material;
4) preparing a culture medium according to an MS culture medium formula, and activating and expanding probiotics at the culture temperature of 40 ℃ for 3 days, wherein the probiotics are photosynthetic bacteria and bacillus subtilis;
5) adding 5% by weight of tea polyphenol into the probiotic culture solution, adding a chitosan modified functional shell material according to the proportion of 1:1, magnetically stirring for 30min, adding sodium alginate and 1:0.5 of glycerol in the weight ratio of the sodium alginate to the shell material of 1:0.8, uniformly mixing, standing for 1h, and removing the upper clear water to obtain the immobilized probiotic.
Example 3:
the method for adjusting the water quality of the eel breeding pond in the net cage specifically comprises the following steps:
1) the immobilized probiotic water quality regulator is used periodically and once every 1.5 months, and the using amount is 450mg/m3A net cage;
2) silver carp and bighead carp are put in the water outside the net cageAnd plagiognathops microlepis, the initial stocking density is 15/m3The average weight is 110 g;
3) transplanting and renewing the tape grass and the hornwort periodically, wherein the transplanting and renewing period is 50 days.
The preparation method of the immobilized probiotic water quality regulator used in the regulation method comprises the following steps:
1) cleaning shells, soaking the shells in 0.2% hydrochloric acid solution for 11h, then placing the shells in a tube furnace at the temperature of 550 ℃ for anaerobic calcination for 1.5h, taking out the shells, cooling the shells, crushing the shells, sieving the crushed shells with a 120-mesh sieve, adding 7 times of 1.5% potassium carbonate solution into the shells, ultrasonically dissolving the shells for 25min under the power of 500W, then sending the mixed solution into a polytetrafluoroethylene reaction kettle, reacting the mixed solution for 18h at the temperature of 165 ℃, centrifuging the solution, washing precipitates to be neutral, and drying the precipitates for 7h at the temperature of 80 ℃ to obtain the shells for later use;
2) adding 8 times of 1.5% hydrochloric acid solution into chitosan, stirring continuously to dissolve, adding kojic acid, sodium naphthenate and distilled water to prepare 1.5mg/L solution calculated by chitosan, wherein the contents of kojic acid and sodium naphthenate are 0.28 wt% and 0.35 wt% respectively;
3) adding the pretreated shell material into a chitosan solution, wherein the weight ratio of chitosan to the shell material is 1:2.5 calculated by chitosan, mixing and ultrasonically treating for 20min under the power of 700W, and vacuum drying for 16h at the temperature of 85 ℃ to obtain the chitosan modified functional shell material;
4) preparing a culture medium according to an MS culture medium formula, and activating and expanding probiotics at the culture temperature of 33 ℃ for 5 days, wherein the probiotics are bacillus subtilis, EM (effective microorganisms) and saccharomycetes;
5) adding 3.5 wt% of tea polyphenol into the probiotic culture solution, adding a chitosan-modified functional shell material according to the proportion of 1:0.85, magnetically stirring for 25min, adding sodium alginate and 1:0.4 glycerin according to the weight ratio of 1:0.6 to the shell material, uniformly mixing, standing for 1.5h, and removing the upper clear water to obtain the immobilized probiotic.
Example 4:
the immobilized probiotic water quality regulating agent optimizes the pretreatment steps of the shell material, and the specific optimization measures are as follows: cleaning shell, soaking in 0.2% hydrochloric acid solution for 11h, calcining in 550 deg.C tube furnace for 1.5h, cooling, pulverizing, sieving with 120 mesh sieve, adding 7 times of 1.5% potassium carbonate solution into shell powder, ultrasonic dissolving at 500W for 25min, reacting at 165 deg.C for 18h, centrifuging, washing precipitate to neutrality, oven drying at 80 deg.C for 7h to obtain shell powder, adding 0.15mM tannic acid and 0.1mM diethyl oxalate into the potassium carbonate solution, adding tannic acid and diethyl oxalate into high temperature water-changing reaction, complexing with calcium ion in shell molecule, and penetrating into pores of shell material, expanding and breaking under ultrasonic impact, therefore, the adsorption capacity of the shell material is enhanced, and meanwhile, the aggregation of functional groups on the surface of calcium carbonate molecules can increase modification sites on the shell material, thereby being beneficial to further modifying the shell material by chitosan.
In the embodiment, the optimization test is performed on the basis of the embodiment 3, and the water quality of the pool for culturing the eels in the net cage is adjusted in the same way as in the embodiment 3 in other steps.
Example 5:
the water quality regulator of immobilized probiotics is prepared by adding kojic acid and sodium naphthenate into chitosan solution during the preparation of chitosan modified functional shell material.
In the embodiment, a comparison test is carried out on the basis of the embodiment 3, and the water quality of the pool for culturing the eels in the net cage is adjusted according to other steps in the embodiment 3.
Example 6:
determination of properties of immobilized probiotic water quality regulator
1) Determining specific surface area and pore size distribution of shell material by NOVA-2200E specific surface area analyzer
A: specific surface area measurement conditions: weighing a sample with the weight of 0.3-0.5 g (taking the bottom of a measuring tube as a standard for covering) for measurement, heating the pretreatment sample on the left side at the heating temperature of 200 ℃, lasting for 30min, waiting for cooling for 10min, placing the measuring tube on a right detection area, adding liquid nitrogen, measuring the Q value by rising the liquid nitrogen, waiting for the Q value difference to be less than 0.001 twice before and after, reducing the liquid nitrogen, filling 80P of air pressure, changing the adsorbate into high-purity nitrogen, and measuring the added liquid nitrogen.
The results of analyzing the shell materials before modification of the chitosan prepared in examples 1 to 5 are shown in table 1 below.
TABLE 1 specific surface area (unit: m) of shell material before chitosan modification2/g)
Example 1 Example 2 Example 3 Example 4 Example 5
Specific surface area 1.665 1.644 1.671 1.659 1.213
As can be seen from the above table, in examples 1 to 4, the differences between the specific surface areas of the shell materials after pretreatment are not large, but the specific surface areas are significantly better than those in example 5, which indicates that tannic acid and diethyl oxalate have a gain effect on the increase of the specific surface area and the voids of the shell material.
B: pore size distribution curve
As shown in fig. 1, a distribution diagram of the adsorption pore diameter of the shell material before modification with chitosan prepared in example 3 shows that the average pore diameter of the shell material is 0.5-45 nm, wherein most of the pores with diameters smaller than 2nm are present, and the inner wall of the microporous material has good physical adsorption; the mesoporous material with the diameter of 2-50 nm is called as mesopore or mesopore, and has huge specific surface area and three-dimensional pore structure, and the two pore size distributions enable the material to have good adsorption effect and good slow release effect.
2) And (3) analyzing the shell powder material by a scanning electron microscope: the shell material before modification and the shell material before pretreatment of the chitosan prepared in example 3 were photographed at 1.00um by a Quanta200F field emission Scanning Electron Microscope (SEM).
As shown in attached figures 2 and 3, the pretreated shell powder material has more loose texture, larger pores are distributed on the surface, and the number and the size of the pores are obviously increased compared with those before pretreatment, which shows that the pretreatment has an effect of increasing the pores of the shell material.
Example 7:
adjusting test of water quality of eel breeding pond in net cage
In a certain eel breeding base, 4 breeding ponds with the same size are randomly selected, 20 net cages with the same size are placed in each pond, 100 eel fries with the same average size and weight are thrown into each net cage, and breeding is carried out under the same conditions, wherein the test period is 6 months. The results of the tests 1 to 3 were obtained by the methods of examples 3 to 5, and the results of the tests 2 were obtained by determining parameters at the end of the test period, using the method of adjusting the water quality without using the immobilized probiotic water quality control agent as the blank group.
TABLE 2 adjustment test results of water quality in pool for eel breeding in net cage
Figure BDA0001923775470000111
As can be seen from the above table, the difference between the total nitrogen content and the total phosphorus content in the aquaculture water bodies of the test groups 1 and 2 is not obvious, but is significantly lower than that of the test group 3; the test group 2 has the best effect of removing COD in the water body, and the difference of other test groups is not great; the difference in transparency between the test groups was significant, with test group 2 being the best; because the water quality is not regulated by using the immobilized probiotic water quality regulating agent during the blank group culture period, the data performance is the worst, the water quality is not good, the survival rate and the weight gain rate of the eels are influenced, and the culture benefit is not good easily; the test groups have excellent performance, particularly the survival rate of the test groups 1 and 2 reaches more than 90 percent, the weight gain rate is also obviously optimal, and the economic benefit is better.
Conventional techniques in the above embodiments are known to those skilled in the art, and therefore, will not be described in detail herein.
The above embodiments are merely illustrative, and not restrictive, and those skilled in the art can make various changes and modifications without departing from the spirit and scope of the invention. Therefore, all equivalent technical solutions also belong to the scope of the present invention, and the protection scope of the present invention should be defined by the claims.

Claims (7)

1. The immobilized probiotic water quality regulating agent is characterized in that: comprises functional shell material modified by chitosan and probiotics carried on the shell material; the specific surface area of the functional shell material before being modified by chitosan is 1.0-2.0 m2(ii)/g, the average pore diameter is 0.5-45 nm;
the functional shell material is subjected to calcination and water modification pretreatment before being modified by chitosan;
the pretreatment steps are as follows: cleaning shells, soaking the shells in 0.1-0.5% hydrochloric acid solution for 10-12 h, then placing the shells in a tubular furnace at 500-600 ℃ for anaerobic calcination for 1-2 h, taking out the shells and cooling the shells, crushing the shells and passing through a 100-120 mesh sieve, then adding 1-2% potassium carbonate solution with the weight of 5-10 times of the weight of the shells into the shells, ultrasonically dissolving the shells for 20-30 min under the power of 500-600W, then sending the mixed solution into a polytetrafluoroethylene reaction kettle, carrying out hydro-reforming reaction for 12-24 h at 150-180 ℃, centrifuging the solution, washing precipitates to be neutral, drying the precipitates for 6-8 h at 60-80 ℃, and obtaining the shells for later use;
in the step of pretreating the functional shell material, 0.15mM of tannic acid and 0.1mM of diethyl oxalate are contained in a potassium carbonate solution;
the functional shell material modified by chitosan is prepared by the following steps: adding the pretreated shell material into a chitosan solution, mixing and ultrasonically treating, and performing vacuum drying to obtain the chitosan-chitosan composite material; the chitosan solution contains kojic acid and sodium naphthenate;
the concentration of the chitosan solution is 1-2 mg/L, and the weight ratio of chitosan to the shell material is 1: 2-3 in terms of chitosan; the content of kojic acid and the content of sodium naphthenate in the chitosan solution are respectively 0.12-0.32 wt% and 0.15-0.5 wt%.
2. The immobilized probiotic water quality control agent according to claim 1, characterized in that: the immobilized probiotics is prepared by the following steps: adding 2-5 wt% of tea polyphenol into the activated and expanded probiotic culture solution, adding a chitosan modified functional shell material according to the proportion of 1: 0.5-1, magnetically stirring for 20-30 min, adding sodium alginate and 1: 0.3-0.5 of glycerol in the weight ratio of the sodium alginate to the shell material, uniformly mixing, standing for 1-2 h, and removing the upper clear water to obtain the probiotic culture solution.
3. The immobilized probiotic water quality control agent according to claim 1, characterized in that: the probiotic bacteria are selected from at least two of photosynthetic bacteria, bacillus subtilis, yeast, EM bacteria and lactic acid bacteria.
4. The method for adjusting the water quality of the eel breeding pond in the net cage is characterized in that: the adjusting method comprises the following steps:
a, periodically using the immobilized probiotic water quality regulator as defined in any one of claims 1 to 3;
b, stocking filter-feeding fishes and/or scrape-feeding fishes in the water outside the net cage;
and c, periodically transplanting and updating the submerged plants.
5. The method for adjusting the water quality of the eel breeding pond with the net cage according to claim 4, which is characterized in that: the use period of the immobilized probiotic water quality control agent is 1-2 months/time, and the use amount is 200-500 mg/m3A net cage.
6. The method for adjusting the water quality of the eel breeding pond with the net cage according to claim 4, which is characterized in that: the filter-feeding fishes are silver carps and bighead carps, and the scraper fishes are plagiognathops microlepis; the initial stocking density of the fishes is 10-20 tails/m3The average weight is 50-150 g.
7. The method for adjusting the water quality of the eel breeding pond with the net cage according to claim 4, which is characterized in that: the period for transplanting and updating the submerged plants is 45-60 days; the submerged plant is at least one selected from the group consisting of tape grass, hornwort, watermifoil and hydrilla.
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